Search Results (44)

Background/Objectives: Colorectal cancer (CRC) is the most common gastrointestinal (GI) malignancy, the second leading cause of cancer-related mortality, and the third most prevalent tumor. Around 20% of cases are metastatic or inoperable at diagnosis, often requiring palliative treatment, which may not be feasible in frail patients. Calcium-electroporation, a less invasive alternative, induces cell death via apoptosis, necrosis, and pyroptosis. This study is the first in the United Kingdom to evaluate the efficacy and safety of endoscopic calcium-electroporation in palliating distal CRC. Methods: Frail patients with inoperable left-sided CRC were included. The diagnosis and staging followed standard guidelines, while frailty was assessed using the performance status (PFS), Charlson comorbidity index (CCI), and American Society of Anesthesiologists (ASA) score. Calcium electroporation was performed via a flexible endoscopy usually under sedation, with symptom relief, quality of life (QoL), survival, and adverse events (AE) monitored. Results: Sixteen patients (median age 84.5) underwent 36 treatments with electroporation over 28 months (November 2022 to March 2025). The incidence of common symptoms was rectal bleeding (75%), constipation (25%), and pain (75%). Nine patients had metastases and three had failed conventional treatments. Symptomatic relief and an improved QoL occurred in 86.7%, with transfusion/iron infusion needs reduced by 91.7%. The median cancer-specific survival was 10 months, with a 94% survival rate. No device-related AE was recorded. One patient died after 11 months due to disease progression while two patients passed away from other medical conditions. Conclusions: Endoscopic calcium electroporation is a safe, palliative option effective for tumor reduction and symptomatic relief in frail CRC patients unfit for conventional therapies. Full article

RyR1-related myopathies (RyR1-RMs) include a wide range of genetic disorders that result from mutations in the RYR1 gene. Pathogenic variants lead to defective intracellular calcium homeostasis and muscle dysfunction. Fixing intracellular calcium leaks by stabilizing the RyR1 calcium channel has been identified as a promising therapeutic target. Gene therapy via prime editing also holds great promise as it can cure diseases by correcting genetic mutations. However, as more than 700 variants have been identified in the RYR1 gene, a universal treatment would be a more suitable solution for patients. Our investigation into the RyR1-S2843A mutation has yielded promising results. Using a calcium leak assay, we determined that the S2843A mutation was protective when combined with pathogenic mutations and significantly reduced the Ca²⁺ leak of the RyR1 channel. Our study demonstrated that prime editing can efficiently introduce the protective S2843A mutation. In vitro experiments using the RNA electroporation of the prime editing components in human myoblasts achieved a 31% introduction of this mutation. This article lays the foundation for a new therapeutic approach for RyR1-RM, where a unique once-in-a-lifetime prime editing treatment could potentially be universally applied to all patients with a leaky RyR1 channel. Full article

Electrochemotherapy (ECT) uses electroporation to enhance drug delivery into tumor cells, triggering bystander effects like immunogenicity and cell death. This study investigated bystander effects in vitro in 4T1 breast cancer cells following various electroporation treatments: reversible (1400 V/cm, 100 µs) bleomycin electrotransfer, irreversible (2800 V/cm, 100 µs) bleomycin electrotransfer, and calcium electroporation, including combinations. Conditioned media from treated cells (12–72 h incubation) were transferred to untreated cells, and viability was assessed via metabolic activity, cell count, and colony formation. A scratch assay evaluated wound healing. The bystander effect dramatically reduced colony formation, reaching 0% after bleomycin and calcium electrotransfer, and 2.37 ± 0.74% after irreversible electroporation (IRE). Metabolic activity decreased to 18.05 ± 6.77% and 11.62 ± 3.57% after bleomycin and calcium electrotransfer, respectively, and 56.21 ± 0.74% after IRE. Similarly, cell viability measured by flow cytometry was 10.00 ± 1.44%, 3.67 ± 0.32%, and 24.96 ± 1.37% after bleomycin electrotransfer, calcium electrotransfer, and IRE, respectively. Combined analysis of these effects yielded comparable results. Conditioned media, particularly from bleomycin electrotransfer and calcium electroporation, significantly reduced cell number, metabolic activity, and colony formation, demonstrating a strong bystander effect. Wound healing was also significantly delayed in groups exposed to conditioned media. Full article

The primary method of treatment for patients suffering from drug-resistant focal-onset epilepsy is resective surgery, which adversely impacts neurocognitive function. Radio frequency (RF) ablation and laser ablation are the methods with the most promise, achieving seizure-free rates similar to resection but with less negative impact on neurocognitive function. However, there remains a number of concerns and open technical questions about these two methods of thermal ablation, with the primary ones: (1) heating; (2) hemorrhage and bleeding; and (3) poor directionality. Irreversible electroporation (IRE) is a proven method of focal ablation, which circumvents all three of the primary concerns regarding focal RF and laser ablation. Here, we demonstrate the in vivo application of a flexible implant with organic electrodes for focal ablation of epilepsy foci using high-frequency IRE (H-FIRE) in mice. Our results show that local, targeted ablation is possible in the close neighborhood of the electrode, paving the way for the clinical application in the treatment of focal epilepsy. Full article

A healthy diet rich in plant proteins can help in preventing chronic degenerative diseases. Plant-based protein consists of derivatives from algae, fungi (like mushrooms) and other plant products including stems, leaves, fruits, vegetables, grains, seeds, legumes and nuts. These sources are not only rich in protein, but also contain a high percentage of iron, calcium, folates, fiber, carbohydrates, fats etc. Hence, it is essential to explore plant-based protein sources and their other nutritional components to address existing food insecurity issues. Nowadays, the impact of food processing has produced promising results in extracting valuable bio-compounds including proteins from the plant matrix. In this view, PEF technology has secured an exceptional place in solving food quality issues through minimized thermal effects in the samples, improved extraction capabilities at a shorter time, higher extraction levels, high nutritional content of extracted samples, greater shelf-life extension and increased microbial killing efficiency. It is an energy efficient process which is used as a pre-treatment to increase selective extraction of intracellular compounds through electroporation technique. Here, the processing parameters play a significant role in obtaining enhanced extraction levels. These parameters have also considerably influenced the protein digestibility and amino acid modification. So far, PEF has been producing remarkable results in plant protein extraction research. Among various plant sources mentioned above, there is a limited literature available on the use of PEF-assisted protein extraction from legumes. In this review, the authors have discussed essential legumes and their nutritional components and have highlighted how PEF can be beneficial in extracting the protein levels from these sources. Further research should focus on PEF-assisted protein extraction from legumes, specifically analyzing the properties of protein quality and quantity. Full article

Uveal melanoma (UM) represents a rare tumor of the uveal tract and is associated with a poor prognosis due to the high risk of metastasis. Despite advances in the treatment of UM, the mortality rate remains high, dictating an urgent need for novel therapeutic strategies. The current study introduces the first in vivo analysis of the therapeutic potential of calcium electroporation (CaEP) compared with electrochemotherapy (ECT) with bleomycin in a patient-derived xenograft (PDX) model based on the chorioallantoic membrane (CAM) assay. The experiments were conducted as monotherapy with either 5 or 10 mM calcium chloride or 1 or 2.5 µg/mL bleomycin in combination with EP or EP alone. CaEP and ECT induced a similar reduction in proliferative activity, neovascularization, and melanocytic expansion. A dose-dependent effect of CaEP triggered a significant induction of necrosis, whereas ECT application of 1 µg/mL bleomycin resulted in a significantly increased apoptotic response compared with untreated tumor grafts. Our results outline the prospective use of CaEP and ECT with bleomycin as an adjuvant treatment of UM, facilitating adequate local tumor control and potentially an improvement in metastatic and overall survival rates. Full article

Electroporation with chemotherapy (ECT) is currently offered as a treatment in Europe for locoregional or metastatic melanoma with cutaneous lesions. However, the role of surgery and other forms of electroporation in melanoma requires further evaluation. Two reviewers used two databases to conduct a literature search and review, and 51 publications related to electroporation with chemotherapy, immunotherapy, or gene delivery were found. ECT appears to be effective in reducing tumor burden for surgical resection, replacing surgical intervention with evidence of complete regression in some lesions, and inducing both local and systemic immune effects. These immune effects are pronounced when ECT is combined with immunotherapy, with a statistically significant improvement in overall survival (OS). Other forms of electroporation, such as those using calcium chloride, an IL-12 plasmid, and vaccination, require further study. However, IL-12 plasmid electroporation may be inferior to ECT based on the evidence available. Furthermore, irradiation of the tumor prior to ECT treatment is negatively correlated with local response. Access to ECT is restricted in the US and requires further evaluation. More randomized controlled trials of ECT and electroporation treatment in locoregional melanoma are recommended. Full article

Pulsed field ablation (PFA) is a promising new treatment for atrial fibrillation (AF), in which pulmonary vein isolation is achieved by irreversible electroporation. Electroporation causes ATP to leak through the permeabilized membrane. ATP is required both for the healing of the cell membrane and for the functioning of ion pumps, such as sarco/endoplasmic reticulum Ca²⁺-ATPase (SERCA) or Na⁺,K⁺-ATPase (NKA), which play a key role in maintaining continuous contractions of the heart muscle. We investigated the effects of electroporation on the expression of ion pumps and possible correlations with the activation of AMPK, the main energy sensor in cells. H9c2 rat cardiac cells were exposed to either monopolar or bipolar (H-FIRE) pulses. Cells lysed 4 or 24 h after electroporation were used for mRNA and protein expression analyses. Overall, both pulse protocols caused a dose-dependent downregulation of crucial SERCA and NKA isoforms, except for NKAα2 and β3, which were upregulated after 24 h. Monopolar pulses also decreased the phosphorylation of FXYD1, which may cause an inhibition of NKA activity. Both pulse protocols caused an increased AMPK activity, which may decrease both SERCA and NKA activity via calcium/calmodulin-dependent protein kinase. Our results provide important new insights into what happens in surviving cardiomyocytes after they are exposed to PFA. Full article

Despite recent advancements in the diagnosis and treatment of uveal melanoma (UM), its metastatic rate remains high and is accompanied by a highly dismal prognosis, constituting an unmet need for the development of novel adjuvant therapeutic strategies. We established an in vivo chick chorioallantoic membrane (CAM)-based UM xenograft model from UPMD2 and UPMM3 cell lines to examine its feasibility for the improvement of selection of drug candidates. The efficacy of calcium electroporation (CaEP) with 5 or 10 mM calcium chloride (Ca) and electrochemotherapy (ECT) with 1 or 2.5 µg/mL bleomycin in comparison to monotherapy with the tested drug or electroporation (EP) alone was investigated on the generated UM tumors. CaEP and ECT showed a similar reduction of proliferation and melanocytic expansion with a dose-dependent effect for bleomycin, whereas CaEP induced a significant increase of the apoptosis and a reduction of vascularization with varying sensitivity for the two xenograft types. Our in vivo results suggest that CaEP and ECT may facilitate the adequate local tumor control and contribute to the preservation of the bulbus, potentially opening new horizons in the adjuvant treatment of advanced UM. Full article

In recent years, the application of pulsed electric fields with very short durations (nanoseconds) and extremely high amplitudes (MV/m) has been investigated for novel medical purposes. Various electric protocols have been explored for different objectives, including the utilization of fractionated pulse doses to enhance cell electrosensitization to the uptake of different markers or an increase in apoptosis. This study focused on the use of fluorescence imaging to examine molecular calcium fluxes induced by different fractionated protocols of short electric pulses in neuroblastoma (SH-SY5Y) and mesenchymal stem cells (HaMSCs) that were electroporated using nanosecond pulsed electric fields. In our experimental setup, we did not observe cell electrosensitization in terms of an increase in calcium flux following the administration of fractionated doses of nanosecond pulsed electric fields with respect to the non-fractionated dose. However, we observed the targeted activation of calcium-dependent genes (c-FOS, c-JUN, EGR1, NURR-1, β3-TUBULIN) based on the duration of calcium flux, independent of the instantaneous levels achieved but solely dependent on the final plateau reached. This level of control may have potential applications in various medical and biological treatments that rely on calcium and the delivery of nanosecond pulsed electric fields. Full article

Cheese powder is a multifunctional ingredient that is produced by spray drying a hot cheese emulsion called cheese feed. Feed stability is achieved by manipulating calcium equilibrium using emulsifying salts. However, the increased demand for ‘green’ products created a need for alternative production methods. Therefore, this study investigated the impact of ohmic heating (OH) on Cheddar cheese, mineral balance, and the resulting cheese feed characteristics compared with a conventional method. A full factorial design was implemented to determine the optimal OH parameters for calcium solubilization. Electric field exposure and temperature had a positive correlation with mineral solubilization, where temperature had the greatest impact. Structural differences in pre-treated cheeses (TC) were analyzed using thermorheological and microscopic techniques. Obtained feeds were analyzed for particle size, stability, and viscosity. OH-treatment caused a weaker cheese structure, indicating the potential removal of calcium phosphate complexes. Lower component retention of OH_TC was attributed to the electroporation effect of OH treatment. Microscopic images revealed structural changes, with OH_TC displaying a more porous structure. Depending on the pre-treatment method, component recovery, viscosity, particle size distribution, and colloidal stability of the obtained feeds showed differences. Our findings show the potential of OH in mineral solubilization; however, further improvements are needed for industrial application. Full article

Calcium electroporation (CaEP) is an innovative approach to treating cancer, involving the internalization of supraphysiological amounts of calcium through electroporation, which leads to cell death. CaEP enables the replacement of chemotherapeutics (e.g., bleomycin). Here, we present a standard microsecond (μsCaEP) and novel high-frequency nanosecond protocols for calcium electroporation (nsCaEP) for the elimination of carcinoma tumors in C57BL/6J mice. We show the efficacy of CaEP in eliminating tumors and increasing their survival rates in vivo. The antitumor immune response after the treatment was observed by investigating immune cell populations in tumors, spleens, lymph nodes, and blood, as well as assessing antitumor antibodies. CaEP treatment resulted in an increased percentage of CD4⁺ and CD8⁺ central memory T cells and decreased splenic myeloid-derived suppressor cells (MDSC). Moreover, increased levels of antitumor IgG antibodies after CaEP treatment were detected. The experimental results demonstrated that the administration of CaEP led to tumor growth delay, increased survival rates, and stimulated immune response, indicating a potential synergistic relationship between CaEP and immunotherapy. Full article

ATP, as a paracrine signalling molecule, induces intracellular Ca²⁺ elevation via the activation of purinergic receptors on the surface of glia-like cochlear supporting cells. These cells, including the Deiters’ cells (DCs), are also coupled by gap junctions that allow the propagation of intercellular Ca²⁺ waves via diffusion of Ca²⁺ mobilising second messenger IP₃ between neighbouring cells. We have compared the ATP-evoked Ca²⁺ transients and the effect of two different gap junction (GJ) blockers (octanol and carbenoxolone, CBX) on the Ca²⁺ transients in DCs located in the apical and middle turns of the hemicochlea preparation of BALB/c mice (P14–19). Octanol had no effect on Ca²⁺ signalling, while CBX inhibited the ATP response, more prominently in the middle turn. Based on astrocyte models and using our experimental results, we successfully simulated the Ca²⁺ dynamics in DCs in different cochlear regions. The mathematical model reliably described the Ca²⁺ transients in the DCs and suggested that the tonotopical differences could originate from differences in purinoceptor and Ca²⁺ pump expressions and in IP₃–Ca²⁺ release mechanisms. The cochlear turn-dependent effect of CBX might be the result of the differing connexin isoform composition of GJs along the tonotopic axis. The contribution of IP₃-mediated Ca²⁺ signalling inhibition by CBX cannot be excluded. Full article

The 4th World Congress of Electroporation (Copenhagen, 9–13 October 2022) provided a unique opportunity to convene leading experts in pulsed electric fields (PEF). PEF-based therapies harness electric fields to produce therapeutically useful effects on cancers and represent a valuable option for a variety of patients. As such, irreversible electroporation (IRE), gene electrotransfer (GET), electrochemotherapy (ECT), calcium electroporation (Ca-EP), and tumour-treating fields (TTF) are on the rise. Still, their full therapeutic potential remains underappreciated, and the field faces fragmentation, as shown by parallel maturation and differences in the stages of development and regulatory approval worldwide. This narrative review provides a glimpse of PEF-based techniques, including key mechanisms, clinical indications, and advances in therapy; finally, it offers insights into current research directions. By highlighting a common ground, the authors aim to break silos, strengthen cross-functional collaboration, and pave the way to novel possibilities for intervention. Intriguingly, beyond their peculiar mechanism of action, PEF-based therapies share technical interconnections and multifaceted biological effects (e.g., vascular, immunological) worth exploiting in combinatorial strategies. Full article

Breast cancer ranks among the top three most common malignant neoplasms in Poland. The use of calcium ion-assisted electroporation is an alternative approach to the classic treatment of this disease. The studies conducted in recent years confirm the effectiveness of electroporation with calcium ions. Electroporation is a method that uses short electrical pulses to create transitional pores in the cell membrane to allow the penetration of certain drugs. The aim of this study was to investigate the antitumor effects of electroporation alone and calcium ion-assisted electroporation on human mammary adenocarcinoma cells that are sensitive (MCF-7/WT) and resistant to doxorubicin (MCF-7/DOX). The cell viability was assessed using independent tests: MTT and SRB. The type of cell death after the applied therapy was determined by TUNEL and flow cytometry (FACS) methods. The expression of Cav3.1 and Cav3.2 proteins of T-type voltage-gated calcium channels was assessed by immunocytochemistry, and changes in the morphology of CaEP-treated cells were visualized using a holotomographic microscope. The obtained results confirmed the effectiveness of the investigated therapeutic method. The results of the work constitute a good basis for planning research at the in vivo level and in the future to develop a more effective and safer method of breast cancer treatment for patients. Full article