Epigenetic regulation can play a dual role in viral infection, acting as part of the host defense, while being hijacked to control viral latency, replication, and persistence. During the COVID-19 pandemic, substantial evidence indicated that different SARS-CoV-2 variants may lead to distinct outcomes by affecting different molecular pathways, including epigenetic mechanisms. In this study, we assessed miRNA expression using high-throughput microarray technology to simultaneously measure the expression of thousands of miRNAs. We focused on: comparing Delta and Omicron infections with healthy controls; identifying specific miRNAs linked to symptom severity; analyzing miRNA expression across different stages of Delta variant infection; and predicting miRNA target genes and evaluating miRNA–mRNA interactions. We identified 65 miRNAs that were significantly differentially expressed simultaneously in both SARS-CoV-2 variants compared to healthy samples. These miRNAs were categorized into distinct functional groups based on their roles in viral infection. We observed an association between the expression of specific miRNAs and virus-induced symptom severity and patients’ outcomes in both variants. Furthermore, we examined miRNA expression across the various phases of the Delta variant and identified 38 miRNAs that were significantly differentially expressed. Finally, mRNA-miRNA interaction analysis revealed PTEN, IGF1R, MYC, and STAT3 as the most interacting genes. Profiling thousands of miRNAs in response to different SARS-CoV-2 variants offers new opportunities for diagnosis, prognosis, and therapy. Although comorbidities and pathway cross-talk may influence the obtained results, our findings provide novel insights into the host response to SARS-CoV-2 infection at the epigenetic level.

Breast cancer remains one of the most diagnosed and lethal cancers, with increased incidence in developing countries. Empagliflozin, a sodium-glucose cotransporter 2 (SGLT2) inhibitor, is an effective and well-tolerated antidiabetic drug for managing and treating type 2 diabetes mellitus. Moreover, it can reduce the risk of death from cardiovascular causes and hospitalizations for heart failure. Recent data indicate that empagliflozin has antioxidant and anti-inflammatory effects mediated by modulating various signaling pathways. Molecular and cellular analysis revealed that SGLT2 is expressed in different types of cancer, including breast, cervical, and lung. We analyzed the potential anticancer activities of empagliflozin in the 4T1 breast cancer cell line. To assess the effect of the SGLT2 inhibitor on the migration and clonogenicity of cancer cells, we used the wound healing assay and the colony formation assay. The acquired data showed some trends; however, no significant results were obtained.

Background:Cardiovascular diseases (CVD) remain the leading cause of premature death worldwide, with a steadily rising incidence. Ischemia, characterized by insufficient blood flow and subsequent tissue hypoxia, contributes significantly to cellular damage, dysfunction, as well as epigenetic alterations in CVD. Hypoxia-induced epigenetic changes have been associated with fibrosis, inflammation, metabolic dysregulation, and altered programmed cell death in cardiac tissue. Empagliflozin, a sodium-glucose cotransporter 2 (SGLT2) inhibitor primarily prescribed for type 2 diabetes, has also demonstrated beneficial effects in patients with CVD. Methods:The present study aimed to evaluate the protective role of Empagliflozin in primary human cardiomyocytes under chemically induced hypoxia, with a focus on its influence on specific microRNAs. Cardioprotective effects were analyzed through the expression of miR-214-3p, miR-22-5p, miR-103-5p, and miR-145-5p, along with protein levels of GAPDH and HIF1α. Results and conclusion:After 24 h of Empagliflozin treatment, a significant upregulation of the cardioprotective miR-214-3p (p < 0.05) was observed. These findings demonstrate a direct cardioprotective effect of Empagliflozin in cobalt-treated cardiomyocytes and suggest its potential role as an epigenetic modulator in the pathogenesis of CVD.

Anew in vitromodel for modeling Duchenne muscular dystrophy, derived from pediatric patient with a unique mutation in DMD gene is introduced. Peripheral blood was used as asource for primary cells (mononuclear cells) and subsequently reprogrammed into induced pluripotent stem cells with synthetic Sendai vector. Reprogrammed iPS cells showed the expression of pluripotency factors and the ability to differentiateinto all three germ layers.

Colorectal cancer (CRC) is a major global health problem, with type 2 diabetes mellitus (T2DM) and obesity among its significant risk factors. Empagliflozin (EMPA), a selective sodium–glucose cotransporter 2 (SGLT2) inhibitor widely used in the treatment of T2DM, has also demonstrated potential anticancer effects. However, reports of possible adverse effects in certain oncological contexts have raised concerns. We established several in vitro assays to evaluate the effects of EMPA on different colon cancer–derived cell lines. We applied proliferation, wound-healing, colony-formation, and CCK-8 assays, together with measurements of EMPA and glucose consumption, in HCT116, HT29, DLD1, SW480, and LS180 CRC cell lines. SGLT2 expression was confirmed in all cell lines. EMPA reduced proliferation in HT29 cells. Clonogenicity was decreased in HT29 and HCT116, but increased in DLD1 and SW480. Migration was reduced in DLD1, whereas enhanced migratory potential was observed in HT29 and HCT116. Cell viability was not significantly affected at the tested concentrations. HT29 cells showed significantly higher EMPA consumption compared with other lines. Glucose consumption differed among cell lines but was not altered by EMPA treatment. LS180 cells showed no significant response in the assessed parameters. EMPA was associated with ambivalent, cell line-specific effects on CRC cell behavior. While antitumor activity was observed in some models, enhanced migration and clonogenicity were detected in others. These findings warrant further investigation regarding the use of EMPA in patients with T2DM and concomitant CRC.

Wastewater-based epidemiology (WBE) represents a promising method for early detection and ongoing monitoring of various pathogens. This epidemiological approach has recently attracted significant research interest. During the SARS-CoV-2 pandemic, WBE was estimated as a complementary surveillance tool for mapping the spread of the virus within the population. Although this method has great potential for pathogen monitoring at both local and national levels, there are some limitations, notably the presence of high concentrations of inhibitors that contaminate extracted nucleic acids from wastewater, which can lead to false negative results. This proof-of-concept study focuses on detecting RNA and DNA viruses, specifically SARS-CoV-2 and adenovirus, in wastewater by droplet digital PCR (ddPCR) and compares the results with those obtained with the routinely used RT-qPCR and qPCR methods. The results of our proof-of-concept study indicate that WBE represents a promising approach for monitoring pathogens such as SARS-CoV-2 and adenovirus; however, challenges such as contaminants in wastewater need to be addressed to increase detection accuracy. Due to this limitation, novel technologies such as ddPCR offer new opportunities to improve epidemiological monitoring of various infectious agents in the population.

The coronavirus disease 2019 (COVID-19) has highly impacted everyday lives and healthcare systems since its spread across the globe. As a result of numerous mutations in the viral genome, several genetic variants and sub-variants have evolved, and they differ in various characteristics, such as infectivity, transmissibility, and immune response in human body. The aim of this study was to compare cytokine levels in plasma samples from patients with acute COVID-19, infected with either the Delta or Omicron variants. This retrospective study included 80 hospitalized patients with PCR-confirmed SARS-CoV-2. Blood samples from the patients as well as control subjects were collected, and the multiplex cytokine assay was performed determining the levels of 48 cytokines in plasma. This study included 50 hospitalized patients with Delta, 30 patients with Omicron variant, and 18 control subjects. Severe and critical disease course led to 27 significantly modified levels of cytokines in both COVID-19 variants compared to control subjects. However, 12 cytokine levels (GRO-α, IFN-α2, IL-1α, IL-4, IL-9, IL-16, MIF, MIP-1β, SDF-1α, TNF-α, TNF-β, TRAIL) were significantly elevated in Delta versus Omicron variant with the discrimination equal to AUC = 0.876. Moreover, vaccination significantly decreased levels of GM-CSF, IL-7, and IL-8 in patients with Omicron variant. This study suggests that the genetic variants of SARS-CoV-2 affect the immune response differently.

Cardiovascular diseases (CVD) are the leading cause of premature death and disability. Hypoxic conditions play a central role in the pathophysiology of all CVD. Empagliflozin (EMPA), a sodium-glucose cotransporter 2 (SGLT2) inhibitor used for diabetes mellitus type II therapy, has demonstrated a beneficial role in improving cardiovascular outcomes for patients with heart failure. Our study aimed to assess the cardioprotective effect of EMPA on primary human cardiomyocytes in a chemically induced hypoxia model. The cardioprotective effect of the SGLT2 inhibitor was evaluated through four individual experiments including: (1) evaluating mitochondrial network integrity, (2) determining cell count, (3) metabolomic profiling, and (4) determining alterations in miRNA expression. After 24 h of EMPA treatment, we observed a significant improvement in mitochondrial network complexity, as evidenced by increased branching (p < 0.05) and a reduced number of rod-shaped mitochondria (p < 0.05) in EMPA-treated cells compared to controls. After cobalt treatment, we didn't observe any protective effect of EMPA in cells affected by cobalt in various biological aspects, including miRNA expression, metabolomics, or viability. Although EMPA treatment was not able to propagate beneficial effects in the presence of cobalt, pretreatment of cells with EMPA indicated a potential cardioprotective effect associated with improving mitochondrial morphology.

BackgroundThe differential diagnosis of lymphadenopathy is an important determinant of prognosis in patients with breast cancer (BC). Invasive, fine needle aspiration (FNA) biopsy has been long considered as the gold standard for differentiating malignant lymph nodes (LN) from benign ones. Ultrasonography (USG) evaluation is a useful, rapid, and user-friendly imaging tool for LN assessment due to its high resolution. Compared to USG, ultrasound elastography is a relatively novel non-invasive method to differentiate benign and malignant lesions based on the stiffness heterogeneity of the tissue. The purpose of our study was to compare non-invasive imaging techniques, conventional USG, and strain elastography, to differentiate benign and malignant LNs lesions in a cohort of patients with early BC.MethodsIn total, 50 patients (48 women and 2 men) with histologically confirmed early BC were evaluated by conventional USG in B-mode followed by strain elastography (using parameters: pattern, strain ratio, hue histogram) for assessment of axillary LNs status. The surgical treatment included surgery of regional LNs (sentinel LN biopsy or axillary dissection), which served as the gold standard in statistical processing.ResultsThe USG B-mode was found to have a sensitivity of 68.75% and a specificity of 61.54%. Among strain elastography parameters, the elastographic pattern showed the highest specificity (66.67%) while the sensitivity was 83.3%. The strain ratio showed 100% sensitivity and 55.6% specificity, followed by a hue histogram with a sensitivity of 72.2%, but specificity was only 25.9%.ConclusionDespite promising data, monitored parameters currently cannot reliably replace sentinel LN biopsy. However, the monitored parameters represent an appropriate additional tool that can be used to refine preoperative staging, better targeting of FNA biopsy, and more accurate assessment of LNs in follow-up patients within the dispensary.

Purpose: Uveal melanoma (UM) is the most prevalent primary intraocular malignancy in adults, with prognosis significantly influenced by genetic and epigenetic factors. Reliable and cost-effective methods to detect chromosomal aberrations and DNA methylation changes are essential for improving prognostication and informing treatment strategies in UM. This study evaluated the effectiveness of multiplex ligation-dependent probe amplification (MLPA) in detecting UM-specific copy number variations (CNVs) and promoter methylation changes across 25 tumor suppressor genes (TSGs). Methods: DNA from 58 UM tissues was analyzed with the SALSA MLPA Probemix P027 Uveal melanoma kit, and a subset of 18 samples was further assessed using the SALSA MLPA Probemix ME002-C1 Tumour suppressor mix 2 kit to identify key CNVs and methylation alterations linked to poor prognosis. Validation was carried out with a high-resolution comparative genomic hybridization (CGH) array on 10 samples and the Illumina Infinium Methylation EPIC v1.0 BeadChip array on 25 samples. Results: Our findings indicate that MLPA is a versatile and robust method for detecting CNVs, showing strong correlations with CGH data and highlighting specific CNV patterns linked to clinical outcomes in UM. However, the ME002-C1 kit showed limited utility for comprehensive methylation analysis, as differential methylation was not observed in the studied TSG loci. Conclusions: Although MLPA effectively identifies CNVs relevant to UM prognosis, integrating additional methylation-specific approaches could broaden the scope of DNA methylation analysis, offering a more comprehensive molecular understanding of UM that may enhance prognostication and personalized treatment.

The choice of appropriate analytical methods for determining methylation patterns at specific loci across the genome is essential for identifying novel diagnostic and prognostic markers for subsequent clinical implementation. Various methods exist for determining methylation status using different technologies. In this study, we compared two distinct digital polymerase chain reaction (PCR) platforms: the nanoplate-based Qiagen QIAcuity Digital PCR (dPCR) System and the droplet-based Bio-Rad QX-200 Droplet Digital PCR (ddPCR) System. By assessing their efficacy and other attributes, we aimed to elucidate each platform's strengths and limitations in the sensitive detection of DNA methylation, thus contributing valuable insights to the field of molecular diagnostics. We analyzed the methylation status of the CDH13 gene in 141 formalin-fixed, paraffin-embedded breast cancer tissue samples using our in-house developed methylation-specific labeled assay. The specificity and sensitivity of the CDH13 assay evaluated by dPCR were 99.62% and 99.08%, respectively; ddPCR analysis reached a specificity of 100% and a sensitivity of 98.03%. In addition, our data revealed a strong correlation between the methylation levels measured by both methods (r = 0.954). Although both methods are based on different technologies, they yielded comparable, highly sensitive experimental data in our study. Consequently, the main criteria for selecting an optimal digital PCR platform for methylation analysis may lie in other factors such as workflow time and complexity, instrument requirements, the possibility of temperature gradient, reanalysis, or offline options.

Background: Bronchial asthma, the most prevalent chronic inflammatory airway disease in children, exhibits a concerning rise in both incidence and prevalence. Asthma biomarkers hold promise for stratifying patients into distinct clinical phenotypes, paving the way for targeted and personalized treatment approaches. Aim of Study: This study aimed to evaluate the association between novel and non-established semi-invasive circulating and wellknown exhaled inflammatory biomarkers in two distinct pediatric asthma populations stratified by disease severity. Materials and Methods: Forty-four asthmatic children aged 8-12 years meeting inclusion criteria were recruited from hospitalized patients. The first group (n=15, mean age 9.8 years) consisted of patients with mild persistent asthma who did not require regular inhaled corticosteroids (ICS). The second group (n=29, mean age 9.8 years) consisted of children with moderate to persistent asthma who received regular ICS treatment. Serum levels of interleukins (IL-13, IL-1(3), eosinophil-derived neurotoxin (EDN), and surfactant protein D (SPD) were measured by ELISA in all participants. In addition, exhaled nitric oxide (FeNO) and blood eosinophil counts were evaluated. Results: No significant differences were observed in the baseline plasma concentrations of inflammatory markers (IL-13, IL-1(3, SPD, and EDN) or exhaled FeNO between the ICS-treated and non-ICS-treated groups. Further inter-individual analysis confirmed significant positive correlations between IL-13, SPD, and IL-1(3 (Pearson's r = 0.591-0.781) in both groups of patients. Interestingly, the ICS-treated group compared to the nontreated group showed an exclusive moderate negative correlation between FeNO and IL-1(3. In contrast, FeNO exhibited a positive correlation with EDN and a strong association with eosinophil count in all the study groups. Conclusion: Our findings highlight the complex and unresolved role of asthma biomarkers in routine clinical practice for the management of childhood asthma, particularly in predicting exacerbations. By comparing the relationships of carefully selected biomarkers, we can achieve a greater clinical predictive value.

Colorectal cancer (CRC) is one of the leading oncogenic disorders, both in terms of incidence and mortality. The etiology of the disease is certainly multifactorial. Various risk factors like alcohol consumption, smoking, CRC family history, inflammatory bowel disease, hormone therapy, aspirin/nonsteroidal anti-inflammatory drugs use, higher body mass index, consumption of red and/or processed meat, insufficient physical activity, and decreased intake of fruit and vegetables have been pointed out; however, there is not enough support evidence for a single particular causative mechanism. Recently, gut bacterial microbiota has been shown to influence significantly the pathogenesis of CRC. However, little attention is paid to the putative impact of plasmids in gut flora.

Background: Cancer management faces multiple obstacles, including resistance to current therapeutic approaches. In the face of challenging microenvironments, cancer cells adapt metabolically to maintain their supply of energy and precursor molecules for biosynthesis and thus sustain rapid proliferation and tumor growth. Among the various metabolic adaptations observed in cancer cells, the altered glucose metabolism is the most widely studied. The aberrant glycolytic modification in cancer cells has been associated with rapid cell division, tumor growth, cancer progression, and drug resistance. The higher rates of glycolysis in cancer cells, as a hallmark of cancer progression, is modulated by the transcription factor hypoxia inducible factor 1 alpha (HIF-1a), a downstream target of the PI3K/Akt signaling, the most deregulated pathway in cancer.Aim of Review: We provide a detailed overview of current, primarily experimental, evidence on the potential effectiveness of flavonoids to combat aberrant glycolysis-induced resistance of cancer cells to conventional and targeted therapies. The manuscript focuses primarily on flavonoids reducing cancer resistance via affecting PI3K/Akt, HIF-1a (as the transcription factor critical for glucose metabolism of cancer cells that is regulated by PI3K/Akt pathway), and key glycolytic mediators downstream of PI3K/Akt/HIF-1a signaling (glucose transporters and key glycolytic enzymes). Key ScientificConcepts of Review: The working hypothesis of the manuscript proposes HIF-1a - the transcription factor critical for glucose metabolism of cancer cells regulated by PI3K/Akt pathway as an attractive target for application of flavonoids to mitigate cancer resistance. Phytochemicals represent a source of promising substances for cancer management applicable to primary, secondary, and tertiary care. However, accurate patient stratification and individualized patient profiling represent crucial steps in the paradigm shift from reactive to predictive, preventive, and personalized medicine (PPPM / 3PM). The article is focused on targeting molecular patterns by natural substances and provides evidence based recommendations for the 3PM relevant implementation.(c) 2023 The Authors. Published by Elsevier B.V. on behalf of Cairo University. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).

Introduction: Based on extensive data from oncology research, the use of phytochemicals or plant-based nutraceuticals is considered an innovative tool for cancer management. This research aimed to analyze the oncostatic properties of Salvia officinalis L. [Lamiaceae; Salviae officinalis herba] using animal and in vitro models of breast carcinoma (BC).Methods: The effects of dietary administered S. officinalis in two concentrations (0.1%/SAL 0.1/and 1%/SAL 1/) were assessed in both syngeneic 4T1 mouse and chemically induced rat models of BC. The histopathological and molecular evaluations of rodent carcinoma specimens were performed after the autopsy. Besides, numerous in vitro analyses using two human cancer cell lines were performed.Results and Conclusion: The dominant metabolites found in S. officinalis propylene glycol extract (SPGE) were representatives of phenolics, specifically rosmarinic, protocatechuic, and salicylic acids. Furthermore, the occurrence of triterpenoids ursolic and oleanolic acid was proved in SPGE. In a mouse model, a non-significant tumor volume decrease after S. officinalis treatment was associated with a significant reduction in the mitotic activity index of 4T1 tumors by 37.5% (SAL 0.1) and 31.5% (SAL 1) vs. controls (set as a blank group with not applied salvia in the diet). In addition, salvia at higher doses significantly decreased necrosis/whole tumor area ratio by 46% when compared to control tumor samples. In a rat chemoprevention study, S. officinalis at a higher dose significantly lengthened the latency of tumors by 8.5 days and significantly improved the high/low-grade carcinomas ratio vs. controls in both doses. Analyses of the mechanisms of anticancer activities of S. officinalis included well-validated prognostic, predictive, and diagnostic biomarkers that are applied in both oncology practice and preclinical investigation. Our assessment in vivo revealed numerous significant changes after a comparison of treated vs. untreated cancer cells. In this regard, we found an overexpression in caspase-3, an increased Bax/Bcl-2 ratio, and a decrease in MDA, ALDH1, and EpCam expression. In addition, salvia reduced TGF-β serum levels in rats (decrease in IL-6 and TNF-α levels were with borderline significance). Evaluation of epigenetic modifications in rat cancer specimens in vivo revealed a decline in the lysine methylations of H3K4m3 and an increase in lysine acetylation in H4K16ac levels in treated groups. Salvia decreased the relative levels of oncogenic miR21 and tumor-suppressive miR145 (miR210, miR22, miR34a, and miR155 were not significantly altered). The methylation of ATM and PTEN promoters was decreased after S. officinalis treatment (PITX2, RASSF1, and TIMP3 promoters were not altered). Analyzing plasma metabolomics profile in tumor-bearing rats, we found reduced levels of ketoacids derived from BCAAs after salvia treatment. In vitro analyses revealed significant anti-cancer effects of SPGE extract in MCF‐7 and MDA-MB-231 cell lines (cytotoxicity, caspase‐3/-7, Bcl‐2, Annexin V/PI, cell cycle, BrdU, and mitochondrial membrane potential). Our study demonstrates the significant chemopreventive and treatment effects of salvia haulm using animal or in vitro BC models.

Identifying novel epigenetic biomarkers is a promising way to improve the clinical management of patients with breast cancer. Our study aimed to determine the methylation pattern of 25 tumor suppressor genes (TSG) and select the best methylation biomarker associated with clinicopathological features in the cohort of Slovak patients diagnosed with invasive ductal carcinoma (IDC). Overall, 166 formalin-fixed, paraffin-embedded (FFPE) tissues obtained from patients with IDC were included in the study. The methylation status of the promoter regions of 25 TSG was analyzed using semiquantitative methylation-specific MLPA (MS-MLPA). We identified CDH13 as the most frequently methylated gene in our cohort of patients. Further analysis by ddPCR confirmed an increased level of methylation in the promoter region of CDH13. A significant difference in CDH13 methylation levels was observed between IDC molecular subtypes LUM A versus HER2 (P = 0.0116) and HER2 versus TNBC (P = 0.0234). In addition, significantly higher methylation was detected in HER2+ versus HER2- tumors (P = 0.0004) and PR− versus PR+ tumors (P = 0.0421). Our results provide evidence that alteration in CDH13 methylation is associated with clinicopathological features in the cohort of Slovak patients with IDC. In addition, using ddPCR as a methylation-sensitive method represents a promising approach characterized by higher precision and technical simplicity to measure the methylation of target CpGs in CDH13 compared to other conventional methods such as MS-MLPA.

Cancer development and progression are characterized by intricate genetic and epigenetic alternations, with DNA methylation playing a crucial role in this transformative process. Recent advancements in DNA methylation mapping technologies have significantly contributed to the identification of novel cancer biomarkers. We provide a short overview of Conformité Européenne-marked in vitro diagnostic tests designed to detect DNA methylation changes in oncology practice.

IntroductionWithin oncology research, there is a high effort for new approaches to prevent and treat cancer as a life-threatening disease. Specific plant species that adapt to harsh conditions may possess unique properties that may be utilized in the management of cancer.HypothesisChokeberry fruit is rich in secondary metabolites with anti-cancer activities potentially useful in cancer prevention and treatment.Aims of the study and MethodsBased on mentioned hypothesis, the main goal of our study was to evaluate the antitumor effects of dietary administered Aronia melanocarpa L. fruit peels (in two concentrations of 0.3 and 3% [w/w]) in the therapeutic syngeneic 4T1 mouse adenocarcinoma model, the chemopreventive model of chemically induced mammary carcinogenesis in rats, a cell antioxidant assay, and robust in vitro analyses using MCF-7 and MDA-MB-231 cancer cells.ResultsThe dominant metabolites in the A. melanocarpa fruit peel extract tested were phenolic derivatives classified as anthocyanins and procyanidins. In a therapeutic model, aronia significantly reduced the volume of 4T1 tumors at both higher and lower doses. In the same tumors, we noted a significant dose-dependent decrease in the mitotic activity index compared to the control. In the chemopreventive model, the expression of Bax was significantly increased by aronia at both doses. Additionally, aronia decreased Bcl-2 and VEGF levels, increasing the Bax/Bcl-2 ratio compared to the control group. The cytoplasmic expression of caspase-3 was significantly enhanced when aronia was administered at a higher dosage, in contrast to both the control group and the aronia group treated with a lower dosage. Furthermore, the higher dosage of aronia exhibited a significant reduction in the expression of the tumor stem cell marker CD133 compared to the control group. In addition, the examination of aronia`s epigenetic impact on tumor tissue through in vivo analyses revealed significant alterations in histone chemical modifications, specifically H3K4m3 and H3K9m3, miRNAs expression (miR155, miR210, and miR34a) and methylation status of tumor suppressor genes (PTEN and TIMP3). In vitro studies utilizing a methanolic extract of A.melanocarpa demonstrated significant anti-cancer properties in the MCF-7 and MDA-MB-231 cell lines. Various analyses, including Resazurin, cell cycle, annexin V/PI, caspase-3/7, Bcl-2, PARP, and mitochondrial membrane potential, were conducted in this regard. Additionally, the aronia extract enhanced the responsiveness to epirubicin in both cancer cell lines.ConclusionThis study is the first to analyze the antitumor effect of A. melanocarpa in selected models of experimental breast carcinoma in vivo and in vitro. The utilization of the antitumor effects of aronia in clinical practice is still minimal and requires precise and long-term clinical evaluations. Individualized cancer-type profiling and patient stratification are crucial for effectively implementing plant nutraceuticals within targeted anti-cancer strategies in clinical oncology.

DNA methylation is characterized as a gene regulatory mechanism that involves the methylation of the 5-carbon (C5) position of cytosine, resulting in the formation of 5-methylcytosine. The analysis of aberrantly methylated cytosine-phosphate-guanine (CpG) dinucleotides, primarily in the promoter regions of tumor suppressor genes, can serve as promising prognostic and predictive markers of cancer development. Meis homeobox 1 (MEIS1) gene, crucial for cell growth and differentiation, exhibits dysregulation linked to various cancer types, acting as both a positive and negative regulator. The selection of an appropriate method for the evaluation of gene promoter methylation status is important for clinical implementation without biases regarding false positive and false negative outcomes. The study focuses on the optimization of a novel droplet digital PCR (ddPCR) assay for identifying the methylation status of MEIS1. Compared to traditional methods, ddPCR offers an increased sensitivity and specificity, presenting a promising tool for precise DNA methylation assessment with potential implications for cancer diagnostics and prognostics.