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.

Recurrent wheezing represents a significant cause of respiratory morbidity in preschool children. While viral infections in the context of immune dysregulation are primary drivers, gastroesophageal reflux disease (GERD) and laryngopharyngeal reflux (LPR) are increasingly recognized as critical triggers. The link between GERD and respiratory symptoms has historically been controversial; however, recent advances necessitate a re-evaluation of this relationship. This review critically analyzes the pathophysiological nexus between reflux and airway hyperresponsiveness, emphasizing the dual mechanism of microaspiration and vagal reflex, while integrating the concept of reverse causality (where respiratory effort actively exacerbates reflux). We highlight that the “respiratory reflux” phenotype in preschoolers is often characterized by non-acid and proximal episodes, which standard pH-metry fails to detect. Consequently, we discuss the diagnostic shift towards multichannel intraluminal impedance-pH (MII-pH) monitoring and the incorporation of novel metrics defined by the Lyon Consensus 2.0 (MNBI, PSPW index), alongside specific biomarkers such as pepsin. Finally, we propose a phenotype-driven management algorithm, differentiating between acid-suppressive therapy and alginate-based interventions, to mitigate disease burden and improve clinical outcomes in refractory cases.

Bronchial asthma is the most common chronic respiratory disease in children. Treatment approaches should aim to achieve disease control, reduce limiting symptoms, and improve quality of life. The routine management of patients with asthma relies on the assessment of symptoms (including persistent cough and wheezing) and spirometry results However, these strategies do not consider the level of airway inflammation, which is a fundamental pathognomonic feature of the disease. The use of biomarkers is increasing in an effort to better understand individual asthma pathways (endotyping), offering the potential for personalized treatment with innovative biologics. Elevated levels of fractional exhaled nitric oxide (FeNO) are an indirect marker of T2 airway inflammation. FeNO is one of the few biomarkers that has been implemented in routine clinical practice because of its undeniable advantages, such as accessibility, noninvasiveness, simple implementation, and cost-effectiveness. However, the current scientific evidence in children is insufficient to recommend its universal use in the diagnosis and treatment of all patients. Considering its known limitations, FeNO can enhance therapeutic processes, particularly in children with severe asthma. High FeNO levels predict a good therapeutic response to corticosteroids and selected biologics (e.g., omalizumab and dupilumab) and indicate an increased risk of exacerbation. Similarly, measuring FeNO levels in children with recurrent wheezing can help determine the risk of future asthma development. Various factors contributing to FeNO value variability must be considered when interpreting the results. The lack of robust clinical studies is reflected in inconsistent guidelines from professional societies regarding the use of FeNO in managing childhood asthma. This review aims to evaluate the advantages, disadvantages, and potential applications of FeNO measurements in light of new findings in the field.

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.

METHODS:A total of 202 children aged 7-18 years were included in the study and stratified according to the treatment type and fractional exhaled nitric oxide (FeNO) values. Absolute peripheral blood eosinophil counts, nasal eosinophil percentages, FeNO levels and age were assessed. RESULTS:Antihistamines and leukotriene receptor antagonists were associated with a significant reduction in nasal eosinophilia (p = 0.004 and p = 0.001, respectively), whereas inhaled corticosteroids showed no significant effect (p = 0.928). A significant positive association was demonstrated between FeNO levels and nasal eosinophilia (p < 0.001), while age showed a significant negative effect (p < 0.001). CONCLUSION:Nasal cytology may represent an easily applicable, and ethically acceptable tool for noninvasive monitoring local eosinophilic inflammation in pediatric asthma. Nasal eosinophilia appears to reflect dynamic airway inflammatory changes more directly than peripheral blood eosinophils and may be useful adjunctive marker for monitoring treatment-related changes and guiding personalized therapy.

Background:Pneumocystis pneumonia (PCP) typically affects immunocompromised children, including those with HIV infection, those receiving cancer treatment, and those who have undergone transplantation. However, it also affects children from socioeconomically disadvantaged backgrounds. Objectives: This retrospective single center study aimed to describe the clinical characteristics of a cohort of paediatric patients with suspected PCP and to evaluate the clinical feasibility and diagnostic contribution of different sample types. Methods: Eligibility criteria included all paediatric patients presenting with clinical signs of pneumonia who underwent diagnostic testing for P. jirovecii based on clinical suspicion. Participant selection was based on a review of electronic and written medical records. This study specifically focused on the use of gastric aspirates, which were the most frequently collected samples in this population. A total of 277 patients between November 2017 and September 2024 underwent diagnostic tests, including real-time PCR analysis of laryngeal swabs, sputum, bronchoalveolar lavage, gastric aspirates, and pleural effusion. The study utilized descriptive statistics (frequencies, percentages, median, range) to summarize data; formal analytical statistical tests were not performed given the study's descriptive nature. Data on clinical outcomes were collected from medical records documenting patient follow-up post-treatment. Results: The diagnosis of PCP was confirmed in 36 patients (13%), ranging from 2 months to 2.5 years old, with 31 cases (86%) occurring in infants under 12 months. In our cohort, the diagnosis was most frequently established from gastric aspirates, which accounted for 29 of the 36 positive cases (80%), followed by bronchoalveolar lavage (8%), laryngeal swabs (6%), and sputum (6%). The predominant clinical presentation was prolonged respiratory deterioration with chronic cough and persistent refractory bronchoconstriction in 29 patients (81%). Fourteen patients (39%) experienced acute respiratory distress syndrome (RDS) and required additional oxygen and systemic corticosteroid therapy. Following appropriate treatment, clinical improvement, resolution of bronchial obstruction, and nutritional recovery were achieved. In the cohort of RT-PCR-positive cases, characteristics such as low socioeconomic status, malnutrition, poor sanitation, and exposure to tobacco smoke and combustion fumes were frequently observed, all of which are potential confounders or effect modifiers that may contribute to secondary immunodeficiency. Conclusions: The study's retrospective, single-centre design is subject to inherent limitations, such as potential selection bias. Although bronchoalveolar lavage is the ideal diagnostic method, its acquisition can be challenging in critically ill young infants. Our findings highlight the clinical utility of gastric aspirates as a valuable, minimally invasive alternative for diagnosing PCP in this vulnerable population.

Background/Objectives: Seasonal influenza imposes a significant burden on pediatric public health. Despite official recommendations and full insurance coverage, vaccination rates among children in Slovakia remain critically low. This study aims to analyze the attitudes, beliefs, and determinants of parental hesitancy regarding childhood influenza vaccination in the post-pandemic context. Methods: A single-center cross-sectional survey was conducted between February and March 2025 using convenience sampling among parents of children attending a pediatric immunoallergology center. An anonymous questionnaire collected data on demographics, risk perception, and attitudes. Data from 301 parents were analyzed using descriptive statistics, chi-squared tests, and odds ratios (OR) to identify key predictors of hesitancy. Results: Only 27.6% of parents expressed willingness to vaccinate their children, while 42.5% were opposed and 29.9% hesitant. Statistical analysis revealed no significant association between parental university education and vaccination intent (p > 0.05), indicating that vaccine hesitancy in this specific setting was present across all educational backgrounds. However, the source of information proved to be a critical determinant: consulting a pediatrician significantly increased the odds of acceptance (OR = 6.32; 95% CI: 3.54-11.28), whereas reliance on the internet and social media was a significant predictor of refusal (OR = 0.29; 95% CI: 0.17-0.50). The primary reported barrier was fear of adverse effects (70.4%), which significantly outweighed doubts about efficacy (30.2%). Conclusions: Parental hesitancy in Slovakia is a widespread phenomenon pervasive across all educational backgrounds, driven primarily by safety concerns and digital misinformation. The contrast between the protective influence of pediatricians and the negative impact of digital media underscores that clinical encounters are currently the most effective firewall against hesitancy. Public health strategies must therefore pivot from general education to empowering pediatricians with active, presumptive communication strategies.

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.

This case report presents a 7-year-old boy with severe, early-onset cow’s milk allergy (CMA) and poorly controlled, severe persistent asthma, highlighting the challenges of managing pediatric allergic multimorbidity. The patient experienced multiple anaphylactic reactions to cow’s milk, significantly impacting his quality of life. Despite adherence to conventional asthma therapies, his asthma remained poorly controlled. Given the severity of his condition and the underlying immunoglobulin E (IgE)-mediated inflammation, treatment with omalizumab, an anti-IgE monoclonal antibody, was initiated. Following omalizumab initiation, significant improvements were observed in asthma control, including reduced symptoms, improved pulmonary function, and decreased airway inflammation. Additionally, although a formal oral food challenge after five months of treatment elicited an anaphylactic reaction, the threshold dose triggering the reaction was significantly higher than previous reactions, indicating an increase in tolerance to cow’s milk proteins. This case underscores the potential of omalizumab in improving asthma control and increasing tolerance to cow’s milk, with associated benefits to the patient, but also highlights the variability in response when targeting co-existing allergic conditions. Further research is needed to optimize treatment strategies for pediatric allergic multimorbidity.

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.

Specific allergen immunotherapy (AIT) represents the only causal treatment for allergic diseases capable of modifying the natural disease trajectory and inducing long-term allergen tolerance. Its sublingual form (SLIT) has emerged as a safe and highly effective alternative to conventional subcutaneous administration. In recent years, scientific interest has increasingly focused on the complex interplay between SLIT and the human microbiome. We conducted a targeted literature search across four major electronic databases (PubMed/MEDLINE, Scopus, Web of Science, and the Cochrane Library) from January 2012 to February 2026. Applying strict inclusion criteria, a narrative review approach underpinned by a systematic search strategy was utilized to qualitatively synthesize 41 peer-reviewed articles evaluating baseline dysbiosis, post-SLIT microbial shifts, and adjuvant interventions. A growing body of evidence suggests that the therapeutic efficacy of SLIT is associated with alterations in microbial communities across the oral–gut–immune axis. However, recent clinical data present a more nuanced picture, indicating that clinical efficacy can occur without complete microbial normalization. Integrating these insights with established immunology highlights the microbiome’s role as an epigenetic stabilizer in maintaining rather than merely initiating long-term allergen tolerance. Shifting toward precision allergology, we critically appraise the search for predictive baseline biomarkers, expose the severe methodological heterogeneity limiting current probiotic adjuvant trials, and underscore the unexplored potential of dietary interventions (e.g., fermentable fibers) to harness endogenous tolerogenic metabolites.

Background Chronic cough is a frequent symptom of pediatric Long COVID, hypothetically driven by viral neurotropism and sensory nerve sensitization. We investigated the neuro-immune axis in pediatric asthma to determine if the magnitude of post-SARS-CoV-2 humoral immunity correlates with objective airway afferent nerve hypersensitivity. Methods This prospective observational study included 61 pre-pubertal children (aged 8 to < 12 years) with well-controlled, predominantly inhaled corticosteroid (ICS)-treated (93.4%) bronchial asthma and confirmed past SARS-CoV-2 infection. Systemic humoral memory was quantified via anti-Spike IgG and IgA titers. Objective cough reflex sensitivity was measured using a capsaicin challenge test, establishing C2 and C5 values. Subjective symptom burden was evaluated using parent-proxy questionnaires (PCQ, VAS, PedsQL). Results Stratification by median anti-Spike IgG (125.77 BAU/ml) revealed no significant differences in basal (C2, p = 0.301) or motor response (C5, p = 0.714) capsaicin thresholds between robust and waning humoral memory states. IgA stratification yielded identical results. Spearman's correlation confirmed a complete lack of association between absolute IgG titers and neurophysiological markers (p > 0.05). Crucially, parent-reported chronic cough severity (PCQ, VAS) and asthma-specific quality of life demonstrated a complete dissociation from objective capsaicin thresholds across all evaluated domains (all p > 0.05). Supplementary subgroup analysis revealed no significant differences in cough thresholds based on acute COVID-19 severity (p > 0.05). Conclusion A robust post-viral humoral immune response to SARS-CoV-2 does not precipitate peripheral airway nerve hypersensitivity in properly controlled, ICS-treated asthmatic children. The complete uncoupling of subjective parent-reported symptoms from objective neurophysiology cautions against diagnosing neurogenic Long COVID based solely on questionnaires, emphasizing the necessity of objective testing and evaluation of alternative atopic etiologies.

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.

Up-to-date data on roles of ATP‑sensitive potassium (KATP) channels indicate their emerging roles in neurodegeneration. The aim of present study was to evaluate the significance of KATP channels on cell viability, calcium dynamics, and mitochondrial morphology with the accent on their intracellular localization. We distinguished between whole-cell KATP effects and specific effects of mitochondrial KATP under both physiological conditions and pathological conditions simulating in vitro Parkinson´s-type neurodegeneration. SH‑SY5Y cells with its high fidelity to dopaminergic neurons were treated for 24 h with the non‑selective KATP opener pinacidil and blocker glibenclamide, or with the mitochondrial KATP opener diazoxide and blocker 5‑hydroxydecanoate (5HD). The effects of modulators were analysed alone or alongside with rotenone, which is widely used as an inducer of Parkinson´s-type neurodegeneration. Intracellular calcium distribution and mitochondrial rebuild pattern was evaluated using the cell segmentation performed by fluorescent confocal microscopy. Although none of the KATP modulators reversed the negative effects of rotenone, significant and selective effects of mitochondrial KATP modulation on calcium homeostasis and mitochondrial morphology were observed. For antagonists, both compounds showed consistent effects, with non-selective glibenclamide exerting stronger effects, particularly in elevating calcium. More distinctive results were obtained for agonists: both reduced calcium concentration; however, pinacidil tended to induce mitochondrial fragmentation, an effect absent in diazoxide-treated cells. Furthermore, strong correlations were identified between calcium levels and several mitochondrial and cell viability parameters.

Background: Acute respiratory infections (ARIs) pose a significant clinical challenge in paediatric populations, especially in children with comorbidities who may exhibit underlying immune dysregulation. Inosine pranobex (IP) is an immunomodulatory agent that enhances T-lymphocyte and Natural Killer (NK) cell function, offering a targeted therapeutic rationale for such cases. Objective: This study aimed to retrospectively describe the clinical characteristics, immunological profiles, and outcomes of paediatric patients with complex, PCR-confirmed viral ARIs and significant comorbidities, for whom adjunctive therapy with IP was initiated based on clinical judgment. Methods: This retrospective case series analysed data from 14 paediatric patients hospitalised at a specialised centre (National Institute of Paediatric Tuberculosis and Respiratory Diseases in Dolny Smokovec, Slovakia). Cases were selected based on PCR-confirmed viral ARI, a history of recurrent infections, significant comorbidities, and initiation of IP therapy. The indication for IP was guided by the treating physician in cases of severe, prolonged, or recurrent disease course, where immune dysregulation was suspected, often supported by prior immunophenotyping. Results: A frequent observation in this cohort was the presence of baseline cellular immune alterations with a frequent observation of baseline cellular immune alterations, most notably the depletion of natural killer (NK) cells. NK cell depletion was identified in half of the patients (7/14). Following the initiation of treatment regimens that included adjunctive IP, clinical stabilisation or improvement was observed in all 14 patients included in the study. The therapy was well tolerated, with no reported adverse events attributable to IP. Conclusions: This case series highlights the common presence of cellular immune alterations in children with complex ARIs. While the observational nature of this study precludes any conclusions about causality, the favourable clinical course, safety profile, and strong immunological rationale support the need for prospective controlled trials to evaluate the role of IP in this specific high-risk paediatric population.