Maciej Skrzypek

Cancer risk may be influenced by genetic variation and altered expression of xenobiotic-metabolizing enzymes, yet their role in endometrial cancer remains incompletely understood. This study evaluated the association between four polymorphisms in xenobiotic metabolism-related genes CYP1A1 rs1799814, CYP2D6 rs3892097, NAT1 rs72554606, and NAT2 rs1799930 and the risk of endometrial cancer, and assessed CYP1A1 and CYP2D6 expression in tumor and control tissues. Genetic association analyses, including multivariate and histology-stratified models, were performed, and gene expression levels were compared between cancer and control tissues. Variants in NAT2, CYP1A1, and CYP2D6 were significantly associated with an increased risk of endometrial cancer, whereas NAT1 rs72554606 showed a protective effect, particularly in the dominant model. The strongest association was observed for NAT2 rs1799930 in additive and recessive models. Expression analysis revealed significantly higher CYP1A1 and CYP2D6 levels in tumor tissues than in control tissues. Stratified analyses showed generally consistent effects, especially for endometrioid carcinoma, although estimates for the serous subtype were limited by sample size. These findings suggest that polymorphisms and altered expression of xenobiotic-metabolizing genes may contribute to endometrial carcinogenesis. Further studies, including independent validation and analyses of gene–environment interactions, are needed.

Bladder cancer (BC) is among the top ten most commonly diagnosed cancers worldwide, with rising incidence and around 213,000 deaths annually. It exhibits a marked male predominance, largely attributable to risk factors such as tobacco use, occupational carcinogen exposure, and genetic predisposition. Bladder cancer is classified into non-muscle-invasive (NMIBC) and muscle-invasive (MIBC) forms, with the latter linked to worse prognosis. Despite current treatment modalities, including surgical resection with adjuvant chemotherapy or immunotherapy, disease management remains challenging due to high recurrence rates and limited therapeutic efficacy in advanced stages. Recent studies highlight telomerase activity, especially TERT promoter mutations present in about 70% of BC cases, as a key contributor to disease progression. These findings position telomerase as a promising biomarker for early detection, disease monitoring, and targeted therapy. This review summarizes recent advances in understanding telomerase regulation in BC and its potential clinical applications.

Exosomes are small extracellular vesicles found in body fluids. They contain proteins and nucleic acids that reflect the condition of the parent cell. Because of this, they may be useful in colorectal cancer (CRC) diagnostics. In this study we compared three plasma isolation methods: ultracentrifugation (UC), size exclusion chromatography (SEC), and a commercial Exo-spin kit. The material obtained with each method was checked for protein content, purity, and vesicle integrity using protein measurements, Western blots, and scanning electron microscopy. From these results SEC was chosen for further use. Exosomes isolated with this method were tested by an enzyme-linked immunosorbent assay (ELISA) for four CRC-related proteins: HSP70, CK19, CA125, and TAG72. Patient samples showed higher levels of HSP70, CK19, and CA125 than controls. TAG72 levels did not differ. In addition, men had more HSP70 than women. These findings show that SEC can be applied for exosome isolation from plasma and that specific proteins detected in exosomes, including HSP70, CK19, and CA125, may serve as promising markers for non-invasive colorectal cancer diagnosis and monitoring.

Rheumatoid arthritis (RA) is a chronic, common autoimmune disease. It is characterized by inflammatory polyarthritis, which can lead to permanent disability in patients. Current treatment is mainly symptom-related, aiming to reduce pain and inflammation, but does not lead to a full recovery. This treatment includes non-steroidal anti-inflammatory drugs (NSAIDs) and disease-modifying anti-rheumatic drugs (DMARDs). It has been shown that, due to chronic inflammation, reduced glucose levels and hypoxia, endoplasmic reticulum (ER) stress is induced in RA patients, leading to the activation of multiple signaling pathways, including the ER-dependent adaptation of the unfolded protein response (UPR) pathway. The aim of this study was to assess the level of apoptosis in patients diagnosed with RA. The study sought to investigate whether UPR response correlated with apoptosis induction could serve as a potential diagnostic marker or therapeutic target. In vitro studies have shown that UPR pathway activity can be observed in patients diagnosed with RA. The study group consisted of PBMC cells from 61 individuals, including a total of 31 rheumatoid arthritis patients and 30 healthy controls. In order to validate UPR activation, we estimated molecular markers of ER stress via RT-qPCR expression analysis. GAPDH expression was used as a standard control. Elevated levels of mRNA for the eIF2α (p-value = 0.001903), the BBC3 (PUMA) (p-value = 0.007457 × 10⁻⁷) and the TP53 (p-value = 0.002212) were confirmed in a group of RA patients. Further analysis showed that after the induction of apoptosis the percentage of DNA contained in the tail was 37.78% higher in RA patients than in the control group (p-value = 0.0003) measured by comet assay. The exogenous damage caused by hydrogen peroxide was found to be statistically elevated in RA patients and the caspase-3 level was calculated of 40.17% higher than in controls (p-value = 0.0028). It was also found that PBMC cells from RA patients were more sensitive to apoptotic induction. Our results were confirmed by flow cytometry. The most important finding from our data was the confirmation of elevated sensitivity to apoptosis induction in RA patients; the results showed a 40.23% higher percentage of cells in early apoptosis than in the control group (p-value = 0.0105). Our results may help to assess the feasibility of the application of early diagnosis and targeted therapy in the treatment of RA patients, including the ER signaling pathway via selected UPR-dependent molecular inhibitors.

Peripheral nerve regeneration remains a major challenge in neuroscience, despite advancements in understanding its mechanisms. Current treatments, including nerve transplantation and drug therapies, face limitations such as invasiveness and incomplete recovery of nerve function. Physical therapies, like pulsed electromagnetic fields (PEMF) and low-intensity ultrasound (LIPUS), are gaining attention for their potential to enhance regeneration. This study analyzes the effects of PEMF and LIPUS on gene expression in human primary Schwann cells, which are crucial for nerve myelination and repair. Key genes involved in neurotrophin signaling (NGF, BDNF), inflammation (IL-1β, IL-6, IL-10, TNF-α, TGF-β), and regeneration (CRYAB, CSPG, Ki67) were assessed. The results of this study reveal that combined PEMF and LIPUS therapies promote Schwann cell proliferation, reduce inflammation, and improve the regenerative environment, offering potential for optimizing these therapies for clinical use in regenerative medicine.

Although the epidemiology and symptoms of major depressive disorder (MDD) have been well-documented, the etiology and pathophysiology of the disease have not yet been fully explained. Depression arises from intricate interplay among social, psychological, and biological factors. Recently, there has been growing focus on the involvement of miRNAs in depression, with suggestions that abnormal miRNA processing locally at the synapse contributes to MDD. Changes in miRNAs may result from altered expression and/or function of the miRNA biogenesis machinery at the synapse. The aim of our research was to assess the relationship between the occurrence of depression and single-nucleotide polymorphisms (SNP) in the following genes in the Polish population: DROSHA (rs6877842; rs10719) and XPO5 (rs11077). This study involved 200 individuals, including 100 with depressive disorders in the study group (SG) and 100 healthy people without MDD in the control group (CG). All participants were unrelated native Caucasian Poles from central Poland. Blood samples were collected to evaluate the single-nucleotide polymorphism of the genes. Findings indicated that within our patient cohort, the risk of depression is increased by polymorphic variants of the rs10719/DROSHA and rs11077/XPO5 genes and lowered by rs6877842/DROSHA. Our study sheds light on the understanding of the genetic basis of depression, which can be used in the rapid diagnosis of this disease.

Rheumatoid arthritis (RA) is a chronic inflammatory disease. Despite new methods of diagnostics and treatment as well as extensive biological and immunosuppressive treatment, the etiology of RA is not fully understood. Moreover, the problem of diagnosis and treatment of RA patients is still current and affects a large group of patients. It is suggested that endoplasmic reticulum (ER)-related features may impair adaptation to chronic stress, inferring the risk of rheumatoid arthritis. The main goal in this study was evaluation of changes in mRNA translation to determine chronic ER stress conditions in rheumatoid arthritis patients. The study group consist of 86 individuals including a total of 56 rheumatoid arthritis patients and 30 healthy controls. The expression level of mRNA form blood samples of RA patients as well as controls of the unfolded protein response (UPR)-associated genes (p-eIF2, BCL-2, PERK, ATF4, and BAX) were investigated using real-time qPCR. GAPDH expression was used as a standard control. Considering the median, the expression levels of PERK, BCL-2, p-eIF2, ATF4, and BAX were found to be significantly increased in the blood of RA patients compared with the control group. The p-value for the PERK gene was 0.0000000036, the p-value for the BCL-2 gene was 0.000000014, the p-value for the p-eIF2 gene was 0.006948, the p-value for the ATF4 gene was 0.0000056, and the p-value for the BAX gene was 0.00019, respectively. Thus, it can be concluded that the targeting of the components of the PERK-dependent UPR signaling pathway via small-molecule PERK inhibitors may contribute to the development of novel, innovative treatment strategies against rheumatoid arthritis.

Rheumatoid arthritis (RA) is a chronic inflammatory disease. Despite new methods of diagnostics and treatment as well as extensive biological and immunosuppressive treatment, the etiology of RA is not fully understood. Moreover, the problem of diagnosis and treatment of RA patients is still current and affects a large group of patients. It is suggested that these ER-related features may impair adaptation to chronic stress, inferring the risk of rheumatoid arthritis. The main goal in this study was evaluation of changes in mRNA translation to determine chronic ER stress conditions in rheumatoid arthritis patients. The study group consist of 86 individuals including total 56 rheumatoid arthritis patients and 30 healthy controls. The expression level of mRNA form blood samples of RA patients as well as controls of the PERK-UPR associated genes: p-eIF2, BCL-2, PERK, ATF4, BAX were investigated by Real-Time qPCR. GAPDH expression was used as a standard control. Considering the median, the expression of PERK, BCL-2, p-eIF2, ATF4, BAX was found to be significantly increased in the blood of RA patients compared with the control group. The p-value for the PERK gene was 0.0000000036, the p-value for the BCL-2 gene was 0.000000014, the p-value for the p-eIF2 gene was 0.006948, the p-value for the ATF4 gene was 0.0000056 and the p-value for the BAX gene was 0.00019, respectively. Thus, it can be concluded that targeting of the components of the PERK-dependent UPR signaling pathway via small-molecule PERK inhibitors, may contribute to the development of novel, innovative treatment strategies against rheumatoid arthritis.

Background: Single nucleotide polymorphisms in gene encoding is the key enzyme in the folates pathway, methyltetrahydrofolate reductase (MTHFR), which causes methylation disorders associated with coronary artery disease (CAD). We evaluated associations between methylation disorders caused by MTHFR gene polymorphisms and the blood folate concentrations (folic acid, 5-MTHF) in CAD patients. Methods: Study group: 34 patients with CAD confirmed by invasive coronary angiography (ICA). Controls: 14 patients without CAD symptoms or significant coronary artery stenosis, based on ICA or multislice computed tomography (MSCT) with coronary artery calcification (CAC) scoring. Real-time PCR genotyping was assessed using TaqMan™ probes. Folic acid and 5-MTHF concentrations in blood serum were determined using Liquid Chromatography-Mass Spectrometry (LC-MS). Results: The c.[1286A>C];[1286A>C] MTHFR polymorphism occurred significantly more often in (CAD⁺) patients compared to the (CAD⁻) cohort and to the selected general European “CEU_GENO_PANEL” population sample. The concentration of 5-MTHF and folic acid in subgroups of CAD⁺ patients with methylation disorders categorized by genotypes and CAD presence (CAD⁺) was always lower in CAD⁺ subgroups compared to non-CAD individuals (CAD⁻). Conclusions: Further studies on a larger scale are needed to implicate the homozygous c.1286A>C MTHFR variant as CAD genetic marker and the 5-MTHF as CAD biomarker. Identification of high CAD risk using genetic and phenotypic tests can contribute to personalized therapy using an active (methylated) form of folic acid (5-MTHF) in CAD patients with MTHFR polymorphisms.

Background: The presence of single nucleotide polymorphisms in the gene encoding key en-zyme in the folate pathway methyltetrahydrofolate reductase (MTHFR), causes methylation disorders associated with CAD development. Methods: Study group: 42 patients with CAD con-firmed by coronary angiography. Controls: 16 patients without CAD symptoms or significant coronary artery stenosis, based on invasive coronary angiography or multislice computed to-mography with coronary artery calcification scoring. Real-time PCR genotyping was assessed using TaqMan™ probes. Folic acid and 5-MTHF concentrations in blood serum were determined by LC-MS. Results: c.[1286A>C];[1286A>C] MTHFR polymorphism occurred significantly more often in (CAD+) patients compared to (CAD-) cohort and to the European population. The con-centration of 5-MTHF and folic acid in the subgroups of patients with methylation disorders di-vided by genotypes and (CAD+) was lower compared to (CAD-). Conclusions: The homozygous c.1286 A>C MTHFR variant can be considered as CAD genetic marker. In the subjects with the c.1286 A>C heterozygous and homozygous genotypes, lower concentrations of methyltetrahy-drofolate (5-MTHF) were observed. 5-MTHF can be considered as a biomarker of CAD. Identifi-cation of people at high risk of CAD using genetic tests can contribute to personalized therapy using active (methylated) form of folic acid (5-MTHF) in CAD patients with MTHFR polymor-phisms.