Aim: This study aimed to determine the associations of MEG3 rs7158663, rs4081134 gene variants, as well as the immunohistochemical markers Ki-67, p53, and CK18, with the clinical features of pituitary neuroendocrine tumors (PitNETs).Methods: This case-control study included 340 individuals who were divided into two groups: a control group (n=220) and a PitNETgroup (n=120). DNA was isolated from the venous blood of study participants by the leukocyte salt precipitation method. Real-time polymerase chain reaction was used for the MEG3 rs7158663, rs4081134 single nucleotide variants genotyping. Immunohistochemical analysis of Ki-67 labeling index and p53 protein biomarkers was performed using the automated Ventana BenchMark ULTRA PLUS staining system, following the manufacturer’s recommendations. CK18 immunostaining was conducted with the Dako Omnis staining system, following the manufacturer’s recommendations. Monoclonal antibodies SP6, DO-7, and DC-10 were used to detect Ki-67 labeling index, p53, and CK18, respectively. Statistical data analysis was performed using the “IBM SPSS Statistics 30.0” program.Results: Genotype and allele frequencies of MEG3 rs7158663 and rs4081134 variants showed no significant differences between healthy controls and PitNET patient groups. Additionally, no associations were found between either MEG3 variants and PitNET recurrence, size, invasiveness, and functional status. Ki-67 labeling index (>3% vs. ≤3%) showed no significant differences with any clinical feature of PitNETs (recurrence, size, invasiveness, functional status). In contrast, the p53 H-score was significantly higher in macroadenomas than in microadenomas (median 27 vs. 16; p=0.008). Additionally, invasive pituitary adenomas showed a higher p53 H-score compared with non-invasive tumors (median 27 vs. 20; p=0.018). Negative CK18 immunostaining was significantly more frequent in invasive than non-invasive PitNETs (44.4% vs. 13.3%; p < 0.001) and in non-functioning compared to functioning adenomas (42.0% vs. 18.4%; p=0.011). No significant associations were found between either MEG3 variant and Ki-67 LI, p53 H-score, or CK18 immunohistochemical reactions.Conclusions: This study found that a higher p53 H-score was significantly associated with larger PitNET size and invasiveness. Negative CK18 staining was associated with non-functioning and invasive PitNETs. P53 expression and CK18 status may serve as useful prognostic markers in PitNETs.

Background and Objectives: Age-related macular degeneration (AMD) is the leading cause of blindness, affecting millions worldwide. Its pathogenesis involves the death of the retinal pigment epithelium (RPE), followed by photoreceptor degeneration. Although AMD is multifactorial, various genetic markers are strongly associated with the disease and may serve as biomarkers for evaluating treatment efficacy. This study investigates TAB2 rs237025, IKBKB rs13278372, and IKBKG rs2472395 variants and their respective serum protein concentrations in relation to AMD occurrence and exudative AMD treatment response to anti-VEGF treatment. Materials and Methods: The case–control study involved 961 individuals, and they were divided into three groups: control, early AMD, and exudative AM patients. Genotyping of selected SNPs were conducted using a real-time polymerase chain reaction method (RT-PCR). Based on the clinical OCT and BCVA data, patients with exudative AMD were categorized into one of two groups: responders and non-responders. The data obtained were analyzed using the “IBM SPSS Statistics 29.0” software program. Results: Our study revealed that TAB2 rs237025 allele A was identified as a risk factor for early and exudative AMD development. The same associations remained only in females with exudative AMD but not in males, suggesting gender-specific pathogenetic pathways in exudative AMD. Analysis of IKBKB rs13278372 or serum IKBKB protein associations with early or exudative AMD occurrence in the Lithuanian population revealed no significant associations. On the other hand, we found that each A allele of IKBKB rs13278372 was associated with a worse response to anti-VEGF treatment (OR = 0.347; 95% CI: 0.145–0.961; p = 0.041). These results suggest a potential marker for future studies evaluating anti-VEGF treatment for exudative AMD patients. IKBKG rs2472395 was a protective variant for early AMD in males and for exudative AMD in females only. Also, IKBKG protein concentration was lower in exudative AMD relative to the control group (median (IQR): 0.442 (0.152) vs. 0.538 (0.337), p = 0.015). Moreover, exudative AMD patients who carry the GG genotype of IKBKG rs2472394 exhibited significantly reduced serum IKBKG concentrations compared to the controls (median (IQR): 0.434 (0.199) vs. 0.603 (0.335), p = 0.012), leading to the hypothesis that the IKBKG rs2472394 variant might play a role in protein concentration differences and exudative AMD development. Conclusions: Our study identified the TAB2 rs237025 allele A as a significant risk factor for both early and exudative AMD, with gender-specific associations observed in females with exudative AMD, suggesting distinct pathogenetic pathways. While IKBKB rs13278372 and serum IKBKB protein levels showed no significant association with AMD development, the A allele of IKBKB rs13278372 was associated with a worse response to anti-VEGF treatment, indicating its potential as a marker for treatment outcomes. Additionally, the IKBKG rs2472395 variant was found to be protective for early AMD in males and exudative AMD in females, and lower IKBKG protein levels were associated with exudative AMD, particularly in patients with the GG genotype of IKBKG rs2472394, suggesting its role in protein concentration and disease progression. These findings highlight genetic markers that may contribute to AMD pathogenesis and treatment response.

The study aimed to investigate the association between the TAS2R16 gene (rs860170, rs978739, rs1357949), TAS2R16 serum levels, and multiple sclerosis (MS). A total of 265 healthy control subjects and 218 MS patients were included in the study. Single nucleotide polymorphisms (SNPs) were tested by real-time polymerase chain reaction (RT-PCR). The serum concentration of TAS2R16 was measured using the ELISA method. Analyses revealed that the TAS2R16 rs860170 TT genotype was statistically significantly less frequent in the MS group than in the control group (p = 0.041), and the CC genotype was statistically significantly more frequent in the MS group than in the control group (p < 0.001). In the most robust (codominant) model, the CC genotype was found to increase the odds of MS by ~27-fold (p = 0.002), and each C allele increased the odds of MS by 1.8-fold (p < 0.001). Haplotype analysis of the rs860170, rs978739, and rs1357949 polymorphisms showed that the C-C-A haplotype was associated with a ~12-fold increased odds of MS occurrence (p = 0.02). Serum TAS2R16 levels were elevated in the MS group compared to control subjects (p = 0.014). Conclusions: The rs860170, rs978739, and rs1357949 polymorphisms demonstrated that the C-C-A haplotype and elevated TAS2R16 serum levels can promote the development of MS. These preliminary findings underscore the importance of specific genetic variants, such as rs860170, rs978739, and rs1357949, in MS risk. Additionally, elevated TAS2R16 serum levels in MS patients suggest a potential role in MS pathogenesis. These findings provide insights into the genetic and molecular mechanisms underlying MS and pave the way for personalized diagnostic and therapeutic strategies. Integrating genetic and serum biomarker data in MS research offers promising avenues for improving clinical outcomes and advancing precision medicine approaches in the future.

According to recent research, inflammatory STAT4 and its protein impact may be important factors in developing cancerous diseases. Still unanalyzed is this effect in patients with laryngeal squamous cell carcinoma (LSCC). In the present study, we evaluated four single nucleotide variants (SNVs) of STAT4 (rs10181656, rs7574865, rs7601754, and rs10168266) and STAT4 serum levels to determine their link between LSCC development and its clinical manifestations. A total of 632 men (324 LSCC patients and 338 healthy individuals) were involved in this study. The genotyping was carried out using real-time PCR. Additionally, we measured 80 study subjects’ (40 LSCC patients and 40 control subjects) STAT4 protein concentrations using an enzyme-linked immunosorbent assay (ELISA). In our study, the T allele of STAT4 rs7574865 significantly increases the likelihood of LSCC occurrence by 1.4-fold. Additionally, this SNV is associated with higher odds of early-stage disease, T1 size LSCC development, absence of metastasis to neck lymph nodes, and well-differentiated carcinoma. The G allele of rs10181656 is significantly associated with various clinical characteristics of LSCC, increasing the odds of early- and advanced-stage disease by 2.8-fold and 1.9-fold, respectively. Additionally, this allele is linked to an increased likelihood of developing tumors of different sizes and non-metastasized LSCC, as well as poorly differentiated carcinoma, highlighting its potential impact on the development and features of LSCC. Conclusion: The analysis of the STAT4 rs7574865 SNV revealed that the G allele is linked to a more favorable prognosis in LSCC. Additionally, it is hypothesized that the G allele of rs10181656 may be associated with the occurrence of LSCC but may not serve as a sensitive prognostic biomarker for distinguishing between disease stages, cell differentiation, or tumor size.

Laryngeal squamous cell carcinoma (LSCC) is a common and aggressive head and neck malignancy with poor prognosis due to late diagnosis and limited biomarkers. Angiogenesis, driven by vascular endothelial growth factor A (VEGFA), plays a key role in tumor growth and metastasis. Single nucleotide variants (SNVs) in the VEGFA gene have been linked to various cancers, but their role in LSCC remains unclear. In this study, we investigated the association between VEGFA (rs1570360, rs699947, rs3025033, rs2146323, rs3024997) SNV and LSCC susceptibility in a large cohort of 297 LSCC patients and 390 age- and sex-matched control subjects. The DNA samples were obtained from peripheral blood leukocytes, which were purified using the DNA salting-out technique. Real-time polymerase chain reaction assessed SNVs. The data obtained were processed using the IBM SPSS Statistics 29.0 software. Using a detailed subgroup analysis based on tumor stage, size, metastasis, and differentiation grade, we assessed the potential links between VEGFA (rs1570360, rs699947, rs3025033, rs2146323, rs3024997) variants and LSCC development. Contrary to previous studies suggesting significant associations between VEGFA genetic variances with other squamous head and neck cell cancers, our study found no statistically significant differences in the distribution of these SNVs between LSCC patients and controls. Our findings suggest that VEGFA SNV may not be a key factor in LSCC susceptibility. However, the complexity of LSCC’s genetic underpinnings warrants further investigation with larger cohorts and additional genetic markers. Our results contribute to the ongoing debate on the role of angiogenesis in LSCC and provide insights into the challenges of identifying reliable biomarkers for this malignancy.

Background: Pituitary adenoma (PA) is a common benign tumor that develops in the pituitary gland, causing hormonal imbalances and potential health issues. The TAS2R16 gene codes for a taste receptor and is involved in bitter taste perception, but there is currently no known direct link between this gene and pituitary adenoma. Methods: This study included 221 healthy controls and 131 patients with pituitary adenoma (PA) from the Lithuanian population. DNA was isolated from peripheral venous blood using the salt precipitation method. Genotyping was performed via RT-PCR. Statistical analysis was conducted with IBM SPSS Statistics 29.0 software, incorporating the Bonferroni correction for multiple comparisons. Results: This study found that the TAS2R16 rs978739 C allele is less common in the non-invasive PA group compared to the control group (p = 0.045). The TAS2R16 rs860170 CT genotype reduces the likelihood of developing non-invasive PA by 1.9-fold under the codominant (p = 0.024) and overdominant (p = 0.030) models. The odds of developing non-invasive PA are reduced by 2-fold under the dominant (p = 0.021) model for TAS2R16 rs860170 CT + CC genotypes and by 2-fold under the additive (p = 0.018) model for each TAS2R16 rs860170 C allele. The PA group had higher serum levels of TAS2R16 than the control group (p < 0.001). The present study found that patients with the TAS2R16 rs978739 TT or CT genotype had higher serum TAS2R16 levels and protein concentrations than healthy individuals (p = 0.025 and p = 0.019, respectively), and those with the AA or AG genotype of TAS2R16 rs1357949 had higher protein concentrations (p = 0.005 and p = 0.007, respectively). Conclusions: The TAS2R16 rs978739 C allele was less common in the non-invasive PA group compared to the control group, while the TAS2R16 rs860170 CT genotype was linked to a reduced likelihood of developing non-invasive PA. Additionally, the PA group showed higher serum levels of TAS2R16, and increased serum protein concentrations were observed in PA patients with specific TAS2R16 variants.