Background/Objectives: Molecular profiling is essential for glioma classification, but in some cases tissue sampling may be limited, whereas reported detectability of cell-free tumor DNA (cftDNA) in plasma and cerebrospinal fluid (CSF) varies considerably. The aim of this study was to compare the detection rates and concentrations of cftDNA carrying IDH1 R132H and TERT promoter C228T/C250T mutations in paired plasma and CSF samples, as well as to examine associated clinical characteristics. Methods: 66 patients with WHO grade 1–4 gliomas were enrolled before surgery. Preoperative plasma and lumbar CSF were collected before tissue-damaging procedures. Cell-free DNA (cfDNA) was isolated from 5 mL of plasma and 2.5–5.0 mL of CSF and analyzed using mutation-specific droplet digital PCR (ddPCR) assays. Results: cfDNA was available from 65 CSF and 63 plasma samples. Total cfDNA concentration was lower in CSF than in plasma (medians: 434.1 vs. 1240.3 copies/mL; p < 0.001). Nevertheless, cftDNA carrying at least one of the assayed mutations was detected in 14/65 CSF samples (21.5%) and in 0/63 plasma samples (0.0%) (p < 0.001). IDH1 R132H was detected in 4.6% of CSF samples, and TERT promoter mutations were detected in 16.9%. Every CSF mutation with available paired tissue data was also identified in the corresponding tumor. Glioma grade 4 and contrast-enhancing tumors were associated with higher CSF cfDNA, cftDNA carrying at least one of the assayed mutations, and mutant allele fraction values (p < 0.05), whereas tumor size and localization were not significantly associated with these parameters (p > 0.05). Conclusions: With the presented targeted ddPCR workflow, lumbar CSF was substantially more informative than plasma for detecting the selected glioma-associated mutations. CSF cftDNA analysis may complement tissue profiling, although broader panels and larger prospective studies are needed to establish its clinical sensitivity.

Background: There is growing evidence that uterine microbiota might be linked to endometrial receptivity (ER) and affect the outcome of assisted reproductive technology (ART) procedures. Owing to the invasive nature of endometrial sampling, the evaluation of microbiota in this biomaterial is only possible outside the embryo transfer (ET) cycle. However, menstrual blood might be the key to overcoming this challenge as it can be safely aspirated from the uterine cavity at the beginning of the target ET cycle. This study aimed to evaluate the association of the microbiological profiles of menstrual blood with ER in patients undergoing frozen ET. Methods: Menstrual blood was obtained from 98 individuals scheduled for frozen ET in a private ART clinic (ET success rate–50%). DNA was isolated from menstrual sediment and analyzed using a multiplex quantitative PCR assay designed to identify 28 relevant microbial taxa and 3 Herpesviridae viruses. Results: Bacterial DNA was detected in 75.5% of samples. There were no associations between the abundance of individual microbial taxa and the outcome of ET, and the same was true for Shannon’s α-diversity indices (p > 0.05). However, Candida spp. and Enterobacteriaceae were detected exclusively in patients with negative ET outcomes (p = 0.028). Individuals with recurrent implantation failure had a significantly lower abundance of Lactobacillus spp. than the rest (0.0 [0.0; 7.4] vs. 2.8 [0.0; 91.9] %, p = 0.024). Conclusions: Menstrual blood aspirated directly from the uterus is a promising biomaterial for endometrial microbiological profiling. Upon further investigation, its analysis might become a useful tool in managing infertile patients scheduled for ART treatment.

Mosaic loss of the Y chromosome (mLOY) is a common karyotype alteration. The accumulation of LOY cells leads to genetic mosaicism and affects the development of pathological conditions in elderly men. Emerging data suggest that this phenomenon might be associated with an increased risk of cardiovascular disease (CVD) and several clinical and demographic parameters. However, the short-term dynamics of mLOY remain insufficiently investigated. Therefore, this study aimed to evaluate the short-term dynamics of mLOY in peripheral blood and its association with various features in men with high CVD risk. The study included healthy young volunteers (control group; n = 48) and male patients with a high risk of cardiovascular disease (HCR group; n = 65). Peripheral blood was obtained for the mLOY analysis from the HCR group at two time points: initially and after 6 months. mLOY was analyzed using a self-designed assay targeting SRY and EIF2C1 as a reference for digital droplet PCR. This assay demonstrated a high degree of linearity for Y chromosome copy numbers (CNs) from 0.9 to 0.6 (R² = 0.9986) in mLOY models. In the control group, the CNs varied from 0.9 to 1.1. mLOY was detected in 6 of 65 patients in the HCR group (CNs below 0.9). This study revealed preliminary, hypothesis-generating associations between mLOY and sedentary lifestyle and family history of early-onset CVDs (p < 0.05; however, the former did not retain significance after adjusting for age). Analysis of temporal variations of mLOY demonstrated heterogeneous dynamics: in some patients, mLOY resolved, whereas in others, Y chromosome CN decreased even further with time. However, given the low mLOY incidence rate in the investigated cohort, these results should be considered preliminary and exploratory. Further studies are warranted to determine the potential utility of mLOY and its temporal dynamics for assessing CVD risk.