Background. Pancreatic ductal adenocarcinoma remains difficult to treat and has an exceptionally poor prognosis. While prior studies have noted antitumour properties for midazolam in pancreatic cancer, whether this response involves developmental signaling pathways is unclear. Here, we evaluated midazolam-treated MIA PaCa-2 cells to identify associated Wnt and Hedgehog transcriptional shifts and their systems-level context. Methods. MIA PaCa-2 cells were exposed to midazolam (20–100 µM) for 24 h. We measured metabolic viability by MTT and apoptosis by Annexin V-FITC/7-AAD flow cytometry at 60 µM. Expression of 18 targeted genes was quantified by RT-qPCR using ACTB for normalization. Changed genes were examined through STRING/KEGG networks, TCGA-PAAD survival analysis, DGIdb annotations, and preliminary FZD7 molecular docking. Results. Midazolam reduced MTT-derived metabolic viability in a concentration-dependent manner (IC₅₀ = 54.6 µM) and significantly increased Annexin V-positive apoptotic populations at 60 µM across three independent experiments. Eight transcripts changed significantly: CTNNB1, TGFBR2, WNT2, WNT7B, HHIP, and DHH declined, while CSNK1A1 and AXIN1 increased (all adjusted p < 0.05). Network analysis highlighted CTNNB1 as the main hub, and high CTNNB1 expression correlated with shorter survival in TCGA-PAAD patients (p = 0.043; HR = 1.58, 95% CI: 1.01–2.47). Conclusions. Midazolam reduced metabolic viability and increased apoptosis in MIA PaCa-2 cells alongside coordinated expression changes in key Wnt and Hedgehog components. Because the required doses far exceed clinical sedation levels and pathway activity was not directly confirmed at the protein level, these findings should be regarded as a preliminary hypothesis rather than evidence of therapeutic efficacy.

Background: Colorectal cancer (CRC) is one of the most prevalent malignancies worldwide and remains a major health concern due to its high recurrence and mortality rates. Recent studies suggest that anesthetic agents, including ketamine, may have direct effects on cancer cell viability and apoptosis. Objective: This study aimed to investigate the in vitro effects of ketamine on the HT-29 human colorectal adenocarcinoma cell line, focusing on its cytotoxic and pro-apoptotic potential. Material and Methods: HT-29 cells were treated with ketamine for 24 h. Cell viability was evaluated using the MTT assay. Apoptosis rates were determined by flow cytometry with Annexin V-FITC/7-AAD staining. Furthermore, quantitative PCR (qPCR) was performed to assess the expression levels of key genes associated with proliferation and apoptosis. GeneQuery™ Human Basal Cell Carcinoma qPCR Array Kit (GQH-BCC-GK015-C) was used for qPCR analysıs. Molecular docking simulations were performed to investigate the potential molecular interactions between ketamine and three target proteins: the N-methyl-D-aspartate (NMDA) receptor, epidermal growth factor receptor (EGFR), and casein kinase 1 delta (CSNK1D). To ensure robustness of predictions, two independent docking methods were employed. Results: Ketamine significantly reduced cell viability in a dose-dependent manner, with an IC₅₀ value of approximately 1.05 µM. Flow cytometry analysis demonstrated a marked increase in early apoptosis (23.9%) in treated cells. These findings suggest that ketamine exhibits potential anti-proliferative and pro-apoptotic effects on HT-29 colorectal cancer cells. Conclusions: These findings suggest that ketamine exhibits potential anti-proliferative and pro-apoptotic effects on HT-29 colorectal cancer cells in vitro. Further studies are warranted to elucidate the underlying molecular mechanisms and potential clinical implications.

Pancreatic ductal adenocarcinoma (PDAC) remains a highly lethal malignancy, and there is continuing interest in whether agents used during the perioperative period influence tumour-cell biology. Propofol has been associated with antiproliferative and pro-apoptotic effects in experimental cancer models; however, its relationship with Wnt–Frizzled-related transcriptional responses in pancreatic cancer cells remains incompletely characterised. MIA PaCa-2 cells were exposed to propofol for 24 h. Cellular metabolic activity was assessed using the MTT assay over a concentration range of 0–20 µM, and apoptosis was evaluated via Annexin V-FITC/7-AAD flow cytometry at the MTT-derived IC₅₀ concentration of 7.6 µM. Transcript abundance of 18 pathway-associated genes was analysed via RT-qPCR following exposure to 7.6 µM propofol. MTT, flow-cytometry, and RT-qPCR assays were each performed in three independent biological experiments (n = 3). RT-qPCR results were evaluated after Benjamini–Hochberg false discovery rate correction across the 18-transcript panel. Complementary bioinformatic analyses were performed solely to provide exploratory biological and clinical context for the experimentally prioritised transcripts. Propofol produced a concentration-dependent reduction in normalised MTT metabolic activity, with an MTT-derived IC₅₀ of 7.6 µM. At this concentration, total apoptosis increased from 0.67 ± 0.11% in vehicle-treated cells to 17.04 ± 2.40% following propofol exposure (p < 0.001), whereas necrosis remained minimal. The viable-cell fraction decreased from 99.30 ± 1.057% to 82.80 ± 0.841% (p < 0.05). The difference between the MTT-derived IC₅₀ and the Annexin V/7-AAD viable-cell fraction indicates that the MTT-derived IC₅₀ reflects reduced metabolic activity rather than 50% cell lethality. RT-qPCR showed decreased WNT5B, FZD5, and WNT11 transcript abundance and increased GSK3B transcript abundance. All four remained significant after Benjamini–Hochberg correction (WNT5B, q = 0.001; FZD5, q = 0.001; WNT11, q = 0.027; GSK3B, q = 0.041). Exploratory bioinformatic analyses showed heterogeneous gene-specific expression, survival, genomic, immune-infiltration, and dependency patterns and were not interpreted as evidence of a propofol-mediated mechanism in patients. Under the tested in vitro conditions, propofol reduced MTT-derived metabolic activity, increased apoptosis, and was associated with selective Wnt–Frizzled-related transcript changes in MIA PaCa-2 cells. These transcript-level observations do not establish direct functional modulation of Wnt–Frizzled signalling or demonstrate that this pathway mediates the observed apoptotic response. The bioinformatic findings provide exploratory contextual information only. Further validation in additional PDAC and non-malignant pancreatic cell models, together with protein-, phosphorylation-, and pathway-level functional studies, is required.

Objective: The pre-analytical phase is the stage at which laboratory errors most frequently occur and is therefore of critical importance for patient safety. During the COVID-19 pandemic, the marked increase in workload, organizational restructuring, staff and resource redeployment, changes in patient volume and clinical workflows, and disease-related pathophysiological alterations affecting specimen characteristics created unprecedented conditions that could influence laboratory quality indicators. This study aimed to characterize changes in specimen rejection rates across the pre-pandemic, pandemic, and normalization periods between 2018 and 2023, and to determine whether pandemic-related disruptions in health care delivery were associated with differences in the number, rate, and distribution of specimen rejections according to clinical unit, rejection reason, and time interval, with emphasis on the magnitude of these differences rather than statistical significance alone. The study also sought to generate evidence that could inform the development of preventive systems and strengthen laboratory preparedness for future large-scale disruptions to health care services. Materials and Methods: This retrospective observational study used routinely collected, aggregated laboratory information system (LIS) data from the Central Laboratory of Istanbul Atlas University Hospital for 1 January 2018 through 31 December 2023. The LIS extract was restricted to eligible specimen types, including spot and 24 h urine, serum, and whole-blood or plasma specimens collected in EDTA-, citrate-, or heparin-containing tubes; all other specimen types were excluded. Specimen rejection rates were evaluated by study period, clinical unit, time interval, and rejection reason. For cross-period comparisons, the total number of specimens within each respective study period was used as the denominator for calculating rejection rates, whereas for rejection-reason-specific analyses, the number of specimen types relevant to each rejection category was used as the denominator. Negative-binomial regression with denominator-based offsets estimated adjusted rejection-rate ratios (RRs) with 95% confidence intervals (CIs), with Bonferroni adjustment for selected pairwise comparisons. No missing or unclassifiable values were identified; no imputation was required. Reporting follows STROBE and the RECORD extension for studies using routinely collected health data. Results: The overall rejection rate increased progressively from 0.23% in the pre-pandemic period to 0.49% during the pandemic and 0.65% in the normalization period, and monthly observed rates supported this progressive increase. In adjusted terms, the rejection rate was 2.20-fold higher during the pandemic than in the pre-pandemic period (95% CI: 1.96–2.46) and increased a further 1.42-fold in the normalization period compared with the pandemic period (95% CI: 1.27–1.59). Across clinical units, the emergency department had the highest observed rejection rate, whereas intensive care units showed the highest adjusted rejection rates during the pandemic (RR = 1.34 versus the emergency department, 95% CI: 1.00–1.80) and normalization periods. Hemolysis and clotting were the dominant rejection reasons; although clotting had the highest observed rates in the pre-pandemic and pandemic periods, hemolysis ranked highest after adjustment in all periods and separated significantly from clotting only in the normalization period (RR = 1.97, 95% CI: 1.65–2.35). Time-interval effects were context-dependent rather than uniform and became informative mainly in combination with clinical unit or rejection reason: intensive care unit rejections peaked in the daytime interval during the pandemic (RR = 1.84 versus the evening interval, 95% CI: 1.06–3.19), and in the normalization period emergency department–hemolysis emerged as the leading adjusted clinical unit–rejection reason combination (RR = 1.89 versus intensive care unit–hemolysis, 95% CI: 1.19–3.00), while clotting remained centered on intensive care units and became more concentrated in the overnight and early-morning interval. Conclusions: Pre-analytical specimen rejection increased progressively across the pre-pandemic, pandemic, and normalization periods, with adjusted effect sizes indicating a substantial and sustained deterioration rather than a transient pandemic-related disturbance, and with persistent differences in rejection patterns according to clinical unit, time interval, and rejection reason. The further increase observed in the normalization period suggests that pandemic-related disruptions were associated with sustained changes in pre-analytical performance but were unlikely to be the sole explanation. Emergency department–hemolysis during the normalization period and intensive care unit–related clotting emerged as the most concrete quality-monitoring targets. Integrating rejection rates and counts with stratified monitoring by clinical unit, time interval, and rejection reason in combination may support earlier identification of emerging risks, more targeted preventive interventions, and stronger laboratory preparedness for future large-scale disruptions to health care services.