Indiana University Health

High-throughput transcriptomic technologies have advanced rapidly, enabling genome-wide gene expression profiling. Microarrays, introduced in 1995, laid the foundation for large-scale analysis but were later surpassed in 2008 by RNA sequencing (RNA-seq), which offers single-nucleotide resolution, detects low-abundance transcripts, and does not require prior sequence knowledge. Bulk RNA-seq provides robust insights into global transcriptomic changes but lacks single-cell resolution. Single-cell RNA-seq (scRNA-seq), introduced in 2009, addressed this limitation by revealing cellular heterogeneity and dynamic gene expression. However, its application in bone research is constrained due to difficulties in releasing bone cells called osteocytes from the mineralized matrix, often resulting in low yield and dissociation-induced artifacts. In order to address these challenges, single-nucleus RNA-seq (snRNA-seq), first introduced in 2016 to enable transcriptomic profiling from isolated nuclei, was used in this study. We developed a protocol for snRNA-seq on bone tissue, achieving high-yield recovery of osteocyte nuclei from snap-frozen, marrow-flushed long bones. This approach minimized dissociation bias and enhanced osteocyte representation. We applied this robust method to long bones from young adult male and female mice, generating a high-resolution map of osteocyte gene expression under physiological conditions. Compared to scRNA-seq datasets, where osteocytes represent only 0.18%-6.64% of cells, our snRNA-seq approach increased osteocyte capture and transcriptomic fidelity to 18.5%. We identified an osteocyte transcriptomic signature highlighting the top 30 genes, including Sost, which is typically undetected or lowly-expressed in scRNA-seq. Notably, 23 of these genes have not been well-characterized in osteocytes, including Tg, Kcnq5, Rapgef4os1, Cacna1a, Egr3, Dok5, and Lgr6, which may represent novel regulators of osteocyte biology. This study represents the first application of snRNA-seq specifically for osteocyte analysis in bone tissue, providing a valuable resource for investigating osteocyte biology and skeletal disorders.

Sport-related head exposure characteristics (i.e., concussion history, sport contact classification, and total years of sport participation) are suggested to be associated with short-term and long-term health outcomes in competitive athletes. However, their relationship with intermediate health outcomes (specifically within 5 years of collegiate sport retirement) remains unknown. Therefore, we examined associations between sport-related head exposure characteristics and physical, mental, cognitive, and behavioral health measures among former collegiate athletes. Former collegiate athletes who completed a baseline evaluation between 2018 and 2021 and were evaluated within 5 years of collegiate graduation were included. Primary predictors included lifetime concussion history (0 [referent], 1-2, 3+), sport contact classification (unexposed [ref.], low, high), and total years of sport participation (continuous). Outcome measures collected included the Alcohol Use Disorders Identification Test, Brief Symptom Inventory-18 (BSI-18), Neuro-Quality of Life cognitive domain (Neuro-QoL), Patient Health Questionnaire (PHQ-9), Pittsburgh Sleep Quality Index (PSQI), Sport Concussion Assessment Tool (SCAT) symptom inventory, 12-Item Short Form Survey (SF-12), and Satisfaction with Life Scale. Twelve multivariable linear regression models with bootstrapping were constructed, with adjusted unstandardized beta coefficients (BAdj) and standard errors (SEs) estimated. Overall, 3,910 former collegiate student-athletes were examined; 48.6% were female, 73.0% attended a Division I school, 36.2% self-reported ≥1 lifetime diagnosed concussion, 38.1% participated in a high-exposure sport, and the median years since college graduation to assessment was 5.0. Independently, scores of BSI anxiety (BAdj = 1.24, SE = 0.36), BSI depression (BAdj = 0.88, SE = 0.37), global severity index [GSI] (BAdj = 2.66, SE = 0.91), PHQ-9 (BAdj = 1.34, SE = 0.40), PSQI (BAdj = 0.69, SE = 0.29), SCAT total symptom severity (BAdj = 5.14, SE = 1.31), and SF-12 mental composite score (BAdj = -3.39, SE = 0.83) were worse in athletes with 3+ concussions compared with athletes without a history of concussion. Athletes with 1-2 concussions vs no concussion history similarly had worse scores independently for BSI-18 GSI (BAdj = 0.88, SE = 0.35), anxiety (BAdj = 0.40, SE = 0.15), Neuro-QoL (BAdj = -0.97, SE = 0.36), PHQ-9 (BAdj = 0.77, SE = 0.19), PSQI (BAdj = 0.27, SE = 0.13), SCAT total symptom severity (BAdj = 2.13, SE = 0.55), and SF-12 mental composite score (BAdj = -1.80, SE = 0.41). Lifetime concussion history was the most common predictor of adverse self-reported health outcomes compared with other exposure characteristics, within 5 years of collegiate sport retirement. Despite the associations, most former athletes remained within normal clinical levels during this early postretirement period.

To evaluate the impact of surface treatments and chemical denture cleansers on the surface color stability, gloss, and wettability of compression-molded, 3D-printed, and milled denture base materials. Specimens (12 × 16 × 7mm) were fabricated from a standardized STL file in three denture base materials (compression-molded, 3D-printed, and milled resins; n = 80 each) and assigned to polish-only or protective-coated (Optiglaze) subgroups. Specimens were immersed in one of four cleaning solutions: Polident (alkaline peroxide), Val-Clean (potassium caroate), Renew (alkaline hypochlorite), or distilled water (control group). Thirty immersion cycles per day were performed for 12 days, for a total of 360 cycles, simulating approximately 1 year of once‑daily denture cleaning. Color stability was evaluated using a spectrophotometer with the CIEDE2000 (ΔE00) formula. Surface gloss was measured with a gloss meter, and wettability was assessed by static contact angle using an optical tensiometer. Statistical analyses were conducted using three-way ANOVA. Color changes on acceptability and perceptibility were evaluated using one-sided, one-sample t-tests (thresholds of 1.8 and 0.8, α = 0.05). Renew produced the highest mean ΔE00 (3.90 ± 0.33), exceeding distilled water (1.76 ± 0.33), Polident (0.92 ± 0.80), and Val-Clean (0.78 ± 0.56). Using the acceptability threshold (AT) of ΔE00 = 1.8, all Polident and Val-Clean groups remained acceptable for milled and 3D-printed materials. In contrast, compression-molded materials exhibited greater color instability. All Renew groups exceeded the AT regardless of surface treatment or fabrication method. Gloss change was also significant; Optiglaze combined with Val-Clean caused some of the largest gloss losses in 3D‑printed samples (-17.99 ± 9.82, p < 0.0001) and compression-molded samples (-13.13 ± 4.21, p < 0.0001). Polished surfaces generally lost less gloss, such as Polident-treated compression-molded resin (-5.92 ± 3.33, p = 0.0048). For wettability, Optiglaze increased hydrophobicity in 3D-printed specimens in distilled water (+6.33 ± 7.11°, p = 0.0033) but decreased it when combined with Renew in compression-molded resin (-18.54 ± 6.70°, p < 0.0001). Polident or Val-Clean on polished surfaces best preserved color, gloss, and wettability. Renew produced the greatest overall changes in color and wettability. Gloss changes were strongly material‑ and treatment‑dependent. Optiglaze‑coated 3D‑printed and compression-molded bases immersed in Val-Clean showed pronounced gloss loss. Polished surfaces generally retained gloss better. Compression-molded materials were more susceptible to color change than milled or 3D-printed materials, and the Optiglaze did not consistently reduce changes in color, gloss, or wettability. These findings may inform future research and the development of evidence-based denture-cleaning protocols; however, clinical studies are needed before specific recommendations can be made.

Fluorine has been widely incorporated into small-molecule drugs as an effective strategy to modulate metabolic stability, lipophilicity, target interactions, and overall pharmacokinetic profile. As a result, fluorine-containing compounds remain well represented among newly approved therapeutics across a broad spectrum of disease areas; notably, in 2025, 14 of the 29 small-molecule drugs approved by the U.S. Food and Drug Administration (FDA) contained at least one fluorine atom, underscoring the continued relevance of fluorine in contemporary drug design. This annual review highlights the role of fluorine in modern medicinal chemistry by summarizing FDA-approved fluorine-containing small-molecule drugs in 2025. The review examines their therapeutic indications, molecular targets, and structural features, and outlines representative synthetic strategies used in their preparation. Overall, this review provides an updated perspective on the continued importance of fluorine chemistry in modern drug discovery and development.