Inonotus obliquus, a medicinal mushroom valued for its bioactive compounds, has not been previously characterized from Romanian sources. This study presents the first comprehensive chemical and biological screening of I. obliquus, introducing novel polymer-based encapsulation systems to enhance the stability and bioavailability of its bioactive constituents. Two distinct delivery systems were designed to enhance the functionality of I. obliquus extracts: (i) microencapsulation in maltodextrin (MIO) and (ii) a sequential approach involving preparation of silver nanoparticle-loaded I. obliquus (IO–AgNPs), followed by microencapsulation to yield the hybrid MIO–AgNP system. Comprehensive metabolite profiling using GC–MS and ESI–QTOF–MS revealed 142 bioactive constituents, including terpenoids, flavonoids, phenolic acids, amino acids, coumarins, styrylpyrones, fatty acids, and phytosterols. Structural integrity and successful encapsulation were confirmed by XRD, FTIR, and SEM analyses. Both IO–AgNPs and MIO–AgNPs demonstrated potent antioxidant activity, significant acetylcholinesterase inhibition, and robust antimicrobial effects against Staphylococcus aureus, Bacillus cereus, Pseudomonas aeruginosa, and Escherichia coli. Cytotoxicity assays revealed pronounced activity against MCF-7, HCT116, and HeLa cell lines, with MIO–AgNPs exhibiting superior efficacy. The synergistic integration of maltodextrin and AgNPs enhanced compound stability and bioactivity. As the first report on Romanian I. obliquus, this study highlights its therapeutic potential and establishes polymer-based nanoencapsulation as an effective strategy for optimizing its applications in combating microbial resistance and cancer.

Background and Objectives: p16 immunohistochemistry (IHC) is widely used as a surrogate marker for high-risk human papillomavirus (HPV)-driven carcinogenesis in oropharyngeal squamous cell carcinoma. Diagnostic reliability in laryngeal squamous cell carcinoma (LSCC) is still up for debate, particularly because HPV DNA detection, p16 overexpression, and transcriptionally active HPV infection may be discordant at this anatomical site. This systematic review aimed to assess the diagnostic performance of p16 IHC as a surrogate marker for high-risk HPV status in primary invasive LSCC. Materials and Methods: A systematic review was conducted in accordance with PRISMA principles and prospectively registered in PROSPERO. PubMed, Scopus, and Web of Science, and citation searches were used to identify studies reporting paired p16 IHC and tumour-based molecular HPV testing in invasive LSCC. Eligible HPV reference standards included HPV DNA PCR/genotyping, DNA in situ hybridization, RNA in situ hybridization, and E6/E7 mRNA detection. Data were synthesized narratively because of substantial heterogeneity in p16 thresholds, HPV assays, and study populations. Results: Fourteen studies were included. p16 positivity thresholds varied widely, ranging from ≥30% moderate/strong staining to ≥70–75% diffuse nuclear and cytoplasmic staining. HPV reference standards also differed substantially across studies. Overall, p16 IHC showed inconsistent concordance with molecular HPV testing. Studies using RNA-based reference standards showed that transcriptionally active HPV was uncommon in LSCC and that p16-positive tumours often lacked evidence of active HPV transcription. Conclusions: p16 IHC should not be used as a standalone surrogate marker for high-risk HPV-driven carcinogenesis in invasive LSCC. When HPV attribution is clinically or analytically important, p16-positive cases should be confirmed using HPV-specific molecular testing, preferably RNA-based assays. Future prospective studies using standardized p16 protocols and transcriptionally active HPV reference standards are needed.

Background and Objectives: Vitamin D deficiency and psychological distress have been linked to cancer biology, but their relevance to tumor differentiation in head-and-neck squamous cell carcinoma (HNSCC) is uncertain. Materials and Methods: In this cross-sectional study at the Department of Otolaryngology, County Hospital of Timișoara, Romania, we enrolled newly diagnosed HNC patients from October 2023 to December 2024, analyzing 199 SCC patients after exclusions. Vitamin D status was assessed using serum 25-OH-vitamin D levels, and distress was measured with the validated Romanian version of the Hospital Anxiety and Depression Scale (HADS). Tumor aggressiveness was defined by histological grade (G3 vs. G1–G2). Univariate, multivariate, and subgroup analyses were conducted, adjusting for confounders like smoking. Results: Vitamin D deficiency (<20 ng/mL) was prevalent (80.40%), with median 25-OH-vitamin D levels of 15.1 ng/mL. Univariate analysis revealed a modest association between vitamin D deficiency and poorly differentiated tumors (G3 vs. G1–G2; OR = 1.79, p = 0.055) and between clinically significant anxiety (HADS-A ≥ 8) and G3 tumors (OR = 1.71, p = 0.059). A weak negative correlation was observed between 25-OH-vitamin D levels and HADS-A scores (rho = −0.17, p = 0.052). In multivariate analysis adjusted for age, smoking, and tumor location, these associations weakened (vitamin D deficiency: OR = 1.55, p = 0.082; HADS-A ≥8: OR = 1.56, p = 0.113). Subgroup analysis suggested a trend toward higher odds of G3 tumors in patients with both vitamin D deficiency and high anxiety (OR = 1.72, p = 0.075). Conclusions: Univariate analyses indicated potential links between vitamin D deficiency, psychological distress, and tumor aggressiveness in HNSCC, but these did not reach statistical significance after adjustment for confounders. The observed trends, particularly in subgroups with combined deficiency and distress, suggest a possible interplay worth exploring further. To conclude, neither vitamin-D deficiency nor clinically significant distress independently predicted poor histological differentiation after adjustment; observed trends, including a possible distress–vitamin-D interaction, are hypothesis-generating and warrant testing in larger, longitudinal cohorts.