Hoshi University

Reactive oxygen species and reactive nitrogen species play roles in the pathogenesis of numerous diseases, but their presence is often assessed indirectly via analysis of specific biomarkers. Tyrosine-derived compounds such as ortho-tyrosine, meta-tyrosine, 3-chlorotyrosine, and 3-nitrotyrosine are considered markers of oxidative and nitrosative stress. However, these biomarkers are typically present in extremely low concentrations (in the low nanomolar range) and have structural isomers, making accurate quantification challenging. The aim of this study was to improve the sensitivity and chromatographic separation of these analytes using various commercially available derivatization reagents targeting amino groups. Selected reaction monitoring transitions were optimized by flow injection analysis of derivatized standards, and structural isomers were evaluated using LC-MS/MS. A Box-Behnken design and response surface methodology were employed to determine the optimal derivatization conditions for each reagent. Among the reagents tested, diethyl ethoxymethylenemalonate was most effective, enabling complete isomer separation and approximately 1000-fold signal enhancement compared with non-derivatized analytes, with a limit of quantification reaching 5 nM. The results of this study highlight the importance of selecting an appropriate derivatization strategy for sensitive and selective quantification of tyrosine-based biomarkers and offer a promising approach for the accurate assessment of oxidative and nitrosative stress in biological samples.

In Japan, human papillomavirus (HPV) vaccination rates remain low, partially owing to safety concerns from media reports. Pharmacies, as accessible healthcare facilities, often serve as sources of health information. However, the impact of passive information provision in pharmacies on public awareness and behavioral intentions has not been completely evaluated.

This study examined whether passive dissemination of HPV vaccine-related information in community pharmacies-through posters and leaflets-could improve visitors' knowledge and motivation to recommend HPV vaccinations.

A cluster randomized controlled trial was conducted in 20 pharmacies, of which 10 assigned were to the intervention group and 10 to the control group. Posters and leaflets on HPV vaccinations were displayed in the intervention group only, without the pharmacist's explanation. Adult visitors (n = 182) aged 20-75 years completed baseline and follow-up questionnaires using the vaccinaTion & Hpv Knowledge scale (THinK) and 7C Vaccination Readiness Scale. The primary endpoint was the change in attitudes toward HPV vaccination (aHPV)-a composite indicator reflecting willingness to receive the vaccine, recommend it to others, and seek related information-at 12 weeks.

No significant difference in aHPV score changes was observed between the groups at 12 weeks (mean difference = 0.14; p = 0.18). However, knowledge of HPV infection (kHPV) was significantly improved in the intervention group at 8 and 12 weeks (p < 0.05). No significant changes were observed in any of the subscales of the 7C Vaccination Readiness Scale.

Passive information provision in pharmacies improved HPV-related knowledge but did not enhance motivation to recommend vaccination. To enhance promotion, other active modalities-such as dialogue-based communication by professionals-may be more effective.

Cancer survivors are increasingly reported to exhibit signs of accelerated aging, largely attributed to the cytotoxic effects of chemotherapy, which may lead to various age-related conditions. Despite this, treatment-induced alterations in physical appearance-particularly changes in skin structure-are often overlooked in clinical practice and remain poorly understood. This study aimed to elucidate the mechanisms by which cytotoxic anticancer drugs affect skin integrity, with a focus on collagen (type I and III) and elastin, key components associated with skin aging. In a murine model, dietary restriction alone, as well as treatment with each of the anticancer drugs used in this study (cisplatin, 5-fluorouracil, vincristine, irinotecan and cyclophosphamide), led to significant weight loss; however, dermal thinning was observed exclusively in the cisplatin and vincristine. This structural deterioration correlated with a pronounced downregulation of Col1a1, Col1a2, Col3a1, and Eln at both the mRNA and protein levels. Mechanistically, this was accompanied by suppression of the TGF-β/Smad signaling pathway, evidenced by reduced TGF-β expression and Smad2 phosphorylation. Furthermore, the gene expression of Loxl1 and Loxl2-enzymes critical for collagen and elastin cross-linking was significantly diminished. These findings suggest that cisplatin and vincristine compromise dermal architecture by disrupting TGF-β signaling and extracellular matrix homeostasis, potentially contributing to premature skin aging in cancer survivors.