Background/Objectives: Oseltamivir is a widely used antiviral agent indicated for the treatment of influenza and plays a central role in pandemic preparedness strategies. Although an extemporaneously prepared 15 mg/mL oral solution is recognized in formularies, preparing large volumes during a pandemic requires extended storage options. However, no stability studies are currently available for an oseltamivir 15 mg/mL oral solution stored under freezing and subsequent refrigerated conditions for the duration of a standard treatment. The aim of this study was to evaluate the physicochemical and microbiological stability of a 15 mg/mL oseltamivir oral solution prepared from the active pharmaceutical ingredient (API) and packaged in amber glass containers. Methods: The oral solution was formulated using oseltamivir phosphate API, sodium benzoate as a preservative, and purified water and then packaged in 125 mL Type II amber glass bottles, allowing for thermal expansion. The samples were stored at -20 ± 2 °C for up to 75 days, followed by refrigerated storage (5 ± 3 °C) after thawing for up to 10 days. Chemical stability was assessed using a validated HPLC method in accordance with ICH guidelines and was defined as 90-110% recovery of the initial concentration. Physical stability (color, pH, particulate matter, crystallization, and homogeneity) and microbiological stability were also evaluated. Results: The HPLC method demonstrated excellent linearity, precision, and accuracy. Oseltamivir concentrations remained within the predefined acceptance limits throughout the 75-day study period under freezing conditions, with no significant changes in pH, color, or particulate formation. After thawing, the drug concentration continued to remain fully stable and within the required limits for up to 10 days under refrigerated conditions. Despite the prolonged storage and phase changes, no significant changes in physical parameters were observed, and microbiological testing confirmed the absence of aerobic, anaerobic, and fungal microorganisms on the final day of the study. Conclusions: Oseltamivir 15 mg/mL oral solution in amber glass bottles is physicochemically and microbiologically stable for up to 85 days (75 days under frozen conditions plus 10 days under refrigeration after thawing).

This study introduces "amber propolis," a unique variety from the Pampa biome in southern Brazil, characterized by a distinct chemical fingerprint and high biological potency. The composition was comprehensively analyzed via GC-MS and LC-MS/MS; biological activity was evaluated through antimicrobial (Staphylococcus aureus and Escherichia coli) and antileukemic screenings against K562, Jurkat, and U937 cell lines. Results revealed a terpenoid-dominant profile (comprising 60-70% of identified features) despite a lower flavonoid abundance compared to traditional reference samples. Amber propolis exhibited significantly higher antibacterial activity against S. aureus than green propolis, whereas all varieties showed similar dose-dependent inhibition of E. coli, reaching complete growth suppression at 500 µg/mL. Notably, it acted as a potent antileukemic agent, inducing rapid apoptosis starting at 12 h in Jurkat and U937 cell lines, and at 24 h for K562 cells, while concomitantly triggering G2/M cell cycle arrest. Unlike the variable responses observed for green and red propolis across lineages, amber propolis demonstrated a uniquely uniform cytotoxic effect, suggesting the involvement of distinct, conserved molecular mechanisms. These findings challenge the prevailing paradigm associating propolis bioactivity primarily with phenolic abundance. Collectively, this study establishes amber propolis as a chemically distinct and biologically active variety with strong potential for pharmaceutical and nutraceutical applications, supporting its future valorization and geographical indication within the Brazilian Pampa biome.

Blue and green fluorescent resins from the La Cumbre deposit in the Dominican Republic represent one of the most distinctive and sought-after varieties of amber, yet the nature of their unusual visible-light-induced luminescence remains insufficiently understood. In this study, we combine Ultraviolet-Visible (UV-Vis) Spectroscopy, Fluorescence Spectroscopy (including EM-EX matrices and lifetime measurements), Fourier-Transform Infrared Spectroscopy (FTIR), and Raman Spectroscopy (RS) - supported by hierarchical cluster analysis (HCA) and principal component analysis (PCA) - to characterize yellow (YC), red (RC), blue (BC), and green (GC) resins from the La Cumbre mine. UV-Vis data confirm that all samples exhibit absorption features consistent with a yellow to red body colour, whereas blue to blue-green fluorescence is observed most prominently under surface excitation and collection conditions. Fluorescence matrices of BC and GC resins reveal excitation-emission pairs consistent with one dominant fluorophore or fluorophore family in blue amber and a more complex set of emitting species in green resin. FTIR and Raman spectra show only subtle chemical differences among resin types; however, selected chemometric analyses highlighted YC/BC and RC/GC affinities, whereas full-range FTIR-HCA reflected broader matrix-level variability. In turn the shift from yellow to blue, or red-to-green is most plausibly explained by localized chemical and microstructural modifications occurring within closely related La Cumbre resin material, which aligns with their strong spectral similarities observed in the fluorescence, UV-Vis, RS and FTIR chemometric datasets. Our results suggest that the characteristic luminescence of La Cumbre amber arises from the combined influence of localized molecular and microstructural modifications developed during diagenesis and polymerization, rather than from the uniform distribution of a single fluorophore. Overall, the integrated multi-method spectroscopic approach, with fluorescence spectroscopy providing the clearest differentiation, offers a framework for characterising the investigated blue-fluorescent Dominican amber and its relationships to the other La Cumbre resin varieties.

Blue-light filters now span spectacle, contact and intraocular lenses, but the term encompasses technologies with markedly different spectral characteristics and intended outcomes. This state-of-the-art narrative review synthesises peer-reviewed evidence on optical performance, digital eye strain, glare, photostress, sleep and circadian physiology, retinal phototoxicity, macular health, reported adverse effects and commercial translation. Near-clear spectacle filters preserve high-contrast acuity in most studies, but randomised and systematic reviews show no clinically meaningful average reduction in short-term digital eye strain. Some products reduce blue-colour contrast, particularly under low-contrast or low-light conditions, and adverse-event reporting is sparse. Studies of evening filters are heterogeneous: substantial amber attenuation can alter circadian light exposure, yet several trials have found no objective sleep benefit. Conventional visual outcomes with blue-filtering intraocular lenses are broadly comparable with ultraviolet-only lenses, although some individual studies report small mesopic colour or contrast disadvantages; long-term macular protection remains unresolved. Research on contact lenses is limited, typically short-term and often manufacturer-associated, and focuses on glare, photostress, scatter and perceived brightness rather than long-term health outcomes. Taken together, the evidence should be interpreted by filter spectrum and intended outcome, rather than treating blue-light filtering as a single intervention.

A major challenge in genomics is deciphering the functional consequences of non-coding genetic variation. Here we present AlphaGenome Atlas, a comprehensive resource that enables the joint interpretation and prioritization of variant effects across the entire human genome. Using AlphaGenome, we predicted the regulatory effects across thousands of molecular phenotypes for every possible human single nucleotide variant and many observed indels. These predictions were then used to derive a unified and interpretable AlphaGenome Variant Impact (AVI) score and to map cis-regulatory motifs across the genome. AVI achieved state-of-the-art performance across diverse benchmarks with improved prioritization of deleterious non-coding variants. Application of the combined Atlas resource helped solve an epileptic encephalopathy rare disease case, increased the statistical power to detect rare non-coding variants driving population-level phenotypes, and enhanced the mechanistic interpretation of these variants. Thus, AlphaGenome Atlas improves the prioritization and molecular interpretation of non-coding variants with genetic and clinical significance.

Although pediatric pulled elbow (PE) typically responds well to standard closed reduction maneuvers, occasional irreducible episodes present a clinical challenge. Currently, optimal management guidelines for these refractory cases remain unestablished, particularly regarding the necessity of immediate surgical intervention or rigid immobilizatiom. We report the outcomes of a "supervised neglect" strategy-observation without external immobilization-for irreducible PE. We retrospectively reviewed six consecutive cases of irreducible PE treated at our institution between 2020 and 2025, and the mean age was 3.4 years. All patients were diagnosed with ultrasonography after failed manual reduction attempts. Patients were managed without external immobilization and monitored using ultrasonography until reduction was confirmed and supinator muscle swelling resolved. The median time from injury to the return of spontaneous arm movement was 4 days. The median time to ultrasonographic confirmation of reduction was 10 days, and the resolution of supinator swelling was 7 days postreduction. All cases resolved spontaneously without external immobilization or surgery. Interestingly functional arm movement returned before the sonographic confirmation of reduction. Furthermore, no residual symptoms or complications were observed in any case. These findings indicate that irreducible PE can be managed successfully by using a supervised neglect strategy. Spontaneous reduction occurred in all cases within 2 weeks; therefore, external immobilization and immediate surgery may not be necessary. Ultrasonography has proven to be a valuable tool for the diagnosis and monitoring of the healing process.

This perspective paper illustrates how model-informed vaccine development (MIVD) impacts vaccine discovery and development decisions, saving time and resources. It discusses the benefits of mechanistic and other mathematical models within MIVD. We outline key applications, opportunities, challenges, and future steps for MIVD as a call for the continued integration of mathematical modelling during vaccine development.

Squalene-based nanoemulsions are widely used as adjuvants in vaccine formulations, but their stability can be affected by environmental factors such as pH, oxidative stress, temperature, and light. We have produced a squalene nanoemulsion (CTVad1) to support the clinical development of new vaccines. This study aimed to develop and validate an HPLC method for squalene quantification in SpiN-Tec, a recombinant protein vaccine against COVID-19. We also aimed to evaluate the stability of the CTVad1 adjuvant and SpiN-Tec under controlled storage conditions. A reversed-phase HPLC method was developed and validated, and comprehensive forced degradation studies were performed on the raw material and SpiN-Tec under acidic, basic, oxidative, thermal, and photolytic conditions to demonstrate the stability-indicating capability of the method. Physicochemical, morphological, and biological characteristics were assessed, and stability studies of both the vaccine and the CTVad1 adjuvant were performed under accelerated and long-term conditions. The HPLC method was selective, precise, accurate, linear, and robust. Squalene raw material degraded under all tested conditions, whereas formulation in nano-sized globules improved its stability, with degradation observed only under hydrogen peroxide and light exposure. CTVad1 remained stable over time, exhibiting only minor, non-critical changes within the specification limits in both accelerated and long-term stability studies, regardless of the glass packaging used (clear or amber). In addition, the SpiN-Tec vaccine maintained its physicochemical and biological integrity under all tested conditions, with all evaluated parameters remaining within the established specification ranges. Our findings demonstrate that proper formulation, packaging, and storage conditions can preserve squalene stability in nanoemulsion-based vaccines, ensuring the quality, safety, and efficacy of the SpiN-Tec vaccine and its adjuvant throughout shelf life.

Psychiatric comorbidities are common in chronic pain and can meaningfully influence safety, satisfaction, adherence, and durability of benefit after interventional pain procedures. Interventional pain physicians, typically trained in anesthesiology, often have limited time and limited access to liaison psychiatry, creating a gap between guideline intent and real-world feasibility. This narrative review summarizes clinically relevant psychiatric domains for interventional pain practice, highlights situations where procedures should proceed, be optimized in parallel, or be deferred, and outlines a pragmatic approach to screening and first-line management in busy clinics. The authors describe brief screening tools, including validated instruments where applicable, that can be implemented within routine workflow to identify depression, anxiety, suicidality, substance misuse risk, and maladaptive coping, and they propose the Daradia Protocol, a structured 10-minute stepped-care pathway that stratifies patients into Green, Amber, and Red risk categories to guide procedural decision-making and referral urgency. Clinic-initiated strategies emphasize psychoeducation, expectation alignment, graded activity, sleep support, and coordinated care with mental health professionals when indicated. Embedding rapid screening with clear action thresholds may strengthen informed consent, improve patient selection, and increase the likelihood of durable outcomes without materially extending consultation time.

Traditional deep mutational scanning (DMS) encodes every single amino-acid substitution from a wild-type sequence. We hypothesize that combinatorial DMS (CDMS) libraries, incorporating all mutations in all combinations, can enable the discovery of high-affinity protein (super)binders by capturing epistatic, non-linear amino-acid interactions. Here, we introduce origin-independent and context-exhaustive high-throughput integration of combinatorial DMS libraries (ORCHID), which systematically maps regions of wild-type-independent epistasis across all mutational contexts and trajectories. For benchmarking, we build a high-throughput peptide display assay measuring PIN1[WW-domain] affinity for a CDMS peptide library containing phosphoserine via amber codon suppression. ORCHID raises prediction accuracy by 45% over non-epistatic models. We identify and structurally characterize two epistatic superbinders, SPY-tide and LYR-tide, binding 3- to 5-fold tighter than current optimal PIN1[WW-domain] binders through "fold-and-turn" and register-shifted conformational changes. We also identify two molecular determinants of epistasis, PIN1[F25] and PIN1[R14], that natively encode non-linear binding and, when mutated, abolish it. Hence, natural proteins recognize peptides non-linearly, offering opportunities for improved binder design.

Pseudobulbar affect is a distressing condition frequently observed in patients with amyotrophic lateral sclerosis (ALS), for which a fixed-dose combination of dextromethorphan and quinidine is approved in the United States. In Europe, no commercial product is available, and compounded capsules or suspensions are used; however, capsules may be unsuitable for patients with dysphagia, and suspensions require shaking before administration. The aim of this study was to develop simple fully water-soluble formulation of dextromethorphan hydrobromide and quinidine sulfate using methyl-β-cyclodextrin as a solubilizing agent and to evaluate its stability under different storage conditions, enabling pharmacies to compound the preparation without complex infrastructure. Preliminary qualitative solubility experiments showed that the dextromethorphan hydrobromide/ quinidine sulfate powder mixture did not dissolve in water without methyl-β-cyclodextrin. Addition of methyl-β-cyclodextrin improved dissolution; however, the initial preparation targeting 20 mg/mL dextromethorphan hydrobromide and 10 mg/mL quinidine sulfate in a final volume of 100 mL remained incompletely dissolved. Increasing the final volume to 250 mL yielded a clear formulation containing nominal concentrations of 8.4 mg/mL dextromethorphan hydrobromide and 4.2 mg/mL quinidine sulfate. Three independently prepared batches were stored in amber PET bottles providing light protection and fitted with child-resistant closure at 2-8°C, 15-25°C, and 40°C and analyzed by HPLC for four weeks. HPLC assay values remained within 10% of the corresponding baseline values at each storage condition, and no visible precipitation was observed. The results support the physical clarity and chemical consistency of the tested methyl-β-cyclodextrin- containing formulation over four weeks under the evaluated storage conditions. Microbiological stability, preservative effectiveness, formal method validation in the author's laboratory, and the extent of cyclodextrin inclusion complexation were not assessed. This formulation may serve as a basis for further pharmaceutical, microbiological, and clinical evaluation of an extemporaneously compounded oral liquid.

Coat protein (CP) tertiary structures and their capsid organization of spherical viruses are generally well conserved within each virus family. While AlphaFold successfully predicts the tertiary structures of individual CPs, their association to form proper quaternary assemblies cannot be easily accomplished. Here, we report a generalized methodology and an associated web-based utility (https://foldavirus.org) that combines AlphaFold predictions of CPs with the knowledge on corresponding icosahedral architectures (e.g., T = 1, 3, 4…) based on the known structures from the related virus family to generate associated capsids. The resulting assemblies are relaxed using Amber energy minimization to relieve any steric clashes at the intersubunit interfaces. Significantly, the capsid models are validated by calculating robust Mahalanobis distance using the residue annotations categorized as interface, core, and surface amino acids with respect to those observed in the experimentally determined structures from the corresponding virus family. Given the amino acid sequence of CP(s), we successfully generated capsids up to T = 9 icosahedral symmetry, including those of picornaviruses that display pseudo-T = 3 symmetry comprising VP1-VP4. As the number of currently available CP sequences are 2 to 3 orders of magnitude larger than the experimentally determined 3D-structures, this approach bridges the huge gap that exists between the sequence and structural space of viruses.