Association between Preexisting Sleep Disorders and Oncologic Outcome in Patients with Oral Cavity Squamous Cell Carcinoma: A Nationwide Propensity Score-Matched Population-Based Cohort Study.

Purpose: To investigate the effects of preexisting sleep disorders on the oncologic outcomes of patients receiving standard treatments for oral squamous cell carcinoma (OSCC). Methods: The patients recruited from the Taiwan Cancer Registry Database who had received surgery for stage I−IVB OSCC. The Cox proportional hazards model was used to analyze all-cause mortality, locoregional recurrence (LRR), and distant metastasis (DM). The patients were categorized into those with and without sleep disorders (Groups 1 and 2, respectively) through propensity score matching. Results: In the multivariate Cox regression analysis, the adjusted hazard ratios for all-cause mortality, LRR, and DM for Group 1 compared with Group 2 were 1.19 (95% confidence interval (CI): 1.04−1.36; p = 0.011), 1.47 (95% CI: 1.23−1.75; p < 0.001), and 1.15 (95% CI: 1.02−1.44; p = 0.025), respectively. Conclusion: OSCC patients with sleep disorders demonstrated poorer oncologic outcomes than did those without sleep disorders. Therefore, before OSCC surgery, patients with OSCC should be screened for preexisting sleep disorders because they may serve as predictors for survival in these patients. Future studies investigating the survival benefits of pharmacological and behavioral treatments for sleep problems in patients with OSCC are warranted.

Emerging biomaterials for downstream manufacturing of therapeutic proteins.

Downstream processing is considered one of the most challenging phases of industrial manufacturing of therapeutic proteins, accounting for a large portion of the total production costs. The growing demand for therapeutic proteins in the biopharmaceutical market in addition to a significant rise in upstream titers have placed an increasing burden on the downstream purification process, which is often limited by high cost and insufficient capacities. To achieve efficient production and reduced costs, a variety of biomaterials have been exploited to improve the current techniques and also to develop superior alternatives. In this work, we discuss the significance of utilizing traditional biomaterials in downstream processing and review the recent progress in the development of new biomaterials for use in protein separation and purification. Several representative methods will be highlighted and discussed in detail, including affinity chromatography, non-affinity chromatography, membrane separations, magnetic separations, and precipitation/phase separations. STATEMENT OF SIGNIFICANCE: Nowadays, downstream processing of therapeutic proteins is facing great challenges created by the rapid increase of the market size and upstream titers, starving for significant improvements or innovations in current downstream unit operations. Biomaterials have been widely used in downstream manufacturing of proteins and efforts have been continuously devoted to developing more advanced biomaterials for the implementation of more efficient and economical purification methods. This review covers recent advances in the development and application of biomaterials specifically exploited for various chromatographic and non-chromatographic techniques, highlighting several promising alternative strategies.

Diagnosis of immunomarkers in vivo via multiplexed surface enhanced Raman spectroscopy with gold nanostars.

In this work, we demonstrate the targeted diagnosis of immunomarker programmed death ligand 1 (PD-L1) and simultaneous detection of epidermal growth factor receptor (EGFR) in breast cancer tumors in vivo using gold nanostars (AuNS) with multiplexed surface enhanced Raman spectroscopy (SERS). Real-time longitudinal tracking with SERS demonstrated maximum accumulation of AuNS occurred 6 h post intravenous (IV) delivery, enabling detection of both biomarkers simultaneously. Raman signal correlating to both PD-L1 and EGFR decreased by ∼30% in control tumors where receptors were pre-blocked prior to AuNS delivery, indicating both the sensitivity and specificity of SERS in distinguishing tumors with different levels of PD-L1 and EGFR expression. Our in vivo study was combined with the first demonstration of ex vivo SERS spatial maps of whole tumor lesions that provided both a qualitative and quantitative assessment of biomarker status with near cellular-level resolution. High resolution SERS maps also provided an overview of AuNS distribution in tumors which correlated well with the vascular density. Mass spectrometry showed AuNS accumulation in tumor and liver, and clearance via spleen, and electron microscopy revealed AuNS were endocytosed in tumors, Kupffer cells in the liver, and macrophages in the spleen. This study demonstrates that SERS-based diagnosis mediated by AuNS provides an accurate measure of multiple biomarkers both in vivo and ex vivo, which will ultimately enable a clinically-translatable platform for patient-tailored immunotherapies and combination treatments.

Bioinspired supramolecular engineering of self-assembling immunofibers for high affinity binding of immunoglobulin G.

Many one-dimensional (1D) nanostructures are constructed by self-assembly of peptides or peptide conjugates containing a short ß-sheet sequence as the core building motif essential for the intermolecular hydrogen bonding that promotes directional, anisotropic growth of the resultant assemblies. While this molecular engineering strategy has led to the successful production of a plethora of bioactive filamentous ß-sheet assemblies for interfacing with biomolecules and cells, concerns associated with effective presentation of α-helical epitopes and their function preservation have yet to be resolved. In this context, we report on the direct conjugation of the protein A mimicking peptide Z33, a motif containing two α-helices, to linear hydrocarbons to create self-assembling immuno-amphiphiles (IAs). Our results suggest that the resulting amphiphilic peptides can, despite lacking the essential ß-sheet segment, effectively associate under physiological conditions into supramolecular immunofibers (IFs) while preserving their native α-helical conformation. Isothermal titration calorimetry (ITC) measurements confirmed that these self-assembling immunofibers can bind to the human immunoglobulin G class 1 (IgG1) with high specificity at pH 7.4, but with significantly weakened binding at pH 2.8. We further demonstrated the accessibility of Z33 ligand in the immunofibers using transmission electron microscopy (TEM) and confocal imaging. We believe these results shed important light into the supramolecular engineering of α-helical peptides into filamentous assemblies that may possess an important potential for antibody isolation.

Conformation Preservation of α-Helical Peptides within Supramolecular Filamentous Assemblies.

Hydrogen-bonded ß-sheets are the most commonly explored building motifs for creating peptide-based filamentous nanostructures; however, most bioactive epitopes must assume an α-helix conformation to exert their functions. Incorporating α-helical sequences into ß-sheet-forming peptides often involves the use of a flexible spacer to alleviate the steric impact of the intermolecular hydrogen bonding on the α-helical conformation. In this context, we report our findings on the alkylation-regulated conformation preservation of α-helical peptides within their filamentous assemblies. We found that the chemical conjugation of two short linear hydrocarbons (octanoic acids, C8) can retain the α-helical conformation of two protein A-derived peptide sequences while effectively driving their assembly into filamentous nanostructures. In contrast, the use of a single palmitoyl tail (C16) of similar hydrophobicity would lead to formation of ß-sheet assemblies. Our studies further demonstrated that the length of the conjugated hydrocarbon also plays an important role in partially preserving the native α-helical conformation, with longer ones promoting ß-sheet formation and short ones stabilizing α-helices to some extent. We believe that these findings offer important guiding principles for the alkylation of self-assembling peptides containing α-helical sequences.