A Quantitative Re-Assessment of Microencapsulation in (Pre-Treated) Yeast.

Most hydrophobes easily diffuse into yeast cells, where they experience reduced evaporation and protection from oxidation, thus allowing inherently biocompatible encapsulation processes. Despite a long-standing industrial interest, the effect of parameters such as how is yeast pre-treated (extraction with ethanol, plasmolysis with hypertonic NaCl, depletion to cell walls), the polarity of the hydrophobes and the process conditions are still not fully understood. Here, we have developed thorough analytical protocols to assess how the effects of the above on S. cerevisiae's morphology, permeability, and encapsulation efficiency, using three differently polar hydrophobes (linalool, 1,6-dihydrocarvone, limonene) and three separate processes (hydrophobes as pure 'oils', water dispersions, or acetone solutions). The harsher the pre-treatment (depleted > plasmolyzed/extracted > untreated cells), the easier the diffusion into yeast became, and the lower both encapsulation efficiency and protection from evaporation, possibly due to denaturation/removal of lipid-associated (membrane) proteins. More hydrophobic terpenes performed worst in encapsulation as pure 'oils' or in water dispersion, but much less of a difference existed in acetone. This indicates the specific advantage of solvents/dispersants for 'difficult' compounds, which was confirmed by principal component analysis; furthering this concept, we have used combinations of hydrophobes (e.g., linalool and α-tocopherol), with one acting as solvent/enhancer for the other. Our results thus indicate advantages in using untreated yeast and-if necessary-processes based on solvents/secondary hydrophobes.

Early CAR- CD4+ T-lymphocytes recovery following CAR-T cell infusion: A worse outcome in diffuse large B cell lymphoma.

CAR- CD4+ T cell lymphopenia is an emerging issue following CAR-T cell therapy. We analyzed the determinants of CD4+ T cell recovery and a possible association with survival in 31 consecutive patients treated with commercial CAR-T for diffuse large B-cell (DLBCL) or mantle cell lymphoma. Circulating immune subpopulations were characterized through multiparametric-flow cytometry. Six-month cumulative incidence of CAR- CD4+ T cell recovery (≥200 cells/µL) was 0.43 (95% confidence interval [CI]: 0.28-0.65). Among possible determinants of CD4+ T cell recovery, we recognized infusion of a 4-1BB product (tisagenlecleucel, TSA) in comparison with a CD28 (axicabtagene/brexucabtagene, AXI/BRX) (hazard ratio [HR] [95% CI]: 5.79 [1.16-24.12] p = 0.016). Higher CD4+ T cell counts resulted with TSA at month-1, -2 and -3. Moderate-to-severe infections were registered with prolonged CD4+ T cell lymphopenia. Early, month-1 CD4+ T cell recovery was associated with a worse outcome in the DLBCL cohort, upheld in a multivariate regression model for overall survival (HR: 4.46 [95% CI: 1.12-17.71], p = 0.03). We conclude that a faster CAR- CD4+ T cell recovery is associated with TSA as compared to AXI/BRX. Month-1 CAR- CD4+ T cell subset recovery could represent a "red flag" for CAR-T cell therapy failure in DLBCL patients.

CD19-Targeted Immunotherapies for Diffuse Large B-Cell Lymphoma.

Surgical resection, chemotherapy and radiotherapy were, for many years, the only available cancer treatments. Recently, the use of immune checkpoint inhibitors and adoptive cell therapies has emerged as promising alternative. These cancer immunotherapies are aimed to support or harness the patient's immune system to recognize and destroy cancer cells. Preclinical and clinical studies, based on the use of T cells and more recently NK cells genetically modified with chimeric antigen receptors retargeting the adoptive cell therapy towards tumor cells, have already shown remarkable results. In this review, we outline the latest highlights and progress in immunotherapies for the treatment of Diffuse Large B-cell Lymphoma (DLBCL) patients, focusing on CD19-targeted immunotherapies. We also discuss current clinical trials and opportunities of using immunotherapies to treat DLBCL patients.

NK Cell-Based Immunotherapy in Colorectal Cancer.

Human Natural Killer (NK) cells are all round players in immunity thanks to their powerful and immediate response against transformed cells and the ability to modulate the subsequent adaptive immune response. The potential of immunotherapies based on NK cell involvement has been initially revealed in the hematological setting but has inspired the design of different immune tools to also be applied against solid tumors, including colorectal cancer (CRC). Indeed, despite cancer prevention screening plans, surgery, and chemotherapy strategies, CRC is one of the most widespread cancers and with the highest mortality rate. Therefore, further efficient and complementary immune-based therapies are in urgent need. In this review, we gathered the most recent advances in NK cell-based immunotherapies aimed at fighting CRC, in particular, the use of monoclonal antibodies targeting tumor-associated antigens (TAAs), immune checkpoint blockade, and adoptive NK cell therapy, including NK cells modified with chimeric antigen receptor (CAR-NK).

Transparent Bioplastic Derived from CO2-Based Polymer Functionalized with Oregano Waste Extract toward Active Food Packaging.

Active packaging materials, biodegradable and from renewable resources, are the most promising substitutes of nonbiodegradable, petroleum-based plastics, toward green and sustainable packaging solutions. In this study, an innovative bioplastic system, composed of carbon dioxide-derived poly(propylene carbonate) (PPC) and nature-originated cellulose acetate (CA), was developed. The extract from oregano waste was incorporated into the bioplastics as a low-cost and effective antioxidant resource. Thin, freestanding, and flexible PPC.CA bioplastic films were obtained by a simple, easily scalable solvent casting technique. The pristine films, without the oregano extract, featured good transparency and high water vapor barrier ability, along with suitable mechanical and thermal properties that are comparable to commercial plastics used for packaging. Interestingly, the incorporation of oregano waste extract added to the bioplastics high UV protection and high antioxidant activity, suitable features for active food packaging applications, without compromising the intriguing properties of the pristine films. The biocomposite films were not only biocompatible but also started biodegrading after just 1 week in seawater. The reported biocomposites are foreseen as promising candidates for several packaging applications, but in particular for sustainable active food packaging.

Role of the Main Non HLA-Specific Activating NK Receptors in Pancreatic, Colorectal and Gastric Tumors Surveillance.

Human NK cells can control tumor growth and metastatic spread thanks to their powerful cytolytic activity which relies on the expression of an array of activating receptors. Natural cytotoxicity receptors (NCRs) NKG2D and DNAM-1 are those non-HLA-specific activating NK receptors that are mainly involved in sensing tumor transformation by the recognition of different ligands, often stress-induced molecules, on the surface of cancer cells. Tumors display several mechanisms aimed at dampening/evading NK-mediated responses, a relevant fraction of which is based on the downregulation of the expression of activating receptors and/or their ligands. In this review, we summarize the role of the main non-HLA-specific activating NK receptors, NCRs, NKG2D and DNAM-1, in controlling tumor growth and metastatic spread in solid malignancies affecting the gastrointestinal tract with high incidence in the world population, i.e., pancreatic ductal adenocarcinoma (PDAC), colorectal cancer (CRC), and gastric cancer (GC), also describing the phenotypic and functional alterations induced on NK cells by their tumor microenvironment.

Investigation of in vitro hydrophilic and hydrophobic dual drug release from polymeric films produced by sodium alginate-MaterBi® drying emulsions.

Emulsions are known to be effective carriers of hydrophobic drugs, and particularly injectable emulsions have been successfully implemented for in vivo controlled drug release. Recently, high internal phase emulsions have also been used to produce porous polymeric templates for pharmaceutical applications. However, emulsions containing dissolved biopolymers both in the oil and water phases are very scarce. In this study, we demonstrate such an emulsion, in which the oil phase contains a hydrophobic biodegradable polymer, MaterBi®, and the water phase is aqueous sodium alginate dispersion. The two phases were emulsified simply by ultrasonic processing without any surfactants. The emulsions were stable for several days and were dried into composite solid films with varying MaterBi®/alginate fractions. The films were loaded with two model drugs, a hydrophilic eosin-based cutaneous antiseptic and the hydrophobic curcumin. Drug release capacity of the films was investigated in detail, and controlled release of each model drug was achieved either by tuning the polymer fraction in the films during emulsification or by crosslinking sodium alginate fraction of the films by calcium salt solution immersion. The emulsions can be formulated to carry either a single model drug or both drugs depending on the desired application. Films demonstrate excellent cell biocompatibility against human dermal fibroblast, adult cells.

Light Responsive Silk Nanofibers: An Optochemical Platform for Environmental Applications.

Photochromic spiropyran-doped silk fibroin poly(ethylene oxide) nanofibers which combine the attractive properties and biocompatibility of silk with the photocontrollable and reversible optical, mechanical, and chemical response of the spiropyran dopants are herein presented. As proved, the reversible variation of the absorption and emission signals of the mats and of their Young's modulus upon alternate UV and visible light irradiation is ascribed to the reversible photoconversion of the spiropyran form to its polar merocyanine counterpart. Most importantly, the interactions of the merocyanine molecules with acidic vapors as well as with heavy metal ions dispersed in solution produce analyte-specific spectral changes in the emission profile of the composite, accompanied by a characteristic chromic variation. Because of the high surface-to-volume ratio of the nanofibrous network, such interactions are fast, thus enabling both an optical and a visual detection in a 30-60 s time scale. The sensing platform can be easily regenerated for more than 20 and 3 cycles upon acid or ion depletion, respectively. Overall, the photocontrolled properties of the silk composites combined with a straightforward preparation method render them suitable as porous materials and scaffolds with tunable compliance and reusable nanoprobes for real time optical detection in biomedical, environmental, and industrial applications.