Phospholipid Cyclosporine Prodrugs Targeted at Inflammatory Bowel Disease (IBD) Treatment: Design,Synthesis, and in Vitro Validation

Novel phospholipid (PL)-cyclosporine conjugates were prepared and studied as potential prodrugs for inflammatory bowel disease (IBD). Our approach relies on phospholipase A₂ (PLA₂), which is overexpressed in the inflamed intestinal tissues, as the prodrug activator to potentially release cyclosporine at the site of inflammation. PL-cyclosporine prodrug conjugates with methylene linkers of various lengths between the sn-2 position of the PL and cyclosporine were synthesized and evaluated for in vitro activation. Surprisingly, despite previous work indicating that conjugates with six methylene linkers between the lipid and drug would suffer rapid enzymatic hydrolysis, with cyclosporine this was not observed. However, compounds with longer linkers (n=10, 12 methylene units) display complete release of the drug by PLA₂-catalyzed hydrolysis, thus demonstrating the importance and profound impact of structural fine-tuning. This study represents a proof-of-concept for our hypothesis and a first step towards a truly targeted IBD treatment with cyclosporine that could be administered throughout the GI tract.     

Inulin-based emulsion-filled gel as a fat replacer in prebiotic- and PUFA-enriched dry fermented sausages

In order to produce fermented sausages with prebiotic fibre and improved fatty acid composition, 16% of pork back fat was replaced with inulin gelled suspension (I) and inulin linseed oil gelled emulsion (IO). Physico-chemical analysis, fatty acid profiles, lipid oxidation, microbiological, textural, colour and sensory analysis were carried out. The fat content was lower in I (31.38%) and IO (35.36%) modified sausages compared to control (44.37%) (P < 0.05). IO sausages had lower SFA and MUFA and higher PUFA content with an improved n-6/n-3 ratio (2.23) (P < 0.05) and α-linolenic acid increment (5.74 g per 100 g). Reformulation led to decrease in springiness, chewiness and hardness and increase in adhesiveness of the sausages. Modified sausages had lower L* and higher a* values, while b* values of I sausages did not differ compared to control sausages. Modified sausages were acceptable regarding all sensory attributes. Lipid oxidation parameters showed higher susceptibility to oxidation and lipolysis in IO sausages.     

Development of protease nanobiocatalysts and their application in hydrolysis of sunflower meal protein isolate

In this study, the suitability of fumed silica nanoparticles (FNS) and its derivatives (amino-modified FNS (AFNS), cyanuric chloride-activated AFNS (CCAFNS) and epoxy-modified FNS (GFNS)), for covalent immobilisation of two commercial protease preparations Alcalase^® and Flavourzyme^® was investigated. The highest hydrolytic activities of immobilised preparations were 25 IU g⁻¹ support (Alcalase-GFNS) and 2.95 IU g⁻¹ support (Flavourzyme-CCAFNS). Furthermore, the immobilised preparations showed 43% and 20% of initial specific activities of commercial protease preparations, respectively. Flavourzyme-CCAFNS also exhibited the highest exopeptidase activity of 22.83 L-pNAU g⁻¹ support. Finally, these two nanobiocatalysts were successfully applied for hydrolysis of sunflower meal protein isolate (SMPI), providing two times higher hydrolysis yields in comparison to free enzymes, justifying the applied immobilisation process. Namely, the highest hydrolysis yield (30%) was gained by the sequential hydrolysis with Alcalase-GFNS and Flavourzyme-CCAFNS, which resulted in the formation of small hydrophobic and hydrophilic peptides, ≤5 kDa, confirmed by HPLC analysis and electrophoretic separation.     

Macrophage Immunomodulation Through New Polymers that Recapitulate Functional Effects of Itaconate as a Power House of Innate Immunity

Itaconate (ITA) is an emerging powerhouse of innate immunity with therapeutic potential that is limited in its ability to be administered in a soluble form. A library of polyester materials that incorporate ITA into polymer backbones resulting in materials with inherent immunoregulatory behavior is developed. Harnessing hydrolytic degradation release from polyester backbones, ITA polymers result in the mechanism specific immunoregulatory properties on macrophage polarization in vitro. In a functional assay, the polymer-released ITA inhibits bacterial growth on acetate. Translation to an in vivo model of biomaterial associated inflammation, intraperitoneal injection of ITA polymers demonstrate a rapid resolution of inflammation in comparison to a control polymer silicone, demonstrating the value of sustained biomimetic presentation of ITA.     

Whey valorization using transgalactosylation activity of immobilized β-galactosidase

Whey represents a significant dairy industry by-product that has recently received due attention based on the rich nutrient composition and significant transformation potential. Hereby, we investigated a possibility of whey lactose bioconversion into prebiotic compounds, galacto-oligosaccharides (GOS) using β-galactosidase transgalactosylation activity. The results showed that whey could be successfully used for GOS synthesis, since the highest GOS concentration (around 62 g L⁻¹) was obtained batchwise using 40% (w/w) sweet whey powder solution under optimum conditions (50°C, pH 4.5). Nevertheless, an efficient immobilized preparation using methacrylic Lifetech ECR8409 immobilization carrier was developed, enabling additional process improvement and ensuring at least 4 reaction cycles with unchanged yields and 2.5- fold enhanced productivity in comparison to the soluble enzyme. Therefore, this study provides a valuable contribution to the efficient and economical valorization of whey, which can be further on utilized as functional food and feed constituent.     

Thermally Induced Microstructural Transformations of Fe72Si15B8V4Cu1 Alloy

Thermal stability, mechanism, and kinetics of thermally induced microstructural transformations and their effects on magnetic permeability of Fe₇₂Si₁₅B₈V₄Cu₁ alloy with combined amorphous/nanocrystalline structure were studied. DTA curves revealed two separated thermally activated exothermic events in the temperature ranges from 740 K to 820 K (467 °C to 547 °C) and 870 K to 930 K (597 °C to 657 °C). Crystalline phases present in the as-prepared and thermally treated alloy samples were identified, and their microstructural parameters were determined using XRD, while, to gain further insight into the mechanism of microstructural transformations, AFM and SEM–EDS analyses were performed. Deconvolution of the complex DTA peak into individual steps was conducted, and, in correlation with the results of microstructural analysis, kinetic triplets corresponding to individual transformation steps were determined, allowing for the estimation of the lifetimes of the alloy at different temperatures. Magnetic permeability measurements showed that, in spite of the influence of microstructural transformations on magnetic properties of the alloy, the favorable magnetic properties are retained over relatively a wide temperature range.