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The bioactivity of the versatile biodegradable biopolymer poly(lactic acid) (PLA) can be obtained by combining it with natural or synthetic compounds. This paper deals with the preparation of bioactive formulations involving the melt processing of PLA loaded with a medicinal plant (sage) and an edible oil (coconut oil), together with an organomodifed montmorillonite nanoclay, and an assessment of the resulting structural, surface, morphological, mechanical, and biological properties of the biocomposites. By modulating the components, the prepared biocomposites show flexibility, both antioxidant and antimicrobial activity, as well as a high degree of cytocompatibility, being capable to induce the cell adherence and proliferation on their surface. Overall, the obtained results suggest that the developed PLA-based biocomposites could potentially be used as bioactive materials in medical applications.

The aim of this study was to obtain biocomposites consisting of poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV), bacterial cellulose (BC) and α-tocopherol by a melt processing technique for potential use in biomedical applications. The melt processing and roughness of biocomposites were evaluated and compared to sample without BC. The degradation rate of PHBV/BC biocomposites was measured in phosphate buffer saline (PBS) by determining the mass variation and evidencing of thermal and structural changes by differential scanning calorimetry (DSC) and attenuated total reflectance-Fourier transformed infrared spectrometry (ATR-FTIR). The cell viability, cell morphology, cell cycle distribution and total collagen content were investigated on murine NCTC fibroblasts. Overall, the adding of BC to polyester matrix led to an adequate melt processing of biocomposites and increased surface roughness and cytocompatibility, allowing the cells to secrete the extracellular matrix (collagen) and stimulate cell proliferation. Results showed that the PHBV/BC biocomposites were favorable for long-term degradation and could be used for the design of medical devices with controlled degradability.

The aim of this study was to obtain biocomposites consisting of poly(3-hydroxybutyrate

Bioactive collagen–chitosan–lemongrass (COL–CS–LG) membranes were prepared by casting method and analyzed for potential biomedical applications. For COL–CS–LG membranes, LG essential oil release, antioxidant properties, in vitro cytotoxicity and antimicrobial assessments were conducted, as well as free radical determination after gamma irradiation by chemiluminescence, and structural characteristics analysis through Attenuated Total Reflection–Fourier Transform Infrared Spectroscopy (ATR–FTIR) and Differential Scanning Calorimetry (DSC). The evaluation of non-isothermal chemiluminescence after gamma radiation exposure to COL–CS–LG membranes revealed a slowing down of the oxidation process at temperatures exceeding 200 °C, in correlation with antioxidant activity. Antimicrobial properties and minimum inhibitory concentrations were found to be in correlation with cytotoxicity limits, offering the optimum composition for designing new biomaterials.

Hashimoto’s thyroiditis (HT) and Graves’ disease (GD) are prevalent autoimmune disorders, representing opposite ends of the clinical spectrum of autoimmune thyroid diseases (AITD). The pathogenesis involves a complex interplay between environment and genes. Specific susceptibility genes have been discovered that predispose to AITD, including thyroid-specific and immune-regulatory genes. Growing evidence has revealed that genetic and epigenetic variants can alter autoantigen presentation during the development of immune tolerance, can enhance self-peptide binding to MHC (major histocompatibility complex), and can amplify stimulation of T- and B-cells. These gene-driven mechanistic discoveries lay the groundwork for novel treatment targets. This review summarizes recent advances in our understanding of key AITD susceptibility genes (Tg, TSHR, HLA-DR3, and CD40) and their translational therapeutic potential.

Programmed death-ligand 1 (PD-L1) is a strong immunosuppressor that inhibits T-cell activation by binding to its receptor (PD-1) on T-cells. Blocking the PD-L1/PD-1 interaction by monoclonal antibodies (mAbs) is an effective therapy for several cancers via activation of T-cells. Destructive thyroiditis and hypothyroidism often develop during anti-PD-L1 (αPD-L1) therapy but the pathogenic mechanisms are not known. We evaluated in vivo, the role of PD-L1 and its interactions with inflammatory triggers in the development of autoimmune thyroiditis (AIT). NOD.H-2h4 mice were treated with either αPD-L1 or isotype control (CTR) for two weeks and AIT evaluated after six weeks. αPD-L1 treated mice showed significantly higher Abs (p = 6×10e-4) and T-cell responses (p = 0.02) to thyroglobulin (Tg) compared with CTR mice. Serum fT4 levels were lower (p = 0.01) and TSH higher (p = 0.05) in αPD-L1 treated vs. CTR mice. Seven of the eight αPD-L1 treated mice (and none of the CTR mice) developed thyroid lymphocytic infiltration characterized by the presence of the CD4+, CD8+, FoxP3+ T- cells and B220+ B- cells. To gain insight into the pathways regulated by PD-L1 in the thyroid we assessed gene expression in the thyroid of αPD-L1 treated mice using RNA-seq. PD-L1 blockade was associated with upregulation of pathways of lymphocyte activation (p = 1.3×10e-5), lymphocyte proliferation (p = 2×10e-5), chemotaxis (p = 3.8×10e-12), and inflammatory response (p = 7.9×10e-7). Specifically, blocking PD-L1 was associated with upregulation of cytokine/chemokine genes that have a critical role in Th1 differentiation: IFNγ, IL-12, IL-15, CCL1 to CCL5, CD300, CXCL10, CXCL11. Ingenuity Pathway Analysis identified IFNγ and TNF as the top regulators of these activated gene networks. These results indicate that blockade of PD-L1 in thyroid promoted the Th1 mediator, IFNγ, inducing activation and proliferation of thyroidal CD4+ and CD8+ T cells, triggering thyroid tissue damage and AIT. Since PD-L1 mediates immune responses to inflammatory stressors (e.g. viral infections), we evaluated PD-L1 interaction with a known inflammatory trigger of AIT, interferon alpha (IFNα). Treatment with αPD-L1 + IFNα accelerated AIT in NOD.H-2h4 mice that developed high Tg Ab and T-cell responses and thyroidal lymphocytic infiltration only two weeks after treatment. Moreover, thyroiditis was more severe in mice receiving αPD-L1 + IFNα than αPD-L1 alone, manifesting by higher T-cell responses (p = 5×10e-3), and significantly more profound hypothyroidism. These results suggest that PDL-1 has a protective role in the thyroid against inflammatory stressors, and that its blockade exposes the thyroid to inflammatory damage. Therefore, in patients undergoing αPD-L1 therapy, inflammatory conditions (e.g. infections) can be a "second hit" that accelerates the onset of thyroiditis and augments thyroidal inflammation. Presentation: Monday, June 13, 2022 12:00 a.m. - 12:15 a.m.

Bone defects due to trauma or diseases causing bone loss have led to an increased demand for new materials that have applications in therapies for bone pathologies. Mesoporous bioactive glasses (MBGs), based on 70SiO2-(26-x)CaO 4P2O5-xCeO2 (x = 0, 1 and 5 mol % for MBGS, MBGS1Ce and MBGS5Ce) were synthesized by evaporation-induced self-assembly (EISA) method. Ceria was selected due to its antimicrobial and antioxidant properties and positive impact on cells differentiation and mineralization. The pore size values of 2.8 nm confirmed the mesoporosity of the obtained bio-glasses. The biocompatibility tests on mouse fibroblast cells (NCTC clone L929) using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and lactate dehydrogenase (LDH) activity assays revealed no cytotoxicity of Cerium containing MBGs. Formation of a hydroxycarbonate apatite (HCA) layer on MBGs surface, e.g. bioactivity of BMGs, was confirmed by XRD, FTIR, SEM and Raman Spectroscopy while presence of Ce3+/Ce4+ in the glasses before body simulated fluid (SBF) immersion was determined by XPS, UV–Vis measurements. The obtained results show that investigated MBGs are potential materials for applications in bone regeneration.

Context Genetic risk factors play a major role in the pathoetiology of autoimmune thyroid diseases (AITD). So far, only common risk variants have been identified in AITD susceptibility genes. Recently, rare genetic variants have emerged as important contributors to complex diseases, and we hypothesized that rare variants play a key role in the genetic susceptibility to AITD. Objective We aimed to identify new rare variants that are associated with familial AITD. Methods We performed deep sequencing of 3 previously mapped AITD-linked loci (10q, 12q, and 14q) in a dataset of 34 families in which AITD clustered (familial AITD). Results We identified 13 rare variants, located in the inositol polyphosphate multikinase (IPMK) gene, that were associated with AITD (ie, both Graves’ disease [GD] and Hashimoto’s thyroiditis [HT]); 2 rare variants, within the dihydrolipoamide S-succinyltransferase (DLST) and zinc-finger FYVE domain-containing protein (ZFYVE1) genes, that were associated with GD only; and 3 rare variants, within the phosphoglycerate mutase 1 pseudogene 5 (PGAM1P5), LOC105369879, and methionine aminopeptidase 2 (METAP2) genes, that were associated with HT only. Conclusion Our study demonstrates that, in addition to common variants, rare variants also contribute to the genetic susceptibility to AITD. We identified new rare variants in 6 AITD susceptibility genes that predispose to familial AITD. Of these, 3 genes, IPMK, ZFYVE1, and METAP2, are mechanistically involved in immune pathways and have been previously shown to be associated with autoimmunity. These genes predispose to thyroid autoimmunity and may serve as potential therapeutic targets in the future.

This chapter deals with some dimensions of strategic management accounting, including the instruments with which they operate and interconnect in order to make managerial decisions. The main objectives of this chapter are to present the general approaches of the concept of strategic management accounting, the dimensions of the application of strategic management accounting, and the strategic tools used by companies in establishing strategies and making decisions. Based on the specialized literature, the authors present the most significant theoretical-methodological approaches to strategic management accounting, the participation of management accountants in the decision-making process, the tools used by strategic management accounting. All aspects presented by the authors are based on existing literature, university studies, and specialized literature from around the world. The chapter concludes with the conclusions of the authors regarding the dimensions and implications of using the specific tools of strategic management accounting for decision making at managerial level.

Introduction: Nutraceuticals are food or parts of a food that provide medicinal or health benefits, including the prevention and treatment of disease. [...]

BACKGROUND Frail older surgical patients face more than a two-fold increase in postoperative complications, including myocardial infarction, deep vein thrombosis, pulmonary embolism, pneumonia, ileus, and others. Many of these complications occur because of postoperative loss of stamina and poor mobility. Preoperative exercise may better prepare these vulnerable patients for surgery. OBJECTIVE We present the protocol for our ongoing randomized trial to assess the impact of a preoperative walking intervention with remote coaching and pedometer on outcomes of stamina (six-minute walk distance- 6MWD) and mobility (postoperative steps) in older adults with frailty traits. METHODS We plan to conduct a randomized clinical trial. Eligible patients are age 60 or higher, scoring 4 or greater on the Edmonton Frailty Scale assessment, and undergoing a surgical operation that requires a 2 or more night hospital stay. Using block randomization stratified on baseline 6MWD, we assign patients to wear a pedometer. At the end of three baseline days, an athletic trainer (AT) provides a daily step count goal reflecting a 10-20% increase from baseline. Subsequently, the AT calls weekly to further titrate the goal or calls more frequently if the patient is not meeting the prescribed goal. Controls receive general walking advice. Our main outcome is change in 6MWD on postoperative day (POD) 2/3 vs. baseline. We also collect 6MWD approximately 4 weeks after surgery and daily in-hospital steps. RESULTS After an initial pilot phase that ran from June 2019 to December 2019, as of July 2020, we have enrolled 14 patients. We have a total of 6 intervention patients and 8 control patients participating in the trial. Funding has been secured till December 2020. CONCLUSIONS If changes in 6MWD and step counts are significantly higher for the intervention group, we believe this will confirm our hypothesis that the intervention leads to decreased loss of stamina and mobility. Once confirmed, we anticipate expanding to multiple centers to assess the interventional impact on clinical endpoints. CLINICALTRIAL The randomized clinical trial was registered on clinicaltrials.gov under the identifier NCT03892187 on March 27, 2019. URL: https://clinicaltrials.gov/ct2/show/NCT03892187

Background: Thyroglobulin (TG) is a key autoantigen in autoimmune thyroid diseases (AITD). Several single nucleotide polymorphisms (SNPs) in the TG locus were shown to be strongly associated with disease susceptibility in both humans and mice, and autoimmune response to TG is the earliest event in the development of thyroid autoimmunity in mice. The classical model of experimental autoimmune thyroiditis (EAT) is induced by immunizing mice with TG protein together with an adjuvant to break down immune tolerance. The classical EAT model has limited utility in genetic studies of TG since it does not allow testing the effects of TG sequence variants on the development of autoimmune thyroiditis. In this study, we have immunized CBA-J mice, an EAT-susceptible strain, with an adenovirus vector encoding the full-length human TG (hTG) to generate a model of EAT in which the TG sequence can be manipulated to test AITD-associated TG SNPs. Methods: We immunized CBA-J mice with hTG-expressing adenovirus following the well-recognized experimental autoimmune Graves' disease protocol that also uses an adenovirus vector to deliver the immunogen. Results: After hTG adenovirus immunizations, mice developed higher T cell proliferative and cytokine responses to hTG and TG2098 (a major T cell epitope in AITD) and higher titers of TG and thyroperoxidase autoantibodies compared with mice immunized with control LacZ-expressing adenovirus. The mice, however, did not develop thyroidal lymphocytic infiltration and hypothyroidism. Conclusions: Our data describe a novel murine model of autoimmune thyroiditis that does not require the use of adjuvants to break down tolerance and that will allow investigators to test the effects of hTG variants in the pathoetiology of Hashimoto's thyroiditis.

Bionanocomposites including poly(lactic acid) (PLA), collagen, and silver nanoparticles (AgNPs) were prepared as biocompatible and stable films. Thermal properties of the PLA-based bionanocomposites indicated an increase in the crystallinity of PLA plasticized due to a small quantity of AgNPs. The results on the stability study indicate the promising contribution of the AgNPs on the durability of PLA-based bionanocomposites. In vitro biocompatibility conducted on the mouse fibroblast cell line NCTC, clone 929, using the 3-(4,5-dimethylthiazol-2- yl)-2,5-diphenyltetrazolium bromide (MTT) assay showed high values of cell viability (&gt, 80%) after cell cultivation in the presence of bionanocomposite formulations for 48 h, while the percentages of lactate dehydrogenase (LDH) released in the culture medium were reduced (&lt, 15%), indicating no damages of the cell membranes. In addition, cell cycle analysis assessed by flow cytometry indicated that all tested bionanocomposites did not affect cell proliferation and maintained the normal growth rate of cells. The obtained results recommend the potential use of PLA-based bionanocomposites for biomedical coatings.

Abstract The influence of the melt flow on the solidification structure is bilateral. The flow plays an important role in the solidification pattern, via the heat transfer, grain distribution, and segregations. On the other hand, the crystal structure, columnar or equiaxed, impacts the flow, via the thermosolutal convection, the drag force applied by the crystals on the melt flow, etc. As the aim of this research was to further explore the solidification–flow interaction, experiments were conducted in a cast cell (95 * 95 * 30 mm3), in which an ammonium chloride–water solution (between 27 and 31 wt pct NH4Cl) was observed as it solidified. The kinetic energy (KE) of the flow and the average flow velocity were calculated throughout the process. Measurements of the volume extension of the mush in the cell and the velocity of the solid front were also taken during the solidification experiment. During the mainly columnar experiments (8 cm liquid height) the flow KE continuously decreased over time. However, during the later series of experiments at higher liquid height (9.5 cm), the flow KE evolution presented a strong peak shortly after the start of solidification. This increase in the total flow KE correlated with the presence of falling equiaxed crystals. Generally, a clear correlation between the strength of the flow and the occurrence of equiaxed crystals was evident. The analysis of the results strongly suggests a fragmentation origin of equiaxed crystals appearing in the melt. The transition from purely columnar growth to a strongly equiaxed rain (CET) was found to be triggered by (a) the magnitude of the coupling between the flow intensity driven by the equiaxed crystals, and (b) the release and transport of the fragments by the same flow recirculating within the mushy zone. Graphical Abstract Coupling mechanism at the origin of CET: 1-2 strong flow running through the mush transporting out dendrite fragments (white dots); 3-4 equiaxed growth and drag of the downward flow. If the vortex is sufficiently stable, the horizontal configuration can lead to freckle appearing; a) vertical solidification front; b) horizontal solidification front.

It is well known that the growth and motion of equiaxed crystals govern important microstructural features, especially in larger castings such as heavy ingots. To determine the origin of the equiaxed crystals, heterogeneous nucleation, and/or fragmentation of dendrite arms from columnar regions are often discussed. In the present study, we demonstrate that under certain conditions relatively large areas of mushy regions slide downward and form spectacular crystal avalanches. These avalanches crumble into thousands of dendritic fragments, whereby the larger fragments immediately sediment and the smaller proceed to behave as equiaxed crystals. Traces of such crystal avalanches can be seen by conspicuous equiaxed layers in the lower part of the casting. From the arguments in the discussion, it is believed that such a phenomenon may occur in alloys which reveal an upward solutal buoyancy in the interdendritic mush. This would include certain steels and other alloys such as Cu-Al, Pb-Sn, or Ni-Al-alloys. Moreover, the occurrence of crystal avalanches contribute to the formation of V-segregations.

The influence of spore concentration on the ability of a Trichoderma consortium to colonize the Passiflora caerulea phyllosphere was evaluated by determining the effects of foliar treatments with two spore concentrations, in two repeated treatments, on the morphological, physiological, and ultrastructural characteristics, and on the yield and quality of P. caerulea. The studied crop quality features were related to its nutraceutical use: the accumulation of polyphenols and flavonoids, antioxidant activity, and effects on mouse fibroblast L929 cells. The Trichoderma consortium consisted of two strains, T. asperellum T36b and T. harzianum Td50b, and the concentrations used were 106 colony forming units (cfu)/mL and 108 cfu/mL. As a reference treatment, a commercial product that was based on herbs and algal extracts was used. As compared to the negative control, the treatment with the Trichoderma consortium at 108 cfu/mL concentration determines the accumulation of higher level of polyphenols and flavonoids and increased antioxidant activity. This enhancement of P. caerulea quality characteristics after treatment with the higher concentration of Trichoderma consortium was associated with larger leaves, increased number and size of chloroplasts, improved plant physiology characteristics, and an increased yield. The treatment with high concentration of Trichoderma consortium spores promotes phyllosphere colonization and benefits both crop yield and quality.

Concentrated collagen hydrolysate (HC10CC), rabbit collagen glue (RCG), and keratin hydrolysate (KH) were investigated in terms of their extraction from mammalian by-products and processing by electrospinning. The electrospun nanofibers were characterized by scanning electron microscopy coupled with the energy dispersive X-ray spectroscopy (SEM/EDS), attenuated total reflectance Fourier transform infrared spectroscopy (ATR-FTIR), differential scanning calorimetry (DSC), and indentation tests. The cytotoxicity of the electrospun nanofibers was conducted on L929 fibroblast cells using MTT and LDH assays and cell morphology observations. The electrospun RCG and KH nanofibers morphology showed an average size of nanofibers ranging between 44 and 410 nm, while the electrospun HC10CC nanofibers exhibited higher sizes. The ATR-FTIR spectra performed both on extracted proteins and electrospun nanofibers showed that the triple helix structure of collagen is partially preserved. The results were in agreement with the circular dichroism analysis for protein extracts. Furthermore, the viscoelastic properties of electrospun KH nanofibers were superior to those of electrospun RCG nanofibers. Based on both in vitro quantitative and qualitative analysis, the electrospun nanofibers were not cytotoxic, inducing a healthy cellular response. The results of new electrospun protein-based nanofibers may be useful for further research on bioactive properties of these nanofibers for tissue engineering.

This chapter provides a business perspective on the policies and benefits provided by implementing environmental managerial accounting in various areas of activity. The main objectives of this chapter are to present the evolution of the EMA and its policies, business analyses carried out by specialists, including the benefits of implementing the EMA. All of these objectives are transposed into a case study based on the ABC method performed at an energy company. The implementation steps specific to the ABC method are presented by establishing the list of main activities, cost drivers, allocation of indirect costs, and determining unit costs. The theoretical aspects presented the basic national and international studies in the literature. The case study is based on data obtained from the energy company. The chapter ends with the authors' conclusions on the benefits of implementing EMA/ABC. This chapter contributes to the expansion of the theoretical and empirical framework of EMA and the methods used to implement it within various business companies.

The association of natural or synthetic polymers with metal nanoparticles resulted in hybrid [...]

Climate change is driving species' range shifts, which are in turn disrupting species interactions due to species-specific differences in their abilities to migrate in response to climate. We evaluated the consequences of asynchronous range shifts in an alpine plant-pollinator community by transplanting replicated alpine meadow turfs downslope along an elevational gradient thereby introducing them to warmer climates and novel plant and pollinator communities. We asked how these novel plant-pollinator interactions affect plant reproduction. We found that pollinator communities differed substantially across the elevation/temperature gradient, suggesting that these plants will likely interact with different pollinator communities with warming climate. Contrary to the expectation that floral visitation would increase monotonically with warmer temperatures at lower elevations, visitation rate to the transplanted communities peaked under intermediate warming at midelevation sites. In contrast, visitation rate generally increased with temperature for the local, lower elevation plant communities surrounding the experimental alpine turfs. For two of three focal plant species in the transplanted high-elevation community, reproduction declined at warmer sites. For these species, reproduction appears to be dependent on pollinator identity such that reduced reproduction may be attributable to decreased visitation from key pollinator species, such as bumble bees, at warmer sites. Reproduction in the third focal species appears to be primarily driven by overall pollinator visitation rate, regardless of pollinator identity. Taken together, the results suggest climate warming can indirectly affect plant reproduction via changes in plant-pollinator interactions. More broadly, the experiment provides a case study for predicting the outcome of novel species interactions formed under changing climates.

In this study, we aimed to obtain gelatin from the marine snail Rapana venosa using acidic and enzymatic extraction methods and to characterize these natural products for cosmetic and pharmaceutical applications. Marine gelatins presented protein values and hydroxyproline content similar to those of commercial mammalian gelatin, but with higher melting temperatures. Their electrophoretic profile and Fourier transform infrared (FTIR) spectra revealed protein and absorption bands situated in the amide region, specific for gelatin molecule. Scanning electron microscopy (SEM) analysis showed significant differences in the structure of the lyophilized samples, depending on the type of gelatin. In vitro studies performed on human keratinocytes showed no cytotoxic effect of acid-extracted gelatin at all tested concentrations and moderate cytotoxicity of enzymatic extracted gelatin at concentrations higher than 0.5 mg/mL. Also, both marine gelatins favored keratinocyte cell adhesion. No irritant potential was recorded as the level of IL-1α and IL-6 proinflammatory cytokines released by HaCaT cells cultivated in the presence of marine gelatins was significantly reduced. Together, these data suggest that marine snails are an alternative source of gelatins with potential use in pharmaceutical and skincare products.

Type 1 diabetes (T1D) is caused by autoimmune destruction of pancreatic β cells. Mounting evidence supports a central role for β cell alterations in triggering the activation of self-reactive T cells in T1D. However, the early deleterious events that occur in β cells, underpinning islet autoimmunity, are not known. We hypothesized that epigenetic modifications induced in β cells by inflammatory mediators play a key role in initiating the autoimmune response. We analyzed DNA methylation (DNAm) patterns and gene expression in human islets exposed to IFN-α, a cytokine associated with T1D development. We found that IFN-α triggers DNA demethylation and increases expression of genes controlling inflammatory and immune pathways. We then demonstrated that DNA demethylation was caused by upregulation of the exoribonuclease, PNPase old-35 (PNPT1), which caused degradation of miR-26a. This in turn promoted the upregulation of ten-eleven translocation 2 (TET2) enzyme and increased 5-hydroxymethylcytosine levels in human islets and pancreatic β cells. Moreover, we showed that specific IFN-α expression in the β cells of IFNα–INS1(CreERT2) transgenic mice led to development of T1D that was preceded by increased islet DNA hydroxymethylation through a PNPT1/TET2–dependent mechanism. Our results suggest a new mechanism through which IFN-α regulates DNAm in β cells, leading to changes in expression of genes in inflammatory and immune pathways that can initiate islet autoimmunity in T1D.

Severe skin injuries, including burns, represent a real concern for the global health-care system and therefore, there is an increased interest in developing wound dressings, in order to stimulate and enhance skin tissue repair. The aim of this study was to design novel hybrid materials, biomimetic to skin extracellular matrix and enriched with silver nanoparticles (nAg), in order to provide both dermal tissue regeneration and antimicrobial activity. Two material variants (variant A and variant B) consisting of type I collagen (COL), chondroitin sulfate (CS) and k-elastin peptides (EL) enriched with positively-charged nAg, were conditioned as membranes. UV exposure ensured both sterilisation and cross-linking of the materials. Physico-chemical characterization of the hybrid biomaterials showed values of density and swelling degree higher than those of COL membrane, while the process of in vitro degradation followed a similar pattern. Infrared spectroscopy and x-ray diffraction indicated alterations of the characteristic structural features and crystallinity of COL after blending with CS and EL and nAg embedding. Scanning electron microscopy observations revealed different surface morphologies of the hybrid membranes, according to their composition. In vitro studies on L929 fibroblasts and HaCaT keratinocytes showed that both hybrid membranes exhibited good cytocompatibility and promoted higher cell proliferation compared to COL sample, as evaluated by MTT and Live/Dead assays. The presence of actin filaments highlighted by fluorescent labelling confirmed the fibroblast and keratinocyte adhesion onto the surface of hybrid membranes. Most importantly, both materials showed an increased wound healing ability in an in vitro scratch assay model, stimulating cell migration at 24 h post-seeding. In addition, good antimicrobial activity was recorded, especially against Gram-positive bacterial strain. Altogether, our findings recommend COL-CS-EL-nAg hybrid membranes as good candidates for wound healing acceleration and bioengineering of skin tissue.

CD40, a tumor necrosis factor receptor, is a major immune-modulating susceptibility gene for Graves disease (GD) as well as for a variety of other autoimmune diseases. Its broad association with autoimmunity underscores its paramount role in the development of a normal adaptive immune response, primarily in coordinating effective antigen presentation. The molecular pathways by which CD40 activation in the thyroid induces GD are unknown. In this study, we investigated whether NF-κB, a ubiquitious family of transcription factors, mediates the downstream effects of thyroid-specific CD40 activation. Cultured primary human thyrocytes, from patients with and without GD, underwent CD40 stimulation. Once stimulated, cytokines and transcription factors specific for either the canonical nuclear factor κB (NF-κB)1 pathway [interleukin (IL)-6, IL-8, tumor necrosis factor (TNF)-α], which primarily recruits cells for innate immunity, or the noncanonical NF-κB2 pathway [B cell-activating factor of the TNF family, CC chemokine ligand (CCL)21], which directs B cell viability, were analyzed. Significant upregulation in the messenger RNA and protein levels of both canonical and noncanonical pathway cytokines was observed. Western blot analyses of the specific transcription factors for the NF-κB1 and NF-κB2 pathways (p65 and p100/p52, respectively) demonstrated that p65 is constitutively expressed. In contrast, CD40 stimulation robustly increased the expression of the NF-κB2 p52 transcription factor, and the upregulation was significantly more profound in the GD tissue than in the normal thyroid tissue. Our data show that CD40 activity in thyrocytes is prominently mediated via NF-κB and furthermore suggest that the NF-κB1 and NF-κB2 pathways both contribute to the triggering and the progression of GD.

The growing scientific interest in exploitation of vegetable bioactives has raised a number of questions regarding their imminent presence in pharmaceutical formulations. This study intends to demonstrate that a dual combination between vegetable oil (e.g. thistle oil, safflower oil, sea buckthorn oil) and a carrot extract represents an optimal approach to formulate safe carrier systems that manifest cell regeneration effect and promising antioxidant and anti-inflammatory activity. Inclusion of both natural actives into lipid carriers imparted a strong negative charge on the nanocarrier surface (up to -45mV) and displayed average sizes of 70nm to 140nm. The entrapment efficiency of carrot extract into nanostructured carriers ranged between 78.3 and 88.3%. The in vitro release study has demonstrated that the entrapment of the extract represents a viable way for an equilibrated release of carotenoids. Besides the excellent antioxidant properties (e.g. scavenging up to 98% of the free oxygen radicals), the results of cellular integrity (e.g. cell viability of 133%) recommend these nanocarriers based on dual carrot extract-bioactive oil as a promising trend for the treatment of certain disorders in which oxidative stress plays a prominent role. In addition, the lipid nanocarriers based on safflower oil and sea buckthorn oil demonstrated an anti-inflammatory effect on LPS induced THP-1 macrophages, by inhibiting the secretion of two pro-inflammatory cytokines, IL-6 and TNF-α.

In this study, sodium alginate film (Alg) was coated with electrospun collagen glue (Col) extracted from rabbit skin waste, loaded with different commercial antimicrobial agents (chitosan, AG425K and ZnONPs) and investigated in terms of morphological, structural and biological properties. The coated nanostructures were characterized using scanning electron microscopy coupled with the energy-dispersive X-ray (SEM/EDS), Attenuated Total Reflectance Fourier-Transform Infrared spectroscopy (ATR FT-IR), and Atomic Force Microscopy (AFM) tests. The cytotoxicity was investigated on murine L929 fibroblasts using 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide salt (MTT) and lactate dehydrogenase (LDH) assays. Microbiological tests were performed against Staphylococcus aureus ATCC 25923, Escherichia coli ATCC 25922, Pseudomonas aeruginosa ATCC 27853 and Candida albicans ATCC 27853 standard strains. In vitro cell culture tests showed a good cytocompatibility of the coated nanostructured systems, except the sample loaded with ZnONPs, which exhibited a highly cytotoxic effect. Alg-Col-ZnONPs nanostructure inhibited the growth and multiplication of the Staphylococcus aureus ATCC 25923 and Escherichia coli ATCC 25922 bacterial strains. The results of new coated nanostructures may be useful for the development of sustainable biomaterials in a circular economy, with bioactive properties for medical wound dressings.