A Novel Mechanism of Macrophage Activation by the Natural Yolkin Polypeptide Complex from Egg Yolk

Ageing is accompanied by the inevitable changes in the function of the immune system. It provides increased susceptibility to chronic infections that have a negative impact on the quality of life of older people. Therefore, rejuvenating the aged immunity has become an important research and therapeutic goal. Yolkin, a polypeptide complex isolated from hen egg yolks, possesses immunoregulatory and neuroprotective activity. Considering that macrophages play a key role in pathogen recognition and antigen presentation, we evaluated the impact of yolkin on the phenotype and function of mouse bone marrow-derived macrophages of the BMDM cell line. We determined yolkin bioavailability and the surface co-expression of CD80/CD86 using flow cytometry and IL-6, IL-10, TGF-β and iNOS mRNA expression via real-time PCR. Additionally, the impact of yolkin on the regulation of cytokine expression by MAPK and PI3K/Akt kinases was determined. The stimulation of cells with yolkin induced significant changes in cell morphology and an increase in CD80/CD86 expression. Using pharmaceutical inhibitors of ERK, JNK and PI3K/Akt, we have shown that yolkin is able to activate these kinases to control cytokine mRNA expression. Our results suggest that yolkin is a good regulator of macrophage activity, priming mainly the M1 phenotype. Therefore, it is believed that yolkin possesses significant therapeutic potential and represents a promising possibility for the development of novel immunomodulatory medicine.     

Downregulation of MMP-9 Enhances the Anti-Migratory Effect of Cyclophosphamide in MDA-MB-231 and MCF-7 Breast Cancer Cell Lines

Metastasis is one of the most urgent issues in breast cancer patients. One of the factors necessary in the migration process is the remodeling of the extracellular matrix (ECM). Metalloproteinases (MMPs) can break down the elements of the ECM, which facilitates cell movement. Many highly aggressive tumors are characterized by high levels of MMPs. In the case of breast cancer, the association between MMP-9 and the migration potential and invasiveness of cells has been demonstrated. In addition, reports indicating increased migration of breast cancer cells after the administration of the commonly used cytostatic cyclophosphamide (CP) are particularly disturbing. Hence, our research aimed to assess the effect of CP treatment on MDA-MB-231 and MCF-7 cells and how this response is influenced by the downregulation of the MMP-9 level. The obtained results suggest that CP causes a decrease in the survival of breast cancer cells of various invasiveness, and the downregulation of MMP-9 enhances this effect, mainly by inducing apoptosis. Moreover, in the group of MMP-9 siRNA-transfected CP-treated cells, a more severe reduction in invasion and migration of cells of both lines was observed, as indicated by the migration and invasion transwell assays and Wound healing assay. Hence, we suggest that CP alone may not result in satisfactory therapeutic effects. On the other hand, the use of combination therapy targeting MMP-9, together with the CP, could improve the effectiveness of the treatment. Additionally, we confirmed a relationship between the levels of MMP-9 and cytokeratin 19 (CK19).     

Direct synthesis of fibrous high molecular weight polyethylene using vanadium catalysts supported on an SiO2 ionic liquid system

Polyethylene of fibrous morphology was obtained using Cp₂VCl₂ and VCl₂(salenCl₂) catalysts activated by AlEt₂Cl and AlEtCl₂ and heterogenized on a supported ionic liquid system prepared with SiO₂ and 1‐(3‐triethoxysilyl)propyl‐3‐methylimidazolium chloroaluminate. The fibre length ranges from 15 to 60 µm, depending on the reaction conditions. The polyethylene is characterized by a high molecular weight ((1.1–2.4) × 10⁶ g mol^?1) and a narrow molecular weight distribution (1.4–2.5). It is a linear polymer, properly without branching. The DSC method reveals characteristic changes in melting temperature and crystallinity degree between the first and second scan heating cycles (141 °C and 136 °C, 71% and 46%, respectively). The wide angle X‐ray spectroscopy and Fourier transform infrared spectroscopy methods give crystallinity degree values of 40% and 41%–51%. The influence of type of catalyst precursor, alkylaluminium activator, catalyst/activator molar ratio, reaction time and temperature is discussed. © 2015 Society of Chemical Industry     

Antioxidant Properties and Rutin Content of High Pressure-Treated Raw and Roasted Buckwheat Groats

The effect of high hydrostatic pressure treatment (200 MPa for 2, 4, and 9 min) on the total antioxidant capacity (TAC), reducing capacity (TRC), and rutin content of raw and roasted buckwheat groats was studied by an updated analytical strategy. TAC and TRC were calculated as the overall capacity of hydrophilic and lipophilic antioxidants. Antioxidant capacity was measured against the 2,2′-azinobis-(3-ethylbenzothiazoline-6-sulfonate) radical cation (ABTS^?+) and the 2,2-diphenyl-1-picrylhydrazyl radical (DPPH^?), and by the photo-induced chemiluminescence assay against the superoxide anion radical (O ₂ ^?? ), whereas the reducing capacity was evaluated with the Folin-Ciocalteu reagent (FCR) and directly by the cyclic voltammetry method. Rutin content was determined by HPLC analysis. The TAC of pressure-treated raw and roasted groats was 16–20% and 12.5–17% lower in comparison with untreated groats, respectively. Hydrophilic antioxidants were the main contributor to the TAC of pressure-treated raw and roasted groats. High pressure treatment of raw groats lowered the FCR reducing capacity of hydrophilic antioxidants by an average of 12.6% and 12%, respectively. In contrast, the pressurization of roasted groats increased radical scavenging capacity by 18% on average due to the presence of hydrophilic compounds in roasted groats. The variations in the antioxidant properties of pressure-treated raw groats were accompanied by an average 47% drop in rutin content, whereas irregular changes were noted in HP-treated and roasted material.     

Murine Fibroblasts and Primary Hepatocytes as Tools When Studying the Efficacy of Potential Therapies for Mucopolysaccharidosis Type I

Mucopolysaccharidosis type I (MPS I) is a metabolic genetic disease caused by the deficiency of a lysosomal enzyme involved in glycosaminoglycans (GAGs) degradation. MPS I cells have a constant level of GAG synthesis, but disturbed degradation means that GAGs accumulate progressively, impairing cell metabolism. GAG metabolism can be modulated by flavonoids, and these are being studied as therapeutics for MPS. We have optimised the protocol for obtaining fibroblasts and hepatocytes from the MPS I murine model and characterised the cells for their suitability as an in vitro model for testing compounds with therapeutic potential. Methods: Murine primary hepatocytes and fibroblasts were used as a cellular model to study the effect of genistein, biochanin A, and kaempferol on the modulation of the GAG synthesis process. Flavonoids were used individually as well as in two-component mixtures. There were no statistically significant differences in GAG synthesis levels from cell types obtained from either wild-type or MPS I mice. We also showed that MPS I fibroblasts and hepatocytes store GAGs, which makes them useful in vitro models for testing the effectiveness of substrate reduction therapies. Furthermore, tested flavonoids had a different impact on GAG synthesis depending on cell type and whether they were used alone or in a mixture. The tested flavonoids reduce GAG synthesis more effectively in fibroblasts than in hepatocytes, regardless of whether they are used individually or in a mixture. Flavonoids modulate the level of GAG synthesis differently depending on cell types, therefore in vitro experiments performed to assess the effectiveness of potential therapies for metabolic diseases should be carried out using more than one cell model, and only such an approach will allow for full answering scientific questions.     

Investigation of droplet breakup in liquid-liquid dispersions by CFD-PBM simulations: The influence of the surfactant type

The accurate prediction of the droplet size distribution (DSD) in liquid-liquid turbulent dispersions is of fundamental importance in many industrial applications and it requires suitable kernels in the population balance model.When a surfactant is included in liquid-liquid dispersions,the droplet breakup behavior will change as an effect of the reduction of the interracial tension.Moreover,also the dynamic interfacial tension may be different with respect to the static,due to the fact that the surfactant may be easily desorbed from the droplet surface,generating additional disruptive stresses.In this work,the performance of five breakup kernels from the literature is assessed,to investigate their ability to predict the time evolution of the DSD and of the mean Sauter diameter,when different surfactants are employed.Simulations are performed with the Quadrature Method of Moments for the solution of the population balance model coupled with the two-fluid model implemented in the compressibleTwoPhaseEulerFoam solver of the open-source computational fluid dynamics (CFD) code OpenFOAM v.2.2.x.The time evolution of the mean Sauter diameter predicted by these kernels is validated against experimental data for six test cases referring to a stirred tank with different types of surfactants (Tween 20 and PVA 88％) at different concentrations operating under different stirrer rates.Our results show that for the dispersion containing Tween 20 additional stress is generated,the multifractal breakup kernel properly predicts the DSD evolution,whereas two other kernels predict too fast breakup of droplets covered by adsorbed PVA.Kernels derived originally for bubbles completely fail.     

The Composition of the Cuticular and Internal Free Fatty Acids and Alcohols from <Emphasis Type="Italic">Lucilia sericata</Emphasis> Males and Females

GC, GC-MS, and HPLC-LLSD analyses were used to identify and quantify cuticular and internal lipids in males and females of the blow-fly (Lucilia sericata). Sixteen free fatty acids, seven alcohols and cholesterol were identified and quantitatively determined in the cuticular lipids of L. sericata. Cuticular fatty acids ranged from C(6) to C(20) and included unsaturated entities such as 16:1n-9, 18:1n-9, 20:4n-3 and 20:5n-3. Cuticular alcohols (only saturated and even-numbered) ranged from C(12) to C(20) in males and C(10) to C(22) in females. Only one sterol was found in the cuticular lipids of both males and females. 23 free fatty acids, five alcohols and cholesterol were identified in the internal lipids. Internal fatty acids were present in large amounts-7.4 mg/g (female) and 10.1 mg/g (male). Only traces of internal alcohols (from C(14) to C(26) in males, from C(14) to C(22) in females) were found in L. sericata. Large amounts of internal cholesterol were identified in L. sericata males and females (0.49 and 0.97 mg/g of the insect body, respectively).