Investigating the cryoprotective efficacy of fructans in mammalian cell systems via a structure-functional perspective.

Fructans have long been known with their role in protecting organisms against various stress factors due to their ability to induce controlled dehydration and support membrane stability. Considering the vital importance of such features in cryo-technologies, this study aimed to explore the cryoprotective efficacy of fructans in mammalian cell systems where structurally different fructan polymers were examined on in vitro cell models derived from organs such as the liver, frequently used in transplantation, osteoblast, and cord cells, commonly employed in cell banking, as well as human seminal fluids that are of vital importance in assisted reproductive technology. To gain insights into the fructan/membrane interplay, structural differences were linked to rheological properties as well as to lipid membrane interactions where both fluorescein leakage from unilamellar liposomes and membrane integrity of osteoblast cells were monitored. High survival rates obtained with human endothelial, osteoblast and liver cells for up to two months clearly showed that fructans could be considered as effective non-permeating cryoprotectants, especially for extended periods of cryopreservation. In trials with human seminal fluid, short chained levan in combination with human serum albumin and glycerol proved very effective in preserving semen samples across multiple patients without any morphological abnormalities.

A comparative study on daily evapotranspiration estimation by using various artificial intelligence techniques and traditional regression calculations.

Evapotranspiration is an important parameter to be considered in hydrology. In the design of water structures, accurate estimation of the amount of evapotranspiration allows for safer designs. Thus, maximum efficiency can be obtained from the structure. In order to accurately estimate evapotranspiration, the parameters affecting evapotranspiration should be well known. There are many factors that affect evapotranspiration. Some of these can be listed as temperature, humidity in the atmosphere, wind speed, pressure and water depth. In this study, models were created for the estimation of the daily evapotranspiration amount by using the simple membership functions and fuzzy rules generation technique (fuzzy-SMRGT), multivariate regression (MR), artificial neural networks (ANNs), adaptive neuro-fuzzy inference system (ANFIS) and support vector regression (SMOReg) methods. Model results were compared with each other and traditional regression calculations. The ET amount was calculated empirically using the Penman-Monteith (PM) method which was taken as a reference equation. In the created models, daily air temperature (T), wind speed (WS), solar radiation (SR), relative humidity (H) and evapotranspiration (ET) data were obtained from the station near Lake Lewisville (Texas, USA). The coefficient of determination (R2), root mean square error (RMSE) and average percentage error (APE) were used to compare the model results. According to the performance criteria, the best model was obtained by Q-MR (quadratic-MR), ANFIS and ANN methods. The R2, RMSE, APE values of the best models were 0,991, 0,213, 18,881% for Q-MR; 0,996; 0,103; 4,340% for ANFIS and 0,998; 0,075; 3,361% for ANN, respectively. The Q-MR, ANFIS and ANN models had slightly better performance than the MLR, P-MR and SMOReg models.

Exploring the potential of Halomonas levan and its derivatives as active ingredients in cosmeceutical and skin regenerating formulations.

Demand on natural products that contain biological ingredients mimicking growth factors and cytokines made natural polysaccharides popular in pharmaceutical and cosmetic industries. Levan is the ß-(2-6) linked, nontoxic, biocompatible, water-soluble, film former fructan polymer that has diverse applications in pharmacy and cosmeceutical industries with its moisturizing, whitening, anti-irritant, anti-aging and slimming activities. Driven by the limited reports on few structurally similar levan polymers, this study presents the first systematic investigation on the effects of structurally different extremophilic Halomonas levan polysaccharides on human skin epidermis cells. In-vitro experiments with microbially produced linear Halomonas levan (HL), its hydrolyzed, (hHL) and sulfonated (ShHL) derivatives as well as enzymatically produced branched levan (EL) revealed increased keratinocyte and fibroblast proliferation (113-118 %), improved skin barrier function through induced expressions of involucrin (2.0 and 6.43 fold changes for HL and EL) and filaggrin (1.74 and 3.89 fold changes for hHL and ShHL) genes and increased type I collagen (2.63 for ShHL) and hyaluronan synthase 3 (1.41 for HL) gene expressions together with fast wound healing ability within 24 h (100 %, HL) on 2D wound models clearly showed that HL and its derivatives have high potential to be used as natural active ingredients in cosmeceutical and skin regenerating formulations.

Sulfated levan from Halomonas smyrnensis as a bioactive, heparin-mimetic glycan for cardiac tissue engineering applications.

Chemical derivatives of levan from Halomonas smyrnensis AAD6(T) with low, medium and high levels of sulfation were synthesized and characterized by FTIR and 2D-NMR. Sulfated levan samples were found to exhibit anticoagulation activity via the intrinsic pathway like heparin in a dose-dependent manner. Exceptionally high heparin equivalent activity of levan sulfate was shown to proceed via thrombin inhibition where decreased Factor Xa activity with increasing concentration was observed in antithrombin tests and above a certain concentration, levan sulfate showed a better inhibitor activity than heparin. In vitro experimental results were then verified in silico by docking studies using equilibrium structures obtained by molecular dynamic simulations and results suggested a sulfation dependent binding mechanism. With its high biocompatibility and heparin mimetic activity, levan sulfate can be considered as a suitable functional biomaterial to design biologically active, functionalized, thin films and engineered smart scaffolds for cardiac tissue engineering applications.

Combinatorial MAPLE gradient thin film assemblies signalling to human osteoblasts.

There is increased interest in smart bioactive materials to control tissue regeneration for the engineering of cell instructive scaffolds. We introduced combinatorial matrix-assisted pulsed laser evaporation (C-MAPLE) as a new method for the fabrication of organic thin films with a compositional gradient. Synchronized C-MAPLE of levan and oxidized levan was employed to assemble a two-compound biopolymer film structure. The gradient of the film composition was validated by fluorescence microscopy. In this study, we investigated the cell response induced by the compositional gradient using imaging of early osteoblast attachment and analysis of signalling phosphoprotein expression. Cells attached along the gradient in direct proportion to oxidized levan concentration. During this process distinct areas of the binary gradient have been shown to modulate the osteoblasts' extracellular signal-regulated kinase signalling with different propensity. The proposed fabrication method results in the preparation of a new bioactive material, which could control the cell signalling response. This approach can be extended to screen new bioactive interfaces for tissue regeneration.