Sphk1 and Sphk2 Differentially Regulate Erythropoietin Synthesis in Mouse Renal Interstitial Fibroblast-like Cells

Erythropoietin (Epo) is a crucial hormone regulating red blood cell number and consequently the hematocrit. Epo is mainly produced in the kidney by interstitial fibroblast-like cells. Previously, we have shown that in cultures of the immortalized mouse renal fibroblast-like cell line FAIK F3-5, sphingosine 1-phosphate (S1P), by activating S1P₁ and S1P₃ receptors, can stabilize hypoxia-inducible factor (HIF)-2α and upregulate Epo mRNA and protein synthesis. In this study, we have addressed the role of intracellular iS1P derived from sphingosine kinases (Sphk) 1 and 2 on Epo synthesis in F3-5 cells and in mouse primary cultures of renal fibroblasts. We show that stable knockdown of Sphk2 in F3-5 cells increases HIF-2α protein and Epo mRNA and protein levels, while Sphk1 knockdown leads to a reduction of hypoxia-stimulated HIF-2α and Epo protein. A similar effect was obtained using primary cultures of renal fibroblasts isolated from wildtype mice, Sphk1−/−, or Sphk2−/− mice. Furthermore, selective Sphk2 inhibitors mimicked the effect of genetic Sphk2 depletion and also upregulated HIF-2α and Epo protein levels. The combined blockade of Sphk1 and Sphk2, using Sphk2−/− renal fibroblasts treated with the Sphk1 inhibitor PF543, resulted in reduced HIF-2α and Epo compared to the untreated Sphk2−/− cells. Exogenous sphingosine (Sph) enhanced HIF-2α and Epo, and this was abolished by the combined treatment with the selective S1P₁ and S1P₃ antagonists NIBR-0213 and TY52156, suggesting that Sph was taken up by cells and converted to iS1P and exported to then act in an autocrine manner through S1P₁ and S1P₃. The upregulation of HIF-2α and Epo synthesis by Sphk2 knockdown was confirmed in the human hepatoma cell line Hep3B, which is well-established to upregulate Epo production under hypoxia. In summary, these data show that sphingolipids have diverse effects on Epo synthesis. While accumulation of intracellular Sph reduces Epo synthesis, iS1P will be exported to act through S1P₁₊₃ to enhance Epo synthesis. Furthermore, these data suggest that selective inhibition of Sphk2 is an attractive new option to enhance Epo synthesis and thereby to reduce anemia development in chronic kidney disease.     

Effects and modelling of ultrasonic waste‐activated sludge disintegration

Sonication is a well‐known sludge pretreatment technique with the advantages of simple operation and high efficiency. However, it is an energy‐intensive process. Hence, it is very important to predetermine its sludge disintegration efficiency at varying pretreatment conditions in order to minimize the ultrasonic energy consumption. In this study, it was found that the ultrasonic sludge disintegration occurred in two stages: rapid and subsequent slow disintegration stages. For this reason, it was aimed to develop a simple and accurate mathematical model to describe the two‐stage sludge disintegration as a function of pretreatment conditions. Sludge concentration and ultrasonic density along with sonication period were involved in this model as independent variables. It was determined that the mathematical model can predict accurately the degree of sludge disintegration. Thus, the proposed model was seen to be very useful for evaluating the disintegration efficiency and/or for process design using the operating parameters under different conditions.     

Reflective light modulator based on epsilon-GaSe crystal

We report the results of investigating a low-voltage, polarization-insensitive, reflective-type modulator based on an epsilon-GaSe crystal and operated at the 1.960-eV line of a He-Ne laser. We demonstrate that the modulation in an Al-epsilon-GaSe-Cu device results mainly from the Franz-Keldysh effect. Relatively high speed and low operating voltage could make these modulators with Schottky-barrier contacts attractive devices in the red range of the spectrum.     

Determination of compounding and external load characteristics of saturated cylindrical-rotor synchronous generators

The paper describes a method for predicting the load (compounding or excitation and external) characteristics of saturated cylindrical-rotor synchronous machines. The method is based upon the phasor diagram representation of a synchronous machine and utilizes a simple formula for approximating the magnetisation curve. This yields a closed form of expression for the excitation characteristics, while solution of the equations of the external load characteristics is carried out, easily, by using a simple iteration process. The validity of the approach is verified by tests.     

Mechanical properties of bovine hydroxyapatite (BHA) composites doped with SiO<Subscript>2</Subscript>, MgO,Al<Subscript>2</Subscript>O<Subscript>3</Subscript>, and ZrO<Subscript>2</Subscript>

Biologically derived hydroxyapatite from calcinated (at 850 °C) bovine bones (BHA) was doped with 5 wt% and 10 wt% of SiO₂, MgO, Al₂O₃ and ZrO₂ (stabilized with 8% Y₂O₃). The aim was to improve the sintering ability and the mechanical properties (compression strength and hardness) of the resultant BHA-composites. Cylindrical samples were sintered at several temperatures between 1,000 and 1,300 °C for 4 h in air. The experimental results showed that sintering generally occurs at 1,200 °C. The BHA–MgO composites showed the best sintering performance. In the BHA–SiO₂ composites, extended formation of glassy phase occurred at 1,300 °C, resulting in structural degradation of the resultant samples. No sound reinforcement was achieved in the case of doping with Al₂O₃ and zirconia probably due to the big gap between the optimum sintering temperatures of BHA and these two oxides.     

Synergistic effects of sono-alkaline pretreatment on anaerobic biodegradability of waste activated sludge

The individual and combined effects of alkaline and ultrasonic pretreatment on both physical and chemical properties and anaerobic biodegradability of waste activated sludge (WAS) were investigated comprehensively in this study. The experimental results showed that both disintegration and anaerobic biodegradability of WAS were significantly improved by the combination of alkaline and ultrasonic (sono-alkaline) pretreatment. Besides, it was determined that the hydraulic retention time in anaerobic digester can be shortened by half using this combined pretreatment. However, it was also determined that sono-alkaline pretreatment was not feasible economically due to its high energy requirement.     

Preparation of chitosan‐coated magnetite nanoparticles and application for immobilization of laccase

In this study, immobilization of laccase (L) enzyme on magnetite (Fe₃O₄) nanoparticles was achieved, so that the immobilized enzyme could be used repeatedly. For this purpose, Fe₃O₄ nanoparticles were coated and functionalized with chitosan (CS) and laccase from Trametes versicolor was immobilized onto chitosan‐coated magnetic nanoparticles (Fe₃O₄‐CS) by adsorption or covalent binding after activating the hydroxyl groups of chitosan with carbodiimide (EDAC) or cyanuric chloride (CC). For chitosan‐coated magnetic nanoparticles, the thickness of CS layer was estimated as 1.0–4.8 nm by TEM, isoelectric point was detected as 6.86 by zeta (ζ)‐potential measurements, and the saturation magnetization was determined as 25.2 emu g^?1 by VSM, indicating that these nanoparticles were almost superparamagnetic. For free laccase and immobilized laccase systems, the optimum pH, temperature, and kinetic parameters were investigated; and the change of the activity against repeated use of the immobilized systems were examined. The results indicated that all immobilized systems retained more than 71% of their initial activity at the end of 30 batch uses. © 2011 Wiley Periodicals, Inc. J Appl Polym Sci, 2011