ALBERT

Description: Similar to phosphorylation, transient conjugation of ubiquitin to target proteins (ubiquitination) mediated by the concerted action of ubiquitin ligases and de-ubiquitinating enzymes (DUBs) can affect substrate function. As obligate intracellular parasites, viruses rely on different cellular pathways for their own replication and the well conserved ubiquitin conjugating/de-conjugating system is not an exception. Viruses not only usurp the host proteins involved in the ubiquitination/de-ubiquitination process, but they also encode their own ubiquitin ligases and DUBs. Here we report that an N-terminal variant of the herpes simplex virus (HSV) type-1 large tegument protein VP1/2 (VP1/2 1–767 ), encompassing an active DUB domain (herpesvirus tegument ubiquitin specific protease, htUSP), and TSG101, a component of the endosomal sorting complex required for transport (ESCRT)-I, functionally interact. In particular, VP1/2 1–767 modulates TSG101 ubiquitination and influences its intracellular distribution. Given the role played by the ESCRT machinery in crucial steps of both cellular pathways and viral life cycle, the identification of TSG101 as a cellular target for the HSV-1 specific de-ubiquitinating enzyme contributes to the clarification of the still under debate function of viral encoded DUBs highly conserved throughout the Herpesviridae family. This article is protected by copyright. All rights reserved

Description: The transition to a pathological phenotype such as Barrett's esophagus occurs via induction of resistance upon repeated contact with gastric refluxate in esophagus. This study examined the molecular changes within normal esophageal epithelial cells (EECs) under short-term acid loading and the role of these changes in defensive resistance against acidic cytotoxicity. After primary cultured EECs were exposed to pH 4-acidified medium (AM4), cell viability was determined by the MTT assay. Reactive oxygen species (ROS) and NAD(P)H oxidase (NOX) activity were measured. Activation of the mitogen-activated protein kinases (MAPKs) MEK/ERK1/2, p38 and JNK; phosphoinositol-3-kinase (PI3K)/Akt, and nuclear factor-kappa B (NF-κB) were detected by Western blot analysis or immunofluorescence staining. AM4 incubation induced intracellular ROS generation accompanied by increase in NOX activity, which was further increased by Na + /H + exchange-1 (NHE1)-dependent inhibition but was prevented by inhibition of NOX or mitochondria complex I. AM4 also induced phosphorylation of MEK/ERK1/2, p38 MAPK, PI3K/Akt, and nuclear translocation of NF-κB, and all these effects, except for p38 MAPK phosphorylation, were abolished by inhibition of ROS. ROS-dependent PI3K/Akt activation, which mediates NF-κB nuclear translocation, was inhibited by protein tyrosine kinase (PTK) inhibitors and NHE1-specific inhibitor. All inhibitors of NHE, ROS, PTK, PI3K or NF-κB further decreased AM4-induced cell viability. Acid loading in the presence of NHE1-dependent protection induced ROS generation by activating NOX and mitochondria complex I, which stimulated PTK/PI3K/Akt/NF-κB-dependent survival in EEC. Our data indicate that normal EEC initially respond to acid loading through intrinsic survival activation. This article is protected by copyright. All rights reserved

Description: The possibility of predicting the droplet size distribution from the particle size distribution was investigated. For that purpose, suspensions of different types of materials were dried in a laboratory-scale spray drier. Drying of suspensions was performed with different sizes of two-fluid nozzles. Droplet size distribution was evaluated from the data obtained for spray drying of bismuth molybdate suspension. The method was validated experimentally with other tested materials. Investigated systems involve processes of drying, crystallization, and coating. The proposed methodology can be applied when nonagglomerated particles, spherical particles, or spherical agglomerates were obtained by spray drying. Spray drying is a unique drying process since formation of solid phase and drying occur simultaneously. Droplet size distribution produced by a two-fluid nozzle during spray drying of bismuth molybdate suspension was evaluated simply from the particle size distribution. The proposed methodology enables prediction of the particle size distribution obtained by spray drying.

Description: The influence of vibration parameters on the segregation phenomenon of a binary mixture in a vibration fluidized bed is investigated. Initially, the mixture composed of spherical balls with different densities but same diameter is in a perfect mixing state in the bed. The motion of particles is simulated based on the discrete element method. The effects of friction coefficient, vibration frequency, amplitudes, and gas velocity are analyzed. The coefficient of segregation to the degree of particle segregation is calculated for different operating conditions. The segregation degree in the vibration fluidized bed is found to be higher than that in the bed without vibration. The curve for the segregation degree exhibits a single peak value which represents the optimal segregation result. The impact of vibration on gas phase- and particle flow is not yet clarified due to the complex motion of particles in vibrated fluidized beds. Discrete particle simulation is used to investigate the segregation behavior of binary mixtures in a fluidized bed. The effect of vibration parameters and suitable operating conditions for segregation, particle distribution, and particle motion are evaluated.

Description: The effects of a turbulence-generating grid on fluid mixing and a passive chemical reaction are experimentally investigated in a liquid shear mixing layer under a nonpremixed condition. The grid is installed at three streamwise locations to find the optimal location to promote the chemical reaction. The results show that the grid generates disturbances at small scales that enhance fluid mixing and the chemical reaction. However, the turbulence intensity and mass diffusion in the mixing layers with the grid decrease rapidly and become even smaller than those in the mixing layer without the grid in a downstream region. Therefore, in the present study, the chemical production is maximized when the grid is installed at where the flow is turning to a developed mixing layer. Installation of a turbulence-generating grid is advantageous to promote fluid mixing and chemical reaction in a free shear flow in which the reactive fluids are introduced under a nonpremixed condition. Special attention is needed regarding the location of the grid to maximize the promotion effect since it could be better to install it in the downstream region rather than the upstream region.

Description: Bubble splitting in 2D gas-solid freely bubbling fluidized beds is experimentally investigated using digital image analysis. The quantitative results can be applied for the development of a new breakage model for bubbly fluidized beds, especially discrete bubble models. The variation of splitting frequency with bubble diameter, new resulting bubble volumes, positions, and also the assumptions of mass and momentum conservation for bubbles after breakage are studied in detail. Small bubbles are found to be more stable than large ones and nearly all mother bubbles split into two almost equally sized daughter bubbles. The momentum of gas bubbles in the vertical direction remains approximately constant after breakage, whereas that of bubbles in the horizontal direction changes with no clear trend. The effect of fluidizing gas velocity in breakage frequency is also examined. The behavior of gas bubbles plays a key role in heat- and mass-transfer operations in fluidized beds. Bubble splitting for Geldart B-type particles under varying conditions was investigated in a pseudo-2D gas-solid fluidized bed by digital image analysis. The quantitative results can facilitate the development of new breakage models for bubbly fluidized beds and are particularly relevant for discrete bubble modeling.

Description: For the application of microreactors in industrial processes, more scaling-up strategies are still required except of the original concept, the numbering-up. A more maneuverable and economic approach, combining both similarity-up and numbering-up, is introduced as an example for an innovative strategy. CFD simulation is also applied to assist analysis and optimization of fluid distribution. Based on this concept, named similarity-up + numbering-up + simulation, a pilot-plant microsieve dispersion minireactor used to proceed the reaction of cyclohexanecarboxylic acid and oleum is designed and tested experimentally. With the contribution of similarity-up, the treating capacity of the core unit is significantly increased. Ten parallel units are integrated to form a correspondingly enhanced capacity of the microreactor which exhibits low pressure drop, stable operating performance, and excellent main product selectivity. For industrial applications of microreactors, more flexible and economic scaling-up strategies are required. A scale-up approach for a microsieve dispersion reactor is proposed, combining similarity-up and numbering-up of the core unit with CFD simulation-based device structure optimization. Low fabrication cost, low pressure drop, and excellent main product selectivity can be achieved by this strategy.

Description: Osteolytic bone diseases are characterized by excessive osteoclast formation and activation. Protein kinase C (PKC)-dependent pathways regulate cell growth, differentiation and apoptosis in many cellular systems, and have been implicated in cancer development and osteoclast formation. A number of PKC inhibitors with anti-cancer properties have been developed, but whether they might also influence osteolysis (a common complication of bone invading cancers) is unclear. We studied the effects of the PKC inhibitor compound, GF109203X on osteoclast formation and activity, processes driven by receptor activator of NFκB ligand (RANKL). We found that GF109203X strongly and dose dependently suppresses osteoclastogenesis and osteoclast activity in RANKL-treated primary mouse bone marrow cells. Consistent with this GF109203X reduced expression of key osteoclastic genes, including cathepsin K, calcitonin receptor, tartrate resistant acid phosphatase (TRAP) and the proton pump subunit V-ATPase-d2 in RANKL-treated primary mouse bone marrow cells. Expression of these proteins is dependent upon RANKL-induced NF-κB and NFAT transcription factor actions; both were reduced in osteoclast progenitor populations by GF109203X treatment, notably NFATc1 levels. Furthermore, we showed that GF109203X inhibits RANKL-induced calcium oscillation. Together, this study shows GF109203X may block osteoclast functions, suggesting that pharmacological blockade of PKC-dependent pathways has therapeutic potential in osteolytic diseases. J. Cell. Physiol. © 2014 Wiley Periodicals, Inc.

Description: The sesquiterpene lactone Parthenolide (PN) exerted a cytotoxic effect on MDA-MB231 cells, a triple-negative breast cancer (TNBC) cell line, but its effectiveness was scarce when employed at low doses. This represents an obstacle for a therapeutic utilization of PN. In order to overcome this difficulty we associated to PN the suberoylanilide hydroxamic acid (SAHA), an histone deacetylase inhibitor. Our results show that SAHA synergistically sensitized MDA-MB231 cells to the cytotoxic effect of PN. It is noteworthy that treatment with PN alone stimulated the survival pathway Akt/mTOR and the consequent nuclear translocation of Nrf2, while treatment with SAHA alone induced autophagic activity. However when the cells were treated with SAHA/PN combination, SAHA suppressed PN effect on Akt/mTOR/Nrf2 pathway, while PN reduced the prosurvival autophagic activity of SAHA. In addition SAHA/PN combination induced GSH depletion, fall in Δψm, release of cytochrome c, activation of caspase 3 and apoptosis. Finally we demonstrated that combined treatment maintained both hyperacetylation of histones H3 and H4 induced by SAHA and down-regulation of DNMT1 expression induced by PN. Inhibition of the DNA-binding activity of NF-kB, which is determined by PN, was also observed after combined treatment. In conclusion, combination of PN to SAHA inhibits the cytoprotective responses induced by the single compounds, but does not alter the mechanisms leading to the cytotoxic effects. Taken together our results suggest that this combination could be a candidate for TNBC therapy. J. Cell. Physiol. © 2014 Wiley Periodicals, Inc.

Description: A new design of conjugated heat transfer in double-pass parallel-plate laminar countercurrent operations of power law fluids under wall isoflux was investigated experimentally and theoretically. The analytical solutions were obtained with a superposition model by introducing an eigenfunction expansion in terms of a power series for the homogeneous part and an asymptotic solution for the inhomogeneous part. The influence of the power law index on the average Nusselt numbers with the various design and operating parameters is also delineated. The theoretical predictions of the experimental results are represented graphically. The heat transfer performance was considerably improved when compared with a single-pass parallel-plate heat exchanger (without inserting a solid separator sheet). Suitable adjustments of the solid separator sheet position can effectively enhance the heat transfer efficiencies for such a recycling double-pass device, as compared with the efficiencies of single- and double-pass devices.

Description: Genomic and trascriptomic profiling has recently contributed details to the characterization of luminal B breast cancer. We explored the contribution of anthropometric, metabolic and molecular determinants to the multifaceted heterogeneity of this breast cancer subtype, with a specific focus on the association between body mass index (BMI), pre-treatment fasting glucose, hormone receptors and expression of human epidermal growth factor receptor 2 (HER2). Extensively annotated specimens were obtained from 154 women with luminal B breast cancer diagnosed at two Italian comprehensive cancer centres. Participants' characteristics were descriptively analyzed overall and by HER2 status (positive vs negative). BMI (〈25 vs ≥25), pre-treatment fasting glucose (〈median value of 94 mg/dl vs ≥94) and percentage of hormone receptors were tested for association with HER2 expression in regression models. In univariate models, BMI, fasting glucose and, at a lesser extent, percentage of estrogen receptors (ER) were significantly and inversely associated with HER2 expression (OR: 0.32, 95%CI: 0.16-0.66; 0.43, 0.23-0.0.82; 0.96, 0.94-0.97, respectively). The multivariate models confirmed the protective role of BMI and ER on HER2 expression, with luminal B HER2 positive patients being significantly less frequent among women within the highest category of BMI and percentage expression of ER compared with their counterparts (OR: 0.22, 95%CI: 0.09-0.53; 0.95, 0.93-0.97). In conclusions, BMI and percentage of ER representation are inversely associated with HER2 expression in luminal B breast cancers. Upon confirmatory findings, this might help identify patient subgroups who may best benefit from the use of interventions targeting insulin resistance in well depicted breast cancer scenarios. This article is protected by copyright. All rights reserved

Description: Histone deacetylases (HDACs) play a crucial role in the regulation of gene expression through remodeling of chromatin structures. However, the molecular mechanisms involved in this event remain unknown. In this study, we sought to examine whether HDAC inhibition-mediated protective effects involved HDAC4 sumoylation, degradation, and the proteasome pathway. Isolated neonatal mouse ventricular myocytes (NMVM) and H9c2 cardiomyoblasts were subjected to 48 hours of hypoxia (H) (1% O 2 ) and 2 hours of reoxygenation (R). Treatment of cardiomyocytes with trichostatin A (TSA) attenuated H/R-elicited injury, as indicated by a reduction of lactate dehydrogenase (LDH) leakage, an increase in cell viability, and decrease in apoptotic positive cardiomyocytes. MG132, a potent proteasome pathway inhibitor, abrogated TSA-induced protective effects, which was associated with the accumulation of ubiquitinated HDAC4. NMVM transduced with adenoviral HDAC4 led to an exaggeration of H/R-induced injury. TSA treatment resulted in a decrease in HDAC4 in cardiomyocytes infected with adenoviral HDAC4, and HDAC4-induced injury was attenuated by TSA. HDAC inhibition resulted in a significant reduction in reactive oxygen species (ROS) in cardiomyoblasts exposed to H/R, which was attenuated by blockade of the proteasome pathway. Cardiomyoblasts carrying wild type and sumoylation mutation (K559R) were established to examine effects of HDAC4 sumoylation and ubiquitination on H/R injury. Disruption of HDAC4 sumoylation brought about HDAC4 accumulation and impairment of HDAC4 ubiquitination in association with enhanced susceptibility of cardiomyoblasts to H/R. Taken together, these results demonstrated that HDAC inhibition stimulates proteasome dependent degradation of HDAC4, which is associated with HDAC4 sumoylation to induce these protective effects. This article is protected by copyright. All rights reserved

Description: In order to study emulsification phenomena, devices generating well-defined flow conditions are essential. Thus, emulsification of drop collectives under laminar shear flow is commonly performed in cylindrical Couette or Searle devices. In these devices, the flow conditions in the shear gap and in the volume underneath the rotor are often different, which can lead to inhomogeneous product properties and may complicate sample taking. Here, a novel cone-cone shear cell is presented to study emulsification processes. The flow inside the device is examined using numerical simulations. The numerical simulations indicate that simple shear flow is realized all over the sample volume in the cone-cone shear cell. The experimental results show that the drop breakup in the cone-cone shear cell is equivalent to the breakup under simple shear realized in the shear gap of a conventional device, i.e., the Searle device. Critical capillary numbers are calculated from the experimental data and show breakup behavior as predicted by single-drop experiments. Thus, the cone-cone shear cell proved to be suitable to study emulsification mechanisms in simple shear flow. Emulsion-based products are widely used in the food, chemical, and pharmaceutical industries. A novel cone-cone shear cell to study emulsification characteristics is described. The flow conditions inside the shear cell are validated via numerical simulations and experimental results.

Description: Hydrocracking of a bitumen-derived asphaltene over NiMo/ γ -Al 2 O 3 was investigated in a microbatch reactor at varying temperatures. The molar kinetics of asphaltene cracking reaction was examined by fitting the experimental data. Below a defined temperature, the molar reaction showed the first-order kinetic feature while at higher temperatures secondary reactions such as coke formation became significant, causing deviation of the reaction behavior from the proposed first-order kinetic model. Selectivity analysis proved that dominant products varied from gases to liquids to gases with increasing temperature, shifting the dominant reaction from C–S bonds cleavage to C–C bonds cleavage. Catalytic hydrocracking could be a proper choice for asphaltene upgrading but the reaction conditions still need to be optimized. Hydrocracking of a bitumen-derived asphaltene over a NiMo/ γ -Al 2 O 3 catalyst in a microbatch reactor at varying temperatures was analyzed. The molar kinetics of asphaltene cracking reaction was evaluated by fitting the experimental data.

Description: Endothelial cells play a major role in the initiation and perpetuation of the inflammatory process in health and disease, including their pivotal role in leukocyte recruitment. The role of pro-inflammatory transcription factors in this process has been well-described, including NF-κB. However, much less is known regarding transcription factors that play an anti-inflammatory role in endothelial cells. Myocyte enhancer factor 2 C (MEF2C) is a transcription factor known to regulate angiogenesis in endothelial cells. Here, we report that MEF2C plays a critical function as an inhibitor of endothelial cell inflammation. Tumor necrosis factor (TNF)-α inhibited MEF2C expression in endothelial cells. Knockdown of MEF2C in endothelial cells resulted in the upregulation of pro-inflammatory molecules and stimulated leukocyte adhesion to endothelial cells. MEF2C knockdown also resulted in NF-κB activation in endothelial cells. Conversely, MEF2C overexpression by adenovirus significantly repressed TNF-α induction of pro-inflammatory molecules, activation of NF-κB, and leukocyte adhesion to endothelial cells. This inhibition of leukocyte adhesion by MEF2C was partially mediated by induction of KLF2. In mice, lipopolysaccharide (LPS)-induced leukocyte adhesion to the retinal vasculature was significantly increased by endothelial cell-specific ablation of MEF2C. Taken together, these results demonstrate that MEF2C is a novel negative regulator of inflammation in endothelial cells and may represent a therapeutic target for vascular inflammation. This article is protected by copyright. All rights reserved

Description: Two different types of metals (Cu and Ni) and the effect of CeO 2 addition to produce a CeO 2 -ZrO 2 co-supporter were investigated through the water-gas shift (WGS) reaction. It was found that the WGS activity could be enhanced with CeO 2 addition. At relatively high temperature, Ni-loaded catalysts exhibited higher CO conversion while Cu-loaded catalysts demonstrated better performance at low temperatures. The stability and yield of the CO 2 and H 2 products of the Cu catalysts were higher than those of the Ni catalysts. These results may be caused by an irreversible adsorption of CO on Ni and the reverse WGS reaction occurring on the Ni catalysts. In situ diffuse-reflection infrared Fourier transform spectroscopy data suggests that the WGS mechanism likely proceeded via formate species. The water-gas shift reaction (WGS), as an alternative way to produce H 2 and convert poisonous CO into CO 2 , was carried out using Cu and Ni loaded onto CeO 2 -ZrO 2 as catalysts. The Ni-loaded catalysts exhibited higher CO conversion at high temperature while the Cu-loaded catalysts performed better at low temperature. The WGS reaction was found to occur via the formation of formate species.

Description: Power ultrasound is applied for cooling crystallization to control and modify the particle size and crystal habit of an active pharmaceutical ingredient, phenacetin. Operating parameters including sonication intensity and duration, solution concentration, and cooling rate are studied and compared. With respect to mean particle size, the effect of sonication intensity is most significant. In addition, the crystal habit of recrystallized phenacetin is modified substantially and shows an elliptic shape. Recrystallized phenacetin also provides an enhanced dissolution rate compared with the original sample. These results prove that ultrasonic crystallization is an efficient tool for controlling the solid-state properties of an active pharmaceutical ingredient. Ultrasonic crystallization is a promising process for controlling the different stages of crystallization. Cooling crystallization applying power ultrasound is adopted for recrystallization of a nonsteroidal anti-inflammatory drug, phenacetin. The mean particle size can be managed by adjusting sonication intensity and duration. Phenacetin crystals with a regular crystal habit and an elliptic shape are obtained.

Description: In a variety of reactions in the chemical industry, eggshell catalysts with a thin active layer are applied; they are often crushed for laboratory testing. The destruction of the shell can be avoided by a special reactor design. The presented advanced TEMKIN reactor is a further development of the reactor system for testing eggshell catalysts on the laboratory scale published by Temkin and Kul'kova in 1969. It is suitable for kinetic studies and for the detailed investigation of deactivation processes, as shown on the example of selective hydrogenation of acetylene. The advanced TEMKIN reactor is well suitable for testing of uncrushed industrial eggshell catalysts because of its defined flow pattern and excellent mass and heat transport properties. Because of its simple and robust design, all technical requirements are fulfilled for a fast, competitive, and accurate optimization of prototypes as well as already established catalysts for industrial applications.

Description: A bench-scale submerged membrane bioreactor (SMBR) was employed to treat vegetable oil plant wastewater with complete sludge retention. Treatment performance and membrane fouling of the SMBR were investigated. The system stably removed high amounts of total organic carbon, oil, and ammonia from vegetable oil wastewater and reduced the chemical oxygen demand, demonstrating the great potential of the SMBR in removing pollutants. The membrane fouling layer was not only governed by deposition of organic substances composed of extracellular polymeric substances like proteins, polysaccharides etc., and oil substances but also by inorganic elements. Organic foulants coupled to inorganic precipitation enhanced the formation of a gel layer and triggered severe membrane fouling in the SMBR. Treatment and disposal of vegetable oil wastewaters (VOWs) represents one of the principal problems for vegetable oil producing countries. A bench-scale submerged membrane bioreactor was applied to treat VOWs with complete sludge retention. Treatment performance and membrane fouling were investigated. The experimental results demonstrated the great potential of this membrane bioreactor in removing pollutants.

Description: Adsorption processes are frequently applied to separate traces of hazardous and toxic substances from gas streams. Hence, knowledge of sorption characteristics of these substances on standard adsorbents is essential. Sorption of hexanal and acetaldehyde from a nitrogen gas stream in trace concentrations on activated carbon and ordered mesoporous carbon-based adsorbents (CMK) is studied. A magnetic suspension balance and an attached gaschromatograph-mass spectrometer were used to analyze the sorption process both gravimetrically and spectrometrically. Both types of adsorbents show a higher capacity for hexanal than acetaldehyde. The activated carbon exhibits considerable differences in regard to desorption of hexanal compared to the mesoporous CMK. Information on sorption characteristics of hazardous substances is essential to design separation processes. Adsorption and desorption of the toxic acetaldehyde and the intensely odorous hexanal on activated carbon and periodic mesoporous carbon were studied. Magnetic suspension balances were used to analyze the sorption processes. Both adsorbents exhibit a higher capacity for hexanal than acetaldehyde.

Description: High-solids biomass slurries exhibit non-Newtonian behavior with a yield stress and require high power input for mixing. The goals were to determine the effect of scale and geometry on power number P 0 , and estimate the power for mixing a pretreated biomass slurry in a 3.8 million L hydrolysis reactor of conventional design. A lab-scale computational fluid dynamics model was validated against experimental data and then scaled up. A pitched-blade turbine and A310 hydrofoil were tested for various geometric arrangements. Flow was transitional; laminar and turbulence models resulted in equivalent P 0 which increased with scale. The ratio of impeller diameter to tank diameter affected P 0 for both impellers, but impeller clearance to tank diameter affected P 0 only for the A310. At least 2 MW is required to operate at this scale. High-solids biomass slurries are characterized by non-Newtonian behavior with a yield stress and high power input demand for mixing. A computational fluid dynamics model was developed to predict power requirements of non-Newtonian lignocellulosic slurry in an industrial-scale hydrolysis reactor with conventional mixing impellers. The lab-scale model was validated against experimental data and then scaled up.

Description: The effect of NaCl added in different quantities on thermodynamic properties, granulometric characteristics, and structure of glycine in a crystallization process was investigated. Solubilities of α - and γ -polymorphs in the presence of varying amounts of NaCl were analyzed. In order to examine the impact of the additive on granulometric properties of glycine, crystal morphology was examined by observing crystals under a scanning electron microscope. Crystal size distribution was determined by sieve analysis. By X-ray diffraction analysis, the critical concentration of NaCl at which the structure of glycine changed, could be defined. The purity of obtained polymorphs was confirmed by Fourier transform infrared spectroscopy. Interactions between additives and crystallizing phase influence crystallization processes. Batch-cooling crystallization of glycine with different amounts of added NaCl is described, causing changes in solubility, metastable zone width, supersaturation, final mass of crystals, granulometric properties, and structure. Process conditions for a conversion of α- into a γ- glycine structure are defined.

Description: Cryogenic air separation as the most important part of an integrated gasification combined cycle is a widely used operation unit for producing large quantities of high-purity oxygen and nitrogen. However, cryogenic distillation requires a large amount of energy due to the work needed to compress the air feed. An improved heat-integrated air separation column (HIASC) is proposed. The requirements of high-purity separation in the industrial cryogenic air separation process are achieved. An optimization model of the heat transfer coefficient ( UA ), a key parameter in column structure design and operation, is presented. The optimized UA value is obtained within the accepted value range reported in the international open literature, which ensures the practicability of the improved HIASC. An improved heat-integrated air separation column is proposed. With the new heat-integrated and thermally coupled structure, the pressure of the high-pressure column and the energy consumption decrease significantly compared with the conventional air separation column. The mathematic model and parameter analysis are presented. An optimization model for the heat transfer coefficient is proposed.

Description: Functional mesoporous Mo–SiO 2 materials were synthesized by a one-pot and facile room-temperature procedure, and characterized by X-ray diffraction, TEM, Raman spectroscopy, FT-IR, diffuse reflectance spectra, and BET analysis. The experimental results demonstrated that the mesoporous materials presented a high dispersion of molybdenum species and excellent catalytic activity for the removal of dibenzothiophene (DBT) without organic solvents as extractants. The catalytic performance on different sulfur-containing compounds was also investigated in detail. After recycling for eight times, the removal of the oxidation desulfurization system could still reach high values. GC-MS analysis detected the oxidation product of DBT. A mechanism was proposed for the absorptive oxidation process of sulfur compounds. The removal of sulfur compounds from petroleum is of utmost importance for stringent fuel specifications and environment pollution. Functional mesoporous Mo–SiO 2 materials were synthesized by a facile procedure and were characterized by a high dispersion of molybdenum species and excellent catalytic activity for the removal of dibenzothiophene under mild conditions without organic solvents as extractants.

Description: The level set method is combined with the concentration transformation method to solve the interphase mass transfer process. However, the artificial diffusion generated in the mass transfer convection term across the interface is inevitable, especially when large shape deformation is encountered at high Reynolds numbers. A semi-Lagrangian advection scheme is introduced to overcome this disadvantage. The methyl isobutyl ketone (MIBK)-acetic acid-water system is adopted to study the unsteady mass transport process accompanied with the Marangoni effect of a single deformable drop ascending in the infinite continuous phase. The predicted overall mass transfer coefficients agree with experimental data very well. The configuration of Marangoni convection is revealed and its effect on the interphase mass transfer process is investigated. The solute-induced Marangoni effect on an ascending drop driven by buoyancy is numerically simulated based on the level set method. The semi-Lagrangian convection scheme is introduced to eliminate the artificial diffusion. Compared with literature data, the present algorithm with the semi-Lagrangian convection scheme significantly suppressed the numerical diffusion and achieved much better predictions.

Description: The surface properties of solvent-based (SB) and water-based (WB) coatings and their impact on fouling during convective heat transfer of CaSO 4 solutions were investigated. Experiments demonstrated that the SB coatings had generally better non-adhesive characteristics, especially at higher values of the electron donor component since the deposits could easily be washed away. For the SB coatings, a longer induction period compared to those of untreated surfaces was observed and a significant reduction of the fouling rate could be achieved. Further analysis of surfaces revealed that SB coatings enhanced the acid-base repulsive force and thus reduced the deposit/solid adhesion energy. For the WB coatings, the Liftshitz-van der Waals attractive force plays a decisive role in the adhesion process due to the higher apolar component of the surface energy. Recent technological advances have given impetus in altering surface properties to mitigate fouling of heat transfer surfaces. The attempted coatings in this study demonstrated that they can extend the induction period of the fouling processes of CaSO 4 deposits by four times. This was but mostly due to their higher electron donor component of the surface energy compared to stainless-steel substrate.

Description: Ultrasonic hot embossing of polymers is an alternative to reduce fabrication costs of microreactors. An ultrasonic welding machine is used to melt a stack of thermoplastic foils and adapting them to a short-time milled aluminum mold showing the inversed design of the desired microfluidic cavities. Two micromixers were fabricated this way providing a low degree of axial dispersion and pressure loss. Stability analysis is successfully performed for a wide temperature range and high pressure. Mixing of colored aqueous solutions and neutralization reactions are implemented to both systems for defined volume flow rates and optically investigated via microscope. Reaction progress is automatically determined with a MATLAB script by reference to the consequential color change of the neutralization reaction with a color indicator. Typical mixing characteristics are identified for both mixers. Ultrasonic hot embossing of chemically resistant polymers as a simple and cost-effective fabrication method of microstructures and an optical analysis tool for tracing the progress of chemical reactions are introduced and evaluated. The simplicity of the fabrication method itself and good similarities of produced polymer micromixers with conventional ones made of other materials are pointed out.

Description: The existing screening and characterization systems for biocatalysts operate in batch mode, which could make catalyst selection and process development inaccurate when continuous operation mode is required in industry. A significant improvement of an innovative screening system based on miniaturized multiple membrane reactors formerly presented by the author is outlined, which enables continuous feeding of substrates and continuous removal of products. Although the presented screening system was originally designed for homogeneous enzymatic reactions, it can be used without further modifications for continuous catalysis with polymer-bound chemical catalysts or for quasi-homogeneous systems like reverse micelles. Currently available screening and characterization systems for biocatalysis are not sufficiently suitable for process description and scale-up of results to pilot- or full-scale reactors often operated in continuous mode. Hydrolysis of N -acetyl- L -methio-nine served as model reaction for an innovative continuous characterization system, implementing a precise dosing system.

Description: The synthesis of n -butyl levulinate, one of the most important biodiesel additives, by catalytic esterification of biomass-derived levulinic acid (LA) with n -butanol over modified H-ZSM-5 (micro/meso-HZ-5) in a closed-batch system is reported for the first time. The optimization of the reaction conditions such as the reactant molar ratio, the catalyst loading, the reaction time and the temperature was performed in view to maximize the yield of n -butyl levulinate. Micro/meso-HZ-5 was found to be the most efficient catalyst, with 98 % yield of n -butyl levulinate and a reusability for six cycles, which is higher than reported in the literature. A possible catalytic mechanism for the esterification reaction is also proposed. A second-order pseudo-homogeneous model with R 2  〉 0.97 confirmed that the esterification reaction is performed in the kinetic regime due to the high activation energy of 23.84 kJ mol −1 . The modified zeolite catalyst H-ZSM-5 (micro/meso-HZ-5) was used as heterogeneous acid catalyst in the esterification of renewable levulinic acid with n -butanol to produce n -butyl levulinate in a closed system. Micro/meso-HZ-5 turned out to be an efficient catalyst with 98 % yield of n -butyl levulinate and high reusability.

Description: Trehalose-6-phosphate phosphatase (TPP) catalyzes the final step in the biosynthesis of the anti-stress sugar trehalose. An 82 kDa TPP enzyme was isolated from Candida utilis with 61% yield and 43-fold purification. The protein sequence, determined by N-terminal sequencing and MALDI-TOF analysis, showed significant homology with known TPP sequences from related organisms. The full length gene sequence of TPP of Candida utilis was identified using rapid amplification of cDNA ends - PCR reaction (RACE-PCR). The gene was cloned and expressed in E. coli BL21. Recombinant TPP enzyme was isolated using affinity chromatography. CD spectroscopy and steady state fluorescence revealed that the structural and conformational aspects were identical in both native and recombinant forms. The biochemical properties of the two forms were also similar. K m was determined to be ∼0.8 mM. Optimum temperature and pH was found to be 30 °C and 8.5 respectively. Activity was dependent on the presence of divalent cations and inhibited by metal chelators. Methylation mediated regulation of TPP enzyme and its effect on the overall survival of the organism under stress were investigated. The results indicated that enhancement of TPP activity by methylation at the Cysteine residues increased resistance of Candida cells against thermal stress. This work involves extensive investigations towards understanding the physico-chemical properties of the first TPP enzyme from any yeast strain. The mechanism by which methylation regulates its activity has also been studied. A correlation between regulation of trehalose synthesis and survivability of the organism under thermal stress was established. J. Cell. Physiol. 9999: XX–XX, 2014. © 2014 Wiley Periodicals, Inc.

Description: The microvascular network of the microcirculation works in tight communication with surrounding tissues to control blood supply and exchange of solutes. In cerebral circulation, microvascular endothelial cells constitute a selective permeability barrier that controls the environment of parenchymal brain tissue, which is known as the blood-brain barrier (BBB). Connexin- and pannexin-formed channels (gap junctions and hemichannels) play a central role in the coordination of endothelial and smooth muscle cell function and connexin-mediated signaling in endothelial cells is essential in the regulation of BBB permeability. Likewise, gap junction communication between astrocyte-end feet also contributes to maintain the BBB integrity, but the participation of hemichannels in this process cannot be discarded. Sympathetic and sensory perivascular nerves are also involved in the control and coordination of vascular function through the release of vasoconstrictor or vasodilator signals and by the regulation of gap junction communication in the vessel wall. Conversely, ATP release through pannexin-1-formed channels mediates the α1-adrenergic signaling. Furthermore, here we show that capsaicin-induced CGRP release from mesenteric perivascular sensory nerves induces pannexin-1-formed channel opening, which in turn leads to reduction of pannexin-1 and endothelial nitric oxide synthase (eNOS) expression along the time. Interestingly, blockade of CGRP receptors with CGRP 8–37 increased eNOS expression by ∼5 fold, suggesting that capsaicin-sensitive sensory nerves are involved in the control of key signaling proteins for vascular function. In this review, we discuss the importance of connexin-based channels in the control of BBB integrity and the functional interaction of vascular connexins and pannexins with the peripheral nervous system.

Description: We previously established a role for cancer-associated fibroblasts (CAFs) in enhancing the self-renewal and differentiation potentials of putative prostate cancer stem cells (CSCs). Our published work focused on androgen-dependent prostate cancer (ADPC) using the conditional Pten deletion mouse model. Employing the same model, we now describe the interaction of CAF and CSC in castration-resistant prostate cancer (CRPC). CAF isolated from ADPC (ADPCAF) and from CRPC (CRPCAF) were compared in terms of their ability to support organoid formation and tumor initiation by CSC from CRPC (CRPCSC) in vitro and in vivo. CRPCSC formed spheroids in vitro and well-differentiated glandular structures under the renal capsules of recipient mice in vivo more effectively in the presence of CRPCAF compared to ADPCAF. Furthermore, whereas CSC with CAF from ADPC formed mostly well-differentiated tumors in our previous study, we now show that CRPCSC, when combined with CRPCAF (but not ADPCAF) can form aggressive, poorly differentiated tumors. The potential of CRPCAF to support organoid/tumor formation by CRPCSC remained greater even when compared to 10-fold more ADPCAF, suggesting that paracrine factors produced specifically by CRPCAF preferentially potentiate the stemness and tumorigenic properties of the corresponding CSC. This apparently unique property of CRPCAF was notable when the CAF and CSC were grafted in either intact or castrated recipient mice. In both environments, CRPCAF induced in the epithelial compartment higher proliferative activity compared to ADPCAF, indicated by a higher Ki67 index. Factors released by CRPCAF to regulate CRPCSC may be targeted to develop novel therapeutic approaches to manage prostate cancer. J. Cell. Physiol. © 2014 Wiley Periodicals, Inc.

Description: Tat transactivating activity regulated by NAD + -dependent histone deacetylase sirtuin1 (SIRT1) connects HIV transcription with the metabolic state of the cell. Nicotinamide phosphoribosyltransferase (Nampt) is a rate-limiting enzyme in the mammalian NAD + biosynthesis. Nampt, SIRT1 and AMPK were involved in inhibiting HIV-1 transactivation through redox-regulated pathway. Tanshinone IIA is a main lipid-soluble monomer derivative from the root of Salvia miltiorrhiza (Danshen) and tanshinone ⅡA possess a variety of biological activities through redox signaling pathway. Here we investigated the effect of tanshinone ⅡA on Tat-induced HIV-1 transactivation and the redox signaling pathway involved in it. As the results were shown, tanshinone ⅡA reversed Tat-induced reactive oxygen species (ROS) production and down-regulation of glutathione (GSH) levels in TZM-bl cells through up-regulation of Nrf2 expression. Tanshinone ⅡA reversed Tat-induced inhibition of SIRT1 activity but not SIRT1 protein expression. Tanshinone ⅡA reversed Tat-induced inhibition of Nampt protein expression and depletion of NAD + levels in TZM-bl cells in a dose-dependent manner. Tanshinone ⅡA-evoked Nampt expression was mediated by AMPK signaling pathway. Tanshinone ⅡA inhibited Tat-induced HIV-1 LTR transactivation dependent on AMPK-Nampt pathway. Collectively, our data provide new insights into understanding of the molecular mechanisms of tanshinone ⅡA inhibited Tat-regulated transcription, suggesting that targeting AMPK/Nampt/SIRT1 pathway could serve as new anti-HIV-1 agents.

Description: Osteoblast differentiation is tightly regulated by several factors including microRNAs (miRNAs). In this paper we report that pre-mir-15b is highly expressed in differentiated osteoblasts. The functional role of miR-15b in osteoblast differentiation was determined using miR-15b mimic/inhibitor and the expression of osteoblast differentiation marker genes such as alkaline phosphatase (ALP), type I collagen genes was decreased by miR-15b inhibitor. Runx2, a bone specific transcription factor is generally required for expression of osteoblast differentiation marker genes and in response to miR-15b inhibitor treatment, Runx2 mRNA expression was not changed; whereas its protein expression was decreased. Even though Smurf1 (SMAD specific E3 ubiquitin protein ligase 1), HDAC4 (histone deacetylase 4), Smad7, and Crim1 were found to be few of miR-15b's putative target genes, there was increased expression of only Smurf1 gene at mRNA and protein levels by miR-15b inhibitor. miR-15b mimic treatment significantly increased and decreased expressions of Runx2 and Smurf1 proteins, respectively. We further identified that the Smurf1 3’UTR is directly targeted by miR-15b using the luciferase reporter gene system. This is well documented that Smurf1 interacts with Runx2 and degrades it by proteasomal pathway. Hence, based on our results we suggest that miR-15b promotes osteoblast differentiation by indirectly protecting Runx2 protein from Smurf1 mediated degradation. Thus, this study identified that miR-15b can act as a positive regulator for osteoblast differentiation.

Description: Robot in Laboratory. Copyright: Max Tactic – Fotolia.com.

Description: Expression of smooth muscle alpha-actin (SMαA) is essential for myofibroblast-mediated wound contraction following tissue injury. The Pur α/β and YB-1 transcriptional repressors govern the DNA-binding activity of serum response factor (SRF) and phosphorylated Smad3 (pSmad3) transcriptional activators during induction of SMαA gene expression in human pulmonary myofibroblasts. In quiescent fibroblasts, Pur α exhibited a novel function in enhancing stability of pre-existing SRF complexes with SMαA core promoter DNA whereas Pur β was more effective in disrupting SRF-DNA interaction. Pur proteins were less efficient competitors of pre-existing, core-promoter complexes containing both SRF and pSmad3 in nuclear extracts from TGFβ1-activated myofibroblasts. TGFβ1 signaling dissociated a SRF/Pur protein complex with concurrent formation of a transient pSmad3/MRTF-A/Pur β complex during early phase myofibroblast differentiation. Pur β was replaced by Pur α in the pSmad3/MRTF-A complex in mature myofibroblasts. Combining all three repressors potently inhibited SRF and pSmad3 binding to promoter DNA in quiescent fibroblasts and TGFβ1-activated myofibroblasts, respectively. The results point to dynamic interplay between transcriptional activators and repressors in regulating SMαA gene output during myofibroblast differentiation. Therapeutic targeting of nucleoprotein complexes regulating the SMαA promoter may prevent excessive myofibroblast accumulation associated with chronic cardiopulmonary fibrosis and dysfunctional tissue remodeling.

Description: The aim of this study was to investigate if chemically produced nanotopography on titanium (Ti) surface induces osteoblast differentiation of cultured human bone marrow mesenchymal stem cells (hMSCs) by regulating the expression of microRNAs (miRs). It was demonstrated that Ti with nanotopography induces osteoblast differentiation of hMSCs as evidenced by upregulation of osteoblast specific markers compared with untreated (control) Ti at day 4. At this time-point, miR-sequencing analysis revealed that 20 miRs were upregulated (〉twofold) while 20 miRs were downregulated (〉threefold) in hMSCs grown on Ti with nanotopography compared with control Ti. Three miRs, namely miR-4448, -4708, and -4773, which were significantly downregulated (〉fivefold) by Ti with nanotopography affect osteoblast differentiation of hMSCs. These miRs that directly target SMAD1 and SMAD4, both key transducers of the bone morphogenetic protein 2 (BMP-2) osteogenic signal, were upregulated by Ti with nanotopography. Overexpression of miR-4448, -4708, and 4773 in MC3T3-E1 pre-osteoblasts noticeably inhibited gene and protein expression of SMAD1 and SMAD4 and therefore repressed the gene expression of key bone markers. Additionally, it was observed that the treatment with BMP-2 displayed a higher osteogenic effect on MC3T3-E1 cells grown on Ti with nanotopography compared with control Ti, suggesting that the BMP-2 signaling pathway was more effective on this surface. Taken together, these results indicate that a complex regulatory network involving a miR-SMAD-BMP-2 circuit governs the osteoblast differentiation induced by Ti with nanotopography. J. Cell. Physiol. © 2014 Wiley Periodicals, Inc.

Description: The most common cause of dilated cardiomyopathy and heart failure (HF) is ischemic heart disease; however, in a third of all patients the cause remains undefined and patients are diagnosed as having idiopathic dilated cardiomyopathy (IDC). Recent studies suggest that many patients with IDC have a family history of HF and rare genetic variants in over 35 genes have been shown to be causative of disease. We employed whole-exome sequencing to identify the causative variant in a large family with autosomal dominant transmission of dilated cardiomyopathy. Sequencing and subsequent informatics revealed a novel 10-nucleotide deletion in the BCL2-associated athanogene 3 ( BAG3 ) gene ((Ch10:del 121436332_12143641: del. 1266_1275 [NM 004281]) that segregated with all affected individuals. The deletion predicted a shift in the reading frame with the resultant deletion of 135 amino acids from the C-terminal end of the protein. Consistent with genetic variants in genes encoding other sarcomeric proteins there was a considerable amount of genetic heterogeneity in the affected family members. Interestingly, we also found that the levels of BAG3 protein were significantly reduced in the hearts from unrelated patients with end-stage HF undergoing cardiac transplantation when compared with non-failing controls. Diminished levels of BAG3 protein may be associated with both familial and non-familial forms of dilated cardiomyopathy. J. Cell. Physiol. © 2014 Wiley Periodicals, Inc.

Description: Cardiosphere-derived cells (CDCs) were cultured from human, murine and rat hearts. Diluted supernatant (conditioned-medium) of the cultures improved the contractile behavior of isolated rat cardiomyocytes (CMCs). This effect is mediated by the paracrine release of cytokines. The present study tested the hypothesis, that the cardiovascular state of the donor‘s heart influences this effect on CMCs and tries to identify the responsible factors. CDCs were cultured from human tissue samples of cardiac surgery and from murine and rat hearts. The supernatants of cultured CDCs from hypertensive humans and rats showed a higher improvement of the contractile behavior of CMCs compared to CDCs of normotensive origin. Subsequently, the cytokine profile of the supernatants was analyzed. Among the cytokines elevated in supernatants originating from hypertensive humans or rats was Interleukin-6. CDCs were also generated from Interleukin-6 -/- -mice and their wildtype littermates. The supernatant of the cultured Interleukin-6 -/- -CDCs had no effect on the contractile behavior, whereas the supernatant of the Interleukin-6 +/+ -CDCs showed a positive effect. To confirm the hypothesis that Interleukin-6 contributes to the paracrine effects, CMCs were incubated with Interleukin-6. It improved the contractile function in a concentration dependent way. Finally, the effect of the supernatant of cultured CDCs derived from a hypertensive human sample could be abolished by simultaneous incubation with a specific Interleukin-6 antibody. CDCs release cytokines that improve the contractile behavior of CMCs. This effect is more intense in CDCs from hypertensive donors. Interleukin-6 is involved in this phenomenon. J. Cell. Physiol. © 2014 Wiley Periodicals, Inc.

Description: Recent technological advances allow for high throughput profiling of biological systems in a cost-efficient manner. The low cost of data generation is leading us to the “big data” era. The availability of big data provides unprecedented opportunities, but it also brings out challenges in data mining and analysis. In this review, we introduce key concepts in the analysis of big data, including both “machine learning” algorithms as well as “unsupervised” and “supervised” examples of each. We note packages for the R programming language that are available perform machine learning analyses. In addition to programming based solutions, we review webservers that allow users with limited or no programming background to perform these analyses on large data compendia. J. Cell. Physiol. © 2014 Wiley Periodicals, Inc.

Description: Polyethersulfone (PES) was blended with poly( D,L -lactide) (PDLLA) to prepare asymmetric membranes using the phase inversion method. The effects of the blend ratios and poly(ethylene glycol) (PEG) as additive on the membrane structure, properties, and performance were investigated. The membranes were characterized by contact angle determination, scanning electron microscopy, porosity measurement, thermogravimetric analysis, degradation tests in compost, and dynamic tests for pure water flux and bakery product wastewater treatment. PES and PEG slightly reduced the membrane contact angle. Increasing the PDLLA concentration in the blend membranes enhanced the membrane degradation in compost and also the membrane porosity. The permeate flux of the membranes was improved, but the rejection of pollution indices did not change noticeably. Porous membranes have been widely applied in the fields of dialysis, clarification, and purification. The composition of membranes prepared by blending polyethersulfone with poly( D,L -lactide) (PDLLA) affected their characteristics. Increases in the concentrations of PDLLA and the additive poly(ethylene glycol) led to more porous structures, causing higher fluxes.

Description: Endometriosis is estimated to affect 10% of women during the reproductive years. The lack of a non-invasive diagnostic test significantly contributes to the long delay between onset of the symptoms and definitive diagnosis of endometriosis. This case–control study was conducted to identify specific endometriosis antigens using 2D gel analysis in women with endometriosis (n = 5) and without endometriosis (n = 5). Differentially expresses spots were analyzed using matrix-assisted laser desorption/ionization-time-of-flight/mass spectrometry (nanoLC-ESI-MS/MS) with MASCOT analysis, in order to identify the corresponding proteins. ELISAs were performed on a different cohort of endometriosis (n = 120) and healthy patients (n = 20) in order to confirm the differential expression of the identified proteins. ROC analysis of ELISA results confirmed the statistical significance of the differential expression for one of these proteins: Zn-alpha2-glycoprotein ( P  = 0.019). We propose the analysis of the expression level of this protein in the serum as a new non-invasive diagnostic test for endometriosis. J. Cell. Physiol. © 2014 Wiley Periodicals, Inc.

Description: The separation efficiency of a pilot-scale zigzag apparatus is investigated numerically using computational fluid dynamics simulations and discrete particle modeling in a coupled manner. The effects of various process variables, like particle size and air flow velocity, and of turbulence models were analyzed. The resulting changes concerning the process performance expressed by separation function and sharpness are discussed. Moreover, the residence time distribution was found to differ for fine and coarse particle discharges. Small particles are easily carried away by the fluid and respond immediately to almost every change in flow velocity. Therefore, they are affected by vortices, which increase their residence times compared to bigger particles. Zigzag air classifiers are superior to simple sieves for the separation of particles and are used in many industrial processes. The separation efficiency of a pilot-scale zigzag apparatus is studied using combined computational fluid dynamics simulations and discrete particle modeling. The process performance is discussed with respect to the chosen modeling approach and the operating conditions.

Description: Although combination chemotherapy and radiotherapy have become the standard of care in numerous tumors, the mechanisms of interaction are often still unclear. The purpose of this study was to analyze the efficacy of radiation treatment and cisplatin sequences and to investigate their mechanisms of interaction. Three melanoma cell lines were used to evaluate in vitro radiation-induced cytotoxicity before and after cisplatin treatment. Expression levels of a panel of genes were determined by real-time RT-PCR. Cytotoxic effect was evaluated by flow cytometry analysis and Comet assay. We also used normal human dermal fibroblasts (HUDE) to evaluate the cytotoxicity of the two treatments by clonogenic assay. Radiation and cisplatin used singly were not particularly effective in reducing proliferation in melanoma cells. Conversely, radiation treatment followed by cisplatin showed a strong synergistic interaction in all cell lines, with a ratio index ranging from 16 to 〉100. The synergistic effect was accompanied by apoptosis induction (up to 40%) and an increase in the percentage of comet-shaped nucleoids from 85% to 99%. In parallel, our results also showed that radiation treatment of HUDE fibroblasts followed by cisplatin only induced weak cytotoxicity. Our findings highlight the efficacy of the sequence radiation → cisplatin in reducing cell proliferation and in inducing apoptosis in melanoma cell lines. This sequence also modulated a network of proteins involved in DNA damage repair. J. Cell. Physiol. © 2014 Wiley Periodicals, Inc.

Description: The thickness of epicardial adipose tissue (EAT), which is an inflammatory source for coronary artery disease (CAD), correlates with insulin resistance. One trigger factor is impaired adipogenesis. Here, our aim was to clarify the underlying mechanisms of insulin resistance on EAT-mesenchymal cells (MC). EAT and subcutaneous adipose tissue (SAT) were collected from 19 patients who were undergoing heart surgery. Their dedifferentiated adipocytes (DAs) and/or MCs were cultured. After the induction of adipogenesis or stimulation with insulin, the expression of adipokines was analyzed using real-time polymerase chain reaction (PCR). Colorimetric assays were performed to measure glucose levels and proliferation rate. Proteins modifications were detected via the proteomic approach and Western blot. Our results showed lower adipogenic ability in EAT-MCs than in SAT-MCs. Maximum adiponectin levels were reached within 28–35 days of exposure to adipogenic inducers. Moreover, the adipogenesis profile in EAT-MCs was dependent on the patients' clinical characteristics. The low adipogenic ability of EAT-MCs might be associated with an insulin-resistant state because chronic insulin treatment reduced the inflammatory cytokine expression levels, improved the glucose consumption, and increased the post-translational modifications (PTMs) of the glycolytic enzyme phosphoglycerate mutase 1 (PGAM1). We found lower adipogenic ability in EAT-MCs than in SAT-MCs. This lower ability level was dependent on gender and the presence of diabetes, obesity, and CAD. Low adipogenesis ability and insulin resistance in EAT-MCs might shed light on the association between EAT dysfunction and cardiovascular disease. J. Cell. Physiol. © 2014 Wiley Periodicals, Inc.

Description: Myocardial infarction (MI) is defined as cardiac cell death due to prolonged ischemia. Although necrotic cell death was considered to be solely responsible for myocyte death during MI, it was recently revealed that apoptosis also plays its part in this death process. Our laboratory has recently shown that endoplasmic reticulum (ER) stress-induced apoptosis is the predominant route for apoptosis during MI and the conventional mitochondrial pathway is bypassed by activation of a small heat shock protein α-crystallin B (CRYAB). Since CRYAB is a direct target of P38 mitogen-activated protein kinase (MAPK) cascade, we were prompted to check the role of P38 MAPK in 20-week-old male Wister rats immediately after infarct formation. Interestingly, parallel activation of mitochondrial apoptotic pathway with an increase in ER stress-induced apoptotic load was observed along with decreased activation of CRYAB and Nrf2 (a pro-survival protein activated in response to ER stress) in MI rats treated with SB203580, a specific inhibitor of P38α and P38β compared to the MI alone. As a cumulative effect, this inhibitor treatment also resulted in significant increase in the levels of caspase3 activity and TUNEL positivity, the end point apoptotic markers. Furthermore, SB203580-treated hypoxic adult cardiomyocytes showed formation of desmin aggregates which were previously associated with impaired cardiac function. Thus, this study shows for the first time the precise mechanism by which P38 MAPK plays a pro-survival role and confers protection of cardiomyocytes, during infarct formation. J. Cell. Physiol. © 2014 Wiley Periodicals, Inc.

Description: Colon cancer remains one of the lethal malignancies in the world. Aberrant activation of canonical Wnt/β-catenin signaling pathway has been observed in colon cancer. In contrast, the non-canonical Wnt signaling functions remain obscure. Wnt5a is a representative non-canonical Wnt ligand which has gained extensive attention nowadays. Wnt5a has been shown to play an important role in EMT in prostate cancer and melanoma, but its role in colon cancer is still ambiguous. Here we have evaluated Wnt5a expression in a large cohort of 217 colon cancers by immunohistochemistry and analyzed its correlation with clinicopathologic characteristics. We found that expression of Wnt5a was diminished significantly in majority of primary colon cancers and negatively related with EMT biomarkers. To further enlighten the mechanism which Wnt5a regulates EMT in vitro , we established ectopic Wnt5a expression models. Protein analysis demonstrated that Wnt5a inhibited EMT and antagonized canonical Wnt signaling in colon cancer cells. Overexpression of Wnt5a impaired cell motility and invasion and inhibited cell proliferation by manipulating Bax. Moreover, Wnt5a suppressed the tumor growth in nude mice and impaired tumorigenicity in vivo . Wnt5a also induced intracellular calcium and activated non-canonical Wnt/Ca 2+ signaling in colon cancer. In summary, although Wnt5a was down-regulated in majority of colon cancers, enhanced Wnt5a expression predict preferable outcome in colon cancer patients. Our findings indicate that Wnt5a might act as tumor suppressor by inhibiting cell proliferation and attenuating EMT in colon cancer cells. Wnt5a could be used as a novel prognostic marker and/or therapeutic target for colon cancer in the future. J. Cell. Physiol. © 2014 Wiley Periodicals, Inc.

Description: In order to observe the impact of different water compositions on sludge dewaterability, assessments of floc sizes using a particle size analyzer and of sludge dewaterability based on the capillary suction time (CST) test were carried out. Synthetic raw water had small floc sizes, and synthetic domestic wastewater had both larger median floc sizes and a better correlation between sludge dewaterability and median floc sizes. The floc size distribution results showed that synthetic raw water is associated with a narrow particle size distribution. In comparison, synthetic domestic wastewater produced a wider distribution. However, the CST values were similar for both waters. Compared to synthetic wastewater, natural wastewater had the largest distribution with generally larger particle sizes. In order to observe the impact of different water compositions on sludge dewaterability, assessments of floc sizes and sludge dewaterability tests were carried out. The particle distribution results show that synthetic raw water is associated with a very narrow range of particles. In comparison, synthetic domestic wastewater produced a larger distribution. The capillary suction times were similar.

Description: The plasma level of the regulatory metabolite adenosine increases during the activation of coagulation and inflammation. Here we investigated the effect of adenosine on modulation of thrombin-mediated proinflammatory responses in HUVECs. We found that adenosine inhibits the barrier-disruptive effect of thrombin in HUVECs by a concentration-dependent manner. Analysis of cell surface expression of adenosine receptors revealed that A 2A and A 2B are expressed at the highest level among the four receptor subtypes (A 2B  〉 A 2A  〉 A 1  〉 A 3 ) on HUVECs. The barrier-protective effect of adenosine in response to thrombin was recapitulated by the A 2A specific agonist, CGS 21680, and abrogated both by the siRNA knockdown of the A 2A receptor and by the A 2A -specific antagonists, ZM-241385 and SCH-58261. The thrombin-induced RhoA activation and its membrane translocation were both inhibited by adenosine in a cAMP-dependent manner, providing a molecular mechanism through which adenosine exerts a barrier-protective function. Adenosine also inhibited thrombin-mediated activation of NF-κB and decreased adhesion of monocytic THP-1 cells to stimulated HUVECs via down-regulation of expression of cell surface adhesion molecules, VCAM-1, ICAM-1, and E-selectin. Moreover, adenosine inhibited thrombin-induced elevated expression of proinflammatory cytokines, IL-6 and HMGB-1; and chemokines, MCP-1, CXCL-1, and CXCL-3. Taken together, these results suggest that adenosine may inhibit thrombin-mediated proinflammatory signaling responses, thereby protecting the endothelium from injury during activation of coagulation and inflammation. J. Cell. Physiol. © 2014 Wiley Periodicals, Inc.

Description: Dental pulp originating from the neural crest is considered a better source of postnatal stem cells for cell-based therapies in neurodegenerative diseases. Dental Pulp Stem Cells (DPSCs) have been shown to differentiate into cell-types of cranial neural crest ontology; however, their ability to differentiate to functional neurons of the central nervous system remains to be studied. We hypothesized that midbrain cues might commit DPSCs to differentiate to functional dopaminergic cell-type. As expected, DPSCs in their naïve state spontaneously expressed early and mature neuronal markers like nestin, musashi12, β tubulin III and Map2ab. On exposure to midbrain cues (sonic hedgehog, fibroblast growth factor 8 and basic fibroblast growth factor), DPSCs showed upregulation of dopaminergic neuron-specific transcription factors Nuclear Receptor related protein 1 (Nurr1), Engrailed 1 (En1) and paired-like homeodomain transcription factor 3 (Pitx3) as revealed by real-time RT-PCR. Immunofluorescence and flow cytometry analysis showed enhanced expression of mature neuronal marker Map2ab and dopaminergic-neuronal markers [tyrosine hydroxylase (TH), En1, Nurr1 and Pitx3], with nearly 77% of the induced DPSCs positive for TH. Functional studies indicated that the induced DPSCs could secrete dopamine constitutively and upon stimulation with potassium chloride (KCl) and adenosine triphosphate (ATP), as measured by dopamine ELISA. Additionally, the induced DPSCs showed intracellular Ca 2+ influx in the presence of KCl, unlike control DPSCs. ATP-stimulated Ca 2+ influx was observed in control and induced DPSCs, but only the induced cells secreted dopamine. Our data clearly demonstrates for the first time that DPSCs in the presence of embryonic midbrain cues show efficient propensity towards functional dopaminergic cell-type. J. Cell. Physiol. © 2014 Wiley Periodicals, Inc.

Description: The direct-ignition technology of an entrained-flow coal-water slurry gasifier with hot-oxygen burner (HOB) is presented. Experimental tests were carried out using diesel oil and coal-water slurry in a bench-scale entrained-flow opposed multi-burner (OMB) gasifier. Using a CCD camera equipped with high-temperature endoscope, flame images inside the gasifier were recorded and subsequently processed. The results show that the HOB diesel flame is more stable than the normal two-channel burner flame typically used. Entrained-flow coal-water slurry gasifiers can start up using the HOB directly without a higher preheated wall temperature. An HOB can potentially save gasifier preheating time and operation costs and enables a wide range of future applications. Entrained-flow gasification offers one of the cleanest ways to obtain energy from coal. The direct-ignition technology of an entrained-flow coal-water slurry gasifier with a new type of hot-oxygen burner (HOB) was extensively studied to prove the feasibility of the gasifier startup with this setup. The HOB permits to effectively decrease the gasifier preheating time and the system operation costs.

Description: Different approaches were analyzed to combine the tuneable micropore structure of carbide-derived carbons with a foam-like secondary porosity. The resulting structured catalyst supports were characterized in detail and applied in the model reaction of ethene hydrogenation. Preparation methods studied were dip-coating using polytetrafluoroethylene as binder on cellular metal structures, a chemical vapor deposition coating of the metal structures with thin carbide layers and subsequent conversion to carbide-derived carbon, and the partial or full conversion of carbide foams to carbon/carbide composites. For the binder method, optimal parameters for stable slurry preparation as well as for calcination of the slurry were obtained. It could further be demonstrated that the conversion of carbide foams into carbon/carbide composites leads to an appreciation between decreasing mechanical strength and increasing specific surface area. Structured catalysts are advantageous to conventional fixed-bed systems. Carbide-derived carbons with tuneable microporous structure were combined with a foamlike catalyst structure. The three preparation methods studied resulted in highly active catalysts. Optimal conditions for the easy-to-apply dip-coating method to stabilize the slurry for successful coating were determined and evaluated.

Description: An opposed multi-burner (OMB) entrained-flow gasifier with coal water slurry feeding is developed by the East China University of Science and Technology. A 3D model is employed to numerically simulate the gas flow field, motion of char particles, and distributions of temperature and gaseous components in an OMB gasifier and in a conceptual two-stage gasifier modified from the OMB gasifier (TS-OMB gasifier). Results show that the TS-OMB gasifier produces higher concentration and productivity of the effective gases (CO+H 2 ) with a slightly higher carbon conversion than the OMB gasifier. The reasons for the differences between these two types of gasifier are discussed by means of numerical simulation. This information is valuable for guiding the design of an advanced OMB gasifier. With an increasing demand for syngas, entrained-flow gasifiers for coal conversion are installed at the fastest pace due to advantages such as low pollutant emission. Simulations of the gas component distribution, amongst other key factors, in an opposed multi-burner (OMB) gasifier and modified two-stage OMB gasifier were compared. The results are valuable for future design of an advanced OMB gasifier.

Description: The chromatin remodeling complex SWI/SNF and the transcription factor C/EBPβ play critical roles in osteoblastic cells as they jointly control transcription of a number of bone-related target genes. The largest C/EBPβ isoform, LAP*, possesses a short additional N-terminal domain that has been proposed to mediate the interaction of this factor with SWI/SNF in myeloid cells. Here we examine the requirement of a functional N-terminus in C/EBPβ-LAP* for binding SWI/SNF and for recruiting this complex to the Ric-8B gene to mediate transcriptional repression. We find that both C/EBPβ-LAP* and SWI/SNF simultaneously bind to the Ric-8B promoter in differentiating osteoblasts that repress Ric-8B expression. This decreased expression of Ric-8B is not accompanied by significant changes in histone acetylation at the Ric-8B gene promoter sequence. A single aminoacid change at the C/EBPβ-LAP* N-terminus (R3L) that inhibits C/EBPβ-LAP*-SWI/SNF interaction, also prevents SWI/SNF recruitment to the Ric-8B promoter as well as C/EBPβ-LAP*-dependent repression of the Ric-8B gene. Inducible expression of the C/EBPβ-LAP*R3L protein in stably transfected osteoblastic cells demonstrates that this mutant protein binds to C/EBPβ-LAP*-target promoters and competes with the endogenous C/EBPβ factor. Together our results indicate that a functional N-terminus in C/EBPβ-LAP* is required for interacting with SWI/SNF and for Ric-8B gene repression in osteoblasts. J. Cell. Physiol. 9999: XX–XX, 2014. © 2014 Wiley Periodicals, Inc.

Description: Despite a regain of interest recently in ERK3 kinase signaling, the molecular regulations of both ERK3 gene expression and protein kinase activity are still largely unknown. While it is shown that disruption of ERK3 gene causes neonatal lethality, cell type-specific functions of ERK3 signaling remain to be explored. In this study, we report that ERK3 gene expression is upregulated by cytokines through c-Jun in endothelial cells; c-Jun binds to the ERK3 gene and regulates its transcription. We further reveal a new role for ERK3 in regulating endothelial cell migration, proliferation and tube formation by upregulating SRC-3/SP-1-mediated VEGFR2 expression. The underlying molecular mechanism involves ERK3-stimulated formation of a transcriptional complex involving coactivator SRC-3, transcription factor SP-1 and the secondary coactivator CBP. Taken together, our study identified a molecular regulatory mechanism of ERK3 gene expression and revealed a previously unknown role of ERK3 in regulating endothelial cell functions. J. Cell. Physiol. 9999: XX–XX, 2014. © 2014 Wiley Periodicals, Inc.

Description: Core Binding Factor β (CBFβ) is complexed with the RUNX family of transcription factors in the nucleus to support activation or repression of genes related to bone (RUNX2), hematopoiesis (RUNX1) and gastrointestinal (RUNX3) development. Furthermore, RUNX proteins contribute to the onset and progression of different types of cancer. Although CBFβ localizes to cytoskeletal architecture, its biological role in the cytoplasmic compartment remains to be established. Additionally, the function and localization of CBFβ during the cell cycle are important questions relevant to its biological role. Here we show that CBFβ dynamically distributes in different stages of cell division and importantly is present during telophase at the midbody, a temporal structure important for successful cytokinesis. A functional role for CBFβ localization at the midbody is supported by striking defects in cytokinesis that include polyploidy and abscission failure following siRNA-mediated downregulation of endogenous CBFβ or overexpression of the inv(16) fusion protein CBFβ-SMMHC. Our results suggest that CBFβ retention in the midbody during cytokinesis reflects a novel function that contributes to epigenetic control. J. Cell. Physiol. 9999: XX–XX, 2014. © 2014 Wiley Periodicals, Inc.

Description: Factory. Copyright: Dmitry Lavrenyuk – Fotolia.com .

Description: Our previous work showed that the expression of bone morphogenetic protein-4 (BMP4) was up-regulated in pathological cardiac hypertrophy models and BMP4 induced cardiomyocyte hypertrophy and apoptosis. Bone morphogenetic protein-2 (BMP2) and BMP4 share greater than 80% amino acid homology and there exists an interaction between BMP2 and BMP4, so the aim of the present study was to elucidate the changes of BMP2 in the cardiac hypertrophy models and the effects of BMP2 on BMP4-induced cardiomyocyte hypertrophy and apoptosis. The in vivo cardiac hypertrophy models were induced by pressure-overload and swimming exercise in mice. BMP2 mRNA and protein expressions increased in pressure-overload and swimming-exercise induced cardiac hypertrophy. BMP2 itself did not elicit cardiomyocyte hypertrophy and apoptosis, but antagonized BMP4-induced cardiomyocyte hypertrophy and apoptosis. BMP2 stimulated Akt in cardiomyocytes and Akt inhibitor prevented the antagonism of BMP2 on BMP4-induced cardiomyocyte apoptosis. Furthermore, BMP2 inhibited BMP4-induced JNK activation in cardiomyocytes. In conclusion, BMP2 antagonizes BMP4-induced cardiomyocyte hypertrophy and apoptosis. The anti-apoptotic effects of BMP2 on BMP4-induced cardiomyocyte apoptosis might be through activating Akt and inhibiting JNK activation. J. Cell. Physiol. 9999: XX–XX, 2014. © 2014 Wiley Periodicals, Inc.

Description: Expression studies have implicated FLRT2 in cranial neural crest cell migration and prechondrogenic cell condensation during craniofacial skeletogenesis. We aimed to determine whether FLRT2 was involved in mediating cell-matrix interactions in the ATDC5 chondroprogenitor cell line. Immunolocalization experiments of ATDC5 cells revealed that FLRT2 was present on the cell membrane as well as extracellularly, where it colocalized with Fibronectin (Fn). After cell extraction of the matrix, FLRT2 was identified in the ATDC5-derived extracellular matrix (ECM) and was further found to be associated with Fn-coated beads in cell cultures. Blockage of Fn fibril formation via a blocking peptide resulted in a concomitant decrease in extracellular FLRT2 accumulation. Over a 7-day period following the replenishment of the Fn blocking peptide to the cultures, there was a partial rebound in Fn fibril formation that was accompanied by a concomitant reappearance of FLRT2 co-expression. Co-immunoprecipitation confirmed that FLRT2 and Fn interacted, either directly or indirectly. Immunoprecipitation and Western blot analyses with antibodies recognizing epitopes located on the extra- and intracellular domains of FLRT2 further revealed the presence of different sized bands, suggesting that FLRT2 may exist in both membrane-bound and shed forms. Our data therefore provide evidence that FLRT2 and/or its cleavage products may be cooperating with Fn and other ECM proteins to regulate critical cellular events. Further studies will be necessary in delineate more precisely the roles of FLRT2 in mediating cell- and cell-matrix interactions during normal development. J. Cell. Physiol. 9999: XX–XX, 2014. © 2014 Wiley Periodicals, Inc.

Description: Grb10 is an intracellular adaptor protein which binds directly to several growth factor receptors, including those for insulin and insulin-like growth factor receptor-1 (IGF-1), and negatively regulates their actions. Grb10-ablated (Grb10 -/- ) mice exhibit improved whole body glucose homeostasis and an increase in muscle mass associated specifically with an increase in myofiber number. This suggests that Grb10 may act as a negative regulator of myogenesis. In this study, we investigated in vitro , the molecular mechanisms underlying the increase in muscle mass and the improved glucose metabolism. Primary muscle cells isolated from Grb10 -/- mice exhibited increased rates of proliferation and differentiation compared to primary cells isolated from wild-type mice. The improved proliferation capacity was associated with an enhanced phosphorylation of Akt and ERK in the basal state and changes in the expression of key cell cycle progression markers involved in regulating transition of cells from the G1 to S phase (e.g., retinoblastoma (Rb) and p21). The absence of Grb10 also promoted a faster transition to a myogenin positive, differentiated state. Glucose uptake was higher in Grb10 -/- primary myotubes in the basal state and was associated with enhanced insulin signaling and an increase in GLUT4 translocation to the plasma membrane. These data demonstrate an important role for Grb10 as a link between muscle growth and metabolism with therapeutic implications for diseases, such as muscle wasting and type 2 diabetes. J. Cell. Physiol. © 2014 Wiley Periodicals, Inc.

Description: Age-related bone loss is characterized by reduced osteoblastogenesis and excessive bone marrow adipogenesis. The mechanisms governing bone marrow mesenchymal stromal cell (BMSC) differentiation into adipocytes or osteoblasts during aging are unknown. We show here that overexpressing N-cadherin ( Cadh2 ) in osteoblasts increased BMSC adipocyte differentiation and reduced osteoblast differentiation in young transgenic (Tg) mice whereas this phenotype was fully reversed with aging. The reversed phenotype with age was associated with enhanced Wnt5a and Wnt10b expression in osteoblasts and a concomitant increase in BMSC osteogenic differentiation. Consistent with this mechanism, conditioned media from young wild type osteoblasts inhibited adipogenesis and promoted osteoblast differentiation in BMSC from old Cadh2 Tg mice, and this response was abolished by Wnt5a and Wnt10b silencing. Transplantation of BMSC from old Cadh2 Tg mice into young Tg recipients increased Wnt5a and Wnt10b expression and rescued BMSC osteogenic differentiation. In senescent osteopenic mice, blocking the CADH2–Wnt interaction using an antagonist peptide increased Wnt5a and Wnt10b expression, bone formation, and bone mass. The data indicate that Cadh2/Wnt interaction in osteoblasts regulates BMSC lineage determination, bone formation, and bone mass and suggest a therapeutic target for promoting bone formation in the aging skeleton. J. Cell. Physiol. © 2014 Wiley Periodicals, Inc.

Description: Aging-related chronic illness is a price we have to pay to live longer. Prevalent among the oldest old, the condition limits their functional independence and also aggravates the course of several age-related chronic diseases. Thus, the search is on for efficient therapies that will mitigate age-related pathologies. In this article, we point out the potential clinical implications of recent provocative basic research in the field. New possible targets have been recently discovered, are clearly involved in age-related pathologies and might benefit the treatment of other age-related conditions, particularly metabolic diseases. J. Cell. Physiol. © 2014 Wiley Periodicals, Inc.

Description: Nemo-like kinase (Nlk) is related to the mitogen-activated protein (MAP) kinases and known to regulate signaling pathways involved in osteoblastogenesis. In vitro Nlk suppresses osteoblastogenesis, but the consequences of the Nlk inactivation in the skeleton in vivo are unknown. To study the function of Nlk, Nlk loxP/loxP mice, where the Nlk exon2 is flanked by lox P sequences, were mated with mice expressing the Cre recombinase under the control of the paired-related homeobox gene 1 ( Prx1 ) enhancer ( Prx1-Cre ), the Osterix ( Osx-Cre ) or the osteocalcin/bone gamma carboxyglutamate protein ( Bglap-Cre ) promoter. Prx1-Cre;Nlk Δ/Δ mice did not exhibit a skeletal phenotype except for a modest increase in trabecular number and connectivity observed only in 3-month-old male mice. Osx-Cre;Nlk Δ/Δ male and female mice exhibited an increase in trabecular bone volume secondary to an increased trabecular number at 3 months of age. Bone histomorphometry revealed a decrease in osteoclast number and eroded surface in male mice, and decreased osteoblast number and function in female mice. Expression of osteoprotegerin mRNA was increased in calvarial extracts, explaining the decreased osteoclast and osteoblast number. The conditional deletion of Nlk in mature osteoblasts ( Bglap-Cre;Nlk Δ/Δ ) resulted in no skeletal phenotype in 1- to 6-month-old male or female mice. In conclusion, when expressed in undifferentiated osteoblasts, Nlk is a negative regulator of skeletal homeostasis possibly by targeting signals that regulate osteoclastogenesis and bone resorption. J. Cell. Physiol. © 2014 Wiley Periodicals, Inc.

Description: Pentraxin-3 (PTX3), also known as tumor necrosis factor-stimulated gene 14 (TSG-14), is produced by immune and vascular cells in response to pro-inflammatory signals and is therefore a multipotent inflammatory mediator. The present study showed that during human osteoblast (OB) differentiation, precursor OBs (pOBs), but not mature OB, highly expressed PTX3. TNFα treatment elevated the PTX3 expression of pOBs. When mice were injected with lipopolysaccharide, which induces an inflammatory osteolytic condition characterized by trabecular bone destruction and high osteoclastogenesis, their bone marrow cells expressed elevated levels of PTX3 protein. Exogenous PTX3 did not directly affect osteoclast (OC) or OB differentiation. However, when pOBs and precursor OCs were co-cultured, exogenous PTX3 significantly increased the number of tartrate-resistant acid phosphatase-positive multinucleated cells (i.e., OC cells) by increasing the pOB mRNA expression and protein secretion of RANK ligand (RANKL). This was accompanied with increased Runt-related transcription factor 2 (Runx2) expression in the pOBs. Knock-down of endogenous PTX3 with small-interfering RNA did not change the osteogenic potential of pOBs but suppressed their production of RANKL and reduced osteoclastogenesis. Finally, TNFα treatment of the co-culture elevated PTX3 expression by the pOBs and increased OC formation. This effect was suppressed by PTX3 knock-down by decreasing RANKL expression. Thus, the PTX3-driven increase in the osteoclastogenic potential of pOBs appears to be mediated by the effect of PTX3 on pOB RANKL production. These findings suggest that PTX3 is an inflammatory mediator that contributes to the deteriorating osteolytic condition of inflamed bone. J. Cell. Physiol. © 2014 Wiley Periodicals, Inc.

Description: Steel pipe line. Copyright: supakitmod – Fotolia.com.

Description: Light olefin and paraffin are commonly separated by energy-intensive cryogenic distillation. Membrane/distillation hybrid systems constitute an economical alternative separation process. Different configurations of this hybrid system are studied for olefin-paraffin separation with emphasis on C 3 separation. An approach based on the McCabe-Thiele method is applied to analyze different process configurations. A facilitated transport membrane is considered as membrane type. Both new column design and augmentation of an existing distillation column by a membrane module are considered. Numerical examples are considered for the separation of propane from propylene through different hybridization shapes with facilitated transport membranes. The energy requirement can be halved using hybrid systems. Membrane/distillation hybrid systems represent an economical alternative separation process compared to the commonly applied energy-intensive cryogenic distillation for separation of light olefin and paraffin. Different process configurations of such a hybrid system are evaluated. Under optimum conditions the energy requirement could be halved.

Description: Shear stress secondary to increased pulmonary blood flow (PBF) is elevated in some children born with congenital cardiac abnormalities. However, the majority of these patients do not develop pulmonary edema, despite high levels of permeability inducing factors. Previous studies have suggested that laminar fluid shear stress can enhance pulmonary vascular barrier integrity. However, little is known about the mechanisms by which this occurs. Using microarray analysis, we have previously shown that Sox18, a transcription factor involved in blood vessel development and endothelial barrier integrity, is up-regulated in an ovine model of congenital heart disease with increased PBF (shunt). By subjecting ovine pulmonary arterial endothelial cells (PAEC) to laminar flow (20 dyn/cm 2 ), we identified an increase in trans-endothelial resistance (TER) across the PAEC monolayer that correlated with an increase in Sox18 expression. Further, the TER was also enhanced when Sox18 was over-expressed and attenuated when Sox18 expression was reduced, suggesting that Sox18 maintains the endothelial barrier integrity in response to shear stress. Further, we found that shear stress up-regulates the cellular tight junction protein, Claudin-5, in a Sox18 dependent manner, and Claudin-5 depletion abolished the Sox18 mediated increase in TER in response to shear stress. Finally, utilizing peripheral lung tissue of 4 week old shunt lambs with increased PBF, we found that both Sox18 and Claudin-5 mRNA and protein levels were elevated. In conclusion, these novel findings suggest that increased laminar flow protects endothelial barrier function via Sox18 dependent up-regulation of Claudin-5 expression.

Description: Estrogen deficiency is a major risk factor for osteoporosis that is associated with bone inflammation and resorption. Half of women over the age of 50 will experience an osteoporosis related fracture in their lifetime, thus novel therapies are needed to combat post-menopausal bone loss. Recent studies suggest an important role for gut-bone signaling pathways and the microbiota in regulating bone health. Given that the bacterium Lactobacillus reuteri ATCC PTA 6475 ( L. reuteri ) secretes beneficial immunomodulatory factors, we examined if this candidate probiotic could reduce bone loss associated with estrogen deficiency in an ovariectomized (Ovx) mouse menopausal model. Strikingly, L. reuteri treatment significantly protected Ovx mice from bone loss. Osteoclast bone resorption markers and activators (Trap5 and RANKL) as well as osteoclastogenesis are significantly decreased in L. reuteri treated mice. Consistent with this, L. reuteri suppressed Ovx-induced increases in bone marrow CD4+ T-lymphocytes (which promote osteoclastogenesis) and directly suppressed osteoclastogenesis in vitro. We also identified that L. reuteri treatment modifies microbial communities in the Ovx mouse gut. Together, our studies demonstrate that L. reuteri treatment suppresses bone resorption and loss associated with estrogen deficiency. Thus, L. reuteri treatment may be a straightforward and cost-effective approach to reduce post-menopausal bone loss. J. Cell. Physiol. © 2014 Wiley Periodicals, Inc.

Description: In neurogenerative diseases, comprising Alzheimer's (AD), functional alteration in autophagy is considered one of the pathological hallmarks and a promising therapeutic target. Epidemiological investigations on the possible causes undergoing these diseases have suggested that electromagnetic fields (EMF) exposition can contribute to their etiology. On the other hand, EMF have therapeutic implications in reactivating neuronal functionality. To partly clarify this dualism, the effect of low-frequency EMF (LF-EMF) on the modulation of autophagy was investigated in human neuroblastoma SH-SY5Y cells, which were also subsequently exposed to Aβ peptides, key players in AD. The results primarily point that LF-EMF induce a significant reduction of microRNA 30a (miR-30a) expression with a concomitant increase of Beclin1 transcript ( BECN1 ) and its corresponding protein. Furthermore, LF-EMF counteract the induced miR-30a up-regulation in the same cells transfected with miR-30a mimic precursor molecules and, on the other side, rescue Beclin1 expression after BECN1 siRNA treatment. The expression of autophagy-related markers (ATG7 and LC3B-II) as well as the dynamics of autophagosome formation were also visualized after LF-EMF exposition. Finally, different protocols of repeated LF-EMF treatments were assayed to contrast the effects of Aβ peptides in vitro administration. Overall, this research demonstrates, for the first time, that specific LF-EMF treatments can modulate in vitro the expression of a microRNA sequence, which in turn affects autophagy via Beclin1 expression. Taking into account the pivotal role of autophagy in the clearance of protein aggregates within the cells, our results indicate a potential cytoprotective effect exerted by LF-EMF in neurodegenerative diseases such as AD. J. Cell. Physiol. © 2014 Wiley Periodicals, Inc.

Description: AQP3 has been correlated with higher transport of glycerol, increment of ATP content, and larger proliferation capacity. Recently, we described the gold(III) complex Auphen as a very selective and potent inhibitor of AQP3's glycerol permeability (P gly ). Here we evaluated Auphen effect on the proliferation of various mammalian cell lines differing in AQP3 expression level: no expression (PC12), moderate (NIH/3T3) or high (A431) endogenous expression, cells stably expressing AQP3 (PC12-AQP3), and human HEK293T cells transiently transfected (HEK-AQP3) for AQP3 expression. Proliferation was evaluated in the absence or presence of Auphen (5 μM) by counting number of viable cells and analyzing 5-bromo-2′-deoxyuridine (BrdU) incorporation. Auphen reduced ≈50% the proliferation in A431 and PC12-AQP3, ≈15% in HEK-AQP3 and had no effect in wt-PC12 and NIH/3T3. Strong arrest in the S-G2/M phases of the cell cycle, supported by analysis of cyclins (A, B1, D1, E) levels, was observed in AQP3-expressing cells treated with Auphen. Flow-cytometry of propidium iodide incorporation and measurements of mitochondrial dehydrogenases activity confirmed absence of cytotoxic effect of the drug. Functional studies evidenced ≈50% inhibition of A431 P gly by Auphen, showing that the compound's anti-proliferative effect correlates with its ability to inhibit AQP3 P gly . Role of Cys-40 on AQP3 permeability blockage by Auphen was confirmed by analyzing the mutated protein (AQP3-Ser-40). Accordingly, cells transfected with mutated AQP3 gained resistance to the antiproliferative effect of Auphen. These results highlight an Auphen inhibitory effect on proliferation of cells expressing AQP3 and suggest a targeted therapeutic effect on carcinomas with large AQP3 expression. J. Cell. Physiol. © 2014 Wiley Periodicals, Inc.

Description: There is a critical need for techniques that directly monitor protein synthesis within cells isolated from normal and diseased tissue. Fibrotic disease, for which there is no drug treatment, is characterized by the overexpression of collagens. Here, we use a bioinformatics approach to identify a pair of glycine and proline isoacceptor tRNAs as being specific for the decoding of collagen mRNAs, leading to development of a FRET-based approach, dicodon monitoring of protein synthesis (DiCoMPS), that directly monitors the synthesis of collagen. DiCoMPS aimed at detecting collagen synthesis will be helpful in identifying novel anti-fibrotic compounds in cells derived from patients with fibrosis of any etiology, and, suitably adapted, should be widely applicable in monitoring the synthesis of other proteins in cells. J. Cell. Physiol. © 2014 Wiley Periodicals, Inc.

Description: The proper organization and segregation of chromosomes during cell division is essential to the preservation of genomic integrity. To understand the mechanisms that spatially control the arrangement and dynamics of mitotic chromosomes requires imaging assays to quantitatively resolve their positions and movements. Here, we will discuss analytical approaches to investigate the position-dependent control of mitotic chromosomes in cultured cells. These methods can be used to dissect the specific contributions of mitotic proteins to the molecular control of chromosome dynamics. J. Cell. Physiol. 9999: XX–XX, 2014. © 2014 Wiley Periodicals, Inc.

Description: Hyperinsulinemia is a likely cause of the increased cancer incidence and mortality in diabetic patients, but its role is difficult to define in vivo. Previous in vitro studies testing the mitogenic potential of insulin and its analogs provided incomplete and sometimes contradictory results. To better evaluate cancer cell responsiveness to insulin, to its analogs and to IGF-I, we measured under identical experimental conditions cell proliferation, invasiveness, and foci formation in six cancer cell lines with different insulin receptor family expression levels. The cancer cells studied have a different expression of insulin receptor (IR), its isoforms (IR-A and IR-B), and of the IGF-I receptor. The data indicate that insulin stimulates proliferation in all cancer cell lines, invasiveness in some, and foci formation in none. Cancer cell responses to insulin (and IGF-I) are not related to receptor expression levels; moreover, hormone-stimulated proliferation and invasiveness are not correlated. IGF-I is a more potent stimulator than insulin in most but not all cancer cell lines. Insulin analogs including M1 and M2 Glargine metabolites stimulate cancer cells similar to insulin. However, exceptions occur for specific analogs in particular cancer cells. In conclusion, in vitro insulin is an effective growth factor for all cancer cells but the biological response to insulin cannot be predicted on the basis of receptor expression levels. In the clinical setting, these observations should be taken in account when deciding treatment for diabetic patients who are at risk of undiscovered cancer or survivors of oncological diseases. J. Cell. Physiol. 9999: XX–XX, 2014. © 2014 Wiley Periodicals, Inc.

Description: Despite the importance of the ErbB2/3 heterodimer in breast cancer progression, the negative regulation of these receptors is still poorly understood. We demonstrate here for the first time that the ErbB3/4 ligand heregulin (HRG) reduced both ErbB2 and ErbB3 mRNA and protein levels in human breast cancer cell lines. In contrast, EGFR levels were unaffected by HRG treatment. The effect was rapid with a decline in steady-state mRNA levels first noted 2 h after HRG treatment. HRG reduced the rate of transcription of ErbB2 and ErbB3 mRNA, but did not affect ErbB2 or ErbB3 mRNA stability. To test if ErbB2 kinase activity was required for the HRG-induced downregulation, we treated cells with the ErbB2/EGFR inhibitor lapatinib. Lapatinib diminished the HRG-induced decrease in ErbB2 and ErbB3 mRNA and protein, suggesting that the kinase activity of EGFR/ErbB2 is involved in the HRG-induced receptor downregulation. Further, HRG-mediated decreases in ErbB2/3 mRNA transcription are reversed by inhibiting the AKT but not MAPK pathway. To examine the functional consequences of HRG-mediated decreases in ErbB receptor levels, we performed cell-cycle analysis. HRG blocked cell-cycle progression and lapatinib reversed this block. Our findings support a role for HRG in the negative regulation of ErbB expression and suggest that inhibition of ErbB2/3 signaling by ErbB2 directed therapies may interfere with this process. J. Cell. Physiol. © 2014 Wiley Periodicals, Inc.

Description: Angiotensin II (Ang II), a biologically active peptide of the renin–angiotensin system (RAS), plays an important role in promoting cell migration via Angiotensin II type 1 receptor (AT1R). In this study, we examined the mechanisms by which Ang II affected cell migration in AT1R-positive MDA-MB-231 human breast cancer cells. Ang II increased cell migration and expression of matrix metalloproteinase (MMP)-2,-9 in a dose-dependent manner. Ang II-mediated cell migration was reduced by specific blocking of MMP-2 and MMP-9, as well as with pretreatment with inhibitors of AT1R, phosphatidylinositol 3-kinase (PI3K), Akt, and NF-κB. Similarly, Ang II-mediated expression of MMP-2,-9 was downregulated by pretreatment with inhibitors of AT1R and PI3K. In addition, Ang II treatment significantly induced phosphorylation of PI3K, Akt, and resulted in increased NF-κB activity. These findings suggest that Ang II activates the AT1R/PI3K/Akt pathway, which further activates IKKα/β and NF-κB, resulting in enhanced expression of MMP-2,-9 and migration in human breast cancer cells. Therefore, targeting Ang II/AT1R/PI3K/Akt/NF-κB signaling could be a novel anti-metastatic therapy for breast cancer. J. Cell. Physiol. © 2014 Wiley Periodicals, Inc.

Description: Glioblastoma (GBM) remains the most aggressive and lethal brain tumor due to its molecular heterogeneity and high motility and invasion capabilities of its cells, resulting in high resistance to current standard treatments (surgery, followed by ionizing radiation combined with Temozolomide chemotherapy administration). Locus amplification, gene overexpression, and genetic mutations of epidermal growth factor receptor ( EGFR ) are hallmarks of GBM that can ectopically activate downstream signaling oncogenic cascades such as PI3K/Akt/mTOR pathway. Importantly, alteration of this pathway, involved also in the regulation of autophagy process, can improve radioresistance in GBM cells, thus promoting the aggressive phenotype of this tumor. In this work, the endogenous EGFR expression profile and autophagy were modulated to increase radiosensitivity behavior of human T98G and U373MG GBM cells. Our results primarily indicated that EGFR interfering induced radiosensitivity according to a decrease of the clonogenic capability of the investigated cells, and an effective reduction of the in vitro migratory features. Moreover, EGFR interfering resulted in an increase of Temozolomide (TMZ) cytotoxicity in T98G TMZ-resistant cells. In order to elucidate the involvement of the autophagy process as pro-death or pro-survival role in cells subjected to EGFR interfering, the key autophagic gene ATG7 was silenced, thereby producing a transient block of the autophagy process. This autophagy inhibition rescued clonogenic capability of irradiated and EGFR -silenced T98G cells, suggesting a pro-death autophagy contribution. To further confirm the functional interplay between EGFR and autophagy pathways, Rapamycin-mediated autophagy induction during EGFR modulation promoted further impairment of irradiated cells, in terms of clonogenic and migration capabilities. Taken together, these results might suggest a novel combined EGFR-autophagy modulation strategy, to overcome intrinsic GBM radioresistance, thus improving the efficacy of standard treatments. J. Cell. Physiol. © 2014 Wiley Periodicals, Inc.

Description: Apoptosis in most cell types is accompanied by altered Ca 2+ homeostasis. During apoptosis, caspase-3 mediated cleavage of the type 1 inositol 1,4,5-trisphosphate receptor (IP 3 R1) generates a 95-kDa C-terminal fragment (C-IP 3 R1), which represents the channel domain of the receptor. Aged mouse eggs display abnormal Ca 2+ homeostasis and express C-IP 3 R1, although whether or not C-IP 3 R1 expression contributes to Ca 2+ misregulation or a decrease in developmental competency is unknown. We sought to answer these questions by injecting in mouse oocytes and eggs cRNAs encoding C-IP 3 R1. We found that: (1) expression of C-IP 3 R1 in eggs lowered the Ca 2+ content of the endoplasmic reticulum (ER), although, as C-IP 3 R1 is quickly degraded at this stage, its expression did not impair pre-implantation embryo development; (2) expression of C-IP 3 R1 in eggs enhanced fragmentation associated with aging; (3) endogenous IP 3 R1 is required for aging associated apoptosis, as its down-regulation prevented fragmentation, and expression of C-IP 3 R1 in eggs with downregulated IP 3 R1 partly restored fragmentation; (4) C-IP 3 R1 expression in GV oocytes resulted in persistent levels of protein, which abolished the increase in the ER releasable Ca 2+ pool that occurs during maturation, undermined the Ca 2+ oscillatory ability of matured eggs and their activation potential. Collectively, this study supports a role for IP 3 R1 and C-IP 3 R1 in regulating Ca 2+ homeostasis and the ER Ca 2+ content during oocyte maturation. Nevertheless, the role of C-IP 3 R1 on Ca 2+ homeostasis in aged eggs seems minor, as in MII eggs the majority of endogenous IP 3 R1 remains intact and C-IP 3 R1 undergoes rapid turnover. J. Cell. Physiol. © 2014 Wiley Periodicals, Inc.

Description: Increasing evidence supports the role of epigenetics in the development of autoimmune disorders and the possibility of using epigenetic modifying drugs in the context of MS has not yet been investigated. We have explored the effect of the hypomethylating agent 5-aza-2′-deoxycytidine (DAC) in two murine models of experimental allergic encephalomyelitis (EAE). DAC treatment was associated with a significant amelioration of the clinical and histological hallmarks of EAE in both models. These effects were observed both in prophylactic and therapeutic regimens. The milder course of the disease was associated with a reduction in the number of spinal cord infiltrating lymphocytes and amelioration of the histopathological signs associated with EAE. In addition, increased transcript levels of anti-inflammatory cytokines and decreased mRNA expression of pro-inflammatory mediators were also observed. Finally, DAC treatment increased the percentage of circulating regulatory T cells by inducing Foxp3 expression via demethylation of a CpG island in Foxp3. J. Cell. Physiol. © 2014 Wiley Periodicals, Inc.

Description: Primary cilia are sensory organelles that provide a feedback mechanism to restrict Wnt signaling in the absence of endogenous Wnt activators. Abnormal Wnt signaling has been shown to result in polycystic kidney disease (PKD) although the exact mechanism has been debated. Previously, we reported that the calcium channel CaV1.2 functions in primary cilia. In this study, we show that CaV1.2 expression level is regulated by Wnt signaling. This occurs through modulation of mitochondrial mass and activity resulting in increased reactive oxygen species which generate oxidative DNA lesions. We found that the subsequent cellular DNA damage response triggers increased CaV1.2 expression. In the absence of primary cilia where Wnt signaling is upregulated, we found that CaV1.2 is overexpressed as a compensatory mechanism. We show for the first time that CaV1.2 knockdown in zebrafish results in classic primary cilia defects including renal cyst formation, hydrocephalus, and left-right asymmetry defects. Our study shows that suppressed Wnt signaling prevents CaV1.2 expression ultimately resulting in PKD phenotypes. Thus, CaV1.2 expression is tightly regulated through Wnt signaling and plays an essential sensory role in primary cilia necessary for cellular homeostasis. J. Cell. Physiol. 9999: XX–XX, 2014. © 2014 Wiley Periodicals, Inc.

Description: To reduce environmental impact of cement production, granulated blast furnace slag (GBFS) can be used as a cement clinker substitution. Its reactivity of can be increased by adequate fine grinding. The impact of different conventional laboratory-scaled mills (ball mill, vertical mill, and roller press) on the material properties was researched. Granulometrical, structural, and calorimetrical investigations were done. It could be demonstrated that at the same fineness the grinding unit does not significantly influence the cement performance like hydration heat release and compressive strength. On the other hand, particle properties like particle size distribution, particle shape, and thus workability characteristics of composite cements have been changed. The replacement of ordinary produced cement by ground granulated blast furnace slag (GGBFS) in the cement production reduces negative impacts on the environment. By adequate fine grinding the reactivity of GGBFS as a cement linker substitution could be enhanced. The influence of grinding devices on the reactivity of GGBFS is investigated and evaluated.

Description: The impact of polydispersity index (PDI) values and doses of two sodium polyacrylates on grinding limits and particle size distributions of highly concentrated limestone suspensions was studied. The results indicated that viscosity and agglomerate particle size, and thus the apparent grinding limit, depended on the sodium polyacrylate dose as well as PDI value of sodium polyacrylate. The higher the sodium polyacrylate dose, the lower the viscosity and the smaller the apparent grinding limit were. The smallest primary particle size, the smallest apparent grinding limit, the narrowest agglomerate particle size distribution, and the lowest viscosity curve were obtained with sodium polyacrylate having a low PDI. Ultrafine grinding of limestone in stirred media mills is an attractive method for nano-CaCO 3 production. Effects of polydispersity index and doses of two sodium polyacrylates on the grinding limits of limestone suspensions were studied. Sodium polyacrylate with a low polydispersity index turned out to be an effective grinding aid for limestone.

Description: A method was sought that allows for coordinating the particle grinding time and the particle size in a closed milling circuit. This can be achieved by placing the input of the return flow from an end classifier at an intermediate position of the tube mill length. A simple model to estimate the efficiency of the approach is proposed. It is shown that such an optimal position exists and depends on the classifier efficiency. One of the key problems in improving the efficiency of grinding is how to match the particle size and the necessary grinding time. A model for optimizing the position of the circulating load input in a closed milling circuit with a tube mill is proposed. It is shown that such an optimum does exist and can yield a considerable gain in the circuit capacity.

Description: Phenomena related to sieving of non-spherical particles are investigated numerically in two batch apparatuses and on a horizontally aligned continuous sieve by particle-based simulation approaches in the framework of the discrete element method. The feed material is approximated by complex-shaped particles composed of clustered spheres. Comparisons are made with regard to the passage through the screen as well as the segregation and transportation on the screen. Results for passage are compared to data from literature, where simulations with spherical particles were performed of a laboratory-scale sieve operated with non-spherical quarry rock particles. Additionally, variations in screen inclination are investigated. Experimental results are matched by the simulations. A distinctive influence of particle shape on flow rates and residence times is identified. Despite many modeling advances, phenomenological models still lack the ability to represent dynamic processes within the unit operation sieving. Here, discrete element method simulations of complex-shaped particles on two batch apparatuses and a continuously operated sieve are performed. Phenomena related to sieving of non-spherical particles are investigated numerically.

Description: The process and end-use behavior of ground material not only depends on the particle size or specific surface but is also influenced by the particle shape. Particle shaping can be done in a stirred media mill if the stress intensity is not enough to break the particle, so mainly abrasion and chipping are carried out. Low-stress intensity milling is presented in a dry stirred media mill to reveal the particle base rounding and surface roughness reduction. The main parameters of the product-related stress model are connected to the descriptive parameters of particle shaping. Batch grinding of limestone particles with narrow size distribution was carried out with different milling parameters. The product-related stress model can be used for the description of particle shaping. The rounded shape and smooth surface of particles is necessary for many processes. A stirred media mill was used in dry mode to produce rounded or smooth surface particles in a size range of a few hundred microns without significant size reduction. An evaluation method was created to show the effect of the milling parameters on the particle shaping in relation to the product-related stress model.

Description: The influence of several process parameters like milling time, ball-to-beaker volume ratio, diameter of milling balls, and rotation frequency on the Knoevenagel condensation of vanillin and barbituric acid in planetary ball mills was investigated. These parameters determine the amount of energy provided for the reaction. Additionally, numerical simulations were carried out to describe the stress conditions in detail and to compute the drive power and energy transfer which cannot be measured directly. The mill and experimental parameters were modeled by the discrete element method with adequate coefficients of friction and restitution required to describe the powder behavior in the system. The coefficients were determined by correlation of experiments and simulations. Mechanochemical synthesis may offer green solutions to organic synthesis. Using the Knoevenagel reaction under solvent-free conditions in a planetary ball mill, the results are correlated with discrete element method simulations to prove the dependency of the reaction yield on stress conditions. Smaller balls were more time and energy efficient due to higher power values achieved by high stress frequencies.

Description: The aim of this work is to clarify the complex interactions and various influencing parameters regarding the material behavior of deep-frozen bulk solids. Additionally, a better evaluation of the material behavior in practical applications needs to be ensured. As material samples, hard coal, iron ore, and sand were selected, and cube-shaped samples saturated with water were deep-frozen for defined periods of time. Subsequently, the influences of the residence time in the freezer and of the particle size on the material behavior were analyzed by carrying out uniaxial compression tests. An economically and technically efficient method is needed to overcome the problems associated with frozen coal. Hard coal, iron ore, and sand samples saturated with water were deep-frozen for defined periods of time. Uniaxial compression tests were carried out to analyze the influence of the particle size and the residence time in the freezer on the material behavior.

Description: A general overview for the process development and manufacturing of engineered drug particles by size reduction techniques is provided. Drug compound and drug product characteristics are reviewed, which may need an engineered drug particle size. Drug particle attributes are introduced which are affected by size reduction techniques and potentially require additional downstream processing effort. The development of drug particle size specifications is described, including adequate size control. A brief overview is given on dry- and wet-milling technologies currently applied in drug development and manufacturing, including selected application examples. Finally, the processing of highly active drug compounds is introduced with respect to equipment design, worker protection, cleaning and waste management to minimize worker exposure and environmental impact. A brief overview is given on drug compound and drug product characteristics impacted by drug particle size. Size reduction techniques applied after chemical synthesis and isolation processing are introduced for the engineering of drug particle size. The repercussions for the processing of highly active drug compounds are overviewed with regard to worker exposure and environmental impact.

Description: The production of nanosuspensions via stirred-media milling has proved to be an effective method to overcome bioavailability challenges of poorly water-soluble active pharmaceutical ingredients (APIs). The fine-grinding of the APIs cinnarizine and fenofibrate was investigated. Important process parameters were varied and the influence on product quality was studied. Different challenges were identified prior to and during milling: Foaming inside the milling equipment and agglomeration of fenofibrate occurred, and a long-term grinding experiment for cinnarizine revealed an increase in particle size, caused by ripening of the drug crystals during milling. Thus, the stability against ripening during the process is a newly identified challenge for the fine-grinding of APIs. The influence of process parameters on product quality for the milling of the active pharmaceutical ingredients fenofibrate and cinnarizine was investigated. Besides the well-known challenge regarding stabilization of the newly formed (nano)particles, a further challenge occurred: the ripening of the cinnarizine crystals during or shortly after the milling process.

Description: The interest in finer, dry products has led to the development of more efficient jet milling processes. The first part of the paper describes the thermodynamic basic principles for generation and application of steam, and compressed gases. Practical experience has shown that in the aspired fineness range stabilization during grinding provides an enormous potential for improvement. Some selected examples demonstrate that the energy requirement can be reduced by a factor of more than two by suitable choice of stabilizers. Application of single-stage, uncooled low-pressure compressors for jet milling offers an economical alternative to double-stage, cooled compressors for fine grinding. Stabilization during grinding provides an enormous potential for improvement. Selected examples demonstrate that the energy requirement can be reduced by a factor of 〉 2 with suitable stabilizers.

Description: Wet grinding processes, especially in terms of nanoparticle production, are extremely energy intensive, and for hard and abrasive product materials, high grinding media wear is produced. In order to minimize energy consumption, the choice of the process parameters is crucial. The decision on the process parameters often depends on experience or a certain number of laboratory- or pilot-scale experiments. Here, an enhanced stress energy model is used, which enables the prediction of optimum process parameters for inorganic materials in wet stirred media milling based on results for a different grinding material. The use of this model to select the process parameters is also valid with regard to grinding media wear: The grinding media wear reaches minimum values if the optimum parameters pertaining to the minimum specific energy are chosen based on this enhanced stress energy model, especially if soft products are ground. An enhanced stress energy model is used for the prediction of optimum process parameters for inorganic materials in wet stirred media milling based on results for a different grinding material. Optimization of the process parameters leads to minimum wear values for a certain product fineness, especially for materials that are softer than the grinding media material.