Ucma, a direct transcriptional target of Runx2 and Osterix, promotes osteoblast differentiation and nodule formation

SummaryObjectiveRunt-related transcription factor 2 (Runx2) and Osterix (Osx) are the master transcription factors in bone formation. Nonetheless, genes acting downstream of both Runx2 and Osx have yet to be fully characterized. Here, we investigate the downstream targets of both Runx2 and Osx in osteoblasts.Materials and methodsDNA microarray analysis was conducted on calvarial RNA from wild-type, Runx2 heterozygous, Osx heterozygous, and Runx2/Osx double heterozygous embryos. Expression and transcriptional responses of the selected target gene were analyzed in MC3T3-E1 osteoblastic cells.ResultsThe expression of unique cartilage matrix-associated protein (Ucma) was decreased in Runx2/Osx double heterozygous embryos. In contrast, Ucma expression was increased in osteoblasts overexpressing both Runx2 and Osx. Ucma expression was initiated mid-way through osteoblast differentiation and continued throughout the differentiation process. Transcriptional activity of the Ucma promoter was increased upon transfection of the cells with both Runx2 and Osx. Runx2-and Osx-mediated activation of the Ucma promoter was directly regulated by Runx2-and/or Sp1-binding sites within its promoter. During osteoblast differentiation, the formation of mineralized nodules in Ucma-overexpressing stable clones occurred earlier and was more enhanced than that in the mock-transfected control. Mineralized nodule formation was strongly augmented in the cells cultured in a medium containing secretory Ucma proteins.ConclusionUcma is a novel downstream gene regulated by both Runx2 and Osx and it stimulates osteoblast differentiation and nodule formation

Ethyl Alcohol Extract of Hizikia fusiforme Induces Caspase-dependent Apoptosis in Human Leukemia U937 Cells by Generation of Reactive Oxygen Species

Purpose: Hizikia fusiforme is renowned for the possession of anti-inflammatory and anti-oxidant properties. In this study, the role of the ethyl alcohol extract of H. fusiforme (EAHF) in the induction of apoptosis in human leukemia U937 cells was investigated.Methods: Protein expression was investigated by Western blot analysis. Cell viability and apoptosis were analyzed by an MTT assay and flow cytometric analysis. Caspase activity was analyzed using a caspase-specific kit.Results: EAHF suppressed the proliferation of U937 cells in a dose-dependent manner. This effect was closely related to the induction of apoptosis via the downregulation of IAP family members such as IAP-1, IAP-2 and XIAP, as well as Bcl-2 proteins. The results also showed that caspases play an essential role in EAHF-induced apoptosis by generating of reactive oxygen species (ROS). In addition, ROS scavenging by N-acetyl-L-cysteine (NAC) and glutathione (GSH) decreased EAHF-induced apoptosis via the suppression of caspase activity. Although EAHF induced the phosphorylation of mitogenactivated protein kinases (MAPKs), treatment with MAPK inhibitors did not affect EAHF-induced apoptosis.Conclusion: These results suggest that EAHF induces apoptosis in U937 cells via ROS-dependent caspase activation.Keywords: Hizikia fusiforme, Apoptosis, Caspase, Reactive oxygen specie

Rheological, chemical and DSC thermal characteristics of different types of palm oil/palm stearin-based shortenings

This study was carried out to evaluate the physical and chemical properties of different types of shortenings, formulated by mixing refined, bleached, and deodorized palm oil and palm stearin (PO:PS) in the following ratios: 100:0, 80:20, 60:40, 50:50, 40:60 and 20:80 and 0:100. The properties of experimental and commercial shortenings were investigated using four different analytical techniques, namely high performance liquid chromatography (HPLC), gas chromatography (GC), differential scanning calorimetry (DSC) and controlled stress rheometer. In addition, iodine value (IV) analysis was carried out. The results revealed that the prominent fatty acids in the products were palmitic (44.88-61.91%), oleic (26.24-39.14%) and linoleic (6.13-11.68%). At the same time, triacyglycerols (TAG), such as OOO, OOP and OOS, were found to decrease, while PPO increased due to the increase in the palm stearin content of the shortenings. Higher viscosity and more storage (G′) or loss (G″) modulus properties were noted in the experimental and commercial shortenings containing higher and lower concentrations of palm stearin and palm oil, respectively. Certain parameters such as the onset, peak and endset temperatures (ºC) were detected for both the melting and cooling data. However, increasing the palm stearin concentrations in the samples was shown to have caused increases in the endset temperature and peak height, and vice versa. Thus, chemical and physical properties of the formulated shortenings may influence the quality of baked products

Specific detection of Pythium aphanidermatum from hydroponic nutrient solution by booster PCR with DNA primers developed from mitochondrial DNA

Pythium aphanidermatum causes damping-off and root rot of vegetable crops in hydroponic systems. A DNA probe was isolated and modified from a library of HindIII-digested mitochondrial DNA of P aphanidermatum that strongly hybridized to DNA of R aphanidermatum and weakly hybridized to DNA of Pythium deliense. Cross-hybridizing sequences were absent from DNA of plants and other related fungi. The probe detected as little as 5 ng of P. aphanidermatum DNA and 250 ng of P deliense DNA in slot-blot assays. P. aphanidermatum was detected by a hybridization assay of total DNA extracted directly from infected roots. A pair of oligonucleotide primers P1 and RP2, which allowed amplification of a specific 0.65 kb DNA fragment of P. aphanidernatum using polymerase chain reaction (PCR), was designed from a specific DNA probe. Specific amplification of this fragment from P. aphanidermatum was highly sensitive, detecting template DNA as low as 0.1 pg total DNA by booster PCR. Specific booster PCR amplification using P1 and RP2 was successful in detecting P. aphanidermatum in naturally infected nutrient solution and roots of vegetables in a field hydroponic system

The Aharonov-Bohm effect for massless Dirac fermions and the spectral flow of Dirac type operators with classical boundary conditions

We compute, in topological terms, the spectral flow of an arbitrary family of self-adjoint Dirac type operators with classical (local) boundary conditions on a compact Riemannian manifold with boundary under the assumption that the initial and terminal operators of the family are conjugate by a bundle automorphism. This result is used to study conditions for the existence of nonzero spectral flow of a family of self-adjoint Dirac type operators with local boundary conditions in a two-dimensional domain with nontrivial topology. Possible physical realizations of nonzero spectral flow are discussed.Comment: 15 pages, 6 figures. Submitted to Theoretical and Mathematical Physics. v2: A change has been made to the paragraph describing the previous work of M. Prokhorov

Na+/K+-ATPase α-subunit ( nkaα) isoforms and their mRNA expression levels, overall nkaαα protein abundance, and kinetic properties of Nka in the skeletal muscle and three electric organs of the electric eel, electrophorus electricus

10.1371/journal.pone.0118352PLoS ONE103e011835

A cross validation of Consumer-Based Brand Equity (CBBE) with Private Labels in Spain

Molinillo,S., Ekinci, Y., Japutra, A. (2014)'A cross validation of Consumer-Based Brand Equity (CBBE) with Private Labels in Spain'. in Martínez-López, Gázquez-Abad, J.C. and Sethuraman, R. J.A. (eds.) Advances in National Brand and Private Label Marketing. Second International Conference, 2015. Springer Proceedings in Business and Economics, pp. 113-125In recent years a number of Consumer-Based Brand Equity (CBBE) models and measurement scales have been introduced in the branding literature. However, examinations of brand equity in Private Labels (PL) are rather limited. This study aims to compare the validity of the two prominent CBBE models those introduced by Yoo and Donthu (2001) and Nam et al. (2011). In order to test the models and make this comparison, the study collected data from 236 respondents who rated private labels in Spain. A list of 30 different fashion and sportswear PL was introduced to respondents. These brands do not make any reference to the retail store in which they are sold. Research findings suggest that the extended CBBE model introduced by Nam et al. (2011) and Ciftci et al. (2014) is more reliable and valid than Yoo and Donthu’s model for assessing PL. Theoretical contributions and managerial implications are discussed.Universidad de Málaga. Campus de Excelencia Internacional Andalucía Tech

Incipient Separation in Shock Wave Boundary Layer Interactions as Induced by Sharp Fin

The incipient separation induced by the shock wave turbulent boundary layer interaction at the sharp fin is the subject of present study. Existing theories for the prediction of incipient separation, such as those put forward by McCabe (1966) and Dou and Deng (1992), can have thus far only predicting the direction of surface streamline and tend to over-predict the incipient separation condition based on the Stanbrook's criterion. In this paper, the incipient separation is firstly predicted with Dou and Deng (1992)'s theory and then compared with Lu and Settles (1990)' experimental data. The physical mechanism of the incipient separation as induced by the shock wave/turbulent boundary layer interactions at sharp fin is explained via the surface flow pattern analysis. Furthermore, the reason for the observed discrepancy between the predicted and experimental incipient separation conditions is clarified. It is found that when the wall limiting streamlines behind the shock wave becomes\ aligning with one ray from the virtual origin as the strength of shock wave increases, the incipient separation line is formed at which the wall limiting streamline becomes perpendicular to the local pressure gradient. The formation of this incipient separation line is the beginning of the separation process. The effects of Reynolds number and the Mach number on incipient separation are also discussed. Finally, a correlation for the correction of the incipient separation angle as predicted by the theory is also given.Comment: 34 pages; 9 figure

Pontin functions as an essential coactivator for Oct4-dependent lincRNA expression in mouse embryonic stem cells

The actions of transcription factors, chromatin modifiers and noncoding RNAs are crucial for the programming of cell states. Although the importance of various epigenetic machineries for controlling pluripotency of embryonic stem (ES) cells has been previously studied, how chromatin modifiers cooperate with specific transcription factors still remains largely elusive. Here, we find that Pontin chromatin remodelling factor plays an essential role as a coactivator for Oct4 for maintenance of pluripotency in mouse ES cells. Genome-wide analyses reveal that Pontin and Oct4 share a substantial set of target genes involved in ES cell maintenance. Intriguingly, we find that the Oct4-dependent coactivator function of Pontin extends to the transcription of large intergenic noncoding RNAs (lincRNAs) and in particular linc1253, a lineage programme repressing lincRNA, is a Pontin-dependent Oct4 target lincRNA. Together, our findings demonstrate that the Oct4-Pontin module plays critical roles in the regulation of genes involved in ES cell fate determination.111311Ysciescopu