Interactomic analysis of REST/NRSF and implications of its functional links with the transcription suppressor TRIM28 during neuronal differentiation

RE-1 silencing transcription factor (REST) is a transcriptional repressor that regulates gene expression by binding to repressor element 1. However, despite its critical function in physiology, little is known about its interaction proteins. Here we identified 204 REST-interacting proteins using affinity purification and mass spectrometry. The interactome included proteins associated with mRNA processing/splicing, chromatin organization, and transcription. The interactions of these REST-interacting proteins, which included TRIM28, were confirmed by co-immunoprecipitation and immunocytochemistry, respectively. Gene Ontology (GO) analysis revealed that neuronal differentiation-related GO terms were enriched among target genes that were co-regulated by REST and TRIM28, while the level of CTNND2 was increased by the knockdown of REST and TRIM28. Consistently, the level of CTNND2 increased while those of REST and TRIM28 decreased during neuronal differentiation in the primary neurons, suggesting that CTNND2 expression may be coregulated by both. Furthermore, neurite outgrowth was increased by depletion of REST or TRIM28, implying that reduction of both REST and TRIM28 could promote neuronal differentiation via induction of CTNND2 expression. In conclusion, our study of REST reveals novel interacting proteins which could be a valuable resource for investigating unidentified functions of REST and also suggested functional links between REST and TRIM28 during neuronal development.1121Ysciescopu

Effects of Rosiglitazone on the Expression of PPAR-&#947 and the Production of IL-6 and IL-8 in Acute Lung Injury Model Using Human Pulmonary Epithelial Cells

Purpose: Peroxisome proliferator-activated receptor (PPAR)-γ ligand is known to repress the expression of pro-inflammatory mediators. However, it is unclear how it affects PPAR-γ expression and the inflammatory response in the human lung. We investigated the effects of rosiglitazone (synthetic PPAR-γ ligand) on the PPAR-γ expression and on the IL-6 and IL-8 production in acute lung injury model using human lung epithelial cells.Methods: A549 and Beas-2B cells were pre-treated with rosiglitazone and/or BADGE (selective PPAR-γ antagonist) and then treated with media control or cytokine mixture including TNF-α, IL-1 β, and IFN-γ. PPAR-γ expression was analyzed in cell lysates by Western blot. IL-6 and IL-8 production was measured in the culture supernatants by ELISA.Results: PPAR-γ expression was identified in all experimental groups except for the control. The cytokine mixture-induced IL-6 and IL-8 production was significantly inhibited by pre-treatment with rosiglitazone (P<0.01). However, this inhibitory effect of rosiglitazone was not reversed by BADGE.Conclusion: These suggest that rosiglitazone induces the PPAR-γ expression and it may inhibit the cytokine mixture-induced IL-6 and IL-8 production through the PPAR-γ independent pathway. The inhibitory mechanisms of rosiglitazone on the cytokine mixture-induced IL-6 and IL-8 production in human alveolar and bronchial epithelial cells remain to be further investigated.Keywords: Rosiglitazone, PPAR-γ, IL-6, IL-8, Acute lung injur

Effects of Rosiglitazone on the Expression of PPAR-&#947 and on the Production of IL-6 and IL-8 in Acute Lung Injury Model Using Human Pulmonary Epithelial Cells

Purpose: Peroxisome proliferator-activated receptor (PPAR)-γ ligand is known to repress the expression of pro-inflammatory mediators. However, it is unclear how it affects PPAR-γ expression and the inflammatory response in the human lung. We investigated the effects of rosiglitazone (synthetic PPAR-γ ligand) on the PPAR-γ expression and on the IL-6 and IL-8 production in acute lung injury model using human lung epithelial cells.Methods: A549 and Beas-2B cells were pre-treated with rosiglitazone and/or BADGE (selective PPAR-γ antagonist) and then treated with media control or cytokine mixture including TNF-α, IL-1β, and IFN-γ. PPAR-γ expression was analyzed in cell lysates by Western blot. IL-6 and IL-8 production was measured in the culture supernatants by ELISA.Results: PPAR-γ expression was identified in all experimental groups except for the control. The cytokine mixture-induced IL-6 and IL-8 production was significantly inhibited by pre-treatment with rosiglitazone (P<0.01). However, this inhibitory effect of rosiglitazone was not reversed by BADGE. Conclusion: These suggest that rosiglitazone induces the PPAR-γ expression and it may inhibit the cytokine mixture-induced IL-6 and IL-8 production through the PPAR-γ independent pathway. The inhibitory mechanisms of rosiglitazone on the cytokine mixture-induced IL-6 and IL-8 production in human alveolar, and bronchial epithelial cells remain to be further investigated.Keywords: Rosiglitazone, PPAR-γ expression, IL-6, IL-8, Acute lung injur

Coefficient of standardized total tract digestibility of phosphorus in oilseed meals and distillers dried grains in growing-finishing pigs

This study was conducted to determine the coefficient of total tract standardized digestibility (CTTSD) of phosphorus (P) in oilseed meals and distillers dried grains (DDG) fed to growing-finishing pigs. Twelve barrows (initial bodyweight (BW) standard deviation, 52.25 +/- 2.57 kg) were allocated individually to metabolism cages. The experimental design was a 12 x 8 incomplete Latin square with 12 dietary treatments and eight replication periods. The diets were formulated individually with dehulled soybean meal produced in Korea (SBM-KD), soybean meal produced in India (SBM-I), soybean meal produced in Korea (SBM-K), corn high-protein distiller dried grains (HPDDGs), tapioca distillers dried grains (TDDG), canola meal (CAM), corn germ meal (CGM), copra meal (CM), palm kernel meal (PKM), sesame meal (SM), perilla meal (PM), and a P-free diet. Intake of P was highest in SM and PM. Excretion of P was reduced in y ascending order as HPDDG, TDDG and CGM; SBM-K; and SM and PM. The CTTAD of P was higher in CGM than SBM-K, TDDG, SM and PM. HPDDG and CGM showed greater CTTSD of P than SBM-K, CAM and PM. Digestible concentration of P on CTTSD (CTTSD-P) of P was greater in PM and CAM than the others except for SBM-KD. In summary, PM could be utilized as an alternative feedstuff to SBM, but its usage is regarded only as a source of P. In addition, the results of the current study would provide valuable information for formulating pig feed with precise P utilization in ingredients using mixed diet

A Preliminary Risk Assessment on the Development of a Small-Scaled Floating Power Plant

This paper introduces a preliminary risk assessment method carried out on a newly developed floating power plant. The small-scale floating power plant has been developed to provide electric power for areas on demand, and this is a kind of a new concept system which is not clearly classified in the maritime industry. To grant the feasibility for this novel system design, a set of risk assessment activities is essentially required, and in this context, a hazard identification (HAZID) study is conducted at the very early stage of the plant design. The aims of this HAZID study are to verify the inherent safety of the initial plant design and to provide any recommendations on the next design stages. For this purpose, the potential hazards are identified in view of personnel, structural, and asset effects in association with the operation of the power plant and all the identified hazards and relevant risks are assessed with the defined criteria using a simple risk matrix. As a result, the risk or safety level of the conceptual plant design is estimated, and some design changes are suggested to give a better balance between the safety and the cost of the plant system. Overall, this paper shows how the primitive risk assessment techniques are utilized as a practical engineering tool in the development of the marine system

TonEBP/NFAT5 stimulates transcription of HSP70 in response to hypertonicity

While hyperosmolality of the kidney medulla is essential for urinary concentration, it imposes a great deal of stress. Cells in the renal medulla adapt to the stress of hypertonicity (hyperosmotic salt) by accumulating organic osmolytes. Tonicity-responsive enhancer (TonE) binding protein (TonEBP) (or NFAT5) stimulates transcription of transporters and a synthetic enzyme for the cellular accumulation of organic osmolytes. We found that dominant-negative TonEBP reduced expression of HSP70 as well as the transporters and enzyme. Near the major histocompatibility complex class III locus, there are three HSP70 genes named HSP70-1, HSP70-2, and HSC70t. While HSP70-1 and HSP70-2 were heat inducible, only HSP70-2 was induced by hypertonicity. In the 5' flanking region of the HSP70-2 gene, there are three sites for TonEBP binding. In cells transfected with a reporter plasmid containing this region, expression of luciferase was markedly stimulated in response to hypertonicity. Coexpression of the dominant-negative TonEBP reduced the luciferase expression. Mutating all three sites in the reporter plasmid led to a complete loss of induction by hypertonicity. Thus, TonEBP rather than heat shock factor stimulates transcription of the HSP70-2 gene in response to hypertonicity. We conclude that TonEBP is a master regulator of the renal medulla for cellular protection against high osmolality via organic osmolytes and molecular chaperones.open12

Disrupted-in-schizophrenia 1 (DISC1) and Syntaphilin collaborate to modulate axonal mitochondrial anchoring

In neuronal axons, the ratio of motile-to-stationary mitochondria is tightly regulated by neuronal activation, thereby meeting the need for local calcium buffering and maintaining the ATP supply. However, the molecular players and detailed regulatory mechanisms behind neuronal mitochondrial movement are not completely understood. Here, we found that neuronal activation-induced mitochondrial anchoring is regulated by Disrupted-in-schizophrenia 1 (DISC1), which is accomplished by functional association with Syntaphilin (SNPH). DISC1 deficiency resulted in reduced axonal mitochondrial movement, which was partially reversed by concomitant SNPH depletion. In addition, a SNPH deletion mutant lacking the sequence for interaction with DISC1 exhibited an enhanced mitochondrial anchoring effect than wild-type SNPH. Moreover, upon neuronal activation, mitochondrial movement was preserved by DISC1 overexpression, not showing immobilized response of mitochondria. Taken together, we propose that DISC1 in association with SNPH is a component of a modulatory complex that determines mitochondrial anchoring in response to neuronal activation.117Ysciescopu

Physicochemical, antioxidant, and sensorial properties of peach snacks prepared from different cultivars

High potentiality of medicinal benefits of peach have increased its demand, however, supply of fresh fruit to meet the demand is challenging as it grows in specific climatic regions and in particular season. Preparation of varieties of processed peach products could be a good option to supply in lean season. Objective of the study was to assess the quality characteristics of peach snacks prepared from 11 different cultivars. We investigated the physicochemical (soluble solid, titratable acidity, hardness and dry yield), antioxidant (DPPH radical scavenging capacity and total phenolics content) and sensorial properties (color, flavor, texture, sweetness and overall acceptance) of peach snacks prepared from 11 peach cultivars. Peach snacks of different cultivars evaluated in this study showed substantial variations in antioxidant capacity,  physicochemical and sensorial properties. Some of the samples showed higher physicochemical properties while the others contained better antioxidant capacity or sensory properties. Results of this study reveal that quality peach snacks having different properties could be prepared by drying thin slices of fresh fruits of different cultivars. However, supplementary studies on cost effective techniques of peach snacks preparation and variation in nutritive and medicinal properties of processed products could increase the application of the findings of this experiment.Key words: Antioxidant, peach snack, physicochemical, sensory property

Toxicity assessment of modified Cry1Ac1 proteins and genetically modified insect-resistant Agb0101 rice

Insect-resistant Agb0101 rice was recently developed by modifying the cry1ac1 gene by changing codon usage changes relative to the native truncated cry1ac1 gene. To assess the toxicity of genetically modified Agb0101 rice, we conducted bioinfomational comparisons of the amino acid sequences that are not similar to known toxic proteins. Sufficient quantities of mCry1Ac1 protein were produced in Escherichia coli for in vitro evaluation and animal study. We compared the amino acid sequences and molecular mass. There have the same amino acid sequences and molecular masses after purifying the modified Cry1Ac1 (mCry1Ac1) protein from highly expressed bacteria and genetically modified rice were identical. We also investigated the acute and 90-days oral toxicities. No adverse effects were observed in mice following acute oral exposure to 2,000 mg/ kg body weight mCry1Ac1 protein of body weight and 90 days oral exposure to Agb0101. These results indicate that mCry1Ac1 proteins and Agb0101 rice demonstrate no adverse effects in these tests when applied via gavage and feed, respectively.Key words: Modified Cry1Ac1, food safety assessment, toxicity, insect- resistant rice Agb0101