Preparation and properties of OSA-modified taro starches and their application for stabilizing Pickering emulsions.

In this study, the octenylsuccinylated taro starches (OSTS) with different degree of substitution (DS, from 0.009 to 0.032) were prepared and their structural properties such as granule size, wettability and morphology were studied. The purpose of this work was to elucidate the OSTS with different DS using as particle stabilizers for Pickering emulsions, and the effect of DS on the stability, droplet size, microstructure and rheological properties of OSTS-stabilized emulsions were investigated. Octenylsuccinic anhydride (OSA) modification had slight effects on the morphology or granule size of taro starch, but markedly increased the contact angle from 25.4° to 70.1°. Octenylsuccinylation significantly improved the emulsifying capacity of taro starch granules, and thus OSTS-stabilized emulsions formed at higher DS exhibited better stability. Droplet size distribution results and microscopic observations revealed that OSTS-emulsion prepared at DS of 0.032 had the smallest droplet size and most uniform distribution compared with the other emulsions. The rheological results indicated that both OSTS-emulsions (DS, from 0.009 to 0.032) showed shear-thinning behavior as a non-Newtonian fluid, and the viscosities of emulsions were progressively improved with the increase of DS. Moreover, the G' and G″ values of OSTS-emulsions increased with increasing DS, reflecting the enhanced viscoelastic properties and exhibiting an improved rigidity of the emulsions. The above results suggested that higher-DS favored the formation of superior OSTS-emulsions, and thus OSTS with a high DS (DS ≥ 0.018) can be used for preparing stable Pickering emulsions.

Production of vinegar from purple sweet potato in a liquid fermentation process and investigation of its antioxidant activity.

Acetobacter pasteurianus JST-S was screened from solid fermented grains of vinegar in China, identified by molecular analysis, and used for the production of purple sweet potato vinegar using purple sweet potato as the substrate. By orthogonal experiment, maximum total acid concentration (4.26% [v/v]) was achieved under optimized conditions as follows: fermentation time, 3.5 days; ethanol content, 9% v/v; and inoculum size, 8% v/v. During the production of purple potato vinegar, the anthocyanin concentration decreased from 652.07 to 301.73 µg/mL. The antioxidant activity of products, including diphenyl-picryl hydrazide radical-scavenging capacity (above 60%), reducing power (above 0.47), and hydroxyl radical-scavenging capacity (above 46%), showed positive linear regression (P < 0.01), which could be related with the changes in anthocyanin concentration and antioxidant activities at different stages of vinegar fermentation. The acetic acid and other non-phenolic antioxidants in purple sweet potato vinegar may have contributed to the antioxidant activities. Results of these studies may provide a reference for the industrial production of vinegar by liquid fermentation of purple sweet potato.

Scalable Production of Mechanically Robust Antireflection Film for Omnidirectional Enhanced Flexible Thin Film Solar Cells.

Antireflection (AR) at the interface between the air and incident window material is paramount to boost the performance of photovoltaic devices. 3D nanostructures have attracted tremendous interest to reduce reflection, while the structure is vulnerable to the harsh outdoor environment. Thus the AR film with improved mechanical property is desirable in an industrial application. Herein, a scalable production of flexible AR films is proposed with microsized structures by roll-to-roll imprinting process, which possesses hydrophobic property and much improved robustness. The AR films can be potentially used for a wide range of photovoltaic devices whether based on rigid or flexible substrates. As a demonstration, the AR films are integrated with commercial Si-based triple-junction thin film solar cells. The AR film works as an effective tool to control the light travel path and utilize the light inward more efficiently by exciting hybrid optical modes, which results in a broadband and omnidirectional enhanced performance.

Cytotoxicity, tumor targeting and PET imaging of sub-5 nm KGdF4 multifunctional rare earth nanoparticles.

Ultrasmall sub-5 nm KGdF4 rare earth nanoparticles were synthesized as multifunctional probes for fluorescent, magnetic, and radionuclide imaging. The cytotoxicity of these nanoparticles in human glioblastoma U87MG and human non-small cell lung carcinoma H1299 cells was evaluated, and their application for in vitro and in vivo tumor targeted imaging has also been demonstrated.

Long-term-stable near-infrared polymer dots with ultrasmall size and narrow-band emission for imaging tumor vasculature in vivo.

Fluorescent nanoprobes have become one of the most promising classes of materials for cancer imaging. However, there remain many unresolved issues with respect to the understanding of their long-term colloidal stability and photostability in both biological systems and the environment. In this study, we report long-term-stable near-infrared (NIR) polymer dots for in vivo tumor vasculature imaging. NIR-emitting polymer dots were prepared by encapsulating an NIR dye, silicon 2,3-naphthalocyanine bis(trihexylsilyloxide) (NIR775), into a matrix of polymer dots, poly[2-methoxy-5-(2-ethylhexyloxy)-1,4-phenylenevinylene] (MEH-PPV), using a nanoscale precipitation method. The prepared NIR polymer dots were sub-5 nm in diameter, exhibited narrow-band NIR emission at 778 nm with a full width at half-maximum of 20 nm, and displayed a large Stokes shift (>300 nm) between the excitation and emission maxima. In addition, no significant uptake of the prepared NIR polymer dots by either human glioblastoma U87MG cells or human non-small cell lung carcinoma H1299 cells was detected. Moreover, these NIR polymer dots showed long-term colloidal stability and photostability in water at 4 °C for at least 9 months, and were able to image vasculature of xenografted U87MG tumors in living mice after intravenous injection. These results thus open new opportunities for the development of whole-body imaging of mice based on NIR polymer dots as fluorescent nanoprobes.

Physicochemical properties, antioxidant activities and protective effect against acute ethanol-induced hepatic injury in mice of foxtail millet (Setaria italica) bran oil.

This study was designed to investigate physicochemical characterization of the oil extracted from foxtail millet bran (FMBO), and the antioxidant and hepatoprotective effects against acute ethanol-induced hepatic injury in mice. GC-MS analysis revealed that unsaturated fatty acids (UFAs) account for 83.76% of the total fatty acids; in particular, the linoleic acid (C18:2) is the predominant polyunsaturated fatty acid (PUFA), and the compounds of squalene and six phytosterols (or phytostanols) were identified in unsaponifiable matter of FMBO. The antioxidant activity examination of FMBO in vitro showed highly ferric-reducing antioxidant power and scavenging effects against DPPH· and HO· radicals. Furthermore, the protective effect of FMBO against acute hepatic injuries induced by ethanol was verified in mice. In this, intragastric administration with different dosages of FMBO in mice ahead of acute ethanol administration could observably antagonize the ethanol-induced increases in serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), triglyceride (TG), and the hepatic malondialdehyde (MDA) levels, respectively, along with enhanced hepatic superoxide dismutase (SOD) levels relative to the control. Hepatic histological changes were also observed and confirmed that FMBO is capable of attenuating ethanol-induced hepatic injury.

GRIM-19 opposes reprogramming of glioblastoma cell metabolism via HIF1α destabilization.

The metabolism that sustains cancer cells is adapted preferentially to glycolysis, even under aerobic conditions (Warburg effect). This effect was one of the first alterations in cancer cells recognized as conferring a survival advantage. In this study, we show that gene associated with retinoid-interferon-induced mortality-19 (GRIM-19), which was previously identified as a tumor suppressor protein associated with growth inhibition and cell apoptosis, contributes to the switch between oxidative and glycolytic pathways. In parallel to this, vascular endothelial growth factor, which promotes neovascularization, is also regulated. We have identified hypoxia-inducible factor 1α (HIF1α) as the downstream factor of GRIM-19 in human glioblastoma cell lines. Downregulation of GRIM-19 promotes HIF1α synthesis in a STAT3-dependent manner, which acts as a potential competitive inhibitor for von Hippel-Lindau (pVHL)-HIF1α interaction, and thereby prevents HIF1α from pVHL-mediated ubiquitination and proteasomal degradation. Taken together, it is concluded that GRIM-19, a potential tumor suppressor gene, performs its function in part via regulating glioblastoma metabolic reprogramming through STAT3-HIF1α signaling axis, and this has added new perspective to its role in tumorigenesis, thus providing potential strategies for tumor metabolic therapy.

Function of GRIM-19, a mitochondrial respiratory chain complex I protein, in innate immunity.

Mitochondria respiratory chain (RC), consisting of five multisubunit complexes, is crucial for cellular energy production, reactive oxygen species generation, and regulation of apoptosis. Recently, a few mitochondrial proteins have been reported to be essential for innate immunity, but the function of mitochondrial RC in innate immunity is largely unknown. By knock-out of GRIM-19, a newly identified subunit protein of mitochondrial complex I, in mice, we found that heterogeneous mice (GRIM-19(+/-)) are prone to spontaneous urinary tract infection, mostly by Staphylococcus saprophyticus. Macrophages derived from these mice have compromised mitochondrial complex I activity and increased reactive oxygen species level. Bacterial infection induces a rapid up-regulation of GRIM-19 and complex I activity in the wild-type macrophages, but both are reduced in the macrophages from GRIM-19(+/-) mice. These cells also have decreased intracellular killing ability against S. saprophyticus. The defects for this probably occur in the fusion of bacteria to lysosome, but not in the bacterial engulfment and macrophage migration. In addition, production of proinflammatory cytokines, such as interleukin (IL)-1, IL-12, IL-6, and interferon (IFN)-γ, induced by both bacterial infection and lipopolysaccharide (LPS) and monodansylcadaverine treatment, is also decreased in the GRIM19(+/-) macrophages. Inhibition of mitochondrial RC activity by inhibitors shows a similar reduction on the cytokine production. Due to low cytokine production, the inflammatory response caused by in vivo bacterial challenge in the bladders of GRIM-19(+/-) mice is compromised. This study provides genetic evidence for a critical role of mitochondrial RC in innate immunity.

Depletion of GRIM-19 accelerates hepatocellular carcinoma invasion via inducing EMT and loss of contact inhibition.

UNLABELLED: Genes associated with retinoid-interferon-induced mortality 19 (GRIM-19) was identified as a tumor suppressor protein associated with apoptosis and growth inhibition. Here, we report that the expression levels of GRIM-19 are significantly attenuated in hepatocellular carcinoma (HCC) patients with deteriorating differentiation states, hepatic capsule invasion and microvascular invasion, suggesting the potential role of GRIM-19 not only at the origin but also in the invasive progression of HCCs. To dissect the possible mechanisms by which GRIM-19 regulates tumor cell invasion, we established the hepatic HL-7702 and HCC Huh-7 cell lines stably depleted of GRIM-19. Results show that downregulation of GRIM-19 induces a morphological transformation resembling epithelial-mesenchymal transition (EMT) as well as aberrant expression of epithelial and mesenchymal molecular markers. Additionally, these cells lose contact inhibition, a phenomenon of cessation of cell migration in contact with neighboring cells, as assessed by cell imaging, growth curve and S-phase transition in confluent conditions. CONCLUSION: Our observations demonstrate a novel mechanistic insight into a critical role of GRIM-19 in HCC invasive potential.

Identification of alternatively spliced GRIM-19 mRNA in kidney cancer tissues.

Gene associated with Retinoid-Interferon-induced Mortality (GRIM)-19 was originally identified as a regulatory gene necessary for interferon-beta and retinoic acid-induced cell death. Further studies revealed that GRIM-19 is a subunit of mitochondrial respiratory chain complex I. Previous studies show that the expression of GRIM-19 is lost or severely depressed in a number of primary renal cell carcinomas (RCCs) and in some urinogenital tumors. Four point mutations were found in the GRIM-19 gene in mitochondria-rich (Hürthle) tumors and one point mutation was reported in RCC. In this study, we report an alternatively splicing form of GRIM-19 mRNA with intron 3 by reverse transcriptase PCR. This splicing variant is found in kidney tumor tissues but not in matched normal tissues. Furthermore, we found that in addition to GRIM-19, the protein level of NDUFS3, which is another mitochondrial complex I subunit, was also diminished in kidney tumor tissues when compared with paired normal tissues. Our finding suggested that the alternative splicing form of GRIM-19 is tumor tissue specific.

SCG10-like protein (SCLIP) is a STAT3-interacting protein involved in maintaining epithelial morphology in MCF-7 breast cancer cells.

STAT (signal transducer and activator of transcription) 3 is a key contributor to cancer cell migration and invasion, with excessive STAT3 activity promoting growth arrest, cell-cell dissociation and increased migration of breast cancer epithelial cells. The STAT3-regulated mechanisms involved in this process, however, are not fully defined. Previously, we had revealed SCLIP [SCG10 (superior cervical ganglia protein 10)-like protein] as a novel STAT3-interacting protein. In the present study, we show that STAT3 binds the C-terminal tubulin-associating region of SCLIP. In a search for a function of SCLIP, we show that SCLIP was down-regulated during OSM (oncostatin M) treatment in MCF-7 cells, which also stimulates epithelial morphology loss. SCLIP knockdown likewise triggered a loss of epithelial morphology which included reduced E-cadherin expression. We found that STAT3 was required to maintain SCLIP stability. Furthermore, inhibition of OSM-induced STAT3 activity preserved SCLIP expression and MCF-7 epithelial monolayers. Taken together, we propose that a STAT3-SCLIP interaction is required to preserve SCLIP stability and contributes to the maintenance of normal epithelial morphology. Disruption of the STAT3-SCLIP interaction with OSM may contribute to cytokine-mediated loss in cell-cell attachment and morphology transition in MCF-7 cells.

Stat3 promotes directional cell migration by regulating Rac1 activity via its activator betaPIX.

Stat3 is a member of the signal transducer and activator of transcription family, which is important for cytokine signaling as well as for a number of cellular processes including cell proliferation, anti-apoptosis and immune responses. In recent years, evidence has emerged suggesting that Stat3 also participates in cell invasion and motility. However, how Stat3 regulates these processes remains poorly understood. Here, we find that loss of Stat3 expression in mouse embryonic fibroblasts leads to an elevation of Rac1 activity, which promotes a random mode of migration by reducing directional persistence and formation of actin stress fibers. Through rescue experiments, we demonstrate that Stat3 can regulate the activation of Rac1 to mediate persistent directional migration and that this function is not dependent on Stat3 transcriptional activity. We find that Stat3 binds to betaPIX, a Rac1 activator, and that this interaction could represent a mechanism by which cytoplasmic Stat3 regulates Rac1 activity to modulate the organization of actin cytoskeleton and directional migration.

Mitochondria and calcium signaling in embryonic development.

Calcium (Ca2+) is a simple but critical signal for controlling various cellular processes and is especially important in fertilization and embryonic development. The dynamic change of cellular Ca2+ concentration and homeostasis are tightly regulated. Cellular Ca2+ increases by way of Ca2+ influx from extracellular medium and Ca2+ release from cellular stores of the endoplasmic reticulum (ER) and sarcoplasmic reticulum (SR). The elevated Ca2+ is subsequently sequestered by expelling it out of the cell or by pumping back to the ER/SR. Mitochondria function as a power house for energy production via oxidative phosphorylation in most eukaryotes. In addition to this well-known function, mitochondria are also recognized to regulate Ca2+ homeostasis through different mechanisms. Although critical roles of Ca2+ signaling in fertilization and embryonic development are known, the involvement of mitochondria in these processes are not fully understood. This review is focused on the role of mitochondrial respiratory chain complex I in the regulation of Ca2+ signaling pathway and gene expression in embryonic development, especially on the new findings in the cardiac development of Xenopus embryos. The data demonstrate that mitochondria modulate Ca2+ signaling and the Ca2+-dependent NFAT pathway and its target gene which are essential for embryonic heart development.

Cytokine profiling and Stat3 phosphorylation in epithelial-mesenchymal interactions between keloid keratinocytes and fibroblasts.

We previously reported an increase in signal transducer and activator of transcription 3 (Stat3) activation in keloid fibroblasts, which contributes to collagen production, cell proliferation, and migration. We further investigated the effect of epithelial-mesenchymal interaction on Stat3 in normal and keloid fibroblasts in noncoculture and coculture conditions. pY705 Stat3 was higher in keloid fibroblasts compared to normal fibroblasts in noncoculture. However, a more drastic decrease in pY705 Stat3 was observed in keloid fibroblasts compared to normal fibroblasts when cocultured with their respective keratinocytes over 5 days. To explore this paracrine effect, we examined the secretion of cytokines by cytokine arrays. Altered cytokine production was detected in keloid fibroblasts and keratinocytes, either in noncoculture or coculture conditions. IL-6, IL-8, monocyte chemoattractant protein-1, tissue inhibitor of metalloproteinases (TIMPs)-1, and TIMP-2 were major cytokines detected. Angiogenin, oncostatin M (OSM), vascular endothelial cell growth factor, IGF-binding protein-1, osteoprotegerin, and transforming growth factor-beta2 were present in keloid keratinocyte-fibroblast coculture, but absent in normal keratinocyte-fibroblast coculture. Only IL-6 and OSM stimulated strong pY705 Stat3 and cell proliferation in both normal and keloid fibroblasts. Other cytokines increased proliferation of keloid fibroblasts, but not normal fibroblasts, suggesting an altered state in keloid fibroblasts. Multiple cytokines likely contribute to keloid pathogenesis and a combinatorial neutralizing antibody/cytokine therapy may be effective in ameliorating keloid scars.

FoxO1 inhibits leptin regulation of pro-opiomelanocortin promoter activity by blocking STAT3 interaction with specificity protein 1.

Leptin controls food intake and energy expenditure by regulating hypothalamic neuron activities. Leptin exerts its actions through complex signaling pathways including STAT3 phosphorylation, nuclear translocation, and binding to target gene promoter/cofactor complexes. Deficient or defective leptin signaling leads to obesity, which may be caused by insufficient leptin levels and/or resistance to leptin signaling. To understand the molecular mechanisms of leptin resistance, we studied the regulation of pro-opiomelanocortin (POMC) gene expression by leptin. We show that phospho-STAT3 activates POMC promoter in response to leptin signaling through a mechanism that requires an SP1-binding site in the POMC promoter. Furthermore, FoxO1 binds to STAT3 and prevents STAT3 from interacting with the SP1.POMC promoter complex, and consequently, inhibits STAT3-mediated leptin action. Our study suggests that leptin action could be inhibited at a step downstream of STAT3 phosphorylation and nuclear translocation, and provides a potential mechanism of leptin resistance in which an increased FoxO1 antagonizes STAT3-mediated leptin signaling.

Oct4 is expressed in human gliomas and promotes colony formation in glioma cells.

There is increasing evidence that self-renewal capacity of cancer cells is critical for carcinogenesis; hence, it is vital to examine the expression and involvement of self-renewal regulatory genes in these cells. Here, we reported that Oct4, a well-known regulator of self-renewal in embryonic stem cells, was highly expressed in human gliomas and glioma cell lines, and the expression levels were increased in parallel with increasing glioma grades. In in vitro cell cultures, Oct4 was only expressed in rat C6 glioma cells and rat neural stem cells but not in rat brain differentiated cells. Downregulation of Oct4 expression by RNA interference in C6 cells was associated with reduced cell proliferation and colony formation. Further analysis revealed that Oct4 could upregulate phosphorylation of Stat3 to promote tumor cell proliferation. Overexpression of Oct4 in C6 cells increased the expression of nestin but decreased the expression of GFAP suggesting that Oct4 might inhibit the differentiation of glioma cells. Our findings may provide further evidence for the stem cell theory of carcinogenesis. In contrast, the results might also imply that Oct4 contributes to the existence of undifferentiated cells in gliomas.

Nuclear import of Pin1 is mediated by a novel sequence in the PPIase domain.

Pin1 actively regulates diverse biological/pathological processes, but little is known about the regulatory mechanisms of its cellular localization. In this study, we report that the endogenous Pin1 is distributed in both nucleus and cytoplasm. We found that point mutations of several basic amino acids in the PPIase domain of Pin1 significantly compromise its nuclear localization. Such inhibition is independent of Pin1 enzymatic activity, and is mainly due to the defects in the nuclear import. A novel sequence harboring these residues was identified as a putative nuclear localization signal (NLS) of Pin1. Importin alpha5 of the nuclear import machinery was found to interact with Pin1.

NFAT directly regulates Nkx2-5 transcription during cardiac cell differentiation.

BACKGROUND INFORMATION: The transcription factor NFAT (nuclear factor of activated T-cell) family comprises important regulators in immuno-responses and mouse embryonic development, including early cardiovascular and heart valve development. The mechanism involved, however, is not fully understood. Nkx2-5 (NK2 transcription factor related, locus 5) is one of the earliest genes expressed in early cardiac progenitor cells and is essential for heart tube development by control of a subset of cardiac muscle-specific genes. Previously we found that downregulation of mitochondrial respiratory chain complex I caused severe cardiac deficiencies during heart tube development in Xenopus embryos associated with compromised Nkx2-5 expression. However, the heart defects and Nkx2-5 expression could be rescued by a constitutively activated NFAT, suggesting a possible link between NFAT and Nkx2-5 during early heart development. RESULTS: In the present study, we demonstrate that NFAT regulates Nkx2-5 expression in both mouse ES (embryonic stem) cells and P19 cells, a mouse model for embryonic differentiation. We found that there are six core NFAT-binding elements in the 5' regulatory region of the Nkx2-5 gene. Although NFAT is able to bind directly to all but one of these elements, it activates Nkx2-5 transcription only via a specific binding site in the distal enhancer region. Interestingly, the transcriptional activity of NFAT is largely dependent on the co-factor GATA (GATA-binding transcription factor), which binds to an element adjacent to this key NFAT-binding site. Furthermore, binding of the endogenous NFAT to this particular site was observed during cardiac differentiation in mouse ES and P19 cells. CONCLUSIONS: The results suggest that Nkx2-5 is a direct target of NFAT that co-ordinates with other transcription factors such as GATA4 to regulate Nkx2-5 during cardiogenesis.

NF-kappaB p52, RelB and c-Rel are transported into the nucleus via a subset of importin alpha molecules.

In resting cells NF-kappaB transcription factors are retained in the cytoplasm as latent inactive complexes, until they are activated and rapidly transported into the nucleus. We show that all NF-kappaB proteins are imported into the nucleus via a subset of importin alpha isoforms. Our data indicate that the NF-kappaB components of the classical and alternative pathways have somewhat different specifities to importin alpha molecules. Based on the results from binding experiments of in vitro-translated and Sendai virus infection-induced or TNF-alpha-stimulated endogenous NF-kappaB proteins, it can be predicted that the specifity of NF-kappaB proteins to importin alpha molecules is different and changes upon the composition of the imported dimer. p52 protein binds directly to importin alpha3, alpha4, alpha5 and alpha6 and c-Rel binds to importin alpha5, alpha6 and alpha7 via a previously described monopartite nuclear localization signals (NLSs). Here we show that RelB, instead, has a bipartite arginine/lysine-rich NLS that mediates the binding of RelB to importin alpha5 and alpha6 and subsequent nuclear translocation of the protein. Moreover, we show that the nuclear import of p52/RelB heterodimers is mediated exclusively by the NLS of RelB. In addition, we found that the NLS of p52 mediates the nuclear import of p52/p65 heterodimers.

GRIM-19 is essential for maintenance of mitochondrial membrane potential.

GRIM-19 was found to copurify with complex I of mitochondrial respiratory chain and subsequently was demonstrated to be involved in complex I assembly and activity. To further understand its function in complex I, we dissected its functional domains by generating a number of deletion, truncation, and point mutants. The mitochondrial localization sequences were located at the N-terminus. Strikingly, deletion of residues 70-80, 90-100, or the whole C-terminal region (70-144) led to a loss of mitochondrial transmembrane potential (DeltaPsim). However, similar deletions of another two complex I subunits, NDUFA9 and NDUFS3, did not show such effect. We also found that deletion of the last 10 residues affected GRIM-19's ability to be assembled to complex I. We constructed a dominant-negative mutant containing the N-terminal 60 and the last C-terminal 10 residues, which could be assembled into complex I, but failed to maintain normal DeltaPsim. Cells overexpressing this mutant did not spontaneously undergo cell death, but were sensitized to apoptosis induced by cell death agents. Our results demonstrate that GRIM-19 is required for electron transfer activity of complex I, and disruption of DeltaPsim by GRIM-19 mutants enhances the cells' sensitivity to apoptotic stimuli.