Search

Canadian double-muscled Large White pigs are characterized by notable muscle mass, showing high daily gain and lean rate and good meat quality. In order to identify the major genes or proteins involved in muscle hyperplasia and hypertrophy, three pairs of full-sib pigs with extreme muscle mass difference from Canadian Large White were selected as experimental animals at 3 months age. The phenotypic differences of longissimus dorsi muscles (LD) were investigated with microarray and proteomics (2-DE, MALDI-TOF-MS), and results were verified by real-time PCR and western bolting respectively. The gene expressing profiling identified 57 and 260 and 147 differently expressed genes (DEGs) from the three pairs respectively with Bayesian statistics and significant analysis of microarrays (SAM) (p<0.05, q<0.05, fold>2). From the network of these DEGs, some major genes were displayed, such as EGF, PPARG, FN1, SERPINE1, MYC, JUN, involved in Wnt, MAPK and TGF-β signal pathway respectively, which mainly participated in cell differentiation and proliferation. In parallel, proteomics analyses revealed 50 differently expressed protein (DEP) spots with mass spectrum, and 33 spots of them were found annotated, which took part in energy metabolism and the structure and contraction of muscle fiber. In brief, our integrated study provides a good foundation for the further study on the genetic mechanism of the double muscle traits in pigs., (Copyright © 2015 Elsevier B.V. All rights reserved.)

Carbohydrates are known to be involved in a wide range of biological and pathological processes. However, due to the presence of multiple hydroxyl groups, carbohydrate recognition is a particular challenge. Herein, we reported an ultrasensitive solid-phase microextraction (SPME) probe based on phenylboronic acid (PBA) functionalized carbon nanotubes (CNTs) for direct in vitro or in vivo recognition of carbohydrates in biofluids as well as semi-solid biotissues. The coating of the proposed probe possessed a 3D interconnected porous architecture formed by the stacking of CNTs. As a result, the binding capacity toward carbohydrates was excellent. The proposed approach was demonstrated to be much superior to most carbohydrate sensors, including higher sensitivity, wider linear range, and excellent qualitative ability in multi-carbohydrate systems. Thus, this approach opens up new avenues for the facile and efficient recognition of carbohydrates for important applications such as glycomics.

A combination of nitrogen-doped ordered mesoporous polymer (NOMP) and stainless steel wires led to highly sensitive, selective, and stable solid phase microextraction (SPME) fibers by in situ polymerization for the first time. The ordered structure of synthesized NOMP coating was illustrated by transmission electron microscopy (TEM) and X-ray diffraction (XRD), and microscopy analysis by scanning electron microscopy (SEM) confirmed a homogenous morphology of the NOMP-coated fiber. The NOMP-coated fiber was further applied for the extraction of organochlorine pesticides (OCPs) with direct-immersion solid-phase microextraction (DI-SPME) method followed by gas chromatography-mass spectrometry (GC-MS) quantification. Under the optimized conditions, low detection limits (0.023-0.77 ng L(-1)), a wide linear range (9-1500 ng L(-1)), good repeatability (3.5-8.1%, n=6) and excellent reproducibility (1.5-8.3%, n=3) were achieved. Moreover, the practical feasibility of the proposed method was evaluated by determining OCPs in environmental water samples with satisfactory recoveries., (Copyright © 2015 Elsevier B.V. All rights reserved.)

Microporous organic polymers (MOPs) have emerged as a new class of functional porous materials with unique characteristics and potential uses in diverse areas. However, the field of MOPs for gas chromatographic (GC) separations has not been well explored. Herein, a MOP namely KAPs-1 was dynamic coated onto a capillary column for the first time. The fabricated column exhibited a nonpolar nature and the column efficiency for n-dodecane was up to 7769 plates m(-1). The KAPs-1 coated column showed high GC separation performance for a series of volatile organic compounds (VOCs) including the challenging ethylbenzene and xylene isomers, which could not be resolved at baseline on the commercial 5% phenyl polysiloxane stationary phase. Moreover, the relative standard deviations for five replicate determinations of the studied analytes were 0.0-0.6%, 0.9-3.2%, 1.1-5.9%, 0.8-3.7% for retention time, peak area, peak height and peak width, respectively. To investigate the interaction between some analytes and the stationary phase, thermodynamic and kinetic parameters were also evaluated. The results of this study show it is very promising to utilize MOPs as stationary phases for capillary GC., (Copyright © 2015 Elsevier B.V. All rights reserved.)

This study presents the preparation and characterization of a nanoscale Davankov-type hyper-crosslinked-polymer (HCP) as an adsorbent of benzene-ring-containing dyes and organic pollutants. HCP nanoparticles post-crosslinked from a poly(DVB-co-VBC) precursor were synthesized in this study, possessing ultrahigh surface area, hydrophobicity and stability. The as-synthesized Davankov-type HCP exhibited a rapid and selective adsorption ability towards the benzene-ring-containing dyes due to its highly conjugated structure. Besides, for the first time, the prepared HCP nanoparticles were adopted for the adsorption of nonpolar organic pollutants by means of solid-phase microextraction (SPME). Owing to its high hydrophobicity, diverse pore size distribution and highly conjugated structure, a 10 μm HCP coating exhibited excellent adsorption abilities towards benzene-ring-containing polycyclic aromatic hydrocarbons (PAHs) and benzene series compounds (benzene, toluene, ethylbenzene and o-xylene; abbreviated to BTEX) and to highly hydrophobic long-chain n-alkanes. Finally, the HCP-nanoparticles-coated SPME fiber was applied to the simultaneous analysis of five PAHs in environmental water samples and satisfactory recoveries were achieved. The findings could provide a new benchmark for the exploitation of superb HCPs as effective adsorbents for SPME or other adsorption applications.

Development of facile and effective methods for fabrication of high-performance solid phase microextraction (SPME) fibres remains a great challenge. Herein, a new class of ordered mesoporous polymers (OMPs) in situ coated on a stainless steel wire were successfully developed and utilized as a highly sensitive and stable SPME fibre for the first time. Because of the highly ordered mesoporous structure of its OMP coating, the π-π interactions and the dispersion forces, the OMP-coated SPME fibre exhibited much better extraction properties as compared to the commercial PDMS fibre. The findings could provide a new benchmark for preparing well-defined porous materials for the SPME application.

Here we report the successful utilization of the stepwise ligand exchange strategy for the improvement of adsorption ability of a series of bio-MOFs. The fast extraction rate and the different adsorption performances of the three bio-MOF coatings were dominated by their pore structures.

Mesoporous TiO2 nanoparticles were synthesized with the hydrothermal method and characterized by powder X-ray diffraction (PXRD) and transmission electron microscope (TEM). Then a superior solid-phase microextraction (SPME) fiber was fabricated by sequentially coating the stainless steel fiber with silicone sealant film and mesoporous TiO2 powder. The developed fiber possessed a homogeneous surface and a long life-span up to 100 times at direct immersing (DI) extraction mode. Under the optimized conditions, the extraction efficiencies of the self-made 17 μm TiO2 fiber for six organochlorine pesticides (OCPs) were higher than those of the two commercial fibers (65 μm PDMS/DVB and 85 μm PA fibers) which were much thicker than the former. As for analytical performance, low detection limits (0.08-0.60 ng L(-1)) and wide linearity (5-5000 ng L(-1)) were achieved under the optimal conditions. The repeatabilities (n=5) for single fiber were between 2.8 and 12.3%, while the reproducibilities (n=3) of fiber-to-fiber were in the range of 3.7-15.7%. The proposed fiber was successfully applied to the sensitive analysis of OCPs in real water samples and four of the six analytes were detected from the rainwater and the lake water samples., (Copyright © 2015 Elsevier B.V. All rights reserved.)

Metal-organic frameworks (MOFs) have received great attention as novel sorbents due to their fascinating structures and intriguing potential applications in various fields. In this work, a MIL-101(Cr)-coated solid-phase microextraction (SPME) fiber was fabricated by a simple direct coating method and applied to the determination of volatile compounds (BTEX, benzene, toluene, ethylbenzene, m-xylene and o-xylene) and semi-volatile compounds (PAHs, polycyclic aromatic hydrocarbons) from water samples. The extraction and desorption conditions of headspace SPME (HS-SPME) were optimized. Under the optimized conditions, the established methods exhibited excellent extraction performance. Good precision (<7.7%) and low detection limits (0.32-1.7 ng L(-1) and 0.12-2.1 ng L(-1) for BTEX and PAHs, respectively) were achieved. In addition, the MIL-101(Cr)-coated fiber possessed good thermal stability, and the fiber can be reused over 150 times. The fiber was successfully applied to the analysis of BTEX and PAHs in river water by coupling with gas chromatography-mass spectrometry (GC-MS). The analytes at low concentrations (1.7 and 10 ng L(-1)) were detected, and the recoveries obtained with the spiked river water samples were in the range of 80.0-113% and 84.8-106% for BTEX and PAHs, respectively, which demonstrated the applicability of the self-made fiber., (Copyright © 2014 Elsevier B.V. All rights reserved.)

The aim of the present study was to determine whether tumor necrosis factor (TNF)-α regulates the expression levels of excitatory amino-acid transporters (EAATs) in primary astrocytes and its roles in brain ischemia. Exogenous TNF-α was administered to pure cultured astrocytes and the expression levels of EAAT1, EAAT2 and glial fibrillary acidic protein (GFAP) were evaluated. The results showed that TNF-α at 10 ng/ml enhanced the expression of EAAT2 in a time-dependent manner, while the expression levels of EAAT1 and GFAP did not change. To determine whether the elevation in the levels of the EAAT2 protein induced by TNF-α had a beneficial effect on ischemic insult, TNF-α was applied to in vitro models of cerebral ischemia; the treatment was observed to increase neuronal viability. The present results suggest that a relatively short-term application of an optimal concentration of TNF-α may protect neurons against ischemic injury by elevating the expression of EAAT2 in astrocytes.

We study localized surface waves at the interface between linear dielectric and biased centrosymmetric photorefractive (CP) crystals. If the propagation constant b is fixed, the energy of localized surface waves increases with the order of localized surface waves. For low b values, the considerable part of the energy of localized surface waves is concentrated in the linear dielectric and decreases with an increase in b. For high b values, the part of the energy of localized surface waves concentrated in the nonlinear CP crystals is always higher than that in the linear dielectric and increases with b. The stability properties of these localized surface waves are also discussed in detail.

Oncoprotein p28GANK knockdown by RNA interference (RNAi) can induce hepatoma cells apoptosis. However, the mechanisms have not been well defined yet. In the present study the p28GANK knockdown-induced apoptosis in HepG2 cells was prevented by caspase-9 inhibitor (Z-LEHD-FMK). During the knockdown of p28GANK, mitochondrial translocation of Bax, loss of mitochondrial transmembrane potential (DeltaPsim) and release of cytochrome c were observed. In this study, the activation of p38 was found to be critical for the p28GANK knockdown-induced apoptosis, as suggested by the finding that pharmacological inhibition of p38 with SB203580 suppressed the redistribution of Bax, the loss of DeltaPsim and the apoptosis. Moreover, generation of reactive oxygen species (ROS) contributed to the cell death because N-acetyl-L-cystenine (NAC), a ROS scavenger, suppressed the phosphorylation of p38 and the apoptosis. Our studies established the signaling pathway of p28GANK knockdown-induced apoptosis in HepG2 cells, namely, mitochondrial dysfunction mediated by p38 downstream of intracellular ROS generation.

Transfection of DNA vaccines with chemokines may recruit dendritic cells (DCs) locally to capture the antigenic genes and their gene products to generate enhanced CD8(+) cytotoxic T lymphocytes (CTLs). In this study, we investigated the effects of macrophage inflammatory protein (MIP)-1 alpha, MIP-3 alpha, and MIP-3beta on human immunodeficiency virus (HIV) Gag DNA vaccination. The chemokine plasmids markedly enhanced the local infiltration of inflammatory cells and increased the presence of CD11c(+) B7.2(+)-activated DCs. MIP-1 alpha and MIP-3 alpha were potent adjuvants in augmenting CTLs and afforded strong protection to immunized animals against challenge with vaccinia virus expressing Gag (vv-Gag). However, decreased humoral response was observed. MIP-3beta plasmid did not dramatically alter immunity. The chemokine inoculation time with respect to DNA vaccine priming was also investigated. The injection of pMIP-3 alpha three days before Gag plasmid (pGag) vaccination markedly increased specific CTLs compared with simultaneous injection and led to higher protection against vv-Gag. Immunity was also shifted toward a T-helper type-1 (Th1) response. In contrast, inoculation with pMIP-3 alpha three days after pGag vaccination shifted immunity toward a Th2 response. Our data suggest that administration of a chemokine with DNA vaccines offers a valuable strategy to modulate the efficacy and polarization of specific immunity and that chemokine-antigen timing is critical in determining overall biological effects.

Transfection of DNA vaccines with chemokines may recruit dendritic cells (DCs) locally to capture the antigenic genes and their gene products to generate enhanced CD8 + cytotoxic T lymphocytes (CTLs). In this study, we investigated the effects of macrophage inflammatory protein (MIP)-1α, MIP-3α, and MIP-3β on human immunodeficiency virus (HIV) Gag DNA vaccination. The chemokine plasmids markedly enhanced the local infiltration of inflammatory cells and increased the presence of CD11c + B7.2 + -activated DCs. MIP-1α and MIP-3α were potent adjuvants in augmenting CTLs and afforded strong protection to immunized animals against challenge with vaccinia virus expressing Gag (vv-Gag). However, decreased humoral response was observed. MIP-3β plasmid did not dramatically alter immunity. The chemokine inoculation time with respect to DNA vaccine priming was also investigated. The injection of pMIP-3α three days before Gag plasmid (pGag) vaccination markedly increased specific CTLs compared with simultaneous injection and led to higher protection against vv-Gag. Immunity was also shifted toward a T-helper type-1 (Th1) response. In contrast, inoculation with pMIP-3α three days after pGag vaccination shifted immunity toward a Th2 response. Our data suggest that administration of a chemokine with DNA vaccines offers a valuable strategy to modulate the efficacy and polarization of specific immunity and that chemokine-antigen timing is critical in determining overall biological effects., (Copyright © 2007 The American Society of Gene Therapy. Published by Elsevier Inc. All rights reserved.)

Controlled release of granulocyte-macrophage colony-stimulating factor (GM-CSF) protein by albumin-heparin microparticles administered via intramuscular vaccination in conjunction with HIV DNA vaccines stimulated HIV Gag-specific immune responses. In the murine model, Gag-specific cytotoxic T lymphocyte (CTL) and T helper (Th) responses were significantly enhanced by administration of murine GM-CSF microparticles. This effect was comparable to a GM-CSF encoded plasmid. In three of four rhesus monkeys, enhancement of Gag-specific antibody (Ab), Th, and CTL responses was observed 1 month after the first immunization with coadministration of human GM-CSF microparticles and HIV Gag plasmid. The second, third, and fourth booster immunizations, however, did not increase the Gag-specific immune responses. Subsequent application of Gag protein in complete Freund's adjuvant (CFA) significantly enhanced Ab and Th, but not CTL. However, Gag-specific CTL response was triggered by cytokine and Gag p55-encapsulated microparticles in all animals. The strategy of priming immune responses by coadministration of cytokine microparticles and DNA vaccines, followed by boosting with cytokine and antigen protein-encapsulated microparticles, may prove effective in improving an HIV DNA vaccine design.

Signal regulatory protein alpha (SIRPalpha) is an Ig superfamily protein whose cytoplasmic region contains immunoreceptor tyrosine-based inhibitory motif (ITIM), which when tyrosine phosphorylated binds the SH2-domain containing phosphatase 2 (SHP-2). Both SIRPalpha and SHP-2 are highly expressed in brain. Murine cerebellar cells cultured on SIRPalpha-coated surface exhibit enhanced neurite outgrowth and SIRPalpha is localized at sites of synaptogenesis in postnatal mouse brain. In this study, we show that nerve growth factor (NGF) stimulation resulted in elevated SIRPalpha expression during PC12 differentiation. We also show that NGF-induced morphological differentiation, but not growth arrest response, was inhibited by ectopic SIRPalpha expression. PC12 cells stably expressing SIRPalpha proliferated more rapidly than mock-transfected cells. The activity of c-jun N-terminal kinase (JNK) decreased in SIRPalpha-transfected PC12 cells, whereas nuclear factor-kappaB (NF-kappaB) activity increased. Collectively, our results suggest that SIRPalpha may stabilize synaptic connections by inhibiting improper neurite outgrowth and might realize its neuronal function, at least in part, by modulating JNK and NF-kappaB activity.

Purpose: To investigate the sensitivity, specificity, and spatial distribution of the product of p28 gene (p28(GANK) protein) in human hepatocellular carcinoma (HCC) and nonhepatocellular carcinomas in relation to immunostaining with Cytokeratin 18 (CK18), alpha-fetoprotein (AFP), and Hepatocyte paraffin 1 (HepPar1)., Method: In this retrospective study, formalin-fixed paraffin-embedded tissues from 24 HCCs, five intrahepatic cholangiocarcinomas (ICC), five combined hepatocellular cholangiocarcinomas (C-HCC-CC) and mine metastatic hepatic carcinomas (MHC) were immunostained for p28(GANK) as well as CK18, AFP and HepPar1. Only cases with more intensified staining in carcinoma contrast to the adjacent liver tissues were accepted as positive., Result: In HCC, p28(GANK) was expressed restrictively in hepatocytes of both para-lesion and carcinoma liver tissues, while absent in the bile duct epithelial cells, Kupffer cells, and other interstitial cells. The positive staining of p28(GANK) was noted in 16 (66.7%) specimens of HCC and three (60.0%) specimens of C-HCC-CC, and no specific lesion staining was found in all tested specimens of ICC and MHC. Sensitivity and specificity for hepatocyte-originated carcinoma were, respectively, 65.5% and 100% for p28(GANK), 79.3% and 85.2% for CK18, 20.7% and 100% for AFP, 79.3% and 92.0% for HepPar1., Conclusion: The hepatocytic staining for p28(GANK) is clearly useful in differentiating hepatocyte-originated carcinoma from non-HCC. p28(GANK) may be used as an ancillary marker for the diagnosis of HCC.

Small-diameter microporous polyurethane vascular prostheses with an average pore size of between 5 and 30mum at the outer surfaces and 30mum at the luminal surface were prepared. Thirty-two PU and 8 expanded polytetrafluoroethylene (ePTFE) prostheses were implanted into the abdominal aorta of rats for periods ranging from 1 to 8 weeks. Harvested prostheses were analysed histologically and morphologically. The progress of endothelial-like cells and the extent of infiltration of perigraft tissues were quantified. All of the prostheses showed fast growth of endothelial-like cells in the second week, with the PU prostheses having an external pore size of 30mum producing the highest rate. It was also during the second week that perigraft tissue grew most significantly into the prosthetic structure. This coincident may suggest the importance of rapid tissue regeneration for the early endothelial healing. The role of the ingrowth perigraft tissues is likely to support and stabilize the neointima. The thickening of neointima was mainly located at the vicinity of the proximal anastomoses of some of the PU prostheses and was unrelated with the extent of perigraft tissue infiltration. In the PU prostheses, a complete lining of endothelial-like cells was achieved by the end of 4 weeks. Expanded PTFE prostheses displayed smooth, thin intima, very limited tissue ingrowth, and incomplete coverage of endothelial-like cells.