CXCL14 is a chemokine that has previously been implicated in insulin resistance in mice. In humans, the role of CXCL14 in metabolic processes is not well established, and we sought to determine whether CXCL14 is a risk susceptibility gene important in fetal programming of metabolic disease. For this purpose, we investigated whether CXCL14 is differentially regulated in human umbilical cords of infants with varying birth weights. We found an elevated expression of CXCL14 in human low birth weight (LBW) cords, as well as in cords from nutritionally restricted Macaca fascicularis macaques. To further analyze the regulatory mechanisms underlying the expression of CXCL14, we examined CXCL14 in umbilical cord-derived mesenchymal stem cells (MSCs) that provide an in vitro cell-based system amenable to experimental manipulation. Using both whole frozen cords and MSCs, we determined that site-specific CpG methylation in the CXCL14 promoter is associated with altered expression, and that changes in methylation are evident in LBW infant-derived umbilical cords that may indicate future metabolic compromise through CXCL14.
An iodinated urethane polymer that does not require addition of X-ray attenuating additives to impart X-ray opacity was synthesized and characterized for biomedical applications. A new X-ray opaque diiodo- compound, namely, 2,2-(2,5-diiodobenzene-1,4-diyl)bis(oxy)]diethanol (DBD), was synthesized by iodinating hydroquinone bis(2-hydroxyethyl) ether and this compound was used as chain extender during polyurethane synthesis so that X-ray opacity could be imparted to the polymer formed. X-ray opaque polyurethane (XPU) was synthesized by reacting 1,6-diisocyanatohexane with poly(hexamethylene carbonate) diol and DBD. X-ray opacity of XPU was measured with a fluoroscopy machine using BaSO4 filled polyurethane as controls. Radiographic images showed that XPU sample had X-ray opacity equivalent to 15wt% BaSO4 filled polymer. In vivo imaging in a rabbit model showed that the material could be readily distinguishable from bones. XPU was found to be hemocompatible and non-cytotoxic to L929 fibroblast cell lines. Optical transparency measurements using UV -Vis spectrophotometer showed that XPU transmitted 85% of visible light.
We describe a scalable method for producing continuous graphene networks by tape casting surfactant-stabilized aqueous suspensions of functionalized graphene sheets. Similar to all other highly connected graphene-containing networks, the degree of overlap between the sheets controls the tapes electrical and mechanical properties. However, unlike other graphene-containing networks, the specific surface area of the cast tapes remains high (>400 m(2)·g(-1)). Exhibiting apparent densities between 0.15 and 0.51 g·cm(-3), with electrical conductivities up to 24 kS·m(-1) and tensile strengths over 10 MPa, these tapes exhibit the best combination of properties with respect to density heretofore observed for carbon-based papers, membranes, or films.
Vascular grafts are devices intended to replace compromised arteries in the body and grafts made of polyethylene terephthalate (PET) fabric have been used mainly for synthetic grafting procedures involving medium to large diameter vascular grafts. Though porosity of the graft permits tissue in-growth, it would lead to bleeding through the graft walls immediately after implantation. So it is essential to seal the pores either by preclotting with patients own blood or by other sealing materials prior to implantation in order to prevent blood leakage through the graft wall. Biodegradable hydrogel materials are ideal candidates for this purpose. Apart from sealing the pores, they offer biocompatible and low-thrombogenic surfaces when coated on vascular graft. In the present study, a biodegradable hydrogel, derived from oxidized alginate and gelatin, has been deposited on PET grafts by dip coating and were characterized for its efficacy on sealing the pores of the graft. Water permeability in the static and pulsatile conditions, burst strength, in vitro cell culture cytotoxicity, hemocompatibility, and endothelial cell adhesion and proliferation of the coated grafts were investigated. Results showed that the alginate dialdehyde cross-linked gelatin hydrogel was nontoxic, hemocompatible, and was efficient in sealing the pores of the graft. Blood perfusion study showed that when hydrogel-coated grafts were exposed to blood for 30 min, they showed little affinity toward platelets or leukocytes. Hemolytic potential of PET was significantly reduced when it was coated with hydrogel. Improved adhesion and proliferation of endothelial cells were observed when PET grafts were coated with hydrogel. Results also showed that coating with hydrogel did not affect the burst strength of the PET graft.
When applied on the counter electrode of a dye-sensitized solar cell, functionalized graphene sheets with oxygen-containing sites perform comparably to platinum (conversion efficiencies of 5.0 and 5.5%, respectively, at 100 mW cm(-2) AM1.5G simulated light). To interpret the catalytic activity of functionalized graphene sheets toward the reduction of triiodide, we propose a new electrochemical impedance spectroscopy equivalent circuit that matches the observed spectra features to the appropriate phenomena. Using cyclic voltammetry, we also show that tuning our material by increasing the amount of oxygen-containing functional groups can improve its apparent catalytic activity. Furthermore, we demonstrate that a functionalized graphene sheet based ink can serve as a catalytic, flexible, electrically conductive counter electrode material.
A major question in transcription factor (TF) biology is why a TF binds to only a small fraction of motif eligible binding sites in the genome. Using the estrogen receptor-? as a model system, we sought to explicitly define parameters that determine TF-binding site selection. By examining 12 genetic and epigenetic parameters, we find that an energetically favorable estrogen response element (ERE) motif sequence, co-occupancy by the TF FOXA1, the presence of the H3K4me1 mark and an open chromatin configuration in the pre-ligand state provide specificity for ER binding. These factors can model estrogen-induced ER binding with high accuracy (ROC-AUC=0.95 and 0.88 using different genomic backgrounds). Moreover, when assessed in another estrogen-responsive cell line, this model was highly predictive for ER? binding (ROC-AUC=0.86). Variance in binding site selection between MCF-7 and T47D resides in sites with suboptimal ERE motifs, but modulated by the chromatin configuration. These results suggest a definable interplay between sequence motifs and local chromatin in selecting TF binding.
Genomes are organized into high-level three-dimensional structures, and DNA elements separated by long genomic distances can in principle interact functionally. Many transcription factors bind to regulatory DNA elements distant from gene promoters. Although distal binding sites have been shown to regulate transcription by long-range chromatin interactions at a few loci, chromatin interactions and their impact on transcription regulation have not been investigated in a genome-wide manner. Here we describe the development of a new strategy, chromatin interaction analysis by paired-end tag sequencing (ChIA-PET) for the de novo detection of global chromatin interactions, with which we have comprehensively mapped the chromatin interaction network bound by oestrogen receptor alpha (ER-alpha) in the human genome. We found that most high-confidence remote ER-alpha-binding sites are anchored at gene promoters through long-range chromatin interactions, suggesting that ER-alpha functions by extensive chromatin looping to bring genes together for coordinated transcriptional regulation. We propose that chromatin interactions constitute a primary mechanism for regulating transcription in mammalian genomes.
Woven polyethylene terephthalate (PET) fabric has been used in the construction of vascular grafts and sewing ring of prosthetic heart valves. In an effort to improve haemocompatibility and tissue response to PET fabric, a fluoropolymer, polyvinylidine fluoride (PVDF), was coated on PET fabric by dip coating technique. The coating was found to be uniform and no significant changes occurred on physical properties such as water permeability and burst strength. Cell culture cytotoxicity studies showed that coated PET was non-cytotoxic to L929 fibroblast cell lines. In vitro studies revealed that coating improved haemocompatibility of PET fabric material. Coating reduced platelet consumption of PET fabric by 50%. Upon surface modification leukocyte consumption of PET was reduced by 24%. About 60% reduction in partial thromboplastin time (PTT) observed when PET was coated with PVDF. Results of endothelial cell proliferation studies showed that surface coating did not have any substantial impact on cell proliferation. Overall results indicate that coating has potential to improve haemocompatibility of PET fabric without affecting its mechanical performance.
The synthesis and characterization of polyurethane (PU) with excellent radiopacity for medical and allied applications are reported. Bisphenol-A (BPA) was iodinated to obtain 4,4-isopropylidinedi-(2,6-diiodophenol) (IBPA) which was used as a chain extender for the preparation of a radiopaque PU. The PU was prepared by reacting 4,4-methylenebis(phenyl isocyanate) (MDI), poly(tetramethylene glycol) (PTMG) and IBPA in 2.2:1.2:1 molecular ratio and is characterized by infrared spectroscopy (IR), thermogravimetry (TGA), dynamic mechanical analysis (DMA), energy dispersive X-ray analysis (EDX), gel permeation chromatography (GPC) and X-radiography. X-ray images showed that the PU prepared using IBPA as the chain extender is highly radiopaque. An in vitro cytotoxicity test using L929 mouse fibroblast cells shows that the PU is non-cytotoxic. The outlined synthesis of a PU with radiocontrast properties opens up the possibility of synthesizing many different kinds of radiopaque PUs with desirable range of physical properties exploiting the versatility in their chemical synthesis.
A synthetic pathway has been constructed for the production of glucuronic and glucaric acids from glucose in Escherichia coli. Coexpression of the genes encoding myo-inositol-1-phosphate synthase (Ino1) from Saccharomyces cerevisiae and myo-inositol oxygenase (MIOX) from mice led to production of glucuronic acid through the intermediate myo-inositol. Glucuronic acid concentrations up to 0.3 g/liter were measured in the culture broth. The activity of MIOX was rate limiting, resulting in the accumulation of both myo-inositol and glucuronic acid as final products, in approximately equal concentrations. Inclusion of a third enzyme, uronate dehydrogenase (Udh) from Pseudomonas syringae, facilitated the conversion of glucuronic acid to glucaric acid. The activity of this recombinant enzyme was more than 2 orders of magnitude higher than that of Ino1 and MIOX and increased overall flux through the pathway such that glucaric acid concentrations in excess of 1 g/liter were observed. This represents a novel microbial system for the biological production of glucaric acid, a "top value-added chemical" from biomass.
Overt marking of BE in nonmainstream adult dialects of English is influenced by a number of linguistic constraints, including the structures person, number, tense, contractibility, and grammatical function. In the current study, the authors examined the effects of these constraints on overt marking of BE in children as a function of their nonmainstream English dialect and age.
A novel aerobic, non-motile, pleomorphic, Gram-negative and nitrogen-fixing bacterial strain, designated R5-392(T), was isolated from surface-sterilized root tissue of Jatropha curcas. The organism grew optimally at 30 °C in media containing 1 % (w/v) NaCl and at pH 6.0-8.0. The predominant ubiquinone was Q-10 and the major fatty acids were C18 : 1?7c/C18 : 1?6c, C16 : 0 and C19 : 0 cyclo ?8c. The DNA G+C content was 63.2 mol%. Analysis of the 16S rRNA gene sequence suggested that strain R5-392(T) is affiliated with the order Rhizobiales within the class Alphaproteobacteria and is most closely related to Pleomorphomonas oryzae F-7(T) (98.8 % similarity) and Pleomorphomonas koreensis Y9(T) (98.3 % similarity). Analysis of partial nifH gene sequences also revealed a monophyletic lineage within the class Alphaproteobacteria, and strain R5-392(T) was most closely related to P. oryzae F-7(T) (98 %). Highest nitrogenase activity was detected in the presence of low-level organic nitrogen or in the presence of nitrogenase co-factors (Fe/Mo) in N-free media. Phenotypic and chemotaxonomic data suggest that strain R5-392(T) represents a novel species within the genus Pleomorphomonas, for which the name Pleomorphomonas diazotrophica sp. nov. is proposed. The type strain is R5-392(T) ( = KACC 16233(T) = DSM 25022(T)).
Whereas the nature of healing reaction in skeletal muscle following implantation of biomaterial has been extensively studied, the extent of variation in cell phenotypes is poorly known. Here, we studied the phenotypic alteration of cell types following injury in skeletal muscle of rabbits implanted with a commonly used biomaterial, polyethylene terephthalate (PET) fabric. Following implantation, histomorphological features were studied after 1, 4, and 12 weeks. Routine objective histomorphological evaluation was supplemented with histochemistry for collagen and immunohistochemistry for proliferating cell nuclear antigen (PCNA), CD34, vimentin, and alpha smooth muscle actin (?-SMA). The extent of reaction was quantified. The foreign body giant cells were found to comprise subpopulations, based on the variation in vimentin detectability or the presence of differentially capable proliferating nuclei (PCNA positive). Many rhabdomyocytes adjacent to the implant were PCNA-positive and some of them showed CD34 positivity. Often, the rhabdomyocytes very near to implanted PET fabric assumed a myofibroblast phenotype as evidenced by vimentin and/or ?-SMA positivity at immunohistochemistry. Overall, the results suggested a phenotypic alteration of native cell types following implantation of PET fabric in rabbit skeletal muscle. Quantification of such cell types at the tissue-material interphase in relation to the deposition of collagen may be desirable during safety evaluation of biomaterials by histomorphology.
Synthesis and characterization of three different radiopaque thermoplastic polyurethane elastomers are reported. Radiopacity was introduced to the polyurethanes by incorporating an iodinated chain extender, namely, 4,4-isopropylidinedi-(2,6-diiodophenol) (IBPA), into the polymer chain during polyurethane synthesis. Radiopaque polyurethanes (RPUs) were synthesized by reacting 4,4-methylenebis(phenyl isocyanate) (MDI), IBPA, and three different diols. The polyols used for the synthesis were polypropylene glycol, polycaprolactone diol, and poly(hexamethylene carbonate) diol. RPUs were characterized by infrared spectroscopy, contact angle measurements, thermogravimetry, dynamic mechanical analysis, energy dispersive X-ray analysis, gel permeation chromatography, X-ray fluorescence spectroscopy, and X-radiography. X-ray images showed that all RPUs prepared using IBPA as the chain extender are highly radiopaque compared with an Aluminum wedge of equivalent thickness. Elemental analysis revealed that the polyurethanes contained 18-19% iodine in the polymer matrix. The RPUs developed have radiopacity equivalent to that of a polymer filled with 20 wt % barium sulfate. Results revealed that RPUs of wide range of properties may be produced by incorporating different diols as the soft chain segment. Cell culture cytotoxicity studies conducted using L929 cells by direct contact test and MTT assay proved that these RPUs are noncytotoxic in nature.
Babies born at lower gestational ages or smaller birthweights have a greater risk of poorer health in later life. Both the causes of these sub-optimal birth outcomes and the mechanism by which the effects are transmitted over decades are the subject of extensive study. We investigated whether a transcriptomic signature of either birthweight or gestational age could be detected in umbilical cord RNA.
Gemma is a database, analysis software system and web site for genomics data re-use and meta-analysis. Currently, Gemma contains analyzed data from over 3300 expression profiling studies, yielding hundreds of millions of differential expression results and coexpression patterns (correlated expression) for retrieval and visualization. With optional registration users can save their own data and securely share it with other users. Web services and integration with third-party resources further increase the scope of the tools, which include a Cytoscape plugin.
Several techniques for fabricating functionalized graphene sheet (FGS) electrodes were tested for catalytic performance in dye-sensitized solar cells (DSSCs). By using ethyl cellulose as a sacrificial binder, and partially thermolyzing it, we were able to create electrodes which exhibited lower effective charge transfer resistance (<1 ? cm(2)) than the thermally decomposed chloroplatinic acid electrodes traditionally used. This performance was achieved not only for the triiodide/iodide redox couple, but also for the two other major redox mediators used in DSSCs, based on cobalt and sulfur complexes, showing the versatility of the electrode. DSSCs using these FGS electrodes had efficiencies (?) equal to or higher than those using thermally decomposed chloroplatinic acid electrodes in each of the three major redox mediators: I (?(FGS) = 6.8%, ?(Pt) = 6.8%), Co (4.5%, 4.4%), S (3.5%, 2.0%). Through an analysis of the thermolysis of the binder and composite material, we determined that the high surface area of an electrode, as determined by nitrogen adsorption, is consistent with but not sufficient for high performing electrodes. Two other important considerations are that (i) enough residue remains in the composite to maintain structural stability and prevent restacking of FGSs upon the introduction of the solvent, and (ii) this residue must not disperse in the electrolyte.
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