A scalable solution-based approach is developed to controllably grow PPy ultrathin films on 2D MoS2 monolayers. When these sandwiched nanocomposites are utilized as supercapacitor electrodes, a record high specific capacitance, remarkable rate capability, and improved cycling stability are achieved, offering a feasible solution to create the next generation of energy-storage device with superior power density and energy density.

Here we explored a novel ZnO nanorod array template-assisted electrodeposition route to synthesize large-scale single-walled polypyrrole (PPy) nanotube arrays (NTAs) and multi-walled MnO(2)/PPy/MnO(2) NTAs. The structures of nanotubes, such as external and inner diameters, wall thicknesses, and lengths, can be well controlled by adjusting the diameters and lengths of ZnO nanorods and deposition time. The synthesized hybrid MnO(2)/PPy/MnO(2) triple-walled nanotube arrays (TNTAs) as electrodes showed high supercapacitive perporties, excellent long-term cycling stability, and high energy and power densities. The PPy layers in MnO(2)/PPy/MnO(2) TNTAs provide reliable electrical connections to MnO(2) shells and uniquely serve as highly conductive cores to support the redox reactions in the active two-double MnO(2) shells with highly electrolytic accessible surface area. The fabricated multi-walled NTAs allow high efficient utilization of electrode materials with facilitated transports of ions and electrons. The outstanding performance makes MnO(2)/PPy/MnO(2) TNTAs promising candidates for supercapacitor electrodes.

We evaluated the relationship between polymorphisms in the GSTM1 and GSTT1 genes and smoking status in a case-controlled study of a Korean population. The GSTM1 and GSTT1 genotypes were determined using a polymerase chain reaction (PCR)-based method and prognostic factors, such as staging and grading were evaluated for 126 bladder cancer patients, and 204 control subjects. Smoking represented a high-risk factor (odds ratio (OR)=4.8, 95% confidence interval (CI)=2.9-8.0) for the patients with bladder cancer. The frequency of GSTM1 null individuals was higher than in the controls, but the differences were not statistically significant (OR=1.56, 95% CI=2.9-8.0). For Korean subjects who smoked more than 1 pack of cigarettes per year (PPY), the increased risk of bladder cancer was associated with the GSTM1 null genotype (OR=0.5, 95% CI=0.3-0.9). Low-stage bladder tumors were more common among the GSTM1 null genotypes (OR=2.3; 95% CI=1.1-5.5). This study suggests that in Korean subjects the GSTM1 null genotype may be associated with increased risk for bladder cancer, in a manner that appears to depend upon smoking status. And also, in bladder cancer patients the GSTM1 null genotype appears to be associated with a poorer prognosis with low stage bladder tumors.

Diabetes mellitus is a challenging autoimmune disease. Biomedical researchers are currently exploring efficient and effective ways to solve this challenge. The potential of stem cell therapies for treating diabetes represents one of the major focuses of current research on diabetes treatment. Here, we have attempted to differentiate adult stem cells from umbilical cord blood-derived mesenchymal cells (UCB-MSC), Wharton's jelly-derived mesenchymal stem cells (WJ-MSC) and amniotic epithelial stem cells (AE-SC) into insulin-producing cells. The serum-free protocol developed in this study resulted in the differentiation of cells into definitive endoderm, pancreatic foregut, pancreatic endoderm and, finally, pancreatic endocrine cells, which expressed the marker genes SOX17, PDX1, NGN3, NKX6.1, INS, GCG, and PPY, respectively. Detection of the expression of the gap junction-related gene connexin-36 (CX36) using RT-PCR provided conclusive evidence for insulin-producing cell differentiation. In addition to this RT-PCR result, insulin and C-peptide protein were detected by immunohistochemistry and ELISA. Glucose stimulation test results showed that significantly greater amounts of C-peptide and insulin were released from differentiated cells than from undifferentiated cells. In conclusion, the methods investigated in this study can be considered an effective and efficient means of obtaining insulin-producing cells from adult stem cells within a week.

In this work, a new network nanocomposite composed of polypyrrole hydrogel (PPy hydrogel) loaded gold nanoparticles (AuNPs) was prepared. The PPy hydrogel was directly synthesized by mixing the pyrrole monomer and phytic acid, and the mixed solution can be gelated to form hydrogel at once. The three-dimensional network nanostructured PPy hydrogel not only provided a greater effective surface area for increasing the quantity of immobilized biomolecules and facilitated the transport of electrons and ions, but also exhibited an improved conductivity. Meanwhile, the electrodeposited AuNPs on the PPy hydrogel can further increase the specific surface area to capture a large amount of antibodies as well as improve the capability of electron transfer. The network PPy hydrogel/Au nanocomposites were successfully employed for the fabrication of a sensitive label-free amperometric immunosensor. Carcinoembryonic antigen (CEA) was used as a model protein. The proposed immunosensor exhibited a wide linear detection range from 1 fg mL(-1) to 200 ng mL(-1), and an ultralow limit of detection of 0.16 g mL(-1) (S/N = 3), and it also possessed good selectivity. Moreover, the detection of CEA in ten human serums showed satisfactory accuracy compared with the data determined by ELISA, indicating that the immunosensor provided potential application for clinical diagnosis.

The synthesis of a family of 4'-functionalized 5,5'-diaryl-2,2'-bipyridines (bpy*; 6a-6g) is reported. These ligands were reacted with the dimer [(ppy)(2)IrCl](2) (ppyH = 2-phenylpyridine) and afforded, after subsequent counterion exchange, a new series of luminescent cationic heteroleptic iridium(III) complexes, [(ppy)(2)Ir(bpy*)]PF(6) (8a-8g). These complexes were characterized by electrochemical and spectroscopic methods. The crystal structures of two of these complexes (8a and 8g) are reported. All of the complexes except for 8c and 8f exhibit intense and long-lived emission in both 2-MeTHF and ACN at 77 K and room temperature. The origin of this emission has been assigned by computational modeling to be an admixture of ligand-to-ligand charge-transfer [(3)LLCT; pi(ppy) ->> pi*(bpy*)] and metal-to-ligand charge-transfer [(3)MLCT; dpi(Ir) ->> pi*(bpy*)] excited states that are primarily composed of the former. The luminescent properties for 8a-8c are dependent upon the functionalization at the 4' position of the aryl substituents affixed to the diimine ligand, while those for 8d-8g are essentially independent because of an electronic decoupling of the aryls and bpy due to the substitution of o,o-dimethyl groups on the aryls, causing a near 90 degrees angle between the aryl and bipyridyl moieties. A combined density functional theory (DFT)/time-dependent DFT study was conducted in order to understand the origin of the transitions in the absorption and emission spectra and to predict accurately emission energies for these complexes.

Hypertension and diabetes mellitus are known to be frequently associated. The genetic dissection of diseases such as hypertension or diabetes mellitus is possible by using experimental crosses, which allow identification of loci influencing phenotypic traits (quantitative trait loci - QTLs). In this study the spontaneously hypertensive rat (SHR) and spontaneously diabetic, but normotensive rat (BB/OK) were crossed and the F2 population was analysed in order to search for QTLs on selected chromosomes (1, 10, 18) for blood pressure and some metabolic traits related to diabetes, renal function and hypertension. There were 3 regions found on chromosome 1 which showed linkage to blood pressure. The strongest evidence for linkage was observed between loci Igf2 and D1Mgh12. On chromosome 10 there was a QTL for blood pressure found between Ppy and Abp and on chromosome 18 there were three regions (Ttr-Grl, Tilp-Gja1, Olf-D18Mit9) with linkage to blood pressure. Since the 24 hr albumin and phosphate excretion correlated with blood pressure in F2 hybrids, the same regions were linked to both parameters. Region with linkage to serum concentrations of cholesterol (probably located beyond the terminal marker Ttr of the linkage group) were also found. The results of this study with a new F2(BB x SHR) population confirm the existence of previously described blood pressure loci (Sa and Bp2) and showed novel QTLs on chromosomes 1, 10 and 18.

Nanotechnology assists in the development of biocomposite nanofibrous scaffolds that can react positively to changes in the immediate cellular environment and stimulate specific regenerative events at molecular level to generate healthy tissues. Recently, electrospinning has gained huge momentum with greater accessibility of fabrication of composite, controlled and oriented nanofibers with sufficient porosity required for effective tissue regeneration. Current developments include the fabrication of nanofibrous scaffolds which can provide chemical, mechanical and biological signals to respond to the environmental stimuli. These nanofibers are fabricated by simple coating, blending of polymers/bioactive molecules or by surface modification methods. For obtaining optimized surface functionality, with specially designed architectures for the nanofibers (multi-layered, core-shell, aligned), electrospinning process has been modified and simultaneous 'electrospin-electrospraying' process is one of the most lately introduced technique in this perspective. Properties such as porosity, biodegradation and mechanical properties of composite electrospun nanofibers along with their utilization for nerve, cardiac, bone, skin, vascular and cartilage tissue engineering are discussed in this review. In order to locally deliver electrical stimulus and provide a physical template for cell proliferations, and to gain an external control on the level and duration of stimulation, electrically conducting polymeric nanofibers are also fabricated by electrospinning. Electrospun polypyrrole (PPy) and polyaniline (PAN) based scaffolds are the most extensively studied composite substrates for nerve and cardiac tissue engineering with or without electrical stimulations, and are discussed here. However, the major focus of ongoing and future research in regenerative medicine is to effectively exploit the pluripotent potential of Mesenchymal Stem Cell (MSC) differentiation on composite nanofibrous scaffolds for repair of organs.

The chromosomal region 17q12-q21 contains a gene (BRCA1) conferring susceptibility to early-onset familial breast and ovarian cancer. An 8000-rad radiation-reduced hybrid (RH) panel was constructed to provide a resource for long-range mapping of this region. A large fraction of the hybrids (approximately 90%) retained detectable human chromosome 17 sequences. The complete panel of 76 hybrids was scored for the presence or absence of 22 markers from this chromosomal region, including 14 cloned genes, seven microsatellite repeats, and one anonymous DNA segment. Statistical analysis of the marker retention data employing multipoint methods provided both comprehensive and framework maps of this chromosomal region, including distance estimates between adjacent markers. The comprehensive RH map includes 17 loci and spans 179 cRays(8000). Likelihood ratios of at least 1000:1 support the 10-locus framework order: cen-D17S250-ERBB2-(THRA1, TOP2A)-D17S855-PPY-D17S190-MTBT1-GP3A++ +-BTR-D17S588-tel. The order obtained from RH mapping, when used in conjunction with other methods, will be useful in linkage analysis of breast cancer families and will facilitate the development of a physical map of this region.

Electrically conductive and biologically active scaffolds are desirable for enhancing adhesion, proliferation and differentiation of a number of cell types such as neurons. Hence, the incorporation of neuroactive molecules into electroconductive polymers via a specific and stable method is essential for neuronal tissue engineering applications. Traditional conjugation approaches dramatically impair conductivities and/or stabilities of the scaffolds and ligands. In this study, we developed copolymers (PPy-NSE) of N-hydroxyl succinimidyl ester pyrrole and regular pyrrole, which can be immobilized with nerve growth factor (NGF) without significantly hindering electroconductivity. The presence of active ester groups was confirmed using reflectance infrared spectroscopy and X-ray photoelectron spectroscopy (XPS) from the copolymers prepared from different monomer compositions. We selected PPy-NSE(50) (polymerized from a 50 : 50 monomer ratio of pyrrole : pyrrole-NSE) for further modification with NGF because this copolymer retains good conductivity (approx. 8 S cm(-1)) and presents active ester groups for NGF immobilization. We tethered NGF on the PPy-NSE(50) surface, and found that PC12 cells extended neurites similarly to cells cultured in NGF-containing medium. XPS and enzyme-linked immunosorbent assay confirmed that NGF immobilized via the active ester on the PPy-NSE(50) film was stable for up to 5 days in phosphate-buffered saline solution. Also, application of an external electrical potential to NGF-immobilized PPy films did not cause a significant release of NGF nor reduce their neurotrophic activity. This novel scaffold, providing electroconductive and neurotrophic activities, has potential for neural applications, such as tissue engineering scaffolds and biosensors.

The distribution of the polypeptide which has an N-terminal tyrosine and a C-terminal tyrosine (PYY) - and pancreatic polypeptide (PP) - immunoreactive cells were investigated in the gut of the domestic fowl. PPY-immunoreactive cells were observed in the duodenum and jejunum. PP-immunoreactive cells were seen in the duodenum, jejunum, ileum and colon. Both PYY- and PP-immunoreactive cells were extended from the basal lamina to the gut lumen i.e. of open type. PYY-immunoreactive cells occurred mainly in the basal and middle portion of the villi. On the other hand, PP-immunoreactive cells were located mostly in the crepts. The occurrence of PYY-immunoreactive cells in the upper part of the small intestine is rather similar to that of amphibians and reptiles, than to that of mammals, where PYY-immunoreactive cells are located in the distal part of the small intestine and in the large intestine.

A functioning nanomachine in the form of a supramolecular nanovalve that opens and closes the orifices to molecular-sized pores and releases a small number of molecules on demand is reported. The nanovalve, which is used to open and close the nanocontainer, is a pseudorotaxane composed of two components-a long thread containing a 1,5-dioxnaphthalene donor unit, which is attached to the solid support, and the moving part, the tetracationic cyclophane acceptor/receptor, cyclobis(paraquat-p-phenylene), which controls access to the interior of the nanopore. The nanocontainer is made out of mesoporous silica by using a dip-coating method. Operating the nanovalve involves three steps: (i) filling the container, (ii) closing the valve, and (iii) opening the valve to release the contents of the container on demand. The tubular pores, which are approximately 2 nm wide, are filled with stable luminescent Ir(ppy)3 molecules by allowing them to diffuse into the open pores. The orifices are then closed by pseudorotaxane formation. An external reducing reagent (NaCNBH3) is used to effect dethreading of the pseudorotaxane so as to unlock the tubes and allow the guest molecules to be released. This nanovalve is a supramolecular machine consisting of a solid framework with moving parts capable of doing useful work.

Two-step electrochemical patterning methods have been employed to elaborate composite nanomaterials formed with multiwalled carbon nanotubes (MWCNTs) coated with polypyrrole (PPy) and redox PAMAM dendrimers. The nanomaterial has been demonstrated as a molecular transducer for electrochemical DNA detection. The nanocomposite MWCNTs-PPy has been formed by wrapping the PPy film on MWCNTs during electrochemical polymerization of pyrrole on the gold electrode. The MWCNTs-PPy layer was modified with PAMAM dendrimers of fourth generation (PAMAM G4) with covalent bonding by electro-oxidation method. Ferrocenyl groups were then attached to the surface as a redox marker. The electrochemical properties of the nanomaterial (MWCNTs-PPy-PAMAM-Fc) were studied using both square wave voltammetry and cyclic voltammetry to demonstrate efficient electron transfer. The nanomaterial shows high performance in the electrochemical detection of DNA hybridization leading to a variation in the electrochemical signal of ferrocene with a detection limit of 0.3 fM. Furthermore, the biosensor demonstrates ability for sensing DNA of rpoB gene of Mycobacterium tuberculosis in real PCR samples. Developed biosensor was suitable for detection of sequences with a single nucleotide polymorphism (SNP) T (TCG/TTG), responsible for resistance of M. tuberculosis to rifampicin drug, and discriminating them from wild-type samples without such mutation. This shows potential of such systems for further application in pathogens diagnostic and therapeutic purpose.

Electrical fields are known to interact with human cells. This principle has been explored to regulate cellular activities for bone tissue regeneration. In this work, Saos-2 cells were cultured on conductive scaffolds made of biodegradable poly(L-lactide) and the heparin-containing, electrically conducting polypyrrole (PPy/HE) to study their reaction to electrical stimulation (ES) mediated through such scaffolds. Both the duration and intensity of ES enhanced cell proliferation, generating a unique electrical intensity and temporal "window" within which osteoblast proliferation was upmodulated in contrast to the downmodulation or ineffectiveness in other ES regions. The favourable ES intensity (200 mV/mm) was further investigated in terms of the gene activation and protein production of two important osteoblast markers characterised by extracellular matrix maturation and mineralisation, that is alkaline phosphatase (ALP) and osteocalcin (OC). Both genes were found activated and the relevant protein production increased significantly following ES. In contrast, ES in the down-modulation region (400 mV/mm) suppressed the production of both ALP and OC. This work demonstrated that important osteoblast markers can be modulated with specific ES parameters mediated through conductive polymer substrates, providing a unique strategy for bone tissue engineering.

A visible-light-mediated procedure for the unprecedented trifluoromethylchlorosulfonylation of unactivated alkenes is presented. It uses [Cu(dap)2]Cl as catalyst, and contrasts with [Ru(bpy)3]Cl2, [Ir(ppy)2(dtbbpy)]PF6, or eosin Y that exclusively give rise to trifluoromethylchlorination of the same alkenes. It is assumed that [Cu(dap)2]Cl plays a dual role, that is, acting both as an electron transfer reagent as well as coordinating the reactants in the bond forming processes.

Pulmonary inflammation is a characteristic of many lung diseases. Increased levels of inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), and proinflammatory cytokines, such as interleukin-1beta (IL-1beta), tumor necrosis factor-alpha (TNF-alpha) and IL-8, have been correlated with lung inflammation. In this study, we used lipopolysaccharide (LPS) to induce iNOS, COX-2, and cytokines (TNF-alpha, IL-1beta, and IL-8) productions in human lung epithelial cells (A-549). Leaf of Eriobotrya japonica (Pi-Pa-Ye, PPY), a traditional Chinese medicine for the treatment of pulmonary inflammatory diseases, was capable of suppressing LPS-induced cytokine productions in a dose-dependent manner. Moreover, the suppression of PPY on the cytokine productions resulted from the inhibition of inhibitory kappaB-alpha phosphorylation and nuclear factor-kappaB (NF-kappaB) activation. Analysis of the anti-inflammatory effects of ursolic acid and oleanolic acid, the triterpene compounds present in PPY, showed that ursolic acid significantly inhibited LPS-induced IL-8 production, NF-kappaB activation, and iNOS mRNA expression, whereas oleanolic acid did not have these effects. In conclusion, our findings suggested the potential mechanisms of PPY and its active component, ursolic acid, in the treatment of pulmonary inflammation.

Here we describe a class of electric-conducting polymers that conduct electrons via the side chain π-π stacking. These polymers can be designed and synthesized with different chemical moieties to perform different functions, extremely suitable as a conductive polymer binder for lithium battery electrodes. A class of methacrylate polymers based on a polycyclic aromatic hydrocarbon side moiety, pyrene, was synthesized and applied as an electrode binder to fabricate a silicon (Si) electrode. The electron mobilities for PPy and PPyE are characterized as 1.9 × 10(-4) and 8.5 × 10(-4) cm(2) V(-1) s(-1), respectively. These electric conductive polymeric binders can maintain the electrode mechanical integrity and Si interface stability over a thousand cycles of charge and discharge. The as-assembled batteries exhibit a high capacity and excellent rate performance due to the self-assembled solid-state nanostructures of the conductive polymer binders. These pyrene-based methacrylate binders also enhance the stability of the solid electrolyte interphase (SEI) of a Si electrode over long-term cycling. The physical properties of this polymer are further tailored by incorporating ethylene oxide moieties at the side chains to enhance the adhesion and adjust swelling to improve the stability of the high loading Si electrode.

It is generally accepted that the neuropeptide Y (NPY) family of homologous peptides arose as a result of a series of gene duplication events. Recent advances in comparative genomics allow to formulate a hypothesis that explains, at least in part, the complexity of the family. Chromosome mapping studies reveal that the gene encoding PYY may have arisen from a common ancestral gene (termed NYY) in an ancient chromosomal duplication event that also involved the hox gene clusters. A tandem duplication of the PYY gene concomitant with or just before the emergence of tetrapods generated the PPY gene encoding PP. In the primate and ungulate lineages, the PYY-PPY gene cluster has undergone a more recent gene duplication event to create a PYY2-PPYPPY gene appears to have resulted in a relaxation of conservative pressure on the functional domain with the result that the amino acid sequences of tetrapod PYYs are more variable than the PYYs of jawed fish. Although the primary structure of PP has been quite strongly conserved in mammals, with the exception of the rodents, the extreme variability in the sequences of amphibian and reptilian PPs means that the peptide is a useful molecular marker to study the branching order in early tetrapod evolution

Eradicating subcutaneous bacterial infections remains a significant challenge. This work reports an injectable system of hollow microspheres (HMs) that can rapidly produce localized heat activated by near-infrared (NIR) light and control the release of an antibiotic via a "molecular switch" in their polymer shells, as a combination strategy for treating subcutaneous abscesses. The HMs have a shell of poly(d,l-lactic-co-glycolic acid) (PLGA) and an aqueous core that is comprised of vancomycin (Van) and polypyrrole nanoparticles (PPy NPs), which are photothermal agents. Experimental results demonstrate that the micro-HMs ensure efficiently the spatial stabilization of their encapsulated Van and PPy NPs at the injection site in mice with subcutaneous abscesses. Without NIR irradiation, the HMs elute a negligible drug concentration, but release substantially more when exposed to NIR light, suggesting that this system is suitable as a photothermally-responsive drug delivery system. The combination of photothermally-induced hyperthermia and antibiotic therapy with HMs increases cytotoxicity for bacteria in abscesses, to an extent that is greater than the sum of the two treatments alone, demonstrating a synergistic effect. This treatment platform may find other clinical applications, especially for localized hyperthermia-based cancer therapy.

Nanostructured conductive polymers can offer analogous environments for extracellular matrix and induce cellular responses by electric stimulation, however, such materials often lack mechanical strength and tend to collapse under small stresses. We prepared electrically conductive nanoporous materials by coating nanoporous cellulose gels (NCG) with polypyrrole (PPy) nanoparticles, which were synthesized in situ from pyrrole monomers supplied as vapor. The resulting NCG/PPy composite hydrogels were converted to aerogels by drying with supercritical CO2, giving a density of 0.41-0.53 g cm(-3), nitrogen adsorption surface areas of 264-303 m(2) g(-1), and high mechanical strength. The NCG/PPy composite hydrogels exhibited an electrical conductivity of up to 0.08 S cm(-1). In vitro studies showed that the incorporation of PPy into an NCG enhances the adhesion and proliferation of PC12 cells. Electrical stimulation demonstrated that PC12 cells attached and extended longer neurites when cultured on NCG/PPy composite gels with DBSA dopant. These materials are promising candidates for applications in nerve regeneration, carbon capture, catalyst supports, and many others.

This work describes development and optimization of a generic method for the immobilization of enzymes in chemically synthesized gold polypyrrole (Au-PPy) nanocomposite and their application in amperometric biosensors. Three enzyme systems have been used as model examples: cytochrome c, glucose oxidase and polyphenol oxidase. The synthesis and deposition of the nanocomposite was first optimized onto a glassy carbon electrode (GCE) and then, the optimum procedure was used for enzyme immobilization and subsequent fabrication of glucose and phenol biosensors. The resulting nanostructured polymer strongly adheres to the surface of the GCE electrode, has uniform distribution and is very stable. The method has proved to be an effective way for stable enzyme attachment while the presence of gold nanoparticles provides enhanced electrochemical activity; it needs very small amounts of pyrrole and enzyme and the Au-PPy matrix avoids enzyme leaking. The preparation conditions, Michaelis-Menten kinetics and analytical performance characteristics of the two biosensors are discussed. Optimization of the experimental parameters was performed with regard to pyrrole concentration, enzyme amount, pH and operating potential. These biosensors resulted in rapid, simple, and accurate measurement of glucose and phenol with high sensitivities (1.089 mA/M glucose and 497.1 mA/M phenol), low detection limits (2 x 10(-6)M glucose and 3 x 10(-8)M phenol) and fast response times (less than 10s). The biosensors showed an excellent operational stability (at least 100 assays) and reproducibility (R.S.D. of 1.36%).

One of the major benefits of a conductive PPy-based substrate is that the mediated electrical stimulation (ES) can be a stimulating factor to promote tissue regeneration. We cultured osteoblast-like Saos-2 cells on a conductive substrate made of biodegradable polylactide (95 wt%) and electrically conducting polypyrrole bioactivated with heparin (PPy/HE) (5 wt%). Using multi-well electrical cell culture plates, the effect of multiple ESs on osteoblast mineralization was investigated at various culture times. As ascertained by ARS, CPC and XPS analyses, the ES was able to promote osteoblast adhesion and growth, resulting in significantly higher calcium and phosphate content in the mineral deposition of the electrically stimulated membranes. Morphology, Ca/P ratio and crystalline structure demonstrated that the minerals on the conductive substrate surface were similar to those found on typical hydroxyapatite. ES also significantly upregulated the expression of the osteoblast-specific markers ALP, BMP2, Runx2 and OC. ES through a synthetic conductive polymer substrate therefore represents a vital option to promote bone regeneration.

SLiCE (Seamless Ligation Cloning Extract) is a novel cloning method that utilizes easy to generate bacterial cell extracts to assemble multiple DNA fragments into recombinant DNA molecules in a single in vitro recombination reaction. SLiCE overcomes the sequence limitations of traditional cloning methods, facilitates seamless cloning by recombining short end homologies (15-52 bp) with or without flanking heterologous sequences and provides an effective strategy for directional subcloning of DNA fragments from bacterial artificial chromosomes or other sources. SLiCE is highly cost-effective and demonstrates the versatility as a number of standard laboratory bacterial strains can serve as sources for SLiCE extract. We established a DH10B-derived E. coli strain expressing an optimized λ prophage Red recombination system, termed PPY, which facilitates SLiCE with very high efficiencies.

An electrochemical immunosensor for the common food pathogen Escherichia coli O157:H7 was developed. This novel immunosensor based on the PPy/AuNP/MWCNT/Chi hybrid bionanocomposite modified pencil graphite electrode (PGE). This hybrid bionanocomposite platform was modified with anti-E. coli O157:H7 monoclonal antibody. The prepared bionanocomposite platform and immunosensor was characterized by using cyclic voltammetry (CV). Under the optimum conditions, the results have shown the order of the preferential selectivity of the method is gram negative pathogenic species E. coli O157:H7. Concentrations of E. coli O157:H7 from 3×101 to 3×107cfu/mL could be detected. The detection limit was ∼30cfu/mL in PBS buffer. Briefly, we developed a high sensitive electrochemical immunosensor for specific detection of E. coli O157:H7 contamination with the use of sandwich assay evaluated in this study offered a reliable means of quantification of the bacteria. For the applications in food quality and safety control, our immunosensor showed reproducibility and stability.