bluuebunny/crossref_metadata_2025 huggingface dataset:
Solid‐Phase Synthesis of Molecularly Imprinted Polymer Nanoparticles with a Reusable Template–“Plastic Antibodies”
Authors: Poma Alessandro , Guerreiro Antonio , Whitcombe Michael J. , Piletska Elena V. , Turner Anthony P. F. , Piletsky Sergey A.
Published: January 2013

Molecularly imprinted polymers (MIPs) are generic alternatives to antibodies in sensors, diagnostics, and separations. To displace biomolecules without radical changes in infrastructure in device manufacture, MIPs should share their characteristics (solubility, size, specificity and affinity, localized binding domain) whilst maintaining the advantages of MIPs (low‐cost, short development time, and high stability) hence the interest in MIP nanoparticles. Herein, a reusable solid‐phase template approach is reported (fully compatible with automation) for the synthesis of MIP nanoparticles and their precise manufacture using a prototype automated UV photochemical reactor. Batches of nanoparticles (30–400 nm) with narrow size distributions imprinted with: melamine (d = 60 nm, Kd = 6.3 × 10−8 M), vancomycin (d = 250 nm, Kd = 3.4 × 10−9 M), a peptide (d = 350 nm, Kd = 4.8 × 10−8 M) and proteins have been produced. The instrument uses a column packed with glass beads, bearing the template. Process parameters are under computer control, requiring minimal manual intervention. For the first time, the reliable re‐use of molecular templates is demonstrated in the synthesis of MIPs (≥30 batches of nanoMIPs without loss of performance). NanoMIPs are produced template‐free and the solid‐phase acts both as template and affinity separation medium.
Phase Transformations and Structural Developments in the Radular Teeth of Cryptochiton Stelleri
Authors: Wang Qianqian , Nemoto Michiko , Li Dongsheng , Weaver James C. , Weden Brian , Stegemeier John , Bozhilov Krassimir N. , Wood Leslie R. , Milliron Garrett W. , Kim Christopher S. , DiMasi Elaine , Kisailus David
Published: January 2013

During mineralization, the hard outer magnetite‐containing shell of the radular teeth of Cryptochiton stelleri undergoes four distinct stages of structural and phase transformations: (i) the formation of a crystalline α‐chitin organic matrix that forms the structural framework of the non‐mineralized teeth, (ii) the templated synthesis of ferrihydrite crystal aggregates along these organic fibers, (iii) subsequent solid state phase transformation from ferrihydrite to magnetite, and (iv) progressive magnetite crystal growth to form continuous parallel rods within the mature teeth. The underlying α‐chitin organic matrix appears to influence magnetite crystal aggregate density and the diameter and curvature of the resulting rods, both of which likely play critical roles in determining the local mechanical properties of the mature radular teeth.
Striking intrafamilial phenotypic variability in Aicardi–Goutières syndrome associated with the recurrent Asian founder mutation in RNASEH2C
Authors: Vogt Julie , Agrawal Shakti , Ibrahim Zala , Southwood Taunton R. , Philip Sunny , MacPherson Lesley , Bhole Malini V. , Crow Yanick J. , Oley Christine
Published: January 2013

Aicardi–Goutières syndrome (AGS) is an encephalopathy of early childhood which is most commonly inherited as an autosomal recessive trait. The disorder demonstrates significant genetic heterogeneity with causative mutations in five genes identified to date. Although most patients with AGS experience a severe neonatal or infantile presentation, poor neurodevelopmental outcome and reduced survival, clinical variability in the onset and severity of the condition is being increasingly recognized. A later presentation with a more variable effect on development, morbidity and mortality has been particularly observed in association with mutations in SAMHD1 and RNASEH2B. In contrast, the recurrent c.205C > T (p.R69W) RNASEH2C Asian founder mutation has previously only been identified in children with a severe AGS phenotype. Here, to our knowledge, we present the first report of marked phenotypic variability in siblings both harboring this founder mutation in the homozygous state. In this family, one female child had a severe AGS phenotype with an onset in infancy and profound developmental delay, whilst an older sister was of completely normal intellect with a normal head circumference and was only diagnosed because of the presence of chilblains and a mild hemiplegia. An appreciation of intrafamilial phenotypic expression is important in the counseling of families considering prenatal diagnosis, and may also be relevant to the assessment of efficacy in future clinical trials. In addition, marked phenotypic variation raises the possibility that more mildly affected patients are not currently identified. © 2013 Wiley Periodicals, Inc.
Complex II deficiency—A case report and review of the literature
Authors: Jain‐Ghai Shailly , Cameron Jessie M. , Al Maawali Almundher , Blaser Susan , MacKay Nevena , Robinson Brian , Raiman Julian
Published: January 2013

Complex II deficiency is a rare cause of mitochondrial respiratory chain defects with a prevalence of 2–23%. It is exclusively nuclear encoded and functions in the citric acid cycle by oxidizing succinate to fumarate and in the mitochondrial electron transport chain (ETC) by transferring electrons to ubiquinone. Of the four subunits, SDHA and SDHB are catalytic and SDHC and SDHD are anchoring. Mutations in SDHA and SDHAF1 (assembly factor) have been found in patients with CII deficiency and a mitochondrial phenotype. We present a patient with CII deficiency with a previously undescribed phenotype of dilated cardiomyopathy, left ventricular noncompaction, failure to thrive, hypotonia, and developmental delay. Also, a comprehensive review of 36 cases published in the literature was undertaken. The results show that CII deficiency has a variable phenotype with no correlation with residual complex activity in muscle although the phenotype and enzyme activities are comparable within a family. For some, the condition was fatal in infancy, others had multisystem involvement and some had onset in adulthood with mild symptoms and normal cognition. Neurological involvement is most commonly observed and brain imaging commonly shows leukoencephalopathy, Leigh syndrome, or cerebellar atrophy. Mutations in SDHAF1 are associated with leukoencephalopathy. Other organ systems like heart, muscle, and eyes are only involved in about 50% of the cases but cardiomyopathy is associated with high mortality and morbidity. In some patients, riboflavin has provided clinical improvement. © 2013 Wiley Periodicals, Inc.
Singleton–Merten syndrome: An autosomal dominant disorder with variable expression
Authors: Feigenbaum Annette , Müller Christine , Yale Christopher , Kleinheinz Johannes , Jezewski Peter , Kehl Hans Gerd , MacDougall Mary , Rutsch Frank , Hennekam Raoul C.M.
Published: January 2013

In 1973, Singleton and Merten described two females with abnormal dentition, unique radiographic changes especially of the hands, and severe calcification and intimal weakening of the aortic arch and valve. Since then three additional cases with similar features have been reported and the diagnosis was suggested in another three individuals. We present an update of one case and the detailed clinical phenotype of six other cases with Singleton–Merten syndrome. The occurrence of the disorder in six members of two families and vertical male‐to‐male transmission indicate an autosomal dominant pattern of inheritance. Variability in phenotype, also within a single family, is significant. Core manifestations are marked aortic calcification, dental anomalies (delayed eruption and immature root formation of primarily the anterior permanent teeth, and early loss of permanent teeth due to short roots, acute root resorption, high caries, and aggressive alveolar bone loss), osteopenia and acro‐osteolysis, and to a lesser extend also glaucoma, psoriasis, muscle weakness, and joint laxity. Additional clinical characteristics described here include particular facial characteristics (high anterior hairline, broad forehead, smooth philtrum, thin upper vermillion) and abnormal joint and muscle ligaments. The cause and pathogenesis of this syndrome remain unknown. © 2013 Wiley Periodicals, Inc.
Molecular interaction of ctDNA and HSA with sulfadiazine sodium by multispectroscopic methods and molecular modeling
Authors: Geng Shaoguang , Liu Guosheng , Li Wei , Cui Fengling
Published: January 2013

Interactions of sulfadiazine sodium (SD‐Na) with calf thymus DNA (ctDNA) and human serum albumin (HSA) were studied using fluorescence spectroscopy, UV absorption spectroscopy and molecular modeling. The fluorescence experiments showed that the processes were static quenching. The results of UV spectra and molecular modeling of the interaction between SD‐Na and ctDNA indicated that the binding mode might be groove binding. In addition, the interaction of SD‐Na with HSA under simulative physiological conditions was also investigated. The binding constants (K) and the number of binding sites (n) at different temperatures (292, 302, 312 K) were 5.23 × 103 L/mol, 2.18; 4.50 × 103 L/mol, 2.35; and 4.08 × 103 L/mol, 2.47, respectively. Thermodynamic parameters including enthalpy change (ΔH) and entropy change (ΔS) were calculated, the results suggesting that hydrophobic force played a very important role in SD‐Na binding to HSA, which was in good agreement with the molecular modeling study. Moreover, the effect of SD‐Na on the conformation of HSA was analyzed using three‐dimensional fluorescence spectra. Copyright © 2013 John Wiley & Sons, Ltd.
Exceptionally E‐ and β‐Selective NHC–Cu‐Catalyzed Proto‐Silyl Additions to Terminal Alkynes and Site‐ and Enantioselective Proto‐Boryl Additions to the Resulting Vinylsilanes: Synthesis of Enantiomerically Enriched Vicinal and Geminal Borosilanes
Authors: Meng Fanke , Jang Hwanjong , Hoveyda Amir H.
Published: January 2013

An exceptionally site‐ and E‐selective catalytic method for preparation of Si‐containing alkenes through protosilylation of terminal alkynes is presented. Furthermore, the vinylsilanes obtained are used as substrates to generate vicinal or geminal borosilanes by another catalytic process; such products are derived from enantioselective protoborations of the Si‐substituted alkenes. All transformations are catalyzed by N‐heterocyclic carbene (NHC) copper complexes. Specifically, a commercially available imidazolinium salt, cheap CuCl (1.0 mol %) and Me2PhSi–B(pin), readily and inexpensively prepared in one vessel, are used to convert terminal alkynes to (E)‐β‐vinylsilanes efficiently (79–98 % yield) and in >98 % E and >98 % β‐selectivity. Vinylsilanes are converted to borosilanes with 5.0 mol % of a chiral NHC–Cu complex in 33–94 % yield and up to 98.5:1.5 enantiomeric ratio (e.r.). Alkyl‐substituted substrates afford vicinal borosilanes exclusively; aryl‐ and heteroaryl‐substituted alkenes deliver the geminal isomers preferentially. Different classes of chiral NHCs give rise to high enantioselectivities in the two sets of transformations: C1‐symmetric monodentate Cu complexes are most suitable for reactions of alkyl‐containing vinylsilanes and bidentate sulfonate‐bridged variants furnish the highest e.r. for substrates with an aryl substituent. Working models that account for the observed trends in selectivity are provided. Utility is demonstrated through application towards a formal enantioselective total synthesis of naturally occurring antibacterial agent bruguierol A.
Design, Synthesis, in vitro MAO‐B Inhibitory Evaluation, and Computational Studies of Some 6‐Nitrobenzothiazole‐Derived Semicarbazones
Authors: Tripathi Rati K. P. , Goshain Omprakash , Ayyannan Senthil Raja
Published: January 2013

Monoamine oxidase B (MAO‐B) is an important drug target for the treatment of neurological disorders. A series of 6‐nitrobenzothiazole‐derived semicarbazones were designed, synthesized, and evaluated as inhibitors of the rat brain MAO‐B isoenzyme. Most of the compounds were found to be potent inhibitors of MAO‐B, with IC50 values in the nanomolar to micromolar range. Molecular docking studies were performed with AutoDock 4.2 to deduce the affinity and binding mode of these inhibitors toward the MAO‐B active site. The free energies of binding (ΔG) and inhibition constants (Ki) of the docked compounds were calculated by the Lamarckian genetic algorithm (LGA) of AutoDock 4.2. Good correlations between the calculated and experimental results were obtained. 1‐[(4‐Chlorophenyl)(phenyl)methylene]‐4‐(6‐nitrobenzothiazol‐2‐yl)semicarbazide emerged as the lead MAO‐B inhibitor, with top ranking in both the experimental MAO‐B assay (IC50: 0.004±0.001 μM) and in computational docking studies (Ki: 1.08 μM). Binding mode analysis of potent inhibitors suggests that these compounds are well accommodated by the MAO‐B active site through stable hydrophobic and hydrogen bonding interactions. Interestingly, the 6‐nitrobenzothiazole moiety is stabilized in the substrate cavity with the aryl or diaryl residues extending up into the entrance cavity of the active site. According to our results, docking experiments could be an interesting approach for predicting the activity and binding interactions of this class of semicarbazones against MAO‐B. Thus, a binding site model consisting of three essential pharmacophoric features is proposed, and this can be used for the design of future MAO‐B inhibitors.
Overexpression of nerve growth factor by murine smooth muscle cells: Role of the p75 neurotrophin receptor on sympathetic and sensory sprouting
Authors: Petrie Casey N. , Smithson Laura J. , Crotty Anne‐Marie , Michalski Bernadeta , Fahnestock Margaret , Kawaja Michael D.
Published: January 2013

Elevating levels of nerve growth factor (NGF) can have pronounced effects on the survival and maintenance of distinct populations of neurons. We have generated a line of transgenic mice in which NGF is expressed under the control of the smooth muscle α‐actin promoter. These transgenic mice have augmented levels of NGF protein in the descending colon and urinary bladder, so these tissues display increased densities of NGF‐sensitive sympathetic efferents and sensory afferents. Here we provide a thorough examination of sympathetic and sensory axonal densities in the descending colon and urinary bladder of NGF transgenic mice with and without the expression of the p75 neurotrophin receptor (p75NTR). In response to elevated NGF levels, sympathetic axons (immunostained for tyrosine hydroxylase) undergo robust collateral sprouting in the descending colon and urinary bladder of adult transgenic mice (i.e., those tissues having smooth muscle cells); this sprouting is not augmented in the absence of p75NTR expression. As for sensory axons (immunostained for calcitonin gene‐related peptide) in the urinary bladders of transgenic mice, fibers undergo sprouting that is further increased in the absence of p75NTR expression. Sympathetic axons are also seen invading the sensory ganglia of transgenic mice; these fibers form perineuronal plexi around a subpopulation of sensory somata. Our results reveal that elevated levels of NGF in target tissues stimulate sympathetic and sensory axonal sprouting and that an absence of p75NTR by sensory afferents (but not by sympathetic efferents) leads to a further increase of terminal arborization in certain NGF‐rich peripheral tissues. J. Comp. Neurol. 521:2621–2643, 2013. © 2013 Wiley Periodicals, Inc.
Photoelectrochemical Scanning Droplet Cell Microscopy (PE‐SDCM)
Authors: Kollender Jan Philipp , Mardare Andrei Ionut , Hassel Achim Walter
Published: January 2013

Principles of localised photoelectrochemistry are summarised and an experimental approach is described that allows the performance of the most important photoelectrochemical experiments within a diameter of 100 μm. Various light sources, such as a continuum emitter with a monochromator, LEDs, and lasers are coupled into a multi‐mode fibre to illuminate a small spot that is wetted by the electrolyte from a capillary. Reference electrode, counter electrode, and optical fibre are installed in the capillary system. The performance of this system is demonstrated by photocurrent measurements on n‐doped Si and p‐doped Si as model substrates. A thickness‐graded aluminium thin film for partial shadowing on Si proves the applicability for material library investigations in combinatorial materials science. Further experiments demonstrate the possibility of electrical light chopping as well as impedance spectroscopy with subsequent Mott–Schottky analysis for the determination of charge‐carrier concentration and type, flat‐band potential, and inversion layer formation. Photoelectrochemical scanning droplet cell microscopy (PE‐SDCM) is an extremely versatile tool for the screening of water splitting photoelectrodes, the characterisation of photocatalysts, and high throughput characterisation of microgram amounts of new solar cell materials.
A threshold of 10% for myeloperoxidase by flow cytometry is valid to classify acute leukemia of ambiguous and myeloid origin
Authors: van den Ancker Willemijn , Westers Theresia M. , de Leeuw David C. , van der Veeken Yvonne F. C. M. , Loonen Anne , van Beckhoven Ernest , Ossenkoppele Gert J. , van de Loosdrecht Arjan A.
Published: January 2013

Background:

According to WHO 2008 guidelines, an important role is designated for cytoplasmic myeloperoxidase (cMPO) as measured by flow cytometry for classifying acute leukemia of myeloid or ambiguous origin (AML or MPAL). However, no threshold with respect to expression level and percentage positive cells is provided. Since the expression of solely cMPO can change the diagnosis from acute lymphoid leukemia into MPAL in the current WHO 2008, a consensus is needed for the cut‐off for cMPO.

Methods:

In this study, we investigated whether or not a cut‐off of 10% positivity for cMPO equally defines an acute leukemia as AML or MPAL as compared to a cut‐off for cMPO of 20% and compared this with results obtained for Sudan Black B (SBB) staining by cytomorphology.

Results:

Cell lineage‐defining markers and SBB staining were analyzed retrospectively in a cohort of 198 patients who presented with acute leukemia. Eight patients were positive for SBB (>3%), but were considered negative for cMPO (<10%); six patients were negative for SBB (≤3%) and positive for cMPO (≥10%) staining. In six patients, we found 10–20% cMPO positive leukemic cells. Five of these cases were SBB positive; the sixth patient showed a clear myeloid phenotype without positivity of any lymphoid marker. Using a 10% cut‐off instead of 20% would have changed diagnosis from ALL into MPAL in two patients; both cases were SBB positive by morphology.

Conclusion:

We conclude that a 10% cut‐off is a secure lower limit for cMPO expression and can be used independently from SBB expression. © 2013 International Clinical Cytometry Society
Drug‐Induced Convulsions in Nonclinical Safety Studies: Implication for Clinical Development
Authors: Elander Mikael
Published: January 2013

Preclinical Research
Preclinical Development Toxicology, Formulation Drug Delivery, Pharmacokinetics

The occurrence of convulsions in nonclinical safety studies during development of a new drug often complicates further development of the compound. Based on experiences with a previous development project at H. Lundbeck A/S, where convulsions were noted in animals, a structured approach to investigate possible mechanisms behind the convulsions is suggested. Because it is typically difficult to determine the exact reason for convulsions, it is important to focus on the assessment of possible human risk rather than finding the exact mechanism behind the convulsions. The suggested investigations include the following: calculation of safety margin and assessment of human risk, evaluation of data already captured in performed studies such as clinical chemistry data, description of clinical signs, batches of compound used, expert advice on activities that could enable further development of the compound, investigation of extended in vitro receptor profiles of parent compound and major metabolites, extended histopathology on brain sections, seizure threshold tests of parent compound and major metabolites (in vivo/in vitro), evaluation of electroencephalograms, and finally contact with authorities to discuss a plan for continued clinical development.
Regulation of muscle acetylcholine receptor turnover by β subunit tyrosine phosphorylation
Authors: Rudell John B. , Ferns Michael J.
Published: January 2013

At the neuromuscular junction (NMJ), the postsynaptic localization of muscle acetylcholine receptor (AChR) is regulated by neural signals and occurs via several processes including metabolic stabilization of the receptor. However, the molecular mechanisms that influence receptor stability remain poorly defined. Here, we show that neural agrin and the tyrosine phosphatase inhibitor, pervanadate slow the degradation of surface receptor in cultured muscle cells. Their action is mediated by tyrosine phosphorylation of the AChR β subunit, as agrin and pervandate had no effect on receptor half‐life in AChR‐β3F/3F muscle cells, which have targeted mutations of the β subunit cytoplasmic tyrosines. Moreover, in wild type AChR‐β3Y muscle cells, we found a linear relationship between average receptor half‐life and the percentage of AChR with phosphorylated β subunit, with half‐lives of 12.7 and 23 h for nonphosphorylated and phosphorylated receptor, respectively. Surprisingly, pervanadate increased receptor half‐life in AChR‐β3Y myotubes in the absence of clustering, and agrin failed to increase receptor half‐life in AChR‐β3F/3F myotubes even in the presence of clustering. The metabolic stabilization of the AChR was mediated specifically by phosphorylation of βY390 as mutation of this residue abolished β subunit phosphorylation but did not affect δ subunit phosphorylation. Receptor stabilization also led to higher receptor levels, as agrin increased surface AChR by 30% in AChR‐β3Y but not AChR‐β3F/3F myotubes. Together, these findings identify an unexpected role for agrin‐induced phosphorylation of βY390 in downregulating AChR turnover. This likely stabilizes AChR at developing synapses, and contributes to the extended half‐life of AChR at adult NMJs. © 2013 Wiley Periodicals, Inc. Develop Neurobiol 73: 399–410, 2013
Small creatures use small RNAs to direct antiviral defenses
Authors: Sabin Leah R. , Cherry Sara
Published: January 2013

Antiviral RNA silencing has been recognized as an important defense mechanism in arthropods against RNA viruses. However, the role of this pathway in DNA virus infection remains largely unexplored. A report in this issue of the European Journal of Immunology provides new insight into the role of RNA silencing in antiviral defense against DNA viruses. Huang and Zhang [Eur. J. Immunol. 2013. 137–146] found that the dsDNA virus white spot syndrome virus, an agriculturally important pathogen of shrimp, is targeted by the shrimp RNA‐silencing machinery via the production of virus‐derived siRNAs. Furthermore, the authors show that the RNA‐silencing pathway, and crucially, Dicer‐2, is important for restricting viral infection. This study provides novel insights not only into shrimp antiviral defenses but also potentially into antiviral immunity against DNA viruses in a larger spectrum of hosts, as discussed in this Commentary. Furthermore, this study may contribute to the future development of immune‐based therapeutics to combat viral pathogens, not only in aquaculture, but also in insect vectors of human diseases.
Relevance of minor components stability in commercial olive oil quality during the market period
Authors: Fregapane Giuseppe , Gómez‐Rico Aurora , Inarejos Antonio M. , Salvador M. Desamparados
Published: January 2013

The aim of this research was to study the relevance of the stability of minor components in commercial Spanish extra virgin and olive oil (OO) categories, particularly with respect to the individual phenolic compounds, tocopherols (vitamin E) and volatile compounds, during the market period (12 months). The study was carried out classifying several commercial olive oils (n = 39) according to their content in phenolic compounds into three different groups: extra virgin olive oils with high (>290 mg/kg; HP‐VOO) and low (<210 mg/kg; LP‐VOO) phenolic level and OO, due to the impact of biophenols on oxidative stability, sensory properties and nutritional aspects. After a market period of 1 year, the α‐tocopherol content fell significantly (25–30% of its initial content) in the commercial oils. While the decrease in total phenolics and o‐diphenols was not statistically significant, a relevant increase (33% on average) in simple phenolics (hytroxytyrosol and tyrosol) was observed in all the studied samples, suggesting its use as freshness or ageing marker. Throughout the market period, a low oxidation rate and therefore a high stability of the products was observed. Moreover, the results found in the study suggest that a higher market price OO is apparently related to better quality and process control, giving a superior final product in terms of its content of minor components related to organoleptic properties.

Practical applications: The quality of virgin olive oil – mainly its sensory characteristics and nutritional value – is directly related to its content in minor components, in particular phenolics and volatile compounds, and therefore to their stability during the market period. Nevertheless, this information is not available to consumers. As a consequence, it is very important for both producers and consumers to clarify and establish the real relevance of the stability of minor components in commercial olive oil categories during their shelf‐life: how long they will keep their stability, organoleptic and nutritional characteristics. This information would highlight the individual quality properties of a specific oil product, beyond the sometimes misleading commercial category.
Deposition Strategies for Osmium/Enzyme Films on Gold Electrode Based Sensing Arrays
Authors: McKenzie Jennifer R. , Zhang Chengxiao , Li Chen‐Zhong
Published: January 2013

Multi‐analyte real‐time interrogation of cellular activity allows for the potential discovery of novel insights into disease. In this report, the addition of electrochemical biosensors to a previously developed platform utilizing Au microfabricated electrodes was explored. Glucose oxidase was immobilized at the electrode surface with an osmium redox polymer, using hand‐casting and electrodeposition techniques, allowing for the first comparison of deposition techniques at a Au microfabricated array. This preliminary work is the first step toward the goal of creating a multi‐analyte array for cellular analysis.
An aspect ratio agglomeration multigrid for unstructured grids
Authors: Li Zongzhe , Wang Zhenghua , Cao Wei , Yao Lu
Published: January 2013

A robust aspect ratio‐based agglomeration algorithm to generate high quality of coarse grids for unstructured and hybrid grids is proposed in this paper. The algorithm focuses on multigrid techniques for the numerical solution of Euler and Navier–Stokes equations, which conform to cell‐centered finite volume special discretization scheme, combines vertex‐based isotropic agglomeration and cell‐based directional agglomeration to yield large increases in convergence rates. Aspect ratio is used as fusing weight to capture the degree of cell convexity and give an indication of cell stretching. Agglomeration front queue is established to propagate inward from the boundaries, which stores isotropic vertex and also high‐stretched cell marked with different flag according to aspect ratio. We conduct the present method to solve Euler and Navier–Stokes equations on unstructured and hybrid grids and compare the results with single grid as well as MGridGen, which shows that the present method is efficient in reducing computational time for large‐scale system equations. Copyright © 2013 John Wiley & Sons, Ltd.
Ingenol mebutate: a novel topical agent for actinic keratosis
Authors: Morton Colin A
Published: January 2013

Ingenol mebutate is a once‐daily topical treatment for actinic keratosis (AK) that is applied for only two or three days depending on body site. It has recently been approved by the FDA for the treatment of AK on the face, scalp, trunk and extremities and recommended for approval by the CHMP in September 2012. In this drug profile, the author reviews its mode of action, efficacy and tolerability and comments on the drug's likely place in practice. Copyright © 2013 John Wiley & Sons, Ltd.
Detailed Dynamic Modeling of a Molten Carbonate Fuel Cell Stack with Indirect Internal Reformers
Authors: Kim H. , Cho J. H. , Lee K. S.
Published: January 2013

A detailed dynamic model of a molten carbonate fuel cell (MCFC) stack that can predict the distribution of process variables over the cell surface and along the stacking direction is presented. The model describes a repeating unit of the stack, which is composed of eight cells and one indirect internal reformer (IIR). The set of partial differential equations for the MCFC model is reduced to a set of ordinary differential equations through discretization using the cubic spline collocation method and the finite difference method. The steady‐state and dynamic behaviors of the eight‐cell stack model are compared with those of the one‐cell model, which has been commonly used in previous studies on MCFC dynamics, and three‐cell aggregate model. The distribution of the process variables along the stacking direction cannot be disregarded, and thus more detailed model should be used for the accurate reproduction of MCFC dynamics.
Proton Mobility in Sulfonated PolyEtherEtherKetone (SPEEK): Influence of Thermal Crosslinking and Annealing
Authors: Knauth P. , Pasquini L. , Maranesi B. , Pelzer K. , Polini R. , Di Vona M. L.
Published: January 2013

Sulfonated PolyEtherEtherKetone (SPEEK) was thermally treated at 180 and 140 °C to study the effects of polymer cross‐linking and annealing on the water uptake and proton conductivity. The kinetics of the cross‐linking reaction can be quantitatively described with an activation energy of (100 ± 2) kJ mol–1. The proton mobility u(H+) depends on the proton concentration c(H+) in excellent agreement with the power law u(H+) ∝ c(H+)–3 observed above the percolation threshold of hydrated nanometric channels. The proton mobility can be described by porosity and tortuosity as phenomenological parameters. Polymer annealing reduces the free volume and porosity of membranes. The proton conductivity can be modulated by changing the hydration number.
Electrocatalytic Activity and Preparation of Pt29Ru51P15/CNTs Catalyst Using the Assistance of Thioglycolic Acid Functionalization
Authors: Xie J. , Xia D.‐G.
Published: January 2013

Pt29Ru51P15 particles with a uniform size distribution of 2 nm supported on multi‐walled carbon nanotubes (CNTs) have been prepared with the assistance of thioglycolic acid functionalization using NaH2PO2 · H2O as a reducing agent. The Pt29Ru51P15/CNTs has been characterized by transmission electron microscopy (TEM), X‐ray diffraction (XRD), and X‐ray photoelectron spectroscopy (XPS). Its electrochemical properties have been examined by cyclic voltammetry (CV) and chronoamperometry (CA). The results show that Pt29Ru51P15/CNTs has a much narrower particle size distribution with no formation of aggregates, a greater electrochemically active surface area, and higher electrocatalytic activity for the electro‐oxidation of methanol
Dopamine D3 receptor activation promotes neural stem/progenitor cell proliferation through AKT and ERK1/2 pathways and expands type‐B and ‐C cells in adult subventricular zone
Authors: Lao Chu Lan , Lu Chin‐Song , Chen Jin‐Chung
Published: January 2013

The neurotransmitter dopamine acts on the subventricular zone (SVZ) to regulate both prenatal and postnatal neurogenesis, in particular through D3 receptor (D3R) subtype. In this study, we explored the cellular mechanism(s) underlying D3R‐mediated cell proliferation and tested if systemic delivery of a D3R agonist would induce SVZ multipotent neural stem/precursor cell (NSC/NPC) proliferation in vivo. We found that treatment with the D3R agonist, 7‐OH‐DPAT, enhances cell proliferation in a dose‐dependent manner in cultured SVZ neurospheres from wild‐type, but not D3R knock‐out mice. Furthermore, D3R activation also stimulates S‐phase and enhances mRNA and protein levels of cyclin D1 in wild‐type neurospheres, a process which requires cellular Akt and ERK1/2 signaling. Moreover, chronic treatment with low dose 7‐OH‐DAPT in vivo increases BrdU+ cell numbers in the adult SVZ, but this effect was not seen in D3R KO mice. Additionally, we probed the cell type specificity of D3R agonist‐mediated cell proliferation. We found that in adult SVZ, GFAP+ astrocytes, type‐B GFAP+/nestin+ and type‐C EGF receptor (EGFR+)/nestin+ cells express D3R mRNA, but type‐A Doublecortin (Dcx)+ neuroblasts do not. Using flow cytometry and immunofluorescence, we demonstrated that D3R activation increases GFAP+ type‐B and EGFR+ type‐C cell numbers, and the newly divided Dcx+ type‐A cells. However, BrdU+/Dcx+ cell numbers were decreased in D3R KO mice compared to wildtype, suggesting that D3R maintains constitutive NSC/NPCs population in the adult SVZ. Overall, we demonstrate that D3R activation induces NSC/NPC proliferation through Akt and ERK1/2 signaling and increases the numbers of type‐B and ‐C NSC/NPCs in the adult SVZ. © 2013 Wiley Periodicals, Inc.
Induction of atypical EAE mediated by transgenic production of IL‐6 in astrocytes in the absence of systemic IL‐6
Authors: Giralt Mercedes , Ramos Raquel , Quintana Albert , Ferrer Beatriz , Erta Maria , Castro‐Freire Marco , Comes Gemma , Sanz Elisenda , Unzeta Mercedes , Pifarré Paula , García Agustina , Campbell Iain L. , Hidalgo Juan
Published: January 2013

Interleukin (IL)‐6 is crucial for the induction of many murine models of autoimmunity including experimental autoimmune encephalomyelitis (EAE), an animal model of multiple sclerosis. While IL‐6‐deficient mice (IL‐6 KO) are resistant to EAE, we showed previously that in transgenic mice with astrocyte‐targeted production of IL‐6‐restricted to the cerebellum (GFAP‐IL6), EAE induced with MOG35–55 was redirected away from the spinal cord to the cerebellum. To further establish the importance of IL‐6 produced in the central nervous system, we have generated mice producing IL‐6 essentially only in the brain by crossing the GFAP‐IL6 mice with IL‐6 KO mice. Interestingly, GFAP‐IL6‐IL‐6 KO mice showed a milder but almost identical phenotype as the GFAP‐IL6 mice, which correlated with a lower load of inflammatory cells and decreased microglial reactivity. These results indicate that not only is cerebellar IL‐6 production and eventual leakage into the peripheral compartment the dominating factor controlling this type of EAE but that it can also facilitate induction of autoimmunity in the absence of normal systemic IL‐6 production. © 2013 Wiley Periodicals, Inc.
Microglial activation and expression of immune‐related genes in a rat ex vivo nervous system model after infection with Listeria monocytogenes
Authors: Remuzgo‐MartÍnez Sara , Pilares‐Ortega Lilian , Icardo Jose M. , ValdizÁn Elsa Maria , Vargas VerÓnica InÉs , Pazos Ángel , Ramos‐Vivas Jose
Published: January 2013

A wide variety of microorganisms has previously been identified as causes of brain infection. Among them, Listeria monocytogenes has a particular tropism for the central nervous system. To gain knowledge about the immune response elicited by L. monocytogenes in the brain, we used a rat ex vivo organotypic nervous system culture as a model for Listeria infection. Scanning electron microscopy (SEM) revealed that activated microglial cells showing a typical amoeboid morphology are quickly recruited to the surface of the explants after the infection. After bacterial engulfment, these cells appear to act as Trojan horses, releasing the engulfed bacteria inside the brain tissue. We describe cycles of microglial phagocytosis, necrotic cell death and the subsequent removal of cell debris for the first time. Furthermore, we used this ex vivo model to assess the expression profiles of immune relevant genes up to 24 h postinfection by means of q‐PCR‐arrays, finding that a number of inflammation‐promoting genes are upregulated. Shortly after infection by L. monocytogenes, upregulated genes were those that encoded molecules involved in Th1 responses, being the Ccl2 chemokine and members of the interleukin1‐β family the most abundant immunomodulatory signals expressed. After 5 h of infection, L. monocytogenes caused a substantial increase in the expression of TLR1 and TLR2 genes, as well as in several downstream genes of the TLR signaling pathways. © 2013 Wiley Periodicals, Inc.
Efficient drug screening and gene correction for treating liver disease using patient-specific stem cells
Authors: Choi Su Mi , Kim Yonghak , Shim Joong Sup , Park Joon Tae , Wang Rui-Hong , Leach Steven D. , Liu Jun O. , Deng Chuxia , Ye Zhaohui , Jang Yoon-Young
Published: January 2013

Patient-specific induced pluripotent stem cells (iPSCs) represent a potential source for developing novel drug and cell therapies. Although increasing numbers of disease-specific iPSCs have been generated, there has been limited progress in iPSC-based drug screening/discovery for liver diseases, and the low gene-targeting efficiency in human iPSCs warrants further improvement. Using iPSC lines from patients with alpha-1 antitrypsin (AAT) deficiency, for which there is currently no drug or gene therapy available, we established a platform to discover new drug candidates and correct disease-causing mutation with a high efficiency. A high-throughput format screening assay, based on our hepatic differentiation protocol, was implemented to facilitate automated quantification of cellular AAT accumulation using a 96-well immunofluorescence reader. To expedite the eventual application of lead compounds to patients, we conducted drug screening utilizing our established library of clinical compounds (the Johns Hopkins Drug Library) with extensive safety profiles. Through a blind large-scale drug screening, five clinical drugs were identified to reduce AAT accumulation in diverse patient iPSC-derived hepatocyte-like cells. In addition, using the recently developed transcription activator-like effector nuclease technology, we achieved high gene-targeting efficiency in AAT-deficiency patient iPSCs with 25%-33% of the clones demonstrating simultaneous targeting at both diseased alleles. The hepatocyte-like cells derived from the gene-corrected iPSCs were functional without the mutant AAT accumulation. This highly efficient and cost-effective targeting technology will broadly benefit both basic and translational applications.

Conclusions

: Our results demonstrated the feasibility of effective large-scale drug screening using an iPSC-based disease model and highly robust gene targeting in human iPSCs, both of which are critical for translating the iPSC technology into novel therapies for untreatable diseases.
Hepatic reticuloendothelial system cell iron deposition is associated with increased apoptosis in nonalcoholic fatty liver disease
Authors: Maliken Bryan D. , Nelson James E. , Klintworth Heather M. , Beauchamp Mary , Yeh Matthew M. , Kowdley Kris V.
Published: January 2013

The aim of this study was to examine the relationship between the presence of hepatic iron deposition, apoptosis, histologic features, and serum markers of oxidative stress (OS) and cell death in nonalcoholic fatty liver disease (NAFLD). Clinical, biochemical, metabolic, and independent histopathologic assessment was conducted in 83 unselected patients with biopsy-proven NAFLD from a single center. Apoptosis and necrosis in serum was quantified using serum cytokeratin 18 (CK18) M30 and M65 enzyme-linked immunosorbent assays and in liver by terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) staining

in situ

. Serum malondialdehyde (MDA) and thioredoxin-1 (Trx1) levels were measured to evaluate OS. Presence of reticuloendothelial system (RES) cell iron in the liver was associated with nonalcoholic steatohepatitis (

P

< 0.05) and increased hepatic TUNEL staining (

P

= 0.02), as well as increased serum levels of apoptosis-specific (M30;

P

= 0.013) and total (M65;

P

= 0.006) CK18 fragments, higher MDA (

P

= 0.002) and lower antioxidant Trx1 levels (

P

= 0.012), compared to patients without stainable hepatic iron. NAFLD patients with a hepatocellular (HC) iron staining pattern also had increased serum MDA (

P

= 0.006), but not M30 CK18 levels or TUNEL staining, compared to subjects without stainable hepatic iron. Patients with iron deposition limited to hepatocytes had a lower proportion of apoptosis-specific M30 fragments relative to total M65 CK18 levels (37% versus ≤25%;

P

< 0.05).

Conclusions

: Presence of iron in liver RES cells is associated with NASH, increased apoptosis, and increased OS. HC iron deposition in NAFLD is also associated with OS and may promote hepatocyte necrosis in this disease.
Plasma interferon-gamma-inducible protein-10 (IP-10) levels during acute hepatitis C virus infection
Authors: Grebely Jason , Feld Jordan J. , Applegate Tanya , Matthews Gail V. , Hellard Margaret , Sherker Alana , Petoumenos Kathy , Zang Geng , Shaw Ineke , Yeung Barbara , George Jacob , Teutsch Suzy , Kaldor John M. , Cherepanov Vera , Bruneau Julie , Shoukry Naglaa H. , Lloyd Andrew R. , Dore Gregory J.
Published: January 2013

Systemic levels of interferon-gamma-inducible protein-10 (IP-10) are predictive of treatment-induced clearance in chronic hepatitis C virus (HCV). In the present study, factors associated with plasma IP-10 levels at the time of acute HCV detection and the association between IP-10 levels and spontaneous clearance were assessed in three cohorts of acute HCV infection. Among 299 individuals, 245 (181 male, 47 human immunodeficiency virus-positive [HIV+]) were HCV RNA+ at acute HCV detection. In adjusted analysis, factors independently associated with IP-10 levels ≥150 pg/mL (median level) included HCV RNA levels >6 log IU/mL, HIV coinfection and non-Aboriginal ethnicity. Among 245 HCV RNA+ at acute HCV detection, 214 were untreated (n = 137) or had persistent infection (infection duration ≥26 weeks) at treatment initiation (n = 77). Spontaneous clearance occurred in 14% (29 of 214). Individuals without spontaneous clearance had significantly higher mean plasma IP-10 levels at the time of acute HCV detection than those with clearance (248 ± 32 versus 142 ± 22 pg/mL,

P

= 0.008). The proportion of individuals with spontaneous clearance was 0% (0 of 22,

P

= 0.048) and 16% (27 of 165) and in those with and without plasma IP-10 levels ≥380 pg/mL. In adjusted analyses, favorable IL28B genotype was associated with spontaneous clearance, while higher HCV RNA level was independently associated with lower odds of spontaneous clearance.

Conclusion

: High IP-10 levels at acute HCV detection were associated with failure to spontaneously clear HCV. Patients with acute HCV and high baseline IP-10 levels, particularly >380 pg/mL, should be considered for early therapeutic intervention, and those with low levels should defer therapy for potential spontaneous clearance.
Soluble receptor for advanced glycation end products and risk of liver cancer
Authors: Moy Kristin A. , Jiao Li , Freedman Neal D. , Weinstein Stephanie J. , Sinha Rashmi , Virtamo Jarmo , Albanes Demetrius , Stolzenberg-Solomon Rachael Z.
Published: January 2013

Binding of advanced glycation end products (AGEs) to their receptor (RAGE) increases oxidative stress and inflammation and may be involved in liver injury and subsequent carcinogenesis. Soluble RAGE (sRAGE) may neutralize the effects mediated by the AGE/RAGE complex. Epidemiologic studies examining sRAGE or AGEs in association with liver cancer are lacking. We examined the associations between prediagnostic serum concentrations of sRAGE or Nε-(carboxymethyl)-lysine (CML)-AGE and hepatocellular carcinoma in a case-cohort study within a cohort of 29,133 Finnish male smokers who completed questionnaires and provided a fasting blood sample between 1985 and 1988. During follow-up beginning 5 years after enrollment through April 2006, 145 liver cancers occurred. Serum concentrations of sRAGE, CML-AGE, glucose, and insulin were measured in case subjects and 485 randomly sampled cohort participants. Chronic hepatitis B virus (HBV) and hepatitis C virus (HCV) were available in most cases and in a subset of the study population. Weighted Cox proportional hazards regression was used to calculate relative risks (RR) and 95% confidence intervals (CI) adjusted for age, years of smoking, and body mass index. sRAGE and CML-AGE concentrations were inversely associated with liver cancer. Further adjustment for glucose and insulin or exclusion of case subjects with chronic HBV or HCV did not change the associations.

Conclusion

: Our results support the hypothesis that sRAGE is inversely associated with liver cancer. The findings need confirmation, particularly in populations that include women and nonsmokers.
Application of L‐moments and Bayesian inference for low‐flow regionalization in Sefidroud basin, Iran
Authors: Dodangeh Esmaeel , Soltani Saeed , Sarhadi Ali , Shiau Jenq‐Tzong
Published: January 2013

Reliable estimation of low flows at ungauged catchments is one of the major challenges in water‐resources planning and management. This study aims at providing at‐site and ungauged sites low‐flow frequency analysis using regionalization approach. A two‐stage delineating homogeneous region is proposed in this study. Clustering sites with similar low‐flow L‐moment ratios is initially conducted, and L‐moment‐based discordancy and heterogeneity measures are then used to detect unusual sites. Based on the goodness‐of‐fit test statistic, the best‐fit regional model is identified in each hydrologically homogeneous region. The relationship between mean annual 7‐day minimum flow and hydro‐geomorphic characteristics is also constructed in each homogeneous region associated with the derived regional model for estimating various low‐flow quantiles at ungauged sites. Uncertainty analysis of model parameters and low‐flow estimations is carried out using the Bayesian inference. Applied in Sefidroud basin located in northwestern Iran, two hydrologically homogeneous regions are identified, i.e. the east and west regions. The best‐fit regional model for the east and west regions are generalized logistic and Pearson type III distributions, respectively. The results show that the proposed approach provides reasonably good accuracy for at‐site as well as ungauged‐site frequency analysis. Besides, interval estimations for model parameters and low flows provide uncertainty information, and the results indicate that Bayesian confidence intervals are significantly reduced when comparing with the outcomes of conventionalt‐distribution method. Copyright © 2013 John Wiley & Sons, Ltd.