Isolation and characterization of a monoclonal anti-quadruplex DNA antibody from autoimmune "viable motheaten" mice

A cell line that produces an autoantibody specific for DNA quadruplex structures has been isolated and cloned from a hybridoma library derived from 3-month-old nonimmunized autoimmune, immunodeficient "viable motheaten" mice. This antibody has been tested extensively in vitro and found to bind specifically to DNA quadruplex structures formed by two biologically relevant sequence motifs. Scatchard and nonlinear regression analyses using both one- and two-site models were used to derive association constants for the antibody-DNA binding reactions. In both cases, quadruplexes had higher association constants than triplex and duplex molecules. The anti-quadruplex antibody binds to the quadruplex formed by the promoter-region-derived oligonucleotide d(CGCG4GCG) (Ka = 3.3 x 10(6) M-1), and has enhanced affinity for telomere-derived quadruplexes formed by the oligonucleotides d(TG4) and d(T2G4T2G4T2G4T2G4) (Ka = 5.38 x 10(6) and 1.66 x 10(7) M-1, respectively). The antibody binds both types of quadruplexes but has preferential affinity for the parallel four-stranded structure. In vitro radioimmunofilter binding experiments demonstrated that purified anti-DNA quadruplex antibodies from anti-quadruplex antibody-producing tissue culture supernatants have at least 10-fold higher affinity for quadruplexes than for triplex and duplex DNA structures of similar base composition and length. The antibody binds intramolecular DNA triplexes formed by d(G4T3G4T3C4) and d(C4T3G4T3G4), and the duplex d(CGCGCGCGCG)2 with an affinities of 6. 76 x 10(5), 5.59 x 10(5), and 8.26 x 10(5) M-1, respectively. Competition experiments showed that melted quadruplexes are not effective competitors for antibody binding when compared to native structures, confirming that the quadruplex is bound structure-specifically. To our knowledge, this is the first immunological reagent known to specifically recognize quadruplex structures. Subsequent sequence analysis demonstrates homologies between the antibody complementarity determining regions and sequences from Myb family telomere binding proteins, which are hypothesized to control cell aging via telomeric DNA interactions. The presence of this antibody in the autoimmune repertoire suggests a possible linkage between autoimmunity, telomeric DNA binding proteins, and aging.     

Spatial distribution and habitat characterization of schistosomiasis host snails in lake and land habitats of western Kenya

Intermediate host snails of schistosomiasis were surveyed in this study to determine their abundance and distribution in the lake and land aquatic habitats of Lake Victoria basin of Kenya. Several sites were sampled at eight locations, both in the lake and on the land. The habitat and/or vegetation type (i.e. open water, hippo grass, hyacinth, ambatch trees, other vegetation, stream, swamp, pond, dam) of the sampled aquatic sites within the locations were also differentiated, water physicochemical parameters were determined, and the abundance of different species or taxa of phytoplankton and zooplankton were enumerated and correlated with the abundance of schistosomiasis snails in the sites. The results indicated significantly more Biomphalaria sudanica snails than Bulinus africanus snails in different physical habitats on land (Student's t‐test, P < 0.05), as well as in different locations on land (Student's t‐test, P = 0.026). Regression analyses revealed that several physicochemical parameters, including dissolved oxygen (R² = ?0.659; n = 8; P = 0.014), pH (R² = 0.728; n = 8; P = 0.007) and turbulence (R² = ?0.616; n = 8; P = 0.02), were predictive of Biomphalaria spp. abundance, while pH (R² = 0.610; n = 8; P = 0.02) and turbulence (R² = ?0.578; n = 8; P = 0.028) were predictive of Bulinus spp. abundance in different locations in the lake. Cyanobacteria (R² = 0.638; n = 8; P = 0.02) and chlorophyceae (R² = ?0.50; n = 8; P = 0.05) were shown to be predictive of both Biomphalaria spp. and Bulinus spp. abundance in different locations in the lake. Zooplankton abundance varied significantly between different locations in the lake (One‐way anova , P < 0.001). Bosmina spp. were found to be predictive of both Biomphalaria spp. (R² = ?0.627; n = 8; P = 0.01) and Bulinus spp. (R² = ?0.50; n = 8; P = 0.05) in different locations in the lake. The results from this study will help inform policy regarding control measures for schistosomias and intermediate snail hosts in Lake Victoria waters, as well as in adjacent terrestrial aquatic habitats and even beyond.     

H] Tryptamine and 3H-water as diffusible internal standards for measuring brain extraction of radio-labeled substances following carotid injection

Urinary amino acid excretion was determined in 31 leukemic patients and 29 normal individuals by rapid gas chromatographic analysis of 16 amino acids as their N-acetyl-n-propyl esters. The leukemic patients were concurrently undergoing, or had recently completed, chemotherapy. It was found that aspartic acid, threonine, and serine were of significance in distinguishing between patients "on" therapy and those "off" therapy. Patients with advanced disease have the greatest aminoaciduria, although both the normal and leukemic populations have wide individual ranges. Within both populations, men excrete a greater variety and quantity of amino acids than women. It is concluded that analysis of urinary amino acids represents a history of complex metabolic events, which is potentially useful for evaluating patient response to chemotherapy in leukemia.     

Memory for Visuospatial Location Following Selective Hippocampal Sclerosis: The Use of Different Coordinate Systems.

This study addressed the role of the medial temporal lobe regions and, more specifically, the contribution of the human hippocampus in memory for body-centered (egocentric) and environment-centered (allocentric) spatial location. Twenty-one patients with unilateral atrophy of the hippocampus secondary to long-standing epilepsy (left, n = 7; right, n = 14) and 15 normal control participants underwent 3 tasks measuring recall of egocentric or allocentric spatial location. Patients with left hippocampal sclerosis were consistently impaired in the allocentric conditions of all 3 tasks but not in the egocentric conditions. Patients with right hippocampal sclerosis were impaired to a lesser extent and in only 2 of the 3 tasks. It was concluded that hippocampal structures are crucial for allocentric, but not egocentric, spatial memory. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Optimal greenhouse gas emissions in NGCC plants integrating life cycle assessment

The optimal design of an energy-intensive process involves a compromise between costs and greenhouse gas emissions, complicated by the interaction between optimal process emissions and supply chain emissions. We propose a method that combines generic abatement cost estimates and the results of existing (LCA) life cycle assessment studies, so that supply chain emissions are properly handled during optimization. This method is illustrated for a (NGCC) natural gas combined cycle power plant model with the following design and procurement options: procurement of natural gas from low-emissions producers, fuel substitution with (SNG) synthetic natural gas from wood, and variable-rate CO₂ capture and sequestration from both the NGCC and SNG plants. Using multi-objective optimization, we show two Pareto-optimal sets with and without the proposed LCA method. The latter can then be shown to misestimate CO₂ abatement costs by a few percent, penalizing alternate fuels and energy-efficient process configurations and leading to sub-optimal design decisions with potential net losses of the order of $1/MWh. Thus, the proposed LCA method can enhance the economic analysis of emissions abatement technologies and emissions legislation in general.     

Integrated manufacturing,employee and business performance: Australian and New Zealand evidence

This article reports on a large field study of Integrated Manufacturing (IM), consisting of Total Quality Management (TQM), Just in Time (JIT) and Advanced Manufacturing Technology (AMT). The relationship between these facets of Integrated Manufacturing and performance was examined empirically, including measures of employee performance and manufacturing performance. There was a significant correlation between employee performance and manufacturing performance. While all three facets of Integrated Manufacturing were relatively strongly connected with employee performance, manufacturing performance was significantly but less strongly connected to the intensity of TQM and JIT than employee performance, and not connected with the intensity of Advanced Manufacturing Technology. Some second-order interactive effects between the facets of Integrated Manufacturing and performance were found. Further, by partitioning the large data set into three using the AMT factor as a cutting variable, it was found that the stronger the AMT environment, the stronger the explanatory power of TQM and JIT on both employee and manufacturing performance. The interactive impact of AMT and TQM was significantly positively related to both employee and manufacturing performance.     

Quantitative detection method for Roundup Ready soybean in food using duplex real‐time PCR MGB chemistry

BACKGROUND: Methodologies that enable the detection of genetically modified organisms (GMOs) (authorized and non‐authorized) in food and feed strongly influence the potential for adequate updating and implementation of legislation together with labeling requirements. Quantitative polymerase chain reaction (qPCR) systems were designed to boost the sensitivity and specificity on the identification of GMOs in highly degraded DNA samples; however, such testing will become economically difficult to cope with due to increasing numbers of approved genetically modified (GM) lines. Multiplexing approaches are therefore in development to provide cost‐efficient solution. RESULTS: Construct‐specific primers and probe were developed for quantitative analysis of Roundup Ready ^® soybean (RRS) event glyphosate‐tolerant soybean (GTS) 40‐3‐2. The lectin gene (Le1) was used as a reference gene, and its specificity was verified. RRS‐ and Le1‐specific quantitative real‐time PCR (qRTPCR) were optimized in a duplex platform that has been validated with respect to limit of detection (LOD) and limit of quantification (LOQ), as well as accuracy. The analysis of model processed food samples showed that the degradation of DNA has no adverse or little effects on the performance of quantification assay. CONCLUSION: In this study, a duplex qRTPCR using TaqMan minor groove binder‐non‐fluorescent quencher (MGB‐NFQ) chemistry was developed for specific detection and quantification of RRS event GTS 40‐3‐2 that can be used for practical monitoring in processed food products. Copyright © 2010 Society of Chemical Industry     

rGO Sheets/ZnFe2O4 Nanocomposities as an Efficient Electro Catalyst Material for I3−/I− Reaction for High Performance DSSCs

In this paper, Reduced Graphene Oxide (rGO)/ZnFe₂O₄ (rZnF) nanocomposite is synthesized by a simple hydrothermal method and employed as a counter electrode (CE) material for tri-iodide redox reactions in Dye sensitized solar cells (DSSC) to replace the traditional high cost platinum (Pt) CE. X-ray diffraction analysis and High resolution Transmission electron microscopy, clearly indicated the formation of rZnF nanocomposite and also amorphous rGO sheets were smoothly distributed on the surface of ZnFe₂O₄ (ZnF) nanostructure. The rZnF-50 CE shows excellent electro catalytic activity toward I₃^? reduction, which has simultaneously been confirmed by cyclic voltammetry, electrochemical impedance spectroscopy and Tafel polarization measurements. A DSSC developed by rZnF-50 CE (η?=?8.71%) obtained quite higher than the Pt (η?=?8.53%) based CE under the same condition. The superior performances of rZnF-50 CE due to addition of graphene in to Spinel (ZnF) nanostructure results in creation of highly active electrochemical sites, fast electron transport linkage between CE and electrolyte. Thus it’s a promising low cost CE material for DSSCs.