Optimizing Reaction Conditions of the NanoOrange® Protein Quantitation Method for Use With Microplate-based Automation

Because of the rapidly expanding need for higher sample throughput in drug discovery, automation of corresponding biochemical analyses is desirable. In particular, automation of protein quantitation is crucial since its results are used extensively. Recently, a single-reagent fluorescent protein quantitation method (NanoOrange®) with attractive performance attributes has become available. While it can potentially be automated with liquid handling workstations, several of this method's reaction parameters need to be optimized.

We studied the time and a temperature dependence of the NanoOrange protein quantitation reaction in ninety-six well black microplates using either a temperature-regulated hot block or a microwave oven as heat sources. Fluorescence of the NanoOrange reaction was quantified with a multimode microplate spectral scanner.

Time-dependent heating profiles of filled microplates placed on hot blocks at fixed temperatures (45, 55, 65, 75, and 95°C) revealed temperature differences of 4–7°C cooler for the outside wells compared to the inner wells, however the maximum well temperature did not exceed 65°C. Similar time-temperature studies of microwave-heated microplates revealed an equilibrium temperature of 45–49°C that was 10–16°C lower than microplates that were block heated.

The bovine serum albumin (BSA): NanoOrange standard curves created using a hot block increased in slope from 45°C to 55°C, but then remained constant from 65 to 85°C.

Fluorescence of the BSA: NanoOrange standard curve created using a microwave oven was about half the magnitude of the hot block-derived curves, possible reflecting a lower energy transfer rate of the microwave oven. We conclude that the NanoOrange protein quantitation method can be automated if a microplatecompatible hot block with a 65-85°C surface can heat the microplate for minimum of 15 min prior to quantifying the reaction's fluorescence.     

Transport and related properties of paint films. I. Composition and structure

This is the first of a series of articles on ionic and molecular transport processes and related properties of paint films. The intention is to correlate adhesion and initiation of corrosion to these relevant physical and chemical properties. Chemical formulas and structure of three vinyl–three epoxy– and one polyurethane–paint system are given in this article. Each paint has been formulated to meet appropriate military specifications. Subsequent articles will deal with dynamic mechanical properties, small angle x-ray scattering measurements of porosity, and transport properties.     

Review of Recent developments in psychoanalysis.

Reviews the book, Recent developments in psychoanalysis by Morris N. Eagle (see record 1987-98252-000). This volume is a timely and important addition to psychoanalytic literature. Eagle states that it is "intended as a systematic overview of recent ideas with the aim of providing a coherent account of the current status of psychoanalytic theory." His overview is generally fair, but penetrating, and exposes much of the irrationality and confusion that have plagued psychoanalytic thinking recently. He suggests that this confusion has arisen out of the tendency of psychoanalytic theorists to ignore advances in other areas of psychological research and related fields, particularly in the area of psychology of child development. Further confusion has been created by their attempt to formulate new concepts and ideas within the framework of classical Freudian metapsychology, which is rooted in instinct theory. These attempts, in many cases, have led to the use of a psychoanalytic language full of jargon, rendering theoretical formulations expressed in this way totally incomprehensible and aloof from logical consideration, while pretending to add metapsychological depth. Ultimately, however, he tries to elucidate the many real contributions to psychoanalytic theory by these theorists and to integrate their work into a coherent framework. His efforts in this regard reflect a sympathetic approach to the aims and ideals of psychoanalysis as a social science and a form of therapy. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

The Claims of Multiculturalism and Journalism's Promise of Diversity

Mainstream American journalism rests on a pluralist model of democracy that conserves the status quo, essentializes culture, and trivializes diversity. A very different understanding of diversity emerges from a multiculturalist perspective that questions existing arrangements, posits a relational view of culture, and defines diversity in terms of patterns of discrimination and inequality. A case study of coverage of a local issue in the mainstream and minority press underpins a discussion of the importance of diversifying journalism by restructuring the American press. Journalism diversity matters most not only as it heightens sensitivity to cultural differences but as it strengthens the role of minority media in the struggle to achieve the social justice and political parity that a culturally diverse society demands.     

Thermochemical modeling of refractory corrosion in slagging coal gasifiers

Slagging coal gasifiers suffer corrosive attack on the refractory liner and these interactions were thermochemically simulated. The slag is observed to penetrate the refractory, which complicates modeling the phase behavior of the slag-penetrated interior of the refractory. A simple strategy was adopted such that stepwise changes in composition with decreasing slag content were assumed to account for the compositional changes as slag penetrates the refractory. The thermochemical equilibrium calculations following this strategy typically yielded three solution phases as well as the stoichiometric crystalline phases AlPO₄ and Ca₃(PO₄)₂ depending on composition/penetration. Under some conditions a slag liquid miscibility gap exists such that two slag liquids coexist.     

Structural characterization of Ti and Pt thin films on GaAs(100) substrate

In an attempt to understand the Schottky barrier behavior of Ti/Pt/GaAs and Pt/Ti/ GaAs bimetal Schottky diodes, we have investigated the interfacial morphology of Ti and Pt thin films on GaAs(l00) substrate. The characterization was based on coverage profiling of Auger electron spectroscopy in conjunction with transmission electron microscopy. Emphasis was placed on film uniformity and atomic interdiffusion. The results showed Ga and As outdiffusion in Pt/GaAs interface and some oxygen incorporated in Ti film, but no evidence of clustering for both metal/GaAs systems.     

Properties of single-walled carbon nanotube-based aerogels as a function of nanotube loading

Here, we present the synthesis and characterization of low-density single-walled carbon nanotube-based aerogels (SWNT-CA). Aerogels with varying nanotube loading (0–55 wt.%) and density (20–350 mg cm^?3) were fabricated and characterized by four-probe method, electron microscopy, Raman spectroscopy and nitrogen porosimetry. Several properties of the SWNT-CAs were highly dependent upon nanotube loading. At nanotube loadings of 55 wt.%, shrinkage of the aerogel monoliths during carbonization and drying was almost completely eliminated. Electrical conductivities are improved by an order of magnitude for the SWNT-CA (55 wt.% nanotubes) compared to those of foams without nanotubes. Surface areas as high as 184 m² g^?1 were achieved for SWNT-CAs with greater than 20 wt.% nanotube loading.     

Relations between incompatible displacement model and hybrid stress model

An examination of the variational formulations confirms the similarity between the incompatible displacement model and the assumed stress hybrid model that was pointed out by Irons in 1972. But the basic differences between the two are also identified. For 8-node solid elements the assumed stress terms obtained through a rational procedure also agree with those deduced by Irons purely from his physical insight.     

The Effects of Omega‐3 Polyunsaturated Fatty Acid Consumption on Mammary Carcinogenesis

The consumption of omega‐3 polyunsaturated fatty acids (n‐3 PUFA) is associated with a reduced risk of breast cancer. Studies in animals and in vitro have demonstrated mechanisms that could explain this apparent effect, but clinical and epidemiological studies have returned conflicting results on the practical benefits of dietary n‐3 PUFA for prevention of breast cancer. Effects are often only significant within a population when comparing the highest n‐3 PUFA consumption group to the lowest n‐3 group or highest n‐6 group. The beneficial effects of n‐3 PUFA eicosapentaenoic and docosahexaenoic on the risk of breast cancer are dose dependent and are negatively affected by total n‐6 consumption. The majority of the world population, including the most highly developed regions, consumes insufficient n‐3 PUFA to significantly reduce breast cancer risk. This review discusses the physiological and dietary context in which reduction of breast cancer risk may occur, some proposed mechanisms of action and meaningful recommendations for consumption of n‐3 PUFA in the diet of developed regions.     

Persistent organic pollutants in fish tissue in the mid-continental great rivers of the United States

Great rivers of the central United States (Upper Mississippi, Missouri, and Ohio rivers) are valuable economic and cultural resources, yet until recently their ecological condition has not been well quantified. In 2004-2005, as part of the Environmental Monitoring and Assessment Program for Great River Ecosystems (EMAP-GRE), we measured legacy organochlorines (OCs) (pesticides and polychlorinated biphenyls, PCBs) and emerging compounds (polybrominated diphenyl ethers, PBDEs) in whole fish to estimate human and wildlife exposure risks from fish consumption. PCBs, PBDEs, chlordane, dieldrin and dichlorodiphenyltrichloroethane (DDT) were detected in most samples across all rivers, and hexachlorobenzene was detected in most Ohio River samples. Concentrations were highest in the Ohio River, followed by the Mississippi and Missouri Rivers, respectively. Dieldrin and PCBs posed the greatest risk to humans. Their concentrations exceeded human screening values for cancer risk in 27-54% and 16-98% of river km, respectively. Chlordane exceeded wildlife risk values for kingfisher in 11-96% of river km. PBDE concentrations were highest in large fish in the Missouri and Ohio Rivers (mean > 1000 ng g^− 1 lipid), with congener 47 most prevalent. OC and PBDE concentrations were positively related to fish size, lipid content, trophic guild, and proximity to urban areas. Contamination of fishes by OCs is widespread among great rivers, although exposure risks appear to be more localized and limited in scope. As an indicator of ecological condition, fish tissue contamination contributes to the overall assessment of great river ecosystems in the U.S.