Application of two-dimensional gel electrophoresis to interrogate alterations in the proteome of genetically modified crops. 1. Assessing analytical validation

Current tools used to assess the safety of food and feed derived from modern biotechnology emphasize the investigation of possible unintended effects caused directly by the expression of transgenes or indirectly by pleiotropy. These tools include extensive multisite and multiyear agronomic evaluations, compositional analyses, animal nutrition, and classical toxicology evaluations. Because analytical technologies are rapidly developing, proteome analysis based on two-dimensional gel electrophoresis (2DE) was investigated as a complementary tool to the existing technologies. A 2DE method was established for the qualitative and quantitative analysis of the seed proteome of Arabidopsis thaliana with the following validation parameters examined: (1) source and scope of variation; (2) repeatability; (3) sensitivity; and (4) linearity of the method. The 2DE method resolves proteins with isoelectric points between 4 and 9 and molecular masses (MM) of 6-120 kDa and is sensitive enough to detect protein levels in the low nanogram range. The separation of the proteins was demonstrated to be very reliable with relative position variations of 1.7 and 1.1% for the pI and MM directions, respectively. The mean coefficient of variation of 254 matched spot qualities was found to be 24.8% for the gel-to-gel and 26% for the overall variability. A linear relationship (R2 > 0.9) between protein amount and spot volume was demonstrated over a 100-fold range for the majority of selected proteins. Therefore, this method could be used to interrogate proteome alterations such as a novel protein, fusion protein, or any other change that affects molecular mass, isoelectric point, and/or quantity of a protein.     

Massive-star supernovae as major dust factories

We present late-time optical and mid-infrared observations of the Type II supernova 2003gd in the galaxy NGC 628. Mid-infrared excesses consistent with cooling dust in the ejecta are observed 499 to 678 days after outburst and are accompanied by increasing optical extinction and growing asymmetries in the emission-line profiles. Radiative-transfer models show that up to 0.02 solar masses of dust has formed within the ejecta, beginning as early as 250 days after outburst. These observations show that dust formation in supernova ejecta can be efficient and that massive-star supernovae could have been major dust producers throughout the history of the universe.     

A cortical region consisting entirely of face-selective cells

Face perception is a skill crucial to primates. In both humans and macaque monkeys, functional magnetic resonance imaging (fMRI) reveals a system of cortical regions that show increased blood flow when the subject views images of faces, compared with images of objects. However, the stimulus selectivity of single neurons within these fMRI-identified regions has not been studied. We used fMRI to identify and target the largest face-selective region in two macaques for single-unit recording. Almost all (97%) of the visually responsive neurons in this region were strongly face selective, indicating that a dedicated cortical area exists to support face processing in the macaque.     

Distribution of CCR3 mRNA expression in horse tissues

CCL11 (also known as eotaxin) is a very potent and selective mediator of eosinophil migration which exerts its effects through its receptor, CCR3. In this study we report the cloning of an equine CCR3 cDNA sequence and investigation of the localization of CCR3 mRNA expression in horse tissues. Equine CCR3 displayed high levels of sequence identity with CCR3 sequences in other species. RT-PCR analysis revealed the expression of CCR3 in colon, lung and spleen of normal horses. In situ hybridisation experiments indicated that expression of CCR3 mRNA in colon was predominantly in eosinophils and to a lesser extent in mast cells, whereas CCR3 was seen mainly in lymphocytes of the lung and spleen. In view of the role of CCR3 in the recruitment of cells into sites of allergic inflammation, equine-specific CCR3 sequence data and information on tissue localization will be of potential benefit in the development of CCR3-targeted anti-inflammatory therapies in the horse.     

National diet and nutrition surveys: the British experience

OBJECTIVE: The National Diet and Nutrition Surveys (NDNS) are a series of government-funded surveys of food intake, nutrient intake and nutritional status of individuals, undertaken to support nutritional policy and risk assessment. This paper summarises a review that considered the extent to which NDNS met the needs of users and suggested options for the future. The Food Standards Agency has since progressed favoured options. This paper aims to help others wishing to obtain this type of information within their own populations. DESIGN: A detailed questionnaire was used to probe use of data and gather opinions from users, producers and managers of the NDNS. It asked about general information needs from NDNS and changes that might be made. This was followed by a two-day workshop which included discussion of the main issues and the generation of 19 possible future options for consideration by the Agency. RESULTS: Options to improve effectiveness included methods to prioritise breadth and depth of coverage and possible ways of improving response and compliance. Strategies to make surveys more efficient and timely, such as adopting a rolling programme, disaggregating survey components, integrating with other studies and improving data access, were also suggested. A rolling programme, in which data are collected continuously, was the favoured option to address some of the concerns and a strategy is now in place to achieve this. CONCLUSIONS: There is widespread support for the NDNS from its users. There is no alternative source for such high-quality data on food and nutrient consumption and nutritional status and physical measurements in the same individuals. Useful information, such as the potential value of using a rolling programme from the outset, can be gained from this British experience by others wishing to measure food and nutrient intakes and status in their own populations.     

Long-term persistence of multi-drug-resistant Salmonella enterica serovar Newport in two dairy herds

OBJECTIVE: To evaluate the association between maintaining joint hospital and maternity pens and persistence of multi-drug-resistant (MDR) Salmonella enterica serovar Newport on 2 dairy farms. DESIGN: Observational study. SAMPLE POPULATION: Feces and environmental samples from 2 dairy herds. PROCEDURE: Herds were monitored for fecal shedding of S enterica Newport after outbreaks of clinical disease. Fecal and environmental samples were collected approximately monthly from pens housing sick cows and calving cows and from pens containing lactating cows. Cattle shedding the organism were tested serially on subsequent visits to determine carrier status. One farm was resampled after initiation of interventional procedures, including separation of hospital and maternity pens. Isolates were characterized via serotyping, determination of antimicrobial resistance phenotype, detection of the CMY-2 gene, and DNA fingerprinting. RESULTS: The prevalence (32.4% and 33.3% on farms A and B, respectively) of isolating Salmonella from samples from joint hospital-maternity pens was significantly higher than the prevalence in samples from pens housing preparturient cows (0.8%, both farms) and postparturient cows on Farm B (8.8%). Multi-drug-resistant Salmonella Newport was isolated in high numbers from bedding material, feed refusals, lagoon slurry, and milk filters. One cow excreted the organism for 190 days. Interventional procedures yielded significant reductions in the prevalences of isolating the organism from fecal and environmental samples. Most isolates were of the C2 serogroup and were resistant to third-generation cephalosporins. CONCLUSIONS AND CLINICAL RELEVANCE: Management practices may be effective at reducing the persistence of MDR Salmonella spp in dairy herds, thus mitigating animal and public health risk.     

Evaluation of strontium 90 irradiation in treatment of cutaneous mast cell tumors in cats: 35 cases (1992-2002)

OBJECTIVE: To determine the efficacy of strontium 90 beta irradiation in the management of cutaneous mast cell tumors (CMCTs) in cats. STUDY DESIGN: Retrospective case series. ANIMALS: 35 client-owned cats with CMCTs. PROCEDURE: Medical records of cats with CMCTs in which tumors were radiated by use of a strontium 90 ophthalmic applicator from 1992 to 2002 were reviewed. Cats were included if CMCT was diagnosed, there were no other sites of MCT involvement at the time of treatment, and records contained adequate follow-up information to permit retrospective assessment of local tumor control. RESULTS: 54 tumors in 35 cats were treated with a median dose of 135 Gy of strontium 90 beta irradiation, resulting in local tumor control in 53 of 54 (98%) tumors with a median follow-up time of 783 days after treatment. Median survival time was 1,075 days. Adverse effects of treatment appeared to be infrequent and of mild severity. CONCLUSIONS AND CLINICAL RELEVANCE: Results indicated that strontium 90 beta irradiation resulted in long-term tumor control and should be considered an effective alternative to surgical resection in management of CMCTs in cats.     

The technology path to deep greenhouse gas emissions cuts by 2050: the pivotal role of electricity

Several states and countries have adopted targets for deep reductions in greenhouse gas emissions by 2050, but there has been little physically realistic modeling of the energy and economic transformations required. We analyzed the infrastructure and technology path required to meet California's goal of an 80% reduction below 1990 levels, using detailed modeling of infrastructure stocks, resource constraints, and electricity system operability. We found that technically feasible levels of energy efficiency and decarbonized energy supply alone are not sufficient; widespread electrification of transportation and other sectors is required. Decarbonized electricity would become the dominant form of energy supply, posing challenges and opportunities for economic growth and climate policy. This transformation demands technologies that are not yet commercialized, as well as coordination of investment, technology development, and infrastructure deployment.