Identification of molecular markers for aluminium tolerance in diploid oat through comparative mapping and QTL analysis.

The degree of aluminium tolerance varies widely across cereal species, with oats (Avena spp.) being among the most tolerant. The objective of this study was to identify molecular markers linked to aluminium tolerance in the diploid oat A. strigosa. Restriction fragment length polymorphism markers were tested in regions where comparative mapping indicated the potential for orthologous quantitative trait loci (QTL) for aluminium tolerance in other grass species. Amplified fragment length polymorphism (AFLP) and sequence-characterized amplified region (SCAR) markers were used to provide additional coverage of the genome. Four QTL were identified. The largest QTL explained 39% of the variation and is possibly orthologous to the major gene found in the Triticeae as well as Alm1 in maize and a minor gene in rice. A second QTL may be orthologous to the Alm2 gene in maize. Two other QTL were associated with anonymous markers. Together, these QTL accounted for 55% of the variation. A SCAR marker linked to the major QTL identified in this study could be used to introgress the aluminium tolerance trait from A. strigosa into cultivated oat germplasm.

Improving diagnostic yield of biliary brushings cytology for pancreatic cancer and cholangiocarcinoma.

Biliary brushings are currently the best accepted method to obtain a cytological diagnosis of pancreatic cancer or cholangiocarcinoma. The technique has good specificity but poor sensitivity. Two dedicated pathologists reviewed 137 consecutive biliary brushings from 127 patients between February 1997 and February 2000. The ultimate diagnosis was determined by review of radiology, operative diagnosis and patient outcome. The sensitivity, specificity, positive predictive value and negative predictive value of the original results and the review results were calculated and compared. Additional diagnostic categories 'suspicious' and 'atypical possibly benign' were included on review. After review, the sensitivity improved from 49.4% to 89.0% and the specificity remained 100%. The use of the additional diagnostic category 'suspicious' increased the sensitivity to 90.4%, at the expense of a fall of the specificity to 66.7%. We conclude that review by two dedicated pathologists and additional diagnostic categories can improve the diagnostic accuracy of biliary brushings.

A molecular linkage map with associated QTLs from a hulless x covered spring oat population.

In spring-type oat ( Avena sativa L.), quantitative trait loci (QTLs) detected in adapted populations may have the greatest potential for improving germplasm via marker-assisted selection. An F(6) recombinant inbred (RI) population was developed from a cross between two Canadian spring oat varieties: 'Terra', a hulless line, and 'Marion', an elite covered-seeded line. A molecular linkage map was generated using 430 AFLP, RFLP, RAPD, SCAR, and phenotypic markers scored on 101 RI lines. This map was refined by selecting a robust set of 124 framework markers that mapped to 35 linkage groups and contained 35 unlinked loci. One hundred one lines grown in up to 13 field environments in Canada and the United States between 1992 and 1997 were evaluated for 16 agronomic, kernel, and chemical composition traits. QTLs were localized using three detection methods with an experiment-wide error rate of approximately 0.05 for each trait. In total, 34 main-effect QTLs affecting the following traits were identified: heading date, plant height, lodging, visual score, grain yield, kernel weight, milling yield, test weight, thin and plump kernels, groat beta-glucan concentration, oil concentration, and protein. Several of these correspond to QTLs in homologous or homoeologous regions reported in other oat QTL studies. Twenty-four QTL-by-environment interactions and three epistatic interactions were also detected. The locus controlling the covered/hulless character ( N1) affected most of the traits measured in this study. Additive QTL models with N1 as a covariate were superior to models based on separate covered and hulless sub-populations. This approach is recommended for other populations segregating for major genes. Marker-trait associations identified in this study have considerable potential for use in marker-assisted selection strategies to improve traits within spring oat breeding programs.

Infrared and EPR Spectroscopic Studies of 2-C(2)H(2)F and 1-C(2)H(2)F Radicals Isolated in Solid Argon.

2-fluorovinyl radicals were generated in solid argon by solid-state chemical reactions of mobile F atoms with acetylene and its deuterated analogues. Highly resolved EPR spectra of the stabilized radicals CHF&dbond;(*)CH, CDF&dbond;(*)CD, CHF&dbond;(*)CD, and CDF&dbond;(*)CH were obtained for the first time. The observed spectra were assigned to cis-2-fluorovinyl radical based on excellent agreement between the measured (a(F) = 6.50, a(betaH) = 3.86, a(alphaH) = 0.25 mT) hyperfine constants and those calculated using density functional (B3LYP) theory. Analogous experiments carried out using infrared spectroscopy yielded a complete assignment of the vibrational frequencies. An unusual reversible photochemical conversion is observed in which cis-2-fluorovinyl radicals can be partially converted to 1-fluorovinyl radicals by pulsed laser photolysis at 532 nm. Photolysis at 355 nm converts 1-fluorovinyl back to cis-2-fluorovinyl. High-resolution EPR and infrared spectra of 1-fluorovinyl were obtained for the first time. The measured hyperfine constants (a(F) = 13.71, a(H1) = 4.21, a(H2) = 1.16 mT) are in good agreement with calculated values. Copyright 2001 Academic Press.

Donor Action: an international initiative to alleviate organ shortage.

CONTEXT: Donor Action, an international initiative to alleviate organ shortage, provides a comprehensive state-of-the-art methodology that helps critical care units develop a tailor-made approach to optimize donation practices and performance. OBJECTIVE: To report the impact of the Donor Action methodology on organ donation rates in 8 countries (70 critical care units) in North America and Europe. DESIGN: Baseline data on the clinical potential for donation, staff attitudes, knowledge toward donation, self-reported confidence in performing a range of donation roles, and educational requirements were gathered. These data were analyzed using the Donor Action database and improvement measures were introduced to address identified weaknesses. RESULTS: Following introduction of the program's improvement measures, which addressed identified weaknesses, donations increased on average by 53% (P = .0017) per country at 1 year. Sustained improvements settled at 70% to 160% increases at 3 years. Although Donor Action is at various stages of implementation in different countries, the number of centers and countries demonstrating an immediate awareness effect is increasing and sustained effects in centers with the longest follow-up promise a significant impact on donation rates as more countries implement this methodology.

A European perspective on organ procurement: breaking down the barriers to organ donation.

A worldwide shortage of donor organs has led to the development of national and international systems for organ procurement and allocation. Such systems promote organ donation and ensure fair distribution of available donor organs through a combination of legislation, organ exchange organizations (OEOs), transplant coordinators, publicity campaigns, donor cards, and professional training programs. The development of national and international OEOs is central to this process because they maintain waiting lists and allocate organs in the most appropriate way. Most countries also employ transplant coordinators whose role involves promoting links between transplant centers and intensive care units, establishing protocols for organ donation, and helping hospital staff deal with the sensitive issues involved in organ donation. Educational initiatives, such as the European Donor Hospital Education Programme developed by Eurotransplant is now used in over 30 countries worldwide. The program aims to improve professionals' understanding of the legal and ethical issues involved in transplantation, to help them communicate effectively and sympathetically with bereaved families, and to increase organ donation rates. Other initiatives include programs such as the Donor Action Programme, which was set up by professional organizations in the US and Europe aiming to help hospitals establish tailor-made organ procurement policies to ensure that all potential donors can be identified and reported and the needs of unfortunate families can be met in a caring and sensitive manner.

The European Donor Hospital Education Programme (EDHEP): addressing the training needs of doctors and nurses who break bad news, care for the bereaved, and request donation.

The competence of critical care staff when it comes to death and organ donation can make the difference between a family's agreeing to or refusing the latter. Doctors and nurses often feel uncomfortable approaching relatives about donation and attribute this to a lack of training. Bereaved relatives express dissatisfaction with inappropriate communication and support when brain death is announced and thereafter when a request for donation is made. The European Donor Hospital Education Programme (EDHEP) was designed to meet the training needs of critical care staff in breaking bad news, caring for the bereaved, and requesting donation. EDHEP is a two-part educational package consisting of a presentation about the donor shortage followed by a one-day workshop. The implementation of EDHEP throughout the world has been facilitated through effective national working groups and standardised "train the trainer" courses. Several countries anecdotally report increases in donation following implementation. Controlled evaluation of the effect(s) of EDHEP, which started at the end of 1995, focuses on the satisfaction of the participants with EDHEP, on the competence of the participants in breaking bad news and requesting donation, on the teamwork regarding death and donation, on the satisfaction of bereaved relatives, and on organ donation rates.

Hospital attitudes: preliminary findings from donor action pilot projects.

PURPOSE: As part of the Donor Action collaboration (Eurotransplant Foundation, The Netherlands; Organización National de Transplantes, Spain; and The Partnership for Organ Donation, USA), a hospital survey was administered to gather baseline data on staff attitudes about organ donation and level of self-reported skills/confidence in performing a range of organ donation roles. METHODS: A standard survey instrument was administered in two hospitals in Spain, two in The Netherlands and one in the UK. In four hospitals the survey was administered to all ICU staff; in one hospital it was administered to a random sample of hospital staff. The instrument was created in English, and translated into Spanish and Dutch for the hospitals in each country. RESULTS: Data were analysed by country and showed consistently strong perceptions that organ donation saves lives (97%). Support for donation (95%) and willingness to donate one's own organs (82%) were high in all three country samples. Significant differences in belief were observed when respondents were asked whether they agreed that organ donation helps families with their grief. The UK respondents were more likely to agree (57%), with lower levels of agreement in Spain (47%) and The Netherlands (14%) (P < 0.0001). Average ratings of skills/confidence were highest for notifying the transplant coordinator (49%) and comforting the family (48%) with lower confidence reported regarding explaining brain death (34%), introducing organ donation (32%), and requesting organ donation (26%). Ratings varied widely across countries with UK respondents expressing the highest level of self-reported confidence, and Spanish respondents the lowest level. For example, 77% of UK respondents reported themselves skilled/confident explaining brain death, versus 47% of Dutch respondents and 11% of Spanish respondents (P < 0.0001). Similar results were seen regarding requesting organ donation: UK 53%; The Netherlands 30%; Spain 13% (P < 0.0001). CONCLUSIONS: There has been a lack of data about hospital staff attitudes and skills to allow for comparison across national systems, and to support the targeting of specific strategies to the needs within different countries. These results show the feasibility of collecting and comparing data across national systems. These pilot findings also suggest that there may be important differences in attitudes and self-perceived skills/confidence across countries. Work remains to correlate attitudes and self-perceived skills to actual performance. It is noteworthy that the sense of staff preparedness was lowest in Spain which has the highest donation rates. This may reflect the degree to which role specialization in donation has been successfully integrated into hospital practice. Expansion of the survey to additional hospitals will help to answer such questions.

Kinetics in solids.

The kinetics of solid state reactions generally cannot be assumed to follow simple rate laws that are applicable to gas-phase reactions. Nevertheless, a widely practiced method for extracting Arrhenius parameters from thermal analysis experiments involves force fitting of experimental data to simple reaction-order kinetic models. This method can produce significant errors in predicted rates outside the experimental range of temperatures, and it is of limited utility for drawing mechanistic conclusions about reactions. In this review, we discuss how an alternative "model-free" approach to kinetic analysis, which is based on the isoconversional method, can overcome some of these limitations.

Organ shortages: maximising the donor potential.

Of all the problems foreseen in the pioneering days of organ transplantation, a shortage of donor organs, was not even remotely considered as a barrier to progress. Such has been the success of transplantation over the last two decades, organ shortage is now considered the major limitation. This chapter will concentrate on efforts to increase the donor potential. The development of Organ Exchange Organizations is briefly described with special emphasis on their role in organ allocation systems to avoid wastage of this precious resource, procurement transplant coordinators, the organization of organ procurement and the consent process. We look briefly at the influence of the media and end with some considerations on how to maximise the current supply of organs for transplantation.

Gastric outflow obstruction caused by gall stones and leading to death by complex metabolic derangement.

A 67 year old woman was admitted with a three week history of vomiting, having become increasingly confused for three days. Investigations revealed deranged serum biochemistry consistent with a combination of a diabetic non-ketotic hyperosmolar state and a metabolic alkalosis consistent with gastric outflow obstruction. She was treated with intravenous saline, intravenous insulin, and subcutaneous heparin, but did not improve clinically and had an asystolic cardiac arrest the following day; she was transferred to the intensive care unit and despite treatment with inotropes she died 40 hours after admission. Necropsy revealed that the stomach was massively dilated with gas and stomach contents, and contained many small black faceted gall stones. In addition a large nonfaceted brown-yellow gall stone was wedged in the pyloric antrum causing total obstruction. The patient had died from a complex metabolic derangement including non-ketotic hyperosmotic diabetic coma and metabolic alkalosis precipitated by the acute gastric outflow obstruction complicated by previously undiagnosed type II diabetes mellitus.

The identification of random amplified polymorphic DNA markers for daylength insensitivity in oat.

Daylength insensitive accessions of Avena sativa L. are being used to develop cultivars that will flower normally when grown under short or long photoperiods. Field data indicate that the insensitivity trait is under the control of a single dominant gene, designated Di1. The random amplified polymorphic DNA (RAPD) technique and bulk segregant analysis of daylength sensitive and insensitive plants were used to find markers for this gene. Five of 200 random decamer primers tested produced polymorphic bands, which were shown to be linked to the trait using 30 homozygous insensitive and 30 homozygous sensitive F3 individuals. Three of the markers produced a band in the presence of the dominant allele, and two in its absence. Segregation analysis showed that markers 221 and 136 could be mapped to within 9.8 +/- 4.6 and 13.9 +/- 5.4 cM of the trait, respectively; that is, close enough to be useful in a breeding program. A study of different cultivars suggested that the band produced by primer 136 is actually the more closely linked marker and the only one present in the original Di1 gene donor CAV2700. The possibility of using both markers in populations derived from different cultivars is discussed.