Synthesis and application of circularizable ligation probes.

We describe a PCR-based approach for the synthesis of circularizable ligation probes (CLiPs). CLiPs are single-stranded probes that consist of target-specific ends separated by a noncomplementary "linker" sequence. When hybridized to a target, the CLiP forms a nicked circle that may be sealed by DNA ligase only if the 5' and 3' ends show perfect Watson-Crick base pairing, thus enabling the discrimination of single nucleotide polymorphisms. Primers incorporating target sequence at their 5' end and plasmid sequence at the 3' end were used in a PCR amplification. In addition, the antisense primer was 5' labeled with biotin, and the amplification was performed in the presence of fluorescently labeled dUTP. The resulting PCR product was captured with streptavidin-coated paramagnetic beads, and the top strand, which forms the CLiP, was alkali eluted. This PCR-based method has allowed the synthesis of CLiPs that are larger and more highly labeled than has previously been possible, with ligation efficiencies similar to those of the purest chemically synthesized padlock probes. Ligations performed in the presence of cognate or mismatched sequence were analyzed by denaturing PAGE using a fluorescent DNA sequencer. Genotyping using target immobilized to nylon membranes was also performed. The CLiPs were readily able to distinguish between mutant and wild-type alleles for the common genetic disorder, 21-hydroxylase deficiency. Additionally, CLiPs of different lengths were synthesized and compared.

Antiphospholipid antibody syndrome: implications for nursing practice.

Lupus anticoagulant and anticardiolipin are antiphospholipid antibodies. The presence of either or both antiphospholipid antibodies may result in antiphospholipid antibody syndrome, which is typified by recurrent venous or arterial thrombosis, recurrent fetal loss, and thrombocytopenia. Many features of this syndrome are not well understood, including its prevalence, how and why antiphospholipid autoantibodies develop, and their definitive role in disease states. In clinical practice, nurses care for patients and families with antiphospholipid antibody syndrome who require treatment, education, and support.

A cross-over study of the effect of a single oral feeding of medium chain triglyceride oil vs. canola oil on post-ingestion plasma triglyceride levels in healthy men.

Due to its unique absorption and metabolism characteristics, medium chain triglyceride (MCT) oil, consisting of fatty acids with 8-12 carbons, has been used therapeutically since the 1950s in the treatment of fat malabsorption, cystic fibrosis, epilepsy, weight control, and to increase exercise performance. Medium chain triglycerides are easily hydrolyzed in the intestines and the fatty acids are transported directly to the liver via the portal venous system, in contrast to long-chain fatty acids (LCFAs), which are incorporated into chylomicrons for transport through the lymphatic system or peripheral circulation. Medium chain fatty acids (MCFAs) do not require carnitine to cross the double mitochondrial membrane of the hepatocyte, thus they quickly enter the mitochondria and undergo rapid beta-oxidation, whereas most LCFAs are packaged into triglycerides in the hepatocyte. In this single-blind, randomized, cross-over study, 20 healthy men ingested a single dose of either 71 g of MCT oil or canola oil. Blood samples were taken at baseline and at hours one through five post-ingestion to compare the effect of a single oral dosing of MCT oil versus canola oil on post-ingestion plasma triglyceride levels. Mean triglyceride values after canola oil increased 47 percent above baseline (p <0.001), while mean triglyceride values after MCT oil decreased 15 percent from baseline (p <0.001), which is consistent with several other studies involving short- and longer-term feeding with MCT oil. The effect of long-term usage of MCT oil on triglycerides is yet to be established.

Outcomes-based education in a critical care nursing course.

Educational institutions are being asked to develop more meaningful ways to measure student learning. The critical care nursing faculty at this baccalaureate institution have delineated a process of outcomes-based education that they use as evidence of successful learning in their course. The faculty regard this as an ongoing project in which this evidence can only lead to continuing improvement.

Case studies: what a difference a nurse makes.

The case studies represent three different types of critical care nursing challenges. The objective of each case study is to identify a key nursing problem and focus on nursing research or nursing behavior that made or could have made the greatest difference to the patient outcome. The first case is of a man with Buerger's disease who had unmet needs regarding pain control. The second patient had an out-of-hospital ruptured abdominal aortic aneurysm and survived a rough, 7-week hospitalization. Major complications included coagulopathy and rhabdomyolysis. Critical care nursing expertise in early assessment and intervention was integral to survival for this patient. The third case is of an elderly woman with an extensive medical history who experienced repeated prosthetic aortic graft infections, skin breakdown, and poor wound healing. The highlighted research is on low air loss beds. The critical care nurse made a significant difference in increasing each of these patient's chances for a successful hospital outcome.

A comparison of learner outcomes to traditional and videotaped classes.

A course was taught to registered nurse students using videotaped classes shown on a bus while students commuted to and from a distant clinical site. Course outcomes for this group were compared with course outcomes for a traditional classroom-taught group of registered nurse students. Outcomes were quite similar, thus making this method of technology use and time management an appropriate teaching outreach/recruitment technique for the college to use with the registered nurse student population.

Multisystem complications of bulimia: a critical care case.

Bulimia is an eating disorder characterized by behaviors which can lead to very serious health problems. Behaviors such as binging and purging and abuse of emetics, diuretics, or laxatives can cause serious complications, sometimes of a critical nature. Nurses must maintain a high index of suspicion for this disorder and use skillful assessment and intervention to prevent the critical multisystem complications of bulimia.

Critical care leadership course: a recruitment and retention strategy.

This article describes how to implement a baccalaureate level course in critical care leadership and management and emphasizes the value of this course in recruiting and developing critical care nurses with leadership abilities.

Selection and characterization of mutants of Phanerochaete chrysosporium exhibiting ligninolytic activity under nutrient-rich conditions.

Synthesis of the ligninolytic system of the wood-degrading fungus Phanerochaete chrysosporium is induced during secondary metabolism, brought about by nitrogen, carbon, or sulfur starvation. We describe here a strategy for selection of mutants which are ligninolytic (lignin----CO2) and overproduce lignin-degrading enzymes (ligninases) under nutrient-rich conditions (during primary metabolism). The strategy is based on using an adduct of lysine and a lignin model compound. Ligninase-dependent oxidation of this adduct releases free lysine, which complements the lysine requirements of a lysine auxotroph. Accordingly, a lysine auxotroph was mutagenized by UV irradiation and survivors were plated onto medium containing the adduct and high ammonia nitrogen. Four mutants which overproduce the ligninase isozymes were isolated by this procedure. Further characterization of one of the mutants, PSBL-1, indicated that the predominant isozymes produced are H1 (pI = 4.7) and H2 (pI = 4.4). The ligninase activity of PSBL-1, measured by veratryl alcohol oxidation, peaks on day 5 at over 1,000 U.liter-1. The mutant PSBL-1 was also able to degrade [14C]lignin to 14CO2, indicating that the complete ligninolytic system is deregulated.

Overview of inflammatory bowel disease.

Ulcerative colitis and Crohn's disease have been on the increase in terms of incidence, prevalence, and virulence. The young adult population is affected most frequently and most severely. The cause of inflammatory bowel disease is unknown, but many misconceptions exist about its etiology. Nurses need to be familiar with the similarities and differences between the two illnesses, the treatment, and the prognosis in order to be effective care givers. Good health care can make a significant contribution to the quality of life the patient subsequently has.