Quantitative profiling and pattern analysis of triacylglycerol species in Arabidopsis seeds by electrospray ionization mass spectrometry.

Plant triacylglycerols (TAGs), or vegetable oils, provide approximately 25% of dietary calories to humans and are becoming an increasingly important source of renewable bioenergy and industrial feedstocks. TAGs are assembled by multiple enzymes in the endoplasmic reticulum from building blocks that include an invariable glycerol backbone and variable fatty acyl chains. It remains a challenge to elucidate the mechanism of synthesis of hundreds of different TAG species in planta. One reason is the lack of an efficient analytical approach quantifying individual molecular species. Here we report a rapid and quantitative TAG profiling approach for Arabidopsis seeds based on electrospray ionization tandem mass spectrometry with direct infusion and multiple neutral loss scans. The levels of 93 TAG molecular species, identified by their acyl components, were determined in Arabidopsis seeds. Quantitative TAG pattern analyses revealed that the TAG assembly machinery preferentially produces TAGs with one elongated fatty acid. The importance of the selectivity in oil synthesis was consistent with an observation that an Arabidopsis mutant overexpressing a patatin-like phospholipase had enhanced seed oil content with elongated fatty acids. This quantitative TAG profiling approach should facilitate investigations aimed at understanding the biochemical mechanisms of TAG metabolism in plants.

Patatin-related phospholipase pPLAIIIδ increases seed oil content with long-chain fatty acids in Arabidopsis.

The release of fatty acids from membrane lipids has been implicated in various metabolic and physiological processes, but in many cases, the enzymes involved and their functions in plants remain unclear. Patatin-related phospholipase As (pPLAs) constitute a major family of acyl-hydrolyzing enzymes in plants. Here, we show that pPLAIIIδ promotes the production of triacylglycerols with 20- and 22-carbon fatty acids in Arabidopsis (Arabidopsis thaliana). Of the four pPLAIIIs (α, ß, γ, δ), only pPLAIIIδ gene knockout results in a decrease in seed oil content, and pPLAIIIδ is most highly expressed in developing embryos. The overexpression of pPLAIIIδ increases the content of triacylglycerol and 20- and 22-carbon fatty acids in seeds with a corresponding decrease in 18-carbon fatty acids. Several genes in the glycerolipid biosynthetic pathways are up-regulated in pPLAIIIδ-overexpressing siliques. pPLAIIIδ hydrolyzes phosphatidylcholine and also acyl-coenzyme A to release fatty acids. pPLAIIIδ-overexpressing plants have a lower level, whereas pPLAIIIδ knockout plants have a higher level, of acyl-coenzyme A than the wild type. Whereas seed yield decreases in transgenic plants that ubiquitously overexpress pPLAIIIδ, seed-specific overexpression of pPLAIIIδ increases seed oil content without any detrimental effect on overall seed yield. These results indicate that pPLAIIIδ-mediated phospholipid turnover plays a role in fatty acid remodeling and glycerolipid production.

Understanding pharmacists' experiences with advice-giving in the community pharmacy setting: a focus group study.

OBJECTIVE: To examine the experiences of community pharmacists providing advice about symptoms and complementary and alternative medicines (CAM). METHODS: Ten licensed pharmacists and 21 student pharmacists working in community settings participated in 4 focus groups to discuss: patients' questions about symptoms and CAM, comfort level providing advice, and factors prompting physician referrals. Focus group recordings were transcribed verbatim and interpreted using thematic text analysis. RESULTS: Pharmacists' dual role as advisors and medical liaisons emerged as primary themes. Participants reported that patients often seek their advice about self-care of symptoms to delay physician visits. Participants were comfortable giving advice; lack of medical history decreased their comfort level. Most were uncomfortable recommending CAM because of the lack of regulation and evidence. Participants suggested that pharmacy curricula expand training on symptom triage, pharmacist-patient communication, and CAM to prepare graduates for employment in community settings. CONCLUSION: Student and licensed pharmacists of this study voiced that they are often asked for advice on symptom management, but reported needing training to help provide appropriate advice to patients. PRACTICE IMPLICATIONS: The findings suggest that training strategies could help pharmacists appropriately triage and advise patients seeking self-care advice for their symptoms in the community setting.

Accounting for the sedative and analgesic effects of medication changes during patient participation in clinical research studies: measurement development and application to a sample of institutionalized geriatric patients.

BACKGROUND: To date, no system has been published that allows investigators to adjust for the overall sedative and/or analgesic effects of medications, or changes in medications, in clinical trial participants for whom medication use cannot be controlled. This is common in clinical trials of behavioral and complementary/alternative therapies, and in research involving elderly or chronically ill patients for whom ongoing medical care continues during the trial. This paper describes the development, and illustrates the use, of a method we developed to address this issue, in which we generate single continuous variables to represent the daily sedative and analgesic loads of multiple medications. METHODS: Medications for 90 study participants in a clinical trial of a nonpharmacological intervention were abstracted from medication administration records across multiple treatment periods. An expert panel of three academic clinical pharmacists and a geriatrician met to develop a system by which each study medication could be assigned a sedative and analgesic effect rating. RESULTS: The two measures, when applied to data on 90 institutionalized persons with Alzheimer's disease, resulted in variables with moderately skewed distributions that are consistent with the clinical profile of analgesia and sedation use in long-term care populations. The average study participant received 1.89 analgesic medications per day and had a daily analgesic load of 2.96; the corresponding figures for sedation were 2.07 daily medications and an average daily load of 11.41. CONCLUSIONS: A system of classifying the sedative and analgesic effects of non-study medications was created that divides drugs into categories based on the strength of their effects and assigns a rating to express overall sedative and analgesic effects. These variables may be useful in comparing patients and populations, and to control for drug effects in future studies.

Medication undertreatment in assisted living settings.

BACKGROUND: Residential care/assisted living (RC/AL) is a rapidly growing, long-term care setting, where medication use has not been carefully examined. We sought to determine the prevalence and predictors of nonprescribing of selected medications whose value in decreasing morbidity has been established in clinical trials. METHODS: As part of a survey of a stratified random sample of 193 RC/AL facilities in Florida, Maryland, New Jersey, and North Carolina, data were gathered on 2014 residents 65 years and older. Patient characteristics and diagnoses were recorded based on medical record reviews and in-person patient assessments; all medications administered at least 4 of the previous 7 days were recorded. Data on facility characteristics were obtained by interviewing facility administrators. Bivariate and multivariate logistic regression was performed to identify associations between medication nonprescribing and facility characteristics, physician visitation, and patient age, sex, race, comorbidity, functional dependency, and cognition. RESULTS: Of 328 subjects with congestive heart failure, 204 (62.2%) were not receiving an angiotensin-converting enzyme inhibitor; of 172 subjects with prior myocardial infarction, 60.5% were not receiving aspirin and 76.2% were not receiving beta-blockers; of 435 patients with history of stroke, 37.5% were not receiving an anticoagulant or antiplatelet agent; and of 315 patients with osteoporosis, 61.0% were not receiving calcium supplementation and 51.1% were not receiving any treatment for the condition. Resident age, race, sex, comorbidity, cognitive status, and dependency in activities of daily living were rarely associated with nonprescribing; in contrast, facility factors-particularly facility type and the frequency of physician visits-were somewhat more frequently associated with nonprescribing. CONCLUSIONS: Undertreatment appears to be prevalent in RC/AL facilities. Since preserving independence is often a primary goal of care in these settings, more attention may need to be paid to the use of treatments that have been shown to reduce long-term morbidity.

Evidence for the cardioprotective effects of omega-3 Fatty acids.

OBJECTIVE: To review available literature regarding the cardiovascular effects of marine-derived Omega-3 fatty acids and evaluate the benefit of these fatty acids in the prevention of coronary heart disease. DATA SOURCES: Biomedical literature accessed through a MEDLINE search (1966-April 2002). Search terms included fish oil, omega-3 fatty acid, sudden death, hypertriglyceridemia, myocardial infarction, and mortality. DATA SYNTHESIS: Following an early 1970's observational investigation that Omega-3 fatty acids may reduce the occurrence of myocardial infarction-related deaths in Greenland Eskimos, additional trials have been conducted that support this finding. Epidemiologic and clinical trial data suggest that Omega-3 fatty acids may reduce the risk of cardiovascular-related death by 29-52%. In addition, the risk of sudden cardiac death was found to be reduced by 45-81%. Possible mechanisms for these beneficial effects include antiarrhythmic properties, improved endothelial function, antiinflammatory action, and reductions in serum triglyceride concentrations. Omega-3 Fatty acids are fairly well tolerated; potential adverse effects include bloating and gastrointestinal distress, "fishy taste" in the mouth, hyperglycemia, increased risk of bleeding, and a slight increase in low-density-lipoprotein cholesterol. CONCLUSIONS: Omega-3 Fatty acids may be beneficial and should be considered in patients with documented coronary heart disease. They may be particularly beneficial for patients with risk factors for sudden cardiac death.