The toc132toc120 heterozygote mutant of Arabidopsis thaliana accumulates decreased levels of the major chloroplast lipids.

We used ESI-MS/MS to profile glycerolipids in a mutant of Arabidopsis thaliana that is null and heterozygous for the TOC132 and TOC120 genes, and is referred to as the toc132toc120± mutant. The goal was to assess the impact of a defective atToc132/120 receptor on the accumulation of chloroplast lipids. The mutant accumulated decreased amounts of monogalactosyldiacylglycerol (MGDG), digalactosyldiacylglycerol (DGDG) and phosphatidylglycerol (PG). In the cold-acclimated mutant, PG accumulated at the control levels. However, 34:4-PG (18:3/16:1Δ3trans) was significantly decreased, which indicates that the mutant was impaired in synthesis of the chloroplast-derived PG. Major molecular species of MGDG and DGDG were significantly decreased, which was indicative of the decreased levels of triunsaturated fatty acids in galactolipids. The cold-acclimated mutant accumulated increased levels of phosphatidylcholine (PC), phosphatidylethanolamine (PE) and phosphatidylserine (PS), which indicate that defect in the atToc132/120 receptor did not impair the ER pathway of lipid synthesis. Both cold-acclimated wildtype and mutant plants accumulated increased levels of phosphatidic acid (PA). The increased levels of major molecular species of PA suggest that some pool of PA was derived from degradation of both the chloroplast and extra-chloroplast lipids. The cold-acclimated mutant had decreased double bond index (DBI) and increased acyl chain length (ACL), which was indicative of decreased membrane fluidity. However, a decrease in the ratio of MGDG to DGDG indicate that the mutant was capable of remodeling membrane lipids in response to low temperatures. We conclude that the defective Toc132/120 receptor resulted in decreased synthesis of chloroplast lipids and decreased membrane fluidity.

The TOC159 null mutant of Arabidopsis thaliana is impaired in the accumulation of plastid lipids and phosphatidylcholine.

We used electrospray ionization tandem mass spectrometry to profile glycerolipids in the TOC159 null mutant of Arabidopsis, which is referred to as plastid protein import 2, or ppi2. The goal was to evaluate the impact of a defective atToc159 receptor in the accumulation of plastid lipids. The ppi2 mutant is severely impaired in the accumulation of monogalactosyldiacylglycerol (MGDG), digalactosyldiacylglycerol (DGDG) and phosphatidylglycerol (PG), which are major components of the thylakoid membranes. Major molecular species of MGDG and DGDG are drastically decreased, which is consistent with our previous findings of decreased levels of hexadecatrienoic and linolenic acids. Under normal growth conditions, the ppi2 mutant accumulated significantly lower levels of phosphatidylcholine (PC), phosphatidylethanolamine (PE), and phosphatidylinositol (PI). In the cold-acclimated mutant, the amounts of PE and PI were similar to the wildtype level, which indicates that the ER pathway of lipid synthesis was functional in the mutant. The cold-acclimated ppi2 mutant accumulated increased amounts of phosphatidic acid (PA), which was mirrored by an increase in phospholipase Dα (PLDα) transcript levels. These data suggest that PLDα activity contributed to the accumulation of cold-induced PA in the ppi2 mutant. The accumulation of major molecular species in PA indicates that cold-induced PA originated from the degradation of both plastidial and extraplastidial lipids. Compared with the wildtype, the ppi2 mutant had a low double bond index and high acyl chain length, which is indicative of decreased membrane fluidity. Taken together, these data indicate that a defective atToc159 receptor severely impaired the plastid pathway of lipid synthesis, which negatively affected the synthesis and/or accumulation of PC.

Ozone-induced lipid changes in the wildtype and toc132toc120 heterozygote mutant of Arabidopsis thaliana.

We measured the fatty acids and lipid content in the wildtype and toc132toc120 heterozygote mutant of Arabidopsis thaliana that were exposed to elevated levels of ozone. The goal was to assess whether a defective atToc132/120 receptor would alter the mutant's response to ozone-induced stress. Increased malondialdehyde (MDA) levels were measured in wildtype plants that were exposed to ozone for 3 h and left in an ozone-free environment for 21 h. The increased levels of MDA were not positively correlated with changes in the levels of triunsaturated fatty acids from which MDA is derived. In both the wildtype and mutant plants, absolute amounts of the glycerolipids were not altered by ozone treatment. The untreated mutant, however, accumulated decreased levels of chloroplast lipids and triunsaturated fatty acids. In ozone-treated wildtype, the levels of 16:3 were significantly decreased and this was mirrored by decreased levels of TOC132 and FAD5 transcripts, and increased levels of SP1 E3 ligase transcripts. These data suggest a possible increase in protein ubiquitination under ozone-induced stress. In contrast, in ozone-treated mutant, the FAD5 transcripts accumulated at the control level. The untreated mutant, however, accumulated significantly increased levels of CAT1 and FAD7 transcripts, which indicates that a defective chloroplast receptor induced cellular stress. In ozone-treated wildtype, there was a small increase in 34:6-phosphatidic acid, which indicates that a small amount of the chloroplast-derived MGDG was degraded in response to ozone-induced stress. Overall, these data indicate that the wildtype and mutant responded differently in lipid composition and oxidation to ozone-induced stress.

Safety and feasibility of transcranial direct current stimulation in pediatric hemiparesis: randomized controlled preliminary study.

BACKGROUND: Transcranial direct current stimulation (tDCS) is a form of noninvasive brain stimulation that has shown improved adult stroke outcomes. Applying tDCS in children with congenital hemiparesis has not yet been explored. OBJECTIVE: The primary objective of this study was to explore the safety and feasibility of single-session tDCS through an adverse events profile and symptom assessment within a double-blind, randomized placebo-controlled preliminary study in children with congenital hemiparesis. A secondary objective was to assess the stability of hand and cognitive function. DESIGN: A double-blind, randomized placebo-controlled pretest/posttest/follow-up study was conducted. SETTING: The study was conducted in a university pediatric research laboratory. PARTICIPANTS: Thirteen children, ages 7 to 18 years, with congenital hemiparesis participated. MEASUREMENTS: Adverse events/safety assessment and hand function were measured. INTERVENTION: Participants were randomly assigned to either an intervention group or a control group, with safety and functional assessments at pretest, at posttest on the same day, and at a 1-week follow-up session. An intervention of 10 minutes of 0.7 mA tDCS was applied to bilateral primary motor cortices. The tDCS intervention was considered safe if there was no individual decline of 25% or group decline of 2 standard deviations for motor evoked potentials (MEPs) and behavioral data and no report of adverse events. RESULTS: No major adverse events were found, including no seizures. Two participants did not complete the study due to lack of MEP and discomfort. For the 11 participants who completed the study, group differences in MEPs and behavioral data did not exceed 2 standard deviations in those who received the tDCS (n=5) and those in the control group (n=6). The study was completed without the need for stopping per medical monitor and biostatisticial analysis. LIMITATIONS: A limitation of the study was the small sample size, with data available for 11 participants. CONCLUSIONS: Based on the results of this study, tDCS appears to be safe, feasible, and well tolerated in most children with hemiparesis. Future investigations of serial sessions of tDCS in conjunction with rehabilitation in pediatric hemiparesis are indicated to explore the benefit of a synergistic approach to improving hand function.

Student use and perceptions of different learning aids in a problem-based learning (PBL) dentistry course.

First-year dental students in a new problem-based learning (PBL) course, the Bachelor of Dentistry (BDent) Program at the University of Sydney, Australia, completed the Study Process Questionnaire and two other questionnaires in this study. The study aimed to identify student perceptions of a written formative assessment and the helpfulness of various learning aids used to prepare for this assessment and preparing to be a dental clinician. Correlations between approach to learning and perceptions of assessment and learning aids showed theoretically expected associations. Surface learning was associated with students' concerns regarding whether assessment items reflected curriculum content, a valuing of lectures as a learning aid, and low scores for theme sessions. Deep learning was associated with a perception that the assessment tested application of basic and clinical sciences and a valuing of both independent study groups and learning topics as learning aids. An achievement orientation to learning was associated with a valuing of formative assessment as a learning aid and an intention to modify study habits as a result of participating in formative assessment. The findings provide insight into student learning in a PBL context that will help teachers and curriculum developers better understand the value of teaching aids provided in the program and the impact assessment has on study styles.