Hypothermic retrograde venous perfusion with adenosine cools the spinal cord and reduces the risk of paraplegia after thoracic aortic clamping

AbstractObjective: We evaluated the utility of retrograde venous perfusion to cool the spinal cord and protect neurologic function during aortic clamping. We hypothesized that hypothermic adenosine would preserve the spinal cord during ischemia. Methods: Six swine (group I) underwent thoracic aortic occlusion for 30 minutes at normothermia. Group II animals underwent spinal cooling by retrograde perfusion of the paravertebral veins with hypothermic (4°C) saline solution during aortic occlusion. The spinal cords of group III animals were cooled with a hypothermic adenosine solution in a similar fashion. Intrathecal temperature was monitored and somatosensory evoked potentials assessed the functional status of spinal pathways. Results: Spinal cooling without systemic hypothermia significantly improved neurologic Tarlov scores in group III (4.8 ± 0.2) and group II (3.8 ± 0.4) when compared with group I scores (1.3 ± 0.6) (P < .001). Furthermore, 5 of the 6 animals in group III displayed completely normal neurologic function, whereas only one animal in group II and no animals in group I did (P = .005). Somatosensory evoked potentials were lost 10.6 ± 1.4 minutes after ischemia in group I. In contrast, spinal cooling caused rapid cessation of neural transmission with loss of somatosensory evoked potentials at 6.9 ± 1.2 minutes in group II and 7.0 ± 0.8 minutes in group III (P = .06). Somatosensory evoked potential amplitudes returned to 85% of baseline in group III and 90% of baseline in group II compared with only 10% of baseline in group I (P = .01). Conclusions: We conclude that retrograde cooling of the spinal cord is possible and protects against ischemic injury and that adenosine enhances this effect. The efficacy of this method may be at least partly attributed to a more rapid reduction in metabolic and electrical activity of the spinal cord during ischemia. (J Thorac Cardiovasc Surg 2000;119:588-95

Vegetative buffers for fan emissions from poultry farms: 2. ammonia, dust and foliar nitrogen

This study evaluated the potential of trees planted around commercial poultry farms to trap ammonia (NH3) and dust or particulate matter (PM). Norway spruce, Spike hybrid poplar, hybrid willow, and Streamco purpleosier willow were planted on five commercial farms from 2003 to 2004. Plant foliage was sampled in front of the exhaust fans and at a control distance away from the fans on one turkey, two laying hen, and two broiler chicken farms between June and July 2006. Samples were analyzed for dry matter (DM), nitrogen (N), and PM content. In addition, NH3concentrations were measured downwind of the exhaust fans among the trees and at a control distance using NH3 passive dosi–tubes. Foliage samples were taken and analyzed separately based on plant species. The two layer farms had both spruce and poplar plantings whereas the two broiler farms had hybrid willow and Streamco willow plantings which allowed sampling and species comparisons with the effect of plant location (control vs. fan). The results showed that NH3 concentration h− 1 was reduced by distance from housing fans (P ≤ 0.0001), especially between 0 m (12.01 ppm), 11.4 m (2.59 ppm), 15 m (2.03 ppm), and 30 m (0.31 ppm). Foliar N of plants near the fans was greater than those sampled away from the fans for poplar (3.87 vs. 2.56%; P ≤ 0.0005) and hybrid willow (3.41 vs. 3.02%; P ≤ 0.05). The trends for foliar N in spruce (1.91 vs. 1.77%; P = 0.26) and Streamco willow (3.85 vs. 3.33; P = 0.07) were not significant. Pooling results of the four plant species indicated greater N concentration from foliage sampled near the fans than of that away from the fans (3.27 vs. 2.67%; P ≤ 0.0001). Foliar DM concentration was not affected by plant location, and when pooled the foliar DM of the four plant species near the fans was 51.3% in comparison with 48.5% at a control distance. There was a significant effect of plant location on foliar N and DM on the two layer farms with greater N and DM adjacent to fans than at a control distance (2.95 vs. 2.15% N and 45.4 vs. 38.2% DM, respectively). There were also significant plant species effects on foliar N and DM with poplar retaining greater N (3.22 vs. 1.88%) and DM (43.7 vs. 39.9%) than spruce. The interaction of location by species (P ≤ 0.005) indicated that poplar was more responsive in terms of foliar N, but less responsive for DM than spruce. The effect of location and species on foliar N and DM were not clear among the two willow species on the broiler farms. Plant location had no effect on plant foliar PM weight, but plant species significantly influenced the ability of the plant foliage to trap PM with spruce and hybrid willow showing greater potential than poplar and Streamco willow for PM2.5(0.0054, 0.0054, 0.0005, and 0.0016 mg cm− 2; P ≤ 0.05) and total PM (0.0309, 0.0102, 0.0038, and 0.0046 mg cm− 2, respectively; P ≤ 0.001). Spruce trapped more dust compared to the other three species (hybrid willow, poplar, and Streamco willow) for PM10 (0.0248 vs. 0.0036 mg cm− 2; P ≤ 0.0001) and PM\u3e 10 (0.0033 vs. 0.0003 mg cm− 2; P = 0.052). This study indicates that poplar, hybrid willow, and Streamco willow are appropriate species to absorb poultry house aerial NH3–N, whereas spruce and hybrid willow are effective traps for dust and its associated odors

FY-2007 PNNL Voluntary Protection Program (VPP) Program Evaluation

This document reports the results of the FY-2007 PNNL VPP Program Evaluation, which is a self-assessment of the operational and programmatic performance of the Laboratory related to worker safety and health. The report was compiled by a team of worker representatives and safety professionals who evaluated the Laboratory's worker safety and health programs on the basis of DOE-VPP criteria. The principle elements of DOE's VPP program are: Management Leadership, Employee Involvement, Worksite Analysis, Hazard Prevention and Control, and Safety and Health Training

De novo assembly and transcriptome analysis of five major tissues of Jatropha curcas L. using GS FLX titanium platform of 454 pyrosequencing

<p>Abstract</p> <p>Background</p> <p><it>Jatropha curcas </it>L. is an important non-edible oilseed crop with promising future in biodiesel production. However, factors like oil yield, oil composition, toxic compounds in oil cake, pests and diseases limit its commercial potential. Well established genetic engineering methods using cloned genes could be used to address these limitations. Earlier, 10,983 unigenes from Sanger sequencing of ESTs, and 3,484 unique assembled transcripts from 454 pyrosequencing of uncloned cDNAs were reported. In order to expedite the process of gene discovery, we have undertaken 454 pyrosequencing of normalized cDNAs prepared from roots, mature leaves, flowers, developing seeds, and embryos of <it>J. curcas</it>.</p> <p>Results</p> <p>From 383,918 raw reads, we obtained 381,957 quality-filtered and trimmed reads that are suitable for the assembly of transcript sequences. <it>De novo </it>contig assembly of these reads generated 17,457 assembled transcripts (contigs) and 54,002 singletons. Average length of the assembled transcripts was 916 bp. About 30% of the transcripts were longer than 1000 bases, and the size of the longest transcript was 7,173 bases. BLASTX analysis revealed that 2,589 of these transcripts are full-length. The assembled transcripts were validated by RT-PCR analysis of 28 transcripts. The results showed that the transcripts were correctly assembled and represent actively expressed genes. KEGG pathway mapping showed that 2,320 transcripts are related to major biochemical pathways including the oil biosynthesis pathway. Overall, the current study reports 14,327 new assembled transcripts which included 2589 full-length transcripts and 27 transcripts that are directly involved in oil biosynthesis.</p> <p>Conclusion</p> <p>The large number of transcripts reported in the current study together with existing ESTs and transcript sequences will serve as an invaluable genetic resource for crop improvement in jatropha. Sequence information of those genes that are involved in oil biosynthesis could be used for metabolic engineering of jatropha to increase oil content, and to modify oil composition.</p

Higher Plant Cytochrome b5 Polypeptides Modulate Fatty Acid Desaturation

BACKGROUND: Synthesis of polyunsaturated fatty acids (PUFAs) in the endoplasmic reticulum of plants typically involves the fatty acid desaturases FAD2 and FAD3, which use cytochrome b(5) (Cb5) as an electron donor. Higher plants are reported to have multiple isoforms of Cb5, in contrast to a single Cb5 in mammals and yeast. Despite the wealth of information available on the roles of FAD2 and FAD3 in PUFA synthesis, information regarding the contributions of various Cb5 isoforms in desaturase-mediated reactions is limited. RESULTS: The present functional characterization of Cb5 polypeptides revealed that all Arabidopsis Cb5 isoforms are not similarly efficient in ω-6 desaturation, as evidenced by significant variation in their product outcomes in yeast-based functional assays. On the other hand, characterization of Cb5 polypeptides of soybean (Glycine max) suggested that similar ω-6 desaturation efficiencies were shared by various isoforms. With regard to ω-3 desaturation, certain Cb5 genes of both Arabidopsis and soybean were shown to facilitate the accumulation of more desaturation products than others when co-expressed with their native FAD3. Additionally, similar trends of differential desaturation product accumulation were also observed with most Cb5 genes of both soybean and Arabidopsis even if co-expressed with non-native FAD3. CONCLUSIONS: The present study reports the first description of the differential nature of the Cb5 genes of higher plants in fatty acid desaturation and further suggests that ω-3/ω-6 desaturation product outcome is determined by the nature of both the Cb5 isoform and the fatty acid desaturases

Evolutionary view of acyl-CoA diacylglycerol acyltransferase (DGAT), a key enzyme in neutral lipid biosynthesis

<p>Abstract</p> <p>Background</p> <p>Triacylglycerides (TAGs) are a class of neutral lipids that represent the most important storage form of energy for eukaryotic cells. DGAT (acyl-CoA: diacylglycerol acyltransferase; EC 2.3.1.20) is a transmembrane enzyme that acts in the final and committed step of TAG synthesis, and it has been proposed to be the rate-limiting enzyme in plant storage lipid accumulation. In fact, two different enzymes identified in several eukaryotic species, DGAT1 and DGAT2, are the main enzymes responsible for TAG synthesis. These enzymes do not share high DNA or protein sequence similarities, and it has been suggested that they play non-redundant roles in different tissues and in some species in TAG synthesis. Despite a number of previous studies on the DGAT1 and DGAT2 genes, which have emphasized their importance as potential obesity treatment targets to increase triacylglycerol accumulation, little is known about their evolutionary timeline in eukaryotes. The goal of this study was to examine the evolutionary relationship of the DGAT1 and DGAT2 genes across eukaryotic organisms in order to infer their origin.</p> <p>Results</p> <p>We have conducted a broad survey of fully sequenced genomes, including representatives of Amoebozoa, yeasts, fungi, algae, musses, plants, vertebrate and invertebrate species, for the presence of DGAT1 and DGAT2 gene homologs. We found that the DGAT1 and DGAT2 genes are nearly ubiquitous in eukaryotes and are readily identifiable in all the major eukaryotic groups and genomes examined. Phylogenetic analyses of the DGAT1 and DGAT2 amino acid sequences revealed evolutionary partitioning of the DGAT protein family into two major DGAT1 and DGAT2 clades. Protein secondary structure and hydrophobic-transmembrane analysis also showed differences between these enzymes. The analysis also revealed that the MGAT2 and AWAT genes may have arisen from DGAT2 duplication events.</p> <p>Conclusions</p> <p>In this study, we identified several DGAT1 and DGAT2 homologs in eukaryote taxa. Overall, the data show that DGAT1 and DGAT2 are present in most eukaryotic organisms and belong to two different gene families. The phylogenetic and evolutionary analyses revealed that DGAT1 and DGAT2 evolved separately, with functional convergence, despite their wide molecular and structural divergence.</p

An integrated bioinformatics analysis reveals divergent evolutionary pattern of oil biosynthesis in high- and low-oil plants

Seed oils provide a renewable source of food, biofuel and industrial raw materials that is important for humans. Although many genes and pathways for acyl-lipid metabolism have been identified, little is known about whether there is a specific mechanism for high-oil content in high-oil plants. Based on the distinct differences in seed oil content between four high-oil dicots (20~50%) and three low-oil grasses (<3%), comparative genome, transcriptome and differential expression analyses were used to investigate this mechanism. Among 4,051 dicot-specific soybean genes identified from 252,443 genes in the seven species, 54 genes were shown to directly participate in acyl-lipid metabolism, and 93 genes were found to be associated with acyl-lipid metabolism. Among the 93 dicot-specific genes, 42 and 27 genes, including CBM20-like SBDs and GPT2, participate in carbohydrate degradation and transport, respectively. 40 genes highly up-regulated during seed oil rapid accumulation period are mainly involved in initial fatty acid synthesis, triacylglyceride assembly and oil-body formation, for example, ACCase, PP, DGAT1, PDAT1, OLEs and STEROs, which were also found to be differentially expressed between high- and low-oil soybean accessions. Phylogenetic analysis revealed distinct differences of oleosin in patterns of gene duplication and loss between high-oil dicots and low-oil grasses. In addition, seed-specific GmGRF5, ABI5 and GmTZF4 were predicted to be candidate regulators in seed oil accumulation. This study facilitates future research on lipid biosynthesis and potential genetic improvement of seed oil content