Search

Invasive species are increasingly present in our ecosystems and pose a threat to the health of forest ecosystems. Practitioners are tasked with locating these invasive species and finding ways to mitigate their spread and impacts, often through costly field surveys. Meanwhile, researchers are developing remote sensing products to detect the changes in vegetation health and structure that are caused by invasive species, which could aid in early detection and monitoring efforts. Although both groups are working towards similar goals and field data are essential for validating RS products, these groups often work independently. In this paper, we, a group of researchers and practitioners, discuss the challenges to bridging the gap between researchers and practitioners and summarize the literature on this topic. We also draw from our experiences collaborating with each other to advance detection, monitoring, and management of the Hemlock Woolly Adelgid (Adelges tsugae; HWA), an invasive forest pest in the eastern U.S. We conclude by (1) highlighting the synergies and symbiotic mutualism of researcher–practitioner collaborations and (2) providing a framework for facilitating researcher–practitioner collaborations that advance fundamental science while maximizing the capacity of RS technologies in monitoring and management of complex drivers of forest health decline such as invasive species.

We describe the construction and initial demonstration of a new instructional tool called ROXI (Research Opportunity through eXperimental Instruction). The system interfaces a series of electronic sensors to control software via the Arduino platform. The sensors have been designed to enable low-cost data collection in laboratory courses. Data are collected by a computer and can be displayed or plotted in nearly real time, allowing chemistry to come to life. In addition, student data can be analyzed by the computer automatically and used to provide feedback to assess whether students are analyzing experimental results correctly. Because the computer and software are able to perform all computations and data analysis independently of the student, the software can assess the accuracy of student calculations and even assign grades based upon performance. In addition, since measurement data are logged and plotted, it is possible for the software to assist in assembling laboratory report files that students can...

Several factors determine the dose of drugs in children, but size and age are the most important. Size is most commonly dealt with by weight-based dosing, although surface area is used for some drugs. Young age causes pharmacokinetic and pharmacodynamic differences from adults. Pharmacokinetic changes are due to many factors, but the most important are differences in body composition and immature metabolic pathways. These changes are greatest in neonates, who require smaller doses based on weight than older children. Infants may require larger doses based on weight than all other age groups. Although the metabolism of most drugs is reduced at young ages, suxamethonium, remifentanil and atracurium are exceptions, because their metabolic enzymes are fully active at birth. Sevoflurane is by far the most commonly used volatile agent, and is the only agent available in most countries for inhalational induction. Propofol is an important anesthetic agent in children. It suppresses airway reflexes, reduces emergence delirium and reduces nausea and vomiting. However many drugs, including propofol, are not licensed for all ages and all purposes in children.

BACKGROUND: Thousands of children require mechanical ventilation each year. Although mechanical ventilation is lifesaving, it is also associated with adverse events if not properly managed. The systematic implementation of evidence-based practice through the use of guidelines and protocols has been shown to mitigate risk, yet variation in care remains prevalent. Advances in health-care technology provided the ability to stream data about mechanical ventilation and therapeutic response. Through these advances, a computer system was developed to enable the coupling of physiologic and ventilation data for real-time interpretation. Our aim was to assess the feasibility and utility of a newly developed patient categorization and scoring system to objectively measure compliance with standards of care. METHODS: We retrospectively categorized the ventilation and oxygenation statuses of subjects within our pediatric ICU utilizing 15 rules-based algorithms. Targets were predetermined based on generally accepted practices. All patient categories were calculated and presented as a percent score (0–100%) of acceptable ventilation, acceptable oxygenation, barotrauma-free, and volutrauma-free states. RESULTS: Two hundred twenty-two subjects were identified and analyzed encompassing 1,578 d of mechanical ventilation. Median age was 3 y, median ideal body weight was 14.7 kg, and 63% were male. The median acceptable ventilation score was 84.6%, and the median acceptable oxygenation score was 70.1% (100% being maximally acceptable). Potential for ventilator-induced lung injury was broken into 2 components: barotrauma and volutrauma. There was very little potential for barotrauma, with a median barotrauma-free state of 100%. Median potential for a volutrauma-free state was 56.1%. CONCLUSIONS: We demonstrate the first patient categorization system utilizing a coordinated data-banking system and analytics to determine patient status and a surveillance of mechanical ventilation quality. Further research is needed to determine whether interventions such as visual display of variance from goal and patient categorization summaries can improve outcomes. (ClinicalTrials.gov registration NCT02184208.)

Lake acidification is a major problem in northeastern US lakes that can control fish presence or absence. We examined the history of fish populations in Lake Minnewaska, in eastern New York. We examined historical documents and found that Lake Minnewaska was fishless from 1922-2008 because of high lake-acidity. Following 30 years of recovery from acidic conditions, Notemigonus crysoleucas (Golden Shiner), a small minnow species, was introduced in 2008 and quickly proliferated, peaking at ~15,000 individuals in 2013. In 2012, the piscivorous species Micropterus salmoides (Largemouth Bass) was introduced, and the minnow population was effectively removed by 2014. We present a conceptual model of the history of fish in Lake Minnewaska as fish disappeared and reappeared over 100 years as a consequence of acid rain and human introductions.

BACKGROUND: The systematic implementation of evidence-based practice through the use of guidelines, checklists, and protocols mitigates the risks associated with mechanical ventilation, yet variation in practice remains prevalent. Recent advances in software and hardware have allowed for the development and deployment of an enhanced visualization tool that identifies mechanical ventilation goal variance. Our aim was to assess the utility of daily goal establishment and a computer-aided visualization of variance. METHODS: This study was composed of 3 phases: a retrospective observational phase (baseline) followed by 2 prospective sequential interventions. Phase I intervention comprised daily goal establishment of mechanical ventilation. Phase II intervention was the setting and monitoring of daily goals of mechanical ventilation with a web-based data visualization system (T3). A single score of mechanical ventilation was developed to evaluate the outcome. RESULTS: The baseline phase evaluated 130 subjects, phase I enrolled 31 subjects, and phase II enrolled 36 subjects. There were no differences in demographic characteristics between cohorts. A total of 171 verbalizations of goals of mechanical ventilation were completed in phase I. The use of T3 increased by 87% from phase I. Mechanical ventilation score improved by 8.4% in phase I and 11.3% in phase II from baseline ( P = .032). The largest effect was in the low risk V T category, with a 40.3% improvement from baseline in phase I, which was maintained at 39% improvement from baseline in phase II ( P = .01). mechanical ventilation score was 9% higher on average in those who survived. CONCLUSIONS: Daily goal formation and computer-enhanced visualization of mechanical ventilation variance were associated with an improvement in goal attainment by evidence of an improved mechanical ventilation score. Further research is needed to determine whether improvements in mechanical ventilation score through a targeted, process-oriented intervention will lead to improved patient outcomes. (ClinicalTrials.gov registration NCT02184208.)

The high rates of recurrence and low median survival in many B-cell cancers highlight a need for new targeted therapeutic modalities. In dividing cells, eukaryotic translation initiation factor 5A (eIF5A) is hypusinated and involved in regulation of protein synthesis and proliferation, whereas the non-hypusinated form of eIF5A is a potent inducer of cell death in malignant cells. Here, we demonstrate the potential of modulating eIF5A expression as a novel approach to treating B-cell cancers. SNS01-T is a nonviral polyethylenimine-based nanoparticle, designed to induce apoptosis selectively in B-cell cancers by small interfering RNA–mediated suppression of hypusinated eIF5A and plasmid-based overexpression of a non-hypusinable eIF5A mutant. In this study, we show that SNS01-T is preferentially taken up by malignant B cells, inhibits tumor growth in multiple animal models of B-cell cancers without damaging normal tissues, and synergizes with the current therapies bortezomib and lenalidomide to inhibit tumor progression. The results collectively demonstrate the potential of SNS01-T as a novel therapeutic for treatment of a diverse range of B-cell malignancies.

AteIF5A3, one of three genes encoding eukaryotic translation initiation factor 5A (eIF5A) in Arabidopsis thaliana, and corresponding genes PdeIF5A3 from Populus deltoides (eastern cottonwood) and SleIF5A4 from Solanum lycopersicum (tomato) were constitutively over-expressed in A. thaliana. The resultant transgenic plants exhibited enhanced vegetative and reproductive growth. Indeed, the increase in seed yield relative to empty vector controls for the PdeIF5A3 over-expressing plants ranged from 50% to 300% depending on the line. The PdeIF5A3 over-expressing plants also exhibited enhanced fitness when exposed to osmotic and nutrient (N, P and K) stress. The spatial localization of AteIF5A3 was visualized by confocal microscopy using transgenic plants expressing P(AteIF5A3) :GFP-AteIF5A3. GFP fluorescence reflecting expression of AteIF5A3 was detectable in the phloem, particularly companion cells, of roots, stems and leaves, in the epidermal cells of the root tip, in the columella cells of the root cap and in the chalazal tissue of fertilized ovules, which all play a pivotal role in nutrient or hormone translocation. Thus, AteIF5A3 appears to be involved in supporting growth and to play a regulatory role in the response of plants to sub-lethal osmotic and nutrient stress.

Reliable, long-term records of daily weather and climate are relatively rare but are crucial for understanding long-term trends and variability in extreme events and other climate metrics that are not resolvable at the monthly time scale. Here, the distinct features of a continuous, long-term (1896–2006) daily weather record from Mohonk Lake, New York, are highlighted. The site is optimal for daily climate analyses, since it has experienced negligible land-use change, no station moves, and has maintained methodological and instrumental consistency over the entire period of record. Unlike many sites, the site has always used maximum/minimum thermometers rather than shifting to the automated Maximum/Minimum Temperature Sensor. Notable results from the analysis of this record include 1) a warming trend driven largely by trends in maximum temperatures, especially during summer, 2) increasing diurnal temperature range during summer, and 3) a reduction in the number of freeze-days per year with little change in the length of the freeze-free season. These findings deviate from some regional level trends, suggesting there may be value in revisiting selected, consistently monitored, and maintained stations similar to Mohonk for focused analyses of regional climate change.

To identify the effect of perflubron on gas exchange and lung mechanics during high frequency oscillatory ventilation in an animal model.Prospective randomized animal trial.Eighteen Yorkshire swine.Three groups of six animals each were investigated: control (high frequency oscillatory ventilation alone), low dose perflubron (high frequency oscillatory ventilation plus perfluoro-octyl bromide [PFOB]-Lo, 1.5 mL/kg), and high dose perflubron (high frequency oscillatory ventilation plus PFOB-Hi, 3 mL/kg). Lung injury was induced with repeated saline lavage and amplified for 1 hr using large tidal volumes. Perflubron (Alliance, CA) or a sham dose (room air) was administered with bronchoscopic guidance. The animals were transitioned to high frequency oscillatory ventilation starting at a mean airway pressure of 15 cm H2O. Mean airway pressure was increased (inflation phase) by 5 cm H2O every 15 mins to a maximum mean airway pressure of 40 cm H2O. During the deflation phase, mean airway pressure was reduced by 5 cm H2O every 15 mins to a mean airway pressure of 15 cm H2O.Oxygenation was improved and pulmonary shunt fraction was reduced for PFOB-Hi compared with the control group only at a mean airway pressure of 15 and 20 cm H2O. At a maximal mean airway pressure of 40 cm H2O, oxygenation was not different between the groups, but pulmonary artery pressures were elevated in both PFOB-groups compared with the control group. During the deflation phase, oxygenation, pulmonary shunt fraction, and pulmonary artery pressures were adversely affected by PFOB-Hi and PFOB-Lo.Although PFOB-Hi compared with the control group improved oxygenation and reduced pulmonary shunt fraction only during the first pressure steps of a formal stepwise recruitment maneuver during high frequency oscillatory ventilation, this effect was not sustained during maximal recruitment. During the deflation phase, both PFOB groups were associated with worse gas exchange compared with the control group. PFOB also produced significant pulmonary hypertension in comparison with the control group.

We present a detailed analysis of a rare cross-taxa native species phenology dataset (plant flowering, insect first sighting, and amphibian first sighting) from Mohonk Lake, NY. This dataset is highly unusual in North America for its longevity of record, consistency of methodology and location, diversity of species available, and availability of local daily meteorological data. For each phenology series, we examined the flowering and first sighting Julian calendar dates for the existence of temporal trends. Only one of the five animal species (katydid) showed any evidence for a significant trend in first sighting. In contrast, the plant species showed a rich mixture of temporal trends in flowering that could be divided into four classes: woody plant–no trend, woody plant–negative trend, herbaceous plant–negative trend, and herbaceous plant–positive trend. Many of the trends were found to be statistically significant and robust to the method of trend estimation. The data within each of the four plant classes were also pooled as anomalies to provide more complete temporal coverage for tests of trend robustness. The results were strongly consistent with the trends of the individual species within each class and highly significant statistically. We next correlated the flowering and first sighting dates against growing degree-day (GDD) summations for each day of the year to measure the sensitivity of each species to this common form of climatic forcing on phenology. All species showed a significant sensitivity to GDD summations, with peak correlations falling on or near the varying median flowering or first sighting dates. These results were robust whether the GDD analyses were conducted over the complete set of observations for each species (beginning on or after 1928) or over a latter period with a more serially complete set of cross-taxa observations (1970–2002). The GDD correlations indicate significant climate sensitivity in all species, but the different magnitudes of correlation and timing of maximum correlation imply that plant and animal responses to climate changes in the future will not be homogeneous for the tested species at Mohonk Lake. Copyright © 2007 Royal Meteorological Society

Eukaryotic translation initiation factor 5A (eIF5A) is a highly conserved protein found in all eukaryotic kingdoms. This study demonstrates that plant eIF5A is involved in the development of disease symptoms induced by a common necrotrophic bacterial phytopathogen. Specifically, AteIF5A-2, one of the three eIF5A genes in Arabidopsis (Arabidopsis thaliana), is shown to regulate programmed cell death caused by infection with virulent Pseudomonas syringae pv tomato DC3000 (Pst DC3000). Transgenic Arabidopsis plants with constitutively suppressed AteIF5A-2 exhibited marked resistance to programmed cell death induced by virulent Pst DC3000, and there was a corresponding reduction in pathogen growth and development of disease symptoms in the plant tissue. Constitutive overexpression of AteIF5A-2 circumvented the apparent posttranscriptional regulation of AteIF5A-2 protein expression characteristic of wild-type plants but did not increase susceptibility to virulent Pst DC3000 ingression. The transgenic plants with constitutive AteIF5A-2 overexpression did, however, display phenotypes consistent with precocious cell death. The results indicate that AteIF5A-2 is a key element of the signal transduction pathway resulting in plant programmed cell death.

Senescence is a highly orchestrated developmental stage in the life cycle of plants. The onset of senescence is tightly controlled by signaling cascades that initiate changes in gene expression and the synthesis of new proteins. This complement of new proteins includes hydrolytic enzymes capable of executing catabolism of macromolecules, which in turn sets in motion disassembly of membrane molecular matrices, leading to loss of cell function and, ultimately, complete breakdown of cellular ultrastructure. A distinguishing feature of senescence that sets it apart from other types of programmed cell death is the recovery of carbon and nitrogen from the dying tissue and their translocation to growing parts of the plant such as developing seeds. For this to be accomplished, the initiation of senescence and its execution have to be meticulously regulated. For example, the initiation of membrane disassembly has to be intricately linked with the recruitment of nutrients because their ensuing translocation out of the senescing tissue requires functional membranes. Molecular mechanisms underlying this linkage and its integration with the catabolism of macromolecules in senescing tissues are addressed.

Purpose This paper describes the methodology of a clinical trial of prone positioning in pediatric patients with acute lung injury (ALI). Nonrandomized studies suggest that prone positioning improves oxygenation in patients with ALI/acute respiratory distress syndrome without the risk of serious iatrogenic injury. It is not known if these improvements in oxygenation result in improvements in clinical outcomes. A clinical trial was needed to answer this question. Materials and Methods The pediatric prone study is a multicenter, randomized, noncrossover, controlled clinical trial. The trial is designed to test the hypothesis that at the end of 28 days, children with ALI treated with prone positioning will have more ventilator-free days than children treated with supine positioning. Secondary end points include the time to recovery of lung injury, organ failure–free days, functional outcome, adverse events, and mortality from all causes. Pediatric patients, 42 weeks postconceptual age to 18 years of age, are enrolled within 48 hours of meeting ALI criteria. Patients randomized to the prone group are positioned prone within 4 hours of randomization and remain prone for 20 hours each day during the acute phase of their illness for a maximum of 7 days. Both groups are managed according to ventilator protocol, extubation readiness testing, and sedation protocols and hemodynamic, nutrition, and skin care guidelines. Conclusions This paper describes the process, multidisciplinary input, and procedures used to support the design of the clinical trial, as well as the challenges faced by the clinical scientists during the conduct of the clinical trial.

Deoxyhypusine synthase (DHS; EC 2.5.1.46) mediates the first of two enzymatic reactions that convert inactive eukaryotic translation initiation factor-5A (eIF-5A) to an activated form, thought to facilitate translation. A full-length cDNA clone encoding canola (Brassica napus cv. Westar) DHS was isolated from a cDNA-expression library prepared from senescing leaves. Transgenic canola lines with suppressed DHS expression were obtained by introducing a transgene expressing antisense 3′-UTR canola DHS cDNA under the regulation of the constitutive cauliflower mosaic virus 35S (CaMV-35S) promoter. Transformed seed was obtained by vacuum infiltration of canola inflorescences using the protocol developed for Arabidopsis with modifications. The resultant transgenic plants had reduced levels of leaf DHS protein and exhibited delayed natural leaf senescence. Suppression of DHS also increased leaf size by 1.5- to two-fold and resulted in increases in seed yield of up to 65%. Moreover, the enhanced performance of transgenic plants reflected increased tolerance to chronic sublethal stress. When wild-type and transgenic plants were grown in 6-inch pots, the increase in seed yield accruing from suppression of DHS was approximately 4.5-fold greater than when the plants were grown in 12-inch pots. Thus, suppression of DHS appears to ameliorate the effects of sublethal stress engendered by growth in small containers.

The effects of suppressing deoxyhypusine synthase (DHS) have been examined in tomato (Solanum lycopersicum cv UCT5). DHS mediates the first of two sequential enzymatic reactions that activate eukaryotic translation initiation factor-5A (eIF-5A) by converting a conserved Lys to the unusual amino acid, deoxyhypusine. DHS protein levels were suppressed in transgenic plants by expressing the 3′-untranslated region of tomato DHS under regulation of the constitutive cauliflower mosaic virus promoter. Fruit from the transgenic plants ripened normally, but exhibited delayed postharvest softening and senescence that correlated with suppression of DHS protein levels. Northern-blot analysis indicated that all four gene family members of tomato eIF-5A are expressed in fruit, and that three are up-regulated in parallel with enhancement of DHS mRNA as the fruit begin to senesce and soften. Transgenic plants in which DHS was more strongly suppressed were male sterile, did not produce fruit, and had larger, thicker leaves with enhanced levels of chlorophyll. The activity of PSII was 2 to 3 times higher in these transgenic leaves than in corresponding leaves of wild-type plants, and there was also enhanced deposition of starch in the stems. The data collectively indicate that suppression of DHS has pleiotropic effects on growth and development of tomato. This may, in turn, reflect the fact that there is a single DHS gene in tomato and that its cognate protein is involved in the activation of four distinct isoforms of eIF-5A.

Evidence suggests that glycemic control provides clinical benefit to critically ill patients. The Extracorporeal Glucose Monitoring System (EGMS), Medtronic Minimed, Northridge, CA) has been developed to measure real-time, continuous blood glucose concentrations in patients on extracorporeal bypass. This pilot study reports the first in vitro and in vivo evaluations of EGMS in an extracorporeal circuit.In an in vitro study, three EGMS sensors were inserted in a neonatal extracorporeal circuit. Circuit blood glucose levels were altered by saline dilution and dextrose infusion. EGMS sensors were then inserted into the venous return limb of the extracorporeal circuit in a cohort of six critically ill infants on extracorporeal life support (ECLS). Continuous glucose measurements were compared with laboratory and bedside glucose values at predefined intervals. Linear regression analyses and the Clarke error grid were constructed to analyze device accuracy.All three in vitro EGMS sensors recorded real-time data without interruption for 48 h. EGMS glucose measurements closely correlated with reference levels (R (2) = 0.93). EGMS glucose values demonstrated an approximate 7-10 min lag while glucose concentrations were rapidly changing. Eight EGMS devices were inserted into six neonates on ECLS on day of life 6 +/- 3. EGMS correlated well with laboratory glucose (R2 = 0.61) and bedside glucose during a hyperinsulinemic euglycemic clamp (R2 = 0.78). On the Clarke error grid, 98% of readings were within zones A and B using laboratory glucose as reference, and 100% were within zones A and B using bedside glucose measurements. Blood glucose range during the in vitro study was 19-295 mg/dL and during the in vivo study was 80-257 mg/dL.This pilot study suggests that EGMS is a reliable tool for measuring continuous blood glucose in critically ill patients connected to an extracorporeal circuit, although important limitations exist. Potential applications of this technology include intensive glucose monitoring in patients on ECLS, cardiopulmonary bypass, and renal replacement therapy.

Regulation of protein synthesis is increasingly being recognized as an important determinant of cell proliferation and senescence. In particular, recent evidence indicates that eukaryotic translation initiation factor 5A (eIF-A) plays a pivotal role in this determination. Separate isoforms of eIF-5A appear to facilitate the translation of mRNAs required for cell division and cell death. This raises the possibility that eIF-5A isoforms are elements of a biological switch that is in one position in dividing cells and in another position in dying cells. Changes in the position of this putative switch in response to physiological and environmental cues are likely to have a significant impact on plant growth and development.

High-frequency oscillatory ventilation with perflubron (PFB) reportedly improves pulmonary mechanics and gas exchange and attenuates lung injury. We explored PFB evaporative loss kinetics, intrapulmonary PFB distribution, and dosing strategies during 15 h of high-frequency oscillation (HFO)-partial liquid ventilation (PLV). After saline lavage lung injury, 15 swine were rescued with high-frequency oscillatory ventilation ( n = 5), or in addition received 10 ml/kg PFB delivered to dependent lung [ n = 5, PLV-compartmented (PLV(C))] or 10 ml/kg distributed uniformly within the lung [ n = 5, PLV(U)]. In the PLV(C) group, PFB vapor loss was replaced. ANOVA revealed an unsustained improvement in oxygenation index in the PLV(U) group ( P = 0.04); the reduction in oxygenation index correlated with PFB losses. Although tissue myeloperoxidase activity was reduced globally by HFO-PLV ( P < 0.01) and regional lung injury scores (lung injury scores) in dependent lung were improved ( P = 0.05), global lung injury scores were improved by HFO-PLV ( P < 0.05) only in atelectasis, edema, and alveolar distension but not in cumulative score. In our model, markers of inflammation and lung injury were attenuated by HFO-PLV, and it appears that uniform intrapulmonary PFB distribution optimized gas exchange during HFO-PLV; additionally, monitoring PFB evaporative loss appears necessary to stabilize intrapulmonary PFB volume.

A full-length cDNA clone encoding deoxyhypusine synthase (DHS) was isolated from a cDNA expression library prepared from senescing leaves of Arabidopsis thaliana. Southern blot analysis indicated that DHS is encoded by a single-copy gene in Arabidopsis. During leaf development, the abundance of DHS mRNA in the third pair of rosette leaves peaked at days 14 and 35 after emergence coincident with the initiation of bolting and the later stages of leaf senescence, respectively. These changes in DHS expression were paralleled by corresponding changes in transcript abundance for eIF-5A1, one of three isoforms of eIF-5A in Arabidopsis. Levels of DHS transcript also increased in detached leaves coincident with post-harvest senescence. DHS was suppressed in transgenic plants by introducing antisense full-length or 3'-untranslated Arabidopsis DHS cDNA under the regulation of the constitutive cauliflower mosaic virus (CaMV-35S) promoter. Plants expressing the antisense transgenes had reduced levels of leaf DHS protein and, depending on the level of DHS suppression, exhibited delayed natural leaf senescence, delayed bolting, increased rosette leaf and root biomass, and enhanced seed yield. Suppression of DHS also delayed premature leaf senescence induced by drought stress resulting in enhanced survival in comparison with wild-type plants. In addition, detached leaves from DHS-suppressed plants exhibited delayed post-harvest senescence. These pleiotropic effects of DHS suppression indicate that the protein plays a central role in plant development and senescence.

Lipid analysis of rosette leaves from Arabidopsis has revealed an accumulation of triacylglycerol (TAG) with advancing leaf senescence coincident with an increase in the abundance and size of plastoglobuli. The terminal step in the biosynthesis of TAG in Arabidopsis is catalyzed by diacylglycerol acyltransferase 1 (DGAT1; EC 2.3.1.20). When gel blots of RNA isolated from rosette leaves at various stages of development were probed with the Arabidopsis expressed sequence tag clone, E6B2T7, which has been annotated as DGAT1, a steep increase in DGAT1 transcript levels was evident in the senescing leaves coincident with the accumulation of TAG. The increase in DGAT1 transcript correlated temporally with enhanced levels of DGAT1 protein detected immunologically. Two lines of evidence indicated that the TAG of senescing leaves is synthesized in chloroplasts and sequesters fatty acids released from the catabolism of thylakoid galactolipids. First, TAG isolated from senescing leaves proved to be enriched in hexadecatrienoic acid (16:3) and linolenic acid (18:3), which are normally present in thylakoid galactolipids. Second, DGAT1 protein in senescing leaves was found to be associated with chloroplast membranes. These findings collectively indicate that diacylglycerol acyltransferase plays a role in senescence by sequestering fatty acids de-esterified from galactolipids into TAG. This would appear to be an intermediate step in the conversion of thylakoid fatty acids to phloem-mobile sucrose during leaf senescence.

Neurolinguistic programming (NLP) is a set of techniques designed to make you better at what you do. NLP helps you to understand yourself and others and teaches you how to communicate effectively with others in order to build better relationships with them. Reports on a longitudinal study of the effect of a two to three day NLP course on managers and staff in the hospitality industry. The evaluation is unique as it is the only known longitudinal research into the effects of NLP. Measurements were taken on four psychometric measures at four time periods. The four constructs measured are all positively linked to performance. Three show increases over the start of course measure. Self‐efficacy does not. The verdict on the efficacy of NLP is unclear. Draws some conclusions on the extension of the research.

Full-length cDNA clones encoding deoxyhypusine synthase (DHS) and eucaryotic initiation factor 5A (eIF-5A) have been isolated from a cDNA expression library prepared from tomato leaves (Lycopersicon esculentum, cv. Match) exposed to environmental stress. DHS mediates the first of two enzymatic reactions that activate eIF-5A by converting a conserved lysine to the unusual amino acid, deoxyhypusine. Recombinant protein obtained by expressing tomato DHS cDNA in Escherichia coli proved capable of carrying out the deoxyhypusine synthase reaction in vitro in the presence of eIF-5A. Of particular interest is the finding that DHS mRNA and eIF-5A mRNA show a parallel increase in abundance in senescing tomato flowers, senescing tomato fruit, and environmentally stressed tomato leaves exhibiting programmed cell death. Western blot analyses indicated that DHS protein also increases at the onset of senescence. It is apparent from previous studies with yeast and mammalian cells that hypusine-modified eIF-5A facilitates the translation of a subset of mRNAs mediating cell division. The present study provides evidence for senescence-induced DHS and eIF-5A in tomato tissues that may facilitate the translation of mRNA species required for programmed cell death.

Objective: Partial liquid ventilation with the perfluorochemical, perflubron, has been shown to improve lung mechanics and enhance gas exchange in the treatment of severe acute lung injury. However, the most effective strategy to provide optimal intrapulmonary distribution of perflubron has not been fully accessed. The objective of this study was to examine the effect of body position (supine vs. rotational) and mode of ventilation (conventional mechanical ventilation [CMV] vs. high-frequency oscillatory ventilation [HFOV]) on perflubron distribution and oxygenation improvement. Design: Prospective, randomized, animal trial. Setting: Research laboratory at a university medical center. Subjects: Twenty healthy piglets (4.5-6.6 kg). Interventions: Subjects underwent repetitive saline lavage to achieve a uniform degree of lung injury and then were randomized to either CMV or were converted to HFOV. Within each ventilator group, animals were randomized to supine positioning (S) or rotational positioning with alternation between supine and prone position (R) during incremental dosing of three 5-mUkg doses of perflubron. Measurements and Main Results: Arterial blood gas tensions, hemodynamic variables, and the oxygenation index were recorded after each dose of 5 mUkg. Lateral cinefluoroscopic images after each dose were digitized for computer analysis of density. A density index was calculated for a 2-cm 2 window in three dorsal and three ventral lung regions. Uniformity of distribution was calculated by comparing the mean density among the six regions. Oxygenation improvements were compared between groups. There were no significant differences in hemodynamic variables or gas exchange after lung injury in the four groups. Rotational positioning produced significantly more uniform perflubron distribution during both CMV and HFOV. This effect was independent of the mode of ventilation. The mean ventral density index was affected by rotating position and HFOV mode of ventilation after 10 mUkg of perflubron, and rotating position was affected only after 15 mUkg of perflubron. There was a significant reduction in the oxygenation index from baseline to end lavage in both CMV groups, as well as all of the animals that were rotated. Conclusion: Perflubron is more uniformly dispersed when dosed in a rotational fashion with alternation between supine and prone position during incremental dosing. This effect is independent of mode of ventilation. There was no relationship between oxygenation improvements and nondependent perflubron distribution. CMV and rotating dosing both led to a significant decrease in the oxygenation index after a 15 mUkg dose of perflubron. This information has important impact on the future development of dosing strategies and clinical trial design.

Increasing demands in university management in the 1990s in the UK requires a re‐examination of the head of school’s role. The Standards are part of a radical change in the system of vocational education and training in the UK which began in 1982. A questionnaire was distributed to deans and senior officers, heads and staff. Respondents gave their ratings of importance of 62 activities in the effective leading and managing of a school derived from the Standards. In feedback workshops heads and deans commented on changes to the proposed model. The validity of the standards and their future usefulness is evidenced by few changes to the proposed model and willingness by heads and deans to use the model as the platform for future development.

We previously reported improved oxygenation, but no change, in rates of extracorporeal membrane oxygenation (ECMO) use or death among infants with persistent pulmonary hypertension of the newborn who received inhaled nitric oxide (NO) with conventional ventilation, irrespective of lung disease. The goal of our study was to determine whether treatment with inhaled NO improves oxygenation and clinical outcomes in infants with persistent pulmonary hypertension of the newborn and associated lung disease who are ventilated with high-frequency oscillatory ventilation (HFOV).Single-center, prospective, randomized, controlled trial.Newborn intensive care unit of a tertiary care teaching hospital.We studied infants with a gestational age ofor =34 wks who were receiving mechanical ventilatory support and had echocardiographic and clinical evidence of pulmonary hypertension and hypoxemia (PaO2or =100 mm Hg on FIO2 = 1.0), despite optimal medical management Infants with congenital heart disease, diaphragmatic hernia, or other major anomalies were excluded.The treatment group received inhaled NO, whereas the control group did not. Adjunct therapies and ECMO criteria were the same in the two groups of patients. Investigators and clinicians were not masked as to treatment assignment, and no crossover of patients was permitted.Primary outcome variables were mortality and use of ECMO. Secondary outcomes included change in oxygenation and duration of mechanical ventilatory support and supplemental oxygen therapy. Forty-two patients were enrolled. Baseline oxygenation and clinical characteristics were similar in the two groups of patients. Infants in the inhaled NO group (n = 21) had improved measures of oxygenation at 15 mins and 1 hr after enrollment compared with infants in the control group (n = 20). Fewer infants in the inhaled NO group compared with the control group were treated with ECMO (14% vs. 55%, respectively; p = .007). Mortality did not differ with treatment assignment.Among infants ventilated by HFOV, those receiving inhaled NO had a reduced need for ECMO. We speculate that HFOV enhances the effectiveness of inhaled NO treatment in infants with persistent pulmonary hypertension of the newborn and associated lung disease.

The sonolytic inactivation of Escherichia coli ( E. coli ) is reported. The impact of power intensity, dissolved gas and ultrasonic frequency on the germicide effectiveness of sonification has been explored. The inactivation of E. coli exhibits pseudo-first order behavior, and depends moderately on total power and power intensity at a frequency of 20 kHz. The rate coefficients in an oxygenated solution vary from 0.031 to 0.046 min −1 when the power intensity ranges from 4.6 to 74 W cm −2 . Inactivation occurs most readily at the highest sound intensity, 74 W cm −2 . Three dissolved gases were investigated: oxygen, argon, and an argon/oxygen mixture. The nature of the dissolved gas does not strongly influence the magnitude of the inactivation coefficients which varied from k =0.027 min −1 to 0.047 min −1 . Ultrasonic frequency, within the limits of 205–1017 kHz, displays a strong influence on the rate of E. coli inactivation in oxygenated solutions. The rate coefficients vary from 0.030 to 0.078 min −1 . The most effective ultrasonic frequency for E. coli innovation is 205 kHz; the rate coefficient ( k =0.078 min −1 ) is greater than the coefficient at 1071 kHz ( k =0.030 min −1 ) by a factor of 2.6 and 358 kHz ( k =0.064 min −1 ) also demonstrates a greater efficiency than either 618 kHz ( k =0.041 min −1 ) or 1017 kHz.

Distinguishable populations of lipid particles isolated from chloroplasts of yellow wax bean (Phaseolus vulgaris L. cv Kinghorn Wax) leaves have been found to contain plastid-lipid-associated protein (J. Pozueta-Romero, F. Rafia, G. Houlné, C. Cheniclet, J.P. Carde, M.-L. Schantz, R. Schantz [1997] Plant Physiol 115: 1185–1194). One population is comprised of plastoglobuli obtained from sonicated chloroplasts by flotation centrifugation. Higher density lipid-protein particles isolated from chloroplast stroma by ultrafiltration constitute a second population. Inasmuch as the stromal lipid-protein particles contain plastid-lipid-associated protein, but are distinguishable from plastoglobuli in terms of their lipid and protein composition, they appear to be plastoglobuli-like particles. Of particular interest is the finding that plastoglobuli and the higher density lipid-protein particles both contain catabolites of the thylakoid protein, cytochromef. These observations support the view that there are distinguishable populations of plastoglobuli-like particles in chloroplasts. They further suggest that the formation of these particles may allow removal of protein catabolites from the thylakoid membrane that are destined for degradation as part of normal thylakoid turnover.

Wastewater and drinking water disinfection are typically achieved via chlorination, ozonation, or UV irradiation. However, there has been increased interest in recent years in alternative disinfectants. This interest came about primarily as a result of concerns over the toxicological effects of disinfection by-products created by conventional disinfection processes and the resistance of some recalcitrant microorganisms to these disinfectants. The work reported herein represents an investigation of the effect of an alternative disinfectant, gamma radiation, on the viability of three important waterborne microorganisms. \IEscherichia coli\N, coliphage MS-2, and \ICryptosporidium parvum\N were chosen for this investigation as representative bacterial, viral, and protozoan microorganisms, respectively. A \u6\u0Co irradiator was used to expose test microorganisms to a controlled radiation dose. Experiments were performed for each of the test microorganisms to evaluate the effect of dissolved oxygen concentration and carbonate alkalinity on inactivation efficiency. For each microorganism, a strong effect of dissolved oxygen was observed, regardless of alkalinity. A subtle effect of alkalinity was observed for \IE. coli\N and coliphage MS-2, but only in air-saturated solutions. No significant alkalinity effect was observed for \ICryptosporidium parvum\N. Inactivation kinetics were modeled for \IE. coli\N and coliphage MS-2 using single-target theory to calculate an inactivation rate constant. Multitarget theory was used to represent the inactivation response of \ICryptosporidium parvum\N. The inactivation models based on target theory were found to provide suitable representations of experimental observations.

Study objective To describe the physiologic changes and to evaluate the safety of placing pediatric patients with acute lung injury (ALI) prone for 20 h/d during the acute phase of their illness. Design Single-center prospective case series. Setting Tertiary-level pediatric ICU. Patients Consecutive patients with bilateral pulmonary parenchymal disease requiring intubation and mechanical ventilation with a Pao 2 /fraction of inspired oxygen (Fio 2 ) ratio ≤ 300 mm Hg. Interventions Patients were enrolled as soon as possible after meeting criteria and were placed in a prone position for 20 h/d daily until clinical improvement or death occurred. Measurements and results Twenty-five pediatric patients who had ali/ards, ranging in age from 2 months to 17 years, were placed in a prone position within 19 h of meeting the study criteria for a median time of 4 days, which accounted for 47% of their time receiving mechanical ventilation. Eighty-four percent of patients (n = 21) were categorized as overall responders to prone positioning because they experienced more days of increases of ≥ 20 mm Hg in Pao 2 /Fio 2 ratio or a decrease of ≥ 10% in oxygenation index when shifted from a supine to a prone position during the study period. During the 107 patient-days and 214 positioning cycles, no critical incidents occurred. Furthermore, no patient experienced a persistent decrease in oxygen saturation as measured by pulse oximetry (Spo 2 ) of > 10% from values obtained when in the supine position, failed to keep their Spo 2 at > 85%, or experienced an increased respiratory rate of > 40 breaths/min when prone. Using the comfort score, patients were objectively rated to be equally comfortable in both the supine and prone positions. Patients also were able to resume spontaneous ventilation and to progress toward endotracheal extubation while in the prone position. Iatrogenic injury associated with prolonged prone positioning included stage ii pressure ulcers in six patients (24%). Conclusions The pediatric patients in this series demonstrated improvements in oxygenation without serious iatrogenic injury after prone positioning. This study provides a foundation for a prospective randomized study investigating the effect of early and repeated prone positioning on clinical outcomes in pediatric patients with ALI.

Lipid-protein particles originating from the plasma membrane were immunopurified from the cytosol of carnation petal cells (Dianthus caryophyllus L. cv. Improved White Sim) using antibodies raised against the central hydrophilic domain of the H + -ATPase. The immunopurified particles are enriched in lipid metabolites, in particular free fatty acids and steryl/wax esters, by comparison with corresponding microsomal membranes, and the lipids of the particles are more saturated than those of microsomal membranes. Proteolytic catabolites of the H + -ATPase, a protein associated with the plasma membrane, but not the native H + -ATPase protein, are also present in the immunopurified cytosolic particles. Osmiophilic particles were discernible in the cytosol of carnation petal cells by transmission electron microscopy, and the association of H + -ATPase catabolites with a subpopulation of these particles was confirmed by immunogold labelling with H + -ATPase antiserum. Cross-reaction of the H + -ATPase antiserum with elements of the cytosol was also evident by immunofluorescent light microscopy. These observations collectively indicate that lipid-protein particles of plasma membrane origin are present in the cytosol of carnation petal cells and that their formation may serve as a means of removing lipid and protein metabolites from the plasma membrane which would otherwise destabilize its structure.

Objective: To examine the utility of single-breath CO 2 analysis as a measure of lung volume change in a model of acute lung injury. Setting: Animal laboratory in a university-affiliated medical center. Design: Prospective, animal cohort study comparing 21 variables derived from single-breath CO 2 analysis with lung volume measurements determined by nitrogen washout. Subjects: Seven lambs with saline lavage-induced acute lung injury. Methods: Animals were treated with repetitive saline lavage to achieve a uniform degree of acute lung injury (Pao 2 < 100 torr [13.32 kPa] on Flo 2 of 1.0). Twenty-one derived components of the CO 2 expirogram were evaluated as predictors of lung volume change. Lung volume was manipulated by 3-cm H 2 O incremental increases in positive end-expiratory pressure from 0 to 21 cm H 2 O and ranged between 90 and 765 mL. Measurements and Main Results: Fifty-five measurements of lung volume were available for comparison with derived variables from the CO 2 expirogram. Stepwise linear regression identified five variables that were most predictive of lung volume change: a) dynamic lung compliance; b) the slope of phase III; c) the slope of phase II divided by the mixed expired CO 2 concentration; d) airway deadspace; and e) Pao 2 /Flo 2 ratio. The multivariate equation was highly statistically significant and explained 94% of the variance (adjusted r 2 =.94, p

In recent years there has been an increase in the interest in the production of compounds by isolation from natural sources or through processes that can be deemed "natural". This is of particular interest in the food and beverage industry for flavors and aromas. Hexanal, organoleptically known to possess "green character", is of considerable commercial interest. The objective of this study was to determine if the enzyme template known to be responsible for the synthesis of hexanal from linoleic acid (18:2) in tomato fruits could be harnessed using a hollow-fiber reactor. A hollow-fiber reactor system was set up and consisted of a XAMPLER ultrafiltration module coupled to a reservoir. The enzyme template was extracted from ripe tomato fruits and processed through an ultrafiltration unit (NMWC of 100 kDa) to produce a retentate enriched in soluble and membrane-associated lipoxygenase (LOX) and hydroperoxide lyase (HPL). This extract was recirculated through the lumen of the hollow-fiber ultrafiltration unit with the addition of substrate in the form of linoleic acid, with buffer addition to the reaction flask to maintain a constant retentate volume. Product formation was measured in the permeate using solid phase microextraction (SPME) developed for this system. At exogenous substrate concentrations of 16 mM and a transmembrane pressure of 70 kPa, hexanal production rates are in the order of 5.1 microg/min. Addition of Triton X-100 resulted in membrane fouling and reduced flux. The reactor system has been run for periods of up to 1 week and has been shown to be stable over this period.

An active, thermostable cytochrome c reductant has been found in the cytosol of Phaseolus vulgaris cotyledons. This reducing activity is enhanced in the presence of NADH or NADPH and is attributable to a molecule of low molecular mass that is non-proteinaceous. The existence of this soluble reductant confounds measurements of enzymes that involve the use of exogenous electron acceptors and cofactors unless it is recognized and distinguished from true enzymatic activity.

Conventional wastewater disinfection is typically accomplished via chlorination, ozonation or ultraviolet (UV) irradiation. For each of these disinfectants, changes in the composition of the disinfected waters are known to occur and have been linked to changes in the toxicity responses of the disinfected effluent. The toxicity responses of wastewater effluent samples exposed to an alternative disinfectant, gamma (γ) radiation, have been studied and compared with the responses to chlorinated (with dechlorination) and undisinfected municipal wastewater effluent samples. Predisinfection samples were taken from six wastewater treatment plants. Multiple samples from each facility were split and subsamples were exposed to γ-irradiation, as well as chlorination/dechlorination in batch reactors. The disinfected and undisinfected subsamples were then assayed for toxicity using the Ceriodaphnia dubia chronic toxicity test. Wastewater disinfectant doses were sufficient to meet coliform limitations for effluent quality in all cases; furthermore, the batch disinfectant exposure protocols accomplished extents of coliform inactivation that were comparable to those observed in the disinfection unit operations at the full-scale facilities from which the samples were collected. Analysis of data from these assays showed that γ-irradiation induced significantly less toxicity in terms of C. dubia survival and reproduction than chlorination/dechlorination. Toxicity in the γ-irradiated effluent either decreased or was unchanged as compared to an undisinfected sample; effluent samples subjected to chlorination/dechlorination often showed a statistically significant increase in toxicity as compared to the undisinfected samples and the γ-irradiated samples.

Hexanal and cis-3-hexenal are principal flavor volatiles in ripe tomato fruit, but whether they accumulate during ripening or are formed upon maceration of the tissue has not been clarified. This has been addressed by measuring levels of these aldehydes in green and ripe fruit with discrimination between intrinsic aldehyde content and aldehyde generation following tissue disruption. Volatile sampling of tomato fruit homogenates was accomplished by purge/trapping, followed by thermal desorption on a gas chromatograph equipped with a mass selective detector. Incubation of some samples with alcohol dehydrogenase to convert the aldehydes to their respective alcohols permitted positive identification of the isomeric form of hexenal as cis-3-hex-enal. Red and green tomato fruit homogenized in buffer with saturated CaCl 2 contained low (0.1-0.8 μg g -1 fresh weight) levels of hexanal and cis-3-hexenal: thus there is minimal endogenous volatile content in intact fruit. Volatile levels increased rapidly, up to 10-fold, following homogenization of ripe tomato fruit in the absence of CaCl 2 , and more modestly in corresponding green tomato fruit homogenates. Incubation with the appropriate lipoxygenase/hydroperoxide lyase substrate (linoleic acid for hexanal, linolenic acid for cis-3-hexenal) doubled the amount of volatile compound produced. Hexanal generation was suppressed in the presence of linolenic acid, suggesting that the enzyme complex has greater affinity for this substrate. As well, levels of cis-3-hexenal, but not hexanal, tended to decline within 30 min of homogenization, possibly reflecting a specific degradative process. The results collectively indicate that the contribution of six-carbon aldehydes to tomato fruit flavor is attributable to metabolism invoked following tissue disruption rather than within the intact fruit.

The fate of ribulose-1,5-bisphosphate carboxylase was studied during development of carnation ( Dianthus cryophyllus L. cv. White Sim) petals. The petals of various developmental stages were dissected into upper wide and basal narrow parts, and analyzed separately. Rubisco activity was assessed in aqueous extracts of the petal parts in parallel with measurements of CO 2 fixation by intact petals. During development, CO 2 fixation declined markedly, Rubisco activity per corolla declined slightly and Rubisco activity per g dry weight increased. Analysis of the steady state level of the large and small subunits (LSu, SSu) of Rubisco by western blotting, demonstrated a decline in the amounts of both. The reduction in the level of SSu appeared prior to the decrease in the amount of LSu. An analysis of the spatial distribution of the Rubisco subunits revealed that the upper part of the petal was devoid of SSu in both young and old flowers. The data suggest that two different mechanisms are involved in the regulation of the decline in the amount of LSu. The first is at the transcriptional level and operates primarily in the basal part of the petal. The second mechanism operates in the upper part of the petal and may involve post-transcriptional processes.

The article questions the widely held view that nationalism was a significant feature of modem civilisation and particularly of nineteenth-century Europe. Two groups of events are chosen for examination: reputedly classic instances of nationalism (the French Revolution, German responses to Napoleon, the Italian and German revolutions of 1848, Italian and German unification and the Eastern crisis of 1875-8), and important international events which in an age of nationalism should reflect it (the Crimean War, Bismarck's alliances and the Franco-Russian alliance). Defined as the effort of nation/peoples to defend/extend their power, nationalism is evaluated specifically for its breadth of support and its influence on decision-makers which prove to be limited. This conclusion has implications not only for these events but also brings into question the established system of historical periodisation which presumes the distinctiveness of the nineteenth century and modem civilisation precisely because of their distinguishing features such as nationalism.

Six-carbon (C 6 ) volatile products are released from the enzymatic action of hydroperoxide lyase (HPL), a component of the lipoxygenase (LOX) pathway and form the basis of the green-note flavour characteristic of many consumed plant products. Arabidopsis leaf tissue contains the C 6 -aldehydes hexanal, and trans-2-hexenal as well as the C 6 -alcohols: hexanol, and 3-hexenol. Interconversion between C 6 -aldehydes and alcohols is thought to proceed through the action of alcohol dehydrogenase (ADH). Using an ADH mutant of Arabidopsis, we have shown that there are large quantitative and qualitative differences in the accumulation of C 6 -volatiles in the absence of ADH activity. The total quantity of LOX-derived volatiles is greater on a fresh weight basis in the ADH mutant. Qualitatively, hexanol and 3-hexenol levels are approximately 62% and 51% lower in the mutant, respectively, whereas levels of hexenal are approximately 10-fold higher. Hexanal accumulation, however, is unaffected in the mutant. The altered profile of LOX-derived volatiles does not have an effect on the steady-state levels of mRNA for allene oxide synthase (AOS) or LOX. HPL activity and mRNA quantity, however, are higher in the mutant relative to wild type, suggesting that altered product levels in the mutant affect HPL regulation.

Objective: To examine the utility of single-breath CO 2 analysis as a measure of lung volume. Design: A prospective, animal cohort study comparing 21 parameters derived from single-breath CO 2 analysis with lung volume measurements determined by nitrogen washout in animals during controlled ventilation. Setting: An animal laboratory in a university-affiliated medical center. Subjects: Seven healthy lambs. Interventions: The single-breath CO 2 analysis station consists of a mainstream capnometer, a variable orifice pneumotachometer, a signal processor and computer software with capability for both on- and off-line data analysis. Twenty-one derived components of the CO 2 expirogram were evaluated as predictors of lung volume. Lung volume was manipulated by 3 cm H 2 O incremental increases in positive end-expiratory pressure from 0 to 21 cm H 2 O, and ranged between 147 and 942 mL. Measurements and Main Results: Fifty-five measurements of lung volume were available for comparison with derived variables from the CO 2 expirogam. Stepwise linear regression identified four variables that were most predictive of lung volume: a) dynamic lung compliance; b) the slope of phase 3; c) the slope of phase 2 divided by the mixed expired CO 2 tension; and d) airway deadspace. The multivariate equation was highly statistically significant and explained 94% of the variance (adjusted r 2 =.94, p

Hydroperoxide lyase (HPL) cleaves lipid hydroperoxides to produce volatile flavor molecules and also potential signal molecules. We have characterized a gene from Arabidopsis that is homologous to a recently cloned HPL from green pepper (Capsicum annuum). The deduced protein sequence indicates that this gene encodes a cytochrome P-450 with a structure similar to that of allene oxide synthase. The gene was cloned into an expression vector and expressed in Escherichia coli to demonstrate HPL activity. Significant HPL activity was evident when 13S-hydroperoxy-9(Z),11(E),15(Z)-octadecatrienoic acid was used as the substrate, whereas activity with 13S-hydroperoxy-9(Z),11(E)-octadecadienoic acid was approximately 10-fold lower. Analysis of headspace volatiles by gas chromatography-mass spectrometry, after addition of the substrate to E. coli extracts expressing the protein, confirmed enzyme-activity data, since cis-3-hexenal was produced by the enzymatic activity of the encoded protein, whereas hexanal production was limited. Molecular characterization of this gene indicates that it is expressed at high levels in floral tissue and is wound inducible but, unlike allene oxide synthase, it is not induced by treatment with methyl jasmonate.

Lipid-protein particles bearing the 55-kD ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) (EC 4.1.1.39) large subunit (RLSU) and no detectable corresponding Rubisco small subunit (RSSU) were isolated from the stroma of intact chloroplasts by flotation centrifugation. Stromal RLSU-bearing particles appear to originate from thylakoids because they can also be generated in vitro by illumination of isolated thylakoids. Their formation in vitro is largely heat denaturable and is facilitated by light or ATP. RLSU-containing lipid-protein particles range from 0.05 to 0.10 [mu]m in radius, contain the same fatty acids as thylakoids, but have a 10- to 15-fold higher free-to-esterified fatty acid ratio than thylakoids. RLSU-bearing lipid-protein particles with no detectable RSSU were also immunopurified from the populations of both stromal lipid-protein particles and those generated in vitro from illuminated thylakoids. Protease shaving indicated that the RLSU is embedded in the lipid-protein particles and that there is also a protease-protected RLSU in thylakoids. These observations collectively indicate that the RLSU associated with thylakoids is released into the stroma by light-facilitated blebbing of lipid-protein particles. The release of RLSU-containing particles may in turn be coordinated with the assembly of Rubisco holoenzyme because chaperonin 60 is also associated with lipid-protein particles isolated from stroma.

Objective: To compare bag-mask ventilation performed by emergency department (ED) personnel using anesthesia bags (AB) and self-inflating bags (SIB). Setting: ED in a teaching children's hospital where the AB is the device used during resuscitations. Design: Experimental study. Bag-mask ventilation was evaluated with an infant resuscitation mannequin equipped to measure airway volumes and pressures. Pediatric residents, ED nurses, and pediatric emergency medicine fellows performed bag-mask ventilation with AB and SIB and rated their confidence using each device. Main outcome measure: Ventilation failure rates. Results: Seventy subjects participated (17 interns, 16 junior residents, 13 senior residents, 10 fellows, and 14 nurses). There were 13 failures with the AB (18.6%) versus 1 (1.4%) with the SIB (P < 0.01) [95% confidence interval: 5-29%], with a significant difference even after excluding the least experienced subjects. There was no difference in high pressure breaths delivered (SIB 19% vs AB 15%, P = 0.4) and a higher incidence of hyperventilation with the SIB (67 vs 25%, P < 0.01). While using the SIB, 19 (27%) of the subjects did not turn on the O 2 flow. There was no difference in pretest confidence rating, but the posttest confidence rating was higher for the SIB (P < 0.05). Conclusions: Compared to SIB use for bag-mask ventilation in an ED, AB use resulted in more ventilation failures, no advantage in preventing excessive airway pressures, and less confidence among operators. The SIB should be the first choice for bag-mask ventilation in the ED, with attention to maximize oxygen delivery.