bluuebunny/crossref_metadata_2025 huggingface dataset:
Quantitative Proteomics of Seed Filling in Castor: Comparison with Soybean and Rapeseed Reveals Differences between Photosynthetic and Nonphotosynthetic Seed Metabolism
Authors: Houston Norma L. , Hajduch Martin , Thelen Jay J.
Published: August 2009

Seed maturation or seed filling is a phase of development that plays a major role in the storage reserve composition of a seed. In many plant seeds photosynthesis plays a major role in this process, although oilseeds, such as castor (Ricinus communis), are capable of accumulating oil without the benefit of photophosphorylation to augment energy demands. To characterize seed filling in castor, a systematic quantitative proteomics study was performed. Two-dimensional gel electrophoresis was used to resolve and quantify Cy-dye-labeled proteins expressed at 2, 3, 4, 5, and 6 weeks after flowering in biological triplicate. Expression profiles for 660 protein spot groups were established, and of these, 522 proteins were confidently identified by liquid chromatography-tandem mass spectrometry by mining against the castor genome. Identified proteins were classified according to function, and the most abundant groups of proteins were involved in protein destination and storage (34%), energy (19%), and metabolism (15%). Carbon assimilatory pathways in castor were compared with previous studies of photosynthetic oilseeds, soybean (Glycine max) and rapeseed (Brassica napus). These comparisons revealed differences in abundance and number of protein isoforms at numerous steps in glycolysis. One such difference was the number of enolase isoforms and their sum abundance; castor had approximately six times as many isoforms as soy and rapeseed. Furthermore, Rubisco was 11-fold less prominent in castor compared to rapeseed. These and other differences suggest some aspects of carbon flow, carbon recapture, as well as ATP and NADPH production in castor differs from photosynthetic oilseeds.
Remodeled Respiration in ndufs4 with Low Phosphorylation Efficiency Suppresses Arabidopsis Germination and Growth and Alters Control of Metabolism at Night
Authors: Meyer Etienne H. , Tomaz Tiago , Carroll Adam J. , Estavillo Gonzalo , Delannoy Etienne , Tanz Sandra K. , Small Ian D. , Pogson Barry J. , Millar A. Harvey
Published: August 2009

Respiratory oxidative phosphorylation is a cornerstone of cellular metabolism in aerobic multicellular organisms. The efficiency of this process is generally assumed to be maximized, but the presence of dynamically regulated nonphosphorylating bypasses implies that plants can alter phosphorylation efficiency and can benefit from lowered energy generation during respiration under certain conditions. We characterized an Arabidopsis (Arabidopsis thaliana) mutant, ndufs4 (for NADH dehydrogenase [ubiquinone] fragment S subunit 4), lacking complex I of the respiratory chain, which has constitutively lowered phosphorylation efficiency. Through analysis of the changes to mitochondrial function as well as whole cell transcripts and metabolites, we provide insights into how cellular metabolism flexibly adapts to reduced phosphorylation efficiency and why this state may benefit the plant by providing moderate stress tolerance. We show that removal of the single protein subunit NDUFS4 prevents assembly of complex I and removes its function from mitochondria without pleiotropic effects on other respiratory components. However, the lack of complex I promotes broad changes in the nuclear transcriptome governing growth and photosynthetic function. We observed increases in organic acid and amino acid pools in the mutant, especially at night, concomitant with alteration of the adenylate content. While germination is delayed, this can be rescued by application of gibberellic acid, and root growth assays of seedlings show enhanced tolerance to cold, mild salt, and osmotic stress. We discuss these observations in the light of recent data on the knockout of nonphosphorylating respiratory bypass enzymes that show opposite changes in metabolites and stress sensitivity. Our data suggest that the absence of complex I alters the adenylate control of cellular metabolism.
Protective effects of affinity‐purified antibody and truncated vaccines against Pseudomonas aeruginosa V‐antigen in neutropenic mice
Authors: Moriyama Kiyoshi , Wiener‐Kronish Jeanine P. , Sawa Teiji
Published: August 2009

Virulent P. aeruginosa strains express PcrV, one of the translocational components of the type III secretion system. PcrV has been reported to be a protective antigen against lethal P. aeruginosa infection. The PcrV region, which contributes to protective immunity against P. aeruginosa infection, was investigated by using genetically engineered, truncated PcrV proteins and affinity‐purified anti‐PcrV antibodies against the truncated PcrV proteins. The efficacy of active and passive immunization against PcrV was tested in mice with cyclophosphamide‐induced immunosuppression by intraabdominal challenge of P. aeruginosa. Active immunization with either full‐length PcrV1‐294 or PcrV139‐294 significantly improved the survival of mice infected with P. aeruginosa, while PcrV139‐258, PcrV139‐234, PcrV197‐294, and PcrV261‐294 were not protective. These results suggest that an effective PcrV vaccine needs to contain not only the Mab166 epitope (PcrV144‐257) but also the carboxyl terminal tail of PcrV. In the case of passive immunization, administration of affinity‐purified anti‐PcrV IgG against either PcrV1‐294 or PcrV139‐258 showed significantly higher efficacy against lethal P. aeruginosa infection than did original anti‐PcrV IgG and Mab166. The increased efficacy of affinity‐purified anti‐PcrV IgG implies that more potent anti‐PcrV strategies are possible. The results of this study are crucial to the development of an effective PcrV vaccine for active immunization and to an appropriate blocking anti‐PcrV antibody against P. aeruginosa infection in humans.
Anti‐myeloma activity of the novel 2‐aminothienopyrimidine Hsp90 inhibitor NVP‐BEP800
Authors: Stühmer Thorsten , Chatterjee Manik , Grella Evelyn , Seggewiss Ruth , Langer Christian , Müller Sabine , Schoepfer Joseph , Garcia‐Echeverria Carlos , Quadt Cornelia , Jensen Michael R. , Einsele Hermann , Bargou Ralf C.
Published: August 2009

The 90 kD heat shock protein (Hsp90) molecular chaperone sustains multiple components of oncogenic pathways and has recently emerged as a therapeutic target that is now being clinically tested in a number of malignancies. In order to address formulation issues and to deal with possible resistance mechanisms against small molecule Hsp90 inhibitors, a range of compounds based on different molecular scaffolds are now being developed. The present study preclinically tested the effects of the novel 2‐aminothienopyrimidine class Hsp90 inhibitor NVP‐BEP800, which is suitable for oral formulations, on multiple myeloma cells from established cell lines and on a larger cohort (n = 40) of primary myeloma samples. The drug effectively and specifically killed the majority of primary myeloma cells in coculture with bone marrow stromal cells and reliably entailed molecular consequences of Hsp90 blockade – such as survival pathway breakdown and client protein depletion – in multiple myeloma cells from cell lines as well as from patients. Collectively, the properties of this novel drug support clinical testing in multiple myeloma.
A morphological classification capturing functional variation in phytoplankton
Authors: KRUK CARLA , HUSZAR VERA L. M. , PEETERS EDWIN T. H. M. , BONILLA SYLVIA , COSTA LUCIANA , LÜRLING MIQUEL , REYNOLDS COLIN S. , SCHEFFER MARTEN
Published: August 2009

1. A logical way of distinguishing functional groups of phytoplankton is to cluster species according to their functional traits, such as growth rate and nutrient assimilation constants. However, data for such an approach are lacking for the vast majority of the species.

2. In this study, we show that a classification based on simple morphological traits may capture much of the variability in functional properties among the phytoplankton. We used information on more than 700 freshwater species, from more than 200 lakes situated in climate zones ranging from subpolar to tropical.

3. Morphological characteristics correlated well with functional properties, such as growth rate and sinking rate, and also with the population size and biomass attained in the field. This suggests that morphology is a good predictor of the functional characteristics of species.

4. Cluster analysis was used to define seven species groups based on morphology. Although some of the clusters are taxonomically homogeneous, others include species of several separate divisions. Functional traits (not used for the classification) differed significantly among the clusters, suggesting that the clusters may indeed represent meaningful functional groups.

5. Advantages of our morphological approach to classification include its objectivity, its independence from taxonomic affiliations, and the relative ease of its application to the majority of species for which physiological traits are unknown and are not readily determined.
Essential requirement for RING finger E3 ubiquitin ligase Hakai in early embryonic development of Drosophila
Authors: Kaido Masako , Wada Housei , Shindo Masayo , Hayashi Shigeo
Published: August 2009

Hakai is a RING finger type E3 ubiquitin ligase that is highly conserved in metazoans. Mammalian Hakai was shown to bind and ubiquitinate the intracellular domain of E‐cadherin, and this activity is implicated in down‐regulation of E‐cadherin during v‐Src‐induced cellular transformation. To evaluate this model in vivo, we studied the function of the Drosophila homologue of Hakai. In cultured S2 cells, Drosophila Hakai and E‐cadherin (Shotgun) formed a complex in a way distinct from the interaction described for mammalian counterparts. Hakai null mutants died during larval stages but this lethality could be offset by a HA‐tagged Hakai construct. While zygotic Hakai function was dispensable for cell proliferation and differentiation in the wing disc epithelium, maternal Hakai mutants showed a variety of defects in epithelial integrity, including stochastic loss of E‐cadherin expression and reduction of aPKC; defects in cell specification and cell migration were also observed. No increase of E‐cadherin, however, was observed. Regulation of multiple target proteins under control of Hakai is, therefore, essential for early embryonic morphogenesis in Drosophila.
Fission yeast Ku protein is required for recovery from DNA replication stress
Authors: Miyoshi Tomoichiro , Kanoh Junko , Ishikawa Fuyuki
Published: August 2009

The fundamental function of the conserved Ku70–Ku80 heterodimer is to promote the non‐homologous end‐joining (NHEJ) pathway in double‐strand break repair. Although it is thought that Ku plays several roles other than NHEJ in maintaining chromosomal integrity including telomere protection, these precise functions remain unclear. In this study, we describe a novel role of fission yeast Ku proteins encoded by pku70+ and pku80+ genes in dealing with DNA replication stress. In the absence of Rqh1, the fission yeast RecQ helicase, the cells are sensitive to reagents inducing replication stress. pkuΔrqh1Δ double mutant showed synergistic sensitivities to these reagents. However, this synthetic phenotype was not observed when rqh1Δ mutant was coupled with the deletion of lig4+ that encodes a ligase essential for NHEJ, indicating that the role of Ku in replication stress is NHEJ independent. pkuΔrqh1Δ double mutant also showed highly variable copy numbers of rDNA repeats even under unstressed condition. Furthermore, the double mutant exhibited inefficient replication resumption after transient replication stalling. These results suggest the possibility that Ku proteins play an important role in genome integrity recovering replication stress.
Induction of hyperproliferative fetal megakaryopoiesis by an N‐terminally truncated GATA1 mutant
Authors: Shimizu Ritsuko , Kobayashi Eri , Engel James Douglas , Yamamoto Masayuki
Published: August 2009

Two GATA1‐related leukemias have been described: one is an erythroleukemia that develops in mice as a consequence of diminished expression of wild‐type GATA1, whereas the other is an acute megakaryoblastic leukemia (AMKL) that arises in Down syndrome children as a consequence of somatic N‐terminal truncation (ΔNT) of GATA1. We discovered that mice expressing the shortened GATA1 protein (ΔNTR mice) phenocopies the human transient myeloproliferative disorder (TMD) that precedes AMKL in Down syndrome children. In perinatal livers of the ΔNTR mutant mice, immature megakaryocytes accumulate massively, and this fraction contains cells that form hyperproliferative megakaryocytic colonies. Furthermore, showing good agreement with the clinical course of TMD in humans, ΔNTR mutant mice undergo spontaneous resolution from the massive megakaryocyte accumulation concomitant with the switch of hematopoietic microenvironment from liver to bone marrow/spleen. These results thus demonstrate that expression of the GATA1/Gata1 N‐terminal deletion mutant per se induces hyperproliferative fetal megakaryopoiesis. This mouse model serves as an important means to clarify how impaired GATA1 function contributes to the multi‐step leukemogenesis.
Prolyl isomerase Pin1 shares functional similarity with phosphorylated CTD interacting factor PCIF1 in vertebrate cells
Authors: Yunokuchi Izumi , Fan Hong , Iwamoto Yu , Araki Chisato , Yuda Masamichi , Umemura Hiroyasu , Harada Fumio , Ohkuma Yoshiaki , Hirose Yutaka
Published: August 2009

The carboxy‐terminal domain (CTD) of the RNA polymerase II (Pol II) largest subunit undergoes reversible phosphorylation during transcription cycle. The phosphorylated CTD plays critical roles in coordinating transcription with chromatin modification and RNA processing by serving as a scaffold to recruit various proteins. Recently, we identified a novel human WW domain‐containing protein PCIF1 as a phosphorylated CTD‐interacting factor and demonstrated that PCIF1 negatively modulates Pol II activity in vivo. In the present study, to explore cellular functions of PCIF1, we generated PCIF1‐deficient chicken DT40 cell lines. We observed significant up‐regulation of WW domain‐containing prolyl isomerase Pin1 in two independently established PCIF1‐deficient mutant clones. As reconstitution of PCIF1 in the mutants did not reduce Pin1 expression, PCIF1 may not be a negative regulator of Pin1 expression. We assume that Pin1 over‐expression might suppress defects caused by PCIF1 deficiency in DT40 cells. We furthermore compared PCIF1 and Pin1 for their functional properties and found that these two proteins exhibit most similar target specificity among other CTD‐binding WW proteins, overlapping subcellular localization and comparative inhibitory effects on transcriptional activation by Pol II in human cultured cells. These results suggest that Pin1 may have overlapping cellular function with PCIF1 in vertebrate cells.
The role of electronic pocket dictionaries as an English learning tool among Chinese students
Authors: Jian Hua‐Li , Sandnes Frode Eika , Law Kris M. Y. , Huang Yo‐Ping , Huang Yueh‐Min
Published: August 2009

This study addressed the role of electronic pocket dictionaries as a language learning tool among university students in Hong Kong and Taiwan. The target groups included engineering and humanities students at both undergraduate and graduate level. Speed of reference was found to be the main motivator for using an electronic pocket dictionary. Next, the functionality used was found to be connected to the language proficiency of the learner. Finally, multimedia content was ranked least important. The results of this study have implications for the design of electronic dictionaries and for the teaching of second languages with electronic dictionaries. In particular, device developers should focus on improving the accessing speed and pay less attention to multimedia functionality. Educators should ensure that the device functionality matches the language proficiency level of the students.
Factor structure of the Athens Insomnia Scale and its associations with demographic characteristics and depression in adolescents
Authors: YEN CHENG‐FANG , KING BRYAN H. , CHANG YU‐PING
Published: August 2009

The eight‐item Athens Insomnia Scale (AIS‐8) is an instrument that has been used frequently to assess insomnia problems. Previous research on adults has found that the AIS‐8 functioned as a sole component. This study aimed to examine the prevalence rates of insomnia problems on the AIS‐8, the factor structure of the AIS‐8 in adolescents and its associations with demographic characteristics and depression in adolescents. A total of 8319 adolescent students (4334 girls and 3985 boys, mean age = 14.7 years, standard deviation = 1.7 years) in southern Taiwan were recruited into this study and completed the AIS‐8. We performed an exploratory factor analysis to examine the factor structure of the AIS‐8, and used the parallel analysis for making decisions regarding factor retention. We also used multiple regression analysis models to determine the associations between insomnia and demographic characteristics and depression. The results found that a high proportion of adolescents had insomnia problems as measured by the AIS‐8. The AIS‐8 was composed of two different factors when used among a large adolescent population, including insomnia symptoms (factor 1) and subjective sleep and daytime distress (factor 2). While being male, being younger, and having depression were associated positively with the severity of insomnia symptoms (factor 1), being older, living in urban areas, and having depression were associated positively with the severity of subjective sleep and daytime distress (factor 2). Clinicians and researchers should consider the different meanings of the two factors of the AIS‐8 when using this tool to assess insomnia problems in adolescents.
Sleep state distribution of obstructive events in children: is obstructive sleep apnoea really a rapid eye movement sleep‐related condition?
Authors: VERGINIS NICOLE , JOLLEY DAMIEN , HORNE ROSEMARY S. C. , DAVEY MARGOT J. , NIXON GILLIAN M.
Published: August 2009

Obstructive sleep apnoea (OSA) in children is commonly considered to occur predominantly in rapid eye movement (REM) sleep, but clinical experience suggests that this is not universally the case. We hypothesized that there would be a subgroup of children with OSA who have non‐REM (NREM) predominance of obstructive events and that these children share certain clinical characteristics. Thus, we aimed to compare the obstructive apnoea–hypopnoea index (OAHI) in REM versus NREM sleep and to assess factors influencing the distribution of events by sleep state. Polysomnography (PSG) recordings of 102 children aged 0–18 years with moderate to severe OSA (OAHI ≥5 h−1) were reviewed. OAHI was calculated separately for REM and NREM sleep. A REM predominance index (RPI) was determined using log transformation [RPI = log (REM OAHI + 0.5) − log (NREM OAHI + 0.5)] and compared with possible influencing factors using multiple linear regression. Analysis showed that obstructive events were more common in REM sleep (median REM OAHI 21.4 h−1, median NREM OAHI 8.3 h−1, P < 0.001). Mean RPI was significantly greater than zero (P = 0.003). However, a substantial minority of children (30.4%) had a higher NREM than REM OAHI. The factors that were related significantly to NREM predominance were older age (P = 0.02), higher arousal index (P < 0.001) and higher SpO2 nadir (P < 0.001). Our findings demonstrate that while OSA is a REM sleep‐related problem in the majority of children, there is a significant subset of children with NREM predominance of obstructive events. This finding highlights the importance of considering sleep state distribution of events in studies of the pathophysiology and outcomes of OSA in childhood.
Association study of IFNAR2 and IL10RB genes with the susceptibility and interferon response in HBV infection
Authors: Gong Q.‐M. , Kong X.‐F. , Yang Z.‐T. , Xu J. , Wang L. , Li X.‐H. , Jin G.‐D. , Gao J. , Zhang D.‐H. , Jiang J.‐H. , Lu Z.‐M. , Zhang X.‐X.
Published: August 2009

Summary.  A recent genome‐wide association study discovered that two polymorphisms, interferon (IFN) alpha receptor 2 (IFNAR‐2) F8S and interleukin 10 receptor (IL10RB) K47E, were associated with susceptibility to hepatitis B virus (HBV) infection in Africa. Here, we reevaluate the effects of the two polymorphisms on HBV susceptibility in the Chinese Han population, and extended the study to look at their association with IFN response in chronic hepatitis B (CHB). We included 341 patients with CHB and 341 unrelated controls presenting with asymptotic HBV self‐limited infection, who were well matched in age and sex. In the CHB group, 101 patients had been treated with peg‐IFN‐alpha‐2a for 48 weeks and followed up for 24 weeks to determine the clinical response, resulting 34 individuals with sustained virological response (SVR) and 67 individuals with nonsustained response (NR). Subgroups in the CHB group were divided according to the viral loads, HBeAg and maternal HBsAg status. The association with the susceptibility to HBV infection was only observed for IL10RB K47E when we compared the individuals with persistent HBV infection through nonmaternal transmission to the controls with asymptomatic self‐limited HBV infection. Further, we found that the IFNAR2‐8SS genotype was associated with HBeAg negative patients (OR = 0.316, 95% CI: 0.121–0.825, P = 0.019) and that the IFNAR2‐8F allele was associated with the risk to high viral loads (OR = 1.667, 95% CI: 1.148–2.420, P = 0.007). In addition, the IFNAR2‐8FF genotype predisposed to higher MxA gene induction and correlated with sustained IFN response (OR = 0.348, 95% CI: 0.129–0.935, P = 0.036). Haplotype analysis based on polymorphisms of three single‐nucleotide polymorphisms, MxA −88 G/T, IFNAR‐2 F8S and IL10RB K47E showed that the haplotype distribution was significantly different between the SVR and NR groups (P = 0.040). This study suggests that IFNAR2 may play an important role in determining IFN response and clinical phenotypes of HBV infection in the Chinese Han population.
Switching off small RNA regulation with trap‐mRNA
Authors: Overgaard Martin , Johansen Jesper , Møller‐Jensen Jakob , Valentin‐Hansen Poul
Published: August 2009

Small non‐coding regulatory RNAs in bacteria have been shown predominantly to be tightly regulated at the level of transcription initiation, and sRNAs that function by an antisense mechanism on trans‐encoded target mRNAs have been shown or predicted to act stoichiometrically. Here we show that MicM, which silences the expression of an outer membrane protein, YbfM under most growth conditions, does not become destabilized by target mRNA overexpression, indicating that the small RNA regulator acts catalytically. Furthermore, our regulatory studies suggested that control of micM expression is unlikely to operate at the level of transcription initiation. By employing a highly sensitive genetic screen we uncovered a novel RNA‐based regulatory principle in which induction of a trap‐mRNA leads to selective degradation of a small regulatory RNA molecule, thereby abolishing the sRNA‐based silencing of its cognate target mRNA. In the present case, antisense regulation by chb mRNA of the antisense regulator MicM by an extended complementary sequence element, results in induction of ybfM mRNA translation. This type of regulation is reminiscent of the regulation of microRNA activity through target mimicry that occurs in plants.
Targeted σ factor turnover inserts negative control into a positive feedback loop
Authors: Donohue Timothy J.
Published: August 2009

Since their classification as members of the σ70 superfamily, Group IV alternative σ factors have been found to control gene expression in response to diverse environmental or stress signals. Activity of the Streptomyces coelicolor Group IV family member, σR (SigR), is increased by changes in the oxidation‐reduction state of cytoplasmic disulphide bonds. Once released by its cognate anti‐σ factor RsrA, σR activates expression of gene products that help cells reduce cytoplasmic disulphide bonds. In this issue of Molecular Microbiology, Kim and co‐workers provide new insights into positive and negative control of σR activity. The authors show that a transcript derived from the inducible σR‐dependent sigRrsrA p2 promoter operon encodes a σR protein of a higher molecular weight (termed σR′) than is found in uninduced cells. One major difference between σR′ and the smaller σR protein found in uninduced cells is the rapid proteolysis of σR′ by the ClpP1/P2 protease system. The genes for the ClpP1/ClpP2 protease subunits are themselves members of the σR regulon. The newly identified positive (σR′ synthesis) and negative control (selective σR′ turnover) aspects of this circuit are either found or predicted to exist in other related Group IV σ factor family members.
Mucosal mast cells mediate motor response induced by chronic oral exposure to ovalbumin in the rat gastrointestinal tract
Authors: Traver E. , Torres R. , De Mora F. , Vergara P.
Published: August 2009

Abstract  We previously demonstrated that oral chronic exposure to ovalbumin (OVA) causes intestinal hypermotility in Sprague‐Dawley rats. In this study, the objective was to determine the mechanism of action of OVA and the role of mucosal mast cells in the regulation of motor activity in this model. Rats were orally exposed to OVA during 6 weeks. Intestinal mucosal mast cells (IMMCs) were counted and rat mast cell protease II (RMCPII) measured in duodenum, jejunum, ileum and colon. Anti‐OVA IgE, IgG, and IL‐4 were measured in serum. Eosinophils and IgE+ cells were counted in jejunum. In an additional study rats were treated with the mast cell stabilizer ketotifen and mast cell number, RMCPII concentration and motor activity in vitro were evaluated. OVA exposed rats showed an increase in mucosal mast cell number and in RMCPII content in small intestine and colon. However, variables of a Th2 type response were not affected by exposure to OVA: (i) neither OVA specific IgE nor IgG were found; (ii) IL‐4 did not increase and, (iii) the number of eosinophils and IgE+ cells was identical in the exposed and unexposed groups. These results brought us to hypothesize a possible non‐Ig‐mediated action of OVA on mast cells. Ketotifen significantly diminished the response to OVA: Ketotifen reduced the number of mast cells and the RMCPII content and blocked increased intestinal contractility. In addition ketotifen modified motor response in both OVA exposed and unexposed animals giving evidence of the importance of mast cells in intestine motor activity driving.
Tissue and cellular localization of individual β‐glycosidases using a substrate‐specific sugar reducing assay
Authors: Sánchez‐Pérez Raquel , Jørgensen Kirsten , Motawia Mohammed S. , Dicenta Federico , Møller Birger L.
Published: August 2009

Traditional methods to localize β‐glycosidase activity in tissue sections have been based on incubation with the general substrate 6‐bromo‐2‐naphthyl‐β‐d‐glucopyranoside. When hydrolysed in the presence of salt zinc compounds, 6‐bromo‐2‐naphthyl‐β‐d‐glucopyranoside affords the formation of an insoluble coloured product. This technique does not distinguish between different β‐glycosidases present in the tissue. To be able to monitor the occurrence of individual β‐glycosidases in different tissues and cell types, we have developed a versatile histochemical method that can be used for localization of any β‐glycosidase that upon incubation with its specific substrate releases a reducing sugar. Experimentally, the method is based on hydrolysis of the specific substrate followed by oxidation of the sugar released by a tetrazolium salt (2,3,5‐triphenyltetrazolium chloride) that forms a red insoluble product when reduced. The applicability of the method was demonstrated by tissue and cellular localization of two β‐glucosidases, amygdalin hydrolase and prunasin hydrolase, in different tissues and cell types of almond. In those cases where the analysed tissue had a high content of reducing sugars, this resulted in strong staining of the background. This interfering staining of the background was avoided by prior incubation with sodium borohydride. The specificity of the devised method was demonstrated in a parallel localization study using a specific antibody towards prunasin hydrolase.
FATTY ACID DESATURASE4 of Arabidopsis encodes a protein distinct from characterized fatty acid desaturases
Authors: Gao Jinpeng , Ajjawi Imad , Manoli Arthur , Sawin Andrew , Xu Changcheng , Froehlich John E. , Last Robert L. , Benning Christoph
Published: August 2009

Polar membrane glycerolipids occur in a mixture of molecular species defined by a polar head group and characteristic acyl groups esterified to a glycerol backbone. A molecular species of phosphatidylglycerol specific to chloroplasts of plants carries a Δ3‐trans hexadecenoic acid in the sn‐2 position of its core glyceryl moiety. The fad4‐1 mutant of Arabidopsis thaliana missing this particular phosphatidylglycerol molecular species lacks the necessary fatty acid desaturase, or a component thereof. The overwhelming majority of acyl groups associated with membrane lipids in plants contains double bonds with a cis configuration. However, FAD4 is unusual because it is involved in the formation of a trans double bond introduced close to the carboxyl group of palmitic acid, which is specifically esterified to the sn‐2 glyceryl carbon of phosphatidylglycerol. As a first step towards the analysis of this unusual desaturase reaction, the FAD4 gene was identified by mapping of the FAD4 locus and coexpression analysis with known lipid genes. FAD4 encodes a predicted integral membrane protein that appears to be unrelated to classic membrane bound fatty acid desaturases based on overall sequence conservation. However, the FAD4 protein contains two histidine motifs resembling those of metalloproteins such as fatty acid desaturases. FAD4 is targeted to the plastid. Overexpression of the cDNA in transgenic Arabidopsis led to increased accumulation of the Δ3‐trans hexadecanoyl group in phosphatidylglycerol relative to wild type. Taken together these results are consistent with the hypothesis that FAD4 is the founding member of a novel class of fatty acid desaturases.
Automated pre‐ejection period variation predicts fluid responsiveness in low tidal volume ventilated pigs
Authors: VISTISEN S. T. , KOEFOED‐NIELSEN J. , LARSSON A.
Published: August 2009

Introduction: The respiratory variation in the pre‐ejection period (ΔPEP) has been used to predict fluid responsiveness in mechanically ventilated patients. Recently, we automated this parameter and indexed it to tidal volume (PEPV) and showed that it was a reliable predictor for post‐cardiac surgery, mainly paced, patients ventilated with low tidal volumes. The aims of the present animal study were to investigate PEPV's ability to predict fluid responsiveness under different fluid loading conditions and natural heart rates during low tidal volume ventilation (6 ml/kg) and to compare the performance of PEPV with other markers of fluid responsiveness.

Methods: Eight prone, anesthetized piglets (23–27 kg) ventilated with tidal volumes of 6 ml/kg were subjected to a sequence of 25% hypovolemia, normovolemia, and 25% and 50% hypervolemia. PEPV, ΔPEP, pulse pressure variation (PPV), central venous pressure (CVP), and pulmonary artery occlusion pressure (PAOP) were measured before each volume expansion.

Results: Sensitivity was 89% and specificity was 93% for PEPV, 78% and 93% for ΔPEP, 89% and 100% for PPV, 78% and 93% for CVP, and 89% and 87% for PAOP.

Conclusion: PEPV predicts fluid responsiveness in low tidal volume ventilated piglets.
Late sensory function after intraoperative capsaicin wound instillation
Authors: AASVANG E. K. , HANSEN J. B. , KEHLET H.
Published: August 2009

Background: Intense capsaicin‐induced C‐fiber stimulation results in reversible lysis of the nerve soma, thereby making capsaicin wound instillation of potential interest for the treatment of post‐operative pain. Clinical histological and short‐term sensory studies suggest that the C‐fiber function is partly re‐established after skin injection of capsaicin. However, no study has evaluated the long‐term effects of wound instillation of purified capsaicin on sensory functions.

Methods: Patients included in a double‐blind placebo‐controlled randomized study of the analgesic effect of capsaicin after groin hernia repair were examined by quantitative sensory testing before, 1 week and 2 years post‐operatively. The primary endpoint was occurrence of hyperalgesia/allodynia. The secondary endpoints were acute and late sensory changes between the two patient groups. Patients were blinded to the allocated treatment.

Results: Twenty (100%) capsaicin and 16 (76%) placebo‐treated patients were seen at the year follow‐up. Hyperalgesia was seen in five capsaicin‐ vs. one placebo‐treated patient (P=0.2). The mechanical detection threshold was significantly increased on the operated side in the capsaicin vs. placebo group at the 1‐week follow‐up (P<0.05), but was not different at the year follow‐up (P=0.3). There were no other significant differences in sensory function on the operated side between groups at the pre‐operative, 1‐week or year post‐operative follow‐up (P>0.05). The sensory function on the contralateral side was comparable between groups throughout the study (P>0.1).

Conclusion: This small‐volume study calls for further long‐term safety studies of wound capsaicin instillation.
Visualization of alveolar recruitment in a porcine model of unilateral lung lavage using 3He‐MRI
Authors: RUDOLPH A. , MARKSTALLER K. , GAST K. K. , DAVID M. , SCHREIBER W. G. , EBERLE B.
Published: August 2009

Background: In the acute respiratory distress syndrome potentially recruitable lung volume is currently discussed. 3He‐magnetic resonance imaging (3He‐MRI) offers the possibility to visualize alveolar recruitment directly.

Methods: With the approval of the state animal care committee, unilateral lung damage was induced in seven anesthetized pigs by saline lavage of the right lungs. The left lung served as an intraindividual control (healthy lung). Unilateral lung damage was confirmed by conventional proton MRI and spiral‐CT scanning. The total aerated lung volume was determined both at a positive end‐expiratory pressure (PEEP) of 0 and 10 mbar from three‐dimensionally reconstructed 3He images, both for healthy and damaged lungs. The fractional increase of aerated volume in damaged and healthy lungs, followed by a PEEP increase from 0 to 10 mbar, was compared.

Results: Aerated gas space was visualized with a high spatial resolution in the three‐dimensionally reconstructed 3He‐MR images, and aeration defects in the lavaged lung matched the regional distribution of atelectasis in proton MRI. After recruitment and PEEP increase, the aerated volume increased significantly both in healthy lungs from 415 ml [270–445] (median [min–max]) to 481 ml [347–523] and in lavaged lungs from 264 ml [71–424] to 424 ml [129–520]. The fractional increase in lavaged lungs was significantly larger than that in healthy lungs (healthy: 17% [11–38] vs. lavage: 42% [14–90] (P=0.031).

Conclusion: The 3He‐MRI signal might offer an experimental approach to discriminate atelectatic vs. poor aerated lung areas in a lung damage animal model. Our results confirm the presence of potential recruitable lung volume by either alveolar collapse or alveolar flooding, in accordance with previous reports by computed tomography.
Levosimendan cardioprotection in acutely β‐1 adrenergic receptor blocked open chest pigs
Authors: METZSCH C. , LINNÉR R. , STEEN S. , LIAO Q. , ALGOTSSON L.
Published: August 2009

Background: Levosimendan and volatile anesthetics have myocardial pre‐conditioning effects. β‐1 adrenergic receptor antagonists may inhibit the protective effect of volatile anesthetics. No information exists as to whether this also applies to the pre‐conditioning effect of levosimendan. We therefore investigated whether levosimendan added to metoprolol would demonstrate a cardioprotective effect.

Methods: Three groups of anesthetized open chest pigs underwent 30 min of myocardial ischemia and 90 min of reperfusion by temporary occlusion of the largest side branch from the circumflex artery or the left anterior descending artery. One group (CTRL) served as a control, in another group (BETA), a metoprolol‐loading dose was intravenously injected 30 min before ischemia, and in a third group (BETA+L), a levosimendan infusion was added to metoprolol. Myocardial tissue concentrations of glucose, glycerol, and lactate/pyruvate ratio as the primary end‐points were investigated with microdialysis in ischemic and non‐ischemic tissues.

Results: At the end of the ischemic period, statistically significant differences were only found between CTRL and BETA+L in the ischemic myocardium, with a lower lactate/pyruvate ratio, lower glycerol, and higher glucose concentrations in BETA+L as compared with CTRL. There were no differences in non‐ischemic myocardium. From 10 to 90 min of reperfusion, no more differences were found between groups.

Conclusion: The cardioprotective effect of levosimendan on ischemic metabolism with a reduction in the myocardial lactate/pyruvate ratio, less glycerol accumulation, and better preserved glucose concentration does not seem to be prevented by β‐1 adrenergic receptor antagonism with metoprolol.
Improved outcome after trauma care in university‐level intensive care units
Authors: ALA‐KOKKO T. I. , OHTONEN P. , KOSKENKARI J. , LAURILA J. J.
Published: August 2009

Background: Centralized trauma care has been shown to be associated with improved patient outcome. We compared the outcomes of trauma patients in relation to the size of the intensive care unit (ICU) using a large Finnish database.

Methods: A national prospectively collected ICU data registry was used for analysis. All adult trauma admissions excluding isolated head trauma and burns registered from July 1999 to December 2006 were analyzed. Data from 22 ICUs were available. The non‐university‐affiliated units were categorized according to the number of beds and referral population as small, mid size and large. Acute physiology and chronic health evaluation (APACHE II)‐ and sequential organ failure assessment (SOFA)‐adjusted mortalities were compared between the units.

Results: There were 2067 trauma admissions that fulfilled the inclusion criteria; 38% were treated in the university hospitals, 26% in large non‐teaching ICUs, 20% in mid size ICUs and 15% in small ICUs. The crude hospital mortality was 5.6%, being 4.7% in university ICU and 6.6% in mid size ICU. In two subgroup analyses of severely ill trauma patients with APACHE II points >25 or SOFA score >8 points, respectively, hospital mortality was significantly lower in university ICUs.

Conclusions: University‐level hospitals were associated with better outcomes with critically ill trauma patients. These results can be used in planning future organization of trauma patient care in Finland.
Active cooling in traumatic brain‐injured patients: a questionable therapy?
Authors: GRÄNDE P.‐O. , REINSTRUP P. , ROMNER B.
Published: August 2009

Hypothermia is shown to be beneficial for the outcome after a transient global brain ischaemia through its neuroprotective effect. Whether this is also the case after focal ischaemia, such as following a severe traumatic brain injury (TBI), has been investigated in numerous studies, some of which have shown a tendency towards an improved outcome, whereas others have not been able to demonstrate any beneficial effect. A Cochrane report concluded that the majority of the trials that have already been published have been of low quality, with unclear allocation concealment. If only high‐quality trials are considered, TBI patients treated with active cooling were more likely to die, a conclusion supported by a recent high‐quality Canadian trial on children. Still, there is a belief that a modified protocol with a shorter time from the accident to the start of active cooling, longer cooling and rewarming time and better control of blood pressure and intracranial pressure would be beneficial for TBI patients. This belief has led to the instigation of new trials in adults and in children, including these types of protocol adjustments. The present review provides a short summary of our present knowledge of the use of active cooling in TBI patients, and presents some tentative explanations as to why active cooling has not been shown to be effective for outcome after TBI. We focus particularly on the compromised circulation of the penumbra zone, which may be further reduced by the stress caused by the difference in thermostat and body temperature and by the hypothermia‐induced more frequent use of vasoconstrictors, and by the increased risk of contusional bleedings under hypothermia. We suggest that high fever should be reduced pharmacologically.
Effect of P6 acustimulation on post‐operative nausea and vomiting in patients undergoing a laparoscopic cholecystectomy
Authors: FREY U. H. , FUNK M. , LÖHLEIN C. , PETERS J.
Published: August 2009

Background
Non‐pharmacologic techniques such as electrical acustimulation may mitigate post‐operative nausea and vomiting (PONV). The primary purpose of this study was to investigate the effectiveness of acustimulation on attenuating PONV. Moreover, we tested whether a pre‐ or a post‐induction application of acustimulation results in differences in PONV reduction.

Methods
In this prospective, double‐blind, randomized, controlled trial, we studied 200 patients undergoing a laparoscopic cholecystectomy during propofol (induction) fentanyl/isoflurane/atracurium (maintenance) anaesthesia. In the acustimulation group (n=101), subdivided into groups with pre‐induction (n=57) and post‐induction (n=44) acustimulation, an active ReliefBand® device was placed at the P6 acupoint. In the sham group (n=99), also subdivided into pre‐induction (n=55) or post‐induction (n=44) groups, an inactive device was applied instead. The ReliefBand® remained in place for 24 h after surgery. Nausea and vomiting/retching were recorded at 2, 6, and 24 h post‐operatively.

Results
The incidence of early nausea (up to 2 h) was significantly lower in the acustimulation than in the sham group (29% vs. 42%; P=0.043). No significant effect could be detected for retching/vomiting. Moreover, acustimulation showed no effect on PONV after 6 and 24 h. Risk factor analysis (female gender, non‐smoker, history of PONV/motion sickness, and post‐operative morphine usage) revealed a relative reduction in risk of 40% for nausea (P=0.021) and 55% for retching/vomiting (P=0.048) in patients with three or four risk factors present. The timing of (pre‐ vs. post‐induction) acustimulation had no significant effect on PONV reduction.

Conclusion
Acustimulation at the P6 acupoint reduces early nausea, but not vomiting, after laparoscopic cholecystectomy, irrespective of its pre‐ or post‐induction application.
Life‐threatening systemic toxicity and airway compromise from a common European adder bite to the tongue
Authors: HOEGBERG L. C. G. , JESSEN C. L. , LAMBERTSEN K. , STENSBALLE J.
Published: August 2009

A 24‐year‐old man was bit on the tongue by a European common adder. Within 15 min following envenomation, he experienced tongue swelling, hypotension and impaired consciousness. Antihistamine, corticosteroid and crystalloids were administered. Within 105 min of envenomation, increasing oral, pharyngeal and facial oedema compromised the airway, leading to respiratory failure, concomitant with circulatory failure related to hypoxaemia and systemic toxic effects. Acute tracheotomy secured the airway, and two doses of antivenom successfully treated the systemic, toxic effects. The reaction was severe due to rapid and suspected high‐dose uptake of venom, underlining the need for early advanced symptomatic treatment with airway control and early and eventually repeated dosing of antivenom.
The GlideScope Ranger® video laryngoscope can be useful in airway management of entrapped patients
Authors: NAKSTAD A. R. , SANDBERG M.
Published: August 2009

Background: Airway management of entrapped patients is challenging and alternatives to endotracheal intubation with a Macintosh laryngoscope must be considered. In this study, the GlideScope Ranger® video laryngoscope has been evaluated as an alternative to standard laryngoscopy.

Methods: Eight anaesthesiologists from a Helicopter Emergency Medical Service intubated the trachea of a Laerdal SimMan® manikin using the studied laryngoscopes in two scenarios: (A) unrestricted access to the manikin in an ambulance and (B) no access from the head end, simulating an entrapped patient. The time used to secure the airway and the scored level of difficulty were the main variables.

Results: In scenario A, all anaesthesiologists managed to secure the airway using both techniques within the 60‐s time limit. In scenario B, all secured the airway when using the video laryngoscope, while 50% succeeded with endotracheal intubation using the Macintosh laryngoscope. The difference in the success rate was statististically significant (P=0.025). There were no significant differences in the time spent on endotracheal intubation in the two scenarios or between the devices. All stated that the availability of a video laryngsoscope would make drug‐facilitated intubation a realistic alternative when access to patients is limited. The lack of visual control when using the Macintosh laryngoscope excludes this technique in real‐life settings.

Conclusion: This study suggests that the Glidescope Ranger® may be merited in situations requiring endotracheal intubation by an experienced intubator in patient entrapment. Further studies are required to clarify whether performance in patients mimics that in a manikin.
Role of leaf glandular trichomes of melon plants in deterrence of Aphis gossypii Glover
Authors: Sarria Emilio , Palomares‐Rius Francisco J. , López‐Sesé Ana I. , Heredia Antonio , Gómez‐Guillamón Maria L.
Published: August 2009

External characteristics of the leaf epidermis and their effects on behaviour of Aphis gossypii Glover were evaluated in two Cucumis melo L. genotypes, ‘Bola de Oro’ (aphid susceptible) and TGR‐1551 (aphid resistant) in order to explore their role in the early rejection of TGR‐1551 by this aphid. No differential effects of epicuticular waxes on aphid behaviour were observed. The type, distribution and number of trichomes on melon leaves were also studied. Pubescence in melon, measured as the number of non‐glandular trichomes per cm2, was not sufficient to prevent aphid settling. However, there was a high density of type I glandular trichomes on leaves of the aphid‐resistant genotype. According to microscopic observations and stain testing, these trichomes store and secrete phenols and flavonoids. Free‐choice tests were conducted to determine the effect of these glandular trichomes on A. gossypii preference, revealing that aphids reject leaf disks of TGR‐1551 from the onset of the experiment. Additional experiments after removal of leaf type I glandular trichome exudates showed that A. gossypii preferred washed TGR‐1551 leaf disks over unwashed disks, while this effect was not observed in experiments using washed and unwashed ‘Bola de Oro’ leaf disks. These results suggest that a high density of glandular trichomes and chemicals secreted by them deter A. gossypii and disturb aphid settling on TGR‐1551.