bluuebunny/crossref_metadata_2025 huggingface dataset:
Interrater reliability between scorers from eight European sleep laboratories in subjects with different sleep disorders
Authors: Danker‐Hopfe Heidi , Kunz D. , Gruber G. , Klösch G. , Lorenzo J. L. , Himanen S. L. , Kemp B. , Penzel T. , Röschke J. , Dorn H. , Schlögl A. , Trenker E. , Dorffner G.
Published: October 2010

Interrater variability of sleep stage scorings is a well‐known phenomenon. The SIESTA project offered the opportunity to analyse interrater reliability (IRR) between experienced scorers from eight European sleep laboratories within a large sample of patients with different (sleep) disorders: depression, general anxiety disorder with and without non‐organic insomnia, Parkinson's disease, period limb movements in sleep and sleep apnoea. The results were based on 196 recordings from 98 patients (73 males: 52.3 ± 12.1 years and 25 females: 49.5 ± 11.9 years) for which two independent expert scorings from two different laboratories were available. Cohen's κ was used to evaluate the IRR on the basis of epochs and intraclass correlation was used to analyse the agreement on quantitative sleep parameters. The overall level of agreement when five different stages were distinguished was κ = 0.6816 (76.8%), which in terms of κ reflects a ‘substantial’ agreement (Landis and Koch, 1977). For different groups of patients κ values varied from 0.6138 (Parkinson's disease) to 0.8176 (generalized anxiety disorder). With regard to (sleep) stages, the IRR was highest for rapid eye movement (REM), followed by Wake, slow‐wave sleep (SWS), non‐rapid eye movement 2 (NREM2) and NREM1. The results of regression analysis showed that age and sex only had a statistically significant effect on κ when the (sleep) stages are considered separately. For NREM2 and SWS a statistically significant decrease of IRR with age has been observed and the IRR for SWS was lower for males than for females. These variations of IRR most probably reflect changes of the sleep electroencephalography (EEG) with age and gender.
Rapid eye movement activity before spontaneous awakening in elderly subjects
Authors: Ficca G. , Scavelli S. , Fagioli I. , Gori S. , Murri L. , Salzarulo P.
Published: October 2010

Previous research in young subjects found that rapid eye movement (REM) density is higher in those REM phases which are followed by an awakening (REM‐W) than in those preceding NREM (REM‐N), suggesting a ‘gating role’ of REM sleep toward the awakening. It is not yet known whether this evidence is maintained in elderly subjects, who display, relative to young subjects, more awakenings, different sleep states from which the awakenings come (NREM in a high proportion of cases) and a general impairment of rapid eye movement activity (REMA). To investigate this issue, we have compared in three different age groups (young, old and ‘old old’ subjects) the features of REMA, including REM density and the amount and duration of REM bursts, between REM‐W and REM‐N. Whereas in the young REM density is higher in REM‐W than in REM‐N, this difference is already reduced in the old group and fully cancelled in the old old subjects. The evidence that old individuals spontaneously wake up despite the absence of an increase of REMA could imply that in the aged awakening is not preceded by an increase of the arousal level (expressed in REM sleep by the REMA). The similar duration of REM bursts for REM‐W and REM‐N in both groups of old subjects suggests that with age a marked impairment occurs in the organizational aspects of REMs, independently from the following state.
Corticospinal excitability and sleep: a motor threshold assessment by transcranial magnetic stimulation after awakenings from REM and NREM sleep
Authors: Bertini Mario , Ferrara Michele , De Gennaro Luigi , Curcio Giuseppe , Fratello Fabiana , Romei Vincenzo , Pauri Flavia , Maria Rossini Paolo
Published: October 2010

Transcranial magnetic stimulation (TMS) is a recently established technique in the neurosciences that allows the non‐invasive assessment, among other parameters, of the excitability of motor cortex. Up to now, its application to sleep research has been very scarce and because of technical problems it provided contrasting results. In fact delivering one single suprathreshold magnetic stimulus easily awakes subjects, or lightens their sleep. For this reason, in the present study we assessed motor thresholds (MTs) upon rapid eye movement (REM) and non‐rapid eye movement (NREM) sleep awakenings, both in the first and in the last part of the night. Taking into account that a full re‐establishment of wake regional brain activity patterns upon awakening from sleep needs up to 20–30 min, it is possible to make inferences about the neurophysiological characteristics of the different sleep stages by analyzing the variables of interest immediately after provoked awakenings. Ten female volunteers slept in the lab for four consecutive nights. During the first night the MTs were collected, following a standardized procedure: 5 min before lights off, upon stage 2 awakening (second NREM period), upon REM sleep awakening (second REM period), upon the final morning awakening (always from stage 2). Results showed that MTs increased linearly from presleep wakefulness to REM sleep awakenings, and from the latter to stage 2 awakenings. There was also a time‐of‐night effect on MTs upon awakening from stage 2, indicating that MTs decreased from the first to the second part of the night. The increase in corticospinal excitability across the night, which parallels the fulfillment of sleep need, is consistent with the linear decrease of auditory arousal thresholds during the night. The maximal reduction of corticospinal excitability during early NREM sleep can be related to the hyperpolarization of thalamocortical neurons, and is in line with the decreased metabolic activity of motor cortices during this sleep stage. On the contrary, the increase of MTs upon REM sleep awakenings should reflect peripheral factors. We conclude that our findings legitimate the introduction of the TMS technique as a new proper tool in sleep research.
Oleoyl‐CoA is not an immediate substrate for fatty acid elongation in developing seeds of Brassica napus
Authors: Hlousek‐Radojcic Alenka , Imai Hiroyuki , Jaworski Jan G.
Published: October 2010

The substrate specificity of fatty acid elongase was studied using an oil body fraction from developing seeds of Brassica napus. ATP was essential for high rates of elongase activity, but there was no apparent requirement for oleoyl‐CoA, oleic acid (18:1) or CoA. Furthermore, 14C from 18:1‐CoA was incorporated into eicosenoic (20:1) and erucic (22:1) acids at a much slower rate than 14C from malonyl‐CoA. Incubation of [14C]18:1‐CoA with the oil body fraction resulted in a rapid loss of [14C]18:1‐CoA into several lipid fractions whether in the absence or presence of ATP, but the loss of 18:1‐CoA had a comparatively small effect on the overall rate of elongation. Acyl‐CoAs were derivatized to their respective acylbutylamide and analyzed by gas chromatography‐mass spectrometry. This analysis of acyl‐CoAs demonstrated that there was no detectable 20:1‐CoA or 22:1‐CoA at 0 min incubation, while newly synthesized 20:1‐CoA and 22:1‐CoA were present at 10 min. Analysis of the %14C of the substrates and products of the elongation reaction revealed that the endogenous pool of 18:1‐CoA is quite small in elongase preparations. In addition, [14C]18:1‐CoA added to the incubation, although incorporated into lipids, was not significantly diluted by turnover or new synthesis. In contrast, the %14C of the 20:1‐CoA was two‐ to threefold less than that of the 18:1‐CoA. Taken together, these results indicate that the [14C]18:1 from the [14C]18:1‐CoA was diluted in an intermediate 18:1 pool and that the 18:1‐CoA was not the major donor of the acyl group to the elongase reaction.
A characterization of the MADS‐box gene family in maize
Authors: Mena Montaña , Mandel M. Alejandra , Lerner David R. , Yanofsky Martin F. , Schmidt Robert J.
Published: October 2010

Studies on distantly related dicot plant species have identified homeotic genes that specify floral meristem identity and determine the fate of floral organ primordia. Most of these genes belong to a family characterized by the presence of a structural motif, the MADS‐box, which encodes a protein domain with DNA‐binding properties. As part of an effort to understand how such genes may have been recruited during the evolution of flowers with different organ types such as those found in maize, two members of this gene family in maize, ZAG1 and ZAG2, have been characterized previously. Here, the isolation and characterization of four new members of this gene family, designated ZAP1, ZAG3, ZAG4 and ZAG5, are described and the genetic map position of these and 28 additional maize MADS‐box genes is determined. The first new member of this family appears to be the Zea mays ortholog of the floral homeotic gene APETALA1 (AP1) and has been designated ZAP1. One of these genes, ZAG4, is unusual in that its deduced protein sequence includes the MADS domain but lacks the K‐domain characteristically present in this family of genes. In addition, its copy number and expression varies among different inbreds. A large number of maize MADS‐box genes map to duplicated regions of the genome, including one pair characterized here, ZAG3 and ZAG5. These data underscore the complexity of this gene family in maize, and provide the basis for further studies into the regulation of floral organ morphogenesis among the grasses.
A novel lignin in poplar trees with a reduced caffeic acid/5‐hydroxyferulic acid O‐methyltransferase activity
Authors: Van Doorsselaere Jan , Baucher Marie , Chognot Emmanuelle , Chabbert Brigitte , Tollier Marie‐Thérèse , Petit‐Conil Michel , Leplé Jean‐Charles , Pilate Gilles , Cornu Daniel , Monties Bernard , Van Montagu Marc , Inzé Dirk , Boerjan Wout , Jouanin Lise
Published: October 2010

Lignin is a polymeric constituent of the cell wall that needs to be removed during the paper making process. Bi‐specific caffeic acid/5‐hydroxyferulic acid O‐methyltransferase (COMT) catalyses the O‐methylation of caffeic acid and 5‐hydroxyferulic acid to ferulic acid and sinapic acid, respectively. These compounds are intermediates in the biosynthesis of the lignin precursors. Therefore, COMTs are potential target enzymes for reducing the amount, or modifying the composition, of lignin in plants. Different antisense and sense constructs have been expressed of a gene encoding a COMT from poplar (Populus trichocarpa x P. deltoides) in a P. tremula x P. alba clone under the control of the cauliflower mosaic virus 35S promoter. From all analysed transformants, four lines transformed with an antisense construct had a reduced COMT activity. Two showed a 50% reduction of COMT activity, which altered only slightly the monomeric composition. In the two other transformants, the COMT activity was reduced by 95%. In the latter case, the syringyl/ guaiacyl ratio (S/G) was reduced by sixfold (due to a decrease of S and an increase of G), as analysed by thioacidolysis. A new component of lignin, the 5‐hydroxyguaiacyl residue, was detected among the thioacidolysis products. Moreover, in contrast to the white/yellow colour of wild‐type wood, the xylem of the transgenic lines with a 95% reduction of COMT activity was pale rose. A similar phenotype was observed in brown‐midrib mutants of maize and sorghum, known for their altered lignification. Although the lignin composition was consistently modified, the lignin content of the transgenic poplars was similar to that of the controls.
O‐glycosylation of a precursor to a sweet potato vacuolar protein, sporamin, expressed in tobacco cells
Authors: Matsuoka Ken , Watanabe Natsu , Nakamura Kenzo
Published: October 2010

Sporamin, a vacuolar protein of the sweet potato, is synthesized as a precursor that contains signal peptide and an N‐terminal propeptide that functions as a vacuolar targeting determinant. Sporamin, when expressed in tobacco cells, migrated as smeared bands on an SDS‐polyacrylamide gel. The smearing was due to O‐glycosylation of the precursor to sporamin. The smeared bands were stained by a glycan‐specific stain but no N‐glycosylation site was found in the amino acid sequence of the precursor to sporamin. The glycan attached to sporamin contained galactose and arabinose as major sugar components. Mutations that altered the Pro36 or Ser39 residue of the precursor to sporamin prevented glycosylation of the protein, and analysis by semiquantitative Edman degradation suggested that a glycan moiety was attached to Pro36 and, possibly, to Ser39. Pulse‐labeling and cell‐fractionation experiments revealed that the O‐glycosylation of the precursor to sporamin occurred in the Golgi apparatus. Thus, this modification serves as a good marker of the transport from the endoplasmic reticulum (ER) to the Golgi apparatus of the precursor to sporamin. Treatment of transformed tobacco cells with brefeldin A (BFA) caused the intracellular accumulation of prosporamin that did not migrate as smeared bands. Thus, it appeared that BFA inhibited the transport of the precursor to sporamin to the Golgi apparatus. This result provides the first biochemical evidence that BFA inhibits transport from the ER to the Golgi apparatus in plant cells.
Transposon tagging of the maize Glossy2 locus with the transposable element En/Spm
Authors: Tacke Eckhard , Korfhage Christian , Michel Detlef , Maddaloni Massimo , Motto Mario , Lanzini Simona , Salamini Francesco , Döring Hans‐Peter
Published: October 2010

The Glossy2 (Gl2) locus of maize is required for the formation of the epicuticular wax layer of young plants. gl2 mutant seedlings can be visually identified because of their glossy leaf surface which is different from the dull surface of wild‐type seedlings. The Gl2 locus was isolated by transposon tagging. Seven unstable mutations, gl2‐m2 to gl2‐m8, were induced in a parental strain carrying an active transposable Activator (Ac) element in the unstable wx‐m7 allele. Genetic tests on the gl2‐m2 allele indicated that it was not caused by the Ac element but by the insertion of the transposable element Enhancer/Suppressor‐Mutator (En/Spm). A SaIl restriction fragment segregating with the mutant phenotype was identified, by Southern analysis, using sequences from the En/Spm element as a probe. Part of the fragment was cloned and was shown to carry part of the unstable gl2‐m2 allele. These gl2 sequences were used to identify a genomic fragment carrying the wild‐type allele and to isolate its corresponding cDNA sequence. The predicted Glossy2 protein consists of 426 amino acids. No similar amino acid sequence was found in protein data banks and the biochemical function of the Gl2 gene product is still unknown. The wild‐type Gl2 transcript is found predominantly in juvenile leaves. The transcript level in the leaves of seedlings homozygous for a stable recessive gl2‐ref allele is hardly detectable.
The coat protein of potato virus X is a strain‐specific elicitor of Rx1‐mediated virus resistance in potato
Authors: Bendahmane Abdelhafid , Köhm Bärbel A. , Dedi Cynthia , Baulcombe David C.
Published: October 2010

The Rx1 gene in potato confers extreme resistance to potato virus X (PVX). To investigate the mechanism and elicitation of Rx resistance, protoplasts of potato cv. Cara (Rx1 genotype) and Maris Bard (rx1 genotype) were inoculated with PVX and tobacco mosaic virus (TMV). At 24 h post‐inoculation in Maris Bard protoplasts there was at least 100‐fold more PVX RNA than in protoplasts of Cara. TMV RNA accumulated to the same level in both types of protoplast. However, when the TMV was inoculated together with PVX the accumulation of TMV RNA was suppressed in the Cara (Rx1 genotype) protoplasts to the same extent as PVX. The Rx1 resistance also suppressed accumulation of a recombinant TMV in which the coat protein gene was replaced with the coat protein gene of PVX. It is therefore concluded that Rx1‐mediated resistance is elicited by the PVX coat protein, independently of any other proteins encoded by PVX. The domain of the coat protein with elicitor activity was localized by deletion and mutation analysis to the structural core of a non‐virion form of the coat protein.
Tobacco mitotic cyclins: cloning, characterization, gene expression and functional assay
Authors: Setiady Yulius Yulianto , Sekine Masami , Hariguchi Norimitsu , Yamamoto Takuo , Kouchi Hiroshi , Shinmyo Atsuhiko
Published: October 2010

Three cyclin cDNA clones (Ntcyc25, Ntcyc27, Ntcyc29) have been isolated from tobacco (Nicotiana tabacum) using the PCR cloning method. The encoded Ntcyc cyclins were highly homologous to mitotic cyclins. In synchronized tobacco suspension cultured cells, the mRNA levels of Ntcyc25 and Ntcyc27 were detectable through S, G2 and M phases, while the Ntcyc29 mRNA was detected from G2 to M phase. These expression patterns classified the Ntcyc25 and Ntcyc27 into A‐type cyclin and Ntcyc29 into B‐type cyclin. The three genes were expressed in growing tobacco cultured cells but ceased to be expressed when cells entered the stationary phase, indicating that the expression of these cyclin genes was well correlated with cell growth. The N‐terminal truncated Ntcyc25 and Ntcyc29 cDNAs were able to rescue G1 cyclin mutant of Saccharomyces cerevisiae, while the full‐length of Ntcyc27 protein could also partially perform G1 cyclin functions.
Multiple regulatory sites in the C‐terminal autoinhibitory domain of the plasma membrane H+‐ATPase
Authors: Olsson Anne , Johansson Fredrik , Sommarin Marianne , Larsson Christer
Published: October 2010

The H+‐ATPase activities of root and leaf plasma membranes from tobacco (Nicotiana tabacum) have been characterized with respect to Vmax, Km for ATP, pH dependence and activation involving the C‐terminal autoinhibitory domain. With root plasma membranes, addition of lysophosphatidylcholine (lyso‐PC) resulted in the expected increase in Vmax, a decrease in Km(ATP), and a shift in pH optimum to a more alkaline pH, typical for activation via the C‐terminal inhibitory domain. With leaf plasma membranes, however, Km(ATP) was relatively low and the pH optimum was around pH 7.0 before the addition of lyso‐PC and did not change upon addition of the activator, although Vmax increased twofold. Similar results were obtained with the in vivo activator fusicoccin. The results obtained with the leaf plasma membranes show that Vmax may be regulated independently of Km(ATP) and pH optimum, and suggest the presence of at least two regulatory sites within the C‐terminal autoinhibitory domain of the H+‐ATPase.
Isolation of two novel myb‐like genes from Arabidopsis and studies on the DNA‐binding properties of their products
Authors: Li Song Feng , Parish Roger W.
Published: October 2010

Two novel myb‐like genes (atmyb6 and atmyb7) were isolated from an Arabidopsis thaliana cDNA library. The entire proteins or the Myb domains encoded by the genes were expressed as fusion proteins in Escherichia coli. The DNA‐binding domain of the murine c‐Myb was also expressed in the same way for use in comparative studies. The fusion proteins were examined for their DNA‐binding activity using the animal c‐Myb DNA‐binding site (MBS) and the binding site of the maize P gene product (PBS). The Myb domain of Atmyb6 bound to PBS more efficiently than to MBS. Complete Atmyb6 and Atmyb7 proteins preferentially bound to PBS but not MBS. This suggests that the in vitro binding consensus sequences for both Atmyb6 and Atmyb7 are similar to PBS. The binding of the Myb domain of Atmyb6 to both PBS and MBS raises the possibility that the protein recognizes multiple sequences in vivo. The third α‐helix and three adjacent amino acids in the third repeat (R3) of c‐Myb were replaced with the analogous sequence of Atmyb6 to create a chimeric Myb protein. This chimeric protein bound to PBS with a low affinity but failed to bind to MBS. Thus the binding pattern of the chimeric Myb protein is similar to that of the Atmyb6. This result suggests that the last 20 amino acids in the R3 repeat of Atmyb6 play a major role in DNA‐binding.
Microdissection and amplification of coding sequences from a chromosome fragment restoring male fertility in alloplasmic male‐sterile tobacco.
Authors: Hernould Michel , Glimelius Kristina , Veuskens Jacky , Bergman Per , Mouras Armand
Published: October 2010

An alloplasmic male‐sterile line of tobacco, containing the nucleus of Nicotiana tabacum and the cytoplasm of Nicotiana repanda, is restored to fertility by introgression of an alien chromosome fragment obtained from the cytoplasm donor. To isolate the restorer gene(s), the alien chromosome fragment was microdissected from metaphase plates of the restored line. The microdissected chromosomes represented only 0.1 pg of DNA, which was amplified using a degenerate oligonucleotide‐primed PCR method (DOP‐PCR), from which a chromosome fragment specific library was created. Compared with previous strategies used for microcloning, a modified and improved method was developed by the subsequent isolation of expressed sequences. The library was screened with cDNA probes synthesized by reverse transcription and DOP‐PCR amplification (RT/DOP‐PCR), of total RNAs isolated from early developing restored and male‐sterile flower buds. By this strategy, transcribed DNA sequences specific for the restored line were cloned.
Overexpression of light‐dependent PORA or PORB in plants depleted of endogenous POR by far‐red light enhances seedling survival in white light and protects against photooxidative damage
Authors: Sperling Ulrich , Van Cleve Barbara , Frick Geneviève , Apel Klaus , Armstrong Gregory
Published: October 2010

The structurally related light‐dependent protochlorophyllide (Pchlide) oxidoreductases PORA and PORB mediate the only light‐requiring step in chlorophyll (Chl) biosynthesis in higher plants. Correlative evidence suggests that some in vivo functions of PORA and PORB may be unique, including a postulated photoprotective role for PORA. For example, wild‐type Arabidopsis thaliana seedlings grown in non‐photooxidative far‐red light (cFR) resemble those grown in white light (WL), but they are yellow and do not green normally thereafter in WL. This defect is accompanied by the absence of detectable PORA and reduced levels of PORB expression. Here, direct evidence is provided that the presence of POR, either as PORA or PORB, can confer photoprotection in plants. In contrast to the wild‐type, the plastids of transgenic PORA‐ or PORB‐overexpressing Arabidopsis seedlings grown in cFR possess extensive prolamellar bodies. Upon a subsequent shift to WL, POR‐overexpressing seedlings develop thylakoid membranes, accumulate large amounts of Chl and are viable at fluence rates lethal to the wild‐type. Intriguingly, the plastid membrane architectures of greening transgenic seedlings seem to depend on whether PORA or PORB has been overproduced. POR‐overexpressing seedlings shifted from cFR to WL of fluence rates from 20 to 500 μE m–2 sec–1 accumulate substantially higher amounts of Chl than does the wild‐type. Furthermore, the WL fluence rate that permits maximal Chl accumulation increases from 8 μE m–2 sec–1 in the wild‐type to 125 μE m–2 sec–1 in transgenic seedlings. POR overexpression during growth in cFR also correlates with a fourfold decrease in the steady‐state content of Pchlide, a potentially lethal photosensitizer.
A negative selection scheme based on the expression of cytosine deaminase in plastids
Authors: Serino Germán , Pal Maliga Pal
Published: October 2010

The enzyme cytosine deaminase (CD) encoded by codA catalyzes deamination of cytosine to uracil. CD is present in prokaryotes and in many eukaryotic micro‐organisms, but is absent in higher plants. 5‐fluorocytosine (5FC) is metabolized in CD‐expressing cells, causing cellular death. A chimeric codA has been introduced into the tobacco plastid genome and 5FC was used to select against tissue culture cells and seedlings expressing CD. This negative selection scheme will be useful in identifying nuclear genes which control plastid gene expression in higher plants.
The plasma membrane aquaporin NtAQP1 is a key component of the leaf unfolding mechanism in tobacco
Authors: Siefritz Franka , Otto Beate , Bienert Gerd Patrick , Van Der Krol Alexander , Kaldenhoff Ralf
Published: October 2010

Epinastic leaf movement of tobacco is based on differential growth of the upper and lower leaf surface and is distinct from the motor organ‐driven mechanism of nyctinastic leaf movement of, for example, mimosa species. The epinastic leaf movement of tobacco is observed not only under diurnal light regimes but also in continuous light, indicating a control by light and the circadian clock. As the transport of water across membranes by aquaporins is an important component of rapid plant cell elongation, the role of the tobacco aquaporin Nt aquaporin (AQP)1 in the epinastic response was studied in detail. In planta NtAQP1‐luciferase (LUC) activity studies, Northern and Western blot analyses demonstrated a diurnal and circadian oscillation in the expression of this plasma membrane intrinsic protein (PIP)1‐type aquaporin in leaf petioles, exhibiting peaks of expression coinciding with leaf unfolding. Cellular water permeability of protoplasts isolated from leaf petioles was found to be high in the morning, i.e. during the unfolding reaction, and low in the evening. Moreover, diurnal epinastic leaf movement was shown to be reduced in transgenic tobacco lines with an impaired expression of NtAQP1. It is concluded that the cyclic expression of PIP1‐aquaporin represents an important component of the leaf movement mechanism.
Establishing an efficient Ac/Ds tagging system in rice: large‐scale analysis of Ds flanking sequences
Authors: Kolesnik Tatiana , Szeverenyi Ildiko , Bachmann Doris , Kumar Chellian Santhosh , Jiang Shuye , Ramamoorthy Rengasamy , Cai Minnie , Ma Zhi Gang , Sundaresan Venkatesan , Ramachandran Srinivasan
Published: October 2010

A two‐element Activator/Dissociation (Ac/Ds) gene trap system was successfully established in rice (Oryza sativa ssp. japonica cv. Nipponbare) to generate a collection of stable, unlinked and single‐copy Ds transposants. The germinal transposition frequency of Ds was estimated as an average of 51% by analyzing 4413 families. Study of Ds transposition pattern in siblings revealed that 79% had at least two different insertions, suggesting late transposition during rice development. Analysis of 2057 Ds flanking sequences showed that 88% of them were unique, whereas the rest within T‐DNA. The insertions were distributed randomly throughout the genome; however, there was a bias toward chromosomes 4 and 7, which had two times as many insertions as that expected. A hot spot for Ds insertions was identified on chromosome 7 within a 40‐kbp region. One‐third of Ds flanking sequences was homologous to either proteins or rice expressed sequence tags (ESTs), confirming a preference for Ds transposition into coding regions. Analysis of 200 Ds lines on chromosome 1 revealed that 72% insertions were found in genic region. Anchoring of more than 800 insertions to yeast artificial chromosome (YAC)‐based EST map showed that Ds transposes preferentially into regions rich in expressed sequences. High germinal transposition frequency and independent transpositions among siblings show that the efficiency of this system is suitable for large‐scale transposon mutagenesis in rice.
GIP, a Petunia hybrida GA‐induced cysteine‐rich protein: a possible role in shoot elongation and transition to flowering
Authors: Ben‐Nissan Gili , Lee Jung‐Youn , Borohov Amihud , Weiss David
Published: October 2010

The Petunia hybrida GA‐induced proteins (GIPs) belong to a large group of proteins identified in numerous plant species. These proteins share a similar C‐terminal region containing 12 cysteine residues in conserved positions. To date, the function of these proteins remains unclear. We previously found that GIP1 expression coincides with cell elongation in stems and flowers and is induced by gibberellic acid (GA3). Transient expression of a GIP1:green fluorescent protein (GFP) fusion in tobacco bright yellow 2 (BY2) cells and immunoblot analyses suggest microsomal compartmentalization with possible endoplasmic reticulum (ER) localization. However, the polyclonal anti‐GIP1 antibodies also reacted with proteins extracted from the cell wall. Three novel GIP homologs, GIP2, GIP4, and GIP5, were isolated. While GIP4, similar to GIP1, is putatively localized to the ER membrane, the cleavable hydrophobic N‐terminal sequences of GIP2 and GIP5 suggest cell wall localization. GIP1 and GIP2 are expressed during cell elongation, whereas GIP4 and GIP5 are expressed during cell division; nevertheless, they all were induced by GA3. We generated transgenic petunia in which we repressed the putative cell wall protein GIP2. The transgenic plants exhibited late flowering and reduced stem elongation. These phenotypic alterations were found under low, but not moderate‐high temperatures, suggesting functional redundancy under normal growth conditions. The expression pattern and cellular localization of GIP2, its regulation by GA, and the phenotype of the transgenic plants suggest a role in GA‐mediated cell elongation and transition to flowering.
Cytoplasmic regulation of the accumulation of nuclear‐encoded proteins in the mitochondrial proteome of maize
Authors: Hochholdinger Frank , Guo Ling , Schnable Patrick S.
Published: October 2010

Mitochondria from normal (NA)‐ and Texas (T)‐cytoplasm maize (Zea mays L.) were purified from unpollinated ears via Percoll centrifugation. Approximately 300 mitochondrial proteins were resolved using two‐dimensional (2‐D) electrophoresis. The 197 most abundant proteins were analyzed by matrix‐assisted laser desorption lonization time‐of‐flight (MALDI‐ToF) mass spectrometry involving overlapping pH gradients (pH 4–7 and 6–9). Database searches identified 58 genes that encode 100 of these protein spots. Functions could be predicted for 38 of the 58 genes (66%). All but one of these genes are located in the nuclear genome. Thirteen per cent of the analyzed protein spots (25 out of 197) exhibited at least a threefold difference in accumulation between the mitochondrial proteomes of NA‐ or T‐cytoplasm maize plants that had essentially identical nuclear genomes. As most of these proteins were nuclear‐encoded, these findings demonstrate that the genotype of a mitochondrion can regulate the accumulation of the nuclear‐encoded fraction of its proteome. About half (27 out of 58) of the maize mitochondrial proteins identified in this study were not recovered in previous analyses of the Arabidopsis and rice mitochondrial proteomes.
New plant growth‐modifying properties of the Agrobacterium T‐6b oncogene revealed by the use of a dexamethasone‐inducible promoter
Authors: Grémillon Louis , Helfer Anne , Clément Bernadette , Otten Léon
Published: October 2010

Agrobacterium 6b oncogenes induce tumours on Nicotiana glauca and enations and associated modifications in transgenic N. tabacum plants. 2x35S‐AB‐6b tobacco rootstocks produced a graft‐transmissible factor that induced enations in wild‐type scions; the nature of this enation factor remains to be identified. Here, we report on the properties of tobacco plants carrying a dexamethasone‐inducible T‐6b gene (dex‐T‐6b). Induction with dex led to complex growth modifications, many of which have not been reported previously. Modifications were only found in growing tissues; mature tissues remained unaffected. Growth could be either stimulated or inhibited. Dex induction of young plants led to morphogenetic gradients that included enations, tubular leaves and fragmented leaf primordia. Root elongation was increased or slowed down, while radial root growth was strongly enhanced. Additional cell divisions were found in the root pericycle and vasculature. Enation factor import from mature tissues did not have the same effects on growing tissues as local T‐6b synthesis: normal scions grafted on induced dex‐T‐6b rootstocks formed enations, whereas local dex‐T‐6b induction at the shoot apex led to numerous dark‐green spots on the abaxial side of the leaves. In leaf patch assays, the 23‐kDa T‐6b protein was found to move through leaves and to enter the vascular system. This and the fact that rootstocks of spontaneous tobacco enation mutants did not modify wild‐type scions contrary to 6b plants indicate that the 6b protein might be the enation factor.
Rac regulation of NtrbohD, the oxidase responsible for the oxidative burst in elicited tobacco cell
Authors: Morel Johanne , Fromentin Jérome , Blein Jean‐Pierre , Simon‐Plas Françoise , Elmayan Taline
Published: October 2010

Five cDNAs encoding Rac protein homologues to the Rho‐related proteins from plants (Rop) were isolated in tobacco, and the function of one of them, Ntrac5, was studied. The Ntrac5 mRNA is repressed when tobacco leaves and cells are treated with the fungal elicitor cryptogein. Tobacco cells were transformed with sense constructs of Ntrac5 or Ntrac5V15, encoding the native GTP/GDP‐bound form of this Rac protein homologue or the constitutively active mutant in its GTP‐bound form, respectively. Immunological studies indicate that the corresponding protein is continuously located on the plasma membrane (PM). Both types of transformed cells show the same extra‐cellular alkalinization as the control, but a high decrease in the active oxygen species (AOS) production after elicitation with cryptogein. Moreover, the regulation of NtrbohD, the oxidase involved in AOS production upon elicitation, is affected at both transcriptional and translational levels in cells overexpressing Ntrac5. Thus, Ntrac5 could be considered as a negative regulator of NtrbohD.
Feeding problems in infants with gastro‐oesophageal reflux disease: A controlled study
Authors: Mathisen B , Worrall L , Masel J , Wall C , Shepherd RW
Published: October 2010

Objective:Gastro‐oesophageal reflux disease (GORD) in infants is commonly associated with feeding problems but has not been subject to systematic controlled study. We evaluated feeding, dietary, behavioural data obtained from systematic objective studies of six‐month old infants with and without GORD.

Methods:Infants with GORD (defined by 24‐h pH monitoring,n= 20), and age, gender, gestation, and socio‐economic matched healthy infants (n= 20) had standardised assessments of dietary intake, oromotor function by videoanalysis (Feeding Assessment Schedule, FAS), and infant feeding behaviour by Testers and Maternal Ratings (TRIB and MRIB). Videofluoroscopic analyses of swallowing was undertaken in 11/20 GORD infants and analysed by standardised paediatric check list.

Results:Compared with control data: GORD infants had significantly lower energy intakes; the FAS showed GORD infants to have significantly fewer adaptive skills and readiness behaviour for solids, significantly more food refusal and food loss; the TRIB showed GORD infants to be significantly more demanding and difficult with feeds; and the MRIB revealed that mothers of GORD infants had significantly more negative feelings, significantly less enjoyment of feeds, and reported significantly more crying behaviour. On videofluoroscopy, oral‐preparatory and oral phase problems predominated, particularly with solids, silent aspiration occurred during the pharyngeal phase in 2/11, and delayed oesophageal transit occurred in 4/11.

Conclusions:Feeding problems affecting behaviour, swallowing, food intake, and mother–child interaction occur in infants with GORD, who displayed a lack of development of age‐appropriate feeding skills. The contribution of feeding problems to morbidity in GORD in infants has been underestimated in the past.
Six‐month follow‐up of children with obstructive sleep apnoea
Authors: Harvey JMM , O’Callaghan MJ , Wales PD , Harris MA , Masters IB
Published: October 2010

Objectives: This study examined prospectively changes in development, temperament and sleep related behaviour in children referred for obstructive sleep apnoea (OSA) and polysomnographic sleep study, some of whom had surgical intervention.

Methodology: Using a prospective cohort study design, parents of 56 children referred for OSA completed sleep and temperament questionnaires and their child was assessed developmentally at the time of the polysomnographic sleep study. Forty (72%) of the children were neurologically normal. At 6 months, 42 children were reassessed using sleep and temperament questionnaires and a developmental assessment. After excluding the primary snorers, subjects were categorised as having had intervention (n = 24) or not (n = 15), and differences over the 6‐month period in Griffiths scores, temperament and sleep related behaviour were examined.

Results: Regardless of intervention status, there was an improvement in night‐time and day‐time sleep behaviour for the total group, though the extent of improvement was more marked in the intervention group. For the neurologically normal children, improvement in the sleep behaviour was only significant for the intervention group (P < 0.05). Intervention did not result in any significant changes in Griffiths developmental score or temperament.

Conclusion: Surgical intervention improves sleep behaviour in children though not temperament or development.
Cisapride and Caesarean section: Their role in babies with gastroschisis
Authors: Kishore Kumar R , Shi ECP , Duffy B
Published: October 2010

Objective: The objective of this study was to compare the neonatal postoperative course and morbidity for patients with gastroschisis who received cisapride with those who did not receive cisapride.

Study design: Data were obtained by review of the medical records of all the patients with gastroschisis who were admitted to Sydney Children’s Hospital between January 1984 and December 1995. Data were compared between 15 babies who received cisapride with 27 who did not. The mode of delivery and outcome of babies in whom gastroschisis was diagnosed antenatally was compared with those who were diagnosed at birth.

Results: Duration to the commencement of feeds, attainment of full feeds and the length of hospital stay were not statistically different between these two groups, with or without cisapride (p = ≥ 0.1). There were more elective Caesarean sections in the antenatally diagnosed group compared to those detected at birth and the outcome of these two groups showed no statistically significant difference.

Conclusions: Our study identified no benefit from cisapride therapy in babies with gastroschisis and also there was no benefit from elective Caesarean section for babies with antenatal diagnosis of gastroschisis.
Randomised controlled trials addressing Australian Aboriginal health needs: A systematic review of the literature
Authors: Morris PS
Published: October 2010

Objective: To describe the frequency and design of controlled clinical trials specifically addressing the health needs of Aboriginal Australians.

Methodology: Electronic searching of Medline, the Australasian Medical Index, the Aboriginal and Torres Strait Islander health bibliographic database, and handsearching of Aboriginal Health: an annotated bibliography. Studies that met the following selection criteria were included: i) addressed an Aboriginal health problem, ii) had a formal description of methods and results, and iii) compared the health effects of an intervention with a concurrent control group. All summary data were extracted by a single author.

Results: Only 13 studies were identified. Nine were randomised controlled trials and four were controlled trials but not randomised. Only one of these involved adults, which is unexpected. Although important Aboriginal child health issues were addressed in 12 of the 13 studies, most were undertaken many years ago and may not be familiar to Australian paediatricians. The majority appeared to be designed appropriately. Interestingly, the two studies not published in the medical literature had the largest sample size. There is no evidence that the number of published clinical trials involving Aboriginal Australians is increasing, or that long‐term applied clinical research programmes have been established.

Conclusion: There is a profound lack of well‐designed studies assessing medical interventions. This is further evidence that Australia has failed to develop a research infrastructure able to inform health care in Aboriginal communities. Adults appear especially disadvantaged.
Aetiological factors and development in subjects with obstructive sleep apnoea
Authors: Harvey JMM , O’Callaghan MJ , Wales PD , Harris MA , Masters IB
Published: October 2010

Objective: To examine whether maternal pregnancy complications, adverse birth events, respiratory illnesses, or developmental difficulty were increased in neurologically normal children with obstructive sleep apnoea (OSA) and whether severity of OSA adversely affects the child’s development and temperament.

Methodology: Maternal report of perinatal events, respiratory illness and developmental difficulty in 37 children with OSA was contrasted with a comparison group (n = 67). Children with OSA were assessed developmentally (Griffiths Scales), had a parental rating of temperament (Australian Temperament Scale) and attended an overnight polysomnographic sleep study.

Results: Children with OSA had an increased prevalence of adverse maternal pregnancy and perinatal events, respiratory disease and developmental concerns. Limited associations were found between the severity of OSA and development or temperament difficulty.

Conclusions: This study suggests a relationship between OSA, though not its severity, and pre/perinatal adversity and child development. Polysomnographic and detailed developmental assessment of community‐based samples of children with OSA and control children are necessary to confirm these findings.
Alcohol use by parents who present their child to a paediatric emergency department
Authors: Sharma S , Llewelyn K , Jureidini J
Published: October 2010

Objectives: Parental alcohol misuse exposes children to risk, but is poorly identified in paediatric settings. We aimed to (i) measure the alcohol use in parents of a sample of children attending emergency and (ii) assess the quality of documentation of this alcohol use by paediatric residents.

Method: Parents of children presenting to the Emergency Department of a children’s hospital were interviewed regarding their alcohol use, and each completed an AUDIT CORE questionnaire. The child’s case notes were reviewed for documentation of parental alcohol use.

Results: One hundred and ninety‐three parents were interviewed. Although per capita alcohol use for both sexes, and hazardous drinking for females were found to be significantly less than general population figures, rates of hazardous drinking were high for fathers. Yet only 1% of case notes reviewed contained any documentation of parental alcohol use.

Conclusion: This small study suggests that although average alcohol intake amongst parents presenting their children to a paediatric emergency department may be lower than for the general population, there is a significant prevalence of undetected hazardous drinking amongst parents.
Atrial septal defect in Malta
Authors: Grech V
Published: October 2010

Objective: To study diagnostic and surgical trends in atrial septal defect (ASD) in a population‐based study, and estimate birth prevalence and spontaneous closure rates.

Methodology: All patients in Malta diagnosed as having ASD and born between 1990 and 1994 were identified from various sources. This took place in the setting of a regional hospital supplying diagnostic services for the entire population. Echocardiographic follow‐up was also undertaken for lesions not requiring intervention.

Results: A total of 190 patients born in this period were diagnosed as having ASD. Age at diagnosis and age at surgery have decreased significantly over the period under study (P < 0.0001). The mode of diagnosis has become entirely noninvasive, and the perioperative mortality decreased dramatically over time. For the period 1990–94, the incidence at birth for defects not requiring intervention was 2.0/1000 live births, defects requiring intervention 0.4/1000 live births. A total of 92% of 50 defects not requiring intervention closed spontaneously, and the remainder had spontaneously decreased in size on follow‐up.

Conclusions: ASD is a relatively benign malformation in which early and noninvasive diagnosis can be achieved, with an extremely low interventional mortality.
Confirmation that Child Behavior Checklist clinical scales discriminate juvenile mania from attention deficit hyperactivity disorder
Authors: Hazell PL , Lewin TJ , Carr VJ
Published: October 2010

Objective: To determine whether boys meeting diagnostic criteria for juvenile mania and attention deficit hyperactivity disorder (mania‐ADHD) may be distinguished from boys with ADHD alone on a range of clinical and family variables.

Methodology: Boys aged 9–13 years with mania‐ADHD (n = 25), ADHD alone (n = 99), or no psychiatric diagnosis (n = 27) were compared on parent and teacher report Child Behavior Checklists (CBCL) and Conners Questionnaires, self‐report CBCLs, patterns of comorbidity, intellectual functioning, and family variables.

Results: Mania‐ADHD subjects had significantly higher mean ratings than ADHD only subjects on the parent CBCL for the Withdrawn, Thought Problems, Delinquent Behavior and Aggressive Behavior scales and significantly higher rates of comorbid depression, anxiety and psychotic symptoms. Other variables did not distinguish the mania‐ADHD and ADHD only groups.

Conclusions: These data confirm previous research indicating that the CBCL may be used to assist in the clinical identification of manic children.
Seroprevalence of Helicobacter pylori infection among children and their parents in eastern Turkey
Authors: Yilmaz E , Doğan Y , Gürgöze MK , Ünal S
Published: October 2010

Objectives: To determine the seroprevalence of Helicobacter pylori infection among healthy children in eastern Turkey, to assess risk factors for seroconversion, and to assess the role of parental infection status in the transmission of H. pylori.

Methods: A commercial enzyme immunoassay test for IgG to H. pylori was used. Demographic information obtained included age, gender, socio‐economic status and living conditions.

Results: One hundred and fifty‐two (43.9%) of 346 children were seropositive. There was no significant difference in rates of seroprevalence between boys and girls (P > 0.05), but a significant rise was noted with increasing age (P < 0.001). We found a strong inverse correlation between family income and seropositivity (P < 0.001). Seroprevalence also varied significantly with the educational level of the mother (P < 0.001). There were no significant differences in seroprevalence associated with place of residence, water supply and the number of persons per room (P > 0.05). The prevalence of infection in the corresponding parents was 85.4% for mothers and 76.3% for fathers. Seroprevalence was higher in children whose mother was infected (P < 0.001).

Conclusions: In eastern Turkey, as in other developing countries, H. pylori infection occurs early and increases with age. Infected parents, especially infected mothers, may have a important role in the transmission of H. pylori within families.