bluuebunny/crossref_metadata_2025 huggingface dataset:
Thyroid function and autoantibody status among women who spontaneously deliver under 35 weeks of gestation
Authors: Tierney Kelly , Delpachitra Pavitra , Grossmann Mathis , Onwude Joseph , Sikaris Ken , Wallace Euan M. , Hamilton Emma J. , Tong Stephen
Published: May 2009

Objective  Recent studies report an association between maternal thyroid auto‐antibodies and preterm birth. None have made the important distinction between spontaneous and iatrogenic preterm birth. We investigated the association between spontaneous preterm birth before 35‐week gestation and both thyroid function and auto‐antibody status.

Design  Retrospective nested case–control study, samples retrieved from a biobank of around 8000 samples.

Patients  Forty‐six cases of spontaneous preterm birth <35‐weeks gestation compared with 124 cases who delivered at 40 weeks gestation, matched for maternal age and duration of serum storage. All resulted in a singleton liveborn.

Measurements  First trimester serum levels of TSH, free T4, free T3, anti‐thyroid peroxidase (anti‐TPO) and anti‐thyroglobulin (anti‐Tg) antibodies.

Results  Women who had a preterm birth delivered at a median gestational age of 33 (+1 day) weeks gestation (controls 40 + 3 weeks) and had significantly lower birthweight infants [mean (range) weight 1965 (581–3117) grams compared with controls 3526 (2685–4760) grams]. Median levels of anti‐thyroglobulin and anti‐TPO antibodies were similar in the two groups (Anti‐Tg: 3 IU/ml (Range 1–316) v 3 IU/ml (1–691); Anti‐TPO 8 IU/ml (3–73) v 9 IU/ml (3–499); P ≥ 0·18 for both comparisons, compared using three statistical models). In the preterm group, the median TSH level was 0·70 mIU/l (range 0·04–3·84), similar to controls (0·88 (0·01–2·87), P = 0·1). Free T3 and T4 levels were also similar in both groups.

Conclusions  Thyroid function and thyroid autoantibody status among women who have a spontaneous preterm delivery before 35 weeks gestation are not different from women delivering at term.
Thermal acclimation of swimming performance in newt larvae: the influence of diel temperature fluctuations during embryogenesis
Authors: Měráková Eva , Gvoždík Lumír
Published: May 2009

1. Thermal acclimation is one of the basic strategies by which organisms cope with thermal heterogeneity of the environment. Under predictable variation in environmental temperatures, theory predicts that selection favours acclimation of thermal performance curves over fixed phenotypes.

2. We examined the influence of diel fluctuations in developmental temperatures on the thermal sensitivity of the maximal swimming capacity in larvae of the alpine newt,Triturus alpestris.

3. We incubated newt eggs under three thermal regimes with varying daily amplitudes (1, 5 and 9 °C) and similar means (17·6–17·9 °C), and accordingly we measured the swimming speed of hatched larvae at three experimental temperatures (12, 17 and 22 °C), which they would normally experience in their natural habitat.

4. Embryonic development under low and middle temperature fluctuations produced larvae with similar swimming speeds across experimental temperatures. In contrast, the most fluctuating regime induced development of phenotypes, which at 12 °C swam faster than larvae developed under moderate diel fluctuations.

5. Our results provide evidence that diel temperature fluctuations induce acclimation of thermal dependence of locomotor performance. In ectotherms experiencing diel cycles in environmental temperatures, this plastic response may act as an important pacemaker in the evolution of thermal sensitivity.
Linking forest fires to lake metabolism and carbon dioxide emissions in the boreal region of Northern Québec
Authors: MARCHAND DELPHINE , PRAIRIE YVES T. , Del GIORGIO PAUL A.
Published: May 2009

Natural fires annually decimate up to 1% of the forested area in the boreal region of Québec, and represent a major structuring force in the region, creating a mosaic of watersheds characterized by large variations in vegetation structure and composition. Here, we investigate the possible connections between this fire‐induced watershed heterogeneity and lake metabolism and CO2 dynamics. Plankton respiration, and water–air CO2 fluxes were measured in the epilimnia of 50 lakes, selected to lie within distinct watershed types in terms of postfire terrestrial succession in the boreal region of Northern Québec. Plankton respiration varied widely among lakes (from 21 to 211 μg C L−1 day−1), was negatively related to lake area, and positively related to dissolved organic carbon (DOC). All lakes were supersaturated in CO2 and the resulting carbon (C) flux to the atmosphere (150 to over 3000 mg C m2 day−1) was negatively related to lake area and positively to DOC concentration. CO2 fluxes were positively related to integrated water column respiration, suggesting a biological component in this flux. Both respiration and CO2 fluxes were strongly negatively related to years after the last fire in the basin, such that lakes in recently burnt basins had significantly higher C emissions, even after the influence of lake size was removed. No significant differences were found in nutrients, chlorophyll, and DOC between lakes in different basin types, suggesting that the fire‐induced watershed features influence other, more subtle aspects, such as the quality of the organic C reaching lakes. The fire‐induced enhancement of lake organic C mineralization and C emissions represents a long‐term impact that increases the overall C loss from the landscape as the result of fire, but which has never been included in current regional C budgets and future projections. The need to account for this additional fire‐induced C loss becomes critical in the face of predictions of increasing incidence of fire in the circumboreal landscape.
Local adaptation to higher temperatures reduces immigration success of genotypes from a warmer region in the water flea Daphnia
Authors: VAN DOORSLAER WENDY , VANOVERBEKE JOOST , DUVIVIER CATHY , ROUSSEAUX SARAH , JANSEN MIEKE , JANSEN BASTIAAN , FEUCHTMAYR HEIDRUN , ATKINSON DAVID , MOSS BRIAN , STOKS ROBBY , DE MEESTER LUC
Published: May 2009

There is growing awareness that microevolutionary dynamics may alter ecological processes. Rising temperatures under global change are expected to open windows for establishment of species and genotypes from warmer regions. Yet, microevolutionary tracking of temperature change by local populations may reduce establishment success of these immigrants. We exposed a UK population of the water flea, Daphnia magna, to two temperature regimes during a 1.5‐year experimental evolution trial, and subsequently compared competitive strength of non‐warm‐adapted and warm‐adapted D. magna in competition with French genotypes. Our results indicate that local microevolutionary responses to global warming may reduce establishment success of immigrant genotypes that are preadapted to warmer climate. Simulation modeling shows that microevolution results in a reduced likelihood and speed of displacement of local populations by immigrant genotypes under realistic immigrant/resident ratios. We conclude that local evolutionary dynamics may shift the relative impact of local and regional processes in response to global change.
Changes in species richness and composition in European acidic grasslands over the past 70 years: the contribution of cumulative atmospheric nitrogen deposition
Authors: DUPRÈ CECILIA , STEVENS CARLY J. , RANKE TRAUTE , BLEEKER ALBERT , PEPPLER‐LISBACH CORD , GOWING DAVID J. G. , DISE NANCY B. , DORLAND EDU , BOBBINK ROLAND , DIEKMANN MARTIN
Published: May 2009

Our study investigates the negative impact of nitrogen (N) deposition on species richness in acidic grasslands, based on a temporal comparison of vegetation data spanning a period of almost 70 years. We compiled a large data base of plots assigned to the Violion caninae grassland type, composed of managed, but unfertilized semi‐natural grasslands on nutrient‐poor, acidic soils. In total 1114 plots, mainly from Great Britain, the Netherlands and Germany, were compiled, dating back to 1939. Environmental site information included geographical and soil (mean Ellenberg values) variables as well as estimates of cumulative N and sulphur (S) deposition since 1939. Statistical analyses were carried out separately for the data subsets from the three regions. In all regions, the vegetation differentiation was mainly related to soil acidity and nutrient availability, as well as to the year of sampling and the cumulative amounts of N and S deposition. Plot‐species richness of vascular plants and bryophytes (analysed for Great Britain only) decreased with time and analyses suggest these are affected by various factors, notably soil pH, but also latitude and cumulative N deposition. The latter explained more of the variation in species number than the year of sampling and cumulative S deposition, which supports the interpretation that the decline in species richness is mainly caused by increasing N availability and less by altered management and soil acidification. For Great Britain and Germany, cumulative N deposition showed a strong negative relationship with several biodiversity measures, especially the proportion of dicots, whereas it was positively related to the proportion of grass species. In general, our results give temporal evidence for the negative effect of N deposition on species richness in semi‐natural vegetation.
Invasive insects impact forest carbon dynamics
Authors: CLARK KENNETH L. , SKOWRONSKI NICHOLAS , HOM JOHN
Published: May 2009

Invasive insects can impact ecosystem functioning by altering carbon, nutrient, and hydrologic cycles. In this study, we used eddy covariance to measure net CO2exchange with the atmosphere (NEE), and biometric measurements to characterize net ecosystem productivity (NEP) in oak‐ and pine‐dominated forests that were defoliated by Gypsy moth (Lymantria disparL.) in the New Jersey Pine Barrens. Three years of data were used to compare C dynamics; 2005 with minimal defoliation, 2006 with partial defoliation of the canopy and understory in a mixed stand, and 2007 with complete defoliation of an oak‐dominated stand, and partial defoliation of the mixed and pine‐dominated stands. Previous to defoliation in 2005, annual net CO2exchange (NEEyr) was estimated at −187, −137 and −204 g C m−2 yr−1at the oak‐, mixed‐, and pine‐dominated stands, respectively. Annual NEP estimated from biometric measurements was 108%, 100%, and 98% of NEEyrin 2005 for the oak‐, mixed‐, and pine‐dominated stands, respectively. Gypsy moth defoliation strongly reduced fluxes in 2006 and 2007 compared with 2005; NEEyrwas −122, +103, and −161 g C m−2 yr−1in 2006, and +293, +129, and −17 g C m−2 yr−1in 2007 at the oak‐, mixed‐, and pine‐dominated stands, respectively. At the landscape scale, Gypsy moths defoliated 20.2% of upland forests in 2007. We calculated that defoliation in these upland forests reduced NEEyrby 41%, with a 55% reduction in the heavily impacted oak‐dominated stands. ‘Transient’ disturbances such as insect defoliation, nonstand replacing wildfires, and prescribed burns are major factors controlling NEE across this landscape, and when integrated over time, may explain much of the patterning of aboveground biomass and forest floor mass in these upland forests.
Carbon sequestration in tropical forests and water: a critical look at the basis for commonly used generalizations
Authors: MALMER ANDERS , MURDIYARSO DANIEL , (SAMPURNO) BRUIJNZEEL L. A. , ILSTEDT ULRIK
Published: May 2009

Tree planting in the tropics is conducted for a number of reasons including carbon sequestration, but often competes with increasingly scarce water resources. The basics of forest and water relations are frequently said to be well understood but there is a pressing need to better understand and predict the hydrological effects of land‐use and climate change in the complex and dynamic landscapes of the tropics. This will remain elusive without the empirical data required to feed hydrological process models. It is argued that the current state of knowledge is confused by too broad a use of the terms ‘forest’ and ‘(af)forestation’, as well as by a bias towards using data generated mostly outside the tropics and for nondegraded soil conditions. Definitions of forest, afforestation and reforestation as used in the climate change community and their application by land and water managers need to be reconciled.
El Niño and dry season rainfall influence hostplant phenology and an annual butterfly migration from Neotropical wet to dry forests
Authors: SRYGLEY ROBERT B. , DUDLEY ROBERT , OLIVEIRA EVANDRO G. , AIZPRÚA RAFAEL , PELAEZ NICOLE Z. , RIVEROS ANDRE J.
Published: May 2009

We censused butterflies flying across the Panama Canal at Barro Colorado Island (BCI) for 16 years and butterfly hostplants for 8 years to address the question: What environmental factors influence the timing and magnitude of migrating Aphrissa statira butterflies? The peak migration date was earlier when the wet season began earlier and when soil moisture content in the dry season preceding the migration was higher. The peak migration date was also positively associated with peak leaf flushing of one hostplant (Callichlamys latifolia) but not another (Xylophragma seemannianum). The quantity of migrants was correlated with the El Niño Southern Oscillation, which influenced April soil moisture on BCI and total rainfall in the dry season. Both hostplant species responded to El Niño with greater leaf flushing, and the number of adults deriving from or laying eggs on those new leaves was greatest during El Niño years. The year 1993 was exceptional in that the number of butterflies migrating was lower than predicted by the El Niño event, yet the dry season was unusually wet for an El Niño year as well. Thus, dry season rainfall appears to be a primary driver of larval food production and population outbreaks for A. statira. Understanding how global climate cycles and local weather influence tropical insect migrations improves the predictability of ecological effects of climate change.
Reduced early growing season freezing resistance in alpine treeline plants under elevated atmospheric CO2
Authors: MARTIN MELISSA , GAVAZOV KONSTANTIN , KÖRNER CHRISTIAN , HÄTTENSCHWILER STEPHAN , RIXEN CHRISTIAN
Published: May 2009

The frequency of freezing events during the early growing season and the vulnerability to freezing of plants in European high‐altitude environments could increase under future atmospheric and climate change. We tested early growing season freezing sensitivity in 10 species, from four plant functional types (PFTs) spanning three plant growth forms (PGFs), from a long‐term in situ CO2 enrichment (566 vs. 370 ppm) and 2‐year soil warming (+4 K) experiment at treeline in the Swiss Alps (Stillberg, Davos). By additionally tracking plant phenology, we distinguished indirect phenology‐driven CO2 and warming effects from direct physiology‐related effects on freezing sensitivity. The freezing damage threshold (lethal temperature 50) under ambient conditions of the 10 treeline species spanned from −6.7±0.3 °C (Larix decidua) to −9.9±0.6 °C (Vaccinium gaultherioides). PFT, but not PGF, explained a significant amount of this interspecific variation. Long‐term exposure to elevated CO2 led to greater freezing sensitivity in multiple species but did not influence phenology, implying that physiological changes caused by CO2 enrichment were responsible for the effect. The elevated CO2 effect on freezing resistance was significant in leaves of Larix, Vaccinium myrtillus, and Gentiana punctata and marginally significant in leaves of Homogyne alpina and Avenella flexuosa. No significant CO2 effect was found in new shoots of Empetrum hermaphroditum or in leaves of Pinus uncinata, Leontodon helveticus, Melampyrum pratense, and V. gaultherioides. Soil warming led to advanced leaf expansion and reduced freezing resistance in V. myrtillus only, whereas Avenella showed greater freezing resistance when exposed to warming. No effect of soil warming was found in any of the other species. Effects of elevated CO2 and soil warming on freezing sensitivity were not consistent within PFTs or PGFs, suggesting that any future shifts in plant community composition due to increased damage from freezing events will likely occur at the individual species level.
Ecological feedbacks and the evolution of resistance
Authors: Duffy Meghan A. , Forde Samantha E.
Published: May 2009

1.
 The idea that parasites can affect host diversity is pervasive, and the possibility that parasites can increase host diversity is of particular interest. In this review, we focus on diversity in the resistance of hosts to their parasites, and on the different ways in which parasites can increase or decrease this resistance diversity.

2.
 Theoretically, parasites can exert many different types of selection on host populations, which each have consequences for host diversity. Specifically, theory predicts that parasites can exert negative frequency‐dependent selection (NFDS) and disruptive selection on resistance, both of which increase host diversity, as well as directional selection and stabilizing selection on resistance, both of which decrease host diversity.

3.
 Despite these theoretical predictions, most biologists think of only NFDS or directional selection for increased resistance in response to parasitism. Here, we present empirical support for all of these types of selection occurring in natural populations. Interestingly, several recent studies demonstrate that there is spatiotemporal variation in the type of selection that occurs (and, therefore, in the effects of parasitism on host diversity).

4.
 A key question that remains, then, is: What determines the type of parasite‐mediated selection that occurs? Theory demonstrates that the answer to this question lies, at least in part, with trade‐offs associated with resistance. Specifically, the type of evolution that occurs depends critically on the strength and shape of these trade‐offs. This, combined with empirical evidence for a strong effect of environment on the shape and strength of trade‐offs, may explain the observed spatiotemporal variation in parasite‐mediated selection.

5.
 We conclude that spatiotemporal variation in parasite‐driven evolution is likely to be common, and that this variation may be driven by ecological factors. We suggest that the feedback between ecological and evolutionary dynamics in host–parasite interactions is likely to be a productive area of research. In particular, studies addressing the role of ecological factors (e.g. productivity and predation regimes) in driving the outcome of parasite‐mediated selection on host populations are warranted. Such studies are necessary if we are to understand the mechanisms underlying the observed variation in the effects of parasites on host diversity.
Expression and characterization of the periplasmic cobalamin‐binding protein of Photobacterium damselae subsp. piscicida
Authors: Boiani R , Andreoni F , Serafini G , Bianconi I , Pierleoni R , Dominici S , Gorini F , Magnani M
Published: May 2009

Cobalamin (vitamin B12) is an essential cofactor in a variety of enzymatic reactions and most prokaryotes contain transport systems to import vitamin B12. A gene coding for a periplasmic cobalamin‐binding protein of Photobacterium damselae subsp. piscicida was identified by in silico analysis of sequences from a genomic library. The open reading frame was composed of 834 bp encoding a protein of 277 amino acids. The protein showed 61% identity with the vitamin B12‐binding protein precursor of P. profundum, 53% identity with the corresponding protein of Vibrio parahaemolyticus and 43% identity with the periplasmic binding protein BtuF of Escherichia coli. The expression of the native protein was investigated in P. damselae subsp. piscicida, but BtuF was weakly expressed under normal conditions. To characterize the BtuF of P. damselae subsp. piscicida, the recombinant protein was expressed with a C‐terminal His6‐tag and purified; the molecular weight was estimated to be approximately 30 kDa. The protein does not contain any free thiol group, consistent with the view that the two cysteine residues are involved in a disulphide bond. The purified BtuF binds cyanocobalamin with an affinity constant of 6 ± 2 μm.
Neutralizing antibody levels for protection against betanodavirus infection in sevenband grouper, Epinephelus septemfasciatus (Thunberg), immunized with an inactivated virus vaccine
Authors: Yamashita H , Mori K , Kuroda A , Nakai T
Published: May 2009

An inactivated betanodavirus, red‐spotted grouper nervous necrosis virus (RGNNV), is a vaccine candidate for viral nervous necrosis (VNN). The present study was conducted to examine inoculation doses of the vaccine and neutralizing antibody titre levels to protect fish against VNN. Young sevenband grouper, Epinephelus septemfasciatus, averaging 25.4 g, were immunized at 25 °C water temperature by a single intraperitoneal injection of formalin‐inactivated RGNNV. Fish immunized at vaccine doses of 108.5, 108.0, 107.5, 107.0 and 106.5 TCID50 per fish produced antibodies at mean titres of 1:907, 1:511, 1:259, 1:197 and 1:96, respectively, at 20 days post‐immunization (p.i.). Neutralizing antibodies were not detected in any control fish (titre <1:80). When fish were challenged with RGNNV (105.0 and 104.0 TCID50/fish) at 20 days p.i., cumulative mortalities of the fish groups immunized with 108.5, 108.0, 107.5 and 107.0 TCID50 per fish were significantly lower than those of the control group, and the relative percent survival values were higher than 60% in fish groups immunized with 107.5 TCID50 per fish or higher doses. However, no significant differences were found in mortality between the group immunized with 106.5 TCID50 per fish and the control group. From these results, it was deduced that the minimum effective inoculation dose of the vaccine is 107.0 TCID50 per fish and the minimum mean neutralizing antibody titre giving significant protection is approximately 1:200. This antibody titre level is a possible measure of vaccine efficacy against VNN in sevenband grouper, instead of a virus challenge test.
Molecular detection of three shrimp viruses and genetic variation of white spot syndrome virus in Hainan Province, China, in 2007
Authors: Tan Y , Xing Y , Zhang H , Feng Y , Zhou Y , Shi Z‐L
Published: May 2009

White spot syndrome virus (WSSV), Taura syndrome virus (TSV) and infectious hypodermal and haematopoietic necrosis virus (IHHNV) have been responsible for major pandemics affecting the shrimp farming industry. Shrimp samples were collected from eight farms in Hainan Province, China, during 2007 and analysed by polymerase chain reaction (PCR) or reverse transcriptase PCR methods to determine the prevalence of these viruses. From the eight sampling locations, only samples from one farm did not show any indication of infection with WSSV, TSV or IHHNV, while samples from one additional farm exhibited evidence of infection with TSV only. Surprisingly, evidence of co‐infection with TSV and IHHNV was found among samples at two farms while evidence of co‐infection with all three viruses (WSSV, TSV and IHHNV) was detected among shrimp samples at four farms. To further elucidate the molecular characteristics of WSSV in China, we further analysed genomic features of WSSV isolates based on the ORF23/24 variable region. From these data, we identified two novel WSSV strains which contain nucleotide deletions of 5657 and 11093 bp, respectively, when compared with the largest WSSV‐TW isolate.
Effects of the domestic use of a disclosing solution on the denture biofilm: a preliminary study
Authors: DE SOUZA R. F. , NASCIMENTO C. , REGIS R. R. , SILVA‐LOVATO C. H. , PARANHOS H. F. O.
Published: May 2009

Summary  To investigate the effect of the home use of a disclosing agent on the microbial composition of denture biofilm, by means of a cross‐over randomized clinical trial. Two interventions were tested during 7 days each: (I) oral and denture hygiene instructions and (II) instructions associated with the home use of a disclosing agent (1% neutral red). Eleven participants with visible biofilm deposits over their maxillary complete dentures were randomly assigned to one of the two sequences of interventions: (i) I followed by II, and (ii) II followed by I. A washout period of 7 days was established. After each intervention, samples of denture biofilm were evaluated by DNA checkerboard hybridization for the detection of Candida spp. and 17 bacterial species. Counts were low for all the tested species, and no significant difference was found between the tested interventions (Wilcoxon test, P > 0·05). The home use of a disclosing agent does not remarkably change the composition of denture biofilm.
Elongator mediates ABA responses, oxidative stress resistance and anthocyanin biosynthesis in Arabidopsis
Authors: Zhou Xiaofeng , Hua Deping , Chen Zhizhong , Zhou Zhengjun , Gong Zhizhong
Published: May 2009

Elongator is a histone acetyl‐transferase complex consisting of six subunits, and is highly conserved in eukaryotic organisms. Here, we isolated two novel mutants, elp2 and elp6, during a genetic screening for ABA‐hypersensitive Arabidopsis mutants. Map‐based cloning identified ELP2 and ELP6, which encode the orthologs of the yeast Elongator subunits, ELP2 and ELP6, respectively. Another Elongator subunit mutant, elp4/elo1, was obtained from the SALK T‐DNA collection. The elp1/abo1/elo2 mutant was isolated in a previous study. All four of the Elongator mutants had narrow leaves, reduced root growth, ABA hypersensitivity and an increased accumulation of anthocyanins. Mutations in the core subcomplex subunits ELP1/ABO1 and ELP2, but not in the accessory subcomplex subunits ELP4/ELO1 and ELP6, caused stomatal closing to be supersensitive to ABA. In addition, the four mutants were all more resistant than the wild type to oxidative stress produced by methyl viologen, and to CsCl. Gene expression analysis indicated that the four mutants had increased transcript levels of CAT3 under normal conditions, increased transcript levels of ZAT10 when treated with ABA and reduced transcript levels of MYBL2, which encodes a single‐repeat MYB protein, acting as a negative regulator of anthocyanin biosynthesis. Our results suggest that Elongator plays crucial roles in regulating plant responses to ABA, oxidative stress resistance and anthocyanin biosynthesis in Arabidopsis.
Functional characterization of a novel benzylisoquinoline O‐methyltransferase suggests its involvement in papaverine biosynthesis in opium poppy (Papaver somniferum L)
Authors: Pienkny Silke , Brandt Wolfgang , Schmidt Jürgen , Kramell Robert , Ziegler Jörg
Published: May 2009

The benzylisoquinoline alkaloids are a highly diverse group of about 2500 compounds which accumulate in a species‐specific manner. Despite the numerous compounds which could be identified, the biosynthetic pathways and the participating enzymes or cDNAs could be characterized only for a few selected members, whereas the biosynthesis of the majority of the compounds is still largely unknown. In an attempt to characterize additional biosynthetic steps at the molecular level, integration of alkaloid and transcript profiling across Papaver species was performed. This analysis showed high expression of an expressed sequence tag (EST) of unknown function only in Papaver somniferum varieties. After full‐length cloning of the open reading frame and sequence analysis, this EST could be classified as a member of the class II type O‐methyltransferase protein family. It was related to O‐methyltransferases from benzylisoquinoline biosynthesis, and the amino acid sequence showed 68% identical residues to norcoclaurine 6‐O‐methyltransferase. However, rather than methylating norcoclaurine, the recombinant protein methylated norreticuline at position seven with a Km of 44 μm using S‐adenosyl‐l‐methionine as a cofactor. Of all substrates tested, only norreticuline was converted. Even minor changes in the benzylisoquinoline backbone were not tolerated by the enzyme. Accordingly, the enzyme was named norreticuline 7–O‐methyltransferase (N7OMT). This enzyme represents a novel O‐methyltransferase in benzylisoquinoline metabolism. Expression analysis showed slightly increased expression of N7OMT in P. somniferum varieties containing papaverine, suggesting its involvement in the partially unknown biosynthesis of this pharmaceutically important compound.
Analysis of B function in legumes: PISTILLATA proteins do not require the PI motif for floral organ development in Medicago truncatula
Authors: Benlloch Reyes , Roque Edelín , Ferrándiz Cristina , Cosson Viviane , Caballero Teresa , Penmetsa Ramachandra V. , Beltrán José Pío , Cañas Luís Antonio , Ratet Pascal , Madueño Francisco
Published: May 2009

The B‐class gene PISTILLATA (PI) codes for a MADS‐box transcription factor required for floral organ identity in angiosperms. Unlike Arabidopsis, it has been suggested that legume PI genes contribute to a variety of processes, such as the development of floral organs, floral common petal–stamen primordia, complex leaves and N‐fixing root nodules. Another interesting feature of legume PI homologues is that some of them lack the highly conserved C‐terminal PI motif suggested to be crucial for function. Therefore, legume PI genes are useful for addressing controversial questions on the evolution of B‐class gene function, including how they may have diverged in both function and structure to affect different developmental processes. However, functional analysis of legume PI genes has been hampered because no mutation in any B‐class gene has been identified in legumes. Here we fill this gap by studying the PI function in the model legume species Medicago truncatula using mutant and RNAi approaches. Like other legume species, M. truncatula has two PI homologues. The expression of the two genes, MtPI and MtNGL9, has strongly diverged, suggesting differences in function. Our analyses show that these genes are required for petal and stamen identity, where MtPI appears to play a predominant role. However, they appear not to be required for development of the nodule, the common primordia or the complex leaf. Moreover, both M. truncatula PI homologues lack the PI motif, which indicates that the C‐terminal motif is not essential for PI activity.
Temporal and Spatial Distribution of Male Scent Marks in the Polygynous Greater Sac‐Winged Bat
Authors: Caspers Barbara A. , Voigt Christian C.
Published: May 2009

Scent marks are relatively long‐lived signals that can be perceived by conspecifics when the producer is absent. Therefore, it is often not obvious to whom the signal is directed. In daytime roosts of the polygynous greater sac‐winged bat, males scent mark territories with facial gland secretions. Territories are a valuable resource for males, as they offer exclusive courtship opportunities, which results in increased male reproductive success and, consequently, increased male–male competition over territories. The information encoded in male scent marks could, therefore, be either directed at females as part of an olfactory courtship display or at male competitors as part of territorial behaviour. We expected territorial males to scent mark in the morning, shortly before females return to the territory and close to female roosting sites, if scent marks are directed at females as part of the courtship display. And we expected harem males to scent mark at the territory boundaries, where male–male encounters are most likely to occur, if scent marks are directed at male competitors. We found that males marked more frequently in the afternoon, at a time when all females have already left the territory, and harem males marked at the territory boundaries and not inside their territory in the area where females roost. At boundaries males fan volatiles from specialised wing sacs towards competitors outside the territory. Scent marking of male Saccopteryx bilineata might therefore be congruent with the assessment‐hypothesis, which states that scent marks offer intruders the possibility to make an olfactory assessment of the territory owner without direct physical interaction. Thus, scent marks of male S. bilineata are most likely influenced by male–male competition and not by female choice.
Associations among symptoms of dieback in even‐aged stands of ash (Fraxinus excelsiorL.)
Authors: Skovsgaard J. P. , Thomsen I. M. , Skovgaard I. M. , Martinussen T.
Published: May 2009

The objective of this study was to establish statistically based associations among macroscopic symptoms of crown dieback, cankers due toChalara fraxinea, and symptoms caused by other pathogens and pests onFraxinus excelsior. A total of 454 trees were observed in two plots of a 15‐year‐old experimental stand. The symptoms included: (i) overall extent of crown dieback; (ii) dieback of upper parts of the crown; (iii) canker in upper parts of the crown; (iv) wilting foliage; (v) cankers and bark proliferations at the lower part of the stem; and (vi) discolouration at stump or stem base. The analysis suggested that the observed symptoms of crown dieback are caused by a primary disease. The macroscopic symptoms attributed to dieback and canker in the crown were strongly associated. Moreover, the disease was associated with symptoms ofArmillaria gallica, but no associations were found for symptoms ofNeonectria galligena,Pseudomonas syringaesubsp.savastanoipv.fraxini,Hylesinus fraxiniorH. variuswhen considered collectively. Dieback was more frequent on trees of average or below‐average size, suggesting that individual tree resistance decreased with decreasing growth potential or tree vigour. The extent of canker in the crown depended on site conditions and possibly on silvicultural practices. The development of phytosanitary prescriptions for silviculture should primarily be targeted towards young stands as these represent the most critical phases of stand development.
Multiple primary malignancies in Spanish patients with hepatocellular carcinoma: Analysis of a hospital‐based tumor registry
Authors: Fernández‐Ruiz Mario , Guerra‐Vales Juan‐Manuel , Castelbón‐Fernández Francisco‐Javier , Llenas‐García Jara , Caurcel‐Díaz Luis , Colina‐Ruizdelgado Francisco
Published: May 2009

Background and Aim:  Little is known about the etiological associations and clinical features of extrahepatic primary malignant (EHPM) neoplasms in subjects with hepatocellular carcinoma (HCC). The aim of this study was to characterize this phenomenon in a consecutive series of Spanish patients in order to define its natural history and influence on survival.

Methods:  A retrospective analysis of 245 patients with HCC during the period 1999–2003 was performed. Subjects identified with a second primary malignancy elsewhere constituted the EHPM group and were compared to patients with HCC alone.

Results:  Eighteen patients (7.3%) had one or two associated extrahepatic malignancies (mean age 67.7 ± 9.7 years); of these, 17 had double cancer and one patient, triple. Nine of the 19 EHPM occurred before HCC diagnosis. The associated cancers included five cases of colorectal carcinoma, four cases of head and neck carcinoma, three cases of genitourinary cancer, two cases of lymphoproliferative disorder, one lung carcinoma, one skin melanoma, one breast carcinoma, and two cancers of unknown origin. Age and sex distribution, etiology of underlying hepatopathy, and liver function tests did not differ significantly between both groups. There was no difference between the overall survival rates.

Conclusions:  EHPM is not rare among Spanish patients with HCC, although no specific clinicopathological features were detected in this population. Our results suggest that the association of another primary tumor with HCC does not imply a worse prognosis. The possibility of development of EHPM should be kept in mind when deciding on therapy and follow‐up of HCC.
Cytosine deaminase‐producing human mesenchymal stem cells mediate an antitumor effect in a mouse xenograft model
Authors: You Mi‐Hyeon , Kim Wang‐Joon , Shim Wooyoung , Lee Sang‐Rim , Lee Gwang , Choi Sangdun , Kim Dae‐Yong , Kim Yong Man , Kim Hyunsoo , Han Sang‐Uk
Published: May 2009

Background and Aim:  Stem cell transplantation offers potential gene therapy for brain tumors. However, this approach has received little attention as a treatment for gastrointestinal tumors. In the present study, we explored the possibility of human bone marrow‐derived mesenchymal stem cells (hMSC) producing cytosine deaminase (CD), followed by systemic 5‐fluorocytosine (5‐FC) administration, showing an antitumor effect on a mouse gastric cancer xenograft.

Methods:  We first explored the ability of hMSC, coated with fluorescent dye, to migrate to human gastric cancer MKN45 cells in vitro and in vivo. We then used hMSC in which a gene expressed the prodrug‐activating enzyme CD, which can convert the prodrug 5‐FC into the cytotoxic agent 5‐fluorouracil (5‐FU), and further investigated the potential of these cells to deliver the CD gene and to reduce tumor growth in nude mice. The migratory capacity of hMSC was confirmed by an in vitro migration assay, as well as in an in vivo model of nude mice bearing subcutaneous tumors of MKN45 cells when hMSC were injected.

Results:  The migration ability of hMSC towards MKN45 cells was confirmed by migration assay. Effective conversion of 5‐FC to 5‐FU by hMSC transfected with the CD gene (CD‐hMSC) showed therapeutic anticancer potential in a MKN45 cell co‐culture system, as confirmed by thin layer chromatography. Nude mice bearing MKN45 tumors were intravenously injected with CD‐hMSC, followed by systemic 5‐FC treatment (500 mg/kg/day) for 7 days. Tumor volumes and weights of mice injected with CD‐hMSC decreased significantly after treatment with 5‐FC. However, the 5‐FC‐treated group without CD‐hMSC injection showed neither a decrease in tumor volume nor bodyweight loss.

Conclusion:  The CD‐hMSC system showed anticancer therapeutic potential, and minimized the side‐effects of 5‐FU.
Characterization of hepatitis B virus reverse transcriptase sequences in Chinese treatment naive patients
Authors: Han Yue , Huang Li Hua , Liu Chuan Miao , Yang Shu , Li Juan , Lin Zhi Mei , Kong Xiao Fei , Yu De Min , Zhang Dong Hua , Jin Gen Di , Lu Zhi Meng , Gong Qi Ming , Zhang Xin Xin
Published: May 2009

Background and Aims:  The hepatitis B virus (HBV) reverse transcriptase (RT) plays an important role in viral replication. The aim of the present study was to characterize profiles of the RT region and to construct a database for further studies.

Methods:  Serum samples were obtained from 328 treatment naive patients chronically infected with HBV in five Chinese cities. Mutation status, genotypes and deep sequence analysis were carried out by amplifying and sequencing the RT region.

Results:  The base usage in the RT region differed at the mono‐ and dinucleotide level and thymidine dominated. The higher the variability of the strain was, the more it replicated. No significant clustering was found between our HBV RT sequences and those isolated 10 years ago (achieved from genebank). Nucleotide analogue resistance related mutants exist. The M204V/I mutation was found in 1.8% of the strains, 1.2% had L180M+ M204V/I, 0.6% had A181T/V, and only one had all three mutations. Minor strain mutants were found in 9.3% of the samples studied. The genotype B patients made up 36.6% (88.7% B2) and were mostly found in southern China, 63.4% (92.2% C2) were genotype C, and only one was genotype D. The average age of HBeAg positive genotype B patients was 29.5 ± 10.4 years, for genotype C it was 36.1 ± 10.9 (P < 0.001).

Conclusion:  Primarily antiviral resistance related mutant strains do exist in treatment naïve patients. Without antiviral pressure, HBV strains evolved at a normal speed. In depth sequence analysis implied that viral replication might be correlated with its variability, which needs to be further investigated.
Uridine diphosphate glucuronosyl transferase 1A1 promoter polymorphism is associated with choledocholithiasis in Taiwanese patients
Authors: Chu Cheng‐Hsin , Yang An‐Ming , Kao Jia‐Horng , Liu Chia‐Yuan , Chang Wen‐Hsiung , Yang Wei‐Shiung
Published: May 2009

Background and aims:  The gene product of the uridine diphosphate glucuronosyl transferase 1A1 (UGT1A1) is crucial to bilirubin metabolism. Mutations in this gene subsequently result in disease presented with unconjugated hyperbilirubinemia. A previous study showed that a TA‐repeat polymorphism in the promoter region of this gene might play a role in the metabolism of bilirubin. Whether this polymorphism might predispose choledocholithiasis is unclear.

Methods:  We recruited 32 patients who were diagnosed with pigment choledocholithiasis (common bile duct stones) by endoscopic retrograde cholangiopancreatography (ERCP) morphology and 107 population controls. The TA‐repeat in the UGT1A1 promoter was genotyped.

Results:  We found that among the 32 patients, 15 (46.9%) were wild type (A[TA]6TAA homozygous); 15 (46.9%) were a heterozygous variation (A[TA[6TAA/A[TA]7TAA) and 2 (6.2%) were a homozygous variation (A[TA]7TAA). Among the controls, 81 (75.7%) were wild type, 23 (21.5%) were a heterozygous variation and 3 (2.8%) were a homozygous variation. The genotype distribution was significantly different between patients and controls.

Conclusions:  The results suggest that the UGT1A1 promoter TA‐repeat polymorphism is associated with choledocholithiasis in Taiwanese patients.
The salivagram in severe cerebral palsy and able‐bodied adults
Authors: Baikie Gordon , Reddihough Dinah S , South Mike , Cook David J
Published: May 2009

Aims:  To test the hypothesis that the salivagram is frequently positive in those at high risk of aspiration but negative in individuals at low risk of aspiration lung disease.

Methods:  We studied 63 individuals with spastic quadriparetic cerebral palsy (CP), aged 14 months to 16 years (31 females and 32 males), and 20 able‐bodied young adult volunteers, aged 19–29 years (17 female and three males) using the salivagram. Twenty millilitres of 99mTc‐sulphur colloid in saline was instilled into the pharynx of each subject over 1 h. Aspiration was characterised by the appearance of activity in either lung field.

Results:  Aspiration was demonstrated by salivagram in 56% of children with CP (95% confidence interval 43% to 68%), compared to 0% of young adults (97.5% one‐sided confidence interval 0% to 17%). The difference in frequency is highly significant (P < 0.0006).

Conclusions:  The salivagram is frequently positive in those at high risk of aspiration, but frequently negative in those at low risk of aspiration. It may be a useful investigation for the demonstration of aspiration.
Impact of delivery mode and gestational age on haematological parameters in Taiwanese preterm infants
Authors: Wu Jun‐Ho , Chou Hung‐Chieh , Chen Pau‐Chung , Jeng Suh‐Fang , Chen Chien‐Yi , Tsao Po‐Nien , Hsieh Chia‐Jung , Huang Hsiu‐Min , Hsieh Wu‐Shiun
Published: May 2009

Aim:  Reference ranges of haematological parameters in preterm infants are limited. The aim of this study is to determine the reference values of haematological parameters in preterm infants in Taiwan, and to assess the impact of gestational age and mode of delivery on these parameters.

Method:  Medical records were retrospectively reviewed in preterm infants admitted to National Taiwan University Hospital from January 2001 to December 2004. The inclusion criteria included infants with <37 weeks of gestation who had blood sampling within 24 h of birth. The exclusion criteria included those with maternal history of antepartum haemorrhage, chorioamnionitis, fever, sepsis, preeclampsia and hypertension; and perinatal history of twin‐to‐twin transfusion syndrome, feto‐maternal transfusion, injury and infection.

Results:  Of 568 preterm infants with blood cell counts, 337 were available for analysis. There were trends of increase in red blood cell counts, haemoglobin levels and haematocrit values as gestation increased up to 34 weeks. In contrast, a trend of decrease was noted in mean corpuscular volume values. There was an initial trend of decrease in white blood cell counts and then increased after 31 weeks gestation. The platelet counts were essentially unchanged. Infants born by vaginal delivery generally had higher haematological parameters than those born by Caesarean section at different gestational ages except for mean corpuscular volume values.

Conclusions:  We established the reference ranges of haematological parameters in Taiwanese preterm infants. Health‐care professionals must be cautious in clinical application of the haematological values because of varying antenatal and perinatal risk factors.
The epidemiology of paediatric head injuries: Data from a referral centre in Victoria, Australia
Authors: Crowe Louise , Babl Franz , Anderson Vicki , Catroppa Cathy
Published: May 2009

Aim:  Currently, there are no population‐based or hospital‐based studies on the full spectrum of paediatric head injuries (HIs) in Australia. We set out to provide detailed information on the incidence rates, causes and clinical management of all severities of HI in children and adolescents at an Australian tertiary referral centre using emergency department (ED) and admission data as a basis for further investigations and prevention efforts.

Methods:  A retrospective chart review of all children aged 0–16 years who attended the Royal Childrens Hospital (RCH), Melbourne, following a HI in 2004 was used. The cases were identified using the International Classification of Diseases 10th revision codes, and all medical records were reviewed based on a piloted data form. Information was collected on demographics, injury factors and clinical management of HIs in the hospital setting.

Results:  Over the 12‐month period, there were 1115 children with an HI who attended the RCH ED, or were admitted. Ninety per cent were classified as mild, 8% as moderate and 3% as severe. Males and children under 3 years had the higher attendance rates. Falls, sports and motor vehicle accidents were the main HI causes. The main sport played (30%) when sustaining an HI was Australian rules football. Thirty‐two per cent of children were admitted, 67% of these with mild HI. Twenty‐one per cent had a radiology imaging study, most (67%) with a normal result.

Conclusions:  Many HI causes appear preventable, in particular, falls from heights in infants and sports safety. High rates of admission and radiology imaging of mild HI warrant further investigation.
The Pharmaceutical Benefits Scheme and implications for paediatric prescribing
Authors: Sinha Yashwant , Brien Jo‐anne E , Craig Jonathan C
Published: May 2009

Aims:  To evaluate the impact of the Pharmaceutical Benefits Advisory Committee (PBAC) decisions on access to medicines listed on the Pharmaceutical Benefits Scheme (PBS) for children.

Methods:  We analysed all public summary documents from PBAC meetings from July 2005 to November 2006 and compared these with the Therapeutic Goods Administration (TGA) recommendations for children for the same medicine. Main outcome measures stratified by age, the total number of medicines for specific indications (accepted and rejected) by therapeutic class; estimated cost to the PBS per annum for each medicine recommended for listing; comparison of TGA‐approved product information and PBS listing for recommended medicines.

Results:  Of the 102 medicines for specific indications considered by the PBAC, 7% (7/102) of submissions were for new paediatric indications. Most submissions (60%, 61/102) did not specify age for the PBS recommendation and were for conditions which only affect adults. Listings which specifically included children were more likely to have a positive PBAC recommendation. Of the six recommended medicines for children, four were estimated to cost between $10–30 million per year. There was fair concordance between PBS‐ and TGA‐approved product information for age (kappa 0.21) but in 46%, PBAC recommendations were for age‐unrestricted listing compared with adults‐only use in the TGA‐approved product information.

Conclusion:  Access to new subsidised medicines for children in Australia lags behind adults because most applications to the PBAC for new medicines are for conditions which only affect adults. PBS processes facilitate access for children to new medicines by avoiding age restrictions.