bluuebunny/crossref_metadata_2025 huggingface dataset:
Second‐hand smoking and carboxyhemoglobin levels in children: a prospective observational study
Authors: YEE BRANDEN E. , AHMED MOHAMMED I. , BRUGGE DOUG , FARRELL MAUREEN , LOZADA GUSTAVO , IDUPAGANTHI RAGHU , SCHUMANN ROMAN
Published: December 2009

Aim:  To establish baseline noninvasive carboxyhemoglobin (COHb) levels in children and determine the influence of exposure to environmental sources of carbon monoxide (CO), especially environmental tobacco smoke, on such levels.

Background:  Second‐hand smoking may be a risk factor for adverse outcomes following anesthesia and surgery in children (1) and may potentially be preventable.

Patients and Methods:  Parents and their children between the ages of 1–12 were enrolled on the day of elective surgery. The preoperative COHb levels of the children were assessed noninvasively using a CO‐Oximeter (Radical‐7 Rainbow SET Pulse CO‐Oximeter; Masimo, Irvine, CA, USA). The parents were asked to complete an environmental air‐quality questionnaire. The COHb levels were tabulated and correlated with responses to the survey in aggregate analysis. Statistical analyses were performed using the nonparametric Mann–Whitney and Kruskal–Wallis tests. P < 0.05 was statistically significant.

Results:  Two hundred children with their parents were enrolled. Children exposed to parental smoking had higher COHb levels than the children of nonsmoking controls. Higher COHb values were seen in the youngest children, ages 1–2, exposed to parental cigarette smoke. However, these trends did not reach statistical significance, and confidence intervals were wide.

Conclusions:  This study revealed interesting trends of COHb levels in children presenting for anesthesia and surgery. However, the COHb levels measured in our patients were close to the error margin of the device used in our study. An expected improvement in measurement technology may allow screening children for potential pulmonary perioperative risk factors in the future.
Anesthetic management of the hybrid stage 1 procedure for hypoplastic left heart syndrome (HLHS)
Authors: NAGUIB AYMEN N. , WINCH PETER , SCHWARTZ LAWRENCE , ISAACS JANET , RODEMAN ROBERTA , CHEATHAM JOHN P. , GALANTOWICZ MARK
Published: December 2009

Introduction:  Despite advances in the surgical and perioperative management of patients with hypoplastic left heart syndrome (HLHS), outcomes for this high‐risk group of patients remains suboptimal. The hybrid approach [bilateral pulmonary artery (PA) banding, ductal stenting, balloon atrial septostomy], is an emerging alternative therapy for the management of HLHS, which defers the risks of a major surgical repair until the infants are older. This article will describe our experience providing the anesthetic management of patients undergoing the hybrid procedure.

Methods:  After Institutional Review Board approval, we retrospectively reviewed the records of 77 patients who underwent the hybrid procedure as neonates between July 2002 and August 2008. We reviewed both the anesthetic and intensive care records.

Results:  The hybrid procedure was performed in 77 patients (31 female and 46 male). The average age of the patients was 11.8 days with an average weight of 2.98 kg. Fentanyl was used for analgesia at an average dose of 5.7 mcg·kg−1. The average increase in the systolic blood pressure after placement of the right and left PA bands was 11.3 mmHg. The average drop in the systemic saturation after placement of the bands was 7%, with an average postband and stent SaO2 of 82%. Twenty‐one patients received blood transfusion (27.3%) at an average dose of 43.5 ml (14.5 ml·kg−1). Forty patients received albumin during the case (51.9%) at an average dose of 23.2 ml (7.7 ml·kg−1). Seventeen patients arrived at the hybrid suite already intubated, and no attempt was made to extubate these patients at the end of the case. Thirty‐six patients were extubated at the end of the procedure, and a total of 64.9% of patients were extubated within the first 24 h postoperatively. Patients had notably stable hemodynamics throughout the first 24 h in the intensive care unit.

Discussion:  Patients undergoing the hybrid procedure have relatively stable intraoperative and early postoperative hemodynamics. The procedure is performed without cardiopulmonary bypass (CPB) and with minimal narcotic and anesthetic exposure. Patients typically do not require blood transfusions or inotropic support and are extubated at either the end of the procedure or within 24 h of ICU admission. In our experience, the anesthetic management of patients undergoing the hybrid procedure is straightforward and requires relatively few interventions when compared to traditional neonatal surgical repairs. Deferring the risks of anesthesia, CPB, hypothermic circulatory arrest, and prolonged postoperative sedation may yield developmental advantages to patients born with HLHS.
Effects of therapeutic suggestion in children undergoing general anesthesia: a randomized controlled trial
Authors: FORTIER MICHELLE A. , WEINBERG MEGAN , VITULANO LAWRENCE A. , CHORNEY JILL MACLAREN , MARTIN SARAH R. , KAIN ZEEV N.
Published: December 2009

Objectives and aim:  The goal of this randomized controlled trial was to examine the effect of intraoperative positive therapeutic suggestion on postoperative nausea and vomiting (PONV) in children undergoing general anesthesia and otolaryngological surgery.

Background:  Because of the high incidence of PONV following otolaryngological surgery and its negative impact on recovery, researchers have examined various nonpharmacological interventions to target this phenomenon. To date, the effectiveness of therapeutic suggestion has not been studied in children.

Methods:  Participants were 67 children undergoing tonsillectomy and adenoidectomy and their mothers. Children received a standardized anesthetic procedure and were randomly assigned to one of three interventions administered under general anesthesia: therapeutic suggestion, story (prosody control), or standard operating room noise. Children, parents, and healthcare personnel were blinded to group assignment. Nausea and vomiting were recorded in the postanesthesia care unit (PACU) and for the first 3 days at home.

Results:  Results demonstrated a decrease in nausea severity across the first 3 days, F2,49 = 10.37, P < 0.001, but no group differences in nausea severity in the PACU (F2,49 = 0.87, P = 0.43) or at home (F2,49 = 0.80, P = 0.46). There were also no group differences in vomiting episodes in the PACU (χ2 (2) = 1.25, P > 0.05) or at home (F2,49 = 1.59, P = 0.21).

Conclusions:  In this blinded controlled trial, therapeutic suggestion delivered intraoperatively did not impact children’s PONV. However, because this is the first study of this kind, replication may be needed.
A simultaneous test of trophic interaction models: which vegetation characteristic explains herbivore control over plant community mass?
Authors: Cronin James Patrick , Tonsor Stephen J. , Carson Walter P.
Published: December 2009

Ecology Letters (2010) 13: 202–212

Predicting herbivore control over plants (i.e. changes in plant mass due to herbivore damage) is a central goal of ecology. Progress has been limited, however, because the vegetation characteristics thought to influence herbivore control are naturally correlated and typically experimentally confounded. To address this problem, we defined eight conventional models that predict herbivore control over plant community mass, each model based on a different vegetation characteristic (i.e. host concentration, tissue nitrogen, growth rate, size, tolerance of herbivory or net primary productivity). We then used structural equation modelling to test each model against two field experiments. Our results clearly rejected all models except for a tolerance of herbivory mechanism; stems with greater access to limiting resources better tolerated herbivory, regardless of where herbivore activity was greatest. Consequently, herbivore reductions of plant community mass were greatest at low resource availability. This adds to evidence that herbivore activity poorly predicts herbivore control.
Taming the symbiont for coexistence: a host PGRP neutralizes a bacterial symbiont toxin
Authors: Troll Joshua V. , Bent Eric H. , Pacquette Nicholas , Wier Andrew M. , Goldman William E. , Silverman Neal , McFall‐Ngai Margaret J.
Published: December 2009

In horizontally transmitted mutualisms between marine animals and their bacterial partners, the host environment promotes the initial colonization by specific symbionts that it harvests from the surrounding bacterioplankton. Subsequently, the host must develop long‐term tolerance to immunogenic bacterial molecules, such as peptidoglycan and lipopolysaccaride derivatives. We describe the characterization of the activity of a host peptidoglycan recognition protein (EsPGRP2) during establishment of the symbiosis between the squid Euprymna scolopes and its luminous bacterial symbiont Vibrio fischeri. Using confocal immunocytochemistry, we localized EsPGRP2 to all epithelial surfaces of the animal, and determined that it is exported in association with mucus shedding. Most notably, EsPGRP2 was released by the crypt epithelia into the extracellular spaces housing the symbionts. This translocation occurred only after the symbionts had triggered host morphogenesis, a process that is induced by exposure to the peptidoglycan monomer tracheal cytotoxin (TCT), a bacterial ‘toxin’ that is constitutively exported by V. fischeri. Enzymatic analyses demonstrated that, like many described PGRPs, EsPGRP2 has a TCT‐degrading amidase activity. The timing of EsPGRP2 export into the crypts provides evidence that the host does not export this protein until after TCT induces morphogenesis, and thereafter EsPGRP2 is constantly present in the crypts ameliorating the effects of V. fischeri TCT.
The septal pore cap is an organelle that functions in vegetative growth and mushroom formation of the wood‐rot fungus Schizophyllum commune
Authors: Van Peer Arend F. , Wang Fengfeng , Van Driel Kenneth G. A. , De Jong Jan F. , Van Donselaar Elly G. , Müller Wally H. , Boekhout Teun , Lugones Luis G. , Wösten Han A. B.
Published: December 2009

Mushroom‐forming basidiomycetes colonize large areas in nature. Their hyphae are compartmentalized by perforated septa, which are usually covered by a septal pore cap (SPC). Here, we describe, for the first time, the composition and function of SPCs using the model system Schizophyllum commune. The SPC of S. commune was shown to consist of a proteinaceous matrix covered by a lipid membrane. The matrix was demonstrated to define the ultrastructure of the SPC and to consist of two main proteins, Spc14 and Spc33. Gene spc14 encodes a protein of 86 amino acids, which lacks known domain, signal or localization sequences. Gene spc33 encodes a 239 and a 340 amino acid variant. Both forms contain a predicted signal anchor that targets them to the ER. Immuno‐localization showed the presence of Spc33 in the SPC but not in ER. From this and previous reports it is concluded that the SPC is derived from this organelle. Inactivation of spc33 resulted in loss of SPCs and the inability to close septa. The latter may well explain why vegetative growth and mushroom formation were severely reduced in strains in which spc33 was inactivated.
Interactions between Nosema microspores and a neonicotinoid weaken honeybees (Apis mellifera)
Authors: Alaux Cédric , Brunet Jean‐Luc , Dussaubat Claudia , Mondet Fanny , Tchamitchan Sylvie , Cousin Marianne , Brillard Julien , Baldy Aurelie , Belzunces Luc P. , Le Conte Yves
Published: December 2009

Global pollinators, like honeybees, are declining in abundance and diversity, which can adversely affect natural ecosystems and agriculture. Therefore, we tested the current hypotheses describing honeybee losses as a multifactorial syndrome, by investigating integrative effects of an infectious organism and an insecticide on honeybee health. We demonstrated that the interaction between the microsporidia Nosema and a neonicotinoid (imidacloprid) significantly weakened honeybees. In the short term, the combination of both agents caused the highest individual mortality rates and energetic stress. By quantifying the strength of immunity at both the individual and social levels, we showed that neither the haemocyte number nor the phenoloxidase activity of individuals was affected by the different treatments. However, the activity of glucose oxidase, enabling bees to sterilize colony and brood food, was significantly decreased only by the combination of both factors compared with control, Nosema or imidacloprid groups, suggesting a synergistic interaction and in the long term a higher susceptibility of the colony to pathogens. This provides the first evidences that interaction between an infectious organism and a chemical can also threaten pollinators, interactions that are widely used to eliminate insect pests in integrative pest management.
Identification and characterization of the PhhR regulon in Pseudomonas putida
Authors: Herrera M. Carmen , Duque Estrella , Rodríguez‐Herva José J. , Fernández‐Escamilla Ana M. , Ramos Juan L.
Published: December 2009

Pseudomonas putida is a soil microorganism that utilizes aromatic amino acids present in root exudates as a nitrogen source. We have previously shown that the PhhR transcriptional regulator induces phhAB genes encoding a phenylalanine hydroxylase. In this study we show, using microarray assays and promoter fusions, that PhhR is a global regulator responsible for the activation of genes essential for phenylalanine degradation, phenylalanine homeostasis and other genes of unknown function. Recently, it has been shown that phenylalanine catabolism occurs through more than one pathway. One of these possible pathways involves the metabolism of phenylalanine via tyrosine, p‐hydroxyphenylpyruvate, and homogentisate. We identified two genes within this pathway that encode an acyl‐CoA transferase involved in the metabolism of acetoacetate. All genes in this pathway were induced in response to phenylalanine in a PhhR‐proficient background. The second potential degradative pathway involves the degradation of phenylalanine to produce phenylpyruvate, which seems to be degraded via phenylacetyl‐CoA. A number of mutants in the paa genes encoding phenylacetyl‐CoA degradation enzymes fail to grow on phenylpyruvate or phenylacetate, further supporting the existence of this second pathway. We found that the PhhR regulon also includes genes involved in the biosynthesis of aromatic amino acids that are repressed in the presence of phenylalanine, suggesting the possibility of feedback at the transcriptional level. In addition, we found that PhhR modulates the level of expression of the broad‐substrate‐specificity MexEF/OprN efflux pump. Expression from this pump is under the control of mexT gene product because phenylalanine‐dependent transcription from the mexE promoter does not occur in a mexT mutant background. These results place PhhR as an important regulator in the control of bacterial responses to aromatic amino acids.
RNA binding proteins mediate the ability of a fungus to adapt to the cold
Authors: Fang Weiguo , St. Leger Raymond J.
Published: December 2009

Little is known about how fungi adapt to chilling. In eubacteria, cold shock proteins (CSPs) facilitate translation by destabilizing RNA secondary structure. Animals and plants have homologous cold shock domains within proteins, and additional glycine‐rich RNA binding proteins (GRPs), but their role in stress resistance is poorly understood. In this study, we identified GRP homologues in diverse fungi. However, only Aspergillus clavatus and Metarhizium anisopliae possessed cold shock domains. Both M. anisopliae's small eubacteria‐like CSP (CRP1) and its GRP (CRP2) homologue were induced by cold. Disrupting either Crp1 or Crp2 greatly reduced metabolism and conidial germination rates at low temperatures, and decreased tolerance to freezing. However, while both Crp1 and Crp2 reduced freezing‐induced production of reactive oxygen species, only Crp1 protected cells against H2O2 and increased M. anisopliae's virulence to caterpillars. Unlike CRP2, CRP1 rescued the cold‐sensitive growth defects of an Escherichia coli CSP deletion mutant, and CRP1 also demonstrated transcription anti‐termination activity, so CRP1 can regulate transcription and translation at low temperature. Expressing either Crp1 or Crp2 in yeast increased metabolism at cold temperatures and Crp1 improved tolerance to freezing. Thus besides providing a model relevant to many biological systems, Crp1 and Crp2 have potential applications in biotechnology.
Metabolic responses of novel cellulolytic and saccharolytic agricultural soil Bacteria to oxygen
Authors: Schellenberger Stefanie , Kolb Steffen , Drake Harold L.
Published: December 2009

Cellulose is the most abundant biopolymer in terrestrial ecosystems and is degraded by microbial communities in soils. However, relatively little is known about the diversity and function of soil prokaryotes that might participate in the overall degradation of this biopolymer. The active cellulolytic and saccharolytic Bacteria in an agricultural soil were evaluated by 16S rRNA 13C‐based stable isotope probing. Cellulose, cellobiose and glucose were mineralized under oxic conditions in soil slurries to carbon dioxide. Under anoxic conditions, these substrates were converted primarily to acetate, butyrate, carbon dioxide, hydrogen and traces of propionate and iso‐butyrate; the production of these fermentation end‐products was concomitant with the apparent reduction of iron(III). [13C]‐cellulose was mainly degraded under oxic conditions by novel family‐level taxa of the Bacteroidetes and Chloroflexi, and a known family‐level taxon of Planctomycetes, whereas degradation under anoxic conditions was facilitated by the Kineosporiaceae (Actinobacteria) and cluster III Clostridiaceae and novel clusters within Bacteroidetes. Active aerobic sub‐communities in oxic [13C]‐cellobiose and [13C]‐glucose treatments were dominated by Intrasporangiaceae and Micrococcaceae (Actinobacteria) whereas active cluster I Clostridiaceae (Firmicutes) were prevalent in anoxic treatments. A very large number (i.e. 28) of the detected taxa did not closely affiliate with known families, and active Archaea were not detected in any of the treatments. These collective findings suggest that: (i) a large uncultured diversity of soil Bacteria was involved in the utilization of cellulose and products of its hydrolysis, (ii) the active saccharolytic community differed phylogenetically from the active cellulolytic community, (iii) oxygen availability impacted differentially on the activity of taxa and (iv) different redox guilds (e.g. fermenters and iron reducers) compete or interact during cellulose degradation in aerated soils.
Dual symbiosis of the vent shrimp Rimicaris exoculata with filamentous gamma‐ and epsilonproteobacteria at four Mid‐Atlantic Ridge hydrothermal vent fields
Authors: Petersen Jillian M. , Ramette Alban , Lott Christian , Cambon‐Bonavita Marie‐Anne , Zbinden Magali , Dubilier Nicole
Published: December 2009

The shrimp Rimicaris exoculata from hydrothermal vents on the Mid‐Atlantic Ridge (MAR) harbours bacterial epibionts on specialized appendages and the inner surfaces of its gill chamber. Using comparative 16S rRNA sequence analysis and fluorescence in situ hybridization (FISH), we examined the R. exoculata epibiosis from four vents sites along the known distribution range of the shrimp on the MAR. Our results show that R. exoculata lives in symbiosis with two types of filamentous epibionts. One belongs to the Epsilonproteobacteria, and was previously identified as the dominant symbiont of R. exoculata. The second is a novel gammaproteobacterial symbiont that belongs to a clade consisting exclusively of sequences from epibiotic bacteria of hydrothermal vent animals, with the filamentous sulfur oxidizer Leucothrix mucor as the closest free‐living relative. Both the epsilon‐ and the gammaproteobacterial symbionts dominated the R. exoculata epibiosis at all four MAR vent sites despite striking differences between vent fluid chemistry and distances between sites of up to 8500 km, indicating that the symbiosis is highly stable and specific. Phylogenetic analyses of two mitochondrial host genes showed little to no differences between hosts from the four vent sites. In contrast, there was significant spatial structuring of both the gamma‐ and the epsilonproteobacterial symbiont populations based on their 16S rRNA gene sequences that was correlated with geographic distance along the MAR. We hypothesize that biogeography and host–symbiont selectivity play a role in structuring the epibiosis of R. exoculata.
Streptomycetes contributing to atmospheric molecular hydrogen soil uptake are widespread and encode a putative high‐affinity [NiFe]‐hydrogenase
Authors: Constant Philippe , Chowdhury Soumitra Paul , Pratscher Jennifer , Conrad Ralf
Published: December 2009

Uptake of molecular hydrogen (H2) by soil is a biological reaction responsible for ∼80% of the global loss of atmospheric H2. Indirect evidence obtained over the last decades suggests that free soil hydrogenases with an unusually high affinity for H2 are carrying out the reaction. This assumption has recently been challenged by the isolation of Streptomyces sp. PCB7, displaying the high‐affinity H2 uptake activity previously attributed to free soil enzymes. While this finding suggests that actinobacteria could be responsible for atmospheric H2 soil uptake, the ecological importance of H2‐oxidizing streptomycetes remains to be investigated. Here, we show that high‐affinity H2 uptake activity is widespread among the streptomycetes. Among 14 streptomycetes strains isolated from temperate forest and agricultural soils, six exhibited a high‐affinity H2 uptake activity. The gene encoding the large subunit of a putative high‐affinity [NiFe]‐hydrogenase (hydB‐like gene sequence) was detected exclusively in the isolates exhibiting high‐affinity H2 uptake. Catalysed reporter deposition‐fluorescence in situ hybridization (CARD‐FISH) experiments targeting hydB‐like gene transcripts and H2 uptake assays performed with strain PCB7 suggested that streptomycetes spores catalysed the H2 uptake activity. Expression of the activity in term of biomass revealed that 106–107 H2‐oxidizing bacteria per gram of soil should be sufficient to explain in situ H2 uptake by soil. We propose that specialized H2‐oxidizing actinobacteria are responsible for the most important sink term in the atmospheric H2 budget.
Biogeography of wetland rice methanotrophs
Authors: Lüke Claudia , Krause Sascha , Cavigiolo Stefano , Greppi Diego , Lupotto Elisabetta , Frenzel Peter
Published: December 2009

We focused on the functional guild of methane oxidizing bacteria (MOB) as model organisms to get deeper insights into microbial biogeography. The pmoA gene was used as a functional and phylogenetic marker for MOB in two approaches: (i) a pmoA database (> 4000 sequences) was evaluated to obtain insights into MOB diversity in Italian rice paddies, and paddy fields worldwide. The results show a wide geographical distribution of pmoA genotypes that seem to be specifically adapted to paddy fields (e.g. Rice Paddy Cluster 1 and Rice Paddy Cluster 2). (ii) On the smaller geographical scale, we designed a factorial experiment including three different locations, two rice varieties and two habitats (soil and roots) within each of three rice fields. Multivariate analysis of terminal restriction fragment analysis profiles revealed different community patterns at the three field sites, located 10–20 km apart. Root samples were characterized by high abundance of type I MOB whereas the rice variety had no effect. With the agronomical practice being nearly identical, historical contingencies might be responsible for the field site differences. Considering a large reservoir of viable yet inactive MOB cells acting as a microbial seed bank, environmental conditions might have selected and activated a different subset at a time thereby shaping the community.
Transgenic maize lines with cell‐type specific expression of fluorescent proteins in plastids
Authors: Sattarzadeh Amir , Fuller Jonathan , Moguel Salvador , Wostrikoff Katia , Sato Shirley , Covshoff Sarah , Clemente Tom , Hanson Maureen , Stern David B.
Published: December 2009

Plastid number and morphology vary dramatically between cell types and at different developmental stages. Furthermore, in C4 plants such as maize, chloroplast ultrastructure and biochemical functions are specialized in mesophyll and bundle sheath cells, which differentiate acropetally from the proplastid form in the leaf base. To develop visible markers for maize plastids, we have created a series of stable transgenics expressing fluorescent proteins fused to either the maize ubiquitin promoter, the mesophyll‐specific phosphoenolpyruvate carboxylase (PepC) promoter, or the bundle sheath‐specific Rubisco small subunit 1 (RbcS) promoter. Multiple independent events were examined and revealed that maize codon‐optimized versions of YFP and GFP were particularly well expressed, and that expression was stably inherited. Plants carrying PepC promoter constructs exhibit YFP expression in mesophyll plastids and the RbcS promoter mediated expression in bundle sheath plastids. The PepC and RbcS promoter fusions also proved useful for identifying plastids in organs such as epidermis, silks, roots and trichomes. These tools will inform future plastid‐related studies of wild‐type and mutant maize plants and provide material from which different plastid types may be isolated.
A companion protease inhibitor for the protection of cytosol‐targeted recombinant proteins in plants
Authors: Goulet Charles , Benchabane Meriem , Anguenot Raphaël , Brunelle France , Khalf Moustafa , Michaud Dominique
Published: December 2009

We reported earlier the potential of tomato cathepsin D inhibitor (SlCDI) as an in‐built stabilizing agent for the protection of recombinant proteins in transgenic plant leaf crude extracts (Plant Biotechnol J.4, 359–368). Here we document the potential of SlCDI for the in situ protection of proteins in potato leaves. Total protein assays with control and SlCDI‐expressing potato lines indicated a positive impact of slcdi transgene expression on leaf protein content, with a mean relative increase of 35%–40% depending on the light regime. Out of ∼700 proteins detected on 2‐D gels, only 20 exhibited a significantly altered level on a protein‐specific basis, whereas most proteins were up‐regulated on a leaf fresh weight basis, albeit at variable rates. Quantitative reverse trancriptase‐PCR assays for rubisco activase showed similar transcript levels in leaves of test and control lines despite protein levels increased by two‐ to threefold in SlCDI‐expressing lines. These observations, along with the unrelated biological functions assigned to MS‐identified proteins up‐regulated in leaves and protease assays showing slightly increased proteasome activity in protein extracts of SlCDI‐expressing lines, suggest a general, proteasome‐independent protein stabilizing effect of SlCDI in planta. Transient expression assays with human α1‐antichymotrypsin also showed a stabilizing effect for SlCDI on heterologous proteins, leading to net levels of the human protein increased by ∼2.5‐fold in SlCDI‐expressing plants. These data illustrate, overall, the potential of SlCDI as an in vivo protein‐stabilizing agent in transgenic plant systems, useful to improve protein levels and recombinant protein accumulation.
Tubers from potato lines expressing a tomato Kunitz protease inhibitor are substantially equivalent to parental and transgenic controls
Authors: Khalf Moustafa , Goulet Charles , Vorster Juan , Brunelle France , Anguenot Raphaël , Fliss Ismaïl , Michaud Dominique
Published: December 2009

Recombinant protease inhibitors represent useful tools for the development of insect‐resistant transgenic crops, but questions have been raised in recent years about the impact of these proteins on endogenous proteases and chemical composition of derived food products. In this study, we performed a detailed compositional analysis of tubers from potato lines expressing the broad‐spectrum inhibitor of Ser and Asp proteases, tomato cathepsin D inhibitor (SlCDI), to detect possible unintended effects on tuber composition. A compositional analysis of key nutrients and toxic chemicals was carried out with tubers ofSlCDI‐expressing and control (comparator) lines, followed by a two‐dimensional gel electrophoresis (2‐DE) proteomic profiling of total and allergenic proteins to detect eventual effects at the proteome level. No significant differences were observed among control andSlCDI‐expressing lines for most chemicals assayed, in line with the very low abundance ofSlCDI in tubers. Likewise, proteins detected after 2‐DE showed no quantitative variation among the lines, except for a few proteins in some control and test lines, independent of slcditransgene expression. Components of the patatin storage protein complex and Kunitz protease inhibitors immunodetected after 2‐DE showed unaltered deposition patterns inSlCDI‐expressing lines, clearly suggesting a null impact of slcdion the intrinsic allergenic potential of potato tubers. These data suggest, overall, a null impact of slcdiexpression on tuber composition and substantial equivalence between comparator andSlCDI‐expressing tubers despite reported effects on leaf protein catabolism. They also illustrate the usefulness of proteomics as a tool to assess the authenticity of foods derived from novel‐generation transgenic plants.
Production of compact plants by overexpression of AtSHI in the ornamental Kalanchoë
Authors: Lütken Henrik , Jensen Lilli Sander , Topp Sine Hovbye , Mibus Heiko , Müller Renate , Rasmussen Søren K.
Published: December 2009

Growth retardation is an important breeding aim and an essential part of horticultural plant production. Here, the potential of transferring the Arabidopsis short internode (shi) mutant phenotype was explored by expressing the AtSHI gene in the popular ornamental plant Kalanchoë. A 35S‐AtSHI construct was produced and transferred into eight genetically different cultivars of Kalanchoë by Agrobacterium tumefaciens. The resulting transgenic plants showed dwarfing phenotypes like reduced plant height and diameter, and also more compact inflorescences, as a result of increased vegetative height. The shi phenotype was stable over more than five vegetative subcultivations. Compared with Arabidopsis, the ectopic expression of AtSHI in Kalanchoë showed several differences. None of the Kalanchoë SHI‐lines exhibited alterations in leaf colour or morphology, and most lines were not delayed in flowering. Moreover, continuous treatment of lines delayed in flowering with low concentrations of gibberellins completely restored the time of flowering. These features are very important as a delay in flowering would increase plant production costs significantly. The effect of expression controlled by the native Arabidopsis SHI promoter was also investigated in transgenic Kalanchoë and resulted in plants with a longer flowering period. Two AtSHI like genes were identified in Kalanchoë indicating a widespread presence of this transcription factor. These findings are important because they suggest that transformation with the AtSHI gene could be applied to several species as a tool for growth retardation, and that this approach could substitute the use of conventional chemical growth regulation in plant production.
Chloroplast‐derived vaccine antigens confer dual immunity against cholera and malaria by oral or injectable delivery
Authors: Davoodi‐Semiromi Abdoreza , Schreiber Melissa , Nalapalli Samson , Verma Dheeraj , Singh Nameirakpam D. , Banks Robert K. , Chakrabarti Debopam , Daniell Henry
Published: December 2009

Cholera and malaria are major diseases causing high mortality. The only licensed cholera vaccine is expensive; immunity is lost in children within 3 years and adults are not fully protected. No vaccine is yet available for malaria. Therefore, in this study, the cholera toxin‐B subunit (CTB) of Vibrio cholerae fused to malarial vaccine antigens apical membrane antigen‐1 (AMA1) and merozoite surface protein‐1 (MSP1) was expressed in lettuce and tobacco chloroplasts. Southern blot analysis confirmed homoplasmy and stable integration of transgenes. CTB‐AMA1 and CTB‐MSP1 fusion proteins accumulated up to 13.17% and 10.11% (total soluble protein, TSP) in tobacco and up to 7.3% and 6.1% (TSP) in lettuce, respectively. Nine groups of mice (n = 10/group) were immunized subcutaneously (SQV) or orally (ORV) with purified antigens or transplastomic tobacco leaves. Significant levels of antigen‐specific antibody titres of immunized mice completely inhibited proliferation of the malarial parasite and cross‐reacted with the native parasite proteins in immunoblots and immunofluorescence studies. Protection against cholera toxin challenge in both ORV (100%) and SQV (89%) mice correlated with CTB‐specific titres of intestinal, serum IgA and IgG1 in ORV and only IgG1 in SQV mice, but no other immunoglobulin. Increasing numbers of interleukin‐10+ T cell but not Foxp3+ regulatory T cells, suppression of interferon‐γ and absence of interleukin‐17 were observed in protected mice, suggesting that immunity is conferred via the Tr1/Th2 immune response. Dual immunity against two major infectious diseases provided by chloroplast‐derived vaccine antigens for long‐term (>300 days, 50% of mouse life span) offers a realistic platform for low cost vaccines and insight into mucosal and systemic immunity.
Twelve‐ to 14‐month‐old infants can predict single‐event probability with large set sizes
Authors: Denison Stephanie , Xu Fei
Published: December 2009

Previous research has revealed that infants can reason correctly about single‐event probabilities with small but not large set sizes (Bonatti, 2008; Teglas et al., 2007). The current study asks whether infants can make predictions regarding single‐event probability with large set sizes using a novel procedure. Infants completed two trials: A preference trial to determine whether they preferred pink or black lollipops and a test trial where infants saw two jars, one containing mostly pink lollipops and another containing mostly black lollipops. The experimenter removed one occluded lollipop from each jar and placed them in two separate opaque cups. Seventy‐eight percent of infants searched in the cup that contained a lollipop from the jar with a higher proportion of their preferred color object, significantly better than chance. Thus infants can reason about single‐event probabilities with large set sizes in a choice paradigm, and contrary to most findings in the infant literature, the prediction task used here appears a more sensitive measure than the standard looking‐time task.
Opportunistic infections and organ‐specific diseases in HIV‐1‐infected children: a cohort study (1990–2006)
Authors: Guillén S , García San Miguel L , Resino S , Bellón JM , González I , Jiménez de Ory S , Muñoz‐Fernández MA , Navarro ML , Gurbindo MD , De José MI , Mellado MJ , Martín‐Fontelos P , Gonzalez‐Tomé MI , Martinez J , Beceiro J , Roa MA , Ramos JT ,   on behalf of the Madrid Group for Research on Pediatric HIV Infection
Published: December 2009

Objectives

Highly active antiretroviral therapy (HAART) has dramatically changed the natural history of HIV infection in children, but there are few studies in the literature about the incidence of clinical manifestations after HAART in this population, compared with adults. The aim of this study was to describe the influence of the widespread use of HAART on the development of opportunistic infections and organ‐specific diseases in HIV‐infected children.

Methods

An observational study of a cohort of 366 vertically HIV‐infected children followed from 1990 to 2006 was carried out. According to the main antiretroviral protocol used, three calendar periods (CPs) were defined and compared: CP1 (1990–1996: no patients on HAART), CP2 (1997–1999: <60% on HAART) and CP3 (2000–2006: >60% on HAART).

Results

Children experienced a progressive increase in CD4 T cell count (P<0.05) and a decrease in HIV viral load from 1996 onwards (P<0.05). Similarly, rates of death, AIDS, opportunistic infections (bacteraemia, candidosis, cryptosporidiosis and bacterial pneumonia) and organ‐specific diseases (wasting syndrome, thrombocytopenia, cardiomyopathy, lymphoid interstitial pneumonia and HIV‐associated encephalopathy) were lower in CP2 and CP3 than in CP1.

Conclusions

This study provides evidence of improved clinical outcomes in HIV‐infected children over time and shows that mortality, AIDS, opportunistic infections and organ‐specific diseases declined as HAART was progressively instituted in this population.
Neutralizing antibodies, MxA expression and MMP‐9/TIMP‐1 ratio as markers of bioavailability of interferon‐beta treatment in multiple sclerosis patients: a two‐year follow‐up study
Authors: Garcia‐Montojo M. , Dominguez‐Mozo M. I. , De Las Heras V. , Bartolome M. , Garcia‐Martinez A. , Arroyo R. , Alvarez‐Lafuente R.
Published: December 2009

Background:  The objective of this study was to correlate the detection of neutralizing antibodies (NAbs) by the cytopathic effect (CPE) assay, with the expression of myxovirus resistance protein A (MxA), and the ratio between matrix metalloproteinase 9 (MMP‐9) and its tissular inhibitor (TIMP‐1), in order to evaluate their usefulness as markers of interferon beta (IFN‐beta) bioavailability.

Methods:  Pairs of blood and serum samples were collected from 50 patients with multiple sclerosis (MS) during 2 years of IFN‐beta treatment. Expression of MxA, MMP‐9 and TIMP‐1 were analysed by quantitative PCR, and NAbs were measured by CPE assay.

Results:  During the study, 60% of patients presented NAbs. The number of serum samples that were NAbs+ was significantly increased amongst patients with relapses (41/92 vs. 33/108, P = 0.04). With one serum sample and with a NAb titre >100 tenfold reduction unit (TRU), 66.7% of patients with MS suffered from relapses, 41.7% suffered from progression, and 75% was not an optimal clinical responder. We did not find any significant difference in MxA. We found that 62.5% of patients with MS patients whose ratio was increased twofold after 2 years suffered from relapses, 37.5% suffered from progression, and 68.7% was not an optimal clinical responder.

Conclusion:  The early detection of NAbs by CPE assay and the finding of only one serum sample with a NAb titre >100 TRU seem to be markers of low bioavailability of IFN‐beta, whilst a twofold decrease in the MMP‐9/TIMP‐1 ratio by quantitative PCR assay seems to be a marker of high bioavailability of IFN‐beta.
Hyperhomocysteinemia recurrence in levodopa‐treated Parkinson’s disease patients
Authors: Belcastro V. , Pierguidi L. , Castrioto A. , Menichetti C. , Gorgone G. , Ientile R. , Pisani F. , Rossi A. , Calabresi P. , Tambasco N.
Published: December 2009

Background:  Patients with Parkinson’s disease (PD) and chronically treated with L‐DOPA exhibit, in a percentage of 10–30%, supra‐physiological levels of plasma total homocysteinemia (tHcy). In this study, we have investigated, in a group of hyper‐homocysteinemic PD patients, the time of hyper‐tHcy recurrence after discontinuation of 1‐month folate supplementation given to normalize plasma tHcy levels.

Methods:  Plasma tHcy, cobalamin and folate were assayed before and after 1‐month folate supplementation (5 mg/day), and after 2 and 4 months after folate discontinuation in 29 PD patients (16M/13F, mean age 69.4 ± 6.9 years) stabilized on a mean L‐DOPA dose of 509.4 ± 312.1 mg/day.

Results:  After folate supplementation, plasma tHcy levels fell within the normal range in all patients. At the 2‐month control after folate discontinuation, plasma tHcy remained within physiological values in 25 out of 29 patients. Conversely, 4 months after folate discontinuation, all patients exhibited hyper‐tHcy.

Conclusions:  One‐month intake of 5 mg/day folate normalizes plasma tHcy levels in all hyper‐homocysteinemic PD patients. Following folate discontinuation, hyper‐tHcy recurs in all patients within 4 months. Knowledge of this time interval is useful to optimize pulses of folate therapy in hyper‐homocysteinemic patients with PD.
Psychological Acceptance Mediates the Impact of the Behaviour Problems of Children with Intellectual Disability on Fathers’ Psychological Adjustment
Authors: MacDonald Elaine E. , Hastings Richard P. , Fitzsimons Elaine
Published: December 2009

Background  Previous research with mothers of children with intellectual disabilities has shown that psychological acceptance is related to maternal psychological well‐being. The present research extended this line of enquiry to fathers and explored the potential for psychological acceptance to mediate the impact of children’s behaviour problems on paternal well‐being.

Method  Ninety‐nine fathers of 67 boys and 32 girls with intellectual disability, between 6 and 18 years of age, participated in a questionnaire study. Psychological acceptance of difficult/negative emotions and thoughts associated with interactions with the child with intellectual disability were measured alongside ratings of the child’s behaviour problems and paternal negative (stress, anxiety, depression) and positive (‘positive gain’) well‐being.

Results  Psychological acceptance was found to partially mediate the impact of child behaviour problems on paternal stress, anxiety, and depression. Acceptance was also a positive predictor of fathers’ perceptions of positive gain associated with raising their child with intellectual disability. However, it could not function as a potential mediator of positive gain in the present research because fathers’ ratings of their child’s behaviour problems were not associated with paternal positive gain.

Conclusions  Implications for practice include the potential of acceptance‐based interventions, and other psychological interventions targeting acceptance and avoidance processes (e.g. mindfulness‐based approaches), to positively affect paternal psychological adjustment.
Inequalities in Respite Service Provision: Insights from a National, Longitudinal Study of People with Intellectual Disabilities
Authors: McConkey Roy , Kelly Fionnola , Mannan Hasheem , Craig Sarah
Published: December 2009

Background  Respite or short breaks are frequently sought by parents and demand for them usually exceeds their availability.

Methods  Using data from a national database in Ireland of around 16 000 persons living with family carers, the availability of overnight respite provision was monitored over an 8‐year period along with the recorded needs for such services.

Results  Despite marked rises in the number of people receiving respite breaks resulting from increased government funding, there were marked inequalities in the availability of provision across the country. In recent years the inequalities decreased but still remained. The proportion of families requiring breaks also rose and a similar pattern of inequalities were found here too. Only a small proportion of families who had a recorded need in 1999 were receiving respite services 8 years later and over one third had a continuing need recorded.

Conclusion  This study highlighted some of the complexities in reducing inequalities in the provision of respite services and in identifying the need for them. It would be advantageous to develop more explicit criteria regarding the need for respite provision and to record the family’s preferences for the form this provision might take. These adjustments would add to the value of any national database as a service planning tool.
Predicting Mother/Father–Child Interactions: Parental Personality and Well‐being, Socioeconomic Variables and Child Disability Status
Authors: Glidden Laraine M. , Bamberger Katharine T. , Turek Kevin C. , Hill Kelli L.
Published: December 2009

Background  Child and parent characteristics as well as socioeconomic family variables can influence the quality of parent–child interactions.

Methods  Coders rated parent behaviour from a video‐taped 30‐min family interaction in 91 families rearing children who were either typically developing or had intellectual/developmental disabilities. In addition, mothers and fathers completed NEO‐FFI personality items as well as subjective well‐being ratings.

Results  Coder ratings were factor analysed, resulting in a four‐factor parent behaviour inventory. The disability status of the children did not predict ratings on three of the factors, but parents of children with disabilities were perceived as less negative on one factor than parents of typically developing children. Extraversion, occupational status, and subjective well‐being related to the child also predicted some parenting behaviours.

Conclusions  Parenting a child with intellectual/developmental disabilities did not result in identifiably adverse outcomes for either mothers or fathers as determined by both behavioural and self‐report measures.
Resveratrol reduces glutamate‐mediated monocyte chemotactic protein‐1 expression via inhibition of extracellular signal‐regulated kinase 1/2 pathway in rat hippocampal slice cultures
Authors: Lee Eun Ok , Park Hee Ju , Kang JiHee Lee , Kim Hye‐Sun , Chong Young Hae
Published: December 2009

J. Neurochem. (2010) 112, 1477–1488.

Published evidence has linked glutamate with the pathogenesis of Alzheimer’s disease (AD) and the up‐regulation of a variety of chemokines, including monocyte chemotactic protein‐1 (MCP‐1)/chemokine ligand 2, with AD‐associated pathological changes. In this study, we assessed the potential molecular basis for the role of glutamate in hippocampal inflammation by determining its effects on MCP‐1 induction. We also attempted to identify the mechanism by which resveratrol (trans‐3,5,4′‐trihydroxystilbene), a polyphenolic phytostilbene, modulates the expression of MCP‐1 in the glutamate‐stimulated hippocampus. An ex vivo study using rat hippocampal slices demonstrated a time‐ and dose‐dependent increase in MCP‐1 release from glutamate‐exposed hippocampus. This increase was accompanied by enhanced MCP‐1 gene expression via the activation of the MEK/extracellular signal‐regulated kinase (ERK) pathway and interleukin‐1β (IL‐1β) expression. The inhibition of the MEK/ERK pathway with SL327, which is capable of crossing the blood‐brain barrier, nearly abolished the observed glutamate‐induced effects. Furthermore, anti‐IL‐1β antibodies suppressed the glutamate‐induced expression of MCP‐1 mRNA and protein, whereas an isotype‐matched antibody exerted only minimal effects. It is worthy of note that resveratrol, to a similar degree as SL327, down‐regulated glutamate‐induced IL‐1β expression and reduced the expression of MCP‐1 mRNA and protein release via the inactivation of ERK1/2. These results indicate that the activation of the MEK/ERK pathway and the consequent IL‐1β expression are essential for glutamate‐stimulated MCP‐1 production in the hippocampus. Additionally, our data reveal an anti‐inflammatory mechanism of resveratrol involving the inactivation of the ERK1/2 pathway in the hippocampus, which is linked principally to AD‐associated cognitive dysfunction.