bluuebunny/crossref_metadata_2025 huggingface dataset:
NepA is a structural cell wall protein involved in maintenance of spore dormancy in Streptomyces coelicolor
Authors: De Jong Wouter , Manteca Angel , Sanchez Jesus , Bucca Giselda , Smith Colin P. , Dijkhuizen Lubbert , Claessen Dennis , Wösten Han A. B.
Published: February 2009

Streptomycetes have a complex morphogenetic programme culminating in the formation of aerial hyphae that develop into chains of spores. After spore dispersal, environmental signals trigger dormant spores to germinate to establish a new colony. We here compared whole genome expression of a wild‐type colony of Streptomyces coelicolor forming aerial hyphae and spores with that of the chp null mutant that forms few aerial structures. This revealed that expression of 244 genes was significantly altered, among which genes known to be involved in development. One of the genes that was no longer expressed in the ΔchpABCDEFGH mutant was nepA, which was previously shown to be expressed in a compartment connecting the substrate mycelium with the sporulating parts of the aerial mycelium. We here show that expression is also detected in developing spore chains, where NepA is secreted to end up as a highly insoluble protein in the cell wall. Germination of spores of a nepA deletion mutant was faster and more synchronous, resulting in colonies with an accelerated morphogenetic programme. Crucially, spores of the ΔnepA mutant also germinated in water, unlike those of the wild‐type strain. Taken together, NepA is the first bacterial structural cell wall protein that is important for maintenance of spore dormancy under unfavourable environmental conditions.
Structural and functional abnormalities in lungs in patients with achalasia
Authors: Makharia G. K. , Seith A. , Sharma S. K. , Sinha A. , Goswami P. , Aggarwal A. , Puri K. , Sreenivas V.
Published: February 2009

Abstract  Dilatation and oesophageal body aperistalsis in achalasia can lead to stasis which in turn can induce repeated microaspiration. It is therefore conceivable that patients with achalasia may also have abnormalities in lungs secondary to repeated episodes of microaspiration. There is a lack of systematic study on involvement of lungs in patients with achalasia. Thirty patients with achalasia underwent pulmonary function tests (spirometry, and carbon mono‐oxide diffusion capacity) and high resolution computerized tomography (HRCT) of the chest. The mean age of patients and mean duration of disease were 33.5 ± 10.9 years and 28.1 ± 27.3 months respectively. Regurgitation was present in 22 (73.3%) of them. Respiratory symptoms in them were dry cough in 17 (56.6%), and chest pain in 18 (60%). The oesophagus was dilated in 26 (86.6%) and 13 (43.3%) had residue in oesophagus. Sixteen (53.3%) patients had either anatomical changes as seen on HRCT or functional changes as observed on pulmonary function tests. Of those with functional abnormalities, five (16.6%) and one (3.3%) had restrictive and obstructive airways disease respectively. While evidence of tracheo‐bronchial compression by dilated oesophagus was present in eight (26.6%), 10 (33.3%) patients had parenchymal lung disease [nodular opacities in five (16.6%), ground glass appearance six (20%), patchy pulmonary fibrosis five (16.6%), air trapping two (6.6%), consolidation and bronchiectasis one (3.3%) each]. There was a significant association between presence of regurgitation and dilatation of oesophagus (P = 0.032). More than half (53.3%) of patients with achalasia have structural and/or functional abnormalities in lungs.
Influence of naloxone on rectal sensorimotor function in health
Authors: Geeraerts B. , V. Oudenhove L. , Vos R. , Karamanolis G. , Tack J.
Published: February 2009

Abstract  Abnormal rectal motor physiology and visceral hypersensitivity are implicated in the pathogenesis of irritable bowel syndrome. Endogenous opioids are involved in both the regulation of gut motility and the processing of sensory information. Our aim was to study the effect of suppression of endogenous opioid function by naloxone on rectal sensorimotor function in health. Eighteen healthy subjects participated in a rectal barostat study. Sensorimotor function was evaluated during two consecutive stepwise distensions separated by 30 min of basal tone recording, and with perception scoring on a 0–6 graded scale. Naloxone was administered, after 15 min of basal tone measurements, as an intravenous bolus (0.4 mg), followed by continuous infusion (20 μg kg−1 h−1) in a placebo‐controlled, single‐blinded and randomized fashion. Naloxone did not alter rectal sensitivity. Comparison of visual analogue scale scores between naloxone and saline did not reveal altered intensities of pain or discomfort. Compared to the baseline distension, a significant adaptive increase in compliance occurred during the second distension after saline (7.8 ± 0.7 vs 11.0 ± 0.6 mL mmHg−1, P = 0.0016). This dynamic change in rectal compliance did not occur after naloxone administration (8.8 ± 0.7 vs 10.1 ± 0.8 mL mmHg−1, ns). Low intensity tonic distension induced a rectal adaptive relaxation, which was absent after naloxone. Naloxone does not alter rectal sensitivity but abolishes rectal adaptation in response to repeated balloon distention. These observations suggest that the endogenous opioid system is involved in control of rectal tone rather than rectal sensitivity.
Multiple rapid swallowing: a complementary test during standard oesophageal manometry
Authors: Fornari F. , Bravi I. , Penagini R. , Tack J. , Sifrim D.
Published: February 2009

Abstract  Multiple rapid swallowing (MRS) stimulates neural inhibition resulting in abolition of contractions in the oesophageal body (OB) and complete lower oesophageal sphincter (LOS) relaxation which is followed by peristalsis and LOS contraction. The aim of this study was to evaluate the yield of MRS to detect abnormalities in inhibitory or excitatory oesophageal mechanisms in patients with oesophageal symptoms and either normal standard manometry or ineffective oesophageal motility (IOM). MRS (five water swallows, 2 mL, separated by 2–3 s) was evaluated in 23 healthy subjects, 109 symptomatic patients with normal standard sleeve manometry and in 48 patients with IOM. Healthy subjects had complete inhibition of OB motility during MRS and a strong motor response after MRS, i.e. amplitude of OB contractions in the oesophageal body and LOS tone being higher than after single swallows. Almost 70% of patients with oesophageal symptoms and normal manometry had abnormal MRS, mainly consistent on inability to increase amplitude of OB contractions after MRS. Nearly, half of the patients with IOM were able to normalize OB contractions after MRS. MRS is a simple complementary test that can be added to standard oesophageal manometry. Two‐thirds of patients with normal manometry show abnormal MRS that could potentially underlie their symptoms. A normal response to MRS in patients with severe IOM might be used to predict response to prokinetic treatment.
Duration of control of two soilborne pathogens following incorporation of above‐ and below‐ground residues of Brassica juncea into soil
Authors: Motisi N. , Montfort F. , Doré T. , Romillac N. , Lucas P.
Published: February 2009

The persistence of control of primary infections caused by two soilborne fungal plant pathogens, Rhizoctonia solani and Gaeumannomyces graminis var. tritici, following the incorporation of above‐ground parts (AP), below‐ground parts (BP) or both (AP+BP) of Brassica juncea into soil was examined through an experiment in controlled conditions. Control was quantified by measuring disease incidence in bioassays where inoculum was introduced at different dates after the incorporation of plant residues. All types of residue showed an unexpected long‐term persistence that lasted at least 13 days, while the predominant glucosinolates contained in AP (20·9 µmol sinigrin g−1 dry matter) and BP (2·3 µmol gluconasturtiin g−1 dry matter) were hydrolysed in less than 3 days. Temporal trends in the efficacy of the residues behaved mostly in a quadratic manner, suggesting that the noxious effect of residues may be attributable to the release of isothiocyanates during the first days following incorporation, but that other mechanisms are most likely to contribute to lasting persistence. Across all treatments, AP and AP+BP suppressed R. solani by 54 and 63%, respectively, and G. graminis var. tritici by 40 and 40%, respectively, compared with controls. While BP did not cause any additional detectable effect when combined with AP, they had a significant effect when incorporated alone (approximately 20% suppression of both species), suggesting the existence of a complex interaction between these two types of residue.
POT1‐independent single‐strand telomeric DNA binding activities in Brassicaceae
Authors: Shakirov Eugene V. , McKnight Thomas D. , Shippen Dorothy E.
Published: February 2009

Telomeres define the ends of linear eukaryotic chromosomes and are required for genome maintenance and continued cell proliferation. The extreme ends of telomeres terminate in a single‐strand protrusion, termed the G‐overhang, which, in vertebrates and fission yeast, is bound by evolutionarily conserved members of the POT1 (protection of telomeres) protein family. Unlike most other model organisms, the flowering plantArabidopsis thalianaencodes two divergent POT1‐like proteins. Here we show that the single‐strand telomeric DNA binding activity present inA. thaliananuclear extracts is not dependent on POT1a or POT1b proteins. Furthermore, in contrast to POT1 proteins from yeast and vertebrates, recombinant POT1a and POT1b proteins fromA. thaliana, and from two additional Brassicaceae species,Arabidopsis lyrataandBrassica oleracea(cauliflower), fail to bind single‐strand telomeric DNAin vitrounder the conditions tested. Finally, although we detected four single‐strand telomeric DNA binding activities in nuclear extracts fromB. oleracea, partial purification and DNA cross‐linking analysis of these complexes identified proteins that are smaller than the predicted sizes of BoPOT1a or BoPOT1b. Taken together, these data suggest that POT1 proteins are not the major single‐strand telomeric DNA binding activities inA. thalianaand its close relatives, underscoring the remarkable functional divergence of POT1 proteins from plants and other eukaryotes.
The effector protein Avr2 of the xylem‐colonizing fungus Fusarium oxysporum activates the tomato resistance protein I‐2 intracellularly
Authors: Houterman Petra M. , Ma Lisong , Van Ooijen Gerben , De Vroomen Marianne J. , Cornelissen Ben J.C. , Takken Frank L.W. , Rep Martijn
Published: February 2009

To promote host colonization, many plant pathogens secrete effector proteins that either suppress or counteract host defences. However, when these effectors are recognized by the host’s innate immune system, they trigger resistance rather than promoting virulence. Effectors are therefore key molecules in determining disease susceptibility or resistance. We show here that Avr2, secreted by the vascular wilt fungus Fusarium oxysporum f. sp. lycopersici (Fol), shows both activities: it is required for full virulence in a susceptible host and also triggers resistance in tomato plants carrying the resistance gene I‐2. Point mutations in AVR2, causing single amino acid changes, are associated with I‐2‐breakingFol strains. These point mutations prevent recognition by I‐2, both in tomato and when both genes are co‐expressed in leaves of Nicotiana benthamiana. Fol strains carrying the Avr2 variants are equally virulent, showing that virulence and avirulence functions can be uncoupled. Although Avr2 is secreted into the xylem sap when Fol colonizes tomato, the Avr2 protein can be recognized intracellularly by I‐2, implying uptake by host cells.
Interplay between SCARECROW, GA and LIKE HETEROCHROMATIN PROTEIN 1 in ground tissue patterning in the Arabidopsis root
Authors: Cui Hongchang , Benfey Philip N.
Published: February 2009

Regulated cell division is critical for the development of multi‐cellular organisms. In the Arabidopsis root, SCARECROW (SCR) is required for the first cell division, but represses the subsequent, longitudinal asymmetric cell divisions that generate the two cell types of the ground tissue – cortex and endodermis. To elucidate the molecular basis of the role of SCR in ground tissue patterning, we screened for SCR‐interacting proteins using the yeast two‐hybrid method. A number of putative SCR‐interacting proteins were identified, among them LIKE HETEROCHROMATIN PROTEIN 1 (LHP1). In lhp1 mutants, a second longitudinal asymmetric cell division occurs in the ground tissue earlier than in wild‐type plants. Similar to the scr mutant, this premature middle cortex phenotype is suppressed by the phytohormone gibberellin (GA). We provide evidence that the N‐terminal domain of SCR is required for the interaction between SCR and LHP1 as well as with other interacting partners, and that this domain is essential for repression of asymmetric cell divisions. Consistent with a role for GA in cortex proliferation, mutants of key GA signaling components produce a middle cortex precociously. Intriguingly, we found that the spindly (spy) mutant has a similar middle cortex phenotype. As SPY homologs in animals physically interact with histone deacetylase, we examined the role of histone deacetylation in middle cortex formation. We show that inhibition of histone deacetylase activity causes premature middle cortex formation in wild‐type roots. Together, these results suggest that epigenetic regulation is probably the common basis for SCR and GA activity in cortex cell proliferation.
Cytosolic activity of SPINDLY implies the existence of a DELLA‐independent gibberellin‐response pathway
Authors: Maymon Inbar , Greenboim‐Wainberg Yaarit , Sagiv Sivan , Kieber Joseph J. , Moshelion Menachem , Olszewski Neil , Weiss David
Published: February 2009

Specific plant developmental processes are modulated by cross‐talk between gibberellin (GA)‐ and cytokinin‐response pathways. Coordination of the two pathways involves the O‐linked N‐acetylglucosamine transferase SPINDLY (SPY) that suppresses GA signaling and promotes cytokinin responses in Arabidopsis. Although SPY is a nucleocytoplasmic protein, its site of action and targets are unknown. Several studies have suggested that SPY acts in the nucleus, where it modifies nuclear components such as the DELLA proteins to regulate signaling networks. Using chimeric GFP–SPY fused to a nuclear‐export signal or to a glucocorticoid receptor, we show that cytosolic SPY promotes cytokinin responses and suppresses GA signaling. In contrast, nuclear‐localized GFP–SPY failed to complement the spy mutation. To examine whether modulation of cytokinin activity by GA and spy is mediated by the nuclear DELLA proteins, cytokinin responses were studied in double and quadruple della mutants lacking the activities of REPRESSOR OF GA1‐3 (RGA) and GA‐INSENSITIVE (GAI) or RGA, GAI, RGA Like1 (RGL1) and RGL2. Unlike spy, the della mutants were cytokinin‐sensitive. Moreover, when GA was applied to a cytokinin‐treated quadruple della mutant it was able to suppress various cytokinin responses. These results suggest that cytosolic SPY and GA regulate cytokinin responses via a DELLA‐independent pathway(s).
PLASTOCHRON3/GOLIATH encodes a glutamate carboxypeptidase required for proper development in rice
Authors: Kawakatsu Taiji , Taramino Graziana , Itoh Jun‐Ichi , Allen Justin , Sato Yutaka , Hong Soon‐Kwan , Yule Ryan , Nagasawa Nobuhiro , Kojima Mikiko , Kusaba Makoto , Sakakibara Hitoshi , Sakai Hajime , Nagato Yasuo
Published: February 2009

Most aerial parts of the plant body are products of the continuous activity of the shoot apical meristem (SAM). Leaves are the major component of the aerial plant body, and their temporal and spatial distribution mainly determines shoot architecture. Here we report the identification of the rice gene PLASTOCHRON3 (PLA3)/GOLIATH (GO) that regulates various developmental processes including the rate of leaf initiation (the plastochron). PLA3/GO encodes a glutamate carboxypeptidase, which is thought to catabolize small acidic peptides and produce small signaling molecules. pla3 exhibits similar phenotypes to pla1 and pla2– a shortened plastochron, precocious leaf maturation and rachis branch‐to‐shoot conversion in the reproductive phase. However, in contrast to pla1 and pla2, pla3 showed pleiotropic phenotypes including enlarged embryo, seed vivipary, defects in SAM maintenance and aberrant leaf morphology. Consistent with these pleiotropic phenotypes, PLA3 is expressed in the whole plant body, and is involved in plant hormone homeostasis. Double mutant analysis revealed that PLA1, PLA2 and PLA3 are regulated independently but function redundantly. Our results suggest that PLA3 modulates various signaling pathways associated with a number of developmental processes.
Molecular and reverse genetic characterization of NUCLEOSOME ASSEMBLY PROTEIN1 (NAP1) genes unravels their function in transcription and nucleotide excision repair in Arabidopsis thaliana
Authors: Liu Ziqiang , Zhu Yan , Gao Juan , Yu Fang , Dong Aiwu , Shen Wen‐Hui
Published: February 2009

Compared with the well‐studied biochemical function of NUCLEOSOME ASSEMBLY PROTEIN1 (NAP1) as a histone chaperone in nucleosome assembly/disassembly, the physiological roles of NAP1 remain largely uncharacterized. Here, we define the NAP1 gene family members in Arabidopsis, examine their molecular properties, and use reverse genetics to characterize their biological roles. We show that the four AtNAP1‐group proteins can form homodimers and heterodimers, can bind histone H2A, and are localized abundantly in the cytoplasm and weakly in the nucleus at steady state. AtNAP1;4 differs from the others by showing inhibitor‐sensitive nucleocytoplasmic shuttling and tissue‐specific expression, restricted to root segments and pollen grains. The other three AtNAP1 genes are ubiquitously expressed in plants and the AtNAP1;3 protein is detected as the major isoform in seedlings. We show that disruption of the AtNAP1‐group genes does not affect normal plant growth under our laboratory conditions. Interestingly, two allelic triple mutants, Atnap1;1‐1 Atnap1;2‐1 Atnap1;3‐1 and Atnap1;1‐1 Atnap1;2‐1 Atnap1;3‐2, exhibit perturbed genome transcription, and show hypersensitivity to DNA damage caused by UV‐C irradiation. We show that AtNAP1;3 binds chromatin, with enrichment at some genes involved in the nucleotide excision repair (NER) pathway, and that the expression of these genes is downregulated in the triple mutants. Taken together, our results highlight conserved and isoform‐specific properties of AtNAP1 proteins, and unravel their function in the NER pathway of DNA damage repair.
The plant adherin AtSCC2 is required for embryogenesis and sister‐chromatid cohesion during meiosis in Arabidopsis
Authors: Sebastian Jose , Ravi Maruthachalam , Andreuzza Sebastien , Panoli Aneesh P. , Marimuthu Mohan P. A. , Siddiqi Imran
Published: February 2009

Adherin plays an important role in loading the cohesin complex onto chromosomes, and is essential for the establishment of sister‐chromatid cohesion. We have identified and analyzed the Arabidopsis adherin homolog AtSCC2. Interestingly, the sequence analysis of AtSCC2 and of other putative plant adherin homologs revealed the presence of a PHD finger, which is not found in their fungal and animal counterparts. AtSCC2 is identical to EMB2773, and mutants show early embryo lethality and formation of giant endosperm nuclei. A role for AtSCC2 in sister‐chromatid cohesion was established by using conditional RNAi and examining meiotic chromosome organization. AtSCC2‐RNAi lines showed sterility, arising from the following defects in meiotic chromosome organization: failure of homologous pairing, loss of sister‐chromatid cohesion, mixed segregation of chromosomes and chromosome fragmentation. The mutant phenotype, which included defects in chromosome organization and cohesion in prophase I, is distinct from that of the Arabidopsis cohesin mutant Atrec8, which retains centromere cohesion up to anaphase I. Immunostaining experiments revealed the aberrant distribution of the cohesin subunit AtSCC3 on chromosomes, and defects in chromosomal axis formation, in the meiocytes of AtSCC2‐RNAi lines. These results demonstrate a role for AtSCC2 in sister‐chromatid cohesion and centromere organization, and show that the machinery responsible for the establishment of cohesion is conserved in plants.
PCR and direct agglutination as Leishmania infection markers among healthy Nepalese subjects living in areas endemic for Kala‐Azar
Authors: Bhattarai Narayan Raj , Van der Auwera Gert , Khanal Basudha , De Doncker Simonne , Rijal Suman , Das Murari Lal , Uranw Surendra , Ostyn Bart , Praet Nicolas , Speybroeck Niko , Picado Albert , Davies Clive , Boelaert Marleen , Dujardin Jean‐Claude
Published: February 2009

Objective  To compare a PCR assay and direct agglutination test (DAT) for the detection of potential markers of Leishmania infection in 231 healthy subjects living in a kala‐azar endemic focus of Nepal.

Methods  The sample was composed of 184 (80%) persons without any known history of KA and not living in the same house as known kala‐azar cases (HNK), 24 (10%) Healthy Household Contacts (HHC) and 23 (10%) past kala‐azar cases which had been successfully treated (HPK).

Results  PCR and DAT positivity scores were, respectively: HNK, 17.6% and 5.6%; HHC, 12.5% and 20.8%; HPK, 26.1% and 95.7%. The ratio PCR‐positives/DAT‐positives was significantly higher in HNK (ratio = 3.1) than in HHC (ratio = 0.6, P = 0.036) and in HPK (ratio = 0.2, P = 0.012). The ratio PCR‐positives/DAT‐positives did not significantly differ between HHC (ratio = 0.6) and HPK (ratio = 0.2, P = 0.473). The positive agreement index between PCR and DAT in HNK was 5%; in HHC, 0%; in HPK, 43%.

Conclusions  Our study highlights the specific character of PCR and DAT for the exploration of Leishmania asymptomatic infections. PCR is probably more informative for very recent infections among HNK, while DAT provides more information among HHC and HPK, a feature likely related to the power of serology to track less recent infections.
Control of pyrethroid‐resistant Anopheles gambiae and Culex quinquefasciatus mosquitoes with chlorfenapyr in Benin
Authors: N’Guessan Raphael , Boko Pelagie , Odjo Abiba , Knols Bart , Akogbeto Martin , Rowland Mark
Published: February 2009

Objective  To compare the efficacy of chlorfenapyr applied on mosquito nets and as an indoor residual spray against populations of Anopheles gambiae and Culex quinquefasciatus in an area of Benin that shows problematic levels of pyrethroid resistance.

Method  Eight‐week trial conducted in experimental huts.

Results  Indoor residual spraying killed 82.9% of An. gambiae overall (mean mortality: 79.5%) compared to 53.5% overall (mean mortality: 61.7%) in the hut containing the lower dosed ITN. Analysis of data on a fortnightly basis showed high levels of mosquito mortality and blood‐feeding inhibition during the first few weeks after treatment. Control of C. quinquefasciatus by the IRS and ITN interventions showed a similar trend to that of An. gambiae and though the average level of mortality was lower it was still much higher than with pyrethroid treatments against this population. Chlorfenapyr’s reputation for being rather slow acting was evident particularly at lower dosages. The treatments showed no evidence of excito‐repellent activity in this trial.

Conclusion  Chlorfenapyr has the potential to control pyrethroid resistant populations of A. gambiae. There is a need to develop long‐lasting formulations of chlorfenapyr to prolong its residual life on nets and sprayed surfaces. On nets it could be combined with a contact irritant pyrethroid to give improved protection against mosquito biting while killing pyrethroid‐resistant mosquitoes that come into contact with the net.
Rapid malaria diagnostic tests vs. clinical management of malaria in rural Burkina Faso: safety and effect on clinical decisions. A randomized trial
Authors: Bisoffi Zeno , Sirima Bienvenu Sodiomon , Angheben Andrea , Lodesani Claudia , Gobbi Federico , Tinto Halidou , Van den Ende Jef
Published: February 2009

Objectives  To assess if the clinical outcome of patients treated after performing a Rapid Diagnostic Test for malaria (RDT) is at least equivalent to that of controls (treated presumptively without test) and to determine the impact of the introduction of a malaria RDT on clinical decisions.

Methods  Randomized, multi‐centre, open clinical trial in two arms in 2006 at the end of the dry and of the rainy season in 10 peripheral health centres in Burkina Faso: one arm with use of RDT before treatment decision, one arm managed clinically. Primary endpoint: persistence of fever at day 4. Secondary endpoints: frequency of malaria treatment and of antibiotic treatment.

Results  A total of 852 febrile patients were recruited in the dry season and 1317 febrile patients in the rainy season, and randomized either to be submitted to RDT (P_RTD) or to be managed presumptively (P_CLIN). In both seasons, no significant difference was found between the two randomized groups in the frequency of antimalarial treatment, nor of antibiotic prescription. In the dry season, 80.8% and 79.8% of patients with a negative RDT were nevertheless diagnosed and treated for malaria, and so were 85.0% and 82.6% negative patients in the rainy season. In the rainy season only, both diagnosis and treatment of other conditions were significantly less frequent in RDT positive vs. negative patients (48.3%vs. 61.4% and 46.2%vs. 59.9%, P = 0.00 and 0.00, respectively).

Conclusion  Our study was inconclusive on RDT safety (clinical outcome in the two randomized groups), because of an exceedingly and unexpectedly low compliance with the negative test result. Further research is needed on best strategies to promote adherence and on the safety of a test based strategy compared with the current, presumptive treatment strategy.
Successful oral tolerance induction in severe peanut allergy
Authors: Clark A. T. , Islam S. , King Y. , Deighton J. , Anagnostou K. , Ewan P. W.
Published: February 2009

Background:  Peanut allergy is common, potentially severe and rarely resolves causing impaired quality of life. No disease‐modifying treatment exists and there is therefore a need to develop a therapeutic intervention.

Aims of the study:  The aim of this study was to investigate whether peanut oral immunotherapy (OIT) can induce clinical tolerance to peanut protein.

Methods:  Four peanut‐allergic children underwent OIT. Preintervention oral challenges were performed to confirm clinical allergy and define the amount of protein required to cause a reaction (dose thresholds). OIT was then administered as daily doses of peanut flour increasing from 5 to 800 mg of protein with 2‐weekly dose increases. After 6 further weeks of treatment, the oral challenge was repeated to define change in dose threshold and subjects continued daily treatment.

Results:  Preintervention challenges confirmed peanut allergy and revealed dose thresholds of 5–50 mg (1/40–1/4 of a whole peanut); one subject had anaphylaxis during challenge and required adrenaline injection. All subjects tolerated immunotherapy updosing to 800 mg protein and i.m. adrenaline was not required. Each subject tolerated at least 10 whole peanuts (approximately 2.38 g protein) in postintervention challenges, an increase in dose threshold of at least 48‐, 49‐, 55‐ and 478‐fold for the four subjects.

Conclusions:  We demonstrated a substantial increase in dose threshold after OIT in all subjects, including the subject with proven anaphylaxis. OIT was well tolerated and conferred protection against at least 10 peanuts, more than is likely to be encountered during accidental ingestion.
Serum interleukin‐17 levels are related to clinical severity in allergic rhinitis
Authors: Ciprandi G. , De Amici M. , Murdaca G. , Fenoglio D. , Ricciardolo F. , Marseglia G. , Tosca M.
Published: February 2009

Background:  T helper (Th)‐17 cells are a subset of T helper lymphocytes that exert regulatory activities. Recently, it has been reported that serum interleukin (IL)‐17 levels are high in the most severe cases of birch allergy studied outside the pollen season.

Objective:  The aim of this study was to investigate a possible relationship between serum IL‐17 levels and clinical parameters in patients with allergic rhinitis studied during the pollen season.

Methods:  In all, 56 patients with persistent pollen‐induced allergic rhinitis were evaluated during the pollen season. Serum IL‐17 levels were evaluated by enzyme‐linked immunosorbent assay. Symptoms were assessed by visual analogue scale, drug use was monitored and peripheral eosinophils were counted.

Results:  Serum IL‐17 levels were significantly related to clinical symptoms, drug use and peripheral eosinophil counts (P = 0.0001 for all).

Conclusion:  This study provides evidence that serum IL‐17 level assessment might be considered to classify allergy severity.
ECMO in ARDS: a long‐term follow‐up study regarding pulmonary morphology and function and health‐related quality of life
Authors: LINDÉN V. B. , LIDEGRAN M. K. , FRISÉN G. , DAHLGREN P. , FRENCKNER B. P. , LARSEN F.
Published: February 2009

Background: A high survival rate can be achieved in patients with severe acute respiratory distress syndrome (ARDS) using extracorporeal membrane oxygenation (ECMO). The technique and the costs are, however, debated and follow‐up studies in survivors are few. The aim of this study was to evaluate long‐term pulmonary health after ECMO and severe ARDS.

Methods: Twenty‐one long‐term survivors of severe ARDS and ECMO were studied in a follow‐up program including high‐resolution computed tomography (HRCT) of the lungs, extensive pulmonary function tests, pulmonary scintigraphy and the pulmonary disease‐specific St George's Respiratory Questionnaire (SGRQ).

Results: The majority of patients had residual lung parenchymal changes on HRCT suggestive of fibrosis, but the extension of morphologic abnormalities was limited and without the typical anterior localization presumed to indicate ventilator‐associated lung injury. Pulmonary function tests revealed good restitution with mean values in the lower normal range, while T½ for outwash of inhaled isotope was abnormal in all patients consistent with subclinical obstructivity. Most patients had reduced health‐related quality of life (HRQoL), according to the SGRQ, but were stating less respiratory symptoms than conventionally treated ARDS patients in previous studies. The majority were integrated in normal work.

Conclusion: The majority of ECMO‐treated ARDS patients have good physical and social functioning. However, lung parenchymal changes on HRCT suggestive of fibrosis and minor pulmonary function abnormalities remain common and can be detected more than 1 year after ECMO. Furthermore, most patients experience a reduction in HRQoL due to the pulmonary sequelae.
Systemic inflammation in the brain‐dead organ donor
Authors: BARKLIN A.
Published: February 2009

Brain death itself impairs organ function in the potential donor, thereby limiting the number of suitable organs for transplantation. In addition, graft survival of kidneys obtained from brain‐dead (BD) donors is inferior to that of kidneys obtained from living donors. Experimental studies confirm an inferior graft survival for the heart, liver and lungs from BD compared with living donors. The mechanism underlying the deteriorating effect of brain death on the organs has not yet been fully established. We know that brain death triggers massive circulatory, hormonal and metabolic changes. Moreover, the past 10 years have produced evidence that brain death is associated with a systemic inflammatory response. However, it remains uncertain whether the inflammation is induced by brain death itself or by events before and after becoming BD. The purpose of this study is to discuss the risk factors associated with brain death in general and the inflammatory response in the organs in particular. Special attention will be paid to the heart, lung, liver and kidney and evidence will be presented from clinical and experimental studies.
Comparison between intubation and the laryngeal mask airway in moderately obese adults
Authors: ZOREMBA M. , AUST H. , EBERHART L. , BRAUNECKER S. , WULF H.
Published: February 2009

Background: Obesity is a well‐established risk factor for perioperative pulmonary complications. Anaesthetic drugs and the effect of obesity on respiratory mechanics are responsible for these pathophysiological changes, but tracheal intubation with muscle relaxation may also contribute. This study evaluates the influence of airway management, i.e. intubation vs. laryngeal mask airway (LMA), on postoperative lung volumes and arterial oxygen saturation in the early postoperative period.

Methods: We prospectively studied 134 moderately obese patients (BMI 30) undergoing minor peripheral surgery. They were randomly assigned to orotracheal intubation or LMA during general anaesthesia with mechanical ventilation. Premedication, general anaesthesia and respiratory settings were standardized. While breathing air, we measured arterial oxygen saturation by pulse oximetry. Inspiratory and expiratory lung function was measured preoperatively (baseline) and at 10 min, 0.5, 2 and 24 h after extubation, with the patient supine, in a 30° head‐up position. The two groups were compared using repeated‐measure analysis of variance (ANOVA) and t‐test analysis. Statistical significance was considered to be P<0.05.

Results: Postoperative pulmonary mechanical function was significantly reduced in both groups compared with preoperative values. However, within the first 24 h, lung function tests and oxygen saturation were significantly better in the LMA group (P<0.001; ANOVA).

Conclusions: In moderately obese patients undergoing minor surgery, use of the LMA may be preferable to orotracheal intubation with respect to postoperative saturation and lung function.
Coevolution between crossbills and black pine: the importance of competitors, forest area and resource stability
Authors: BENKMAN C. W. , PARCHMAN T. L.
Published: February 2009

Studies of predator‐prey interactions have found that geographically structured coevolution has played an important role in the adaptive diversification of crossbills (Loxia spp.). We extend those studies by considering common crossbills (L. curvirostra) in the Mediterranean where they rely on seeds in the cones of black pine (Pinus nigra). On the continent, where tree squirrels (Sciurus vulgaris) are present, enhanced defenses against crossbills were most evident in larger areas of pine forest. On islands in the absence of tree squirrels, crossbills and black pine have coevolved in a predator‐prey arms race on Cyprus but not Corsica. In contrast to other conifers that island endemic crossbills rely upon, black pine does not hold seeds in its cones year round. Consequently, key to the strong crossbill–pine interaction on Cyprus is likely the presence of an alternative conifer that provides seeds during early summer when black pine seeds are scarce.
Immunogenic males: a genome‐wide analysis of reproduction and the cost of mating in Drosophila melanogaster females
Authors: INNOCENTI P. , MORROW E. H.
Published: February 2009

In Drosophila melanogaster, mating radically transforms female physiology and behaviour. Post‐mating responses include an increase in the oviposition rate, a reduction in female receptivity and an activation of the immune system. The fitness consequences of mating are similarly dramatic – females must mate once in order to produce fertile eggs, but additional matings have a clear negative effect. Previously, microarrays have been used to examine gene expression of females differing in their reproductive status with the aim of identifying genes influenced by mating. However, as only virgin and single mated females were compared, transcriptional changes associated with reproduction (under natural selection) and male‐induced effects (possibly under sexually antagonistic selection) cannot be disentangled. We partitioned these fundamentally different effects by instead examining the expression profiles of virgin, single mated and double mated females. We found substantial effects relating to reproduction and further effects that are only attributable to mating itself. Immune response genes dominate this male‐induced effect indicating that the cost of mating may be due partly to this system’s activation. We propose that both sexually antagonistic and natural selection have been important in the evolution of the innate immunity genes, thereby contributing to the sexual dimorphism and rapid evolution at these loci.
Monitoring viral incidence rates: tools for the implementation of European Union regulations
Authors: Janssen M. P. , Cator E. A. , Van Der Poel C. L. , Schaasberg W. P. , Bonsel G. J. , Van Hout B. A.
Published: February 2009

Background and Objectives European legislation requires manufacturers of plasma products to report epidemiological data on human immunodeficiency virus, hepatitis B virus and hepatitis C virus in donor populations. The incidence rates of such infections are directly related to the risk of infection transmission. We propose two statistical tests to evaluate these incidence rates.

Materials and Methods Infection data of the four Dutch blood collection centres from 2003 through 2006 were analysed. For transversal comparison of centres and detection of increased incidence rates, a new statistical test was developed (outlier test). For longitudinal detection of trends in incidence rates, a generic test for trend is proposed. The power and risk of non‐detection are evaluated for both tests.

Results Application of the outlier test did not reveal any significantly increased incidence rates among centres in The Netherlands. The test for trend showed no significant increase in incidence rates in individual centres, but on national level a statistically significant increase in hepatitis C virus incidence was observed (P‐value of 0·01).

Conclusion The proposed tests allow signalling of outlier centres and trends in incidence rates both at individual centre and at national levels. Graphical support and the use of as much relevant historical data as possible is recommended. The statistical tests described are generic and can be applied by any blood establishment and plasma fractionation institute.
Influence of beach morphodynamics on the bivalve Donax hanleyanus and Mesodesma mactroides populations in Argentina
Authors: Thompson Gustavo A. , Sánchez De Bock Mariana F.
Published: February 2009

Five exposed microtidal sandy beaches of North‐eastern Buenos Aires (Argentina) were analysed to evaluate the influence of coastal dynamics on the abundance of the intertidal clams Donax hanleyanus and Mesodesma mactroides over a 1‐year period. The physical characterisation of each beach was performed using 15 environmental variables and six morphodynamic indices. The population densities of both clam species were estimated by the number of individuals per strip‐transect. Analysis of similarities revealed a significant multivariate environmental difference among beaches, which could be aggregated into a northern and a southern group. The former group was characterised by an intermediate morphodynamic state and by having, in general, high density values for both clam species. The latter was composed of beaches with an intermediate reflective state and showed, in general, low density values. Beach Index and Area showed the lowest temporal variation. Total density values, as well as density values of recruits and adults of D. hanleyanus and M. mactroides did not follow a north–south latitudinal gradient. Spatial and temporal fluctuations of density values were best explained by changes in individual physical variables rather than by changes in composite indices.
Molecular mapping of genic male‐sterile genes ms15, ms5 and ms6 in tetraploid cotton
Authors: Chen D. , Ding Y. , Guo W. , Zhang T.
Published: February 2009

Two genic male sterile (GMS) lines, Lang‐A conditioned by ms15 and Zhongkang‐A conditioned by ms5ms6 duplicate recessive genes in Gossypium hirsutum L., were chosen to map GMS genes. These two lines were crossed with Gossypium barbadense cv. ‘Hai7124’ to produce segregating populations. The ms15 gene was mapped on chromosome 12, and was flanked by two simple sequence repeat (SSR) markers, NAU2176 and NAU1278, with a genetic distance of 0.8 and 1.9 cM respectively. The ms5 and ms6 genes were mapped to one pair of homoeologous chromosomes, ms5 on chromosome 12 flanked by three SSR markers, NAU3561, NAU2176 and NAU2096, with genetic distances of 1.4, 1.8 and 1.8 cM, respectively, and ms6 on chromosome 26 flanked by two SSR markers, BNL1227 and NAU460, with a genetic distance of 1.4 and 1.7 cM respectively. These tightly linked markers with the ms15, ms5 and ms6 genes can be used in the marker‐assisted selection among segregating populations in a breeding programme, and provide the foundation for gene isolation by map‐based cloning for these three genes.
Combining ability of maize inbred lines containing genes from Zea diploperennis for resistance to Striga hermonthica (Del.) Benth
Authors: Yallou C. G. , Menkir A. , Adetimirin V. O. , Kling J. G.
Published: February 2009

Maize (Zea mays) is a staple food crop in sub‐Saharan Africa, but its production is threatened by Striga hermonthica (Del.) Benth. Transfer of resistance genes from wild relatives may increase resistance to S. hermonthica in tropical maize. The objective of this study was to determine the combining ability of resistance to S. hermonthica among lines containing Zea diploperennis and tropical germplasm. Forty‐five diallel crosses of 10 inbred lines were evaluated in an alpha‐lattice design with and without artificial Striga infestation at two locations each in the Republic of Benin and Nigeria for 3 years. Results of analyses showed that only general combining ability (GCA) mean square was significant (P = 0.01) for number of emerged Striga plants (NESPP), while both GCA and specific combining ability (SCA) mean squares were significant for host damage score (HDS) and grain yield under Striga infestation. The ratio of GCA to SCA mean squares for the three traits varied from 3.5 to 57.5. Although GCA × environment interaction was significant for the three traits, two inbred lines containing Z. diploperennis (ZD 551) and tropical (TZL TC 87) germplasm had negative and significant GCA effects for NESPP and HDS and positive GCA effects for grain yield under Striga infestation in the two countries. Correlation between NESPP and HDS was strong and significant (r = 0.87, P = 0.01). Our results highlight the importance of harnessing useful genes from wild relatives to improve resistance to S. hermonthica in adapted maize germplasm.
The barley mutant emr2 shows enhanced resistance against several fungal leaf pathogens
Authors: Jansen M. , Schaffrath U.
Published: February 2009

Homozygous mlo‐barley plants are resistant to barley powdery mildew but hypersusceptible to the rice blast fungus Magnaporthe oryzae. A mutational analysis was performed in the barley back‐cross line BCIngrid mlo5 which led to the identification of two mutants with enhanced capacity to resist infections by M. oryzae, referred to as enhanced M. oryzae resistance mutants emr1 and emr2. Here, we report on the characterization of emr2 mutant plants which not only show an almost complete reduction in disease severity after inoculation with M. oryzae but are also resistant to the necrotrophic fungi Drechslera teres and Rhynchosporium secalis. Histological analysis revealed that resistance to M. oryzae was based mainly on the formation of papillae at sites of attempted penetration into epidermal cells. There was no progression of fungal growth into the mesophyll. Additionally, because of the presence of the mlo‐allele, emr2‐plants retained resistance to powdery mildew. The emr2‐conditioned broad spectrum resistance was inherited as in a recessive manner. Monitoring of PR‐gene expression and enzymatic activity of peroxidases revealed a constitutively activated defence in emr2.
Molecular breeding for resistance toPhytophthora infestans(Mont.) de Bary in potato (Solanum tuberosumL.): a perspective of cisgenesis
Authors: Park T.‐H. , Vleeshouwers V. G. A. A. , Jacobsen E. , Van Der Vossen E. , Visser R. G. F.
Published: February 2009

Late blight caused byPhytophthora infestansis one of the most devastating diseases in potato cultivation and is mostly controlled by the application of chemicals. However, introduction of combinations of resistance (R) genes conferring broad‐spectrum resistance from wildSolanumspecies into cultivated potatoes is considered the most practical and promising approach to achieve durable resistance. This can be realized via classical breeding or genetic modification (GM). Because classical breeding is very time‐consuming and is often hampered by linkage drag, a GM approach seems logic in this heterozygous and vegetatively propagated crop. During the last decades, manyRgenes have been identified in several wildSolanumspecies. Some have been cloned and more will follow. When these genes are derived from species crossable with cultivated potato (so‐called cisgenes), application in resistance breeding using a GM approach is similar to an introgression breeding approach, in that the exploited genes are indigenous to the crop. Pending deregulation or derogation of cisgenesis, the use of cisgenicRgenes would be an ideal strategy to accomplish durable resistance in potato.
PREMATURE VENTRICULAR CONTRACTIONS ORIGINATING FROM THE RIGHT VENTRICULAR OUTFLOW TRACT: THREE‐DIMENSIONAL DISTRIBUTION OF THE TARGET SITES AND THEIR ELECTROCARDIOGRAPHIC CHARACTERISTICS
Authors: Yu Jian , Yang Bing , Chen Ming‐Long , Chen Hong‐Wu , Ju Wei‐Zhu , Shan Qi‐Jun , Zou Jian‐Gang , Chen Chun , Xu Dong‐Jie , Hou Xiao‐Feng , Cao Ke‐Jiang
Published: February 2009

The purpose of the present study was to explore the relationship between electrocardiogram (ECG) patterns of right ventricular outflow tract (RVOT) premature ventricular contractions and the three‐dimensional distribution of the target sites.

Thirty‐three consecutive patients were included in the study. The target sites were identified by non‐contact mapping and confirmed by successful ablation. The distribution of the target sites in the three‐dimensional reconstructed geometry of the RVOT was classified in three directions: (i) anterior (A)/posterior (P); (ii) free wall (F)/septal (Se); and (iii) superior (Su)/inferior (I). The ECG characteristics were then analysed according to the three‐dimensional distribution of the target sites.

The following indices were helpful to identify the position of the target site: (i) QRS duration (≥ 150 msec = F; < 150 msec = Se; P < 0.05); (ii) the R wave pattern in the inferior leads (RR′ or Rr′ = F; R = Se; P < 0.05); (iii) the R wave amplitude in the inferior leads (high = Se; low = F; P < 0.05); (iv) the initial r wave width in lead V1 (wide = F; narrow = Se; P < 0.05); (v) the QS wave amplitude in aVR and aVL (if aVR < aVL, A; if aVR ≥ aVL, P; P < 0.05); and (vi) the initial r wave amplitude in lead V1 and V2 (if V1 ≥ 0.15 mV and V2 ≥ 0.3 mV, Su; if V1 < 0.15 mV or V2 < 0.3 mV, I; P < 0.05).

In conclusoin, the ECG characteristics were associated with target site locations in all three directions.