bluuebunny/crossref_metadata_2025 huggingface dataset:
Content of ecstasy in the Netherlands: 1993–2008
Authors: Vogels Neeltje , Brunt Tibor M. , Rigter Sander , Van Dijk Peter , Vervaeke Hylke , Niesink Raymond J. M.
Published: October 2009

Aims  The present paper outlines the results of analyses carried out on the content of tablets sold as ecstasy, collected in the Netherlands by the Drugs Information Monitoring System (DIMS) from January 1993 to December 2008.

Methods  During a period of 16 years, the DIMS analysed the content of 33 006 tablets sold as ecstasy that were handed in by numerous individual (potential) substance users. The DIMS results were compared with the results from various seized tablets to determine whether the DIMS is a monitor of the ecstasy consumer market.

Results  The DIMS system appears to be a market monitor that gives an accurate reflection of what is actually available on the hidden Dutch ecstasy market. During 16 years of monitoring, the purity [tablets containing only 3,4‐methylenedioxymethamphetamine (MDMA)] was lowest around 1997. During this time‐period many tablets contained other substances in addition to or instead of MDMA [e.g. 3,4‐methylene‐dioxyamphetamine (MDA), 3,4‐methylene‐dioxyethylamphetamine (MDEA) and N‐methyl‐a‐(1,3‐benzodixol‐5‐yl)‐2‐butamine (MBDB), amphetamine and caffeine]. From 1998 to 2008, the number of high‐dose tablets (≥106 mg MDMA per tablet) gradually increased. The same holds true for the proportion of tablets that contained only MDMA, reaching the highest levels in 2000 and 2004. After 2004, the purity of ecstasy tablets decreased again, caused mainly by a growing proportion of tablets containing meta‐chlorophenylpiperazine (mCPP).

Conclusions  The DIMS results provide valuable qualitative information on the content of ecstasy tablets in the Netherlands, and its changes throughout the years. Moreover, the results were used for national and international risk assessments and important warning and prevention activities.
Extending drug ethno‐epidemiology using agent‐based modelling
Authors: Moore David , Dray Anne , Green Rachael , Hudson Susan L. , Jenkinson Rebecca , Siokou Christine , Perez Pascal , Bammer Gabriele , Maher Lisa , Dietze Paul
Published: October 2009

Aims  To show how the inclusion of agent‐based modelling improved the integration of ethno‐epidemiological data in a study of psychostimulant use and related harms among young Australians.

Methods  Agent‐based modelling, ethnographic fieldwork, in‐depth interviews and epidemiological surveys.

Setting  Melbourne, Perth and Sydney, Australia.

Participants  Club drug users in Melbourne, recreational drug users in Perth and street‐based injecting drug users in Sydney. Participants were aged 18–30 years and reported monthly or more frequent psychostimulant use.

Findings  Agent‐based modelling provided a specific focus for structured discussion about integrating ethnographic and epidemiological methods and data. The modelling process was underpinned by collective and incremental design principles, and produced ‘SimAmph’, a data‐driven model of social and environmental agents and the relationships between them. Using SimAmph, we were able to test the probable impact of ecstasy pill‐testing on the prevalence of harms—a potentially important tool for policy development. The study also navigated a range of challenges, including the need to manage epistemological differences, changes in the collective design process and modelling focus, the differences between injecting and non‐injecting samples and concerns over the dissemination of modelling outcomes.

Conclusions  Agent‐based modelling was used to integrate ethno‐epidemiological data on psychostimulant use, and to test the probable impact of a specific intervention on the prevalence of drug‐related harms. It also established a framework for collaboration between research disciplines that emphasizes the synthesis of diverse data types in order to generate new knowledge relevant to the reduction of drug‐related harms.
Influence of smoking cues in movies on craving among smokers
Authors: Lochbuehler Kirsten , Engels Rutger C. M. E. , Scholte Ron H. J.
Published: October 2009

Aims  Research has shown that smoking‐related cues are important triggers for craving. The objective of the present study was to test whether smoking cues in movies also function as triggers to evoke craving. To accomplish this, we conducted a pilot study in which we examined smokers' reactivity to smoking cues from a particular movie in a common cue–reactivity paradigm using pictures. In the main study, we tested whether smokers who are confronted with smoking characters in a movie segment have a greater desire to smoke than smokers confronted with non‐smoking characters.

Design  Using an experimental design, participants were assigned randomly to one of two movie conditions (smoking versus non‐smoking characters).

Setting  In a laboratory, that reflected a naturalistic setting, participants watched a 41‐minute movie segment.

Participants  A total of 65 young adults who smoked on a daily basis participated in the experiment.

Measurements  Craving was assessed before and after watching the movie.

Findings  The pilot study revealed that pictures of smoking characters had strong effects on craving. However, when smokers actually watched a movie segment, no differences in craving were found between those who watched smoking characters and those who watched non‐smoking characters. This finding was not affected by baseline craving, the time of the last cigarette smoked and daily smoking habits.

Conclusions  No effect of smoking cues in movies on craving was found, in contrast with research supporting the cue‐craving link. Thus, if replicated, this might indicate that smoking cues in such contexts do not affect smokers' desire to smoke as expected.
An audit of intensive care unit recyclable waste
Authors: McGain F. , Story D. , Hendel S.
Published: October 2009

There is little known about recyclable intensive care unit waste. We tested the hypotheses that the intensive care unit produces a small proportion (< 10%) of hospital waste, that much waste (> 30%) is recyclable and that there is little (< 10%) cross‐contamination of non‐infectious with infectious waste. For seven consecutive days in an Australian 10‐bedded intensive care unit, we prospectively sorted all waste. The total intensive care unit waste for the week was 540 kg, representing 5% of hospital waste. Of the 401 kg of intensive care unit general waste, recyclables were 230 kg (57%; 95% CI 53–61%), mainly plastics, cardboard and paper. There were 0.4 kg of infectious cross‐contamination in the 401 kg of general waste. Intensive care unit waste was a small proportion of all hospital waste. However, there was minimal infectious waste cross‐contamination and almost 60% of intensive care unit general waste could be recycled with appropriate safeguards, education and training.
Failure of three novel regimens to improve outcome for patients with relapsed or refractory acute myeloid leukaemia: a report from the Eastern Cooperative Oncology Group
Authors: Litzow Mark R. , Othus Megan , Cripe Larry D. , Gore Steven D. , Lazarus Hillard M. , Lee Sandra J. , Bennett John M. , Paietta Elisabeth M. , Dewald Gordon W. , Rowe Jacob M. , Tallman Martin S. ,   for the Eastern Cooperative Oncology Group Leukemia Committee
Published: October 2009

The treatment of relapsed acute myeloid leukaemia (AML) remains unsatisfactory. We conducted a phase II randomized trial where patients received intermediate‐dose cytarabine for 4 d followed by gemtuzumab ozogamicin on day 5 (Arm A), or combined with liposomal daunorubicin for 3 d (Arm B), or cytarabine given for 5 d combined with cyclophosphamide for 3 d and topotecan by continuous infusion for 5 d (Arm C). Eligible patients had primary refractory AML, a first relapse after a remission of <1 year, or a second or greater relapse. The primary objective of this trial was attainment of a conventional complete remission (CR) or a CR without platelet recovery (CRp) in at least 40% of patients. The CR/CRp rates for the 82 eligible patients were 3/26 (12%) in Arm A, 2/29 (7%) in Arm B, and 1/27 (4%) in Arm C. No patients who had relapsed within 6 months of initial CR or who had suffered multiple relapses responded. More than 95% of patients subsequently died of AML. No unexpected toxicities were encountered. We conclude that none of these three regimens were effective enough in the treatment of high‐risk relapsed or refractory AML to warrant further study. This trial was registered at http://www.clinicaltrials.gov as #NCT00005962.
Combination of KIT gene silencing and tocopherol succinate may offer improved therapeutic approaches for human mastocytosis
Authors: Ruano Irene , Gargini Ricardo , Izquierdo Marta
Published: October 2009

Gain‐of‐function mutations of kit tyrosine kinase receptor are associated with mastocytosis. Two subclones of the HMC1 mast leukaemia cell line were used; both express an identical KIT allele‐specific regulatory type mutation (V560G), but differ in that one also expresses an enzymatic site type mutation (D816V) that confers on them resistance to imatinib mesylate tyrosine kinase inhibitor. In both cell lines, proliferation was suppressed and apoptosis induced by the combination of KIT gene silencing and α‐tocopherol succinate (α‐TOS), a derivate of α‐tocopherol, also known as vitamin E. Furthermore, HMC1 cells with decreased kit levels by KIT silencing, failed to form tumours when xenotransplanted into immunocompromised mice and the animals were treated systemically with α‐TOS. Targeting kit in the presence of α‐TOS represents a new approach against proliferation of human mast leukaemia cell lines.
Hearing the child
Authors: Archard David , Skivenes Marit
Published: October 2009

Given that in our view the child has a fundamental right to be heard in all collective deliberative processes determining his or her future, we set out, firstly, what is required of such processes to respect this right – namely that the child's authentic voice is heard and makes a difference – and, secondly, the distance between this ideal and practice exemplified in the work of child welfare and child protection workers in Norway and the UK, chiefly in their display of an instrumental attitude to children's views.
Relationships matter: the problems and prospects for social workers’ relationships with young children in care
Authors: Winter Karen
Published: October 2009

One of the key lessons learnt in the UK from the Laming Inquiry into the death of Victoria Climbié was the importance of social workers developing consistent and long‐term relationships with young children in whose lives they are involved. This issue is now informing policy developments, including the proposed Social Work Practices which, based on a similar model to General Practitioner practices, aim to provide a lead professional to act as a parental figure and an advocate for every child in care. This paper begins by confirming the importance of developing relationships between social workers and young children, but questions the ability of the new policy developments to facilitate these. Drawing upon data from research involving interviews with social workers, the paper outlines the factors which hinder social workers’ relationships with young children and argues that while the new proposals address some of the more surface structural and organizational factors, they do not address the deeper factors regarding attitudes, values and emotional competence which are crucial if social workers are to successfully build relationships with young children in care.
Communication with parents in child welfare: skills, language and interaction
Authors: Hall Christopher , Slembrouck Stef
Published: October 2009

This paper reviews the literature on communication between professionals and parents/carers in health and social care. It notes that there is considerable research on doctor–patient communication and counselling, but much less on professionals who visit families in their homes. While there is a large literature on good practice and communication skills, there is little research on the actual encounters themselves. What counts as ‘communication’ in such publications is explored. The paper suggests that there is much to be gained by considering research in discourse analysis and sociolinguistics on professional–client encounters. Examples of some of the key concepts are discussed and an engagement with this literature is recommended.
Anxiety and child protection – implications for practitioner–parent relations
Authors: Waterhouse Lorraine , McGhee Janice
Published: October 2009

Social work practitioners face powerful existential threats in child protection. Core aspects of anxiety and their significance for practitioner–parent relations are identified. Social and psychoanalytic theories are utilized to suggest that the production of anxiety in child protection stems from multiple sources and that insufficient attention has been paid to the social context of poverty and disadvantage. Menzies' core analytic categories of primary and secondary anxiety are applied and extended to take account of specific dimensions of the social and organizational context of child protection practice for social work. Processing the anxiety is central to practitioner–parent relations that lie at the heart of the protection of children.
Alkaloids may not be responsible for endophyte‐associated reductions in tall fescue decomposition rates
Authors: Siegrist Jacob A. , McCulley Rebecca L. , Bush Lowell P. , Phillips Tim D.
Published: October 2009

1.  Fungal endophyte – grass symbioses can have dramatic ecological effects, altering individual plant physiology, plant and animal community structure and function, and ecosystem processes such as litter decomposition and nutrient cycling.

2.
 Within the tall fescue (Schedonorus arundinaceus) – fungal endophyte (Neotyphodium coenophialum) symbiosis, fungal produced alkaloids are often invoked as the putative mechanism driving these ecological responses. Yet few measurements of alkaloids exist in the ecological literature. In this study, we quantified alkaloid levels in live, standing dead and decomposing endophyte‐infected (E+) and –free (E−) plant material and simultaneously evaluated the direct and indirect effects of endophyte presence on tall fescue decomposition.

3.
 Loline and ergot alkaloid levels were consistently high in live E+ (common toxic strain of N. coenophialum) tall fescue biomass throughout the sampling period (May–November 2007), whereas, E− live and standing dead material had non‐detectable alkaloid concentrations. Standing dead E+ biomass had significantly reduced alkaloid levels (6–19x lower than the levels measured in the corresponding live E+ biomass) that were equivalent to E− live and dead for loline but were still somewhat higher than E− material for ergots.

4.  In an effort to test the role of alkaloids in directly inhibiting decomposition, as has been suggested by previous studies, we conducted a litter bag experiment using green, alkaloid‐laden E+ and alkaloid‐free E− tall fescue plant material. We incubated E+ and E− litter bags in both E+ and E− tall fescue stands for 170 days, and measured mass loss, carbon and nitrogen content, and ergot and loline alkaloid concentrations over the incubation period.

5.  Consistent with previous reports, both direct and indirect effects of endophyte presence on litter decomposition were observed: endophyte presence in the litter and surrounding microenvironment significantly reduced decomposition rates. Surprisingly, despite large differences in alkaloid content between E+ and E− litter from Day 0–Day 21 of the incubation, direct effects of the endophyte on litter decomposition, while significant, were relatively small (differences in mass loss between E+ and E− litter were never >3%). Alkaloids were gone from E+ material by day 56.

6.  We propose that results from this study indicating alkaloids are largely absent in standing dead material (the typical input to the decomposition process), and that despite being present in our litter bag experiment, failed to produce large differences in mass loss between E+ and E− material questions the supposition that fungal produced alkaloids directly inhibit decomposition. Additional studies exploring the mechanisms behind the direct and indirect effects of endophytes on this ecosystem process are needed.
Influence of aquatic training on the motor performance of patients with haemophilic arthropathy
Authors: VALLEJO L. , PARDO A. , GOMIS M. , GALLACH J. E. , PÉREZ S. , QUEROL F.
Published: October 2009

Summary.  Thirteen patients with haemophilia A took part in this study voluntarily. They underwent an aquatic training programme over a 9‐week period (27 sessions; three sessions per week; 1 h per session). Their motor performance was assessed by the following cardio‐respiratory and mechanical variables before and after the training programme: oxygen uptake (VO2, mL min−1), relative oxygen uptake (rel VO2, mL min−1·kg−1), carbon dioxide (CO2, mL min−1), respiratory quotient (R), heart rate (bpm) and the distance covered in 12 min (the Cooper test, m). Nine patients successfully completed the intervention and measurement protocols without bleeding or other adverse events. After the proposed training programme, significant differences between the pre‐test and post‐test were observed. Patients’ aerobic capacity increased considerably, and their oxygen uptake improved by 51.51% (P < 0.05), while their relative oxygen uptake went up by 37.73% (P < 0.05). Their mechanical capacity also increased considerably (14.68%, P < 0.01). Our results suggest that 27 specially designed aquatic training sessions for our patients with haemophilia A had a positive effect on their motor performance and considerably improved their aerobic and mechanical capacity without causing adverse effects.
Effects of competition on great and blue tit reproduction: intensity and importance in relation to habitat quality
Authors: Dhondt André A.
Published: October 2009

1. In studies on the effect of competition in plant communities two terms are used to describe its effects: the absolute reduction in growth of an individual as a consequence of the presence of another one is called intensity, while the relative impact of competition on an individual as a proportion of the impact of the whole environment is called importance. One school of thought is that the role of competition remains constant across productivity gradients, while the other is that it decreases with increasing severity. J.B. Grace (1991. A clarification of the debate between grime and tilman. Functional Ecology, 5, 583–587.) suggested that the apparent contradiction might be solved if we acknowledge that the two schools are discussing different aspects of competition: the intensity of competition might remain constant while its importance declines with increasing severity.

2. There are no studies that compare intensity and importance of competition in bird populations between areas that differ in quality or productivity and hence it is not possible to make predictions how intensity or importance of competition would vary between them.

3. I compared variation in intensity and importance of competition of three demographic variables between five plots that differ strongly in quality for great Parus major L. and blue tit Cyanistes caeruleus (L.).

4. Both intensity and importance of competition are larger in great than in blue tit populations meaning that the effect of competition on demographic variables is stronger in great than in blue tits and that the contribution of competition to variation in these variables is relatively higher in great than in blue tits.

5. Intensity of competition is higher in low quality than in high quality plots for both species, a result not expected from studies in plant communities.

6. Importance of competition varies strongly between plots. It is larger in oak‐dominated plots than in mixed deciduous plots.

7. In birds breeding density increases with habitat quality but is limited by territorial behaviour. As a result competition for food is reduced in high quality habitats resulting in a reduction of competition intensity in high quality sites in which birds breed at high densities.

8. It can be predicted that in studies of territorial species density dependent effects on reproduction are more likely to be detected in low quality sites explaining in part differences in results between studies.
Optimal combinations of ultrasound‐based and serum markers of disease severity in patients with chronic hepatitis C
Authors: Cobbold J. F. L. , Crossey M. M. E. , Colman P. , Goldin R. D. , Murphy P. S. , Patel N. , Fitzpatrick J. , Vennart W. , Thomas H. C. , Cox I. J. , Taylor‐Robinson S. D.
Published: October 2009

Summary. Combinations of noninvasive markers may improve discrimination of chronic liver disease severity. The aims of this study were to compare four validated serum and ultrasound‐based markers of hepatic disease severity head‐to‐head with liver biopsy and to assess optimal combinations with consideration of cost. A total of 67 patients with biopsy‐proven chronic hepatitis C underwent all four techniques on the same visit [aspartate aminotransferase (AST) to platelet ratio index (APRI); Enhanced Liver Fibrosis (ELF) panel; transient elastography (TE) and ultrasound microbubble hepatic transit times (HTT)]. Markers were combined according to increasing financial cost and ordinal regression used to determine contributions. APRI, ELF, TE and HTT predicted cirrhosis with diagnostic accuracy of 86%, 91%, 90% and 83% respectively. ELF and TE were the most reliable tests with an intra‐class correlation of 0.94 each. Either ELF or TE significantly enhanced the prediction of fibrosis stage when combined with APRI, but when combined together, did not improve the model further. Addition of third or fourth markers did not significantly improve prediction of fibrosis. Combination of APRI with either ELF or TE effectively predicts fibrosis stage, but combinations of three or more tests lead to redundancy of information and increased cost.
Multiyear multiple paternity and mate fidelity in the American alligator,Alligator mississippiensis
Authors: LANCE S. L. , TUBERVILLE T. D. , DUECK L. , HOLZ‐SCHIETINGER C. , TROSCLAIR P. L. , ELSEY R. M. , GLENN T. C.
Published: October 2009

We examined multiple paternity during eight breeding events within a 10‐year period (1995–2005) for a total of 114 wild American alligator nests in Rockefeller Wildlife Refuge in south‐west Louisiana. Our goals included examining (i) within population variation in multiple paternity among years, (ii) variation in multiple paternity in individual females and (iii) the potential for mate fidelity. To accomplish this, in the current study, eggs were sampled from 92 nests over 6 years and analysed along with 22 nests from a previous 2‐year study. Genotypes at five microsatellite loci were generated for 1802 alligator hatchlings. Multiple paternity was found in 51% of clutches and paternal contributions to these clutches were highly skewed. Rates of multiple paternity varied widely among years and were consistently higher in the current study than previously reported for the same population. Larger females have larger clutches, but are not more likely to have multiply sired nests. However, small females are unlikely to have clutches with more than two sires. For 10 females, nests from multiple years were examined. Seven (70%) of these females exhibited long‐term mate fidelity, with one female mating with the same male in 1997, 2002 and 2005. Five females exhibiting partial mate fidelity (71%) had at least one multiple paternity nest and thus mated with the same male, but not exclusively. These patterns of mate fidelity suggest a potential role for mate choice in alligators.
Nuclear and mitochondrial DNA reveals isolation of imperilled grey nurse shark populations (Carcharias taurus)
Authors: AHONEN H. , HARCOURT R. G. , STOW A. J.
Published: October 2009

Loss of sharks and other upper‐trophic marine predators has sparked worldwide concern for the stability of ocean ecosystems. The grey nurse (ragged‐tooth or sand tiger) shark (Carcharias taurus) is Vulnerable on a global scale, Critically Endangered in Australia and presumed extinct in parts of its historical range. We used 193 muscle and fin samples collected from six extant populations to assess global mtDNA and microsatellite diversity and the degree of global population genetic structure. Control region mtDNA diversity was low in every population, and two populations (eastern Australia and Japan) contained only a single mtDNA haplotype. Genetic signatures of recent losses of genetic variation were not yet apparent at microsatellite loci, indicating that this low mtDNA variation is not a result of anthropogenic population declines. Population differentiation was substantial between each population pair except Brazil and South Africa, FST values ranged from 0.050 to 0.699 and 0.100 to 1.00 for microsatellite and mitochondrial data respectively. Bayesian analysis clearly partitioned individuals into five of the populations from which they were sampled. Our data imply a low frequency of immigrant exchange among each of these regions and we suggest that each be recognized as a distinct evolutionary significant unit. In contrast to pelagic species such as whale shark and white shark that may cross ocean basins and where cooperative international efforts are necessary for conservation, grey nurse shark, like many coastal species, need to be managed regionally.
Specific detection of Phytophthora cactorum in diseased strawberry plants using nested polymerase chain reaction
Authors: Bhat R. G. , Browne G. T.
Published: October 2009

Validated protocols for DNA purification and PCR amplification are reported for detection of Phytophthora cactorum in diseased strawberry plants. To remove PCR inhibitors, necrotic strawberry tissues were soaked in 5% alconox solution for >12 h before DNA extraction, and the extracted genomic DNA was embedded in an agarose gel chamber and subjected to electrophoresis. The purified DNA was amplified reliably by PCR. Nested PCR was used to detect a portion of the rRNA gene of P. cactorum in samples. In the first round of PCR, primers ITS1 and ITS4 amplified fragments of varying sizes from total genomic DNA from diseased strawberry plants. In the second round of PCR, a 1:25 dilution of the first‐round PCR products was used as template with two P. cactorum‐specific primer pairs (BPhycacL87FRG and BPhycacR87RRG, which amplified a 340‐bp fragment and a 480‐bp fragment from the rRNA gene; and BPhycacL89FRG and BPhycacR176RRG, which amplified a 431‐bp fragment). Validation tests using culture‐based isolations as a standard for comparison indicated that the DNA purification and PCR primers and amplification protocols were reliable and specifically amplified a portion of the rRNA gene of P. cactorum from necrotic root, crown and petiole tissues of strawberry naturally infected by the pathogen.
Occurrence of Corynespora cassiicola isolates resistant to boscalid on cucumber in Ibaraki Prefecture, Japan
Authors: Miyamoto T. , Ishii H. , Seko T. , Kobori S. , Tomita Y.
Published: October 2009

A total of 651 isolates of cucumber corynespora leaf spot fungus (Corynespora cassiicola) collected from cucumber in Japan, either with (438 isolates) or without (213 isolates) a prior history of boscalid use, were tested for their sensitivity to boscalid by using a mycelial growth inhibition method on YBA agar medium. Additionally, seven isolates of C. cassiicola obtained from tomato, soybean, eggplant (aubergine) and cowpea in different locations in Japan were tested before boscalid registration. Minimum inhibitory concentration (MIC) and 50% effective concentration (EC50) values for 220 isolates from crops without a prior history of boscalid use ranged from 0·5 to 7·5 μg mL−1 and from 0·04 to 0·59 μg mL−1, respectively. Two hundred and fourteen out of 438 isolates collected from ten cucumber greenhouses in Ibaraki Prefecture, Japan, which received boscalid spray applications showed boscalid resistance, with MIC values higher than 30 μg mL−1. Moreover, resistant isolates were divided into two groups: a moderately resistant (MR) group consisting of 189 isolates with EC50 values ranging from 1·1 to 6·3 μg mL−1, and a very highly resistant (VHR) group consisting of 25 isolates with EC50 values higher than 24·8 μg mL−1. MR isolates were detected from all ten greenhouses, but VHR isolates were detected from only three. As a result of fungus inoculation tests which used potted cucumber plants, control failures of boscalid were observed against resistant isolates. Efficacy of boscalid was remarkably low against VHR isolates in particular. This is the first known report on boscalid resistance in Japan.
Detection of Ralstonia solanacearum (biovar 2A) in stems of symptomless plants before harvest of the potato crop using post‐enrichment DAS‐ELISA
Authors: Priou S. , Gutarra L. , Aley P. , De Mendiburu F. , Llique R.
Published: October 2009

The detection of Ralstonia solanacearum (biovar 2A) in stems of symptomless plants before harvest of the potato crop, instead of tubers, would not only save highly valued planting material but would be less time‐consuming and would also enhance farmers’ market decisions. Although pathogen detection in stems has been proven efficient for ring rot, this has never been investigated for bacterial wilt (BW). Therefore the possibility of detecting BW latent infection in stem pieces about three weeks before harvest was assessed in 57 fields of the Andean highlands of Peru. Two sensitive, specific and user‐friendly serological methods were used to detect the pathogen in latently infected tubers and stems: double‐antibody sandwich (DAS)‐ELISA and indirect ELISA on nitrocellulose membrane (NCM) after enrichment of the plant extracts in a semi‐specific broth. Optimum sample sizes of stems and tubers were evaluated for 37 potato crops showing between 0 and 0·1% BW incidence using a binomial distribution model to calculate the detection probabilities. Although results of detection using the two serological techniques had 100% concordance, detection probabilities were higher using DAS‐ELISA, whatever the plant part tested. BW detection probabilities were higher for tubers than for stems; a 99% detection probability was obtained by analysing 400 stems sections or 250 tubers using DAS‐ELISA. Detection of BW infection in symptomless plants 20 days before harvest using post‐enrichment DAS‐ELISA is a reliable and user‐friendly technique that can easily be used by national plant protection services and seed programmes in developing countries.
Detection and molecular characterization of Italian Grapevine rupestris stem pitting‐associated virus isolates
Authors: Terlizzi F. , Ratti C. , Filippini G. , Pisi A. , Credi R.
Published: October 2009

The efficiencies of two published primer pairs (RSP13‐RSP14 and RSP2‐RSP21) were compared in reverse transcription‐polymerase chain reaction (RT‐PCR) to improve the diagnosis of Grapevine rupestris stem pitting‐associated virus (GRSPaV), which has consistently been found associated with rupestris stem pitting (RSP), one of the most common infectious diseases of grapevine. Fifty‐six grapevine accessions from different locations in Northern Italy were tested. Of these, 48 were positive with primers RSP13‐RSP14 and 37 with primers RSP2‐RSP21, suggesting that primer pair RSP13‐RSP14 has a broader detection spectrum than primers RSP2‐RSP21. The coat protein (CP) gene was also amplified by RT‐PCR and the amplicons analysed by single‐strand conformation polymorphism (SSCP) and sequencing. The results support the existence of wide variability within this virus, independent of cultivar or geographic origin. Sequence analysis of the 17 Italian GRSPaV isolates examined showed a molecular identity ranging from 78·8 to 99·7%, within which, four groups of sequence variants were recognized. Flexuous rod‐shaped particles, approximately 800 nm in length, were detected in plant extracts from infected grapevines and were positively decorated with the specific antiserum As7‐276 in immunosorbent electron microscopy (ISEM). This is the first report of GRSPaV particles in grapevines observed by transmission electron microscopy (TEM) in Italy.
Rapid emergence of hybrids between the two subspecies ofOphiostoma novo‐ulmiwith a high level of pathogenic fitness
Authors: Brasier C. M. , Kirk S. A.
Published: October 2009

During the 1970s Europe was invaded by two subspecies of the Dutch elm disease pathogenOphiostoma novo‐ulmi: subsp. americanafrom the west and subsp.novo‐ulmifrom the east. As a result their geographic ranges began to overlap in several areas. Only a weak prezygotic barrier to hybridization exists between the subspecies and in 1980 two hybrids were detected in the Netherlands. A subset of 107O. novo‐ulmiisolates collected in a subspecies overlap zone in Limburg, Netherlands in 1983 was characterized for three phenotypic markers and seven RAPD PCR markers. By phenotype, 33% were shown to be hybrid whereas by RAPD markers 69% were shown to be hybrid. Some isolates shown to be hybrid by phenotype were not revealed to be hybrid by PCR and vice versa. Combining the phenotype and RAPD data the estimated hybrid frequency was ∼78%. The mean growth rate of Limburg hybrid isolates was significantly faster than that of the Limburg subsp.novo‐ulmiisolates but not significantly different from Limburg subsp.americanaisolates. The Limburg hybrid isolates were just as pathogenic as the parent subspecies on both clonalUlmus proceraand onU. × Commelin. A subset of 100 isolates collected in another subspecies overlap zone at Orvieto, Italy in 1986 was also assessed with RAPD markers and∼72% were shown to be hybrids. When 20 isolates of a ‘pure’ subsp.novo‐ulmipopulation in the Baltic Ports area of Poland collected in 1980 were assessed by RAPD markers three isolates exhibited early introgression of subsp.americanaDNA. This study therefore demonstrates very rapid emergence ofO. novo‐ulmisubspecies hybrids and introgressants in Europe in the early 1980s. In terms of two major fitness characters, growth rate and pathogenicity, these early hybrids were as fit as their parent subspecies. It is likely that complex hybrid swarms are now expanding across the continent.
PVYNTN‐NW, a novel recombinant strain of Potato virus Y predominating in potato fields in Syria
Authors: Chikh Ali M. , Maoka T. , Natsuaki T. , Natsuaki K. T.
Published: October 2009

Detailed characterization of a number of isolates of PVYSYR, a novel recombinant strain of Potato virus Y (PVY) from Syria, was conducted to elucidate their origin, assess their significance and achieve a final classification of PVYSYR. Recombination analysis grouped isolates of PVYSYR into three recombination patterns, SYR‐I, SYR‐II and SYR‐III, which varied in the first 700 nucleotides of their genomes, with the second recombination pattern, SYR‐II, the most frequent. PVYSYR isolates shared highest genomic identity and close phylogenetic relationships with PVYNTN and PVYNW isolates from Syria, suggesting a common origin and local emergence of these isolates in Syria. All PVYSYR isolates (total of 20) induced tobacco veinal necrosis, but reacted to a PVYO monoclonal antibody, typical characteristics of the previously reported PVYNW (or PVYN:O). In potato, however, four isolates tested (one of SYR‐I and three of SYR‐II) induced potato tuber necrotic ringspot disease (PTNRD), which is the characteristic phenotype of PVYNTN. Given the shared properties of SYR‐I and SYR‐II isolates with PVYNTN and PVYNW, it was decided that they represent a new recombinant strain of the PVYN strain group, with the proposed name PVYNTN‐NW. The classification of SYR‐III will be possible only after testing the phenotype in potato. The high prevalence of PVYNTN‐NW in potatoes and weeds, as well as its ability to induce PTNRD, demonstrates its importance and the necessity for its control.
Matrix metalloproteinases‐7, ‐8, ‐9 and TIMP‐1 in the follow‐up of diisocyanate‐induced asthma
Authors: Piirilä P. , Lauhio A. , Majuri M.‐L. , Meuronen A. , Myllärniemi M. , Tervahartiala T. , Vuorinen K. , Laitinen A. , Alenius H. , Kinnula V. L. , Sorsa T.
Published: October 2009

Background:  Diisocyanate‐induced asthma (DIA) is known to be associated with poor prognosis. We wished to clarify if matrix metalloproteinases (MMP)‐7, ‐8 or ‐9 or tissue inhibitor of matrix metalloproteinases (TIMP‐1) are associated with the functional or inflammatory outcome in DIA patients.

Methods:  This is a longitudinal study where 17 patients with DIA diagnosed by a specific challenge test to diisocyanates were monitored. Exposure to diisocyanates was terminated seven (mean) months before the challenge test. The studies included spirometry, histamine challenge test and bronchoscopy. MMP‐7, MMP‐8, TIMP‐1 [Enzyme‐linked immunosorbent assay (ELISA)‐ and immunofluorometric assay‐methods], MMP‐9 (ELISA and zymography), interferon‐gamma, tumour necrosis factor‐alpha, interleukin‐6, ‐8, ‐15, ‐17, CXCL‐5/ENA‐78, monocyte chemoattractant protein‐1 and macrophage inhibitory factor (MIF) (ELISA) were assayed from bronchoalveolar lavage (BAL) fluid. Inhaled steroid therapy was initiated after the examinations, which were repeated at 6 months and at 3 years during the treatment. The results were compared with those of 15 healthy controls.

Results:  Inhaled steroid medication increased BAL levels of MMP‐9 and MMP‐9/TIMP‐1 and decreased MMP‐7 and MMP‐7/TIMP‐1. The increase in MMP‐9 levels was associated with a decline in the TH‐2 type inflammation.

Conclusions:  Our data suggest that reduced TH‐2 type inflammation in DIA after inhaled steroid medication is reflected as elevated MMP‐9 and MMP‐9/TIMP‐1 levels in BAL. MIF may be the inducer of MMP‐9. This might point to some protective role for MMP‐9 in DIA.
Temperature modulated histamine‐itch in lesional and nonlesional skin in atopic eczema – a combined psychophysical and neuroimaging study
Authors: Pfab F. , Valet M. , Sprenger T. , Huss‐Marp J. , Athanasiadis G. I. , Baurecht H. J. , Konstantinow A. , Zimmer C. , Behrendt H. , Ring J. , Tölle T. R. , Darsow U.
Published: October 2009

Background:  Itch is the major symptom of many allergic diseases; yet it is still difficult to measure objectively. The aim of this study was to use an evaluated itch stimulus model in lesional (LS) and nonlesional (NLS) atopic eczema (AE) skin and to characterize cerebral responses using functional magnetic resonance imaging (fMRI).

Methods:  Thermal modulation was performed on a histamine stimulus in randomized order on LS or NLS in rapid alternating order from 32°C (warm) to 25°C (cold). Subjective itch ratings were recorded. Additionally, fMRI measurements were used to analyze the cerebral processing (n = 13). Healthy skin (HS) of age‐matched volunteers served as control (n = 9).

Results:  Mean VAS itch intensity was significantly (P < 0.0001) higher during the relative cold [55.2 ± 8.3% (LS); 48.6 ± 8.2% (NLS)] compared to the relative warm blocks [36.0 ± 7.3% (LS); 33.7 ± 7.6% (NLS)]. Compared to HS, the itch response was delayed in LS and NLS. Itch intensity was perceived highest in LS, followed by NLS and HS. For NLS, fMRI revealed at the beginning of the itch provocation a cerebral deactivation pattern in itch processing structures (thalamus, prefrontal, cingulate, insular, somatosensory and motor cortex). During the course of stimulation, the cerebral deactivation was reduced with time and instead an activation of the basal ganglia occurred. In contrast LS showed an activation instead of deactivation pattern already at the beginning of the stimulation in the above mentioned structures.

Conclusions:  Moderate short‐term temperature modulation led to a reproducible, significant enhancement of histamine‐induced itch with the strongest effect in LS. The differences in itch perception and itch kinetics between healthy volunteers and NLS in patients point towards an ongoing central inhibitory activity patients with AE, especially at the beginning of the itch provocation.
The allergen specificity of the late asthmatic reaction
Authors: Hatzivlassiou M. , Grainge C. , Kehagia V. , Lau L. , Howarth P. H.
Published: October 2009

To cite this article: Hatzivlassiou M, Grainge C, Kehagia V, Lau L, Howarth PH. The allergen specificity of the late asthmatic reaction. Allergy 2010; 65: 355–358.

Background:  Allergen inhalation challenge in asthma may induce both early (EAR) and late (LAR) asthmatic reactions. The EAR is IgE and mast cell dependent. The mechanism of the LAR is less well defined and we have hypothesized may be allergen dependent. The aim of this study was to investigate the allergen specificity of the LAR to allergen inhalation in asthma.

Methods:  In a randomized, double‐blind, crossover design six asthmatic volunteers with dual sensitization to house dust mite (HDM) allergen and grass pollen (GP) allergen underwent inhalation allergen challenge with these separate allergens on two occasions separated by 14 days. Lung function changes were followed for 8‐h postchallenge. Bronchial reactivity (histamine PC20) and airway inflammation, assessed by induced sputum differential cell count, were measured 24‐h pre and postallergen challenge. The allergen inhalation challenges were matched to achieve the same magnitude of EAR.

Results:  Despite comparable group mean EAR percent falls in FEV1 (25.8% following GP and 28.0% following HDM (P = 0.917), the LAR was statistically greater on the HDM challenge day (13.0%vs 22.8% [P = 0.046]) and was associated with a significant airway eosinophil recruitment (mean (SD) of 5.4 (4.8)% to 22.1 (18.2)% (P = 0.028) that was not evident on the GP allergen challenge day.

Conclusions:  These findings identify the allergen specificity of the LAR and indicate that factors independent of IgE contribute to the LAR. Such findings have relevance both to the understanding of the allergen‐induced airway responses in asthma and the need for homogeneity in inhaled‐allergen challenge studies in asthma.
Steroid sparing effects of intranasal corticosteroids in asthma and allergic rhinitis
Authors: Nair A. , Vaidyanathan S. , Clearie K. , Williamson P. , Meldrum K. , Lipworth B. J.
Published: October 2009

To cite this article: Nair A, Vaidyanathan S, Clearie K, Williamson P, Meldrum K, Lipworth BJ. Steroid sparing effects of intranasal corticosteroids in asthma and allergic rhinitis. Allergy 2010; 65: 359–367.

Background:  Treating allergic rhinitis may have a downstream anti‐inflammatory effect on the lower airways. We conducted a dose ranging study in asthma and persistent allergic rhinitis to evaluate if intranasal corticosteroids exhibit a sparing effect on the dose of inhaled corticosteroid.

Methods:  Twenty five participants were randomized to receive two weeks of 100 μg/day (Low dose) or 500 μg/day (High dose) of inhaled fluticasone propionate both with intranasal placebo; or inhaled fluticasone 100 μg/day with intranasal fluticasone 200 μg/day (Combined) in a double‐blind cross‐over fashion.

Results:  Low dose fluticasone produced a shift of 1.20 doubling‐dilutions (95% CI, 0.63, 1.77); Combined fluticasone, 1.79 doubling‐dilutions (95% CI, 0.77, 2.80) and high dose fluticasone, 2.01 doubling‐dilutions (95% CI, 1.42, 2.61) in methacholine PC20 from respective baselines. There was a significant difference between high and low doses: 0.82 doubling dilutions (95%CI, 0.12, 1.50) but not between combined and low dose 0.58 doubling dilutions (95% CI, –0.78, 1.95). Combined treatment alone produced improvements in peak nasal inspiratory flow (P < 0.001), rhinitis quality of life (P = 0.004) and nasal NO (P = 0.01); reduced blood eosinophil count (P = 0.03), and serum eosinophil cationic protein (P = 0.02). All treatments significantly improved tidal NO, FEV1 and asthma quality of life.

Conclusions:  High‐dose fluticasone was superior to low dose fluticasone for methacholine PC20, demonstrating room for further improvement. Combined treatment was not significantly different from low dose fluticasone and we could not demonstrate a steroid sparing effect on methacholine PC20. Combined treatment alone produced improvements in upper airway outcomes and suppressed systemic inflammation but not adrenal function.
Passive smoking is a major determinant of exhaled nitric oxide levels in allergic asthmatic children
Authors: Laoudi Y. , Nikasinovic L. , Sahraoui F. , Grimfeld A. , Momas I. , Just J.
Published: October 2009

To cite this article: Laoudi Y, Nikasinovic L, Sahraoui F, Grimfeld A, Momas I, Just J. Passive smoking is a major determinant of exhaled nitric oxide levels in allergic asthmatic children. Allergy 2010; 65: 491–497.

Background:  Fraction of exhaled nitric oxide (FeNO) is considered, by some authors, to be a treatment follow‐up parameter in allergic asthmatics. However, factors such as active smoking can influence NO production and must be taken into account in the interpretation of FeNO values. In children, the evidence in favour of an impact of passive smoking (PS) on FeNO values is controversial. The aim of this study was to evaluate the impact of chronic PS on FeNO in allergic asthmatic children.

Methods:  Seventy nontreated allergic asthmatic children over 5 years of age, exposed and unexposed to PS, underwent measurement of FeNO, spirometry, and allergic tests (skin prick tests, total and specific serum IgE, and blood eosinophilia). Children were considered to be exposed to PS when at least 1 cigarette per day was declared to be smoked at home.

Results:  Geometric mean FeNO value in 22 children exposed to PS was 26.3 ± 1.5 ppb vs 56.3 ± 1.7 ppb in 48 children unexposed (P < 0.001). After adjustment for age, blood eosinophilia, allergic sensitizations, total IgE, dust mite sensitization and asthma severity, multivariate analysis showed that PS exposure was negatively associated with FeNO values (P = 0.0001) and was the primary determinant of FeNO variations.

Conclusion:  Passive smoking lowers FeNO, and might be a major determinant of FeNO levels in nontreated allergic asthmatic children.
Anaphylaxis to Patent Blue V. II. A unique IgE‐mediated reaction
Authors: Johansson S. G. O. , Nopp A. , Öman H. , Stahl‐Skov P. , Hunting A. S. , Guttormsen A. B.
Published: October 2009

Background:  Patent Blue V (PBV) is injected in order to map sentinel nodes during cancer staging procedures. Anaphylactic reactions, allegedly IgE antibody mediated, have been reported. The aim of the study was to explore the immunological mechanism of anaphylaxis to PBV.

Methods:  PBV allergen threshold basophil sensitivity, CD‐sens, was performed on cells from nine patients diagnosed as having had adverse reactions to PBV. The mechanisms of the CD‐sens were studied by immunological and immuno‐chemical methods.

Results:  Five of the nine patients had a positive CD‐sens to PBV which was completely eliminated by washing the cells in phosphate buffered saline before allergen challenge. However, the positive CD‐sens was completely reconstituted by incubating the cells in plasma or serum of that patient or the other PBV‐anaphylactic patients for 15 min at room temperature. In some patients the factor mediating CD‐sens was completely or partially destroyed by heating at +56°C for 30 min or being exposed to the low pH used for elution from anti‐Ig columns. A 1000‐fold excess of monoclonal IgE blocked the reconstitution by approximately 50%.

Conclusion:  Anaphylactic reactions to PBV are mediated by IgE antibodies giving a classical CD‐sens reaction. However, the allergenic configuration seems to constitute a structure completely dependent on PBV, as a hapten, linked to a, so far, unknown carrier that seems to be unique for patients having experienced a PBV‐induced reaction. Further studies are needed to characterize the postulated carrier.