bluuebunny/crossref_metadata_2025 huggingface dataset:
HAWAIIAN SKIRT: An F-Box Gene That Regulates Organ Fusion and Growth in Arabidopsis
Authors: González-Carranza Zinnia H. , Rompa Unchalee , Peters Janny L. , Bhatt Anuj M. , Wagstaff Carol , Stead Anthony D. , Roberts Jeremy A.
Published: May 2007

A fast neutron-mutagenized population of Arabidopsis (Arabidopsis thaliana) Columbia-0 wild-type plants was screened for floral phenotypes and a novel mutant, termed hawaiian skirt (hws), was identified that failed to shed its reproductive organs. The mutation is the consequence of a 28 bp deletion that introduces a premature amber termination codon into the open reading frame of a putative F-box protein (At3g61590). The most striking anatomical characteristic of hws plants is seen in flowers where individual sepals are fused along the lower part of their margins. Crossing of the abscission marker, ProPGAZAT:β-glucuronidase, into the mutant reveals that while floral organs are retained it is not the consequence of a failure of abscission zone cells to differentiate. Anatomical analysis indicates that the fusion of sepal margins precludes shedding even though abscission, albeit delayed, does occur. Spatial and temporal characterization, using ProHWS:β-glucuronidase or ProHWS:green fluorescent protein fusions, has identified HWS expression to be restricted to the stele and lateral root cap, cotyledonary margins, tip of the stigma, pollen, abscission zones, and developing seeds. Comparative phenotypic analyses performed on the hws mutant, Columbia-0 wild type, and Pro35S:HWS ectopically expressing lines has revealed that loss of HWS results in greater growth of both aerial and below-ground organs while overexpressing the gene brings about a converse effect. These observations are consistent with HWS playing an important role in regulating plant growth and development.
Identification of a Bipartite Jasmonate-Responsive Promoter Element in the Catharanthus roseus ORCA3 Transcription Factor Gene That Interacts Specifically with AT-Hook DNA-Binding Proteins
Authors: Vom Endt Débora , Soares e Silva Marina , Kijne Jan W. , Pasquali Giancarlo , Memelink Johan
Published: May 2007

Jasmonates are plant signaling molecules that play key roles in defense against certain pathogens and insects, among others, by controlling the biosynthesis of protective secondary metabolites. In Catharanthus roseus, the APETALA2-domain transcription factor ORCA3 is involved in the jasmonate-responsive activation of terpenoid indole alkaloid biosynthetic genes. ORCA3 gene expression is itself induced by jasmonate. By loss- and gain-of-function experiments, we located a 74-bp region within the ORCA3 promoter, which contains an autonomous jasmonate-responsive element (JRE). The ORCA3 JRE is composed of two important sequences: a quantitative sequence responsible for a high level of expression and a qualitative sequence that appears to act as an on/off switch in response to methyl jasmonate. We isolated 12 different DNA-binding proteins having one of four different types of DNA-binding domains, using the ORCA3 JRE as bait in a yeast (Saccharomyces cerevisiae) one-hybrid transcription factor screening. The binding of one class of proteins bearing a single AT-hook DNA-binding motif was affected by mutations in the quantitative sequence within the JRE. Two of the AT-hook proteins tested had a weak activating effect on JRE-mediated reporter gene expression, suggesting that AT-hook family members may be involved in determining the level of expression of ORCA3 in response to jasmonate.
Comparative Cross-Species Alternative Splicing in Plants
Authors: Ner-Gaon Hadas , Leviatan Noam , Rubin Eitan , Fluhr Robert
Published: May 2007

Alternative splicing (AS) can add significantly to genome complexity. Plants are thought to exhibit less AS than animals. An algorithm, based on expressed sequence tag (EST) pairs gapped alignment, was developed that takes advantage of the relatively small intron and exon size in plants and directly compares pairs of ESTs to search for AS. EST pairs gapped alignment was first evaluated in Arabidopsis (Arabidopsis thaliana), rice (Oryza sativa), and tomato (Solanum lycopersicum) for which annotated genome sequence is available and was shown to accurately predict splicing events. The method was then applied to 11 plant species that include 17 cultivars for which enough ESTs are available. The results show a large, 3.7-fold difference in AS rates between plant species with Arabidopsis and rice in the lower range and lettuce (Lactuca sativa) and sorghum (Sorghum bicolor) in the upper range. Hence, compared to higher animals, plants show a much greater degree of variety in their AS rates and in some plant species the rates of animal and plant AS are comparable although the distribution of AS types may differ. In eudicots but not monocots, a correlation between genome size and AS rates was detected, implying that in eudicots the mechanisms that lead to larger genomes are a driving force for the evolution of AS.
Patterns of Selection and Tissue-Specific Expression among Maize Domestication and Crop Improvement Loci
Authors: Hufford Kristina M. , Canaran Payan , Ware Doreen H. , McMullen Michael D. , Gaut Brandon S.
Published: May 2007

The domestication of maize (Zea mays sp. mays) from its wild progenitors represents an opportunity to investigate the timing and genetic basis of morphological divergence resulting from artificial selection on target genes. We compared sequence diversity of 30 candidate selected and 15 reference loci between the three populations of wild teosintes, maize landraces, and maize inbred lines. We inferred an approximately equal ratio of genes selected during early domestication and genes selected during modern crop breeding. Using an expanded dataset of 48 candidate selected and 658 neutral reference loci, we tested the hypothesis that candidate selected genes in maize are more likely to have transcriptional functions than neutral reference genes, but there was no overrepresentation of regulatory genes in the selected gene dataset. Electronic northern analysis revealed that candidate genes are significantly overexpressed in the maize ear relative to vegetative tissues such as maize shoot, leaf, and root tissue. The maize ear underwent dramatic morphological alteration upon domestication and has been a continuing target of selection for maize yield. Therefore, we hypothesize that genes targeted by selection are more likely to be expressed in tissues that experienced high levels of morphological divergence during domestication and crop improvement.
Deletion of Glycine Decarboxylase in Arabidopsis Is Lethal under Nonphotorespiratory Conditions
Authors: Engel Nadja , van den Daele Kirsten , Kolukisaoglu Üner , Morgenthal Katja , Weckwerth Wolfram , Pärnik Tiit , Keerberg Olav , Bauwe Hermann
Published: May 2007

The mitochondrial multienzyme glycine decarboxylase (GDC) catalyzes the tetrahydrofolate-dependent catabolism of glycine to 5,10-methylene-tetrahydrofolate and the side products NADH, CO2, and NH3. This reaction forms part of the photorespiratory cycle and contributes to one-carbon metabolism. While the important role of GDC for these two metabolic pathways is well established, the existence of bypassing reactions has also been suggested. Therefore, it is not clear to what extent GDC is obligatory for these processes. Here, we report on features of individual and combined T-DNA insertion mutants for one of the GDC subunits, P protein, which is encoded by two genes in Arabidopsis (Arabidopsis thaliana). The individual knockout of either of these two genes does not significantly alter metabolism and photosynthetic performance indicating functional redundancy. In contrast, the double mutant does not develop beyond the cotyledon stage in air enriched with 0.9% CO2. Rosette leaves do not appear and the seedlings do not survive for longer than about 3 to 4 weeks under these nonphotorespiratory conditions. This feature distinguishes the GDC-lacking double mutant from all other known photorespiratory mutants and provides evidence for the nonreplaceable function of GDC in vital metabolic processes other than photorespiration.
Monoacylglycerols Are Components of Root Waxes and Can Be Produced in the Aerial Cuticle by Ectopic Expression of a Suberin-Associated Acyltransferase
Authors: Li Yonghua , Beisson Fred , Ohlrogge John , Pollard Mike
Published: May 2007

The interface between plants and the environment is provided for aerial organs by epicuticular waxes that have been extensively studied. By contrast, little is known about the nature, biosynthesis, and role of waxes at the root-rhizosphere interface. Waxes isolated by rapid immersion of Arabidopsis (Arabidopsis thaliana) roots in organic solvents were rich in saturated C18-C22 alkyl esters of p-hydroxycinnamic acids, but also contained significant amounts of both α- and β-isomers of monoacylglycerols with C22 and C24 saturated acyl groups and the corresponding free fatty acids. Production of these compounds in root waxes was positively correlated to the expression of sn-glycerol-3-P acyltransferase5 (GPAT5), a gene encoding an acyltransferase previously shown to be involved in aliphatic suberin synthesis. This suggests a direct metabolic relationship between suberin and some root waxes. Furthermore, when ectopically expressed in Arabidopsis, GPAT5 produced very-long-chain saturated monoacylglycerols and free fatty acids as novel components of cuticular waxes. The crystal morphology of stem waxes was altered and the load of total stem wax compounds was doubled, although the major components typical of the waxes found on wild-type plants decreased. These results strongly suggest that GPAT5 functions in vivo as an acyltransferase to a glycerol-containing acceptor and has access to the same pool of acyl intermediates and/or may be targeted to the same membrane domain as that of wax synthesis in aerial organs.
Phosphatidylinositol 4-Kinase Activation Is an Early Response to Salicylic Acid in Arabidopsis Suspension Cells
Authors: Krinke Ondřej , Ruelland Eric , Valentová Olga , Vergnolle Chantal , Renou Jean-Pierre , Taconnat Ludivine , Flemr Matyáš , Burketová Lenka , Zachowski Alain
Published: May 2007

Salicylic acid (SA) has a central role in defense against pathogen attack. In addition, its role in such diverse processes as germination, flowering, senescence, and thermotolerance acquisition has been documented. However, little is known about the early signaling events triggered by SA. Using Arabidopsis (Arabidopsis thaliana) suspension cells as a model, it was possible to show by in vivo metabolic phospholipid labeling with 33Pi that SA addition induced a rapid and early (in few minutes) decrease in a pool of phosphatidylinositol (PI). This decrease paralleled an increase in PI 4-phosphate and PI 4,5-bisphosphate. These changes could be inhibited by two different inhibitors of type III PI 4-kinases, phenylarsine oxide and 30 μ  m wortmannin; no inhibitory effect was seen with 1 μ  m wortmannin, a concentration inhibiting PI 3-kinases but not PI 4-kinases. We therefore undertook a study of the effects of wortmannin on SA-responsive transcriptomes. Using the Complete Arabidopsis Transcriptome MicroArray chip, we could identify 774 genes differentially expressed upon SA treatment. Strikingly, among these genes, the response to SA of 112 of them was inhibited by 30 μ  m wortmannin, but not by 1 μ  m wortmannin.
Arabidopsis VIRE2 INTERACTING PROTEIN2 Is Required for Agrobacterium T-DNA Integration in Plants
Authors: Anand Ajith , Krichevsky Alexander , Schornack Sebastian , Lahaye Thomas , Tzfira Tzvi , Tang Yuhong , Citovsky Vitaly , Mysore Kirankumar S.
Published: May 2007

Agrobacterium tumefaciens–mediated genetic transformation is an efficient tool for genetic engineering of plants. VirE2 is a single-stranded DNA binding Agrobacterium protein that is transported into the plant cell and presumably protects the T-DNA from degradation. Using a yeast two-hybrid system, we identified Arabidopsis thaliana VIRE2-INTERACTING PROTEIN2 (VIP2) with a NOT domain that is conserved in both plants and animals. Furthermore, we provide evidence supporting VIP2 interaction with VIP1, a basic domain/leucine zipper motif–containing protein required for nuclear import and integration of T-DNA. Virus-induced gene silencing of VIP2 in Nicotiana benthamiana and characterization of the Arabidopsis vip2 mutant (At vip2) demonstrate that VIP2 is required for Agrobacterium-mediated stable transformation but not for transient transformation. Assays based upon a promoter-trap vector and quantification of T-DNA integration further confirmed VIP2 involvement in T-DNA integration. Interestingly, VIP2 transcripts were induced to a greater extent over prolonged periods after infection with a T-DNA transfer-competent Agrobacterium strain compared with the transfer-deficient Agrobacterium strain. Transcriptome analyses of At vip2 suggest that VIP2 is likely a transcriptional regulator, and the recalcitrancy to transformation in At vip2 is probably due to the combination of muted gene expression response upon Agrobacterium infection and repression of histone genes resulting in decreased T-DNA integration events.
CYP703 Is an Ancient Cytochrome P450 in Land Plants Catalyzing in-Chain Hydroxylation of Lauric Acid to Provide Building Blocks for Sporopollenin Synthesis in Pollen
Authors: Morant Marc , Jørgensen Kirsten , Schaller Hubert , Pinot Franck , Møller Birger Lindberg , Werck-Reichhart Danièle , Bak Søren
Published: May 2007

CYP703 is a cytochrome P450 family specific to land plants. Typically, each plant species contains a single CYP703. Arabidopsis thaliana CYP703A2 is expressed in the anthers of developing flowers. Expression is initiated at the tetrad stage and restricted to microspores and to the tapetum cell layer. Arabidopsis CYP703A2 knockout lines showed impaired pollen development and a partial male-sterile phenotype. Scanning electron and transmission electron microscopy of pollen from the knockout plants showed impaired pollen wall development with absence of exine. The fluorescent layer around the pollen grains ascribed to the presence of phenylpropanoid units in sporopollenin was absent in the CYP703A2 knockout lines. Heterologous expression of CYP703A2 in yeast cells demonstrated that CYP703 catalyzes the conversion of medium-chain saturated fatty acids to the corresponding monohydroxylated fatty acids, with a preferential hydroxylation of lauric acid at the C-7 position. Incubation of recombinant CYP703 with methanol extracts from developing flowers confirmed that lauric acid and in-chain hydroxy lauric acids are the in planta substrate and product, respectively. These data demonstrate that in-chain hydroxy lauric acids are essential building blocks in sporopollenin synthesis and enable the formation of ester and ether linkages with phenylpropanoid units. This study identifies CYP703 as a P450 family specifically involved in pollen development.
TIME FOR COFFEEEncodes a Nuclear Regulator in theArabidopsis thalianaCircadian Clock
Authors: Ding Zhaojun , Millar Andrew J. , Davis Amanda M. , Davis Seth J.
Published: May 2007

The plant circadian clock is required for daily anticipation of the diurnal environment. Mutation in Arabidopsis thaliana TIME FOR COFFEE (TIC) affects free-running circadian rhythms. To investigate how TIC functions within the circadian system, we introduced markers for the evening and morning phases of the clock into tic and measured evident rhythms. The phases of evening clock genes in tic were all advanced under light/dark cycles without major expression level defects. With regard to morning-acting genes, we unexpectedly found that TIC has a closer relationship with LATE ELONGATED HYPOCOTYL (LHY) than with CIRCADIAN CLOCK ASSOCIATED1, as tic has a specific LHY expression level defect. Epistasis analysis demonstrated that there were no clear rhythms in double mutants of tic and evening-acting clock genes, although double mutants of tic and morning-acting genes exhibited a similar free-running period as tic. We isolated TIC and found that its mRNA expression is continuously present over the diurnal cycle, and the encoded protein appears to be strictly localized to the nucleus. Neither its abundance nor its cellular distribution was found to be clock regulated. We suggest that TIC encodes a nucleus-acting clock regulator working close to the central oscillator.
The Two AGPase Subunits Evolve at Different Rates in Angiosperms, yet They Are Equally Sensitive to Activity-Altering Amino Acid Changes When Expressed in Bacteria
Authors: Georgelis Nikolaos , Braun Edward L. , Shaw Janine R. , Hannah L. Curtis
Published: May 2007

The rate of protein evolution is generally thought to reflect, at least in part, the proportion of amino acids within the protein that are needed for proper function. In the case of ADP-glucose pyrophosphorylase (AGPase), this premise led to the hypothesis that, because the AGPase small subunit is more conserved compared with the large subunit, a higher proportion of the amino acids of the small subunit are required for enzyme activity compared with the large subunit. Evolutionary analysis indicates that the AGPase small subunit has been subject to more intense purifying selection than the large subunit in the angiosperms. However, random mutagenesis and expression of the maize (Zea mays) endosperm AGPase in bacteria show that the two AGPase subunits are equally predisposed to enzyme activity-altering amino acid changes when expressed in one environment with a single complementary subunit. As an alternative hypothesis, we suggest that the small subunit exhibits more evolutionary constraints in planta than does the large subunit because it is less tissue specific and thus must form functional enzyme complexes with different large subunits. Independent approaches provide data consistent with this alternative hypothesis.
Increasing Plasma Membrane Phosphatidylinositol(4,5)Bisphosphate Biosynthesis Increases Phosphoinositide Metabolism inNicotiana tabacum
Authors: Im Yang Ju , Perera Imara Y. , Brglez Irena , Davis Amanda J. , Stevenson-Paulik Jill , Phillippy Brian Q. , Johannes Eva , Allen Nina S. , Boss Wendy F.
Published: May 2007

A genetic approach was used to increase phosphatidylinositol(4,5)bisphosphate [PtdIns(4,5)P2] biosynthesis and test the hypothesis that PtdInsP kinase (PIPK) is flux limiting in the plant phosphoinositide (PI) pathway. Expressing human PIPKIα in tobacco (Nicotiana tabacum) cells increased plasma membrane PtdIns(4,5)P2 100-fold. In vivo studies revealed that the rate of 32Pi incorporation into whole-cell PtdIns(4,5)P2 increased >12-fold, and the ratio of [3H]PtdInsP2 to [3H]PtdInsP increased 6-fold, but PtdInsP levels did not decrease, indicating that PtdInsP biosynthesis was not limiting. Both [3H]inositol trisphosphate and [3H]inositol hexakisphosphate increased 3-and 1.5-fold, respectively, in the transgenic lines after 18 h of labeling. The inositol(1,4,5)trisphosphate [Ins(1,4,5)P3] binding assay showed that total cellular Ins(1,4,5)P3/g fresh weight was >40-fold higher in transgenic tobacco lines; however, even with this high steady state level of Ins(1,4,5)P3, the pathway was not saturated. Stimulating transgenic cells with hyperosmotic stress led to another 2-fold increase, suggesting that the transgenic cells were in a constant state of PI stimulation. Furthermore, expressing Hs PIPKIα increased sugar use and oxygen uptake. Our results demonstrate that PIPK is flux limiting and that this high rate of PI metabolism increased the energy demands in these cells.
The use of single‐use devices in anaesthesia: balancing the risks to patient safety*
Authors: Rowley E. , Dingwall R.
Published: May 2007

Single‐use devices are designed, manufactured and sold to be used once and then discarded. This paper addresses growing concerns about the quality of some devices. Single‐use devices, manufactured at a lower cost to justify their disposal, are perceived to have a lesser efficacy, which may threaten patient safety through iatrogenic harm. There is, in addition, growing scepticism about the actual risk of contracting variant Creutzfeldt‐Jakob disease and other blood‐borne diseases from reused surgical instruments. Interview data suggests that when choosing to use a single‐use device, clinicians balance concerns about the risk of infection against those about the risk of injury. However, despite reservations about induced harm and the unknown risk of an iatrogenic disease, most clinicians would want single‐use devices used on themselves and their family if they were patients.
Double trouble for grasshopper molecular systematics: intra‐individual heterogeneity of both mitochondrial 12S‐valine‐16S and nuclear internal transcribed spacer ribosomal DNA sequences in Hesperotettix viridis (Orthoptera: Acrididae)
Authors: SWORD GREGORY A. , SENIOR LAURA B. , GASKIN JOHN F. , JOERN ANTHONY
Published: May 2007

Abstract Hesperotettix viridis grasshoppers (Orthoptera: Acrididae: Melanoplinae) exhibit intra‐individual variation in both mitochondrial 12S‐valine‐16S and nuclear internal transcribed spacer (ITS) ribosomal DNA sequences. These findings violate core assumptions underlying DNA sequence data obtained via polymerase chain reaction (PCR) amplification for use in molecular systematics investigations. Undetected intra‐individual variation of this sort can confound phylogenetic analyses at a range of taxonomic levels. The use of a DNA extraction protocol designed to enrich mitochondrial DNA as well as an initial long PCR of approximately 40% of the grasshopper mitochondrial genome failed to control for the presence of paralogous mitochondrial DNA‐like sequences within individuals. These findings constitute the first demonstration of intra‐individual heterogeneity in mitochondrial DNA‐like sequences in the grasshopper subfamily, Melanoplinae, and only the second report of intra‐individual variation in nuclear ITS ribosomal DNA sequences in grasshoppers. The fact that intra‐individual variation was detected in two independent DNA marker sets in the same organism strengthens the notion that the orthology of PCR‐derived DNA sequences should be examined thoroughly prior to their use in molecular phylogenetic analyses or as DNA barcodes.
PRECLINICAL STUDY: Perinatal alcohol exposure enhances nocistatin levels in adulthood
Authors: Tekes Kornélia , Hantos Mónika , Gyenge Melinda , Csaba Gyorgy
Published: May 2007

In earlier experiments perinatal hormonal imprinting by alcohol decreased the hormone content of immune cells for life. In the present study, both a single day (15% on the third postnatal day) and a long‐term treatment schedule of alcohol exposure (3% for 21 days) of dams during lactation significantly (P < 0.01) enhanced endogenous levels of nocistatin in the blood plasma as well as in the cerebrospinal fluid of the offspring, measured in 3‐month‐old rats. Our data suggest that alcohol consumption during lactation can cause a life‐long influence on nocistatin levels in the offspring and most likely modify nocistatin‐related functions such as pain tolerance.
PRECLINICAL STUDY: Effect of cannabinoids on platelet serotonin uptake
Authors: Velenovská Marie , Fišar Zdeněk
Published: May 2007

Serotonin is involved in many of the same processes affected by cannabinoids; therefore, we investigated in vitro and in vivo effects of these drugs on the function of serotonin transporter. The effect of Δ9‐tetrahydrocannabinol (Δ9‐THC), endocannabinoid anandamide and synthetic cannabinoid receptor agonist WIN 55,212‐2 on platelet serotonin uptake and membrane microviscosity was examined in 19 marijuana smokers and 20 controls. (1) Serotonin uptake was inhibited at higher doses of Δ9‐THC (IC50 = 139 µmol/l), anandamide (IC50 = 201 µmol/l) or WIN 55,212‐2 (IC50 = 17.4 µmol/l); the inhibition was found non‐competitive. Δ9‐THC, anandamide and WIN 55,212‐2 produced different effects on the membrane microviscosity. (2) Maximal velocity of platelet serotonin uptake was significantly increased in a group of chronic marijuana smokers suffering impairment of cognitive functions when compared with controls. Opposite effect of marijuana smoking on the serotonin uptake efficiency was observed in males beside females. In summary, this study provides evidence that (1) Activity of serotonin transporter is acutely affected by cannabinoids at relatively high drug concentrations; this effect is indirect and can be partially accounted for the changes in the membrane microviscosity. (2) Increase of maximal velocity of the serotonin uptake could be understood as adaptation change in the serotonergic system induced by chronic cannabis use. A hypothesis was supported that lowered serotonin uptake may reflect a gender‐related differences in effects of psychoactive cannabinoids.
CLINICAL STUDY: Physical health problems among patients seeking treatment for alcohol use disorders: a study in six European cities
Authors: Gossop Michael , Neto Domingos , Radovanovic Mirjana , Batra Anil , Toteva Sonya , Musalek Michael , Skutle Arvid , Goos Cees
Published: May 2007

The present study investigates physical health problems among patients with alcohol use disorders at alcohol treatment agencies in six European cities. The sample comprised 315 patients with a primary alcohol use disorder. Data were collected at admission to treatment using a structured research protocol, and ratings were made by a medically qualified physician subsequent to a physical examination of the patient. Physical health problems were extremely common: 79% of the sample had at least one problem, and 59% had two or more problems. Health problems were often serious, and 60% had at least one health problem that required treatment. The most common problems were gastrointestinal and liver disorders, but about a quarter of the sample had cardiovascular or neurological problems. Frequency of drinking, duration of alcohol use disorder, and severity of alcohol dependence were associated with increased physical morbidity. Current smoking status and age were also associated with poorer physical health. Older drinkers had more physical health problems although they were less severely alcohol dependent than their younger counterparts. The high prevalence of physical health problems among problem drinkers provides opportunities of screening for alcohol use disorders not only in specialist alcohol treatment services but also in other health‐care settings. It is recommended that alcohol treatment agencies should provide a full routine health screen of patients at admission to treatment with provision or referral to appropriate treatment.
PRECLINICAL STUDY: Morphine withdrawal decreases responding reinforced by sucrose self‐administration in progressive ratio
Authors: Zhang Dengke , Zhou Xuhui , Wang Xuyi , Xiang Xiaojun , Chen Hongxian , Hao Wei
Published: May 2007

Previous studies have shown that withdrawal from psychostimulant drugs such as d‐amphetamine or methamphetamine decreases motivation to work for a natural reinforcement, which is thought to be associated with the withdrawal‐induced depressive state and hypofunction of the mesolimbic dopamine system. However, to our knowledge, studies exploring the effect of morphine withdrawal on motivation for a natural reinforcement are lacking. The purpose of the present study was to examine whether motivation to work for a natural reinforcement changes during morphine withdrawal. Three groups of male Sprague–Dawley rats were trained to respond on a nose poke for a 4% sucrose solution under a progressive ratio schedule and were subsequently administered a 10‐day regimen of injection of high or low dose of morphine or saline. Their duration of break point and withdrawal symptoms were assessed. The finding showed that break points were significantly reduced on day 1 and persisted to at least day 10 of withdrawal without change in locomotor activity. There were hardly any differences bear mentioning when comparing the magnitude of the decrease between the high‐ and the low‐dose group, whereas the withdrawal scales were significant greater in the high‐dose group than in the low‐dose group. The results suggest that the morphine withdrawal resulted in decreased motivation to obtain the natural reinforcement. The progressive ratio procedure may be a useful technique for evaluation of changes in motivation for natural reinforcing stimuli following withdrawal from opiates.
Pulmonary tuberculosis in patients treated with inhaled beclomethasone
Authors: Shaikh W. A.
Published: May 2007

Inhaled beclomethasone dipropionate (BDP) has been used with few side effects in the treatment of bronchial asthma for 2 decades. Until now the manifestation of tuberculosis (TB) in patients on inhaled BDP has not been reported. Eight patients with allergic asthma, of a total of 548 asthmatics (1.46%) seen over a 2‐year period, developed active TB following the use of inhaled BDP. All were sputum‐positive for acid‐fast bacilli (AFB) on smear and/or culture, all responded well to a combination of anti‐TB drugs, and none showed evidence of immunological or pituitary‐adrenal suppression. Two patients agreed to a repeat administration of BDP; both developed TB again within 2 weeks and are again on anti‐TB treatment.
Modulation of eicosanoid and histamine release from human dispersed lung cells by terfenadine
Authors: Campbell A. M. , Chanez P. , Marty‐Ané C. , Albat B. , Bloom M. , Michel F. B. , Godard P. , Bousquet J.
Published: May 2007

Terfenadine is an H1‐blocker that may have antiallergic properties. A study was carried out to examine the ability of terfenadine to inhibit the release of histamine and arachidonic‐acid‐derived mediators from human lung cells. Cells were dispersed from fresh human lung tissue obtained from tour accident victims whose hearts were donated for transplantation and four lung cancer resections. Cells were dispersed by enzymatic digestion with type XIV protease and chymopapain, and this resulted in a cell population containing approximately 5% mast cells. The remaining cells were mainly macrophages. The cells were challenged with anti‐IgE at a 1/1000 dilution. Cells were challenged without terfenadine and after a preincubation of 0.1, 1, and 10 umol terfenadine. The release of PGD2 and LTC4/D4 was assessed with an EIA. Histamine was assayed by RIA with a monoclonal antibody against acylated histamine.

A release of both eicosanoids and histamine was observed m all experiments. An inhibition of eicosanoids was observed at both 1 and 10 μmol terfenadine (median percentage of inhibition of PGD2: 38.00 ± 15.65 and 56,00 ± 13,12; median percentage of inhibition of LTC4/D4: 37.5 ± 19,80 and 52.5 ± 26.8). On the other hand, histamine release was not blocked by terfenadine.

Terfenadine inhibits, in a dose‐dependent manner, the release of eicosanoids after challenge of dispersed lung cells by anti‐IgE, and this effect may have some clinical relevance.
Investigation of basophil cell proteinase release in progressive systemic sclerosis
Authors: Seres T. , Nagy Z. , Czirják L.
Published: May 2007

The aim of this study was to evaluate the function of basophil granulocytes in 37 patients with progressive systemic sclerosis (PSS). The in vitro proteinase release test using Chromozyme TH as a chromogenic substrate was performed to measure the basophil cell releasability. Anti‐IgE, calcium‐ionophore (A 23187) and N‐formyl‐methyonil‐leueyl‐phenylalanine were used as activatory agents. Our results showed that the reactivity pattern to those agents did not differ in PSS compared to controls. Patients whose basophil cells release proteinases to all of the 3 activators tended to have more severe organ symptoms compared to the rest of the patients, while teleangiectasia was significantly less frequently (p<0.02) found in cases with proteinase release. These findings suggest that there may be a certain relationship between basophil function and severity of the PSS.
Effect of dry cleaning on mite allergen levels in blankets
Authors: Vandenhove T. , Soler M. , Birnbaum J. , Charpin D. , Vervloet D.
Published: May 2007

Since one of the greatest reservoirs of allergens is the blanket, we assessed mite allergen levels in dust collected from five blankets by vacuuming before and after dry cleaning with perchlorethylene and compared the results with five control blankets. Assays with monoclonal antibodies showed that group I (Der p I and Der f I) mite allergen levels per g dust were 78%, lower after dry cleaning. Group 1 allergen levels per m2 of dry‐cleaned blankets were 98% lower. RAST inhibition showed that total allergen levels decreased 70% after dry cleaning. Mite allergens were not denatured by perchlorethylene. The effect of dry cleaning resulted from physical washing out of dust and allergens.
Pulmonary function in adolescents with childhood asthma
Authors: Laursen E. Mosfeldt , Hansen K. Kaae , Backer V. , Bach‐Mortensen N. , Prahl P. , Koch C.
Published: May 2007

The aim of this study was to determine the pulmonary function in former and present asthmatics. We examined 77 persons aged 12–24 years, classified into four groups: 1) healthy subjects (controls) (n= 19), 2) former asthmatics (n= 19), 3) present mild asthmatics (n= 20), and 4) present severe asthmatics (n= 19). Although exhibiting no respiratory symptoms, former asthmatics had reduced airflow values measured by FEV, (median (range) 89.7 (83–99) vs 101.4 (91–110)) and MEF25 (76.5 (68–94) vs 103.0 (97–124)), as compared with controls. Furthermore, former asthmatics had significantly increased PEF variability, as compared with controls, whereas no significant differences were found in static lung parameters, i.e. total lung capacity and residual volume, as compared with controls. In conclusion, former asthmatics, although now exhibiting no respiratory symptoms, were found to have obstructive airflow limitation, increased bronchial responsiveness, and normal lung volumes.
Deflazacort protects against late‐phase but not early‐phase reactions induced by the allergen‐specific conjunctival provocation test
Authors: Ciprandi G. , Buscaglia S. , Pesce G. P. , Iudice A. , Bagnasco M. , Canonica G. W.
Published: May 2007

The protective effects of deflazacort against the inflammation that follows the conjunctival provocation test (CPT) by specific allergen were assessed in 24 patients with rhinoconjunctivitis caused by Parietaria judaica in a double‐blind study. After a screening CPT, patients were randomized into four treatment groups, each being given deflazacort (oral tablets) at 6, 30, and 60 mg once daily, or matching placebo, for 3 d, outside the pollen season. Clinical evaluation (itching, hyperemia, lacrimation, and swelling of eyelids) and cytologic assessment (number of inflammatory cells in conjunctival scraping and evaluation of ICAM (intercellular adhesion molecule)‐1/CD54 expression on epithelial cells) were performed at base line, 30 min (early‐phase reaction (EPR), 6 h and 24 h (late‐phase reaction (LPR)) after specific CPT, and before and after treatment. Neither the EPR clinical reactions nor the EPR total number of inflammatory cells was modified by deflazacort. However, the LPR clinical effects were significantly reduced by deflazacort at 30 or 60mg/d (P<0.01), as compared with placebo. The total number of inflammatory cells during LPR was significantly reduced by deflazacort at 30 or 60 nig/d ((P<0.01), as compared with placebo. Furthermore, CD54 expression was significantly reduced by deflazacort at 30 or 60 mg/d both in the EPR ((P<0.01) and LPR ((P<0.01), as compared with placebo. None of the studied indicators were modified at the 6 mg/d dose. This study shows that deflazacort has a highly protective action against clinical and cellular LPR effects induced by the specific CPT, In addition, deflazacort markedly reduces CD54 expression on the conjunctival epithelium during both EPR and LPR.
Characterization and partial purification of the Croatian national standard Dermatophagoides pteronyssinus allergen extract
Authors: Tresščcec A. , Kolevska T. , Cvorišsšcec B. , Krnić B. , Stipić‐Marković A. , Tudman Z. , Dekaris D.
Published: May 2007

Lyophilized Dermatophagoides pteronyssinus (Der p) allergen extract (AE) and partially purified Der p extract (PAE) Were Prepared and characterized. Partial purification of AE was performed by gel filtration on Sephadex G‐1OO and Sephacryl S‐300. Crossed immunoelectrophoresis (CIE) disclosed the same precipitating lines in AE and PAE preparations. The relative potencies of AE and PAE were determined and compared with the WHO International Standard for Der p by the RAST inhibition method. The potencies were 6.5 × 105 IU and 1.5 × 106 IU, respectively. Biologic standardization by quantitative skin testing was performed with AE (20 selected patients) and PAE (12 patients). Median Ch was calculated by linear regression analysis (log‐log model). One ampoule of AE contained 65 300 BU and 1 ml (vial) of PAE contained 166000 BU. Der p AE could serve as a Croatian national standard for further production of Der p allergenic extracts.
Plasma histamine and bronchial reactivity in allergic asthma
Authors: Corhay J. L. , Bury Th. , Louis R. , Radermecker M. F.
Published: May 2007

Histamine is an important mediator of allergic inflammation and bronchial hyperresponsiveness (BHR), a hallmark of asthma. Studies on the relationship between plasma histamine and BHR in allergic asthmatic patients have yielded controversial results. We therefore measured plasma histamine and bronchial reactivity in 30 nonsmoker volunteers taking no medication. Eleven were normal subjects; 19 were stable, mildly allergic asthmatic patients. Venous blood was taken to measure blood cells and basal plasma histamine by radioimmunoassay. After blood sampling, all subjects underwent a measurement of PC20M (concentration of methacholine causing a 20% fall in FEV1). Mean plasma histamine levels were 0.21 ± 0.1 ng/ml and 0.44 ± 0.3 ng/ml in normal and asthmatic subjects, respectively (P<0.05). We found a significant increase of blood eosinophils and basophils in asthmatic patients, and a positive correlation between plasma histamine and circulating basophils. PC20M was greater than 16 mg in normal volunteers, and mean PC20M was 2.1 ± 2 mg/ml in asthmatic patients. PC20M did not correlate with plasma histamine levels, but it did so negatively with blood eosinophils. The increased plasma histamine concentration in mildly atopic asthmatic patients might be a consequence of the high basophil releasability of atopies and the higher basophil counts in allergic asthma. Plasma histamine is thus unlikely to be a determinant of BHR in asthma.
Mite‐allergen exposure and spontaneous histamine release in monosensitive and polysensitive mite‐allergic patients
Authors: Weyer A. , de Blay F. , Hirth C. , Marchand F. , Tekaia F. , Pauli G. , David B.
Published: May 2007

Forty‐three patients allergic to mites and suffering mainly from asthma were recruited: 25 were mite‐monosensitive and 18 were polysensitive, as determined by skin tests and specific serum IgE determinations with various allergens. In vitro spontaneous histamine release (SHR) by washed blood basophils was measured once or several times for each patient. Throughout this study, the mean periods of high and low mite‐allergen exposure were defined on the basis of relative indoor humidity and temperature data. For the mite‐monosensitive patients, there was a significant increase in mean SHR during the season of high mite‐allergen exposure as compared to the months of lower mite‐allergen presence (P < 0.002). No significant difference between mean SHR values was observed when comparing the monosensitive group during the season of high mite‐allergen exposure with polysensitive patients (allergic to mite and pollen) during the period of exposure to both allergens. Differences in mean SHR reported here emphasize the positive relationship between intense allergen exposure and the in vitro SHR increase in blood basophils of mite‐allergic patients.
Does guanine concentration in house‐dust samples reflect house‐dust mite exposure?
Authors: Hallas T.E. , Yi Xue , Schou C.
Published: May 2007

Using specific ELISA extracts, we analyzed samples of pure mite cultures and house dust for group I allergens from Dermatophagoides pteronyssinus, D. farinae, and D. microceras. Correspondingly, we measured the concentration of guanine and xanthine by rigorous chemical methods. The pure mite cultures were used to correiate allergen levels with guanine levels. The house‐dust samples one‐sidedly contained more guanine than expected. This suggests that considerable amounts of guanine originate from non‐house‐dust mite sources and implies that guanine quantifications are of limited clinical value.
Prediction of mite allergen levels by guanine measurements in house‐dust samples
Authors: Quoix E. , Mao J. Le , Hoyet C. , Pauli G.
Published: May 2007

We studied the sensitivity and specificity of guanine environmental tests in the evaluation of the two mite allergen levels, i.e. 2 üg/g and 10 üg/g of Der p I and Der f I, considered to be risk factors for sensitization or for the development of acute asthma. We examined 239 house‐dust samples for Der p I and Der f I levels (ELISA) and guanine contents (semiquantitative guanine test and quantitative assays). All house‐dust samples with class 2 or 3 guanine tests contained more than 2 üg/g of Der p I and Der f I. The probability that house‐dust samples of class 2 contained more than 10 üg/g of mite allergens was 88%; it was 100% for house‐dust samples of class 3. The probability that a house‐dust sample of class 0 contained less than 2 üg/g of mite allergen was 87%. For each level of mite allergen, a ROC curve was constructed with the true positive rates and the false positive rates calculated by different cutoffs of guanine concentration. The cutoff point which gave the best compromise between sensitivity (76%) and specificity (89%) was 2100 üg/g for the threshold of 2 üg/g of Der p I and Der f I. For detection of a mite allergen > 10 ųg, a guanine content of 3000 üg/g gave the best compromise between sensitivity (86%) and specificity (93%). In conclusion, the guanine test represents a satisfactory environmental test, inexpensive and simple, for predicting mite allergen levels. Semiquantitative and quantitative guanine measurements are more accurate in predicting an exposure level of 10 üg/g than of the level 2 üg/g of Der p I and Der f I.
Localized periorbital edema induced by aspirin
Authors: Katz Y. , Goldberg N. , Kivity S.
Published: May 2007

We documented localized periorbital edema in two patients with aspirin sensitivity without underlying chronic urticaria. The reaction developed 30 min after ingestion of 62.5 and 125 mg of aspirin, respectively. No systemic symptoms were observed. Other NSAIDs did not induce symptoms. These patients were able to tolerate doses of aspirin after pretreatment with terfenadine.

These observations suggest that histamine plays a central role in aspirin‐induced skin reaction. Despite the fact that terfenadine blocks the drug‐induced reaction, this protocol should be used with caution and only where there is no feasible alternative to aspirin.
Assessment of the efficacy and safety of three dose levels of cetirizine given once daily in children with perennial allergic rhinitis
Authors: Jobst S. , van den Wijngaart W. , Schubert A. , van de Venne H.
Published: May 2007

The present study compared the efficacy and safety of three dose levels of cetirizine (2.5. 5, and 10 mg) once a day with placebo over 14 days in 6–12‐year‐old children with perennial allergic rhinitis. The design was a double‐blind, randomized, multicenter, parallel‐group study. Five symptoms (sneezing, nasal discharge, nasal obstruction, nasal pruritus, and ocular pruritus) were rated according to severity by investigators at the visits and daily by patients. Eighty‐three patients were randomized to placebo, 84 to 2.5 mg cetirizine, 85 to 5 mg cetirizine, and 76 to 10 mg cetirizine. Groups were comparable at inclusion. The primary efficacy variable was the percentage of days with no or only mild symptoms: at all doses, cetirizine appeared to be more effective than placebo, but a significant difference was reached only in the 10‐mg group (difference in medians of 22%;P= 0.016). The test of linearity was significant (P= 0.026) for the percentage of asymptomatic days. The investigators’ assessments at each visit scored the symptoms in the placebo group higher, i.e., more severe, than in the active groups, the 10‐mg dose causing the greatest reduction in symptoms. Adverse events were infrequent and generally mild or moderate in severity. It was concluded that cetirizine at a 10‐mg, once daily dose could be used to treat effectively 6–12‐year‐old children with perennial allergic rhinitis.
Topical sodium cromoglycate (Opticrom®) relieves ongoing symptoms of allergic conjunctivitis within 2 minutes
Authors: Montan P. , Zetterström O. , Eliasson E. , Strömquist L.‐H.
Published: May 2007

Montan P, Zetterström O, Eliasson E, Strömquist L‐H. Topical sodium cromoglycate (Opticrom®) relieves ongoing symptoms of allergic conjunctivitis within 2 minutes.

Twelve patients with seasonal allergic conjunctivitis caused by either birch or timothy grass pollen were challenged out of season in both eyes, with the relevant allergen. Itching, redness, swelling, and tearing were assessed on a 0–3 scale. When a positive reaction, i.e., at least 4 points, had occurred in both eyes, one drop of 2% sodium cromoglycate was administered to one eye and one drop of placebo to the other eye in each patient, in a double‐blind, randomized fashion. Symptoms were then assessed after 2, 10, 30, and 60 min. Sodium cromoglycate was statistically better than placebo in reducing the symptoms at 2, 10, and 30 min after the treatment had been administered, showing that topical application of 2% sodium cromoglycate can quickly relieve ongoing symptoms of allergic inflammation in the eve.
Raccoon epithelium – a new allergen source
Authors: Stöger P. , Schmid‐Grendelmeier P. , Johansson S. G. O. , Wüthrich B.
Published: May 2007

We report the case of a 39‐year‐old asthmatic patient who developed acute rhinoconjunctivitis, Quincke's edema, and asthma after repeated exposure to a raccoon (Procyon lotor) which a friend kept at home. Skin prick and scratch tests with native raccoon hair and epithelium, as well as RAST, were clearly positive. Elimination of contact with the raccoon and avoiding the home of his friend led to a cessation of the patient's acute exacerbations.
Adverse reactions to food additives in children with atopic symptoms
Authors: Fuglsang G. , Madsen C. , Halken S. , Jørgensen M. , Østergaard P. A. , Østerballe O.
Published: May 2007

In a multicenter study conducted at four Danish hospital pediatric departments, the parents of 472 consecutive children were informed of this project to determine the incidence of intolerance of food additives among children referred to an allergy clinic with symptoms of asthma, atopic dermatitis, rhinitis, or urticaria. After a 2‐week period on an additive‐free diet, the children were challenged with the eliminated additives. The food additives investigated were coloring agents, preservatives, citric acid, and flavoring agents. Carbonated “lemonade” containing the dissolved additives was used for the open challenge. Two doses were used: a low dose and a 10‐fold higher dose. Gelatin capsules were used for a double‐blind challenge. The children were 4–15 years old, and they were attending an outpatient pediatric clinic for the first time. Of the 379 patients who entered the study, 44 were excluded and 335 were subjected to open challenge. A total of 23 children developed positive reactions after the open challenge. Sixteen of these patients accepted the double‐blind challenge, and six showed a positive reaction to preservatives (atopic dermatitis, asthma, rhinitis), coloring agents (atopic dermatitis, asthma, urticaria, gastrointestinal symptoms), and citric acid (atopic dermatitis, gastrointestinal symptoms). The incidence of intolerance of food additives was 2% (6/335), as based on the double‐blind challenge, and 7% (23/335), as based on the open challenge with lemonade. Children with atopic skin symptoms had a statistically increased risk of a positive reaction. This may have consequences for the future clinical investigation of children with atopic cutaneous symptoms.