bluuebunny/crossref_metadata_2025 huggingface dataset:
GABAB Receptor‐Activation Inhibits GABAergic Synaptic Transmission in Parvocellular Neurones of Rat Hypothalamic Paraventricular Nucleus
Authors: Liu X. , Tribollet E. , Raggenbass M.
Published: December 2005

The paraventricular nucleus of the hypothalamus contains three classes of neurones: (i) magnocellular and (ii) parvocellular neurosecretory neurones and (iii) nonendocrine projection neurones. The present study aimed to determine whether functional GABAB receptors are present on axon terminals that synapse with parvocellular neurosecretory and nonendocrine paraventricular neurones and to determine how activation of GABAB receptors control GABAergic input to these neurones. Whole‐cell recordings were performed in coronal hypothalamic slices of the rat containing the paraventricular nucleus. GABAA receptor–mediated inhibitory postsynaptic currents (i.p.s.c.) were isolated pharmacologically in the presence of antagonists of glutamatergic ionotropic receptors. We found that baclofen, an agonist of GABAB receptors, decreased the frequency of spontaneous and miniature i.p.s.c. It also decreased the amplitude of evoked i.p.s.c. These effects were suppressed by CGP55845A, a competitive antagonist of GABAB receptors. CGP55845A also increased the frequency of miniature i.p.s.c. and the amplitude of evoked i.p.s.c., suggesting that, in physiological conditions, presynaptic GABAB receptors exert a tonic inhibition on GABA release. Baclofen had no effect on GABA‐evoked postsynaptic currents, suggesting that the baclofen‐dependent suppression of GABAergic i.p.s.c. was exclusively due to a presynaptic action of the agonist. Our data indicate that GABAB receptors are present on axon terminals of GABAergic presynaptic neurones contacting parvocellular neurosecretory and nonendocrine paraventricular neurones, and suggest that GABAB receptors exert a tonic inhibition of GABA release from GABAergic terminals. Activation of these receptors causes disinhibition of parvocellular neurosecretory and nonendocrine paraventricular neurones.
Influence of hydrogeographic history and hybridization on the distribution of genetic variation in the pupfishesCyprinodon atrorusandC. bifasciatus
Authors: CARSON EVAN W. , DOWLING THOMAS E.
Published: December 2005

The evolutionary importance of hybridization in animals has been subject of much debate. In this study, we examined the influence of hydrogeographic history and hybridization on the present distribution of nuclear and mitochondrial DNA variation in two pupfish species,Cyprinodon atrorusandCyprinodon bifasciatus. Results presented here indicate that there has been limited introgression of nuclear genes; however, mtDNA introgression has been substantial, with complete replacement of theC.bifasciatusmitochondrial genome by that ofC.atrorus. Subsequent to this replacement, there has been diversification of mitochondrial haplotypes along major geographic regions in the basin. Evidence was also found that mitochondrial replacement follows a predictable, cyclical pattern in this system, with isolation and diversification followed by re‐contact and replacement ofC.bifasciatusmitochondrial haplotypes by those ofC.atrorus. This pattern is best explained by a combination of a numeric bias towardsC.atrorusand mating site selection rather than selection forC.atrorusmitochondrial genome. These results demonstrate the important role hybridization can play in evolution.
The cell surface expression of group 2 capsular polysaccharides in Escherichia coli: the role of KpsD, RhsA and a multi‐protein complex at the pole of the cell
Authors: McNulty Clodagh , Thompson James , Barrett Brendan , Lord Liz , Andersen Christian , Roberts Ian S.
Published: December 2005

The export of large negatively charged capsular polysaccharides across the outer membrane represents a significant challenge to Gram negative bacteria. In the case of Escherichia coli group 2 capsular polysaccharides, the mechanism of export across the outer membrane was unknown, with no identified candidate outer membrane proteins. In this paper we demonstrate that the KpsD protein, previously believed to be a periplasmic protein, is an outer membrane protein involved in the export of group 2 capsular polysaccharides across the outer membrane. We demonstrate that KpsD and KpsE are located at the poles of the cell and that polysaccharide biosynthesis and export occurs at these polar sites. By in vivo chemical cross‐linking and MALDI‐TOF‐MS analysis we demonstrate the presence of a multi‐protein biosynthetic/export complex in which cytoplasmic proteins involved in polysaccharide biosynthesis could be cross‐linked to proteins involved in export across the inner and outer membranes. In addition, we show that the RhsA protein, of previously unknown function, could be cross‐linked to the complex and that a rhsA mutation reduces K5 biosynthesis suggesting a role for RhsA in coupling biosynthesis and export.
Iron acquisition systems for ferric hydroxamates, haemin and haemoglobin in Listeria monocytogenes
Authors: Jin Bo , Newton Salete M. C. , Shao Yi , Jiang Xiaoxu , Charbit Alain , Klebba Phillip E.
Published: December 2005

Listeria monocytogenes is a Gram‐positive bacterium that causes severe opportunistic infections in humans and animals. We biochemically characterized, for the first time, the iron uptake processes of this facultative intracellular pathogen, and identified the genetic loci encoding two of its membrane iron transporters. Strain EGD‐e used iron complexes of hydroxamates (ferrichrome and ferrichrome A, ferrioxamine B), catecholates (ferric enterobactin, ferric corynebactin) and eukaryotic binding proteins (transferrin, lactoferrin, ferritin, haemoglobin). Quantitative determinations showed 10–100‐fold lower affinity for ferric siderophores (Km ≈ 1–10 nM) than Gram‐negative bacteria, and generally lower uptake rates. Vmax for [59Fe]‐enterobactin (0.15 pMol per 109 cells per minute) was 400‐fold lower than that of Escherichia coli. For [59Fe]‐corynebactin, Vmax was also low (1.2 pMol per 109 cells per minute), but EGD‐e transported [59Fe]‐apoferrichrome similarly to E. coli (Vmax = 24 pMol per 109 cells per minute). L. monocytogenes encodes potential Fur‐regulated iron transporters at 2.031 Mb (the fur‐fhu region), 2.184 Mb (the feo region), 2.27 Mb (the srtB region) and 2.499 Mb (designated hupDGC region). Chromosomal deletions in the fur‐fhu and hupDGC regions diminished iron uptake from ferric hydroxamates and haemin/haemoglobin respectively. In the former locus, deletion of fhuD (lmo1959) or fhuC (lmo1960) strongly reduced [59Fe]‐apoferrichrome uptake. Deletion of hupC (lmo2429) eliminated the uptake of haemin and haemoglobin, and decreased the virulence of L. monocytogenes 50‐fold in mice. Elimination of srtB region genes (Δlmo2185, Δlmo2186, Δlmo2183), both sortase structural genes (ΔsrtB, ΔsrtA, ΔsrtAB), fur and feoB did not impair iron transport. However, deletion of bacterioferritin (Δfri, lmo943; 0.97 Mb) decreased growth and altered iron uptake: Vmax of [59Fe]‐corynebactin transport tripled in this strain, whereas that of [59Fe]‐apoferrichrome decreased 20‐fold.
Interactions between the YycFG and PhoPR two‐component systems in Bacillus subtilis: the PhoR kinase phosphorylates the non‐cognate YycF response regulator upon phosphate limitation
Authors: Howell Alistair , Dubrac Sarah , Noone David , Varughese Kottayil I. , Devine Kevin
Published: December 2005

Two‐component signal transduction systems (TCS) are an important mechanism by which bacteria sense and respond to their environment. Although each two‐component system appears to detect and respond to a specific signal(s), it is now evident that they do not always act independently of each other. In this paper we present data indicating regulatory links between the PhoPR two‐component system that participates in the cellular response to phosphate limitation, and the essential YycFG two‐component system in Bacillus subtilis. We show that the PhoR sensor kinase can activate the YycF response regulator during a phosphate limitation‐induced stationary phase, and that this reaction occurs in the presence of the cognate YycG sensor kinase. Phosphorylation of YycF by PhoR also occurs in vitro, albeit at a reduced level. However, the reciprocal cross‐phosphorylation does not occur. A second level of interaction between PhoPR and YycFG is indicated by the fact that cells depleted for YycFG have a severely deficient PhoPR‐dependent phosphate limitation response and that YycF can bind directly to the promoter of the phoPR operon. YycFG‐depleted cells neither activate expression of phoA and phoPR nor repress expression of the essential tagAB and tagDEF operons upon phosphate limitation. This effect is specific to the PhoPR‐dependent phosphate limitation response because PhoPR‐independent phosphate limitation responses can be initiated in YycFG‐depleted cells.
A major protein component of the Bacillus subtilis biofilm matrix
Authors: Branda Steven S. , Chu Frances , Kearns Daniel B. , Losick Richard , Kolter Roberto
Published: December 2005

Microbes construct structurally complex multicellular communities (biofilms) through production of an extracellular matrix. Here we present evidence from scanning electron microscopy showing that a wild strain of the Gram positive bacterium Bacillus subtilis builds such a matrix. Genetic, biochemical and cytological evidence indicates that the matrix is composed predominantly of a protein component, TasA, and an exopolysaccharide component. The absence of TasA or the exopolysaccharide resulted in a residual matrix, while the absence of both components led to complete failure to form complex multicellular communities. Extracellular complementation experiments revealed that a functional matrix can be assembled even when TasA and the exopolysaccharide are produced by different cells, reinforcing the view that the components contribute to matrix formation in an extracellular manner. Having defined the major components of the biofilm matrix and the control of their synthesis by the global regulator SinR, we present a working model for how B. subtilis switches between nomadic and sedentary lifestyles.
New insights into the pneumococcal fratricide: relationship to clumping and identification of a novel immunity factor
Authors: Håvarstein Leiv Sigve , Martin Bernard , Johnsborg Ola , Granadel Chantal , Claverys Jean‐Pierre
Published: December 2005

In 1971, Tomasz and Zanati discovered that competent pneumococci have a tendency to form aggregates when pelleted by centrifugation and resuspended in 0.01 N HCl by brief vortexing. Interestingly, no clumping was observed with parallel cultures of non‐competent cells treated in the same way. We set out to elucidate the mechanism behind this striking phenomenon, and were able to show that it depends on extracellular DNA that is presumably released by so‐called competence‐induced cell lysis. Competence‐induced cell lysis, which was first described a few years ago, seems to rely on the concerted action of several murein hydrolases. Our results confirmed and extended previous findings by showing that competence‐induced aggregation is abolished in a lytA–lytC double mutant, and absolutely requires CbpD and its N‐terminal CHAP amidase domain. Furthermore, we discovered a novel competence stimulating peptide (CSP)‐induced immunity protein, encoded by the early competence gene comM (spr1762), which protects competent pneumococci against their own lysins. Together, the murein hydrolases and the immunity protein constitutes a CSP‐controlled mechanism that allows competent pneumococci to commit fratricide by killing non‐competent pneumococci sharing the same ecological niche. Through such predatory behaviour, pneumococci can get access to transforming DNA and nutrients, promote the release of virulence factors, and at the same time get rid of competitors.
Functional gastrointestinal disorders and mast cells: implications for therapy
Authors: Barbara G. , Stanghellini V. , De Giorgio R. , Corinaldesi R.
Published: December 2005

Abstract The pathophysiology of functional gastrointestinal disorders is poorly understood. Accepted common mechanisms include psychosocial factors, abnormal gastrointestinal motility and disturbed visceral sensory perception, but the underlying causes remain unclear. Mast cells (MCs) are immunocytes widely distributed throughout the gastrointestinal tract. Several stimuli (e.g. allergens, neuropeptides and stress) lead to MC activation with consequent mediator release (e.g. histamine, tryptase and prostanoids). The MC mediators interact with nerves supplying the gut leading to altered gut physiology and increased sensory perception. The intestinal mucosa of irritable bowel syndrome patients contains on average an increased number of MCs. These cells release an increased amount of mediators in close vicinity to mucosal innervation. The MC activation and their close proximity to nerve fibres is correlated with the severity of perceived abdominal painful sensations. These data provide a strong basis for considering MCs as important participants in visceral hypersensitivity and pain perception in irritable bowel syndrome. Inhibition of MC function may ameliorate irritable bowel symptoms. Novel drugs with an increased potential in the control of MC function (e.g., anti‐IgE antibodies, the intracellular protein tyrosine kinase inhibitor Syk) and mediator release (e.g., second generation antihistamines, proteinase‐activated receptor antagonists) may be useful pharmacological tools for these common disorders.
Neural mechanisms involved in the inhibition of intestinal motility induced by intestinal electrical stimulation in conscious dogs
Authors: Liu S. , Liu J. , Chen J. D. Z
Published: December 2005

Abstract  The effects and mechanisms of intestinal electrical stimulation (IES) with long pulses on intestinal motility were investigated in conscious dogs. Eighteen dogs were equipped with serosal electrodes and an intestinal cannula in the small bowel. The first experiment was designed to study the effect of one‐channel IES on intestinal motility and the extent of this effect. The second experiment was conducted to study the effect of IES on intestinal motility and the involvement of neural pathway. The IES with long pulses significantly inhibited intestinal motility. Intestinal motility of the entire measured segment (40–220 cm distal to the stimulation electrodes) was inhibited by 60–74% with the single‐channel IES with long pulses. Hexamethonium, guanethidine, phentolamine, propranolol partially, but not N‐nitro‐l‐arginine (l‐NNA), ondansetron and naloxone prevented the inhibitory effect of IES on intestinal motility. We conclude that single‐channel IES inhibits intestinal motility within a distance of at least 2 m. This inhibitory effect induced by IES with long pulses is mediated via sympathetic but not nitrergic, serotoninergic 5‐HT3 and opiate pathway.
Control of Penicillium expansum by plant volatile compounds
Authors: Neri F. , Mari M. , Brigati S.
Published: December 2005

Nine plant volatiles were tested for their activity in vitro and in vivo against Penicillium expansum, the cause of blue mould of pear. In vitro spore germination and mycelial growth assay showed a consistent fungicidal activity by trans‐2‐hexenal, carvacrol, trans‐cinnamaldehyde and citral, while hexanal (‐)‐ carvone, p‐anisaldehyde, eugenol and 2‐nonanone exhibited a progressively lower inhibition. trans‐2‐Hexenal was the best inhibitor of conidial germination [MIC (minimum inhibitory concentration) = 24·6 µL L−1; ED50 = 10·2 µL L−1], while carvacrol was the best inhibitor of mycelial growth (MIC = 24·6 µL L−1; ED50 = 9 µL L−1). The four most active compounds in in vitro studies were tested in vivo as fumigants against blue mould on pear cv. Conference. Best control was achieved by trans‐2‐hexenal vapour treatments (12·5 µL L−1) when applied over a 24‐h period, beginning 24 h after inoculation. In contrast, carvacrol (12·5–200 µL L−1), and trans‐cinnamaldehyde (50–400 µL L−1) were ineffective and citral (200 µL L−1) showed only slight effect.
Development of a cliff‐top dune indicated by particle size and geochemical characteristics: Rubjerg Knude, Denmark
Authors: SAYE SAMANTHA E. , PYE KENNETH , CLEMMENSEN LARS B.
Published: December 2005

Particle size and geochemical data have been used to investigate the development of a large cliff‐top dune at Rubjerg Knude, located on the western coast of Jutland, Denmark. Textural parameters and geochemical ratios provided useful indicators of the dune sediment provenance and mode of evolution of the dune. The dune sediments themselves showed no significant spatial particle size trends and reflect a number of processes, including grainfall, wind‐ripple migration and avalanching (grainflow), which formed a high proportion of the deposits on both the stoss and lee sides of the present dune. Fine grainfall sediments, which have accumulated to form a sandplain in the lee of the dune, show fining and improved sorting with distance, and extend more than 2 km downwind of the dune crest. Comparison of the textural and geochemical data from Rubjerg Knude and other locations on the Jutland coast indicates that, although there is a contribution of sand to the dune from local marine sources, the main source of sand to the cliff‐top dune and sand plain sediments has been provided by the wind erosion of the underlying cliffs, which are composed of Weichselian age sandy glaciofluvial and glaciolacustrine deposits. Optically stimulated luminescence dating indicated an apparent age for the sand at the base of 274 ± 14 years. If this date is reliable, it suggests that accumulation of the aeolian sand in this area began within approximately the last 300 years. Map and photographic evidence indicate that the modern high dune only began to form after 1885, apparently associated with an acceleration in the rate of coastal cliff retreat.
Facies distribution of the Lower Cambrian cryptic microbial and epibenthic archaeocyathan‐microbial communities, western Anti‐Atlas, Morocco
Authors: JAVIER ÁLVARO J. , CLAUSEN SÉBASTIEN , ALBANI ABDERRAZZAK EL , CHELLAI EL HASSANE
Published: December 2005

Marine microbial communities recorded in the Moroccan Anti‐Atlas were unaffected across the Neoproterozoic–Cambrian transition. A stromatolite‐dominated consortium was replaced at the beginning of the Atdabanian (ca 20 Myr after the Neoproterozoic–Cambrian boundary) by shelly metazoan and thromboid consortia, which contain the oldest biostratigraphically significant fossils of the Moroccan Cambrian. The associated collapse of microbial mat (stromatolitic) growth appears to coincide with a change from pre‐Atdabanian shallow‐water restricted conditions into Atdabanian deeper, open‐sea conditions. It is postulated that this environmental change led to an episode of improved water circulation over carbonate platform interiors, promoting shelly metazoan immigration into the region. The Tiout/Amouslek lithostratigraphic contact in the early Atdabanian marks the end of an episodically unstable seafloor as suggested by the abundance of slumping and sliding structures, and synsedimentary microfaults and cracks recorded in the underlying Tiout Member. Concurrent with the transition is the occurrence of a network of cryptic fissures and cavities that provided habitats for a coelobiontic chemosynthetic–heterotrophic microbial community composed of stromatolitic crusts, Renalcis–Epiphyton–Girvanella intergrowths, and Kundatia thalli. In the overlying Amouslek Formation, archaeocyathan–thromboid reefs were constrained by substrate stability, water depth and subsidence rate. Four reef geometries are distinguished: (i) patch reefs surrounded by shales, (ii) bioherms in which flank beds intercalate laterally with carbonate and shale inter‐reef sediments, (iii) biostromes or low‐relief structures formed as a result of lateral accretion of patch reefs, and (iv) kalyptrate complexes that nucleated because of a marked tendency for aggregation, and in which patch reefs and bioherms occur stacked together bounded by clay–marl–silt seams.
The BON/CPN gene family represses cell death and promotes cell growth in Arabidopsis
Authors: Yang Shuhua , Yang Huijun , Grisafi Paula , Sanchatjate Siraprapha , Fink Gerald R. , Sun Qi , Hua Jian
Published: December 2005

Copines are calcium‐dependent membrane‐binding proteins that are highly conserved among protozoa, plants, nematodes and mammals. Although they are implicated in membrane trafficking and signal transduction, the functions of these proteins are not well understood. The Arabidopsis copine gene BON1/CPN1 was previously shown to negatively regulate a disease resistance (R) gene SNC1. Here we report that in Arabidopsis, as in other organisms, there is a family of copine genes, BON1, 2 and 3. Using double and triple mutant combinations we show that these three copine genes have overlapping functions essential for the viability of plants. The loss of function of BON1 combined with that of BON2 or BON3 leads to extensive cell death phenotypes resembling the hypersensitive response (HR) in defense responses. The resulting lethality can be suppressed by mutations in PAD4 or EDS1 which are required for R gene signaling and cell death control. Accession‐dependent phenotypes of the mutant combinations suggest that the BON/CPN genes may together repress several R genes other than SNC1. Moreover, the mutant combinations exhibit developmental defects when R‐gene‐mediated defense responses are largely suppressed in pad4 and eds1 mutants. Thus, the copine family in Arabidopsis may have effects in promoting growth and development in addition to repressing cell death, and these two processes might be intricately intertwined.
Choosing between public and private or between hospital and primary care: responsiveness, patient‐centredness and prescribing patterns in outpatient consultations in Bangkok
Authors: Pongsupap Yongyuth , Lerberghe Wim Van
Published: December 2005

Objective  To document differences in provider behaviour between private and public providers in hospital outpatient departments, health centres and clinics in Bangkok, Thailand.

Method  Analysis of the characteristics of 211 taped consultations with simulated patients.

Results  Private hospitals and clinics were significantly more responsive. Private clinics but not private hospitals were also significantly more patient‐centred. All doctors, but particularly those in private hospitals, prescribed unnecessary and potentially harmful technical investigations and drugs. The direct cost to the patient varied between 1.5 (in public health centres) and 12 (in private hospitals) times the minimum daily wage. The combined cost – to the patient and to the state – in public hospitals and health centres exceeded the cost of consultations in private clinics.

Conclusion  Market incentives favour responsiveness and a patient‐centred approach, but not more appropriate therapeutic decisions. Excessive use of pharmaceuticals is observed among public as well as private providers, but is most pronounced in private hospitals. If patients in Bangkok want to maximize responsiveness and degree of patient‐centred care and yet minimize costs and iatrogenesis, they would benefit from avoiding hospitals, both public and private, and, to a lesser extent, specialists. Choosing to use primary facilities, health centres and clinics, particularly when consultations are carried out by general practitioners (GPs), is more beneficial than choosing between public and private providers.
Atopy, eczema and breast milk fatty acids in a high‐risk cohort of children followed from birth to 5 yr
Authors: Oddy Wendy H. , Pal Sebely , Kusel Merci M. H. , Vine Donna , de Klerk Nicholas H. , Hartmann Peter , Holt Patrick G. , Sly Peter D. , Burton Paul R. , Stanley Fiona J. , Landau Louis I.
Published: December 2005

Background: The incidence of atopic diseases such as eczema is increasing in westernized societies. The suggestion that there is a ‘protective’ association between the unique fatty acid composition of breast milk, particularly the omega‐3 (n‐3) and omega‐6 (n‐6) essential polyunsaturated fatty acid content, and the development of atopic disease in children was investigated in a cohort study of 263 infants born into families with a history of allergy (one or both parents had asthma, hayfever, eczema). The objectives of this study were to determine the lipid profile [specifically in relation to long‐chain polyunsaturated fatty acid (LC‐PUFA) composition] in maternal breast milk samples collected at 6 wk and at 6 months following birth, and to investigate the potential role of these fatty acids in modulating the phenotype of children at high genetic risk of developing atopic disease. Method: Breast milk samples were available from 91 atopic mothers at their child's ages of 6 wk and 6 months. These samples were analysed for the fatty acid spectrum. Analysis of variance was used to detect differences between groups of outcomes (no atopy or eczema, non‐atopic eczema, atopy, atopic eczema) at ages 6 months and 5 yr, and a multiple comparisons procedure was conducted to isolate the parameters producing the different results (F‐test, LSD test). For the exposure variables, n‐3 and n‐6 fatty acids are expressed as weight percentage and as a ratio (at both time‐points). Results: The fatty acid profiles of maternal breast milk at 6 wk and 6 months were similar. An increased ratio of n‐6: n‐3 fatty acids in both 6 wk and 6 month milk samples was associated with non‐atopic eczema (p < 0.005) but not atopy alone or atopic eczema. Conclusion: We found milk fatty acids were a significant modulator of non‐atopic eczema but not atopy or atopic eczema in infants at 6 months. In mothers with a history of asthma, hayfever or eczema, their 6‐month‐old infants were more likely to develop non‐atopic eczema if their milk had a higher ratio of n‐6: n‐3 LC‐PUFA.
Association between neopterin in cord blood, urinary neopterin in early childhood and the development of atopic dermatitis, asthma and hay fever
Authors: Horak Elisabeth , Murr Christian , Streif Werner , Schroecksnadel Katharina , Schennach Harald , Fuchs Dietmar
Published: December 2005

It is generally accepted that the increased prevalence of atopic disease is due to a disturbed balance of T‐helper (Th)1/Th2‐type immunity. Upon stimulation by the Th1‐type cytokine interferon (IFN)‐γ, human monocytes/macrophages release large amounts of neopterin. Thus, the determination of neopterin concentrations is an indirect measure of the levels of IFN‐γ and allows us to monitor Th1‐type immune response. We evaluated whether neopterin concentrations in the neonatal cord blood could be a valuable marker predicting atopic disease in early childhood and whether there is a difference in actually determined urinary neopterin concentrations in children with and without atopic disease. Five hundred and five children born during 1997–1999 were enrolled, with cord blood neopterin data available at birth. The International study of asthma and allergies in childhood (ISAAC) questionnaire was used to assess the prevalence of wheezy bronchitis (asthma), atopic dermatitis and allergic rhinitis. Morning urinary samples were collected and urinary neopterin concentration was measured by high‐pressure liquid chromatography. By the average age of 6 yr, the prevalence of atopic disease in the last 12 months was 31%. There was no significant correlation between cord blood and urinary neopterin concentrations at age 6 yr, and between cord blood neopterin and later atopic disease. Urinary neopterin concentrations were significant lower in children with a family history of atopic disease (p = 0.02). In this study, cord blood neopterin concentration was not a predictor for atopic disease in early childhood. Family history of atopic disease was associated with lower urinary neopterin levels at age 6 yr, which might mirror a Th1/Th2 imbalance.
Determination of structure and transcriptional regulation of CYSLTR1 and an association study with asthma and rhinitis
Authors: Zhang Jian , Migita Ohsuke , Koga Minori , Shibasaki Masanao , Arinami Tadao , Noguchi Emiko
Published: December 2005

Pharmacologic studies have revealed that cysteinyl leukotrienes (CYSLTs) act through two receptors, cysteinyl leukotriene receptor 1 (CYSLTR1) and CYSLTR2. CYSLTR1 antagonists are widely used to treat asthma and rhinitis. In this study, we characterized the genomic structure and transcriptional regulation of CYSLTR1 and examined associations between CYSLTR1 polymorphisms and asthma/rhinitis. The experiment of rapid amplification of cDNA end revealed that CYSLTR1 contains three exons and that the entire open reading frame is located in exon 3. Reverse transcriptase‐polymerase chain reaction showed that there were multiple splice variants of CYSLTR1 and that the transcript expression patterns differed from tissues and cell types. The promoter region of CYSLTR1 is from −665 to −30 bp relative to the transcription start site. We identified four polymorphisms (c.‐618‐434T/C, c.‐618‐275C/A, c.‐618‐136G/A, and 927C/T), and transmission disequilibrium tests revealed that none of these polymorphisms was associated with the development of asthma/rhinitis. However, the TCG and CAA haplotypes in the promoter region caused different transcriptional activity. Our findings indicate that CYSLTR1 polymorphisms are not likely to be involved in the development of asthma/rhinitis, but it is possible that these polymorphisms could influence drug responses in individuals with atopic diseases.
Ketotifen induced inhibition of histamine release in a non‐IgE model of middle ear effusion
Authors: Berger Gilead , Berger Rachel , Goldberg Arnon
Published: December 2005

The inhibitory effects of ketotifen, a histamine H1‐receptor antagonist and mast cell stabilizer, were examined on a non‐IgE experimental model of middle ear effusion. Thirty rats were divided into three groups. Group A (n = 9) was subjected to mechanical stimulation of the external auditory canal (EAC); group B (n = 11) was pre‐treated with intraperitoneal administration of 0.2 mg ketotifen, 90 min before mechanical stimulation of the EAC; and group C (n = 10), the control group, was neither exposed to mechanical stimulation nor given ketotifen. Thirty minutes after completion of the experiment, the eardrums were inspected, histamine levels were determined by a fluorometric assay, and the pars flaccida underwent histological assessment. An attic effusion was observed in group A; a similar phenomenon but to a lesser extent was also seen in group B. Statistical analysis confirmed that the mean histamine concentration of the rinsing fluid obtained from group A was significantly higher than that of group C (p = 0.004) or group B (p = 0.008). No significant difference was found between the mean histamine concentration of groups C and group B (p = 0.311). Histological assessment revealed that the thickness of the pars flaccida of group A was considerably greater than that of groups C and B and was characterized by marked edema. Furthermore, the pars flaccida mast cell population was significantly depleted compared with groups C and B. The data indicate that mechanical stimulation of the EAC triggered the pars flaccida mast cells to degranulate in a non‐mediated IgE fashion and that histamine is implicated in most of these histological changes. It is concluded that administration of ketotifen before mechanical stimulation of the EAC had a stabilizing effect and abolished mast cell degranulation, therefore, may be considered as a potential therapeutic agent for the treatment of middle ear disease in the pediatric population.
Domestic allergen and endotoxin exposure and allergic sensitization in Cyprus
Authors: Nicolaou Nicolaos , Yiallouros Panayotis , Pipis Spyros , Ioannou Phivos , Simpson Angela , Custovic Adnan
Published: December 2005

We investigated the relationship between domestic allergen and endotoxin exposure and allergic sensitization among children in Cyprus. We skin prick tested 128 children aged 15–16 yr (random samples of 85 children with self‐reported asthma and 43 healthy controls) and measured their domestic exposure to endotoxin and allergens (mite, cat, and dog). We analyzed the data using multivariate logistic regression (adjusting for gender, area of residence and parental history) and presented the outcomes as odds ratios (OR) and 95% confidence intervals (CI). Among this selected population, 19% of children were sensitized to mite, 15% to cat and 7% to dog. Male gender (OR 2.74, 95% CI 1.18–6.38, p = 0.02), maternal history of allergic disease (OR 3.53, 95% CI 1.13–11.00, p = 0.03), increasing endotoxin (OR 1.58, 95% CI 1.00–2.49, p = 0.05) and residence in the district of Nicosia (OR 2.48, 95% CI 1.01–6.08, p = 0.05) were independent associates of allergic sensitization. Factors associated with mite sensitization were increasing Der p 1 and endotoxin exposure (OR 1.28, 95% CI 1.01–1.62, p = 0.04 and OR 1.76, 95% CI 1.01–3.08, p = 0.05, respectively) and living in an urban area (OR 6.80, 95% CI 1.37–33.67, p = 0.02). Sensitization to domestic pets was associated only with paternal allergic disease (cat: OR 5.68, 95% CI 1.57–23.56, p = 0.02; dog: OR 13.5, 95% CI 1.79–101.73, p = 0.01), but not with pet ownership or specific allergen or endotoxin exposure. In conclusion, mite allergen exposure was associated with sensitization to mite, but there was no association between cat and dog allergen exposure and specific sensitizations. Surprisingly, in this area, increasing endotoxin exposure was associated with an increased risk of sensitization.
Collection of hyperconcentrated platelets with Trima Accel
Authors: Ringwald J. , Duerler T. , Frankow O. , Zimmermann R. , Zingsem J. , Strasser E. , Antoon M. , Eckstein R.
Published: December 2005

Background and Objectives  Lowering the plasma content in single‐donor platelet (PLT) concentrates well below 30% implies the need to collect platelets at very high concentrations. Trima Accel (TA) is validated for collection below 4000 × 103 PLTs/µl. We evaluated its performance at 5000 × 103 PLTs/µl.

Materials and Methods  Twenty blood donors underwent apheresis with TA twice collecting either a hyperconcentrated or a standard single‐donor platelet concentrate with a target platelet concentration of 5000 or 1200 × 103 PLTs/µl, respectively. We analysed the collection efficiency, the collection rate and the quality of the collected by‐plasma.

Results  We collected 20 hyperconcentrated and 20 standard units containing 2·56 ± 0·5 and 3·39 ± 0·4 × 1011 PLTs at a concentration of 4518 ± 978 and 1374 ± 166 × 103 PLTs/µl in 45 ± 8 and 39 ± 6 min resulting in a collection efficiency of 47·5 ± 10·0 and 70·7 ± 7·9% and a collection rate of 5·9 ± 1·4 and 8·8 ± 1·5 × 109 PLTs/min, respectively (all results expressed as mean ± standard deviation). The collected by‐plasma showed a very high grade of cell purity and a satisfactory recovery of the clotting factors.

Conclusion  Although TA is a suitable device for PLT collection at very high concentrations, improvements are desirable to further increase the productivity above its currently validated upper collect concentration limit.
An International Standard for specifying the minimum potency of anti‐D blood‐grouping reagents: evaluation of a candidate preparation in an international collaborative study
Authors: Thorpe S. J. , Fox B. , Heath A. B. , Scott M. , de Haas M. , Kochman S. , Padilla A.
Published: December 2005

Background and Objectives  The aim of this study was to evaluate a lyophilized monoclonal immunoglobulin M (IgM) anti‐D preparation for use as an International Standard to specify a recommended minimum acceptable potency of anti‐D blood‐grouping reagents.

Materials and Methods  The candidate International Standard (99/836) for specifying the minimum potency of anti‐D blood‐grouping reagents was evaluated against a wide range of commercial anti‐D blood‐grouping reagents in an international collaborative study involving 20 laboratories in 13 countries. Laboratories titrated reconstituted 99/836, in parallel with as many commercial anti‐D blood‐grouping reagents as were available to them, in tube tests according to specified haemagglutination methodology for low‐protein (e.g. monoclonal IgM) and high‐protein (e.g. polyclonal) reagents. The ratios of the mean end‐point titres of the reagents to that of 99/836 within each laboratory were calculated.

Results  The ratios of the mean titres of the low‐protein reagents to the mean titre of 99/836 within a laboratory fell between 0·25 and 2 for 43 of the 45 low‐protein anti‐D reagents tested (i.e. the potencies of the low‐protein reagents compared with 99/836 were between a 1 : 4 dilution of 99/836 to twice as potent as 99/836). The ratios of the mean titres of the high‐protein reagents to the mean titre of 99/836 within a laboratory fell within 0·125 and 1 for eight out of the 10 high protein reagents tested.

Conclusions  By international consensus, a 1 : 3 dilution of reconstituted 99/836 was deemed appropriate to define a recommended minimum acceptable potency of low‐protein anti‐D blood‐grouping reagents. A 1 : 8 dilution of reconstituted 99/836 was deemed appropriate to define a recommended minimum acceptable potency of high‐protein anti‐D blood‐grouping reagents. On the basis of the results presented here, 99/836 was established by the World Health Organization as the 1st International Standard for specifying the minimum potency of anti‐D blood‐grouping reagents, in tube tests.
Pathogen inactivation ofLeishmania donovani infantumin plasma and platelet concentrates using riboflavin and ultraviolet light
Authors: Cardo L. J. , Rentas F. J. , Ketchum L. , Salata J. , Harman R. , Melvin W. , Weina P. J. , Mendez J. , Reddy H. , Goodrich R.
Published: December 2005

Background and Objectives Leishmaniais transmitted by the bite of the phlebotomine sandfly or by transfusion of infected blood products. Leishmaniasis currently poses a significant problem in several parts of the world, and is an emerging problem in others. The Mirasol PRT technology is based on the use of riboflavin and ultraviolet light to generate chemical reactions in the nucleic acids of pathogens, which prevents replication and leads to inactivation. The intent of this study was to examine the ability of the Mirasol PRT System to kill theLeishmaniaparasite in human plasma and platelet concentrates.

Materials and Methods In visceral Leishmaniasis, amastigotes are present in the blood and in the reticuloendothelial system within monocytes. For each unit of plasma or platelets treated, isolated mononuclear cells obtained from 100 ml of normal donor whole blood were incubated with 1·0 × 108Leishmania donovani infantumpromastigotes to produce amastigote‐laden macrophages. The infected macrophages were added to 250 ml of human plasma or to 250 ml of platelet concentrates. Infected units were cultured pretreatment in 10‐fold serial dilutions to determine the limits of detection. Thirty millilitres of 500 µmriboflavin was added to each unit, which was then illuminated with 5·9 J/cm2of ultraviolet light (6·24 J/ml). After treatment and after 2 months of frozen storage, plasma units were cultured in 10‐fold serial dilutions. Platelets were cultured on the day of treatment and on day 5 of storage post‐illumination.

Results A 5 log reduction ofLeishmaniawas demonstrated in five of six units of plasma, and a 7 log reduction ofLeishmaniawas demonstrated in one plasma unit. A 5 log reduction ofLeishmaniawas demonstrated in five of six units of platelets, and a 6 log reduction ofLeishmaniawas demonstrated in one unit.

Conclusions There is no donor screen forLeishmaniaand other pathogens constantly emerging in our blood supply. The Mirasol PRT System for Platelets and Plasma is an effective means of killingLeishmaniaand other emerging pathogens in these blood products.
Foraging Ontogeny is not Linked to Delayed Maturation in Squirrel Monkeys (Saimiri sciureus)
Authors: Stone Anita I
Published: December 2005

The acquisition of complex foraging behaviors by young is a proposed cause of a prolonged juvenile phase in many vertebrates, including primates. I compared the foraging behaviors of infant, juvenile and adult squirrel monkeys to determine if significant age‐related differences in foraging behavior and efficiency were present. Infants and juveniles differed from each other in patterns of prey and fruit foraging, but few differences existed between juveniles and adults. Despite differing in the use of foraging substrates, young juveniles (8–12 mo) were as efficient as older juveniles (1–4 yr) and adults at capturing and processing large prey. Young juveniles (<1 yr) were limited in their ability to consume husked palm fruits due to an inability to peel them to obtain pulp. By 1 yr of age, however, foraging behaviors of adults and juveniles were nearly indistinguishable. The absence of meaningful differences between adults and juvenile foraging is not consistent with the hypothesis that the need to develop foraging skills accounts for the pattern of extended juvenility in squirrel monkeys.
Individual Male Calling Pattern and Male Mating Success in the European Treefrog (Hyla arborea): Is there Evidence for Directional or Stabilizing Selection on Male Calling Behaviour?
Authors: Friedl Thomas W.P.
Published: December 2005

In anurans, call properties are commonly classified based on within‐male variability as being either static or dynamic. Numerous playback experiments in the laboratory have indicated that female preferences based on dynamic call properties are usually strongly directional, while female preferences based on static call properties are often stabilizing or weakly directional. However, there are only few studies demonstrating that female preferences for high values of dynamic call properties indeed exert directional selection on male calling behaviour in natural populations. Moreover, field studies investigating whether female preferences for values of static call properties around the mean of the population lead to currently operating stabilizing selection on male calling patterns in natural populations are completely lacking. Here I investigate for two consecutive breeding seasons male calling patterns and male mating success in a population of individually marked European treefrogs (Hyla arborea), a hylid frog with prolonged breeding season and a lek mating system. Individual male calling pattern as analysed in terms of seven temporal and spectral call properties did not differ between males that survived from one breeding season to the next and those not surviving. None of the seven call properties investigated differed significantly between mated and unmated males, indicating that there is no strong directional selection on male calling behaviour in the study population. However, in one study season males that produced calls with a number of pulses around the mean of the population were significantly more likely to obtain matings than males that produced calls with a number of pulses at the low or high end of the distribution. Thus, this study provides preliminary evidence for the operation of stabilizing selection on a static call property (i.e. the number of pulses per call) in a natural population of an anuran amphibian.
Factors affecting health‐related quality of life among patients with lower urinary tract symptoms
Authors: QUEK KIA FATT
Published: December 2005

Background: This study aimed to evaluate the association of factors such as pain, lower urinary tract symptoms (LUTS), anxiety, depression and psychiatric morbidity on health‐related quality of life (HRQoL) of patients with LUTS.

Methods: A total of 227 patients with LUTS were recruited into the study. The patients were assessed on pain, LUTS, anxiety, depression, psychiatric morbidity and health‐related quality of life (HRQoL) using standardized questionnaires.

Results: The study showed that after adjustment, age, pain, anxiety, depression, psychiatric morbidity and LUTS contributed 31%, 13%, 45%, 16% and 48% respectively to the variability of the physical, mental, social, global and overall HRQoL.

Conclusion: Age, pain, psychiatric morbidity, anxiety and depression are associated with HRQoL among patients with LUTS.
Clinical outcomes of laparoscopic adrenalectomy according to tumor size
Authors: HARA ISAO , KAWABATA GAKU , HARA SHOJI , YAMADA YUJI , TANAKA KAZUSHI , FUJISAWA MASATO
Published: December 2005

Objectives: In order to evaluate the indication and usefulness of laparoscopic adrenalectomy, clinical outcomes of laparoscopic adrenalectomy for patients with adrenal tumors were examined. Whether tumor size affects surgical outcome was analysed, along with the long‐term clinical outcome for these patients.

Patients and methods: A total of 63 patients with adrenal tumor underwent laparoscopic adrenalectomy in our institute between 1999 and 2003. A laparoscopic transperitoneal approach was used in all cases. Underlying pathologies comprised Cushing syndrome (n = 12), pheochromocytoma (n = 13), primary aldosteronism (n = 21), non‐functioning adenoma (n = 12) and others (n = 5).

Results: No open conversion was performed. Mean operative duration was 239 min, and mean estimated blood loss was 134 mL. Tumor diameter was significantly smaller for primary aldosteronism than for Cushing syndrome, which in turn was significantly smaller than for adrenocorticotropic hormone‐independent macronodular hyperplasia (AIMAH). No significant differences in surgical outcome and postoperative recovery were noted between large (≥5 cm) and small (<5 cm) tumors. Long‐term clinical outcome was better for patients with pheochromocytoma or primary aldosteronism than for patients with Cushing syndrome.

Conclusions: Laparoscopic adrenalectomy for benign tumor offers excellent surgical outcomes and convalescence. This is true for both small and large tumors.
Bulbocavernosus muscle flap in the repair of complicated vesicovaginal fistula
Authors: XU ZHISHUN , FU QIANG
Published: December 2005

Aim: To investigate the transposition of the bulbocavernosus muscle flap for repairing complicated vesicovaginal fistulas.

Methods: Vesicovaginal fistulas were repaired via combined abdominal and perineal approaches. Through an abdominal approach, the fistula and surrounding scar tissue were excised thoroughly. A perineal incision was made between the orifices of the urethra and the vagina, dissecting until the fistula. The vaginal defect was closed through either the abdominal or the perineal approach depending upon its position. Through the abdominal approach, the bladder defect was closed in two layers with the suture lines vertical to each other. The bulbocavernosus muscle was freed through an incision between the labium majus pudendi and the labium minus pudenda, without damaging the pudendal vascular supply. The bulbocavernosus muscle flap was tunneled beneath the labium minus pudendi, and was sutured in place on the bladder wall over the fistula repair site.

Results: Nine patients with complicated vesicovaginal fistulas were treated using this technique. After surgery, no symptoms of vagina leakage, urinary incontinence, or urethral stricture were reported by any of the patients, and they reported normal sexual function.

Conclusions: Transposition of the bulbocavernosus muscle flap is an excellent technique with low morbidity and high success rate for repairing complicated vesicovaginal fistulas.