bluuebunny/crossref_metadata_2025_split huggingface dataset:
Roles of Amino Acids and Subunits in Determining the Inhibition of Nicotinic Acetylcholine Receptors by Competitive Antagonists
Authors: Dilger James P. , Vidal Ana Maria , Liu Man , Mettewie Claire , Suzuki Takahiro , Pham Anh , Demazumder Deeptankar 
Published: May 2007

Background
Binding sites for agonists and competitive antagonists (nondepolarizing neuromuscular blocking agents) are located at the alpha-delta and alpha-epsilon subunit interfaces of adult nicotinic acetylcholine receptors. Most information about the amino acids that participate in antagonist binding comes from binding studies with (+)-tubocurarine and metocurine. These bind selectively to the alpha-epsilon interface but are differentially sensitive to mutations. To test the generality of this observation, the authors measured current inhibition by five competitive antagonists on wild-type and mutant acetylcholine receptors.

Methods
HEK293 cells were transfected with wild-type or mutant (alphaY198F, epsilonD59A, epsilonD59N, epsilonD173A, epsilonD173N, deltaD180K) mouse muscle acetylcholine receptor complementary DNA. Outside-out patches were excised and perfused with acetylcholine in the absence and presence of antagonist. Concentration-response curves were constructed to determine antagonist IC50. An antagonist-removal protocol was used to determine dissociation and association rates.

Results
Effects of mutations were antagonist specific. alphaY198F decreased the IC50 of (+)-tubocurarine 10-fold, increased the IC50 of vecuronium 5-fold, and had smaller effects on other antagonists. (+)-Tubocurarine was the most sensitive antagonist to epsilonD173 mutations. epsilonD59 mutations had large effects on metocurine and cisatracurium. deltaD180K decreased inhibition by pancuronium, vecuronium, and cisatracurium. Inhibition by these antagonists was increased for receptors containing two delta subunits but no epsilon subunit. Differences in IC50 arose from differences in both dissociation and association rates.

Conclusion
Competitive antagonists exhibited different patterns of sensitivity to mutations. Except for pancuronium, the antagonists were sensitive to mutations at the alpha-epsilon interface. Pancuronium, vecuronium, and cisatracurium were selective for the alpha-delta interface. This suggests the possibility of synergistic inhibition by pairs of antagonists.
Antihyperalgesic Effect of a Recombinant Herpes Virus Encoding Antisense for Calcitonin Gene–related Peptide
Authors: Tzabazis Alexander Z. , Pirc Geanine , Votta-Velis Effrossyni , Wilson Steven P. , Laurito Charles E. , Yeomans David C. 
Published: May 2007

Background
Calcitonin gene-related peptide (CGRP) is contained in and released by small-diameter, nociceptive primary afferent sensory neurons. Upon spinal release, one of the effects of CGRP seems to be to sensitize dorsal horn neurons to subsequent input from nociceptive afferents and, consequently, to induce a behavioral hyperalgesia. Therefore, attenuating evoked release of CGRP from central terminals of nociceptors should have an antihyperalgesic effect.

Methods
The authors applied a recombinant herpes vector, encoding an antisense sequence to the whole CGRP gene, to the dorsal surface of the hind paw of mice to knock down expression of the peptide selectively in primary afferents innervating this tissue.

Results
Herpes virus-based vector encoding an antisense sequence for the whole CGRP clearly reduced CGRP immunoreactivity in the infected spinal dorsal horn levels as well as in cultured dorsal root ganglia neurons. Selective knockdown of CGRP in primary afferents significantly attenuated the thermal, C-fiber hyperalgesia normally observed after topical application of capsaicin. The effect of viral vector-mediated knockdown of CGRP was comparable to the effect of intrathecal application of the CGRP antagonist CGRP8-37, but lasted for 14 weeks after one single application.

Conclusion
Viral vector-mediated knockdown of CGRP in primary afferent neurons provides a promising tool for treatment of chronic pain states as well as for studies investigating the pathophysiology underlying these conditions.
Postoperative Analgesic Effects of Continuous Wound Infiltration with Diclofenac after Elective Cesarean Delivery
Authors: Lavand'homme Patricia M. , Roelants Fabienne , Waterloos Hilde , De Kock Marc F. 
Published: May 2007

Background
Postoperative pain mostly results from sensitization of afferent fibers at injury sites driving central sensitization. Recently, peripheral processes have gained attention as mechanisms of hyperalgesia, and prostaglandins are among highly sensitizing agents. To date, perioperative administration of a single local dose of nonsteroidal antiinflammatory drugs has shown inconclusive efficacy. Rather than a single bolus, the current study evaluates the postoperative analgesic effect of diclofenac continuous intrawound infusion after elective cesarean delivery.

Methods
Ninety-two parturients were randomly allocated to receive a 48-h continuous intrawound infusion with 240 ml containing 300 mg diclofenac, 0.2% ropivacaine, or saline. In the ropivacaine and saline groups, patients also received 75 mg intravenous diclofenac every 12 h for 48 h. Postoperative evaluation included intravenous morphine consumption by patient-controlled analgesia and visual analog pain scores. Punctate mechanical hyperalgesia surrounding the wound and presence of residual pain after 1 and 6 months were also assessed.

Results
Continuous diclofenac infusion significantly reduced postoperative morphine consumption (18 mg; 95% confidence interval, 12.7-22.2) in comparison with saline infusion and systemic diclofenac (38 mg; 95% confidence interval, 28.8-43.7) (P=0.0009) without unique adverse effects. Postoperative analgesia produced by local diclofenac infusion was as effective as local ropivacaine infusion with systemic diclofenac.

Conclusions
After elective cesarean delivery, continuous intrawound infusion of diclofenac demonstrates a greater opioid-sparing effect and better postoperative analgesia than the same dose administered as an intermittent intravenous bolus.
Roles of P2X receptors and Ca2+sensitization in extracellular adenosine triphosphate‐induced hyperresponsiveness in airway smooth muscle
Authors: Oguma T. , Ito S. , Kondo M. , Makino Y. , Shimokata K. , Honjo H. , Kamiya K. , Kume H. 
Published: May 2007

SummaryBackgroundThe release of adenosine triphosphate (ATP) from the airway epithelial cells during the inflammatory process is considered to play an important role in the pathophysiology of asthma and chronic obstructive pulmonary disease.ObjectiveThis study was designed to determine whether extracellular ATP is involved in the bronchial hyperresponsiveness as an interaction between epithelium and smooth muscle in the airways.MethodsWe examined the contractile response to methacholine (MCh) before and after exposure to low concentrations (10 μm) of ATP in isolated, epithelium‐denuded guinea‐pig tracheal smooth muscle by measuring isometric tension. Intracellular Ca2+concentrations ([Ca2+]i) were assessed by fluorescent intensities of fura‐2.ResultsMCh‐induced contractile force was increased with no change in [Ca2+]iafter exposure to 10 μmATP for 15 min. The ability of ATP to enhance the MCh‐induced contraction was markedly attenuated by suramin, a non‐selective P2 receptor inhibitor. Pre‐incubation with ATPγS, a non‐hydrolysable analogue of ATP and α,β‐meATP, a P2X agonist, also enhanced the MCh‐induced contraction. In contrast, uracil triphosphate, a P2Y agonist, did not affect the MCh‐induced contraction. Y‐27632, a Rho‐kinase inhibitor, suppressed the ability of ATP to enhance the MCh‐induced contraction. Moreover, PP1 and PP2, Src tyrosin kinase inhibitors, suppressed the enhancement of MCh‐induced contraction by ATP.ConclusionPre‐treatment with ATP induces hyperresponsiveness to MCh mediated by Ca2+sensitization via the P2X receptor in airway smooth muscle. The present findings suggest the possible involvement of both the Rho‐kinase and Src pathways in the intracellular mechanism of this phenomenon.
The major horse allergen Equ c 1 contains one immunodominant region of T cell epitopes
Authors: Immonen A. , Kinnunen T. , Sirven P. , Taivainen A. , Houitte D. , Peräsaari J. , Närvänen A. , Saarelainen S. , Rytkönen‐Nissinen M. , Maillere B. , Virtanen T. 
Published: May 2007

SummaryBackground
Despite the fact that most significant mammalian respiratory allergens are lipocalin proteins, information on the human T cell reactivity to these allergenic proteins is largely missing.Objective
Knowing the T cell epitopes in allergens is a prerequisite for developing novel preparations for allergen immunotherapy.Methods
Specific T cell lines were generated with recombinant Equ c 1 from the peripheral blood mononuclear cells (PBMCs) of 10 horse‐allergic subjects. For determining T cell epitopes, the lines were stimulated with 16mer synthetic Equ c 1 peptides overlapping by 14 amino acids. The binding capacity of Equ c 1 peptides to human leucocyte antigen class II molecules was determined by the competitive ELISA.Results
The major horse allergen Equ c 1 resembles two other lipocalin allergens, the major cow allergen Bos d 2 and the major dog allergen Can f 1, in that it is weakly stimulatory for the PBMCs of sensitized subjects. Moreover, the T cell epitopes of Equ c 1 are clustered in a few regions along the molecule, as is the case with Bos d 2 and Can f 1. Similar to Bos d 2, Equ c 1 contains one immunodominant epitope region at the carboxy‐terminal end of the molecule. The T cell lines of eight horse‐allergic subjects out of 10 showed strong reactivity to one or both of the two overlapping peptides, p143–158 and p145–160, in this region. The region probably contains two overlapping epitopes.Conclusion
The 18mer peptide p143–160 from the immunodominant region of Equ c 1 is a potential candidate for the peptide‐based immunotherapy of horse‐sensitized subjects.
Immunoreactivity profile of peripheral blood mononuclear cells from patients with ragweed‐induced allergic rhinitis
Authors: Sun J. , Wong B. , Cundall M. , Goncharova S. , Conway M. , Dalrymple A. , Coyle A. J. , Waserman S. , Jordana M. 
Published: May 2007

SummaryBackground
Seasonal rhinitis is manifested by a series of nasal symptoms in response to exposure to seasonal allergens including ragweed pollen. Understanding its immunological mechanisms may help to better manage the disease.Objective
We sought to determine comprehensively ragweed‐induced cytokine and chemokine production by peripheral blood mononuclear cells from normal individuals and patients with seasonal rhinitis sensitized to ragweed pollen, and to assess its regulation by exogenous IL‐10.Methods
Cells were cultured in the presence or absence of a purified ragweed pollen extract with or without exogenous IL‐10. Cytokines and chemokines were measured in the supernatant. Gene expression was evaluated using real‐time quantitative reverse transcription PCR.Results
Ragweed stimulation significantly increased the production of the Th2‐associated cytokines IL‐5, IL‐9 and IL‐13, the chemokines CCL17 and CCL22 and the regulatory cytokine IL‐10 in allergic patients, whereas transforming growth factor‐β (TGF‐β) production was increased only in normal individuals. No difference was detected between groups in the production of the Th1 cytokine IFN‐γ or the Th1‐affiliated chemokines CXCL10 and CXCL11. Exogenous IL‐10 significantly suppressed spontaneous and induced production of both Th1‐ and Th2‐associated cytokines and chemokines.Conclusion
Our work demonstrated that locally manifested allergic rhinitis is underlined by a systemic Th2 immune response specific to allergens. The molecular pathogenesis of allergic rhinitis may be linked to a compromised allergen‐specific immune regulation, e.g., reduced spontaneous and allergen‐induced TGF‐β production in patients compared with healthy controls. Our data also show that IL‐10 inhibits both the effector and directional mechanisms of allergen‐specific immune response, further supporting its potential therapeutic benefit in preventing and treating allergic diseases.
The efficacy of amino acid‐based formulas in relieving the symptoms of cow's milk allergy: a systematic review
Authors: Hill D. J. , Murch S. H. , Rafferty K. , Wallis P. , Green C. J. 
Published: May 2007

SummaryThe aim of this systematic review was to evaluate the efficacy of amino acid‐based formulas (AAF) in patients with cow's milk allergy (CMA). Studies were identified using electronic databases and bibliography searches. Subjects eligible for inclusion were patients of any age with CMA or symptoms suggestive of it. Comparisons of interest were AAF vs. extensively hydrolysed formula (eHF), AAF vs. soy‐based formula (SF) and AAF vs. cow's milk or cow's milk‐based formula. Outcomes of interest were gastrointestinal (GI), dermatological, respiratory and behavioural symptoms as well as growth. A total of 20 studies [three head‐to‐head randomized controlled trials (RCTs), three cross‐over challenge RCTs, seven clinical trials (CTs) and seven case reports (CRs)] were included in the review. In infants with confirmed or suspected CMA, the use of an AAF was shown to be safe and efficacious. Findings from RCT comparisons of AAF with eHF showed that both formulas are equally efficacious at relieving the symptoms of CMA in confirmed or suspected cases. However, infants in specific subgroups (e.g. non‐IgE mediated food‐induced gastro‐enterocolitis‐proctitis syndromes with failure to thrive, severe atopic eczema, or with symptoms during exclusive breastfeeding) were more likely overall to benefit from AAF, as intolerance to eHF may occur. In such cases, symptoms persisting despite eHF feeding usually remit on AAF, and catch‐up growth may be seen. Meta‐analysis of the findings was not possible due to lack of homogenous reporting of outcomes in the original trials. This systematic review shows clinical benefit from use of AAF in both symp̂toms and growth in infants and children with CMA who fail to tolerate eHF. Further studies are required to determine the relative medical or economic value of initial treatment with AAF in infants at high risk of eHF intolerance.
Matrix metalloproteases in vernal keratoconjunctivitis, nasal polyps and allergic asthma
Authors: Leonardi A. , Brun P. , Di Stefano A. , Motterle L. , Abatangelo G. 
Published: May 2007

SummaryBackground
Allergic conditions in different organs share many similarities in their inflammatory response. Vernal keratoconjunctivitis (VKC), asthma and nasal polyps exhibit several similar, but site‐specific mucosal structural changes. The aim of the study was to investigate whether matrix metalloproteases contribute to different tissue remodelling aspects in different organs.Methods
Mucosal biopsies were obtained from conjunctiva of healthy donors, tarsal conjunctiva of vernal patients, bronchi of non‐asthmatic subjects, bronchi of mild stable asthmatic patients, nasal mucosa of non‐allergic donors and nasal polyps of allergic patients. Distribution of metalloprotease‐1, ‐3, ‐9, ‐13, tissue inhibitor of metalloproteases‐1, collagens I and III and the presence of eosinophils and CD4+ cells were evaluated by immunohistochemistry.Results
Collagens were highly diffuse in the giant papillae of VKC and in nasal polyps, and yet less increased in the subepithelium of asthmatic patients. Immunostaining for metalloprotease‐1, ‐3, ‐9 and ‐13 was significantly higher in VKC compared with normal conjunctiva. Metalloprotease‐9 staining was higher in the stroma of polyps vs. normal nasal mucosa, and only metalloprotease‐13 was significantly more expressed in asthmatic vs. non‐asthmatic subjects. Metalloprotease‐9 immunostaining was more intense in vernal compared with other tissues. In all pathological tissues, metalloprotease‐9‐positive staining was in association with eosinophils and CD4+ cells.Conclusions
Expression of metalloproteases may play an important role in inducing the structural changes seen in VKC, nasal polyps and asthma. Tissue remodelling and gelatinase immunoexpression was more dramatic in giant papillae of vernal patients compared with other tissue sites of chronic allergic inflammation.
Human bronchial epithelial cells express an active and inducible biosynthetic pathway for leukotrienes B4 and C4
Authors: Jame A. J. , Lackie P. M. , Cazaly A. M. , Sayers I. , Penrose J. F. , Holgate S. T. , Sampson A. P. 
Published: May 2007

SummaryBackground
Human bronchial epithelial cells synthesize cyclooxygenase and 15‐lipoxygenase products, but the 5‐lipoxygenase (5‐LO) pathway that generates the leukotriene (LT) family of bronchoconstrictor and pro‐inflammatory mediators is thought to be restricted to leucocytes.Objective
We hypothesized that human bronchial epithelial cells (HBECs) express a complete and active 5‐LO pathway for the synthesis of LTB4 and LTC4, either constitutively or after stimulation.Methods
Flow cytometry, RT‐PCR, Western blotting, enzyme immunoassays and reverse‐phase high‐performance liquid chromatography were used to investigate constitutive and stimulated expression of 5‐LO pathway enzymes and the synthesis of LTs B4 and C4 in primary HBECs and in the 16‐HBE 14o− cell line.Results
Constitutive mRNA and protein expression for 5‐LO, 5‐LO‐activating protein (FLAP), LTA4 hydrolase and LTC4 synthase were demonstrated in primary HBECs and in the 16‐HBE 14o− cell line. In 16‐HBE 14o− cells, treatment with calcium ionophore A23187, bradykinin or LPS up‐regulated the expression of these enzymes. The up‐regulation of 5‐LO was blocked by the anti‐inflammatory glucocorticoid dexamethasone. Human bronchial epithelial cells were shown to generate bioactive LTs, with primary HBECs generating 11‐fold more LTC4 and five‐fold more LTB4 than 16‐HBE 14o− cells. LT production was enhanced by ionophore treatment and blocked by the FLAP inhibitor MK‐886.Conclusions
Expression of an active and inducible 5‐LO pathway in HBEC suggests that damaged or inflamed bronchial epithelium may synthesize LTs that contribute directly to bronchoconstriction and leucocytosis in airway inflammation.
Phylogeography of cave pseudoscorpions in southern Australia
Authors: Moulds T. A. , Murphy N. , Adams M. , Reardon T. , Harvey M. S. , Jennings J. , Austin A. D. 
Published: May 2007

AbstractAim To investigate molecular phylogenetic divergence and historical biogeography of the cave‐dwelling pseudoscorpion genusProtochelifer.Location Caves and nearby epigean habitats in southern Australia were sampled from western Victoria, Naracoorte Caves, Flinders Ranges, Kangaroo Island, Nullarbor Plain and south‐west Western Australia.Methods Allozyme electrophoresis (57 individuals) and a 569‐base‐pair section of the mtDNACOIgene (22 individuals) were used to reconstruct phylogenetic relationships among four cave species and three epigean species from 13 locations.Results Phylogenetic reconstruction using the allozyme and mtDNA sequence data revealed a similar topology, showing recent speciation of severalProtocheliferpopulations in caves from Naracoorte to the Nullarbor Plain. Naracoorte Caves contained a single species,Protochelifer naracoortensis, found in four separate caves, while all other cave species appear to be restricted to single caves.Main conclusions At a local scale, as indicated by the four Naracoorte caves, dispersal is thought to occur via micro‐ and mesocaverns, and possibly by phoresy using insect or bat vectors. With current data we are unable to determine if cavernicolous species ofProtocheliferhave arisen from a single cave colonization event followed by phoretic dispersal on bats to other caves, or multiple cave‐invasion events from independent epigean ancestors. Genetic heterogeneity amongProtocheliferpopulations from Nullarbor caves suggest thatP. cavernarum, the only species presently recorded from this region, is likely to constitute a species complex requiring further study to fully resolve its relationships.
Exploring the distribution ofSterocoraxOrtuño, 1990 (Coleoptera, Carabidae) species in the Iberian Peninsula
Authors: Jiménez‐Valverde A. , Ortuño V. M. , Lobo J. M. 
Published: May 2007

AbstractAim The first aim of this paper was to evaluate the distribution of the threeSterocoraxspecies found in the Iberian Peninsula by estimating the main environmental factors that constrain their distributions. The second aim was to explore the potential importance of competitive interactions in limiting their current distributions using predictive distribution models.Location Iberian Peninsula.Methods Species presence data were collected from records in the literature and private and public collections. Ecological niche factor analysis was performed to extract pseudo‐absences (probable absences), which, together with presence data, were modelled using generalized additive models. The models were run twice. Initially we used only environmental variables, and thereafter additional spatial variables were included in order to account for spatially structured factors not accounted for in the environmental variables.Results Highly reliable distribution models were obtained for the three species, with AUC scores (area under the receiver operating characteristics curve) higher than 0.96. The addition of spatial variables to the first model significantly improved the predicted distribution ofCorax (Sterocorax) globosusandCorax (Sterocorax) insidiator, by reducing their potential distribution area. In contrast, the model ofCorax (Sterocorax) galicianuswas not improved by the addition of a spatial term.Main conclusions Generated pseudo‐absences, such as those used in this study, helped to avoid problems of using erroneous data (false absences) in distribution records. Pseudo‐absences greatly improved the models by only selecting absences within the area with the most unfavourable environmental conditions. The importance of spatial variables to bothC. (S.) globosusandC. (S.) insidiatordistributions probably relates to a number of unknown factors, such as unique historical events. The absence of established populations ofC. (S.) globosusnorth of the Ebro Valley appears to be one such historical factor. The distribution ofC. (S.) galicianusonly marginally overlaps with that ofC. (S.) globosus, according to our environmental factor models. As this overlap is restricted it is not likely to be a result of competitive exclusion; rather, their geographical segregation seems to be environmentally mediated. The addition of spatial variables reduced the potential habitat ofC. (S.) insidiator, eliminating some environmentally optimal areas from its distribution. As no environmental barrier seems apparent in this case, competitive interaction withC. (S.) globosusis a plausible hypothesis for its absence in these optimal parts of its range.