bluuebunny/crossref_metadata_2025 huggingface dataset:
Neuroprotective Effects of Propofol in Models of Cerebral Ischemia
Authors: Adembri Chiara , Venturi Luna , Tani Alessia , Chiarugi Alberto , Gramigni Elena , Cozzi Andrea , Pancani Tristano , De Gaudio Raffaele A. , Pellegrini-Giampietro Domenico E.
Published: December 2005

Background

Propofol (2,6-diisopropylphenol) has been shown to attenuate neuronal injury in a number of experimental conditions, but studies in models of cerebral ischemia have yielded conflicting results. Moreover, the mechanisms involved in its neuroprotective effects are yet unclear.

Methods

The authors evaluated the neuroprotective effects of propofol in rat organotypic hippocampal slices exposed to oxygen-glucose deprivation, an in vitro model of cerebral ischemia. To investigate its possible mechanism of action, the authors then examined whether propofol could reduce Ca2+-induced rat brain mitochondrial swelling, an index of mitochondrial membrane permeability, as well as the mitochondrial swelling evoked by oxygen-glucose deprivation in CA1 pyramidal cells by transmission electron microscopy. Finally, they evaluated whether propofol could attenuate the infarct size and improve the neurobehavioral outcome in rats subjected to permanent middle cerebral artery occlusion in vivo.

Results

When present in the incubation medium during oxygen-glucose deprivation and the subsequent 24 h recovery period, propofol (10-100 microM) attenuated CA1 injury in hippocampal slices in vitro. Ca2+-induced brain mitochondrial swelling was prevented by 30-100 microM propofol, and so were the ultrastructural mitochondrial changes in CA1 pyramidal cells exposed to oxygen-glucose deprivation. Twenty-four hours after permanent middle cerebral artery occlusion, propofol (100 mg/kg, intraperitoneal) reduced the infarct size by approximately 30% when administered immediately after and up to 30 min after the occlusion. Finally, propofol administered within 30 min after middle cerebral artery occlusion was unable to affect the global neurobehavioral score but significantly preserved spontaneous activity in ischemic rats.

Conclusions

These results show that propofol, at clinically relevant concentrations, is neuroprotective in models of cerebral ischemia in vitro and in vivo and that it may act by preventing the increase in neuronal mitochondrial swelling.
Mechanism of Hepatic Heme Oxygenase-1 Induction by Isoflurane
Authors: Hoetzel Alexander , Leitz Daniel , Schmidt Rene , Tritschler Eva , Bauer Inge , Loop Torsten , Humar Matjaz , Geiger Klaus K. , Pannen Benedikt H. J.
Published: December 2005

Background

The heme oxygenase pathway represents a major cell and organ protective system in the liver. The authors recently showed that isoflurane and sevoflurane up-regulate the inducible isoform heme oxygenase 1 (HO-1). Because the activating cascade remained unclear, it was the aim of this study to identify the underlying mechanism of this effect.

Methods

Rats were anesthetized with pentobarbital intravenously or with isoflurane per inhalation (2.3 vol%). Kupffer cell function was inhibited by dexamethasone or gadolinium chloride. Nitric oxide synthases were inhibited by either N(omega)-nitro-L-arginine methyl ester or S-methyl thiourea. N-acetyl-cysteine served as an antioxidant, and diethyldithiocarbamate served as an inhibitor of cytochrome P450 2E1. Protein kinase C and phospholipase A2 were inhibited by chelerythrine or quinacrine, respectively. HO-1 was analyzed in liver tissue by Northern blot, Western blot, immunostaining, and enzymatic activity assay.

Results

In contrast to pentobarbital, isoflurane induced HO-1 after 4-6 h in hepatocytes in the pericentral region of the liver. The induction was prevented in the presence of dexamethasone (P < 0.05) and gadolinium chloride (P < 0.05). Inhibition of nitric oxide synthases or reactive oxygen intermediates did not affect isoflurane-mediated HO-1 up-regulation. In contrast, chelerythrine (P < 0.05) and quinacrine (P < 0.05) resulted in a blockade of HO-1 induction.

Conclusion

The up-regulation of HO-1 by isoflurane in the liver is restricted to parenchymal cells and depends on Kupffer cell function. The induction is independent of nitric oxide or reactive oxygen species but does involve protein kinase C and phospholipase A2.
Succinylcholine-induced Hyperkalemia in Acquired Pathologic States
Authors: Martyn J A. Jeevendra , Richtsfeld Martina , Warner David O.
Published: December 2005

Lethal hyperkalemic response to succinylcholine continues to be reported, but the molecular mechanisms for the hyperkalemia have not been completely elucidated. In the normal innervated mature muscle, the acetylcholine receptors (AChRs) are located only in the junctional area. In certain pathologic states, including upper or lower motor denervation, chemical denervation by muscle relaxants, drugs, or toxins, immobilization, infection, direct muscle trauma, muscle tumor, or muscle inflammation, and/or burn injury, there is up-regulation (increase) of AChRs spreading throughout the muscle membrane, with the additional expression of two new isoforms of AChRs. The depolarization of these AChRs that are spread throughout the muscle membrane by succinylcholine and its metabolites leads to potassium efflux from the muscle, leading to hyperkalemia. The nicotinic (neuronal) alpha7 acetylcholine receptors, recently described to be expressed in muscle also, can be depolarized not only by acetylcholine and succinylcholine but also by choline, persistently, and possibly play a critical role in the hyperkalemic response to succinylcholine in patients with up-regulated AChRs.
Frequencies of hydrophobic and hydrophilic runs and alternations in proteins of known structure
Authors: Schwartz Russell , King Jonathan
Published: December 2005

Patterns of alternation of hydrophobic and polar residues are a profound aspect of amino acid sequences, but a feature not easily interpreted for soluble proteins. Here we report statistics of hydrophobicity patterns in proteins of known structure in a current protein database as compared with results from earlier, more limited structure sets. Previous studies indicated that long hydrophobic runs, common in membrane proteins, are underrepresented in soluble proteins. Long runs of hydrophobic residues remain significantly underrepresented in soluble proteins, with none longer than 16 residues observed. These long runs most commonly occur as buried α helices, with extended hydrophobic strands less common. Avoiding aggregation of partially folded intermediates during intracellular folding remains a viable explanation for the rarity of long hydrophobic runs in soluble proteins. Comparison between database editions reveals robustness of statistics on aqueous proteins despite an approximately twofold increase in nonredundant sequences. The expanded database does now allow us to explain several deviations of hydrophobicity statistics from models of random sequence in terms of requirements of specific secondary structure elements. Comparison to prior membrane‐bound protein sequences, however, shows significant qualitative changes, with the average hydrophobicity and frequency of long runs of hydrophobic residues noticeably increasing between the database editions. These results suggest that the aqueous proteins of solved structure may represent an essentially complete sample of the universe of aqueous sequences, while the membrane proteins of known structure are not yet representative of the universe of membrane‐associated proteins, even by relatively simple measures of hydrophobic patterns.
Stereoelectronic effects on polyproline conformation
Authors: Horng Jia‐Cherng , Raines Ronald T.
Published: December 2005

The polyproline type II (PPII) helix is a prevalent conformation in both folded and unfolded proteins, and is known to play important roles in a wide variety of biological processes. Polyproline itself can also form a type I (PPI) helix, which has a disparate conformation. Here, we use derivatives of polyproline, (Pro)10, (Hyp)10, (Flp)10, and (flp)10, where Hyp is (2S,4R)‐4‐hydroxyproline, Flp is (2S,4R)‐4‐fluoroproline, and flp is (2S,4S)‐4‐fluoroproline, to probe for a stereoelectronic effect on the conformation of polyproline. Circular dichroism spectral analyses show that 4R electron‐with‐drawing substituents stabilize a PPII helix relative to a PPI helix, even in a solvent that favors the PPI conformation, such as n‐propanol. The stereochemistry at C4 ordains the relative stability of PPI and PPII helices, as (flp)10 forms a mixture of PPI and PPII helices in water and a PPI helix in n‐propanol. The conformational preferences of (Pro)10 are intermediate between those of (Hyp)10/(Flp)10 and (flp)10. Interestingly, PPI helices of (flp)10 exhibit cold denaturation in n‐propanol with a value of Ts near 70°C. Together, these data show that stereoelectronic effects can have a substantial impact on polyproline conformation and provide a rational means to stabilize a PPI or PPII helix.
A new generation of protein display scaffolds for molecular recognition
Authors: Hosse Ralf J. , Rothe Achim , Power Barbara E.
Published: December 2005

Engineered antibodies and their fragments are invaluable tools for a vast range of biotechnological and pharmaceutical applications. However, they are facing increasing competition from a new generation of protein display scaffolds, specifically selected for binding virtually any target. Some of them have already entered clinical trials. Most of these nonimmunoglobulin proteins are involved in natural binding events and have amazingly diverse origins, frameworks, and functions, including even intrinsic enzyme activity. In many respects, they are superior over antibody‐derived affinity molecules and offer an ever‐extending arsenal of tools for, e.g., affinity purification, protein microarray technology, bioimaging, enzyme inhibition, and potential drug delivery. As excellent supporting frameworks for the presentation of polypeptide libraries, they can be subjected to powerful in vitro or in vivo selection and evolution strategies, enabling the isolation of high‐affinity binding reagents. This article reviews the generation of these novel binding reagents, describing validated and advanced alternative scaffolds as well as the most recent nonimmunoglobulin libraries. Characteristics of these protein scaffolds in terms of structural stability, tolerance to multiple substitutions, ease of expression, and subsequent applications as specific targeting molecules are discussed. Furthermore, this review shows the close linkage between these novel protein tools and the constantly developing display, selection, and evolution strategies using phage display, ribosome display, mRNA display, cell surface display, or IVC (in vitro compartmentalization). Here, we predict the important role of these novel binding reagents as a toolkit for biotechnological and biomedical applications.
Protein stabilization by specific binding of guanidinium to a functional arginine‐binding surface on an SH3 domain
Authors: Zarrine‐Afsar Arash , Mittermaier Anthony , Kay Lewis E. , Davidson Alan R.
Published: December 2005

Guanidinium hydrochloride (GuHCl) at low concentrations significantly stabilizes the Fyn SH3 domain. In this work, we have demonstrated that this stabilizing effect is manifested through a dramatic (five‐ to sixfold) decrease in the unfolding rate of the domain with the folding rate being affected minimally. This behavior contrasts to the effect of NaCl, which stabilizes this domain by accelerating the folding rate. These data imply that the stabilizing effect of GuHCl is not predominantly ionic in nature. Through NMR studies, we have identified a specific binding site for guanidinium, and we have determined a dissociation constant of 90mMfor this interaction. The guanidinium‐binding site overlaps with a functionally important arginine‐binding pocket on the domain surface, and we have shown that GuHCl is a specific inhibitor of the peptide‐binding activity of the domain. A different SH3 domain possessing a similar arginine‐binding pocket is also thermodynamically stabilized by GuHCl. These data suggest that many proteins that normally interact with arginine‐containing ligands may also be able to specifically interact with guanidinium. Thus, some caution should be used when using GuHCl as a denaturant in protein folding studies. Since arginine‐mediated interactions are often important in the energetics of protein–protein interactions, our observations could be relevant for the design of small molecule inhibitors of protein–protein interactions.
Evaluation of C‐reactive protein, interleukin‐6, and procalcitonin levels in allogeneic hematopoietic stem cell recipients
Authors: Pihusch Markus , Pihusch Rudolf , Fraunberger Peter , Pihusch Verena , Andreesen Reinhard , Kolb Hans‐Jochem , Holler Ernst
Published: December 2005

Abstract:  Background: Prompt detection of transplant‐related complications (TRC) as infections, acute graft‐versus‐host disease (aGVHD), microangiopathic hemolytic anemia, or veno‐occlusive disease following allogeneic hematopoietic stem cell transplantation (HSCT) is essential.Patients and methods: We conducted a prospective trial on clinical significance of C‐reactive protein (CRP), interleukin‐6 (IL‐6) and procalcitonin (PCT) serum levels in TRC. A total of 350 stem cell recipients were admitted. CRP, IL‐6 and PCT were analyzed prior to conditioning and weekly until 8 wk after HSCT. TRC were recorded weekly throughout the study.Results: CRP (4.4 mg/dL vs. 12.8 mg/dL; P < 0.001), IL‐6 (93 ng/mL vs. 1.138 ng/mL; P < 0.001) and PCT (0.8 ng/dL vs. 5.7 ng/dL; P < 0.001) were increased in infectious complications. Only PCT differentiated between infection and other TRC. Exclusive aGVHD did not increase CRP (4.4 mg/dL vs. 5.7 mg/dL; n.s.), IL‐6 (93 ng/mL vs. 153 ng/mL; n.s.) and PCT (0.8 ng/dL vs. 0.8 ng/dL; n.s.). CRP (6.1 mg/dL vs. 3.1 mg/dL; P < 0.001) and IL‐6 (295 ng/mL vs. 122 ng/mL; P = 0.001) were decreased during steroid therapy, but not PCT (2.3 ng/dL vs. 2.0 ng/dL; n.s.).Conclusion: Our study confirmed CRP, IL‐6 and PCT serum levels as helpful markers for TRC. PCT can differentiate infection from GVHD despite steroid therapy. Further trials are needed focusing on the identification of patients who benefit from early risk stratification.
Association of Epstein–Barr virus with human immunodeficiency virus‐negative peripheral T‐cell lymphomas in Japan
Authors: Hirose Yuko , Masaki Yasufumi , Sawaki Toshioki , Shimoyama Kumiko , Karasawa Hiromi , Kawabata Hiroshi , Fukushima Toshihiro , Ogawa Noriyoshi , Wano Yuji , Umehara Hisanori
Published: December 2005

Abstract:  The association of Epstein–Barr virus (EBV) with human immunodeficiency virus‐negative T‐cell lymphoma was examined in 68 patients using the polymerase chain reaction (PCR) with DNA obtained from formalin‐fixed paraffin‐embedded tissues and an in situ hybridization technique. EBV‐encoded RNA (EBER) was detected in 43 of 68 cases (63%) of peripheral T‐cell lymphoma: in 100% (11 of 11 cases) of NK/T‐cell lymphomas, 70% (14 of 20 cases) of angioimmunoblastic T‐cell lymphomas (AILT) and 49% (18 of 37 cases) of other types of peripheral T‐cell lymphoma. A positive band was also detected at high incidence (36 of 65 cases; 55%) in a PCR analysis using primers to detect the Bam HI‐W fragment of EBV. In the immunohistochemical analysis using a monoclonal antibody to latent membrane protein 1 (LMP‐1) of EBV, one of the EBV‐encoded latent gene products, LMP‐1, was found to be expressed in 13 of 64 cases (20%), but EBNA‐2 was not expressed in all the cases examined (0 of 59 cases; 0%). The 5‐yr survival rate was 28% for peripheral T‐cell lymphomas overall, 0% for NK/T‐cell lymphomas, 38% for AILTs and 28% for other types of peripheral T‐cell lymphoma. The difference in the overall survival rate between NK/T‐cell lymphoma and non‐NK/T‐cell lymphoma was significant (P = 0.0498 by Log‐rank test). Among peripheral T‐cell lymphoma patients overall, the group severely infected with EBV (EBER‐ISH ++) had a lower 5‐yr survival rate (8%) than the group slightly (EBER‐ISH +) or not infected (38%; P = 0.0013).
A unique case of acute promyelocytic leukemia showing monocytic differentiation after ATRA (all‐trans retinoic acid) therapy
Authors: Naeem Mohtashim , Harrison Kathleen , Barton Kevin , Nand Sucha , Alkan Serhan
Published: December 2005

Abstract:  Acute promyelocytic leukemia (PML) is characterized by a reciprocal translocation between chromosomes 15 and 17 resulting in a chimeric PML and retinoic acid receptor alpha (RARA) oncogene. The resultant fusion protein (PML/RARA) is thought to block differentiation of bone marrow cells arrested at the promyelocytic stage. In vitro and in vivo studies have shown that the large majority of APL cells undergo granulocytic maturation after ATRA therapy. We report a unique case of a PML/RARA positive APL patient exhibiting extensive monocytic differentiation after ATRA therapy as documented by morphology, flow cytometry, and FISH studies. We discuss potential dual capability for granulocytic/monocytic differentiation of PML/RARA positive APL cells and implications of monocytic differentiation in the management of APL patients treated with ATRA.
Peroxisome proliferator‐activated receptor‐gamma Pro12Ala polymorphism, Helicobacter pylori infection and non‐cardia gastric carcinoma in Chinese
Authors: LIAO S.‐Y. , ZENG Z.‐R. , LEUNG W. K. , ZHOU S.‐Z. , CHEN B. , SUNG J. J. Y. , HU P.‐J.
Published: December 2005

Background  Peroxisome proliferator‐activated receptor γ inhibits the growth and induces apoptosis of gastric cancer cells. A common polymorphism at codon 12 of this gene (Pro12Ala) has been shown to confer protection against diabetes and colorectal cancer.

Aim  To study the association between peroxisome proliferator‐activated receptor γ gene polymorphism, Helicobacter pylori infection and gastric cancer in Chinese.

Methods  One hundred and four consecutive patients with non‐cardia gastric adenocarcinoma and 104 matched controls were examined. Peroxisome proliferator‐activated receptor γ Pro12Ala polymorphism was analysed by polymerase chain reaction‐restriction fragment length polymorphism.

Results  The frequency of peroxisome proliferator‐activated receptor γ G (Ala12) allele was significantly higher among cancer patients (19.2%) than in control (8.7%; OR 2.5, 95% CI 1.1–5.8). While H. pylori infection was more prevalent in gastric cancer patients (OR 3.0; 95% CI 1.6–5.7), the combination of peroxisome proliferator‐activated receptor γ G allele and H. pylori infection further increased the risk of gastric cancer (OR 12.8, 95% CI 3.2–50.5). The presence of the Ala12 did not increase the risk of gastric cancer in H. pylori‐negative subjects.

Conclusion  Our study suggests the potential association between peroxisome proliferator‐activated receptor γ polymorphism and H. pylori infection in the development of non‐cardia gastric cancer.
Long‐term follow‐up of 13C‐urea breath test results after Helicobacter pylori eradication: frequency and significance of borderline δ13CO2 values
Authors: GISBERT J. P. , OLIVARES D. , JIMENEZ I. , PAJARES J. M.
Published: December 2005

Background  The precise choice of cut‐off point for the 13C‐urea breath test to define whether it is positive or negative represents a controversial issue.

Aim  To quantify the 13C‐urea breath test result for several years following Helicobacter pylori eradication, and to evaluate the frequency and the significance of borderline δ13CO2 values.

Methods  Two‐hundred H. pylori eradicated patients confirmed by 13C‐urea breath test (100 mg of urea, citric acid), and having had repeated this test yearly up to 5 years, were studied. δ13CO2 values between 2 and 5‰ were considered as borderline results.

Results  Eight H. pylori recurrences were observed during 406 patient‐years of follow‐up (1.97% yearly). In two of eight reinfected patients, the reinfection was preceded by a negative δ13CO2 value >2‰. Borderline δ13CO2 values were detected in 4% of the 606 urea breath tests performed, and in 25% when only patients in whom H. pylori recurrence was detected in subsequent urea breath tests were included (P < 0.05). The negative‐predictive value of a post‐treatment δ13CO2 >2‰ for the diagnosis of H. pylori recurrence was 99%.

Conclusions  Positive and negative urea breath test results tend to cluster outside the range between 2‰ and 5‰. Nevertheless, a borderline urea breath test δ value (e.g. very close to the selected cut‐off point) should be interpreted cautiously, and the result should probably be confirmed either by repeating the urea breath test or by other diagnostic methods. On the contrary, a δ13CO2 value <2‰ very confidently confirms H. pylori eradication.
Budesonide foam versus budesonide enema in active ulcerative proctitis and proctosigmoiditis
Authors: GROSS V. , BAR‐MEIR S. , LAVY A. , MICKISCH O. , TULASSAY Z. , PRONAI L. , KUPCINSKAS L. , KIUDELIS G. , POKROTNIEKS J. , KOVÁCS Á. , FASZCZYK M. , RAZBADAUSKAS A. , MARGUS B. , STOLTE M. , MÜLLER R. , GREINWALD R. ,   THE INTERNATIONAL BUDESONIDE FOAM STUDY GROUP
Published: December 2005

Background  Rectal budesonide is an effective treatment of active ulcerative proctitis or proctosigmoiditis.

Aim  To compare the therapeutic efficacy, tolerability and safety, and patient's preference of budesonide foam vs. budesonide enema.

Methods  Patients with active ulcerative proctitis or proctosigmoiditis (clinical activity index >4 and endoscopic index ≥4) were eligible for this double‐blind, double‐dummy, randomized, multicentre study. They received 2 mg/25 mL budesonide foam and placebo enema (n = 265), or 2 mg/100 mL budesonide enema and placebo foam (n = 268) for 4 weeks. Primary endpoint was clinical remission (clinical activity index ≤4) at the final/withdrawal visit (per protocol).

Results  A total of 541 patients were randomized – 533 were evaluable for intention‐to‐treat analysis and 449 for per protocol analysis. Clinical remission rates (per protocol) were 60% for budesonide foam and 66% for budesonide enema (P = 0.02362 for non‐inferiority of foam vs. enema within a predefined non‐inferiority margin of 15%). Both formulations were safe and no drug‐related serious adverse events were observed. Because of better tolerability and easier application most patients preferred foam (84%).

Conclusion  Budesonide foam is as effective as budesonide enema in the treatment of active ulcerative proctitis or proctosigmoiditis. Both budesonide formulations are safe, and most patients prefer foam.
The extent of oesophageal acid exposure overlap among the different gastro‐oesophageal reflux disease groups
Authors: SHAPIRO M. , GREEN C. , FAYBUSH E. M. , ESQUIVEL R. F. , FASS R.
Published: December 2005

Background  Studies have demonstrated that patients with Barrett's oesophagus have the highest oesophageal acid exposure profile, followed by erosive oesophagitis and non‐erosive reflux disease patients, but the exact extent of overlap remains unknown.

Aim  To determine the extent of overlap in oesophageal acid exposure among the different gastro‐oesophageal reflux disease groups.

Methods  A total of 121 patients with gastro‐oesophageal reflux disease underwent an upper endoscopy and were classified as having Barrett's oesophagus, erosive oesophagitis and non‐erosive reflux disease‐all (non‐erosive reflux disease‐positive and functional heartburn). Subsequently, patients underwent pH testing and overlap in oesophageal acid exposure among the different gastro‐oesophageal reflux disease groups was determined.

Results  Of those enrolled, 24 had Barrett's oesophagus, 30 erosive oesophagitis and 28 were non‐erosive reflux disease‐positive. Mean oesophageal acid exposure time was 224.8 ± 35, 134.3 ± 21.9 and 141.3 ± 19.8 min for Barrett's oesophagus, erosive oesophagitis and non‐erosive reflux disease‐positive respectively. Per cent overlap for total, upright and supine time between non‐erosive reflux disease‐positive and erosive oesophagitis was 47.4%, 64.7% and 81.8%, between Barrett's oesophagus and erosive oesophagitis was 47.8%, 40.7% and 24%, and between Barrett's oesophagus and non‐erosive reflux disease‐positive was 31.6%, 37.5% and 20.8% respectively.

Conclusions  Our study demonstrated a high oesophageal acid exposure overlap between patients with non‐erosive reflux disease‐positive and erosive oesophagitis, Barrett's oesophagus and erosive oesophagitis, as well as Barrett's oesophagus and non‐erosive reflux disease‐positive patients.
Histological effects of esomeprazole therapy on the squamous epithelium of the distal oesophagus
Authors: VIETH M. , KULIG M. , LEODOLTER A. , NAUCLÉR E. , JASPERSEN D. , LABENZ J. , MEYER‐SABELLEK W. , LIND T. , WILLICH S. , MALFERTHEINER P. , STOLTE M.
Published: December 2005

Background  Proton pump inhibitor therapy has been reported to reduce proliferative changes of the oesophagus significantly in gastro‐oesophageal reflux disease (GERD).

Aim  To assess the histological effects of esomeprazole treatment on the oesophagus.

Methods  Data were derived from a subgroup of patients participating in the proGERD study, who had either erosive reflux disease (n = 720) or non‐erosive reflux disease (n = 35) and who had biopsy data from two sites [(i) 2 cm above the z‐line and (ii) at the z‐line], obtained at baseline and following treatment with esomeprazole. Proliferative changes of the squamous epithelium were assessed histologically by measuring thickness of the basal cell layer and elongation of the papillae as a percentage of the whole epithelial thickness.

Results  In erosive reflux disease patients, the thickness of the basal cell layer and length of the papillae pretreatment were associated with the severity of oesophagitis (P < 0.05), at both biopsy sites. After esomeprazole treatment, baseline thickness and length of papillae were significantly reduced (P < 0.05) at both biopsy sites in non‐erosive reflux disease and erosive reflux disease patients (particularly those with Los Angeles grades C and D).

Conclusion  This demonstrates a strong correlation between severity of GERD and histological parameters. Esomeprazole therapy resulted in clear reversal of proliferative changes observed prior to treatment in the squamous epithelium at both biopsy locations.
Rabeprazole containing triple therapy to eradicate Helicobacter pylori infection on the Texas‐Mexican border
Authors: CARDENAS V. M. , GRAHAM D. Y. , EL‐ZIMAITY H. M. T. , OPEKUN A. R. , CAMPOS A. , CHAVEZ A. , GUERRERO L.
Published: December 2005

Background  Antibiotic resistance and duration of therapy influence the success of proton‐pump inhibitor‐containing Helicobacter pylori eradication therapy. Clarithromycin resistance is associated with treatment failure.

Aim  To examine the success of a 7‐day rabeprazole‐clarithromycin‐amoxicillin therapy in the study population.

Methods  Adults from Ciudad Juarez with H. pylori infections identified by culture or histology received rabeprazole 20 mg, clarithromycin 0.5 g and amoxicillin 1 g, each b.d. for 7 days. Outcome was assessed by 13C‐urea breath test carried out 4+ weeks after treatment.

Results  A total of 111 patients were enrolled and evaluated by urea breath test; 102 completed the full 7 days therapy. Two deviated from protocol, and five stopped because of adverse events. The cure rate (intention‐to‐treat) was 85% (95% CI: 78–91%); the per‐protocol cure rate was 85% (95% CI: 78–91%). Side‐effects were not serious and only 6.6% of those with adverse events stopped medication. Only three isolates were clarithromycin‐resistant and none was cured. Compliance explained most of the successes.

Conclusions  In the study population a 7‐day rabeprazole triple eradication therapy was both effective and well‐tolerated. Clarithromycin resistance was uncommon. We observed a slightly better outcome but consistent with results from recent large studies in US populations.
Oral administration of pulverized Konjac glucomannan prevents the increase of plasma immunoglobulin E and immunoglobulin G levels induced by the injection of syngeneic keratinocyte extracts in BALB/c mice
Authors: Oomizu S. , Onishi N. , Suzuki H. , Ueda K. , Mochizuki M. , Morimoto K. , Kawamoto S. , Ono K. , Kameyoshi Y. , Hide M.
Published: December 2005

Background
Immunoglobulin (Ig) E plays a key role in the pathogenesis of atopic diseases, such as asthma, atopic dermatitis and allergic rhinitis. Oral administration of pulverized Konjac glucomannan (PKGM) has recently been demonstrated to prevent both plasma IgE elevation and developing dermatitis in NC/Nga mice, a model of atopic dermatitis.

Objective
To clarify the direct effect of PKGM on the increase of plasma IgE, we employed the system of BALB/c mouse that increases IgE level without developing dermatitis in response to continuous injection of the extract of syngeneic keratinocytes, PAM 212 cells (PAM extract).

Methods
Three weeks after the start of feeding with either control or PKGM diet, mice were injected subcutaneously with PAM extract bi‐weekly for 10 weeks. The levels of plasma Igs were measured by enzyme‐linked immunosorbent assay every 2 weeks after the injection. The levels of ɛ germline transcription and the amounts of mRNA for IL‐4, IFN‐γ, GATA‐3 and T bet gene in the spleen were evaluated by real‐time RT‐PCR at the end of the experiment.

Results
On the one hand, PKGM prevented the increase of plasma IgE and IgG (IgG1, IgG2b) induced by PAM extract, and on the other hand, it enhanced the levels of plasma IgG3. However, it did not affect the level of plasma IgM. PKGM also reduced the levels of plasma ovalbumin (OVA)‐specific IgE in OVA‐sensitized mice. Moreover, PKGM attenuated the induction of ɛ germline transcription and expression levels of mRNA for IL‐4, IFN‐γ and GATA‐3 in the spleen of PAM extract‐injected mice. PKGM also attenuated the induction of ɛ germline transcription and mRNA for IFN‐γ and T bet in the spleen of phosphate‐buffered saline‐injected control mice.

Conclusions
These results suggested that oral administration of PKGM prevents the elevation of plasma IgE by suppressing IgE class switching in B cells and/or the commitment development of naive lymphocytes to both T‐helper type 1 (Th1) and Th2.
Anti‐tumor necrosis factor‐α antibody treatment reduces pulmonary inflammation and methacholine hyper‐responsiveness in a murine asthma model induced by house dust
Authors: Kim J. , McKinley L. , Natarajan S. , Bolgos G.L. , Siddiqui J. , Copeland S. , Remick D. G.
Published: December 2005

Background/Aims
Recent studies documented that sensitization and exposure to cockroach allergens significantly increase children's asthma morbidity as well as severity, especially among inner city children. TNF‐α has been postulated to be a critical mediator directly contributing to the bronchopulmonary inflammation and airway hyper‐responsiveness in asthma. This study investigated whether an anti‐TNF‐α antibody would inhibit pulmonary inflammation and methacholine (Mch) hyper‐responsiveness in a mouse model of asthma induced by a house dust extract containing both endotoxin and cockroach allergens.

Methods
A house dust sample was extracted with phosphate‐buffered saline and then used for immunization and two additional pulmonary challenges of BALB/c mice. Mice were treated with an intravenous injection of anti‐TNF‐α antibody or control antibody 1  h before each pulmonary challenge.

Results
In a kinetic study, TNF‐α levels within the bronchoalveolar lavage (BAL) fluid increased quickly peaking at 2 h while BAL levels of IL‐4, IL‐5, and IL‐13 peaked at later time‐points. Mch hyper‐responsiveness was measured 24 h after the last challenge, and mice were killed 24 h later. TNF inhibition resulted in an augmentation of these Th2 cytokines. However, the allergic pulmonary inflammation was significantly reduced by anti‐TNF‐α antibody treatment as demonstrated by a substantial reduction in the number of BAL eosinophils, lymphocytes, macrophages, and neutrophils compared with rat IgG‐treated mice. Mch hyper‐responsiveness was also significantly reduced in anti‐TNF‐α antibody‐treated mice and the pulmonary histology was also significantly improved. Inhibition of TNF significantly reduced eotaxin levels within the lung, suggesting a potential mechanism for the beneficial effects.

These data indicate that anti‐TNF‐α antibody can reduce the inflammation and pathophysiology of asthma in a murine model of asthma induced by a house dust extract.
Mycobacterium tuberculosis infection may protect against allergy in a tuberculosis endemic area
Authors: Obihara C. C. , Kimpen J. L. L. , Gie R. P. , van Lill S. W. , Hoekstra M. O. , Marais B. J. , Schaaf H. S. , Lawrence K. , Potter P. C. , Bateman E. D. , Lombard C. J. , Beyers N.
Published: December 2005

Background
Epidemiological studies have shown an inverse relation of mycobacterial infection and the frequency of allergic diseases and asthma. Recent evidence suggests that allergic inflammation may be inhibited in the presence of chronic and persistent infections, such as that by Mycobacterium tuberculosis (MTB). The relation of tuberculin skin test (TST) size, an accepted marker of MTB infection and the frequency of allergic disease symptoms has not been reported from an area where MTB infection is endemic.

Objective
To investigate the association of TST and allergic disease symptoms, in children living in a tuberculosis (TB) endemic area.

Methods
In this cross‐sectional study, 841 children aged 6–14 years from randomly selected household addresses in two poor communities of Cape Town, South Africa, were investigated with TST and standardized International Study on Asthma and Allergies in Childhood‐based questionnaire on allergic disease symptoms.

Results
Children with positive TST (10 mm) were significantly less likely to have allergic disease symptoms, in particular allergic rhinitis (AR) (adjusted odds ratio 0.43; 95% confidence interval 0.24–0.79) than those with negative TST. This association remained significant after adjusting for possible confounders and correcting for the effect of clustering (>1 child per household address) in the sample. There was a significant inverse linear trend in the relation of TST size in millimetre and the frequency of allergic disease symptoms, in particular AR (P<0.001).

Conclusions
These results of inverse association of strong TST reaction and allergic disease symptoms in children from a TB endemic area are in support of the hypotheses that allergic inflammation may be inhibited by chronic infections, such as MTB.
Stronger nasal responsiveness to cold air in individuals with rhinitis and asthma, compared with rhinitis alone
Authors: Hanes L. S. , Issa E. , Proud D. , Togias A.
Published: December 2005

Background
We have previously proposed that, compared with rhinitis alone, theconstellation of upper and lower airway allergic disease is a manifestation of a more severe form of a syndrome affecting the entire airway. If this is correct, not only the lower, but also the upper airways of patients with asthma and rhinitis should demonstrate more abnormalities compared with patients with rhinitis alone, including higher sensitivity to irritant factors.

Objective
To test the hypothesis that, a previously well‐studied natural nasal stimulus, cold, dry air (CDA), produces a stronger response in subjects with allergic rhinitis (AR) and asthma compared with subjects with AR alone.

Methods
We performed nasal provocation with CDA on 24 individuals with asthma and rhinitis and 17 with rhinitis alone. Prior to and after the challenge, nasal symptoms were recorded using visual analogue scales and nasal lavages were performed to determine histamine and lysozyme levels.

Results
The two groups reacted differently to CDA: after the challenge, patients with rhinitis and asthma reported significantly higher scores for nasal congestion, rhinorrhea and lacrimation. Also in this group, significant increases in histamine and in lysozyme levels in nasal lavage fluids were induced by CDA. In subjects with rhinitis alone, CDA failed to increase histamine or lysozyme levels above baseline. The CDA‐induced change from baseline in histamine was significantly higher in the patients with rhinitis and asthma, compared with the rhinitis‐only group.

Conclusion
Patients with AR and asthma have stronger nasal responsiveness to CDA compared with patients with rhinitis alone. This observation is consistent with the notion that compared with rhinitis alone, the presence of asthma and rhinitis signifies a higher degree of functional abnormality of the entire airway.
Effects of steroid treatment on lung CC chemokines, apoptosis and transepithelial cell clearance during development and resolution of allergic airway inflammation
Authors: Uller L. , Lloyd C. M. , Rydell‐Törmänen K. , Persson C. G. A. , Erjefält J. S.
Published: December 2005

Background
Steroid treatment of allergic eosinophilic airway diseases is considered to attenuate cell recruitment by inhibiting several chemokines and to cause eosinophil clearance through inducement of apoptosis of these cells. However, roles of these mechanisms in the actions of steroids in vivo have not been fully established. Also, as regards clearance of tissue eosinophils other mechanisms than apoptosis may operate in vivo.

Objective
This study explores anti‐inflammatory effects of steroids instituted during either development or resolution of airway allergic inflammation.

Methods
Immunized mice were subjected to week‐long daily allergen challenges (ovalbumin). Steroid treatment was instituted either amidst the challenges or exclusively post‐allergen challenge. CC chemokines, goblet cell hyperplasia, occurrence of eosinophil apoptosis, and airway tissue as well as lumen eosinophilia were examined at different time‐points.

Results
Daily steroids instituted amid the allergen challenges non‐selectively attenuated a range of chemokines, permitted egression of tissue eosinophils into airway lumen to increase, and reduced development of lung tissue eosinophilia. Steroid treatment instituted post‐challenge selectively inhibited the CC‐chemokine regulation upon activation, normal T cell expressed and secrted (RANTES), permitted continued egression of eosinophils into airway lumen, and resolved the tissue eosinophilia. Eosinophil apoptosis rarely occurred at development and resolution of the allergic eosinophilic inflammation whether the animals were steroid treated or not. However, anti‐Fas monoclonal antibodies given to mice with established eosinophilia post‐challenge produced apoptosis of the tissue eosinophils indicating that apoptotic eosinophils, if they occur, are well detectible in vivo.

Conclusion
Airway tissue eosinophils are likely eliminated through egression into airway lumen with little involvement of apoptosis and phagocytosis. Our data further suggest that therapeutic steroids may resolve airway inflammation by permitting clearance of tissue eosinophils through egression and inhibiting RANTES‐dependent cell recruitment to lung tissues.
Cloning, expression and immunological characterization of full‐length timothy grass pollen allergen Phl p 4, a berberine bridge enzyme‐like protein with homology to celery allergen Api g 5
Authors: DeWitt Å. Marknell , Andersson K. , Peltre G. , Lidholm J.
Published: December 2005

Background
Timothy grass pollen is a common cause of respiratory allergy in the temperate regions. The major group 4 allergen, Phl p 4, has previously been purified and studied biochemically and immunologically, but has so far not been produced and characterized as a recombinant protein.

Objective
To clone and characterize timothy grass pollen allergen Phl p 4.

Methods
Full‐length Phl p 4 cDNA was cloned using a PCR‐based strategy including 3′‐and 5′‐RACE. Recombinant Phl p 4 was expressed in Escherichia coli and purified by immobilized metal ion affinity chromatography. Its immunological activity was investigated using experimental ImmunoCAP tests, sera from Phl p 4 sensitized individuals and Phl p 4 reactive polyclonal and monoclonal animal antibodies.

Results
Five full‐length Phl p 4 cDNA clones were analysed. Sequence deviations between the clones were present at nine amino acid positions, and the consensus sequence comprised an open reading frame of 525 amino acids, including a predicted 25‐residue signal peptide. The calculated molecular weight of the deduced mature protein was 55.6 kDa and the isoelectric point 9.9, both consistent with previously observed properties of purified nPhl p 4. Close sequence similarity was found to genomic clones from several other Pooideae grass species and to Bermuda grass pollen allergen BG60. Further, similarity was found to members of the berberine bridge enzyme (BBE) family, including celery allergen Api g 5. Recombinant Phl p 4 bound specific immunoglobulin (Ig)E from 31 of 32 nPhl p 4‐reactive sera, and the IgE binding to rPhl p 4 could be inhibited by nPhl p 4 in a dose‐dependent manner.

Conclusions
Full‐length Phl p 4 cDNA was cloned and showed sequence similarity to members of the BBE family. Recombinant Phl p 4 was produced and shared epitopes with natural Phl p 4.
Absence of systemic immunologic changes during dose build‐up phase and early maintenance period in effective specific sublingual immunotherapy in children
Authors: Dehlink E. , Eiwegger T. , Gerstmayr M. , Kampl E. , Bohle B. , Chen K.‐W. , Vrtala S. , Urbanek R. , Szépfalusi Z.
Published: December 2005

Background
Sublingual immunotherapy (SLIT) has been reported to be a safe treatment for inhalant allergies in children. Yet the immunologic mechanisms resulting in clinical improvement are poorly understood.

Objective
To identify early systemic immunologic changes during the first 8 weeks of clinically effective SLIT to grass pollen, tree pollen or house dust mite in paediatric patients with allergic rhinoconjunctivitis and/or asthma.

Methods
Peripheral blood mononuclear cells and plasma samples of 13 children with reduced symptoms after 1 year of SLIT were obtained before therapy and at 2 and 8 weeks after the initiation of SLIT. Allergen‐specific lymphocyte proliferation assays were performed, and allergen‐induced cytokine production (IL‐2, IL‐4, IL‐10, IFN‐γ, and TGF‐β1) was measured by ELISA and flow cytometry. Allergen‐specific IgE, IgG1, IgG4, and IgA levels in plasma samples were determined in ELISA.

Results
During the first 8 weeks of successful SLIT, allergen‐specific lymphoproliferation (n=13) as well as levels of allergen‐specific intracellular (n=8) and secreted cytokines (n=9) did not change significantly. In addition, no alterations in levels of allergen‐specific Igs (n=7) were observed.

Conclusion
We could not find any early systemic immunologic changes during the first 8 weeks of clinically effective SLIT to inhalant allergens in paediatric patients with allergic rhinoconjunctivitis and/or asthma.
Levan (β‐2, 6‐fructan), a major fraction of fermented soybean mucilage, displays immunostimulating properties via Toll‐like receptor 4 signalling: induction of interleukin‐12 production and suppression of T‐helper type 2 response and immunoglobulin E production
Authors: Xu Q. , Yajima T. , Li W. , Saito K. , Ohshima Y. , Yoshikai Y.
Published: December 2005

Background
Products from the fermentation process of soybeans by Bacillus subtilis (natto) have been shown to possess anti‐tumour and immunomodulatory activities. However, the formulations previously examined were not chemically pure, and this is a major limitation for elucidation of the molecular mechanisms for their activities.

Objective
In order to determine which components in soybean mucilage exert immunostimulatory activities, we examined the activities of their purified forms in vitro and in vivo in mice.

Methods
B. subtilis (natto) and fractions including levan and poly‐γ‐glutamic acid (γ‐PGA) from fermented soybean mucilage were prepared. Levels of cytokine production by mouse macrophage cells after treatment with the fractions were measured by means of ELISA. In vivo effect of levan delivered intragastrically on ovalbumin (OVA)‐specific T‐helper type 2 (Th2) response with IgE production was examined in BALB/c mice that had been immunized intraperitoneally with OVA.

Results
Levan but neither γ‐PGA nor killed B. subtilis (natto) was found to exert strong activity to induce production of IL‐12 p40 and TNF‐α by macrophage cell lines in vitro. Results of experiments using Toll‐like receptor (TLR) 4‐deficient mice and TLR4‐transfected human cell line indicated that TLR4 is involved in pattern recognition of levan. Oral administration of levan in vivo significantly reduced the serum levels of OVA‐specific IgE and Th2 response to OVA in mice immunized with OVA.

Conclusion
Levan is an immunostimulatory moiety in products from the fermentation process of B. subtilis (natto) and may be useful for prevention of allergic disorders with IgE production.
A sensitive monoclonal antibody‐based enzyme‐linked immunosorbent assay to quantify Parietaria judaica major allergens, Par j 1 and Par j 2
Authors: Arilla M. C. , González‐Rioja R. , Ibarrola I. , Mir A. , Monteseirín J. , Conde J. , Martínez A. , Asturias J. A.
Published: December 2005

Background
Parietaria pollen is one of the most important causes of pollinosis in Mediterranean countries. Parietaria judaica pollen extract presents two major allergens, Par j 1 and Par j 2, that belong to the lipid transfer protein family.

Objective
To develop an ELISA for quantification of both major allergens of P. judaica pollen extracts, and to assert correlation of these allergens content with the allergenic activity of extracts.

Methods
Natural Par j 1–Par j 2 allergens were purified by gel filtration, ion exchange, and affinity chromatography and identified by mass spectrometry. Rabbit antisera were obtained using this protein preparation as antigen and used for immunoaffinity purification of nPar j 1–Par j 2. BALB/c mice were immunized with the immunopurified nPar j 1–Par j 2 and after fusion and screening by direct ELISA, 5D4 monoclonal antibody was selected as capture antibody to develop a quantitative two‐site ELISA. Bound proteins were detected by a biotinylated Par j 1–Par j 2‐specific polyclonal antibody.

Results
The optimized ELISA was developed from 25 to 8000 pg/mL of purified Par j 1–Par j 2, and a linear portion of 200–1000 pg/mL. The intraassay and interassay coefficients of variation were lower than 7% and 14% respectively. The assay was very sensitive and specific as it had a detection limit of 25 pg/mL and did not detect reactivity with the same family plants, as Urtica. Par j 1–Par j 2 allergens content was measured in 14 P. judaica and two P. officinalis pollen extracts showing a significant correlation with their allergenic activity measured by enzyme allergosorbent test inhibition.

Conclusions
The results proved the usefulness of the two‐sandwich ELISA for the standardization of Parietaria pollen extracts intended for clinical use, because of its good correlation with allergenic potency.