bluuebunny/crossref_metadata_2025 huggingface dataset:
Poor correlation between stated and found concentrations of diphenylmethane‐4,4′‐diisocyanate (4,4′‐MDI) in petrolatum patch‐test preparations
Authors: Frick Malin , Zimerson Erik , Karlsson Daniel , Marand Åsa , Skarping Gunnar , Isaksson Marléne , Bruze Magnus
Published: September 2004

Diphenylmethane diisocyanate (MDI) is widely used in its polymeric form in the manufacturing of polyurethane products. Previous reports on MDI‐related contact allergy have shown a pattern, where patients seem to react to their own MDI‐based work material but not to commercial patch‐test preparations, which contain 4,4′‐MDI. Therefore, we performed chemical analyses of 14 commercial test preparations of 4,4′‐MDI obtained from 8 European and 4 American dermatology departments as well as 2 preparations from 2 major European suppliers of patch‐test allergens. A new method for monitoring 4,4′‐MDI in petrolatum preparations was developed and the determination of 4,4′‐MDI as the MDI‐dibutylamine derivative using liquid chromatography‐mass spectrometry was performed. None of the preparations obtained from the dermatology departments contained more than 12% of the concentration stated on the label. In most cases, 4,4′‐MDI content was only a few percentages or less of the concentration stated. 7 of the 14 preparations were analysed before the expiry date. Yet, only 1 of them, a preparation directly obtained from the supplier, came close to the concentration stated on the label. Thus, using these preparations, patients will be tested with a lower concentration than intended, leading to possible false‐negative reactions.
Allergic contact dermatitis from allyl isothiocyanate in a Danish cohort of 259 selected patients
Authors: LerbÆk Anne , Rastogi Suresh Chandra , Menné Torkil
Published: September 2004

Allyl isothiocyanate is present in many plants. Allergic contact dermatitis from allyl isothiocyanate is well known but infrequently reported. The aim of this study was to investigate the prevalence of contact allergy to allyl isothiocyanate in patients with suspected contact dermatitis from vegetables and food. 259 such patients were tested at the Department of Dermatology, Gentofte Hospital, Denmark, from 1994 to 2003. Only 2 patients (0.8%) had a positive reaction (+) to allyl isothiocyanate and 43 patients (16.6%) had a ?+ reaction. One of the patients with a positive reaction provided samples of margarine, salad cream, oil and mayonnaise. These were analysed with high‐performance liquid chromatography, and a moderate concentration of allyl isothiocyanate (2.5 ppm) was detected in the sample of margarine. This patient was a professional sandwich maker presenting with fingertip dermatitis mimicking ‘tulip fingers’ or allergic contact dermatitis from garlic and onions. In conclusion, allergic contact dermatitis from allyl isothiocyanate occurs in only a limited number of cases, despite frequent exposure. The large number of ?+ reactions raises the question as to whether the recommended patch test concentration is too low.
Mystik und Skepsis Gustav Landauers Beitrag zum Literaturverständnis Georg Kaisers
Authors: Krause Frank
Published: September 2004

Eine systematische Untersuchung des Kontakts zwischen Gustav Landauer und Georg Kaiser gilt als Desiderat der Forschung. Als Beitrag zum genaueren Verständnis von Kaisers Begegnung mit Landauer und dessen Schriften sei im folgenden – unter besonderer Berücksichtigung des bislang vernachlässigten Dramas Die Versuchung– nach den Problemen und Perspektiven gefragt, welche die beiden Schriftsteller füreinander bedeutsam werden lassen. Wie sich zeigen wird, findet Landauer Kaisers Werk ethisch und formal überzeugend und möchte es fördern, während Kaiser zur Zeit seines Briefwechsels mit Landauer das offene Grundproblem seiner Literaturauffassung zu lösen sucht: das Mißverhältnis von angestrebter und tatsächlicher Wirkung seiner Stücke. In seiner dramatischen Produktion ab 1916 erprobt Kaiser modifizierte Ansprüche auf soziale Wirkung, und indem Landauer diesen Dramen vorwiegend zustimmt, bestätigt er die Grundtendenz von Kaisers Modifikationen; Landauers bislang wenig bekannte Ablehnung der Versuchung wirkt dieser Tendenz nicht entgegen. Ab 1918 entwickelt Kaiser sein neues Literaturverständnis dann in Aufsätzen, die sich auf Argumente aus Landauers Schriften stützen können. Landauer leistet so einen gewichtigen, für die Expressionismusforschung interessanten Beitrag zur Entwicklung von Kaisers Literaturverständnis.
Poshlust and High Art: A Reading of Nabokov's Aesthetics
Authors: Nyegaard Ole
Published: September 2004

This is a study of the philistine – or the poshlyak– in Nabokov's fiction with focus on Lolita. The concept of poshlost and philistinism holds a special position in his novels and essays. The Russian tradition from which Nabokov drew for his version of the concept is outlined and then compared with other related terms, in order to define it more sharply. The aim is to analyze how Nabokov deploys the concept, not so much as a cultural concept but more as a literary device and finally what this reveals about Nabokov's aesthetics.1
Funktionswandel des Selbstgesprächs Rhetorische Strategien bei Shaftesbury und Johann Jakob Engel
Authors: Finsen Hans Carl
Published: September 2004

Im 18. Jahrhundert wird die Gattung ,Selbstgespräch’ bei Shaftesbury und Johann Jakob Engel zum Gegenstand theoretischer Reflexionen. Bei beiden Autoren bildet das Textproduktionsverfahren den Rahmen der Erörterungen. Die Überlegungen Shaftesburys fügen sich in eine Tradition, die das Selbstgespräch über die aristokratische Konversation auf Cicero zurückführt, und folglich lässt sich bei dem Engländer eine noch weitgehend intakte rhetorische Produktionsstrategie nachweisen. Besonders die in der Forschung wiederholt untersuchte Verquickung von Ethik und Ästhetik ist der rhetorischen Strategie noch prinzipiell verpflichtet. Auch die Reflexionen Engels dokumentieren rhetorische Tradition, vor allem in der Vorliebe für den auch von Shaftesbury bevorzugten leichten Ton, der nun privilegiert im Selbstgespräch erscheint. Gleichzeitig aber verabschiedet Engel die Kunstmäßigkeit als Produktionsprinzip, und so kommt es bei allen Rhetorik‐Reminiszenzen bei diesem Autor dennoch zu einer grundsätzlichen Abkehr von der rhetorischen Textproduktion.
Leconte de Lisle aéronaute: ‘In excelsis’ !
Authors: Mulder Caroline De
Published: September 2004

Loin de se confiner dans une Grèce de marbre, comme l'ont suggéré jusqu’à ce jour les critiques, Leconte de Lisle intègre dans sa poésie des éléments issus des sciences les plus récentes. C'est ce que démontre entres autres ‘In excelsis’, un poème que Leconte de Lisle consacre – quoique de manière voilée –à l'aéronautique. Par ces vers qui n'ont pas retenu l'attention, il s'inscrit dans la lignée des poètes du XIXe siècle qui ont écrit des pièces, souvent très enthousiastes, sur l'aérostat et le voyage aérien. ‘In excelsis’ révèle Leconte de Lisle sous un jour nouveau et permet en outre de cerner les enjeux que présentent dans son œuvre les sciences et les savoirs techniques: ceux‐ci comportent nécessairement, et comme chez d'autres auteurs de la même époque, une dimension prométhéenne et blasphématoire.
AnoXcel: an Anopheles gambiae protein database
Authors: Ribeiro J. M. C. , Topalis P. , Louis C.
Published: September 2004

The proteome of the mosquito Anopheles gambiae was organized on a hyperlinked spreadsheet format containing one protein per row and several pieces of information in each column. The information for each protein ranges from the presence or absence of signal peptide indicative of secretion, presence of transmembrane domains, similarities to several databases, chromosomal location, and relatedness to other An. gambiae proteins, etc. Hosted by AnoBase (http://www.anobase.org/), the whole spreadsheet or segments of it can be downloaded or searched from http://www.anobase.org/AnoBase/Genes/Ano‐Xcel by the scientist dealing with the annotation of proteome subsets such as those deriving from transcriptomes, nucleotide microarrays or high throughput mass spectrometry data.
Apis mellifera ultraspiracle: cDNA sequence and rapid up‐regulation by juvenile hormone
Authors: Barchuk A. R. , Maleszka R. , Simões Z. L. P.
Published: September 2004

Two hormones, 20‐hydroxyecdysone (20E) and juvenile hormone (JH) are key regulators of insect development including the differentiation of the alternative caste phenotypes of social insects. In addition, JH plays a different role in adult honey bees, acting as a ‘behavioural pacemaker’. The functional receptor for 20E is a heterodimer consisting of the ecdysone receptor and ultraspiracle (USP) whereas the identity of the JH receptor remains unknown. We have cloned and sequenced a cDNA encoding Apis mellifera ultraspiracle (AMUSP) and examined its responses to JH. A rapid, but transient up‐regulation of the AMUSP messenger is observed in the fat bodies of both queens and workers. AMusp appears to be a single copy gene that produces two transcripts (∼4 and ∼5 kb) that are differentially expressed in the animal's body. The predicted AMUSP protein shows greater sequence similarity to its orthologues from the vertebrate–crab–tick–locust group than to the dipteran–lepidopteran group. These characteristics and the rapid up‐regulation by JH suggest that some of the USP functions in the honey bee may depend on ligand binding.
Molecular cloning and immunolocalization of a diuretic hormone receptor in rice brown planthopper (Nilaparvata lugens)
Authors: Price D. R. G. , Du J. , Dinsmore A. , Gatehouse J. A.
Published: September 2004

RNA extracted from guts of rice brown planthopper, Nilaparvata lugens, was used to clone cDNA predicted to encode a diuretic hormone receptor (DHR). The DHR, a member of the calcitonin/secretin/corticotropin‐releasing factor family of G‐protein‐coupled receptors, contains seven transmembrane domains and a large N‐terminal extracellular domain potentially involved in hormone binding. The N‐terminal domain was expressed as a recombinant protein, purified and used to raise antibodies. Anti‐DHR IgG bound specifically to Malpighian tubules in immunolocalization experiments using dissected guts, and to a putative DHR polypeptide from N. lugens gut on Western blots. Anti‐DHR IgG delivered orally to insects was not detected in the haemolymph, and showed no binding to gut or tubules, confirming that DHR N‐terminal hormone‐binding domain is not exposed to the gut lumen.
A functional analysis of ACP‐20, an adult‐specific cuticular protein gene from the beetle Tenebrio: role of an intronic sequence in transcriptional activation during the late metamorphic period
Authors: Lemoine A. , Mathelin J. , Braquart‐Varnier C. , Everaerts C. , Delachambre J.
Published: September 2004

A gene encoding the adult cuticular protein ACP‐20 was isolated in Tenebrio. It consists of three exons interspersed by two introns, intron 1 interrupting the signal peptide. To understand the regulatory mechanisms of ACP‐20 expression, ACP‐20 promoter–luciferase reporter gene constructs were transfected into cultured pharate adult wing epidermis. Transfection assays needed the presence of 20‐hydroxyecdysone, confirming that ACP‐20 is up‐regulated by ecdysteroids. Analysis of 5′ deletion constructs revealed that three regions are necessary for high levels of transcription. Interaction experiments between intronic fragments and epidermal nuclear proteins confirmed the importance of intron 1 in ACP‐20 transcriptional control, which results from the combined activity of regulatory cis‐acting elements of the promoter and those of intron 1.
Amino acid residues involved in interaction with tyramine in the Bombyx mori tyramine receptor
Authors: Ohta H. , Utsumi T. , Ozoe Y.
Published: September 2004

To identify amino acid residues interacting with tyramine (TA) in the Bombyx mori TA (BmTA) receptor, several mutant receptors were expressed in HEK‐293 cells and examined for their abilities to bind TA and to attenuate forskolin‐stimulated cAMP production in response to TA. The D134A BmTA receptor showed no specific [3H]TA binding and no TA‐attenuation of cAMP levels. Although the S218A and S222A BmTA receptors showed no specific [3H]TA binding, they still had the ability to mediate the attenuation of cAMP levels in response to the high concentration (100 µm) of TA. The double mutation of Ser218 and Ser222 to Ala, however, led to the loss of TA‐attenuation of cAMP levels. The present study thus confirms that at least three amino acid residues play key roles in interaction with TA in the BmTA receptor.
The dMRP/CG6214 gene of Drosophila is evolutionarily and functionally related to the human multidrug resistance‐associated protein family
Authors: Tarnay J. N. , Szeri F. , Iliás A. , Annilo T. , Sung C. , Le Saux O. , Váradi A. , Dean M. , Boyd C. D. , Robinow S.
Published: September 2004

ATP‐binding cassette (ABC) transporters are involved in the transport of substrates across biological membranes and are essential for many cellular processes. Of the fifty‐six Drosophila ABC transporter genes only white, brown, scarlet, E23 and Atet have been studied in detail. Phylogenetic analyses identify the Drosophila gene dMRP/CG6214 as an orthologue to the human multidrug‐resistance associated proteins MRP1, MRP2, MRP3 and MRP6. To study evolutionarily conserved roles of MRPs we have initiated a characterization of dMRP. In situ hybridization and Northern analysis indicate that dMRP is expressed throughout development and appears to be head enriched in adults. Functional studies indicate that DMRP is capable of transporting a known MRP1 substrate and establishes DMRP as a high capacity ATP‐dependent, vanadate‐sensitive organic anion transporter.
Two amino acid substitutions in acetylcholinesterase associated with pirimicarb and organophosphorous insecticide resistance in the cotton aphid, Aphis gossypii Glover (Homoptera: Aphididae)
Authors: Toda S. , Komazaki S. , Tomita T. , Kono Y.
Published: September 2004

The complete coding sequences of two acetylcholinesterase (AChE) genes, Ace1 (orthologous to Drosophila Ace) and Ace2 (paralogous to Ace), from the cotton aphid (Aphis gossypii) were identified and sequences from carbamate resistant and susceptible strains compared. No change in the amino acid sequences was found in Ace1, while two amino acid substitutions, Ser431Phe and Ala302Ser, were detected between resistant and susceptible strains in Ace2. The position of Ser431Phe corresponds to one of fourteen aromatic residues lining the active site gorge and is located in the acyl pocket. Ala302Ser is located at one of the three residues which form the oxyanion hole in the active site of AChE. The Ser431Phe and Ala302Ser substitutions may play a role in pirimicarb and organophosphate resistance, respectively.
The AeAct‐4 gene is expressed in the developing flight muscles of female Aedes aegypti
Authors: Muñoz D. , Jimenez A. , Marinotti O. , James A. A.
Published: September 2004

Population reduction of mosquitoes is an effective method for controlling dengue fever and malaria transmission. Recent developments in control techniques include proposals to construct transgenic strains of mosquitoes carrying dominant, conditional‐lethal genes under the control of sex‐ and stage‐specific promoters. In order to identify such promoters, subtractive cDNA libraries derived from male and female pupal mRNA of the yellow fever mosquito, Aedes aegypti, were constructed and screened. A cDNA clone, F49, corresponds to a gene expressed specifically in female pupae. Sequence analyses revealed that this gene belongs to the actin gene family, and therefore was designated Aedes Actin‐4 (AeAct‐4). Transcription analyses demonstrated that this gene is expressed predominantly in the indirect flight muscles and, to a lesser extent, the legs of developing female mosquitoes. The promoter of this gene may be a useful tool for developing conditional lethal strains of mosquitoes.
Non‐invasive ventilation in surgical patients in a district general hospital
Authors: Badiger R. , Green M. , Hackwood H. , Palin C. , Shee C. D.
Published: September 2004

We have retrospectively audited the use of non‐invasive ventilation (NIV) in surgical patients. We analysed the case notes of 38 surgical patients who received NIV over a 9‐month period. Twenty‐three patients received NIV following emergency surgery, eight after elective surgery, and seven did not have an operation. Co‐morbidity was common. The commonest reasons for starting NIV were chest infection, acute respiratory distress syndrome and pulmonary oedema. NIV was often only one aspect of treatment in surgical patients with complex medical problems. With intensive support from the critical care outreach team, NIV can be safely delivered on a surgical ward, and may sometimes prevent intensive care unit admission. Use of NIV on the intensive care unit may obviate the need for tracheal intubation in some patients. In very ill surgical patients with a poor prognosis, NIV was frequently used as the ceiling of respiratory support.
Left ventricular mechanical performance in elderly patients after induction of anaesthesia. A comparison of inhalational induction with sevoflurane and intravenous induction with fentanyl and propofol
Authors: Nishikawa K. , Kanaya N. , Kawamata M. , Namiki A.
Published: September 2004

We investigated changes in left ventricular mechanical performance in 40 patients aged > 70 years in whom anaesthesia had been induced with sevoflurane or with fentanyl and propofol. The ratio of ventricular contractility to arterial properties, which reflects left ventricular performance, was estimated from the ratio of ventricular end‐systolic elastance to effective arterial elastance. This ratio decreased after induction in both groups, the magnitude of the decrease being significantly greater in the fentanyl/propofol group than in the sevoflurane group. Decreases in mean arterial pressure after induction of anaesthesia in the two groups were similar, whereas the magnitude of the decrease in heart rate in the sevoflurane group was greater than that in the fentanyl/propofol group. Sevoflurane may therefore be preferable to fentanyl and propofol for induction of anaesthesia in elderly patients because of its lesser effect on left ventricular performance.
Stability of an epidural analgesic admixture containing epinephrine, fentanyl and bupivacaine
Authors: Priston M. J. , Hughes J. M. , Santillo M. , Christie I. W.
Published: September 2004

The addition of epinephrine to solutions containing fentanyl and bupivacaine for epidural infusion has been shown to improve the quality of analgesia. However, this admixture is not available commercially in the United Kingdom. Moreover, stability data applicable to UK practice for this admixture are limited. This study investigated the stability of fentanyl 2 μg.ml−1 plus bupivacaine 1 mg.ml−1 in PVC bags with and without epinephrine 2 μg.ml−1 over a period of 184 days both at room temperature and at 4 °C. All infusions were found to be stable over the study period (> 90% remaining) using stability‐indicating High Performance Liquid Chromatography (HPLC) methods, with no changes in physical appearance or pH (range 4.5–4.2). The infusions were prepared using standard pharmaceutical products, so facilitating the batch preparation of epinephrine, fentanyl and bupivacaine epidural solutions by hospital pharmacy departments.
A pharmacokinetic study of piroxicam in children*
Authors: Dix P. , Prosser D. P. , Streete P.
Published: September 2004

Feldene MeltTM (piroxicam) is commonly used for analgesia following day case surgery. The manufacturer's recommended paediatric dose is 0.4 mg.kg−1 once daily. In children, plasma piroxicam levels of 3–5 μg.ml−1 are associated with effective analgesia. However, in adults a single dose of 20 mg piroxicam (0.4 mg.kg−1 for a 50‐kg adult) produces plasma levels of only 1.5–2.2 μg.ml−1. We therefore studied plasma levels achieved by 0.4 mg.kg−1 or 1.0 mg.kg−1 piroxicam in 22 children aged between 3 and 16 years, undergoing elective orthopaedic surgery, in order to investigate the adequacy of single dosing. The first 12 patients received 0.4 mg.kg−1 Feldene MeltTM pre‐operatively. Following assay of plasma piroxicam levels, a further 10 patients received 1.0 mg.kg−1 Feldene MeltTM. In both groups, five blood samples were taken at 2‐hourly intervals. The mean (95% CI) piroxicam level following 0.4 mg.kg−1 was 2.90 (2.33–3.54) μg.ml−1, compared to 5.87 (4.58–7.16) μg.ml−1 following 1.0 mg.kg−1 (p = 0.0003).
A National Survey of ICU consultant working practices at weekends*
Authors: Thomas E. O.
Published: September 2004

A questionnaire was sent to all Intensive Care Society linkmen to investigate weekend working arrangements on Intensive Care Units (ICU) in the United Kingdom. In all, 87 responses revealed that the average consultant covering ICU at weekends works a 1 in 6 rota, is responsible for 10 beds, works 8–9 h a day and receives two calls at night. Of consultants, 54% cover anaesthesia as well as ICU, 55% work a 48 h or 72 h weekend and only one in five consultants currently have fixed sessional allocation for weekend working. 83% felt that they should not cover anaesthesia as well as ICU and there was no support for consultants to be resident at night. Applying the terms and conditions of the new consultant contract for England to this average consultant would result in 6.6 Programmed Activities for the weekend and 2 days of compensatory rest.
Effect of propofol and isoflurane anaesthesia on the immune response to surgery*
Authors: Inada T. , Yamanouchi Y. , Jomura S. , Sakamoto S. , Takahashi M. , Kambara T. , Shingu K.
Published: September 2004

There are two major subpopulations of peripheral helper T lymphocytes: T helper 1 (Th1) and T helper 2 (Th2) cells. Surgical stress increases the number of Th2 cells, and decreases that of Th1 cells, resulting in a decrease in the Th1/Th2 ratio, and, consequently, in suppressed cell‐mediated immunity. Since anaesthesia can suppress the stress response to surgery, it may inhibit the decrease in the Th1/Th2 ratio. Using flow cytometry, we studied whether propofol anaesthesia (n = 9) or isoflurane anaesthesia (n = 9) had more effect on the decrease in the Th1/Th2 ratio after surgery in patients undergoing craniotomy. The Th1/Th2 ratio decreased significantly after isoflurane anaesthesia (p = 0.011), while it did not change after propofol anaesthesia. The ratio was significantly lower with isoflurane than propofol (p = 0.009). Propofol anaesthesia attenuated the surgical stress‐induced adverse immune response better than isoflurane anaesthesia.
Thyroid storm prior to induction of anaesthesia
Authors: Hirvonen E. A. , Niskanen L. K. , Niskanen M. M.
Published: September 2004

A 53‐ year‐old woman without a previous history of thyroid disease was scheduled for mastectomy. On arrival in the operating theatre unpremedicated she appeared restless and tachycardic. Midazolam and fentanyl was administered intravenously. Concomitantly, sinus tachycardia developed and a flush reaction was observed in the skin of the thoracic region and neck. The blood pressure increased to 265/160 mmHg and the patient lost consciousness and became apnoeic. Unconsciousness and apnoea lasted for approximately 25 min and the operation was postponed. Further investigations revealed an elevated serum free thyroxine level and suppressed serum thyrotropin diagnostic of hyperthyroidism. The serum TSH receptor antibody concentration was elevated, indicating that the patient was suffering from Graves' disease. We present a case of a previously unknown hyperthyroid patient, with breast cancer, presenting as a thyroid crisis on induction of anaesthesia. Although being quite a rare occurrence, unsuspected thyroid disease should be borne in mind when an agitated patient enters the operating theatre.
The Cardiff paediatric laryngoscope blade: a comparison with the Miller size 1 and Macintosh size 2 laryngoscope blades*
Authors: Jones R. M. , Jones P. L. , Gildersleve C. D. , Hall J. E. , Harding L. J. E. , Chawathe M. S.
Published: September 2004

The Cardiff paediatric laryngoscope blade is a single blade that has been designed for use in children from birth to adolescence. This open, randomised, crossover study compared the Cardiff blade with the straight, size 1, Miller laryngoscope blade in 39 infants under 1 years of age and the curved, size 2, Macintosh blade in 39 children aged 1–16 years. The same laryngoscopic view was obtained with the Cardiff and Miller blades in 26 patients; the view was better with the Cardiff blade in seven patients and better with the Miller blade in six (median (IQR [range]) grade of laryngoscopy 1 (1–2 [1–3]) vs. 1 (1–2 [1–3]), respectively; p = 0.405). The Cardiff blade was faster at gaining a view than the Miller blade (mean (SD) time 8.5 (2.9) s vs. 10.2 (3.5) s, respectively; 95% CI for difference −2.8 to −0.4; p = 0.009). The Cardiff and Macintosh blades produced the same view in 32 patients; the view was better with the Cardiff blade in seven patients (median (IQR [range]) grade of laryngoscopy 1 (1–1 [1–3]) vs. 1 (1–2 [1–3]), respectively; p = 0.008). There was no difference in time to gain these views: mean (SD) 8.7 (3.0) s vs. 9.3 (2.7) s, respectively (95% CI for difference −1.58 to 0.40; p = 0.237). The Cardiff paediatric laryngoscope blade compares favourably with these two established laryngoscope blades in children.
Vasopressin and terlipressin: pharmacology and its clinical relevance
Authors: Kam P. C. A. , Williams S. , Yoong F. F. Y.
Published: September 2004

Vasopressin and its analogue, terlipressin, are potent vasopressors that may be useful therapeutic agents in the treatment of cardiac arrest, septic and catecholamine‐resistant shock and oesophageal variceal haemorrhage. The aim of this article is to review the physiology and pharmacology of vasopressin and summarise its efficacy and safety in clinical trials and its subsequent therapeutic use. Recent studies indicate that the use of vasopressin during cardiopulmonary resuscitation may improve the survival of patients with asystolic cardiac arrest. Vasopressin deficiency can contribute to refractory shock states associated with sepsis, cardiogenic shock and cardiac arrest. Low doses of vasopressin and terlipressin can restore vasomotor tone in conditions that are resistant to catecholamines, with preservation of renal blood flow and urine output. They are also useful in reducing bleeding and mortality associated with oesophageal variceal haemorrhage. The long‐term outcome of the use of these drugs is not known.
A high‐resolution radiation hybrid map of porcine chromosome 6
Authors: Cao H. , Robinson J. A. B. , Jiang Z. , Melville J. S. , Golovan S. P. , Jones M. W. , Verrinder Gibbins A. M.
Published: September 2004

A high‐resolution comprehensive map was constructed for porcine chromosome (SSC) 6, where quantitative trait loci (QTL) for reproduction and meat quality traits have been reported to exist. A radiation hybrid (RH) map containing 105 gene‐based markers and 15 microsatellite markers was constructed for this chromosome using a 3000‐rad porcine/hamster RH panel. In total, 40 genes from human chromosome (HSA) 1p36.3‐p22, 29 from HSA16q12‐q24, 17 from HSA18p11.3‐q12 and 19 from HSA19q13.1‐q13.4 were assigned to SSC6. All primers for these gene markers were designed based on porcine gene or EST sequences, and the orthologous status of the gene markers was confirmed by direct sequencing of PCR products amplified from separate Meishan and Large White genomic DNA pools. The RH map spans SSC6 and consists of six linkage groups created by using a LOD score threshold of 4. The boundaries of the conserved segments between SSC6 and HSA1, 16, 18 and 19 were defined more precisely than previously reported. This represents the most comprehensive RH map of SSC6 reported to date. Polymorphisms were detected for 38 of 105 gene‐based markers placed on the RH map and these are being exploited in ongoing chromosome wide scans for QTL and eventual fine mapping of genes associated with prolificacy in a Meishan × Large White multigenerational commercial population.
Fine mapping of the muscular dystrophy (AM) gene on chicken chromosome 2q
Authors: Yoshizawa K. , Inaba K. , Mannen H. , Kikuchi T. , Mizutani M. , Tsuji S.
Published: September 2004

Our previous studies revealed that the genetic locus for chicken muscular dystrophy of abnormal muscle (AM) mapped to chromosome 2q, and that the region showed conserved synteny with human chromosome 8q11‐24.3. In the current study, we mapped the chicken orthologues of genes from human chromosome 8q11‐24 in order to identify the responsible gene. Polymorphisms in the chicken orthologues were identified in the parents of the resource family. Twenty‐three genes and expressed sequence tags (ESTs) were mapped to chicken chromosome 2 by linkage analysis. The detailed comparative map shows a high conservation of synteny between chicken chromosome 2q and human chromosome 8q. The AM locus was mapped between [inositol(myo)‐1(or4)‐monophosphatase 1] (IMPA1) gene and [core‐binding factor, runt domain, α‐subunit 2; translocated to 1; cyclin D‐related] (CBFA2T1) gene. The genes located between IMPA1 and CBFA2T1 are the most likely candidates for chicken muscular dystrophy.
Sequence characterization, polymorphism and chromosomal localizations of the porcine PSME1 and PSME2 genes1
Authors: Wang Y. F. , Yu M. , Te Pas M. F. W. , Yerle M. , Liu B. , Fan B. , Xiong T. A. , Li K.
Published: September 2004

The full‐length cDNA of porcine genes (PSME1 and PSME2) encoding proteasome activators PA28α‐ and β‐subunits were obtained by the rapid amplification of cDNA ends (RACE). The nucleotide sequences and the predicted protein sequences share high sequence identity with their mammalian counterparts. The reverse transcriptase‐polymerase chain reaction (RT‐PCR) revealed that porcine PSME1 and PSME2 genes are expressed in all eight tissues studied (liver, spleen, bladder, small intestine, kidney, heart, skeletal muscle and lung). The full‐length genomic DNA of the porcine PSME1 and PSME2 genes were amplified by PCR. These two genes shared the same structure and were similar in size. A C/T single nucleotide polymorphism in PSME1 intron 8 detected as an SphI PCR‐restriction fragment length polymorphism (PCR–RFLP) shows allele frequency differences between Meishan, Tibetan, Large White, Qingping, and Duroc pigs. The association analysis using two experimental GY selection lines selected for growth rate or leanness suggested that the PSME1 genotype was associated with weaning weight. Analyses of somatic cell hybrid (SCHP) and radiation hybrid (IMpRH) panels revealed that both genes map to SSC7q15.3‐q21 and closely linked to the T‐cell receptor α (TCRA) gene.
Characterization of an X‐chromosome PCR–RFLP marker associated with fat deposition and growth in the pig
Authors: Gaboreanu A.‐M. , Grapes L. , Ramos A. M. , Kim J.‐J. , Rothschild M. F.
Published: September 2004

The X‐chromosome, highly conserved within mammals, has been shown to contain major quantitative trait loci (QTL) for growth and fat deposition in the pig. We have discovered a BamHI polymerase chain reaction restriction fragment length polymorphism (PCR–RFLP) marker that was assigned to the porcine X‐chromosome by two‐point and multi‐point linkage analysis following genotyping of a three‐generation Berkshire by Yorkshire reference family. The marker was positioned 9 cM telomeric to SW2126 and 15.6 cM centromeric to SW1943. Sequence flanking the marker was found to have high similarity to existing database porcine DNA repeat elements. Association analyses of the BamHI marker for growth and meat quality traits in the reference family revealed significant association with marbling (P < 0.03), 10th rib back fat (P < 0.09) and total lipid percentage (P < 0.05), as well as with loin eye area (P < 0.04), average glycolytic potential (P < 0.03) and average lactate content (P < 0.04). Further studies are required to determine the X‐chromosome functional gene affecting fat deposition and growth in the pig.
Characterization and mapping of canine microsatellites isolated from BAC clones harbouring DNA sequences homologous to seven human genes
Authors: Klukowska J. , Szczerbal I. , Wengi‐Piasecka A. , Switonski M. , Schelling C. , Gmür A. , Dolf G.
Published: September 2004

Human primers specific for the genes LEP, HBB, PAX3, ESR2, TPH1, ABCA4 and ATP2A2 were used to identify clones in a canine BAC library. Subcloning of the positive BACs in plasmids, screening with microsatellite motifs and subsequent sequencing allowed for the identification of eight novel microsatellites. The presence of the gene of interest was confirmed by sequencing the polymerase chain reaction (PCR) products amplified in the positive BACs. Fluorescent in situ hybridization (FISH) using the positive BACs as probes allowed for the chromosomal localization of the insert DNAs in two canid species, dog (Canis familiaris) and red fox (Vulpes vulpes). The use of gene‐associated microsatellites may accelerate the identification of candidate genes for phenotypic traits in linkage studies.
Detection of sequence polymorphisms in red junglefowl and White Leghorn ESTs
Authors: Fitzsimmons C. J. , Savolainen P. , Amini B. , Hjälm G. , Lundeberg J. , Andersson L.
Published: September 2004

Over 16 000 high quality expressed sequence tags (ESTs) from red junglefowl (RJ) and White Leghorn (WL) brain and testis cDNA libraries were generated. Here, we have used this resource for detection of single nucleotide polymorphisms (SNPs), and also completed full‐length sequencing of 46 pairs of clones, representing the same gene from both the RJ and WL libraries. From the main set of ESTs, which were assembled using Phrap, 746 putative SNPs were identified, of which 76% were transitions and 24% were transversions. A subset of SNPs was evaluated by sequence analysis of five RJ and five WL birds. Nine of 12 SNPs were verified in this limited sample, suggesting that a majority of the putative polymorphisms documented in this study represent real SNPs. During full‐length sequencing of the 46 RJ/WL clones 100 SNPs were identified, which translated to a frequency of 1.90 SNPs/1000 bp. The number of transitions and transversions were 77% and 23%, respectively, and the proportion of non‐synonymous vs. synonymous SNPs was 20% and 80%, respectively. Four large insertions/deletions were identified between the RJ and WL full‐length sequences, and they appear to represent different splice variants.
Peripheral blood T cell responses to keratin peptides that share sequences with streptococcal M proteins are largely restricted to skin-homing CD8+ T cells
Authors: JOHNSTON A , GUDJONSSON J E , SIGMUNDSDOTTIR H , LOVE T J , VALDIMARSSON H
Published: September 2004

The association of psoriasis with Streptococcus pyogenes throat infections suggests a potential antigenic target for the T cells that are known to infiltrate psoriatic skin. Streptococcal M protein share an extensive sequence homology with the human epidermal keratins. Keratin 17 (K17), while being mostly absent from uninvolved skin, is up-regulated in psoriatic lesions. Consequentially, M-protein-primed T cells may recognize up-regulated keratin epitopes via molecular mimicry. Using in vitro lymphocyte culture and cytokine flow cytometry we demonstrate that HLA-Cw*0602+ psoriasis patients had significant CD8+ T cell interferon (IFN)-γ responses to peptides from the K17 and M6 protein selected on the basis of sequence homology and predicted HLA-Cw*0602 binding. These responses were about 10 times more frequent in the skin-homing cutaneous lymphocyte-associated antigen-expressing (CLA+) subset of CD8+ T cells. CD4+ T cells showed only borderline responses. CLA+ CD8+ T cells from Cw6+ non-psoriatic individuals responded to some M6 peptides but rarely to K17 peptides. Cw6– psoriasis patients showed a response that was intermediate between Cw6+ patients and controls. These findings indicate that psoriatic individuals have CD8+ T cells that recognize keratin self-antigens and that epitopes shared by streptococcal M proteins and human keratins may be targets for the CD8+ T cells that infiltrate psoriatic skin lesions.
Anti-neutrophil cytoplasm antibody IgG subclasses in Wegener's granulomatosis: a possible pathogenic role for the IgG4 subclass
Authors: HOLLAND M , HEWINS P , GOODALL M , ADU D , JEFFERIS R , SAVAGE C O S
Published: September 2004

A characteristic feature of Wegener's granulomatosis is the presence of antineutrophil cytoplasm antibodies (ANCA) to proteinase 3 (PR3). In vitro, ANCA activate neutrophils by co-ligating PR3 and FcγRIIa/IIIb receptors. ANCA are predominantly of the IgG isotype, and IgG1, IgG3 and IgG4 subclasses are particularly represented. To address the pathogenic role of individual ANCA-IgG subclass antibodies, patients’ sera were screened using indirect immunofluorescence, enzyme-linked immunosorbent assay (ELISA) and subclass PR3-ELISA to identify patients with high titres of PR3-ANCA within the IgG1, IgG3 or IgG4 subclasses. Unfractionated ANCA-IgG and subclass fractions were isolated by affinity chromatography and compared for their capacities to stimulate superoxide production by primed human neutrophils. Donor neutrophils were analysed for constitutive and induced FcγRI expression by flow cytometry. The IgG1, IgG3 and IgG4 subclass fractions, isolated from three different ANCA sera, each stimulated superoxide production from neutrophils derived from multiple donors. Subsequently, IgG4 subclass fractions isolated from a further four ANCA positive sera demonstrated varying abilities to stimulate release of superoxide; unrelated to PR3-ANCA titre, neutrophil donor, or neutrophil FcγRI expression. The stimulation of superoxide release by IgG1- and IgG3-ANCA subclass fractions is consistent with the proposed mechanism of co-ligation of PR3 antigen and FcγRIIa/IIIb receptors. However, the demonstration of similar activity for the IgG4-ANCA subclass fractions isolated from some sera was unexpected. This activity was independent of neutrophil donor and expression of FcγRI, suggesting it was capable of activating neutrophils via constitutively expressed FcγRIIa/IIIb or co-ligation of other, unidentified, cell surface molecules.
CD4 T cells producing IFN-γin the lungs of mice challenged with mycobacteria express a CD27-negative phenotype
Authors: LYADOVA I V , OBERDORF S , KAPINA M A , APT A S , SWAIN S L , SAYLES P C
Published: September 2004

Protection against tuberculosis depends upon the generation of CD4+ T cell effectors capable of producing IFN-γ and stimulating macrophage antimycobacterial function. Effector CD4+ T cells are known to express CD44hiCD62Llo surface phenotype. In this paper we demonstrate that a population of CD44hiCD62Llo CD4+ effectors generated in response to Mycobacterium bovis BCG or M. tuberculosis infection in C57BL/6 mice is heterogeneous and consists of CD27hi and CD27lo T cell subsets. These subsets exhibit a similar degree of in vivo proliferation, but differ by the capacity for IFN-γ production. Ex vivo isolated CD27lo T cells express higher amounts of IFN-γ RNA and contain higher frequencies of IFN-γ producers compared to CD27hi subset, as shown by real-time PCR, intracellular staining for IFN-γ and ELISPOT assays. In addition, CD27lo CD4+ T cells uniformly express CD44hiCD62Llo phenotype. We propose that CD27lo CD44hiCD62Llo CD4+ T cells represent highly differentiated effector cells with a high capacity for IFN-γ secretion and antimycobacterial protection at the site of infection.
High prevalence of autoantibodies among Danish centenarians
Authors: ANDERSEN-RANBERG K , HØIER-MADSEN M , WIIK A , JEUNE B , HEGEDÜS L
Published: September 2004

The aim of this study was to investigate the prevalence of organ and nonorgan specific autoantibodies in relation to disability and comorbidity in an unselected population of centenarians. A population-based survey of all persons living in Denmark who celebrated their 100th birthday during the period 1 April 1995 to 31 May 1996, a total of 276 persons, was undertaken. Participants underwent an interview, a physical examination and blood sampling. Organ specific autoantibodies (Tg-ab, TPO-ab, PCA-ab) and nonorgan specific autoantibodies (ANA, IgM RF, IgA RF, MPO-ab, c-ANCA, p-ANCA, oxLDL-ab, IgM ACA, IgG ACA, PR3-ANCA, histone-ab, SSA-ab, SSB-ab, Mit-ab) were measured, and comorbidity and disability (Katz Index of ADL) were registered. In all, 207 (75.0%) of 276 eligible subjects participated, and 148 agreed to blood tests. A large majority (79.3%) had at least one autoantibody detected. Organ specific autoantibodies were present in 32.1% of the centenarians. The high level of autoantibodies did not reflect an equally high level of overt autoimmune disease. While nonorgan specific autoantibodies were equally represented in less-disabled/disabled subjects as well as in subjects with low/high comorbidity, significantly fewer subjects with organ specific autoantibodies were found among less-disabled subjects or subjects with low comorbidity. Autoantibodies (both nonorgan and organ specific) are common in an unselected population of centenarians of today, but do not reflect an equally high level of overt autoimmune disease. Non-organ specific autoantibodies are evenly distributed irrespective of the level of disability or comorbidity, suggesting underlying, undiagnosed pathological processes which may be part of the processes involved in frailty.
IL-10 down-regulates costimulatory molecules onMycobacterium tuberculosis-pulsed macrophages and impairs the lytic activity of CD4 and CD8 CTL in tuberculosis patients
Authors: DE LA BARRERA S , ALEMÁN M , MUSELLA R , SCHIERLOH P , PASQUINELLI V , GARCÍA V , ABBATE E , SASIAIN M DEL C
Published: September 2004

Activation of T cells requires both TCR-specific ligation and costimulation through accessory molecules during T cell priming. IFNγ is a key cytokine responsible for macrophage activation during Mycobacterium tuberculosis (Mtb) infection while IL-10 is associated with suppression of cell mediated immunity in intracellular infection. In this paper we evaluated the role of IFNγ and IL-10 on the function of cytotoxic T cells (CTL) and on the modulation of costimulatory molecules in healthy controls and patients with active tuberculosis (TB). γ-irradiated-Mtb (i-Mtb) induced IL-10 production from CD14+ cells from TB patients. Moreover, CD3+ T cells of patients with advanced disease also produced IL-10 after i-Mtb stimulation. In healthy donors, IL-10 decreased the lytic activity of CD4+ and CD8+ T cells whereas it increased γδ-mediated cytotoxicity. Furthermore, we found that the presence of IL-10 induced a loss of the alternative processing pathways of antigen presentation along with a down-regulation of the expression of costimulatory molecule expression on monocytes and macrophages from healthy individuals. Conversely, neutralization of endogenous IL-10 or addition of IFNγ to either effector or target cells from TB patients induced a strong lytic activity mediated by CD8+ CTL together with an up-regulation of CD54 and CD86 expression on target cells. Moreover, we observed that macrophages from TB patients could use alternative pathways for i-Mtb presentation. Taken together, our results demonstrate that the presence of IL-10 during Mtb infection might contribute to mycobacteria persistence inside host macrophages through a mechanism that involved inhibition of MHC-restricted cytotoxicity against infected macrophages.
111Indium-labelled human gut-derived T cells from healthy subjects with strongin vitroadhesion to MAdCAM-1 show no detectable homing to the gutin vivo
Authors: KELSEN J , AGNHOLT J , FALBORG L , NIELSEN J T , RØMER J L , HOFFMANN H J , DAHLERUP J F
Published: September 2004

Integrin α4β 7 is the principal gut-homing receptor, and it is assumed that expression of this specific integrin directs lymphocytes to the gut in vivo. Adoptive cellular immunotherapy against inflammatory bowel disease (IBD) may depend on the expression of integrin α4β 7 to accomplish local delivery of intravenously injected regulatory T cells in inflamed gut mucosa. The present study aimed to investigate whether in vitro expanded human T cells from the colonic mucosa maintain integrin expression, show in vitro adhesion and retain in vivo gut-homing properties during cultivation. Whole colonic biopsies from healthy subjects were cultured in the presence of interleukin-2 (IL-2) and IL-4. The integrin expression of the cultured T cells was determined by flow cytometry and in vitro adhesion was assessed in a mucosal addressin cell adhesion molecule 1 (MAdCAM-1) adhesion assay. We studied the homing pattern after autologous infusion of 3 × 108 111Indium (111In)-labelled T cells in five healthy subjects using scintigraphic imaging. The cultured CD4+CD45RO+ gut-derived T cells express higher levels of integrin α4β 7 than peripheral blood lymphocytes (PBLs) and show strong adhesion to MAdCAM-1 in vitro, even after 111In-labelling. Scintigraphic imaging, however, showed no gut-homing in vivo. After prolonged transit through the lungs, the T cells migrated preferentially to the spleen, liver and bone marrow. In conclusion, it is feasible to infuse autologous T cells cultured from the gut mucosa, which may be of interest in adoptive immunotherapy. Despite high expression of the gut-homing integrin α4β 7 and adhesion to MAdCAM-1 in vitro, evaluation by 111In-scintigraphy demonstrated no gut-homing in healthy individuals.
Severe respiratory syncytial virus infections and reduced interferon-γ generation in vitro
Authors: SCHAUER U , HOFFJAN S , ROTHOEFT T , BARTZ H , KÖNIG S , FUCHS E , BITTSCHEIDT J , KÖCHLING A , STEPHAN V
Published: September 2004

To study the consequences of the interaction of respiratory syncytial virus (RSV) with dendritic cells in vitro, we established a model of the primary immune response using dendritic cells, autologous naive T cells and the superantigen toxic shock syndrome toxin 1 (TSST 1). About 10% of the naive T cells express the T cell receptor chain Vβ2. These cells were stimulated by TSST 1 and could be analysed by flow cytometry. Cultures infected with RSV produced significantly less interferon-γ compared to uninfected cultures. In a first set of experiments we evaluated whether this culture model using isolated CD4+ CD45RA+ T cells, in fact, reflects the primary immune response. In a prospective study, cells were isolated from 13 children at birth, at 1 year of age and at 4 years of age. RSV reduced interferon-γ production at all the age groups analysed and the results were stable over time within a given individual. In a second set of experiments, we asked whether clinical differences in the course of RSV infection are due to variations in the cellular immune response. At the age of 1 year (5–9 months after the RSV epidemic) dendritic cells and naive T cells were obtained from 27 children with a history of bronchiolitis, from 15 children with a benign course of RSV infection and from 26 controls without RSV infection. The frequency of interferon-γ-producing cells in RSV infected cultures was significantly lower (P < 0.001) in cultures from children with a history of RSV bronchiolitis compared to children with mild RSV infection. Cultures from children without infection displayed a wide range of results. Overall, interferon-γ generation in this group was still lower (P < 0.05) than in the group with mild RSV infection. Because we have ruled out that memory cells play a role in the experiments performed, the most likely explanation for our results is that a high generation of interferon-γ in the primary immune response protects from severe RSV mediated disease.