bluuebunny/crossref_metadata_2025 huggingface dataset:
Heparin-based ELISA reduces background reactivity in virus-like particle-based papillomavirus serology
Authors: Wang Xiaohong , Sapp Martin , Christensen Neil D. , Dillner Joakim
Published: December 2004

The interaction between human papillomavirus (HPV) particles and cell surface heparan sulfate requires intact conformation of the HPV particles. Type-specific HPV serology is currently based on virus-like particles (VLPs) with intact conformation. Presence of incorrectly folded VLPs in VLP preparations is recognized as an important cause of cross-reactivity in HPV serology. Heparin-coated microtitre plates were evaluated for capturing conformationally correct VLPs and improving the type specificity of HPV serology. Hybrid VLPs between HPV16 and HPV11, which had been found to have significant reactivity with children's sera and a batch of HPV18 VLPs that had failed the quality control because of significant reactivity with sera from virginal women, were tested in parallel with heparin ELISA, ordinary ELISA and type-specific mAb capture ELISA. Control sera from children that had detectable reactivity with HPV16/11 hybrid VLPs in ordinary ELISA did not react in heparin-based ELISA, but some hybrid VLPs also had background reactivity in capture ELISAs. Control sera from virginal women that had some reactivity with a poor quality HPV18 VLP preparation in ordinary ELISA had no reactivity in heparin or capture ELISA, suggesting that certain VLP preparations expose cross-reactive epitopes that are not exposed on VLPs with heparin-binding ability. As the sensitivity was similar or only marginally affected by the use of heparin plates, use of heparin-coated plates may improve the type specificity of VLP-based ELISAs and reduce interassay variability attributable to variable quality of different VLP batches.
Frozen section examination of endometrial curettings in the diagnosis of ectopic pregnancy
Authors: Barak Shlomi , Oettinger Moshe , Perri Alex , Cohen Hector I. , Barenboym Rafael , Ophir Ella
Published: December 2004

Background.  The aim of our study was to determine the accuracy and validity of frozen section analysis of endometrial curettings in differentiating between abnormal intrauterine and ectopic pregnancies.

Methods.  A retrospective analysis of the database of the Department of Obstetrics and Gynecology in our institute was performed from January 1998 to September 1999. In 70 women with a suspected ectopic pregnancy a diagnostic curettage was sent for frozen section examination because products of conception could not be identified macroscopically in the curettings. Routine paraffin fixation specimens were also prepared from the endometrial curettings. A frozen section diagnosis was considered correct if it concurred with the final pathologic diagnosis, and incorrect if it differed. The sensitivity, specificity, positive predictive value and negative predictive value of a frozen section in identification of conception products were calculated.

Results.  Of the 70 frozen section studies the diagnosis was correct in 63 (90%), and incorrect in 7 (10%) cases. Of 50 specimens interpreted as negative on frozen sections (no products of conception noted), 6 (12%) were found to contain conception products on final pathologic review. One of the 20 (5%) specimens interpreted as positive by a frozen section failed to demonstrate products of conception on a final pathologic section. The sensitivity of frozen sections in the diagnosis of ectopic pregnancy was 76%, specificity 97.8%; positive predictive value 95%; negative predictive value 88% and accuracy 90%.

Conclusions.  Frozen section examination is a rapid and accurate method for identifying products of conception in endometrial curettings, and may reduce delay in the diagnosis of an ectopic pregnancy and in the institution of therapy.
Multivariate analysis of risk factors for stillbirth in Leon, Mexico
Authors: Romero‐Gutiérrez Gustavo , Martínez‐Ceja Carlos Armando , Abrego‐Olvira Elvira , Ponce‐Ponce de León Ana Lilia
Published: December 2004

Background.  To identify sociodemographic and clinical risk factors for antenatal fetal death in a developing country setting.

Methods.  A case–control study was carried out, including 753 women: 251 had a stillbirth (cases) and 502 had a healthy live birth (controls). Stillbirths were considered as antenatal fetal death after 21 weeks of gestation. Seventeen sociodemographic and clinical risk factors for stillbirth were analyzed.

Statistical analysis.  Student's t‐test or the Mann–Whitney U‐test for continuous data and the χ2‐test or Fisher's exact test for categorical variables were used. Logistic regression analysis was used to find significant predictors for stillbirth. Odds ratios (ORs) with 95% confidence intervals (CIs) were estimated.

Results.  Three risk factors were significant in the logistic regression model: maternal age (OR 1.04, 95% CI 1.00–1.08), antenatal care (OR 0.1, 95% CI 0.0–0.2) and umbilical cord complication (OR 5.8, 95% CI 3.2–10.2). The whole model had a determination coefficient of 0.280, with a χ2‐value of 246.2 (p < 0.001).

Conclusions.  In our setting antenatal care should be considered as the cornerstone in the prevention of stillbirth. With adequate antenatal care both women with advanced maternal age and umbilical cord complication could be identified. This finding could be useful for developing as well as developed country settings to avoid the occurrence of stillbirth.
Effect of density on competition between wild and hatchery‐reared Atlantic salmon for shelter in winter
Authors: Orpwood J. E. , Griffiths S. W. , Armstrong J. D.
Published: December 2004

The effect of varying the density of hatchery‐reared Atlantic salmon Salmo salar on the ability of single wild fish to occupy a shelter is assessed. Although there was strong density‐dependence on sheltering overall, the ability of wild Atlantic salmon parr to occupy a shelter was not affected by the presence of hatchery‐reared fish even when outnumbered by four to one. These findings illustrate a competitive asymmetry for shelter in favour of the wild fish at the densities tested.
Breakdown of the brown trout evolutionary history due to hybridization between native and cultivated fish
Authors: Araguas R. M. , Sanz N. , Pla C. , García‐Marín J. L.
Published: December 2004

Genetic changes in the population structure of brown trout Salmo trutta in the eastern Pyrenees were monitored during the 1990s. Stocking with cultivated exogenous fish has resulted in introgressed populations where those with higher introgression coefficients also have the highest values of heterozygosity. Nevertheless, this increase of local diversity reflects a decrease of genetic differentiation between populations and loss of native alleles. These changes obscure native gene pools and modify evolutionary distinctions among native brown trout populations.
Effect of domestication on ide Leuciscus idus L. reproductive parameters
Authors: Ciesla M. , Wojda R.
Published: December 2004

Ide Leuciscus idus L. is a riverine cyprinid commonly produced as a subsidiary species in polyculture with common carp in Poland. In many cases production of restocking material is based on farm‐reared broodstock. The aim of the paper is to study possible domestication effects on female reproductive parameters in ide following long‐term pond cultivation. Three groups of fish were analysed: group (1) consisted of fish cultivated in carp ponds for three generations; group (2) consisted of fish caught as wild one‐year‐olds in rivers and kept in ponds until till sexual maturation (first pond‐reared generation) and group (3) consisted of wild ide females caught at spawning sites just before natural spawning. All fish originated from the Vistula River in central Poland. Fish were stimulated with Hungarian synthetic hormone analogue (Ovopel). Body length and mass, individual fecundity and egg size were measured during spawning, and egg survival was assessed at the eyed ova stage.
Reared perch v. wild perch – first results about organoleptical and technological quality
Authors: Mairesse G. , Thomas M. , Gardeur J.‐N. , Brun‐Bellut J.
Published: December 2004

The Eurasian perch, Perca fluviatilis L., has good potential as a candidate for inland aquaculture diversification in Europe. To aid the development of this new product, we define a quality reference based on attributes of wild perch, and compare the quality of perch reared in three different culture systems: (1) extensive (pond), (2) semi‐extensive (tank) and (3) intensive (re‐circulation system). In order to define a precise quality reference, it is necessary to characterize the spatial and seasonal variability in quality attributes of wild fish. To do so, wild perch were harvested every three month for a full biological cycle from two different natural systems (Lake Geneva in France and the River Rhine in the Netherlands). Two components of quality were taken into account: organoleptical (colour of skin and fillets (L*C*H° system; 12 variables), seven morphological measurements) and six technological components (viscero, hepato and gonadosomatic index, perivisceral fat index, fillet yield and condition factor K2). Using Principal Component Analysis, we show that within‐season variability is high, particularly for Rhine perch. Perch from the Rhine have a compact body and caudal fin, and possess a longer mouth and head than perch from Lake Genva. Reared fish possess wild‐like characteristics except for technological variables. Fish from extensive and semi‐extensive system show similar attributes, while fish from intensive rearing systems show a higher viscerosomatic and perivisceral fat index.
A comparative study of the performance of family groups of Atlantic salmon reared in tanks and released in a river
Authors: Skilbrei O. T. , Wennevik V.
Published: December 2004

A study was conducted in in the Dale River as a part of a stock enhancement programme. The aim was to compare growth and the incidence of precocious maturation between offspring from precocious and large maturing males, and to study genotype‐environment interactions. River and hatchery performance was compared for 5 × 2 maternal half‐sib family groups, which were stocked as 0+ juveniles or maintained in the hatchery throughout. To identify the offspring, the broodfish were characterized by DNA‐fingerprinting (eight microsatellite markers). Smolt size of 1+ hatchery‐reared smolt and fish caught in a smolt trap, and the size and incidence of precocious maturation among 1+ hatchery‐reared parr and 1+ and 2+ parr caught in the river are compared between the families.
Population genetics, conservation and evolution in salmonids and other widely cultured fishes: some perspectives over six decades
Authors: Utter F.
Published: December 2004

This paper explores my shifting understandings of interactions primarily between salmonid fish culture and fish conservation during the latter half of the 20th century. The idea that conspecific natural and cultured fish were largely interchangeable among phenotypically similar populations began to change with the advent of molecular genetic markers. With the gradual clarification of major geographic lineages beginning in the 1970s came awareness that translocations among anadromous lineages were generally destined for failure; in contrast, gene flow more readily occurred among non‐anadromous lineages and sometimes, species. Concurrently, data were accumulating that adaptations to their respective environments distinguished cultured and wild populations within a lineage. Reduced obstacles to gene flow at this level often resulted in homogenizations among wild and cultured fish in areas where widespread hatchery releases occurred; conversely, adaptive radiations in vacant habitats sometimes occurred over a few decades from single source hatchery releases. Current ideas relating to salmonid interbreeding, population substructure and culture germinated from these observations. Among lineages, resistance to gene flow is much greater between anadromous than purely freshwater populations or species. Ease of within‐lineage gene flow in both groups is problematical with regard to cultured and wild populations because large‐scale supplementation programs erode local adaptations and fine‐scale population substructures. A potential ability to regenerate natural substructure upon relaxation of supplementation is offset by uncertainties of time scales and intrinsic capabilities homogenized populations. However these losses can be minimized by a management that separates harvest and reproduction of wild and cultured subpopulations. Some generality of this strategy to other fishes is supported by losses of local adaptations and outbreeding depression in black basses following population admixtures that parallel those observed in salmonids.
Muscle development in cultured blackspot seabream Pagellus bogaraveo: preliminary histochemical and immunohistochemical data on the fibre types
Authors: Rowlerson A. M. , Silva P. , Rocha E. , Olmedo M. , Valente L. M. P.
Published: December 2004

We studied muscle ontogeny and fibre type characteristics in the blackspot seabream, a new species for commercial aquaculture. Myosin ATPase and SDH histochemistry and immunohistochemistry were tested at different ontogenetic stages, using a panel of antibodies to myosin isoforms and parvalbumin. In general, deep white muscle was parvalbumin‐positive, and superficial ‘red’ muscle was parvalbumin‐negative at all ages examined. At 6 days of age (transition from endogenous to exogenous feeding) three layers of muscle fibres were observed with different antimyosin reactivities: superficial monolayer, presumptive slow red (present only as a small group of fibres adjacent to the lateral line nerve), and presumptive fast‐white (forming the bulk of the muscle). The superficial monolayer and presumptive slow fibres were positive for SDH. At 60 days of age (transition from live to artificial feeding) an additional fibre type was identified: a typical ‘pink’ or intermediate layer. In juveniles, the axial muscle consisted mainly of fast white fibres covered by a slow‐red layer and between them a pink layer. Surprisingly, the red layer could be resolved into two distinct types by myosin immunostaining. Red fibres were also present along the horizontal septum, near the notochord. Both red and white muscle layers showed a mosaic appearance, which was confirmed by ATPase reaction. The work was financed by British Council, CRUP, and FCT (PhD Grant SFRH‐BD‐14068–2003).
Border Effects in Passenger Air Traffic
Authors: Klodt Henning
Published: December 2004

The empirical literature on border effects suggests that national borders substantially reduce the level of economic transactions. This paper adds another piece of evidence to the significance of border effects by applying a new data set which is completely independent of the data sets of previous studies. Our data refer to domestic and international passenger departures from German airports. The econometric results indicate that the German border reduces passenger air traffic by a factor of four to five. This magnitude is in line with the results of previous studies for trade and investment flows.
Consensual or Dominant Relationships with Parliament? A Comparison of Administrations and Ministers in Scotland
Authors: Shephard Mark , Cairney Paul
Published: December 2004

The study of administrations and ministers and their relationships with UK Parliaments has tended to focus on the issues of accountability and responsibility, levels of legislative dissent or broad performance indicators supported by anecdotal examples. This paper addresses the lack of systematic analysis of executive/legislative relations in the policy‐making process by examining the dominance of different administrations and ministers in the Scottish Parliament. Two questions are addressed. First, is there any variance in the legislative dominance of different administrations in the parliamentary arena? Second, do individual ministers make a difference to the degree of policy dominance? Controlling for both initial authorship and quality of amendments to Executive policy, we analyse the nature and extent of Executive dominance during the legislative process of the First Session of the Scottish Parliament. We find some evidence to suggest that Executive dominance varies both by administration and by individual minister.
Reform Styles of Political and Administrative Elites in Majoritarian and Consensus Democracies: Public Management Reforms in New Zealand and the Netherlands
Authors: Yesilkagit Kutsal , De Vries Jouke
Published: December 2004

One of the most important claims advanced by comparative public management reform studies is that these reforms have been more successfully implemented in majoritarian than in consensus democracies. The claim is built on the premise that the institutional structure of a majoritarian democracy enables a parliamentary majority to implement policies unilaterally and in a desired way whereas the institutional structure of a consensus democracy forces parliamentary majorities to negotiate compromises. This claim, we argue, lacks sufficient empirical backing, is biased by studies focusing on the initiation of reforms, and needs serious rethinking in the light of recent empirical studies that downplay the salience of institutions as an explanatory factor for variations in policy reform. With The Netherlands and New Zealand as cases, this article first shows that far‐reaching reforms can be implemented in consensus systems too; secondly, it develops a model that centres on the interplay between institutions and politicians’ reform styles and applies this to these countries.
Coping Behaviors, Parenting, and Perceptions of Children's Internalizing and Externalizing Problems in Rural African American Mothers
Authors: Smith Bynum Mia , H. Brody Gene
Published: December 2004

Abstract: We tested a hypothetical model linking maternal education and maternal coping behavior with parent‐child relationship quality, and in turn, children's self‐regulatory behavior and mental health difficulties. Consistent with predictions, mothers’ use of active coping behaviors predicted more positive parent‐child relationship quality, greater child self‐regulatory behavior, and fewer perceived behavioral and emotional difficulties in children. Implications of these findings for intervention with rural African American mothers are discussed.
Best‐selling Books Advising Parents about Gender: A Feminist Analysis
Authors: Krafchick JenniferL. , Schindler Zimmerman Toni , Haddock ShelleyA. , Banning JamesH.
Published: December 2004

Abstract:To determine how gender in parenting was addressed, a content analysis was conducted on the six best‐selling self‐help books (1997–2002) that provide general parenting advice. A feminist perspective was used to code gendered meaning units in the six books. Findings were that 82% of the implicit gender messages across all books were stereotypical, and thatParenthood by Proxy(Schlessinger, 2000) andChildren Are From Heaven(Gray, 1999) had a gendered agenda in that they contained highly prescriptive content about gender. Practitioners can use these findings in selecting parenting advice books to increase their understanding of the information about gender that parents may glean from these books, and to address these messages with families with whom they work.
Professional Identity and Coping Behaviors in Dual‐Career couples
Authors: Bird GloriaW. , Schnurman‐Crook Abrina
Published: December 2004

Abstract: This qualitative study of 15 dual‐career couples examines the connection between partners’ professional identity and coping behaviors implemented in response to work and family stressors. The analysis provided evidence that dual‐career couples enact professional and family identities that rely on being competent and responsible in both work and family roles. Coping patterns fell into similar patterns across work and family domains, but strategy use was unique by domain and reflective of couples’ belief in the value of interdependent interactions.
Concurrent activation of cell death‐regulating signaling pathways by singlet oxygen in Arabidopsis thaliana
Authors: Danon Antoine , Miersch Otto , Felix Georg , op den Camp Roel G.L. , Apel Klaus
Published: December 2004

Upon a dark/light shift the conditional flu mutant of Arabidopsis starts to generate singlet oxygen (1O2), a non‐radical reactive oxygen species that is restricted to the plastid compartment. Immediately after the shift, plants stop growing and develop necrotic lesions. We have established a protoplast system, which allows detection and characterization of the death response in flu induced by the release of 1O2. Vitamin B6 that quenches 1O2 in fungi was able to protect flu protoplasts from cell death. Blocking ethylene production was sufficient to partially inhibit the death reaction. Similarly, flu mutant seedlings expressing transgenic NahG were partially protected from the death provoked by the release of 1O2, indicating a requirement for salicylic acid (SA) in this process, whereas in cells depleted of both, ethylene and SA, the extent of cell death was reduced to the wild‐type level. The flu mutant was also crossed with the jasmonic acid (JA)‐depleted mutant opr3, and with the JA, OPDA and dinor OPDA (dnOPDA)‐depleted dde2‐2 mutant. Analysis of the resulting double mutants revealed that in contrast to the JA‐induced suppression of H2O2/superoxide‐dependent cell death reported earlier, JA promotes singlet oxygen‐mediated cell death in flu, whereas other oxylipins such as OPDA and dnOPDA antagonize this death‐inducing activity of JA.
New human and mouse microRNA genes found by homology search
Authors: Weber Michel J.
Published: December 2004

Conservation of microRNAs (miRNAs) among species suggests that they bear conserved biological functions. However, sequencing of new miRNAs has not always been accompanied by a search for orthologues in other species. I report herein the results of a systematic search for interspecies orthologues of miRNA precursors, leading to the identification of 35 human and 45 mouse new putative miRNA genes. MicroRNA tracks were written to visualize miRNAs in human and mouse genomes on the UCSC Genome Browser. Based on their localization, miRNA precursors can be excised either from introns or exons of mRNAs. When intronic miRNAs are antisense to the apparent host gene, they appear to originate from ill‐characterized antisense transcription units. Exonic miRNAs are, in general, nonprotein‐coding, poorly conserved genes in sense orientation. In three cases, the excision of an miRNA from a protein‐coding mRNA might lead to the degradation of the rest of the transcript. Moreover, three new examples of miRNAs fully complementary to an mRNA are reported. Among these, miR135a might control the stability and/or translation of an alternative form of the glycerate kinase mRNA by RNA interference. I also discuss the presence of human miRNAs in introns of paralogous genes and in miRNA clusters.
The role of helix 8 and of the cytosolic C‐termini in the internalization and signal transduction of B1 and B2 bradykinin receptors
Authors: Faussner Alexander , Bauer Alexandra , Kalatskaya Irina , Schüssler Steffen , Seidl Cornelia , Proud David , Jochum Marianne
Published: December 2004

Determinants for desensitization and sequestration of G protein‐coupled receptors often contain serine or threonine residues located in their C‐termini. The sequence context, however, in which these residues have to appear, and the receptor specificity of these motifs are largely unknown. Mutagenesis studies with the B2 bradykinin receptor (B2wt), stably expressed in HEK 293 cells, identified a sequence distal to N338 (NSMGTLRTSI, including I347 but not the basally phosphorylated S348) and in particular the TSI sequence therein, as a major determinant for rapid agonist‐inducible internalization and the prevention of receptor hypersensitivity. Chimeras of the noninternalizing B1 bradykinin receptor (B1wt) containing these B2wt sequences sequestered poorly, however, suggesting that additional motifs more proximal to N338 are required. In fact, further substitution of the B1wt C‐terminus with corresponding B2wt regions either at C330(7.71) following putative helix 8 (B1CB2) or at the preceding Y312(7.53) in the NPXXY sequence (B1YB2) resulted in chimeras displaying rapid internalization. Intriguingly, however, exchange performed at K322(7.63) within putative helix 8 generated a slowly internalizing chimera (B1KB2). Detailed mutagenesis analysis generating additional chimeras identified the change of V323 in B1wt to serine (as in B2wt) as being responsible for this effect. The slowly internalizing chimera as well as a B1wt point‐mutant V323S displayed significantly reduced inositol phosphate accumulation as compared to B1wt or the other chimeras. The slow internalization of B1KB2 was also accompanied by a lack of agonist‐induced phosphorylation, that in contrast was observed for B1YB2 and B1CB2, suggesting that putative helix 8 is either directly or indirectly (e.g. via G protein activation) involved in the interaction between the receptor and receptor kinases.
Pleiotropy of leptin receptor signalling is defined by distinct roles of the intracellular tyrosines
Authors: Hekerman Paul , Zeidler Julia , Bamberg‐Lemper Simone , Knobelspies Holger , Lavens Delphine , Tavernier Jan , Joost Hans‐Georg , Becker Walter
Published: December 2004

The leptin receptor (LEPR) is a class I cytokine receptor signalling via both the janus kinase/signal transducer and activator of transcription (JAK/STAT) and the MAP kinase pathways. In addition, leptin has been shown previously to activate AMP‐activated kinase (AMPK) in skeletal muscle. To enable a detailed analysis of leptin signalling in pancreatic beta cells, LEPR point mutants with single or combined exchanges of the three intracellular tyrosines were expressed in HIT‐T15 insulinoma cells. Western blots with activation state‐specific antibodies recognizing specific signalling molecules revealed that the wild‐type receptor activated STAT1, STAT3, STAT5 and ERK1/2 but failed to alter the phosphorylation of AMPK. Each of the three intracellular tyrosine residues in LEPR exhibited different signalling capacities: Tyr985 was necessary and sufficient for leptin‐induced activation of ERK1/2; Tyr1077 induced tyrosyl phosphorylation of STAT5; and Tyr1138 was capable of activating STAT1, STAT3 and STAT5. Consistent results were obtained in reporter gene assays with STAT3 or STAT5‐responsive promoter constructs, respectively. Furthermore, the sequence motifs surrounding the three tyrosine residues are conserved in LEPR from mammals, birds and in a LEPR‐like cytokine receptor from pufferfish. Mutational analysis of the box3 motif around Tyr1138 identified Met1139 and Gln1141 as important determinants that define specificity towards the different STAT factors. These data indicate that all three conserved tyrosines are involved in LEPR function and define the pleiotropy of signal transduction via STAT1/3, STAT5 or ERK kinases. Activation and inhibition of AMPK appears to require additional components of the signalling pathways that are not present in beta cells.
Androgen receptor function is modulated by the tissue‐specific AR45 variant
Authors: Ahrens‐Fath Isabelle , Politz Oliver , Geserick Christoph , Haendler Bernard
Published: December 2004

A naturally occurring variant of the human androgen receptor (AR) named AR45 has been identified. It lacks the entire region encoded by exon 1 of the AR gene and is composed of the AR DNA‐binding domain, hinge region and ligand‐binding domain, preceded by a novel seven amino‐acid long N‐terminal extension. A survey of human tissues revealed that AR45 was expressed mainly in heart and skeletal muscle. In cotransfection experiments, AR45 inhibited AR function, an effect necessitating intact DNA‐ and ligand‐binding properties. Overexpression of AR45 reduced the proliferation rate of the androgen‐dependent LNCaP cells, in line with the repressive effects of AR45 on AR growth‐promoting function. AR45 interacted with the AR N‐terminal domain in two‐hybrid assays, suggesting that AR inhibition was due to the formation of AR–AR45 heterodimers. Under conditions where the transcriptional coactivator TIF2 or the oncogene β‐catenin were overexpressed, AR45 stimulated androgen‐dependent promoters in presence of dihydrotestosterone. AR45 activity was triggered additionally by the adrenal androgen androstenedione in presence of exogenous TIF2. Altogether, the data suggest an important role of AR45 in modulating AR function and add a novel level of complexity to the mode of action of androgens.
Conformational and functional analysis of the lipid binding protein Ag‐NPA‐1 from the parasitic nematode Ascaridia galli
Authors: Jordanova Rositsa , Radoslavov Georgi , Fischer Peter , Liebau Eva , Walter Rolf D. , Bankov Ilia , Boteva Raina
Published: December 2004

Ag‐NPA‐1 (AgFABP), a 15 kDa lipid binding protein (LBP) from Ascaridia galli, is a member of the nematode polyprotein allergen/antigen (NPA) family. Spectroscopic analysis shows that Ag‐NPA‐1 is a highly ordered, α‐helical protein and that ligand binding slightly increases the ordered secondary structure content. The conserved, single Trp residue (Trp17) and three Tyr residues determine the fluorescence properties of Ag‐NPA‐1. Analysis of the efficiency of the energy transfer between these chromophores shows a high degree of Tyr‐Trp dipole‐dipole coupling. Binding of fatty acids and retinol was accompanied by enhancement of the Trp emission, which allowed calculation of the affinity constants of the binary complexes. The distance between the single Trp of Ag‐NPA‐1 and the fluorescent fatty acid analogue 11‐[(5‐dimethylaminonaphthalene‐1‐ sulfonyl)amino]undecanoic acid (DAUDA) from the protein binding site is 1.41 nm as estimated by fluorescence resonance energy transfer. A chemical modification of the Cys residues of Ag‐NPA‐1 (Cys66 and Cys122) with the thiol reactive probes 5‐({[(2‐iodoacetyl)amino]ethyl}amino) naphthalene‐1‐sulfonic acid (IAEDANS) and N,N′‐dimethyl‐N‐(iodoacetyl)‐N′‐(7‐nitrobenz‐2‐oxa‐1,3‐diazol‐4‐yl)ethylenediamine (IANBD), followed by MALDI‐TOF analysis showed that only Cys66 was labeled. The observed similar affinities for fatty acids of the modified and native Ag‐NPA‐1 suggest that Cys66 is not a part of the protein binding pocket but is located close to it. Ag‐NPA‐1 is one of the most abundant proteins in A. galli and it is distributed extracellularly mainly as shown by immunohistology and immunogold electron microscopy. This suggests that Ag‐NPA‐1 plays an important role in the transport of fatty acids and retinoids.
The Rab5 effector Rabaptin‐5 and its isoform Rabaptin‐5δ differ in their ability to interact with the small GTPase Rab4
Authors: Korobko Elena , Kiselev Sergey , Olsnes Sjur , Stenmark Harald , Korobko Igor
Published: December 2004

Rabaptin‐5 is an effector for the small GTPase Rab5, a regulator of the early steps in endocytosis. In addition, Rabaptin‐5 interacts with the small GTPase Rab4 that has been implicated in recycling from early endosomes to the cell surface. Recently we have identified a ubiquitous transcript encoding the Rabaptin‐5 isoform, Rabaptin‐5δ. To evaluate the interaction properties of Rabaptin‐5δ with the small GTPases Rab4 and Rab5, we have applied protein interaction assays using the yeast two‐hybrid system and a glutathione S‐transferase pull‐down assay. We found that unlike Rabaptin‐5, that interacts with both GTPases in GTP‐bound conformations, Rabaptin‐5δ interacts only with GTP‐bound Rab5, and does not interact with Rab4, presumably due to a disrupted Rab4 binding site. Immunofluorescence microscopy analysis carried out to address the localization of Rabaptin‐5δ relative to GTP‐bound Rab4 and Rab5 in BHK‐21 cells supported these data. Our data suggests that while Rabaptin‐5 was proposed to act as a molecular linker between Rab5 and Rab4, to coordinate endocytic and recycling traffic, Rabaptin‐5δ is involved only in the Rab5‐driven events.
Kinetics of intra‐ and intermolecular zymogen activation with formation of an enzyme–zymogen complex
Authors: Fuentes Matilde Esther , Varón Ramón , García‐Moreno Manuela , Valero Edelmira
Published: December 2004

A mathematical description was made of an autocatalytic zymogen activation mechanism involving both intra‐ and intermolecular routes. The reversible formation of an active intermediary enzyme–zymogen complex was included in the intermolecular activation route, thus allowing a Michaelis–Menten constant to be defined for the activation of the zymogen towards the active enzyme. Time–concentration equations describing the evolution of the species involved in the system were obtained. In addition, we have derived the corresponding kinetic equations for particular cases of the general model studied. Experimental design and kinetic data analysis procedures to evaluate the kinetic parameters, based on the derived kinetic equations, are suggested. The validity of the results obtained were checked by using simulated progress curves of the species involved. The model is generally good enough to be applied to the experimental kinetic study of the activation of different zymogens of physiological interest. The system is illustrated by following the transformation kinetics of pepsinogen into pepsin.
Towards discovery‐driven translational research in breast cancer
Authors: Celis Julio E. , Moreira José M. A. , Gromova Irina , Cabezon Teresa , Ralfkiaer Ulrik , Guldberg Per , Straten Per thor , Mouridsen Henning , Friis Esbern , Holm Dorte , Rank Fritz , Gromov Pavel
Published: December 2004

Discovery‐driven translational research in breast cancer is moving steadily from the study of cell lines to the analysis of clinically relevant samples that, together with the ever increasing number of novel and powerful technologies available within genomics, proteomics and functional genomics, promise to have a major impact on the way breast cancer will be diagnosed, treated and monitored in the future. Here we present a brief report on long‐term ongoing strategies at the Danish Centre for Translational Breast Cancer Research to search for markers for early detection and targets for therapeutic intervention, to identify signalling pathways affected in individual tumours, as well as to integrate multiplatform ‘omic’ data sets collected from tissue samples obtained from individual patients. The ultimate goal of this initiative is to coalesce knowledge‐based complementary procedures into a systems biology approach to fight breast cancer.
Inhalation of tobramycin in a patient with aspiration pneumonia as a result of medullary stroke
Authors: OHWADA Akihiko , KAJIYAMA Yuichiro , HIRASHIMA Naoko , MIYAJI Atsuko , SUZUKI Masaru
Published: December 2004

Abstract:  A 78‐year‐old man suffered from refractory aspiration pneumonia as a result of a minor medullary stroke. The only neurological symptom observed in this patient was difficulty in swallowing. He was managed with i.v. hyperalimentation with termination of oral intake, including water. However, he still experienced several episodes of aspiration pneumonia. As he was considered to have a bacterial infection because of silent aspiration of colonized oropharyngeal material, inhalation of tobramycin was introduced and successful control of airway infection was attained.
Comparison of patterns of care in lung cancer in three area health services in New South Wales, Australia
Authors: Vinod S. K. , Hui A. C. , Esmaili N. , Hensley M. J. , Barton M. B.
Published: December 2004

Background: Lung cancer is the leading cause of cancer deaths in New South Wales (NSW). The incidence of and mortality from lung cancer differ throughout different area health services in NSW.

Aim: To compare patterns of care in lung cancer among three area health services in NSW.

Methods: South‐western Sydney Area Health Service (SWSAHS), Northern Sydney Area Health Service (NSAHS) and Hunter Area Health Service (HAHS) ­residents diagnosed with lung cancer in 1996 were identified from the NSW Central Cancer Registry and their medical records were reviewed. The main outcome measures were specialist care, investigations, treatment and survival.

Results: The study population comprised 256 SWSAHS, 270 NSAHS and 212 HAHS residents. NSAHS residents were older, with a median age of 73 years compared with 68 years in SWSAHS and 70 years in HAHS (P = 0.001). The performance status and stage distributions of the populations were similar. Twenty per cent of HAHS residents did not have a pathological diagnosis compared with 10% in SWSAHS and 9% in NSAHS (P = 0.005). Forty‐five per cent of HAHS residents received no treatment compared with 25 and 22% in SWSAHS and NSAHS, respectively (P < 0.001). Despite these differences, there was no significant difference in overall survival.

Conclusions: Lung cancer patterns of care were significantly different among the areas. The variability of practice identified in this study needs to be addressed to ensure optimum care for all patients with lung cancer. Although there was no significant difference in survival, under‐utilization of efficacious treatment is likely to have affected patients’ quality of life. (Intern Med J 2004; 34: 677−683)
Concordance between clopidogrel use and prescribing guidelines
Authors: Kubler P. A. , Pillans P. I. , Marrinan M. C. , Frogley M.
Published: December 2004

Background: Clopidogrel is an antiplatelet drug increasingly used in the secondary prevention of atherosclerotic vascular events. Compared with aspirin, clopidogrel has marginal additional efficacy and similar safety, but carries a substantial price premium. It remains unclear whether its use is cost‐effective.

Aims: (i) To determine concordance between clopidogrel use and the Pharmaceutical Benefits Scheme and local hospital prescribing guidelines and (ii) to determine the intended duration of clopidogrel therapy and ascertain whether this is supported by other published reports.

Methods: Cross‐sectional evaluation by patient interview and chart review of appropriateness of clopidogrel prescribing was carried out for 100 consecutive patients attending a 700 bed metropolitan, primary care and tertiary referral hospital.

Results: Clopidogrel was predominately used for secondary prevention of ischaemic heart disease (60%) and following percutaneous coronary intervention (34%). A significant proportion of patients (29%) received clopidogrel outside the prescribing guidelines. Many patients were intended to receive indefinite therapy for secondary prevention of ischaemic vascular events. Concomitant aspirin therapy was not prescribed in 23% of patients, for 78% of whom it was inappropriate.

Conclusions: There is a lack of concordance between clopidogrel use and prescribing guidelines. In the majority of patients (71%), clopidogrel is used for valid indications but there is considerable leakage of use beyond prescribing guidelines. Concomitant aspirin therapy is often not prescribed in the absence of clinically relevant contraindications. Moreover, treatment is continued in many patients beyond what is supported by current published data. (Intern Med J 2004; 34: 663−667)
Mapping environments at risk under different global climate change scenarios
Authors: Saxon Earl , Baker Barry , Hargrove William , Hoffman Forrest , Zganjar Chris
Published: December 2004

All global circulation models based on Intergovernmental Panel on Climate Change (IPCC) scenarios project profound changes, but there is no consensus on how to map their environmental consequences. Our multivariate representation of environmental space combines stable topographic and edaphic attributes with dynamic climatic attributes. We divide that environmental space into 500 unique domains and map their current locations and their projected locations in 2100 under contrasting emissions scenarios. The environmental domains found across half the study area today disappear under the higher emissions scenario, but persist somewhere in it under the lower emissions scenario. Locations affected least and those affected most under each scenario are mapped. This provides an explicit framework for designing conservation networks to include both areas at least risk (potential refugia) and areas at greatest risk, where novel communities may form and where sentinel ecosystems can be monitored for signs of stress.
A Continuous Time Markov Model for the Length of Stay of Elderly People in Institutional Long-Term Care
Authors: Xie H. , Chaussalet T. J. , Millard P. H.
Published: December 2004

The paper develops a Markov model in continuous time for the length of stay of elderly people moving within and between residential home care and nursing home care. A procedure to determine the structure of the model and to estimate parameters by maximum likelihood is presented. The modelling approach was applied to 4 years’ placement data from the social services department of a London borough. The results in this London borough suggest that, for residential home care, a single-exponential distribution with mean 923 days is adequate to provide a good description of the pattern of the length of stay, whereas, for nursing home care, a mixed exponential distribution with means 59 days (short stay) and 784 days (long stay) is required, and that 64% of admissions to nursing home care will become long-stay residents. The implications of these findings and the advantages of the proposed modelling approach in the general context of long-term care are discussed.
An Agency Theory Perspective on Student Performance Evaluation
Authors: Smith Michael E. , Zsidisin George A. , Adams Laural L.
Published: December 2004

The emphasis in recent research on the responsibility of college and university business instructors to prepare students for future employment underscores a need to refine the evaluation of student performance. In this article, an agency theory framework is used to understand the trade‐offs that may be involved in the selection of various approaches to student evaluation. Understanding these trade‐offs may be particularly important as faculty members seek to balance competing obligations, such as research and service requirements, while ensuring instructional effectiveness. This article presents propositions for examining how various institutional, instructor, and student characteristics influence the selection and use of student performance evaluation techniques (i.e., exams, papers, and group assignments). In conclusion, we suggest that agency theory may serve as a foundation for understanding current evaluation practices and guiding instructors in their selection of appropriate evaluation mechanisms.
Using Quality Management Tools to Enhance Feedback from Student Evaluations
Authors: Jensen John B. , Artz Nancy
Published: December 2004

Statistical tools found in the service quality assessment literature—the T2 statistic combined with factor analysis—can enhance the feedback instructors receive from student ratings. T2 examines variability across multiple sets of ratings to isolate individual respondents with aberrant response patterns (i.e., outliers). Analyzing student responses that are outside the “normal” range of responses can identify aspects of the course that cause pockets of students to be dissatisfied. This fresh insight into sources of student dissatisfaction is particularly valuable for instructors willing to make tactical classroom changes that accommodate individual students rather than the traditional approach of using student ratings to develop systemwide changes in course delivery. A case study is presented to demonstrate how the recommended procedure minimizes data overload, allows for valid schoolwide and longitudinal comparisons of correlated survey responses, and helps instructors identify priority areas for instructional improvement.
Orofacial and gastrointestinal hyperplasia and neoplasia in smad4+/− and elf+/−/smad4+/− mutant mice
Authors: Redman Robert S. , Katuri Varalakshmi , Tang Yi , Dillner Allan , Mishra Bibhuti , Mishra Lopa
Published: December 2004

Background:  Smad4 is vital to the roles of Smads 2 and 3 in transforming growth factor‐beta (TGF)‐β signal transduction, and inactivated Smad4 is common to human gastrointestinal cancers. The embryonic liver fodrin (ELF) is a β‐spectrin that facilitates the nuclear translocation of activated Smad4.

Methods:  Smad4
+/− mice, known to develop gastrointestinal cancer, were crossbred with elf+/− mice. The smad4+/− and smad4+/−/elf+/− offspring were autopsied as abnormalities developed.

Results:  In addition to polyps and adenocarcinomas of the stomach and duodenum, the smad4+/− mice developed squamous cell carcinomas of the skin, oral mucosa and forestomach, benign neoplasms of connective tissue and lacrimal gland, and a lymphoma. The smad4+/−/elf+/− mice developed extensive hyperplasia and neoplasia of the gastric mucosa.

Conclusion:  These findings indicate that investigating interactions among smad4, elf, and other genes involved in TGF‐β signaling should be useful in further delineating the processes of neoplasia in a wide variety of tissues.
Immunohistochemical analysis of apoptosis‐related factors in lining epithelium of radicular cysts
Authors: Suzuki T. , Kumamoto H. , Kunimori K. , Ooya K.
Published: December 2004

Background:  Some studies suggest that apoptosis‐related factors are involved in the inflammatory processes of marginal periodontal lesions. However, the role of apoptosis in periapical inflammatory lesions remains unclear. We investigated the possible role of apoptotic cell death in periapical inflammatory lesions by means of immunohistochemical analysis of apoptosis‐related factors and use of a cell proliferation marker.

Methods:  Paraffin‐embedded sections of 19 radicular cysts (RCs), and five residual radicular cysts (RRCs) and control specimens of normal gingivae excised from seven cadavers were prepared and examined immunohistochemically with the use of monoclonal antibodies or polyclonal antisera against single‐stranded DNA (ssDNA), p53, Bax, Bcl‐2, caspase‐3, Fas, Fas ligand (Fas‐L), and Ki‐67 antigen.

Results:  Epithelium of gingiva, RCs, and RRCs showed expression of ssDNA in suprabasal and superficial epithelial cells and Ki‐67 reactivity in basal and parabasal cells. Expression of Ki‐67 and ssDNA in RCs and RRCs was slightly higher than that in gingiva. Both Ki‐67 and ssDNA reactivity in RCs with intense inflammatory reactions or with thick lining epithelium were significantly stronger than those in RCs with less inflammatory reactions or with thin lining epithelium. Reactivity for p53 was noted sporadically in epithelium of gingiva, RCs, and RRCs, and p53 expression in RCs was significantly greater than that in gingiva. Ki‐67 and ssDNA reactivity in RCs increased parallel to the degree of p53 expression. Bax and Bcl‐2 were detected in some basal epithelial cells in RCs and RRCs as well as in gingiva. The ssDNA reactivity significantly increased parallel to Bax expression and slightly decreased parallel to Bcl‐2 expression in lining epithelium of RCs. Caspase‐3 was detected in superficial epithelial cells of both gingiva and lining epithelium of RCs and RRCs, and the distribution of these cells was compatible with the expression of ssDNA. Expression of Ki‐67 and ssDNA in caspase‐3‐positive fields was significantly higher than that in caspase‐3‐negative fields in RCs. There was very limited expression of Fas and Fas‐L in lining epithelium of RCs and RRCs as well as in gingiva.

Conclusions:  These data suggest that apoptosis‐related factors are involved in the pathophysiologic activity of periapical inflammatory lesions. Such factors may be affected by the structure of lining epithelium and the degree of inflammatory change.