bluuebunny/crossref_metadata_2025 huggingface dataset:
Identification of new flaviviruses in the Kokobera virus complex
Authors: Nisbet Debra J. , Lee Katie J. , van den Hurk Andrew F. , Johansen Cheryl A. , Kuno Goro , Chang Gwong-Jen J. , Mackenzie John S. , Ritchie Scott A. , Hall Roy A.
Published: December 2004

Novel flavivirus isolates from mosquitoes collected in northern Australia were analysed by partial genomic sequencing, monoclonal antibody-binding assays and polyclonal cross-neutralization tests. Two isolates were found to be antigenically distinct from, but related to, viruses of the Kokobera virus complex, which currently contains Kokobera (KOKV) and Stratford (STRV) viruses. Nucleotide sequence comparison of two separate regions of the genome revealed that an isolate from Saibai Island in the Torres Strait in 2000 (TS5273) was related closely to KOKV and STRV, with 74–80 and 75–76 % nucleotide similarity, respectively. An isolate from mainland Cape York in 1998 (CY1014) was found to be more divergent from KOKV and STRV, with <70 % nucleotide sequence similarity to either virus. It is proposed that isolate TS5273 represents a new subtype of KOKV and that CY1014 be classified as a novel species within the Kokobera virus complex of flaviviruses, named New Mapoon virus.
Heterogeneity of hepatitis C virus genotype 4 strains circulating in south-western France
Authors: Nicot Florence , Legrand-Abravanel Florence , Sandres-Saune Karine , Boulestin Anne , Dubois Martine , Alric Laurent , Vinel Jean-Pierre , Pasquier Christophe , Izopet Jacques
Published: December 2004

Hepatitis C virus (HCV) is a major cause of liver disease. Knowledge of HCV variability is crucial for clinical and epidemiological analysis. HCV genotype 4 (HCV-4) has become increasingly prevalent in European countries, including France, in recent years. The present study investigates the heterogeneity of HCV-4 in south-western France by phylogenetic analysis of NS5B sequences from 166 patients. The E2 region of 90 strains was also analysed. Genotype 4 accounts for 7·4 % of HCV infections in this area. Analysis of the NS5B region revealed 12 subtypes and the NS5B and E2 phylogeny data were congruent, except for one strain. The epidemiological data indicated two main groups of patients. One included intravenous drug users (IVDUs) of French origin, who were infected by homogeneous strains of subtypes 4a or 4d. The second group comprised non-IVDU patients who were infected with heterogeneous strains. This group was subdivided into patients of French origin, who were infected with eight subtypes, and patients from non-European countries (Central Africa or the Middle East), who were mainly infected with 4f, 4k, 4r and other subtypes; they showed the greatest genetic heterogeneity. This study of a large cohort of patients shows the great diversity of HCV-4 strains, and that these subtypes have spread differently.
Murine gammaherpesvirus 68 bcl-2 homologue contributes to latency establishment in vivo
Authors: de Lima Brigitte D. , May Janet S. , Marques Sofia , Simas J. Pedro , Stevenson Philip G.
Published: December 2004

The gammaherpesviruses are characteristically latent in lymphocytes and exploit lymphocyte proliferation to establish a large, persistent pool of latent genomes. Murine gammaherpesvirus 68 (MHV-68) allows the in vivo analysis of viral genes that contribute to this and other aspects of host colonization. In this study, the MHV-68 bcl-2 homologue, M11, was disrupted either in its BH1 homology domain or upstream of its membrane-localizing C-terminal domain. Each M11 mutant showed normal lytic replication in vitro and in vivo, but had a reduction in peak splenic latency. Lower infectious-centre titres correlated with lower in vivo B-cell activation, lower viral genome loads and reduced viral tRNA expression. This was therefore a true latency deficit, rather than a deficit in ex vivo reactivation. Stable, long-term levels of splenic latency were normal. M11 function therefore contributed specifically to viral latency amplification in infected lymphoid tissue.
Hepatitis C virus F protein sequence reveals a lack of functional constraints and a variable pattern of amino acid substitution
Authors: Cristina Juan , Lopez Fernando , Moratorio Gonzalo , López Lilia , Vasquez Silvia , García-Aguirre Laura , Chunga Ausberto
Published: December 2004

Hepatitis C virus (HCV) is an important human pathogen that affects 170 million people worldwide. The HCV genome is an RNA molecule that is approximately 9·6 kb in length and encodes a polyprotein that is cleaved proteolytically to generate at least 10 mature viral proteins. Recently, a new HCV protein named F has been described, which is synthesized as a result of a ribosomal frameshift. Little is known about the biological properties of this protein, but the possibility that the F protein may participate in HCV morphology or replication has been raised. In this work, the presence of functional constraints in the F protein was investigated. It was found that the rate of amino acid substitutions along the F protein was significantly higher than the rate of synonymous substitutions, and comparisons involving genes that represented independent phylogenetic lineages yielded very different divergence/conservation patterns. The distribution of stop codons in the F protein across all HCV genotypes was also investigated; genotypes 2 and 3 were found to have more stop codons than genotype 1. The results of this work suggest strongly that the pattern of divergence in the F protein is not affected by functional constraints.
Determinants of women's choice of their obstetrician and gynecologist provider in the UAE
Authors: Rizk Diaa E. E. , El‐Zubeir Margaret A. , Al‐Dhaheri Alia M. , Al‐Mansouri Fatima R. , Al‐Jenaibi Houda S.
Published: December 2004

Background.  The objective of this study is to evaluate women's priorities and preferences in selecting their obstetrician and gynecologist in a non‐Western society.

Methods.  Consecutive sample of 508 patients attending the obstetric and gynecologic services of AL‐Ain Hospital; AL‐Ain; United Arab Emirates during 4 months was recruited. Participants were interviewed by using a structured 26‐item questionnaire administered by means of face‐to‐face interview within 24 h of admission in the ward or before consultation in the outpatient clinic. Respondents were asked about their preferences, priorities, determinants of choice of their obstetrician and gynecologist, their view of his/her role as a medical provider, and their perception of importance of each of provider's characteristics and roles on a five‐point Likert scale.

Results.  Four hundred thirty‐nine (86.4%) participants preferred female physicians. Sixty‐one (12%) had no preference and eight (1.6%) preferred male physicians. Reasons for female selection were privacy during intimate examination (89·1%) or counseling (68.8%), religious beliefs (74.3%), and cultural traditions (45.3%). Female preference was significantly associated with higher parity (P = 0.002), religion (P = 0.005), nationality (P = 0.01), occupation (P = 0.02), education (P = 0.04), and poor recognition of physician's role as professional/expert (P < 0.00001). Male preference was significantly associated with experience at previous encounter (P = 0.03), obstetric consultation (P = 0.04), and perceiving physician's role as skilled communicator (P = 0.01) or health educator (P = 0.04). Other physician characteristics affecting choice were professionalism, bedside manners, empathy, communication, competence, availability, and religion.

Conclusions.  Physician's attitude, professional profile, sex, and religious faith determine women's choice of obstetricians and gynecologists. Most women prefer female providers because of embarrassment during pelvic examination and reproductive counseling, religious beliefs, and sociocultural values.
Variable number of tandem repeats in the growth hormone gene of Sparus aurata: association with growth and effect on gene transcription
Authors: Almuly R. , Poleg‐Danin Y. , Kashi Y. , Gorshkov S. , Gorshkova G. , Dyman A. , Cavari B. , Rapoport B. , Soller M. , Funkenstein B.
Published: December 2004

The GH gene of Sparus aurata(saGH) contains variable number of tandem repeats (VNTR). The hyper‐variable minisatellites in the first and third introns segregate in a Mendelian manner and exhibit numerous alleles. Analysis by PCR and sequencing of the two introns in several wild Sparidae species revealed comparable minisatellites with some variations. ‘Zoo blot’ with the first intron unit as a probe showed this sequence to be characteristic of several families from the Perciformes order. Unexpectedly, a similar minisatellite was found in the first intron of the GH gene in flounder, which belongs to a different order. Transfection of constructs containing a reporter gene and first intron of different length to four cell lines resulted in an inhibitory effect of the longer intron relative to the short intron. A (CA)n microsatellite (saGHpCA) is found in the GH promoter. A similar repeat at the same location is present in GH promoters of several other fish species. High variability (11 alleles) of the saGHpCA was found in a hatchery population. Full‐sib family genotyping showed a Mendelian inheritance of these alleles. A significant association was found between allele distribution and body mass in large and average size fishes from a hatchery population. The intron minisatellites may serve as markers for hybrid population and parental assignment. Its presence in families and orders of the higher teleosts may help solving classification uncertainties. Their conservation and inhibitory effect suggest a biological role. The saGHpCA is correlated with growth and may be a good candidate for predicting growth performance.
Changes of nase Chondrostoma nasus L. occurrence in the Wislok River, southern Poland, after stocking with pond reared juveniles
Authors: Ciesla M. , Sliwinski J. , Konieczny P. , Litynski R.
Published: December 2004

Nase Chondrostoma nasus L., once one of the most common fish species in the upper reaches of central European rivers, is now considered endangered throughout its natural range. Since 2000 the Department of Ichthyobiology and Fisheries, Warsaw Agricultural University, has co‐operated with the Polish Anglers Association in Krosno to study the effects of river restocking with pond‐farmed nase juveniles. The experiment is carried out on Wislok River, where nase was very twenty years ago but has now completely disappeared. Selected parts of the river are stocked with tagged, one‐summer and 1 year‐old juveniles. Changes in nase occurrence, and their growth and foraging behaviour have been analysed.
What are the reasons for the Prussian carp expansion in the upper Elbe River, Czech Republic?
Authors: Slavík O. , Bartoš L.
Published: December 2004

Fifty five Prussian carp Carassius auratus gibelio were radio‐tracked weekly along the course of the upper Elbe River, for 15 months. In addition, diurnal movements were observed in three different habitat types (main channel, navigation canal and floodplain areas). Fish species composition from 10 fish ladders was regularly analysed in 1996, 2000 and 2001, and patterns of juvenile (0+ year) fish distribution and abundance were collected in 2000 and 2001, to support telemetric data on longitudinal migrations, and habitat preference within the studied stretch. Downstream migrations dominated in all seasons, being most intensive in spring and reaching a maximum distance of 85 km. Migration stopped when specimens found a floodplain area, where they occupied larger home ranges compared to the main channel. Migrations upstream of up to 2 km were rare, and specimens did not enter fish ladders. Spontaneous reproduction only occurred on the floodplain and its success was very low. The results suggested that neither upstream migrations, nor spontaneous reproduction represented the key mechanism for the Prussian carp invasion into the Elbe catchment area of the Czech Republic. It can be assumed that escapes from aquaculture activities are primarily responsible for its expansion.
Maladaptation and phenotypic mismatch in cultured Atlantic salmon used for stocking
Authors: García de Leániz C. , Consuegra S. , Serdio A.
Published: December 2004

Compared to wild populations, cultured Atlantic salmon often sustain higher mortality rates and lower adult return rates when stocked as juveniles into natural streams. The ultimate causes for such differences in fitness, however, are not clear. Here we tested if relaxed levels of natural selection and improved survival in the hatchery environment could account for the observed degree of maladaptation among stocked fish. To do this, we assessed the degree of phenotypic mismatch between wild and cultured fish in three populations over five consecutive years. Significant differences were found in several phenotypic traits that are likely to have fitness implications. Thus, if the objective is to mimic wild individuals for restoration purposes, current hatchery practices aimed at maximising juvenile survival and enhancing growth may need to be revised.
Efficiency of allele frequency‐based bayesian programmes for detecting hybridization between farmed and wild salmon
Authors: Vähä J.‐P. K. , Primmer C. R.
Published: December 2004

Large escapes of cultured salmon from net‐pens have become inevitable disasters linked to the growth of aquaculture in coastal areas. Hybridization between farmed and wild salmon has been witnessed; but the extent of eventual genetic introgression is controversial as selection against hybrids can maintain distinct gene pools. Individual assignment tests based on genetic data have been widely used in fisheries, due to the importance of accurate population assignment for a variety of purposes including distinction between individuals of native and stocked origin. However the ability of these Bayesian programs to detect hybrids and subsequent generations between closely related populations has been little investigated. Here we present results regarding the efficiency of two new computer programs, structure and New Hybrids in detecting hybridization between farmed and wild salmon from the river Teno (Northern Europe) based on genetic data obtained from 17 microsatellite loci.
Disruption of the melatonin rhythm in wild and farmed fish: a consequence of prolonged thyroxine administration and calcium depletion
Authors: Kulczykowska E. , Sokolowska E. , Gozdowska M. , Kalamarz H.
Published: December 2004

Recent studies suggest that the pineal gland and its major product melatonin (N‐acetyl‐5‐methoxytryptamine: Mel) play a role in integration of various neural and endocrine functions. Observations indicate that Mel as a signal of photoperiod regulates a number of biological phenomena, including reproduction, day/night activity and many other physiological events associated with daily or seasonal rhythms in vertebrates, including fish. Mel has also been found to be a highly effective preventive antioxidant and free radical scavenger, protecting organism from oxidative damage. In all species examined, plasma melatonin concentration shows a diurnal rhythm, with the highest levels during the night. Our studies are focused on the cues affecting Mel synthesis capacity in wild and farmed fish. The Mel rhythm in fish can be disrupted by fluctuations in surrounding calcium concentration or by prolonged thyroxine (T4) exposure. Physiological depletion of plasma ionized calcium may limit the capacity of Mel night production in two fish species, farmed rainbow trout and wild flounder. Prolonged (2 week) exposure to high level of exogenous thyroxine (T4) results in reduced night‐time plasma Mel levels, and may thus inhibit the melatonin‐related time‐keeping system in juvenile salmon. Disruption of the daily Mel rhythm implies a reduced ability of organisms to respond properly to environmental signals, and may be a useful indicator of disturbance in physiological functions.
Coase Theorem and the Taiwan Strait Conflict
Authors: Hsiung Bingyuan
Published: December 2004

The idea of a single‐owner who owns the opposing interests of a dispute has been suggested in the literature to make the Coase Theorem operational in resolving dispute. This paper points out that the idea implicitly assumes the existence of an overriding authority, e.g., the Courts, to implement the solution. When such an overriding authority does not exist, a different rationale must be employed to make the single‐owner apparatus work. This insight is explored and then applied to help resolve the Taiwan Strait Conflict. Specifically, a ‘One China, Two Republics’ policy is proposed and its implications illustrated.
Public organizations, stakeholders and the construction of publicness. Claims and defence of authority in public action
Authors: Demortain David
Published: December 2004

This article seeks to highlight the conditions in which organizations with different levels of ‘publicness’ interact in the framework of policy‐making. In France, decision making and implementation is shared among public, semi‐public and private organizations. The specific responsibility and influence of public organizations within institutional collective action however is a source of ambiguity. It becomes therefore a matter of political authority for public organizations to invest in the structuring of public problems and policies. The case analysed here shows that the publicness of implemented policies is a construction which stakeholders may try to influence. Responses by public organizations show that publicness does not only relate to the plurality of organizational forms but also to the existence of a form of hierarchy within systems of governance themselves.
Examining the Impact of Workplace Supports: Work‐family Fit and Satisfaction in the U.S. Military
Authors: McFadyen JenniferM. , Kerpelman JenniferL. , Adler‐Baeder Francesca
Published: December 2004

Abstract: The current study sought to discover whether workplace support provided by Army Family Team Building (AFTB) of the Department of the Army was associated with changes in individual knowledge of basic Army lifestyle information, and whether such changes influenced a sense of fit and satisfaction with the Army. Data were collected from 69 Army wives. Findings support the notion that AFTB does increase a basic level of knowledge about the Army, and that this increase positively influences fit. Fit also mediated the relationship between knowledge gain and satisfaction. Implications for the general work‐family interface are discussed.
Factors associated with the effective inclusion of primary‐aged pupils with Down's syndrome
Authors: Fox Sam , Farrell Peter , Davis Pauline
Published: December 2004

It is increasingly common for pupils with Down's syndrome to be offered places in mainstream primary schools. However, there is still much to learn about the impact of these placements upon children and classrooms. Recently the Nuffield Foundation funded a two‐year research project exploring the inclusion of primary‐aged pupils with Down's syndrome. The research focused on the inclusion of 18 pupils with Down's syndrome who attended mainstream primary schools in six local education authorities (LEAs) in the north‐west of England. The research team investigated the ways in which schools manage the inclusion of pupils with Down's syndrome and the factors that contribute to the success of mainstream placements. In this article, Sam Fox, Peter Farrell and Pauline Davis from the University of Manchester discuss some of the findings from the study. These concern the support in place for the child and the attitudes of staff, other pupils and the parents of peers towards the inclusion of the child with Down's syndrome. Evidence from the 18 case studies suggests that there is no single way to guarantee effective inclusion. The extent to which a child is effectively included in a mainstream class is likely to be influenced by a number of key factors, including the way in which the teaching assistant (TA) works with the class teacher. Inclusion is more likely to be successful when the class teacher takes a central role in the management of support and the organisation of a child's daily educational experiences.
Support teachers' beliefs about the academic achievement of students with special educational needs
Authors: Silva José Castro , Morgado José
Published: December 2004

José Castro Silva, lecturer in sciences of education, and José Morgado, assistant professor, both work at the Instituto Superior de Psicologica Aplicada in Lisboa, Portugal. In this article, they describe their study of support teachers’ beliefs about the academic achievement of school students with special educational needs. The ‘support teachers’ who were the subject of this study work in mainstream schools where the majority of pupils with special educational needs are educated in mainstream classes run by ‘general teachers’. The work of the support teachers is supervised and supported by ‘special education team co‐ordinators’. The study reported here set out to elicit the support teachers’ beliefs about the factors that contribute to success at school for pupils with special educational needs. Results suggest that the support teachers consider that factors including ‘school climate’, ‘curriculum design’ and ‘teaching approach’ contribute significantly to achievements among these pupils. On the other hand, analysis reveals that the support teachers attribute difficulties and lack of achievement significantly to ‘out‐of‐school’ contextual variables. These findings are related to a detailed review of the literature and the authors discuss the implications for policy, practice and professional development.
Population pharmacokinetics of epinastine, a histamine H1 receptor antagonist, in adults and children
Authors: Sarashina A. , Tatami S. , Yamamura N. , Tsuda Y. , Igarashi T.
Published: December 2004

Aims

Our objective was to develop a population pharmacokinetic (PPK) model for epinastine, a histamine H1 receptor antagonist, in adults and children and to obtain pharmacokinetic information to support dosing recommendations in children.

Methods

A total of 1510 plasma samples were collected from 62 healthy adult volunteers and 62 paediatric atopic dermatitis patients. The data were analysed using the NONMEM program according to a two‐compartment model with first‐order absorption. In addition, the final PPK model was evaluated by means of bootstrapping resampling.

Results

The oral clearance (CL/F) was found to be associated with body weight, formulation and food status. The volume of distribution of the central compartment (V1/F) was related to body weight and food status. An absorption lag time was apparent in fed subjects. On the other hand, other covariates (formulation on V1/F, volume of distribution of the peripheral compartment (V2/F), first‐order absorption rate constant (Ka) and absorption lag time (ALAG); food status on V2/F and Ka; body weight on V2/F) were not statistically significant. No effect of age on CL/F, V1/F or V2/F was found. The mean parameter estimates obtained with an additional 200 bootstrap replicates of data were within 90–117% of those obtained with the original data set. These results suggest that the pharmacokinetics of epinastine are similar in adults and in children, except for the effect of the difference of body weight. The result of the application of the PPK model to the clinical trial in paediatric patients, in which dosage was determined based on the body weight (from14 kg to less than 24 kg; 10 mg dose, 24 kg or more; 20 mg dose), showed that the Cmax and AUC (25.6 ± 6.9 ng ml−1 and  246.8 ± 68.2 ng h ml−1)  were  almost  same  levels  with those of adults after administration of 20 mg (26.9 ± 9.1 ng ml−1 and 281.6 ± 90.5 ng h ml−1).

Conclusions

A PPK model for epinastine was established and further evaluation by bootstrapping indicated that this model is stable. The model shows that, if dosage is adjusted based on the body weight, the epinastine exposure in paediatric patients is similar to that in adults.
Conditional expression of liver‐enriched transcriptional activator protein augments Acholeplasma laidlawii‐induced granulysin gene expression in a human monocytic cell line, THP‐1
Authors: Kida Yutaka , Shimizu Takashi , Kuwano Koichi
Published: December 2004

The antimicrobial protein granulysin is considered to play an important role in the defence mechanism against bacterial infection. We previously reported that Acholeplasma laidlawii‐induced transactivation of the granulysin promoter in a human monocytic cell line, THP‐1, is regulated by activator protein‐1 and CCAAT/enhancer binding protein‐β (C/EBPβ), but not by nuclear factor‐κB. Moreover, liver‐enriched transcriptional inhibitory protein (LIP), a C/EBPβ isoform, was strongly induced in A. laidlawii‐stimulated THP‐1 cells. However, the level of liver‐enriched transcriptional activator protein (LAP), another C/EBPβ isoform, was essentially constant. Accordingly, we speculated that LIP would down‐regulate A. laidlawii‐induced granulysin gene expression in THP‐1 cells. In the present study, we examined whether LAP augments A. laidlawii‐induced granulysin gene expression using conditional LAP‐expressing THP‐1 cells in a tetracycline‐controlled expression system. Our results indicated that conditional expression of LAP augmented A. laidlawii‐induced expression of granulysin mRNA. In addition, the granulysin protein was observed in A. laidlawii‐stimulated, LAP‐expressing THP‐1 cells. Our results suggest that the expression of LAP plays a critical role in the expression of the granulysin gene in macrophages.
Integrase of Mason–Pfizer monkey virus
Authors: Snášel Jan , Krejčík Zdeněk , Jenčová Věra , Rosenberg Ivan , Ruml Tomáš , Alexandratos Jerry , Gustchina Alla , Pichová Iva
Published: December 2004

The gene encoding an integrase of Mason–Pfizer monkey virus (M‐PMV) is located at the 3′‐end of the pol open reading frame. The M‐PMV integrase has not been previously isolated and characterized. We have now cloned, expressed, isolated, and characterized M‐PMV integrase and compared its activities and primary structure with those of HIV‐1 and other retroviral integrases. M‐PMV integrase prefers untranslated 3′‐region‐derived long‐terminal repeat sequences in both the 3′‐processing and the strand transfer activity assays. While the 3′‐processing reaction catalyzed by M‐PMV integrase was significantly increased in the presence of Mn2+ and Co2+ and was readily detectable in the presence of Mg2+ and Ni2+ cations, the strand transfer activity was strictly dependent only on Mn2+. M‐PMV integrase displays more relaxed substrate specificity than HIV‐1 integrase, catalyzing the cleavage and the strand transfer of M‐PMV and HIV‐1 long‐terminal repeat‐derived substrates with similar efficiency. The structure‐based sequence alignment of M‐PMV, HIV‐1, SIV, and ASV integrases predicted critical amino acids and motifs of M‐PMV integrase for metal binding, interaction with nucleic acids, dimerization, protein structure maintenance and function, as well as for binding of human immunodeficiency virus type 1 and Rous avian sarcoma virus integrase inhibitors 5‐CI‐TEP, DHPTPB and Y‐3.
Structural and functional analysis of the interaction of the AAA‐peroxins Pex1p and Pex6p
Authors: Birschmann Ingvild , Rosenkranz Katja , Erdmann Ralf , Kunau Wolf‐H
Published: December 2004

The AAA‐peroxins Pex1p and Pex6p play a critical role in peroxisome biogenesis but their precise function remains to be established. These two peroxins consist of three distinct regions (N, D1, D2), two of which (D1, D2) contain a conserved ≈ 230 amino acid cassette, which is common to all ATPases associated with various cellular activities (AAA). Here we show that Pex1p and Pex6p from Saccharomyces cerevisiae do interact in vivo. We assigned their corresponding binding sites and elucidated the importance of ATP‐binding and ‐hydrolysis of Pex1p and Pex6p for their interaction. We show that the interaction of Pex1p and Pex6p involves their first AAA‐cassettes and demonstrate that ATP‐binding but not ATP‐hydrolysis in the second AAA‐cassette (D2) of Pex1p is required for the Pex1p–Pex6p interaction. Furthermore, we could prove that the second AAA‐cassettes (D2) of both Pex1p and Pex6p were essential for peroxisomal biogenesis and thus probably comprise the overall activity of the proteins.
Selective modulation of protein C affinity for EPCR and phospholipids by Gla domain mutation
Authors: Preston Roger J. S. , Villegas‐Mendez Ana , Sun Yong‐Hui , Hermida José , Simioni Paolo , Philippou Helen , Dahlbäck Björn , Lane David A.
Published: December 2004

Uniquely amongst vitamin K‐dependent coagulation proteins, protein C interacts via its Gla domain both with a receptor, the endothelial cell protein C receptor (EPCR), and with phospholipids. We have studied naturally occurring and recombinant protein C Gla domain variants for soluble (s)EPCR binding, cell surface activation to activated protein C (APC) by the thrombin–thrombomodulin complex, and phospholipid dependent factor Va (FVa) inactivation by APC, to establish if these functions are concordant. Wild‐type protein C binding to sEPCR was characterized with surface plasmon resonance to have an association rate constant of 5.23 × 105 m−1·s−1, a dissociation rate constant of 7.61 × 10−2 s−1 and equilibrium binding constant (KD) of 147 nm. It was activated by thrombin over endothelial cells with a Km of 213 nm and once activated to APC, rapidly inactivated FVa. Each of these interactions was dramatically reduced for variants causing gross Gla domain misfolding (R‐1L, R‐1C, E16D and E26K). Recombinant variants Q32A, V34A and D35A had essentially normal functions. However, R9H and H10Q/S11G/S12N/D23S/Q32E/N33D/H44Y (QGNSEDY) variants had slightly reduced (< twofold) binding to sEPCR, arising from an increased rate of dissociation, and increased Km (358 nm for QGNSEDY) for endothelial cell surface activation by thrombin. Interestingly, these variants had greatly reduced (R9H) or greatly enhanced (QGNSEDY) ability to inactivate FVa. Therefore, protein C binding to sEPCR and phospholipids is broadly dependent on correct Gla domain folding, but can be selectively influenced by judicious mutation.
Site‐directed mutagenesis and footprinting analysis of the interaction of the sunflower KNOX protein HAKN1 with DNA
Authors: Tioni Mariana F. , Viola Ivana L. , Chan Raquel L. , Gonzalez Daniel H.
Published: December 2004

The interaction of the homeodomain of the sunflower KNOX protein HAKN1 with DNA was studied by site‐directed mutagenesis, hydroxyl radical footprinting and missing nucleoside experiments. Binding of HAKN1 to different oligonucleotides indicated that HAKN1 prefers the sequence TGACA (TGTCA), with changes within the GAC core more profoundly affecting the interaction. Footprinting and missing nucleoside experiments using hydroxyl radical cleavage of DNA showed that HAKN1 interacts with a 6‐bp region of the strand carrying the GAC core, covering the core and nucleotides towards the 3′ end. On the other strand, protection was observed along an 8‐bp region, comprising two additional nucleotides complementary to those preceding the core. Changes in the residue present at position 50 produced proteins with different specificities. An I50S mutant showed a preference for TGACT, while the presence of lysine shifted the preference to TGACC, suggesting that residue 50 interacts with nucleotide(s) 3′ to GAC. Mutation of Lys54→Val produced a protein with reduced affinity and relaxed specificity, able to recognize the sequence TGAAA, while the conservative change of Arg55→Lys completely abolished binding to DNA. Based on these results, we propose a model for the interaction of HAKN1 with DNA in which helix III of the homeodomain accommodates along the major groove with Arg55, Asn51, Lys54 and Ile50, establishing specific contacts with bases of the GACA sequence or their complements. This model can be extended to other KNOX proteins given the conservation of these amino acids in all members of the family.
Role of arterial blood gas abnormalities in oedema formation in COPD
Authors: KARADAG Fisun , POLATLI Mehmet , OZCAN Hatice , CILDAG Orhan
Published: December 2004

Objective:  Renal and hormonal abnormalities, manifesting as oedema or hyponatraemia, are often seen in patients with COPD. The aim of this study was to investigate the effect of airflow obstruction and arterial blood gas abnormalities on oedema formation in COPD patients.

Methodology:  A total of 58 COPD patients hospitalized for treatment of COPD exacerbation were admitted to the study. Of these, 38 patients had peripheral oedema (group 1) and 20 patients had no oedema (group 2).

Results:  The mean age was 68 ± 9 years in group 1 and 68 ± 8 years in group 2. On the first day of admission, serum urea was 29.18 ± 12.25 mg/dL and creatinine was 1.62 ± 0.46 mg/dL in group 1, while urea was 15.50 ± 4.59 mg/dL and creatinine was 1.07 ± 0.10 mg/dL in group 2. Hyponatraemia occurred in five patients (13%) in group 1 and one patient (5%) in group 2. There was no difference in severity of airflow obstruction in the two groups; FEV1 was 44 ± 15% of predicted and FEV1/FVC was 53 ± 14 in group 1, while FEV1 was 45 ± 16% of predicted and FEV1/FVC was 54 ± 20 in group 2. There were statistically significant differences in pH (7.32 vs. 7.39; P= 0.013) and in PaCO2 (62 ± 10 mmHg vs. 42 ± 6; P= 0.048) for group 1 compared with group 2. PaO2 (62 ± 17 mmHg vs. 82 ± 27) and Sao2 (87 ± 9%vs. 90 ± 13) were found to be lower in group 1 compared with group 2 but the difference did not reach statistical significance.

Conclusion:  Alterations in pH and PaCO2 (respiratory acidosis and hypercapnia) appear to have more prominent roles than hypoxaemia in oedema formation in COPD patients.
Adverse effects associated with influenza vaccination in patients with COPD: a randomized controlled study
Authors: WONGSURAKIAT Phunsup , MARANETRA Khun Nanta , GULPRASUTDILOG Penpun , AKSORNINT Monchai , SRILUM Wichian , RUENGJAM Chairat , SATED Wichean
Published: December 2004

Objective:  The aim of this study was to assess the frequency and type of adverse reactions following influenza vaccination and its effects on lung function, dyspnoeic symptoms, exercise capacity, and clinical acute respiratory illness (ARI) in patients with COPD, and the relationship of these adverse effects to the degree of airflow obstruction.

Methodology:  A stratified, randomized, double‐blind placebo‐controlled study was conducted over an 18‐month period at a single university hospital. In total, 125 patients with COPD were randomized to the vaccine group (62 patients who received purified trivalent split‐virus vaccine injections) or the placebo group (63 patients). Local and systemic symptoms during the week following the injections were evaluated. Clinical ARI, lung function tests, dyspnoeic symptoms (assessed using a visual analogue scale), and a 6‐min walking test were evaluated before and at 1 week and 4 weeks following vaccination.

Results:  The frequency of local adverse reactions was 27% in the vaccine group and 6% in the placebo group (P = 0.002). There was no significant difference in systemic adverse reactions between the vaccine and placebo groups (76% vs. 81%; P= 0.5). No difference was observed in the incidence of ARI between the vaccine and placebo groups during the first week (6.4% vs. 6.3%; P= 1) and the first 4 weeks (24.2% vs. 31.7%; P= 0.5) following vaccination. There was no significant change in lung function, dyspnoeic symptoms, and exercise capacity of the patients in both groups, at 1 week and 4 weeks following vaccination, regardless of the severity of COPD.

Conclusion:  Influenza vaccination is associated with minimal local adverse reactions in patients with COPD. Vaccination does not cause systemic adverse reactions, induce clinical exacerbations or adversely affect lung function, dyspnoeic symptoms and exercise capacity in patients with COPD, regardless of the severity of airflow obstruction.
Cost impact of COPD in Japan: Opportunities and challenges?
Authors: NISHIMURA Shuzo , ZAHER Carol
Published: December 2004

Objective:  The Global Initiative for Obstructive Lung Disease highlights the importance of COPD from public health, health policy and clinical perspectives. In countries such as the USA, the economic impact of COPD exceeds that of many chronic conditions. There is a paucity of data on the economic burden of COPD in Japan.

Methodology:  Based upon publicly available information, a prevalence‐based approach was used to construct a deterministic model to estimate the total direct and indirect costs of care for COPD in Japan. Data sources included a spirometry‐based epidemiological study, the peer‐reviewed literature, and governmental and industrial surveys. The most current data that addressed direct and indirect costs of care were utilized.

Results:  In Japan, the estimated total cost of COPD is ¥805.5 billion (US$6.8 billion) per year; ¥645.1 billion (US$5.5 billion) in direct costs and ¥160.4 billion (US$1.4 billion) in indirect costs. In direct costs, inpatient care accounted for ¥244.1 billion (US$2.1 billion), outpatient care ¥299.3 billion (US$2.5 billion), and home oxygen therapy ¥101.7 billion (US$0.9 billion). The average annual total cost per patient for moderate/severe COPD is estimated to be ¥435 876 (US$3694); ¥349 080 (US$2958) per COPD patient in direct costs and ¥86 797 (US$795) in indirect costs.

Conclusion:  COPD imposes a high economic burden on the Japanese healthcare system. Health policy makers should direct urgent attention to increasing prevention, early diagnosis, and appropriate treatment of COPD.
Effects of different anti‐asthmatic agents on induced sputum and eosinophil cationic protein in mild asthmatics
Authors: BASYIGIT Ilknur , YILDIZ Fusun , KACAR OZKARA Sevgiye , BOYACI Hasim , ILGAZLI Ahmet , OZKARAKAS Oktay
Published: December 2004

Objectives:  Inhaled corticosteroids, leukotriene receptor antagonists, and theophylline are recommended for the treatment of mild persistent asthma. The aim of this study was to compare the changes in sputum total cell and eosinophil counts, and eosinophil cationic protein (ECP) levels in serum and sputum following treatment with leukotriene receptor antagonists, inhaled corticosteroids, and theophylline in patients with mild persistent asthma.

Methodology:  Total cell counts, eosinophil percentage, and ECP levels in induced sputum and serum were determined both before and after treatment. Prior to sputum induction, FEV1 and PEF values and symptom scores were recorded at baseline and after 8 weeks of treatment. After baseline measurements, the asthmatic patients (n = 30) were randomized into three groups. A total of 10 patients were treated with zafirlukast, 20 mg bd, 10 with budesonide inhaler 200 µg bd, and 10 with theophylline 200 mg bd.

Results:  There were significant decreases in sputum total cell counts and eosinophil percentage in all treatment groups. However, the decrease in sputum eosinophil counts was more significant in the corticosteroid‐treated group. Although sputum ECP levels decreased significantly in the groups treated with zafirlukast and budesonide (zafirlukast group, 580–135 µg/L, P < 0.01; budesonide group, 683–268 µg/L, P < 0.01), the decrease was not statistically significant in the theophylline‐treated group (498–361 µg/L, P > 0.05). In contrast, there were no significant changes in serum ECP levels in any of the treatment groups.

Conclusions:  All three treatments resulted in significant decreases in sputum total cell counts and eosinophil percentage, but the decrease in sputum ECP level was only seen in the groups treated with budesonide and zafirlukast. These results suggest that although all three treatments are considered as first‐line treatments in most consensuses, theophylline seems to have less of an inhibitory effect on eosinophil activation.
Pathogenesis of pleural fibrosis
Authors: MUTSAERS Steven E. , PRELE Cecilia M. , BRODY Arnold R. , IDELL Steven
Published: December 2004

Abstract:  Pleural fibrosis resembles fibrosis in other tissues and can be defined as an excessive deposition of matrix components that results in the destruction of normal pleural tissue architecture and compromised function. Pleural fibrosis may be the consequence of an organised haemorrhagic effusion, tuberculous effusion, empyema or asbestos‐related pleurisy and can manifest itself as discrete localised lesions (pleural plaques) or diffuse pleural thickening and fibrosis. Although the pathogenesis is unknown, it is likely that the complex interactions between resident and inflammatory cells, profibrotic mediators and coagulation, and fibrinolytic pathways are integral to pleural remodelling and fibrosis. It is generally considered that the primary target cell for pleural fibrosis is the subpleural fibroblast. However, increasing evidence suggests that mesothelial cells may also play a significant role in the pathogenesis of this condition, both by initiating inflammatory responses and producing matrix components. A greater understanding of the interactions between pleural and inflammatory cells, cytokines and growth factors, and blood derived proteins is required before adequate therapies can be developed to prevent pleural fibrosis from occurring.
Management of ischaemic heart disease in women of child‐bearing age
Authors: Wilson A. M. , Boyle A. J. , Fox P.
Published: December 2004

Ischaemic heart disease is rare in young women but is expected to increase with increasing average age of child bearing. Diagnosis of myocardial ischaemia in this group is complicated by limited data about maternal and fetal safety of the standard diagnostic tests routinely used in other patients. Management of these patients remains difficult, as many standard treatments, such as beta‐blockers and angiotensin converting enzyme inhibitors are pregnancy category C or D, and there is little experience with many of the newer treatments such as coronary artery stenting, clopidogrel and glycoprotein IIb/IIIa inhibitors in pregnancy. An interesting case of a woman, who had an acute myocardial infarction treated with thrombolysis and coronary artery stenting, and who subsequently became pregnant, is reported here, and other published reports regarding the management of coronary artery disease, both acute and chronic, in pregnant women are explored. (Intern Med J 2004; 34: 694−697)
The spatial spread of invasions: new developments in theory and evidence
Authors: Hastings Alan , Cuddington Kim , Davies Kendi F. , Dugaw Christopher J. , Elmendorf Sarah , Freestone Amy , Harrison Susan , Holland Matthew , Lambrinos John , Malvadkar Urmila , Melbourne Brett A. , Moore Kara , Taylor Caz , Thomson Diane
Published: December 2004

We review and synthesize recent developments in the study of the spread of invasive species, emphasizing both empirical and theoretical approaches. Recent theoretical work has shown that invasive species spread is a much more complex process than the classical models suggested, as long range dispersal events can have a large influence on the rate of range expansion through time. Empirical work goes even further, emphasizing the role of spatial heterogeneity, temporal variability, other species, and evolution. As in some of the classic work on spread, the study of range expansion of invasive species provides unique opportunities to use differences between theory and data to determine the important underlying processes that control spread rates.
A synthesis of subdisciplines: predator–prey interactions, and biodiversity and ecosystem functioning
Authors: Ives Anthony R. , Cardinale Bradley J. , Snyder William E.
Published: December 2004

The last 15 years has seen parallel surges of interest in two research areas that have rarely intersected: biodiversity and ecosystem functioning (BEF), and multispecies predator–prey interactions (PPI). Research addressing role of biodiversity in ecosystem functioning has focused primarily on single trophic‐level systems, emphasizing additive effects of diversity that manifest through resource partitioning and the sampling effect. Conversely, research addressing predator–prey interactions has focused on two trophic‐level systems, emphasizing indirect and non‐additive interactions among species. Here, we use a suite of consumer‐resource models to organize and synthesize the ways in which consumer species diversity affects the densities of both resources and consumer species. Specifically, we consider sampling effects, resource partitioning, indirect effects caused by intraguild interactions and non‐additive effects. We show that the relationship between consumer diversity and the density of resources and consumer species are broadly similar for systems with one vs. two trophic levels, and that indirect and non‐additive interactions generally do little more than modify the impacts of diversity established by the sampling effect and resource partitioning. The broad similarities between systems with one vs. two trophic levels argue for greater communication between researchers studying BEF, and researchers studying multispecies PPI.
Why environmental scientists are becoming Bayesians
Authors: Clark James S.
Published: December 2004

Advances in computational statistics provide a general framework for the high‐dimensional models typically needed for ecological inference and prediction. Hierarchical Bayes (HB) represents a modelling structure with capacity to exploit diverse sources of information, to accommodate influences that are unknown (or unknowable), and to draw inference on large numbers of latent variables and parameters that describe complex relationships. Here I summarize the structure of HB and provide examples for common spatiotemporal problems. The flexible framework means that parameters, variables and latent variables can represent broader classes of model elements than are treated in traditional models. Inference and prediction depend on two types of stochasticity, including (1)uncertainty, which describes our knowledge of fixed quantities, it applies to all ‘unobservables’ (latent variables and parameters), and it declines asymptotically with sample size, and (2)variability, which applies to fluctuations that are not explained by deterministic processes and does not decline asymptotically with sample size. Examples demonstrate how different sources of stochasticity impact inference and prediction and how allowance for stochastic influences can guide research.
The Case for Small Area Microdata
Authors: Tranmer Mark , Pickles Andrew , Fieldhouse Ed , Elliot Mark , Dale Angela , Brown Mark , Martin David , Steel David , Gardiner Chris
Published: December 2004

Census data are available in aggregate form for local areas and, through the samples of anonymized records (SARs), as samples of microdata for households and individuals. In 1991 there were two SAR files: a household file and an individual file. These have a high degree of detail on the census variables but little geographical detail, a situation that will be exacerbated for the 2001 SAR owing to the loss of district level geography on the individual SAR. The paper puts forward the case for an additional sample of microdata, also drawn from the census, that has much greater geographical detail. Small area microdata (SAM) are individual level records with local area identifiers and, to maintain confidentiality, reduced detail on the census variables. Population data from seven local authorities, including rural and urban areas, are used to define prototype samples of SAM. The rationale for SAM is given, with examples that demonstrate the role of local area information in the analysis of census data. Since there is a trade-off between the extent of local detail and the extent of detail on variables that can be made available, the confidentiality risk of SAM is assessed empirically. An indicative specification of the SAM is given, having taken into account the results of the confidentiality analysis.