bluuebunny/crossref_metadata_2025 huggingface dataset:
White‐core endosperm floury endosperm‐4 in rice is generated by knockout mutations in the C4‐type pyruvate orthophosphate dikinase gene (OsPPDKB)
Authors: Kang Hong‐Gyu , Park Sunhee , Matsuoka Makoto , An Gynheung
Published: June 2005

We have isolated a floury endosperm‐4 (flo4) rice mutant with a floury‐white endosperm but a normal outer portion. Scanning electron microscopic analysis revealed that this abnormal endosperm consisted of loosely packed starch granules. The mutant phenotype was generated by T‐DNA insertion into the fifth intron of the OsPPDKB gene encoding pyruvate orthophosphate dikinase (PPDK). Plants containing flo4‐1 produced no OsPPDKB transcript or the OsPPDKB protein in their developing kernels and leaves. We obtained two additional alleles, flo4‐2 and flo4‐3, that also showed the same white‐core endosperm phenotype. The flo4 kernels weighed about 6% less than wild‐type ones. Starch contents in both kernel types were similar, but the total protein content was slightly higher in the mutant kernels. Moreover, lipid contents were significantly increased in the flo4 kernels. Expression analyses demonstrated that the cytosolic mRNA of OsPPDKB was induced in the reproductive organs after pollination, and greatly increased until about 10 days after fertilization. This mRNA was localized mainly in the endosperm, aleurone, and scutellum of the developing kernel. Our results suggest that cytosolic PPDK functions in rice to modulate carbon metabolism during grain filling.
The eukaryotic translation initiation factor 4E confers multiallelic recessive Bymovirus resistance in Hordeum vulgare (L.)
Authors: Stein Nils , Perovic Dragan , Kumlehn Jochen , Pellio Bettina , Stracke Silke , Streng Stefan , Ordon Frank , Graner Andreas
Published: June 2005

Virus diseases are widespread threats for crop production, which can, in many cases, be controlled efficiently by exploiting naturally occurring resistance. Barley, an important cereal species of the Triticeae, carries two genes, rym4 and rym5, which are located in the telomeric region of chromosome 3HL and confer recessive resistance to various strains of the Barley yellow mosaic virus complex. The barley ‘eukaryotic translation initiation factor 4E’ (Hv‐eIF4E) was identified as a candidate for resistance gene function by physical mapping on a 650 kb contig. It is located in a chromosomal region characterized by suppressed recombination, in a position collinear to its homologue on rice chromosome 1L. Sequence diversity in the coding region of Hv‐eIF4E, as calculated from a collection of unrelated barley accessions, revealed non‐silent single nucleotide polymorphisms (SNPs) in four of its five exons. Stable transformation of a resistant barley genotype with a genomic fragment or a full‐length cDNA of Hv‐eIF4E derived from susceptible cultivars induced susceptibility to Barley mild mosaic virus. Moreover, the identification of SNPs diagnostic for rym4 and rym5 provides evidence that these are two alleles, which confer different resistance specificities. These findings demonstrate that variants of Hv‐eIF4E confer multiallelic recessive virus resistance in a monocot species. The identification of eIF4E as the causal host factor for bymovirus resistance illustrates that mutations in this basic component of the eukaryotic translation complex form a seminal mechanism for recessive virus resistance in both dicot and monocot plants.
The effect of encasings on quality of life in adult house dust mite allergic patients with rhinitis, asthma and/or atopic dermatitis
Authors: Terreehorst I. , Duivenvoorden H. J. , Tempels‐Pavlica Z. , Oosting A. J. , de Monchy J. G. R. , Bruijnzeel‐Koomen C. A. F. M. , van Wijk R. Gerth
Published: June 2005

Background:  Environmental control has been put forward as an integral part of the management of house dust mite (HDM) allergy in sensitized patients. To validate this statement allergic disorders involved in HDM allergy – allergic asthma, rhinitis and atopic eczema/dermatitis syndrome (AEDS) – should be taken together and studied in terms of the efficacy of environmental control. Because a generic quality of life questionnaire exceeds the border of disease, this may be used as major outcome parameter.

Research objective:  To study the effects of bedding encasings in HDM allergic patients with asthma, rhinitis and AEDS.

Material and methods:  A total of 224 adult HDM allergic patients with rhinitis and/or asthma and/or dermatitis were randomly allocated impermeable or nonimpermeable encasings for mattress, pillow and duvet. Short form 36 (SF‐36) was filled in at baseline and after 12 months.

Results:  Lower physical (P = 0.01) and emotional (P < 0.001) sumscores were seen in females. Also, the presence of asthma resulted in lower physical sumscore (P = 0.01). However, no effect was seen of encasings on either sumscore.

Conclusion:  Bedding encasings do not improve quality of life in a mixed population of subjects with combinations with rhinitis, asthma and atopic dermatitis and sensitized to HDMs.
Association of interleukin 18 (IL18) polymorphisms with specific IgE levels to mite allergens among asthmatic patients
Authors: Shin H. D. , Kim L. H. , Park B. L. , Choi Y. H. , Park H.‐S. , Hong S.‐J. , Choi B. W. , Lee J. H. , Park C.‐S.
Published: June 2005

Background:  Allergy is regarded as a multifactorial condition. Its onset and severity are influenced by both genetic and environmental factors. Identification of genetic factors involved in asthma development and related phenotypes is a major task in understanding the genetic background of asthma. The possible involvement of IL18 polymorphisms in asthma was examined in a Korean asthma cohort.

Methods:  Direct sequencing was performed to discover single‐nucleotide polymorphisms (SNPs) in the IL18 gene. Single‐base extension (SBE) method was employed for genotyping. Genotypic influence of IL18 was analysed using logistic and multiple‐regression models.

Results:  Although no polymorphisms in the IL18 gene showed significant association with the risk of asthma development, analyses of the association with specific serum IgE levels to Dermatophagoides farinae (D.f.) and D. pteronyssinus (D.p.) among asthmatic patients revealed significant associations with two completely linked SNPs, i.e. −148G>C and +13925A>C(Ser35Ser) (P = 0.01–0.11 for D.f. and P = 0.005–0.11 for D.p.). Both C allele of −148G>C and C allele of +13925A>C showed gene dose‐dependent effects on the levels of specific IgE. The lowest IgE levels in homozygotes of minor alleles (1.13 and 1.22 of D.f.; 1.38 and 1.33 of D.p., respectively), intermediate IgE levels in heterozygotes (1.60 and 1.70 of D.f.; 1.84 and 1.92 of D.p., respectively), and the highest levels in homozygotes for major allele (1.93 and 1.93 of D.f.; 2.24 and 2.24 of D.p., respectively), were found.

Conclusion:  The genetic relevance of IL18 to specific IgE might offer an important step in understanding the genetic background of allergic diseases.
Relationships between allergic inflammation and nasal airflow in children with persistent allergic rhinitis due to mite sensitization
Authors: Ciprandi G. , Marseglia G. L. , Klersy C. , Tosca M. A.
Published: June 2005

Background:  Allergic rhinitis is associated with Th2‐dependent inflammation. Nasal obstruction is the most typical symptom in children with mite allergy.

Objectives:  The aim of this study was to evaluate the possible relationships among nasal symptoms, allergic inflammation, including inflammatory cells and cytokine pattern, and nasal airflow in children with persistent allergic rhinitis because of mite sensitization.

Methods:  Twenty children (13 males and seven females, mean age 13.4 ± 1.6 years) with persistent rhinitis because of mite allergy were evaluated. All of them had moderate–severe grade of nasal obstruction. Total symptom score (TSS), rhinomanometry, nasal lavage, and nasal scraping were obtained in all subjects. Inflammatory cells were counted by conventional staining; interleukin (IL)‐5, and IL‐8 were measured by immunoassay on fluids recovered from nasal lavage.

Results:  Eosinophils were significantly associated with TSS (R = 74.4%, P = 0.0002), with IL‐5 (R = 90.6%, P < 0.0001) and with nasal flow (R = −69%, P = 0.0007), but not with IL‐8 (R = 0.1%, P = 0.995). Eosinophil levels were shown to independently predict nasal flow (P < 0.001), with flow decreasing linearly for increasing eosinophils, together with a significant effect of neutrophils (P = 0.016, linear increase in flow) and a borderline effect of IL‐8 (P = 0.063, linear increase in flow).

Conclusions:  This study demonstrates the close association between IL‐5 concentration and eosinophil infiltration. In addition, there is clear evidence concerning the relationship between eosinophil infiltration and nasal airflow. Thus, nasal eosinophils can be regarded as the most important predictor of upper airway function. These findings constitute first evidence of the relationship between nasal airflow impairment and Th2‐related eosinophilic inflammation in children with persistent allergic rhinitis because of mite sensitization.
Dose dependence of immunological response to sublingual immunotherapy
Authors: Marcucci F. , Sensi L. , Di Cara G. , Incorvaia C. , Frati F.
Published: June 2005

Background:  Sublingual‐swallow immunotherapy (SLIT) is an accepted treatment for allergic rhinitis but its optimal dosage is scantly investigated. We studied the dose dependence of clinical efficacy and immunological response to SLIT by administering two different dosages of the same allergen in rhinitic children monosensitized to grass pollen.

Methods:  Seventy‐one patients with comparable age and symptoms were randomized to receive SLIT by the same grass pollen extract from Stallergénes (Antony, France), 40 of them with the 100 IR and 31 with the 300 IR extract. All patients recorded diary cards for symptoms, medications and side‐effects of the treatment, and had measurements of specific IgE and IgG4 in serum by the CAP System FEIA (Pharmacia, Uppsala, Sweden) and in nasal secretion by an in situ incubation method with the same reagents of CAP System FEIA.

Results:  Symptom/medication scores during the pollen season were significantly higher in patients treated with the lower dosage compared with those treated with the 300 IR dosage. Side‐effects occurred with a comparable rate (25.8%vs 27.5%) in the two groups. Serum‐specific IgE and IgG4 had no significant changes after 3 months of SLIT in both groups, while a significant seasonal increase of nasal IgE (P = 0.015) and IgG4 (P = 0.019) was found only in patients treated with the lower dosage.

Conclusions:  A rise of specific IgG4 and a blunting of seasonal increase of specific IgE in serum was repeatedly reported during subcutaneous immunotherapy (SCIT) with pollen extracts. Our findings show such blunting of specific nasal IgE along with a low symptom/medication score in patients treated with SLIT with the higher dosage, but not a concomitant rise of specific nasal IgG4. This suggests a local immunological effect of SLIT, different from systemic mechanisms of SCIT.
Diverging prevalence trends of atopic disorders in Norwegian children. Results from three cross‐sectional studies
Authors: Selnes A. , Nystad W. , Bolle R. , Lund E.
Published: June 2005

Background:  During the last decades there has been extensive epidemiological research to explore the increasing prevalence of asthma and allergy in childhood. The worldwide variations in prevalence of these diseases necessitate regional rapports. Furthermore, time‐trend analyses with comparable methods are important in order to monitor the rapidly changing prevalence of these diseases.

Methods:  Three cross‐sectional questionnaire‐based studies of asthma and allergy in schoolchildren were conducted in the counties of Troms and Finnmark, in northern Norway in 1985, 1995 and 2000. The two former studies included children from randomly selected primary schools (n = 1794/1985, n = 1432/1995). The latter study was a part of ISAAC‐II Europe study (n = 3853). Identical items of asthma and allergy were employed. The analyses comprised only children 9–11 years of age.

Results:  The prevalence of asthma was 9.3, 13.2 and 13.8% in 1985, 1995 and 2000, respectively. However, great gender differences were detected; the prevalence of asthma increased in males from 1995 to 2000, from 14.1 to 17.0%, RR = 1.2 (95% CI 1.0–1.5), but decreased in females 1995 to 2000, from 12.3 to 10.5%, RR = 0.9 (95% CI 0.7–1.1). Furthermore, in children with asthma, a changing trend was found in the external factors that perceived symptoms, from typical allergens towards other, unspecific agents. The prevalence of self‐reported atopic eczema/dermatitis syndrome (AEDS) was 13.4, 21.1 and 20.8% in 1985, 1995 and 2000, respectively. The prevalence of self‐reported allergic rhinoconjunctivitis was in 16.5, 24.7 and 29.6% 1985, 1995 and 2000, respectively, RR (2000/1995) = 1.2 (95% CI 1.1–1.3).

Conclusion:  The prevalence of asthma in girls has reached a plateau and even decreased from 1995 to 2000 which is in contrast to the asthma prevalence in boys that tends to continuously increase. The prevalence of AEDS which increased substantially between 1985 and 1995 did not change from 1995 to 2000. However, the prevalence of allergic rhinoconjunctivitis increased steadily from 1985, 1995 to 2000.
Inflammatory cell numbers and cytokine expression in atopic dermatitis after topical pimecrolimus treatment
Authors: Simon D. , Vassina E. , Yousefi S. , Braathen L. R. , Simon H.‐U.
Published: June 2005

Background:  In several clinical trials the topical application of pimecrolimus was shown to be effective in the treatment of atopic dermatitis (AD). By targeting calcineurin‐dependent signaling pathways, pimecrolimus controls cytokine gene expression. The purpose of this study was to investigate the effect of pimecrolimus on the inflammatory infiltrate and cytokine expression pattern in AD upon topical therapy.

Methods:  From 10 patients with acute AD, skin biopsies as well as immunophenotype and cytokine production of peripheral blood mononuclear cells (PBMC) were examined before and 3 weeks after therapy.

Results:  The clinical improvement was associated with a marked regression of histopathological features. In particular, the density of the inflammatory infiltrate mostly containing lymphocytes and eosinophils declined. By double immunofluorescent staining, a reduced expression of the T helper (Th) 2 cytokines interleukin (IL)‐5, IL‐10, and IL‐13 in both CD4+ and CD8+ T cells was demonstrated after therapy. Pimecrolimus therapy was also associated with a reduced expression of the Th1 cytokine interferon (IFN)‐γ. Interestingly, the numbers of epidermal CD1a+ dendritic cells increased following treatment. In the peripheral blood, a decrease of lymphocytes and eosinophils was noticed, but the distribution of lymphocyte subpopulations and their capacity of cytokine production did not change.

Conclusions:  Topical pimecrolimus exhibits anti‐inflammatory effects in AD by reducing the inflammatory cell infiltrate and cytokine expression in the dermis.
Eosinophilic pneumonias
Authors: Cottin V. , Cordier J.‐F.
Published: June 2005

Eosinophilic pneumonias (EP) encompass a wide spectrum of lung diseases characterized by peripheral blood eosinophilia (>1 × 109 eosinophils/l) and/or alveolar eosinophilia (>25%). Blood eosinophilia may be lacking, as in the early phase of idiopathic acute EP, or in patients already taking oral corticosteroids. EP may present with varying severity, ranging from almost asymptomatic infiltrates to the acute respiratory distress syndrome necessitating mechanical ventilation. Possible causes of EP must be thoroughly investigated, especially drugs and the variety of parasitic infections (considering history of travel or residence in areas of endemic parasitic infection). However, chronic EP remains idiopathic in many cases. When present, extrathoracic manifestations lead to suspect Churg–Strauss syndrome (CSS) or the hypereosinophilic syndrome (HES), the prognosis of which is dominated by cardiac involvement. Apart from the treatment of specific causes when possible, corticosteroids remain the cornerstone of symptomatic treatment for eosinophilic disorders, usually with a dramatic response, but frequent relapses when tapering or after stopping the treatment. The adjunction of immunosuppressants to corticosteroids is necessary in patients with CSS and poor prognosis factors. Imatinib has recently proven effective in the treatment of the myeloproliferative variant of the HES.
Sub‐lethal exposure of cockroaches to boric acid pesticide contributes to increased Bla g 2 excretion
Authors: Zhang Y. C. , Perzanowski M. S. , Chew G. L.
Published: June 2005

Background:  Several epidemiology studies have found an increase in the major cockroach allergen Bla g 2 with reported pesticide use.

Aims of the study:  Our aim was to investigate the effect on the excretion of Bla g 1 and Bla g 2 allergens by cockroaches exposed to sub‐lethal doses of the pesticides, boric acid and hydramethylnon gel.

Methods:  German cockroaches in separate colonies were fed either boric acid or hydramethylnon gel at concentrations of 0.2, 0.1 and 0.01% in their water supply over a 2 week period. Ten colonies were exposed to each treatment concentration. Bla g 1 and Bla g 2 in fecal pellets were measured by ELISA.

Results:  Cockroaches exposed to boric acid excreted fecal pellets with significantly higher concentrations of Bla g 2 (35 400 U/g) than did controls (12 700 U/g) (P = 0.001). Bla g 1 concentrations were not significantly different. There was no difference in either Bla g 1 or Bla g 2 concentrations between cockroaches that ingested hydramethylnon gel and those in the controls colonies.

Conclusions:  The application of boric acid, a common pesticide, appears to paradoxically increase the production of Bla g 2, a major allergen, by the surviving cockroaches. This may have important implications in avoidance strategies.
The effect of interference‐fit fasteners on the fatigue life of central hole specimens
Authors: LANCIOTTI A. , POLESE C.
Published: June 2005

Fatigue tests were carried out on 2024‐T351, thickness 1.6 mm, central hole specimens containing pins installed with five different interference‐fit levels. Tests clearly demonstrated the beneficial effect of interference fit on fatigue resistance, up to the maximum value examined, 2.5%. A three‐dimensional (3D) finite‐element model was used in order to characterize the stress field around the hole. A large specimen, with a 40‐mm‐diameter hole filled with interference‐fit pin, was instrumented by strain gauges and statically tested in order to check FEM results. A very good correlation existed between measured and numerically evaluated strains. FEM results demonstrated the well‐known effect of interference‐fit fasteners on reducing stress ranges. By increasing the interference level, the stress range was practically unchanged, while the mean stress decreased. Interference‐fit produces a biaxial stress state, which must be taken into account for fatigue evaluation. In the present case, a simple criterion, based on hoop strain, predicted the fatigue results quite well with the exception of open hole fatigue test results, which were overestimated.
Standing contact fatigue with a cylindrical indenter
Authors: DAHLBERG J. , ALFREDSSON B.
Published: June 2005

A hardened steel cylinder was repeatedly pressed against a flat case‐hardened steel specimen that was equally wide as the cylinder was long. Some contact end effects were noted as a result of limited plastic deformation. A strain gauge on the contact surface, just outside the contact and oriented perpendicular to the cylinder detected a surface strain when the cylinder was loaded. The non‐zero surface strain was the result of boundary effects of the finite specimen. Four different types of contact fatigue cracks developed in and below the specimen contact surface. The cracks were named lateral, median, contact end and edge cracks. Changes in the measured surface strain values could be used to determine when the lateral and edge cracks developed. The order in which all four crack types typically developed was determined from optical crack observation at test termination, strain measurements and stress computations. Numerical computations using finite‐element (FE) analyses were used to verify the surface strain behaviour due to loading and cracking; to verify contact end effects; crack locations and crack orientation by aid of the Findley multi‐axial fatigue criterion.
Fatigue resistance of CrN thin films deposited by arc evaporation process on H11 tool steel and 2205 duplex stainless steel
Authors: BARAGETTI S. , GELFI M. , LA VECCHIA G.M. , LECIS N.
Published: June 2005

Notwithstanding the wide use of PVD coated components, not only in fields where tribological behaviour is important and well assessed, fatigue behaviour is still an object of research in applications where enhanced fatigue resistance is required. The aim of this paper is to analyse fatigue crack growth mechanisms and to relate their nucleation sites to the microstructure and to residual stress data. Two structural steels were used as the substrate (H11 tool steel grade and 2205 duplex stainless steel grade), while a monolayer of CrN, 5 μm deep, was deposed as a coating. Four‐point bending fatigue tests were carried out in order to obtain the bending fatigue limit. Experimental results obtained by scanning electron microscopy analyses, residual stresses measurements and hardness evaluations carried out using a nanoindenter device were considered to explain the fatigue mechanisms involving a PVD coated part in terms both of nucleation site and crack growth process.
Avoiding knife‐edge countersinks in GLARE through dimpling
Authors: RANS C. , STRAZNICKY P. V.
Published: June 2005

Traditional machine countersinking practices create a knife‐edge condition in one or more of the outer aluminium layers in riveted GLARE joints. Press countersinking (dimpling) provides an alternative method of countersinking that prevents the formation of a knife‐edge; however, its application and potential benefits to fatigue performance in GLARE are not known. This paper investigates the dimple‐forming process and its application to GLARE, and the resulting benefits in fatigue crack‐initiation life in unfilled rivet holes. Initial results showed that the limited formability of GLARE complicates the dimpling process, but that dimpling shows promise as a method for increasing the crack‐initiation life of riveted GLARE joints.
Time‐dependent fracture of type 316L(N) steel at ambient temperature
Authors: BUDDEN P. J. , WARDLE G. , BIRKETT R. P.
Published: June 2005

Experimental data on tensile and compact geometry (CT) specimens of austenitic Type 316L(N) steel were obtained under sustained load conditions at room temperature. Time‐dependent crack growth, in some cases leading to failure, occurred in many of the CT specimens, dependent on the load level. However, rupture of the uniaxial specimens occurred only at stresses very close to the material's ultimate strength. The data validate the approach to assessing sustained loading effects in the R6 defect assessment procedure. In particular, sustained load effects in austenitic steel may be neglected for values of the R6Lrparameter less than unity. Uniaxial sustained load tests were also performed at 100 °C and 200 °C. The measured strain rates decreased with increasing temperature, becoming negligible at 200 °C. This is consistent with the advice in R6 that sustained load effects in austenitic steel can be neglected at temperatures between 200 °C and the high‐temperature creep range.
Complexity Theory and Strategic Change: an Empirically Informed Critique*
Authors: Houchin K. , MacLean D.
Published: June 2005

We present a four‐year ethnographic study of a public‐sector organization and use narrative to describe its development in terms of four complexity theory concepts: sensitivity to initial conditions, negative and positive feedback processes, disequilibrium and emergent order. Our study indicates that order emerges at the boundary between the organization's legitimate and shadow systems. We suggest that the underlying dynamic leading to the emergent order is the need to reduce anxiety. Our findings cause us to question the assertion that organizations are naturally complex adaptive systems producing novel forms of order. We propose an alternate view that in social systems, equilibrium‐seeking behaviour is the norm; such systems can self‐organize into hierarchy. We draw attention to some of the difficulties we found in applying complexity‐theory concepts to a social system and conclude by advocating the development of complexity theory through the incorporation of insights from psychology and social theory.
The Glass Cliff: Evidence that Women are Over‐Represented in Precarious Leadership Positions
Authors: Ryan Michelle K. , Haslam S. Alexander
Published: June 2005

There has been much research and conjecture concerning the barriers women face in trying to climb the corporate ladder, with evidence suggesting that they typically confront a ‘glass ceiling’ while men are more likely to benefit from a ‘glass escalator’. But what happens when women do achieve leadership roles? And what sorts of positions are they given? This paper argues that while women are now achieving more high profile positions, they are more likely than men to find themselves on a ‘glass cliff’, such that their positions are risky or precarious. This hypothesis was investigated in an archival study examining the performance of FTSE 100 companies before and after the appointment of a male or female board member. The study revealed that during a period of overall stock‐market decline those companies who appointed women to their boards were more likely to have experienced consistently bad performance in the preceding five months than those who appointed men. These results expose an additional, largely invisible, hurdle that women need to overcome in the workplace. Implications for the evaluation of women leaders are discussed and directions for future research are outlined.
An Analysis of the Relationship between Environmental Motivations and ISO14001 Certification
Authors: González‐Benito Javier , González‐Benito Oscar
Published: June 2005

The objectives of this paper are both to identify the environmental motivations associated with the decision to initiate the process for ISO14001 certification and to check whether such motivations undergo any changes once the company obtains its certification and experiences the actual effects of this process. Based on the work of Bansal and Roth, we distinguish between ethical, competitive and relational motivations, and compare the importance of these motivations in four company stages with respect to the ISO14001 certification: not considered, considered, in process and awarded. This comparison was performed through a logit analysis applied over the data supplied by 184 Spanish manufacturers from three different industrial sectors. It was concluded that the decision to pursue the ISO14001 certification responds to ethical and competitive motivations, and that once the company gets its certification, the company portfolio of environmental motivations does not change significantly.
Impact of French ‘Comités de Protection des Personnes’ on the readability of informed consent documents (ICD) in biomedical research: more information, but not better information
Authors: Paris Adeline , Cracowski Jean‐Luc , Maison Patrick , Radauceanu Anca , Cornu Catherine , Hommel Marc
Published: June 2005

Information is the keystone to the participation of subjects in biomedical research. Clear comprehension of the informed consent documents (ICDs) is primordial and a necessary requirement is that they are readable. While submission of a protocol to a French ‘Comités de Protection des Personnes’ (CPP) is a mandatory step with regard to the French legislation on biomedical research, no published data are available concerning its influence on ICDs readability. The aim of our study was to determine the impact of French CPP on the readability of ICDs, using lexico‐syntactic readability indexes and ICDs from four clinical research centres and one clinical research unit. Twenty‐five ICDs were analysed. The Flesch score was not modified after CPP review, while the Cordial score was significantly lower [from 4 (1–14) to 1 (1–13), P = 0.014]. The information was longer and more complex following CPP review. No protocol characteristics had any impact on the variation before and after review for either the Flesch or the Cordial® indexes, nor on the number of syllables per word. Changes in the total number of words before and after review varied considerably between study centre, supporting heterogeneity of CPP review. Since August 2004, French CPP have to study the intelligibility of ICDs in addition to the scientific and ethic aspects of a research. We show that their current reviews do not increase the readability, while increasing the length of ICDs.
Cooling and response to hydrogen peroxide in human saphenous vein: role of the endothelium
Authors: Şahin Ayşe Saide , Atalık Kısmet Esra , Şahin Tahir Kemal , Doğan Necdet
Published: June 2005

In the present work we studied the responses of human saphenous vein to H2O2 and effects of moderate cooling on these responses with analysis of the role of endothelium. H2O2 (10−7–10−2 m) induced concentration‐dependent contraction in the intact human saphenous vein strips at both temperatures. At 28 °C, the maximal contraction induced by H2O2 was significantly lower than that at 37 °C. Compared with intact strips, the sensitivity and the maximal contraction to H2O2 were significantly enhanced in endothelium‐denuded strips at 37 and 28 °C. However, pD2 values and maximal contractions were not significantly different in endothelium‐denuded strips at different temperatures. Pretreatment with NG‐nitro‐l‐arginine methyl ester (l‐NAME) increased significantly the maximal contraction and sensitivity to H2O2 at 37 and 28 °C. The contractions increased by l‐NAME were restored by the pre‐incubation of l‐arginine (10−3 m) at every temperature studied. The contractile responses of intact human saphenous veins to H2O2 were reduced significantly by 10−5 m indomethacin at both temperatures. Our results suggest that H2O2‐induced contraction of human saphenous vein are mediated by its direct effect on the smooth muscle and by the generation of products of the cyclooxygenase pathway from the endothelium. Signalling pathways of these contractile effects are the same at 37 and 28 °C. Under normal temperature conditions, the contraction to H2O2 is possibly modulated by endothelial nitric oxide. Cooling reduces the contraction to H2O2 by increasing release of nitric oxide.
Analysis of the AP‐2 Adaptor Complex and Cargo During Clathrin‐Mediated Endocytosis
Authors: Rappoport Joshua Z. , Benmerah Alexandre , Simon Sanford M.
Published: June 2005

Previously, we reported that the hetero‐tetrameric adaptor complex AP‐2 co‐localizes with the static population of clathrin spots, whereas it is excluded from clathrin spots that disappear from the plasma membrane (forming clathrin‐coated vesicles). More recently however, another group provided evidence that AP‐2 markers could be observed coincident with disappearing clathrin spots. Thus, we tested several possible explanations for the apparent discrepancies in these two studies. We evaluated the potential contribution of nonred emission of clathrin‐dsRed (used in both studies) in the simultaneous measurement of AP‐2 and clathrin at various times. Additionally, we directly compared two different green fluorescent protein‐tagged AP‐2 constructs (similar to those used in the previous reports). These studies demonstrated that the duration of expression time greatly influences the subcellular localization of the AP‐2 markers. Furthermore, we quantitatively evaluated the AP‐2 fluorescence at the sites of numerous static and disappearing clathrin spots (at least 80 per group) and confirmed our initial observation that while AP‐2 is present in nearly all static clathrin spots, it is excluded from the disappearing population of clathrin spots. Finally, in order to verify that clathrin spot disappearance represents clathrin‐coated vesicle internalization, we simultaneously imaged clathrin and the cargo molecule transferrin at the cell surface.
Brefeldin A Inhibits Circadian Remodeling of Chloroplast Structure in the Dinoflagellate Gonyaulax
Authors: Nassoury Nasha , Wang Yunling , Morse David
Published: June 2005

Circadian increases in the rate of carbon fixation in the dinoflagellate Gonyaulax are correlated with extensive plastid remodeling. One marker for this remodeling is mobilization of ribulose bisphosphate carboxylase/oxygenase (Rubisco) from the plastid periphery to plastid regions nearer the cell center called pyrenoids. Nuclear‐encoded proteins such as Rubisco transit through the Golgi in dinoflagellates; hence, we blocked protein import into the plastids using Brefeldin A (BFA) to explore the mechanism for plastid remodeling. We find that pyrenoid formation normally occurs concurrently with increased Rubisco synthesis rates in vivo, and when BFA is given prior to the onset of Rubisco synthesis, pyrenoid formation is partially or completely inhibited by 0.1 or 0.3 µg/mL BFA, respectively. Rubisco synthesis itself is not affected, and BFA‐treated cells accumulate Rubisco in novel structures we term BFA bodies. Interestingly, when given just after the onset of Rubisco synthesis, BFA delays but does not block Rubisco mobilization, suggesting that a timing signal for plastid remodeling is delivered to the organelles at the same time as newly synthesized Rubisco. BFA also inhibits the circadian increases in carbon fixation rates, supporting the hypothesis that the biochemical basis for this circadian rhythm may be Rubisco distribution within the plastid.
Endoplasmic Reticulum Exit of a Secretory Glycoprotein in the Absence of Sec24p Family Proteins in Yeast
Authors: Karhinen Leena , Bastos Ricardo Nunes , Jokitalo Eija , Makarow Marja
Published: June 2005

Glycoproteins exit the endoplasmic reticulum (ER) of the yeast Saccharomyces cerevisiae in coàt protein complex II (COPII) coated vesicles. The coat consists of the essential proteins Sec23p, Sec24p, Sec13p, Sec31p, Sar1p and Sec16p. Sec24p and its two nonessential homologues Sfb2p and Sfb3p have been suggested to serve in cargo selection. Using temperature‐sensitive sec24‐1 mutants, we showed previously that a secretory glycoprotein, Hsp150, does not require functional Sec24p for ER exit. Deletion of SFB2, SFB3 or both from wild type or the deletion of SFB2 from sec24‐1 cells did not affect Hsp150 transport. SFB3 deletion has been reported to be lethal in sec24‐1. However, here we constructed a sec24‐1 Δsfb3 and a sec24‐1 Δsfb2 Δsfb3 strain and show that Hsp150 was secreted slowly in both. Turning off the SEC24 gene did not inhibit Hsp150 secretion either, and the lack of SEC24 expression in a Δsfb2 Δsfb3 deletant still allowed some secretion. The sec24‐1 Δsfb2 Δsfb3 mutant grew slower than sec24‐1. The cells were irregularly shaped, budded from random sites and contained proliferated ER at permissive temperature. At restrictive temperature, the ER formed carmellae‐like proliferations. Our data indicate that ER exit may occur in vesicles lacking a full complement of Sec23p/24p and Sec13p/31p, demonstrating diversity in the composition of the COPII coat.
Evaluation of Lead Zirconate–Titanate Ceramic Target for Sputter Deposition of Films for Use in Ferroelectric Capacitors
Authors: Cross Jeffrey Scott , Tsukada Mineharu , Goto Yasuyuki , Kamehara Nobuo , Nagayama Satsuki
Published: June 2005

A high‐density 3 in. diameter (Pb,La)(Zr,Ti)O3 [PLZT] ceramic target containing 20% excess PbO was sputtered by radio‐frequency magnetron sputtering to deposit films containing excess Pb and evaluated with respect to target stability for use in the mass production of ferroelectric memories on Si wafers. The results indicated that PLZT film deposition rate increased with power but the excess PbO content correspondingly decreased. The Pb loss in the films was attributed to secondary preferential sputtering of film substrate in the plasma. Regardless of this difference, excess PbO containing 12 in. diameter PLZT ceramic targets were evaluated in a commercial sputtering system over a period of 7 months and the film Pb content was stable within ±1%.
Green Machining to Net Shape Alumina Ceramics Prepared Using Different Processing Routes
Authors: Dhara Santanu , Su Bo
Published: June 2005

In the present study, the net shape manufacturing of alumina ceramics was carried out using computer numerically controlled (CNC) machining of differently processed alumina green bodies using different tool materials and geometries. In case of a carbide‐coated tool, severe wear was observed; on the contrary, a diamond‐coated tool was used successfully. The profile of the as‐machined surface was found to be smooth using diamond‐coated flat tool. Near net shape alumina was demonstrated using CNC green machining from optimized machining conditions. Optical micrograph and the surface roughness of the as‐machined samples showed good surface finish for the green bodies with higher binder content.
Three‐Dimensional Microstructural Characterization of Porous Hydroxyapatite Using Confocal Laser Scanning Microscopy
Authors: Ren Fei , Smith Ian O. , Baumann Melissa J. , Case Eldon D.
Published: June 2005

The characterization of porosity is crucial in the development and commercialization of ceramic bone replacement technology, since the pore size and interconnectivity play a central role in both biological function (bone ingrowth and nutrient flow) as well as mechanical properties of bone scaffolds. The ability of confocal laser scanning microscopy (CLSM) to image three‐dimensional (3‐D) structures with large vertical depths (∼2 mm) and fine vertical resolution (∼1 μm) is utilized in this article to characterize the 3‐D microstructures of hydroxyapatite (HA) bone scaffold specimens with porosity ranging from roughly 60–70 vol%. Various CLSM techniques are applied to image and interpret the HA pore structure, including Z‐series stacking, topographic profiling, and Phi‐Z scanning and contour mapping.
Decreasing xanthine oxidase‐mediated oxidative stress prevents useful cellular adaptations to exercise in rats
Authors: Gomez‐Cabrera Mari‐Carmen , Borrás Consuelo , Pallardó Federico V. , Sastre Juan , Ji Li Li , Viña Jose
Published: June 2005

Reactive oxygen or nitrogen species (RONS) are produced during exercise due, at least in part, to the activation of xanthine oxidase. When exercise is exhaustive they cause tissue damage; however, they may also act as signals inducing specific cellular adaptations to exercise. We have tested this hypothesis by studying the effects of allopurinol‐induced inhibition of RONS production on cell signalling pathways in rats submitted to exhaustive exercise. Exercise caused an activation of mitogen‐activated protein kinases (MAPKs: p38, ERK 1 and ERK 2), which in turn activated nuclear factor κB (NF‐κB) in rat gastrocnemius muscle. This up‐regulated the expression of important enzymes associated with cell defence (superoxide dismutase) and adaptation to exercise (eNOS and iNOS). All these changes were abolished when RONS production was prevented by allopurinol. Thus we report, for the first time, evidence that decreasing RONS formation prevents activation of important signalling pathways, predominantly the MAPK–NF‐κB pathway; consequently the practice of taking antioxidants before exercise may have to be re‐evaluated.
The role of NHERF‐1 in the regulation of renal proximal tubule sodium–hydrogen exchanger 3 and sodium‐dependent phosphate cotransporter 2a
Authors: Weinman Edward J. , Cunningham Rochelle , Wade James B. , Shenolikar Shirish
Published: June 2005

Adaptor proteins containing PDZ interactive domains have been recently identified to regulate the trafficking and activity of ion transporters and channels in epithelial tissue. In the renal proximal tubule, three PDZ adaptor proteins, namely NHERF‐1, NHERF‐2 and PDZK1, are expressed in the apical membrane, heterodimerize with one another, and, at least in vitro, are capable of binding to NHE3 and Npt2a, two major regulated renal proximal tubule apical membrane transporters. Studies using NHERF‐1 null mice have begun to provide insights into the organization of these adaptor proteins and their specific interactions with NHE3 and Npt2a. Experiments using brush border membranes and cultured renal proximal tubule cells indicate a specific requirement for NHERF‐1 for cAMP‐mediated phosphorylation and inhibition of NHE3. NHERF‐1 null mice demonstrate increased urinary excretion of phosphate associated with mistargeting of Npt2a to the apical membrane of renal proximal tubule cells. NHERF‐1 null animals challenged with a low phosphate diet and proximal tubule cells from these animals cultured in a low phosphate media fail to adapt as well as wild‐type mice. These studies indicate a unique requirement for NHERF‐1 in cAMP regulation of NHE3 and in the trafficking of Npt2a.
Rat subicular networks gate hippocampal output activity in an in vitro model of limbic seizures
Authors: Benini Ruba , Avoli Massimo
Published: June 2005

Evidence obtained from human epileptic tissue maintained in vitro indicates that the subiculum may play a crucial role in initiating epileptiform discharges in patients with mesial temporal lobe epilepsy. Hence, we used rat hippocampus–entorhinal cortex (EC) slices to identify the role of subiculum in epileptiform synchronization during bath application of 4‐aminopyridine (4AP, 50 μm). In these slices, fast CA3‐driven interictal‐like events were restricted to the hippocampal CA3/CA1 areas and failed to propagate to the EC where slow interictal‐like and ictal‐like epileptiform discharges were recorded. However, antagonizing GABAA receptors with picrotoxin (50 μm) made CA3‐driven interictal activity spread to EC. Sequential field potential analysis along the CA3–CA1–subiculum axis revealed that the amplitude of CA3‐driven interictal discharges recorded in the presence of 4AP only diminished within the subiculum. Furthermore, CA1 electrical stimulation under control conditions elicited little or no subicular activation and never any response in EC; in contrast, robust subicular discharges that spread to EC could be evoked after picrotoxin. Intracellular recordings indicated that potentiation by picrotoxin was associated with blockade of hyperpolarizing IPSPs in subicular cells. Finally, when surgically isolated from adjacent structures, the subiculum generated low‐amplitude synchronous discharges that corresponded to an intracellular hyperpolarization–depolarization sequence, were resistant to glutamatergic antagonists, and represented the activity of synchronized interneuronal networks. Bath application of picrotoxin abolished these 4AP‐induced events and in their place robust network bursting occurred. In conclusion, our study demonstrates that the subiculum plays a powerful gating role on hippocampal output activity. This function depends on GABAA receptor‐mediated inhibition and controls hippocampal–parahippocampal interactions that are known to modulate limbic seizures.
Biophysical and pharmacological properties of the voltage‐gated potassium current of human pancreatic β‐cells
Authors: Herrington James , Sanchez Manuel , Wunderler Denize , Yan Lizhen , Bugianesi Randal M. , Dick Ivy E. , Clark Sam A. , Brochu Richard M. , Priest Birgit T. , Kohler Martin G. , McManus Owen B.
Published: June 2005

Voltage‐gated potassium (Kv) currents of human pancreatic islet cells were studied by whole‐cell patch clamp recording. On average, 75% of the cells tested were identified as β‐cells by single cell, post‐recording RT‐PCR for insulin mRNA. In most cells, the dominant Kv current was a delayed rectifier. The delayed rectifier activated at potentials above −20 mV and had a V½ for activation of −5.3 mV. Onset of inactivation was slow for a major component (τ= 3.2 s at +20 mV) observed in all cells; a smaller component (τ= 0.30 s) with an amplitude of ∼25% was seen in some cells. Recovery from inactivation had a τ of 2.5 s at −80 mV and steady‐state inactivation had a V½ of −39 mV. In 12% of cells (21/182) a low‐threshold, transient Kv current (A‐current) was present. The A‐current activated at membrane potentials above −40 mV, inactivated with a time constant of 18.5 ms at −20 mV, and had a V½ for steady‐state inactivation of −52 mV. TEA inhibited total Kv current with an IC50= 0.54 mm and PAC, a disubstituted cyclohexyl Kv channel inhibitor, inhibited with an IC50= 0.57 μm. The total Kv current was insensitive to margatoxin (100 nm), agitoxin‐2 (50 nm), kaliotoxin (50 nm) and ShK (50 nm). Hanatoxin (100 nm) inhibited total Kv current by 65% at +20 mV. Taken together, these data provide evidence of at least two distinct types of Kv channels in human pancreatic β‐cells and suggest that more than one type of Kv channel may be involved in the regulation of glucose‐dependent insulin secretion.
Short periods of prenatal stress affect growth, behaviour and hypothalamo–pituitary–adrenal axis activity in male guinea pig offspring
Authors: Kapoor Amita , Matthews Stephen G.
Published: June 2005

Prenatal stress can have profound long‐term influences on physiological function throughout the course of life. We hypothesized that focused periods of moderate prenatal stress at discrete time points in late gestation have differential effects on hypothalamo–pituitary–adrenal (HPA) axis function in adult guinea pig offspring, and that changes in HPA axis function will be associated with modification of anxiety‐related behaviour. Pregnant guinea pigs were exposed to a strobe light for 2 h on gestational days (GD) 50, 51, 52 (PS50) or 60, 61, 62 (PS60) (gestation length ∼70 days). A control group was left undisturbed throughout pregnancy. Behaviour was assessed in male offspring on postnatal day (PND)25 and PND70 by measurement of ambulatory activity and thigmotaxis (wall‐seeking behaviour) in a novel open field environment. Subsequent to behavioural testing, male offspring were cannulated (PND75) to evaluate basal and activated HPA axis function. Body weight was significantly decreased in adult PS50 and PS60 offspring and this effect was apparent soon after weaning. The brain‐to‐body‐weight ratio was significantly increased in adult PS50 males. Basal plasma cortisol levels were elevated in PS50 male offspring throughout the 24 h sampling period compared with controls. In response to an ACTH challenge and to exposure to an acute stressor, PS60 male offspring exhibited elevated plasma cortisol responses. Plasma testosterone concentrations were strikingly decreased in PS50 offspring. Thigmotaxis in the novel environment was increased in PS50 male offspring at PND25 and PND70, suggesting increased anxiety in these animals. In conclusion, prenatal stress during critical windows of neuroendocrine development programs growth, HPA axis function, and stress‐related behaviour in adult male guinea pig offspring. Further, the nature of the effect is dependant on the timing of the maternal stress during pregnancy.