bluuebunny/crossref_metadata_2025 huggingface dataset:
Preparation and Immunolabeling of Caenorhabditis elegans: Figure 1
Authors: Crittenden Sarah , Kimble Judith
Published: May 2009

INTRODUCTION

The use of antibodies to visualize the distribution and subcellular localization of gene products powerfully complements genetic and molecular analysis of gene function in Caenorhabditis elegans. The challenge to immunolabeling C. elegans is finding the fixation and permeabilization methods that effectively make antigens accessible without destroying the tissue morphology or the antigen. Embryos are surrounded by a chitinous eggshell and larvae and adults are surrounded by a collagenous cuticle, each of which must be permeabilized to allow penetration of antibodies. In addition, antigens and antibodies are sensitive to different fixing and permeabilizing conditions. This protocol describes two methods for tissue fixation. The whole-mount freeze-cracking method is a good starting point as it is easy and works well with most antibodies and with embryos, larvae, and adults. In the tissue extrusion method, gonads and intestines, which are extruded from the carcass, are well fixed and permeabilized. Tissues remaining in the carcass are not usually stained well. The protocol concludes with an antibody incubation procedure in which fixed worms are incubated overnight with primary antibody, subsequently exposed to secondary antibody, and mounted for viewing.
Construction of Adenovirus Vectors with RGD-Modified Fiber for Transductional Targeting
Authors: Kawakami Yosuke , Curiel David T.
Published: May 2009

INTRODUCTION

Therapeutic gene transfer strategies aim to maximize gene transfer and expression in target cells. However, gene delivery systems often fail to preferentially transduce target cells in mixed cell populations. In addition, limitations in vector specificity can lead to transduction of non-target cells, resulting in untoward toxicity, even with compartmental dosing. Thus, vector optimization is critical for the development of efficient genetic experiments. The life cycle and biology of adenovirus (Ad) infection and transgene expression in cells have been thoroughly characterized. Ad infection is initiated by recognition of the native Ad5 receptor, coxsackievirus-adenovirus receptor (CAR), on target cells by the carboxy-terminal portion (i.e., knob) of the fiber protein. The development of genetically modified Ad vectors with transductional specificity for a single cell type requires “retargeting”: the ablation of endogenous tropism and the introduction of novel tropism determinants for target cells. This protocol describes the construction of chimeric fibers. Homologous recombination between the pAdEasy-1 and the fiber shuttle plasmid (pFiber-dE3-RGD) is used to generate a rescue plasmid with a mutant fiber that inserts Arg-Gly-Asp (RGD) at the carboxyl terminus of the fiber knob region. The resultant fiber-modified rescue plasmid (pAdEasy-RGD) is isogenic to AdEasy-1 (except the fiber region) and can therefore be used for another round of homologous recombination with pShuttle-promoter-Luc to make a final Ad vector construct that expresses the luciferase gene and contains an RGD-modified fiber knob. This construct can then be tested for targeting efficacy against cells expressing αv integrin cell surface receptors.
Systematic Monitoring of Protein Complex Composition and Abundance by Blue-Native PAGE
Authors: Eubel Holger , Millar A. Harvey
Published: May 2009

INTRODUCTION

Many polypeptides do not perform their functions as single autonomous units in vivo. Instead, multiple polypeptides associate to form higher molecular mass structures. Blue-native polyacrylamide gel electrophoresis (BN-PAGE) allows a range of the major protein complexes involved in such protein-protein interactions to be visualized simultaneously and in a single experiment. When combined with a second dimension of sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), the BN/SDS-PAGE procedure can resolve the complexes according to their molecular weight, as well as the subunits within each complex, according to the molecular weights of the subunits. Similarly, used in conjunction with differential in-gel electrophoresis (DIGE), it can accurately quantify changes in protein complex abundance or subunit composition between different samples, or between different complexes within the same sample. The following basic protocol describes sample preparation and gel casting for the first (BN-PAGE) and second (SDS-PAGE) dimensions. Variants are presented with and without DIGE labeling, along with the additional steps required for the fluorescence DIGE technique.
The Cytokinin Type-B Response Regulator PtRR13 Is a Negative Regulator of Adventitious Root Development inPopulus
Authors: Ramírez-Carvajal Gustavo A. , Morse Alison M. , Dervinis Christopher , Davis John M.
Published: April 2009

Adventitious root formation at the base of plant cuttings is an innate de novo organogenesis process that allows massive vegetative propagation of many economically and ecologically important species. The early molecular events following shoot excision are not well understood. Using whole-genome microarrays, we detected significant transcriptome remodeling during 48 h following shoot removal in Populus tremula × Populus alba softwood cuttings in the absence of exogenous auxin, with 27% and 36% of the gene models showing differential abundance between 0 and 6 h and between 6 and 24 h, respectively. During these two time intervals, gene networks involved in protein turnover, protein phosphorylation, molecular transport, and translation were among the most significantly regulated. Transgenic lines expressing a constitutively active form of the Populus type-B cytokinin response regulator PtRR13 (ΔDDKPtRR13) have a delayed rooting phenotype and cause misregulation of CONTINUOUS VASCULAR RING1, a negative regulator of vascularization; PLEIOTROPIC DRUG RESISTANCE TRANSPORTER9, an auxin efflux transporter; and two APETALA2/ETHYLENE RESPONSE FACTOR genes with sequence similarity to TINY. Inappropriate cytokinin action via ΔDDKPtRR13 expression appeared to disrupt adventitious root development 24 h after shoot excision, when root founder cells are hypothesized to be sensitive to the negative effects of cytokinin. Our results are consistent with PtRR13 acting downstream of cytokinin to repress adventitious root formation in intact plants, and that reduced cytokinin signaling after shoot excision enables coordinated expression of ethylene, auxin, and vascularization pathways leading to adventitious root development.
BIN2 Functions Redundantly with Other Arabidopsis GSK3-Like Kinases to Regulate Brassinosteroid Signaling
Authors: Yan Zhenyan , Zhao Jun , Peng Peng , Chihara Ray K. , Li Jianming
Published: April 2009

GLYCOGEN SYNTHASE KINASE3 (GSK3) is a highly conserved serine/threonine kinase involved in a variety of developmental signaling processes. The Arabidopsis (Arabidopsis thaliana) genome encodes 10 GSK3-like kinases that are clustered into four groups. Forward genetic screens have so far uncovered eight mutants, all of which carry gain-of-function mutations in BRASSINOSTEROID-INSENSITIVE2 (BIN2), one of the three members in group II. Genetic and biochemical studies have implicated a negative regulatory role for BIN2 in brassinosteroid (BR) signaling. Here, we report the identification of eight ethyl methanesulfonate-mutagenized loss-of-function bin2 alleles and one T-DNA insertional mutation each for BIN2 and its two closest homologs, BIN2-Like1 and BIN2-Like2. Our genetic, biochemical, and physiological assays revealed that despite functional redundancy, BIN2 plays a dominant role among the three group II members in regulating BR signaling. Surprisingly, the bin2bil1bil2 triple T-DNA insertional mutant still responds to BR and accumulates a more phosphorylated form of a BIN2 substrate than the wild-type plant. Using the specific GSK3 inhibitor lithium chloride, we have provided strong circumstantial evidence for the involvement of other Arabidopsis GSK3-like kinases in BR signaling. Interestingly, lithium chloride treatment was able to suppress the gain-of-function bin2-1 mutation but had a much weaker effect on a strong BR receptor mutant, suggesting the presence of a BIN2-independent regulatory step downstream of BR receptor activation.
The Wound-, Pathogen-, and Ultraviolet B-ResponsiveMYB134Gene Encodes an R2R3 MYB Transcription Factor That Regulates Proanthocyanidin Synthesis in Poplar
Authors: Mellway Robin D. , Tran Lan T. , Prouse Michael B. , Campbell Malcolm M. , Constabel C. Peter
Published: April 2009

In poplar (Populus spp.), the major defense phenolics produced in leaves are the flavonoid-derived proanthocyanidins (PAs) and the salicin-based phenolic glycosides. Transcriptional activation of PA biosynthetic genes leading to PA accumulation in leaves occurs following herbivore damage and mechanical wounding as well as infection by the fungal biotroph Melampsora medusae. In this study, we have identified a poplar R2R3 MYB transcription factor gene, MYB134, that exhibits close sequence similarity to the Arabidopsis (Arabidopsis thaliana) PA regulator TRANSPARENT TESTA2 and that is coinduced with PA biosynthetic genes following mechanical wounding, M. medusae infection, and exposure to elevated ultraviolet B light. Overexpression of MYB134 in poplar resulted in transcriptional activation of the full PA biosynthetic pathway and a significant plant-wide increase in PA levels, and electrophoretic mobility shift assays showed that recombinant MYB134 protein is able to bind to promoter regions of PA pathway genes. MYB134-overexpressing plants exhibited a concomitant reduction in phenolic glycoside concentrations and other minor alterations to levels of small phenylpropanoid metabolites. Our data provide insight into the regulatory mechanisms controlling stress-induced PA metabolism in poplar, and the identification of a regulator of stress-responsive PA biosynthesis constitutes a valuable tool for manipulating PA metabolism in poplar and investigating the biological functions of PAs in resistance to biotic and abiotic stresses.
Energy-dispersive diffraction studies of inert anodes
Authors: Scarlett Nicola V. Y. , Madsen Ian C. , Evans John S. O. , Coelho Alan A. , McGregor Katherine , Rowles Matthew , Lanyon Marshall R. , Urban Andrew J.
Published: May 2009

A tomographic study of electrochemical cells to observe scales formed on inert anodes has been conducted using energy-dispersive synchrotron X-ray diffraction. This study is preparatory to an investigation that will observe this formationin situduring the cells' operation. The purpose of the current work was to determine whether this technique would be appropriate for such a study in terms of its sensitivity and whether the results could be quantified satisfactorily. A method has been developed for the quantitative phase analysis of energy-dispersive data using crystal-structure-based Rietveld refinement. This has been tested with standard materials and found to be comparable in accuracy to results obtained from traditional angular-dispersive diffraction. The lower limits of detection of the method have not been established quantitatively but qualitative differences can be seen between cells that have been cycled at different times. These differences indicate a linear relationship between scale formation and electrolysis time.
Structures of the ZrZn22 family: suprapolyhedral nanoclusters, methods of self-assembly and superstructural ordering
Authors: Ilyushin G. D. , Blatov V. A.
Published: May 2009

A combinatorial topological analysis is carried out by means of the program package TOPOS4.0 [Blatov (2006), IUCr Comput. Commun. Newsl.
7, 4–38] and the matrix self-assembly is modeled for crystal structures of the ZrZn22 family (space group Fd\bar 3m, Pearson code cF184), including the compounds with superstructural ordering. A number of strict rules are proposed to model the crystal structures of intermetallics as a network of cluster precursors. According to these rules the self-assembly of the ZrZn22-like structures was considered within the hierarchical scheme: primary polyhedral cluster → zero-dimensional nanocluster precursor → one-dimensional primary chain → two-dimensional microlayer → three-dimensional microframework (three-dimensional supraprecursor). The suprapolyhedral cluster precursor AB
2
X
37 of diameter ∼ 12 Å and volume ∼ 350 Å3 consists of three polyhedra (one AX
16 of the \bar 43m point symmetry and two regular icosahedra BX
12 of the \bar 3m point symmetry); the packing of the clusters determines the translations in the resulting crystal structure. A novel topological type of the two-dimensional crystal-forming 4,4-coordinated binodal net AB
2, with the Schläfli symbols 3636 and 3366 for nodes A and B, is discovered. It is shown that the ZrZn22 superstructures are formed by substituting some atoms in the cluster precursors. Computer analysis of the CRYSTMET and ICSD databases shows that the cluster AB
2
X
37 occurs in 111 intermetallics belonging to 28 structure types.
Scout robot with wheeling‐hopping combination locomotion
Authors: Zhao Jie , Liu Gangfeng , Yan Jihong , Zang Xizhe
Published: May 2009

Purpose

The purpose of this paper is to present a new scout robot that tries to combine the hopping movement and the wheeling movement to greatly enlarge the scope of robot's activities.

Design/methodology/approach

A five‐shank hopping mechanism was employed to build the wheeling‐hopping combination scout robot. The non‐linear character of the five‐shank hopping mechanism was analyzed and then used in the proposed non‐linear spring‐mass model for the robot.

Findings

The rules of robot's movement were deduced, influencing factors of the jumping height were analyzed and the countermeasure was adopted. Simulations and an experiment of the robot's movement showed that the robot has strong locomotivity and survival ability.

Originality/value

A five‐shank hopping mechanism is proposed, analyzed and combined with wheeling movement to enhance the locomotivity and survival ability of scout robot.
Inspection of welding pool height from shading in pulsed GTAW with wire filler
Authors: Wang J.F. , Wang W.Y. , Chen S.B.
Published: May 2009

Purpose

The control of weld penetration in gas tungsten‐arc welding (GTAW) is required for fully automated systems to overcome variations in the welding process. The surface depression of weld pool, or weld pool height, has a close relationship with width of the backside bead in the full‐penetrated weld. The purpose of this paper is to inspect the pool height.

Design/methodology/approach

A fast linear approach, based on shape from shading (SFS) algorithm, was employed to reconstruct a 3D shape of welding pool from a 2D image, which was obtained from a real welding pool. Then the pool height can be extracted. Furthermore, three methods were introduced to improve the above algorithm.

Findings

The reconstructed pool height was in good agreement with the real height of weld pool from experiments.

Research limitations/implications

The algorithm requires a uniform reflection on the pool surface.

Practical implications

This method is applicable to inspect weld pool height in GTAW. It is a basis for future work on control of weld penetration.

Originality/value

A faster SFS algorithm has been introduced to extract weld pool height in real‐time. Moreover, the algorithm was improved to fit for extracting the surface shape of weld pool.
Team‐based reward allocation structures and the helping behaviors of outcome‐interdependent team members
Authors: Bamberger Peter A. , Levi Racheli
Published: May 2009

Purpose

The purpose of this paper is to examine the effects of two key team‐based pay characteristics – namely reward allocation procedures (i.e. reward based on norms of equity, equality or some combination of the two) and incentive intensity – on both the amount and type of help given to one another among members of outcome‐interdependent teams.

Design/methodology/approach

A total of 180 undergraduate students participate in a laboratory simulation with a 2 × 3 experimental design. Servicing virtual “clients,” participants receive pre‐scripted requests for assistance from anonymous teammates. ANOVA and hierarchical regression analyses are used to test the hypotheses.

Findings

Relative to equity‐oriented group‐based pay structures, equality‐oriented pay structures are found to be associated with both significantly more help giving in general and more of the type of help likely to enhance group‐level competencies (i.e. autonomous help). Incentive intensity strengthens the effects of reward allocation on the amount (but not the type) of help giving.

Research limitations/implications

While the short time frame of the simulation poses a significant threat to external validity, the findings suggest that team‐based compensation practices may provide organizational leaders with an important tool by which to shape critical, helping‐related team processes, with potentially important implications for both team learning and performance.

Practical implications

Managers interested in promoting capacity‐building and helping among team members should avoid allocating team rewards strictly on the basis of the individual contribution.

Originality/value

This paper provides the first empirical findings regarding how alternative modes of team‐based reward distribution may influence key group processes among members of outcome interdependent teams.
Effects of country variables on young generation's attitude towards American products: a multi‐attribute perspective
Authors: Chen Hsiu‐Li
Published: May 2009

Purpose

The purpose of this paper is to examine the country‐of‐origin (COO) effect and consumer patriotism on young generation's attitude toward American products with multi‐attributes: across different cultures and different product categories.

Design/methodology/approach

A 2 × 2 research design (Taiwan/Indonesia)×(high‐tech products and non‐high‐tech products) and LISREL technique are employed to examine the effects.

Findings

The results of the study find that both Taiwanese and Indonesian cases show that country‐of‐origin (COO) has a significantly positive effect on the attitude toward American‐made products. Moreover, Indonesian respondents who demonstrate higher patriotism show negative attitudes toward American products. The effects of COO on purchase intentions are product‐specific and consumer consuming‐specific. Stronger product attitude exists for Taiwan's sample than for Indonesia's, primarily due to higher cultural identification with America.

Originality/value

The “Made in USA” slogan could be used to promote American products to people from younger generations. COO may have a different effect on consumer purchase intention between high‐tech and non‐high‐tech products and between different consuming countries. Marketers using the “Made in USA” slogan as a promotion tool should show special concern for the product category as well as for cultural identification factors.
Analysing complex policy change in Hong Kong: what role for critical discourse analysis?
Authors: Cheng Annie Y.N.
Published: May 2009

Purpose

The Hong Kong government recently reformed pre‐primary education with the introduction of a voucher scheme. At the time this policy caused considerable opposition from across Hong Kong Society. This paper seeks to use Fairclough's model of critical discourse analysis to explore a key policy text and seeks to assess to what extent such an analytical framework can help develop a better understanding of the processes of policy development.

Design/methodology/approach

The paper uses critical discourse analysis methods and applies these to a specific policy text relating to the reform of pre‐primary education.

Findings

The paper suggests that critical discourse analysis can provide a useful analytical tool for analysing the complexities of the policy development process. In particular it surfaces key tensions within policy, such as those between commitments to equity and efficiency, that may have a significant impact on the outcomes of subsequent policy implementation.

Research limitations/implications

The paper is based on a small scale study. Its findings are tentative. There is some merit in the approach adopted, but the use of this approach in relation to policy development requires further work to develop it.

Originality/value

The paper adds to the understanding of how a range of methods of data analysis can help develop a more sophisticated understanding of the nuances of policy development.
Assessment of quality of midwifery care in labour and delivery wards of selected Kordestan Medical Science University hospitals
Authors: Simbar Masoumeh , Ghafari Farideh , Tork Zahrani Shahnaz , Alavi Majd Hamid
Published: May 2009

Purpose

Quality improvement of reproductive health care has been announced as one of five global strategies to accelerate progress toward reproductive health goals. The World Health Organization emphasises the evaluation of structure, procedure and outcome of health services to improve quality of care. This study aims to assess the quality of provided care in labour and delivery units in two selected Kordestan Medical Science University hospitals.

Design/methodology/approach

A descriptive study methodology was utilised to assess the quality of care provided to 96 women with normal pregnancies. Two checklists were used to observe procedures of care and structure together with a questionnaire utilised to assess satisfaction ratings of patients. Data were analysed by SPSS 11.5.

Findings

Midwifery care was provided in different stages of labour, with the following mean percentages of compatibility with desirable situation: first stage of labour (71.4 per cent), second stage of labour (63.03 per cent), third stage of labour (80.63 per cent) and first 2 hours after labour (70.50 per cent). The lowest scores were related to the domains of “emotional support”, “hand wash” and “assessment of vital signs”.

Originality/value

The paper develops instructions for care provision or promotion of partograph use for continuous monitoring and evaluation of quality of care by managers. Increasing midwifery personnel and providing facilities for accompanied people to improve quality of emotional care can lead to quality improvement, and finally the women's health and satisfaction.
Nascent entrepreneurial activity within female ethnic minority groups
Authors: Kwong Caleb C.Y. , Thompson Piers , Jones‐Evans Dylan , Brooksbank David
Published: May 2009

Purpose

The purpose of this paper is to compare the entrepreneurial activity, attitudes and social connections of four groups of ethnic minority females in the UK, with the aim of examining the extent of gender and ethnic background effects on nascent start‐up activities and the attitudes of women belonging to these ethnic minority groups.

Design/methodology/approach

A two‐stage approach is adopted to examine the situations of four main female ethnic minority groups using data from the Global Entrepreneurship Monitor (GEM) adult population survey for the UK. The first stage adopts a binary logistic approach to determine the importance of social networks, opportunity perception and risk aversion to the probability of being involved in nascent entrepreneurial activities. The second stage of analysis examines the differences in these perceptual variables to determine the extent to which different female ethnic minority groups are embedded in different social environments when attempting to start a business.

Findings

There are considerable differences amongst different ethnic groups in the level of entrepreneurial activity by women, their attitudes towards entrepreneurship, and the social capital available to them when starting a business.

Practical implications

The findings suggest that policy makers should take the differences by ethnic groupings into account when developing bespoke development policies designed to alleviate the barriers faced by women.

Originality/value

The paper is one of the first comparative studies focusing on women from different ethnic backgrounds. Rather than assuming homogeneity, or examining specific groups in isolation it allows the different conditions faced by prospective entrepreneurs from each group to be examined.
New tensions and challenges in integrated communications
Authors: Thøger Christensen Lars , Fuat Fırat A. , Cornelissen Joep
Published: May 2009

Purpose

The purpose of this paper is to demonstrate how tensions and challenges associated with the implementation of integrated communications in practice have intensified in recent years under the impact of two conflicting trends: new social and organizational “drivers” towards integration; and the appearance of savvy and sophisticated audiences.

Design/methodology/approach

Taking the point of departure in now classical discussions of structural “barriers” to integration, today more fundamental difficulties limit the implementation of integrated communications – difficulties rooted in epistemological issues of organization and communication are argued.

Findings

Integrated communications present a paradox to contemporary communication management. On the one hand, integration seems to be the most logical and sensible way of managing communications in a complex world of multiple and critical audiences. On the other hand, its prescriptions are essentially at odds with what is known today about organization and human communication. While representing a significant communication ideal, integration in communications is simultaneously beyond reach – at least if the full process of communication is taken into proper consideration.

Originality/value

The paper usefully makes the claim that it needs to adapt the ideals of what integrated communications is to what integration can possibly do to an organization and its communications.
Silicon Schottky barrier photodiodes with a thin AlN nucleation layer
Authors: Chuah L.S. , Hassan Z. , Abu Hassan H. , Chin C.W. , Thahab S.M. , Teoh S.C.
Published: May 2009

Purpose

The purpose of this paper is to present the characteristics of novel silicon Schottky barrier (SB) photodiodes (PDs) with aluminium nitride (AlN) (100 nm) nucleation layer.

Design/methodology/approach

Comparison was made with conventional silicon SB PDs.

Findings

It was found that smaller dark current could be achieved with AlN nucleation layer. It was also found that effective SB height increased from 0.65 to 0.71 eV with the insertion of the AlN layer. The dark leakage current for the Schottky PDs with the AlN layer was shown to be about two orders of magnitude smaller than that for the conventional silicon SB PDs.

Research limitations/implications

It is possible that the detrimental effect of interface states situated near the metal semiconductor interface was less pronounced for the sample owing to the insertion of the AlN nucleation layer.

Originality/value

There is believed to be no other report on silicon SB PDs capped with an AlN layer in the literature. This paper describes the fabricated silicon SB PDs and reports on the electrical characteristics of the devices with an AlN nucleation layer grown at low temperature.
Ultrasound vs nerve stimulation multiple injection technique for posterior popliteal sciatic nerve block
Authors: Danelli G. , Fanelli A. , Ghisi D. , Moschini E. , Rossi M. , Ortu A. , Baciarello M. , Fanelli G.
Published: May 2009

In this prospective, randomised, observer‐blinded study we evaluated whether ultrasound guidance can shorten the onset time of popliteal sciatic nerve block as compared to nerve stimulation with a multiple injection technique. Forty‐four ASA I–III patients undergoing posterior popliteal sciatic nerve block with 20 ml of 0.75% ropivacaine were randomly allocated to nerve stimulation or ultrasound guided nerve block. A blinded observer recorded onset of sensory and motor blocks, success rates, the need for fentanyl intra‐operatively, the requirement for general anaesthesia, procedure‐related pain, patient satisfaction and side‐effects. Onset times for sensory and motor blocks were comparable. The success rate was 100% for ultrasound guided vs 82% for nerve stimulation (p = 0.116). Ultrasound guidance reduced needle redirections (p = 0.01), were associated with less procedural pain (p = 0.002) and required less time to perform (p = 0.002). Ultrasound guidance reduced the time needed for block performance and procedural pain.
Association of insulin resistance with anti‐Mullerian hormone levels in women without polycystic ovary syndrome (PCOS)
Authors: Park Hyun T. , Cho Geum J. , Ahn Ki H. , Shin Jung H. , Kim Young T. , Hur Jun Y. , Kim Sun H. , Lee Kyu W. , Kim Tak
Published: April 2009

Objective  To explore the relationship of insulin resistance (IR) and adipokines (leptin, adiponectin, RBP4) to anti‐Mullerian hormone (AMH) levels in women without polycystic ovary syndrome (PCOS).

Design/patients/measurements  We recruited 120 healthy, reproductive age women without PCOS. An overnight fasting blood draw, anthropometric measurements, analyses of serum levels of AMH, adipokines (leptin, adiponectin and RBP4) and total testosterone, a homeostasis model assessment for insulin resistance (HOMA‐IR) and a transvaginal ultrasound scan were performed between the third and fifth day of their spontaneous menstrual cycles.

Results  Higher HOMA‐IR levels were associated with lower levels of AMH. After adjustment for age, serum AMH levels negatively correlated with insulin, fasting glucose, HOMA‐IR and RBP4. However, a positive correlation was identified between serum AMH and adiponectin. A final multiple stepwise linear regression demonstrated that HOMA‐IR was independently associated with AMH.

Conclusion  An independent relationship exists between HOMA‐IR and AMH in women without PCOS, possibly due to the effect of abnormal insulin action on AMH secretion by granulosa cells.
Association of PPARγ allelic variation, osteoprotegerin and abdominal aortic aneurysm
Authors: Moran Corey S. , Clancy Paula , Biros Erik , Blanco‐Martin Bernardo , McCaskie Pamela , Palmer Lyle J. , Coomans Danny , Norman Paul E. , Golledge Jonathan
Published: April 2009

Objective  We have previously demonstrated high concentrations of the glycoprotein osteoprotegerin (OPG) in biopsies of abdominal aortic aneurysm (AAA), and demonstrated that ligation of the nuclear receptor peroxisome proliferator‐activated receptor gamma (PPARγ) downregulates OPG in vitro and within a mouse model. The aims of this study were to assess the associations between circulating concentrations of OPG, polymorphisms of the gene encoding PPARγ (PPARG), AAA presence and growth.

Design, patients and measurements  Two genetic polymorphisms in PPARG were assessed in 4227 men, 699 of whom had an AAA. For 631 men, who had AAAs, maximum aortic diameter was monitored by yearly ultrasound for a median of 5 years. Plasma OPG was measured in 838 men, 318 of whom had an AAA.

Results  Plasma concentrations of OPG were independently associated with AAA (adjusted odds ratio 1.38, 95% CI 1.10–1.72). The PPARG c.1347C > T polymorphism was associated with plasma concentrations of OPG (beta 0.12, P < 0.01). The PPARG c.34G > C polymorphism was weakly associated with AAA (adjusted odds ratio 1.28, 95% CI 1.01–1.61). PPARG c.1347C > T was associated with increased AAA growth (recessive model, P = 0.03).

Conclusions  Circulating concentrations of osteoprotegerin are associated with abdominal aortic aneurysm and with one peroxisome proliferator‐activated receptor gamma gene polymorphism. Peroxisome proliferator‐activated receptor gamma gene polymorphisms are weakly associated with abdominal aortic aneurysm presence and growth. Confirmation of these findings is required in other cohorts.
Stability in temperate reef communities over a decadal time scale despite concurrent ocean warming
Authors: STUART‐SMITH RICK D. , BARRETT NEVILLE S. , STEVENSON DAVID G. , EDGAR GRAHAM J.
Published: April 2009

Despite increasing scientific and public concerns on the potential impacts of global ocean warming on marine biodiversity, very few empirical data on community‐level responses to rising water temperatures are available other than for coral reefs. This study describes changes in temperate subtidal reef communities over decadal and regional scales in a location that has undergone considerable warming in recent decades and is forecast to be a ‘hotspot’ for future warming.

Plant and animal communities at 136 rocky reef sites around Tasmania (south‐east Australia) were censused between 1992 and 1995, and again in 2006 and 2007. Despite evidence of major ecological changes before the period of study, reef communities appeared to remain relatively stable over the past decade. Multivariate analyses and univariate metrics of biotic communities revealed few changes with time, although some species‐level responses could be interpreted as symptomatic of ocean warming. These included fishes detected in Tasmania only in recent surveys and several species with warmer water affinities that appeared to extend their distributions further south. The most statistically significant changes observed in species abundances, however, were not related to their biogeographical affinities. The majority of species with changing abundance possessed lower to mid‐range abundances rather than being common, raising questions for biodiversity monitoring and management. We suggest that our study encompassed a relatively stable period following more abrupt change, and that community responses to ocean warming may follow nonlinear, step‐like trajectories.
Do helminths cause epilepsy?
Authors: WAGNER R. G. , NEWTON C. R.
Published: April 2009

Both helminthiases and epilepsy occur globally, and are particularly prevalent in developing regions of the world. Studies have suggested an association between epilepsy and helminth infection, but a causal relationship is not established in many helminths, except perhaps with neurocysticercosis. We review the available literature on the global burden of helminths, and the epidemiological evidence linking helminths to epilepsy. We discuss possible routes that helminths affect the central nervous system (CNS) of humans and the immunological response to helminth infection in the CNS, looking at possible mechanisms of epileptogenesis. Finally, we discuss the current gaps in knowledge about the interaction between helminths and epilepsy.
Variants of the IL23R Gene are Associated with Ankylosing Spondylitis but not with Sjögren Syndrome in Hungarian Population Samples
Authors: Sáfrány E. , Pazár B. , Csöngei V. , Járomi L. , Polgár N. , Sipeky C. , Horváth I. F. , Zeher M. , Poór G. , Melegh B.
Published: April 2009

Recently, associations were found between several autoimmune diseases and functional variants of interleukin‐23 receptor (IL23R) gene; here, we studied the possible association of nine polymorphisms of IL23R with ankylosing spondylitis (AS) and with Sjögren syndrome (SS). In our study, we genotyped groups of patients with AS (n = 206), SS (n = 156) and healthy controls (n = 235) for rs11805303, rs10889677, rs1004819, rs2201841, rs11209032, rs11209026, rs10489629, rs7517847 and rs7530511 variants using PCR‐RFLP methods. We observed significant increase in the carriage of the T allele of rs11805303 and the A allele of rs1004189 in the AS group compared with the controls. For the rs10889677 variant, the prevalence of the AA genotype and for the rs2201841, the CC genotype showed a more than two‐fold increase in the AS group compared with the controls. By contrast, the GA heterozygous genotype of rs11209026 variant showed a significant decrease in AS patients compared with controls. Haplotype analysis revealed association of four IL23R haplotypes with AS. There was no difference in the distribution of any of the examined IL23R variants between controls and SS patients. In conclusion, we confirmed the susceptibility or protective associations of IL23R polymorphisms with AS in a Hungarian population and first demonstrated the involvement of the rs11805303 intronic single nucleotide polymorphisms, which was tested so far only for other autoimmune diseases.
Cell Specific Synovial Expression of Nicotinic Alpha 7 Acetylcholine Receptor in Rheumatoid Arthritis and Psoriatic Arthritis
Authors: Westman M. , Engström M. , Catrina A. I. , Lampa J.
Published: April 2009

Neuroimmune interactions are known to influence several chronic inflammatory and rheumatic diseases, but the underlying mechanisms have been insufficiently elucidated. The cholinergic anti‐inflammatory pathway is characterized by neural regulation of systemic inflammation, mediated by the vagus nerve and specific cholinergic stimulation of the nicotinic α‐7 acetylcholine receptor (α7nAChR) on immune cells. Moreover, α7nAChR has been shown important for immune regulation also in the absence of nerves, but little is known about these mechanisms in chronic joint inflammation. The expression and localization of α7nAChR in synovial biopsies from patients with rheumatoid arthritis and psoriatic arthritis was investigated by immunohistochemistry using monoclonal antibody against α7nAChR. Surface staining of α7nAChR was observed in synovial tissue of all arthritis patients investigated and could also to a lesser extent be detected in the synovium of healthy individuals. α7nAChR positive cells were detected in mainly synovial lining cells and vessels. The α7nAChR positively stained cells were by double immunofluorescence identified as primarily macrophages and fibroblasts, with the majority of these cells expressing the receptor. These results indicate the importance of α7nAChR and cholinergic mechanisms in arthritis pathogenesis and implicate specific cholinergic modulation as a potential anti‐inflammatory therapeutic strategy in joint inflammation.
CD25 Shedding by Human Natural Occurring CD4+CD25+ Regulatory T Cells does not Inhibit the Action of IL‐2
Authors: Pedersen A. E. , Lauritsen J. P.
Published: April 2009

Regulatory T (Treg) cells are important for the maintenance of peripheral tolerance and inhibition of pathogenic T‐cell responses. Therefore, they are important for the limitation of chronic inflammation but can also be deleterious by e.g. limiting antitumour immune responses. Natural occurring Tregs are known to inhibit CD4+ T cell in a contact‐dependent manner, but at the same time, various suppressive factors are secreted. We, here, demonstrate that human naturally occurring CD4+CD25+ Tregs are able to shed large amounts of soluble CD25 upon activation. Secretion of sCD25 could add to the inhibitory effect of Tregs as such secretion in other settings has been proposed to act as a sink for local IL‐2. However, we here demonstrate that supernatant from human Tregs containing high concentration of sCD25 does not inhibit proliferation of CD4+CD25− T cells or inhibit the action of IL‐2 in an in vitro bioassay.
Vaccination with β2‐Microglobulin‐Deficient Dendritic Cells Protects Against Growth of β2‐Microglobulin‐Deficient Tumours
Authors: Dammeyer P. , Mwakigonja A. R. , Rethi B. , Chiodi F. , Wolpert E. Z.
Published: April 2009

Defects in cell surface expression of major histocompatibility complex class I antigen molecules are common in tumour cells. We have previously described the generation of adaptive immunity to tumour cells deficient in the transporter associated with antigen processing molecule. In this study, we demonstrate enhanced in vivo protection against growth of β2‐microglobulin‐deficient tumour cells in syngeneic C57Bl/6 mice, following vaccination with β2‐microglobulin‐deficient dendritic cells. In vitro analysis suggested that vaccinated mice produced CD3+ cells, which could induce apoptosis in syngeneic β2‐microglobulin‐deficient tumour and non‐malignant cells. Further investigation of target cell recognition suggested that also tumour cells lacking expression of classical major histocompatibility complex class I heavy chains and functional transporter associated with antigen processing molecules were recognized by CD3+ effector cells from vaccinated mice. Histopathological examination of organs from vaccinated mice showed no significant vaccination‐induced pathology. The present findings point to a new possible strategy to counteract the growth of major histocompatibility complex class I‐deficient tumour cells.
Increase in Human Endogenous Retrovirus HERV‐K (HML‐2) Viral Load in Active Rheumatoid Arthritis
Authors: Reynier F. , Verjat T. , Turrel F. , Imbert P. E. , Marotte H. , Mougin B. , Miossec P.
Published: April 2009

To study the viral loads of human endogenous retrovirus HERV‐K (HML‐2) type 1 and type 2 in rheumatoid arthritis (RA), we measured the viral loads of HERV‐K (HML‐2) type 1 and type 2 using nucleic acid sequence‐based amplification (NASBA) technology. We analyzed plasma samples from RA patients (n = 79) and healthy volunteers (HV, n = 46) and synovial fluid samples from RA (n = 10) and osteoarthritis (OA, n = 10) patients. HERV‐K type 1 and type 2 viruses were detected and quantified for the majority of plasma and synovial fluid samples from RA patients. HERV‐K type 1 and type 2 viral loads were significantly elevated in RA patients compared with HV in plasma (P < 0.0001) and from RA patients compared with OA patients in synovial fluid (type 1: P = 0.0007; type 2: P = 0.023). Moreover, an association was observed between the HERV‐K type 1 viral load in plasma and the disease activity in RA patients (RA patients with low activity versus high activity P = 0.0129; RA patients with intermediate activity versus high activity P = 0.037). Our findings showed that HERV‐K (HML‐2) viral load can be detected in plasma samples from RA patients, with higher levels observed for those with active disease. There was an association of HERV‐K type 1 levels with the disease activity.
Suppression of Lymphocyte Proliferation and Regulation of Dendritic Cell Phenotype by Soluble Mediators from Rat Lacrimal Epithelial Cells
Authors: De Saint Jean M. , Nakamura T. , Wang Y. , Trousdale M. D. , Schechter J. E. , Mircheff A. K.
Published: April 2009

Lacrimal epithelial cells appear to constitutively secrete autoantigens to their underling stroma. The present experiments address the hypothesis that they also secrete soluble factors that regulate immune responses. Epithelial cells, spleen cells and lymphocytes were obtained from rabbits or rats and cultured in various configurations. Monocytes from rat bone marrow were matured to dendritic cells (DC) ex vivo. Proliferation was measured by [3H]‐thymidine incorporation; surface MHC Class II and CD86 using flow cytometry; and mRNA relative abundances using real time RT‐PCR. Microporous culture inserts containing rat lacrimal cells inhibited proliferation of rabbit lymphocytes co‐cultured with autologous lacrimal cells and of rat lymphocytes co‐cultured with TNF‐α‐stimulated DC. They inhibited CD86 and MHC Class II surface expression by maturating DC and reversed surface expression of CD86 but not MHC Class II by partially matured DC. Subsequent exposure of partially matured DC to mediators from rat lacrimal cells reversed the ability to stimulate lymphocyte proliferation. TGF‐β1 and IL‐10 mRNAs increased somewhat when rat lacrimal cells were isolated but decreased markedly in rabbit lacrimal cells. Antibodies to TGF‐β prevented soluble factors from rat lacrimal cells from inhibiting proliferation of rabbit lymphocytes co‐cultured with rabbit lacrimal cells, but recombinant TGF‐β alone did not mimic the soluble factors. IL‐10 immunopositivity was detected in epithelial cells of interlobular ducts and occasional interstitial cells in rabbit lacrimal gland. Rat lacrimal epithelial cells secrete TGF‐β and other factors that synergize to suppress lymphocyte proliferation and regulate DC maturation. Interlobular duct epithelial cells in rabbit lacrimal glands may express similar functions.
Impaired Antibody Responses but Normal Proliferation of Specific CD4+ T Cells in Mice Lacking Complement Receptors 1 and 2
Authors: Carlsson F. , Getahun A. , Rutemark C. , Heyman B.
Published: April 2009

Severely impaired Ab responses are seen in animals lacking C (complement) factors C2, C3 or C4 as well as CR1/2 (C receptors 1 and 2). The molecular mechanism behind this phenomenon is not understood. One possibility is that C‐containing immune complexes are endocytosed via CR2 on B cells and presented to specific CD4+ T cells, which would then proliferate and provide efficient help to specific B cells. In vitro, B cells can endocytose immune complexes via CR1/2 and present the Ag to T cells. Whether absence of this Ag presenting function in Cr2−/− mice (mice lacking CR1/2) explains their low Ab response is unclear. To address this question, Cr2−/− and wild type mice were transferred with OVA‐specific T cells, obtained from the DO11.10 strain which has a transgenic TCR recognizing an OVA peptide. The animals were subsequently immunized with sheep red blood cells (SRBC) conjugated to OVA. Interestingly, proliferation of the OVA‐specific T cells was normal in Cr2−/− mice, although their Ab response to both SRBC and OVA was severely impaired. These observations suggest that the impaired Ab response in Cr2−/− mice cannot be explained by a lack of appropriate induction of T cell help.
Quantitative measurement of specific biomarkers for protein oxidation, nitration and glycation in Arabidopsis leaves
Authors: Bechtold Ulrike , Rabbani Naila , Mullineaux Philip M. , Thornalley Paul J.
Published: April 2009

Higher plants are continually exposed to reactive oxygen and nitrogen species during their lives. Together with glucose and reactive dicarbonyls, these can modify proteins spontaneously, leading to protein oxidation, nitration and glycation. These reactions have the potential to damage proteins and have an impact on physiological processes. The levels of protein oxidation, nitration and glycation adducts were assayed, using liquid chromatography coupled with tandem mass spectrometry, in total leaf extracts over a diurnal cycle and when exposed to conditions that promote oxidative stress. Changes in the levels of oxidation, glycation and nitration adducts were found between the light and dark phases under non‐stress conditions. A comparison between wild‐type plants and a mutant lacking peptide methionine sulfoxide reductase (pmsr2‐1) showed increased protein oxidation, nitration and glycation of specific amino acid residues during darkness in pmsr2‐1. Short‐term excess light exposure, which promoted oxidative stress, led to increased protein glycation, specifically by glyoxal. This suggested that any increased oxidative damage to proteins was within the repair capacity of the plant. The methods developed here provide the means to simultaneously detect a range of protein oxidation, nitration and glycation adducts within a single sample. Thus, these methods identify a range of biomarkers to monitor a number of distinct biochemical processes that have an impact on the proteome and therefore the physiological state of the plant.
The cell morphogenesis gene SPIRRIG in Arabidopsis encodes a WD/BEACH domain protein
Authors: Saedler Rainer , Jakoby Marc , Marin Birger , Galiana‐Jaime Elena , Hülskamp Martin
Published: April 2009

WD40/BEACH domain proteins have been implicated in membrane trafficking and membrane composition events in Dictyostelium and Drosophila. In this paper, we show that the Arabidopsis SPIRRIG (SPI) gene encodes a WD40/BEACH domain protein. The cellular analysis revealed fragmented vacuoles in root hairs similar to those found in the corresponding Dictyostelium mutants, suggesting a related cellular function. The phenotypic analysis revealed that spi mutants share all phenotypic aspects of mutants in the actin polymerization‐regulating ARP2/3 pathway, including distorted trichomes, less lobing of epidermal pavement cells, disconnected epidermal cells on various organs, and shorter root hairs. This complete phenotypic overlap suggests that this WD40/BEACH domain protein and the actin‐regulating ARP2/3 pathway are involved in similar growth processes.