bluuebunny/crossref_metadata_2025 huggingface dataset:
GeneDesign: Rapid, automated design of multikilobase synthetic genes
Authors: Richardson Sarah M. , Wheelan Sarah J. , Yarrington Robert M. , Boeke Jef D.
Published: February 2006

Modern molecular biology has brought many new tools to the geneticist as well as an exponentially expanding database of genomes and new genes for study. Of particular use in the analysis of these genes is the synthetic gene, a nucleotide sequence designed to the specifications of the investigator. Typically, synthetic genes encode the same product as the gene of interest, but the synthetic nucleotide sequence for that protein may contain modifications affecting expression or base composition. Other desirable changes typically involve the revision of restriction sites. Designing synthetic genes by hand is a time-consuming and error-prone process that may involve several computer programs. We have developed a tools environment that combines many modules to provide a platform for rapid synthetic gene design for multikilobase sequences. We have used GeneDesign to successfully design a synthetic Ty1 element and a large variety of other synthetic sequences. GeneDesign has been implemented as a publicly accessible Web-based resource and can be found at http://slam.bs.jhmi.edu/gd.
The Bible among Lutherans in America: The ELCA as a Test Case
Authors: Heen Erik M.
Published: February 2006

Abstract:  This article describes the biblical hermeneutics that inform the Evangelical Lutheran Church in America by comparing the ELCA's tradition of biblical interpretation with that of the Lutheran Church‐Missouri Synod. It sets both against the great social and intellectual challenges of the early twentieth century, including the modernist/fundamentalist controversy. One commonality that surfaces is that both church bodies appropriated pre‐modern hermeneutical impulses for “counter modern” biblical apologetics. In this process the LC‐MS privileged the period of Lutheran Orthodoxy (17th century) while the ELCA constructed its hermeneutical paradigm through a recovery of the early Reformation (Luther). This observation suggests that both interpretive trajectories need further historical as well as theological review and revision.
Luther's Legacy in American Lutheran Women's Bible Study
Authors: Jacobson Diane Levy , Schersten LaHurd Carol , Wilds McArver Susan
Published: February 2006

Abstract:  Every year thousands of Lutheran lay women explore the Bible through the Women's Bible Study featured in the denomination's monthly women's magazine. This tradition began with the women's missionary societies of the late 19th and early 20th centuries and the recruiting of Lutheran writers for year‐long series. In the ELCA the study is published in the magazine Lutheran Woman Today. The authors have been attentive to biblical critical methods, Lutheran theology, and the lives and faith of people in parishes. Participants report that the women's Bible study contributes to fellowship and to their goals of biblical living and deepened relationships with God, their families, and each other.
Reconstructing Paul Tillich's Anthropology: Multidimensional Life in Dialogue with Feminist Process Theology
Authors: Roesler Tabea
Published: February 2006

Abstract:  Influenced by feminist process theology, this article offers a difference‐oriented reconstruction of Paul Tillich's anthropology. The first section analyzes Tillich's anthropological reductionism at the “basis”(Grund ) of his unity‐oriented binary ontology and epistemology. The second section discloses Tillich's “reverse‐dynamics,” co‐present beside and despite of his anthropological “basis.” This concept of reverse‐dynamics prepares a new, pneumatologically and christologically deepened, “ground”(Grund ) for personhood and life as multidimensional.
Dietrich Bonhoeffer and Asian Contextual Theology in relation to Minjung and Prajna of Inter‐being
Authors: Chung Paul S.
Published: February 2006

Abstract:  Dietrich Bonhoeffer remains an influential figure in inspiring Asian contextual theology of minjung (the poor). In addition to political reading of minjung, a Buddhist wisdom (prajna) offers a basis for understanding of religious dimension of minjung. However, there has been no discussion about Bonhoeffer's legacy of theology after Auschwitz in critical dialogue with Asian contextual theology.
Test‐taking Strategies of 12‐ and 13‐year‐old Hungarian Learners of EFL: Why Whales Have Migraines
Authors: Nikolov Marianne
Published: February 2006

This paper gives an account of a project exploring 12‐ and 13‐year‐old children's uses of strategies while solving reading and writing test tasks in English as a foreign language (EFL). The study was conducted to provide insights into how learners go about solving tasks and what they think and rely on while doing them. The first part provides an overview of strategy research and test‐taking strategy research. The second part provides the background of the project and describes the 52 participants, the data collection instruments, and the procedures. The discussions include four case studies of two high achievers and two low achievers and explore what role individual differences played, what strategies learners applied, and how the findings have contributed to a better understanding of what we mean by test‐taking strategies. This exploratory qualitative study raises several questions. The analyses of the particular strategies and case studies might provide valuable insights into learners’ uses of strategies and useful feedback for test designers and teachers.
Second Language Acquisition of Variable Structures in Spanish by Portuguese Speakers
Authors: Geeslin Kimberly L. , Guijarro‐Fuentes Pedro
Published: February 2006

This study provides a model for examining the second language (L2) acquisition of structures where the first language (L1) and L2 are similar, and where native speaker (NS) use varies. Research on the copula contrast in Spanish (ser and estar mean “to be”) has shown that an assessment of learner choice cannot rely on an error analysis (Geeslin, 2001, 2003a; Geeslin & Guijarro‐Fuentes, 2004). Obligatory contexts are difficult to identify because norms for copula use with adjectives are changing (Silva‐Corvalán, 1994; Guijarro‐Fuentes & Geeslin, in press) and NSs vary in their choices for a given context (Geeslin, 2003a). A more effective approach to assessing the acquisition of variable structures incorporates statistical models that identify features of the discourse that best predict copula choice for NSs and nonnative speakers (NNSs). Our research design, which incorporates semantic and discourse‐pragmatic constraints, is the first to apply this approach to the acquisition of L2 Spanish by Portuguese‐speaking learners. Because Portuguese also possesses copula choice, unlike previously researched L1s, our research also shows that the L2 and the L1 can be compared through analyses based on discourse features and the degree to which each predicts copula choice.

Three participant groups were studied: NSs of Spanish (N = 19), Portuguese‐speaking NNSs of Spanish (N = 11), and NSs of Portuguese (N = 11). Thus, our definition of NS targets comes from two separate sociolinguistic studies in addition to our study of L2 data. The data elicited using a 28‐item contextualized preference task were coded and analyzed according to seven discourse features present in each [copula + adjective] context. Results show that both groups of NSs share the same four predictors, differing only in the level of significance of each. In contrast, the Portuguese NNSs of Spanish are different from each of the other two groups. Our study connects work on the acquisition of Spanish by Portuguese learners to the growing body of research on L2 variation.
Handling Discourse: Gestures, Reference Tracking, and Communication Strategies in Early L2
Authors: Gullberg Marianne
Published: February 2006

The production of cohesive discourse, especially maintained reference, poses problems for early second language (L2) speakers. This paper considers a communicative account of overexplicit L2 discourse by focusing on the interdependence between spoken and gestural cohesion, the latter being expressed by anchoring of referents in gesture space. Specifically, this study investigates whether overexplicit maintained reference in speech (lexical noun phrases [NPs]) and gesture (anaphoric gestures) constitutes an interactional communication strategy. We examine L2 speech and gestures of 16 Dutch learners of French retelling stories to addressees under two visibility conditions. The results indicate that the overexplicit properties of L2 speech are not motivated by interactional strategic concerns. The results for anaphoric gestures are more complex. Although their presence is not interactionally motivated, their spatial articulation is. A learner‐ and processing‐oriented account for both speech and gesture is discussed.
The Effect of Crossing‐Network Trading on Dealer Market's Bid‐Ask Spreads
Authors: Gresse Carole
Published: February 2006

This article provides new insights into market competition between traditional exchanges and alternative trading systems in Europe. It investigates the relationship between the trading activity of a crossing network (CN) and the liquidity of a traditional dealer market (DM) by comparing data from the SEAQ quote‐driven segment of the London Stock Exchange (LSE) and internal data from the POSIT crossing network. A cross‐sectional analysis of bid‐ask spreads shows that DM spreads are negatively related to CN executions. Risk‐sharing benefits from CN trading dominate fragmentation and cream‐skimming costs. Further, risk‐sharing gains are found to be related to dealer trading in the CN.
Diagnostics of age‐graded linguistic behaviour:  The case of the quotative system1
Authors: Buchstaller Isabelle
Published: February 2006

This article presents a cross‐variety investigation of quotatives be like and go in apparent and real time. Distributional and attitudinal evidence points to a change in progress as the underlying process for the distribution of be like. However, there is also evidence of life‐span change (Sankoff to appear). The patterning of go across age is much less clear‐cut. It could be interpreted as age grading or as a change in progress. This paper discusses seemingly contradictory findings from U.S. and British English. It will be suggested that the distribution of go is due to unstable behaviour at both the individual and the community level. Furthermore, there is evidence that go has a latent presence in the linguistic repertoire and was picked up again after its frequency dipped due to the introduction of be like. This finding ties in with other reported cases of recycling of variables (Dubois and Horvath 1999).
The dark arts of good people:  How popular culture negotiates ‘spin’ in NBC's The West Wing1
Authors: Richardson Kay
Published: February 2006

This paper argues that the popular American TV series The West Wing provides, within popular culture, a sympathetic and realistic view of the work of the political ‘spin doctor’. This enterprise is sociolinguistically interesting because of the extensive range of metalinguistic and metacommunicative reflection which it involves. The West Wing centres its storytelling within the world of the spin doctor, thus making such characters central to the narrative: further, it constructs them as likeable, virtuous characters who are also clever. Their cleverness is a matter of sociolinguistic performance expressed in dialogue which displays their professional writing craft skills and their spontaneous witty badinage.
Adoptive transfer of nontransgenic mesenteric lymph node cells induces colitis in athymic HLA-B27 transgenic nude rats
Authors: Hoentjen F , Tonkonogy S L , Liu B , Sartor R B , Taurog J D , Dieleman L A
Published: February 2006

HLA-B27 transgenic (TG) rats develop spontaneous colitis when colonized with intestinal bacteria, whereas athymic nude (rnu/rnu) HLA-B27 TG rats remain disease free. The present study was designed to determine whether or not HLA-B27 expression on T cells is required for development of colitis after transfer of mesenteric lymph node (MLN) cells into rnu/rnu HLA-B27 recipients. Athymic nontransgenic (non-TG) and HLA-B27 TG recipients received MLN cells from either TG or non-TG rnu/+ heterozygous donor rats that contain T cells. HLA-B27 TG rnu/rnu recipients receiving either non-TG or TG MLN cells developed severe colitis and had higher caecal MPO and IL-1β levels, and their MLN cells produced more IFN-γ and less IL-10 after in vitro stimulation with caecal bacterial lysate compared to rnu/rnu non-TG recipients that remained disease free after receiving either TG or non-TG cells. Interestingly, proliferating donor TG T cells were detectable one week after adoptive transfer into rnu/rnu TG recipients but not after transfer into non-TG recipients. T cells from either non-TG or TG donors induce colitis in rnu/rnu TG but not in non-TG rats, suggesting that activation of effector T cells by other cell types that express HLA-B27 is pivotal for the pathogenesis of colitis in this model.
Specific messenger RNA expression for signal transduction molecules by lipopolysaccharide in intestinal macrophages
Authors: Nakata K , Inagawa H , Nishizawa T , Kohchi C , Soma G-I
Published: February 2006

Intestinal macrophages are known to display profound inflammatory anergy in response to lipopolysacchraide (LPS). To study the mechanisms of unresponsiveness of intestinal macrophages to LPS, we compared the mRNA expression of molecules associated with signal transduction of intestinal macrophages with those of other tissue macrophages. Also cellular localization of CD14 protein was examined. Intestinal, alveolar and peritoneal macrophages were isolated from rats or mice. The expression of mRNA was assessed by real-time PCR, and cellular localization of CD14 protein was examined by flow cytometry. Cellular responses to LPS were examined by production of TNF and NO. The expression of CD14 mRNA in intestinal macrophages was lower than for peritoneal macrophages but higher than for alveolar macrophages. The mRNA expression of other molecules corresponding to intracellular signal transduction in intestinal macrophages was similar with alveolar and peritoneal macrophages. Despite the presence of CD14 mRNA, proteins of CD14 were not detected on cell surfaces of intestinal macrophages, and induction of TNF or NO responding to LPS were not detected. Flow cytometric analysis demonstrated that CD14 protein was not expressed on the cell surface but was expressed inside intestinal macrophages. The unresponsiveness of intestinal macrophages after LPS exposure is considered to be largely attributed to the lack of CD14 protein on their cell surfaces. However, CD14 protein was expressed inside of the cells, suggesting that post-transcriptional regulation rather than transcriptional suppression may play a dominant role in determining the phenotype of the intestinal macrophages.
Comparative studies on the roles of mediator molecules in expression of the suppressor activity ofMycobacterium aviumcomplex-induced immunosuppressive macrophages against T cell and B cell mitogenic responses
Authors: Cai S , Shimizu T , Tomioka H
Published: February 2006

Mycobacterium avium complex-induced immunosuppressive macrophages (MAC-MΦs) exhibit suppressor activity against concanavalin A-induced T cell mitogenesis (T cell Con A mitogenesis). We examined the profiles of the MAC-MΦ-mediated suppression of lipopolysaccharide-induced B cell mitogenesis (B cell LPS mitogenesis) and found the following. First, although NG-monomethyl-L-arginine and carboxy-PTIO effectively blocked the MAC-MΦ’s suppressor activity against T cell Con A mitogenesis, MAC-MΦ’s action against B cell LPS mitogenesis was only weakly affected by these NO-reducing agents. Second, B cell LPS mitogenesis was remarkably more susceptible to MAC-MΦ-derived reactive oxygen intermediates than T cell Con A mitogenesis. Third, B cell LPS mitogenesis was less susceptible to the inhibitory effects of the other MAC-MΦ-derived suppressor mediators, including free fatty acids, TGF-β and prostaglandin E2, than T cell Con A mitogenesis. Fourth, MAC-MΦ’s suppressor activity was strongly dependent on B7-1 like molecule-mediated cell contact with target cells only in the case of T cell Con A mitogenesis. Therefore, there are significant differences in the modes of suppressor action of MAC-MΦs against T cell and B cell mitogenesis.
T cell-mediated oral tolerance is intact in germ-free mice
Authors: Walton K L W , Galanko J A , Balfour Sartor R , Fisher N C
Published: February 2006

Commensal enteric bacteria stimulate innate immune cells and increase numbers of lamina propria and mesenteric lymph node (MLN) T and B lymphocytes. However, the influence of luminal bacteria on acquired immune function is not understood fully. We investigated the effects of intestinal bacterial colonization on T cell tolerogenic responses to oral antigen compared to systemic immunization. Lymphocytes specific for ovalbumin–T cell receptor (OVA–TCR Tg+) were transplanted into germ-free (GF) or specific pathogen-free (SPF) BALB/c mice. Recipient mice were fed OVA or immunized subcutaneously with OVA peptide (323–339) in complete Freund’s adjuvant (CFA). Although the efficiency of transfer was less in GF recipients, similar proportions of cells from draining peripheral lymph node (LN) or MLN were proliferating 3–4 days later in vivo in GF and SPF mice. In separate experiments, mice were fed tolerogenic doses of OVA and then challenged with an immunogenic dose of OVA 4 days later. Ten days after immunization, lymphocytes were restimulated with OVA in vitro to assess antigen-specific proliferative responses. At both high and low doses of OVA, cells from both SPF and GF mice fed OVA prior to immunization had decreased proliferation compared to cells from control SPF or GF mice. In addition, secretion of interferon (IFN)-γ and interleukin (IL)-10 by OVA–TCR Tg+ lymphocytes was reduced in both SPF and GF mice fed OVA compared to control SPF or GF mice. Unlike previous reports indicating defective humoral responses to oral antigen in GF mice, our results indicate that commensal enteric bacteria do not enhance the induction of acquired, antigen-specific T cell tolerance to oral OVA.
Association of rheumatoid factor production with FcγRIIIa polymorphism in Taiwanese rheumatoid arthritis
Authors: Chen J-Y , Wang C-M , Wu J-M , Ho H-H , Luo S-F
Published: February 2006

Fcγ receptors (FcγR) impact upon the development of inflammatory arthritis through immune complex stimulation and proinflammatory cytokine production. FcγRIIa, FcγRΙΙΙa and FcRγIIIb polymorphisms were genotyped in 212 rheumatoid arthritis (RA) patients and 371 healthy control subjects using an allelic-specific polymerase chain reaction (PCR). No significant skewing in the distribution of FcγRIIa H/R131, FcγRIIIa F/V158 and FcγRIIIb NA1/NA2 was found between RA patients and healthy control subjects. However, a significant skewing distribution of the FcγRIIIa F/V158 polymorphism was observed between rheumatoid factor (RF)-positive versus RF-negative RA patients (P = 0·01). The low-affinity FcγRIIIa F158 allele seems to have a protective role in RF production, in comparison with the FcγRIIIa V158 allele (P = 0·004; OR = 0·485; 95% CI: 0·293–0·803). A high frequency of FcγRIIIa F/F158 was identified in RA patients with negative RF compared with RF-positive patients (for FF158 versus FV158 + VV158; P = 0·002; OR = 0·372; 95% CI: 0·194–0·713). In addition, no association was found between FcγRIIa H/R131, FcγΡIIIa F/V158 and FcγRIIIb NA1/NA2 polymorphisms and other clinical parameters. The results of this study suggest that three activating FcγRs polymorphisms lack association with RA but FcγIIIa F/V158 polymorphism may influence RF production and IgG RF immune complex handling in Taiwanese RA patients.
CD127 expression and regulation are altered in the memory CD8 T cells of HIV-infected patients − reversal by highly active anti-retroviral therapy (HAART)
Authors: Colle J-H , Moreau J-L , Fontanet A , Lambotte O , Joussemet M , Delfraissy J-F , Thèze J
Published: February 2006

HIV infection activates abnormally the immune system and the chronic phase is accompanied by marked alterations in the CD8 compartment. The expression of CD127 (IL-7R alpha chain) by memory CD8 T lymphocytes in HIV-infected patients is analysed and reported. The memory CD8 T cell subset was characterized by expression of CD45RA and CD27 markers, and CD127 cell surface expression was measured ex vivo by four-colour flow cytometry. HIV infection was associated with a fall in the proportion of CD127+ cells among memory CD8 lymphocytes that resulted in a higher CD127– CD45RA–CD27+ CD8 T cell count in HIV-infected patients. Diminished CD127 cell surface expression [mean fluorescence intensity (MFI)] by positive cells was also observed in this subset. The data suggest that these defects were reversed by highly active anti-retroviral therapy (HAART). The regulation of CD127 expression was also studied in vitro. Down-regulation of CD127 by interkeukin (IL)-7 was observed in memory CD8 lymphocytes from healthy donors and HAART patients. Expression of CD127 by memory CD8 lymphocytes cultured in the absence of IL-7 confirmed that IL-7R regulation is altered in viraemic patients. Under the same experimental conditions, memory CD8 lymphocytes from HAART patients were shown to express CD127 at levels comparable to cells from healthy individuals. Altered CD127 cell surface expression and defective CD127 regulation in the memory CD8 T lymphocytes of HIV-infected patients are potential mechanisms by which these cells may be impeded in their physiological response to endogenous IL-7 stimulatory signals. Our data suggest that these defects are reversed during the immune reconstitution that follows HAART.
Increased expression and secretion of interleukin-6 in human parvovirus B19 non-structural protein (NS1) transfected COS-7 epithelial cells
Authors: Hsu T-C , Tzang B-S , Huang C-N , Lee Y-J , Liu G-Y , Chen M-C , Tsay G J
Published: February 2006

Human parvovirus B19 (B19) has been associated with a variety of autoimmune diseases, including rheumatoid arthritis (RA) and systemic lupus erythematosus (SLE). We have demonstrated previously that B19 non-structural protein (NS1) induced apoptosis through the mitochondria cell death pathway in COS-7 epithelial cells and that B19 NS1 may play a role in the pathogenesis of autoimmune diseases. In order to examine the expression profiles of cytokines and chemokines in B19 NS1 transfected COS-7 cells, we constructed the NS1 gene in the pEGFP-C1 vector named enhanced green fluorescence protein gene (EGFP)-NS1. COS-7 cells were transfected with EGFP or EGFP-NS1 plasmid. The expression profiles of cytokines and chemokines, including interleukin (IL)-1β, IL-5, IL-6, IL-8, IL-10, tumour necrosis factor (TNF)-α, transforming growth factor (TGF)-β, granulocyte–macrophage colony-stimulating factor (GM-CSF), growth-related oncogene α (GROα), interferon gamma-inducible protein (IP)-10, stromal cell derived factor (SDF)-1, macrophage inflammatory protein (MIP)-1β, monocyte chemoattractant protein (MCP)-1, regulated upon activation normal T cell expressed and secreted (RANTES), Fractalkine, CX3CR1, CCR2, CCR5 and CCR11 were examined in COS-7 cells, EGFP and EGFP-NS1 transfected cells using enzyme-linked immunosorbent assay (ELISA) or reverse transcription–polymerase chain reaction (RT–PCR). Increased expression and levels of IL-6 were found in EGFP-NS1 transfected cells using RT–PCR and ELISA. There were no significant increases in the expression of IL-1β, IL-8, IP-10, SDF-1, RANTES, Fractalkine, CX3CR-1, CCR2, CCR5, CCR11, TNF-α, GM-CSF and TGF-β using RT–PCR. There were no significantly increased levels of IL-5, IL-10, TNF-α, TGF-β, GROα, MIP-1β and MCP-1 found by ELISA in this study. Our results show that increased expression and secretion of IL-6 in B19 NS1 transfected epithelial cells may play a role in the pathogenesis of autoimmune diseases.
A role for CD4+CD25+ T cells in regulation of the immune response during human tuberculosis
Authors: Ribeiro-Rodrigues R , Resende Co T , Rojas R , Toossi Z , Dietze R , Boom W H , Maciel E , Hirsch C S
Published: February 2006

Active tuberculosis (TB) is associated with prolonged suppression of Mycobacterium tuberculosis (MTB)-specific immune responses, but mechanisms involved are understood incompletely. We investigated a potential role for CD4+CD25+ regulatory T cells in depressed anti-MTB immunity by evaluating serially CD4 cell phenotype and interferon (IFN)-γ production by mononuclear cells from patients with TB. At diagnosis, frequencies of CD4+CD25+ T cells were increased in blood from TB patients compared to healthy purified protein derivative (PPD)-positive controls (with a history of prior TB exposure), and remained elevated at completion of therapy (6 months). By contrast, expression of another activation marker, CD69, by CD4 T cells was increased at diagnosis, but declined rapidly to control levels with treatment. Among CD4+CD25+ T cells from TB patients at diagnosis those expressing high levels of CD25, probably representing regulatory T cells, were increased 2·9-fold when compared to control subjects, while MTB-stimulated IFN-γ levels in whole blood supernatants were depressed. A role for CD4+CD25+ T cells in depressed IFN-γ production during TB was substantiated in depletion experiments, where CD25+-depleted CD4 T cells produced increased amounts of IFN-γ upon MTB stimulation compared to unseparated T cells. At follow-up, IFN-γ production improved most significantly in blood from TB patients with high baseline frequencies of CD4+CD25+ T cells (more than threefold higher than controls for both total and CD25hi+ CD4 T cells), who also had a significant drop in frequencies of both total and ‘regulatory’ CD4+CD25+ T cells in response to treatment. Expansion of CD4+CD25+ regulatory T cells during active TB may play a role in depressed T cell IFN-γ production.
Balance of inflammatory response in stable gingivitis and progressive periodontitis lesions
Authors: Honda T , Domon H , Okui T , Kajita K , Amanuma R , Yamazaki K
Published: February 2006

The balance between inflammatory mediators and their counter-regulatory molecules may be crucial for determining the outcome of immune pathology of periodontal diseases. Based on clinical and immunological findings, the immune response in stable gingivitis lesion is supposed to be in balance, whereas the response is skewed towards the predominance of proinflammatory reactivity in progressive periodontitis lesion. However, this hypothesis has not been verified. Therefore, the aim of this study was to compare the gene expression profile of inflammatory mediators including proinflammatory cytokines and other inflammatory molecules, and anti-inflammatory cytokines by using quantitative real-time polymerase chain reaction in gingivitis and periodontitis lesions showing distinct clinical entities. For inflammatory mediators, interleukin (IL)-1β, interferon (IFN)-γ and receptor activator of nuclear factor (NF)-κB ligand tended to be higher in periodontitis, whereas tumour necrosis factor (TNF)-α and IL-12 p40 showed no difference. Heat-shock protein 60 (HSP60) expression was up-regulated significantly in periodontitis. For anti-inflammatory cytokines, transforming growth factor (TGF)-β1 expression tended to be higher in periodontitis compared with gingivitis, whereas no difference was observed for IL-10 and IL-4. These findings support further our previous finding that autoimmune response to HSP60 may exert in periodontitis lesion, and suggest that perhaps subtle differences in the balance of cytokines may result in different disease expression.
IA-2 antibody isotypes and epitope specificity during the prediabetic process in children with HLA-conferred susceptibility to type I diabetes
Authors: Hoppu S , Härkönen T , Ronkainen M S , Simell S , Hekkala A , Toivonen A , Ilonen J , Simell O , Knip M
Published: February 2006

The natural history of preclinical diabetes is partly characterized, but there is still limited information on the dynamics of the immune response to β-cell autoantigens during the course of preclinical disease. The aim of this work was to assess the maturation of the humoral immune response to the protein tyrosine phosphatase(PTP)-related proteins (IA-2 and IA-2β) in preclinical type I diabetes (TID). Forty-five children participating in the Finnish Type I Diabetes Prediction and Prevention (DIPP) Study who had seroconverted to IA-2 antibody positivity were analysed. Specific radiobinding assays were used to determine IA-2/IA-2β epitope-specific antibodies (the juxtamembrane (JM) region of IA-2, PTP-like domain and βPTP-like domain) and isotype-specific IA-2 antibodies. Individual areas under the curve (AUC) over the observation period were calculated for total IA-2 antibodies, each isotype and specific epitope responses. The children who progressed to TID tended to have an initial IA-2 JM epitope response more frequently (P = 0·06), and this response was more often dominant during the observation period (P < 0·05). The children who did not progress to TID had IgE-IA-2 more frequently (70%; versus progressors 27%; P < 0·05), and had higher integrated titres of IgE-IA-2 antibodies (P < 0·05). The occurrence of IgE-IA-2 antibodies was protective even when combined with positivity for IA-2 JM antibodies (P = 0·002). IgE-IA-2 antibody reactivity may be a marker of a regulatory immune response providing protection against or delaying progression to TID among IA-2 antibody-positive young children with HLA-conferred disease susceptibility.
A novel model of acute liver injury in mice induced by T cell-mediated immune response to lactosylated bovine serum albumin
Authors: Xu Ling , Zhao Ying , Qin Yu , Xu Qiang
Published: February 2006

The present study aimed to establish a murine liver injury model with T cell-mediated immune response to mimic the cellular immunological pathogenesis of hepatitis. Bovine serum albumin was conjugated with lactose by redox to prepare lactosylated bovine serum albumin (LacBSA). The liver injury was induced in C57BL/6 mice by a T cell-mediated immune response to LacBSA. After a systemic injection of LacBSA, which elicited immune response in mice sensitized twice with LacBSA, liver injury was observed with obvious increase of aminotransferase levels in serum and hepatocelluar damage in liver histology. However, the splenectomy before the elicitation significantly alleviated the liver injury. Direct contact between spleen T cells/nonparenchymal cells and hepatocytes was proved to be essential to induce the release of alanine aminotransferase in the culture supernatant. In addition, both levels of Fas mRNA in liver tissues and Fas Ligand in spleen cells were up-regulated at 6 h after the elicitation. The Fas mRNA expression was maintained in a relatively high level at 18 h while Fas Ligand began to decrease after 12 h. These results demonstrated that the T cell immune response to LacBSA could be successfully localized in the liver to induce an immunological liver injury in mice. The pathogenesis of the liver injury might involve the interaction between the liver-infiltrating lymphocytes and hepatocytes through Fas/FasL pathway.
Selection of Epstein-Barr virus specific cytotoxic T lymphocytes can be performed with B lymphoblastoid cell lines created in serum-free media
Authors: Gallot G , Vollant S , Vivien R , Clémenceau B , Ferrand C , Tiberghien P , Gaschet J , Robillard N , Vié H
Published: February 2006

Epstein-Barr Virus (EBV)-transformed B lymphoblastoid cell lines (BLCL) are currently used for numerous applications in cellular immunology. Where protocols destined for clinical application are concerned, the final choice of assay is made according to a risk/benefit ratio analysis. In this balance the use of xenogenic or allogenic serum has always been a major concern, as it carries both an infectious and an immunological risk. So far, it is unknown whether serum can be omitted from the entire BLCL selection procedure. In addition, as BLCL have been described as heterogeneous, serum deprivation may affect their antigen-presenting capacity. In the present study, BLCL were generated in the absence or presence of fetal calf serum (referred to as BLCL0 or BLCLFCS, respectively). Next, in order to assess the antigen-presenting capacity of these cells, we compared the ability of BLCL0 and BLCLFCS cells to stimulate the EBV-specific repertoire of the corresponding donor’s peripheral blood mononuclear cells in vitro. Our results showed that addition of serum was not essential for BLCL infection and culture, and that as far as we could determine, BLCL0 cells were as effective as BLCLFCS in reactivating the EBV-specific T-cell repertoire in vitro. Notably, FCS-specific T-lymphocytes can be detected among the BLCLFCS-specific CD4+-CTL. Not only was this latter observation unexpected for an EBV-seropositive donor, but it implied that the BLCL had captured and processed the corresponding FCS-derived solubles antigens; taken together our results emphasized the interest of the possibility to generate BLCL0, both for research and for clinical applications.
Maturational alterations of peripheral T cell subsets and cytokine gene expression in 22q11.2 deletion syndrome
Authors: Kanaya Y , Ohga S , Ikeda K , Furuno K , Ohno T , Takada H , Kinukawa N , Hara T
Published: February 2006

Chromosome 22q11.2 deletion syndrome is a common disorder characterized by thymic hypoplasia, conotruncal cardiac defect and hypoparathyroidism. Patients have a risk of infections and autoimmunity associated with T lymphocytopenia. To assess the immunological constitution of patients, the numerical changes and cytokine profile of circulating T cells were analysed by flow cytometry and real-time polymerase chain reaction (PCR). CD3+, CD4+, T cell receptor (TCR)αβ+ or CD8αα+ cell counts were lower, and CD56+ cell counts were higher in patients than in controls during the period from birth to adulthood. The ageing decline of CD3+ or CD4+ cell counts was slower in patients than in controls. The proportion of CD8αα+ cells increased in controls, and the slope index was larger than in patients. On the other hand, both the number and proportion of Vα24+ cells increased in patients, and the slope indexes tended to be larger than in controls. The positive correlation of the number of T cells with CD8αα+ cells was observed only in patients, and that with Vα24+ cells was seen only in controls. No gene expression levels of interferon (IFN)-γ, interleukin (IL)-10, transforming growth factor (TGF)-β, cytotoxic T lymphocyte antigen 4 (CTLA4) or forkhead box p3 (Foxp3) in T cells differed between patients and controls. There was no significant association between the lymphocyte subsets or gene expression levels and clinical phenotype including the types of cardiac disease, hypocalcaemia and frequency of infection. These results indicated that T-lymphocytopenia in 22q11.2 deletion patients became less severe with age under the altered composition of minor subsets. The balanced cytokine profile in the limited T cell pool may represent a T cell homeostasis in thymic deficiency syndrome.
Fatal central venous catheter‐related infection in haemophilia
Authors: CRARY S. E. , BUCHANAN G. R. , JOURNEYCAKE J. M.
Published: February 2006

Summary.  Central venous catheters (CVC) are frequently used in children with haemophilia to deliver factor infusions for the treatment or prophylaxis of bleeding. Complications of CVCs in patients with haemophilia include thrombosis and infection. We report a young boy with severe haemophilia A and an inhibitor who developed disseminated Staphylococcus aureus infection most likely related to a CVC. To our knowledge, this is the first reported case of fatal sepsis secondary to a CVC in a patient with haemophilia.
Occurrence of inhibitors in previously untreated or minimally treated patients with haemophilia A after exposure to a plasma‐derived solvent‐detergent factor VIII concentrate
Authors: GRINGERI A. , MONZINI M. , TAGARIELLO G. , SCARAGGI F. A. , MANNUCCI P. M. ,   THE EMOCLOT15 STUDY MEMBERS
Published: February 2006

Summary.  A solvent‐detergent virus‐inactivated plasma‐derived FVIII concentrate (SD‐pdFVIII) has been employed for treatment of Italian patients with haemophilia A for 15 years. This product is a non‐monoclonally purified, high purity FVIII concentrate, containing large amounts of von Willebrand factor (VWF). A retrospective survey was carried out in Italy in order to evaluate the immunogenicity of SD‐pdFVIII in previously untreated patients (PUPs) or in minimally treated patients (MTPs), i.e. previously exposed for up to 5 days only to other plasma‐derived concentrates. The survey included 99 patients with ages ranging from 6 to 64 years (median = 21.3) of whom 31 PUPs and 68 MTPs, the latter with a median of four exposure days (EDs; range 1–5) to other plasma products. Surveyed patients had been exposed to SD‐pdFVIII for a median of 83 EDs (range 21–1580). Seven patients (three PUPs and four MTPs), all with severe haemophilia, had developed inhibitors [7.1%, 95%; confidence interval: 3–14%] after a median of 11 EDs (range 4–22). Of them, two were low responders (≤5 BU mL−1) and five high responders. Two low responders and one high responder tolerated inhibitors spontaneously; one high responder underwent immunotolerance treatment with complete, long‐standing response. These findings show that SD‐pdFVIII is at low risk of inhibitor development in PUPs and MTPs with severe and moderate haemophilia, the risk of inhibitor development being similar to that previously reported for other plasma‐derived VWF‐containing FVIII products.
Morphology of the bone marrow after stem cell transplantation
Authors: van Marion A M W , Thiele J , Kvasnicka H M , van den Tweel J G
Published: February 2006

In many haematological conditions the only curative option is stem cell (SCT) or bone marrow (BM) transplantation. Little information exists about BM morphology following non‐ablative engraftment. During the pretransplantation period and depending on the kind of pretreatment, there may be hypoplasia, residual disease and varying degrees of fibrosis. In the post‐transplantation period, after 1–3 weeks of transfusion‐dependent pancytopenia, the first signs of successful engraftment are indicated by the recurrence of neutrophils, monocytes and erythrocytes in the peripheral blood. In the BM there is slow regeneration of erythropoiesis, followed by the other lineages of haematopoiesis and increase in reticulin fibres or even a resolution of fibrosis. Diagnostic problems arise when neoplastic lympho‐ or haematopoiesis are maintained following transplantation. Moreover, there may be a significant graft versus tumour response reaction or an already relapsing disease needing aggressive treatment. On the other hand, a conspicuous dyshaematopoiesis should not be mistaken as representing a myelodysplastic syndrome. The presence of granulomas being treatment‐related or a manifestation of intercurrent granulomatous disease has to be considered. More advanced knowledge of the histological features of regenerating BM will certainly aid the recognition of relapsing disease and is needed for the adequate reporting of post‐transplant alterations associated with a successful or failing engraftment.
Morphometrical differences between pseudo‐epitheliomatous hyperplasia in granular cell tumours and squamous cell carcinomas
Authors: Abu‐Eid R , Landini G
Published: February 2006

Aim : Granular cell tumour (GCT) is a benign tumour which occasionally induces pseudo‐epitheliomatous hyperplasia (PEH) of the covering epithelium, mimicking squamous cell carcinoma (SCC) invasion. PEH cells do not show marked atypia, but the presence of pseudoinvasive patterns remains a diagnostic problem since several misdiagnosed cases have been reported. This study investigated objective morphometric criteria to distinguish GCT‐PEH from SCC.

Methods and results : The complexity of the epithelial connective tissue interface (ECTI) in 57 profiles from nine SCC and 12 GCT‐PEH cases was analysed using fractal geometry. Epithelial and non‐epithelial cells were segmented using a space partition procedure and analysed morphometrically. GCT ECTI profiles were significantly more complex than those of SCC. The complexity of ECTI quantified by global and local fractal dimensions allowed up to 86% correct discrimination between GCT‐PEH and SCC. The cell‐wise discrimination between the two entities using cellular morphology was 76% but when the two approaches were combined, the correct discrimination was 96%.

Conclusions : The architectural features of GCT‐PEH and SCC show differences which, when quantified, could be used to differentiate the two diagnostic classes. Characterization of these differences may help to elucidate some of the mechanisms of tumour infiltration and metastasis.
Development and characterization of monoclonal antibodies for the immunohistochemical detection of glycodelin A in decidual, endometrial and gynaecological tumour tissues
Authors: Jeschke U , Kuhn C , Mylonas I , Schulze S , Friese K , Mayr D , Speer R , Briese V , Richter D‐U , Haase M , Karsten U
Published: February 2006

Aims : Glycodelin is a glycoprotein with a molecular weight of 28 kDa. Unusual LacdiNAc structures have been identified on glycodelin A, isolated from amniotic fluid. Three major functions of this glycoprotein have been identified. Glycodelin is an immunosuppressive molecule, a marker of morphological differentiation, and a contraceptive. Because no monoclonal antibodies for glycodelin A are commercially available, our aim was to develop and characterize three monoclonal antibodies against this glycoprotein.

Methods and results : Glycodelin A was purified from amniotic fluid by three chromatographic steps and its purity was checked by SDS–PAGE. Antibodies were generated from immunized BALB/c mice. Three IgG1 monoclonal antibodies detecting glycodelin A were cloned. All three antibodies recognized carbohydrate structures of glycodelin A and did not cross‐react with glycodelin S. They are applicable to immunohistochemistry (frozen and paraffin sections), ELISA and Western blots.

Conclusion : The new antibodies can be used for the detection of glycodelin A in frozen and paraffin‐embedded decidual and endometrial tissue. One antibody (A87‐B/D2) can be used for the detection of glycodelin in endometrial and ovarian tumour tissues. Because glycodelin A is a major secretory endometrial product during the luteal phase, in early pregnancy and in gynaecological tumours, the new antibodies are, potentially, valuable tools for the study of endometrial development and tumour progression.
Infiltrative myelopathy by paracoccidioidomycosis. A review and report of nine cases with emphasis on bone marrow morphology
Authors: Resende L S R , Mendes R P , Bacchi M M , Marques S A , Barraviera B , Souza L R , Meira D A , Niéro‐Melo L
Published: February 2006

Aims : To report nine additional well‐defined cases with infiltrative myelopathy by paracoccidioidomycosis (PCM), to describe the specific lesions and infection‐related stromal abnormalities, to review the literature on this type of involvement and to introduce a new cause of granulomatous lesions of bone marrow.

Methods and results : Different bone marrow specimens were studied (aspirated smears, aspirated clots, biopsy imprints and biopsies) from nine patients with acute or subacute forms of PCM known to have PCM infiltrative myelopathy.

Conlusions : The biopsy specimens were the best for demonstrating bone marrow involvement by PCM. The lesions varied from compact and focal granulomas with few fungal cells to numerous disseminated fungal cells within a loose granulomatous inflammatory reaction, with a continuum between these extremes suggesting a spectrum of immune response to the fungi. Other findings such as bone marrow fibrosis, parenchymal coagulative necrosis and bone necrosis were also observed in the affected areas.
Spectrum of pathological changes in both ventricles of patients dying suddenly with arrhythmogenic right ventricular dysplasia. Relation of changes to age
Authors: Fletcher A , Ho S Y , McCarthy K P , Sheppard M N
Published: February 2006

Aims : To quantify the variation in fibrosis, fat and muscle within the walls of both ventricles and within the different regions of the heart from six patients dying suddenly of arrhythmogenic right ventricular dysplasia (ARVD) aged 20–60 years.

Methods : Seven heart regions were examined both macroscopically and histologically using the Picro‐Sirius red stain. Quantification of fibrosis, fat and muscle was performed in each region and transmural layer using grid counting.

Results : There were macroscopic changes in all examined hearts. A higher percentage of fat with less fibrosis and muscle was observed within the right ventricle of the older patients. The left ventricle had more pathology in the older age group. Statistical differences in pathology in the heart were found. Fat predominated in the epicardial layer in the right and left ventricles of all patients, while the interventricular septum was the least affected.

Conclusions : In ARVD, the pathology varies with age in both ventricles, fibrosis being the earliest hallmark of disease, with fatty infiltration evolving later. It should be labelled arrhythmogenic ventricular dysplasia because of biventricular involvement. Histopathologists should therefore sample from whole slices of the heart, so that all the changes can be observed.
Fibroblastic polyp of the colon: clinicopathological analysis of 10 cases with emphasis on its common association with serrated crypts
Authors: Groisman G M , Polak‐Charcon S , Appelman H D
Published: February 2006

Aims : To describe the clinical and pathological features of 10 further cases of fibroblastic polyps (FP), a recently described, distinctive type of colorectal mucosal polyp.

Methods and results : The patients were seven women and three men with ages ranging from 44 to 63 years. The lesions ranged in size from 2 to 4 mm. Eight of the polyps were located in the sigmoid colon. Five cases were associated with hyperplastic polyps. Histologically, FP displayed bland, plump spindle cells with oval nuclei arranged as bundles parallel to the surface or as haphazardly orientated sheets with a focal periglandular or perivascular concentric arrangement. Eight polyps represented mixed fibroblastic/hyperplastic polyps as they contained serrated (hyperplastic) crypts. Immunohistochemically, all cases were positive for vimentin and negative for desmin, smooth‐muscle actin, h‐caldesmon, S100 protein, c‐Kit, epithelial membrane antigen, cytokeratin AE1/3, CD34, CD68, COX‐2, and factor XIIIa. Ultrastructural examination supported the fibroblastic nature of the tumour cells.

Conclusions : FP is a distinctive type of benign mucosal colorectal polyp characterized by its distal location, small size, frequent association with hyperplastic polyps, distinct morphological appearance and typical immunonegativity for markers of specific differentiation. FP with serrated crypts (mixed fibroblastic/hyperplastic polyp) represents a frequent variant of this lesion. Pathologists should recognize FP and discriminate it from other types of colorectal polyps.