bluuebunny/crossref_metadata_2025 huggingface dataset:
THE IMPACT OF CLUSTERIZATION ON THE DEVELOPMENT OF SMALL AND MEDIUM‐SIZED ENTERPRISE (SME) SECTOR
Authors: Navickas Valentinas , Malakauskaitė Asta
Published: September 2009

The impact of clusterization on the development of SME sector has been analysed in this study. The cooperation of companies at national level and on a global scale is becoming more and more important as a tool of economic development. Companies tend to work together in order to share their competencies, reduce various costs, consolidate limited resources, and hereby increase their productivity, innovativeness, and profitability. It must be emphasized that the role of clusterization is crucial in the development of SME sector, as small and medium‐sized enterprises may benefit from economies of scale and extend the operation limits (size‐related limitations of operation are characteristic of most small businesses). Clusters (and similar forms of interorganizational structures) create the environment for innovation and technological advancement. Therefore, small and medium‐sized enterprises may gain additional benefits that include know‐how, cost‐saving options, innovative solutions, etc. The authors of this scientific study have concluded that the competitiveness of SME sector is closely related to the spread and extent of clusterization processes.
Role of TL1A in the Pathogenesis of Rheumatoid Arthritis
Authors: Zhang Jun , Wang Xuehai , Fahmi Hassan , Wojcik Susan , Fikes James , Yu Youhua , Wu Jiangping , Luo Hongyu
Published: September 2009

TNF-like ligand 1A (TL1A), a member of the TNF superfamily, is the ligand of DR3 and DcR3. Several types of cells, such as endothelial cells, monocytes/macrophages, dendritic cells, and CD4 and CD8 T cells, are capable of producing this cytokine. In present study, we demonstrated that TL1A aggravated collagen-induced arthritis in mice. It increased collagen-induced arthritis penetrance and clinical scores as well as the severity of the pathological findings. TL1A administration led to the occurrence of multiple enlarged germinal centers in the spleen, and it boosted serum anti-collagen Ab titers in vivo. In vitro, TL1A augmented TNF-α production by T cells upon TCR ligation, and it greatly enhanced Th17 differentiation and IL-17 production. We further showed that human rheumatoid arthritis (RA) synovial fluids had elevated TL1A titers, and human chrondrocytes and synovial fibroblasts were capable of secreting TL1A upon TNF-α or IL-1β stimulation. Taken together, these data suggest that TL1A secretion in lymphoid organs might contribute to RA initiation by promoting autoantibody production, and TL1A secretion stimulated by inflammatory cytokines in RA joints might be a part of a vicious circle that aggravates RA pathogenesis.
The Effects of Host Age on Follicular Dendritic Cell Status Dramatically Impair Scrapie Agent Neuroinvasion in Aged Mice
Authors: Brown Karen L. , Wathne Gwennaelle J. , Sales Jill , Bruce Moira E. , Mabbott Neil A.
Published: September 2009

Following peripheral exposure, many transmissible spongiform encephalopathy (TSE) agents accumulate first in lymphoid tissues before spreading to the CNS (termed neuroinvasion) where they cause neurodegeneration. Early TSE agent accumulation upon follicular dendritic cells (FDCs) in lymphoid follicles appears critical for efficient neuroinvasion. Most clinical cases of variant Creutzfeldt-Jakob disease have occurred in young adults, although the reasons behind this apparent age-related susceptibility are uncertain. Host age has a significant influence on immune function. As FDC status and immune complex trapping is reduced in aged mice (600 days old), we hypothesized that this aging-related decline in FDC function might impair TSE pathogenesis. We show that coincident with the effects of host age on FDC status, the early TSE agent accumulation in the spleens of aged mice was significantly impaired. Furthermore, following peripheral exposure, none of the aged mice developed clinical TSE disease during their lifespans, although most mice displayed histopathological signs of TSE disease in their brains. Our data imply that the reduced status of FDCs in aged mice significantly impairs the early TSE agent accumulation in lymphoid tissues and subsequent neuroinvasion. Furthermore, the inefficient neuroinvasion in aged individuals may lead to significant levels of subclinical TSE disease in the population.
Long-Lived Colitogenic CD4+ Memory T Cells Residing Outside the Intestine Participate in the Perpetuation of Chronic Colitis
Authors: Nemoto Yasuhiro , Kanai Takanori , Kameyama Kaori , Shinohara Tamako , Sakamoto Naoya , Totsuka Teruji , Okamoto Ryuichi , Tsuchiya Kiichiro , Nakamura Tetsuya , Sudo Tetsuo , Matsumoto Satoshi , Watanabe Mamoru
Published: September 2009

To understand the perpetuation of inflammatory bowel disease (IBD), it is important to clarify whether the colitogenic CD4+ T cells are self-limited effector or long-lived memory T cells. We here investigate the latency of colitogenic CD4+ T cells in the remission stage of colitis under germfree (GF) conditions. We isolated splenic (SP) CD4+ T cells from colitic CD4+CD45RBhigh T cell-injected SCID mice maintained under specific pathogen-free (SPF) conditions and transferred them into SPF or GF SCID mice. Donor colitic SP CD4+ T cells have a characteristic CD44highCD62L−IL-7Rαhigh effector-memory T-type phenotype. Six weeks after transfer of cells to GF SCID mice, one group of mice was continued in GF conditions (GF→GF), and the other was transferred into SPF conditions (GF→SPF). GF→SPF but not GF→GF SCID mice developed colitis with elevated production of Th1 and Th17 cytokines at 4 wk after transfer. Surprisingly, a large number of CD4+ effector-memory T cells and a small but substantial number of central-memory T cells remained resident in SP and bone marrow, but not in lamina propria, of the GF→GF SCID recipients. Consistent with this, GF→SPF but not GF→GF SCID mice rapidly developed colitis. Taken together, these findings suggest that long-lived colitogenic memory CD4+ cells can be established even in the presence of commensal Ags, reside outside the intestine in the absence of commensal bacteria, and participate in the perpetuation of colitis. Thus, blocking a stimulus of colitogenic memory CD4+ cells such as IL-7 may have therapeutic benefit for treatment of inflammatory bowel disease.
Essential Impact of NF-κB Signaling on the H5N1 Influenza A Virus-Induced Transcriptome
Authors: Schmolke Mirco , Viemann Dorothee , Roth Johannes , Ludwig Stephan
Published: September 2009

Systemic infections of humans and birds with highly pathogenic avian influenza A viruses of the H5N1 subtype are characterized by inner bleedings and a massive overproduction of cytokines known as cytokine storm. Growing evidence supports the role of endothelial cells in these processes. The aim of this study was to elucidate determinants of this strong response in endothelial cells with a focus on the transcription factor NF-κB. This factor is known as a major regulator of inflammatory response; however, its role in influenza virus replication and virus-induced immune responses is controversially discussed. By global mRNA profiling of infected cells in the presence or absence of a dominant negative mutant of IκB kinase 2 that specifically blocks the pathway, we could show that almost all H5N1 virus-induced genes depend on functional NF-κB signaling. In particular, activation of NF-κB is a bottleneck for the expression of IFN-β and thus influences the expression of IFN-dependent genes indirectly in the primary innate immune response against H5N1 influenza virus. Control experiments with a low pathogenic influenza strain revealed a much weaker and less NF-κB-dependent host cell response.
Mitochondrial Dysfunction Increases Allergic Airway Inflammation
Authors: Aguilera-Aguirre Leopoldo , Bacsi Attila , Saavedra-Molina Alfredo , Kurosky Alexander , Sur Sanjiv , Boldogh Istvan
Published: September 2009

The prevalence of allergies and asthma among the world’s population has been steadily increasing due to environmental factors. It has been described that exposure to ozone, diesel exhaust particles, or tobacco smoke exacerbates allergic inflammation in the lungs. These environmental oxidants increase the levels of cellular reactive oxygen species (ROS) and induce mitochondrial dysfunction in the airway epithelium. In this study, we investigated the involvement of preexisting mitochondrial dysfunction in the exacerbation of allergic airway inflammation. After cellular oxidative insult induced by ragweed pollen extract (RWE) exposure, we have identified nine oxidatively damaged mitochondrial respiratory chain-complex and associated proteins. Out of these, the ubiquinol-cytochrome c reductase core II protein (UQCRC2) was found to be implicated in mitochondrial ROS generation from respiratory complex III. Mitochondrial dysfunction induced by deficiency of UQCRC2 in airway epithelium of sensitized BALB/c mice prior the RWE challenge increased the Ag-induced accumulation of eosinophils, mucin levels in the airways, and bronchial hyperresponsiveness. Deficiency of UQCRC1, another oxidative damage-sensitive complex III protein, did not significantly alter cellular ROS levels or the intensity of RWE-induced airway inflammation. These observations suggest that preexisting mitochondrial dysfunction induced by oxidant environmental pollutants is responsible for the severe symptoms in allergic airway inflammation. These data also imply that mitochondrial defects could be risk factors and may be responsible for severe allergic disorders in atopic individuals.
Dendritic Cells Support the In Vivo Development and Maintenance of NK Cells via IL-15 Trans-Presentation
Authors: Castillo Eliseo F. , Stonier Spencer W. , Frasca Loredana , Schluns Kimberly S.
Published: September 2009

IL-15 is a key component that regulates the development and homeostasis of NK cells and is delivered through a mechanism termed trans-presentation. During development, multiple events must proceed to generate a functional mature population of NK cells that are vital for tumor and viral immunity. Nevertheless, how IL-15 regulates these various events and more importantly what cells provide IL-15 to NK cells to drive these events is unclear. It is known dendritic cells (DC) can activate NK cells via IL-15 trans-presentation; however, the ability of DC to use IL-15 trans-presentation to promote the development and homeostatic maintenance of NK cell has not been established. In this current study, we show that IL-15 trans-presentation solely by CD11c+ cells assists the in vivo development and maintenance of NK cells. More specifically, DC-mediated IL-15 trans-presentation drove the differentiation of NK cells, which included the up-regulation of the activating and inhibitory Ly49 receptors. Although these cells did not harbor a mature CD11bhigh phenotype, they were capable of degranulating and producing IFN-γ upon stimulation similar to wild-type NK cells. In addition, DC facilitated the survival of mature NK cells via IL-15 trans-presentation in the periphery. Thus, an additional role for NK-DC interactions has been identified whereby DC support the developmental and homeostatic niche of NK cells.
Differential Effect of IL-27 on Developing versus Committed Th17 Cells
Authors: El-behi Mohamed , Ciric Bogoljub , Yu Shuo , Zhang Guang-Xian , Fitzgerald Denise C. , Rostami Abdolmohamad
Published: September 2009

IL-27 counters the effect of TGF-β+IL-6 on naive CD4+ T cells, resulting in near complete inhibition of de novo Th17 development. In contrast, little is known about the effect of IL-27 on already differentiated Th17 cells. A better understanding of how IL-27 regulates these cells is needed to evaluate the therapeutic potential of IL-27 in Th17 cells-associated diseases. In this study, we show that IL-27 had surprisingly little effect on committed Th17 cells, despite its expression of a functional IL-27R. Contrary to de novo differentiation of Th17 cells, IL-27 did not suppress expression of retinoid-related orphan receptor (ROR)γt or RORα in committed Th17 cells. Consistent with this finding, the frequency of committed Th17 cells and their cytokine secretion remained unaffected by IL-27. Both memory Th17 cells (CD4+CD25−CD62Llow) that developed in vivo and encephalitogenic Th17 cells infiltrating the CNS of mice developing experimental autoimmune encephalomyelitis produced similar amounts of IL-17A when reactivated with IL-23 in the absence and presence of exogenous IL-27. Finally, IL-27 failed to suppress encephalitogenicity of Th17 cells in an adoptive transfer of experimental autoimmune encephalomyelitis. Analysis ex vivo of transferred Th17 cells in the spleen and CNS of recipient mice showed that cells retained similar phenotype irrespective of whether cells were treated or not with IL-27. Our data demonstrate that in contrast to inhibition of de novo differentiation of Th17 cells, IL-27 has little or no effect on committed Th17 cells. These findings indicate that therapeutic applications of IL-27 might have a limited efficacy in inflammatory conditions where aggressive Th17 responses have already developed.
DrosophilaGlycoprotein 93 Is an Ortholog of Mammalian Heat Shock Protein gp96 (grp94, HSP90b1, HSPC4) and Retains Disulfide Bond-Independent Chaperone Function for TLRs and Integrins
Authors: Morales Crystal , Wu Shuang , Yang Yi , Hao Bing , Li Zihai
Published: September 2009

Mammalian heat shock protein gp96 is an obligate chaperone for multiple integrins and TLRs, the mechanism of which is largely unknown. We have identified gp93 in Drosophila having high sequence homology to gp96. However, no functions were previously attributed to gp93. To determine whether gp93 and gp96 are functionally conserved, we have expressed gp93 in gp96-deficient mouse cells. Remarkably, the Drosophila gp93 is able to chaperone multiple murine gp96 clients including integrins α4, αL, and β2 and TLR2 and TLR9. This observation has led us to examine the structural basis of the chaperone function of gp96 by a close comparison between gp96 and gp93. We report that whereas gp96 undergoes intermolecular disulfide bond formation via Cys138, gp93 is unable to do so due to the absence of a cysteine near the same region. However, abrogation of disulfide bond formation by substituting C with A (C138A) in gp96 via site-directed mutagenesis did not compromise its chaperone function. Likewise, gp93 chaperone ability could not be improved by forcing intermolecular bond formation between gp93 N termini. We conclude that gp93 is the Drosophila ortholog of gp96 and that the chaperone function of the two molecules is conserved. Moreover, gp96 N-terminal disulfide bond formation is not critical for its function, underscoring the importance of N-terminal dimerization via non-disulfide bond-mediated interactions in client protein folding by gp96. Further study of gp96 from an evolutionary angle shall be informative to uncover the detailed mechanism of its chaperone function of client proteins in the secretory pathway.
Enhanced Dendritic Cell-Induced Immune Responses Mediated by the Novel C-Type Lectin Receptor mDCAR1
Authors: Kaden Stefan A. , Kurig Stefanie , Vasters Katrin , Hofmann Kay , Zaenker Kurt S. , Schmitz Juergen , Winkels Gregor
Published: September 2009

The dendritic cell (DC) immunoreceptors (DCIR) and DC-immunoactivating receptors (DCAR) represent a subfamily of cell surface C-type lectin receptors (CLR), whose multifunctional capacities range from classical Ag uptake and immunoregulatory mechanisms to the involvement in DC ontogeny. On the basis of the generation of specific mAbs, we functionally characterized mouse DCAR1 (mDCAR1) as a member of the DCIR/DCAR family. Expression of mDCAR1 was strongly tissue dependent. mDCAR1 expression on DCs was restricted to the CD8+ DC subset in spleen and thymus and on subpopulations of CD11b+ myeloid cells in bone marrow and spleen, whereas the molecule was not detectable on both cell types in lymph nodes and peripheral blood. With respect to the function of CLRs as pattern recognition receptors, Ag delivered via mDCAR1 was internalized, was trafficked to early and late endosomes/lysosomes and, as a consequence, induced cellular and humoral responses in vivo even in the absence of CD40 stimulation. Intriguingly, upon triggering mDCAR1, CD8+ DCs increased the secretion of bioactive IL-12, whereas IL-10 release is markedly reduced, thereby indicating that Ag recognized by mDCAR1 induces enhanced proinflammatory responses. These data indicate that mDCAR1 is a functional receptor on cells of the immune system and provides further insights into the regulation of immune responses by CLRs.
Cutting Edge: OX40 Agonists Can Drive Regulatory T Cell Expansion if the Cytokine Milieu Is Right
Authors: Ruby Carl E. , Yates Melissa A. , Hirschhorn-Cymerman Daniel , Chlebeck Peter , Wolchok Jedd D. , Houghton Alan N. , Offner Halina , Weinberg Andrew D.
Published: September 2009

We report that OX40 stimulation drives all lineages of CD4 T cell development, including regulatory T cells (Tregs), and the plasticity of the response is dependant on local cytokines. In TGF-β1-treated cultures, an OX40 agonist increased IFN-γ and IL-4 production and diverted T cells from the Treg lineage. However, cytokine blockade in the context of OX40 stimulation promoted enhanced Treg accumulation. This observation was evident in naive mice, as OX40 engagement enhanced Treg proliferation and accumulation in vivo. Lastly, OX40 agonist administration influenced experimental autoimmune encephalomyelitis disease severity in opposing directions, depending on the timing of administration. Given during Ag priming, the OX40 agonist drove Treg expansion and inhibited disease, whereas given later it enhanced T cell effector cytokine production in the CNS and exacerbated disease. Hence, OX40 signaling can augment the accumulation of all CD4 T cell lineages; however, its accentuation of immune responses may have vastly different biologic outcomes depending upon the local cytokine milieu.
Role of Endothelial TLR4 for Neutrophil Recruitment into Central Nervous System Microvessels in Systemic Inflammation
Authors: Zhou Hong , Andonegui Graciela , Wong Connie H. Y. , Kubes Paul
Published: September 2009

Brain inflammation is a frequent consequence of sepsis and septic shock. We imaged leukocyte recruitment in brain postcapillary venules induced by i.p. administration of LPS as a simple model of systemic inflammation. The i.p. injection of LPS (0.5 mg/kg) induced significant leukocyte rolling and adhesion in brain postcapillary venules of wild-type (WT) mice and more than 90% were neutrophils. However, no emigrated neutrophils were detected in brain parenchyma. High levels of TNF-α and IL-1β were detected in the plasma after LPS injection but a different profile (IL-1β but not TNF-α) was detected in the brain. LPS caused no recruitment in TLR4 knockout mice. In chimeric mice with TLR4-expressing resident cells but TLR4-deficient bone marrow-derived circulating cells, neutrophil rolling and adhesion was similar to WT mice. This observation is consistent with a requirement for resident cells in the LPS-induced neutrophil recruitment into brain microvessels. Transgenic mice engineered to express TLR4 exclusively on endothelial cells had a similar level of leukocyte recruitment in brain as WT mice in response to LPS. High dose LPS (10 mg/kg) led to neutrophil infiltration in the brain parenchyma in WT mice. High KC and MIP-2 production was observed from brain parenchyma microglial cells, and CXCR2 knockout mice failed to recruit neutrophils. However, neither neutrophil infiltration nor KC or MIP-2 was observed in endothelial TLR4 transgenic mice in response to this LPS dose. Our results demonstrate that direct endothelial activation is sufficient to mediate leukocyte rolling and adhesion in cerebral microvessels but not sufficient for emigration to brain parenchyma.
Evidence for Local Expansion of IgA Plasma Cell Precursors in Human Ileum
Authors: Yuvaraj Saravanan , Dijkstra Gerard , Burgerhof Johannes G. M. , Dammers Peter M. , Stoel Maaike , Visser Annie , Kroese Frans G. M. , Bos Nicolaas A.
Published: September 2009

IgA plays a crucial role in establishment and maintenance of mucosal homeostasis between host cells and commensal bacteria. To this end, numerous IgA plasma cells are located in the intestinal lamina propria. Whether the (immediate) precursor cells for these plasma cells can expand locally is not completely known and was studied here. The total number of IgA plasma cells in human ileal biopsies was counted. Sequence analysis of IgA VH genes from human ileal biopsies revealed the occurrence of many clonally related sequences within a biopsy, but not between different biopsies. This observation strongly argues for local expansion of IgA precursor cells. By comparing the number of unique sequences with the number of clonally related sequences within a biopsy, we estimated that ∼100–300 precursors were responsible for the 75,000 IgA-producing cells that were present per biopsy. These precursor cells must therefore have divided locally 9–10 times. Since all sequences contained mutations and most of the mutations present in clonally related sequences were shared, the IgA precursor cells must have arrived initially as mutated cells in the lamina propria. Our data show evidence for the existence of two waves of expansion for IgA-producing cells in human ileum. The first wave occurs during initial stimulation in germinal centers as evidenced by somatic hypermutations. A second wave of expansion of IgA-committed cells occurs locally within the lamina propria as evidenced by the high frequency of clonally related cells.
Lessons from Thymic Epithelial Heterogeneity: FoxN1 and Tissue-Restricted Gene Expression by Extrathymic, Endodermally Derived Epithelium
Authors: Dooley James , Erickson Matthew , Farr Andrew G.
Published: September 2009

Modeling of thymic epithelial differentiation has been guided by several important underlying assumptions. One is that within epithelial tissues derived from pharyngeal endoderm, FoxN1 expression is signature for the thymic epithelial lineage. Another is that expression of tissue-restricted Ag (TRA) is a unique feature of thymic epithelium. In this murine study, we evaluate the thymic expression of a subset of TRA, parathyroid hormone, calcitonin, and thyroglobulin, as part of an effort to better define the heterogeneity of medullary thymic epithelial cells. In this study, we demonstrate that both conventional and cystic epithelial cells display a history of FoxN1 expression using a cre-lox approach. We also document that extrathymic epithelial tissues that originate from pharyngeal endoderm also have a history of FoxN1 expression, indicating that FoxN1 expression per se is not a signature for the thymic lineage and suggesting that FoxN1 expression, whereas necessary for thymic epithelium, development, is not sufficient for this process to occur. Both cystic and conventional medullary thymic epithelial cells express these TRAs, as do extrathymic epithelial tissues that are not usually considered to be sources of these molecules. This finding supports the proposition that promiscuous gene expression is not unique to the thymus. Furthermore, the pattern of promiscuous gene expression in these extrathymic epithelia is consistent with developmental regulation processes and suggests that it is premature to discard the possibility that some promiscuous gene expression in the thymus reflects normal differentiation programs of epithelia.
Involvement of Innate and Adaptive Immunity in a Murine Model of Coronary Arteritis Mimicking Kawasaki Disease
Authors: Schulte Danica J. , Yilmaz Atilla , Shimada Kenichi , Fishbein Michael C. , Lowe Emily L. , Chen Shuang , Wong Michelle , Doherty Terence M. , Lehman Thomas , Crother Timothy R. , Sorrentino Rosalinda , Arditi Moshe
Published: September 2009

Kawasaki disease (KD) is the most common cause of acquired cardiac disease and acute vasculitis in children in the developed world. Injection of a cell wall extract isolated from Lactobacillus casei (LCCWE) into mice causes a focal coronary arteritis that histopathologically mimics the coronary lesions observed in KD patients. In this study we used this model to investigate the participation of T cells, B cells, and dendritic cells (DC) in the development of coronary arteritis. RAG1−/−, B cellnull, and wild-type (WT) mice were injected with a single dose of LCCWE (500 μg/mouse i.p.). None of the RAG1−/− mice developed coronary arteritis, whereas 70% of WT and 100% of B cellnull mice developed coronary lesions, indicating that T cells were required for lesion formation. When splenocytes isolated from LCCWE-treated mice were restimulated with LCCWE, we observed significant IFN-γ secretion in WT but not in RAG1−/− mice. Immunohistochemical staining showed F4/80+ macrophages, activated MIDC-8+ myeloid DCs (mDC), plasmacytoid DCs, and colocalization of CD3+ T cells with mDCs in coronary artery lesions, suggesting an Ag-driven process. T cells but not B cells are required for LCCWE-induced coronary arteritis. Similar to human lesions, the coronary lesions contain macrophages, activated mDCs, and plaslmacytoid DCs all in close proximity to T cells, further strengthening the relevance of this mouse model to the immunopathology of coronary disease in KD. These studies are consistent with the interpretation that macrophages and DCs may collaborate with T cells in the pathological mechanisms of coronary arteritis.
Fine Structural Variations of αβTCRs Selected by Vaccination with Natural versus Altered Self-Antigen in Melanoma Patients
Authors: Wieckowski Sébastien , Baumgaertner Petra , Corthesy Patricia , Voelter Verena , Romero Pedro , Speiser Daniel E. , Rufer Nathalie
Published: September 2009

Immunotherapy of cancer is often performed with altered “analog” peptide Ags optimized for HLA class I binding, resulting in enhanced immunogenicity, but the induced T cell responses require further evaluation. Recently, we demonstrated fine specificity differences and enhanced recognition of naturally presented Ag by T cells after vaccination with natural Melan-A/MART-1 peptide, as compared with analog peptide. In this study, we compared the TCR primary structures of 1489 HLA-A*0201/Melan-A26–35-specific CD8 T cells derived from both cohorts of patients. Although a strong preference for TRAV12-2 segment usage was present in nearly all patients, usage of particular TRAJ gene segments and CDR3α composition differed slightly after vaccination with natural vs analog peptide. Moreover, TCR β-chain repertoires were broader after natural than analog peptide vaccination. In all patients, we observed a marked conservation of the CDR3β amino acid composition with recurrent sequences centered on a glycyl-leucyl/valyl/alanyl-glycyl motif. In contrast to viral-specific TCR repertoires, such “public” motifs were primarily expressed by nondominant T cell clonotypes, which contrasted with “private” CDR3β signatures frequently found in T cell clonotypes that dominated repertoires of individual patients. Interestingly, no differences in functional avidity were observed between public and private T cell clonotypes. Collectively, our data indicate that T cell repertoires generated against natural or analog Melan-A peptide exhibited slightly distinct but otherwise overlapping and structurally conserved TCR features, suggesting that the differences in binding affinity/avidity of TCRs toward pMHC observed in the two cohorts of patients are caused by subtle structural TCR variations.
The Survival of Memory CD4+ T Cells within the Gut Lamina Propria Requires OX40 and CD30 Signals
Authors: Withers David R. , Jaensson Elin , Gaspal Fabrina , McConnell Fiona M. , Eksteen Bertus , Anderson Graham , Agace William W. , Lane Peter J. L.
Published: September 2009

Although CD4+ memory T cells reside within secondary lymphoid tissue, the major reservoir of these cells is in the lamina propria of the intestine. In this study, we demonstrate that, in the absence of signals through both OX40 and CD30, CD4+ T cells are comprehensively depleted from the lamina propria. Deficiency in either CD30 or OX40 alone reduced CD4+ T cell numbers, however, in mice deficient in both OX40 and CD30, CD4+ T cell loss was greatly exacerbated. This loss of CD4+ T cells was not due to a homing defect because CD30 × OX40-deficient OTII cells were not impaired in their ability to express CCR9 and α4β7 or traffic to the small intestine. There was also no difference in the priming of wild-type (WT) and CD30 × OX40-deficient OTII cells in the mesenteric lymph node after oral immunization. However, following oral immunization, CD30 × OX40-deficient OTII cells trafficked to the lamina propria but failed to persist compared with WT OTII cells. This was not due to reduced levels of Bcl-2 or Bcl-XL, because expression of these was comparable between WT and double knockout OTII cells. Collectively, these data demonstrate that signals through CD30 and OX40 are required for the survival of CD4+ T cells within the small intestine lamina propria.
Receptor-Mediated and Lectin-Like Activities of Carp (Cyprinus carpio) TNF-α
Authors: Forlenza Maria , Magez Stefan , Scharsack Jörn P. , Westphal Adrie , Savelkoul Huub F. J. , Wiegertjes Geert F.
Published: September 2009

Functional characterization of TNF-α in species other than mammalian vertebrates is limited, and TNF-α has been studied in a limited number of fish species, primarily in vitro using recombinant proteins. Studies on TNF-α from different fish species so far pointed to several inconsistencies, in particular with respect to some receptor-mediated activities of fish TNF-α, such as the ability to directly activate phagocytes. In the present study a comprehensive analysis of in vitro as well as in vivo biological activities of two isoforms of carp TNF-α was performed. Our results show that carp TNF-α directly primes carp phagocytes and indirectly promotes typical receptor-mediated activities such as phagocyte activation by acting via endothelial cells. Additionally, for the first time in nonmammalian vertebrate species, the lectin-like activity of fish TNF-α homologs was investigated. Our results show an evolutionary conservation of function of this receptor-independent activity of TNF-α not only in cyprinid fish, but also in perciform and salmonid fish. The role of TNF-α in vivo, during infections of carp with the blood parasite Trypanoplasma borreli, was examined using three fundamentally different but complementary approaches: (1) inhibition of TNF-α expression, (2) overexpression of TNF-α, and (3) inhibition of shedding of membrane-bound TNF-α. Our results show that, also in fish, a tight regulation of TNF-α expression is important, since depletion or excess of TNF-α can make an important difference to survival of infection. Finally, we demonstrate a crucial protective role for membrane-bound TNF-α, which has a yet unexploited function in fish.
De Novo Recruitment of Antigen-Experienced and Naive T Cells Contributes to the Long-Term Maintenance of Antiviral T Cell Populations in the Persistently Infected Central Nervous System
Authors: Zhao Jingxian , Zhao Jincun , Perlman Stanley
Published: September 2009

Mice infected with attenuated strains of mouse hepatitis virus, strain JHM, develop a chronic infection in the brain and spinal cord characterized by low levels of viral Ag persistence and retention of virus-specific CD4 and CD8 T cells at the site of infection. It is not known whether these cells are maintained by proliferation of T cells that entered the CNS during acute infection or are newly recruited from Ag-experienced or naive T cell pools. In this study, using adoptive transfer experiments and bone marrow chimeras, we show that at least some of these cells are recruited from the periphery, predominantly from the viral Ag-experienced T cell pool. Both virus-specific CD4 and CD8 T cells are functional, as assessed by cytokine expression and degranulation after peptide exposure. In addition, populations of virus-specific CD4 T cells undergo dynamic changes in the infected CNS, as previously shown for CD8 T cells, because ratios of cells responding to two CD4 T cell epitopes change by a factor of five during the course of persistence. Collectively, these results show that maintenance of T cell responses in the virus-infected CNS is a dynamic process. Further, virus-specific T cell numbers at this site of infection are maintained by recruitment from peripheral Ag-experienced and naive T cell pools.
Acrylic Elastomers
Authors: Spelta A. L.
Published: September 2009

Acrylic elastomers have the ASTM designation ACM for polymers of ethyl acrylate and other acryates. Conventionally the M indicates a polymer having a saturated chain of the polymethylene type. The repeat structure of this polymer family shows both the presence of a saturated backbone, which is responsible for the high heat and oxidation resistance, and the ester groups, which contribute to the polarity of this elastomer chain. These two main properties are not present in general‐purpose rubbers. As a result ACMs are designated as specialty elastomers and are used extensively in highly demanding applications such as automotive underhood components. However because of their wide property range, clarity and resistance to degradation by environmental forces, acrylic polymers are used in a large variety of applications.
Dyes and Dye Intermediates
Authors: Gregory Peter
Published: September 2009

The first synthetic dye, Mauveine, was discovered by Perkins in 1856, hence the dye industry is mature.The scale and growth of the dye industry depends on textiles. The most important textiles are cotton and polyester. Dyes may be classified according to their chemical structure or application method. Chemical classification is the most appropriate since it identifies dyes belonging to a group that has characteristic properties. Secondly, there are a manageable number of chemical groups (about a dozen). Also it is most used by dye chemists and technologists. Classification by use falls into the following: reactive, direct, vat, sulfur, mordant, disperse, basic, solvent, and acid dyes. Dyes are named either by their commercial trade name or by their CI (Colour Index) name. Classification by chemical name is detailed starting with the most important, azo dyes. Chemistry of the primary dye intermediates is discussed. Equipment and basics of dye manufacture are given. The toxic nature of some dyes and their intermediates is known. Dyes, because of their intense color, present special problems in effluent discharge. Removal and treatment of these wastes are detailed.
Enamels, Porcelain Or Vitreous
Authors: Faust William D.
Published: September 2009

Porcelain enamel is a glassy coating applied to various metal substrates through a high temperature fusion process, usually greater than 425°C. Enamel glass formulations are primarily alkali borosilicates. The firing, metal selection and preparation, application processes including electrostatic powder and electrophoretic deposition, and various properties are discussed. Standardized tests are listed. Porcelain enamels are used as protective, decorative,and functional coatings on steel, cast iron, aluminum, and other alloys. Products produced using porcelain enamel coatings include domestic appliances, such as ranges, refrigerators and dishwashers, and hot water tank interiors. Industrial uses include coatings for heat exchangers, chemical vessels, cast‐iron pipe, and electronic substrates. Architectural uses include interior and exterior panels. Institutional applications include chalk boards and marker boards. U.S. environmental regulations are discussed.
Insulation, Acoustic
Authors: Hirtle Parker W. , Parssinen Carol E.
Published: September 2009

Acoustic insulation may be defined s a material or construction that reduces the passage of sound into or out of a medium such as air, water or solid structure. For purposes of this article, sound is a vibratory disturbance in the air consisting of alternating compressions and rarefactions transmitted through the air waves. The term acoustic insulation covers a broad range of materials and mechanisms for the control of sound: sound absorbing materials that reduce the reflections of impinging sounds; sound blocking materials that reduce sound transmission from one location to another; vibration isolating materials and devices that reduce transmissions of vibrations from a vibrating source to potential sound‐radiating structures; and vibration dampings that reduce vibrations and sound radiation in and from materials and structures.
Insulation, Thermal
Authors: Tye Ronald P.
Published: September 2009

The applications of both open and closed cell cellular plastics for use as thermal insulation to improve energy efficiency in buildings and industrial processes are discussed. Brief details of the types and forms of cellular plastics available for such uses are included as well as typical data on thermal and other relevant performance properties and characteristics. Materials include extruded and expanded polystyrene, polyisocyanurate, polyurethane, and phenolic‐based materials. A variety of lesser used specialty materials include polyethylene, polyimide, and melamine. Temperature‐dependent properties discussed are thermal performance including aging phenomena and its required accelerated aging testing procedures, mechanical and structural performance. Flame resistance and other fire‐related issues are included as are health and safety issues. Thermal insulation is the second largest application of cellular polymers and the largest application of rigid foams. Refrigeration and residential and industrial construction are the biggest consumers of thermal insulation materials.
Elastomers, Synthetic, Survey
Authors: Morton Maurice
Published: September 2009

The purpose of this article is to provide a brief overview of the materials designated synthetic elastomers and the elastomeric or rubbery state. The term elastomer describes a material that exhibits rubbery properties, ie, that can recover most of its original dimensions after extension or compression. When elastomers are extended or stretched, the individual long‐chain molecules are partially uncoiled, but will retract when the force is removed. The properties of elastomeric materials are greatly influenced by the presence of strong interchain, ie, intermolecular forces that can result in the formation of crystalline domains. Two principal forces that govern the ability of a polymer to crystallize are the interchain attractive forces and the countervailing kinetic energy. In order to cure or vulcanize an elastomer, ie, cross‐link, certain additives are mixed and compounded with the rubber. Specific elastomers discussed are styrene‐butadiene rubber, polybutadiene, polyisoprene, elastomers resistant to age, solvent, and temperature,liquid rubber, and thermoplastics.
Safety Instrumented Systems
Authors: Summers Angela E.
Published: September 2009

One of the most common engineered safeguards in the process industry is a safety instrumented system (SIS). It uses instrumentation to detect process excursions and takes action on the process to prevent further propagation. Over the years, many terms have been used to describe types of SISs. Often, terms are used which facilitate more rapid understanding of the system's specific purpose, such as safety critical systems, high integrity protective systems, emergency shutdown systems and safety interlocks. Because they have a wide variety of functions and applications, SISs are generally designed to address specific process hazards.
Nuclear Magnetic Resonance Spectroscopy in Analysis of Chemicals Related to the Chemical Weapons Convention
Authors: Mesilaakso Markku , Meier Urs , Koskela Harri
Published: September 2009

Nuclear magnetic resonance (NMR) spectroscopy is one of the most powerful analytical techniques in organic chemistry for elucidating the molecular structures of chemicals. Moreover, an NMR spectrum may be used like a fingerprint to identify a chemical by comparing it with its reference spectrum recorded from the authentic chemical under comparable conditions. During the 20 years of international interlaboratory comparison (round‐robin) and proficiency tests for the verification of chemical disarmament, NMR spectroscopy has proved itself to be a useful complementary technique to mass spectrometry (MS) and other analytical techniques in the analysis of chemicals related to the Chemical Weapons Convention (CWC). Compared with MS, the main problems associated with NMR experiments are the lower sensitivity and the resonance overlap caused by background chemicals present in the sample. The low sensitivity of NMR experiments can be compensated in part by concentrating the NMR samples and by running long‐term experiments. The structural specificity of NMR spectra, the capability for the observation of different nuclei (
1
H,
31
P,
19
F, and
13
C), and the two‐dimensional (2D) correlation experiments make NMR spectroscopy a valuable technique to be used alongside other techniques. With regard to the chemicals listed in the Schedules of the CWC, NMR spectroscopy is useful for the analysis of almost all of them. This article describes the NMR spectroscopic methods used in the analysis of chemicals related to the CWC.
Chain of Possession of Tangible Evidence
Authors: Moenssens Andre
Published: September 2009

Laying the foundation for the admissibility of tangible physical evidence requires that, prior to its admission in evidence, the chain of possession of the evidence be established by oral or written evidence independent of the item itself. This article describes the origin and purposes of the legal principle, and describes how he appropriate foundation is to be provided to the court.
Discovery in the
U
nited
S
tates: Civil Cases
Authors: Moenssens Andre
Published: September 2009

The disclosure of expert findings to opposing parties in civil litigation is governed by codes of civil practice that are particular to each jurisdiction. For federal civil trials, the applicable provisions are set out in the Rules of Civil Procedure of 1937, as amended. The federal rules heavily favor voluntary disclosure to avoid surprise at trial. State provisions of the 50 state jurisdictions are largely modeled on the current federal statute. To avoid unwieldiness, the state rules are not discussed separately in this article.
Ethics: Codes of Conduct for Expert Witnesses
Authors: Moenssens Andre
Published: September 2009

What constitutes unethical conduct of experts is difficult to define. To attempt to do so requires an exploration of the ethical codes of the various professional societies to which the expert belongs, as well as a study of the enforcement mechanisms enacted by these organizations. This article explores the various issues that go into examining the broader principles upon which a determination is made that a person has violated the ethical principles of a society to which he belongs.
Expert Witnesses: Selection and Investigation of Credentials
Authors: Henderson Carol , Lenz Kurt W.
Published: September 2009

This article addresses the selection of an expert witness, the investigation of expert credentials, and the factors to be considered in assessing the weight an expert's opinion is likely to be given by a jury.
W

eisgram v
.

M

arley
Authors: Smith Ann C.
Published: September 2009

Weisgram v. Marley—
hereinafter
Weisgram
—is the latest of three decisions wherein the US Supreme Court explained the meaning of its
Daubert v. Merrell Dow Pharmaceuticals
ruling that, since 1993, requires experts to establish the validity of the methodology used and conclusions reached when presenting opinion evidence.
Weisgram
addressed a narrow technical issue of interest to trial and appeals courts. The case may be cited by referring to any of the following law reports: 528 U.S. 440, 120S,Ct. 1011, or 145 L.Ed.2d 958 (2000).

The US Supreme Court held that when an appeals court finds that a trial court erroneously admitted expert testimony, the tribunal is not automatically required to remand for a new trial but may, if it decides that without the offending evidence the plaintiff's case lacks sufficient evidence to support a verdict, dismiss the case outright.

This case impresses upon forensic scientists the need to carefully prepare the scientific justification that validates expert opinions reached. This article explains the Court's reasoning.
Autopsy
Authors: Saukko Pekka J. , Pollak Stefan
Published: September 2009

An autopsy, necropsy, or postmortem examination is a detailed systematic external and internal examination of a corpse usually carried out as a hospital autopsy by a pathologist or as a medicolegal one by one or more medicolegal experts to ascertain the immediate, underlying, and possible contributing causes of death, and in the latter type of an autopsy depending on the jurisdiction, also the manner of death. In some parts of the world, in particular where autopsies are rarely performed, the terms “autopsy” and “postmortem examination” are not necessarily unambiguous and may refer only to the external examination of a corpse.
Medical Malpractice
Authors: Madea Burkhard
Published: September 2009

Clear data on the epidemiology of medical malpractice are missing, at least for Germany. According to a review of the German Alliance of Patient Safety, adverse events can be expected in 5–10%, preventable adverse events in 2–4%, negligent adverse events in about 1%, and lethal outcome in about 0.1% of hospitalized patients. However, these data are not compatible with figures from civil and penal law and there seems to be a great “litigation gap”.

Data on malpractice cases are available from the files of the arbitration committees, files of liability insurance companies, and files of the institutes of forensic medicine.

The arbitration committees are, however, mainly dealing with living patients. Lethal cases are a special subgroup and the best available data source are the files of the institutes of forensic medicine. This subgroup is of special importance since death is the most severe outcome of medical malpractice and the reproach to have caused the death of a patient by medical negligence is the most severe malpractice claim.

From a retrospective multicenter study on autopsies performed in cases of medical malpractice claims (altogether 101.358 autopsies, 4450 due to medical malpractice claims), the most important data will be presented. These data are not only of a descriptive value but have utmost importance also for risk analysis and to increase patient safety. Identification and evaluation of errors as well as reporting of errors may also contribute to the prevention of errors, which is among other medical disciplines also a task of forensic medicine
Traffic Fatalities
Authors: Eriksson Anders F. , Öström Mats G.
Published: September 2009

This article describes the role of the medicolegal investigation of the fatally injured traffic victim. The main objects of this part of the crash investigation are to reveal the cause and manner of death and to reconstruct the event, but also to reveal any precrash or postcrash factors that may have contributed to the crash and/or the fatal outcome. Another purpose of the medicolegal investigation is also act as a “whistle blower” and highlight, e.g., changes in drug panorama. The article further ventilates the epidemiology of accidental/unintentional vs intentional–mainly suicidal–deaths in traffic, as well as the role of disease–causing the death and/or causing the crash. It also describes some aspects in the work to prevent traffic events resulting in death or serious bodily harm, especially the problem of drunken and drugged driving. Finally, the typical injury panorama of different categories of road users (pedestrians, motorcyclists, car drivers, and car passengers) is described in detail.