bluuebunny/crossref_metadata_2025 huggingface dataset:
Hypermethylation ofFHITas a prognostic marker in nonsmall cell lung carcinoma
Authors: Maruyama Riichiroh , Sugio Kenji , Yoshino Ichiro , Maehara Yoshihiko , Gazdar Adi F.
Published: March 2004

BACKGROUND

Methylation of CpG islands in the promoter and upstream coding regions has been identified as a mechanism for transcriptional inactivation of tumor suppressor genes. The purpose of the current study was to determine the correlation between the aberrant promoter methylation of multiple genes and survival in patients with nonsmall cell lung carcinoma (NSCLC).

METHODS

The methylation status of nine genes was determined in 124 surgically resected NSCLC cases using methylation‐specific polymerase chain reaction.

RESULTS

The methylation frequencies of the genes tested in NSCLC specimens were 52% for E‐cadherin (CDH1), 41% for RAS association domain family protein (RASSF1A), 38% for fragile histidine triad (FHIT) and adenomatous polyposis coli (APC), 27% for retinoic acid receptor beta (RARβ) and H‐cadherin (CDH13), 20% forp16INK4A, 0.8% for O6‐methylguanine‐DNA‐methyltransferase (MGMT), and 0% for glutathione S‐transferase P1 (GSTP1). The survival of the patients withFHITmethylation‐positive tumors was found to be significantly shorter than that for those patients with methylation‐negative tumors (P= 0.03), even in those patients with International Union Against Cancer TNM Stage I or Stage II disease (P= 0.007). In contrast, there were no significant survival differences noted between the methylation‐positive and methylation‐negative tumors for the other genes tested. In addition, based on multivariate analyses,FHITmethylation‐positive status was found to be independently associated with poor survival (P= 0.046) and disease stage (P< 0.0001).

CONCLUSIONS

The results of the current study suggest that methylation ofFHITis a useful biomarker of biologically aggressive disease in patients with NSCLC. Cancer 2004;100:1472–7. © 2004 American Cancer Society.
A Brief Overview of CVD Research at IOPW, Technical University of Braunschweig
Authors: Arndt J. , Decker W. , Keune H. , Klippe L. , Krause U. , Metz C. , Nemetz W. , Nubian K. , Nürnberg A. , Pulver M. , Samoilenkov S.V. , Schmidt J. , Stadel O. , Stiens D. , Stolle R. , Gorbenko O.Y. , Kaul A.R. , Korsakov I.E.
Published: March 2004

In an article dedicated to Professor George Wahl on the occasion of his 65th birthday, an overview is given of CVD research at the Institute of Surface Technology and Plasma Development of Material (IOPW), and of his contributions to that research and to CVD developments over the last 35 years.
Efficient Incorporation of the Ti Component with a Novel Titanium Diolate Complex in the CVD of Ba1–xSrxTiO3 Thin Films
Authors: Lee J.S. , Hong S.H. , Woo K. , Lee W.I.
Published: March 2004

A novel titanium precursor with extended air stability and volatility is demonstrated to facilitate an efficient incorporation of the titanium component for the MOCVD of BST films. In addition to the favourable stability and volatility characteristics, the new white crystalline complex [Ti(mpd)(mdop)(μ‐OMe)]2 (Figure) has a thermal decomposition behavior comparable to those of the Ba and Sr precursors used for the deposition BST thin films from a cocktail of the three metal precursors in toluene.
MOCVD of LaMnO3 on Biaxially Textured Ni‐based Substrates in a Reducing Atmosphere
Authors: Gorbenko O.Y. , Stadel O. , Wahl G. , Kaul A.R.
Published: March 2004

The metal–organic (MO) CVD process was modified to allow the in‐situ deposition of perovskite manganites on biaxially textured Ni‐based metal tapes (RABiTS). The modification consists of the creation of the protective atmosphere by simultaneous introduction of ammonia and water. Such an atmosphere prevents the oxidation of Ni and the reduction of manganites into non‐perovskite phases. MOCVD experiments were accomplished in the band‐type reactor using the band evaporation system at 800 °C and 850 °C. Orthorhombic LaMnO3 film demonstrates (101) orientation (i.e. (001) orientation in cubic perovskite notation) minimizing lattice mismatch with the nickel substrate. The film stoichiometry, total pressure, water and ammonia flows greatly influence the quality of the film orientation. The oriented perovskite manganite films on Ni tapes can be used as buffers for high Tc superconductors for high current applications as well as magnetoresistance‐based sensors of the magnetic field.
Controlling Surface Composition and Zeta Potential of Chemical Vapor Synthesized Alumina‐Silica Nanoparticles
Authors: Sieger H. , Winterer M. , Mühlenweg H. , Michael G. , Hahn H.
Published: March 2004

Ultrafine alumina–silica powders with varying composition gradients within the particles have been produced by chemical vapor synthesis in a hot‐wall reactor using the precursors tetraethoxysilane (TEOS) and aluminiumtrisecbutylate (ASB). The surface chemistry at the particle/liquid interface in aqueous dispersions can be adjusted by varying the process parameters. The powders have been characterized by nitrogen adsorption, X‐ray diffraction (XRD), transmission electron microscopy (TEM) with local energy dispersive X‐ray (EDX) analysis, X‐ray fluorescence (XRF) analysis, and electrophoretic mobility measurements.
Growth of Praseodymium Oxide and Praseodymium Silicate Thin Films by Liquid Injection MOCVD
Authors: Aspinall H.C. , Gaskell J. , Williams P.A. , Jones A.C. , Chalker P.R. , Marshall P.A. , Smith L.M. , Critchlow G.W.
Published: March 2004

Thin films of praseodymium oxide have been deposited by liquid injection MOCVD using the volatile praseodymium alkoxide, [Pr(mmp)3] (mmp = 1‐methoxy‐2‐methyl‐2‐propanolate, OCMe2CH2OMe). The films were grown over a wide range of substrate temperatures (250–600 °C) and were found to consist predominantly of the Pr6O11 phase. Praseodymium silicate films containing ∼ 16–22 at.‐% Si were deposited over the temperature range 350–550 °C using [Pr{N(SiMe3)2}3] in the absence of any additional Si source.
Online Submission of Manuscripts (OSM)
Authors: Hitchman M.L.
Published: March 2004

It is now possible for authors to submit their manuscripts online to CVD. The system provides many advantages for authors and in this editorial Michael Hitchman explains the system and outlines its use.
Up‐regulation of muscle‐specific transcription factors during embryonic somitogenesis of zebrafish (Danio rerio) by knock‐down of myostatin‐1
Authors: Amali Aseervatham Anusha , Lin Cliff Ji‐Fan , Chen Yi‐Hsuan , Wang Wei‐Lun , Gong Hong‐Yi , Lee Chiou‐Yueh , Ko Yen‐Lin , Lu Jenn‐Khan , Her Guor Mour , Chen Thomas T. , Wu Jen‐Leih
Published: March 2004

Myostatin, a secreted growth and differentiation factor (GDF‐8) belongs to transforming growth factor (TGF‐β) superfamily that plays as a negative regulator of skeletal muscle development and growth. Recently, myostatin has been isolated from fish; however, its role in muscle development and growth remains unknown. Here, we present the expression of myostatin during development and the effects of its knock‐down on various genes such as muscle regulatory transcription factors (MRFs), muscle‐specific proteins (MSP), and insulin‐like growth factors (IGFs). The myostatin expression was found to be maternal as it starts in one‐cell stage onward. The reverse transcription‐polymerase chain reaction (RT‐PCR), in situ hybridization, and Southern and Northern blots demonstrated that the myostatin expression is not only restricted to skeletal muscle, but it expressed all the tested tissues. Expression of myostatin was effected by using antisense morpholinos resulted in significant phenotypic difference in stages 18 and 20 hours postfertilization (hpf). To confirm the specificity of myostatin morpholino, furthermore, a rescue experiment was conducted. The length as well as width of somites was increased with almost no gap in between the somites. In addition, it deserves to mention that this is a first animal model that shows changes in the size of the somites. Moreover, analyses of MRFs, MSP, and IGFs in the knock‐down embryos by RT‐PCR revealed the up‐regulation of MyoD, Myogenin, and Mck transcription, whereas IGF‐2 transcription showed mild response with no effect on IGF‐1, Desmin, and Myf5. In situ hybridization showed that there was an increase in the number of somites from 3 to 4 at 13 and 22 hpf. Taken together, these data suggest that myostatin plays a major role during myogenesis, apart from inhibition of proliferation as well as differentiation. Developmental Dynamics 229:847‐856, 2004. © 2004 Wiley‐Liss, Inc.
Reciprocal chemokine receptor and ligand expression in the human placenta: Implications for cytotrophoblast differentiation
Authors: Drake Penelope M. , Red‐Horse Kristy , Fisher Susan J.
Published: March 2004

At the onset of pregnancy, the human placenta, which forms the interface between the embryo/fetus and the mother, must rapidly develop into a life‐sustaining organ. The many unusual processes entailed in placental development include the poorly understood phenomenon of maternal tolerance of the hemiallogeneic cells of the conceptus, including, most remarkably, placental trophoblasts that invade the uterine wall. To investigate whether this fetal organ exerts control over the maternal immune system at the level of leukocyte trafficking, we examined placental expression of chemokines, well‐known cytokine regulators of leukocyte movements. In situ hybridization revealed abundant expression of 13 chemokines in the stromal but not the trophoblast compartment of chorionic villi. Potential roles for these molecules include recruitment of the resident macrophage (Hofbauer cell) population to the villi. In parallel, cytotrophoblast production of a panel of nine chemokine receptors was assessed by using RNase protection assays. The numerous receptors detected suggested the novel possibility that the paracrine actions of chemokine ligands derived from either the villous stroma or the decidua could mediate general aspects of placental development, with specific contributions to cytotrophoblast differentiation along the pathway that leads to uterine invasion. Developmental Dynamics 229:877–885, 2004. © 2004 Wiley‐Liss, Inc.
Myofibrillogenesis in the developing chicken heart: Role of actin isoforms and of the pointed end actin capping protein tropomodulin during thin filament assembly
Authors: Ehler Elisabeth , Fowler Velia M. , Perriard Jean‐Claude
Published: March 2004

Recently, important differences between myofibrillogenesis in cultured cardiomyocytes vs. the three‐dimensional setting in situ could be determined. We investigated thin filament assembly in situ by confocal microscopy of whole‐mount preparations of immunostained embryonic chicken hearts. Of interest, a distinct localisation of different actin isoforms was observed in immature thin filaments. Cardiac α‐actin is restricted to filaments with a length comparable to mature thin filaments as soon as the first contractions occur, while vascular α‐actin makes up filaments that extend toward the M‐band. The pointed‐end actin filament capping protein tropomodulin can be found initially in close association with the plasma membrane, but attains its mature localisation pattern at the ends of the thin filaments only comparatively late during myofibrillogenesis. Thus tropomodulin acts as a length stabilising element of actin filaments also in developing cardiomyocytes in situ, but plays an additional role together with membrane‐associated actin filaments in the earliest steps of myofibril assembly. Developmental Dynamics 229:745–755, 2004. © 2004 Wiley‐Liss, Inc.
Protein phosphatase 1α is required for murine lung growth and morphogenesis
Authors: Hormi‐Carver Kadija‐Kathy , Shi Wei , Liu Cathy W.Y. , Berndt Norbert
Published: March 2004

Protein phosphatase 1 (PP1) plays important roles in cell cycle control and apoptosis, two processes that impinge on morphogenesis and differentiation. Following the precedent set by other molecules regulating the cell cycle and apoptosis, we hypothesized that PP1 may have context‐specific roles in development. Therefore, we have studied the spatial and temporal expression of PP1α during murine lung development and determined the consequences of loss of PP1α function on branching morphogenesis. By using an immunohistochemical approach, we show here that PP1α was expressed throughout the epithelium and mesenchyme upon the emergence of the lung primordium on embryonic day 10, with immunostaining exclusively extranuclear. During the late pseudoglandular stage, PP1α was predominantly expressed in the distal lung epithelium, whereas the mesenchyme contained very little or no PP1α protein. Peri‐ and postnatally, PP1α immunostaining was mostly nuclear in apparently differentiated cells, as judged by colocalization with well‐known markers for lung differentiation. Exposure of fetal lung explants to antisense oligodeoxynucleotides against PP1α, resulted in decreased overall size of the cultured lung, a defect in forming new airways, lack of expression of surfactant protein C, and histologic signs of poor differentiation. These data suggest that PP1α is required for branching morphogenesis and differentiation. Developmental Dynamics 229:791–801, 2004. © 2004 Wiley‐Liss, Inc.
nlz Gene family is required for hindbrain patterning in the zebrafish
Authors: Hoyle Jacqueline , Tang Yixin P. , Wiellette Elizabeth L. , Wardle Fiona C. , Sive Hazel
Published: March 2004

This study describes the conserved nlz gene family whose members encode unusual zinc finger proteins. In the zebrafish neurectoderm, both nlz1 and the newly isolated nlz2 are expressed in the presumptive hindbrain and midbrain/hindbrain boundary, where expression of nlz1 is dependent on pax2a. In addition, nlz2 is uniquely expressed more anteriorly, in the presumptive midbrain and diencephalon. Overexpression of Nlz proteins during gastrula stages inhibits hindbrain development. In particular, ectopically expressed Nlz1 inhibits formation of future rhombomeres 2 and 3 (r2, r3), whereas neighboring r1 and r4 are not affected. Conversely, simultaneous reduction of Nlz1 and Nlz2 protein function by expression of antisense morpholino‐modified oligomers leads to expansion of future r3 and r5, with associated loss of r4. These data indicate that one function of the nlz gene family is to specify or maintain r4 identity, and to limit r3 and r5 during hindbrain formation. Developmental Dynamics 229:835–846, 2004. © 2004 Wiley‐Liss, Inc.
Knock‐in of nuclear localised β‐galactosidase reveals that the tyrosine phosphatase Ptprv is specifically expressed in cells of the bone collar
Authors: Dacquin R. , Mee P.J. , Kawaguchi J. , Olmsted‐Davis E.A. , Gallagher J.A. , Nichols J. , Lee K. , Karsenty G. , Smith A.
Published: March 2004

Ptprv is a member of the transmembrane tyrosine phosphatase gene family reported to be expressed in osteoblasts and gonads. To better define the developmental and tissue specificity of Ptprv expression, we generated knock‐in mice expressing a nuclear localised β‐galactosidase reporter under the control of resident Ptprv regulatory elements. Histochemical staining of Ptprv‐nLacZ mice revealed that Ptprv expression is readily detectable in the foetal gonadal ridge of both sexes and in adult gonads where it is localised to Sertoli cells of the testis and celomic epithelial cells of the ovaries. During early limb development, Ptprv expression is prominent in the apical ectodermal ridge of the limb bud. At latter stages of development, Ptprv is predominantly expressed in the perichondrial and periosteal region of long bones, known as the bone collar. In contrast to previous indications from in vitro studies, there is little if any expression in mature osteoblasts in vivo. Analysis of Ptprv mRNA localisation by in situ hybridization in parallel with molecular markers of chondrocytes and osteoblasts confirmed the specific expression of Ptprv in immature bone collar cells. The specificity of Ptprv expression in these cells may be a useful tool to elucidate their role in the transition of skeletal elements from cartilage template to bone. Developmental Dynamics 229:826–834, 2004. © 2004 Wiley‐Liss, Inc.
The tyrosine kinase Syk is required for light chain isotype exclusion but dispensable for the negative selection of B cells
Authors: Meade Josephine , Tybulewicz Victor L. J. , Turner Martin
Published: March 2004

In this study we set out to test whether Syk was required for negative selection of immature B cells. B cells expressing a B cell antigen receptor (BCR) transgene (3–83, anti‐H‐2Kk) underwent negative selection independently of Syk in both fetal liver organ culture and radiation chimera models. Furthermore, Syk‐independent negative selection was not reversed by transgenic overexpression of Bcl‐2. Receptor editing was not apparent in Syk‐deficient B cells, presumably as a consequence of the failure of mature edited B cells to develop in the absence of Syk. Interestingly, light chain isotype exclusion by the BCR transgene failed in the absence of Syk. We observed a dramatic reduction in the overall BCR‐mediated tyrosine phosphorylation of cellular proteins in Syk‐deficient immature B cells. However, the tyrosine phosphorylation of a number of substrates including phospholipase Cγ2, although reduced, was not completely abrogated. BCR ligation triggered an increase in calcium flux in the absence of Syk. Thus signaling events that mediate negative selection can still occur in the absence of Syk. This may be due to redundancy with zeta‐associated protein 70 (ZAP‐70), which we demonstrate to be expressed in immature B cells.
Dendritic cells from Chlamydia‐infected mice show altered Toll‐like receptor expression and play a crucial role in inhibition of allergic responses to ovalbumin
Authors: Han Xiaobing , Fan Yijun , Wang Shuhe , Yang Jie , Bilenki Laura , Qiu Hongyu , Jiao Lei , Yang Xi
Published: March 2004

Our previous study has shown that Chlamydia lung infection can inhibit local eosinophilic inflammation induced by allergen sensitization and challenge, which is correlated with altered cytokine production. In the present study, we examined the role played by dendritic cells (DC) in chlamydial infection‐mediated modulation of allergic responses. The results showed that DC freshlyisolated from Chlamydia‐infected mice (iIDC), unlike those from naive control mice (iNDC), could efficiently modulate immune responses to ovalbumin in vitro and in vivo. Co‐culture of freshly isolated DC with naive CD4 cells from T cell receptor transgenic mice (DO11.10) showed that iIDC directed Th1‐dominant, while iNDC directed Th2‐dominant, allergen‐specific CD4 T cell responses. Moreover, adoptive transfer of iIDC, but not iNDC, could inhibit systemic and local eosinophilia induced by allergen exposure. The reduction of eosinophilia was associated with a decrease in IL‐5 receptor expression on bone marrow cells and the production of IL‐5 and IL‐13 by T lymphocytes. Analysis of the DC showed that iIDC expressed significantly higher levels of mRNA for Toll‐like receptor 9 and produced more IL‐12 compared to iNDC. The data demonstrate a critical role played by DC in infection‐mediated inhibition of allergic responses.
Involvement of the CXCL12/CXCR4 pathway in the advanced liver disease that is associated with hepatitis C virus or hepatitis B virus
Authors: Wald Ori , Pappo Orit , Safadi Rifaat , Dagan‐Berger Michal , Beider Katia , Wald Hanna , Franitza Suzanna , Weiss Ido , Avniel Shani , Boaz Pal , Hanna Jacob , Zamir Gidi , Eid Ahmed , Mandelboim Ofer , Spengler Ulrich , Galun Eithan , Peled Amnon
Published: March 2004

Chronic hepatitis C virus (HCV) and hepatitis B virus (HBV) infection is accompanied by inflammation and fibrosis eventually leading to cirrhosis. The chemokine CXCL12 is involved in chronic inflammatory conditions. The role of the CXCL12/CXCR4 pathway in HCV‐ and HBV‐associated liver inflammation and fibrosis was therefore studied. The levels and tissue localization of CXCL12 in liver and plasma of HCV and HBV patients were tested using immunohistochemistry and ELISA. The expression and function of CXCR4 on liver‐infiltrating lymphocytes (LIL) were tested by FACS and transwell migration assays. We found that CXCL12 is expressed by bile duct epithelial cells in normal liver tissue. Bile duct proliferation and liver fibrosis in chronic HCV and HBV infection result in the anatomical re‐distribution of CXCL12 in the liver. Moreover, CXCL12 is up‐regulated in the endothelium of neo‐blood‐vessels formed in active inflammatory foci and is significantly elevated, compared with controls, in the plasma of patients with advanced liver fibrosis. Complementing these observations were others indicating that over 50% of LIL express CXCR4 and, in response to CXCL12, migrated and adhered to fibronectin. These observations suggest an important role for the CXCL12/CXCR4 pathway in recruitment and retention of immune cells in the liver during chronic HCV and HBV infection.
Positive and negative regulation of Fcϵ receptor I‐mediated signaling events by Lyn kinase C‐terminal tyrosine phosphorylation
Authors: Tolar Pavel , Dráberová Lubica , Tolarová Helena , Dráber Petr
Published: March 2004

Although it is known that the Src family tyrosine kinase Lyn initiates Fcϵ receptor I (FcϵRI) signaling by phosphorylation of the receptor subunits, regulation of Lyn kinase activity and its consequences for receptor signaling are incompletely understood. Using a phospho‐Lyn‐specific antiserum, we show an increased phosphorylation of the Lyn C‐terminal regulatory tyrosine and decreased Lyn kinase activity during FcϵRI‐mediated mast cell activation. Mutant Lyn, defective in the C‐terminal tyrosine, constitutively phosphorylated several substrates in resting cells, but did not cause FcϵRI internalization or spontaneous degranulation. FcϵRI‐induced signaling in the presence of constitutively active Lyn exhibited enhanced phosphorylation of the receptor subunits, Syk, LAT, Gab2, phospholipase C (PLC)γ1 and PLCγ2, and production of phosphatidylinositol 3,4,5‐trisphosphate. Although enzymatic activities of PLCγ1 and PLCγ2 were also up‐regulated, amounts of inositol 1,4,5‐trisphosphate, mobilization of intracellular calcium and degranulation were suppressed. Additionally, constitutively active Lyn was strikingly less efficient than wild‐typeLyn in restoring the receptor‐mediated calcium responses in bone marrow mast cells derived from Lyn–/– mice. These findings pinpoint the tight regulation of Lyn kinase activity as a critical event in mast cell degranulation.
Frontline: Induction of apoptosis and modulation of c‐FLIPL and p53 in immature dendritic cells infected with herpes simplex virus
Authors: Müller Dagmar B. , Raftery Martin J. , Kather Angela , Giese Thomas , Schönrich Günther
Published: March 2004

Herpes simplex virus (HSV) can perturb the function of dendritic cells (DC). The underlying mechanisms are not defined. In the present study we demonstrate that HSV induces a substantial number of immature DC to undergo apoptosis by a mechanism involving caspase‐8. We found strongly enhanced expression of TNF‐α and TRAIL but not CD95 ligand after HSV infection. Blocking experiments suggested that these classical death ligands contribute to HSV‐induced cell death of immature DC. Because uninfected DC are resistant to the apoptosis‐inducing effect of death ligands we searched fora viral "competence‐to‐die" signal. Further analysis revealed that HSV‐infected immature DC down‐regulate long cellular FLICE‐inhibitory protein (c‐FLIPL) and up‐regulate p53 whereas otherapoptosis‐regulating proteins (e.g. Bcl‐2, RIP, FADD) were not affected. Down‐regulation of c‐FLIPL was not due to diminished gene transcription or reduced mRNA stability because the level of c‐FLIPL mRNA was rather increased. Moreover, down‐regulation of c–FLIPL could not be blocked by the anti‐herpetic drug acyclovir. Finally, the underlying mechanism was also operative in human umbilical vein endothelial cells, which show a similar susceptibility to HSV infection and strength of c‐FLIPL expression. These results suggest that HSV targets c‐FLIPL protein in immature DC and other infectable cells to disrupt their function.
Increased islet antigen presentation leads to type‐1 diabetes in mice with autoimmune susceptibility
Authors: Judkowski Valeria , Krakowski Michelle , Rodriguez Enrique , Mocnick Lorraine , Santamaria Pere , Sarvetnick Nora
Published: March 2004

Granulocyte‐macrophage colony‐stimulating factor (GM‐CSF) is frequently used in preclinical and clinical protocols to modulate autoimmune responses, bone marrow transplants, and recovery from immune ablative therapies. The immunological outcome of such therapies is not fully understood. We tested the hypothesis that GM‐CSF would enhance the maturation of antigen‐presenting cells, facilitating presentation of β‐cell autoantigens to autoreactive T cells. We found that islet expression of GM‐CSF greatly enhanced disease in male mice. Islet‐derived APC but not splenic APC showed markedly enhanced capacity to stimulate in vitro proliferative responses of islet‐antigen‐specific autoreactive T cells. In vivo transfer of CD8+ and CD4+ T cells demonstrate that autoreactive T cells undergo extensive division in pancreatic lymph nodes of GM‐CSF‐transgenic mice compared with wild‐type NOD male mice. Together, the results presented here demonstrate that expression of GM‐CSF in the pancreas can enhance autoimmunity in disease‐susceptible mice.
CD4+ effector T cell distribution in vivo: TGF‐β1/TGF‐β receptor II interaction during activation mediates accumulation in the target tissue by preferential proliferation
Authors: Bode Ulrike , Tiedemann Kerstin , Westermann Jürgen
Published: March 2004

CD4+ effector T cells generated in mesenteric lymph nodes (mLN) leave into the blood. Although they enter many tissues, mLN effector T cells are later found preferentially in the gut,the drainage area of mLN. We show in the rat that this is not an intrinsic property of mLN T cells. Instead, within the mLN milieu T cells are instructed by cytokines such as transforming growth factor β1 (TGF‐β1) to up‐regulate TGF‐β receptor II (TGF‐βRII) during activation. This enables effector T cells to continue proliferation upon subsequent contact with TGF‐β1 in mLNand gut, and to accumulate in the lymph node draining area, the most likely site of pathogen invasion.
Comparative analysis of human NK cell activation induced by NKG2D and natural cytotoxicity receptors
Authors: André Pascale , Castriconi Roberta , Espéli Marion , Anfossi Nicolas , Juarez Tiffany , Hue Sophie , Conway Holli , Romagné François , Dondero Alessandra , Nanni Marina , Caillat‐Zucman Sophie , Raulet David H. , Bottino Cristina , Vivier Eric , Moretta Alessandro , Paul Pascale
Published: March 2004

NKG2D and natural cytotoxicity receptors (NCR) are essential recognition structures that mediate NK cell activation. NKG2D and NCR signaling is achieved through membrane association with signaling adaptors. The adaptors that associate with NCR — such as CD3ζ, FcRγ and KARAP/DAP12 — bear intracytoplasmic immunoreceptor tyrosine‐based activation motifs that activate Syk protein tyrosine kinases. Human NKG2D associates with the DAP10 transmembrane adaptor, which bears a YxxM motif and activates the phosphatidylinositol 3‐kinase pathway. In the mouse, a short NKG2D‐S isoform, generated by Nkg2d alternative splicing, can associate with either DAP10 or KARAP/DAP12. Here, we report that neither short human NKG2D alternative transcripts nor NKG2D association with KARAP/DAP12 was detected in activated human NK cells. Despite these results, NK cell triggering by both recombinant soluble NKG2D ligands (MICA and ULBP‐1) and anti‐NCR cross‐linking antibodies inducedsimilar CD25 expression, NK cell proliferation and cytokine production. In contrast, NKG2D triggering by anti‐NKG2D antibodies did not lead to any detectable activation signals. These data thus show that target recognition via NKG2D or NCR triggers all aspects of NK activation, and pave the way for further dissection of the signaling pathways induced by NK cell recognition of ULBP‐1 and MICA.
Systemic administration of antigen‐loaded CD40‐deficient dendritic cells mimics soluble antigen administration
Authors: Hochweller Kristin , Anderton Stephen M.
Published: March 2004

The decision to mount a T cell response to antigen (Ag) is dependent on the cellular context in which the Ag is presented. Activated dendritic cells (DC) are potent stimulators of immune responses, an ability which is linked to their high expression of several costimulatory molecules. In contrast, resting DC have been implicated in the generation of self tolerance, presumably due to their reduced costimulatory capacity. However, the precise molecular basis for the choice between Ag‐induced immunity and unresponsiveness remains unclear. We show here that CD40 plays an important rolein this decision. Systemic administration of Ag‐loaded, CD40‐deficient DC failed to induce a productive primary T cell expansion and rendered mice relatively unresponsive to subsequent immunizationwith Ag in adjuvant. Using a TCR‐transgenic T cell transfer system, we found that CD40–/– DC triggered an initial T cell activation that could not be sustained, resulting in loss of Ag‐reactive T cells and reduced cytokine production by those T cells that did persist. Furthermore, administration of CD40–/– DC that had been loaded with a central nervous system autoantigen was found to protect mice from autoimmune pathology. These data implicate the CD40:CD40L interaction as a key checkpoint in the development of T cell immunity rather than tolerance.
Involvement of NKR‐P2/NKG2D in DC‐mediated killing of tumor targets: indicative of a common, innate, target‐recognition paradigm?
Authors: Alli Rajshekhar , Savithri Balasubramanian , Das Suvendu , Varalakshmi Chavvakula , Rangaraj Nandini , Khar Ashok
Published: March 2004

DC are the most efficient antigen‐presenting cells that regulate the immune response. Here, we demonstrate the expression of NK cell receptor protein‐2 (NKR‐P2) on rat and mouse DC, and we show that NKR‐P2 gets reorganized upon antigen contact. DC activated with anti‐NKR‐P2 mAb exhibit enhanced apoptotic killing of tumor targets, whereas blocking the interaction between NKR‐P2 and its ligand with rNKR‐P2 abrogated apoptotic killing, suggesting NKR‐P2 to function as an activating molecule on DC. In vivo injection of anti‐NKR‐P2 mAb augmented DC activity and delayed tumor progression. NKR‐P2 signaling involved Ca2+ influx, culminating in the expression of the apoptosis‐inducing molecule, TNF‐α. Taken together, these observations suggest that NKR‐P2 (the rat orthologue of human NKG2D) acts as a target‐recognition molecule on DC.
Frontline: Optimal T cell activation requires the engagement of CD6 and CD166
Authors: Hassan Namir J. , Barclay A. Neil , Brown Marion H.
Published: March 2004

The T cell surface glycoprotein, CD6 binds CD166 in the first example of an interaction between a scavenger receptor cysteine‐rich domain and an immunoglobulin‐like domain. We report that in human these proteins interact with a KD =0.4–1.0 μM and Koff ≥0.4–0.63 s–1, typical of many leukocyte membrane protein interactions. CD166 also interacts in a homophilic manner but with around 100‐fold lower affinity (KD =29–48 μM and Koff ≥ 5.3 s–1). At concentrations, that will block the CD6/CD166 interaction, soluble monomeric CD6 and CD166 inhibit antigen‐specific human T cell responses. This is consistent with extracellular engagement between CD6 and CD166 being required for an optimal immune response.
Complement activation by both classical and alternative pathways is critical for the effector phase of arthritis
Authors: Hietala Max Albert , Nandakumar Kutty S , Persson Linda , Fahlén Susann , Holmdahl Rikard , Pekna Marcela
Published: March 2004

To analyze the role of the classical and alternative pathways of complement activation in the effector phase of arthritis, we have induced arthritis in C3‐ and factor B (FB)‐deficient (C3–/– and FB–/–) DBA/1J mice using well‐defined monoclonal IgG2b and IgG2a antibodies to type II collagen. In control DBA/1J mice, severe swelling of the joints, destruction of cartilage and erosion of bone developed very rapidly with a 100% incidence and a peak on days 7–10. Although 75% of C3–/– mice developed arthritis, the clinical severity was very mild and the onset was delayed. Severity of arthritis in FB–/– mice ranked intermediate in comparison with C3–/– and control mice with an incidence of 100%. Immunohistochemical analysis of the inflamed joints demonstrated substantial reduction in macrophage and neutrophilic leukocyte infiltration in both C3–/– and FB–/– mice, thereby confirming the clinical findings. We conclude that both the classical and the alternative pathways of complement activation are involved in the effector phase of arthritis.
Maintenance of memory CTL responses by T helper cells and CD40‐CD40 ligand: antibodies provide the key
Authors: Bachmann M. F. , Hunziker Lukas , Zinkernagel R. M. , Storni T. , Kopf M.
Published: March 2004

Vol. 34(2) 2004, pp 317–326, DOI 10.1002/eji.200324717

The authors would like to add the following information regarding the viral doses used in the experiments:

All experiments with recombinant vaccinia virus were performed with a standard dose of 2×106 pfu. All experiments with LCMV WE were performed with a standard dose of 200 pfu. The intravenous route of infection was used for both viruses.

See original article http://dx.doi.org/10.1002/eji.200324717
Hydrothermal Syntheses, Crystal Structures, Magnetism and Fluorescence Quenching of Oxamidato‐Bridged Pentanuclear CuII4LnIII Complexes Containing Macrocyclic Ligands (Ln = Eu, Tb) and the Crystal Structure of a Hexanuclear NiII5SmIII Complex
Authors: Sun Ya‐Qiu , Liang Mao , Dong Wen , Yang Guang‐Ming , Liao Dai‐Zheng , Jiang Zong‐Hui , Yan Shi‐Ping , Cheng Peng
Published: March 2004

Three novel oxamidato‐bridged complexes incorporating a macrocyclic oxamide of formula [(CuL)3{CuL(C2H5OH)}Eu(H2O)](ClO4)3·1.5H2O (1), [(CuL)3{CuL(C2H5OH)}Tb(H2O)](ClO4)3·2H2O (2) and [(NiL)5Sm](ClO4)3·2H2O (3) (CuL and NiL, H2L = 2,3‐dioxo‐5,6,14,15‐dibenzo‐1,4,8,12‐tetraazacyclopentadeca‐7,13‐dien), have been hydrothermally synthesized and structurally characterized. In these complexes, the central LnIII and external copper (or nickel) ions are bridged by macrocyclic oxamide groups. The EuIII (or TbIII) ion of 1 (or 2) resides in a distorted tricapped trigonal prismatic environment surrounded by eight oxygen atoms from four oxamide groups and one oxygen atom from a water molecule, whereas the SmIII ion of 3 resides in a distorted bicapped square antiprismatic environment surrounded by ten oxygen atoms from five oxamide groups. Furthermore, there are hydrogen bonding interactions in 1 and 2 between perchlorate, ethanol and water moieties. Especially for 1 and 2, O−H···O and weak coordination of ClO4− groups link the pentanuclear fragments and perchlorate ions to form a 1‐D supramolecular architecture. The fluorescence of EuIII and TbIII are almost completely quenched in 1 and 2. The magnetic properties of 1 and 2 have been characterized. (© Wiley‐VCH Verlag GmbH & Co. KGaA, 69451 Weinheim, Germany, 2003)
Macrocyclic and Open‐Chain CuII‐4f (4f = GdIII, CeIII) Complexes with Planar Diamino Chains: Structures and Magnetic Properties
Authors: Costes Jean‐Pierre , Dahan Françoise , Novitchi Ghenadie , Arion Vladimir , Shova Sergiu , Lipkowski Janusz
Published: March 2004

Until now, the known structures of heterodinuclear Cu−Ln complexes involve compartmental Schiff‐base ligands with saturated diamino chains. We report here two Cu−Ln complexes (Ln = Gd or Ce) which are the first structurally characterized Cu−Ln entities involving S‐methylisothiosemicarbazide as the diamino bridge, with open chain and macrocyclic ligands. The macrocyclic Cu−Gd entity is a well isolated dinuclear complex whereas two hydrogen bonds join together a pair of binuclear Cu−Ce complexes to give tetranuclear {Cu−Ce···Ce−Cu} entities. The Cu−Gd complexes [H = ‐J(SCuSGd)] exhibit ferromagnetic interactions between 5 and 5.5 cm−1. Since the Cu−Ce complexes do not follow the Curie law due to the presence of orbital degeneracy, an analysis based on a simple spin‐only Hamiltonian is not possible. Nevertheless, the EPR spectra of the Cu−Ce complexes do confirm the existence of magnetic interactions. (© Wiley‐VCH Verlag GmbH & Co. KGaA, 69451 Weinheim, Germany, 2004)
Hydrolytic Cleavage of DNA by Ternary Amino Acid Schiff Base Copper(II) Complexes Having Planar Heterocyclic Ligands
Authors: Reddy Pattubala A. N. , Nethaji Munirathinam , Chakravarty Akhil R.
Published: March 2004

The ternary copper(II) complexes [Cu(salgly)L] (L = phen, 1; dpq, 2), [Cu(salala)L] (L = phen, 3; dpq, 4) and [Cu(salphe)L] (L = phen, 5; dpq, 6), where salgly, salala and salphe are tridentate Schiff‐base ligands derived from the condensation of salicylaldehyde with glycine, L‐alanine and L‐phenylalanine, respectively, are prepared and their nuclease activity studied (phen, 1,10‐phenanthroline; dpq, dipyridoquinoxaline). The crystal structure of 3 displays a distorted square‐pyramidal (4+1) coordination geometry in which the ONO‐donor Schiff base is bonded to the metal atom in the basal plane. The chelating phen ligand displays an axial−equatorial mode of bonding. The complexes exhibit a d−d band near 670 nm and a charge transfer band near 370 nm in methanol. The one‐electron paramagnetic complexes display axial EPR spectra in DMF glass at 77 K, indicating a {d−}1 ground state. The complexes are redox‐active and exhibit a quasi‐reversible CuII/CuI couple in DMF at approximately −0.6 V vs. SCE. They show catalytic activity in the oxidation of ascorbic acid by molecular oxygen. The ability of the complexes to bind calf thymus (CT) DNA follows the order: 2 ≈︁ 4 ≈︁ 6 > 1 ≈︁ 3 ≈︁ 5. Complexes 1−6 show oxidative DNA cleavage activity in the presence of mercaptopropionic acid as a reducing agent. All the complexes show hydrolytic cleavage activity in the absence of light or any reducing agent. The oxidative and hydrolytic DNA cleavage efficiencies follow the order: 2 ≈︁ 4 ≈︁ 6 > 1 ≈︁ 3 ≈︁ 5. The dpq complexes, which have a greater DNA binding ability, display enhanced nuclease activity than their phen analogues. The hydrolytic DNA cleavage rate of 1.8 h−1, observed for 2, is significantly high relative to most of the known copper‐based synthetic hydrolases. Mechanistic pathways involved in the nuclease activity of the complexes are discussed. (© Wiley‐VCH Verlag GmbH & Co. KGaA, 69451 Weinheim, Germany, 2004)
Rhodium(I), Palladium(II) and Platinum(II) Coordination Chemistry of the Short‐Bite Chiral Ligands (Sc)‐N,N‐Bis(diphenylphosphanyl)‐sec‐butylamine, (Ra,Ra)‐N,N‐Bis(binaphthylphosphonito)phenylamine and (Ra,Sc)‐N‐(Diphenylphosphanyl)‐N‐(binaphthylphosphonito)‐sec‐butylamine
Authors: Calabrò Gianpiero , Drommi Dario , Graiff Claudia , Faraone Felice , Tiripicchio Antonio
Published: March 2004

The new short‐bite chiral ligands (Sc)‐N,N‐bis(diphenylphosphanyl)‐sec‐butylamine [(Sc)‐1], (Ra,Ra)‐N,N‐bis(binaphthylphosphonito)phenylamine [(Ra,Ra)‐2] and (Ra,Sc)‐N‐(diphenylphosphanyl)‐N‐(binaphthylphosphonito)‐sec‐butylamine [(Ra,Sc)‐3] have been treated with rhodium(I), palladium(II) and platinum(II) substrates. The (Sc)‐1 ligand reacts with [Pd(C6H5CN)2Cl2], [Pt(COD)Cl2] and [Rh(COD)(THF)2]PF6 to afford the products [Pd(Sc‐1)Cl2] (4), [Pt(Sc‐1)Cl2] (5) and [Rh(COD)(Sc‐1)]PF6 (6), respectively, in high yields. By bubbling CO to a CH2Cl2 solution of 6, we formed the dicarbonyl species [Rh(CO)2(Sc‐1)]PF6 (7), which is stable only in solution under CO. Compounds 4 and 5 were characterized additionally by X‐ray diffraction studies. The ligand (Ra,Ra)‐2 reacts with [Pd(C6H5CN)2Cl2] and [Pt(COD)I2] to give [Pd(Ra,Ra‐2)Cl2] (8) and [Pt(Ra,Ra‐2)I2] (9), respectively; the reaction with [Rh(CO)2Cl]2 or [Rh(COD)(THF)2]PF6 affords a mixture of mono‐ and binuclear compounds, in which the ligand is oxidized to the corresponding phosphonate or is partially hydrolysed. The compounds [Pd(Ra,Sc‐3)Cl2] (10) and [Pt(Ra,Sc‐3)I2] (11) were obtained in the analogous reactions using the asymmetric (Ra,Sc)‐3 ligand. This ligand reacted with [Rh(CO)2Cl]2 in a 2:1 molar ratio in hexane to give a mixture of products in almost equal amounts. The 31P{1H} NMR spectrum recorded in CDCl3 allowed us to identify the chelate [Rh(Ra,Sc‐3)(CO)Cl] (12); the nature of the accompanying dinuclear species 13 is not clear. The catalytic systems formed by [Rh(acac)(CO)2] and the ligands (Ra,Ra)‐2 and (Ra,Sc)‐3, at ligand/metal ratios of 1:1 and 1:2, were unstable under the experimental conditions used (CO/H2 pressure of 50 atm and temperature of 40 °C) for the hydroformylation of styrene; instead, metallic rhodium was afforded together with unidentified decomposition products. (© Wiley‐VCH Verlag GmbH & Co. KGaA, 69451 Weinheim, Germany, 2004)
The Synthesis of Stable Intramolecularly Dialkylamino‐Coordinated Silenes and Their Methyl Iodide Induced Isomerization into Cyclic Aminosilanes
Authors: Mickoleit Martin , Schmohl Kathleen , Michalik Manfred , Oehme Hartmut
Published: March 2004

The reaction of the (dichloromethyl)oligosilanes R(Me3Si)2SiCHCl2 4a−d (4a: R = Me; 4b: R = tBu; 4c: R = Ph; 4d: R = SiMe3) with 8‐(dimethylamino)‐1‐naphthyllithium leads to the intramolecularly donor‐stabilized silenes 1a−d. Whereas 1a,b, and d were isolated as thermally stable yellow crystalline compounds, 1c could not be separated in a pure form but was characterized by NMR spectroscopy and mass spectroscopy as well as by its chemical reactivity with a variety of substrates. Dynamic 1H NMR spectroscopic studies of 1a and 1d revealed coalescence of the signals of the two CSiMe3 groups. In agreement with theoretical calculations, this effect was interpreted as the result of internal rotations about the silene double bond. These findings and the known relatively long Si=C distances for 1d, 2 and 3 help establish the ylide‐like nature of the silenes prepared. Treatment of 1a−d with water produced the silanols 9a−d. Treatment of 1a,b, and d with methyl iodide caused a rearrangement of the silenes, one methyl substituent formally migrating from the coordinated dimethylamino group to the silene carbon atom producing the respective cyclic aminosilanes 11a,b, and d. As the outcome of the reaction of 1d with benzaldehyde, 2,2‐bis(trimethylsilyl)vinylbenzene (14) and a mixture of the two isomers of the cyclodisiloxane 13 were obtained. (© Wiley‐VCH Verlag GmbH & Co. KGaA, 69451 Weinheim, Germany, 2004)
Tetranuclear (Borole)rhodium Clusters with Hydrido and Methylthiolato Ligands − Structures of [{Rh(C4H4BMe)}4(µ3‐H)(µ3‐I)3] and of [{Rh(C4H4BPh)}2(µ‐SMe)(µ3‐SMe)]2 (2 Rh−Rh)
Authors: Herberich Gerhard E. , Eckenrath Hartmut J. , Wagner Trixie , Wang Ruimin
Published: March 2004

The heterocubanes [Rh(C4H4BR)(µ3‐I)]4 (1a: R = Ph; 1b: R = Me) can be hydrogenated in THF (suspension, 1 bar H2, 20 °C) to give the µ3‐hydrido clusters [{Rh(C4H4BR)}4(µ3‐H)(µ3‐I)3] (2a,b). The structure of 2b consists of a Rh4(µ3‐H)(µ3‐I)3 core with idealized C3v symmetry and four capping η5‐borole ligands. The Rh−Rh distances within the Rh3(µ3‐H) cluster unit amount to 2.9448(6), 2.9647(6), and 2.9724(6) Å; the µ3‐H ligand is 0.064(5) Å above the Rh3 plane with a mean Rh−H bond length of 1.83(5) Å. The tris(acetonitrile) salts [Rh(C4H4BR)(NCMe)3]BF4 (4a,b) react with NaSMe to give the methylthiolato complexes [Rh(C4H4BR)(SMe)]x (3a,b). In the crystal 3a possesses a centrosymmetric stepped‐ladder structure [{Rh(C4H4BPh)}2(µ‐SMe)(µ3‐SMe)]2 (2Rh−Rh) (3a‐Rh4), with a Rh−Rh bond length of 2.9233(11) Å. In solution a dissociation equilibrium is observed with a predominating dinuclear complex [Rh(C4H4BPh)(µ‐SMe)]2 (3a‐Rh2); in pyridine a closely related solvate [Rh(py)(C4H4BPh)(µ‐SMe)]2 (5) with a static Rh2(μ‐SMe)2 core is formed. (© Wiley‐VCH Verlag GmbH & Co. KGaA, 69451 Weinheim, Germany, 2004)
The First Silver(I) Side‐On η2‐Coordination Network Formed by Self‐Assembly of Bis(pyrrol‐2‐ylmethylene)hydrazine
Authors: Yang Lanying , Shan Xufeng , Chen Qingqi , Wang Zhengping , Ma Jin Shi
Published: March 2004

This paper reports the first silver(I) network polymer formed by the self‐assembly of bis(pyrrol‐2‐ylmethylene)hydrazine with silver nitrate, in which the silver centers are bonded to the C−N bonds of the pyrrole rings in an η2‐fashion. An X‐ray crystal structure analysis revealed that the self‐assembly of the bis(pyrrol‐2‐ylmethylene)hydrazine ligand with AgI gives a 2D sheet, which is further assembled to form a 3D network through weak intermolecular hydrogen bonding between the protons of the pyrrole rings and the oxygen atoms of the nitrate ions. (© Wiley‐VCH Verlag GmbH & Co. KGaA, 69451 Weinheim, Germany, 2004)
Synthesis, Characterization and Reactivity of New (μ‐Aminocarbyne)diruthenium Complexes Containing Alkynylimino Ligands
Authors: Busetto Luigi , Marchetti Fabio , Zacchini Stefano , Zanotti Valerio
Published: March 2004

Addition of acetylides R′C≡CLi [R′ = Tol, Ph, Me3Si, nBu, C(Me)=CH2] to the diruthenium complexes [Ru2{μ‐CN(Me)(R)}(μ‐CO)(CO)(NCCMe3)(Cp)2]+ (R = Xy, 2a; Bz, 2b; Me, 2c) results in the formation of [Ru2{μ‐CN(Me)(R)}(μ‐CO)(CO){N(H)=C(CMe3)(C≡CR′)}(Cp)2]+ (R = Xy, R′ = Tol, 3a; R = Bz, R′ = Tol, 3b; R = Me, R′ = Tol, 3c; R = Xy, R′ = Ph, 4a; R = Bz, R′ = Ph, 4b; R = Me, R′ = Ph, 4c; R = Bz R′ = H, 5; R = Xy, R′ = nBu, 6; R = Xy, R′ = [C(Me)=CH2], 7), which contain an alkynylimino ligand, formed from addition of the acetylide to the Me3CCN ligand. Structural and spectroscopic studies show that all these complexes contain the two Cp ligands in a relative cis arrangement, whereas two isomers arise from the different orientation of the substituents on the bridging aminocarbyne μ‐CN(Me)(R) when R ≠ Me. Addition of secondary amines to 3−7 results in a stereo‐ and regioselective cis amination of the C−C triple bond to give the new complexes [Ru2{μ‐CN(Me)(R)}(μ‐CO)(CO){N(H)=C(CMe3)C(H)=C(NMe2)(Tol)}(Cp)2]+ (R = Xy, 8a; Bz, 8b; Me, 8c), [Ru2{μ‐CN(Me)(Xy)}(μ‐CO)(CO){N(H)=C(CMe3)C(H)=C(NC5H10)(Tol)}(Cp)2]+(9) and [Ru2{μ‐CN(Me)(Xy)}(μCO)(CO){N(H)=C(CMe3)C(H)=C(NMe2)C(Me)=CH2)}(Cp)2]+ (10) containing the hitherto unknown new imino‐2‐en‐3‐amine ligand. The solid‐state structures of these products have been analyzed by X‐ray analysis and, in solution, by NOE spectroscopic studies. (© Wiley‐VCH Verlag GmbH & Co. KGaA, 69451 Weinheim, Germany, 2004)
Cloning and analysis of axolotl ISL2 and LHX2 LIM‐homeodomain transcription factors
Authors: Showalter Aaron D. , Yaden Benjamin C. , Chernoff Ellen A.G. , Rhodes Simon J.
Published: March 2004

Summary: We cloned and characterized the ISL2 and LHX2 LIM‐homeodomain transcription factors of the Mexican salamander, or axolotl, Ambystoma mexicanum. Using a degenerate PCR approach, partial cDNAs representing five LIM‐homeodomain genes were cloned, indicating conservation of this class of transcription factors in urodeles. Full‐length cDNAs for Isl2 and Lhx2 were identified and sequenced. The predicted ISL2 and LHX2 proteins are well conserved, especially in the LIM and DNA‐binding domains. The Isl2 and Lhx2 genes are expressed at all examined stages of embryogenesis and display tissue‐restricted expression patterns in adults. In functional tests, axolotl LHX2 was inactive compared to homologous mammalian factors and adopted unusual DNA/protein complexes. However, axolotl ISL2 bound and induced transcription from the rat insulin gene. These experiments demonstrate conservation of key developmental regulatory proteins in salamanders and will allow future studies of their potential roles in the molecular regulation of tissue regeneration in such species. genesis 38:110–121, 2004. © 2004 Wiley‐Liss, Inc.