Death receptor 3 mediates necroptotic cell death

Death receptor 3 (DR3) was initially identified as a T cell co-stimulatory and pro-inflammatory molecule, but further studies revealed a more complex role of DR3 and its ligand TL1A. Although being a death receptor, DR3 gained to date predominantly attention as a contributor to inflammation-driven diseases. In our study, we investigated the cell death pathways associated with DR3. We show that in addition to apoptosis, DR3 can robustly trigger necroptotic cell death and provide evidence for TL1A-induced, DR3-mediated necrosome assembly. DR3-mediated necroptosis critically depends on receptor-interacting protein 1 (RIP1) and RIP3, the core components of the necroptotic machinery, which activate the pseudo-kinase mixed lineage kinase domain-like, the prototypic downstream effector molecule of necroptosis. Moreover, we demonstrate that DR3-mediated necroptotic cell death is accompanied by, but does not depend on generation of reactive oxygen species. In sum, we identify DR3 as a novel necroptosis-inducing death receptor and thereby lay ground for elucidating the (patho-) physiological relevance of DR3-mediated necroptotic cell death in vitro and in vivo.     

Flavonol glycosides and seed coat structure in certain species ofEpilobium — A correlation?

Summary In a survey of 239 populations ofEpilobium representing 17 taxa the following flavonol glycosides were found: myricetin 3-O-arabinoside; 3-O-glucoside; 3-O-rhamnoside; quercetin 3-O-arabinoside; 3-O-glucoside; 3-O-diglucoside; 3-O-rhamnoside; kaempferol 3-O-glucoside; and 3-O-rhamnoside. A correlation appears to exist between seed coat sculpturing as determined in a previous study using SEM techniques, and the flavonoid profiles.Acknowledgments. This study was supported in part by NSERC of Canada and The Boreal Institute for Northern Studies.     

Suppression of mitogen- and antigen-induced lymphocyte proliferation by lanthanides

Lanthanide ions (Ln3+) inhibited the proliferative response of human lymphocytes to various polyclonal mitogens and the 'purified protein derivative' (PPD) of the tuberculin antigen. Of the four Ln3+ ions tested lanthanum (La3+) was the strongest inhibitor; erbium (Er3+) and lutetium (Lu3+) were only weakly active, while samarium (Sm3+) had intermediate potency. At a concentration of 1 mM, La3+ almost completely inhibited the uptake of [3H]-thymidine by lymphocytes exposed to mitogenic agents. Trypan blue exclusion tests confirmed that the La3+ ions were not toxic. These findings may bear upon the reported anti-inflammatory properties of the lanthanides.     

Requirements for leukocyte transmigration via the transmembrane chemokine CX3CL1

The surface-expressed transmembrane CX3C chemokine ligand 1 (CX3CL1/fractalkine) induces firm adhesion of leukocytes expressing its receptor CX3CR1. After shedding by the disintegrins and metalloproteinases (ADAM) 10 and 17, CX3CL1 also acts as soluble leukocyte chemoattractant. Here, we demonstrate that transmembrane CX3CL1 expressed on both endothelial and epithelial cells induces leukocyte transmigration. To investigate the underlying mechanism, we generated CX3CR1 variants lacking the intracellular aspartate-arginine-tyrosine (DRY) motif or the intracellular C-terminus which led to a defect in intracellular calcium response and impaired ligand uptake, respectively. While both variants effectively mediated firm cell adhesion, they failed to induce transmigration and rather mediated retention of leukocytes on the CX3CL1-expressing cell layer. Targeting of ADAM10 led to increased adhesion but reduced transmigration in response to transmembrane CX3CL1, while transmigration towards soluble CX3CL1 was not affected. Thus, transmembrane CX3CL1 mediates leukocyte transmigration via the DRY motif and C-terminus of CX3CR1 and the activity of ADAM10.     

Indole-3-carbinol enhances ultraviolet B-induced apoptosis by sensitizing human melanoma cells

Indole-3-carbinol (I3C) has been found to act against several types of cancer, while ultraviolet B (UVB) is known to induce the apoptosis of human melanoma cells. Here, we investigated whether I3C can sensitize G361 human melanoma cells to UVB-induced apoptosis. We examined the effects of combined I3C and UVB (I3C/UVB) at various dosages. I3C (200 μM)/UVB (50 mJ/cm²) synergistically reduced melanoma cell viability, whereas I3C (200 μM) or UVB (50 mJ/cm²), separately, had little effect on cell viability. DNA fragmentation assays indicated that I3C/UVB induced apoptosis. Further results show that I3C/UVB activates caspase-8, −3, and Bid and causes the cleavage of poly(ADP-ribose) polymerase. Moreover, I3C decreased the expression of the anti-apoptotic protein, Bcl-2, whereas UVB increased the translocation of Bax to mitochondria. Thus, an increased Bax/Bcl-2 ratio by I3C/UVB may result in melanoma apoptosis. In conclusion, our study demonstrated that I3C sensitizes human melanoma cells by down-regulating Bcl-2. Received 5 July 2006; received after revision 25 August 2006; accepted 11 September 2006     

The flavonoid glycosides ofCornus canadensis L. and its allies in Northwestern North America

Summary The flavonoid glycoside profile ofCornus canadensis L. and its allies in Northwestern North America has been determined; quercetin 3-O-glucoside, 3-O-galactoside, 3-O-sophoroside and 3-O-gentiobioside; kaempferol 3-O-glucoside and 3-O-arabinoside. The discontinuity in distribution pattern of quercetin 3-O-gentiobioside within these taxa, associated with the phytogeography and historical factors affecting plant distribution in this area, indicates a possible polytopic and polychronistic origin of the hybrid members of the complex.Acknowledgments. This investigation was supported in part by the NRC of Canada and the Boreal Institute of Alberta for Northern Studies.     

基于共形几何代数的3D医学图像配准

随着医学图像处理技术的发展，3D／3D配准益受重视，尤其是在外科手术导航等医学应用中．学者们提出了各种3D／3D配准方法，但大多方法是采用传统代数方法进行配准，配准精度和效率都存在问题．本文利用非经典数学——共形几何代数重建了3D医学图像的位置关系约柬问题，分析了医学图像的共形几何变换，构造了新的3D医学图像配准相似测度，基于此提出了新的3D医学图像配准算法，用于CT和MR图像的3D配准．新算法中，以骨骼轮廓作为配准的基础点集，在骨骼轮廓的基础上采用共形几何代数构造共形几何体，接着采用新的3D医学图像配准相似测度进行三维数据的直接配准．实验表明新算法实现了三维数据的直接对齐，能较好地定位组织器官的三维位置．可以直观地体现配准结果．     

Calcium and magnesium deficiency and C3 fraction of complement]

The third (C3) and fourth (C4) components of complement and C3 proactivator (C3PA) were determined in 55 children with low serum levels of calcium and magnesium and 30 normal children. The concentrations of serum C3, C4 and C3PA were significantly reduced in children with double deficiences of calcium and magnesium. There were significant correlations between calcium and C3 and magnesium and C3PA. The relations between calcium, magnesium and the classical or alternate pathway of complement systems are discussed.     

Optic atrophy 3 as a protein of the mitochondrial outer membrane induces mitochondrial fragmentation

The optic atrophy 3 (OPA3) gene, which has no known homolog or biological function, is mutated in patients with hereditary optic neuropathies. Here, we identified OPA3 as an integral protein of the mitochondrial outer membrane (MOM), with a C-terminus exposed to the cytosol and an N-terminal mitochondrial targeting domain. By quantitative analysis, we demonstrated that overexpression of OPA3 significantly induced mitochondrial fragmentation, whereas OPA3 knockdown resulted in highly elongated mitochondria. Cells with mitochondria fragmented by OPA3 did not undergo spontaneous apoptotic cell death, but were significantly sensitized to staurosporine- and TRAIL-induced apoptosis. In contrast, overexpression of a familial OPA3 mutant (G93S) induced mitochondrial fragmentation and spontaneous apoptosis, suggesting that OPA3 mutations may cause optic atrophy via a gain-of-function mechanism. Together, these results indicate that OPA3, as an integral MOM protein, has a crucial role in mitochondrial fission, and provides a direct link between mitochondrial morphology and optic atrophy.     

E-cadherin and plakoglobin recruit plakophilin3 to the cell border to initiate desmosome assembly

A decrease in the levels of the desmosomal plaque protein, plakophilin3 (PKP3), leads to a decrease in desmosome size and cell-cell adhesion. To test the hypothesis that PKP3 is required for desmosome formation, the recruitment of desmosomal components to the cell surface was studied in the PKP3 knockdown clones. The PKP3 knockdown clones showed decreased cell border staining for multiple desmosomal proteins, when compared to vector controls, and did not form desmosomes in a calcium switch assay. Further analysis demonstrated that PKP3, plakoglobin (PG) and E-cadherin are present at the cell border at low concentrations of calcium. Loss of either PG or E-cadherin led to a decrease in the levels of PKP3 and other desmosomal proteins at the cell border. The results reported here are consistent with the model that PG and E-cadherin recruit PKP3 to the cell border to initiate desmosome formation.     

Serum immunoconglutinins and complement in systemicLupus erythematosus

Summary IK activity titrated by the sedimentation method in sera from patients affected with SLE was found to be negatively correlated with C4 and C3 complement factors levels. The significance of this data is discussed.Index of the abbreviations used: SLE, systemicLupus eritematosus; IK, immunoconglutinins; CH50, Total haemolytic activity; C4b, Complement factor connect with the complex AgAbCl; C3b, Complement factor connect with the complex AgAbClC4C2; C3PA, C3 proactivator in the complement activation by the alternate pathway implicated; KAF, Conglutinogen activating factor inactive the C3b in C3c and C3d. Antrypol, factor that makes C3b KAF-unreactive.     

Oncogenic protein tyrosine kinases

FLT3, a member of the class III receptor tyrosine kinases (RTKs), is preferentially expressed on the cell surface of hematopoietic progenitors, and the ligand of FLT3 (FL) is expressed as a membrane-bound or soluble form by bone marrow stroma cells. It has been disclosed that FL-FLT3 interaction plays an important role in the maintenance, proliferation and differentiation of hematopoiesis. FLT3 is also expressed in a high proportion of acute myeloid leukemia (AML) and B-lineage acute lymphoblastic leukemia cells. Activating mutations of FLT3 are the most frequent genetic lesions in AML, and AML patients with FLT3 mutations have a worse prognosis than those with normal FLT3. Exploring the mechanism by which FLT3 mutations cause autoactivation and uncontrolled signaling might lead to a better understanding of how FLT3 becomes oncogenic and provide insights for the development of new drugs.     

P2X7, NMDA and BDNF receptors converge on GSK3 phosphorylation and cooperate to promote survival in cerebellar granule neurons

Glycogen synthase kinase-3 (GSK3) is a key player in the regulation of neuronal survival. Herein, we report evidence of an interaction between P2X7 receptors with NMDA and BDNF receptors at the level of GSK3 signalling and neuroprotection. The activation of these receptors in granule neurons led to a sustained pattern of GSK3 phosphorylation that was mainly PKC-dependent. BDNF was the most potent at inducing GSK3 phosphorylation, which was also dependent on PI3K. The P2X7 agonist, BzATP, exhibited additive effects with both NMDA and BDNF to rescue granule neurons from cell death induced by PI3K inhibition. This survival effect was mediated by the PKC-dependent GSK3 pathway. In addition, ERK1/2 proteins were also involved in BDNF protective effect. These results show the function of ATP in amplifying neuroprotective actions of glutamate and neurotrophins, and support the role of GSK3 as an important convergence point for these survival promoting factors in granule neurons.     

Tyrosine 842 in the activation loop is required for full transformation by the oncogenic mutant FLT3-ITD

The type III receptor tyrosine kinase FLT3 is frequently mutated in acute myeloid leukemia. Oncogenic FLT3 mutants display constitutive activity leading to aberrant cell proliferation and survival. Phosphorylation on several critical tyrosine residues is known to be essential for FLT3 signaling. Among these tyrosine residues, Y842 is located in the so-called activation loop. The position of this tyrosine residue is well conserved in all receptor tyrosine kinases. It has been reported that phosphorylation of the activation loop tyrosine is critical for catalytic activity for some but not all receptor tyrosine kinases. The role of Y842 residue in FLT3 signaling has not yet been studied. In this report, we show that Y842 is not important for FLT3 activation or ubiquitination but plays a critical role in regulating signaling downstream of the receptor as well as controlling receptor stability. We found that mutation of Y842 in the FLT3-ITD oncogenic mutant background reduced cell viability and increased apoptosis. Furthermore, the introduction of the Y842 mutation in the FLT3-ITD background led to a dramatic reduction in in vitro colony forming capacity. Additionally, mice injected with cells expressing FLT3-ITD/Y842F displayed a significant delay in tumor formation, compared to FLT3-ITD expressing cells. Microarray analysis comparing gene expression regulated by FLT3-ITD versus FLT3-ITD/Y842F demonstrated that mutation of Y842 causes suppression of anti-apoptotic genes. Furthermore, we showed that cells expressing FLT3-ITD/Y842F display impaired activity of the RAS/ERK pathway due to reduced interaction between FLT3 and SHP2 leading to reduced SHP2 activation. Thus, we suggest that Y842 is critical for FLT3-mediated RAS/ERK signaling and cellular transformation.