Structure and evolutionary origin of the human granzyme H gene

Among the molecules proposed to be involved In cytotoxic T lymphocyte (CTL), natural killer (NK) and lymphokine activated killer (LAK) cell-mediated lysis are the granzymes, a family of serine proteases stored in the cytoplasmic granules of CTLs, NK and LAK cells. In addition to the granzymes A and B, a third member of this family has been cloned in man and designated granzyme H. We present the complete gene sequence including the 5' promoter region and demonstrate that the granzyme H sequence represents a functional gene expressed In activated T cells. Granzyme H shows the highest degree (>54%) of amlno acid sequence homology with granzyme B and cathepsin G and, like these genes, consists of five exons separated by introns at equivalent positions. The evolutionary history of granzyme H has been analyzed by reconstructing an evolutionary tree for granzyme sequences. We provide evidence that Interlocus recombination between the ancestral genes of granzyme B and granzyme H occurred about 21 million years ago, leading to a replacement of exon 3, Intron 3 and part of exon 4 in human granzyme H by human granzyme B sequences. Our results suggest that the ancestral gene of granzyme H is more closely related to cathepsin G and granzyme B than to the murine granzymes C to G; Thus, granzyme H does not represent a human counterpart of the known murine granzymes A to G. It diverged from cathepsin G before mammalian radiation and should, therefore, exist in other mammalian lineages as wel

Identification of Novel Functions for Hepatitis C Virus Envelope Glycoprotein E1 in Virus Entry and Assembly

International audienceHepatitis C virus (HCV) envelope glycoprotein complex is composed of E1 and E2 subunits. E2 is the receptor-binding protein as well as the major target of neutralizing antibodies, whereas the functions of E1 remain poorly defined. Here, we took advantage of the recently published structure of the N-terminal region of the E1 ectodomain to interrogate the functions of this glycoprotein by mutating residues within this 79-amino-acid region in the context of an infectious clone. The phenotypes of the mutants were characterized to determine the effects of the mutations on virus entry, replication, and assembly. Furthermore, biochemical approaches were also used to characterize the folding and assembly of E1E2 heterodimers. Thirteen out of 19 mutations led to viral attenuation or inactivation. Interestingly, two attenuated mutants, T213A and I262A, were less dependent on claudin-1 for cellular entry in Huh-7 cells. Instead, these viruses relied on claudin-6, indicating a shift in receptor dependence for these two mutants in the target cell line. An unexpected phenotype was also observed for mutant D263A which was no longer infectious but still showed a good level of core protein secretion. Furthermore, genomic RNA was absent from these noninfectious viral particles, indicating that the D263A mutation leads to the assembly and release of viral particles devoid of genomic RNA. Finally, a change in subcellular colocalization between HCV RNA and E1 was observed for the D263A mutant. This unique observation highlights for the first time cross talk between HCV glycoprotein E1 and the genomic RNA during HCV morphogenesis

ICAM-1 PROMOTES THE ABNORMAL ENDOTHELIAL CELLPHENOTYPE IN CHRONIC THROMBOEMBOLIC PULMONARYHYPERTENSION

International audienceBACKGROUND - Pulmonary endothelial cells play a key role in the pathogenesis of ChronicThromboembolic Pulmonary Hypertension (CTEPH). Increased synthesis and/or release ofIntercellular Adhesion Molecule 1 (ICAM-1) by pulmonary endothelial cells of patients withCTEPH has been recently reported, suggesting a potential role for ICAM-1 in CTEPH.METHODS - We studied pulmonary endarterectomy specimens from 172 patients with CTEPHand pulmonary artery specimens from 97 controls undergoing lobectomy for low-stage cancerwithout metastasis.RESULTS - ICAM-1 was overexpressed in vitro in isolated and cultured endothelial cells fromendarterectomy specimens. Endothelial cell (EC) growth and apoptosis resistance weresignificantly higher in CTEPH specimens than in controls (P<0.001). Both abnormalities wereabolished by pharmacological inhibition of ICAM-1 synthesis or activity. Overexpression ofICAM-1 contributed to the acquisition and maintenance of abnormal EC growth and apoptosisresistance via phosphorylation of SRC, p38 and ERK1/2 and overproduction of Survivin.Regarding the ICAM-1 E469K polymorphism, the KE heterozygote genotype was significantlymore frequent in CTEPH than in controls, but was not associated with disease severity amongpatients with CTEPH.CONCLUSIONS - ICAM-1 contributes to maintaining the abnormal endothelial cell phenotypein CTEPH

Alterations in Homologous Recombination-Related Genes and Distinct Platinum Response in Metastatic Triple-Negative Breast Cancers: A Subgroup Analysis of the ProfiLER-01 Trial

International audienc

Corrélation phénotype-génotype chez 14 patients porteurs d'une délétion de la région chomosomique 1q24q25

PARIS-BIUP (751062107) / SudocSudocFranceF

Exploring the field of NGO cooperation in Lebanon: Snapshot from the field

Non-governmental organizations (NGOs) in Lebanon face survival challenges that require them to cooperate. With many attempts falling short of their goals, this paper studied dysfunctions in NGO cooperation. An intervention-research methodology was adopted to accompany fifteen active organizations in framing cooperation dysfunctions and developing solutions. We identified major interorganizational dysfunctions grouped under strategy, decision-making, procedures, and language axes. The researchers then accompanied organizations in collective solution provision to improve their cooperation. Theoretical implications point towards a moderated relationship between intra and interorganizational improvements, and that interorganizational dysfunctions transfer into interorganizational cooperation level.Les organisations non gouvernementales (ONG) du Liban sont confrontées à des défis de survie qui les obligent à coopérer. Cet article explore les dysfonctionnements qui ont empêché plusieurs tentatives de coopération entre ONG d’atteindre leurs objectifs. Une méthode de recherche-intervention a été adoptée pour accompagner quinze organisations dans l’indentification des dysfonctionnements de coopération et l’élaboration de solutions. Nous avons identifié des dysfonctionnements inter-organisationnels majeurs regroupés sous les axes stratégie, prise de décision, procédures et langage. Les chercheurs ont ensuite accompagné les organisations dans la conception collective de solutions pour améliorer leur coopération. Les résultats pointent une relation modérée entre les améliorations intra et inter-organisationnelles, et un transfert direct des dysfonctionnements intra-organisationnels omniprésents vers le niveau de la coopération inter-organisationnelle.Las organizaciones no gubernamentales (ONG) del Líbano se enfrentan a retos de supervivencia que les obligan a cooperar. Este artículo explora los disfuncionamientos que han impedido que varios intentos de cooperación entre ONG alcancen sus objetivos. Se adoptó un método de investigación-intervención para ayudar a quince organizaciones a identificar disfuncionamientos en la cooperación y desarrollar soluciones. Se identificaron los principales disfuncionamientos interorganizacionales agrupados bajo los epígrafes de estrategia, toma de decisiones, procedimientos y lenguaje. A continuación, los investigadores ayudaron a las organizaciones a idear colectivamente soluciones para mejorar su cooperación. Los resultados apuntan a una relación moderada entre las mejoras intra e interorganizacionales, y a una transferencia directa de los disfuncionamientos interorganizacionales dominantes al nivel de cooperación interorganizacional

Straight: stochastic geometry and user history based mobility estimation

International audience5G is envisioned to support scalable networks and improved user experience with virtually zero latency and ultra broad-band service. Supporting unlimited seamless mobility is one of the key issues and also for network resource utilization efficiency. In this paper, we focus on mobility management and user equipment (UE) speed class estimation, also known as mobility state estimation (MSE). We propose a method for estimating the UE mobility which is compliant with UE history information specifications by 3GPP (3rd Generation Partnership Project). We also exploit the impact of the environment on the UE trajectory and speed when determining UE mobility state. We evaluate the effectiveness of our algorithm using realistic mobility traces and network topology of the city of Cologne in Germany provided by the Kolntrace project [1]. Results show that the speed classification of UEs can be achieved with much higher accuracy compared to existing legacy 3GPP LTE MSE procedures

Two new biologically active triterpenoidal saponins acylated with salicylic acid from Albizia adianthifolia

International audienceTwo new oleanane-type triterpene saponins, adianthifoliosides A (1) and B (2), were isolated from a 95% ethanolic extract of roots of Albizia adianthifolia. Their structures were elucidated mainly by using a combination of 600 MHz 1D and 2D NMR techniques (COSY, NOESY, TOCSY, HSQC, and HMBC) and by FABMS and HRESIMS. Compounds 1 and 2 were characterized as glycosides of acacic acid acylated by an o-hydroxybenzoyl unit. The crude saponin mixture (CSM), compounds 1 and 2 together with 3 and 4 (prosapogenins obtained from the mild alkaline hydrolysate of the CSM), were evaluated for immunomodulatory activity on the Jurkat T cell line and for hemolytic property against sheep erythrocytes. Compound 2 and, to a lesser extent, 1 and 3 were found to exhibit a dose-dependent immunomodulatory effect in the concentration range 10-2-10 µM, whereas 4 showed a lymphoproliferative activity in the same concentration range. Among the compounds tested, only 1 and 2 were found to be hemolytic. The genus Albizia comprises about 150 species widely distributed in the tropics, with the greatest diversity in Africa and Central and South America. 1 Albizia species have been reported to contain alkaloids, steroids, triter-penoid saponins, and flavonols. 2 Some saponins such as julibrosides J 1 , J 2 , and J 9 from Albizia julibrissin 3,4 possess various biological effects, such as inhibitory activity against the KB cancer cell line in vitro. In a previous contribution, we reported the isolation and structure determination of two prosapogenins (3 and 4) isolated from the butanol extract of the mild alkaline hydrolysate of the crude saponin fraction of the roots of Albizia adianthifolia (Schumach.) W. F. Wight (Mimo-saceae). 5 The further investigation of the saponins of this plant, obtained as a complex mixture, afforded two new acylated triterpene saponins, adianthifoliosides A (1) and B (2), from the 95% ethanolic extract of the roots of A. adianthifolia. This paper deals with the isolation and structure elucidation of these new acylated triterpene glycosides (1 and 2) and the evaluation of the immuno-modulatory activity of 1-4 (Chart 1) and the crude saponin mixture (CSM) on Jurkat T cell proliferation (human T cell leukemia) and the hemolytic activity of 1-4 on sheep erythrocytes. Results and Discussion The 95% ethanolic extract of the roots of A. adianthifolia was purified by precipitation with diethyl ether, yielding a crude saponin mixture, which was then dialyzed for 2 days. The powder obtained was submitted to column chromatography over Sephadex LH-20 and was separated by repeated medium-pressure liquid chromatography (MPLC) over normal Si gel, yielding compounds 1 and 2. Their structures were elucidated mainly by 1D and 2D NMR spectroscopy (COSY, TOCSY, NOESY, HSQC, and HMBC) and by FABMS and HRESIMS. Compound 1 was obtained as an amorphous powder. The HRESI mass spectrum (positive-ion mode) exhibited a quasimolecular ion peak at m/z 1714.7490 [M + Na] + (calcd 1714.7465), consistent with a molecular formula of C 79 H 121-NO 38. Upon acid hydrolysis with 2 N TFA at 120 °C, 1 afforded the aglycon 6, which was identified as acacic acid lactone (the 21,28-lactone derivative of acacic acid obtained under the experimental conditions used), by comparison of its NMR data with literature values. 6 The native aglycon was characterized as acacic acid from the 2D NMR spectra from 1. The sugars obtained from the saponin hydrolysate were identified as glucose, fucose, rhamnose, arabinose, and xylose by comparison with authentic samples. However , the presence of an N-acetamido group [IR 1639 and 1570 cm-1 ; 1 H NMR δ H 2.13 (3H, s, MeCO) and δ H 8.91 (1H, d, J) 8.8 Hz, NH); 13 C NMR δ C 22.9 and 171.9], together with the 1 H and 13 C NMR data of the C-2 of the Glc 1 (δ H 4.40 and δ C 56.8), suggested the presence of one 2-(acetylamino)-2-deoxyglucose (Glc 1 NAc) unit. In addition, alkaline hydrolysis of 1 afforded salicylic acid (SA, o-hydroxybenzoic acid, 7), identified by comparison of its 1 H and 13 C NMR data with those reported, 7 and a prosapoge-nin whose spectroscopic NMR data were in good agreement with those of 4. 5 The above data suggested that 1 is a 21-acyl-3,28-bisdesmoside. This was confirmed by the observation of glycosylation-and acylation-induced shifts in the 13 C NMR spectrum at δ C 88.7 (downfield shift of C-3), δ C 76.5 (downfield shift of C-21), and δ C 174.4 (upfield shift of C-28). The 1 H NMR spectrum of 1 displayed signals for seven anomeric protons at δ H 6.06 (br s), 5.83 (d, J) 7.5 Hz), 5.77 (br s), 5.10 (d, J) 7.5 Hz), 4.91 (d, J) 7.5 Hz), 4.90 (d, J) 7.8 Hz), and 4.79 (d, J) 8.0 Hz), which correlated with the carbon signals at δ C 109.8, 94.9, 101.2, 104.7, 103.8, 105.6, and 102.5, respectively, in the HSQC spectrum. Starting from the anomeric proton of each sugar unit, all the protons within each spin system were delineated using COSY with the aid of TOCSY and NOESY spectra. After assignments of the protons, the 13 C NMR resonances of each sugar unit were identified by HSQC and further confirmed by HMBC. The COSY and TOCSY spectr