Toxoplasma MIC2 Is a Major Determinant of Invasion and Virulence

Like its apicomplexan kin, the obligate intracellular protozoan Toxoplasma gondii actively invades mammalian cells and uses a unique form of gliding motility. The recent identification of several transmembrane adhesive complexes, potentially capable of gripping external receptors and the sub-membrane actinomyosin motor, suggests that the parasite has multiple options for host-cell recognition and invasion. To test whether the transmembrane adhesin MIC2, together with its partner protein M2AP, participates in a major invasion pathway, we utilized a conditional expression system to introduce an anhydrotetracycline-responsive mic2 construct, allowing us to then knockout the endogenous mic2 gene. Conditional suppression of MIC2 provided the first opportunity to directly determine the role of this protein in infection. Reduced MIC2 expression resulted in mistrafficking of M2AP, markedly defective host-cell attachment and invasion, the loss of helical gliding motility, and the inability to support lethal infection in a murine model of acute toxoplasmosis. Survival of mice infected with MIC2-deficient parasites correlated with lower parasite burden in infected tissues, an attenuated inflammatory immune response, and induction of long-term protective immunity. Our findings demonstrate that the MIC2 protein complex is a major virulence determinant for Toxoplasma infection and that MIC2-deficient parasites constitute an effective live-attenuated vaccine for experimental toxoplasmosis

Class II ADP-ribosylation factors are required for efficient secretion of Dengue viruses

This article is available open access through the publisher’s website.Identification and characterization of virus-host interactions are very important steps toward a better understanding of the molecular mechanisms responsible for disease progression and pathogenesis. To date, very few cellular factors involved in the life cycle of flaviviruses, which are important human pathogens, have been described. In this study, we demonstrate a crucial role for class II Arf proteins (Arf4 and Arf5) in the dengue flavivirus life cycle. We show that simultaneous depletion of Arf4 and Arf5 blocks recombinant subviral particle secretion for all four dengue serotypes. Immunostaining analysis suggests that class II Arf proteins are required at an early pre-Golgi step for dengue virus secretion. Using a horseradish peroxidase protein fused to a signal peptide, we show that class II Arfs act specifically on dengue virus secretion without altering the secretion of proteins through the constitutive secretory pathway. Co-immunoprecipitation data demonstrate that the dengue prM glycoprotein interacts with class II Arf proteins but not through its C-terminal VXPX motif. Finally, experiments performed with replication-competent dengue and yellow fever viruses demonstrate that the depletion of class II Arfs inhibits virus secretion, thus confirming their implication in the virus life cycle, although data obtained with West Nile virus pointed out the differences in virus-host interactions among flaviviruses. Our findings shed new light on a molecular mechanism used by dengue viruses during the late stages of the life cycle and demonstrate a novel function for class II Arf proteins.Research Fund for Control of Infectious Diseases of Hong Kong and BNP Paribas Corporate and Investment Banking

Cirmtuzumab inhibits Wnt5a-induced Rac1 activation in chronic lymphocytic leukemia treated with ibrutinib.

Signaling via the B cell receptor (BCR) plays an important role in the pathogenesis and progression of chronic lymphocytic leukemia (CLL). This is underscored by the clinical effectiveness of ibrutinib, an inhibitor of Bruton's tyrosine kinase (BTK) that can block BCR-signaling. However, ibrutinib cannot induce complete responses (CR) or durable remissions without continued therapy, suggesting alternative pathways also contribute to CLL growth/survival that are independent of BCR-signaling. ROR1 is a receptor for Wnt5a, which can promote activation of Rac1 to enhance CLL-cell proliferation and survival. In this study, we found that CLL cells of patients treated with ibrutinib had activated Rac1. Moreover, Wnt5a could induce Rac1 activation and enhance proliferation of CLL cells treated with ibrutinib at concentrations that were effective in completely inhibiting BTK and BCR-signaling. Wnt5a-induced Rac1 activation could be blocked by cirmtuzumab (UC-961), an anti-ROR1 mAb. We found that treatment with cirmtuzumab and ibrutinib was significantly more effective than treatment with either agent alone in clearing leukemia cells in vivo. This study indicates that cirmtuzumab may enhance the activity of ibrutinib in the treatment of patients with CLL or other ROR1+ B-cell malignancies

Identifying Asthma genetic signature patterns by mining Gene Expression BIG Datasets using Image Filtering Algorithms

Asthma is a treatable but incurable chronic inflammatory disease affecting more than 14% of the UAE population. Asthma is still a clinical dilemma as there is no proper clinical definition of asthma, unknown definitive underlying mechanisms, no objective prognostic tool nor bedside noninvasive diagnostic test to predict complication or exacerbation. Big Data in the form of publicly available transcriptomics can be a valuable source to decipher complex diseases like asthma. Such an approach is hindered by technical variations between different studies that may mask the real biological variations and meaningful, robust findings. A large number of datasets of gene expression microarray images need a powerful tool to properly translate the image intensities into truly differential expressed genes between conditioned examined from the noise. Here we used a novel bioinformatic method based on the coefficient of variance to filter nonvariant probes with stringent image analysis processing between asthmatic and healthy to increase the power of identifying accurate signals hidden within the heterogeneous nature of asthma. Our analysis identified important signaling pathways members, namely NFKB and TGFB pathways, to be differentially expressed between severe asthma and healthy controls. Those vital pathways represent potential targets for future asthma treatment and can serve as reliable biomarkers for asthma severity. Proper image analysis for the publicly available microarray transcriptomics data increased its usefulness to decipher asthma and identify genuine differentially expressed genes that can be validated across different datasets

Biologie de reproduction chez Thaumatococcus Daniellii (benn.) Benth. et hook. (marantaceae) en Côte D’ivoire

Le processus de reproduction de Thaumatococcus daniellii a été étudié en vue d'une domestication à but économique. Ainsi, l'identification des animaux contribuant à la pollinisation a été réalisée à partir d'observations effectuées dans un dispositif expérimental réglementant l'accès différentiel des animaux aux parcelles. Une pollinisation manuelle suivie de l'analyse des fruits, par rapport à la teneur engraines, ont été effectuées. Les résultats ont révélé l'intervention d'agents extérieurs dans le processus de pollinisation, impliquant un phénomène d'allogamie chez cette plante. Ces agents extérieurs qui agissent en synergie sont Nectarinia olivacea (oiseau) et Aphis sp. (puceron). La prépondérance des fruits contenant une graine,l'échec élevé de l'autopollinisation et la prédominance d'une l'allopollinisation traduisent une incompatibilité génétique dans la régulation de la pollinisation des fleurs chez cette plante

Interactions between organic friction modifier additives

The interactions of different additives in engine oils can create synergistic or antagonistic effects. This paper studies how mixing different organic friction modifier additives affects friction reducing properties of lubricants in the boundary lubrication regime. Amines of different degree of saturation were mixed with either glycerol monooleate (GMO) or oleic acid in hexadecane. The model lubricants thus formed were characterised with Fourier-transform infrared spectroscopy. Friction tests in reciprocating motion using ball-on-disc steel-steel contacts were conducted to examine the tribological performance of these lubricants. Worn surfaces were examined using X-ray photoelectron spectroscopy. Oleic acid and oleylamine, a primary amine. Were found to form a partial ionic liquid, providing synergistic friction reduction. This positive interaction reduces with increasing degree of saturation of the amine. No synergistic effect was observed between GMO and oleylamine,suggesting that GMO does not hydrolyse into a carboxylic acid within a rubbing contact in the presence of amine. Keywords: Boundary Lubrication, Additives, Friction Abbreviations: organic friction modifier (OFM); glycerol monooleate (GMO); ionic liquid (IL); oleylamine (OA); diocylamine (DA); trihexylamine (TA); dimethylhexadecaylamine (DM16); high frequency reciprocating rig (HFRR); X-ray photoelectron spectroscopy (XPS)

Accretion, Primordial Black Holes and Standard Cosmology

Primordial Black Holes evaporate due to Hawking radiation. We find that the evaporation time of primordial black holes increase when accretion of radiation is included.Thus depending on accretion efficiency more and more number of primordial black holes are existing today, which strengthens the idea that the primordial black holes are the proper candidate for dark matter.Comment: 11 pages, 3 figure

Expression, Purification, and Biophysical Characterization of a Secreted Anthrax Decoy Fusion Protein in Nicotiana benthamiana.

Anthrax toxin receptor-mediated drug development for blocking anthrax toxin action has received much attention in recent decades. In this study, we produced a secreted anthrax decoy fusion protein comprised of a portion of the human capillary morphogenesis gene-2 (CMG2) protein fused via a linker to the fragment crystallizable (Fc) domain of human immunoglobulin G1 in Nicotiana benthamiana plants using a transient expression system. Using the Cauliflower Mosaic Virus (CaMV) 35S promoter and co-expression with the p19 gene silencing suppressor, we were able to achieve a high level of recombinant CMG2-Fc-Apo (rCMG2-Fc-Apo) protein accumulation. Production kinetics were observed up to eight days post-infiltration, and maximum production of 826 mg/kg fresh leaf weight was observed on day six. Protein A affinity chromatography purification of the rCMG2-Fc-Apo protein from whole leaf extract and apoplast wash fluid showed the homodimeric form under non-reducing gel electrophoresis and mass spectrometry analysis confirmed the molecular integrity of the secreted protein. The N-glycosylation pattern of purified rCMG2-Fc-Apo protein was analysed; the major portion of N-glycans consists of complex type structures in both protein samples. The most abundant (>50%) N-glycan structure was GlcNAc₂(Xyl)Man₃(Fuc)GlcNAc₂ in rCMG2-Fc-Apo recovered from whole leaf extract and apoplast wash fluid. High mannose N-glycan structures were not detected in the apoplast wash fluid preparation, which confirmed the protein secretion. Altogether, these findings demonstrate that high-level production of rCMG2-Fc-Apo can be achieved by transient production in Nicotiana benthamiana plants with apoplast targeting

Outbreak of West Nile virus causing severe neurological involvement in children, Nuba Mountains, Sudan, 2002.

An atypical outbreak of West Nile virus (WNV) occurred in Ngorban County, South Kordophan, Sudan, from May to August 2002. We investigated the epidemic and conducted a case-control study in the village of Limon. Blood samples were obtained for cases and controls. Patients with obvious sequelae underwent cerebrospinal fluid (CSF) sampling as well. We used enzyme-linked immunosorbent assay (ELISA) and neutralization tests for laboratory diagnosis and identified 31 cases with encephalitis, four of whom died. Median age was 36 months. Bivariate analysis did not reveal any significant association with the risk factors investigated. Laboratory analysis confirmed presence of IgM antibodies caused by WNV in eight of 13 cases, indicative of recent viral infection. The unique aspects of the WNW outbreak in Sudan, i.e. disease occurrence solely among children and the clinical domination of encephalitis, involving severe neurological sequelae, demonstrate the continuing evolution of WNV virulence. The spread of such a virus to other countries or continents cannot be excluded