Association of Eriophyes dimocarpi (Acari: Eriophyidae) with longan witches’ broom disease in Vietnam

Longan witches’ broom (LgWB) syndrome was observed in all longan-growing provinces of Vietnam. Key symptoms include: small, stunted shoots with curved, rolled up margins, deformed leaves with blisters and hairy patches on the underside; abnormal development of flower structures, flower abortion, failure to produce fruits or developing small fruits. During 2012–2015, field surveys and study of the relationship between LgWB symptoms and Eriophyes dimocarpi mites presence were carried out. Moreover, molecular analysis, electron microscopy observations, E. dimocarpi inoculations, pruning of affected spikelets and miticide experiments were performed. An average of 36.58% of longan seedlings inoculated with E. dimocarpi mites showed typical symptoms of LgWB at about 27.5 days after inoculation. The use of several pesticides in different combinations showed high efficacy on E. dimocarpi mites elimination and of symptomatology incidence reduction. These results suggest a significant association of E. dimocarpi with LgWB syndrome in Vietnam.     

Spontaneous Ejaculation in a Wild Indo-Pacific Bottlenose Dolphin (Tursiops aduncus)

Spontaneous ejaculation, which is defined as the release of seminal fluids without apparent sexual stimulation, has been documented in boreoeutherian mammals. Here we report spontaneous ejaculation in a wild Indo-Pacific bottlenose dolphin (Tursiops aduncus), and present a video of this rare behavior. This is the first report of spontaneous ejaculation by an aquatic mammal, and the first video of this behavior in animals to be published in a scientific journal.     

Préparation et contrôle de qualité de la solution injectable de gallium-68 édotréotide (DOTATOC (68Ga))

Radiopharmacists of the Nuclear Medicine department of hospital Tenon have prepared and controlled the ⁶⁸Ga-labeled DOTATOC, for 4 years, as part of a clinical research study. The aim of this article is to share our experience, since this activity is not yet developed in France. Radiolabelling of DOTATOC (⁶⁸Ga) requires the settling of a ⁶⁸Ge/⁶⁸Ga generator, which is connected to an automated PC-controlled radiopharmaceutical labelling device (Elusynth ⁶⁸Ga, Iason) and comprises several steps. Performed quality controls (QC) are those commonly used for radiopharmaceuticals including: appearance, pH, radiochemical purity (RCP), radionuclide purity (PRN) and determination of the physical half-life. Bacterial endotoxins and sterility tests are systematically done. We obtained a mean value of radiolabelling yield around 45%. The results of QC are always in accordance with the specifications. The preparation failed in 7% of the 195 DOTATOC (⁶⁸Ga), over the last 4 years. It is important to note that the preparation of DOTATOC (⁶⁸Ga) monopolizes the radiopharmacist during 3 hours. This radiolabelling technique can be easily applied to other peptides, in order to develop other ⁶⁸Ga-labelled PET tracers.     

Discovery of loperamide as an antagonist of angiopoietin1 and angiopoietin2 by virtual screening

The angiopoietin-Tie2 binding and related signal transduction pathways are crucial for vascular angiogenesis, blood vessel integrity and maturation. In this study, we preformed a virtual screening of small molecules targeting to Tie2. The binding site was selected at the extracellular ligand binding region of Tie2, rather than its conventional endocellular ATP binding region. It was found that loperamide, a widely-used antidiarrhea drug, was among the top hits. The binding between loperamide and Tie2 was confirmed by surface plasmon resonance (SPR) assay. Loperamide competitively inhibited the binding of both angiopoietin1 and angiopoietin2. These results indicate that loperamide is an antagonist of angiopoietin1 and angiopoietin2.     

Establishment of a novel permissive cell line for the propagation of hepatitis C virus by expression of microRNA miR122

The robust cell culture systems for hepatitis C virus (HCV) are limited to those using cell culture-adapted clones (HCV in cell culture [HCVcc]) and cells derived from the human hepatoma cell line Huh7. However, accumulating data suggest that host factors, including innate immunity and gene polymorphisms, contribute to the variation in host response to HCV infection. Therefore, the existing in vitro systems for HCV propagation are not sufficient to elucidate the life cycle of HCV. A liver-specific microRNA, miR122, has been shown to participate in the efficient replication of HCV. In this study, we examined the possibility of establishing a new permissive cell line for HCV propagation by the expression of miR122. A high level of miR122 was expressed by a lentiviral vector placed into human liver cell lines at a level comparable to the endogenous level in Huh7 cells. Among the cell lines that we examined, Hep3B cells stably expressing miR122 (Hep3B/miR122) exhibited a significant enhancement of HCVcc propagation. Surprisingly, the levels of production of infectious particles in Hep3B/miR122 cells upon infection with HCVcc were comparable to those in Huh7 cells. Furthermore, a line of "cured" cells, established by elimination of HCV RNA from the Hep3B/miR122 replicon cells, exhibited an enhanced expression of miR122 and a continuous increase of infectious titers of HCVcc in every passage. The establishment of the new permissive cell line for HCVcc will have significant implications not only for basic HCV research but also for the development of new therapeutics.     

Adaptation of a duck influenza A virus in quail

Quail are thought to serve as intermediate hosts of influenza A viruses between aquatic birds and terrestrial birds, such as chickens, due to their high susceptibility to aquatic-bird viruses, which then adapt to replicate efficiently in their new hosts. However, does replication of aquatic-bird influenza viruses in quail similarly result in their efficient replication in humans? Using sialic acid-galactose linkage-specific lectins, we found both avian (sialic acid-α2-3-galactose [Siaα2-3Gal] linkages on sialyloligosaccharides)- and human (Siaα2-6Gal)-type receptors on the tracheal cells of quail, consistent with previous reports. We also passaged a duck H3N2 virus in quail 19 times. Sequence analysis revealed that eight mutations accumulated in hemagglutinin (HA) during these passages. Interestingly, many of the altered HA amino acids found in the adapted virus are present in human seasonal viruses, but not in duck viruses. We also found that stepwise stalk deletion of neuraminidase occurred during passages, resulting in reduced neuraminidase function. Despite some hemagglutinin mutations near the receptor binding pocket, appreciable changes in receptor specificity were not detected. However, reverse-genetics-generated viruses that possessed the hemagglutinin and neuraminidase of the quail-passaged virus replicated significantly better than the virus possessing the parent HA and neuraminidase in normal human bronchial epithelial cells, whereas no significant difference in replication between the two viruses was observed in duck cells. Further, the quail-passaged but not the original duck virus replicated in human bronchial epithelial cells. These data indicate that quail can serve as intermediate hosts for aquatic-bird influenza viruses to be transmitted to humans.     

Crystal structure of Hfq from Bacillus subtilis in complex with SELEX-derived RNA aptamer: insight into RNA-binding properties of bacterial Hfq

Bacterial Hfq is a protein that plays an important role in the regulation of genes in cooperation with sRNAs. Escherichia coli Hfq (EcHfq) has two or more sites that bind RNA(s) including U-rich and/or the poly(A) tail of mRNA. However, functional and structural information about Bacillus subtilis Hfq (BsHfq) including the RNA sequences that specifically bind to it remain unknown. Here, we describe RNA aptamers including fragment (AG)₃A that are recognized by BsHfq and crystal structures of the BsHfq–(AG)₃A complex at 2.2 Å resolution. Mutational and structural studies revealed that the RNA fragment binds to the distal site, one of the two binding sites on Hfq, and identified amino acid residues that are critical for sequence-specific interactions between BsHfq and (AG)₃A. In particular, R32 appears to interact with G bases in (AG)₃A. Poly(A) also binds to the distal site of EcHfq, but the overall RNA structure and protein–RNA interaction patterns engaged in the R32 residues of BsHfq–(AG)₃A differ from those of EcHfq–poly(A). These findings provide novel insight into how the Hfq homologue recognizes RNA.