Complete Genome Sequence of Mulberry Vein Banding Associated Virus, a New Tospovirus Infecting Mulberry.

Mulberry vein banding associated virus (MVBaV) that infects mulberry plants with typical vein banding symptoms had been identified as a tentative species of the genus Tospovirus based on the homology of N gene sequence to those of tospoviruses. In this study, the complete sequence of the tripartite RNA genome of MVBaV was determined and analyzed. The L RNA has 8905 nucleotides (nt) and encodes the putative RNA-dependent RNA polymerase (RdRp) of 2877 aa amino acids (aa) in the viral complementary (vc) strand. The RdRp of MVBaV shares the highest aa sequence identity (85.9%) with that of Watermelon silver mottle virus (WSMoV), and contains conserved motifs shared with those of the species of the genus Tospovirus. The M RNA contains 4731 nt and codes in ambisense arrangement for the NSm protein of 309 aa in the sense strand and the Gn/Gc glycoprotein precursor (GP) of 1,124 aa in the vc strand. The NSm and GP of MVBaV share the highest aa sequence identities with those of Capsicum chlorosis virus (CaCV) and Groundnut bud necrosis virus (GBNV) (83.2% and 84.3%, respectively). The S RNA is 3294 nt in length and contains two open reading frames (ORFs) in an ambisense coding strategy, encoding a 439-aa non-structural protein (NSs) and the 277-aa nucleocapsid protein (N), respectively. The NSs and N also share the highest aa sequence identity (71.1% and 74.4%, respectively) with those of CaCV. Phylogenetic analysis of the RdRp, NSm, GP, NSs, and N proteins showed that MVBaV is most closely related to CaCV and GBNV and that these proteins cluster with those of the WSMoV serogroup, and that MVBaV seems to be a species bridging the two subgroups within the WSMoV serogroup of tospoviruses in evolutionary aspect, suggesting that MVBaV represents a distinct tospovirus. Analysis of S RNA sequence uncovered the highly conserved 5'-/3'-ends and the coding regions, and the variable region of IGR with divergent patterns among MVBaV isolates.

Amikacin can be added to blood to reduce the fall in platelet count.

Our objective was to develop an effective method to prevent the fall in platelet count for patients with anticoagulant-dependent (AD) pseudothrombocytopenia, a spurious phenomenon due to anticoagulant-induced aggregation of platelets. We report a case of insidious multianticoagulant-dependent pseudothrombocytopenia in which AD pseudothrombocytopenia may be caused by 4 anticoagulants, eg, EDTA, sodium citrate, heparin, and sodium fluoride (NaF). Multianticoagulant-dependent pseudothrombocytopenia was confirmed by finding clumped platelets on microscopic evaluation in 4 anticoagulated blood samples. With this case, we tried a variety of reagents, including aminoglycosides, eg, gentamicin and amikacin, vitamin B(6), and aminophylline to inhibit pseudothrombocytopenia. Except for amikacin, all reagents failed to prevent pseudothrombocytopenia. Microscopic examination of K(2)-EDTA-, heparin-, sodium citrate-, and NaF-anticoagulated blood samples showed massive platelet clumping, but no aggregate was seen in the anticoagulated blood with amikacin. When amikacin was added within 1 hour after blood sample withdrawal, platelet, WBC, and RBC counts and hemoglobin level, mean corpuscular volume, and mean platelet volume remained unchanged for up to 4 hours at room temperature. These findings suggest that amikacin could inhibit and dissociate pseudo platelet aggregation in multianticoagulant-dependent pseudothrombocytopenia and EDTA-induced pseudothrombocytopenia.