protocols-for-investigating-host-tissue-distribution-transmission

Protocols for Investigating the Host-tissue Distribution, Transmission-mode, and Effect on the Host Fitness of a Densovirus in the Cotton Bollworm

Many novel viruses have been discovered in animal hosts using next-generation sequencing technologies. Previously, we reported a mutualistic virus, ...

Chinese Academy of Agricultural Sciences

Spinal Muscular Atrophy (SMA) is a neurodegenerative disease that is caused by deletion of the SMN (Survival of Motor Neuron) gene. The SMN protein is essential for cell survival and co-localized with TIA-1/R and G3BP, two characteristic markers of stress granules (SGs). To further study the SMN function in stress granules and in response to stress, we generated stable cell lines with SMN knockdown. Our data indicate that suppression of SMN drastically reduces cellular ability to form stress granules in response to stress treatment. In addition, we show that SMN deficiency sensitizes cells to sodium arsenite and H(2)O(2), two well-known stress inducers, leading to cell death at a much lower concentration of inducers in SMN knockdown cells than in control cells. Interestingly, the cell death is correlated with formation of stress granules, suggesting that involvement of SMN in formation of stress granules may play an important role in cell survival. Furthermore, rescue of SGs formation by overexpression of G3BP can reverse the defective formation of stress granules and results in partial abrogation of cell death against SMN deficiency. We deduce that modulation of stress response may be useful for potential SMN treatment.

SMN deficiency reduces cellular ability to form stress granules, sensitizing cells to stress.

HCHs and DDTs were banned in 1983 in China; however, they are still remaining in various environmental media. Since endosulfan was introduced in China in 1994, it is widely used in agriculture. In this study, temporal and spatial uses of endosulfan, HCHs, and DDTs in Gansu province of China have been presented. It is estimated that the total usage is 701Â tons for endosulfan between 1994 and 2007, 1,712Â tons for HCHs between 1952 and 1983, and 462Â tons for DDTs between 1951 and 1983, respectively. Endosulfan usage increased dramatically in 1998 due to its application on other crops except on cotton. The HCH and DDT usage displayed a rapid increase after 1972, reaching the peak in 1976 and in 1975, respectively; since then, they declined until being banned in 1983. The gridded usage inventories of these three kinds of organochlorine pesticides in Gansu province, with a 1/4Â° longitude by 1/6Â° latitude resolution, have been created by using different crops for endosulfan and the area of dry farmland for HCHs and DDTs as surrogate data. The most intensive use was in northwestern regions for endosulfan and southeastern regions for HCHs and DDTs in Gansu province.

Gridded inventories of historical usage for selected organochlorine pesticides in Gansu Province, China.

Neutrophil-to-lymphocyte ratio (NLR) and platelet-to-lymphocyte ratio (PLR) have recently been investigated as two new inflammatory markers used in the assessment of systemic inflammation in many diseases. The purpose of the study was to investigate their relation with disease activity in newly diagnosed SLE patients.

Neutrophil-to-lymphocyte ratio (NLR) and platelet-to-lymphocyte ratio (PLR) were associated with disease activity in patients with systemic lupus erythematosus.

The complete genome sequence of a novel single-stranded RNA virus in Nesidiocoris tenuis was determined by RNA-seq and rapid amplification of cDNA ends (RACE) methodologies and was named N. tenuis virus 1. The genomic RNA was 3970 nucleotides (nt) in length and contained two putative open reading frames (ORFs). ORF1 encoded a polypeptide with 283 amino acids containing a viral (superfamily 1) RNA helicase (Hel) domain, and ORF2 encoded a polypeptide with 294 amino acids containing an RNA-dependent RNA polymerase (RdRP) domain. Phylogenetic analysis using the deduced amino acid sequences indicated that the N. tenuis virus 1 clustered with Blackford virus; however, the low bootstrap values and unique genomic structure suggested that the virus is a prototype of a new type of unclassified viruses. The prevalence of N. tenuis virus 1 infection in field populations of N. tenuis differed between three locations, with 28.32% of the 113 sampled individuals testing positive for the virus.

A novel single-stranded RNA virus in Nesidiocoris tenuis.

The complete genome of a novel virus found in Adelphocoris suturalis was determined by RNA-seq and named Adelphocoris suturalis-associated virus 1 (ASV1). ASV1 has a single-stranded RNA genome of 10,845 nucleotides in length and contains five putative open reading frames (ORFs). ORF1 encodes a polypeptide of 2592 amino acids (aa) and contains four conserved domains: a viral RNA methyltransferase domain, an FtsJ-like methyltransferase domain, a viral RNA helicase domain and an RNA-dependent RNA polymerase domain. ORF2, ORF3, ORF4 and ORF5 encode polypeptides of 190, 461, 103 and 159 aa, respectively, of which only ORF5 contains a conserved domain, the Tobacco mosaic virus-coat superfamily. Phylogenetic analysis with the deduced amino acid sequences indicated that ASV1 clusters with the Drosophila-related Boutonnet virus. The similar genomic structure and high bootstrap value identified in the maximum-likelihood tree suggest that ASV1 (possibly alongside Boutonnet virus) could be considered the prototype of a new taxon of unclassified insect viruses. The prevalence of ASV1 infection in wild populations of A. suturalis was at a low level (6.60%, 14 positives from 212 samples).

The genome sequence of a novel RNA virus in Adelphocoris suturalis.

The cotton bollworm, Helicoverpa armigera, is one of the most important agricultural pests of many economic crops worldwide. Herein, we found a novel single-strand RNA virus by RNA-Seq and Polymerase Chain Reaction (PCR) method in H. armigera named Helicoverpa armigera iflavirus (HaIV), which possessed a genome with 10,017 nucleotides in length and contained a single large open reading frame (ORF) encoding a putative polyprotein of 3021 amino acids with a predicted molecular mass of 344.16kDa and a theoretical isoelectric point (pI) of 6.45. The deduced amino acid sequence showed highest similarity (61.0%) with the protein of Lymantria dispar Iflavirus 1. Phylogenetic analysis with putative RdRp amino acid sequences indicated that the virus clustered with members of the genus Iflavirus. The virus was mainly distributed in the fat body of its host and was found to be capable of both horizontal and vertical transmission. The efficiency of perorally horizontal transmission was dose dependent (100% infection rate with a viral dose of 10(8)copies/μl) while vertical transmission efficiency was found to be relatively low (<28.57%). These results suggest that we have found a novel member of genus Iflavirus in H. armigera.

Characterization of a novel member of genus Iflavirus in Helicoverpa armigera.

Herein, we report the identification of putative promoters for the non-structural proteins (NS) and capsid structural proteins (VP) of Helicoverpa armigera densovirus (HaDV2) as well as a potential mechanism for how these promoters might be regulated. For the first time, we report that VP is able to transactivate the VP promoter and, to a lesser degree, the NS promoter in densoviruses. In addition to this, another promoter-like sequence designated P2, when co-transfected with the VP gene, enhanced luciferase activity by approximately 35 times compared to a control. This suggests that there are two promoters for VP in HaDV2 and that the VP of parvoviruses might play a more important role in viral transcription than previously appreciated.

Xianming Yang

State Key Laboratory for Biology of Plant Diseases and Insect Pests,
Institute of Plant Protection,
State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection

Chinese Academy of Agricultural Sciences

Protocols for Investigating the Host-tissue Distribution, Transmission-mode, and Effect on the Host Fitness of a Densovirus in the Cotton Bollworm

Xianming Yang

State Key Laboratory for Biology of Plant Diseases and Insect Pests,Institute of Plant Protection,State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection

Chinese Academy of Agricultural Sciences

Protocols for Investigating the Host-tissue Distribution, Transmission-mode, and Effect on the Host Fitness of a Densovirus in the Cotton Bollworm

State Key Laboratory for Biology of Plant Diseases and Insect Pests,
Institute of Plant Protection,
State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection