Time-dependent effects of microplastics on soil bacteriome.

Microplastic threats to biodiversity, health and ecological safety are adding to concern worldwide, but the real impacts on the functioning of organisms and ecosystems are obscure owing to their inert characteristics. Here we investigated the long-lasting ecological effects of six prevalent microplastic types: polyethylene (PE), polypropylene (PP), polyamide (PA), polystyrene (PS), polyethylene terephthalate (PET), and polyvinyl chloride (PVC) on soil bacteria at a 2 % (w/w) level. Due to the inertia and lack of available nitrogen of these microplastics, their effects on bacteriome tended to converge after one year and were strongly different from their short-term effects. The soil volumes around microplastics were very specific, in which the microplastic-adapted bacteria (e.g., some genera in Actinobacteria) were enriched but the phyla Bacteroidetes and Gemmatimonadetes declined, resulting in higher microbial nitrogen requirements and reduced organic carbon mineralization. The reshaped bacteriome was specialized in the genetic potential of xenobiotic and lipid metabolism as well as related oxidation, esterification, and hydrolysis processes, but excessive oxidative damage resulted in severe weakness in community genetic information processing. According to model predictions, microplastic effects are indirectly derived from nutrients and oxidative stress, and the effects on bacterial functions are stronger than on structure, posing a heavy risk to soil ecosystems.

[Effects of radiation changes on net ecosystem exchange of carbon dioxide in a middle subtropical Chinese fir plantation]. / 辐射变化对中亚热带杉木人工林净CO2交换的影响.

Total solar radiation is an important factor affecting carbon exchange in forest ecosystem. In order to understand the effects of radiation change on carbon exchange in Chinese fir plantation, long-term monitoring data of carbon dioxide flux and meteorological factors measured by open eddy covariance system and meteorological gradient observation system were used in this study. The clearness index (kt) was used to represent the condition of solar radiation. We analyzed the effects of kt on net ecosystem exchange of carbon dioxide (NEE) in the central subtropical Chinese fir plantation during the growing season (from April to October). The results showed that total solar radiation in clear sky was usually higher in the morning than that in the afternoon, and that NEE was lower in the morning than in the afternoon. Such difference in NEE reached the maximum when the solar elevation angle was about 50°. At the medium kt(0.42-0.52), carbon absorption of Chinese fir plantation was the strongest. The ave-rage maximum relative change of NEE in 10 years in different solar elevation angles ranged from 11.0% to 29.4%, while the minimum and maximum critical values appeared at 35°-40° and 45°-50°, respectively. When kt was at the moderate degree due to the existence of clouds, carbon absorption and diffuse photosynthetically active radiation of Chinese fir plantation reached the maximum, and the latter might be the main reason for the former. Moderate radiation condition with the presence of cloud clould promote NEE of Chinese fir plantation and lead to largest carbon absorption.

Amino acid mutations in the env gp90 protein that modify N-linked glycosylation of the Chinese EIAV vaccine strain enhance resistance to neutralizing antibodies.

The Chinese EIAV vaccine is an attenuated live virus vaccine obtained by serial passage of a virulent horse isolate (EIAVL) in donkeys (EIAVD) and, subsequently, in donkey cells in vitro. In this study, we compare the env gene of the original horse virulent virus (EIAVL) with attenuated strains serially passaged in donkey MDM (EIAVDLV) and donkey dermal cells (EIAVFDDV). Genetic comparisons among parental and attenuated strains found that vaccine strains contained amino acid substitutions/deletions in gp90 that resulted in a loss of three potential N-linked glycosylation sites, designated g5, g9, and g10. To investigate the biological significance of these changes, reverse-mutated viruses were constructed in the backbone of the EIAVFDDV infectious molecular clone (pLGFD3). The resulting virus stocks were characterized for replication efficiency in donkey dermal cells and donkey MDM, and were tested for sensitivity to neutralization using sera from two ponies experimentally infected with EIAVFDDV. Results clearly show that these mutations generated by site-directed mutagenesis resulted in cloned viruses with enhanced resistance to serum neutralizing antibodies that were also able to recognize parental viruses. This study indicates that these mutations played an important role in the attenuation of the EIAV vaccine strains.

AKT signalling and mitochondrial pathways are involved in mushroom polysaccharide-induced apoptosis and G1 or S phase arrest in human hepatoma cells.

This study describes molecular mechanisms for inhibiting tumour cell proliferation using polysaccharides from medicinal mushrooms in human hepatoma cells. The results show that regarding cell cycle-related proteins, three types of polysaccharides significantly enhance the expression of p27(Kip) in HepG2 and Bel-7404 cells, while suppressing the activity of cyclin D1/CDK4 and/or cyclin E/CDK2. Considering apoptosis-related factors, the polysaccharides suppressed AKT activity through the inhibition of AKT phosphorylation at Thr(308) and/or Ser(473). The growth of HepG2 and Bel-7404 cells was suppressed by the up-regulation of a subunit of PI3K and phospho-PTEN, which are modulators of AKT activity. The polysaccharides also activated the mitochondria-mediated apoptosis pathway by stimulating the activation of Bcl-2 family proteins to release cytochrome c and Smac and cleave caspase-9 and caspase-3 in HepG2 and Bel-7404 cells. These factors have a potent effect on cell cycle arrest in G(1) and/or S phase and induce apoptosis in HepG2 and Bel-7404 cells.

Amino acid mutations in the env gp90 protein that modify N-linked glycosylation of the Chinese EIAV vaccine strain enhance resistance to neutralizing antibodies.

The Chinese EIAV vaccine is an attenuated live-virus vaccine obtained by serial passage of a virulent horse isolate (EIAV(L)) in donkeys (EIAV(D)), and subsequently in donkey cells in vitro. In this study, we compare the env gene of the original horse virulent virus (EIAV(L)) with attenuated strains serially passaged in donkey MDM (EIAV(DLV)), and donkey dermal cells (EIAV(FDDV)). Genetic comparisons among parental and attenuated strains found that vaccine strains contained amino acid substitutions/deletions in gp90 that resulted in a loss of three potential N-linked glycosylation sites, designated g5, g9, and g10. To investigate the biological significance of these changes, reverse-mutated viruses were constructed in the backbone of the EIAV(FDDV) infectious molecular clone (pLGFD3). The resulting virus stocks were characterized for replication efficiency in donkey dermal cells and donkey MDM, and were tested for sensitivity to neutralization using sera from two ponies experimentally infected with EIAV(FDDV). The results clearly show that these mutations generated by site-directed mutagenesis resulted in cloned viruses with enhanced resistance to serum-neutralizing antibodies that were also able to recognize parental viruses. The results of this study indicate that these mutations play an important role in the attenuation of the EIAV vaccine strains.

In vivo evolution of the gp90 gene and consistently low plasma viral load during transient immune suppression demonstrate the safety of an attenuated equine infectious anemia virus (EIAV) vaccine.

To study the in vivo evolution of the attenuated Chinese equine infectious anemia virus (EIAV) vaccine, viral gp90 gene variation and virus replication in immunosuppressed hosts were investigated. The results showed that after vaccination, the gp90 gene followed an evolutionary trend of declining diversity. The trend coincided with the maturation of immunity to EIAV, and eventually, the gp90 gene became highly homologous. The sequences of these predominant quasispecies were consistently detected up to 18 months after vaccination. Furthermore, after transient immune suppression with dexamethasone, the plasma viral RNA copy number of the vaccine strain in three vaccinated ponies remained consistently below the "pathogenic threshold" level, while the viral load increased by 25,000-fold in the positive control of an inapparent carrier of the parental virulent strain. This study is the first to provide evidence for the safety of an attenuated lentiviral vaccine with decreased genomic diversity and consistently low viral replication under suppressed immunity.

Genetic variation in the long terminal repeat associated with the transition of Chinese equine infectious anemia virus from virulence to avirulence.

A highly virulent strain of equine infectious anemia virus (EIAV) lost its fatal virulence but retained the desired antigens during serial passage over 130 generations in leukocytes in vitro. We compared the long terminal repeat (LTR) sequences of the different generations and found that three stable genetic variations occurred in the transcriptional start site, the initial base of TAR, and the pre-mRNA cleavage site at the R-U5 boundary, respectively. These three mutations happened at the inflexion of virus pathogenicity loss; therefore, the function of these mutations needs to be further addressed.

[Construction of an infectious clone of equine infectious anemia virus by N-glycosylation reverse-mutations].

To elucidate the role of N-glycosylation in fetal donkey dermal cell (FDD)-attenuated equine infectious anemia virus (EIAV), we constructed an N-glycosylation reverse-mutation molecular clone, pLGN191N236N246. This viral molecular clone was derived from the infectious clone pLGFD3-8 by site-directed mutagenesis. This clone was used to transfect fetal donkey dermal (FDD) cells. Infectious characteristics of transfectants were monitored by RT-PCR, indirect immune fluorescence and reverse transcriptase activity assay. After three passages in FDD cells, viral replications in the supernatant of cell cultures were detected by all the above three methods and viral particles were clearly observed by electron microscopy. The construction of the N-glycosylation reverse-mutation infectious clone provides a solid basis for further study of the mechanism of attenuated pathogenesis and increased immune protection of EIAV attenuated vaccines.