Changes on the viral capsid surface during the evolution of porcine circovirus type 2 (PCV2) from 2009 till 2018 may lead to a better receptor binding

Porcine circovirus type 2 (PCV2) is the primary causative agent of porcine circovirus-associated diseases (PCVAD). Three major PCV2 genotypes (PCV2a, PCV2b, and PCV2d) have been identified globally. Despite their worldwide distribution, the prevalence and genetic evolution of PCV2 in Belgium has not previously been determined. In this study, 319 samples from animals suffering from diseases likely to be associated with PCV2 were collected from 2009 to 2018 and analysed by virus titration. The overall prevalence of PCV2 in PCVAD-suspected cases was 15.7 per cent (50/319). The phylogenetic analysis demonstrated that at least three genotypes (PCV2a, PCV2b, and PCV2d) circulated in Belgium from 2009 till 2018, and that PCV2 evolved from PCV2a to PCV2b and from PCV2d-1 to PCV2d-2. Sequence comparison among the forty-three PCV2 isolates showed that they had 89.7-100 per cent nucleotide-sequence and 88.5-100 per cent amino-acid-sequence identities. Three amino acid sites were under positive selection. Three-dimensional analysis of genotype-specific amino acids revealed that most of the mutations were on the outside of the cap protein with a few conserved mutations present on the inner side. Mutations toward more basic amino acids were found on the upper and tail parts of two connecting capsid proteins which form one big contact region, most probably involved in receptor binding. The lower part was relatively conserved. This polarity change together with the formation of an extruding part drive the virus to a more efficient GAG receptor binding. Taken together, these results showed a genotype shift from PCV2a to PCV2b and later on from PCV2d-1 to PCV2d-2, and a PCV2 evolution toward a better receptor binding capacity

The Architecture and Performance Evaluation of iSCSI-Based United Storage Network Merging NAS and SAN

With the ever increasing volume of data in networks, the traditional storage architecture is greatly challenged; more and more people pay attention to network storage. Currently, the main technology of network storage is represented by NAS (Network Attached Storage) and SAN (Storage Area Network). They are different, but mutually complementary and used under different circumstances; however, both NAS and SAN may be needed in the same company. To reduce the TOC (total of cost), for easier implementation, etc., people hope to merge the two technologies. Additionally, the main internetworking technology of SAN is the Fibre Channel; however, the major obstacles are in its poor interoperability, lack of trained staff, high implementation costs, etc. To solve the above-mentioned issues, this paper creatively introduces a novel storage architecture called USN (United Storage Networks), which uses the iSCSI to build the storage network, and merges the NAS and SAN techniques supplying the virtues and overcoming the drawbacks of both, and provides both file I/O and block I/O service simultaneously

Deglacial Variability of Antarctic Intermediate Water Penetration into the North Atlantic from Authigenic Neodymium Isotope Ratios

Understanding intermediate water circulation across the last deglacial is critical in assessing the role of oceanic heat transport associated with Atlantic Meridional Overturning Circulation variability across abrupt climate events. However, the links between intermediate water circulation and abrupt climate events such as the Younger Dryas (YD) and Heinrich Event 1 (H1) are still poorly constrained. Here, we reconstruct changes in Antarctic Intermediate Water (AAIW) circulation in the subtropical North Atlantic over the past 25 kyr by measuring authigenic neodymium isotope ratios in sediments from two sites in the Florida Straits. Our authigenic Nd isotope records suggest that there was little to no penetration of AAIW into the subtropical North Atlantic during the YD and H1. Variations in the northward penetration of AAIW into the Florida Straits documented in our authigenic Nd isotope record are synchronous with multiple climatic archives, including the Greenland ice core δ18O record, the Cariaco Basin atmosphere Δ14C reconstruction, the Bermuda Rise sedimentary Pa/Th record, and nutrient and stable isotope data from the tropical North Atlantic. The synchroneity of our Nd records with multiple climatic archives suggests a tight connection between AAIW variability and high-latitude North Atlantic climate change

Primary replication and invasion of the bovine gammaherpesvirus BoHV-4 in the genital mucosae

Bovine herpesvirus 4 (BoHV-4) is a gammaherpesvirus that is widespread in cattle. Ex vivo models with bovine genital tract mucosa explants were set up to study molecular/cellular BoHV-4-host interactions. Bovine posterior vagina, cervix and uterus body were collected from cows at two stages of the reproductive cycle for making mucosa explants. The BoHV-4 replication kinetics and characteristics within the three different mucosae of animals in the follicular and luteal phase were assessed by virus titration. The number of plaques, plaque latitude and number of infected cells were determined by immunofluorescence. BoHV-4 replicated in a productive way in all genital mucosal tissues. It infected single individual cells in both epithelium and lamina propria of the genital mucosae at 24 hours post-inoculation (hpi). Later, small BoHV-4 epithelial plaques were formed that did not spread through the basement membrane. 50% of the number of BoHV-4 infected cells were identified as cytokeratin(+) and CD172a(+) cells in the three parts of the genital tract at 24 hpi. Upon a direct injection of genital explants with BoHV-4, fibrocytes became infected, indicating that the unidentified 50% of the infected cells are most probably fibrocytes. In this study, in vivo-related in vitro genital tract models were successfully established and the early stage of the pathogenesis of a genital infection was clarified: BoHV-4 starts with a productive infection of epithelial cells in the reproductive tract, forming small foci followed by a non-productive infection of surveilling monocytic cells which help BoHV-4 to invade into deeper tissues

Gammaherpesvirus BoHV-4 infects bovine respiratory epithelial cells mainly at the basolateral side

Bovine herpesvirus 4 (BoHV-4) is a gammaherpesvirus that is widespread in cattle. However, only a few studies about the pathogenesis of BoHV-4 primary infection have been reported. In the present study, ex vivo models with bovine nasal and tracheal mucosa explants were used to study the cellular BoHV-4-host interactions. Infection was observed in nasal but not in tracheal epithelial cells. To find a possible correlation between the integrity and restricted infection of the respiratory epithelium, both nasal mucosal and tracheal explants were treated with EGTA, a drug that disrupts the intercellular junctions, before inoculation. The infection was analyzed based on the number of plaques, plaque latitude and number of infected single cells, as determined by immunofluorescence. BoHV-4 infection in nasal mucosal explants was enhanced upon opening the tight junctions with EGTA. Infection in tracheal explants was only found after treatment with EGTA. In addition, primary bovine respiratory epithelial cells (BREC) were isolated, grown at the air-liquid interface and infected either at the apical or basolateral side by BoHV-4. The results showed that BoHV-4 preferentially bound to and entered BREC at the basolateral surfaces of both nasal and tracheal epithelial cells. The percentage of BoHV-4 infection was significantly increased both from nasal and tracheal epithelial cells after treatment with EGTA, which indicates that the BoHV-4 receptor is mainly located at the basolateral surface of these cells. Thus, our findings demonstrate that integrity of the respiratory epithelium is crucial in the host's innate defense against primary BoHV-4 infections

Isotopic evidence for complex biogeochemical cycling of Cd in the eastern tropical South Pacific

Over the past decades, observations have confirmed decreasing oxygen levels and shoaling of oxygen minimum zones (OMZs) in the tropical oceans. Such changes impact the biogeochemical cycling of micronutrients such as Cd, but the potential consequences are only poorly constrained. Here, we present seawater Cd concentrations and isotope compositions for 12 depth profiles at coastal, nearshore and offshore stations from 4◦S to 14◦S in the eastern tropical South Pacific, where one of the world’s strongest OMZs prevails. The depth profiles of Cd isotopes display high δ114/110Cd at the surface and decreasing δ114/110Cd with increasing water depth, consistent with preferential utilization of lighter Cd isotopes during biological uptake in the euphotic zone and subsequent remineralization of the sinking biomass. In the surface and subsurface ocean, seawater displays similar δ114/110Cd signatures of 0.47 ± 0.23‰ to 0.82 ± 0.05‰ across the entire eastern tropical South Pacific despite highly variable Cd concentrations between 0.01 and 0.84 nmol/kg. This observation, best explained by an open system steady-state fractionation model, contrasts with previous studies of the South Atlantic and South Pacific Oceans, where only Cd-deficient waters have a relatively constant Cd isotope signature. For the subsurface to about 500 m depth, the variability of seawater Cd isotope compositions can be modeled by mixing of remineralized Cd with subsurface water from the base of the mixed layer. In the intermediate and deep eastern tropical South Pacific (>500 m), seawater [Cd] and δ114/110Cd appear to follow the distribution and mixing of major water masses. We identified modified AAIW of the ETSP to be more enriched in [Cd] than AAIW from the source region, whilst both water masses have similar δ114/110Cd. A mass balance estimate thus constrains a δ114/110Cd of between 0.38‰ and 0.56‰ for the accumulated remineralized Cd in the ETSP. Nearly all samples show a tight coupling of Cd and PO4 concentrations, whereby surface and deeper waters define two distinct linear trends. However, seawater at a coastal station located within a pronounced plume of H2S, is depleted in [Cd] and features significantly higher δ114/110Cd. This signature is attributed to the formation of authigenic CdS with preferential incorporation of lighter Cd isotopes. The process follows a Rayleigh fractionation model with a fractionation factor of α114/110Cdseawater-CdS = 1.00029. Further deviations from the deep Cd–PO4 trend were observed for samples with O2 < 10 μmol/kg and are best explained by in situ CdS precipitation within the decaying organic matter even though dissolved H2S was not detectable in ambient seawater

Preferential use of Siglec-1 or Siglec-10 by type 1 and type 2 PRRSV strains to infect PK15S1-CD163 and PK15S10-CD163 cells

Cellular entry mediators define whether the cell is permissive to PRRSV infection. Porcine sialoadhesin (pSn, Siglec-1) and CD163 are main entry mediators facilitating infection of porcine macrophages by PRRSV. Recently, Siglec-10 was demonstrated to be an alternative receptor for PRRSV. To examine if virulence and pathogenicity of PRRSV strains could be correlated with the use of different Siglecs, a PK15 cell line recombinantly expressing Siglec-1 and CD163 (PK15(S1-CD163)) and a PK15 cell line recombinantly expressing Siglec-10 and CD163 (PK15(S10-CD163)) were used to compare the virus replication of 7 genotype 1 subtype 1 strains (G1s1), 2 genotype 1 subtype 3 (G1s3) strains and 5 genotype 2 (G2) strains. Some strains (08VA (G1s1), 13V117 (G1s1), 17V035 (G1s1), VR2332 (G2)) were poor virus producers (= 10(6) TCID50/mL. PK15(S10-CD163) cells exhibited a higher efficiency in virus production per infected cell than the PK15(S1-CD163) cells. The G1s1 strains LV and 07V063 infected more cells in the PK15(S1-CD163), whereas the 94V360 and 08VA strains preferred PK15(S10-CD163). The highly virulent G1s3 strains Lena and Su1-Bel showed a strong preference for PK15(S1-CD163). The G2 strains MN-184, SDSU-73, Korea17 had a much higher infection rate in PK15(S10-CD163), while the reference strain VR2332 and the NADC30 strain had a slight preference for -PK15(S1-CD163). Differences in receptor use may influence the outcome of a PRRSV infection in pigs and explain in part the virulence/pathogenicity of PRRSV strains

Pingmu Decoction Induces Orbital Preadipocytes Apoptosis In Vitro

Pingmu Decoction is the Traditional Chinese Medicine which has treated Graves’ Ophthalmopathy (GO) in the inactive stage for more than ten years. This study was to explore the mechanism of Pingmu Decoction of inhibiting preadipocytes in GO patients from differentiating into mature adipocytes. Human orbital preadipocytes were isolated and cultured through tissue explant method. Orbital preadipocytes were induced into mature adipocytes. The medicinal serum was prepared from rats. The cells were treated with medicinal serum which were divided into three groups, low dose group (5%), medium dose group (10%), and high dose group (20%). The cells viabilities were observed by Oil Red O staining, MTT method, and Annexin V/propidium iodide (PI) double staining. Effect of Pingmu Decoction on cell apoptosis rate of orbital matured adipocytes was measured by flow cytometry. The genes Fas and Fas L from cell groups were tested by RT-PCR and Western blotting. The expression of master adipogenic transcription factors, including peroxisome proliferation-activity receptor (PPAR) γ and CCAAT/enhancer binding protein (C/EBP) α, was tested by Western blotting. Pingmu Decoction could reduce orbital preadipocytes viability and induce apoptosis of mature adipocyte via Fas/Fas L signaling pathway. Pingmu Decoction reduced lipid accumulation and downregulated the expression of PPAR γ and C/EBP α. Pingmu Decoction may play a therapeutic effect by reducing the accumulation of orbital adipocytes

Effects of 238U variability and physical transport on water column 234Th downward fluxes in the coastal upwelling system off Peru

The eastern boundary region of the southeastern Pacific Ocean hosts one of the world's most dynamic and productive upwelling systems with an associated oxygen minimum zone (OMZ). The variability in downward export fluxes in this region, with strongly varying surface productivity, upwelling intensities and water column oxygen content, is however poorly understood. Thorium-234 (234Th) is a powerful tracer to study the dynamics of export fluxes of carbon and other elements, yet intense advection and diffusion in nearshore environments impact the assessment of depth-integrated 234Th fluxes when not properly evaluated. Here we use vessel-mounted acoustic Doppler current profiler (VmADCP) current velocities, satellite wind speed and in situ microstructure measurements to determine the magnitude of advective and diffusive fluxes over the entire 234Th flux budget at 25 stations from 11 to 16∘ S in the Peruvian OMZ. Contrary to findings along the GEOTRACES P16 eastern section, our results showed that weak surface wind speed during our cruises induced low upwelling rates and minimal upwelled 234Th fluxes, whereas vertical diffusive 234Th fluxes were important only at a few shallow shelf stations. Horizontal advective and diffusive 234Th fluxes were negligible because of small alongshore 234Th gradients. Our data indicated a poor correlation between seawater 238U activity and salinity. Assuming a linear relationship between the two would lead to significant underestimations of the total 234Th flux by up to 40 % in our study. Proper evaluation of both physical transport and variability in 238U activity is thus crucial in coastal 234Th flux studies. Finally, we showed large temporal variations on 234Th residence times across the Peruvian upwelling zone and cautioned future carbon export studies to take these temporal variabilities into consideration while evaluating carbon export efficiency