Tula virus: a newly detected hantavirus carried by European common voles.

A novel hantavirus has been discovered in European common voles, Microtus arvalis and Microtus rossiaemeridionalis. According to sequencing data for the genomic RNA S segment and nucleocapsid protein and data obtained by immunoblotting with a panel of monoclonal antibodies, the virus, designated Tula virus, is a distinct novel member of the genus Hantavirus. Phylogenetic analyses of Tula virus indicate that it is most closely related to Prospect Hill, Puumala, and Muerto Canyon viruses. The results support the view that the evolution of hantaviruses follows that of their primary carriers. Comparison of strains circulating within a local rodent population revealed a genetic drift via accumulation of base substitutions and deletions or insertions. The Tula virus population from individual animals is represented by quasispecies, indicating the potential for rapid evolution of the agent.     

APP intracellular domain formation and unaltered signaling in the presence of familial Alzheimer's disease mutations

One of the cardinal neuropathological findings in brains from Alzheimer's disease (AD) patients is the occurrence of amyloid beta-peptide (Abeta) deposits. The gamma-secretase-mediated intramembrane proteolysis event generating Abeta also results in the release of the APP intracellular domain (AICD), which may mediate nuclear signaling. It was recently shown that AICD starts at a position distal to the site predicted from gamma-secretase cleavage within the membrane. This novel site, the epsilon site, is located close to the inner leaflet of the membrane bilayer. The relationship between proteolysis at the gamma and epsilon sites has not been fully characterized. Here we studied AICD signaling in intact cells using a chimeric C99 molecule and a luciferase reporter system. We show that the release of AICD from the membrane takes place in a compartment downstream of the endoplasmic reticulum, is dependent on presenilin proteins, and can be inhibited by treatment with established gamma-secretase inhibitors. Moreover, we find that AICD signaling remains unaltered from C99 derivatives containing mutations associated with increased Abeta42 production and familial AD. These findings indicate that there are very similar routes for Abeta and AICD formation but that FAD-linked mutations in APP primarily affect gamma-secretase-mediated Abeta42 formation, and not AICD signaling.     

Cross-protection against challenge with Puumala virus after immunization with nucleocapsid proteins from different hantaviruses

Hantaviruses are rodent-borne agents that cause hemorrhagic fever with renal syndrome or hantavirus pulmonary syndrome in humans. The nucleocapsid protein (N) is relatively conserved among hantaviruses and highly immunogenic in both laboratory animals and humans, and it has been shown to induce efficient protective immunity in animal models. To investigate the ability of recombinant N (rN) from different hantaviruses to elicit cross-protection, we immunized bank voles with rN from Puumala (PUUV), Topografov (TOPV), Andes (ANDV), and Dobrava (DOBV) viruses and subsequently challenged them with PUUV. All animals immunized with PUUV and TOPV rN were completely protected. In the group immunized with DOBV rN, 7 of 10 animals were protected, while only 3 of 8 animals were protected in the group immunized with ANDV rN, which is more closely related to PUUV rN than DOBV rN. Humoral and cellular immune responses after rN immunization were also investigated. The highest cross-reactive humoral responses against PUUV antigen were detected in sera from ANDV rN-immunized animals, followed by those from TOPV rN-immunized animals, and only very low antibody cross-reactivity was observed in sera from DOBV rN-immunized animals. In proliferation assays, T lymphocytes from animals immunized with all heterologous rNs were as efficiently recalled in vitro by PUUV rN as were T lymphocytes from animals immunized with homologous protein. In summary, this study has shown that hantavirus N can elicit cross-protective immune responses against PUUV, and the results suggest a more important role for the cellular arm of the immune response than for the humoral arm in cross-protection elicited by rN.     

Effects of Propagule Phenology (Non-Dormant Versus Dormant) and Planting System (Vertical Versus Horizontal) on Growth Performance of Willow Clones Grown Under Different Weeding Regimes

To assess the effects of propagule phenology and planting system on growth performance of three willow clones grown under different weeding regimes, a field experiment was performed in central Sweden 2014–2016. Freshly harvested (non-dormant) and cold-stored (dormant) cuttings (planted vertically) and billets (planted horizontally) from willow clones Tordis, Tora, and Jorr were planted in weeded and in unweeded plots. Sprouting was significantly higher for willows grown from non-dormant (74%) than dormant (58%) propagules and for cuttings (84%) compared with billets (42%). Survival was higher for willows from non-dormant propagules in weeded (71%) compared with unweeded (63%) plots, willows from dormant propagules in weeded (72%) compared with unweeded (60%) plots, and for willows from cuttings (93%) compared with billets (39%). During 2014–2016, aboveground biomass production was significantly higher for willows from cuttings (11.71 t DW ha^?1) than from billets (6.13 t DW ha^?1), grown in weeded (15.29 t DW ha^?1) than in unweeded (2.55 t DW ha^?1) plots, and differed significantly among willow clones (11.48, 9.27, and 6.01 t DW ha^?1 for Tordis, Tora, and Jorr, respectively). In this study, (i) planting with cold-stored and freshly harvested willow propagules was equally successful and therefore cold storage could be potentially avoided and replaced with planting of freshly harvested propagules in early spring; however, (ii) in terms of measured growth performance parameters, willows grown from cuttings performed better than grown from billets; and (iii) weed competition significantly reduced survival and aboveground biomass production, confirming that weed control during establishment of willow is crucial.     

Ex Vivo Stability of the Rodent-Borne Hantaan Virus in Comparison to That of Arthropod-Borne Members of the Bunyaviridae Family

The possible effect of virus adaptation to different transmission routes on virus stability in the environment is not well known. In this study we have compared the stabilities of three viruses within the Bunyaviridae family: the rodent-borne Hantavirus Hantaan virus (HTNV), the sand fly-borne Phlebovirus sandfly fever Sicilian virus (SFSV), and the tick-borne Nairovirus Crimean-Congo hemorrhagic fever virus (CCHFV). These viruses differ in their transmission routes: SFSV and CCHFV are vector borne, whereas HTNV is spread directly between its hosts, and to humans, via the environment. We studied whether these viruses differed regarding stability when kept outside of the host. Viral survival was analyzed at different time points upon exposure to different temperatures (4°C, 20°C, and 37°C) and drying at 20°C. We observed clearly different stabilities under wet conditions, particularly at 4°C, where infectious SFSV, HTNV, and CCHFV were detectable after 528, 96, and 15 days, respectively. All three viruses were equally sensitive to drying, as shown by drying on aluminum discs. Furthermore, HTNV and SFSV partially survived for 2 min in 30% ethanol, whereas CCHFV did not. Electron microscopy images of HTNV, SSFSV, and CCHFV stored at 37°C until infectivity was lost still showed the occurrence of virions, but with abnormal shapes and densities compared to those of the nonincubated samples. In conclusion, our study points out important differences in ex vivo stability among viruses within the Bunyaviridae family.     

Wild-Type Puumala Hantavirus Infection Induces Cytokines, C-Reactive Protein, Creatinine, and Nitric Oxide in Cynomolgus Macaques

Hantaviruses cause two severe human diseases: hemorrhagic fever with renal syndrome (HFRS) and hantavirus pulmonary syndrome (HPS). Approximately 200,000 cases are reported annually, and there is to date no specific treatment available. A major obstacle in studying the medical aspects of HFRS and HPS has been the lack of an adequate animal model. Here we show that infection of cynomolgus macaques by wild-type Puumala hantavirus resulted in typical signs of HFRS including lethargy, anorexia, proteinuria, and/or hematuria, in addition to cytokine (interleukin 6 [IL-6], IL-10, and tumor necrosis factor alpha), C-reactive protein, creatinine, and nitric oxide responses. Viral RNA was detected in plasma from days 3 to 7 postinoculation until days 24 to 28 postinoculation, infectious virus was recovered, and the virus-specific immune responses (immunoglobulin M [IgM], IgG, and neutralizing antibodies) mimicked those seen in humans. The results indicated that the monkey model will provide a valuable tool for studies of pathogenesis, candidate vaccines, and antivirals for hantavirus disease.     

Neutralizing Human Fab Fragments against Measles Virus Recovered by Phage Display

Five human recombinant Fab fragments (Fabs) specific for measles virus (MV) proteins were isolated from three antibody phage display libraries generated from RNAs derived from bone marrow or splenic lymphocytes from three MV-immune individuals. All Fabs reacted in an enzyme-linked immunosorbent assay with MV antigens. In radioimmunoprecipitation assays two of the Fabs, MV12 and MT14, precipitated an approximately equal 80-kDa protein band corresponding to the hemagglutinin (H) protein from MV-infected Vero cell cultures, while two other Fabs, MT64 and GL29, precipitated an approximately equal 60-kDa protein corresponding the nucleocapsid (N) protein. In competition studies with MV fusion, H- and N protein-specific monoclonal antibodies (MAbs), the H-specific Fabs predominantly blocked the binding of H-specific MAbs, while the N-specific Fabs blocked MAbs to N. In addition, N-specific Fabs bound to denatured MV N protein in Western blotting. The specificity of the fifth Fab, MV4, could not be determined. By plaque reduction assays, three of the five Fabs, MV4, MV12, and MT14, exhibited neutralizing activity (80% cutoff) against MV (LEC-KI strain) at concentrations ranging between approximately 2 and 7 microg x ml(-1). Neutralization capacity against MV strains Edmonston and Schwarz was also detected, albeit at somewhat higher Fab concentrations. In conclusion, three neutralizing Fabs were isolated, two of them reactive against the H glycoprotein of MV and another reactive against an undefined epitope. This is the first study in which MV-neutralizing human recombinant Fab antibodies have been isolated from phage display libraries.     

Effects of Cutting Phenology (Non-dormant Versus Dormant) on Early Growth Performance of Three Willow Clones Grown Under Different Weed Treatments and Planting Dates

To assess the effects of cutting phenology on early growth performance of three willow clones grown under different weed treatments and planting dates, freshly harvested (non-dormant) and cold-stored (dormant) cuttings from willow clone Tora, Jorr, and Olof were planted in bucket experiment outdoors in central Sweden on five planting dates (May–June 2013) with or without a model weed (spring barley). Non-dormant cuttings sprouted faster than dormant cuttings when planted early in the season. For cuttings planted later in the season, bud sprouting was affected only by willow clone. Aboveground biomass production was affected by cutting phenology, planting date, clone, and weed treatment. When planted on May 3 and May 10, biomass produced from non-dormant and dormant cuttings did not differ, while willows grown from dormant cuttings produced 59% more aboveground biomass than willows grown from non-dormant cuttings when planted on May 24–June 16. Tora produced on average 12% more biomass than Jorr and Olof, and weed competition reduced aboveground biomass production on average with 36%. The ability of willow to suppress weeds (WSA) was 26 (non-dormant cuttings) and 12% (dormant cuttings) higher for willows planted on May 3 compared with WSA of willows grown from cuttings planted later in the season. The ability to tolerate competition from weeds (WT) was 51 and 52% lower for willows grown from non-dormant and dormant cuttings planted late in the season compared with WT of willows planted earlier in the season. We conclude that planting with long-term cold storage of willow cuttings can be replaced with planting freshly harvested cuttings when planting is performed in early season, and that weed competition strongly reduces biomass production. Weed control during the establishment phase is crucial in order to maximize willow biomass production.     

Morphological studies of the stromal changes after artificial induction of the decidual reaction in rats

Summary Stromal changes during early oil-induced decidualization were studied. Rats previously sensitized with estradiol after an experimental delay were used. Among the initial stromal characteristics that developed after induction, the following sequence was found: ultrastructural alterations of stromal cell nucleoli, extracellular edema, and Pontamine Blue Reaction (PBR). The results suggest that stromal cell differentiation into decidual cells is initiated before the PBR becomes visible. This is in accordance with biochemical and morphological findings at blastocyst implantation. The appearance of an edema before the PBR furthermore suggests that also the increase in vascular permeability is initiated early and takes place slowly. The functional relationships between the vascular and stromal cell changes are, however, not determinable with the method used.This work was supported by grants from the Medical Faculty at the University of Uppsala and the Swedish Medical Research Council (Project No 12X-70 to O. Nilsson)Technical assistance: Mrs Marianne Ljungkvist and Miss Kerstin Rystedt