Human DDX17 Unwinds Rift Valley Fever Virus Non-Coding RNAs

Rift Valley fever virus (RVFV) is a mosquito-transmitted virus from the Bunyaviridae family that causes high rates of mortality and morbidity in humans and ruminant animals. Previous studies indicated that DEAD-box helicase 17 (DDX17) restricts RVFV replication by recognizing two primary non-coding RNAs in the S-segment of the genome: the intergenic region (IGR) and 5′ non-coding region (NCR). However, we lack molecular insights into the direct binding of DDX17 with RVFV non-coding RNAs and information on the unwinding of both non-coding RNAs by DDX17. Therefore, we performed an extensive biophysical analysis of the DDX17 helicase domain (DDX17_135–555) and RVFV non-coding RNAs, IGR and 5’ NCR. The homogeneity studies using analytical ultracentrifugation indicated that DDX17_135–555, IGR, and 5’ NCR are pure. Next, we performed small-angle X-ray scattering (SAXS) experiments, which suggested that DDX17 and both RNAs are homogenous as well. SAXS analysis also demonstrated that DDX17 is globular to an extent, whereas the RNAs adopt an extended conformation in solution. Subsequently, microscale thermophoresis (MST) experiments were performed to investigate the direct binding of DDX17 to the non-coding RNAs. The MST experiments demonstrated that DDX17 binds with the IGR and 5’ NCR with a dissociation constant of 5.77 ± 0.15 µM and 9.85 ± 0.11 µM, respectively. As DDX17_135–555 is an RNA helicase, we next determined if it could unwind IGR and NCR. We developed a helicase assay using MST and fluorescently-labeled oligos, which suggested DDX17_135–555 can unwind both RNAs. Overall, our study provides direct evidence of DDX17_135–555 interacting with and unwinding RVFV non-coding regions.     

Human DDX3X Unwinds Japanese Encephalitis and Zika Viral 5′ Terminal Regions

Flavivirus genus includes many deadly viruses such as the Japanese encephalitis virus (JEV) and Zika virus (ZIKV). The 5′ terminal regions (TR) of flaviviruses interact with human proteins and such interactions are critical for viral replication. One of the human proteins identified to interact with the 5′ TR of JEV is the DEAD-box helicase, DDX3X. In this study, we in vitro transcribed the 5′ TR of JEV and demonstrated its direct interaction with recombinant DDX3X (K_d of 1.66 ± 0.21 µM) using microscale thermophoresis (MST). Due to the proposed structural similarities of 5′ and 3′ TRs of flaviviruses, we investigated if the ZIKV 5′ TR could also interact with human DDX3X. Our MST studies suggested that DDX3X recognizes ZIKV 5′ TR with a K_d of 7.05 ± 0.75 µM. Next, we performed helicase assays that suggested that the binding of DDX3X leads to the unwinding of JEV and ZIKV 5′ TRs. Overall, our data indicate, for the first time, that DDX3X can directly bind and unwind in vitro transcribed flaviviral TRs. In summary, our work indicates that DDX3X could be further explored as a therapeutic target to inhibit Flaviviral replication     

Electric field assisted solid-state interfacial joining of TaC-HfC ceramics without filler

The application of ultra-high-temperature ceramics (UHTCs) demands effective ways of joining in overcoming the problems associated with the fabrication of complex-shaped components. In this study, we choose to investigate a new method of rapidly joining pre-sintered TaC and HfC ceramics without any filler material using the spark plasma sintering (SPS) technique. A well-bonded TaC–HfC interface was observed with no apparent cracking and porosity at the joint. The joining mechanisms were predominantly driven by solid-state diffusion and localized plastic deformation. The nanomechanical properties of the TaC-HfC joint are better than the HfC while comparable to that of the TaC. High-load indentation (up to 200 N) results suggest that the TaC–HfC interface is stronger than the parent UHTCs with no crack propagating at the interface. Upon comparison with the parent UHTCs, the damaged area and the average crack length at the interface, reduced up to ~94% and ~56%, respectively. This study shows that the SPS technique can also apply to joining other UHTCs without any filler, resulting in the new field of developing complex components for the thermal protection system (TPS).     

Bridging organizations and strategic bridging functions in environmental governance and management

Strategic bridging functions performed by three voluntary, multi-stakeholder bridging organizations engaged in environmental governance and management in the Calgary Region of Alberta, Canada, are examined. Structured interviews revealed how the bridging organizations influenced an increase in municipal participation in regional-scale environmental governance and management activities. Bridging organizations connect stakeholders who would otherwise not be connected. They identify shared values and issues of concern, build trusting relationships, co-create knowledge, resolve conflict, and contribute to social learning processes. Social network structure is made visible through social network graphs. The legitimacy of natural resource management plans co-created through strategic bridging processes is also addressed.     

Nonylphenol affects gonadotropin levels in the pituitary gland and plasma of female rainbow trout

Female rainbow trout (Oncorhynchus mykiss) were exposed to 4-nonylphenol (NP) at (mean measured) concentrations of 0.7, 8.3, and 85.6 micrograms/L for 18 weeks, during early ovarian development. Fish were sampled sublethally every six weeks, and terminal samples were taken at 18 weeks. NP induced an estrogenic effect (the synthesis of vitellogenin) at concentrations of 8.3 and 85.6 micrograms/L. An effect on gonadotropin synthesis and secretion was also observed. Plasma follicle stimulating hormone (FSH) levels and FSH gene expression in the pituitary were the most sensitive endpoints assessed, being reduced at the lowest dose employed (0.7 microgram NP/L). Pituitary gland luteinizing hormone (LH) content was significantly lower in fish exposed to 85.6 micrograms NP/L, and LH gene expression was suppressed in fish exposed to 8.3 and 85.6 micrograms NP/L. In contrast, plasma LH concentration increased in these fish, but by a very minor absolute amount, and returned to control levels by the final sampling time. Gonadal development ceased in the fish exposed to 85.6 micrograms NP/L, and steroidogenesis in these fish was also markedly inhibited. Although the mechanisms underlying these responses are unknown, this study demonstrates that NP has adverse effects on pituitary function that can result in inhibition of ovarian development.     

A lakewide survey of Asian clam (Corbicula fluminea) distribution at warmwater discharges in Lake Michigan

The Asian clam Corbicula fluminea is among the most prolific aquatic invaders in the world; but in colder mid-latitude areas, like the Laurentian Great Lakes, their population expansion has likely been limited by poor overwinter survival. In these areas, Asian clams are typically found in thermal refugia like warmwater discharges from industrial facilities. We sought to identify the current extent of Asian clam populations in Lake Michigan and waters immediately adjacent to it, specifically at locations most likely to harbor overwintering populations – industrial warmwater discharges. During April–May 2017, we surveyed 17 locations around Lake Michigan. Evidence of Asian clam populations was found at four sites, though live specimens (n?=?3) were only found at the Indiana Harbor Ship Canal in East Chicago, IN. Shells or fragments of shells were found at Green Bay, WI, Waukegan, IL, and Port Sheldon, MI. Our findings indicate that although Asian clams are present in Lake Michigan, they are relatively rare, and remain isolated to a few small pockets of over-wintering habitat.     

Relative potencies and combination effects of steroidal estrogens in fish

The natural steroids estradiol-17beta (E2) and estrone (E1) and the synthetic steroid ethynylestradiol-17alpha (EE2) have frequently been measured in waters receiving domestic effluents. All of these steroids bind to the estrogen receptor(s) and have been shown to elicit a range of estrogenic responses in fish at environmentally relevant concentrations. At present, however, no relative potency estimates have been derived for either the individual steroidal estrogens or their mixtures in vivo. In this study the estrogenic activity of E2, E1, and EE2, and the combination effects of a mixture of E2 and EE2 (equi-potent fixed-ratio mixture), were assessed using vitellogenin induction in a 14-day in vivo juvenile rainbow trout screening assay. Median effective concentrations, relative to E2, for induction of vitellogenin were determined from the concentration-response curves and the relative estrogenic potencies of each of the test chemicals calculated. Median effective concentrations were between 19 and 26 ng L(-1) for E2, 60 ng L(-1) for E1, and between 0.95 and 1.8 ng L(-1) for EE2, implying that EE2 was approximately 11 to 27 times more potent than E2, while E2 was 2.3 to 3.2 times more potent than E1. The median effective concentration, relative to E2, for the binary mixture of E2 and EE2 was 15 ng L(-1) (comprising 14.4 ng L(-1) E2 and 0.6 ng L(-1) EE2). Using the model of concentration addition it was shown that this activity of the binary mixture could be predicted from the activity of the individual chemicals. The ability of each individual steroid to contribute to the overall effect of a mixture, even at individual no-effect concentrations, combined with the high estrogenic potency of the steroids, particularly the synthetic steroid EE2, emphasizes the need to consider the total estrogenic load of these chemicals in our waterways.     

Density,production, and survival of walleye (Sander vitreus) eggs in the Muskegon River,Michigan

Walleye (Sander vitreus) is an important sport fish in the Great Lakes that is experiencing low reproductive success after severe population declines starting in the late 1940s. In the Muskegon River, Michigan, natural reproduction of walleyes remains low and is largely supplemented by stocking. To determine factors influencing walleye reproductive success in the Muskegon River, we estimated walleye egg survival using insitu egg incubators covered with nitex screening (2003–2004) and estimated density and survival of fertilized eggs caught on furnace filter traps across different substrate types (2005–2006). We compared physical habitat suitability for walleye eggs under high and low flow scenarios. Density of walleye eggs was highest in regions of gravel/cobble substrates. Egg survival was higher in egg incubators (24–49.5%) than on furnace filter traps (2.0%), suggesting predation is an important source of walleye egg mortality in the Muskegon River. Cold water temperatures that extended developmental stage durations may also be an important source of egg mortality. The dynamic habitat suitability model predicted low suitability for eggs due to poor temperature and velocity conditions. Despite low egg survival rates, 40 million to 1 billion eggs were estimated to hatch. The low natural reproduction of walleyes in the Muskegon River is likely due to a combination of low walleye egg survival and failure of walleye larvae to reach their nursery grounds in Muskegon Lake.     

Evidence of successful river spawning by lake trout (Salvelinus namaycush) in the Lower Niagara River,Lake Ontario

Restoration of a wild-produced lake trout Salvelinus namaycush population in Lake Ontario has not been successful despite the adult population often meeting or exceeding restoration targets. Lack of high-quality spawning habitat in Lake Ontario is suggested as one impediment to recruitment of wild lake trout, although the quantity and location of spawning habitat is poorly understood. If high-quality spawning habitat is limited in Lake Ontario, lake trout may be using uncommon spawning locations such as rivers. Anecdotal angler accounts point to the Niagara River as a lake trout spawning location. To better understand the potential of the Niagara River as a spawning location, egg and juvenile fish collections were conducted 12–14 river kilometers from the mouth of the Niagara River from 2010 to 2012; and mature female lake trout with surgically implanted acoustic tags were monitored from 2015 to 2019. Genetic analyses confirmed 60% of collected eggs and 93% of collected post-hatch juvenile fish in the Niagara River were lake trout. Tagged female lake trout returned to the Niagara River over consecutive years during the spawning season. The short duration of lake trout presence in the river (mean = 56 days/year) suggests female lake trout use the Niagara River primarily for spawning. Diversity in spawning locations may provide lake trout population’s resilience against environmental variability through a portfolio effect. Improved identification of riverine spawning locations, including their overall contribution to wild recruitment, may be a useful tool for managers to restore a wild-produced population of lake trout in Lake Ontario.     

Using Survey Data and Mathematical Modeling to Prioritize Water Interventions in Developing Countries

Water Resources Management - A traditional cost-benefit analysis of potential water interventions in a given locality is a laborious and time-intensive process. To help decision makers identify...