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91.
92.
Stokes KL Chi MH Sakamoto K Newcomb DC Currier MG Huckabee MM Lee S Goleniewska K Pretto C Williams JV Hotard A Sherrill TP Peebles RS Moore ML 《Journal of virology》2011,85(12):5782-5793
Airway mucus is a hallmark of respiratory syncytial virus (RSV) lower respiratory tract illness. Laboratory RSV strains differentially induce airway mucus production in mice. Here, we tested the hypothesis that RSV strains differ in pathogenesis by screening six low-passage RSV clinical isolates for mucogenicity and virulence in BALB/cJ mice. The RSV clinical isolates induced variable disease severity, lung interleukin-13 (IL-13) levels, and gob-5 levels in BALB/cJ mice. We chose two of these clinical isolates for further study. Infection of BALB/cJ mice with RSV A2001/2-20 (2-20) resulted in greater disease severity, higher lung IL-13 levels, and higher lung gob-5 levels than infection with RSV strains A2, line 19, Long, and A2001/3-12 (3-12). Like the line 19 RSV strain, the 2-20 clinical isolate induced airway mucin expression in BALB/cJ mice. The 2-20 and 3-12 RSV clinical isolates had higher lung viral loads than laboratory RSV strains at 1 day postinfection (p.i.). This increased viral load correlated with higher viral antigen levels in the bronchiolar epithelium and greater histopathologic changes at 1 day p.i. The A2 RSV strain had the highest peak viral load at day 4 p.i. RSV 2-20 infection caused epithelial desquamation, bronchiolitis, airway hyperresponsiveness, and increased breathing effort in BALB/cJ mice. We found that RSV clinical isolates induce variable pathogenesis in mice, and we established a mouse model of clinical isolate strain-dependent RSV pathogenesis that recapitulates key features of RSV disease. 相似文献
93.
Mário C BarrosoJúnior Guilherme P Esteves Thiago P Nunes Lucia MG Silva Alvaro CD Faria Pedro L Melo 《Biomedical engineering online》2011,10(1):14
Introduction
A novel system that combines a compact mobile instrument and Internet communications is presented in this paper for remote evaluation of tremors. The system presents a high potential application in Parkinson's disease and connects to the Internet through a TCP/IP protocol. Tremor transduction is carried out by accelerometers, and the data processing, presentation and storage were obtained by a virtual instrument. The system supplies the peak frequency (fp), the amplitude (Afp) and power in this frequency (Pfp), the total power (Ptot), and the power in low (1-4 Hz) and high (4-7 Hz) frequencies (Plf and Phf, respectively). 相似文献94.
Saman Bowatte Paul CD Newton Shona Brock Phil Theobald Dongwen Luo 《The ISME journal》2015,9(1):265-267
Nitrous oxide (N2O) emissions from grazed pastures are a product of microbial transformations of nitrogen and the prevailing view is that these only occur in the soil. Here we show this is not the case. We have found ammonia-oxidising bacteria (AOB) are present on plant leaves where they produce N2O just as in soil. AOB (Nitrosospira sp. predominantly) on the pasture grass Lolium perenne converted 0.02–0.42% (mean 0.12%) of the oxidised ammonia to N2O. As we have found AOB to be ubiquitous on grasses sampled from urine patches, we propose a ‘plant'' source of N2O may be a feature of grazed grassland.In terms of climate forcing, nitrous oxide (N2O) is the third most important greenhouse gas (Blunden and Arndt, 2013). Agriculture is the largest source of anthropogenic N2O (Reay et al., 2012) with about 20% of agricultural emissions coming from grassland grazed by animals (Oenema et al., 2005).Grazed grassland is a major source of N2O because grazers harvest nitrogen (N) from plants across a wide area but recycle it back onto the pasture, largely as urine, in patches of very high N concentration. The N in urine patches is often in excess of what can be used by plants resulting in losses through leaching as nitrate, as N2O and through volatilisation as ammonia (NH3) creating a high NH3 environment in the soil and plant canopy; an important point that we will return to later. The established wisdom is that N2O is generated exclusively by soil-based microbes such as ammonia-oxidising bacteria (AOB). This soil biology is represented in models designed to simulate N2O emissions and the soil is a target for mitigation strategies such as the use of nitrification inhibitors.We have previously shown that pasture plants can emit N2O largely through acting as a conduit for emissions generated in the soil, which are themselves controlled to some degree by the plant (Bowatte et al., 2014). In this case the origin of the emission is still the soil microbes. However, AOB have been found on the leaves of plants, for example, Norway spruce (Papen et al., 2002; Teuber et al., 2007) and weeds in rice paddies (Bowatte et al., 2006), prompting us to ask whether AOB might be present on the leaves of pasture species and contribute to N2O emissions as they do in soil.We looked for AOB on plants in situations where NH3 concentrations were likely to be high, choosing plants from urine patches in grazed pastures and plants from pastures surrounding a urea fertiliser manufacturing plant. DNA was extracted from the leaves (including both the surface and apoplast) and the presence of AOB tested using PCR. AOB were present in all the species we examined—the grasses Lolium perenne, Dactylis glomerata, Anthoxanthum odoratum, Poa pratensis, Bromus wildenowii and legumes Trifolium repens and T. subterraneum.To measure whether leaf AOB produce N2O, we used intact plants of ryegrass (L. perenne) lifted as cores from a paddock that had been recently grazed by adult sheep. The cores were installed in a chamber system designed to allow sampling of above- and belowground environments separately (Bowatte et al., 2014). N2O emissions were measured from untreated (control) plants and from plants where NH3 was added to the aboveground chamber and leaves were either untreated or sterilised by wiping twice with paper towels soaked in 1% hypoclorite (Sturz et al., 1997) and then with sterile water. We tested for the presence and abundance of AOB on the leaves by extracting DNA and using PCR and real-time PCR targeting the ammonia monoxygenase A (amoA) gene, which is characteristic of AOB. AOB identity was established using cloning and DNA sequencing. Further details of these experiments can be found in the Supplementary Information.The addition of NH3 to untreated plants significantly stimulated N2O emissions (P<0.001) compared with the controls; by contrast, the plants with sterilised leaves produced significantly less N2O than controls (P<0.001) even with NH3 added (Figure 1) providing strong evidence for emissions being associated with bacteria on the leaves. Control plants did emit N2O suggesting there was either sufficient NH3 available for bacterially generated emissions and/or other plant-based mechanisms were involved (Bowatte et al., 2014).Open in a separate windowFigure 1Effect of an elevated NH3 atmosphere and surface sterilisation of leaves on leaf N2O emissions measured over 1-h periods on three occasions during the day. Values are means (s.e.m.), where n=7.The major AOB species identified was Nitrosospira strain III7 that has been previously shown to produce N2O (Jiang and Bakken, 1999). We measured 109 AOB cells per m2 ryegrass leaf, assuming a specific leaf area of 250 cm2 g−1 leaf.The rate of production of N2O (0.1–0.17 mg N2O-N per m2 leaf area per hour) can be translated to a field situation using the leaf area index (LAI)—1 m2 leaf per m2 ground would be an LAI of 1. LAI in a pasture can vary from <1 to >6 depending on the management (for example, Orr et al., 1988). At LAI of 1, the AOB leaf emission rate would equate to a N2O emission rate of about 0.1–0.3 mg N2O-N per m2 ground per hour. By comparison, the emission rates measured after dairy cattle urine (650 kg N ha−1) was applied to freely and poorly drained soil were 0.024–1.55 and 0.048–3.33 mg N2O-N per m2 ground per hour, respectively (Li and Kelliher, 2005).The fraction of the NH3 that was converted to N2O by the leaf AOB was 0.02–0.42% (mean 0.12%). The mean value is close to that measured for Nitrosospira strains including strain III7 isolated from acidic, loamy and sandy soils where values ranged from 0.07 to 0.10% (Jiang and Bakken, 1999). This is good evidence that the AOB on leaves have the capacity to produce N2O at the same rate as AOB in soils. We do not suggest that leaf AOB will produce as much N2O as soil microbes; however, because leaf AOB have access to a source of substrate—volatilised NH3—that is unavailable to soil microbes and may constitute 26% (Laubach et al., 2013) to 40% (Carran et al., 1982) of the N deposited in the urine, N2O emissions from these aboveground AOB are additional to soil emissions. Further research is required to identify the situations in which leaf AOB contribute to total emissions and to quantify this contribution. 相似文献
95.
Anderson MO Sherrill J Madrid PB Liou AP Weisman JL DeRisi JL Guy RK 《Bioorganic & medicinal chemistry》2006,14(2):334-343
A parallel synthetic strategy to the 9-aminoacridine scaffold of the classical anti-malarial drug quinacrine (2) is presented. The method features a new route to 9-chloroacridines that utilizes triflates of salicylic acid derivatives, which are commercially available in a variety of substitution patterns. The route allows ready variation of the two diversity elements present in this class of molecules: the tricyclic aromatic heterocyclic core, and the disubstituted diamine sidechain. In this study, a library of 175 compounds was designed, although only 93 of the final products had purities acceptable for screening. Impurity was generally due to incomplete removal of 9-acridones (18), a degradation product of the 9-chloroacridine synthetic intermediates. The library was screened against two strains of Plasmodium falciparum, including a model of the drug-resistant parasite, and six novel compounds were found to have IC(50) values in the low nanomolar range. 相似文献
96.
Peter F. Galbreath Nathan D. Adams Lee. W. Sherrill III Thomas H. Martin 《Environmental Biology of Fishes》2006,75(2):183-193
Synopsis An effect of ploidy on thermal tolerance in juvenile trout was assessed in a series of tests comparing time to chronic lethal
maximum (CLMax). Diploid and triploid fish were produced from a common spawn for three different groups each of brook trout
Salvelinus fontinalis and of rainbow trout Oncorhynchus mykiss. One or two CLMax tests were performed per group, on between 15 and 50 individuals per ploidy within groups. The tests involved
exposure of fish to a progressive 2°C day−1 water temperature increase and recording of the time at which each individual fish reached loss of equilibrium (LE). The
time to LE data were rank transformed and analyzed as a randomized complete block design. Although relative performance varied
among trials, the analysis indicated overall differences due to ploidy were small and nonsignificant among both brook trout
and rainbow trout. Size proved to be significantly correlated with time to LE in the brook trout trials, but not in the rainbow
trout trials. Two of the six groups included a large proportion of fish which had received a heat shock following fertilization,
but were not successfully triploidized. In both cases, thermal tolerance of the heat-shocked diploids was similar to that
of the non-heat shocked control diploids, indicating no persistent effect of the heat shock on thermal tolerance. 相似文献
97.
Kumaran Kandasamy Sujatha S Mohan Rajesh Raju Shivakumar Keerthikumar Ghantasala S Sameer Kumar Abhilash K Venugopal Deepthi Telikicherla Daniel J Navarro Suresh Mathivanan Christian Pecquet Sashi Kanth Gollapudi Sudhir Gopal Tattikota Shyam Mohan Hariprasad Padhukasahasram Yashwanth Subbannayya Renu Goel Harrys KC Jacob Jun Zhong Raja Sekhar Vishalakshi Nanjappa Lavanya Balakrishnan Roopashree Subbaiah YL Ramachandra Abdul B Rahiman Keshava TS Prasad Jian-Xin Lin Jon CD Houtman Stephen Desiderio Jean-Christophe Renauld Stefan N Constantinescu Osamu Ohara Toshio Hirano Masato Kubo Sujay Singh Purvesh Khatri Sorin Draghici Gary D Bader Chris Sander Warren J Leonard Akhilesh Pandey 《Genome biology》2010,11(1):1-9
98.
Taylor PB Stewart FP Dunnington DJ Quinn ST Schulz CK Vaidya KS Kurali E Lane TR Xiong WC Sherrill TP Snider JS Terpstra ND Hertzberg RP 《Journal of biomolecular screening》2000,5(4):213-226
The transition from manual to robotic high throughput screening (HTS) in the last few years has made it feasible to screen hundreds of thousands of chemical entities against a biological target in less than a month. This rate of HTS has increased the visibility of bottlenecks, one of which is assay optimization. In many organizations, experimental methods are generated by therapeutic teams associated with specific targets and passed on to the HTS group. The resulting assays frequently need to be further optimized to withstand the rigors and time frames inherent in robotic handling. Issues such as protein aggregation, ligand instability, and cellular viability are common variables in the optimization process. The availability of robotics capable of performing rapid random access tasks has made it possible to design optimization experiments that would be either very difficult or impossible for a person to carry out. Our approach to reducing the assay optimization bottleneck has been to unify the highly specific fields of statistics, biochemistry, and robotics. The product of these endeavors is a process we have named automated assay optimization (AAO). This has enabled us to determine final optimized assay conditions, which are often a composite of variables that we would not have arrived at by examining each variable independently. We have applied this approach to both radioligand binding and enzymatic assays and have realized benefits in both time and performance that we would not have predicted a priori. The fully developed AAO process encompasses the ability to download information to a robot and have liquid handling methods automatically created. This evolution in smart robotics has proven to be an invaluable tool for maintaining high-quality data in the context of increasing HTS demands. 相似文献
99.
Replicon vectors derived from Sindbis virus and Semliki forest virus that establish persistent replication in host cells 总被引:1,自引:0,他引:1
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Perri S Driver DA Gardner JP Sherrill S Belli BA Dubensky TW Polo JM 《Journal of virology》2000,74(20):9802-9807
Alphavirus replicon vectors are well suited for applications where transient, high-level expression of a heterologous gene is required. Replicon vector expression in cells leads to inhibition of host macromolecular synthesis, culminating in eventual cell death by an apoptotic mechanism. For many applications, including gene expression studies in cultured cells, a longer duration of transgene expression without resulting cytopathic effects is useful. Recently, noncytopathic Sindbis virus (SIN) variants were isolated in BHK cells, and the mutations responsible were mapped to the protease domain of nonstructural protein 2 (nsP2). We report here the isolation of additional variants of both SIN and Semliki Forest virus (SFV) replicons encoding the neomycin resistance gene that can establish persistent replication in BHK cells. The SIN and SFV variant replicons resulted from previously undescribed mutations within one of three discrete regions of the nsP2 gene. Differences among the panel of variants were observed in processing of the nonstructural polyprotein and in the ratios of subgenomic to genomic RNAs. Importantly, high-level expression of a heterologous gene was retained with most replicons. Finally, in contrast to previous studies, efficient packaging was obtained with several of the variant replicons. This work expands the utility of noncytopathic replicons and the understanding of how alphavirus replicons establish persistent replication in cultured cells. 相似文献
100.