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Imidazolium-based ionic liquids as additives to preserve the Enhanced Green Fluorescent Protein fluorescent activity

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Green Chemical Engineering 2021年第4期2卷 412-422页

作者： Nathalia V.Veríssimo Carolina F.Saponi timothy M.Ryan Tamar l.Greaves Jorge F.B.Pereira Department of Engineering of Bioprocesses and Biotechnology School of Pharmaceutical SciencesSao Paulo State University(UNESP)Rodovia Araraquara-Jaú/Km 0114800-903AraraquaraSPBrazil School of Science College of ScienceEngineering and HealthRMIT University124 La Trobe StreetMelbourneVIC 3000Australia School of Pharmaceutical Sciences Sao Paulo University(USP)Av.Prof.Lineu Prestesno.580B1605508-000Cidade de UniversitariaSao PauloSPBrazil Australian Synchrotron 800 Blackburn RoadClaytonVIC 3168Australia Univ Coimbra CIEPQPFDepartment of Chemical EngineeringRua Sílvio LimaPolo II–Pinhal de Marrocos3030-790 CoimbraPortugal

Fluorescent Proteins(FP)can be applied as biomarkers and biosensors in the industrial and medical fields,but their large-scale use,especially for new industrial applications,is limited due to their low ***,the discovery of additives capable of preserving the activity of FP at room temperature and under stress conditions can help to expand and facilitate their commercial *** this goal,we evaluated the application of 1-alkyl-3-methylimidazolium chloride-based([C_(n)mim]Cl)ionic liquids(Ils)as additives to preserve the activity of Enhanced Green Fluorescent Protein(EGFP)at different storage times and under unfavorable ***[C_(n)mim]Cl solutions(at 0.100 mol l^(-1))were able to preserve EGFP fluorescence for longer than the phosphatesaline buffer(PBS)and NaCl solutions,increasing its fluorescence manifestation from 1 to 3 months.[C_(n)mim]Cl with shorter to medium cationic alkyl chains were the most effective in preserving EGFP fluorescence.[C_(n)mim]Cl also protected EGFP activity in the presence of the surfactant SDS,the acid guanidine hydrochloride,and H_(2)O_(2).Therefore,[C_(n)mim]Cl can be added to aqueous solutions to preserve EGFP fluorescence activity at room temperature for longer storage times and to reduce the negative impact of denaturing agents on ***,there is a massive potential for the application of Ils as additives to preserve FP in the long-term without refrigeration and under unfavorable conditions,and this is fundamental to enable expansion of FP in industrial and commercial applications.

关键词： Green Fluorescent Protein Protein stability Fluorescence Ionic liquids Preservatives

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Susceptible and Glyphosate-Resistant Palmer Amaranth (<i>Amaranthus palmeri</i>) Response to Glyphosate Using C<sup>14</sup>as a Tracer: Retention, Uptake, and Translocation

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American Journal of Plant Sciences 2018年第12期9卷 2359-2370页

作者： timothy l. Grey Donn Shilling Crop and Soil Science Department University of Georgia Athens Georgia USA

The foliar retention, absorption, translocation, and diffusion of glyphosate in glyphosate resistant-(R) and susceptible (S)-Palmer amaranth populations from seed collected in Georgia in 2007 were examined. The R population of Palmer amaranth had an elevated copy number of the EPSPS gene conferring the mechanism of resistance. When applications of 14C-glyphosate to a single leaf followed entire plant treatment with glyphosate, the distribution percentages were similar for R and S for the above and below treated leaves when harvested at 1, 6, 12, 24, and 48 hours after treatment (HAT). There were initially no differences between R and S at 1 HAT with an average of 8% absorption for both biotypes. However, data indicated that glyphosate absorption increased for R-Palmer amaranth reaching 41% within 6 HAT and was significantly different (P = 0.01) from the 28% absorbed by S-Palmer amaranth. Glyphosate resistant and susceptible Palmer amaranth averaged 44% 14C-glyphosate absorption by 24 HAT. There were no differences for 14C-glyphosate Bq/mg of plant tissue between R and S for the above the treated leaf and below the treated leaf portions of plants at 1, 6, 12, 24, or 48 HAT. However, root accumulation of 14C-glyphosate in plant tissue was significantly greater by 12 HAT for the roots of R (1.21 Bq/mg) than for S (0.51 Bq/mg). The treated leaf of the R-Palmer amaranth plants exhibited greater translocation of 14C-glyphosate in Bq/mg of tissue than the susceptible over time, indicating no detrimental effect or cost of fitness due to EPSPS gene amplification. Additionally, there were no differences in glyphosate retention in leaf discs assays between R and S biotypes. In spite of an average of 6.5 Bq efflux out of R and S leaf discs after 15 minute, only 0.4 Bq was retained after 150 minutes. Glyphosate was not retained over time in the leaf discs for R and S, and there were no biotype differences within bathing times. However, the rate of efflux (the slope of the curves

关键词： Amaranthus palmeri S. Wats Absorption Becquerel’s Glyphosate-Resistance Herbicide Resistance Translocation

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Small incision lenticule extraction (SMIlE) history, fundamentals of a new refractive surgery technique and clinical outcomes

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Eye and Vision 2014年第1期1卷 9-20页

作者： Dan Z Reinstein timothy J Archer Marine Gobbe london Vision Clinic 138 Harley StreetLondon W1G 7LAUK Department of Ophthalmology Columbia University Medical CenterNew YorkNYUSA Centre Hospitalier National d’Ophtalmologie ParisFrance

This review summarizes the current status of the small incision lenticule extraction(SMIlE)*** the early work by Sekundo et *** Shah et al.,SMIlE has become increasingly *** accuracy of the creation of the lenticule with the VisuMax femtosecond laser(Carl Zeiss Meditec)has been verified using very high-frequency(VHF)digital ultrasound and optical coherence tomography(OCT).Visual and refractive outcomes have been shown to be similar to those achieved with laser in situ keratomileusis(lASIK),notably in a large population reported by Hjortdal,Vestergaard et *** in terms of the change in corrected distance visual acuity(CDVA)has also been shown to be similar to *** was expected that there would be less postoperative dry eye after SMIlE compared to lASIK because the anterior stroma is disturbed only by the small incision,meaning that the anterior corneal nerves should be less affected.A number of studies have demonstrated a lower reduction and faster recovery of corneal sensation after SMIlE than *** studies have also used confocal microscopy to demonstrate a lower decrease in subbasal nerve fiber density after SMIlE than *** potential biomechanical advantages of SMIlE have been modeled by Reinstein et *** on the non-linearity of tensile strength through the *** have reported a similar change in Ocular Response Analyzer(Reichert)parameters after SMIlE and lASIK,however,these have previously been shown to be unreliable as a representation of corneal *** options after SMIlE are *** addition applications of the SMIlE procedure are also discussed including the potential for cryo-preservation of the lenticule for later reimplantation(Mohamed-Noriega,Angunawela,lim et al.),and a new procedure referred to as endokeratophakia in which a myopic SMIlE lenticule is implanted into a hyperopic patient(Pradhan et al.).Finally,studies reporting microdistortions in Bowman’s layer and cor

关键词： Small incision lenticule extraction(SMIlE) laser in situ keratomileusis(lASIK) lenticule Cap Dry eye Corneal sensation Corneal innervation Corneal biomechanics Endokeratophakia

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Integrated soil, water and agronomic management effects on crop productivity and selected soil properties in Western Ethiopia

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International Soil and Water Conservation Research 2018年第4期6卷 305-316页

作者： Teklu Erkossa timothy O.Williams Fanuel laekemariam International Water Management Institute P.O.Box 5689C/o ILRI Addis CampusAddis AbabaEthiopia Wolita University Ethiopia

land degradation is a major challenge limiting crop production in Ethiopia. Integrated soil and water conservation is widely applied as a means to reverse the trend and increase productivity. This study investigated the effects of such integrated approaches at two sites, Jeldu and Diga, inWestern Ethiopia. A split plot design with physical soil and water conservation in the main plots and agronomic practices in the sub plots was employed. Maize (Zea mays l.) followed by groundnut (Arachis hypogaea l.) at Diga, and wheat (Triticum aestivum) followed by faba bean (Vicia faba l.) were the test crops. Surface soils were sampled before sowing and after the crop harvest, and analyzed for selected parameters. Soil moisture content during the growing period was also monitored. The use of soil bund increased soil moisture content, and significantly (P lowering and maturity, kernel weight and harvest index, grain yield of the test crops, with the exception of maize. The improved agronomic practices (intercropping, fertilization and row planting) significantly (P ld of all the test crops. The effect of the treatments on soil parameters may require longer time to be evident. Although the increase in crop yield due to soil bund and the improved agronomic practices is eminent, economic analysis is necessary before recommending the widespread use of the improved options.

关键词： land degradation Soil bund Inter-cropping Improved crop varieties Crop-livestock systems

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Screening of Rice Cultivars for Morpho-Physiological Responses to Early-Season Soil Moisture Stress

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Rice science 2017年第6期24卷 322-335页

作者： Bhupinder SINGH Kambham Raja REDDY Edilberto Diaz REDONA timothy WAlKER Department of Plant and Soil Sciences Mississippi State UniversityMississippi 39762USA Delta Research and Extension Center Mississippi State UniversityMississippi 38776USA Horizon Ag.llC 8275 Tournament DriveSuite 255MemphisTennessee 38125USA

The majority of rice(Oryza sativa l.) produced in the southern USA is drill-seeded and grown under upland-like conditions because permanent flooding is established after the four-leaf stage. Therefore, rice during the seedling growth stage will be subjected to variable soil moisture content. A greenhouse experiment was conducted to evaluate the performance of 15 rice cultivars commonly grown in Mississippi of USA under early-season soil moisture stress. Twenty morpho-physiological parameters of rice seedlings subjected to three different levels(100%, 66% and 33% field capacity) of soil moisture, from 10 to 30 d after sowing, were measured. Significant moisture stress × treatment interaction(P l drought response index(TDRI) was developed to score the cultivars for drought tolerance with the variation from 26.88 to 36.21. Accordingly, the cultivars were classified into different groups of tolerance. The cultivars Cl152 and Cl142-AR were classified as the least and the most tolerant to drought based on TDRI and standard deviation, respectively. Even though both total root(R^2 = 0.98) or shoot(R^2 = 0.76) drought responses indices were positively correlated with TDRI, root traits were important in deriving the indices. Therefore, TDRI could be used to select cultivars for drought tolerance in a given environment and develop rice varieties with early-season drought tolerance. However, further research is needed to identify and characterize tolerance at other stages to assist breeding programs in rice.

关键词： drought field capacity morpho-physiological parameter Oryza sativa root shoot soil moisture content total drought response index

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Improving beef hamburger quality and fatty acid profiles through dietary manipulation and exploitation of fat depot heterogeneity

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Journal of Animal Science and Biotechnology 2015年第1期6卷 111-121页

作者： Cletos Mapiye Jennifer l Aalhus Payam Vahmani David C Rolland timothy A McAllister Hushton C Block Bethany Uttaro Spencer D Proctor Michael E R Dugan Agriculture and Agri-Food Canada Lacombe Research Centre6000 C and E TrailLacombeAlberta T4L 1 W1Canada Department of Animal Sciences Faculty of AgriSciencesStellenbosch UniversityP.Bag X1Matieland 7602 South Africa Agriculture and Agri-Food Canada Lethbridge Research Centre1st Avenue South 5403PO Box 3000LethbridgeAlberta T1J 4B1 Canada Metabolic and Cardiovascular Diseases laboratory Diabetes and Mazankowski InstitutesLi Ka Shing Centre for Health Research Innovation University of AlbertaEdmontonAlberta T6G 2E1Canada

Background： Hamburger is the most consumed beef product in North America, but lacks in nutritional appeal due to its high fat content and high proportion of saturated fatty acids （SFA）. Objectives of the present study were to improve the FA profiles of hamburgers made with perirenal fat （PRF） and subcutaneous fat （SCF） when feeding steers different diets along with examining differences in sensory attributes and oxidative stability. Diets included a control diet containing 70：30 red clover silage： barley based concentrate, a diet containing sunflower-seed （SS） substituted for barley, and diets containing SS with 15% wheat dried distillers＇ grain with solubles （DDGS-15） or 30% DDGS （DDGS-30）. Hamburgers were made from triceps brachii and either PRF or SCF （80：20 w/w）. Results： Perirenal fat versus SCF hamburgers FA had 14.3% more （P 〈0.05） 18：0, 11.8% less cis （c）9-18：1 （P 〈0.05）, and 1.82% more total trdns （t）-18：1 mainly in the form of tl 1-18：1. During sensory evaluation, PRF versus SCF hamburgers had greater （P 〈0.05） mouth coating, but the difference was less than one panel unit. Examining effects of steer diet within PRF hamburgers, feeding the SS compared to the control diet increased （P 〈0.05） t-18：1 by 2.89% mainly in the form of tl 1-18：1, feeding DGGS-15 diet led to no further changes （P 〉0.05）, but feeding DDGS-30 diet reduced the proportions of （P 〈0.05） of t-18：1 chiefly tl 1-18：1. Feeding SS and DDGS diets had small but significant （P 〈0.05） effects on hamburger sensory attributes and oxidative stability. Conclusions： Feeding high-forage diets including SS and 15% DDGS, and taking advantage of the FA heterogeneity between fat depots offers an opportunity to differentially enhance beef hamburgers with 18：2n-6 biohydrogenation products （le., t11-18：1） with potential human health benefits without compromising their sensory attributes and oxidative stability during retail display.

关键词： Beef DDGS Fat depot Fatty acids Sensory attributes Oxidative stability

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Driver mutations of cancer epigenomes

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Protein & Cell 2014年第4期5卷 265-296页

作者： David M. Roy logan A. Walsh timothy A. Chan Human Oncology and Pathogenesis Program Memorial Sloan-Kettering Cancer Center New York NY 10065 USA Weill Cornell/Rockefeller/Sloan-Kettering Tri-lnstitutional MD-PhD Program New York NY 10065 USA Department of Radiation Oncology Memorial Sloan-Kettering Cancer Center New York NY 10065 USA

Epigenetic alterations are associated with all aspects of cancer, from tumor initiation to cancer progression and metastasis. It is now well understood that both losses and gains of DNA methylation as well as altered chro- matin organization contribute significantly to cancer- associated phenotypes. More recently, new sequencing technologies have allowed the identification of driver mutations in epigenetic regulators, providing a mecha- nistic link between the cancer epigenome and genetic alterations. Oncogenic activating mutations are now known to occur in a number of epigenetic modifiers （i.e. IDHll2, EZH2, DNMT3A）, pinpointing epigenetic path- ways that are involved in tumorigenesis. Similarly, investigations into the role of inactivating mutations in chromatin modifiers （i.e. KDM6A, CREBBPIEP300, SMARCB1） implicate many of these genes as tumor suppressors. Intriguingly, a number of neoplasms are defined by a plethora of mutations in epigenetic regu- lators, including renal, bladder, and adenoid cystic car- cinomas. Particularly striking is the discovery of frequent histone H3.3 mutations in pediatric glioma, a particularly aggressive neoplasm that has long remained poorly understood. Cancer epigenetics is a relatively new, promising frontier with much potential for improving cancer outcomes. Already, therapies such as 5-azacytidine and decitabine have proven that targeting epigenetic alterations in cancer can lead to tangible benefits. Understanding how genetic alterations give rise to the cancer epigenome will offer new possibilities for developing better prognostic and therapeutic strategies.

关键词： chromatin cancer epigenetics mutations,methylation

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Improving the electrochemical performance of α-MoO_(3) electrode using aluminium trifluoromethanesulfonate water-in-salt electrolyte

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Journal of Energy Chemistry 2023年第3期78卷 123-134,I0004页

作者： Ayman E.Elkholy timothy T.Duignan Ruth Knibbe Xiu Song Zhao School of Chemical Engineering The University of QueenslandSt Lucia CampusBrisbane 4072Australia School of Mechanical and Mining Engineering The University of QueenslandSt Lucia CampusBrisbane 4072Australia

Orthorhombic molybdenum trioxide(α-MoO_(3)) electrode material experiences severe capacity fading and poor cycling stability in aqueous *** investigated the charge-storage performance of α-MoO_(3) electrode in aluminium trifluoromethanesulfonate(Al(OTf)_(3))-based salt-in-water electrolyte(SiWE) and water-in-salt electrolyte(WiSE).It was found that α-MoO_(3) electrode exhibits significantly different cycling stabilities in both electrolytes with capacity retentions of 8% using the former and87% using the *** is because α-MoO_(3) electrode maintains its crystal structure upon cycling in WiSE,but experiences substantial structural collapses and partial dissolution upon cycling in *** behaviour was inferred from both operando electrogravimetry and ex situ *** results suggest that the predominant charge-storage mechanism in a-MoO_(3) electrode using WiSE is the intercalation of protons produced from electrolyte hydrolysis with some contribution from surface pseudocapacitance enabled by Al3+ions.A two-volt full cell fabricated from α-MoO_(3) electrode as anode and copper hexacyanoferrate(CuHCF) electrode as cathode using WiSE delivers volumetric and gravimetric energies of 10.4 Wh/l and 26.5 Wh/kg,respectively,with 78% capacity retention after 2500 *** study provides an insightful understanding of the electrochemical performance of α-MoO_(3) electrode in Al(OTf)_(3)-based electrolytes.

关键词： Molybdenum oxide Aqueous electrolyte Water-in-salt electrolyte Aqueous batteries Pseudocapacitance

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Early microrecanalization of vas deferens following biodegradable graft implantation in bilaterally vasectomized rats

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Asian Journal of Andrology 2009年第3期11卷 373-378页

作者： Christopher M. Simons Barry R. De Young Thomas S. Griffith timothy l. Ratliff Erin Jones Surya K. Mallapragada Moshe Wald Department of Urology University of Iowa Iowa City IA 52242-1089 USA Department of Pathology University of Iowa Iowa City IA 52242-1089 USA Department of Chemical and Biological Engineering Iowa State University Ames IA USA

We evaluated a biodegradable graft for reconstruction of rat vasa deferentia with long obstructed or missing segments. A total of 47 Sprague-Dawley rats underwent bilateral vasectomy and were divided into groups according to length of the vas deferens affected （0.5, 1, 1.5 cm）. After 8 weeks, poly-（D,l-lactide）（PDlA） grafts were used to reconnect the vas deferens, Grafts and adjoining vasa deferentia were excised 8 and 12 weeks later and evaluated microscopically. At 8 weeks, microscopic changes included a robust inflammatory response around the grafts. All grafts were still intact but in the early stages of degradation. No microtubules, indicative of vas deferens recanalization, were identified. One specimen showed evidence of healing and neovascularization at the interface zone between the vas deferens and the graft. At 12 weeks, grafts were further degraded but still present. Microscopic evaluation showed decreased inflammation. Seven specimens showed neovascularization at the interface zone; two of these showed distinct epithelialized vas deferens microcanals at the graft edges. One specimen showed a microcanal spanning the entire 0.5-cm graft. A time period of 8 weeks is not ample enough for vas deferens regeneration in the setting of a biodegradable PDlA graft; however, early evidence of re-growth was seen at 12 weeks. A longer healing time should permit further biodegradation of the graft, as well as re-growth and possible eventual reconnection of the vas deferens, allowing passage of sperm. These findings suggest a potential role for biodegradable grafts in the reconstruction ofvas deferens with long obstructed segments.

关键词： graft infertility male vas deferens

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Early onset of a microcystin-producing cyanobacterial bloom in an agriculturally-influenced Great lakes tributary

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Journal of Oceanology and limnology 2018年第4期36卷 1112-1125页

作者： Robert Michael l.McKAY Taylor TUTTlE laura A.REITZ George S.BUllERJAHN William R.CODY Adam J.McDOWEll timothy W.DAVIS Department of Biological Sciences Bowling Green State University Bowling Green OH 43403 USA Aquatic Taxonomy Specialists Malinta OH 43535 USA City of Defiance Water Division Defiance OH 43512 USA NOAA Great lakes Environmental Research laboratory Ann Arbor MI USA

In late May 2016, a cyanobacterial harmful algal bloom(cHAB) was detected in the Maumee River, the largest tributary to lake Erie, the southernmost lake of the laurentian Great lakes system. Testing on 31 May identified Planktothrix agardhii as the dominant cyanobacterium with cell abundance exceeding 1.7×10~9 cells/l and total microcystins(MC) reaching 19 μg/l MC-lR equivalents, a level over 10-fold higher than the 2015 revised U.S. Environmental Protection Agency(EPA) national health advisory levels for drinking water exposure to adults. low river discharge coincident with negligible precipitation through the latter half of May coincided with an 80% decline in river turbidity that likely favored bloom formation by a low-light adapted P. agardhii population. Also contributing to the c HAB were high initial nutrient loads and an increase of the river temperature from 13℃ to 26℃ over this same period. The bloom persisted through 5 June with microcystins exceeding 22 μg/l MC-lR equivalents at the bloom peak. By 6 June, the river had returned to its muddy character following a rain event and sampling on 7 June detected only low levels of toxin(l) at public water systems located near the bloom origin. The elevated toxin production associated with this early onset bloom was without precedent for the Maumee River and an unique attribute of the c HAB was the high proportion of potentially-toxic genotypes. Whereas Planktothrix spp. is common in lotic environments, and has been previously detected in the Maumee, blooms are not commonly reported. This early onset, microcystin-producing c HAB provided a rare opportunity to glean insights into environmental factors that promote bloom development and dominance by Planktothrix in lotic environments.

关键词： cyanobacteria microcystin Planktothrix qPCR river

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