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Generation of a human embryonic stem cell line stably expressing high levels of the fluorescent protein mCherry

World Journal of Stem Cells 2012年第7期4卷 71-79页

作者： Dmitry A Ovchinnikov Jennifer P Turner drew m titmarsh Nilay Y Thakar Dong Choon Sin Justin J Cooper-White Ernst J Wolvetang Stem Cell Engineering Group Australian Institute for Bioengineering and NanotechnologyUniversity of QueenslandBrisbane 4072QueenslandAustralia Tissue Engineering and microfluidics Laboratory Australian Institute for Bioengineering and NanotechnologyUniversity of QueenslandBrisbane 4072QueenslandAustralia School of Chemical Engineering University of Queensland Brisbane 4072QueenslandAustralia

AIm:The generation and characterization of a human embryonic stem cell (hESC) line stably expressing red fluorescent mCherry protein. mETHODS:Lentiviral transduction of a ubiquitously-expressed human EF-1α promoter driven mCherry transgene was performed in mEL2 hESC. Red fluore-scence was assessed by immunofluorescence and flow cytometry. Pluripotency of stably transduced hESC was determined by immunofluorescent pluripotency marker expression, flow cytometry, teratoma assays andembryoid body-based differentiation followed by reverse transcriptase-polymerase chain reaction. Quantification of cell motility and survival was performed with time lapse microscopy. RESULTS:Constitutively fluorescently-labeled hESCs are useful tools for facile in vitro and in vivo tracking of survival, motility and cell spreading on various surfaces before and after differentiation. Here we describe the generation and characterization of a hESC line (mEL2) stably expressing red fluorescent protein, mCherry. This line was generated by random integration of a fluorescent protein-expressing cassette, driven by the ubiquitously-expressed human EF-1α promoter. Stably transfected mEL2-mCherry hESC were shown to express pluripo-tency markers in the nucleus (POU5F1/OCT4, NANOG and SOX2) and on the cell surface (SSEA4, TRA1-60 and TG30/CD9) and were shown to maintain a normal karyotype in long-term (for at least 35 passages) culture. mEL2-mCherry hESC further readily differentiated into representative cell types of the three germ layers in embryoid body and teratoma based assays and, importantly, maintained robust mCherry expression throughout differentiation. The cell line was next adapted to singlecell passaging, rendering it compatible with numerous bioengineering applications such as measurement of cell motility and cell spreading on various protein modified surfaces, quantification of cell attachment to nanoparticles and rapid estimation of cell survival. CONCLUSION:The mEL2-mCherry hESC line c

关键词： Human embryonic stem cells Fluorescent marker mCherry Pluripotency Cellular motility

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Growth at the microscale:long term thinning effects on patterns and timing of intra-annual stem increment in radiata pine

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Forest Ecosystems 2018年第4期5卷 438-450页

作者： David m.drew Geoffrey m.Downes Department of Forest and Wood Science Stellenbosch UniversityPrivate Bag X1Matieland 7800South Africa Forest Quality Pty Ltd PO Box 293 HuonvilleTASAustralia.

Background: Stem radial growth in forests is not uniform. Rather, it is characterized by periods of relatively fast or slow growth, or sometimes no growth at all. These fluctuations are generally a function of varying environmental conditions(e.g. water availability) and, importantly, will also be associated with adjustments in proprties in the wood formed. Stand level conditions and forest management, particularly thinning and stand density will, thowever, also have a major influence on patterns of growth variation. We explore how different thinning histories and/or stand densities influence these dynamics of tree growth in the important commercial plantation species Pinus radiata D. ***: Daily stem size change was measured using electronic point dendrometers over two growing seasons on P. radiata trees at two sites, subjected to different thinning regimes. Timing, rates and periodicity of annual growth were calculated from these ***: Greater overall cross-sectional growth in thinned plots was driven mainly by two dynamics. First, the cessation of seasonal growth occurred at least 3 weeks later in the stands in which thinning had taken place. There was no difference between thinned/unthinned stands, however, in the timing of growth onset. Second, within the longer season, trees in thinned plots had more growth days(as much as 20% more) than unthinned plots. The rates of growth on days when growth occurred were not different, however. In this context, it is notable that in trees in the unthinned plots experiencing the most severe competition there were strong "pulses" of growth following drought-breaking rainfall events. Unthinned plots at high stand densities also maintained a smaller(but consistent) zone of dividing cells throughout the season than thinned ***: In Pinus radiata growing under conditions as in our study, conditions late in summer, particularly drought, have an important effect on the timing of cessation of growth. Earl

关键词： monterey pine Dendrometer Intra-annual growth Drought Thinning

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Who＇s your neighbor？ Acoustic cues to individual identity in red squirrel Tamiasciurus hudsonicus rattle calls

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Current Zoology 2012年第5期58卷 758-764页

作者： Shannon m. DIGWEED drew RENDALL Teana ImBEAU Departments of Psychology and Biological Science Grant MacEwan University Edmonton Alberta T5J 2P2 Canada Department of Psychology University of Lethbridge Lethbridge Alberta T1K 3M4 Canada Department of Biological Science Grant MacEwan University Edmonton Alberta T5J 2P2 Canada

North American red squirrels Tamiasciurus hudsonicus often produce a loud territorial rattle call when conspecifics enter or invade a territory. Previous playback experiments suggest that the territorial rattle call may indicate an invader＇s identity as squirrels responded more intensely to calls played from strangers than to calls played from neighbors. This dear-enemy effect is well known in a variety of bird and mammal species and functions to reduce aggressive interactions between known neighbors. However, although previous experiments on red squirrels suggest some form of individual differentiation and thus recognition, detailed acoustic analysis of potential acoustic cues in rattle calls have not been conducted. If calls function to aid in conspecific identifi- cation in order to mitigate aggressive territorial interactions, we would expect that individual recognition cues would be acoustically represented. Our work provides a detailed analysis of acoustic cues to identity within rattle calls. A total of 225 calls across 32 in- dividual squirrels from Sheep River Provincial Park, Kananaskis, AB, Canada, were analyzed with discriminant function analysis for potential acoustic cues to individual identity. Initial analysis of all individuals revealed a reliable acoustic differentiation across individuals. A more detailed analysis of clusters of neighboring squirrels was performed and results again indicated a statistically significant likelihood that calls were assigned correctly to specific squirrels （55%-75% correctly assigned）; in other words squirrels have distinct voices that should allow for individual identification and discrimination by conspecifics [Current Zoology 58 （5）： 758-764, 2012].

关键词： Vocal communication Territory calls Red squirrel Dear-enemy Individual identity

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Biomechanics associated with tibial stress fracture in runners:A systematic review and meta-analysis

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Journal of Sport and Health Science 2023年第3期12卷 333-342页

作者： Clare E.milner Eric Foch Joseph m.Gonzales drew Petersen Department of Physical Therapy and Rehabilitation Sciences Drexel UniversityPhiladelphiaPA 19102USA Department of Health Sciences Central Washington UniversityEllensburgWA 98926USA

Background:Tibial stress fracture(TSF)is an overuse running injury with a long recovery *** many running studies refer to biomechanical risk factors for TSF,only a few have compared biomechanics in runners with TSF to *** aim of this systematic review and meta-analysis was to evaluate biomechanics in runners with TSF compared to ***:Electronic databases Pubmed,Web of Science,SPORTDiscus,Scopus,Cochrane,and CINAHL were *** of bias was assessed and meta-analysis conducted for variables reported in 3 or more ***:The search retrieved 359 unique records,but only the 14 that compared runners with TSF to controls were included in the *** studies were retrospective,2 were prospective,and most had a small sample size(5-30 per group).many variables were not significantly different between ***-analysis of peak impact,active,and braking ground reaction forces found no significant differences between *** studies found larger tibial peak anterior tensile stress,peak posterior compressive stress,peak axial acceleration,peak rearfoot eversion,and hip adduction in the TSF ***:meta-analysis indicated that discrete ground reaction force variables were not statistically significantly different in runners with TSF compared to *** individual included studies,many biomechanical variables were not statistically significantly different between ***,many were reported by only a single study,and sample sizes were *** encourage additional studies with larger sample sizes of runners with TSF and controls and adequate statistical power to confirm or refute these findings.

关键词： Bone stress injury Gait Kinematics Kinetics Tibial acceleration

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Particle Size Optimization of Thermochemical Salt Hydrates for High Energy Density Thermal Storage

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Energy & Environmental materials 2024年第2期7卷 326-333页

作者： Andrew martin drew Lilley Raνi Prasher Sumanjeet Kaur Energy Technologies Area Lawrence Berkeley National LaboratoryBerkeleyCA 94720USA Department of mechanical Engineering UC BerkeleyBerkeleyCA 94720USA

Thermal energy storage(TES)solutions offer opportunities to reduce energy consumption,greenhouse gas emissions,and ***,they can help reduce the peak load and address the intermittency of renewable energy sources by time shifting the load,which are critical toward zero energy *** materials(TCms)as a class of TES undergo a solid-gas reversible chemical reaction with water vapor to store and release energy with high storage capacities(600 kWh m^(-3))and negligible self-discharge that makes them uniquely suited as compact,stand-alone units for daily or seasonal ***,TCms suffer from instabilities at the material(salt particles)and reactor level(packed beds of salt),resulting in poor multi-cycle efficiency and high-levelized cost of *** this study,a model is developed to predict the pulverization limit or Rcrit of various salt hydrates during thermal *** is critical as it provides design rules to make mechanically stable TCm composites as well as enables the use of more energy-efficient manufacturing process(solid-state mixing)to make the *** model is experimentally validated on multiple TCm salt hydrates with different water content,and effect of Rcrit on hydration and dehydration kinetics is also investigated.

关键词： high energy density hydration kinetics long-term cycling thermal energy storage thermochemical materials

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Integrative filtration research and sustainable nanotechnology

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Particuology 2013年第1期11卷 5-13页

作者： Jing Wang drew Thompson David Y.H.Pui Institute of Environmental Engineering ETH Zurich 8093Switzerland Analytical Chemistry Empa8600 DubendorfSwitzerland Particle Technology Laboratory University of Minnesota55455USA

With the wide applications of nanomaterials in an array of industries, more concerns are being raised about the occupational health and safety of nanoparticles in the workplace, and implications ofnanotech- nology on the environment and living systems. Studies on environmental, health and safety （EHS） issues of nanomaterials play a significant role in public acceptance, and eventual sustainability, of nanotechnol- ogy. We present research results on three aspects of the EHS studies： characterization and measurement of nanoparticles, nanoparticle emission and exposure at workplaces, and control and abatement of nanoparticle release using filtration technology. measurement of nanoparticle agglomerates using a newly developed instrument, the Universal Nanoparticle Analyzer, is discussed. Nanoparticle emission and exposure measurement results for carbon nanotubes in the manufacture of nanocomposites and for silicon nanoparticles in their production at a pilot scale facility are presented. Filtration of nanoparticles and nanoparticle agglomerates are also studied.

关键词： Agglomerate Sustainability Exposure Filtration Nano-EHS Nanoparticle

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Life cycle assessment of low impact development technologies combined with conventional centralized water systems for the City of Atlanta, Georgia

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Frontiers of Environmental Science & Engineering 2016年第6期10卷 3-15页

作者： Hyunju Jeong Osvaldo A. Broesicke Bob drew Duo Li John C. Crittenden Department of Civil and Environmental Engineering South Dakota School of Mines and Technology Rapid City SD 57701 USA Brook Byers Institute for Sustainable Systems School of Civil and Environmental Engineering Georgia Institute of Technology 828 West Peachtree Street Suite 320 Atlanta GA 30332-0595 USA ECOVIE Rainwater Collection Systems 4287 Club Drive N.E. Atlanta GA 30319 USA Crittenden and Associates C-305 Building E Wangjing High-tech Park LizezhongEr Road Chaoyang District Beijing 100102 China

Low-impact development （LID） technologies, such as bioretention areas, rooftop rainwater harvesting, a_nd xeris_caping can co_ntrol stormwater runoff, supply non-potable water, and landscape open *** study examines a hybrid system （HS） that combines LID technologies with a centralized water system to lessen the burden on a conventional system （CS）. CS is defined as the stormwater collection and water supply infrastructure, and the conventional landscaping choices in the City of Atlanta. The study scope is limited to five single-family residential zones （SFZs）, classified R-1 through R-5, and four multi-family residential zones （mFZs）, classified RG-2 through RG-5. Population density increases from 0.4 （R-1） to 62.2 （RG-5） persons per 1,000 m2. We performed a life cycle assessment （LCA） comparison of CS and HS using TRACI 2.1 to simulate impacts on the ecosystem, human health, and natural resources. We quantified the impact of freshwater consumption using the freshwater ecosystem impact （FEI） indicator. Test results indicate that HS has a higher LCA single score than CS in zones with a low population density; however, the difference becomes negligible as population density increases. Incorporating LID in SFZs and mFZs can reduce potable water use by an average of 50%. and 25%,respectively.; however, water savings are negligible in zones with high population density （i.e., RG-5） due to the diminished surface area per capitaavailable for LID technoogies. The results demonstrate that LID technologies effectively reduce outdoor water demand and therefore would be a good choice to decrease the water consumption impact in the City of Atlanta.

关键词： Life cycle assessment (LCA) Low impact development (LID) Bioretention area Rainwater harvesting Xeriscaping

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Diversity and production in an Afromontane Forest

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Forest Ecosystems 2016年第4期3卷 275-286页

作者： Klaus v.Gadow GongQiao Zhang Graham Durrheim David drew Armin Seydack Georg-August University in Gottingen Gottingen Germany Department of Forestry and Wood Science University of Stellenbosch Stellenbosch South Africa Chinese Academy of Forestry in Beijing Beijing China South African National Parks Knysna South Africa

Background：This contribution evaluates the effect of forest structure and tree species diversity on plot productivity and individual tree growth in the unique Knysna forests in Southern Africa using mapped tree data from an observational study that has been re-measured over a period of 40 ***：The effects of tree species diversity and forest structure on tree growth and forest production are evaluated on three levels of resolution：a） the forest community（canopy,sub-canopy species）,b） the subplots（number of trees per ha,skewness of the diameter distribution,diameter coefficient of variation） and c） the immediate neighborhood of selected reference trees（＂mingling＂,＂Dominance＂,Aggregation＂ and ＂Size Variation＂）.Results：An analysis of the community level identified two distinct clusters,one including dominant/canopy species with the highest growth rates and a greater variation of growth,and another cluster which includes the remaining subcanopy species which have a smaller maximum size and lower rates of *** area-based structure variables on plot level have a highly significant effect on total basal area ***,the effects of forest density and species richness on productivity were not straight *** basal area production of about 0.75 m^2/ha/year is achieved at medium levels of richness（around 20 species per ha） and medium levels of density（around 30 m^2/ha basal area） using percentile regression *** relative ＂Dominance＂ of a selected reference tree had a highly significant effect on individual tree growth on all investigated *** neighbourhood structure variables were only occasionally significant or not significant at ***：This contribution presents a new theoretical framework for analysing natural forests that includes community,plot and neighborhood variables of forest structure and diversity,and a first specific analysis of the structure and dynamics of the Knysna Afromontane Forest,based on a unique s

关键词： canopy neighborhood forests richness basal straight productivity occasionally remaining tropical

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Systematic development of ionizable lipid nanoparticles for placental mRNA delivery using a design of experiments approach

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Bioactive materials 2024年第4期34卷 125-137页

作者： Rachel E.Young Katherine m.Nelson Samuel I.Hofbauer Tara Vijayakumar mohamad-Gabriel Alameh drew Weissman Charalampos Papachristou Jason P.Gleghorn Rachel S.Riley Department of Biomedical Engineering Henry M.Rowan College of EngineeringRowan University201 Mullica Hill RdGlassboroNJ 08028United States School of Translational Biomedical Engineering&Sciences Virtua College of Medicine&Life Sciences of Rowan University201 Mullica Hill RdGlassboroNJ 08028United States Department of Chemical and Biomolecular Engineering College of EngineeringUniversity of Delaware150 Academy StreetNewarkDE 19716United States Cooper medical School of Rowan University Rowan University401 BroadwayCamdenNJ 08103United States Perelman School of medicine University of Pennsylvania3400 Civic Center BlvdPhiladelphiaPA 19104United States Department of mathematics College of Science&MathematicsRowan University201 Mullica Hill RdGlassboroNJ 08028United States Department of Biomedical Engineering College of EngineeringUniversity of Delaware590 Avenue 1743NewarkDE 19713United States

Ionizable lipid nanoparticles(LNPs)have gained attention as mRNA delivery platforms for vaccination against COVID-19 and for protein replacement *** enhance mRNA stability,circulation time,cellular uptake,and preferential delivery to specific tissues compared to mRNA with no carrier ***,LNPs are only in the beginning stages of development for safe and effective mRNA delivery to the placenta to treat placental ***,we develop LNPs that enable high levels of mRNA delivery to trophoblasts in vitro and to the placenta in vivo with no *** conducted a Design of Experiments to explore how LNP composition,including the type and molar ratio of each lipid component,drives trophoblast and placental *** data revealed that utilizing C12-200 as the ionizable lipid and 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine(DOPE)as the phospholipid in the LNP design yields high transfection efficiency in *** of lipid molar composition as a design parameter in LNPs displayed a strong correlation between apparent pKa and poly(ethylene)glycol(PEG)content,as a reduction in PEG molar amount increases apparent ***,we present one LNP platform that exhibits the highest delivery of placental growth factor mRNA to the placenta in pregnant mice,resulting in synthesis and secretion of a potentially therapeutic ***,our high-performing LNPs have no toxicity to both the pregnant mice and *** results demonstrate the feasibility of LNPs as a platform for mRNA delivery to the placenta,and our top LNP formulations may provide a therapeutic platform to treat diseases that originate from placental dysfunction during pregnancy.

关键词： Lipid nanoparticles(LNPs) Drug delivery Nucleic acids Placental growth factor(PlGF) Placenta Pregnancy Preeclampsia

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Effects of Aggregate Extraction on Water Storage in the Pepin Creek Watershed, British Columbia, Canada

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Natural Resources 2017年第7期8卷 461-477页

作者： Yining Wang Julie E. Wilson drew Brayshaw Les m. Lavkulich master of Land and Water Systems Faculty of Land and Food Systems The University of British Columbia Vancouver Canada Statlu Environmental Consulting Ltd. Chilliwack Canada

Aggregate extraction in the Lower Fraser Valley of British Columbia, Canada, has affected the soil water storage of the Pepin Creek watershed. Although local government has set regulations for aggregate extraction projects to avoid negative environmental impacts, the gradual loss of soil materials and associated changes in vegetative cover has led to an alteration of the water balance within the watershed, which may affect surface or groundwater levels, and aquatic habitats. The study assessed the effects of aggregate extraction on the water storage of the Pepin Creek watershed and estimated that 25% of the surface area of the Canadian portion of the watershed has been affected by aggregate mining with an estimated loss of water storage of 10%. Evapotranspiration has decreased as a result of the removal of the vegetative cover. Precipitation has remained relatively constant over the study period but the annual discharge measured at Pepin Creek has decreased. Recommendations for enhancing environmental monitoring to better measure and understand ecological functions of the watershed during aggregate extraction are provided.

关键词： Soil Water Storage Aggregate Extraction Water Balance Watershed Storage

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