Environmental Engineering Commons^™

Performance Of Twelve Mass Transfer Based Reference Evapotranspiration Models Under Humid Climate, Koffi Djaman, Komlan Koudahe, Mamadou Sall, Isa Kabenge, Daran Rudnick, Suat Irkmak

Department of Agricultural and Biological Systems Engineering: Faculty Publications

Reference evapotranspiration is very important parameter in the hydrological, agricultural and environmental studies and is accurately estimated by the FAO Penman-Monteith equation (FAO-PM) under different climatic conditions. However, due to data requirement of the FAO-PM equation, there is a need to investigate the applicability of alternative ETo equations under limited data. The objectives of this study were to evaluate twelve mass transfer based reference evapotranspiration equations and determine the impact of ETo equation on long term water management sustainability in Tanzania and Kenya. The results showed that the Albrecht, Brockamp-Wenner, Dalto, Meyer, Rohwer and Oudin ETo equations systematically overestimated the …

Evaluation Of Valiantzas’ Simplified Forms Of The Fao-56 Penman-Monteith Reference Evapotranspiration Model In A Humid Climate, Koffi Djaman, Daran Rudnick, Valere C. Mel, Denis Mutiibwa, Lamine Diop, Mamadou Sall, Isa Kabenge, Ansoumana Bodian, Hossein Tabari, Suat Irmak

Department of Agricultural and Biological Systems Engineering: Faculty Publications

The unavailability of some meteorological variables, especially solar radiation and wind speed, is the main constraint for reference evapotranspiration (ETo) estimation using the standard United Nations Food and Agriculture Organization (FAO) Penman–Monteith (FAO-PM) equation in most developing countries. The application of ETo methods with fewer input requirements is necessary under limited climatic data conditions. The FAO-PM method under limited data conditions and nine of Valiantzas’ equations were evaluated for daily ETo estimation in a humid climate in Uganda. The FAO-PM method with missing relative humidity data performed very well across Uganda, whereas using the long-term local wind speed average values …

Crop Canopy Sensor Orientation For Late Season Nitrogen Determination In Corn, T. M. Shaver, Greg Kruger, Daran Rudnick

Department of Agricultural and Biological Systems Engineering: Faculty Publications

Increasing nitrogen use efficiency (NUE) in irrigated corn production is of great importance to overall agricultural sustainability. Studies have shown that crop canopy sensors can aid in this pursuit as they allow for the determination of nitrogen (N) requirements in split applications later in the growing season. Fertigation can also increase NUE as many split applications can be conducted. If crop canopy sensors could be used to direct N fertigation rates, overall NUE may be increased even further. However, in some cases, N differences may need to be determined later in the growing season after corn has tasseled, which can …

Spatial And Temporal Changes In Maize And Soybean Grain Yield, Precipitation Use Efficiency, And Crop Water Productivity In The U.S. Great Plains, Meetpal S. Kukal, S. Irmak

Department of Agricultural and Biological Systems Engineering: Faculty Publications

Sustainable agricultural utilization of the limited water resources demands improvements in understanding the changes in crop water productivity (CWP) in space and time, which is often presented as a potential solution to relieve the growing pressure on fresh water resources. In addition, crop yield needs to be studied in relation to precipitation received annually and during the growing season for its contribution to reduce irrigation water requirements, which is quantified through precipitation use efficiency (PUE). Hence, systematic quantifications, mapping, and analyses of large-scale CWP and PUE levels are needed. This study aims to quantify long-term (1982-2013) information on grain yield, …

Large Co2 And Ch4 Emissions From Polygonal Tundra During Spring Thaw In Northern Alaska, Naama Raz-Yaseef, Margaret S. Torn, Yuxin Wu, David P. Billesbach, Anna K. Liljedahl, Timothy J. Kneafsey, Vladimir E. Romanovsky, David R. Cook, Stan D. Wullschleger

Department of Agricultural and Biological Systems Engineering: Faculty Publications

The few prethaw observations of tundra carbon fluxes suggest that there may be large spring releases, but little is known about the scale and underlying mechanisms of this phenomenon. To address these questions, we combined ecosystem eddy flux measurements from two towers near Barrow, Alaska, with mechanistic soil-core thawing experiment. During a 2 week period prior to snowmelt in 2014, large fluxes were measured, reducing net summer uptake of CO₂ by 46% and adding 6% to cumulative CH₄ emissions. Emission pulses were linked to unique rain-on-snow events enhancing soil cracking. Controlled laboratory experiment revealed that as surface ice …

Modeling Streambank Erosion On Composite Streambanks On A Watershed Scale, A. R. Mittelstet, D. E. Storm, G. A. Fox, P. M. Allen

Department of Agricultural and Biological Systems Engineering: Faculty Publications

Streambanks can be a significant source of sediment and phosphorus to aquatic ecosystems. Although the streambank-erosion routine in the Soil and Water Assessment Tool (SWAT) has improved in recent versions, the recently developed routine in SWAT 2012 has undergone limited testing, and the lack of site or watershed specific streambank data increases the uncertainty in the streambank-erosion predictions. There were two primary objectives of this research: (1) modify and test the 2012 SWAT streambank-erosion routine on composite streambanks, and (2) compare SWAT default and field-measured channel parameters and assess their influence on predicted streambank erosion. Three modifications were made to …

Quantifying An Aquifer Nitrate Budget And Future Nitrate Discharge Using Field Data From Streambeds And Well Nests, Troy E. Gilmore, David P. Genereux, Kathleen M. Farrell, Helena Mitasova

Department of Agricultural and Biological Systems Engineering: Faculty Publications

Novel groundwater sampling (age, flux, and nitrate) carried out beneath a streambed and in wells was used to estimate (1) the current rate of change of nitrate storage, dS_NO3 /dt, in a contaminated unconfined aquifer, and (2) future [NO₃^–]_FWM (the flow-weighted mean nitrate concentration in groundwater discharge) and f_NO3 (the nitrate flux from aquifer to stream). Estimates of dS_NO3 /dt suggested that at the time of sampling (2013) the nitrate storage in the aquifer was decreasing at an annual rate (mean = –9 mmol/m²yr) equal …

Effect Of Rainfall Timing And Tillage On The Transport Of Steroidhormones In Runoff From Manure Amended Row Crop Fields, Sagor Biswas, William L. Kranz, Charles A. Shapiro, Daniel D. Snow, Shannon L. Bartelt-Hunt, Mitiku Mamo, David D. Tarkalson, Tian C. Zhang, David P. Shelton, Simon J. Van Donk, Terry L. Mader

Department of Agricultural and Biological Systems Engineering: Faculty Publications

Runoff generated from livestock manure amended row crop fields is one of the major pathways of hormone transport to the aquatic environment. The study determined the effects of manure handling, tillage methods, and rainfall timing on the occurrence and transport of steroid hormones in runoff from the row crop field. Stockpiled and composted manure from hormone treated and untreated animals were applied to test plots and subjected to two rainfall simulation events 30 days apart. During the two rainfall simulation events, detection of any steroid hormone or metabolites was identified in 8–86% of runoff samples from any tillage and manure …

Wetlands And Coastal Systems: Protecting And Restoring Valuable Ecosystems, C. T. Agouridis, K. R. Douglas-Mankin, A. C. Linhoss, A. R. Mittelstet

Department of Agricultural and Biological Systems Engineering: Faculty Publications

Wetlands and coastal systems are unique, highly productive, and often threatened landscapes that provide a host of services to both humans and the environment. This article introduces a five-article Wetlands and Coastal Systems Special Collection that evolved from a featured session at the 2015 ASABE Annual International Meeting in New Orleans, Louisiana. The Collection provides perspectives on tools and techniques for enhancing the protection and restoration of wetlands and coastal systems with emphasis on vegetation, hydrology, water quality, and planning. Topics span the Florida Everglades (two articles) and Virginia floodplain (one article) wetland systems and include remote sensing (one article) …

Unmanned Aerial Vehicles For High-Throughput Phenotyping And Agronomic Research, Yeyin Shi, J. Alex Thomasson, Seth C. Murray, N. Ace Pugh, William L. Rooney, Sanaz Shafian, Nithya Rajan, Gregory Rouze, Cristine L. S. Morgan, Haly L. Neely, Aman Rana, Muthu V. Bagavathiannan, James Henrickson, Ezekiel Bowden, John Valasek, Jeff Olsenholler, Michael P. Bishop, Ryan Sheridan, Eric B. Putman, Sorin Popescu, Travis Burks, Dale Cope, Amir Ibrahim, Billy F. Mccutchen, David D. Baltensperger, Robert V. Avant, Jr., Misty Vidrine, Chenghai Yang

Department of Agricultural and Biological Systems Engineering: Faculty Publications

Advances in automation and data science have led agriculturists to seek real-time, high quality, high-volume crop data to accelerate crop improvement through breeding and to optimize agronomic practices. Breeders have recently gained massive data-collection capability in genome sequencing of plants. Faster phenotypic trait data collection and analysis relative to genetic data leads to faster and better selections in crop improvement. Furthermore, faster and higher-resolution crop data collection leads to greater capability for scientists and growers to improve precision-agriculture practices on increasingly larger farms; e.g., site-specific application of water and nutrients. Unmanned aerial vehicles (UAVs) have recently gained traction as agricultural …

Development And Assessment Of A Groundwater Sustainability Index In Climatically Diverse Groundwater Irrigated Regions In Nebraska, Maria A. Mulet Jalil

Department of Agricultural and Biological Systems Engineering: Dissertations, Theses, and Student Research

The aim of this research was to evaluate the impact of regional change in ET on groundwater level changes and the assessment and development of a groundwater sustainability index for climatically diverse regions across Nebraska during 2000-2014. Irrigation in the selected regions is predominantly supplied by groundwater. The hypothesis is that groundwater use can become sustainable if the regional evapotranspiration (ET) is managed so that it equals the ET of vegetation that is native to the region. Site locations were Box Butte, Chase, Dundy, Holt LNNRD and York Counties and 3 ecosystems were evaluated: native vegetation, dryland and irrigated cropping …

The Experiment And Analysis Of Active Mechanisms For Enhancing Heat And Mass Transfer In Sorption Fluids, Ziqi Shen

Durham School of Architectural Engineering and Construction: Dissertations, Theses, and Student Research

This project was funded by American Society of Heating, Refrigerating, and Air-Conditioning Engineers (ASHRAE RP-1462). It is a three years’ research, including the literature review, labs construction, experiments and data analysis.

In this thesis, first of all, we conducted literature review of mechanism motion influence on heat and mass transfer and additive effect in absorption chiller. This part helps us understand the basic idea of how mechanism motion affects the heat and mass transfer of sorption fluids and gives us reference on how to select the experiment instrument and the experiment operation range.

In the second part, the instrument selection …

High-Throughput Screening Of Clinically Approved Drugs That Prime Polyethylenimine Transfection Reveals Modulation Of Mitochondria Dysfunction Response Improves Gene Transfer Efficiencies, Albert Nguyen, Jared P. Beyersdorf, Jean-Jack Riethoven, Angela K. Pannier

Department of Agricultural and Biological Systems Engineering: Faculty Publications

Nonviral gene delivery methods are advantageous over viral vectors in terms of safety, cost, and flexibility in design and application, but suffer from lower gene transfer efficiency. In addition to modifications to nucleic acid design and nonviral carriers, new tools are sought to enhance transfection. Priming is the pharmacological modulation of transfection efficiency and transgene expression, and has demonstrated transfection increase in several compounds, for example, chloroquine and glucocorticoids. To develop a library of transfection priming compounds, a highthroughput screen was performed of the NIH Clinical Collection (NCC) to identify clinical compounds that prime polyethylenimine (PEI) transfection. HEK293T cells were …

Water And Energy Use Of Antimicrobial Interventions In A Mid-Size Beef Packing Plant, Rami M. M. Ziara, Shaobin Li, Bruce I. Dvorak, Jeyamkondan Subbiah

Department of Civil and Environmental Engineering: Dissertations, Theses, and Student Research

Data regarding the water and energy usage of current antimicrobial interventions in beef packing plants is scarce. The objective of this study was to collect representative water and energy usage data in a beef packing plant, with emphasis on antimicrobial interventions, to provide baseline data for comparison of new intervention technologies developed by researchers. Permanent and portable water flow meters were installed on the plant’s plumbing system to collect water flow data from March 2014 to March 2015. A local utility company was hired to meter electricity at the different subsystems using portable data loggers. The natural gas used in …

Efficiency Of Chlorophyll In Gross Primary Productivity: A Proof Of Concept And Application In Crops, Anatoly A. Gitelson, Yi Peng, Andrés Viña, Timothy J. Arkebauer, James S. Schepers

Department of Agricultural and Biological Systems Engineering: Faculty Publications

One of the main factors affecting vegetation productivity is absorbed light, which is largely governed by chlorophyll. In this paper, we introduce the concept of chlorophyll efficiency, representing the amount of gross primary production per unit of canopy chlorophyll content (Chl) and incident PAR. We analyzed chlorophyll efficiency in two contrasting crops (soybean and maize). Given that they have different photosynthetic pathways (C3 vs. C4), leaf structures (dicot vs. monocot) and canopy architectures (a heliotrophic leaf angle distribution vs. a spherical leaf angle distribution), they cover a large spectrum of biophysical conditions. Our results show that chlorophyll efficiency in primary …

Carbon Nanotubes Affect The Toxicity Of Cuo Nanoparticles To Denitrification In Marine Sediments By Altering Cellular Internalization Of Nanoparticle, Xiong Zheng, Yinglong Su, Yinguang Chen, Rui Wan, Mu Li, Haining Huang, Xu Li

Department of Civil and Environmental Engineering: Faculty Publications

Denitrification is an important pathway for nitrate transformation in marine sediments, and this process has been observed to be negatively affected by engineered nanomaterials. However, previous studies only focused on the potential effect of a certain type of nanomaterial on microbial denitrification. Here we show that the toxicity of CuO nanoparticles (NPs) to denitrification in marine sediments is highly affected by the presence of carbon nanotubes (CNTs). It was found that the removal efficiency of total NO_X⁻-N (NO₃⁻-N and NO₂⁻-N) in the presence of CuO NPs was only 62.3%, but it …

Drinking Water: Hard Water (Calcium And Magnesium), Sharon O. Skipton, Bruce I. Dvorak

Department of Civil and Environmental Engineering: Faculty Publications

Hard water isn’t a health hazard but can be a nuisance within the home. Learn how to test water, interpret the results, and choose a treatment.

Water described as “hard” contains high amounts of dissolved calcium and magnesium. Hard water is not a health risk but can be a nuisance because of mineral buildup on plumbing fixtures and poor soap and/or detergent performance.

Water is a good solvent and picks up impurities easily. As water moves through soil and rock, it dissolves very small amounts of minerals and holds them in solution. Dissolved calcium and magnesium are the two most …

Drinking Water Treatment: Salt-Free Water “Softener” Options, Bruce I. Dvorak

Department of Civil and Environmental Engineering: Faculty Publications

New no-salt technologies may be effective in treating hard water, but consumers should carefully investigate product claims prior to buying a specific treatment device.

Hard water is not a health risk, but many Nebraska consumers find hard water to be a nuisance because of mineral buildup on plumbing fixtures, including water heaters; unsightly film on glass and crystal wares; and poor soap and/or detergent performance. Water described as “hard” contains high amounts of dissolved calcium and magnesium, which are positively charged contaminants called cations.

Water hardness often is expressed as grains of hardness per gallon of water (gpg) or milligrams …

The Influence Of Sediment Characteristics On The Fate Of Steroidogenic Compounds In Aquatic Systems And The Effects On Progesterone Bioavailability In A Target Organism, Jodi L. Sangster

Department of Civil and Environmental Engineering: Dissertations, Theses, and Student Research

There is growing concern about the biologic effects stemming from steroids in impacted waterways. In aquatic systems, interaction between steroids and sediment influence both contaminant fate as well as subsequent bioavailability to aquatic organisms. The focus of this dissertation research was to gain a better understanding of steroid behavior in aquatic systems based on the physiochemical properties of sediment and to use this knowledge to better understand the biological effects stemming from sediment-associated progesterone exposure. Two natural aquatic sediments, a sand and a silty loam, were selected to represent marked differences in sediments properties. Initially, sorption of 17β-estradiol, estrone, progesterone, …

Microbial Degradation Of Sulfonamide Antibiotics, Rachel Levine

Department of Civil and Environmental Engineering: Dissertations, Theses, and Student Research

Certain microbes can transform antibiotics in the environment. However, little is known about the identity of these microbes and their antibiotic biotransformation processes. The objectives of this study were to (1) isolate bacterial strains capable of transforming antibiotics, (2) determine the biotransformation kinetics of antibiotics, (3) characterize the effects of background carbons on the biotransformation kinetics, and (4) identify biotransformation products under various environmental conditions. Sulfadiazine (SDZ) was used as the model antibiotic in this study due to its frequent occurrence in livestock wastes. Surface soil from a cattle feedlot was collected to enrich potential SDZ degrading bacteria. A mixed …

Micro- And Nanoparticulates For Dna Vaccine Delivery, Eric Farris, Deborah M. Brown, Amanda Ramer-Tait, Angela K. Pannier

Department of Agricultural and Biological Systems Engineering: Faculty Publications

DNA vaccination has emerged as a promising alternative to traditional protein-based vaccines for the induction of protective immune responses. DNA vaccines offer several advantages over traditional vaccines, including increased stability, rapid and inexpensive production, and flexibility to produce vaccines for a wide variety of infectious diseases. However, the immunogenicity of DNA vaccines delivered as naked plasmid DNA is often weak due to degradation of the DNA by nucleases and inefficient delivery to immune cells. Therefore, biomaterial-based delivery systems based on micro- and nanoparticles that encapsulate plasmid DNA represent the most promising strategy for DNA vaccine delivery. Microparticulate delivery systems allow …

A Five-Year Performance Review Of Field-Scale, Slow-Release Permanganate Candles With Recommendations For Second-Generation Improvements, Mark Christenson, Ann Kambhu, James Reece, Steve D. Comfort, Laurie Brunner

Nebraska Water Center: Faculty Publications

In 2009, we identified a TCE plume at an abandoned landfill that was located in a low permeable siltyclay aquifer. To treat the TCE, we manufactured slow-release potassium permanganate cylinders (oxidant candles) that had diameters of either 5.1 or 7.6 cm and were 91.4 cm long. In 2010, we compared two methods of candle installation by inserting equal masses of the oxidant candles (7.6-cm vs 5.1-cm dia). The 5.1-cm dia candles were inserted with direct-push rods while the 7.6-cm candles were housed in screens and lowered into 10 permanent wells. Since installation, the 7.6-cm oxidant candles have been refurbished approximately …

Glucocorticoid Cell Priming Enhances Transfection Outcomes In Adult Human Mesenchymal Stem Cells, Abby M. Kelly, Sarah A. Plautz, Janos Zempleni, Angela K. Pannier

Department of Agricultural and Biological Systems Engineering: Faculty Publications

Human mesenchymal stem cells (hMSCs) are one of the most widely researched stem cell types with broad applications from basic research to therapeutics, the majority of which require introduction of exogenous DNA. However, safety and scalability issues hinder viral delivery, while poor efficiency hinders nonviral gene delivery, particularly to hMSCs. Here, we present the use of a pharmacologic agent (glucocorticoid) to overcome barriers to hMSC DNA transfer to enhance transfection using three common nonviral vectors. Glucocorticoid priming significantly enhances transfection in hMSCs, demonstrated by a 3-fold increase in efficiency, 4–15-fold increase in transgene expression, and prolonged transgene expression when compared …

Ecosystem Evapotranspiration: Challenges In Measurements, Estimates, And Modeling, D.M. Amatya, Suat Irmak, P. Gowda, G. Sun, J.E. Nettles, K.R. Douglas-Mankin

Department of Agricultural and Biological Systems Engineering: Faculty Publications

Evapotranspiration (ET) processes at the leaf to landscape scales in multiple land uses have important controls and feed backs for local, regional, and global climate and water resource systems. Innovative methods, tools, and technologies for improved understanding and quantification of ET and crop water use are critical for adapting more effective management strategies to cope with increasing demand for freshwater resources under global climate change. This article introduces an ASABE Special Collection of 12 articles on ET monitoring and modeling research for multiple land uses and scales. The collection focuses on recent advances in four critical topical areas: (1) reference …

Recent Morphodynamic Evolution Of The Largest Uninhibited Island In The Yangtze (Changjiang) Estuary During 1998-2014: Influence Of The Anthropogenic Interference, Wen Wei, Xuefei Mei, Zhijun Dai, Zhenghong Tang

Community and Regional Planning Program: Faculty Scholarly and Creative Activity

Estuarine geomorphology worldwide has greatly changed in the Anthropocene due to intensive human inferences in river basin and within estuary, which has received increasing global concerns. Here, recent morphodynamic evolution of Jiuduan Shoal (JDS), the largest uninhabited island in the Yangtze (Changjiang) Estuary, and associated controlling factors were analyzed based on unique high-resolution seasonal-surveyed bathymetric data during 1998–2014. It can be indicated that JDS presents novel 12 and 48 months fluctuations though significant accretion was detected on high flats above −2 m. Meanwhile, morphodynamic evolution of JDS during 1998–2014 was divided into three stages: significant siltation on land-ward half of …

2016 Nebraska Water Monitoring Programs Report, Marty Link, Ryan Chapman

Nebraska Department of Environmental Quality: Reports

The Nebraska Department of Environmental Quality (NDEQ) is charged with monitoring, assessing, and to the extent possible, managing the state’s water resources. The purpose of this work is to protect and maintain high quality water and encourage or execute activities to improve poor water quality. Monitoring is done on nearly 17,000 miles of flowing rivers and streams, more than 134,000 acres of surface water in lakes and reservoirs, as well as the vast storage of groundwater in Nebraska’s aquifers.

Long-Term Patterns Of Air Temperatures, Daily Temperature Range, Precipitation, Grass-Reference Evapotranspiration And Aridity Index In The Usa Great Plains: Part Ii. Temporal Trends, Meetpal S. Kukal, Suat Irmak

Department of Agricultural and Biological Systems Engineering: Faculty Publications

Detection of long-term changes in climate variables over large spatial scales is a very important prerequisite to the development of effective mitigation and adaption measures for the future potential climate change and for developing strategies for future hydrologic balance analyses under changing climate. Moreover, there is a need for effective approaches of providing information about these changes to decision makers, water managers and stakeholders to aid in efficient implementation of the developed strategies. This study involves computation, mapping and analyses of long-term (1968-2013) county-specific trends in annual, growing-season (1st May- 30th Sept.) and monthly air temperatures [(maximum (T_max), …

Nebraska Water Center Annual Report 2016, Nebraska Water Center

Nebraska Water Center: Administrative Materials

Contents

Foreword

Director’s Letter

Since 1964: The Nebraska Water Center

Building the future

Nebraska’s Top 10 water challenges

Nebraska Water Center advisory board

Water Resources Advisory Panel: A key to success

Nebraska Water Sciences Laboratory

USGS 104b projects for 2016

Ogallala Aquifer focus of USDA research grant

Graduate research on amphetamines draws attention

NIC hosts water symposium and law conference

Water seminar lectures key on “Water and health”

Four picked for IRES program in Czech Republic

WARI program begins

2016 tour to Colorado’s South Platte basin

45th annual tour visits Platte River basin in Colorado

McCornick new Water for Food …

Nebraska Water Center (Brochure ), Nebraska Water Center

Nebraska Water Center: Administrative Materials

The Nebraska Water Center was established in 1964 as one of 54 Water Resources Research Institutes nationwide with passage of the first Clean Water Act. It facilitates the University of Nebraska-Lincoln’s land grant missions in research, extension and teaching in water science, law and policy to address water quantity and quality issues of priority to Nebraska. NWC helps water researchers by awarding small grants, conducting water-related education and outreach activities for stakeholders, and disseminating research results through media, colloquia, conferences, lectures, and tours. NWC provides state-of-the-art analytical technology to conduct water research and education to scientists and engineers, helping them …

Protein-Targeted Corona Phase Molecular Recognition, Gili Bisker, Juyao Dong, Hoyoung D. Park, Nicole M. Iverson, Jiyoung Ahn, Justin T. Nelson, Markita P. Landry, Sebastian Kruss, Michael S. Strano

Department of Agricultural and Biological Systems Engineering: Faculty Publications

Corona phase molecular recognition (CoPhMoRe) uses a heteropolymer adsorbed onto and templated by a nanoparticle surface to recognize a specific target analyte. This method has not yet been extended to macromolecular analytes, including proteins. Herein we develop a variant of a CoPhMoRe screening procedure of single-walled carbon nanotubes (SWCNT) and use it against a panel of human blood proteins, revealing a specific corona phase that recognizes fibrinogen with high selectivity. In response to fibrinogen binding, SWCNT fluorescence decreases by >80% at saturation. Sequential binding of the three fibrinogen nodules is suggested by selective fluorescence quenching by isolated sub-domains and validated …