Development of Elispot and CFSE flow cytometric assays for detecting beryllium sensitivity

Background: Beryllium sensitization (BeS) is caused by exposure to beryllium in the workplace and may progress to chronic beryllium disease (CBD). This granulomatous lung disorder mimicks sarcoidosis clinically, but is characterized by beryllium specific CD4+ T-cells immune response. BeS is classically detected by beryllium lymphocyte proliferation test (BeLPT), but this assay requires radioactivity and is not very sensitive. In the context of a study aiming to evaluate if CBD patients are misdiagnosed as sarcoidosis patients in Switzerland, we developed EliSpot and CFSE beryllium flow cytometric test. Methods: 23 patients considered as having sarcoidosis (n = 21), CBD (n = 1) and possible CBD (n = 1) were enrolled. Elispot was performed using plate covered with gamma-IFN mAb. Cells were added to wells and incubated overnight at 37 °C with medium (neg ctrl), SEB (pos ctrl) or BeSO4 at 1, 10 and 100 microM. Anti-IFN-gamma biotinylated mAb were added and spots were visualized using streptavidinhorseradish peroxidase and AEC substrate reagent. Results were reported as spot forming unit (SFU). For Beryllium specific CFSE flow cytometry analysis, CFSE labelled cells were cultured in the presence of SEB and 1, 10 or 100 microM BeSO4. Unstimulated CFSE labeled cells were defined as controls. The cells were incubated for 6 days at 37 °C and 5% CO2. Surface labelling of T-lymphocytes and vivid as control of cells viability was performed at the time of harvest. Results: Using EliSpot technology, we were able to detect a BeS in 1/23 enrolled patients with a mean of 780 SFU (cut off value at 50 SFU). This positive result was confirmed using different concentration of BeSO4. Among the 23 patients tested, 22 showed negative results with EliSpot. Using CFSE flow cytometry, 1/7 tested patients showed a positive result with a beryllium specific CD4+ count around 30% versus 45% for SEB stimulation as positif control and 0.6 % for negative control. This patient was the one with a positive EliSpot assay. Conclusions: The preliminary data demonstrated the feasibility of Elispot and CFSE flow cytometry to detect BeS. The patient with a beryllium specific positive EliSpot and CFSE flow cytometry result had been exposed to beryllium at her workplace 20 years ago and is still regularly controlled for her pulmonary status. A positive BeLPT had already been described in 2001 in France for this patient. Further validation of these techniques are in progress.

Development of a Rational Characterization Method for Iowa Fly Ash, HR-286, 1988

This research project was conducted in an attempt to determine the cause of paste strength variability in Iowa fly ashes and to develop test methods to more adequately reflect fly ash physical and chemical characteristics. An extensive three year sampling and testing program was developed and initiated which incorporated fly ash from several Iowa power plants. Power plant design and operating data were collected. The variability was directly linked to power plant maintenance schedules and to sodium carbonate coal pretreatment. Fly ash physical and chemical properties can change drastically immediately before and after a maintenance outage. The concentrations of sulfate bearing minerals in the fly ash increases sharply during shutdown. Chemical, mineralogical, and physical testing indicated that the sodium, sulfate bearing minerals, lime and tricalcium aluminate contents of the fly ashes play important roles in the development of hydration reaction products in fly ash pastes. The weak pastes always contained ettringite as the major reaction product. The strong pastes contained straetlingite and monosulfoaluminate as the major reaction products along with minor amounts of ettringite. Recommendations for testing procedure changes and suggested interim test methods are presented.

Iowa Development of Roller Compacted Concrete, HR-300, 1989

Roller compacted concrete (RCC) is a zero slump portland cement concrete mixture that has been used since the early 1970's in massive concrete structures. Iowa Highway Research Board project HR-300 was established to determine if this type mix could be used to pave roads on the Iowa road system. Manatt's Inc. of Brooklyn, Iowa agreed to pave an 800 ft. x 22 ft. x 10 in. section of RCC pavement in their Ames construction yard. This report discusses the construction of the test slab and interprets test results conducted during and after construction. It was observed that RCC can be placed with conventional asphalt paving equipment. However, there are several problems with RCC paving which must be resolved before RCC can become a viable paving alternative on Iowa's roadway system.

Development of an Expert System for Forecasting Frost on Bridges and Roadways in Iowa, HR-305, 1991

An expert system has been developed that provides 24 hour forecasts of roadway and bridge frost for locations in Iowa. The system is based on analysis of frost observations taken by highway maintenance personnel, analysis of conditions leading to frost as obtained from meteorologists with experience in forecasting bridge and roadway frost, and from fundamental physical principles of frost processes. The expert system requires the forecaster to enter information on recent maximum and minimum temperatures and forecasts of maximum and minimum air temperatures, dew point temperatures, precipitation, cloudiness, and wind speed. The system has been used operationally for the last two frost seasons by Freese-Notis Associates, who have been under contract with the Iowa DOT to supply frost forecasts. The operational meteorologists give the system their strong endorsement. They always consult the system before making a frost forecast unless conditions clearly indicate frost is not likely. In operational use, the system is run several times with different input values to test the sensitivity of frost formation on a particular day to various meteorological parameters. The users comment. that the system helps them to consider all the factors relevant to frost formation and is regarded as an office companion for making frost forecasts.

Development of EDMI Calibration Baseline, HR-241, 1984

Electronic distance measuring instruments (EDMI) are used by surveyors in routine length measurements. The constant and scale factors of the instrument tend to change due to usage, transportation, and aging of crystals. Calibration baselines are established to enable surveyors to check the instruments and determine any changes in the values of constant and scale factors. The National Geodetic Survey (NGS) has developed guidelines for establishing these baselines. In 1981 an EDMI baseline at ISU was established according to NGS guidelines. In October 1982, the NGS measured the distances between monuments. Computer programs for reducing observed distances were developed. Mathematical model and computer programs for determining constant and scale factors were developed. A method was developed to detect any movements of the monuments. Periodic measurements of the baseline were made. No significant movement of the monuments was detected.

Iowa Development of Rubblized Concrete Pavement Base - Mills County, HR-315, 1995

Iowa counties have tried to rehabilitate deteriorating portland cement concrete (PCC) pavements with standard overlays, placement of engineering fabric, rock, open graded bituminous mixes and cracking and seating. While these methods prolong the life of the road, the cracks in the old pavement have eventually reflected to the surface. One possible alternative for rehabilitating severely deteriorated roads and preventing reflective cracking is the rubblization process. The objective of this research project was to rehabilitate and evaluate a severely deteriorated PCC roadway using a rubblization process. A 3.0 km (1.9 mi) section of L63 in Mills County was selected for this research. The road was divided into 16 sections. A resonate frequency vibration pavement breaker was used to rubblize the existing pavement. The variables of rubblization, drainage, and ACC overlay depths of 75 mm (3 in.), 100 mm (4 in.), and 125 mm (5 in.) were evaluated. The research on rubblized concrete pavement bases support the following conclusions: (1) The rubblization process prevents reflective cracking; (2) Edge drains improved the structural rating of the rubblized roadway; (3) An ACC overlay of 125 mm (5 in.) on a rubblized base provided an excellent roadway regardless of soil and drainage conditions; (4) An ACC overlay of 75 mm (3 in.) on a rubblized base can provide a good roadway if the soil structure below the rubblized base is stable and well drained; and (5) The Road Rater structural ratings of the rubblized test sections for this project are comparable to the nonrubblized test sections.

Development of Bio-Based Polymers for Use in Asphalt, TR-639, 2014

Asphalt binder is typically modified with poly type (styrene-butadiene-styrene or SBS) polymers to improve its rheological properties and performance grade. The elastic and principal component of SBS polymers is butadiene. For the last decade, butadiene prices have fluctuated and significantly increased, leading state highway agencies to search for economically viable alternatives to butadiene based materials. This project reports the recent advances in polymerization techniques that have enabled the synthesis of elastomeric, thermoplastic, block-copolymers (BCPs) comprised of styrene and soybean oil, where the “B” block in SBS polymers is replaced with polymerized triglycerides derived from soybean oil. These new breeds of biopolymers have elastomeric properties comparable to well-established butadiene-based styrenic BCPs. In this report, two types of biopolymer formulations are evaluated for their ability to modify asphalt binder. Laboratory blends of asphalt modified with the biopolymers are tested for their rheological properties and performance grade. Blends of asphalt modified with the biopolymers are compared to blends of asphalt modified with two commonly used commercial polymers. The viscoelastic properties of the blends show that biopolymers improve the performance grade of the asphalt to a similar and even greater extent as the commercial SBS polymers. Results shown in this report indicate there is an excellent potential for the future of these biopolymers as economically and environmentally favorable alternatives to their petrochemically-derived analogs.

Development of Self-Cleaning Box Culvert Design – Phase II, TR-619, 2013

Culverts are common means to convey flow through the roadway system for small streams. In general, larger flows and road embankment heights entail the use of multibarrel culverts (a.k.a. multi-box) culverts. Box culverts are generally designed to handle events with a 50-year return period, and therefore convey considerably lower flows much of the time. While there are no issues with conveying high flows, many multi-box culverts in Iowa pose a significant problem related to sedimentation. The highly erosive Iowa soils can easily lead to the situation that some of the barrels can silt-in early after their construction, becoming partially filled with sediment in few years. Silting can reduce considerably the capacity of the culvert to handle larger flow events. Phase I of this Iowa Highway Research Board project (TR-545) led to an innovative solution for preventing sedimentation. The solution was comprehensively investigated through laboratory experiments and numerical modeling aimed at screening design alternatives and testing their hydraulic and sediment conveyance performance. Following this study phase, the Technical Advisory Committee suggested to implement the recommended sediment mitigation design to a field site. The site selected for implementation was a 3-box culvert crossing Willow Creek on IA Hwy 1W in Iowa City. The culvert was constructed in 1981 and the first cleanup was needed in 2000. Phase II of the TR 545 entailed the monitoring of the site with and without the selfcleaning sedimentation structure in place (similarly with the study conducted in laboratory). The first monitoring stage (Sept 2010 to December 2012) was aimed at providing a baseline for the operation of the as-designed culvert. In order to support Phase II research, a cleanup of the IA Hwy 1W culvert was conducted in September 2011. Subsequently, a monitoring program was initiated to document the sedimentation produced by individual and multiple storms propagating through the culvert. The first two years of monitoring showed inception of the sedimentation in the first spring following the cleanup. Sedimentation continued to increase throughout the monitoring program following the depositional patterns observed in the laboratory tests and those documented in the pre-cleaning surveys. The second part of Phase II of the study was aimed at monitoring the constructed self-cleaning structure. Since its construction in December 2012, the culvert site was continuously monitored through systematic observations. The evidence garnered in this phase of the study demonstrates the good performance of the self-cleaning structure in mitigating the sediment deposition at culverts. Besides their beneficial role in sediment mitigation, the designed self-cleaning structures maintain a clean and clear area upstream the culvert, keep a healthy flow through the central barrel offering hydraulic and aquatic habitat similar with that in the undisturbed stream reaches upstream and downstream the culvert. It can be concluded that the proposed self-cleaning structural solution “streamlines” the area upstream the culvert in a way that secures the safety of the culvert structure at high flows while producing much less disturbance in the stream behavior compared with the current constructive approaches.

Development of a Conductometric Test for Frost Resistance of Concrete, HR-272, 1988

Research is described that was aimed at developing a test method which can be reasonably and rapidly performed in the laboratory and in the field to predict, with a high degree of certainty, the behavior of concrete subjected to the action of alternate freezing and thawing. The conductometric evaluation of concrete durability was explored with 3 different test methods: conductometric evaluation of the resistance of concrete to rapid freezing and thawing; conductomtric evaluation of the resistance of concrete to natural freezing and thawing, and conductometric evaluation of the pore size distribution of concrete and its correlation to concrete durability. The study showed that conductance could be used as a viable method for determining the durability of portland cement concrete. This would also allow the continuous monitoring of concrete durability without the removal twice per week from the freeze/thaw chamber. Recommendations for the continued development of these test methods are also included.

CTLA-4 controls the thymic development of both conventional and regulatory T cells through modulation of the TCR repertoire.

Cytotoxic T lymphocyte-associated antigen-4 (CTLA-4; CD152) is of pivotal importance for self-tolerance, with deficiency or unfavorable polymorphisms leading to autoimmune disease. Tolerance to self-antigens is achieved through thymic deletion of highly autoreactive conventional T (Tconv) cells and generation of FoxP3(+) regulatory T (Treg) cells. The main costimulatory molecule, CD28, augments the negative selection of Tconv cells and promotes the generation of FoxP3(+) Treg cells. The role of its antagonistic homolog CTLA-4, however, remains a topic of debate. To address this topic, we investigated the thymic development of T cells in the presence and absence of CTLA-4 in a T-cell receptor (TCR) transgenic mouse model specific for the myelin basic protein peptide Ac1-9. We reveal that CTLA-4 is expressed in the corticomedullary region of the thymus. Its absence alters the response of CD4(+)CD8(-) thymocytes to self-antigen recognition, which affects the quantity of the Treg cells generated and broadens the repertoire of peripheral Tconv cells. T-cell repertoire alteration after deletion of CTLA-4 results from changes in TCR Vα and Jα segment selection as well as CDR3α composition in Tconv and Treg cells. CTLA-4, therefore, regulates the early development of self-reactive T cells in the thymus and plays a key role in central tolerance.

Effects of maintaining ditch networks on the development of Scots pine stands

Tiivistelmä: Kunnostusojituksen vaikutus rämemänniköiden kehitykseen

Development of blast resistant somaclones of the upland rice cultivar araguaia

The degree of blast resistance of upland rice (Oryza sativa L.) cultivar Araguaia has decreased over time causing significant yield losses. The major objective of this study was to obtain blast (Pyricularia grisea) resistant somaclones, adapting greenhouse and field selection procedures. Rice blast resistance and agronomic traits were assessed in R2 to R6 generations derived from regenerant plants (R1) from immature panicles of Araguaia. The evaluation and selection procedures include testing of early segregating populations and fixed lines in the advanced generations, under natural field conditions, and artificial inoculations in the greenhouse, with prevalent races IB-1 and IB-9 of P. grisea. Somaclones with both vertical resistance and slow blasting resistance were obtained. Twenty of 31 somaclones developed with a high degree of vertical resistance and fan shaped plant type maintained resistance in field and blast nursery tests in the R6 generation. Greenhouse selection with two specific physiologic races yielded 44 somaclones with slow blasting resistance, similar plant type and yield potential as that of Araguaia.

Observational methodology in football: Development of an instrument to study the offensiva game in football.

The following paper introduces a new approach to the analysis of offensive game in football. Therefore, the main aim of this study was to create an instrument for collecting information for the analysis of offensive action and interactions game. The observation instrument that was used to accomplish the main objective of this work consists of a combination of format fields (FC) and systems of categories (SC). This methodology is a particular strategy of the scientific method that has as an objective to analyse the perceptible behaviour that occurs in habitual contexts, allowing them to be formally recorded and quantified and using an ad hoc instrument in order to obtain a behaviour systematic registration that, since they have been transformed in quantitative data with the necessary reliability and validity determined level, will allow analysis of the relations between these behaviours. The codifications undertaken to date in various games of football have shown that it serves the purposes for which it was developed, allowing more research into the offensive game methods in football.