Bacterial sensors: synthetic design and application principles

Bacterial reporters are live, genetically engineered cells with promising application in bioanalytics. They contain genetic circuitry to produce a cellular sensing element, which detects the target compound and relays the detection to specific synthesis of so-called reporter proteins (the presence or activity of which is easy to quantify). Bioassays with bacterial reporters are a useful complement to chemical analytics because they measure biological responses rather than total chemical concentrations. Simple bacterial reporter assays may also replace more costly chemical methods as a first line sample analysis technique. Recent promising developments integrate bacterial reporter cells with microsystems to produce bacterial biosensors. This lecture presents an in-depth treatment of the synthetic biological design principles of bacterial reporters, the engineering of which started as simple recombinant DNA puzzles, but has now become a more rational approach of choosing and combining sensing, controlling and reporting DNA 'parts'. Several examples of existing bacterial reporter designs and their genetic circuitry will be illustrated. Besides the design principles, the lecture also focuses on the application principles of bacterial reporter assays. A variety of assay formats will be illustrated, and principles of quantification will be dealt with. In addition to this discussion, substantial reference material is supplied in various Annexes.

Back Warning Sensors for Tow Plows, RB07-012, 2013

Backup warning system devices were evaluated to determine if they would alert winter maintenance snow plow drivers to obstacles directly behind the trailer and out of view of the driver when a unit is backed up. When the sensors on the back of the tow plow were covered with snow during plowing operations, the sensor would go off in the cab and continue going off, which would result in drivers turning the volume of the unit way down. One shop stated that the wireless transmitted signal would be hit or miss depending on the winter weather that they were operating in. The sensors on the back of the tow plow trailer would come in contact with salt brine and in this situation one of the sensors did go bad. The weatherproof box that was designed to keep the system waterproof did not fully keep the moisture out. It was found that the system did alert drivers of items behind the unit and there were no backup accidents reported during the research period.

Optimization Of Promoter Sequences In Order To Mimic Physiological Pde6b Expression

Purpose: Many retinal degenerations result from defective retina-specific gene expressions. Thus, it is important to understand how the expression of a photoreceptor-specific gene is regulated in vivo in order to achieve successful gene therapy. The present study aims to design an AAV2/8 vector that can regulate the transcript level in a physiological manner to replace missing PDE6b in Rd1 and Rd10 mice. In previous studies (Ogieta, et al., 2000), the short 5' flanking sequence of the human PDE6b gene (350 bp) was shown to be photoreceptor-specific in transgenic mice. However, the efficiency and specificity of the 5' flanking region of the human PDE6b was not investigated in the context of gene therapy during retinal degeneration. In this study, two different sequences of the 5' flanking region of the human PDE6b gene were studied as promoter elements and their expression will be tested in wild type and diseased retinas (Rd 10 mice).Methods: Two 5' flanking fragments of the human PDE6b gene: (-93 to +53 (150 bp) and -297 to +53 (350 bp)) were cloned in different plasmids in order to check their expression in vitro and in vivo by constructing an AAV2/8 vector. These elements drove the activity of either luciferase (pGL3 plasmids) or EGFP. jetPEI transfection in Y 79 cells was used to evaluate gene expression through luciferase activity. Constructs encoding EGFP under the control of the two promoters were performed in AAV2.1-93 (or 297)-EGFP plasmids to produce AAV2/8 vectors.Results: When pGL3-93 (150 bp) or pGL3-297 (350 bp) were transfected in the Y-79 cells, the smaller fragment (150 bp) showed higher gene expression compared to the 350 bp element and to the SV40 control, as previously reported. The 350 bp drove similar levels of expression when compared to the SV40 promoter. In view of these results, the fragments (150 bp or 350 bp) were integrated into the AAV2.1-EGFP plasmid to produce AAV2/8 vector, and we are currently evaluating the efficiency and specificity of the produced constructs in vivo in normal and diseased retinas.Conclusions: Comparisons of these vectors with vectors bearing ubiquitous promoters should reveal which construct is the most suitable to drive efficient and specific gene expression in diseased retinas in order to restore a normal function on the long term.

On the complementariness of infrasound and seismic sensors for monitoring snow avalanches

The paper analyses and compares infrasonic and seismic data from snow avalanches monitored at the Vallée de la Sionne test site in Switzerland from 2009 to 2010. Using a combination of seismic and infrasound sensors, it is possible not only to detect a snow avalanche but also to distinguish between the different flow regimes and to analyse duration, average speed (for sections of the avalanche path) and avalanche size. Different sensitiveness of the seismic and infrasound sensors to the avalanche regimes is shown. Furthermore, the high amplitudes observed in the infrasound signal for one avalanche were modelled assuming that the suspension layer of the avalanche acts as a moving turbulent sound source. Our results show reproducibility for similar avalanches on the same avalanche path.

Physiological responses of two tropical weeds to shade: I. Growth and biomass allocation

Ipomoea asarifolia (Desr.) Roem. & Schultz (Convolvulaceae) and Stachytarpheta cayennensis (Rich) Vahl. (Verbenaceae), two weeds found in pastures and crop areas in Brazilian Amazonia, were grown in controlled environment cabinets under high (800-1000 µmol m-² s-¹) and low (200-350 µmol m-² s-¹) light regimes during a 40-day period. For both species leaf dry mass and leaf area per total plant dry mass, and leaf area per leaf dry mass were higher for low-light plants, whereas root mass per total plant dry mass was higher for high-light plants. High-light S. cayennensis allocated significantly more biomass to reproductive tissue than low-light plants, suggesting a probably lower ability of this species to maintain itself under shaded conditions. Relative growth rate (RGR) in I. asarifolia was initially higher for high-light grown plants and after 20 days started decreasing, becoming similar to low-light plants at the last two harvests (at 30 and 40 days). In S. cayennensis, RGR was also higher for high-light plants; however, this trend was not significant at the first and last harvest dates (10 and 40 days). These results are discussed in relation to their ecological and weed management implications.

Physiological responses of two tropical weeds to shade: II. Leaf gas exchange and nitrogen content

Ipomoea asarifolia (Desr.) Roem. & Schultz (Convolvulaceae) and Stachytarpheta cayennensis (Rich) Vahl. (Verbenaceae), two weeds found in pastures and crop areas in the Brazilian Amazonia, Brazil, were grown in controlled environment cabinets under high (800-1000 µmol m-² s-¹) and low (200-350 µmol m-² s-¹) light regimes during a 40-day period. The objective was to determine the effect of shade on photosynthetic features and leaf nitrogen content of I. asarifolia and S. cayennensis. High-irradiance grown I. asarifolia leaves had significantly higher dark respiration and light saturated rates of photosynthesis than low-irradiance leaves. No significant differences for these traits, between treatments, were observed in S. cayennensis. Low-irradiance leaves of both species displayed higher CO2 assimilation rates under low irradiance. High-irradiance grown leaves of both species had less nitrogen per unit of weight. Low-irradiance S. cayennensis had more nitrogen per unit of leaf area than high-irradiance plants; however, I. asarifolia showed no consistent pattern for this variable through time. For S. cayennensis, leaf nitrogen content and CO2 assimilation were inversely correlated to the amount of biomass allocated to developing reproductive structures. These results are discussed in relation to their ecological and weed management implications.

Physiological changes during postharvest senescence of broccoli

The objective of this work was to determine the early physiological changes throughout shelf life of fresh broccoli (Brassica oleracea L. var. italica) cv. Piracicaba Precoce at 25ºC and relative humidity of 96% in the dark until complete senescence. Head inflorescences showed lack of turgidity and commercial value when weight loss reached up to 5%, coinciding with 48 hour after harvest. Chlorophyll content was stable until 24 hours after harvesting; afterwards, an intense degradation phase took place. At 72 hours, total head yellowing was observed when chlorophyll content dropped to 30% of its initial content. Peroxidase activity increased by 1.4 fold during the first six hours, dropping to its lowest level approximately 24 hours after harvesting. However, from this time on, an increment of activity was observed until 72 hours. At 24 hours after harvesting, respiration was reduced by 50%. At later stages of senescence, respiration of florets was stable, but in a lower level than that determined at harvest. Sharp reduction of starch and reducing sugars was observed within 24 hours after harvesting, followed by continuous period of decline in starch and non-reducing sugars.

Physiological responses of maize and cowpea to intercropping

The effect of intercropping on plant water status, gas exchange and productivity of maize (Zea mays L.) cv. Centralmex, and cowpea (Vigna unguiculata L. (Walp)) cv. Pitiuba were evaluated under semi-arid conditions at the Embrapa-Centro de Pesquisa Agropecuária do Trópico Semi-Árido (CPATSA) at Petrolina, PE, Brazil. The treatments were: maize and cowpea as sole crops, at a population of 40,000 plants ha-1, and intercropped at a population of 20,000 plants ha-1. The results obtained in this paper appear to be related to the degree of competition experienced by the components, mainly for water and light. Maize intercropped had higher values of leaf water potential, stomatal conductance, transpiration and photosynthesis than as sole crop. Intercropped cowpea had higher values of leaf water potential but lower stomatal conductance, transpiration and photosynthesis than sole cowpea. Maize productivity increased 18% in relation to sole crop whereas a 5% decrease was observed with cowpea. Despite these facts the Land Equivalent Ratio obtained was 1.13 indicating intercropping advantage over the sole system. The higher partial Land Equivalent Ratio observed for maize suggests that this specie was the main component influencing the final productivity of the intercropping system studied.

Physiological and morphological responces of Brachiaria spp. to flooding

The physiological and morphological responses of the forage grasses Brachiaria brizantha cv. Marandu, B. decumbens and B. humidicola were compared for plants grown in pots under flooding and well-drained conditions for 14 days. Flooding reduced specific leaf area and biomass allocation to roots in all species and enhanced leaf senescence in B. brizantha and B. decumbens. Relative growth rate was reduced by flooding in B. brizantha and B. decumbens, but not in B. humidicola.Leaf elongation rate was unaffected by flooding in B. decumbens and B. humidicola, but declined in B. brizantha since the first day of flooding. Net photosynthesis and leaf chlorophyll content were reduced by flooding in B. brizantha; however, no flooding effect could be detected in the other two species. For all species, there was a close relationship between net photosynthesis and stomatal conductance under flooding. These results show that the studied species have distinct degrees of tolerance to flood, B. brizantha is intolerant, B. decumbens is moderately tolerant and B. humidicola is tolerant. Because leaf elongation rate was immediately depressed by flooding only in B. brizantha, this measurement could be appropriate as an early detection mechanism for relative flood tolerance in Brachiaria spp.

A Feasibility Study on Embedded Micro-Electromechanical Sensors and Systems (MEMS) for Monitoring Highway Structures, TR-575, 2011

Micro-electromechanical systems (MEMS) provide vast improvements over existing sensing methods in the context of structural health monitoring (SHM) of highway infrastructure systems, including improved system reliability, improved longevity and enhanced system performance, improved safety against natural hazards and vibrations, and a reduction in life cycle cost in both operating and maintaining the infrastructure. Advancements in MEMS technology and wireless sensor networks provide opportunities for long-term continuous, real-time structural health monitoring of pavements and bridges at low cost within the context of sustainable infrastructure systems. The primary objective of this research was to investigate the use of MEMS in highway structures for health monitoring purposes. This study focused on investigating the use of MEMS and their potential applications in concrete through a comprehensive literature review, a vendor survey, and a laboratory study, as well as a small-scale field study. Based on the comprehensive literature review and vendor survey, the latest information available on off-the-shelf MEMS devices, as well as research prototypes, for bridge, pavement, and traffic applications were synthesized. A commercially-available wireless concrete monitoring system based on radio-frequency identification (RFID) technology and off-the-shelf temperature and humidity sensors were tested under controlled laboratory and field conditions. The test results validated the ability of the RFID wireless concrete monitoring system in accurately measuring the temperature both inside the laboratory and in the field under severe weather conditions. In consultation with the project technical advisory committee (TAC), the most relevant MEMS-based transportation infrastructure research applications to explore in the future were also highlighted and summarized.

Effects of group size and early handling on some behavioural and physiological welfare parameters in farmed blue foxes

Selostus: Ryhmäkoon ja varhaisen käsittelyn vaikutus tarhattujen sinikettujen hyvinvointiin

Effects of group size and space allocation on physiological, behavioural and production-related welfare parameters in farmed silver fox cubs

Selostus: Ryhmäkoon ja käytössä olevan tilan vaikutus tarhattujen hopeakettupentujen hyvinvointiin

Affective and physiological responses to environmental noises and music

Research suggests that respiratory patterns may reflect general dimensions of emotional response. In this study, we investigated the relationships between judgments of affective valence (pleasantness) and arousal and respiratory responses to acoustic stimuli. Sixteen environmental noises and 16 musical fragments of 30 s duration were presented to 31 participants, while respiration, skin conductance level and heart rate were recorded. Judgments of valence and arousal were registered using the 9-point Self-Assessment Manikin. For noises, breathing accelerated and minute ventilation augmented with decreases in pleasantness for low-arousal stimuli and with increases in arousal for positive stimuli. For music, breathing accelerated and minute ventilation augmented with increases both in rated valence and arousal. Skin conductance level increased with arousal ratings for music but not for noises, whereas mean heart rate increased with rated arousal for noises but not for music. Although both noises and music are sound-vibrations, differences in the relationships between affective judgments and physiological responses were found suggesting differences in the processing of the two types of acoustic stimuli. [Authors]