Dimitrios G. Georgakopoulos

Resurgence in Ice Nuclei Measurement Research

Understanding cloud and precipitation responses to variations in atmospheric aerosols remains an important research topic for improving the prediction of climate. Knowledge is most uncertain, and the potential impact on climate is largest with regard to how aerosols impact ice formation in clouds. In this paper, we show that research on atmospheric ice nucleation, including the development of new measurement systems, is occurring at a renewed and historically unparalleled level. A historical perspective is provided on the methods and challenges of measuring ice nuclei, and the various factors that led to a lull in research efforts during a nearly 20-yr period centered about 30 yr ago. Workshops played a major role in defining critical needs for improving measurements at that time and helped to guide renewed efforts. Workshops were recently revived for evaluating present research progress. We argue that encouraging progress has been made in the consistency of measurements using the present generation of ic...

Biological control of cucumber and sugar beet damping-off caused by Pythium ultimum with bacterial and fungal antagonists.

Aims: Five bacterial strains belonging to Bacillus subtilis, Pseudomonas fluorescens and Ps. corrugata and two fungal strains belonging to Trichoderma viride and Gliocladium virens were evaluated for their efficacy in controlling sugar beet and cucumber damping‐off caused by Pythium ultimum.

Measurement of Ice Nucleation-Active Bacteria on Plants and in Precipitation by Quantitative PCR

ABSTRACT Ice nucleation-active (INA) bacteria may function as high-temperature ice-nucleating particles (INP) in clouds, but their effective contribution to atmospheric processes, i.e., their potential to trigger glaciation and precipitation, remains uncertain. We know little about their abundance on natural vegetation, factors that trigger their release, or persistence of their ice nucleation activity once airborne. To facilitate these investigations, we developed two quantitative PCR (qPCR) tests of the ina gene to directly count INA bacteria in environmental samples. Each of two primer pairs amplified most alleles of the ina gene and, taken together, they should amplify all known alleles. To aid primer design, we collected many new INA isolates. Alignment of their partial ina sequences revealed new and deeply branching clades, including sequences from Pseudomonas syringae pv. atropurpurea, Ps. viridiflava, Pantoea agglomerans, Xanthomonas campestris, and possibly Ps. putida, Ps. auricularis, and Ps. poae. qPCR of leaf washings recorded ∼108 ina genes g−1 fresh weight of foliage on cereals and 105 to 107 g−1 on broadleaf crops. Much lower populations were found on most naturally occurring vegetation. In fresh snow, ina genes from various INA bacteria were detected in about half the samples but at abundances that could have accounted for only a minor proportion of INP at −10°C (assuming one ina gene per INA bacterium). Despite this, an apparent biological source contributed an average of ∼85% of INP active at −10°C in snow samples. In contrast, a thunderstorm hail sample contained 0.3 INA bacteria per INP active at −10°C, suggesting a significant contribution to this sample.

Stabilization of human urine doping control samples.

Sport urine samples collected worldwide are not always shipped refrigerated from the collection sites to the World Anti-Doping Agency (WADA) Accredited Laboratories, especially when multiple days of transport are required [1,2]. Commensal urethral flora, urinary pathogens, and species of the environment may contaminate urine specimens [1,3–8] because sample collection procedures are not carried out under sterile conditions. As a result, enzymatic activities produced by microbial contamination may cause changes to the natural androgenic steroid profiles [1,4–10], nicking of human chorionic gonadotropin (hCG) molecule, and subsequent dissociation of gonadotropins [11–15] as well as shifting of erythropoietin (EPO) electrophoretic bands [16–18]. Also, during the past couple of years, antidoping science has been concerned about proteolytic enzymes, or alias proteases, added on purpose to distort the analysis results by degrading EPO [19–22]. The occurrence of degraded urine samples varies over the course of the year, with a peak during summer depending on the country where the specimens are collected as well as the duration of transportation [6,23,24]. If delays are unavoidable and refrigeration of samples is not possible, an effective method of preserving urine specimen is desirable. Up to the current time, no stabilization method has been applied in the urine collection material [2,3,6,20]. The purpose of this

Internet of Things Platform for Smart Farming: Experiences and Lessons Learnt

Improving farm productivity is essential for increasing farm profitability and meeting the rapidly growing demand for food that is fuelled by rapid population growth across the world. Farm productivity can be increased by understanding and forecasting crop performance in a variety of environmental conditions. Crop recommendation is currently based on data collected in field-based agricultural studies that capture crop performance under a variety of conditions (e.g., soil quality and environmental conditions). However, crop performance data collection is currently slow, as such crop studies are often undertaken in remote and distributed locations, and such data are typically collected manually. Furthermore, the quality of manually collected crop performance data is very low, because it does not take into account earlier conditions that have not been observed by the human operators but is essential to filter out collected data that will lead to invalid conclusions (e.g., solar radiation readings in the afternoon after even a short rain or overcast in the morning are invalid, and should not be used in assessing crop performance). Emerging Internet of Things (IoT) technologies, such as IoT devices (e.g., wireless sensor networks, network-connected weather stations, cameras, and smart phones) can be used to collate vast amount of environmental and crop performance data, ranging from time series data from sensors, to spatial data from cameras, to human observations collected and recorded via mobile smart phone applications. Such data can then be analysed to filter out invalid data and compute personalised crop recommendations for any specific farm. In this paper, we present the design of SmartFarmNet, an IoT-based platform that can automate the collection of environmental, soil, fertilisation, and irrigation data; automatically correlate such data and filter-out invalid data from the perspective of assessing crop performance; and compute crop forecasts and personalised crop recommendations for any particular farm. SmartFarmNet can integrate virtually any IoT device, including commercially available sensors, cameras, weather stations, etc., and store their data in the cloud for performance analysis and recommendations. An evaluation of the SmartFarmNet platform and our experiences and lessons learnt in developing this system concludes the paper. SmartFarmNet is the first and currently largest system in the world (in terms of the number of sensors attached, crops assessed, and users it supports) that provides crop performance analysis and recommendations.

Stabilization of human urine doping control samples: II. microbial degradation of steroids.

The transportation of urine samples, collected for doping control analysis, does not always meet ideal conditions of storage and prompt delivery to the World Anti-Doping Agency (WADA) accredited laboratories. Because sample collection is not conducted under sterile conditions, microbial activity may cause changes to the endogenous steroid profiles of samples. In the current work, funded by WADA, a chemical mixture consisting of antibiotics, antimycotic substances and protease inhibitors was applied in urine aliquots fortified with conjugated and deuterated steroids and inoculated with nine representative microorganisms. Aliquots with and without the chemical mixture were incubated at 37 degrees C for 7 days to simulate the transportation period, whereas another series of aliquots was stored at -20 degrees C as reference. Microbial growth was assessed immediately after inoculation and at the end of the incubation period. Variations in pH and specific gravity values were recorded. Gas chromatography-mass spectrometry (GC-MS) analysis was performed for the detection of steroids in the free, glucuronide, and sulfate fractions. The addition of the chemical stabilization mixture to urine samples inhibited microorganism growth and prevented steroid degradation at 37 degrees C. On the other hand, four of the nine microorganisms induced alterations in the steroid profile of the unstabilized samples incubated at 37 degrees C.

Stabilization of human urine doping control samples: a current opinion

Transportation of doping control urine samples from the collection sites to the World Anti-doping Agency (WADA) Accredited Laboratories is conducted under ambient temperatures. When sample delivery is not immediate, microbial contamination of urine, especially in summer, is a common phenomenon that may affect sample integrity and may result in misinterpretation of analytical data. Furthermore, the possibility of intentional contamination of sports samples during collection with proteolytic enzymes, masking the abuse of prohibited proteins such as erythropoietin (EPO) and peptide hormones, is a practice that has already been reported. Consequently, stabilization of urine samples with a suitable method in a way that protects samples’ integrity is important. Currently, no stabilization method is applied in the sample collection equipment system in order to prevent degradation of urine compounds. The present work is an overview of a study, funded by WADA, on degradation and stabilization aspects of sports urine samples against the above threats of degradation. Extensive method development resulted in the creation of a mixture of chemical agents for the stabilization of urine. Evaluation of results demonstrated that the stabilization mixture could stabilize endogenous steroids, recombinant EPO, and human chorionic gonadotropin in almost the entire range of the experimental conditions tested.

Development of a Fully Automated Flow Injection Analyzer Implementing Bioluminescent Biosensors for Water Toxicity Assessment

This paper describes the development of an automated Flow Injection analyzer for water toxicity assessment. The analyzer is validated by assessing the toxicity of heavy metal (Pb2+, Hg2+ and Cu2+) solutions. One hundred μL of a Vibrio fischeri suspension are injected in a carrier solution containing different heavy metal concentrations. Biosensor cells are mixed with the toxic carrier solution in the mixing coil on the way to the detector. Response registered is % inhibition of biosensor bioluminescence due to heavy metal toxicity in comparison to that resulting by injecting the Vibrio fischeri suspension in deionised water. Carrier solutions of mercury showed higher toxicity than the other heavy metals, whereas all metals show concentration related levels of toxicity. The biosensor’s response to carrier solutions of different pHs was tested. Vibrio fischeri’s bioluminescence is promoted in the pH 5–10 range. Experiments indicate that the whole cell biosensor, as applied in the automated fluidic system, responds to various toxic solutions.

Stabilization of human urine doping control samples: III. Recombinant human erythropoietin

BACKGROUND The tampering of athletes urine samples by the addition of proteolytic enzymes during the doping control sampling procedure was reported recently. The aim of the current study, funded by the World Anti-Doping Agency (WADA), was the application of a stabilization mixture in urine samples to chemically inactivate proteolytic enzymes and improve the electrophoteric signal of erythropoietin (EPO) in human urine. METHODS The stabilization mixture applied was a combination of antibiotics, antimycotic substances and protease inhibitors. A series of incubation experiments were conducted under controlled conditions in the presence and absence of the stabilization mixture in urine aliquots spiked with six proteases. Two different analytical techniques were applied for the qualitative and quantitative EPO measurement: isoelectric focusing (IEF) and chemiluminescent immunoassay respectively. RESULTS The addition of the chemical stabilization mixture into urine aliquots substantially improved EPO detection in the presence of proteolytic enzymes following incubation at 37 degrees C or storage at -20 degrees C. CONCLUSIONS The results of this study indicated that the stabilization of urine prior to the sample collection procedure with the proposed chemical mixture might prove to be a useful tool for the preservation of anti-doping samples.

Functional characterization of NopT1 and NopT2, two type III effectors of Bradyrhizobium japonicum

NopT1 and NopT2, putative type III effectors from the plant symbiotic bacterium Bradyrhizobium japonicum, are predicted to belong to a family of YopT/AvrPphB effectors, which are cysteine proteases. In the present study, we showed that both NopT1 and NopT2 indeed possess cysteine protease activity. When overexpressed in Escherichia coli, both NopT1 and NopT2 undergo autoproteolytic processing which is largely abolished in the presence of E-64, a papain family-specific inhibitor. Mutations of NopT1 disrupting either the catalytic triad or the putative autoproteolytic site reduce or markedly abolish the protease activity. Autocleavage likely occurs between residues K48 and M49, though another potential cleavage site is also possible. NopT1 also elicitis HR-like cell death when transiently expressed in tobacco plants and its cysteine protease activity is essential for this ability. In contrast, no macroscopic symptoms were observed for NopT2. Furthermore, mutational analysis provided evidence that NopT1 may undergo acylation inside plant cells and that this would be required for its capacity to elicit HR-like cell death in tobacco.