Monitoring of Peronospora destructor Primary and Secondary Inoculum by Real-Time qPCR

Onion downy mildew (ODM), caused by Peronospora destructor, is a serious threat for onion growers worldwide. In southwestern Québec, Canada, a steady increase in occurrence of ODM has been observed since the mid-2000s. On onion, P. destructor can develop local and systemic infections producing numerous sporangia which act as initial inoculum locally and also for neighboring areas. It also produces oospores capable of surviving in soils and tissues for a prolonged period of time. A recent study showed that ODM epidemics are strongly associated with weather conditions related to production and survival of overwintering inoculum, stressing the need to understand the role of primary (initial) and secondary inoculum. However, P. destructor is an obligate biotrophic pathogen, which complicates the study of inoculum sources. This study aimed at developing a molecular assay specific to P. destructor, allowing its quantification in environmental samples. In this study, a reliable and sensitive hydrolysis probe-based assay multiplexed with an internal control was developed on the internal transcribed spacer (ITS) region to quantify soil- and airborne inoculum of P. destructor. The assay specificity was tested against 17 isolates of P. destructor obtained from different locations worldwide, other members of the order Peronosporales, and various onion pathogens. Validation with artificially inoculated soil and air samples suggested a sensitivity of less than 10 sporangia g−1 of dry soil and 1 sporangium m−3 of air. Validation with environmental air samples shows a linear relationship between microscopic and real-time quantitative PCR counts. In naturally infested soils, inoculum ranged from 0 to 162 sporangia equivalent g−1 of dry soil, which supported the hypothesis of overwintering under northern climates. This assay will be useful for primary and secondary inoculum monitoring to help characterize ODM epidemiology and could be used for daily tactical and short-term strategic decision-making.

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Detection and Quantification of Pythium tracheiphilum in Soil by Multiplex Real-Time qPCR

Plant Disease ◽

10.1094/pdis-03-18-0419-re ◽

2019 ◽

Vol 103 (3) ◽

pp. 475-483 ◽

Cited By ~ 6

Author(s):

Hervé Van der Heyden ◽

Thérèse Wallon ◽

C. André Lévesque ◽

Odile Carisse

Keyword(s):

Real Time ◽

Internal Control ◽

Disease Incidence ◽

Quantitative Polymerase Chain Reaction ◽

Pcr Primers ◽

Qpcr Assay ◽

Inoculum Concentration ◽

Soil P ◽

Dry Soil ◽

Detection And Quantification

In Canada, head lettuce (Lactuca sativa capitata) is extensively produced in the muck soils of southwestern Québec. However, yields are increasingly affected by various soilborne pathogens, including Pythium spp., which cause wilt and damping off. In a survey conducted in Québec muck soils in 2010 and 2011, Pythium tracheiphilum Matta was identified as the predominant Pythium sp. in the root of head lettuce showing Pythium stunt symptoms. Therefore, to improve risk assessment and help further understanding of disease epidemiology, a specific and sensitive real-time quantitative polymerase chain reaction (qPCR) assay based on TaqMan-minor groove binder (MGB) technology was developed for P. tracheiphilum. The PCR primers along with a TaqMan-MGB probe were designed from the ribosomal internal transcribed spacer 2 region. A 100-bp product was amplified by PCR from all P. tracheiphilum isolates tested while no PCR product was obtained from 38 other Pythium spp. or from a selection of additional lettuce pathogens tested. In addition to P. tracheiphilum, the assay was multiplexed with an internal control allowing for the individual validation of each PCR. In artificially infested soils, the sensitivity of the qPCR assay was established as 10 oospores/g of dry soil. P. tracheiphilum was not detected in soils in which lettuce has never been grown; however, inoculum ranged from 0 to more than 200,000 oospores/g of dry soil in commercial lettuce fields. Also, disease incidence was positively correlated with inoculum concentration (r = 0.764). The results suggest that inoculum concentration should be considered when making Pythium stunt management decisions. The developed qPCR assay will facilitate reliable detection and quantification of P. tracheiphilum from field soil.

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REAL CONDITION AND PROSPECTS OF DEVELOPMENT OF THE RADAR STATIONS OF THE COUNTER BATTERY FIGHTINGV

Collection of scientific works of Odesa Military Academy ◽

10.37129/2313-7509.2019.12.1.30-40 ◽

2019 ◽

Vol 1 (12) ◽

pp. 30-40

Author(s):

V. Нolovan ◽

V. Gerasimov ◽

А. Нolovan ◽

N. Maslich

Keyword(s):

Comparative Analysis ◽

Real Time ◽

Weather Conditions ◽

Time Of Day ◽

Current Position ◽

Real Condition ◽

Leading Role ◽

Artillery Shell

Fighting in the Donbas, which has been going on for more than five years, shows that a skillful counter-battery fight is an important factor in achieving success in wars of this kind. Especially in conditions where for the known reasons the use of combat aviation is minimized. With the development of technical warfare, the task of servicing the counter-battery fight began to rely on radar stations (radar) to reconnaissance the positions of artillery, which in modern terms are called counter-battery radar. The principle of counter-battery radar is based on the detection of a target (artillery shell, mortar mine or rocket) in flight at an earlier stage and making several measurements of the coordinates of the current position of the ammunition. According to these data, the trajectory of the projectile's flight is calculated and, on the basis of its prolongation and extrapolation of measurements, the probable coordinates of the artillery, as well as the places of ammunition falling, are determined. In addition, the technical capabilities of radars of this class allow you to recognize the types and caliber of artillery systems, as well as to adjust the fire of your artillery. The main advantages of these radars are:  mobility (transportability);  inspection of large tracts of terrain over long distances;  the ability to obtain target's data in near real-time;  independence from time of day and weather conditions;  relatively high fighting efficiency. The purpose of the article is to determine the leading role and place of the counter-battery radar among other artillery instrumental reconnaissance tools, to compare the combat capabilities of modern counter-battery radars, armed with Ukrainian troops and some leading countries (USA, China, Russia), and are being developed and tested in Ukraine. The method of achieving this goal is a comparative analysis of the features of construction and combat capabilities of modern models of counter-battery radar in Ukraine and in other countries. As a result of the conducted analysis, the directions of further improvement of the radar armament, increasing the capabilities of existing and promising counter-battery radar samples were determined.

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Abstract 343: Chest Compression Compliance With Guidelines Declines Over the Course of Rescues

Circulation ◽

10.1161/circ.140.suppl_2.343 ◽

2019 ◽

Vol 140 (Suppl_2) ◽

Author(s):

James K Russell ◽

Digna González Otero ◽

Sofia Ruiz de Gauna ◽

Mikel Leturiondo ◽

Jesus M Ruiz ◽

...

Keyword(s):

Real Time ◽

Chest Compression ◽

Steady Increase ◽

Release Force

Introduction: CPR guidelines recommend limits for rate and depth, and release (avoidance of leaning). Simultaneous compliance with all three recommendations is challenging, sustained compliance more so. Hypothesis: Compliance with rate, depth and release guidelines is limited, and will decline over the course of rescues. Methods: Acceleration and force signals were extracted from monitor/defibrillators equipped with CPR monitors in 248 adult cases of manual CPR during out-of-hospital resuscitations treated by Tualatin Valley Fire & Rescue (TVF&R, Tigard, OR) during 2016 and 2017. TVF&R personnel delivered continuous compression CPR with real-time feedback available for rate, depth and leaning. Depth was calculated from acceleration. Peak depth, release force, and rate were measured for each compression. Release with force exceeding 2.5 kg-f was counted as incomplete (leaning). Results: Cases included 1802 (1055-2683) (median, IQR) compressions (total 481,407 compressions). Compliance with all three recommendations was 25% initially, declining to 19% for the ends of the longest rescues (> 3000 compressions from 48 cases). Depth compliance and complete release varied relatively little (31-36%, 93-95% respectively) and irregularly over the course of resuscitation. Early, 45% of compressions were too shallow (< 50 mm), and 21% were too deep (> 60 mm). Depth exceptions shifted over the course of resuscitation (to 36%, 28% respectively). Rate compliance declined from 73% to 54%. Rates below 100 cpm were 9% initially, declining to 5%. Rates exceeding 120 cpm were 18% early on, but climbed to 41% towards the end of long resuscitations, accounting for most of the decline in compliance with guidelines. Conclusions: Compliance with all 3 recommendations for CPR compressions is challenging, and becomes more so as the chest changes in response to compressions. Most problematic is a steady increase in compression rates above 120 cpm.

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How Can Existing Ground-Based Profiling Instruments Improve European Weather Forecasts?

Bulletin of the American Meteorological Society ◽

10.1175/bams-d-17-0231.1 ◽

2019 ◽

Vol 100 (4) ◽

pp. 605-619 ◽

Cited By ~ 14

Author(s):

A. J. Illingworth ◽

D. Cimini ◽

A. Haefele ◽

M. Haeffelin ◽

M. Hervo ◽

...

Keyword(s):

Real Time ◽

Traffic Control ◽

Positive Impact ◽

Weather Forecast ◽

Weather Conditions ◽

Time Data ◽

Weather Forecasts ◽

Forecast Models ◽

Wind Lidar ◽

Microwave Radiometers

Abstract To realize the promise of improved predictions of hazardous weather such as flash floods, wind storms, fog, and poor air quality from high-resolution mesoscale models, the forecast models must be initialized with an accurate representation of the current state of the atmosphere, but the lowest few kilometers are hardly accessible by satellite, especially in dynamically active conditions. We report on recent European developments in the exploitation of existing ground-based profiling instruments so that they are networked and able to send data in real time to forecast centers. The three classes of instruments are i) automatic lidars and ceilometers providing backscatter profiles of clouds, aerosols, dust, fog, and volcanic ash, the last two being especially important for air traffic control; ii) Doppler wind lidars deriving profiles of wind, turbulence, wind shear, wind gusts, and low-level jets; and iii) microwave radiometers estimating profiles of temperature and humidity in nearly all weather conditions. The project includes collaboration from 22 European countries and 15 European national weather services, which involves the implementation of common operating procedures, instrument calibrations, data formats, and retrieval algorithms. Currently, data from 265 ceilometers in 19 countries are being distributed in near–real time to national weather forecast centers; this should soon rise to many hundreds. One wind lidar is currently delivering real time data rising to 5 by the end of 2019, and the plan is to incorporate radiometers in 2020. Initial data assimilation tests indicate a positive impact of the new data.

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Comparison and Evaluation of Organizational Transactions for Continuous Auditing and Business Compliance

International Journal of Information System Modeling and Design ◽

10.4018/ijismd.2018040101 ◽

2018 ◽

Vol 9 (2) ◽

pp. 1-23

Author(s):

Rui Pedro Marques ◽

Henrique Santos ◽

Carlos Santos

Keyword(s):

Real Time ◽

Internal Control ◽

Control Mechanisms ◽

Continuous Auditing ◽

Technical Details ◽

Ontological Model ◽

High Level ◽

Execution Pattern

This article presents a comparator module which aims to compare, in real time, executions of organizational transactions with patterns of behaviors of these transaction executions, allowing the determination of which execution pattern is being followed by running each transaction. This is according to information received by the internal control mechanisms, which continuously monitors the transaction executions. A possible application using this module was deployed and results were obtained from a case study. The results prove effectiveness of the module, mainly because it is able to assess business compliance and the qualitative risk associated to each transaction execution while it is running, enabling an efficient continuous auditing application. The innovation of this article is ensured by the use of an ontological model to represent organizational transactions, which can be applicable to any type of transaction in any business area in order to audit transactions at a very low level, contrary to what happens in traditional auditing, which occurs at a high level (e.g. compare whether a completed transaction has followed a set of procedures). Besides the conceptualization, this work presents some technical details of development and discussion of results from the case study.

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IoTSAS: An Integrated System for Real-Time Semantic Annotation and Interpretation of IoT Sensor Stream Data

Computers ◽

10.3390/computers10100127 ◽

2021 ◽

Vol 10 (10) ◽

pp. 127

Author(s):

Besmir Sejdiu ◽

Florije Ismaili ◽

Lule Ahmedi

Keyword(s):

Air Quality ◽

Real Time ◽

Contextual Information ◽

Semantic Annotation ◽

Weather Conditions ◽

Water Quality Monitoring ◽

Integrated System ◽

Quality Monitoring ◽

Stream Data ◽

Semantic Annotations

Sensors and other Internet of Things (IoT) technologies are increasingly finding application in various fields, such as air quality monitoring, weather alerts monitoring, water quality monitoring, healthcare monitoring, etc. IoT sensors continuously generate large volumes of observed stream data; therefore, processing requires a special approach. Extracting the contextual information essential for situational knowledge from sensor stream data is very difficult, especially when processing and interpretation of these data are required in real time. This paper focuses on processing and interpreting sensor stream data in real time by integrating different semantic annotations. In this context, a system named IoT Semantic Annotations System (IoTSAS) is developed. Furthermore, the performance of the IoTSAS System is presented by testing air quality and weather alerts monitoring IoT domains by extending the Open Geospatial Consortium (OGC) standards and the Sensor Observations Service (SOS) standards, respectively. The developed system provides information in real time to citizens about the health implications from air pollution and weather conditions, e.g., blizzard, flurry, etc.

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TLR, RAGE, and HMGB1 in the Inflammatory Response in Ph(-) Myeloproliferative Neoplasm

Blood ◽

10.1182/blood-2020-134988 ◽

2020 ◽

Vol 136 (Supplement 1) ◽

pp. 31-32

Author(s):

Guanfang Shi ◽

Kiron Nair ◽

Preethi Ramachandran ◽

Chi Chen ◽

Ching Wong ◽

...

Keyword(s):

Inflammatory Response ◽

Real Time ◽

Inflammatory Cytokines ◽

Internal Control ◽

Real Time Pcr ◽

Mononuclear Cells ◽

Conflicts Of Interest ◽

Philadelphia Chromosome ◽

Myeloproliferative Neoplasm ◽

Significant Difference

Recent evidence of increased constitutional symptoms and inflammatory cytokines in Philadelphia chromosome negative (Ph (-)) MPN suggests that an inflammatory response is important in the pathogenesis of Ph (-) MPN. Toll-like receptors (TLR), Receptor for Advanced Glycation End products (RAGE) and High mobility group protein B1 (HMGB1) are the important pathways for the inflammatory response. All these three important pathway proteins were studied in MPN diseases in the current studies. Materials and Methods: TLR assay. TLR 2,3, 4, 7, 9 quantification was performed by immuno-staining of 1×106 mononuclear cells (peripheral blood) which were incubated with fluorescence-conjugated anti-TLR-2,3, 4, 7, 9 antibodies and assayed by flow cytometry. HMGB1assay:HMGB1 ELISA kit from Immuno-Biological Laboratories, Inc. (IBL-America) were used. The plasma samples were diluted four times with the provided sample dilution buffer, and assayed in duplicate according to the manufacturer's suggestion. RAGE (RT-PCR) Assay: Total RNA was extracted from normal control or patient mononuclear cells. Predesigned primers for RAGE, and internal control genes were ordered from Qiagen (Germantown, MD). Real-time PCR was performed using SsoAdvanced™ Universal SYBR® Green Supermix (Bio-Rad, Hercules, CA) on Bio-Rad iQ5 Multicolor Real-Time PCR Detection System. At least three house-keeping genes (ribosomal protein L4, TATA box binding protein, and tubulin-α 1b) were used as normalization controls. The expression of RAGE were compared with each internal control. Average of three was used to calculate the ratio of final patient to normal Results: Total of 97 patients with MPN were studied 1) TLR: TLR 3,7,9 was not significantly different from controls. But TLR 2 was significantly increased in both PV, as well as in the MPN group when PV, ET and MF were grouped together as MPN (Fig A). TLR 4 was not significantly increased in PV, ET, MF individually but was found to be significantly increased than the controls, when they are grouped together as MPN (Fig B). 2) RAGE: No significant difference was found between ET, PV, MF individually or when they were grouped together as MPN than the controls (Fig C). 3) HMGB1: No significant difference was seen between ET, PV, MF or when they were grouped as MPN (Fig D). Conclusion: Current study suggests that TLR pathway especially TLR2, and to a lesser extent TLR4 are the important pathways for inflammatory response with increased inflammatory cytokines in MPN, while HMGB1 and RAGE pathways were not different from controls. Figure Disclosures No relevant conflicts of interest to declare.

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An IoT based Low-cost Weather Monitoring System for Smart Farming

Agricultural Science Digest - A Research Journal ◽

10.18805/ag.d-370 ◽

2021 ◽

Author(s):

L.P.S.S.K. Dayananda ◽

A. Narmilan ◽

P. Pirapuraj

Keyword(s):

Liquid Crystal ◽

Mobile Phone ◽

Real Time ◽

Monitoring System ◽

Liquid Crystal Display ◽

Low Cost ◽

Weather Conditions ◽

Weather Data ◽

The Internet ◽

Arduino Uno

Background: Weather monitoring is an important aspect of crop cultivation for reducing economic loss while increasing productivity. Weather is the combination of current meteorological components, such as temperature, wind direction and speed, amount and kind of precipitation, sunshine hours and so on. The weather defines a time span ranging from a few hours to several days. The periodic or continuous surveillance or the analysis of the status of the atmosphere and the climate, including parameters such as temperature, moisture, wind velocity and barometric pressure, is known as weather monitoring. Because of the increased usage of the internet, weather monitoring has been upgraded to smart weather monitoring. The Internet of Things (IoT) is one of the new technology that can help with many precision farming operations. Smart weather monitoring is one of the precision agriculture technologies that use sensors to monitor correct weather. The main objective of the research is to design a smart weather monitoring and real-time alert system to overcome the issue of monitoring weather conditions in agricultural farms in order for farmers to make better decisions. Methods: Different sensors were used in this study to detect temperature and humidity, pressure, rain, light intensity, CO2 level, wind speed and direction in an agricultural farm and real time clock sensor was used to measured real time weather data. The major component of this system was an Arduino Uno microcontroller and the system ran according to a program written in the Arduino Uno software. Result: This is a low-cost smart weather monitoring system. This system’s output unit were a liquid crystal display and a GSM900A module. The weather data was displayed on a liquid crystal display and the GSM900A module was used to send the data to a mobile phone. This smart weather station was used to monitor real-time weather conditions while sending weather information to the farmer’s mobile phone, allowing him to make better decisions to increase yield.

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The Potential Benefits Of Real-Time Monitoring Of Combined Sewer Overflows In The City Of Toronto

10.32920/ryerson.14661741 ◽

2021 ◽

Author(s):

Romak Kordnejad

Keyword(s):

Real Time ◽

Weather Conditions ◽

Future Research ◽

Environmental Damage ◽

Real Time Monitoring ◽

Combined Sewer Overflows ◽

Combined Sewer ◽

Potential Benefits ◽

Sewer Overflows ◽

The City

This thesis focuses on the current state of combined sewer overflows (CSO) in the City of Toronto and the potential benefits of utilizing real-time monitoring system to explore options for optimizing current models and reducing future overflows. CSOs containing sanitary and stormwater sewage over spill during wet weather conditions, contaminating bodies of water. Antiquated sewer systems built to support population at the time, no longer have the required capacity to support exponential growth causing environmental damage. Key research findings include exploring the current CSO state in the City of Toronto, remediation plans executed by the City in hopes to reduce CSO and finally concluding the City’s progress to date. Extensive research on CSOs in North American cities has shown initiative towards reducing CSOs while using real-time monitoring systems which can be utilized in the City of Toronto. Future research is required to find effective and efficient ways of implementing such systems.

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Identification and evaluation of an appropriate housekeeping gene for real-time gene profiling of hepatocellular carcinoma cells cultured in three-dimensional scaffold

10.21203/rs.3.rs-490561/v1 ◽

2021 ◽

Author(s):

Anjana Kaveri Badekila ◽

Praveen Rai ◽

Sudarshan Kini

Keyword(s):

Hepatocellular Carcinoma ◽

Cell Culture ◽

Real Time ◽

Internal Control ◽

18S Rrna ◽

Three Dimensional ◽

Melting Curve ◽

3D Culture ◽

Gene Profiling ◽

3D Microenvironment

Abstract Assessing an optimal reference gene as an internal control for target gene normalization is important during quantitative real time polymerase chain reaction (RT-qPCR) of three-dimensional cell culture. Especially, gene profiling of cancer cells under a complex 3D microenvironment in a polymer scaffold provides a deeper understanding of recapitulation of in vivo tumors. In this aspect, expression of six housekeeping genes (HKG’s): glyceraldehyde-3-phosphodehydrogenase (GAPDH), β-actin (ACTB), beta-2-microglobulin (B2M), 18S ribosomal RNA (18S rRNA), peptidyl-propyl-isomerase A (PPIA), and ribosomal protein L13 (RPL-13)) during the monolayer culture (two-dimensional), and alginate-carboxymethylcellulose scaffold based three-dimensional (3D) cell culture conditioned up to 21 days was analyzed for hepatocellular carcinoma (Huh-7) cell line. The real-time gene expression using RT-qPCR of HCC spheroids in 3D culture were analyzed by determining the primer efficiency, melting curve and quantification cycle analysis of the selected candidate HKG’s. Further, RT-qPCR data were validated using analysis softwares i.e., geNorm and NormFinder for statistical significance. The study indicated RPL-13, 18S rRNA and B2M to be stable among selected referral HKG candidates and considered them as potential internal controls during varying cell culture conditions.

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