Investigation of Reactive Oxygen Species production in Human Hepatocytes

1. Aim/Background: Reactive oxygen species (ROS) have been identified as compounds responsible for producing cellular damage. The purpose of this research is to examine if there is production of reactive oxygen species through free radical intermediates within human hepatocytes treated with morphine, bilirubin, or furosemide. The investigation examines the early stages of biotransformation by measuring the levels of reactive oxygen species produced inside of the treated hepatocytes within the first and second hours of treatment. The experiment was designed upon a case of a jaundiced (elevated bilirubin) infant who received morphine and furosemide and later died through unknown mechanisms. The experiment looks to examine if these drug compounds could contribute to cellular damage. This can help to further understand the potential interactions and complications of free radical intermediates produced during the phases of biotransformation. 2. Method: Previously cultured human hepatocytes were washed by centrifugation and re-suspended in 1x supplemental buffer to a concentration of 1x106 cells/mL and seeded in a dark clear bottom 96-well microplate at 100,000 stained cells/well. The cells were treated with either furosemide, morphine, bilirubin, a Tert-Butyl hydro peroxide (TBHP) positive control, or left as a background. Reactive oxygen generated in the presence of these agents were quantified by fluorescence excitation/emission measurement at 495nm/529nm. Fluorescence was measured at one and two hours. ROS generated convert 2',7'-dichlorodihydrofluorescein diacetate to 2',7'-dichlorodihydrofluorescein within the cells, which fluoresces. The fluorescence intensity detected is equivalent to the level of ROS generated. Wells that were untreated were used as blanks and subtracted from background and TBPH. 3. Results: Furosemide and Morphine did not produce statistically significant levels of ROS (p >0.05) above the background in both hours 1 and 2 of biotransformation and ROS measurement (Figure 1). Although Bilirubin did not produce statistically significant (p >0.05) levels of ROS above the background (Figure 2) during the first hour, it did produce statistically significant levels in the second hour of biotransformation. Each compound’s level of ROS was reduced during the second hour, signaling the removal of intermediate ROS metabolites (Figure 2). The production of ROS in each compound signifies that there is biotransformation to an intermediate that produces ROS. 4. Conclusion: The production of ROS above the background by each of the compounds shows there is an intermediate free radical compound that is produced during the biotransformation of each compound [21]. In this study, although furosemide and morphine did not produce statistically significant levels of ROS in both hours of biotransformation, bilirubin did produce significant levels of ROS in the second hour of biotransformation. This finding is in line with previous studies that shows morphine to offer protective effects against ROS production [16, 17]; and bilirubin demonstrating deleterious production of ROS at high doses [18]. Further work must be done to examine the correlation between the levels of ROS and extent of hepatocellular damage.

Verification and validation of linear gyrokinetic and kinetic-MHD simulations for internal kink instability in DIII-D tokamak

Verification and linear validation of the internal kink instability in tokamak have been performed for both gyrokinetic (GTC) and kinetic-MHD codes (GAM-solver, M3D-C1-K, NOVA, XTOR-K). Using realistic magnetic geometry and plasma profiles from the same equilibrium reconstruction of the DIII-D shot #141216, these codes exhibit excellent agreement for the growth rate and mode structure of the internal kink mode when all kinetic effects are suppressed. The simulated radial mode structures, obtained from linear simulations, are in reasonable agreement with the normalised electron cyclotron emission measurement after adjusting, within the experimental uncertainty, the safety factor q=1 flux-surface location in the equilibrium reconstruction. Compressible magnetic perturbations strongly destabilize the kink, while poloidal variations of the equilibrium current density reduce the growth rate of the kink. Furthermore, kinetic effects of thermal ions are found to decrease the kink growth rate in kinetic-MHD simulations, but increase the kink growth rate in gyrokinetic simulations, due to the additional drive of the ion temperature gradient and parallel electric field. Kinetic thermal electrons are found to have negligible effects on the internal kink instability.

On the Issues of Spatial Modeling of Non-Standard Profiles by the Example of Electromagnetic Emission Measurement Data

This paper examines the possibility of the spatial modelling of the Earth’s natural pulsed-electromagnetic-field measured values, which form a closed profile without the data inside. This geophysical method allows us to map active tectonic movement which breaches the integrity of pipes. During the experiment, 4.5 km of profiles were measured in the Admiralteysky district of St. Petersburg, Russia. Regular electromotive force (EMF) values and anomalous EMF values were obtained, ranging from 0 to 900 µV and above 900 µV, respectively. The anomalous values are associated with tectonic faults in the bedrock. The data obtained are characterized by complex spatial anisotropy associated with the development of two groups of tectonic faults of different orientations. The authors have considered the problems of the spatial modeling of the data obtained. The main problems, the solutions to which should allow the obtaining of adequate models, have been identified. Based on the analysis of the measurement results, geological features of the studied areas, as well as variography, the following possible solutions were proposed: changing the measurement technique; dividing the data array according to the main directions of anisotropy; the need to introduce additional correction coefficients. The problem revealed in this article requires further research on the basis of the obtained results, which will reduce the cost and timing of such studies, and, as a result, give an opportunity to take into account active tectonic disturbances during the construction and scheduled maintenance of underground utilities, which is especially important within the framework of the concept of sustainable development.

Study of Real-road Nitrogen Oxide Emissions of Non-road Vehicles

The exhaust gas pollutants of the non-road vehicles are harmful to the environment. Many non-road vehicles meet the requirements of the regulations in the laboratory. However, the real-road emissions of such vehicles are sometimes higher. Measuring the real-road emissions of non-road vehicles is very important. The real-road emissions are measured by on-Board Diagnostics (OBD), but there are some problems in the data stability of OBD. The NOx emissions of a bulldozer (a type of China IV non-road vehicle) based on both portable emission measurement system (PEMS) and OBD are studied in this article. Experiments contained three working processes: idle, driving, and operating. The nitrogen oxide (NOx) emissions during operating were highest. The NOx emission characteristics of the bulldozer from PEMS and OBD have the similar variation trends. But there are still some differences, including the NOx emission value and response time. The measurement principles and different sampling points between PEMS and OBD are the main factors. An effective data processing method is introduced to reduce the differences of between the data from PEMS and OBD. Briefly, the NOx emissions of the OBD and PEMS were highly consistent. The OBD is reliable and can be widely used in non-road vehicles.

The Development of CO2 Instantaneous Emission Model of Full Hybrid Vehicle with the Use of Machine Learning Techniques

Road transport contributes to almost a quarter of carbon dioxide emissions in the EU. To analyze the exhaust emissions generated by vehicle flows, it is necessary to use specialized emission models, because it is infeasible to equip all vehicles on the road in the tested road sections with the Portable Emission Measurement System (PEMS). However, the currently used emission models may be inadequate to the investigated vehicle structure or may not be accurate due to the used macroscale. This state of affairs is especially related to full hybrid vehicles, since there are none of the microscale emission models that give estimated emissions values exclusively for this kind of drive system. Several automakers over the past decade have invested in hybrid vehicles with great opportunities to reduce costs through better design, learning, and economies of scale. In this work, the authors propose a methodology for creating a CO2 emission model, which takes relatively little computational time, and the models created give viable results for full hybrid vehicles. The creation of an emission model is based on the review of the accuracy results of methods, such as linear, robust regression, fine, medium, coarse tree, linear, cubic support vector machine (SVM), bagged trees, Gaussian process regression (GPR), and neural network (NNET). Particularly in the work, the best fit for the road input data for the CO2 emission model creation was the GPR method. PEMS data was used, as well as model training data and model validation. The model resulting from this methodology can be used for the analysis of emissions from simulation tests, or they can be used for input parameters for speed, acceleration, and road gradient.

Research on Computer Assisted Intelligent Emission System of High-rise Building Based on BIM Technology and Wireless Automatic Monitoring

In response to the needs of low-carbon design and carbon emission measurement of public buildings, we use advanced information tools, building information model BIM technology to solve these problems, and from the perspective of strategic development, we will establish a set of home improvement design systems based on BIM technology for unification Three-dimensional home improvement design. The system includes analysis of thermal performance of high-rise building decoration and building light environment, and proposes optimization countermeasures for low-carbon design.

The air quality index based on measurements of mobile air quality monitoring station at the waste-to-energy incineration plant PLTSa Bantargebang

Ambient air quality monitoring at waste-to-energy incineration pilot plant PLTSa Bantargebang is performed using a mobile monitoring station. The mobile monitoring station is equipped with meteorological and emission (CO, O3, NO2, PM10, PM2.5, and SO2) measurement. The monitoring was performed for 24 hour with 1 minute intervals. The emission measurement data was analyzed using Indonesian Air pollution standard index regulation (PermenLHK P.14/2020). The CO, O3, NO2, PM10, and SO2 index were in good category (1-50), while the PM2.5 index was classified as moderate (65.992). The results show that the air quality at PLTSa Bantargebang is still acceptable for human health.

Development of a Low-Cost Exhaust H2 Measurement Method for In-Use Vehicles

Port-injected hydrogen (H2) can be used as a partial substitution of diesel fuel in compression-ignition engines to reduce GHG emissions. For port-injected H2 systems, incomplete combustion or valve overlap can result in H2 slip, which increases the brake-specific fuel consumption. In this study, a low-cost method is developed to measure the H2 slip in the exhaust of a heavy-duty truck under real-world operating conditions. The truck is equipped with a 2016 15L Detroit diesel engine converted to run in dual-fuel mode with port-injected H2 ignited by directly injected diesel. Existing H2 detecting methods used for steady-state laboratory tests either have slow response time or require well-controlled testing environments. To develop a method suitable for transient on-road H2 measurements, we utilized a low-cost semiconductor sensor. The output of the sensor is potentially influenced by temperature, relative humidity (RH), gas flow rate, as well as the sensor’s resistance in the ambient air (R0) and the pre-heating strategy. Firstly, the characteristics of R0 was investigated in controlled benchtop tests, where pre-heating time, gas temperature, and RH were monitored. Then, the sensor was calibrated using a standard gas mixture of H2 and nitrogen. Finally, a Portable Emission Measurement System (PEMS) was developed to control the conditions of the sample gas. The sensor output was recorded using a low-cost Raspberry Pi Data Acquisition (DAQ) system in combination with an analog HAT (Hardware attached on top) module at a frequency of 4Hz. The results from the benchtop tests show that RH and flow rate both have significant influences on the sensor’s output. To ensure a stable R0, thirty minutes of pre-heating time is required. After calibration, the sensor’s readings are within 15% difference compared with the actual values. Data from the on-road tests demonstrated the applicability of the system for in-use vehicle’s exhaust H2 measurement. It was found from this data that the sensor’s average response time to rising H2 concentrations is 4.5s, but that the response to decreasing concentrations is much slower. The exhaust H2 concentrations, together with the engine operating data, were used to generate H2 emission maps, which provide insight into the relationship between the engine load, engine speed and the H2 slip. With further sensor development and sample gas control, this method can achieve high accuracy and extended application in in-use vehicle’s H2 emission measurements.