Optimization of Lead Bioremediation by Marine Halomonas sp. ES015 Using Statistical Experimental Methods

Bioremediation technology is now used for treatment instead of traditional metal removal methods. A strain was isolated from Marsa Alam, Red sea, Egypt showed high resistance to high lead concentration and was identified by the 16S rRNA gene sequencing technique as Halomonas sp. ES015. Medium optimization was carried out using Plackett-Burman design, and the most significant factors were yeast extract, casamino acid and inoculums size. The optimized media obtained by the statistical design raised the removal efficiency from 84% to 99% from initial concentration 250 ppm of lead. Moreover, Box-Behnken experimental design was applied to study the relationship between yeast extract concentration, casamino acid concentration and inoculums size. The optimized medium increased removal efficiency to 97% from initial concentration 500 ppm of lead. Immobilized Halomonas sp. ES015 cells on sponge cubes, using optimized medium in loop bioremediation column, showed relatively constant lead removal efficiency when reused six successive cycles over the range of time interval. Also metal removal efficiency was not affected by flow rate changes. Finally, the results of this research refer to the possibility of lead bioremediation by free or immobilized cells of Halomonas sp. ES015. Also, bioremediation can be done in batch cultures and semicontinuous cultures using column technology.

The Study of Stable Isotopes (18O, 2H & 13C) in Kardeh River and Dam Reservoir, North-Eastern Iran

Among various water resources, the surface water has a dominant role in providing water supply in the arid and semi-arid region of Iran. Andarokh-Kardeh basin is located in 50 km from Mashhad city - the second biggest city of Iran (NE of Iran), draining by Kardeh river which provides a significant portion of potable and irrigation water needs for Mashhad. The stable isotopes (18O, 2H,13C-DIC, and 13C-DOC), as reliable and precious water fingerprints, have been measured in Kardeh river (Kharket, Mareshk, Jong, All and Kardeh stations) and in Kardeh dam reservoirs (at five different sites S1 to S5) during March to June 2011 and June 2012. On δ18O vs. δ2H diagram, the river samples were plotted between Global and Eastern Mediterranean Meteoric Water lines (GMWL and EMMWL) which demonstrate that various moisture sources are providing humidity for precipitation events in this area. The enriched δ18O and δ2H values (-6.5 ‰ and -44.5 ‰ VSMOW) of Kardeh dam reservoir are compared to Kardeh river (-8.6‰and-54.4‰), and its deviation from Mashhad meteoric water line (MMWL- δ2H=7.16δ18O+11.22) is due to evaporation from the open surface water body. The enriched value of δ 13C-DIC and high amount of DIC values (-7.9 ‰ VPDB and 57.23 ppm) in the river and Kardeh dam reservoir (-7.3 ‰ VPDB and 55.53 ppm) is due to dissolution of Mozdooran Carbonate Formation lithology (Jm1 to Jm3 units) (contains enriched δ13C DIC values of 9.2‰ to 27.7‰ VPDB) in the region. Because of the domination of C3 vegetations in Andarokh_Kardeh basin, the δ13C-DOC isotope of the river (-28.4‰ VPDB) and dam reservoir (-32.3‰ VPDB) demonstrate depleted values. Higher DOC concentration in dam reservoir (2.57 ppm) compared to the river (0.72 ppm) is due to more biologogical activities and organic matters in dam reservoir.

Total and Leachable Concentration of Trace Elements in Soil towards Human Health Risk, Related with Coal Mine in Jorong, South Kalimantan, Indonesia

Coal mining is well known to cause considerable environmental impacts, including trace element contamination of soil. This study aimed to assess the trace element (As, Cd, Co, Cu, Ni, Pb, Sb, and Zn) contamination of soil in the vicinity of coal mining activities, using the case study of Asam-asam River basin, South Kalimantan, Indonesia, and to assess the human health risk, incorporating total and bioavailable (water-leachable and acid-leachable) concentrations. The results show the enrichment of As and Co in soil, surpassing the background soil value. Contamination was evaluated based on the index of geo-accumulation, Igeo and the pollution index, PI. Igeo values showed that the soil was generally uncontaminated (Igeo ≤ 0), except for elevated As and Co. Mean PI for Ni and Cu indicated slight contamination. Regarding the assessment of health risks, the Hazard Index, HI showed adverse risks (HI > 1) for Ni, Co, and As. Further, Ni and As were found to pose unacceptable carcinogenic risk (risk > 1.10-5). Farming, settlement, and plantation were found to present greater risk than coal mines. These results show that coal mining activity in the study area contaminates the soils by particular elements and may pose potential human health risk in its surrounding area. This study is important for setting appropriate countermeasure actions and improving basic coal mining management in Indonesia.

Effects of Temperature and Enzyme Concentration on Quality of Pineapple and Pawpaw Blended Juice

The effects of temperature and enzyme concentration on the quality of mixed pineapple and pawpaw blended fruits juice were studied. Extracts of the two fruit juices were separately treated at 70  for 15 min each so as to inactivate micro-organisms. They were analyzed and blended in different proportions of 70% pawpaw and 30% pineapple, 60% pawpaw and 40% pineapple, 50% pineapple and 50% pawpaw, 40% pawpaw and 60% pineapple. The characterization of the fresh pawpaw and pineapple juice before blending showed that the juices have good quality. The high water content of the product may have affected the viscosity, vitamin C content and total soluble solid of the blended juice to be low. The effects of the process parameters on the quality showed that better quality of the blended juice can be obtained within the optimum temperature range of (50-70 °C) and enzyme concentration range (0.12-0.18 w/v). The ratio of mix 60% pineapple juice: 40% pawpaw juice has better quality. This showed that pawpaw and pineapple juices can blend effectively to produce a quality juice.

Impacts of Climate Change on Water Resources of Greater Zab and Lesser Zab Basins, Iraq, Using Soil and Water Assessment Tool Model

The Greater Zab and Lesser Zab are the major tributaries of Tigris River contributing the largest flow volumes into the river. The impacts of climate change on water resources in these basins have not been well addressed. To gain a better understanding of the effects of climate change on water resources of the study area in near future (2049-2069) as well as in distant future (2080-2099), Soil and Water Assessment Tool (SWAT) was applied. The model was first calibrated for the period from 1979 to 2004 to test its suitability in describing the hydrological processes in the basins. The SWAT model showed a good performance in simulating streamflow. The calibrated model was then used to evaluate the impacts of climate change on water resources. Six general circulation models (GCMs) from phase five of the Coupled Model Intercomparison Project (CMIP5) under three Representative Concentration Pathways (RCPs) RCP 2.6, RCP 4.5, and RCP 8.5 for periods of 2049-2069 and 2080-2099 were used to project the climate change impacts on these basins. The results demonstrated a significant decline in water resources availability in the future.

Heavy Metals Estimation in Coastal Areas Using Remote Sensing, Field Sampling and Classical and Robust Statistic

Sediments are an important source of accumulation of toxic contaminants within the aquatic environment. Bioassays are a powerful tool for the study of sediments in relation to their toxicity, but they can be expensive. This article presents a methodology to estimate the main physical property of intertidal sediments in coastal zones: heavy metals concentration. This study, which was developed in the Bay of Santander (Spain), applies classical and robust statistic to CASI-2 hyperspectral images to estimate heavy metals presence and ecotoxicity (TOC). Simultaneous fieldwork (radiometric and chemical sampling) allowed an appropriate atmospheric correction to CASI-2 images.

Honey Contamination in the Republic of Kazakhstan

This study involves detailed information about contaminants of honey in the Republic of Kazakhstan. The requirements of the technical regulation ‘Requirements to safety of honey and bee products’ and GOST 19792-2001 were taken into account in this research. Contamination of honey by antibiotics wqs determined by the IEA (immune-enzyme analysis), Ridder analyzer and Tecna produced test systems. Voltammetry (TaLab device) was used to define contamination by salts of heavy metals and gamma-beta spectrometry, ‘Progress BG’ system, with preliminary ashing of the sample of honey was used to define radioactive contamination. This article pointed out that residues of chloramphenicol were detected in 24% of investigated products, in 22% of them –streptomycin, in 7.3% - sulfanilamide, in 2.4% - tylosin, and in 12% - combined contamination was noted. Geographically, the greatest degree of contamination of honey with antibiotics occurs in the Northern Kazakhstan – 54.4%, and Southern Kazakhstan - 50%, and the lowest in Central and Eastern Kazakhstan with 30% and 25%, respectively. Generally, pollution by heavy metals is within acceptable limits, but the contamination from lead is highest in the Akmola region. The level of radioactive cesium and strontium is also within acceptable concentrations. The highest radioactivity in terms of cesium was observed in the East Kazakhstan region - 49.00±10 Bq/kg, in Akmola, North Kazakhstan and Almaty - 12.00±5, 11.05±3 and 19.0±8 Bq/kg, respectively, while the norm is 100 Bq/kg. In terms of strontium, the radioactivity in the East Kazakhstan region is 25.03±15 Bq/kg, while in Akmola, North Kazakhstan and Almaty regions it is 12.00±3, 10.2±4 and 1.0±2 Bq/kg, respectively, with the norm of 80 Bq/kg. This accumulation is mainly associated with the environmental degradation, feeding and treating of bees. Moreover, in the process of collecting nectar, external substances can penetrate honey. Overall, this research determines factors and reasons of honey contamination.

Modeling of Electrokinetic Mixing in Lab on Chip Microfluidic Devices

This paper sets to demonstrate a modeling of electrokinetic mixing employing electroosmotic stationary and time-dependent microchannel using alternate zeta patches on the lower surface of the micromixer in a lab on chip microfluidic device. Electroosmotic flow is amplified using different 2D and 3D model designs with alternate and geometric zeta potential values such as 25, 50, and 100 mV, respectively, to achieve high concentration mixing in the electrokinetically-driven microfluidic system. The enhancement of electrokinetic mixing is studied using Finite Element Modeling, and simulation workflow is accomplished with defined integral steps. It can be observed that the presence of alternate zeta patches can help inducing microvortex flows inside the channel, which in turn can improve mixing efficiency. Fluid flow and concentration fields are simulated by solving Navier-Stokes equation (implying Helmholtz-Smoluchowski slip velocity boundary condition) and Convection-Diffusion equation. The effect of the magnitude of zeta potential, the number of alternate zeta patches, etc. are analysed thoroughly. 2D simulation reveals that there is a cumulative increase in concentration mixing, whereas 3D simulation differs slightly with low zeta potential as that of the 2D model within the T-shaped micromixer for concentration 1 mol/m3 and 0 mol/m3, respectively. Moreover, 2D model results were compared with those of 3D to indicate the importance of the 3D model in a microfluidic design process.

Flow Performance of Hybrid Cement Based Mortars

The workability of hybrid alkaline cements is a field of knowledge that still needs further research efforts. This paper reports experimental results of 32 hybrid cement mixes regarding the joint effect of sodium hydroxide concentration, the use of a commercial superplasticizer and a biopolymer on the flow and compressive strength performance. The results show that the use of commercial admixtures led to a slightly increase in the flow of mortars with lower sodium hydroxide concentration.

Unsteady Natural Convection Heat and Mass Transfer of Non-Newtonian Casson Fluid along a Vertical Wavy Surface

Detailed numerical calculations are illustrated in our investigation for unsteady natural convection heat and mass transfer of non-Newtonian Casson fluid along a vertical wavy surface. The surface of the plate is kept at a constant temperature and uniform concentration. To transform the complex wavy surface to a flat plate, a simple coordinate transformation is employed. The resulting partial differential equations are solved using the fully implicit finite difference method with SUR procedure. Flow and heat transfer characteristics are investigated for a wide range of values of the Casson parameter, the dimensionless time parameter, the buoyancy ratio and the amplitude-wavelength parameter. It is found that, the variations of the Casson parameter have significant effects on the fluid motion, heat and mass transfer. Also, the maximum and minimum values of the local Nusselt and Sherwood numbers increase by increase either the Casson parameter or the buoyancy ratio.

Analysis and Design of Inductive Power Transfer Systems for Automotive Battery Charging Applications

Transferring electrical power without any wiring has been a dream since late 19th century. There were some advances in this area as to know more about microwave systems. However, this subject has recently become very attractive due to their practiScal systems. There are low power applications such as charging the batteries of contactless tooth brushes or implanted devices, and higher power applications such as charging the batteries of electrical automobiles or buses. In the first group of applications operating frequencies are in microwave range while the frequency is lower in high power applications. In the latter, the concept is also called inductive power transfer. The aim of the paper is to have an overview of the inductive power transfer for electrical vehicles with a special concentration on coil design and power converter simulation for static charging. Coil design is very important for an efficient and safe power transfer. Coil design is one of the most critical tasks. Power converters are used in both side of the system. The converter on the primary side is used to generate a high frequency voltage to excite the primary coil. The purpose of the converter in the secondary is to rectify the voltage transferred from the primary to charge the battery. In this paper, an inductive power transfer system is studied. Inductive power transfer is a promising technology with several possible applications. Operation principles of these systems are explained, and components of the system are described. Finally, a single phase 2 kW system was simulated and results were presented. The work presented in this paper is just an introduction to the concept. A reformed compensation network based on traditional inductor-capacitor-inductor (LCL) topology is proposed to realize robust reaction to large coupling variation that is common in dynamic wireless charging application. In the future, this type compensation should be studied. Also, comparison of different compensation topologies should be done for the same power level.

Optimization of the Co-Precipitation of Industrial Waste Metals in a Continuous Reactor System

A continuous copper precipitation treatment (CCPT) system was conceived at Intel Chandler Site to serve as a first-of-kind (FOK) facility-scale waste copper (Cu), nickel (Ni), and manganese (Mn) co-precipitation facility. The process was designed to treat highly variable wastewater discharged from a substrate packaging research factory. The paper discusses metals co-precipitation induced by internal changes for manufacturing facilities that lack the capacity for hardware expansion due to real estate restrictions, aggressive schedules, or budgetary constraints. Herein, operating parameters such as pH and oxidation reduction potential (ORP) were examined to analyze the ability of the CCPT System to immobilize various waste metals. Additionally, influential factors such as influent concentrations and retention times were investigated to quantify the environmental variability against system performance. A total of 2,027 samples were analyzed and statistically evaluated to measure the performance of CCPT that was internally retrofitted for Mn abatement to meet environmental regulations. In order to enhance the consistency of the influent, a separate holding tank was cannibalized from another system to collect and slow-feed the segregated Mn wastewater from the factory into CCPT. As a result, the baseline influent Mn decreased from 17.2+18.7 mg1L-1 at pre-pilot to 5.15+8.11 mg1L-1 post-pilot (70.1% reduction). Likewise, the pre-trial and post-trial average influent Cu values to CCPT were 52.0+54.6 mg1L-1 and 33.9+12.7 mg1L-1, respectively (34.8% reduction). However, the raw Ni content of 0.97+0.39 mg1L-1 at pre-pilot increased to 1.06+0.17 mg1L-1 at post-pilot. The average Mn output declined from 10.9+11.7 mg1L-1 at pre-pilot to 0.44+1.33 mg1L-1 at post-pilot (96.0% reduction) as a result of the pH and ORP operating setpoint changes. In similar fashion, the output Cu quality improved from 1.60+5.38 mg1L-1 to 0.55+1.02 mg1L-1 (65.6% reduction) while the Ni output sustained a 50% enhancement during the pilot study (0.22+0.19 mg1L-1 reduced to 0.11+0.06 mg1L-1). pH and ORP were shown to be significantly instrumental to the precipitative versatility of the CCPT System.

Improvement of Antibacterial Activity for Ceftazidime by Partially Purified Tannase from Penicillium expansum

Tannase has wide applications in food, beverage, brewing, cosmetics and chemical industries and one of the major applications of tannase is the production of gallic acid. Gallic acid is used for manufacturing of trimethoprim. In the present study, a local fungal strain of Penicillium expansum A4 isolated from spoilt apple samples gave the highest production level of tannase. Tannase was partially purified with a recovery yield of 92.52% and 6.32 fold of purification by precipitation using ammonium sulfate at 50% saturation. Tannase led to increased antimicrobial activity of ceftazidime against Pseudomonas aeruginosa and S. aureus and had a synergism effect at low concentrations of ceftazidime, and thus, tannase may be a useful adjuvant agent for the treatment of many bacterial infections in combination with ceftazidime.

Flexural Strength of Alkali Resistant Glass Textile Reinforced Concrete Beam with Prestressing

Due to the aging of bridges, increasing of maintenance costs and decreasing of structural safety is occurred. The steel corrosion of reinforced concrete bridge is the most common problem and this phenomenon is accelerating due to abnormal weather and increasing CO2 concentration due to climate change. To solve these problems, composite members using textile have been studied. A textile reinforced concrete can reduce carbon emissions by reduced concrete and without steel bars, so a lot of structural behavior studies are needed. Therefore, in this study, textile reinforced concrete beam was made and flexural test was performed. Also, the change of flexural strength according to the prestressing was conducted. As a result, flexural strength of TRC with prestressing was increased compared and flexural behavior was shown as reinforced concrete.

Human Health Risk Assessment from Metals Present in a Soil Contaminated by Crude Oil

The main sources of soil pollution due to petroleum contaminants are industrial processes involve crude oil. Soil polluted with crude oil is toxic for plants, animals, and humans. Human exposure to the contaminated soil occurs through different exposure pathways: Soil ingestion, diet, inhalation, and dermal contact. The present study research is focused on soil contamination with heavy metals as a consequence of soil pollution with petroleum products. Human exposure pathways considered are: Accidentally ingestion of contaminated soil and dermal contact. The purpose of the paper is to identify the human health risk (carcinogenic risk) from soil contaminated with heavy metals. The human exposure and risk were evaluated for five contaminants of concern of the eleven which were identified in soil. Two soil samples were collected from a bioremediation platform from Muntenia Region of Romania. The soil deposited on the bioremediation platform was contaminated through extraction and oil processing. For the research work, two average soil samples from two different plots were analyzed: The first one was slightly contaminated with petroleum products (Total Petroleum Hydrocarbons (TPH) in soil was 1420 mg/kgd.w.), while the second one was highly contaminated (TPH in soil was 24306 mg/kgd.w.). In order to evaluate risks posed by heavy metals due soil pollution with petroleum products, five metals known as carcinogenic were investigated: Arsenic (As), Cadmium (Cd), ChromiumVI (CrVI), Nickel (Ni), and Lead (Pb). Results of the chemical analysis performed on samples collected from the contaminated soil evidence soil contamination with heavy metals as following: As in Site 1 = 6.96 mg/kgd.w; As in Site 2 = 11.62 mg/kgd.w, Cd in Site 1 = 0.9 mg/kgd.w; Cd in Site 2 = 1 mg/kgd.w; CrVI was 0.1 mg/kgd.w for both sites; Ni in Site 1 = 37.00 mg/kgd.w; Ni in Site 2 = 42.46 mg/kgd.w; Pb in Site 1 = 34.67 mg/kgd.w; Pb in Site 2 = 120.44 mg/kgd.w. The concentrations for these metals exceed the normal values established in the Romanian regulation, but are smaller than the alert level for a less sensitive use of soil (industrial). Although, the concentrations do not exceed the thresholds, the next step was to assess the human health risk posed by soil contamination with these heavy metals. Results for risk were compared with the acceptable one (10-6, according to World Human Organization). As, expected, the highest risk was identified for the soil with a higher degree of contamination: Individual Risk (IR) was 1.11×10-5 compared with 8.61×10-6. 

Removal of Total Petroleum Hydrocarbons from Contaminated Soils by Electrochemical Method

Soil contamination phenomena are a wide world issue that has received the important attention in the last decades. The main pollutants that have affected soils are especially those resulted from the oil extraction, transport and processing. This paper presents results obtained in the framework of a research project focused on the management of contaminated sites with petroleum products/ REMPET. One of the specific objectives of the REMPET project was to assess the electrochemical treatment (improved with polarity change respect to the typical approach) as a treatment option for the remediation of total petroleum hydrocarbons (TPHs) from contaminated soils. Petroleum hydrocarbon compounds attach to soil components and are difficult to remove and degrade. Electrochemical treatment is a physicochemical treatment that has gained acceptance as an alternative method, for the remediation of organic contaminated soils comparing with the traditional methods as bioremediation and chemical oxidation. This type of treatment need short time and have high removal efficiency, being usually applied in heterogeneous soils with low permeability. During the experimental tests, the following parameters were monitored: pH, redox potential, humidity, current intensity, energy consumption. The electrochemical method was applied in an experimental setup with the next dimensions: 450 mm x 150 mm x 150 mm (L x l x h). The setup length was devised in three electrochemical cells that were connected at two power supplies. The power supplies configuration was provided in such manner that each cell has a cathode and an anode without overlapping. The initial value of TPH concentration in soil was of 1420.28 mg/kgdw. The remediation method has been applied for only 21 days, when it was already noticed an average removal efficiency of 31 %, with better results in the anode area respect to the cathode one (33% respect to 27%). The energy consumption registered after the development of the experiment was 10.6 kWh for exterior power supply and 16.1 kWh for the interior one. Taking into account that at national level, the most used methods for soil remediation are bioremediation (which needs too much time to be implemented and depends on many factors) and thermal desorption (which involves high costs in order to be implemented), the study of electrochemical treatment will give an alternative to these two methods (and their limitations).

Gasification of Trans-4-Hydroxycinnamic Acid with Ethanol at Elevated Temperatures

Lignin is a major constituent of woody biomass, and exists abundantly in nature. It is the major byproducts from the paper industry and bioethanol production processes. The byproducts are mainly used for low-valued applications. Instead, lignin can be converted into higher-valued gaseous fuel, thereby helping to curtail the ever-growing price of oil and to slow down the trend of global warming. Although biochemical treatment is capable of converting cellulose into liquid ethanol fuel, it cannot be applied to the conversion of lignin. Alternatively, it is possible to convert lignin into gaseous fuel thermochemically. In the present work, trans-4-hydroxycinnamic acid, a model compound for lignin, which closely resembles the basic building blocks of lignin, is gasified in an autoclave with ethanol at elevated temperatures and pressures, that are above the critical point of ethanol. Ethanol, instead of water, is chosen, because ethanol dissolves trans-4-hydroxycinnamic acid easily and helps to convert it into lighter gaseous species relatively well. The major operating parameters for the gasification reaction include temperature (673-873 K), reaction pressure (5-25 MPa) and feed concentration (0.05-0.3 M). Generally, more than 80% of the reactant, including trans-4-hydroxycinnamic acid and ethanol, were converted into gaseous products at an operating condition of 873 K and 5 MPa.

Study on Bending Characteristics of Square Tube Using Energy Absorption Part

In the square tube subjected to the bending load, the rigidity of the entire square tube is reduced when a collapse occurs due to local stress concentration. Therefore, in this research, the influence of bending load on the square tube with attached energy absorbing part was examined and reported. The analysis was conducted by using Finite Element Method (FEM) to produced bending deflection and buckling points. Energy absorption was compared from rigidity of attached part and square tube body. Buckling point was influenced by the rigidity of attached part and the thickness rate of square tube.

Economic Optimization of Shell and Tube Heat Exchanger Using Nanofluid

Economic optimization of shell and tube heat exchanger (STHE) is presented in this paper. To increase the rate of heat transfer, copper oxide (CuO) nanoparticle is added into the tube side fluid and their optimum results are compared with the case of without additive nanoparticle. Total annual cost (TAC) is selected as fitness function and nine decision variables related to the heat exchanger parameters as well as concentration of nanoparticle are considered. Optimization results reveal the noticeable improvement in the TAC and in the case of heat exchanger working with nanofluid compared with the case of base fluid (8.9%). Comparison of the results between two studied cases also reveal that the lower tube diameter, tube number, and baffle spacing are needed in the case of heat exchanger working with nanofluid compared with the case of base fluid.

Cytotoxic Effect of Crude Extract of Sea Pen Virgularia gustaviana on HeLa and MDA-MB-231 Cancer Cell Lines

Marine organisms such as soft coral, sponge, ascidians, and tunicate containing rich source of natural compound have been studied in last decades because of their special chemical compounds with anticancer properties. The aim of this study was to investigate anti-cancer property of ethyl acetate extracted from marine sea pen Virgularia gustaviana found from Persian Gulf coastal (Bandar Abbas). The extraction processes were carried out with ethyl acetate for five days. Thin layer chromatography (TLC) and high-performance liquid chromatography (HPLC) were used for qualitative identification of crude extract. The viability of HeLa and MDA-Mb-231 cancer cells was investigated using MTT assay at the concentration of 25, 50, and a 100 µl/ml of ethyl acetate is extracted. The crude extract of Virgularia gustaviana demonstrated ten fractions with different Retention factor (Rf) by TLC and Retention time (Rt) evaluated by HPLC. The crude extract dose-dependently decreased cancer cell viability compared to control group. According to the results, the ethyl acetate extracted from Virgularia gustaviana inhibits the growth of cancer cells, an effect which needs to be further investigated in the future studies.