We show how some concentration inequalities for sampling without replacement can be used for bounding future samples. This process can be extended to bound the sum of future samples from multiple populations, and we analyse an illustrative sample allocation problem.

To achieve efficient utilization of low-concentration mine gas, reduce resource waste, and alleviate environmental pollution, high-temperature oxidation of low-concentration gas at a concentration range of 1.00% to 1.50% that is directly discharged into the atmosphere during coal mine production was oxidized to recover heat for reuse. The gas oxidation equipment was improved for the heating process, and the safety of low-concentration gas oxidation under high-temperature environment was evaluated. Experimental results showed that the reactor could provide a 1000 ℃ high-temperature oxidation environment for gas oxidation after installing high-temperature resistant ceramics. The pressure variation curves of the reactor with air and different concentrations of gas were similar. Due to the thermal expansion, the air pressure slightly increased and then returned to normal pressure. In contrast, the low-concentration gas exhibited a stable pressure response in the high-temperature environment of 1000 ℃. The outlet pressure was significantly greater than the inlet pressure, and the pressure difference between the inlet and outlet exhibited a trend to increase with the gas concentration. The explosion limit varied with the temperature and the blend with oxidation products. The ratio of measured gas pressure to air pressure after oxidation was below the explosion criterion, indicating that the measured concentration gas is still safe after the shift of explosion limit, which provides a safe concentration range for efficient use of low-concentration gas in the future.

Easy to use satellite-based water quality visualizations are needed for monitoring and understanding coastal and inland waters, but to date, no publicly accessible real-time global visualization system was in place. Here we introduce the Ulyssys Water Quality Viewer (UWQV), a Sentinel Hub EO Browser Custom script designed for qualitative views of aquatic chlorophyll and suspended sediment concentrations. The viewer avoids unmixing of the chlorophyll and suspended sediment spectral signal by visualizing these parameters together, with high concentrations of suspended sediment obscuring chlorophyll if present. Cloud masking uses the Hollstein and Braaten algorithms (existing EO Browser custom script code), additionally water surfaces are masked using the Normalized Differential Water Index. Chlorophyll is estimated using reflectance line height-based indicators such as fluorescence line height and maximum chlorophyll index. Suspended sediment is visualized based on single-band reflectances at 620 or 700 nm. Data sources are Sentinel-2 and Sentinel-3 images, allowing either 20 m spatial resolution or up to daily imaging. This visualization system is easy to operate and interpret, and combined with the data service capacity of the Sentinel Hub, it is expected that UWQV will contribute to monitoring of remote water bodies and to our overall understanding of physical limnology and aquatic ecology.

Increased salinity caused by saltwater intrusion or runoff from de-icing salts can severely affect freshwater vegetation and deteriorate aquatic ecosystems. These habitats can be restored with freshwater ecotypes (locally adapted populations) that tolerate above-normal salinity. Vallisneria americana is a prominent species in many freshwater ecosystems that responds differently to abiotic conditions such as substrate composition and fertility, so in this study we evaluated the effects of salt stress on 24 ecotypes of V. americana. Instant Ocean aquarium salt was used to create saline solutions [0.2 to 20.0 parts per thousand (ppt)], then plants were abruptly exposed to these solutions and maintained in these concentrations for 5 weeks before being visually assessed for quality and destructively harvested. Analysis of variance and non-linear regression were used to calculate LC₅₀ values – the lethal concentration of salt that reduced plant biomass and quality by 50% compared to control treatment. Growth rate and visual quality varied significantly among ecotypes, and ecotypes that were most and least sensitive to salt had 50% biomass reductions at 0.47 and 9.10 ppt, respectively. All ecotypes survived 10.0 ppt salinity concentration but none survived at 20.0 ppt, which suggests the maximum salinity concentration tolerated by these ecotypes is between 15.0 and 20.0 ppt.

Sehezar area is located in southern Tonokabon in Mazandaran province, north of Iran, near the Tarom – Hashdtjin belt. The existence of granitoid masses in the region can be important in terms of the potential of mineralization. Geochemical anomaly separation from the background is one of the important steps in mineral exploration. In the past decades, geochemical anomalies have been identified by means of various methods. Some of these separation methods include: statistical analysis methods (like univariate, bivariate, multivariate statistics), spatial statistical methods and fractal and multi-fractal methods. To identify the anomalous area, 71 stream sediment samples were collected from the area and analyzed by the ICP-MS method, and then interpreted. Initially, data were normalized and afterwards, univariate analysis (threshold limit and screening (P.N) methods) was used, in which results of the probable and definite anomaly of the threshold method were confirmed by the P.N screening method. Finally, the maps of the anomal zones were drawn. Then, bivariate analysis (Pearson correlation coefficients) and multivariate analysis on normal data were performed on SPSS software, in which factor analysis and cluster analysis were used for multivariate analysis. As a result of using the factor analysis method, six factors were identified and factor maps were drawn by the Surfer software. Also, by using cluster analysis, the variables were divided into two groups. In order for a better separation of the geochemical anomaly from the background, in addition to the threshold method, the Concentration - Area fractal method was used. Here, the fractal geometry using full-logarithmic graphs of the Concentration - Area obtained is capable of separating the stairs of different sections (background, threshold, and anomaly) with respect to the angle coefficient of the Concentration - Area plot. Then, in conclusion, results of these methods were compared and investigated, and finally, the anomalies area maps of the Au, Ag, Cu, Fe, W elements were drawn by Concentration - Area fractal and threshold methods and anomalous zones were introduced.

We derive a concentration inequality for the uncertainty in the mean computed by stratified random sampling, and provide an online sampling method based on this inequality. Our concentration inequality is versatile and considers a range of factors including: the data ranges, weights, sizes of the strata, the number of samples taken, the estimated sample variances, and whether strata are sampled with or without replacement. Sequentially choosing samples to minimize this inequality leads to a online method for choosing samples from a stratified population. We evaluate and compare the effectiveness of our method against others for synthetic data sets, and also in approximating the Shapley value of cooperative games. Results show that our method is competitive with the performance of Neyman sampling with perfect variance information, even without having prior information on strata variances. We also provide a multidimensional extension of our inequality and discuss future applications.

Immersion in salt-lake solution was adopted to periodically test the concrete chloride-ion diffusion coefficient. The regression analysis was also completed. Also investigated was the time-dependent law of concrete chloride-ion diffusion coefficient with time. The influence of chloride-ion concentration in solution, water-to-cement ratio, and corrosion time on the largeness and accumulation rate of the concrete chloride-ion diffusion coefficient was also analyzed. Test results show that the concrete chloride-ion diffusion coefficient gradually decreased with increasing time and increased with increasing chloride-ion concentration in a salt lake . Taking into account the influence of factors such as water–binder ratio, chloride-ion concentration, and time-varying characteristics, a multi-factor calculation model for the concrete chloride-ion diffusion coefficient was established. Combining the prediction results and the measured data reported in this paper, the effectiveness and applicability of the established concrete chloride-ion diffusion coefficient calculation model were compared and verified, and the durability design and service life of a concrete structure under cool chlorine were compared. The results of analysis provide important boundary conditions.

The article presents the results of tests of minor head losses through PVC and PP elbows for a flow of water and mixtures of water and sand with grain sizes of up to 0.5 mm and concentrations of 5.6 g∙L-1, 10.84 g∙L-1, and 15.73 g∙L-1. The tests were carried out at variable flow velocities for three elbow diameters of 63, 75, and 90 mm. The flow rate, pressure difference in the tested cross-sections, and temperature of the fluids were measured and automatically recorded. The results of the measurements were used to develop mathematical models for determining the minor head loss coefficient as a function of elbow diameter, sand concentration in the liquid, and Reynolds number. The mathematical model was developed by cross validation. It was shown that when the concentration of sand in the liquid was increased by 1.0 g∙L-1, the coefficient of minor head loss through the elbows increased, in the Reynolds number range of 4.6∙104 − 2.1∙105, by 0.3−0.01% for PP63, 0.6−0.03 % for PP75, 1.1−0.06 % for PP90, 0.8−0.01 % for PVC63, 0.8−0.02 % for PVC75, and 0.9−0.04 % for PVC90. An increase in Re from 5∙104 to 2∙106 for elbows with diameters of 63, 75 and 90 mm caused a 7.3 %, 6.8 %, and 6.0 % decrease in the minor head loss coefficient, respectively.

MEMThe PdNi film hydrogen sensors with Wheatstone bridge structure were designed and fabricated by the micro-electro-mechanical system (MEMS) technology. The integrated sensors consisted of four PdNi alloy film resistors. The interval two of them were shielded with silicon nitride film and used as reference resistance, while the others were used for hydrogen sensing. The PdNi alloy films and SiN films were deposited by magnetron sputtering. The morphology and microstructure of the PdNi films were characterized with X-ray diffraction (XRD). The output resistance signal was converted to millivolt output voltage signal for easy data acquisition. Hydrogen (H2) sensing properties of PdNi film hydrogen sensor with Wheatstone bridge structure was investigated under different temperatures (30℃, 50℃ and 70℃) and H2 concentrations (from 10 ppm to 0.4%). The hydrogen sensor demonstrated good response at different hydrogen concentrations and high repeatability in cycle testing under 0.4% H2 concentration. Under 10ppm hydrogen, the PdNi film hydrogen sensor had evident and collectable output voltage of 600 μV.

The water-rich liquid layer immobilized on the surface of the polar stationary phases is critical to the retention of polar compounds in hydrophilic interaction chromatography (HILIC). Although the presence of the adsorbed water layer has been investigated and confirmed by multiple techniques, there is a lack of quantitative measure that can be easily determined and linked to chromatographic parameters. This study proposes a simple measure termed volume ratio (the ratio of the adsorbed water layer volume and the mobile phase volume) that provides a relative, but quantitative information on the adsorbed water layer and may be linked to the phase ratio. The volume ratio can be easily determined using toluene elution volume. The volume ratio values are measured in 25 polar stationary phases in various mobile phase conditions. In addition to the acetonitrile content in the mobile phase, ammonium acetate concentration in the mobile phase and column temperature also have significant influences on the volume ratio and the adsorbed water layer.

Particulate matter is a severe source of atmospheric pollution in urban cities, and it has adverse effects on human health. This study was conducted during the whole year of 2016 to monitor the concentrations of PM₁₀ and PM_2.5 on the Beijing Hanshiqiao wetland and bare land in Beijing to analyze their correlations with meteorological factors and compare the removal efficiency between two land surface types. The results indicated that (1) the PM₁₀ and PM_2.5 concentrations on the bare land were higher than those on wetland as a whole, reaching the highest value both at night and dusk and the lowest value near noon. The average concentration of PM₁₀ was higher in winter (wetland: 137.48 μg·m^-3; bare land: 164.75 μg·m^-3) and spring (wetland: 205.18 μg·m^-3; bare land: 244.85 μg·m^-3) and the concentration of PM_2.5 on the wetland also reached the higher value in winter and spring with the average of 84.52 μg·m^-3 and 98.98 μg·m^-3, whereas, it was higher in spring and summer on the bare land; (2) concentrations of PM₁₀ and PM_2.5 were significantly positively affected by the relative humidity (P < 0.01) and negatively influenced by wind speed (P < 0.05). The relationship between PM₁₀ and PM_2.5 concentrations and temperature was found complicated: it showed a significantly negative correlation (P < 0.01) in winter and spring and was insignificant in autumn, but in summer, only the correlation between the PM₁₀ concentration and temperature on wetland was significant (P < 0.01); (3) the removal efficiencies of PM₁₀ and PM_2.5 followed the order of spring > winter > autumn > summer on the wetland, and the removal efficiency of PM₁₀ was greater than that of PM_2.5. This study is aim to provide practical measures to improve the air quality and facilitate sustainable development in Beijing.

This work introduced the potential synergistic toxicity of binary mixtures of pesticides and pharmaceuticals, which have been substantially detected in major river basins in South Korea. Different dose-response curve functions were employed in each experimental toxicity dataset for Aliivibrio fischeri. We tested the toxicity of 30 binary mixtures at two effect concentrations: high effect concentration [EC₅₀] and low effect concentration [EC₁₀] ranges. Thus, the toxicological interactions were evaluated at 60 effected concentration data points in total and based on model deviation ratios (MDRs) between predicted and observed toxicity values (e.g., three types of combined effects: synergistic (MDR > 2), additive (0.5 ≤ MDR ≤ 2), and antagonistic (MDR < 0.5)). From the 60 data points, MDRs could not be applied to 17 points, since their toxicities could not be measured. The result showed 48 %-additive (n = 20), 40 %-antagonistic (n = 17), and 12 %-synergistic (n = 6) toxicity effects from 43 binaries (excluding the 17 combinations without MDRs). In this study, EC₁₀ ratio mixtures at a low overall effect range showed a general tendency to have more synergistic effects than the EC₅₀ ratio mixtures at a high effect range. We also found an inversion phenomenon, which detected three binaries of the combination of synergism at low concentrations and additive antagonism at high concentrations.

Three-dimensional (3D) cell culture is considered more clinically relevant in mimicking the structural and physiological conditions of tumors in vivo compared to two-dimensional cell cultures. In recent years, high-throughput screening (HTS) in 3D cell arrays has been extensively used for drug discovery because of its usability and applicability. Herein, we developed a microfluidic spheroid culture device (μFSCD) with a concentration gradient generator (CGG) that enabled cells to form spheroids and grow in the presence of cancer drug gradients. The device is composed of concave microwells with several serpentine micro-channels which generate a concentration gradient. Once the colon cancer cells (HCT116) formed a single spheroid (approximately 120 μm in diameter) in each microwell, spheroids were perfused in the presence of the cancer drug gradient irinotecan for 3 days. The number of spheroids, roundness, and cell viability, were inversely proportional to the drug concentration. These results suggest that the μFSCD with a CGG has the potential to become an HTS platform for screening the efficacy of cancer drugs.

Background and Aim: Zinc plays a pivotal role in various zinc enzymes, resulting in the maintenance of liver function. Patients with chronic liver diseases (CLDs) usually have lower concentrations of zinc, which decrease further as liver fibrosis progresses. It remains unknown whether long-term zinc supplementation improves liver function and reduces the risk of hepatocellular carcinoma (HCC) development. Patients and Methods: Two hundred sixty-seven patients with CLDs who received a zinc preparation (Zn-group; 196 patients), or who did not receive zinc (no Zn-treatment group; 71 patients) were retrospectively analyzed in this study. The Zn-group was divided into 4 groups according to their serum Zn concentrations at 6 months after the start of Zn treatment. Results: Liver function significantly deteriorated in the no Zn-treatment group, while no notable change was observed in the Zn-group. The cumulative incidence rates of events and HCC at 3 years were lower in the Zn-group (9.5%, 7.6%) than in the no Zn-treatment group (24.9%, 19.2%) (p<0.001). According to the serum Zn concentrations, the cumulative incidence rates of events and HCC were significantly decreased in patients with Zn concentrations ≥ 70 µg/dl (p<0.001). Conclusion: Zinc supplementation appears to be effective at maintaining liver function and suppressing events and HCC development, especially among patients whose Zn concentration is greater than 70 µg/dl.

Normally, the Nernst voltage calculated from the concentration of the reaction gas in the flow channel is considered to be the ideal voltage (reversible voltage) of the oxyhydrogen fuel cell, but actually it will cause a concentration gradient when the reaction gas flows from the flow channel through the gas diffusion layer to the catalyst layer. The Nernst voltage loss in fuel cells in most of the current literature is thought to be due to the difference in concentration of reaction gas in the flow channel and concentration of reaction gas on the catalyst layer at the time when the high net current density is generated. Based on the Butler-Volmer equation in oxyhydrogen fuel cell, this paper demonstrates that the Nernst voltage loss is caused by the concentration difference of reaction gas in flow channel and on the catalytic layer at the time when equilibrium potential (Galvanic potential) of each electrode is generated.

Stingless bee beekeeping provides new opportunities to improve the incomes of many households in Malaysia through the sale of honey and other bee products. While Heterotrigona itama is one of the most commonly cultured species of stingless bees, its behavior is not very well understood. Hence, we conducted this study to investigate the behavior of H. itama in exploiting food sources by ascertaining the nectar sugar concentration preferred by the bee. We also aimed to determine the preferred distance of food source from the bee hive. Our results suggest that H. itama prefers high sugar concentrations of 35% and above, and they would fly up to 7 m from the hive to collect food. We discuss how nectar concentration and food distance influence the number of bees exploiting food sources and the overall foraging pattern of H. itama.

Gold nanoparticles (AuNPs) are currently under intense investigation for biomedical and biotechnology applications, thanks to their ease in preparation, stability, biocompatibility, multiple surface functionalities and size-dependent optical properties. The most commonly used method for AuNPs synthesis in aqueous solution is the reduction of tetrachloroauric acid (HAuCl₄) with trisodium citrate. We observed variations in the pH and concentration of the gold colloidal suspension synthesized under standard conditions, verifying a reduction in the reaction yield by around 46% from pH 5.3 (2.4 nM) to pH 4.7 (1.29 nM). Citrate-capped AuNPs were characterized by UV-visible spectroscopy, TEM, EDS and zeta-potential measurements, revealing a linear correlation between pH and the concentration of the generated AuNPs. This result can be attributed to the adverse effect of protons both on citrate oxidation and on citrate adsorption onto the gold surface, which is required to form the stabilization layer. Overall, this study provides insight into the effect of the pH over the synthesis performance of the method, which would be of particular interest from the point of view of large-scale manufacturing processes.

The sustainability of irrigated agriculture depends on the quality of irrigation water used. The electrolyte concentration (EC) of irrigation water may lead to the accumulation of salts in the root zone layers and affect the physiological functions of the crop by osmotic and ion toxicity effects. Further, the cationic and anionic composition of the water may alter the exchangeable cation composition of the soil and as well as its pH. Because of the dominance of sodium salts in many sources of irrigation water, parameters related to sodium such as exchangeable sodium percentage (ESP) of soils and sodium adsorption ratio (SAR) of soil solutions have been commonly used to study the effects of sodium in irrigation water on soil structural stability. Quirk and Schofield concept of ‘threshold electrolyte concentration’ (TEC) has shown the importance of electrolytes in preventing the effects of sodium on soil structure. Based on this concept, several models have been proposed to relate ESP or SAR with EC to predict the possible impacts of irrigation water on soil structural stability. However, many research reports indicate that this relationship varies with soils and a given model is not suitable for all types of soils. Further, the effects of potassium and magnesium in the processes leading to clay dispersion are disregarded in these models. This essay analyses all the factors involved in the structural failure of soils with different cationic composition, identify the defects in these TEC models and re-defines TEC on the basis of new insights on dispersive and flocculating charges of soils. This review does not deal with EC effects on crops and also the role of contaminant ions not involved with soil structural stability.

Influence of water concentration on the properties of fluorine-doped tin oxide (FTO) thin films was systematically studied in this work. Home made SISOM’s SPEED techniques and its chemical formulation was employed to grow the FTO on quartz substrate. Water concentration in the precursor solution was varied from 0, 0.5, 1.0 and 1.5 mol %. The structural, optical, and electrical properties of the films were studied under these deposition conditions. The results show that the properties of the films varied significantly with water concentration. Scanning electron microscopy (SEM) revealed FTO films whose grain size and uniformity increases significantly with increase in water concentration. The structure of the films was measured by X-ray diffraction (XRD) measurement. It shows polycrystalline films with (110), (101), (200), (211) and (220) orientation; the strength increases as water concentration increases. The optical transmission was determined by UV-Vis spectroscopy at 380–780 nm UV-VIS regions. The optical transmittance varies with water concentration with an average of 84%. The electrical property, measured by Hall Effect revealed n-type semiconductor. The films have the following properties: resistivity, 15 × 10^-4Ω cm; carrier concentration, 18.7 × 10¹⁹ cm^-3 and mobility of 21.86 cm² V^-1 s^-1. The average figure of merit, φ of the FTO film is 1.25. Optimum deposition condition was established after series of experiments and was found to be 1.5% water concentration at 460^oC substrate temperature. The FTO films deposited in this work could be a promising replacement to indium tin oxide (ITO) especially in dye-sensitized solar cells.

Thermal plasma technique is becoming prominent in the treatment of variety of waste ranging from municipal solid waste, incinerator residue, hospital waste, electronics waste and industrial sludge. Application of the new treatment technology to petroleum sludge requires information on the nature and characteristics of the sludge that will be use to optimize the treatment system. In this investigation, petroleum sludge obtained from Petronas Melaka was characterized for its physical and chemical features. Proximate and ultimate analysis as well as determination of elemental composition were carried out. The sludge was found to contain high moisture (78.91%), low ash (5.06%), low volatiles (5.52%) and high fixed carbon (10.51%). The sludge has a TOC of 54.48% and HHV of 23.599MJ/kg. Despite the high moisture content, the higher heating value (HHV) is high when compared to literature values. The high value of HHV may be associated with the high fixed carbon, low ash content and high value of TOC. The apparent density of the sludge is slightly lower. Fourteen heavy metals are detected in significant quantities. Proper waste management that will safely dispose the sludge is required. The waste disposal technique should take into cognizant the possibility of leaching of heavy metals into ground water on one hand and the gasification of lighter ones on the other.

Background: Globally, hot cooked refined rice is consumed in large quantities and is a major contributor to dietary glycaemic load. This study aimed to compare the glycaemic potency of hot and cold stored parboiled rice to widely available medium grain white rice. Method: Twenty-eight healthy volunteers participated in a three treatment experiment where postprandial blood glucose was measured over 120 minutes after consumption of 140g of rice. Three rice samples were freshly cooked medium-grain white rice, freshly cooked parboiled rice, and parboiled rice stored overnight at 4 ºC. All rice was served warm at 65 ºC. Chewing time was recorded. Results: The 24-hours cold-stored and reheated parboiled rice resulted in a 42% and 12% lower blood glucose concentration trajectory than freshly cooked medium-grain white rice and freshly cooked parboiled rice. Chewing time for 10g cold stored parboiled rice was 6 seconds (25%) longer and was considered more palatable, visually appealing and better tasting than freshly cooked medium grain (all P<0.05). Conclusions:. For regular consumers of rice reheating cooked rice after cold storage would lower the dietary glycaemic load and long term may reduce risk for type 2 and gestational diabetes. More trials are needed to identify the significance.

To obtain smooth coatings of TiO2 for building a new design of Ti-6Al-4V heart valve, the anodic oxidation technique in pre-spark conditions was evaluated. TiO2 coating is necessary for its recognize biocompatibility and corrosion resistance. A required feature on surfaces in contact with blood is a low level of roughness (Ra ≤ 50 nm) that not favor the formation of blood clots. The present paper compares the coatings obtained by anodic oxidation of the Ti-6Al-4V alloy using H2SO4 at different concentrations (0.1 M to 4 M) as electrolyte and applying different voltages (from 20 V to 70 V). Color and morphological analysis of coatings are performed using optical and scanning microscopy. The crystalline phases were analyzed by glancing X-ray diffraction. By varying the applied voltage different interference colors coatings were obtained. The differences in morphologies of the coatings, due to the change in concentration, are more evident at high voltages limiting the oxidation conditions for the desired application. Anatase phase was detected from 70 V for 1 M H2SO4. An increase in the concentration of H2SO4 decreases the voltage at which the transformation of amorphous to crystalline coatings occurs, i.e. with 4 M H2SO4 the anatase phase appears at 60 V.

The paper presents a method to assess the effect of electrolyte additives on the energy capacity of Pb-acid batteries. The method applies to any chemically unreactive additive, including suspensions and gels. The approach is thermodynamically based and it leads to the definition of a region of admissible concentrations –the battery’s admissible range– where the battery can undergo an indefinite number of charge/discharge cycles without suffering permanent damage. An experimental procedure to determine this range is presented. The obtained results provide a way to assess the potential of electrolyte additives to improve the energy capacity of Pb-acid batteries. The same results also provide a means to determine the additive concentration that produces the maximum energy capacity increase of the battery. The paper closes with an example of application of the proposed approach to a practical case.

This paper proposed three-dimensional numerical simulation method by coupling of electrostatic and fluid fields to evaluating the performance of electrical sensor in the concentration measurement of gas/solid two-phase flow. Compared with the static numerical simulation, this real-time dynamic 3-D simulation method can research on a designed capacitance sensor combining the dynamic characteristics of the two-phase flows for concentration measurement. Several fluid-electrostatic models of transmission pipes with different sensor structures are built. Under different test positions and different particle concentrations, the flow characteristics and the corresponding electric signals can be obtained, and the correlation coefficient between the concentration values and the capacitance values are used for performance evaluation of the sensors. The effects of flow regimes on concentration measurement are also been investigated in this paper. To validate the results of simulation, an experimental platform with horizontal straight pipe for phase volume concentration measurement of solid/air two-phase flow is built, and the experimental results agree well with simulation conclusions. The simulation and test results show that the coupling models can give constructive reference opinions for the sensor design and collection of installation position in different transmission pipelines, which are very important for the practical process of pneumatic conveying system.

Acellular bacterial ghosts (BGs) are empty non-living bacterial cell envelopes, commonly generated by controlled expression of the cloned lysis gene E of bacteriophage PhiX174. In this study, Vibrio parahaemolyticus ghosts (VPGs) were generated by chemically induced lysis and the method is based on minimum inhibitory concentration (MIC) of sodium hydroxide (NaOH), acetic acid, boric acid, citric acid, maleic acid, hydrochloric acid and sulfuric acid. The MIC values of the respective chemicals were 3.125, 6.25, < 50.0, 25.0, 6.25, 1.56 and 0.781 mg/ml. Except boric acid, the lysis efficiency was reached more than 99.99% at 5 min after treatment of all chemicals. Among those chemicals, NaOH-induced VPGs showed completely DNA-free that was confirmed by quantitative real-time PCR. Besides, lipopolysaccharides (LPS) extracted from the NaOH-induced VPGs showed no distinctive band on SDS-PAGE gel after silver staining. On the other hand, LPS extracted from wild-type bacterial cells as well as the organic acids-induced VPGs showed triple major bands and LPS extracted from the inorganic acids-induced VPGs showed double bands. It suggests that some surface structures in LPS of the NaOH-induced VPGs may be lost, weakened or modified by the MIC of NaOH. Nevertheless, Limulus amoebocyte lysate assay revealed that there is no significant difference in endotoxic activity between the NaOH-induced VPGs and wild-type bacterial cells. Macrophages exposed to the NaOH-induced VPGs at 0.5 × 106 CFU/mL showed cell viability of 97.9%, however the MIC of NaOH did not reduce the cytotoxic effect of wild-type bacterial cells. Like Escherichia coli LPS, the NaOH-induced VPGs are an excellent activator of pro-inflammatory cytokines (IL-1β and iNOS), anti-inflammatory cytokine (IL-10) and dual activities (IL-6) in the stimulated macrophage cells. On the other hand, the induction of TNF-α mRNA was remarkable in the macrophages exposed with wild-type cells. Scanning electron microscopy showed the formation of trans-membrane lysis tunnel structures in the NaOH-induced VPGs. SDS-PAGE and agarose gel electrophoresis also confirmed that cytoplasmic proteins and genomic DNA released from the VPGs to culture medium through the lysis tunnel structures. Taken together, all these results indicated that the NaOH-induced VPGs show the potency of safe, economical and effective inactivated bacterial vaccine candidate.