Comparison of the limulus amebocyte lysate test and gas chromatography-mass spectrometry for measuring lipopolysaccharides (endotoxins) in airborne dust from poultry-processing industries

The lipopolysaccharide (endotoxin) content in airborne dust samples from three different poultry slaughterhouses was determined with both the chromogenic Limulus amebocyte lysate assay and gas chromatography-mass spectrometry analysis of lipopolysaccharide-derived 3-hydroxy fatty acids. Gram-negative cell walls were also measured by using two-dimensional gas chromatography/electron-capture analysis of diaminopimelic acid originating from the peptidoglycan. The correlation between the results of the Limulus assay and those of gas chromatography-mass spectrometry for determination of the lipopolysaccharide content in the dust samples was poor, whereas a good correlation was obtained between lipopolysaccharide and diaminopimelic acid concentrations with the gas chromatographic methods. The results suggest that it is predominantly cell-wall-dissociated lipopolysaccharides that are measured with the Limulus assay, whereas the gas chromatographic methods allow determination of total concentrations of lipopolysaccharide, including Limulus-inactive lipopolysaccharide, gram-negative cells, and cellular debris.     

Comparison of the limulus amebocyte lysate test and gas chromatography-mass spectrometry for measuring lipopolysaccharides (endotoxins) in airborne dust from poultry-processing industries.

The lipopolysaccharide (endotoxin) content in airborne dust samples from three different poultry slaughterhouses was determined with both the chromogenic Limulus amebocyte lysate assay and gas chromatography-mass spectrometry analysis of lipopolysaccharide-derived 3-hydroxy fatty acids. Gram-negative cell walls were also measured by using two-dimensional gas chromatography/electron-capture analysis of diaminopimelic acid originating from the peptidoglycan. The correlation between the results of the Limulus assay and those of gas chromatography-mass spectrometry for determination of the lipopolysaccharide content in the dust samples was poor, whereas a good correlation was obtained between lipopolysaccharide and diaminopimelic acid concentrations with the gas chromatographic methods. The results suggest that it is predominantly cell-wall-dissociated lipopolysaccharides that are measured with the Limulus assay, whereas the gas chromatographic methods allow determination of total concentrations of lipopolysaccharide, including Limulus-inactive lipopolysaccharide, gram-negative cells, and cellular debris.     

Ab initio DFT study of structural and mechanical properties of methane and carbon dioxide hydrates

The structural and mechanical properties of methane and carbon dioxide hydrates were investigated using density functional theory simulations. Well-established equations of state of solids and exchange-correlation functionals were used for fitting the unit lattice total energy as a function of volume, and the full second-order elastic constants of these two gas hydrates were determined by energy–strain analyses. The polycrystalline elastic properties were also calculated from the unit lattice results. The final results for methane hydrate agree well with available experimental data and with other theoretical results. The two gas hydrates were found to be highly elastically isotropic, but they differed significantly in shear properties. The presented results for carbon dioxide hydrates are the first complete set reported so far. The results are a significant contribution to the ab initio material characterisation of gas hydrates required for ongoing fundamental studies and technological applications.     

A method of reconstruction of clinical gas-analyzer signals corrupted by positive-pressure ventilation.

The use of sidestream infrared and paramagnetic clinical gas analyzers is widespread in anesthesiology and respiratory medicine. For most clinical applications, these instruments are entirely satisfactory. However, their ability to measure breath-by-breath volumetric gas fluxes, as required for measurement of airway dead space, oxygen uptake, and so on, is usually inferior to that of the mass spectrometer, and this is thought to be due, in part, to their slower response times. We describe how volumetric gas analysis with the Datex Ultima analyzer, although reasonably accurate for spontaneous ventilation, gives very inaccurate results in conditions of positive-pressure ventilation. We show that this problem is a property of the gas sampling system rather than the technique of gas analysis itself. We examine the source of this error and describe how cyclic changes in airway pressure result in variations in the flow rate of the gas within the sampling catheter. This results in the phenomenon of "time distortion," and the resultant gas concentration signal becomes a nonlinear time series. This corrupted signal cannot be aligned or integrated with the measured flow signal. We describe a method to correct for this effect. With the use of this method, measurements required for breath-by-breath gas-exchange models can be made easily and reliably in the clinical setting.     

Comparison of protein fermentation characteristics in rumen fluid determined with the gas production technique and the nylon bag technique

In this study, a modified version of the gas production technique was used to determine protein fermentation characteristics in rumen fluid of 19 feedstuffs. Performing the incubations in a N-free environment, and with an excess of rapidly fermentable carbohydrates, made N the limiting factor to microbial growth, and so gas production profiles reflected the availability of N from the feed samples. Results showed that fermentation of protein in rumen fluid can be determined with this modified gas production technique, and that there were distinct differences in protein fermentation between the feed samples. Availability of protein for fermentation was highest in wheat, potato pieces and lupin, and lowest in Rumiraap, a formaldehyde treated rapeseed meal, palm kernel expeller and brewery grains. The protein degradation characteristics of the 19 feed ingredients were also determined with the in situ nylon bag technique. With the obtained results, the amount of rumen escape protein (REP) was calculated for each feedstuff. The results showed that the rate of degradation ranged from 0.010/h for Rumiraap to 0.151/h for wheat. The amount of REP ranged from 197 g/kg CP for lupin to 840 g/kg CP for Rumiraap. Comparing the gas production results with the results obtained with the nylon bag technique showed that there was a good relationship between the gas production after 12–25 h of incubation and the calculated amount of REP (r² = 0.83–0.85). The results show that the adapted gas production technique, being depleted of N and using an excess of rapidly fermentable carbohydrates, is suitable to recognize differences in N availability between feed samples and can be used as an alternative to the nylon bag technique and other in vitro techniques.     

Use of cyanobacterial gas vesicles as oxygen carriers in cell culture

The gas vesicles isolated from the cells of filamentous cyanobacterium Anabaena flos-aquae were treated and sterilized with glutaraldehyde and then evaluated for their effectiveness as gas carriers in cell culture. Anchorage-dependent Vero cells were grown in a packed bed of microcarrier beads under the perfusion of Dulbecco’s Modified Eagle’s Medium with 1% serum. The culture medium supplemented with 1.8% (v/v) gas vesicles was found to support a 30% higher maximum glucose utilization rate than the same medium without gas vesicles. The gas vesicle suspension was confirmed to have no apparent effects on cell metabolism in T-flask cultures. The study results indicated that the gas vesicles, with high oxygen carrying capacity, can be used to increase the oxygen supply in cell culture systems.     

Development of a gas chromatographic selected ion monitoring assay for thyroxine (T4) in human serum

The components of a gas chromatographic mass spectrometric-selected ion monitoring (SIM) assay for thyroxine (T4) in human serum are described. The internal standard for the assay was synthesized from deuterium-labelled 3,5-diiodotyrosine and 3,5-diiodo-4-hydroxyphenylpyruvic acid. A novel method was developed for isolating the products of the coupling reaction. The results obtained by gas chromatography mass spectrometry SIM were compared with those of radioimmunoassay. The gas chromatographic mass spectrometric SIM assay would form the basis of a reference assay for T4.     

Insights into large-scale cell-culture reactors: II. Gas-phase mixing and CO₂ stripping

Most discussions about stirred tank bioreactors for cell cultures focus on liquid-phase motions and neglect the importance of the gas phase for mixing, power input and especially CO(2) stripping. Particularly in large production reactors, CO(2) removal from the culture is known to be a major problem. Here, we show that stripping is mainly affected by the change of the gas composition during the movement of the gas phase through the bioreactor from the sparger system towards the headspace. A mathematical model for CO(2)-stripping and O(2)-mass transfer is presented taking gas-residence times into account. The gas phase is not moving through the reactor in form of a plug flow as often assumed. The model is validated by measurement data. Further measurement results are presented that show how the gas is partly recirculated by the impellers, thus increasing the gas-residence time. The gas-residence times can be measured easily with stimulus-response techniques. The results offer further insights on the gas-residence time distributions in stirred tank reactors.     

NS0 cell damage by high gas velocity sparging in protein-free and cholesterol-free cultures

Recent developments in high cell density and high productivity fed-batch animal cell cultures have placed a high demand on oxygenation and carbon dioxide removal in bioreactors. The high oxygen demand is often met by increasing agitation and sparging rates of air/O2 in the bioreactors. However, as we demonstrate in this study, an increase of gas sparging can result in cell damage at the sparger site due to high gas entrance velocities. Previous studies have showed that gas bubble breakup at the culture surface was primarily responsible for cell damage in sparged bioreactors. Such cell damage can be reduced by use of surfactants such as Pluronic F-68 in the culture. In our results, where NS0 cells were grown in a protein-free and cholesterol-free medium containing 0.5 g/L Pluronic F-68, high gas entrance velocity at the sparger site was observed as the second mechanism for cell damage. Experiments were performed in scaled-down spinners to model the effect of hydrodynamic force resulting from high gas velocities on antibody-producing NS0 cells. Cell growth and cell death were described by first-order kinetics. Cell death rate constant increased significantly from 0.04 to 0.18 day(-1) with increasing gas entrance velocity from 2.3 to 82.9 m/s at the sparger site. The critical gas entrance velocity for the NS0 cell line studied was found to be approximately 30 m/s; velocities greater than 30 m/s caused cell damage which resulted in reduced viability and consequently reduced antibody production. Observations from a second cholesterol-independent NS0 cell line confirmed the occurrence of cell damage due to high gas velocities. Increasing the concentration of Pluronic F-68 from 0.5 to 2 g/L had no additional protective effect on cell damage associated with high gas velocity at the sparger. The results of gas velocity analysis for cell damage have been applied in two case studies of large-scale antibody manufacturing. The first is a troubleshooting study for antibody production carried out in a 600 L bioreactor, and the second is the development of a gas sparger design for a large bioreactor scale (e.g., 10,000 L) for antibody manufacturing.     

Modeling of Gas Production from Shale Reservoirs Considering Multiple Transport Mechanisms

Gas transport in unconventional shale strata is a multi-mechanism-coupling process that is different from the process observed in conventional reservoirs. In micro fractures which are inborn or induced by hydraulic stimulation, viscous flow dominates. And gas surface diffusion and gas desorption should be further considered in organic nano pores. Also, the Klinkenberg effect should be considered when dealing with the gas transport problem. In addition, following two factors can play significant roles under certain circumstances but have not received enough attention in previous models. During pressure depletion, gas viscosity will change with Knudsen number; and pore radius will increase when the adsorption gas desorbs from the pore wall. In this paper, a comprehensive mathematical model that incorporates all known mechanisms for simulating gas flow in shale strata is presented. The objective of this study was to provide a more accurate reservoir model for simulation based on the flow mechanisms in the pore scale and formation geometry. Complex mechanisms, including viscous flow, Knudsen diffusion, slip flow, and desorption, are optionally integrated into different continua in the model. Sensitivity analysis was conducted to evaluate the effect of different mechanisms on the gas production. The results showed that adsorption and gas viscosity change will have a great impact on gas production. Ignoring one of following scenarios, such as adsorption, gas permeability change, gas viscosity change, or pore radius change, will underestimate gas production.     

Time series analysis models for estimation of greenhouse gas emitted by different sectors in Turkey

Due to the increasing global warming in the world, analyzing greenhouse gas emissions is a crucial issue. This study has examined greenhouse gas emissions in Turkey according to energy sector, industrial processes sector, agriculture sector and waste sector. Then, time series analysis models are used to estimate greenhouse gas emissions based on sectors. Models' performances are tested using mean error, mean absolute error and root mean square error. The results show that forecasting models have a good potential to estimate the national greenhouse gas emissions for different sector within a reasonable error. The study results will help organize and estimate the national greenhouse gas emissions inventory.     

Ion drift in a gas in an external electric field

A model of ion collisions with gas atoms is constructed that takes into account resonant ion-atom charge exchange, polarization interaction, and short-range (gas-kinetic) collisions. The cross sections for resonant charge exchange of the ions of noble gases, as well as rubidium, cesium, and mercury ions, with atoms are calculated based on the experimental data and the results of numerical simulations of ion collisions with parent gas atoms in a uniform electric field. The effect of the electric field, the temperature of the buffer gas atoms, and the percentage composition of the gas mixture on the ion velocity distribution function is investigated.     

Life Cycle Inventories for the Nuclear and Natural Gas Energy Systems,and Examples of Uncertainty Analysis (14 pp)

Goal, Scope and Background The energy systems included in the ecoinvent database v1.1 describe the situation around year 2000 of Swiss and Western European power plants and boilers with the associated energy chains. The addressed nuclear systems concern Light Water Reactors (LWR) with mix of open and closed fuel cycles. The system model ‘Natural Gas’ describes production, distribution, and combustion of natural gas. Methods Comprehensive life cycle inventories of the energy systems were established and cumulative results calculated within the ecoinvent framework. Swiss conditions for the nuclear cycle were extrapolated to major nuclear countries. Long-term radon emissions from uranium mill tailings have been estimated with a simplified model. Average natural gas power plants were analysed for different countries considering specific import/export of the gas, with seven production regions separately assessed. Uncertainties have been estimated quantitatively. Results and Discussion Different radioactive emission species and wastes are produced from different steps of the nuclear cycle. Emissions of greenhouse gases from the nuclear cycle are mostly from the upstream chain, and the total is small and decreasing with increasing share of centrifuge enrichment. The results for natural gas show the importance of transport and low pressure distribution network for the methane emissions, whereas energy is mostly invested for production and long-distance pipeline transportation. Because of significant differences in power plant efficiencies and gas supply, country specific averages differ greatly. Conclusion The inventory describes average worldwide supply of nuclear fuel and average nuclear reactors in Western Europe. Although the model for nuclear waste management was extrapolated from Swiss conditions, the ranges obtained for cumulative results can represent the average in Europe. Emissions per kWh electricity are distributed very differently over the natural gas chain for different species. Modern combined cycle plants show better performance for several burdens like cumulative greenhouse gas emissions compared to average plants. Recommendation and Perspective Comparison of country-specific LWRs or LWR types on the basis of these results is not recommended. Specific issues on different strategies for the nuclear fuel cycle or location-specific characteristics would require extension of analysis. Results of the gas chain should not be directly applied to areas other than those modelled because emission factors and energy requirements may differ significantly. A future update of inventory data should reconsider production and transport from Russia, as it is a major producer and exporter to Europe. The calculated ranges of uncertainty factors in ecoinvent provide useful information but they are more indications of uncertainties rather than strict 95% intervals, and should therefore be applied carefully.     

An experimental study on biomass air-steam gasification in a fluidized bed

The characteristics of biomass air-steam gasification in a fluidized bed are studied in this paper. A series of experiments have been performed to investigate the effects of reactor temperature, steam to biomass ratio (S/B), equivalence ratio (ER) and biomass particle size on gas composition, gas yield, steam decomposition, low heating value (LHV) and carbon conversion efficiency. Over the ranges of the experimental conditions used, the fuel gas yield varied between 1.43 and 2.57 Nm3/kg biomass and the LHV of the fuel gas was between 6741 and 9143 kJ/Nm3. The results showed that higher temperature contributed to more hydrogen production, but too high a temperature lowered gas heating value. The LHV of fuel gas decreased with ER. Compared with biomass air gasification, the introduction of steam improved gas quality. However, excessive steam would lower gasification temperature and so degrade fuel gas quality. It was also shown that a smaller particle was more favorable for higher gas LHV and yield.     

Genetic transformation of a halophilic archaebacterium with a gas vesicle gene cluster restores its ability to float.

The halophilic archaebacterium, Halobacterium halobium, and many other aquatic bacteria synthesize gas-filled vesicles for flotation. We recently identified a cluster of 13 genes (gvpMLKJIHGFEDACN) on a 200-kb H. halobium plasmid, pNRC100, involved in gas vesicle synthesis. We have cloned and reconstructed the gvp gene cluster on an H. halobium-E. coli shuttle plasmid. Transformation of H. halobium Vac- mutants lacking the entire gas vesicle gene region with the gvp gene cluster results in restoration of their ability to float. These results open the way toward further genetic analysis of gas vesicle gene functions and directed flotation of other microorganisms with potential biotechnological applications.     

呼吸气体一次进样气相色谱分析方法

本文报道在 SP-2305E 型气相色谱仪上,采用由聚合物固定相 GDX-104柱(80/100目,φ4×600ram)、钠石灰柱(40/60目,φ4×350mm)及5A 分子筛柱(60/80目,φ4×2000mm)所组成的单气路双柱跨鉴定器(热导池)色谱流程,以氢为载气(40ml/min),桥流160mA,柱箱温度50°С,一次进样测定呼吸气体氧及二氧化碳含量的方法。进样量1ml,完成一次分析的周期为4 1/2min。氧测定的精确度与 Scholander 微量呼吸气体分析方法相当,二氧化碳测定的精确度则优于后者。还比较了分别以氢或氩为载气时的定量结果。并就定量方法及“氩校正”问题进行了讨论。     

Ab initio conformational study of N-acetyl-L-proline-N',N'-dimethylamide: a model for polyproline

We report here the results on N-acetyl-l-proline-N',N'-dimethylamide (Ac-Pro-NMe2) as a model for polyproline at the HF/6-31+G(d) level with the conductor-like polarizable continuum model of self-consistent reaction field methods to figure out the conformational preference and cis-trans isomerization of polyproline in the gas phase, chloroform, methanol, and water. The second methyl substitution at the carboxyl amide end results in different backbone structures and their populations from those of N-acetyl-L-proline-N-methylamide (Ac-Pro-NHMe). In particular, all conformations with the C7 hydrogen bond between acetyl and amide ends, which is the most probable conformations of Ac-Pro-NHMe in the gas phase and in nonpolar solvents, disappeared for Ac-Pro-NMe2 even in the gas phase due to the lack of amide hydrogen. The dominant conformation for Ac-Pro-NMe2 is the polyproline II structure with the trans prolyl peptide bond in the gas phase and in solutions. In methanol, the population of the polyproline I structure with the cis prolyl peptide bond is calculated to be larger than that in water, which is consistent with experiments. It should be noted that Ac-Pro-NMe2 has higher rotational barriers for the cis-trans isomerization of the Ac-Pro peptide bond than Ac-Pro-NHMe in the gas phase and in solutions, which could be due to the lack of the intramolecular hydrogen bond between prolyl nitrogen and carboxyl N-H group for the transition state of Ac-Pro-NMe2. The rotational barriers for Ac-Pro-NMe2 are increased with the increase of solvent polarity, as seen for Ac-Pro-NHMe.     

Molecular simulation of size-selective gas adsorption in idealised carbon nanotubes

Molecular simulations were used to examine the adsorption of diatomic molecules (nitrogen and oxygen) and similarly sized gases (argon and methane) in pores with van der Waals diameters similar in size to the gas diameters. Idealised carbon nanotubes were used to model generic pores, to better understand the effect of pore diameter on guest adsorption in the absence of defects, specific adsorption sites, or variations in pore diameter that often complicate studies of gas adsorption in other porous materials. Molecular dynamics simulations of open nanotubes show that argon and methane are able to enter tubes whose diameters are slightly smaller than the gas diameters. Diatomic gases are able to enter tubes that are significantly smaller than their kinetic diameters with the molecular axis aligned parallel to the nanotube. The results indicate that size-selective adsorption of these gases is theoretically possible, although differences in pore diameters of only a few tenths of an Angstrom are required. Grand canonical Monte Carlo simulations of a 3.38 Å nanotube indicate significant uptake by argon and oxygen, but not nitrogen or methane. The adsorption of nitrogen and methane gradually increases as the nanotube diameter approaches 4.07 Å, and all gases fully saturate a 4.54 Å nanotube. Of the nanotubes studied, the largest adsorption enthalpy for any gas corresponds to the 4.54 Å nanotube, with significantly lower enthalpies seen in the 5.07 Å nanotube. These results suggest an ideal pore diameter for each gas based on the gas–pore van der Waals interaction energies. Trends in the ideal diameter correlate with the minimum tube diameter accessible to each gas.     

A model evaluation of estimates of breath-to-breath alveolar gas exchange

A mathematical model has been implemented for evaluation of methods for estimating breath-to-breath alveolar gas exchange during exercise in humans. This model includes a homogeneous alveolar gas exchange compartment, a dead space compartment, and tissue spaces for CO2 (alveolar and dead space). The dead space compartment includes a mixing portion surrounded by tissue and an unmixed (slug flow) portion which is partitioned between anatomical and apparatus contributions. A random sinusoidal flow pattern generates a breath-to-breath variation in pulmonary stores. The Auchincloss algorithm for estimating alveolar gas exchange (Auchincloss et al., J. Appl. Physiol. 21: 810-818, 1966) was applied to the model, and the results were compared with the simulated gas exchange. This comparison indicates that a compensation for changes in pulmonary stores must include factors for alveolar gas concentration change as well as alveolar volume change and thus implies the use of end-tidal measurements. Although this algorithm yields reasonable estimates of breath-to-breath alveolar gas exchange, it does not yield a "true" indirect measurement because of inherent error in the estimation of a homogeneous alveolar gas concentration at the end of expiration.