Accuracy and precision of protein–ligand interaction kinetics determined from chemical shift titrations

NMR-monitored chemical shift titrations for the study of weak protein?Cligand interactions represent a rich source of information regarding thermodynamic parameters such as dissociation constants (K _D ) in the micro- to millimolar range, populations for the free and ligand-bound states, and the kinetics of interconversion between states, which are typically within the fast exchange regime on the NMR timescale. We recently developed two chemical shift titration methods wherein co-variation of the total protein and ligand concentrations gives increased precision for the K _D value of a 1:1 protein?Cligand interaction (Markin and Spyracopoulos in J Biomol NMR 53: 125?C138, 2012). In this study, we demonstrate that classical line shape analysis applied to a single set of ¹H?C¹⁵N 2D HSQC NMR spectra acquired using precise protein?Cligand chemical shift titration methods we developed, produces accurate and precise kinetic parameters such as the off-rate (k _off ). For experimentally determined kinetics in the fast exchange regime on the NMR timescale, k _off ?~?3,000?s^?1 in this work, the accuracy of classical line shape analysis was determined to be better than 5?% by conducting quantum mechanical NMR simulations of the chemical shift titration methods with the magnetic resonance toolkit GAMMA. Using Monte Carlo simulations, the experimental precision for k _off from line shape analysis of NMR spectra was determined to be 13?%, in agreement with the theoretical precision of 12?% from line shape analysis of the GAMMA simulations in the presence of noise and protein concentration errors. In addition, GAMMA simulations were employed to demonstrate that line shape analysis has the potential to provide reasonably accurate and precise k _off values over a wide range, from 100 to 15,000?s^?1. The validity of line shape analysis for k _off values approaching intermediate exchange (~100?s^?1), may be facilitated by more accurate K _D measurements from NMR-monitored chemical shift titrations, for which the dependence of K _D on the chemical shift difference (????) between free and bound states is extrapolated to ?????=?0. The demonstrated accuracy and precision for k _off will be valuable for the interpretation of biological kinetics in weakly interacting protein?Cprotein networks, where a small change in the magnitude of the underlying kinetics of a given pathway may lead to large changes in the associated downstream signaling cascade.     

Release Of Petroleum Hydrocarbons From Bioremediated Soils

This article presents a qualitative evaluation of the extent to which the bioavailability (release) of a chemical is related to the biodegradation of hydrocarbons in a field bioremediation unit. The objectives of this research were to (1) quantify the rate of release of petroleum hydrocarbons from two soils that were bioremediated, (2) explore hydrocarbon release as a process affecting bioremediation; and (3) investigate the impact of bioremediation on chemical release in the two soils. An experimental protocol was used to quantify the rate of release of these hydrocarbons from two soils that had been bioremediated in a field-scale prepared bed land treatment unit. One soil showed little change in hydrocarbon concentration during 55 weeks of prepared bed bioremediation. The field study results indicated that, prior to the bioremediation, this soil had reached an environmentally acceptable endpoint. The second soil showed considerable hydrocarbon loss as a result of the bioremediation. The rate of hydrocarbon release was determined for the first soil and for the second soil at time zero and after 1, 2, and 7 months of prepared bed bioremediation. The results indicated: (1) the fraction (F) of the specific hydrocarbons that were released rapidly from the soil and the rates of release (k₂) of the residual hydrocarbons that were released slowly, (2) that the mass of each chemical of concern that was released from the first soil was very low; and (3) that the hydrocarbon released rapidly from the second soil decreased as treatment progressed. The experiments also verified, qualitatively, that some portion of each chemical evaluated was not able to be released, and thus was unavailable for bioremediation in the prepared bed land treatment unit.     

A three-time-point method for assessing kinetic parameters of 64Cu-labeled Ramucirumab trapping in VEGFR-2 positive lung tumors

ObjectiveTo describe a three-time-point method for estimating kinetic parameters involved in ⁶⁴Cu-labeled Ramucirumab (⁶⁴Cu-NOTA-RamAb) trapping of VEGFR-2 positive lung tumors.Materials and methodsPositron emission tomography (microPET) data of tumor-bearing mice for ⁶⁴Cu-NOTA-RamAb trapping in VEGFR-2 positive HCC4006 tumor were used, involving tissue activity measurements acquired at 3, 24 and 48 h post-injection, without and with administration of RamAb blocking dose. A kinetic model provided an analytical formula describing the tissue time-activity-curve, involving ⁶⁴Cu-NOTA-RamAb uptake (Ki), release rate constant (k_R) and fraction of free tracer in blood and interstitial volume (F).ResultsFitting analytical formula outcomes on mean microPET data yielded values of the kinetic parameters: Ki = 0.0314/0.0123 gram of blood per hour per gram of tissue, k_R = 0.0387/0.0313 h⁻¹ and F = 0.2075/0.2007 gram of blood per gram of tissue, without/with RamAb blocking dose, respectively (R = 0.99999 for the graph displaying microPET versus theoretical data; P < .01).ConclusionsThree independent kinetic parameters (Ki, k_R and F) can be assessed from three data points acquired at early, mid and late imaging, i.e., at 3, 24 and 48 h post-injection, for further characterization of ⁶⁴Cu-NOTA-RamAb trapping in VEGFR-2 positive lung tumors.     

Nitrogen and Phosphorus Release from Decomposing Leaves under Sub‐Humid Tropical Conditions1

For many soils of the tropics, inputs of organic materials are essential to sustain soil fertility and crop production. Research in the quality of organic inputs, a key factor controlling rates of decomposition and nutrient release, continues to guide selection and use of organic materials as nutrient sources. The relationship between decomposition patterns and the quality parameters of the fresh leaves of six agroforestry species: Sesbania sesban, Croton megalocarpus, Calliandra calothyrsus, Tithonia diversifolia, Lantana camara, and Senna spectabilis, was investigated in a litterbag study over a period of 77 days in the highlands of western Kenya. The litterbags were buried 1 cm below the soil surface and covered with soil of ca 1 cm thickness. Percent leaf mass and total N and P that remained with time strongly correlated with total P and C/P ratio (R²= 0.60‐0.90) during the first 35 days of study; but afterwards, correlation was stronger with the initial soluble polyphenolics (Pp)/P ratio (R²= 0.69‐0.92) than with total P and C/P ratio. Loss of leaf mass and release of N and P followed the exponential function, y_t= y₀* e‐^kt, from which the specific decay rate constants (k) were calculated for loss of leaf mass (k_B) and release of N (k_N) and P (K_p). Among the plant species, the k values were lowest in Calliandra with k_B= 0.012/d, k_N= 0.017/d and k_p= 0.044/d. Lantana had the highest K values with k_g= 0.067/d and k_p= 0.119/d, but the highest k_N value of 0.109/d occurred in Tithonia. The k_B values for all organic materials were lower than their corresponding k_N and k_p values, suggesting that leaching of N and P from litters may have augmented the microbial mineralization of N and P. There was a strong correlation between the k_B, k_N, and k_p values and total P (r = 0.82‐0.96; P 0.01), but not total N, lignin (LIG), or Pp. Rates of N and P release followed the general trend: Tithonia > Senna > Lantana > Sesbania > Croton > Calliandra. The results indicated that, among the quality parameters studied, total P is the most important factor controlling rate of decomposition and N and P release from organic inputs in the area of study.     

怀槐细胞悬浮培养的结构化动力学模型

为探明怀槐细胞生长、异黄酮染料木素合成与底物消耗间的关系,建立了怀槐细胞悬浮培养的结构化动力学模型。模型预测分析了胞内外的蔗糖代谢、胞内结构组分变化、胞内中间组分的变化、细胞呼吸损失以及胞内外异黄酮染料木素的合成情况。模型各参数灵敏度的分析表明kM_b1、k_b2 和k_p是最为灵敏的参数,其调节10%时,目标函数变化的最大比例分别达12.8%、4.61%和2.54%,其它参数对目标函数变化的影响均小于0.5%。该模型预测值与实验值具有较好的吻合性。     

Effect of Mineral and Organic Soil Constituents on Microbial Mineralization of Organic Compounds in a Natural Soil

This research addressed the effect of mineral and organic soil constituents on the fate of organic compounds in soils. Specifically, it sought to determine how the associations between organic chemicals and different soil constituents affect their subsequent biodegradation in soil. Four ¹⁴C-labeled surfactants were aseptically adsorbed to montmorillonite, kaolinite, illite, sand, and humic acids. These complexes were mixed with a woodlot soil, and ¹⁴CO₂ production was measured over time. The mineralization data were fitted to various production models by nonlinear regression, and a mixed (3/2)-order model was found to most accurately describe the mineralization patterns. Different mineralization patterns were observed as a function of the chemical and soil constituents. Surfactants that had been preadsorbed to sand or kaolinite usually showed similar mineralization kinetics to the control treatments, in which the surfactants were added to the soil as an aqueous solution. Surfactants that had been bound to illite or montmorillonite were typically degraded to lesser extents than the other forms, while surfactant-humic acid complexes were degraded more slowly than the other forms. The desorption coefficients (K_d) of the soil constituent-bound surfactants were negatively correlated with the initial rates of degradation (k₁) and estimates of ¹⁴CO₂ yield (P_o) as well as actual total yields of ¹⁴CO₂. However, there was no relationship between K_d and second-stage zero-order rates of mineralization (k_o). Microbial community characteristics (biomass and activity) were not correlated with any of the mineralization kinetic parameters. Overall, this study showed that environmental form had a profound effect on the ultimate fate of biodegradable chemicals in soil. This form is defined by the physicochemical characteristics of the chemical, the composition and mineralogy of the soil, and the mode of entry of the chemical into the soil environment.     

Formulation and <Emphasis Type="Italic">In Vitro</Emphasis> and <Emphasis Type="Italic">In Vivo</Emphasis> Evaluation of Lipid-Based Terbutaline Sulphate Bi-layer Tablets for Once-Daily Administration

The objective of this study was to prepare and evaluate terbutaline sulphate (TBS) bi-layer tablets for once-daily administration. The bi-layer tablets consisted of an immediate-release layer and a sustained-release layer containing 5 and 10 mg TBS, respectively. The sustained-release layer was developed by using Compritol®888 ATO, Precirol® ATO 5, stearic acid, and tristearin, separately, as slowly eroding lipid matrices. A full 4?×?2² factorial design was employed for optimization of the sustained-release layer and to explore the effect of lipid type (X ₁), drug–lipid ratio (X ₂), and filler type (X ₃) on the percentage drug released at 8, 12, and 24 h (Y ₁, Y ₂, and Y ₃) as dependent variables. Sixteen TBS sustained-release matrices (F1–F16) were prepared by melt solid dispersion method. None of the prepared matrices achieved the targeted release profile. However, F2 that showed a relatively promising drug release was subjected to trial and error optimization for the filler composition to develop two optimized matrices (F17 and F18). F18 which consisted of drug–Compritol®888 ATO at ratio (1:6 w/w) and Avicel PH 101/dibasic calcium phosphate mixture of 2:1 (w/w) was selected as sustained-release layer. TBS bi-layer tablets were evaluated for their physical properties, in vitro drug release, effect of storage on drug content, and in vivo performance in rabbits. The bi-layer tablets showed acceptable physical properties and release characteristics. In vivo absorption in rabbits revealed initial high TBS plasma levels followed by sustained levels over 24 h compared to immediate-release tablets.     

ALGORITHM FOR APPLICATION OF FOURIER ANALYSIS FOR BIORHYTHMIC BASELINES OF PHARMACODYNAMIC INDIRECT RESPONSE MODELS

The change of an indirect pharmacological response R(t) can be described by a periodic time-dependent production rate k_in (t) and a first-order loss constant k_out. If k_in(t) follows some biological rhythm (e.g., circadian), then the response R(t) also displays a periodic behavior. A new approach for describing the input function in indirect response models with biorhythmic baselines of physiologic substances is introduced. The present approach uses the baseline (placebo) response R_b(t) to recover the equation for k_in(t). Fourier analysis provides an approximate equation for R_b(t) that consists of terms (usually two or three) of the Fourier series (harmonics) that contribute most to the overall sum. The model differential equation is solved backward for k_in(t), yielding the equation involving R_b(t). A computer program was developed to perform the square L²-norm approximation technique. Fourier analysis was also performed based on nonlinear regression. Cortisol suppression after inhalation of fluticasone propionate (FP) was modeled based on the inhibition of the secretion rate k_in(t) using ADAPT II. The pharmacodynamic parameters k_out and IC₅₀ were estimated from the model equation with k_in(t) derived by the new approach. The proposed method of describing the input function needs no assumption about the behavior of k_in(t), is as efficient as methods used previously, and is more flexible in describing the baseline data than the nonlinear regression method. (Chronobiology International, 17(1), 77–93, 2000)     

Kinetic Extractions of Nickel and Lead from Some Contaminated Calcareous Soils

The mobility of heavy metals in contaminated soils is dependent on the kinetics release from soils. Metal extraction over time is commonly used to distinguish two or more fractions of metal based on differences of extraction or release rates. Kinetic studies using 0.01 M CaCl₂, 0.01 M malic acid, and 0.01 M EDTA extractions were performed to characterize nickel (Ni) and lead (Pb) kinetic release in 10 contaminated calcareous soils. Proportions of Ni and Pb extracted with EDTA were higher than when using malic acid and CaCl₂, respectively. The release of Ni and Pb was characterized by an initial fast rate followed by a slower rate and could best be described by a two first-order reactions model with rate constants k₁ and k₂ and two metal pools: readily labile (Q ₁) and less labile (Q ₂). In an EDTA extractant, different Q₁ /Q₂ ratios for Ni and Pb were observed, indicating binding energies to soil constituents is not comparable. The k₁ of the model for Ni (average of 10 soils: 0.2204 h^?1 and 0.2359 h^?1 for 0.01 M CaCl₂ and 0.01 EDTA, respectively) was higher than Pb (0.1044 h^?1 and 0.1631 h^?1 for 0.01 M CaCl₂ and 0.01 M EDTA, respectively), indicating a higher potential of Ni for leaching and groundwater contamination in contaminated calcareous soils. Relationships between the fraction of Ni and Pb determined with the two first-order reactions model and the soil composition and Pb fractions were established. The results indicated that the efficiency of the extractions Ni and Pb from soils depends both on the Ni and Pb content and soil composition. Overall, the results indicated that the use of a 0.01 M malic acid washing solution would be preferred in the field condition.     

Efficient biokinetic experimental designs

Of the experimental methods available for obtaining data to estimate the biological kinetic parameters μ_m, K_s, and Yeach requires considerable experimental effort, yet often yields somewhat imprecise estimates of the parameters, particularly K_s. Therefore it would be worthwhile to seek ways to get parameter estimates of greater precision using less experimental effort. The precision of parameter estimates is strongly dependent, upon the settings of the independent, variables used in the experiments. This dependence is explained and an attempt made to show how experimental settings can be determined that lead efficiently to precise parameter estimates with minimal effort.     

Estimation of Precision in DLW Studies Using the Two-Point Methodology

Previous attempts to estimate precision of doubly labeled water (DLW) estimates of CO₂ production, using propagation of error analyses, have necessarily made simplifying assumptions which may compromise the resultant error estimate. Using an empirical iteration approach, error distributions for the DLW calculation were generated which overcome these problems. The error distribution for CO₂ estimates generated by DLW is symmetrical but not normal. The distribution is significantly truncated such that the 99% inclusion limits are 2.034 SD and not 2.58 SD. The precision error (99% CI for mean as percent of the mean) in DLW experiments, when using duplicate analyses, varies between approximately 3% and 47% depending on the ratio of the elimination constants of the two labels (k_o/k_d), experimental duration and initial isotope dose. The error could be improved by approximately 10 fold by increasing the number of replicates at all six isotope determinations from 2 to 5. Estimating precision in actual experiments can be made using the same empirical approach. The resultant estimates can be of extreme value in evaluating the role of precision as a factor influencing deviations during validation studies, and also for weighting mean estimates in applications of the technique. The deviations of DLW estimates from those made simultaneously by indirect calorimetry in a small mammal validation study were generally greater than the precision of the DLW estimates of CO₂ production. This may indicate there are more problems with the technique than precision alone.     

Evaluation of the Impacts of Formulation Variables and Excipients on the Drug Release Dynamics of a Polyamide 6,10-Based Monolithic Matrix Using Mathematical Tools

Drug release from hydrophilic matrices is regulated mainly by polymeric erosion, disentanglement, dissolution, swelling front movement, drug dissolution and diffusion through the polymeric matrix. These processes depend upon the interaction between the dissolution media, polymeric matrix and drug molecules, which can be significantly influenced by formulation variables and excipients. This study utilized mathematical parameters to evaluate the impacts of selected formulation variables and various excipients on the release performance of hydrophilic polyamide 6,10 (PA 6,10) monolithic matrix. Amitriptyline HCl and theophylline were employed as the high and low solubility model drugs, respectively. The incorporation of different excipient concentrations and changes in formulation components influenced the drug release dynamics as evidenced by computed mathematical quantities (t _x%, MDT _x%, f ₁, f ₂, k ₁, k ₂, and К _F). The effects of excipients on drug release from the PA 6,10 monolithic matrix was further elucidated using static lattice atomistic simulations wherein the component energy refinements corroborates the in vitro and in silico experimental data. Consequently, the feasibility of modulating release kinetics of drug molecules from the novel PA 6,10 monolithic matrix was well suggested.     

A new estimation of global soil greenhouse gas fluxes using a simple data-oriented model

Soil greenhouse gas fluxes (particularly CO₂, CH₄, and N₂O) play important roles in climate change. However, despite the importance of these soil greenhouse gases, the number of reports on global soil greenhouse gas fluxes is limited. Here, new estimates are presented for global soil CO₂ emission (total soil respiration), CH₄ uptake, and N₂O emission fluxes, using a simple data-oriented model. The estimated global fluxes for CO₂ emission, CH₄ uptake, and N₂O emission were 78 Pg C yr⁻¹ (Monte Carlo 95% confidence interval, 64–95 Pg C yr⁻¹), 18 Tg C yr⁻¹ (11–23 Tg C yr⁻¹), and 4.4 Tg N yr⁻¹ (1.4–11.1 Tg N yr⁻¹), respectively. Tropical regions were the largest contributor of all of the gases, particularly the CO₂ and N₂O fluxes. The soil CO₂ and N₂O fluxes had more pronounced seasonal patterns than the soil CH₄ flux. The collected estimates, including both the previous and the present estimates, demonstrate that the means of the best estimates from each study were 79 Pg C yr⁻¹ (291 Pg CO₂ yr⁻¹; coefficient of variation, CV = 13%, N = 6) for CO₂, 21 Tg C yr⁻¹ (29 Tg CH₄ yr⁻¹; CV = 24%, N = 24) for CH₄, and 7.8 Tg N yr⁻¹ (12.2 Tg N₂O yr⁻¹; CV = 38%, N = 11) for N₂O. For N₂O, the mean of the estimates that was calculated by excluding the earliest two estimates was 6.6 Tg N yr⁻¹ (10.4 Tg N₂O yr⁻¹; CV = 22%, N = 9). The reported estimates vary and have large degrees of uncertainty but their overall magnitudes are in general agreement. To further minimize the uncertainty of soil greenhouse gas flux estimates, it is necessary to build global databases and identify key processes in describing global soil greenhouse gas fluxes.     

Physiological control and process kinetics of clavine alkaloid production byClaviceps purpurea

Summary Kinetic parameters of production of clavine alkaloids were evaluated in twoClaviceps purpurea strains. Mutagenesis brought about enhanced resistance of the biosynthetic system towards alkaloids. Addition of glucose into the fermentation medium altered the zero order kinetics of production to activation-inhibition kinetics. The glucose treatment allowed performance of both elymoclavine-inhibitionless and clavine alkaloid-decompositionless fermentations if a combination of fermentation and separation units in a closed loop was used.Nomenlacture k ₁ rate constant of agroclavine synthesis (mg Agro · mg Elymo/l·g DW·day for stage 1, mg Agro/g DW·day for stage 2) - k ₂ parameter describing inhibition of agroclavine formation rate by elymoclavine (mg Elymo/l) - k ₃ specific rate of agroclavine decay (l/g DW·day) - k ₄ maximal specific rate of elymoclavine synthesis (stage 1, 1/g DW·day, stage 2, mg Elymo/g DW·day) - k ₄ ^– maximal specific rate of elymoclavine synthesis in stage 1 (inhibition-activation mechanism) (mg Elymo/g DW·day) - k ₅ physiological constant describing the elymoclavine decay rate (l²/g DW·day·mg Elymo) - k ₅ ^– physiological constant describing the activation of elymoclavine biosynthesis by elymoclavine (mg Elymo/l) - k ₆ physiological constant describing the repression of elymoclavine biosynthesis by elymoclavine (mg Elymo/l) - k ₇ maximal specific growth rate (1/day) - k ₈ specific rate of biomass decay (l/g DW·day) - A agroclavine concentration (mg/l) - E elymoclavine concentration (mg/l) - r _A specific rate of agroclavine biosynthesis (mg Agro/g DW·day) - r _E specific rate of elymoclavine biosynthesis (mg Elymo/g DW·day) - r _i specific rate of alkaloid biosynthesis (mg alkaloid/g DW·day) - X dry biomass concentration (g/l) - specific growth rate (1/day) Abbreviations Agro agroclavine - Elymo elymoclavine - Chano chanoclavine - DW dry weight of biomass     

Evaluation of a vermicompost and leachates on <i>Solidago x hybrida</i> and organic carbon mineralization under aerobic incubations

In the floriculture region of Tenancingo in the State of Mexico, the application of stabilized organic matter, such as vermicompost and leachates, contributes to improve the quality of the soil and plant nutrition. However, it is important to know the chemical composition of a vermicompost and the mineralization process. This is because the amount and speed of nutrient release which will be available to the crop will depend on that knowledge. The objective of the study was to evaluate the effect of the application of a vermicompost and leachates on various quantitative variables of Solidago x hybrid, and the mineralization of organic carbon under aerobic incubations. Vermicompost (74 and 36 g/kg soil), leachates (5 and 10 L/kg soil), 0.33 g/kg soil of chemical fertilizer Ca (NO₃)₂, and not treated soil (control) were applied under greenhouse conditions to evaluate their effects on plant growth variables. Mixtures of 100 g of soil with vermicompost and leachates were made in the laboratory which were incubated during 9 weeks to obtain the potentially mineralizable organic carbon (Corg PM) and the rate of mineralization (k) after adjusting an exponential model. In the greenhouse experiment there were no statistical differences after applying vermicompost and leachates on the quantitative variables (number of stems per plant, diameter of the panicle, fresh weight, plant length and stem diameter) with respect to the control (p>0.05). The effect among the applied doses was evident only for variables such as fresh weight, panicle length and stem diameter with respect to the control. In incubated soils, k values ranged between 0.209-0.325 C mg/kg soil/week. Only with the application of leachates in high doses two pools of organic matter were shown: one soluble labile (102.9 mg/kg soil) and the other hydrolysable (819 mg/kg soil). The soluble, labile fraction favored nutrient availability immediately after its application to the soil. However, a single pool of hydrolysable organic C (987-1074 mg/kg soil) was found when vermicompost was applied. It was associated with a release of organic matter during crop development, and to a possible stimulation of microbial activity. High values of electrical conductivity in vermicompost and leachates (8.2-11.7 mS/m) suggest a moderate application of both products.     

The rate of permafrost carbon release under aerobic and anaerobic conditions and its potential effects on climate

Recent observations suggest that permafrost thaw may create two completely different soil environments: aerobic in relatively well‐drained uplands and anaerobic in poorly drained wetlands. The soil oxygen availability will dictate the rate of permafrost carbon release as carbon dioxide (CO₂) and as methane (CH₄), and the overall effects of these emitted greenhouse gases on climate. The objective of this study was to quantify CO₂ and CH₄ release over a 500‐day period from permafrost soil under aerobic and anaerobic conditions in the laboratory and to compare the potential effects of these emissions on future climate by estimating their relative climate forcing. We used permafrost soils collected from Alaska and Siberia with varying organic matter characteristics and simultaneously incubated them under aerobic and anaerobic conditions to determine rates of CO₂ and CH₄ production. Over 500 days of soil incubation at 15 °C, we observed that carbon released under aerobic conditions was 3.9–10.0 times greater than anaerobic conditions. When scaled by greenhouse warming potential to account for differences between CO₂ and CH₄, relative climate forcing ranged between 1.5 and 7.1. Carbon release in organic soils was nearly 20 times greater than mineral soils on a per gram soil basis, but when compared on a per gram carbon basis, deep permafrost mineral soils showed carbon release rates similar to organic soils for some soil types. This suggests that permafrost carbon may be very labile, but that there are significant differences across soil types depending on the processes that controlled initial permafrost carbon accumulation within a particular landscape. Overall, our study showed that, independent of soil type, permafrost carbon in a relatively aerobic upland ecosystems may have a greater effect on climate when compared with a similar amount of permafrost carbon thawing in an anaerobic environment, despite the release of CH₄ that occurs in anaerobic conditions.     

Persistence of Viruses in Desert Soils Amended with Anaerobically Digested Sewage Sludge

Pima County, Ariz., is currently investigating the potential benefits of land application of sewage sludge. To assess risks associated with the presence of pathogenic enteric viruses present in the sludge, laboratory studies were conducted to measure the inactivation rate (k = log₁₀ reduction per day) of poliovirus type 1 and bacteriophages MS2 and PRD-1 in two sludge-amended desert agricultural soils (Brazito Sandy Loam and Pima Clay Loam). Under constant moisture (approximately -0.05 × 10⁵ Pa for both soils) and temperatures of 15, 27, and 40°C, the main factors controlling the inactivation of these viruses were soil temperature and texture. As the temperature increased from 15 to 40°C, the inactivation rate increased significantly for poliovirus and MS2, whereas, for PRD-1, a significant increase in the inactivation rate was observed only at 40°C. Clay loam soils afforded more protection to all three viruses than sandy soils. At 15°C, the inactivation rate for MS2 ranged from 0.366 to 0.394 log₁₀ reduction per day in clay loam and sandy loam soils, respectively. At 27°C, this rate increased to 0.629 log₁₀ reduction per day in clay loam soil and to 0.652 in sandy loam soil. A similar trend was observed for poliovirus at 15°C (k = 0.064 log₁₀ reduction per day, clay loam; k = 0.095 log₁₀ reduction per day, sandy loam) and 27°C (k = 0.133 log₁₀ reduction per day, clay loam; k = 0.154 log₁₀ reduction per day, sandy loam). Neither MS2 nor poliovirus was recovered after 24 h at 40°C. No reduction of PRD-1 was observed after 28 days at 15°C and after 16 days at 27°C. At 40°C, the inactivation rates were 0.208 log₁₀ reduction per day in amended clay loam soil and 0.282 log₁₀ reduction per day in sandy loam soil. Evaporation to less than 5% soil moisture completely inactivated all three viruses within 7 days at 15°C, within 3 days at 27°C, and within 2 days at 40°C regardless of soil type. This suggests that a combination of high soil temperature and rapid loss of soil moisture will significantly reduce risks caused by viruses in sludge.     

The isomerization of D-glucose into D-fructose catalyzed by whole-cell immobilized glucose isomerase. The dependence of the intrinsic rate of reaction on substrate concentration,pH, and temperature

The relation between the intrinsic rate of the glucose-fructose isomerization catalyzed by whole-cell immobilized glucose isomerase and the substrate concentration can be described with the kinetic model The numerical values of k₁, k_?1, k₂, and k_?2 have been determined from low-conversion experiments starting from pure glucose or fructose solutions, and are presented as a function of pH and temperature. The difference between the overall chemical reaction rate determined in high-conversion experiments and that calculated from the individual k₁, k_?1, k₂, and k_?2 values is less than 10%.     

Variation of ADM1 by using temperature-phased anaerobic digestion (TPAD) operation

The objective of the study was to examine the application of the Anaerobic Digestion Model No. 1 (ADM1) developed by the IWA task group for mathematical modelling of anaerobic process. Lab-scale temperature-phased anaerobic digestion (TPAD) process were operated continuously, and were fed with co-substrate composed of dog food and flour. The model platform implemented in the simulation was a derivative of the ADM1. Sensitivity analysis showed that k_m.process (maximum specific uptake rate) and K_S.process (half saturation value) had high sensitivities to model components. Important parameters including maximum uptake rate for propionate utilisers (k_m.pro) and half saturation constant for acetate utilisers (K_S.ac) in the thermophilic digester and maximum uptake rate for acetate utilisers (k_m.ac) in the mesophilic digester were estimated using iterative methods, which optimized the parameters with experimental results. Simulation with estimated parameters showed good agreement with experimental results in the case of methane production, uptake of acetate, soluble chemical oxygen demand (SCOD) and total chemical oxygen demand (TCOD). Under these conditions, the model predicted reasonably well the dynamic behavior of the TPAD process for verifying the model.