Optimization of the Emerson–Trinder enzymatic reaction by response surface methodology

The Emerson–Trinder reaction has been optimized in this work using an initial rate spectrophotometric method and response surface methodology (RSM). In this investigation, the variation range of critical variables along with the fixed parameters were selected based on a preliminary ‘one at a time’ (OVAT) procedure for the subsequent RSM chemometric analysis as follows: pH (6–10), buffer concentration (50–250?mM), 4-aminoantipyrine (4-AAP) concentration (1–5?mM), temperature (25–45°C). The optimum values of fixed parameters were: 4-fluorophenol (4-FP, 30?mM), horseradish peroxidase (HRP) enzyme activity (0.12?U?mL^?1), and the fixed concentration of the H₂O₂ in the chemometric experiments was 11.4 µM. The non-linear nature of the experimental response of the reaction system was explained by a second-order polynomial equation, which revealed the impact of the experimental factors, their interactions and also their optimum values. The results of the reported RSM analysis proved to be quite appropriate for the design and optimization of this reaction, as illustrated by the relatively high value of the determination coefficient (R²=96.7%) for the fitting of quadratic model, along with the satisfactory results generated by the analysis of variance (ANOVA). All the evaluated analytical characteristics of this method: typical reaction progress curves, resulting linear calibration curve, within-day precisions at low and at high levels, and the upper and lower detection limits were, also, reported. In addition, to check the quality of the optimization and validity of the model, the assay of H₂O₂, in pooled serum matrix and in cosmetic samples, was performed.     

Modeling in vitro cholesterol reduction in relation to growth of probiotic Lactobacillus casei

Lactobacillus casei LA‐1 isolated from a nondairy fermented source was evaluated for its in vitro ability to reduce cholesterol. The bacterium tested positive for bile salt deconjugation in relation to cholesterol removal. Tested growth‐associated physiological variables such as pH, temperature and inoculum size were all found to have significant effects on in vitro cholesterol reduction and biomass production (both P < 0.005). Furthermore, a central composite design was used to evaluate the effects of significant variables and their interactions. A linear regression model was developed for in vitro cholesterol reduction as a function of growth‐associated variables. Maximum cholesterol reduction achieved was 45% whereas maximum biomass yield of 2.34 optical density was observed at the central point. Our study possibly indicates that the growth of L. casei LA‐1 depends on its cholesterol removing ability.     

CALL ANALYSIS OF TRIPLOID HYBRIDS RESULTING FROM DIPLOID-TETRAPLOID SPECIES CROSSES OF HYLID TREE FROGS

ABSTRACT

Calls produced by hybrids resulting from laboratory crosses of tetraploid Hyla versicolor females and either diploid Hyla chrysoscelis (type I) or Hyla arborea (type II) males were induced through manipulation of environmental conditions. Type I hybrids produced trilled calls similar in note repetition to H. versicolor, but more similar in dominant frequency to H. chrysoscelis. Mean duration was shorter than in both parent calls. Type II hybrids produced calls which were longer in duration and lower in note repetition rate than H. versicolor, but shorter in duration and higher in note repetition rate than H. arborea. Dominant frequency of type II hybrids was lower than in H. arborea but not significantly different than in H. versicolor. Hybrid calls were not strictly intermediate, and may provide information regarding parental relationships.     

Isolation and characterization of Aspergillus sp. for the production of extracellular polysaccharides by response surface methodology

In this study, Aspergillus sp. was isolated for the production of extracellular polysaccharide. The process parameters were initially optimized by traditional methods. The cheap substrate, wheat bran was used for the production of extracellular polysaccharide in solid state fermentation. Supplementation of (1%, w/w) maltose, gelatin enhanced EPS production (5.36?mg/g). The salts such as, Cu²⁺ (4.9?mg/g), Ca²⁺ (3.5?mg/g), Zn²⁺ (2.9?mg/g), Mn²⁺ (3.4?mg/g) and Mg²⁺ (1.8?mg/g) stimulated EPS production. In two level full factorial experimental designs, the EPS yield varied from 3.18 to 11.65?mg/g wheat bran substrate with various combinations of the components supplemented with wheat bran substrate. Among these selected factors in central composite design, maltose significantly influenced on extracellular polysaccharide production.     

Response surface method was used to optimize the ultrasonic assisted extraction of flavonoids from Crinum asiaticum

In this study, ultrasound-assisted extraction of flavonoid from Crinum asiaticum was studied through response surface methodology (RSM) to gain the best extraction process of flavonoid and enhance the extraction rate of flavonoid. In the following RSM experiment, we selected the corresponding data of every factor as the center point through the single-factor experiments, then the experimental data was subjected to multiple regression analysis. According to the statistical analysis results, the results were consistent with the polynomial regression model, the determination coefficient (R2) was 0.9769. The best conditions for maximum flavonoid yield were 60% ethanol concentration, 64 °C for extraction temperature, 1:28 (v/w) solid-to-liquid ratio with extraction time for 47 min. The best response of flavonoid yield was 1.63972%. The predicted results for best reaction conditions were in good agreement with experiment values. Ultrasound-assisted extraction method can enhance the extraction rate of flavonoid significantly. It is a powerful tool to extract of important phytochemicals from nature plant.     

Aggregation of measures to produce an overall assessment of animal welfare. Part 1: a review of existing methods

Several systems have been proposed for the overall assessment of animal welfare at the farm level for the purpose of advising farmers or assisting public decision-making. They are generally based on several measures compounded into a single evaluation, using different rules to assemble the information. Here we discuss the different methods used to aggregate welfare measures and their applicability to certification schemes involving welfare. Data obtained on a farm can be (i) analysed by an expert who draws an overall conclusion; (ii) compared with minimal requirements set for each measure; (iii) converted into ranks, which are then summed; or (iv) converted into values or scores compounded in a weighted sum (e.g. TGI35L) or using ad hoc rules. Existing methods used at present (at least when used exclusively) may be insufficiently sensitive or not routinely applicable, or may not reflect the multidimensional nature of welfare and the relative importance of various welfare measures. It is concluded that different methods may be used at different stages of the construction of an overall assessment of animal welfare, depending on the constraints imposed on the aggregation process.     

Synthesis of novel chalcones through palladium-catalyzed CO cross-coupling reaction of bromo-chalcones with ethyl acetohydroxamate and their antiplasmodial evaluation against Plasmodium falcipuram in vitro

An efficient method for palladium-catalyzed CO cross-coupling of ethyl acetohydroxamate (EAcHO) with 4-bromo-chalcones has been developed to synthesize novel chalcones. The two supporting ligands, namely tBuXPhos (L7), and cataCXium®PIntB (L16) were found to be effective ligands towards the Pd-catalyzed CO cross-coupling reaction to afford the desired product in moderate to excellent yields (50–99%). The coupled products were screened for in vitro blood stage antiplasmodial activity against Plasmodium falciparum (3D7) using the [³H] hypoxanthine incorporation inhibition assay. Of the twenty two compounds screened, eleven showed good antiplasmodial activity with IC₅₀ values ranging from 6–16 μg/mL. The selected active molecules 11, 16, 22, (IC₅₀ 12 μg/mL) and 19 (IC₅₀ 6 μg/mL) were studied for their cytotoxic effect against HepG2 Cells (human hepatocellular liver carcinoma cell lines), showing the selectivity index (SI) values are greater than 4 except chalcone 22. Our result demonstrates a methodology for synthesizing novel chalcones as a new class of antiplasmodial agent.     

Does the Spanish version of the SWLS measure the same in Spain and Peru?

AimSatisfaction with life is a measure of protection in older adults. There lies the importance of providing quality instruments. The aim of the study was to evaluate the invariance of the life satisfaction scale (SWLS) in two samples of older adults in Spain and Peru.MethodThe participants were 857 older adults in Spain (mean age = 68.23 years, SD = 5.93) and 336 older adults in Peru (average age = 72.42, SD = 7.07). All multi-group confirmatory factor analyzes were estimated in Mplus 8.0.ResultsThe results indicate the presence of a strict invariance of the one-dimensional structure of the SWLS in samples of older adults in Spain and Peru, which allows for meaningful comparisons of latent means and covariances. Comparison of latent means showed small differences in the construct between the cultural groups.ConclusionsThe SWLS is a valid instrument for intercultural comparisons between Spanish and Peruvian population. The measurement invariance assessment contributes to a better understanding of life satisfaction in populations from different cultural contexts.     

Estimates of the residual nitrogen benefit of groundnut to maize in Northeast Thailand

Four cultivars of groundnut were grown in upland soil in Northeast Thailand to study the residual benefit of the stover to a subsequent maize crop. An N-balance estimate of the total residual N in the maize supplied by the groundnut was made. In addition three independent estimates were made of the residual benefits to maize when the groundnut stover was returned to the land and incorporated. The first estimate (Estimate 1) was an N-balance estimate. A dual labelling approach was used where ¹⁵N-labelled stover was added to unlabelled microplots (Estimate 2) or unlabelled stover was added to ¹⁵N-labelled soil microplots (Estimate 3). The nodulating groundnut cultivars fixed between 59–64% of their nitrogen (as estimated by the ¹⁵N isotope dilution method using non-nodulating groundnut as a non-fixing reference) producing between 100 and 130 kg N ha^-1 in their stover. Although the following maize crop suffered from drought stress, maize grain N and dry weights were up to 80% and 65% greater respectively in the plots where the stover was returned as compared with the plots where the stover was removed. These benefits were comparable with applications of 75 kg N ha^-1 nitrogen in the form of urea. The total residual N estimates of the contribution of the nodulated groundnut to the maize ranged from 16.4–27.5 kg N ha^-1. Estimates of the residual N supplied by the stover and fallen leaves ranged from 11.9–21.3 kg N ha^-1 using the N-balance method (Estimate 1), from 6.3–9.6 kg N ha^-1 with the labelled stover method (Estimate 2) and from 0–11.4 kg N ha^-1 with the labelled soil method. There was closest agreement between the two ¹⁵N based estimates suggesting that apparent added nitrogen interactions in these soils may not be important and that N balance estimates can overestimate the residual N in crops following legumes, even in very poor soils. This work also indicates the considerable ability of local groundnut cultivars to fix atmospheric nitrogen and the potential benefits from returning and incorporating legume residues to the soil in the upland cropping systems of Northeast Thailand. The applicability of the ¹⁵N methodology used here and possible reasons for the discrepancies between estimates 1, 2 and 3 are discussed.