Constrained dansyl derivatives reveal bacterial specificity of highly conserved thymidylate synthases

The elucidation of the structural/functional specificities of highly conserved enzymes remains a challenging area of investigation, and enzymes involved in cellular replication are important targets for functional studies and drug discovery. Thymidylate synthase (TS, ThyA) governs the synthesis of thymidylate for use in DNA synthesis. The present study focused on Lactobacillus casei TS (LcTS) and Escherichia coli TS (EcTS), which exhibit 50 % sequence identity and strong folding similarity. We have successfully designed and validated a chemical model in which linear, but not constrained, dansyl derivatives specifically complement the LcTS active site. Conversely, chemically constrained dansyl derivatives showed up to 1000-fold improved affinity for EcTS relative to the inhibitory activity of linear derivatives. This study demonstrates that the accurate design of small ligands can uncover functional features of highly conserved enzymes.     

Discontinuous carbon fiber/polyamide composites with microencapsulated paraffin for thermal energy storage

This work focuses on the development of multifunctional thermoplastic composites with thermal energy storage capability. A polyamide 12 (PA12) matrix was filled with a phase change material (PCM), constituted by paraffin microcapsules (T_melt = 43 °C), and reinforced with carbon fibers (CFs) of two different lengths (chopped/CF “long”[CFL] and milled/CF “short” [CFS]). DSC tests showed that the melting/crystallization enthalpy values increase with the PCM weight fraction up to 60 J/g. The enthalpy was 41–94% of the expected value and decreased with an increase in the fiber content, because the capsules were damaged by the increasing viscosity and shear stresses during compounding. Long CFs increased the elastic modulus (+316%), tensile strength (+26%), and thermal conductivity (+54%) with respect to neat PA12. Thermal imaging tests evidenced a slower cooling for the samples containing PCM, and once again the CFS-containing samples outperformed those with CFL, due to the higher effective PCM content. © 2018 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2019 , 136, 47408.     

Optimization and Single-Stage Centrifugal Contactor Experiments with the Novel Hydrophilic Complexant PyTri-Diol for the i-SANEX Process

The applicability of 2,6-bis[1-(propan-1-ol)-1,2,3-triazol-4-yl)]pyridine (PyTri-Diol) in the innovative Selective ActiNide EXtraction (i-SANEX) process as water-soluble complexing agent was studied. Preliminary batch experiments were aimed at identifying the optimal formulation of the PyTri-Diol solution and at preparing the ground for single-stage centrifugal contactor experiments. A N,N,N′,N′ tetraoctyl diglycolamide (TODGA)-based solvent loaded using a spiked synthetic Plutonium and Uranium Reduction EXtraction raffinate and the optimized PyTri-Diol aqueous phase were contacted in a single-stage annular centrifugal contactor setup with three different flow-rate conditions. No hydrodynamic problems were encountered, and promising minor actinide separation from other cations was achieved with satisfactory kinetics and stage efficiency. The flow sheet of a TODGA–PyTri-Diol-based i-SANEX process was designed exploiting batch and single-stage data, promoting the CHON compliant PyTri-Diol as excellent alternative to the formerly used SO₃–Ph–BTP.     

Optical trapping of porous silicon nanoparticles

Silicon nanoparticles obtained by ball-milling of a 50% porosity silicon layer have been optically trapped when dispersed in a water-surfactant environment. We measured the optical force constants using linearly and radially polarized trapping beams finding a reshaping of the optical potential and an enhanced axial spring constant for the latter. These measurements open perspectives for the control and handling of silicon nanoparticles as labeling agents in biological analysis and fluorescence imaging techniques.     

PEG-templated mesoporous silica nanoparticles exclusively target cancer cells

Mesoporous silica nanoparticles (MSNs) have been proposed as DNA and drug delivery carriers, as well as efficient tools for fluorescent cell tracking. The major limitation is that MSNs enter cells regardless of a target-specific functionalization. Here we show that non functionalized MSNs, synthesized using a PEG surfactant-based interfacial synthesis procedure, do not enter cells, while a highly specific, receptor mediated, cellular internalization of folic acid (FOL) grafted MSNs (MSN-FOL), occurs exclusively in folate receptor (FR) expressing cells. Neither the classical clathrin pathway nor macropinocytosis is involved in the MSN endocytic process, while fluorescent MSNs (MSN-FITC) enter cells through aspecific, caveolae-mediated, endocytosis. Moreover, internalized particles seem to be mostly exocytosed from cells within 96 h. Finally, cisplatin (Cp) loaded MSN-FOL were tested on cancerous FR-positive (HeLa) or normal FR-negative (HEK293) cells. A strong growth arrest was observed only in HeLa cells treated with MSN-FOL-Cp. The results presented here show that our mesoporous nanoparticles do not enter cells unless opportunely functionalized, suggesting that they could represent a promising vehicle for drug targeting applications.     

In vitro fermentability of differently digested resistant starch preparations

The in vitro fermentability of two resistant starch preparations type 2 (RS2) and type 3 (RS3) was investigated using human colonic microbiota. Prior to the fermentation experiments, samples were digested using two in vitro models, a batch (ba) and a dynamic (dy), as well as an in vivo method (il) for RS3. Digestion residues were fermented in vitro using a simple batch model lasting 24 h and a more sophisticated dynamic model enduring 72 h. During batch fermentation, metabolite productions and starch degradation rates were similar for RS2 and RS3 but higher for dy- compared to ba-digested samples. RS3il led to the lowest fermentability. Furthermore, increased butyrate ratios were observed for all preparations. The varying RS preparations behaved similarly in the dynamic fermentation but showed high SDs. Moreover, the fermentability was slow during the first 24 h, indicating that the microbiota needed an adaptation period to ferment RS. Propionate ratios increased at the expense of butyrate with exception for RS2dy showing an increase in acetate only. Differences in fermentability observed between the dynamic model, allowing a closer simulation of the in vivo behavior, and the batch model, recommended for screening purposes, could be due to the varying microbiota used.     

Fate of surfactants in membrane bioreactors and conventional activated sludge plants

Two membrane bioreactors (MBRs) were operated at high sludge retention time (SRT) (between 30 and 75 d) in parallel to a conventional activated sludge plant (CASP) conducted at SRT = 10 d. The fate of linear alkylbenzene sulfonate (LAS), nonylphenol ethoxylates (NP(n)EO, n = 1-15), nonylphenoxy carboxylates (NP(n)EC, n = 1-2), and nonylphenol (NP) in these systems was investigated. All systems were very efficient in the removal of LAS (around 99%). The analysis of variance showed that the difference in the removal efficiency of LAS in the CASP and the MBR operated at SRT = 65-75 d (respectively 99.0 ± 0.43 and 99.8 ± 0.11) were significant (p < 0.05), confirming the importance of SRT in the removal of LAS. Comparison between the CASP and the MBRs in the removal efficiency of nonylphenolic compounds were conducted considering NP(3-15)EO, the sum of NP(1-15)EO, NP(1-2)EC, and nonylphenol (NP). In all cases MBRs were more efficient than the CASP. In the case of NP the removal was about 76 ± 7.5% for the CASP and 90% ± 12.1 and 82 ± 8.7% for the MBRs. Better performance of MBRs in the removal of nonylphenolic compounds can be attributed to a better degradation. For example, if the sum of NP(1-15)EO and NP(1-2)EC is considered, estimated biodegradation was about 48% for the CASP and 72% for MBRs.     

Lactation curves of Sarda breed goats estimated with test day models

Test day records of milk yield (38,765), fat and protein contents (11,357) of Sarda goats (the most numerous Italian goat breed) were analysed with mixed linear models in order to estimate the effects of test date (month and year of kidding for fat and protein contents) parity, number of kids born, altitude of location of flocks (<200 m asl, 200-500 m asl, >500 m asl), flocks within altitude and lactation stage (eight days-in-milk intervals of 30 d each) on milk production. All factors considered in the models affected milk traits significantly. Milk yield was lower in first parity goats than in higher parities whereas fat and protein contents showed an opposite trend. Goats with two kids at parturition had a higher milk yield than goats with one kid and tended to have lower fat and protein percentages. Repeatability between test days within lactation was 0.34, 0.17 and 0.45 for milk yield, fat content and protein content, respectively. Lactation curves of goats farmed at different altitudes were clearly separated, especially for milk yield. Results of the present study highlight differences in milk production traits among the three subpopulations that have been previously identified within the Sarda breed on the basis of the morphological structure of animals and altitude of location of flocks.     

Abundance of mRNA of apolipoprotein b100, apolipoprotein e, and microsomal triglyceride transfer protein in liver from periparturient dairy cows

Limited secretion of very low density lipoproteins (VLDL) in dairy cows is strongly related to fatty liver and other metabolic disorders in the early postpartum. Currently, there is limited information on which roles apolipoprotein B(100) (ApoB(100)), apolipoprotein E (ApoE), and microsomal triglyceride transfer protein (MTP) play in that VLDL limitation. To our knowledge, no studies have simultaneously measured ApoB(100), ApoE, and MTP mRNA in periparturient dairy cows. Therefore, a trial was conducted to assess liver gene expression of these proteins in transition dairy cows and to evaluate the relationships between their expression and metabolic status. Eight multiparous Holstein cows were monitored during the transition period. To evaluate metabolic and nutritional status, body condition score was registered, and plasma indexes of energy metabolism and VLDL were determined from 35 d before to 35 d after calving. Liver biopsies were performed on d -35, 3, and 35 relative to day of calving, and gene expression of ApoB(100), ApoE, and MTP were determined on liver tissue. Body condition, plasma glucose and VLDL decreased, and plasma NEFA and BHBA increased after calving. Compared with values of d -35, on d 3 after calving the ApoB(100) mRNA synthesis was lower, whereas MTP and ApoE mRNA abundance were higher. Negative correlation (r = -0.57) between plasma NEFA concentration and ApoB(100) mRNA abundance, and positive correlation between ApoB(100) mRNA abundance and plasma cholesterol (r = 0.65) and plasma albumins (r = 0.52) were detected at 3 d postpartum. Data on changes of gene expression of the 3 main proteins involved in the regulation of synthesis and secretion of VLDL in the liver suggest that decreased mRNA for ApoB(100) may be consistent with decreased synthesis and/or secretion of VLDL from liver during the periparturient period.     

Optimal offering and allocation policies for wind power in energy and reserve markets

Proliferation of wind power generation is increasingly making this power source an important asset in designs of energy and reserve markets. Intuitively, wind power producers will require the development of new offering strategies that maximize the expected profit in both energy and reserve markets while fulfilling the market rules and its operational limits. In this paper, we implement and exploit the controllability of the proportional control strategy. This strategy allows the splitting of potentially available wind power generation in energy and reserve markets. In addition, we take advantage of better forecast information from the different day‐ahead and balancing stages, allowing different shares of energy and reserve in both stages. Under these assumptions, different mathematical methods able to deal with the uncertain nature of wind power generation, namely, stochastic programming, with McCormick relaxation and piecewise linear decision rules are adapted and tested aiming to maximize the expected revenue for participating in both energy and reserve markets, while accounting for estimated balancing costs for failing to provide energy and reserve. A set of numerical examples, as well as a case study based on real data, allow the analysis and evaluation of the performance and behavior of such techniques. An important conclusion is that the use of the proposed approaches offers a degree of freedom in terms of minimizing balancing costs for the wind power producer strategically to participate in both energy and reserve markets. Copyright © 2017 John Wiley & Sons, Ltd.