Synthesis and biological activity of a novel inhibitor of dihydroceramide desaturase

A novel mechanism-based dihydroceramide desaturase inhibitor (XM462) in which the substrate C5 methylene group is replaced by a sulfur atom is reported. Dihydroceramide desaturase inhibition occurred both in vitro and in cultured cells with IC(50) values of 8.2 and 0.78 microM, respectively, at a substrate concentration of 10 microM. In vitro experiments showed that XM462 produced a mixed-type inhibition (K(i)=2 microM, alpha=0.83). LC-MS analyses showed that accumulation of endogenous dihydroceramides occurred in cells upon treatment with XM462 in serum-free medium, whereas ceramides built up in controls. In addition, XM462 was found to be metabolised to its 1-glucosyl and 1-phosphocholine derivatives, and to the products of N-deacylation and reacylation with palmitoyl and stearoyl groups. In Jurkat A3 cells cultured in serum-free medium, viability, as the percentage of trypan blue unstained cells in total cells, was reduced upon XM462 treatment (5 microM, 24 h), but not in controls. The interest of this compound is discussed.     

Is brown adipose tissue a store of long chain polyunsaturated fatty acids during postnatal developing brain?

Mammals along their early postnatal period develop a substantial amount of a very active brown adipose tissue (BAT). Through this work we explored the possibility that BAT may function as a long chain polyunsaturated fatty acids reservoir (LC-PUFA) during the rapid growth of brain structures. In new born rats 1, 6, 12 and 20 days old, we analyzed fatty acid percentage of triglycerides (TG) and phospholipid fractions, and the absolute amount of TG. In 6 day old rats we also evaluated the extend of further desaturation of 1-14C linoleic acid administered by intraperitoneal injection. Results demonstrated a drastic increase of TG concentration during experimental period (1,5; 40; 118; 120 mg/g wet weight) and LC-PUFA percentage was higher in "1 and 6" than "12 and 20" days old rats (16-17% vs 5%). Our results showed that BAT stored important amounts of LC-PUFA. On the other hand, 1-14C linoleic acid incorporation was higher in liver than BAT. In contrast, the desaturated products of 1-14C linoleic acid /1-14C linoleic acid ratio was greater in BAT than liver (>4). This could indicate that BAT synthesizes LC-PUFA in addition to store it. In summary we demonstrated than BAT is an important reservoir of LC-PUFA during postnatal brain growth.     

Synthesis of a novel ceramide analogue and its use in a high-throughput fluorogenic assay for ceramidases

Several investigations have shown that acid ceramidase inhibitors are potential antiproliferative and cytostatic drugs for cancer chemotherapy. The combinatorial chemistry approach for the discovery of acid ceramidase inhibitors requires the availability of a high-throughput enzyme assay. The synthesis of a novel fluorogenic ceramidase substrate, and its processing both in vitro and in cultured cells in a microtiter plate layout, are reported in this article. This coumarinic substrate was hydrolyzed in vitro (rat liver lysosomes) with Km and Vmax values of 113 microM and 3.6 pmol min-1 mg-1, respectively. Similarly, hydrolysis occurred in intact cultured cells that overexpressed acidic ceramidase. The assay was validated for the identification and characterization of acidic ceramidase inhibitors by using several alpha-ketoamide ceramide analogues, whose inhibitory activity had been previously described.     

On the existence of solutions to a class of optimization problems

A sufficient condition for the existence and uniqueness of solutions to a class of optimization problems in nonlinear programming form, with strictly convex cost functions, convex inequality and linear equality side constraints, and closed convex constraint sets is studied.     

Optimization of the azo dye Procion Red H-EXL degradation by Fenton's reagent using experimental design

Chemical oxidation by Fenton's reagent of a reactive azo dye (Procion Deep Red H-EXL gran) solution has been optimized making use of the experimental design methodology. The variables considered for the oxidative process optimization were the temperature and the initial concentrations of hydrogen peroxide and ferrous ion, for a dye concentration of 100mg/L at pH 3.5, the latter being fixed after some preliminary runs. Experiments were carried out according to a central composite design approach. The methodology employed allowed to evaluate and identify the effects and interactions of the considered variables with statistical meaning in the process response, i.e., in the total organic carbon (TOC) reduction after 120 min of reaction. A quadratic model with good adherence to the experimental data in the domain analysed was developed, which was used to plot the response surface curves and to perform process optimization. It was concluded that temperature and ferrous ion concentration are the only variables that affect TOC removal, and due to the cross-interactions, the effect of each variable depends on the value of the other one, thus affecting positively or negatively the process response.     

Poly (DL-lactide-co-glycolide) (PLGA) nanoparticles with entrapped trans-cinnamaldehyde and eugenol for antimicrobial delivery applications

Eugenol and trans-cinnamaldehyde are natural compounds known to be highly effective antimicrobials; however, both are hydrophobic molecules, a limitation to their use within the food industry. The goal of this study was to synthesize spherical poly (DL-lactide-co-glycolide) (PLGA) nanoparticles with entrapped eugenol and trans-cinnamaldehyde for future antimicrobial delivery applications. The emulsion evaporation method was used to form the nanoparticles in the presence of poly (vinyl alcohol) (PVA) as a surfactant. The inclusion of antimicrobial compounds into the PLGA nanoparticles was accomplished in the organic phase. Synthesis was followed by ultrafiltration (performed to eliminate the excess of PVA and antimicrobial compound) and freeze-drying. The nanoparticles were characterized by their shape, size, entrapment efficiency, and antimicrobial efficiency. The entrapment efficiency for eugenol and trans-cinnamaldehyde was approximately 98% and 92%, respectively. Controlled release experiments conducted in vitro at 37 °C and 100 rpm for 72 h showed an initial burst followed by a slower rate of release of the antimicrobial entrapped inside the PLGA matrix. All loaded nanoparticles formulations proved to be efficient in inhibiting growth of Salmonella spp. (Gram-negative bacterium) and Listeria spp. (Gram-positive bacterium) with concentrations ranging from 20 to 10 mg/mL. Results suggest that the application of these antimicrobial nanoparticles in food systems may be effective at inhibiting specific pathogens. PRACTICAL APPLICATION: Nanoencapsulation of lipophilic antimicrobial compounds has great potential for improving the effectiveness and efficiency of delivery in food systems. This study consisted of synthesizing PLGA nanoparticles with entrapped eugenol and trans-cinnamaldehyde. By characterizing these new delivery systems, one can understand the controlled-release mechanism and antimicrobial efficiency that provides a foundation that will enable food manufacturers to design smart food systems for future delivery applications, including packaging and processing, capable of ensuring food safety to consumers.     

Lead, Cadmium and Cobalt (Pb, Cd, and Co) Leaching of Glass-Clay Containers by pH Effect of Food

Recent studies have shown that handcrafted glass-clay containers are a health risk because they can be contaminated by heavy metals, which can be transferred to food, thus reaching the human body to potentially cause illness. Therefore, in the present work, we evaluate the leaching of lead, cadmium, and cobalt from glass-clay containers into two types of food: tomato sauce (salsa), and chickpea puree. The containers were obtained from four regions in the Mexican state of Hidalgo. Repetitive extractions from the containers were carried out to quantify the leaching of the heavy metals into the salsa, the chickpea puree, and acetic acid using the technique proposed by the USFDA. The results show that greater use of the containers leads to more leaching of heavy metals into both types of food and into the acetic acid, with the greatest metal extraction recorded for the Ixmiquilpan vessels. These results indicate that the metals present in the glass-clay containers leach into the food and that increased reuse increases the risk to the people who use them in food preparation.