Functional stabilization of cellulase by covalent modification with chitosan

Chitosan was linked to cellulase (EC 3.2.1.4) from Trichoderma viride by covalent conjugation to periodate‐activated carbohydrate moieties of the enzyme. The modified enzyme contained about 1.5 mol of polymer per mol of protein. The specific activity of the conjugate prepared was 39.8% of the native cellulase. The optimum pH and temperature for cellulase remained unaltered after modification. The thermostability was increased by 8.9 °C for the cellulase–chitosan complex. Thermal inactivation at different temperatures ranging from 65 °C to 80 °C was markedly increased for the polymer‐modified enzyme. The stability within the pH range 1.0–3.2 was also improved for the modified enzyme. © 2001 Society of Chemical Industry     

Incorporation and distribution of saturated and unsaturated fatty acids into nuclear lipids of hepatic cells

Liver nuclear incorporation of stearic (18∶0), linoleic (18∶2n−6), and arachidonic (20∶4n−6) acids was studied by incubation in vitro of the [1-¹⁴C] fatty acids with nuclei, with or without the cytosol fraction at different times. The [1-¹⁴C] fatty acids were incorporated into the nuclei as free fatty acids in the following order: 18∶0>20∶4n−6≫18∶2n−6, and esterified into nuclear lipids by an acyl-CoA pathway. All [1-¹⁴C] fatty acids were esterified mainly to phospholipids and triacylglycerols and in a minor proportion to diacylglycerols. Only [1-¹⁴C] 18∶2n−6-CoA was incorporated into cholesterol esters. The incorporation was not modified by cytosol addition. The incorporation of 20∶4n−6 into nuclear phosphatidylcholine (PC) pools was also studied by incubation of liver nuclei in vitro with [1-¹⁴C]20∶4n−6-CoA, and nuclear labeled PC molecular species were determined. From the 15 PC nuclear molecular species determined, five were labeled with [1-¹⁴C]20∶4n−6-CoA: 18∶0–20∶4, 16∶0–20∶4, 18∶1–20∶4, 18∶2–20∶4, and 20∶4–20∶4. The highest specific radioactivity was found in 20∶4–20∶4 PC, which is a minor species. In conclusion, liver cell nuclei possess the necessary enzymes to incorporate exogenous saturated and unsaturated fatty acids into lipids by an acyl-CoA pathway, showing specificity for each fatty acid. Liver cell nuclei also utilize exogenous 20∶4n−6-CoA to synthesize the major molecular species of PC with 20∶4n−6 at the sn-2 position. However, the most actively synthesized is 20∶4–20∶4 PC, which is a quantitatively minor component. The labeling pattern of 20∶4–20∶4 PC would indicate that this molecular species is synthesized mainly by the de novo pathway.     

An adaptive color similarity function suitable for image segmentation and its numerical evaluation

In this article, we present an adaptive color similarity function defined in a modified hue‐saturation‐intensity color space, which can be used directly as a metric to obtain pixel‐wise segmentation of color images among other applications. The color information of every pixel is integrated as a unit by an adaptive similarity function thus avoiding color information scattering. As a direct application we present an efficient interactive, supervised color segmentation method with linear complexity respect to the number of pixels of the input image. The process has three steps: (1) Manual selection of few pixels in a sample of the color to be segmented. (2) Automatic generation of the so called color similarity image (CSI), which is a gray level image with all the gray level tonalities associated with the selected color. (3) Automatic threshold of the CSI to obtain the final segmentation. The proposed technique is direct, simple and computationally inexpensive. The evaluation of the efficiency of the color segmentation method is presented showing good performance in all cases of study. A comparative study is made between the behavior of the proposed method and two comparable segmentation techniques in color images using (1) the Euclidean metric of the a* and b* color channels rejecting L* and (2) a probabilistic approach on a* and b* in the CIE L*a*b* color space. Our testing system can be used either to explore the behavior of a similarity function (or metric) in different color spaces or to explore different metrics (or similarity functions) in the same color space. It was obtained from the results that the color parameters a* and b* are not independent of the luminance parameter L* as one might initially assume in the CIE L*a*b* color space. We show that our solution improves the quality of the proposed color segmentation technique and its quick result is significant with respect to other solutions found in the literature. The method also gives a good performance in low chromaticity, gray level and low contrast images. © 2016 Wiley Periodicals, Inc. Col Res Appl, 42, 156–172, 2017     

Study and comparison of different drying processes for dehydration of raspberries

The aim of the present study was to evaluate and compare different drying methods (microwave, hot air?+?microwave, and osmotic dehydration?+?microwave) in raspberries (cv. Heritage). A portion of raspberries was pretreated with osmotic dehydration (60°Brix sucrose solution at 20°C for 360?min) and another with hot air drying (HAD) (1.5?m/s air speed at 60°C for 300?min). Pretreated raspberries were then dried by microwave and at three different intensities (3.5, 7.5, and 11?W/g). Physicochemical properties (moisture content, water activity, and drying rate) and quality parameters (optical properties, mechanical properties, antioxidant capacity, and rehydration capacity) of dried raspberries were evaluated. Results showed that the microwave drying (MWD) at 7.5?W/g (50?min and final temperature of 79?±?5.1°C) allowed a high yield of dried raspberries. The combined processes were not efficient to accelerate the decrease of moisture content, due to the low drying rate of the pretreatments. In terms of quality, none of the drying processes allowed a high retention of the antioxidant capacity. However, they allowed an appropriate rehydration performance. The combination of HAD with MWD allowed obtaining a good appearance and desirable texture on the dried product. Thus, this last option seems to be the best among the drying methods tested, but additional studies are required to improve the efficiency of the process and the effect on the antioxidant capacity during drying.     

Maximum volumetric production of β‐galactosidase by Kluyveromyces fragilis in fed‐batch culture with automatic control

The production of β‐galactosidase by Kluyveromyces fragilis was studied in different culture systems, with dissolved oxygen concentration control and using defined media. An operating strategy of fed‐batch culture with automatic control of substrate addition regulated by dissolved oxygen concentration, consisting of the replacement of variable volumes of broth by fresh medium (once the fed‐batch culture has finished), was designed. The volumetric enzyme productivity (Q_p, 13 600 UI dm^?3 h^?1) obtained was 38% higher than that reached in continuous culture of K fragilis with dissolved oxygen concentration control and far higher than that obtained by batch culture of K fragilis under the same aeration conditions. © 2002 Society of Chemical Industry     

Up-Regulation of mRNA Ventricular PRNP Prion Protein Gene Expression in Air Pollution Highly Exposed Young Urbanites: Endoplasmic Reticulum Stress,Glucose Regulated Protein 78, and Nanosized Particles

Mexico City Metropolitan Area children and young adults exposed to high concentrations of air pollutants including fine and ultrafine particulate matter (PM) vs. clean air controls, exhibit myocardial inflammation and inflammasome activation with a differential right and left ventricular expression of key inflammatory genes and inflammasomes. We investigated the mRNA expression levels of the prion protein gene PRNP, which plays an important role in the protection against oxidative stress and metal toxicity, and the glucose regulated protein 78, a key protein in endoplasmic reticulum (ER) stress signaling, in ventricular autopsy samples from 30 children and young adults age 19.97 ± 6.8 years with a lifetime of low (n:4) vs. high (n:26) air pollution exposures. Light microscopy and transmission electron microscopy studies were carried out in human ventricles, and electron microscopy studies were also done in 5 young, highly exposed Mexico City dogs. There was significant left ventricular PRNP and bi-ventricular GRP78 mRNA up-regulation in Mexico City young urbanites vs. controls. PRNP up-regulation in the left ventricle was significantly different from the right, p < 0.0001, and there was a strong left ventricular PRNP and GRP78 correlation (p = 0.0005). Marked abnormalities in capillary endothelial cells, numerous nanosized particles in myocardial ER and in abnormal mitochondria characterized the highly exposed ventricles. Early and sustained cardiac ER stress could result in detrimental irreversible consequences in urban children, and while highly complex systems maintain myocardial homeostasis, failure to compensate for chronic myocardial inflammation, oxidative and ER stress, and particles damaging myocardial organelles may prime the development of pathophysiological cardiovascular states in young urbanites. Nanosized PM could play a key cardiac myocyte toxicity role.     

Application of titanium machining chips in welding consumables for wear-resistant hardfacing

Equipment of sugar cane plants and mineral extraction are submitted to severe abrasive wear conditions. Welded hardfacings are usually applied to repair this kind of damage, where commercial chromium/carbon-rich welding consumables have usually been employed. In the present work we investigated the microstructure of experimental hardfacings made by addition of residues (chips) collected from the machining of ASTM F67 (unalloyed Ti, grade 4) alloy. Mixtures with different carbide-formers (Cr/Nb ferro-alloys) were also tested. Two layers of ‘pure’ chips (Ti), chips plus Fe–Cr (Ti–Cr), and chips plus Fe–Nb (Ti–Nb) were applied on low-carbon steel specimens by the GTAW/TIG process. The microstructure of hardfacing layers was observed by optical and scanning electron microscopy (SEM) equipped with EDS microanalysis. The microstructural characterization has determined that carbide distributions change significantly with the chemical nature of the hardfacing. SEM observations coupled with EDS microanalysis have confirmed the formation of complex carbides within the metal weld, whose stoichiometry was determined by X-ray diffraction (XRD) analysis. Mixed carbides of MC type and some cementite have been found. As a result it was suggested that use of ASTM F67 chips as carbide formers for composition of welding consumables can contribute to improved wear resistance of hardfacings, if compared with traditional chromium-based hardfacings.     

Fatty acid Δ9-Desaturation in the Triatoma infestans fat body: Response to food and trehalose administrationsin the Triatoma infestans fat body: Response to food and trehalose administrations

The effects of food intake and carbohydrate administration on fatty acid Δ⁹-desaturation were investigated in isolated microsomes from Triatoma infestans fat body. Fifth instar nymphs, which were solely blood-fed just after the molt and then fasted, were used as controls and for determination of optimal assay conditions. Both [1-¹⁴C]palmitic and [1-¹⁴C]stearic acids in a medium containing ATP, CoA, MgCl₂, NADH, NaF, and O₂ were tested. For the control group, optimal conditions were a pH of 6.8–7.2, an incubation temperature of 29°C, and an incubation time of 10 min. Δ⁹-desaturation depended on the presence of reduced pyridine dinucleotides; NADH and NADPH were equally efficient. Stearic acid showed a higher apparent V _max than palmitic acid, but the apparent K _m were very similar. When fifth instar nymphs were blood-fed weekly, a marked increase of Δ⁹-desaturation was observed for both acids. Higher desaturation activity was also induced by injection of the carbohydrate trehalose into the hemolymph of fasted nymphs. These results indicate that insect Δ⁹-desaturation, like the mammalian counterpart, is sensitive to dietary changes and carbohydrate administration. *** DIRECT SUPPORT *** A01K6035 00008     

Effect of modified ITO substrate on electrochromic properties of polyaniline films

In this work, we report the morphological and electrochromic properties of electrochemically synthesized polyaniline (PANI) thin films on bare and modified indium–tin oxide (ITO) glass substrates. In the last case, the surface of ITO glass was covered by a self-assembled monolayer of N-phenyl-γ-aminopropyl-trimethoxysilane (PAPTS). Atomic force microscopy images and perfilometry show that smoother and thinner PANI films were grown on PAPTS-modified ITO substrates. PANI-based electrochromic devices (ECDs) were assembled by using a viscous polymeric electrolyte (PE) of LiClO₄ and polymethyl methacrylate (PMMA) co-dissolved in a mixture of propylene and ethylene carbonate. The architectural design of the devices was glass/ITO/PANI/PE/ITO/glass. A dual ECD was also prepared by collocating a poly(3-methylthiophene) (P3MT) thin film as a complementary electrochromic element. The effect of the PAPTS-modified ITO substrate is reflected in a higher optical transmittance at bleach state and a little less color change at 550 nm of PANI-based ECDs.