Cysteine proteinase inhibitor Act d 4 is a functional allergen contributing to the clinical symptoms of kiwifruit allergy

Kiwifruit has become a frequent cause of fruit allergy in the recent years. The molecular basis of type I hypersensitivity to kiwifruit is attributed to 11 IUIS allergens, with Act d 1, Act d 2 and Act d 5 characterized in extenso. Evaluation of the allergenic properties of Act d 4, a cysteine proteinase inhibitor from green kiwifruit (Actinidia deliciosa) was performed in this study. Identity of the purified glycoprotein was determined by Edman degradation and by mass fingerprint whereby more than 90% of the primary structure of the mature kiwifruit cystatin was confirmed. Using MALDI TOF analysis, molecular masses of 10902.5 and 11055.2 Da were detected for Act d 4, respectively. Positive skin prick reactivity with Act d 4 was induced in three kiwifruit allergic patients, as well as the upregulation of CD63 and CD203c molecules in the basophile activation assay. The IgE reactivity was detected in dot blot analysis while Western blot analysis was negative using sera from six kiwifruit patients, suggesting the presence of conformational IgE epitopes on the Act d 4 molecule. As activator of effector cells in type I hypersensitivity Act d 4 is a functional allergen contributing to the clinical symptoms of kiwifruit allergy.     

Macromolecular Changes in Extruded Starch‐Films Plasticized with Glycerol,Water and Stearic Acid

Native corn starch, plasticized with water, glycerol and stearic acid, was extruded in a conical twin‐screw extruder and sheeted into 0.4–0.6 mm thick films. The effects of extrusion and plasticizers on gelatinization, as well as the molecular and structural changes, in thermoplastic starch were analyzed. The onset and peak gelatinization temperatures of extruded starch varied from 42–46°C and 52.9–56.9°C, respectively, depending on the glycerol content. The enthalpy of gelatinization of extruded thermoplastic starch in excess water varied from 3.6–7.6 J/g, which also increased with plasticizer content. Amylose‐lipid complexes were formed during extrusion, and their enthalpies depended on the initial stearic acid and moisture contents. High‐performance size‐exclusion chromatography (HPSEC) data revealed that the starch underwent fragmentation during extrusion even under highly plasticized conditions, but the degradation was not severe as compared to previous findings. The relative percentages of amylopectin and amylose in native starch were 76.9 and 23.1%, respectively, which were changed to 71.3–76.6% and 23.4–28.7% in the extrudates. The average molecular weights of amylopectin and amylose in the extrudates ranged from 1.55×10⁷–2.07×10⁷ and 4.35×10⁵–7.39×10⁵, respectively. On the other hand, the molecular weights of amylopectin and amylose in native corn starch were observed as 2.27×10⁷ and 4.68×10⁵, respectively. Cross‐polarization magical angle spinning (CP/MAS) and high‐power decoupling (HP‐DEC) nuclear magnetic resonance (NMR) spectra of thermoplastic starch revealed the characteristics of amylomaize starch, confirming HPSEC results that the amylopectin macromolecules underwent fragmentation into amylose‐like fractions. In the extrudates, glycerol was found to be less mobile and entrained within the starch network.     

Solvothermal Synthesis and Catalytic Properties of Nanocrystalline ZnFe<Subscript>2−x</Subscript>Al<Subscript>x</Subscript>O<Subscript>4</Subscript> (x = 0, 1, 2) Spinels in Aniline Methylation

Abstract  

ZnFe_2−xAl_xO₄ (x = 0, 1, 2) spinels were obtained by microwave-assisted solvothermal method using 1,4-butanediol as reaction medium. The results of XRD and HRTEM studies have indicated higher nanocrystallinity of aluminium containing spinels, and N₂ adsorption–desorption measurements have revealed their enhanced textural properties, in particular much higher specific surface areas. NH₃-TPD method and cyclohexanol test have shown the variation of surface acid–base properties with the changes in the spinel composition. All studied spinels were active in aniline methylation and proved to be selective for N-methylation leading to N-methylaniline and N,N-dimethylaniline. The main advantage of aluminium containing spinels is that N-alkylation with methanol is possible at significantly lower temperature (200–260 °C) when pure zinc ferrite is almost inactive.     

Transport of two naphthoic acids and salicylic acid in soil: experimental study and empirical modeling

In contrast to the parent compounds, the mechanisms responsible for the transport of natural metabolites of polycyclic aromatic hydrocarbons (PAH) in contaminated soils have been scarcely investigated. In this study, the sorption of three aromatic acids (1-naphthoic acid (NA), 1-hydroxy-2-naphthoic acid (HNA) and salicylic acid (SA)) was examined on soil, in a batch equilibrium single-system, with varying pH and acid concentrations. Continuous flow experiments were also carried out under steady-state water flow. The adsorption behavior of naphthoic and benzoic acids was affected by ligand functionality and molecular structure. All modeling options (equilibrium, chemical nonequilibrium, i.e. chemical kinetics, physical nonequilibrium, i.e. surface sites in the immobile water fraction, and both chemical and physical nonequilibrium) were tested in order to describe the breakthrough behavior of organic compounds in homogeneously packed soil columns. Tracer experiments showed a small fractionation of flow into mobile and immobile compartments, and the related hydrodynamic parameters were used for the modeling of reactive transport. In all cases, the isotherm parameters obtained from column tests differed from those derived from the batch experiments. The best accurate modeling was obtained considering nonequilibrium for the three organic compounds. Both chemical and physical nonequilibrium led to appropriate modeling for HNA and NA, while chemical nonequilibrium was the sole option for SA. SA sorption occurs mainly in mobile water and results from the concomitancy of instantaneous and kinetically limited sites. For all organic compounds, retention is contact condition dependent and differs between batch and column experiments. Such results show that preponderant mechanisms are solute dependent and kinetically limited, which has important implications for the fate and transport of carboxylated aromatic compounds in contaminated soils.     

Purification and characterisation of angiotensin I converting enzyme inhibitory peptides from lysozyme hydrolysates

Hen egg white lysozyme (HEWL) was hydrolysed with trypsin, papain and a combination of the two. The prepared hydrolysates exhibited ACE inhibitory activity. The hydrolysates were fractionated using ultrafiltration and reverse phase-high performance liquid chromatography (RP-HPLC). Three fractions, which showed the highest ACE inhibitory activities, were purified by RP-HPLC. They were the F₇ (from papain-trypsin hydrolysate), F₈ (from papain hydrolysate) and F₃ (from trypsin hydrolysate) fractions. The IC₅₀ values were 0.03, 0.155 and 0.23 mg/ml for F₇, F₈ and F₃, respectively. The F₇ fraction was the most potent ACE inhibitor peptide, and was composed of 12 amino acids, Phe-Glu-Ser-Asn-Phe-Asn-Thr-Gln-Ala-Thr-Asn-Arg (MW: 1428.6 Da). Lineweaver-Burk plots suggest that the F₇ peptide acts as an uncompetitive inhibitor against ACE. The kinetic parameters (K_m, V_max, and K_i) for the F₇ peptide were measured and compared to the control.     

Bactericidal effects of lettuce after subsequent washing with hypobromous acid and sodium hypochlorite

Sodium hypochlorite (NaClO) is widely used as a sanitizer to maintain microbial safety of lettuce; however, NaClO treatment is insufficient on pathogen reduction due to its high instability for an extended storage time. Hypobromous acid (HOBr) as another sanitizer, relatively stable in processing conditions, was applied to lettuce. The numbers of total aerobic bacteria, Escherichia coli, Salmonella Typhimurium, and Staphylococcus aureus were evaluated on lettuce treated with NaClO and HOBr. The population of 4 selected bacteria on lettuce reduced more than 2.5 log CFU/g. Bactericidal effect maintained for 10 days in combined treatment with 100 mg/L NaClO followed by 200 mg/L HOBr, while the population of the bacteria without HOBr treatment increased. Combined treatment on lettuce can give synergistic effect on reducing microbial population. Residual bromine was not detected on the lettuce treated with HOBr after washed with water for 30 s, indicating that bromine was effectively washed away.     

Enhancing antioxidant availability in wheat grains from plants grown under seawater stress in response to microalgae extract treatments

BACKGROUND: The present study was designed to investigate the enhanced antioxidant capacity of whole grains from wheat plants grown under seawater stress in response to microalgae extract treatment. RESULTS: The total carotenoid (TCAR), tocopherol (TOC), phenolic (TPC), and protein (PC) contents in whole grains of wheat plants irrigated with 10% and 20% (v/v) seawater (SW) in response to water extracts of microalgae Spirulina maxima (SME) and Chlorella ellipsoida (CEE) as well as exogenous plant growth enhancers of ascorbic acid and benzyladenine treatments were measured. The results showed that the levels of enhanced TCAR (range 0.08–0.14 g kg^?1), TOC (range 0.05–0.12 g kg^?1), TPC (range 0.80–2.96 g kg^?1) and PC (range 93.4–137.9 g kg^?1) in wheat grains of plants irrigated with 10% and 20% SW were significantly increased in response to SME and CEE treatments. Evaluation of antioxidant activity of ethanolic extracts of grains of SW‐stressed plants indicated that DPPH and TBAS radical scavenging activity was significantly increased in response to SME and CEE treatments and coincided with the increase in levels of antioxidant compounds present in each extract. The electrophoretic profiles of the grains of proteins of treated samples exhibited quite different patterns from those in control samples. CONCLUSION: The results suggest that the application of algal extracts to wheat plants irrigated with SW is useful for improvement of salinity tolerance. This effect can be triggered by the stimulation of antioxidant components and protein content. Copyright © 2009 Society of Chemical Industry     

Nitrate modifies the assimilation pattern of ammonium and urea in wheat seedlings

BACKGROUND: In certain plant species, ammonium or urea nutrition can cause negative effects on plant development which can result in toxic symptoms. Some authors suggest that the presence of nitrate can alleviate these symptoms by increasing ammonium and urea assimilation, avoiding its accumulation. In order to study this hypothesis, wheat (Triticum aestivum L.) seedlings were grown with various nitrogen supplies containing the main nitrogen forms (ammonium, nitrate and urea). Amino acids content and the activity of the three main enzymes involved in nitrogen assimilation (nitrate reductase, glutamine synthetase and urease) were studied. RESULTS: The application of nitrate along with urea and/or ammonium was not associated with a time‐sustained increase in the activity of glutamine synthetase and urease. Amino acid analysis revealed that nitrate induced changes in amino acid metabolism enhancing its concentration. Likewise the content of protein was also higher in nitrate‐treated plants. CONCLUSION: These results suggest that the effect of nitrate is compatible with a rapid and transient increase in the activity of glutamine synthetase and urease during the first hour after the onset of treatments. Nevertheless, a possible effect of nitrate reducing ammonium accumulation through the activation of alternative metabolic pathways different from that involving glutamine synthetase cannot be ruled out. Finally, nitrate effects on amino acid concentration indicate that, whereas ammonium assimilation takes place principally in the root, urea and nitrate assimilation occurred in the shoot, under the conditions of the experiment. Copyright © 2009 Society of Chemical Industry     

Left ventricular morphology and function in diabetic and nondiabetic hemodialyzed patients

Morbidity in end-stage renal disease (ESRD) diabetic patients is worse than in patients without diabetes mellitus (DM). This study aims to compare clinical, laboratory, and echocardiographic features between the ESRD patients with and without DM. Fifty-eight ESRD patients on dialysis were prospectively divided into two groups according to the presence of DM. Demographic, clinical, laboratory, and echocardiographic features (ejection fraction and wall motion score index) were compared between the two groups. Overall, 20 out of 58 patients (37.8%) with ESRD had DM. There were no significant differences between the patients with DM and those without DM when it comes to age (60.6 ± 10.6 vs. 59.0 ± 10.6 years, P = 0.665), ejection fraction (52.6% ± 12.8% vs. 54.2% ± 12.8%, P = 0.59), and wall motion score index (1.21 ± 0.3 vs.1.15 ± 0.3, P = 0.37). In multivariant analysis of the interventricular septum, posterior wall thickness and left atrium size correlated positively with DM. There was also no statistical difference in myocardial perfusion disturbances on real-time contrast echocardiography between the groups with and without DM (12 (60%) patients vs. 14 patients (36.8%), P = 0.079). Among diabetics 77.8% had significant atherosclerotic changes, while in the group without DM, only 38.1%, P = 0.01. From the laboratory parameters ferritin and high-sensitivity C-reactive protein levels were significantly higher in the group with DM, P = 0.014 and P = 0.026, respectively. Patients with ESRD and DM have significantly bigger left atrial size, thicker left ventricular walls, and higher serum ferritin and high-sensitivity C-reactive protein levels than the patients without DM. The aforementioned features may be possible risk factors for the development of adverse cardiac events in patients on hemodialysis.