Impact of Maillard reaction on immunoreactivity and allergenicity of the hazelnut allergen Cor a 11

Few studies exist on the influence of processing methods on structural changes and allergenic potential of hazelnut proteins. This study focused on the effect of glycation (Maillard reaction) on the immunoreactivity and degranulation capacity of the purified hazelnut 7S globulin, Cor a 11. After heating, the extent of the Maillard reaction, sensitivity to proteolysis, binding of human IgE or rabbit IgG, and degranulation capacity were analyzed. Changes in electrophoretic mobility, amount of free amino groups, and contents of bound sugar and fructosamine indicated that glycation of Cor a 11 occurred at all conditions. Glycation at 37 °C did not influence the specific IgG or IgE binding and was decreased after heating at 60 and 145 °C. Heating, with or without glucose, at 145 °C increased basophil degranulation capacity. The results suggest that glycation of Cor a 11 at 60 and 145 °C may decrease the IgE/IgG binding properties but not the degranulation capacity of basophils. This is possibly related to aggregation of the proteins as a result of the Maillard reaction.     

Patterns of ascospore release in relation to phoma stem canker epidemiology in England (<Emphasis Type="Italic">Leptosphaeria maculans</Emphasis>) and Poland (<Emphasis Type="Italic">Leptosphaeria biglobosa</Emphasis>)

Experiments over five growing seasons at Rothamsted (1998/99–2002/03), four seasons at Boxworth (1998/99, 1999/2000, 2001/02, 2002/03) in England (Leptosphaeria maculans) and three seasons (1998/99–2000/01) at Poznan in Poland (Leptosphaeria biglobosa) suggest that differences in the development of phoma stem canker epidemics between England and Poland relate to differences in weather patterns between the two countries. The duration of ascospore release was longer in England, where winter weather is mild and wet, than in Poland, where winters are cold and often with snow cover, but there was little difference between two sites in England (Rothamsted and Boxworth). Wetness provided by rainfall was essential for release of ascospores of both L. maculans in England and L. biglobosa in Poland. Temperature did not affect release of ascospores over the range 5–20 °C. Diurnal periodicity in release of ascospores of L. maculans in England and L. biglobosa in Poland was similar. The timing (date) of first release of ascospores of L. maculans or L. biglobosa in autumn was related to rainfall in August and September; with increasing rainfall the date was earlier. The incubation periods from first release of ascospores to first appearance of phoma leaf spots for both L. maculans in England and L. biglobosa in Poland, and from first leaf spots to first stem base canker in England, were described using a thermal time (degree-day) approximation.     

ASSESSMENT OF A RADIOTHERAPY PATIENT IMMOBILIZATION DEVICE USING SINGLE PLANE PORT RADIOGRAPHS AND A REMOTE COMPUTED TOMOGRAPHY SCANNER

Radiation treatment requires a precise procedure for interfraction repositioning of the patient. The purpose of this study was to determine the accuracy of our fixation device in treatment position and to evaluate the setup accuracy with two different methods. The positioning data of 19 canine patients with tumors in the head region (oral, nasal, cerebral) treated with photon or proton irradiation were included in this study. The patients were immobilized by means of an individualized fixation device. Focus was set upon interfraction displacement with systematic and random components. In one method, treatment position was evaluated using single plane port radiographs and megavoltage x-rays. In the other method, two orthogonal CT-topograms were acquired to evaluate the precision of positioning of the patient in the immobilization device. Systematic and random displacements were calculated and presented as mean values with corresponding 95% confidence intervals. In spite of a difference between both methods, the positioning seemed to be accurate within the expected range. It seems that a safety margin of 3.7 mm would be enough for both methods to take into account systematic and random position variability in the fixation device, thereby preventing geometric inaccuracies of treatment delivery. The reported immobilization protocol provides accurate patient immobilization for photon and conformal proton radiation therapy.     

Use of differential scanning calorimetry to study lipid oxidation. 1. Oxidative stability of lecithin and linolenic acid

The oxidation of linolenic acid (LNA) and soy lecithin was studied by differential scanning calorimetry (DSC) with linear programmed heating rates (non-isothermal mode). The interpretation of the shape of DSC curves is discussed, and it has been concluded that temperatures of the extrapolated start of heat release are the most reliable data for the rapid estimation of the oxidative stability of lipid materials. The Ozawa-Flynn-Wall method was used to calculate the kinetic parameters of the process: for LNA autoxidation the activation energy, Ea, and pre-exponential factor, Z, are 66 +/- 4 kJ/mol and 1.5 x 10(7) s(-1), respectively, and the autoxidation of lecithin is described by Ea = 98 +/- 6 kJ/mol and Z = 9.1 x 10(10) s(-1). Values of Ea and Z can be applied for calculation of the overall first-order rate constant of autoxidation at various temperatures, k(T). For the two studied lipids the comparison of k(T) values shows the inversion of their oxidative stabilities; that is, below 167 degrees C lecithin is more stable than LNA, k(T)lecithin < k(T)LNA, and above that temperature (termed the isokinetic temperature) k(T)lecithin > k(T)LNA. The calculated inversion of oxidative stabilities can be an explanation of similar observations for other pairs of lipids if the results of accelerated tests measured at temperatures above 100 degrees C are compared with the results obtained at temperatures below 100 degrees C.     

Nocardioform placentitis with isolation of Amycolatopsis spp in a Florida-bred mare

CASE DESCRIPTION: A 4-year-old Thoroughbred mare was evaluated because of placental abnormalities and a retained placental remnant. CLINICAL FINDINGS: Microbial culture of the placenta yielded pure growth of Amycolatopsis spp. Histologic examination of the placenta revealed a focally expanding chorionitis with intralesional gram-positive filamentous bacilli and multifocal allantoic adenomatous hyperplasia on the apposing allantoic surface. TREATMENT AND OUTCOME: Treatment with lavage and oxytocin resulted in expulsion of the placental remnant within hours of parturition. The mare did not become pregnant again despite multiple breedings. The foal appeared healthy but died of complications during an elective surgical procedure at 7 weeks of age. CONCLUSIONS AND CLINICAL RELEVANCE: To the author's knowledge, all previously confirmed cases of nocardioform placentitis have been in mares bred in the central Kentucky region. Indications that the pathogen in the mare reported here is a different species than that isolated in Kentucky suggest that this is an emerging disease. Mares with nocardioform placentitis usually do not have the same clinical signs as mares with placentitis resulting from an ascending pathogen.     

Effect of Diet Supplemented with Quinoa Seeds on Oxidative Status in Plasma and Selected Tissues of High Fructose-Fed Rats

Oxidative stress plays an important role as a mediator of damage produced by fructose metabolism. This work was designed to investigate the effect of diet supplemented with quinoa seeds on oxidative stress in plasma, heart, kidney, liver, spleen, lung, testis and pancreas of fructose administered rats. Fructose administration (310 g/kg fodder for 5 weeks) caused oxidative stress that was manifested by the increase in plasma malondialdehyde (MDA) (p<0.05), and by the non-significant changes in the enzymatic antioxidant potential in plasma and most of tissues. Co-administration of quinoa seeds (310 g/kg fodder) maintained normal activities of some enzymes. It also influenced the oxidative stress as was evidenced by decreasing MDA in plasma, and decreasing the activities of antioxidant enzymes (erythrocyte superoxide dismutase - eSOD, catalase -CAT, plasma glutathione peroxidase - pGPX). These findings demonstrate that quinoa seeds can act as a moderate protective agent against potential of fructose-induced changes in rats by reducing lipid peroxidation and by enhancing the antioxidant capacity of blood (plasma) and heart, kidney, testis, lung and pancreas.     

Effect of acetazolamide and subsequent ventriculo-peritoneal shunting on clinical signs and ventricular volumes in dogs with internal hydrocephalus

Background

Acetazolamide is recommended for the reduction of cerebrospinal fluid production in canine internal hydrocephalus. The efficacy of the drug in terms of alleviation of the clinical symptoms and the restoration of normal ventricular volume has not been documented. We hypothesize that acetazolamide inadequately improve clinical signs and has no effect on the ventricular volume. Six dogs with internal hydrocephalus underwent neurological examination and were examined by magnetic resonance imaging, on the day of the diagnosis, after treatment with acetazolamide directly before surgery, and 6 weeks after implantation of a vetriculo-peritoneal shunt due to lack of improvement after medical therapy with 10 mg/kg acetazolamide three times daily (TID). The ventricular volume in relation to the total brain volume was determined on each occasion. The changes in relative ventricular volume and of the neurological status were assessed and compared.

Results

McNemar’s test revealed no significant differences in clinical symptoms before and after medical treatment (P > 0.05). However, clinical symptoms changed significantly after surgical treatment (P = 0.001). The ventricle-brain ratio was not significantly changed after therapy with acetazolamide (P > 0.05); however, after subsequent shunt implantation, it was significantly reduced (P = 0.001).

Conclusion

Acetazolamide (10 mg/kg TID) showed no effects on clinical signs or ventricular volume in dogs with internal hydrocephalus. After subsequent ventriculo-peritoneal shunting, the dogs had a significantly reduced cerebral ventricular volume and five out of six dogs had no abnormal findings in neurological examination.     

The effect of winter weather conditions on the ability of pseudothecia of Leptosphaeria maculans and L. biglobosa to release ascospores

Leptosphaeria maculans and L. biglobosa are damaging pathogens of oilseed rape. The infection of plants occurs predominantly in early autumn or spring by spores produced in pseudothecia. The aim of this study was to investigate whether pseudothecia formed in the autumn are still viable in the spring and to what extend they are destroyed by winter frosts. The studies presented here demonstrated that winter frosts can render pseudothecia unable to release spores. Nevertheless, ascospores present in pseudothecia unable to discharge ascospores, were fully capable of germination, regardless of the incubation temperature. No significant differences were found between the studied Leptosphaeria species in their response to frost. A multiple regression equation has been elaborated to forecast the ability of pseudothecia to release ascospores, based on winter temperatures. Considerable correlation was found between the ascospore release in the autumn and the ability of pseudothecia to release ascospores over the winter period and the subsequent symptoms of stem canker before harvest. We have demonstrated that the potential and the survival of inoculum can have a large impact on the success of the pathogen. This may be particularly important in the light of forecasted climate change. Higher winter temperatures may increase the ability of pseudothecia to release ascospores and the discharge of ascospores of L. maculans and L. biglobosa into the air, and cause early plant infections. This in turn will increase the number of infected plants, the disease incidence at harvest, and reduce the yield of oilseed rape.     

Rep-PCR Based Genomic Fingerprinting of Isolates of Leptosphaeria maculans from Poland

Leptosphaeria maculans, the ascomycete fungus which causes blackleg disease of oilseed rape, has been considered for a long time as a single species divided into aggressive and non-aggressive pathogenicity groups which differ in their economic importance. However, the development of accurate biochemical and molecular characterisation methods has demonstrated that the world-wide L. maculans population actually comprises at least two species. The aim of this research was to assess the ability of rep (repetitive element based)-PCR genomic fingerprinting methods, initially developed for bacterial identification, to characterise a collection of 90 isolates of L. maculans from Poland, in comparison with reference isolates from the IBCN (International Blackleg of Crucifers Network) collection. REP (repetitive extragenic palindromic)-, ERIC (enterobacterial repetitive intergenic consensus)-, and BOX primers for rep-PCR genomic fingerprinting, and primers derived from LMR1, a L. maculans specific repeated element, were tested. Rep-PCR and LMR1-based analyses were able to discriminate the different components of the species complex and to evaluate the genetic diversity within each member of the complex. These analyses suggested that Polish populations of L. maculans mainly belong to the non-aggressive species, rather than the aggressive species which is prevalent in Western Europe, Canada and Australia.     

In situ simultaneous analysis of polyacetylenes, carotenoids and polysaccharides in carrot roots

This paper presents an approach to simultaneously analyze polyacetylenes, carotenoids, and polysaccharides in carrot (Daucus carota L.) roots by means of Raman spectroscopy. The components were measured in situ in the plant tissue without any preliminary sample preparation. The analysis is based on the intensive and characteristic key bands observed in the Raman spectrum of carrot root. The molecular structures of the main carrot polyacetylenes, falcarinol and falcarindiol, are similar, but their Raman spectra exhibit specific differences demonstrated by the shift of their -C[triple bond]C- mode from 2258 to 2252 cm(-)(1), respectively. Carotenoids can be identified by -C=C- stretching vibrations (about 1520 and 1155 cm(-)(1)) of the conjugated system of their polyene chain, whereas the characteristic Raman band at 478 cm(-)(1) indicates the skeletal vibration mode of starch molecule. The other polysaccharide, pectin, can be identified by the characteristic band at 854 cm(-)(1), which is due to the -C-O-C- skeletal mode of alpha-anomer carbohydrates. The Raman mapping technique applied here has revealed detailed information regarding the relative distribution of polyacetylenes, carotenoids, starch, and pectin in the investigated plant tissues. The distribution of these components varies among various carrot cultivars, and especially a significant difference can be seen between cultivated carrot and the wild relative D. carota ssp. maritimus.