Leishmania-specific isotype levels and their relationship with specific cell-mediated immunity parameters in canine leishmaniasis

In the current retrospective study, Leishmania infantum-specific IgG, IgA and IgM levels were determined by ELISA in 106 untreated dogs with clinically-patent leishmaniasis (Sx) and in 171 clinically healthy dogs (Asx) from Spain to investigate the relationship between these Ig isotypes and clinical status. In addition, we studied if different Leishmania-specific humoral pattern exists between Asx dogs with and without cellular mediated immunity (CMI). Fifty-six dogs were assessed by means of lymphoproliferation assay (LPA), interferon production (IFN) and leishmanin skin test (LST), 71 dogs by means of both LPA and IFN and 44 only by LST. Both Sx and Asx dogs produced Leishmania-specific IgG, IgA and IgM antibodies, however the levels and proportion of positive dogs for each Ig isotype were significantly higher in Sx than in Asx ones (P<0.001). Analysis of immunological profiles determined for each cellular technique (positive and negative cellular response for each technique combined with positive or negative specific humoral response) showed that Asx dogs constituted a high heterogeneous group. No correlations were observed between CMI tests and specific IgG or IgM levels. However, a significant inverse correlation was demonstrated between specific IgA levels and LPA response (Spearman's r=-0.220; P=0.035). In general, the low correlation detected between CMI tests and isotype levels might indicate that the immune response is not strongly polarized in the majority of Asx dogs. Additionally, this study suggests that dogs developing T-cell response are probably able to avoid the dissemination of the parasite at least to mucosal surfaces and, as a consequence, to produce low or background specific IgA levels. Further studies are needed to investigate the relationship between specific IgA and parasite load, especially at mucosal site.     

Study of some Saccharomyces cerevisiae strains for winemaking after preadaptation at low temperatures

Low-temperature fermentations (13 degrees C) are considered to improve wine aromatic profiles. However, because the risk of stuck and sluggish fermentations is high, these fermentations are not common. The aim of this paper was to analyze the effect of different preadaptation protocols in two commercial wine strains on the fermentation and some wine parameters. Preadaptation is understood to be the process between the rehydration of active dry yeast and the inoculation. In this study, it consisted of preparing a fermentation starter (addition of yeast grown at 25 degrees C) or inocula preadapted at low temperatures (as before, but grown at a fermentation temperature of 13 or 17 degrees C). These results were compared with those of rehydrated active dry yeast, and a commercial "cryotolerant" yeast was used as a reference. General fermentation kinetic parameters, yeast imposition, nitrogen consumption, and main wine products were analyzed. The results showed that the preadaptation of a yeast could improve the fermentation performance, although this improvement was strain-dependent. Low-temperature fermentations also had some general effects: reduction of acetic acid and fusel alcohol production and increased concentrations of glycerol. When the yeast performed better in fermentation because of preadaptation, nitrogen consumption was faster and the wine's "negative" attributes (acetic acid, fusel alcohols) were significantly reduced. Thus, in some strains, preadaptation could be an effective mechanism for improving low-temperature fermentation, which also significantly reduces detrimental wine attributes.     

Isolation of 4-methylthio-3-butenyl glucosinolate from Raphanus sativus sprouts (kaiware daikon) and its redox properties

The most promising among glucosinolates (GLs) are those bearing in their aglycon an extra sulfur function, such as glucoraphasatin (4-methylthio-3-butenyl GL; GRH) and glucoraphenin (4-methylsulfinyl-3-butenyl GL; GRE). The GRE/GRH redox couple is typically met among secondary metabolites of Raphanus sativus L. and, whereas GRE prevails in seeds, GRH is the major GL in full-grown roots. During the 10 days of sprouting of R. sativus seeds, the GRE and GRH contents were determined according to the Eurpean Union official method (ISO 9167-1). In comparison to the seeds, the GRE content in sprouts decreased from about 90 to about 12 micromol g(-1) of dry weight (dw), whereas a 25-fold increase--from about 3 to 76 micromol g(-1) of dw--of the GRH content was measured. An efficient pure GRH gram-scale production process from R. sativus (kaiware daikon) sprouts resulted in significant yield improvement of up to 2.2% (dw basis). The reaction of GRH with both H2O2 and ABTS*+ radical cation was investigated. Whereas H2O2 oxidation of GRH readily resulted in complete transformation into GRE, ABTS*+ caused complete decay of the GL. Even though not directly related to its radical scavenging activity, the assessed reducing capacity of GRH suggests that R. sativus sprouts might possess potential for health benefits.     

Effect of calcium on the oxidation of linoleic acid by potato (Solanum tuberosum var. Desiree) tuber 5-lipoxygenase

When the effect of calcium on the oxidation of linoleic acid by potato tuber 5-lipoxygenase (LOX) was investigated, it was seen to promote the enzyme's activity at pH values higher than the optimum pH of 6.3, resulting in an enzyme activation at alkaline pH. Kinetic analysis of calcium activation at different pH values revealed that the cation abolished the inhibition by high substrate concentration, which occurs in the absence of Ca(2+), thus leading to activation at high substrate concentration. Studies were conducted to investigate the influence of Ca(2+) on the physicochemical nature of the substrate and its effect on the LOX activity expression. It was concluded that the aggregation mode rather than the aggregation state of linoleic acid is responsible for potato 5-LOX changes.     

Rapid surface plasmon resonance-based inhibition assay of deoxynivalenol

Deoxynivalenol belongs to a group of highly toxic fungal metabolites produced by Fusarium species that may contaminate food and animal feed, mostly grains. Three different monoclonal mouse anti-deoxynivalenol antibodies were compared for the development of a surface plasmon resonance (SPR)-based immunoassay for the selective and quantitative determination of deoxynivalenol in naturally contaminated matrices. A conjugate of deoxynivalenol with the protein casein was prepared and immobilized on the sensor chip surface. An excess of antibody was added to each test solution before the measurement. The assay was based on the competition for antibody binding between the immobilized deoxynivalenol conjugate on the sensor and the free deoxynivalenol molecules in the test solution. The deoxynivalenol-casein sensor could be reused more than 500 times without significant loss of activity using 6 M guanidine chloride solution for regeneration. The cross-reactivity of the three antibodies in the SPR assay was tested with other trichothecene mycotoxins (3-acetyl-deoxynivalenol, 15-acetyl-deoxynivalenol, nivalenol, HT2-toxin, and T2-toxin). The only sample preparation was extraction with max 80 vol % acetonitrile and 10-fold dilution with the running buffer. The assay had an optimal range between 2.5 and 30 ng/mL deoxynivalenol in the test solution. Most results of the SPR-based assay were in agreement with liquid chromatography/tandem mass spectrometry measurements of naturally contaminated wheat samples.     

Identification and quantification of carotenoids including geometrical isomers in fruit and vegetable juices by liquid chromatography with ultraviolet-diode array detection

A method was established for the identification and quantification of carotenoids including geometrical isomers in fruit and vegetable juices by liquid chromatography with an ultraviolet-diode array detector, using a C(18) Vydac 201TP54 column. The mobile phase used was the ternary methanol mixture (0.1 M ammonium acetate), tert-butyl methyl ether and water, in a concentration gradient, and a temperature gradient was applied. Retinol palmitate was added as an internal standard. An extraction process (ethanol/hexane, 4:3, v/v) was performed, followed by saponification with diethyl ether/methanolic KOH (0.1%, w/v, BHT) (1:1, v/v) for 0.5 h at room temperature. Seventeen different (cis and trans) carotenoids were identified by UV-vis spectra and retention times in HPLC in the juices analyzed. The analytic parameters show that the method proposed is sensitive, reliable, accurate, and reproducible.     

Polyphenol oxidase from Dominga table grape

Dominga grape polyphenol oxidase (PPO) was extracted using phase partitioning with Triton X-114. The enzyme was obtained in latent state and could be optimally activated by the presence of 0.2% sodium dodecyl sulfate (SDS) at pH 6.0. In the absence of SDS, the enzyme showed maximum activity at acid pH. The kinetic parameters of the enzyme at pH 3.0 and 6.0 in the presence of SDS were calculated. The effect of several inhibitors was studied, tropolone being the most effective with a K(i) value of 18 muM. The effect of cyclodextrins was also studied, and the complexation constant K(c) between G(2)-beta-cyclodextrins and 4-tert-butylcatechol was calculated using the enzymatic method (K(c) = 13960 M(-)(1)). The evolution of the color parameters (L, a, b) of liquefied grape berries was inhibited by inhibitors of PPO activity, such as diethyldithiocarbamate, metabisulfite, and G(2)-beta-cyclodextrins, indicating that enzymatic browning by PPO is the main process involved in the browning of Dominga grape juice at room temperature.     

Prediction chemical composition and alveograph parameters on wheat by near-infrared transmittance spectroscopy

Moisture, protein, wet gluten, dry gluten, and alveograph parameters (W, P, and P/L) of whole wheat grown in different countries around the world were analyzed using near infrared (NIR) transmittance spectroscopy. Modified partial least squares on NIR spectra (850-1048.2 nm) were developed for each constituent or physical property. The best models were obtained for protein, moisture, wet gluten, and dry gluten with r(2) = 0.99, 0.99, 0.95, and 0.96, respectively. Initial alveograph NIR models proposed for all wheat samples did not perform well. However, when wheat samples were divided in two groups depending on W (deformation energy) values, NIR models were highly improved, showing enough prediction accuracy for screening wheat at the receiving stage at mills or elevators.     

Different tocopherols and the relationship between two methods for determination of primary oxidation products in fish oil

The effectiveness of 2.32 mmol/kg (approximately 1000 ppm) of alpha-, gamma-, or delta-tocopherol (TOH), as well as different levels of alpha TOH, on the formation of hydroperoxides in fish oil was studied by monitoring the peroxide value (POV) and the formation of conjugated dienes (CD) during storage at 30 degrees C. The same order of antioxidant activity was observed by both methods. Linear regression of POV on CD showed that these data were strongly correlated (r(2) > or = 0.98). The value of the slope of the regression lines, however, differed substantially and decreased with increasing hydrogen-donating ability of the different tocopherols and with increasing alpha TOH concentration. It is suggested that this is due, at least in part, to the contribution from hydroxy compounds to the CD measurements and a greater contribution from hydroperoxy epidioxides (two peroxide groups per conjugated diene unit) to the POV than to the CD value. The degrees of formation of both these groups of oxidation products are expected to be influenced by the rate of scavenging of lipid peroxyl and alkoxyl radicals by tocopherol (alpha TOH > gamma TOH > delta TOH).     

钙盐和热处理对鲜切“Galia”甜瓜品质保鲜效果的研究

"Galia"甜瓜是西班牙主要的鲜食及加工用甜瓜品种,然而,鲜切甜瓜在贮藏过程中非常容易软化,并且在气调贮藏时容易受到冷害。其软化的过程和果实组织中钙(Ca)元素的含量水平有关。试验将切成梯形的"Galia"甜瓜果实分别在Ca浓度为0.4%(0.15 g.g-1),温度为60℃的氯化物、柠檬酸盐、乳酸盐、抗坏血酸盐、酒石酸盐、硅酸盐、丙酸盐和醋酸盐溶液中浸泡1min,采用50mg.L-1的H2O2控制微生物的生长,以60℃无菌水(无钙盐)中浸泡1min为对照。在5℃、4.5 kPa的O2和14.7 kPa的CO2的气调环境下贮藏10 d,测定硬度、果胶甲酯酶和多聚半乳糖醛酸酶活性、Ca含量、微生物生长量、呼吸速率和感官品质指标。在货架期结束时,抗坏血酸钙、氯化钙和乳酸钙处理的鲜切甜瓜果实呼吸速率较低,组织中的Ca含量增加,硬度较好,同时,这三种Ca盐处理还抑制了微生物的生长,鲜切甜瓜的感官品质(如风味水平)优于市场需求的水平,得到消费者的认可,而其他处理的风味均有较大幅度地下降。