We standardized and evaluated an ELISA technique for the detection of total and specific anti-Giardia duodenalis secretory IgA antibodies (sIgA). Samples of saliva and serum of 161 Venezuelan schoolchildren were analysed. After stool examination, 66 children were diagnosed to be infected with Giardia duodenalis, 22 with other protozoa, and 73 non-parasitized. The mean (+ 2 SD) values of secretory IgA in the non-parasitized group was considered as the criterion of positivity. The levels of total and specific anti-Giardia sIgA were significantly higher in children with Giardia compared with the group with other protozoa (p < 0.01) and the non-parasitized group (p < 0.001). The ELISA technique developed showed values of sensitivity and specificity of 74 and 94 per cent, respectively, a predictive value of 92 per cent for positive samples and 80 per cent for negative samples. Specific anti-Giardia IgA serum levels showed a low sensitivity (57 per cent) and a predictive value for negative samples (53 per cent). Our results suggest that secretory anti-Giardia IgA levels measured in saliva samples may reflect local intestinal IgA responses elicited by these parasites. Thus, determinations of the levels of sIgA anti-Giardia could be a useful diagnostic tool for giardiasis in children.

The association reactions of benzene molecules with alkali ions M(+) (Li(+), Na(+) and K(+)) under single collision conditions have been studied using a radiofrequency-guided-ion-beam apparatus and mass spectrometry characterization of the different adducts. Cross-section energy dependences for [M-C(6)H(6)](+) adduct formation have been measured at collision energies up to 1.20 eV in the center of mass frame. All excitation functions decrease when collision energy increases, showing the expected behaviour for barrierless reactions. From ab initio chemical structure calculations at the MP2(full) level, the formation of the adducts makes evident the alkali ion-benzene non-covalent chemical bond. The cross-section energy dependence and the role of radiative cooling rates and unimolecular decomposition on the stabilization of the energized collision complex are also discussed.

The association reactions between Li(+), K(+), and Rb(+) (M) and butanone and cyclohexanone molecules under single collision conditions have been studied using a radiofrequency-guided ion-beam apparatus, characterizing the adducts by mass spectrometry. The excitation function for the [M-(molecule)](+) adducts (in arbitrary units) has been obtained at low collision energies in the 0.10 eV up to a few eV range in the center of mass frame. The measured relative cross sections decrease when collision energy increases, showing the expected energy dependence for adduct formation. The energetics and structure of the different adducts have been calculated ab initio at the MP2(full) level, showing that the M(+)-molecule interaction takes place through the carbonyl oxygen atom, as an example of a nontypical covalent chemical bond. The cross-section energy dependence and the role of radiative cooling rates allowing the stabilization of the collision complexes are also discussed.

Collisions between potassium ions and neutral i-C3H7Br and i-C3H7OH, all in their electronic ground state, have been studied in the 0.10-10.00 eV center of mass (CM) collision energy range, using the radiofrequency-guided ion beam technique. In K(+) + i-C3H7Br collisions KHBr(+) formation was observed and quantified, while the analogous KH2O(+) formation in K(+) + i-C3H7OH was hardly detected. Moreover, formation of the ion-molecule adducts and their decomposition leading to C3H7(+) and either KBr or KOH, respectively, have been observed. For all these processes, absolute cross-sections were measured as a function of the CM collision energy. Ab initio structure calculations at the MP2 level have given information about the potential energy surfaces (PESs) involved. In these, different stationary points have been characterized using the reaction coordinate method, their connectivity being ensured by using the intrinsic-reaction-coordinate method. From the measured excitation function for KHBr(+) formation the corresponding thermal rate constant at 303 K has been calculated. The topology of the calculated PESs allows an interpretation of the main features of the reaction dynamics of both systems, and in particular evidence the important role played by the potential energy wells in controlling the reactivity for the different reaction channels.

Dehydrohalogenation and dehydration reactions of gas-phase i-C3H7Br and i-C3H7OH molecules induced by collision with Na(+), all participants being in their electronic ground state, were studied experimentally in our laboratory using a radiofrequency-guided ion beam apparatus and covering the 0.10-10.00 eV center of mass (CM) energy range. In Na(+) + i-C3H7Br collisions the formation of [C3H6-Na](+) and [HBr-Na](+) by dehydrohalogenation was observed and quantified, as well as that of the ion-molecule adduct [Na-i-C3H7Br](+) together with its decomposition products C3H7(+) and NaBr. In Na(+) + i-C3H7OH collisions the dehydration product [H2O-Na](+) was also found, while [C3H6-Na](+) was hardly detected. Moreover, the [Na-i-C3H7OH](+) adduct formation as well as its decomposition into C3H7(+) and NaOH were also quantified. For all these processes, absolute reaction cross sections were measured as a function of the CM collision energy. From measured excitation functions, rate constants for the formation of [C3H6-Na](+), [HBr-Na](+), and [H2O-Na](+) at 303 K were obtained. Complementing the experiments, exhaustive ab initio structure calculations at the MP2 level of theory were performed, giving information on the most relevant features of the potential energy surfaces (PESs) where the dehydrohalogenation, dehydration, and decomposition reactions take place adiabatically for both collision systems. On these PESs different stationary points associated with potential energy minima and transition state barriers were characterized, and their connectivity was ensured using the intrinsic-reaction-coordinate method. The main topology features of the ab initio calculated PESs allowed a qualitative interpretation of the experimental data also exposing the role of the sodium ion as a catalyst in elimination reactions.

Background: We evaluated the influence of intestinal parasitic infection on food sensitization associated to the severity of Atopic Dermatitis (AD) in a group of Warao Amerindian pre-school children. Methods: Feces examinations were performed in fresh stool specimens. Diagnosis of AD was done according to Hannifin and Rajka criteria and SCORAD index. Skin prick tests (SPT) were performed using extracts of cow's milk (CM), hen's egg (HE), Dermatophagoides pteronyssinus and Dermatophagoides farinae . Serum CM and HE-IgE levels (ELISA) were measured. Quantikine (R&D systems) assays were used for the determination of IL-13, TNF-a IL-6, and sCD23 in supernatants of CM-and HE-whole blood stimulated samples. Results: Atopic Dermatitis was reported in 23% of the children. It was significantly (p < 0.0001) associated toward both CM and HE-SPT positivity (p < 0.001). Giardia duodenalis infection (37%) was associated to the presence of AD (p = 0.005) and to a significant increase in the levels of CM stimulated TNF-a (p ¼ 0:006), IL-13 (p = 0.01), sCD23 (p = 0.001), CM-IgE (p = 0.012) and CM-SPT (p < 0.0001). Similarly, G. duodenalis infection was particularly associated with the increase on the levels of HE-stimulated TNF-a (p = 0.001), sCD23 (p = 0.001), HE-IgE levels (p = 0.002) and HE-SPT (p = 0.001). Conclusion: Gut inflammation caused by G. duodenalis may enhance food allergic reactivity contributing to the manifestation of AD in these children. However, other environmental factors (not considered in this work) as well as an atopic background among the Warao population would also contribute to the presence of AD.

THE role of the L-arginine-nitric oxide metabolic pathway was explored for interleukin-2-induced proliferation in the cytotoxic T lymphocyte clone CTLL-2. Specific inhibition of nitric oxide synthase significantly diminished, in a concentration-dependent manner, 3H-thymidine uptake of CTLL-2 cells in response to different concentrations of interleukin 2. Withdrawal of L-arginine from culture medium resulted as potent as the higher inhibition obtained when blocking nitric oxide synthase with L-arginine analogues. Furthermore, intermedial concentrations of Larginine and exogenous nitric oxide donors were found for achieving optimal IL2-induced proliferation of CTLL-2. These findings prompted us to suggest that intra-and/or inter-cellular nitric oxide signalling may contribute to the modulation of the IL2 mitogenic effect upon cytotoxic T lymphocytes.

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