Using the method of triads in the validation of a food frequency questionnaire to assess the consumption of fatty acids in adults.

BACKGROUND: It is of great value to develop valid instruments to estimate food consumption; for this purpose, the triads method has been applied in validation studies of dietary intake to evaluate the correlation between three measurements [food frequency questionnaire (FFQ), reference method and biomarker]. The main aim of the present study was to validate a FFQ for Brazilian adults by means of the method of triads by estimating the ingestion of total fatty acids based on the level of saturation. METHODS: The present study enrolled 152 Brazilian adults of both sexes, who were residents in the city of Viçosa, Brazil. The ingestion of total saturated, monounsaturated, polyunsaturated, trans, linoleic and linolenic fatty acids was assessed by means of a FFQ, two food records, and biomarkers, which were detected by gas chromatography. The validation coefficients were calculated using the method of triads and concordance was determined by Kappa statistics. RESULTS: The FFQ was considered an adequate dietary method, because, based on the validation coefficients, it estimates the intake of total fat (0.84), as well as linolenic (0.90) and linoleic acids (0.31). A high concordance rate was confirmed for all nutrients assessed by the FFQ and food records. Regarding the biomarkers, linolenic acid and linoleic acid presented greater concordance. CONCLUSIONS: According to the validation coefficients, the FFQ precisely estimated total fat, linolenic acid and linoleic acid contents.

Cloning and partial characterization of a Boophilus microplus (Acari: Ixodidae) glutathione S-transferase.

A cDNA of glutathione S-transferase (GST) was isolated from a cDNA library of salivary glands of Boophilus microplus. The recombinant protein was purified by glutathione affinity chromatography and assayed upon the chromogenic substrate CDNB. The 864 bp cloned fragment was sequenced and showed an open reading frame coding for a protein of 220 amino acids. Expression of the GST gene was tested by RT-PCR in tick tissues and larvae mRNA. Comparison of the deduced amino acid sequence with GSTs from other species revealed that the enzyme is closely related to the mammalian class mu GSTs.

Isolation of an aspartic proteinase precursor from the egg of a hard tick, Boophilus microplus.

An aspartic proteinase precursor, herein named BYC (Boophilus Yolk pro-Cathepsin) was isolated from eggs of the hard tick, Boophilus microplus. As judged by electrophoresis on sodium dodecyl sulfate polyacrylamide slab gel (SDS-PAGE), purified BYC presented 2 bands of 54 and 49 kDa, bearing the same NH2-terminal amino acid sequence. By Western blot analysis, BYC was also found in the haemolymph, indicating an extraovarian site of synthesis. Several organs were incubated in culture medium with [35S]methionine, and only the gut and fat body showed synthesis of BYC polypeptides. Protein sequencing of both the NH2-terminal and an internal sequence obtained after cyanogen bromide (CNBr) cleavage of BYC revealed homology with several aspartic proteinase precursors. Incubation at pH 3.5 resulted in autoproteolysis of BYC, which produced the mature form of the enzyme, that displayed pepstatin-sensitive hydrolytic activity against haemoglobin. Western blot analysis using anti-BYC monoclonal antibodies showed proteolytic processing of BYC during embryogenesis and suggested activation of the enzyme during development. A role of BYC in degradation of vitellin, the major yolk protein of tick eggs, is discussed.

Immunization of bovines with an aspartic proteinase precursor isolated from Boophilus microplus eggs.

The capacity of the Boophilus Yolk pro-Cathepsin (BYC) to induce a protective immune response in cattle against Boophilus microplus infestation was tested by vaccination experiments and by inoculation of monoclonal antibody (MAb) against BYC into fully engorged tick females. In immunization experiments the measurement of various biological parameters demonstrated a partial protection against B. microplus. A continuous decrease in the levels of specific antibodies was observed over 11 months when six bovines were maintained in field conditions. The inoculation of the MAb into tick females produced a dose-dependent decrease in oviposition and survival of the ectoparasite compared to the control.