Influence of Marolo (Annona crassiflora Mart.) Pulp Intake on the Modulation of Mutagenic/Antimutagenic Processes and Its Action on Oxidative Stress In Vivo

Brazilian savanna constitutes a valuable ecoregion that contributes to the supply of fruit known worldwide for its nutritional value and peculiar flavors. Among them, the Marolo (Annona crassiflora Mart.) fruit is known for its use in folk medicine. In order to establish the safety of Marolo for human consumption, this study evaluated the following: the chemical composition of Marolo pulp; its mutagenic and antimutagenic activities using micronucleus test; and the oxidative stress induced in the livers of mice fed a diet containing 1%, 10% or 20% pulp. It was observed that the chemical composition of marolo pulp was similar to that of common fruit; nevertheless, its lipidic content and energetic values were higher. In the mice fed experimental diets, the biochemical parameters of the blood serum showed normal levels of glucose, triglycerides, and cholesterol. The micronucleus test indicated neither mutagenic nor antimutagenic effects of Marolo consumption on bone marrow cells but showed potentialization of cyclophosphamide (CP). The oxidative stress levels observed indicated that CP was not exerting a great influence on the induction of reactive oxygen species. As the whole fruit is a complex matrix, the interactions between its components could be responsible for its negative and positive biological effects.     

A quantitative stable-isotope LC-MS method for the determination of folic acid in fortified foods

A stable-isotope liquid chromatography-mass spectrometry (LC-MS) assay was developed for the quantitative determination of folic acid in fortified foods. Folic acid was extracted from food samples into a phosphate buffer, purified on a C-18 Sep-Pak cartridge, and analyzed by LC-MS in the negative ion mode using electrospray ionization. The analyte was quantified using (13)C(5)-folic acid as an internal standard. The coefficient of variation for the precision of the method was 5.6% based on the analysis of four sample replicates. The accuracy of the method was assessed using a standard method of addition of folic acid to a shredded whole-wheat cereal. The quantitative determination of folic acid in this matrix was linear over 1 order of magnitude having a concentration range of 2.4 to 24 microg/g of food (or 0.05 to 0.5 microg of analyte injected into the LC-MS). The overall quantitative efficiency of the method was evaluated using a standard reference material (infant formula SRM 1846). The method was applied to the determination of folic acid in several test samples (fortified breakfast cereals), and the values were in accord with the manufacturer's claim. This method advances a LC-MS technique for the determination of folic acid in fortified foods based on stable-isotope dilution methodology. The specificity of the technique and quantitative accuracy of the method in various food substrates suggests that the method may be adapted for routine analysis in other fortified foods.     

Endotracheal nebulization of N-acetylcysteine increases airway resistance in cats with experimental asthma

N-acetylcysteine (NAC), a mucolytic and antioxidant, is speculated to cause bronchoconstriction in cats when delivered via aerosol. We hypothesized that in cats with experimental asthma, aerosol delivery of NAC (400mg cumulative dose) via an endotracheal tube would increase airflow limitation as measured by ventilator-acquired mechanics. After endotracheal drug delivery, airway resistance and inspiratory plateau pressure (Pplat) measurements were obtained in six mechanically ventilated asthmatic cats. Results demonstrated significantly increased airway resistance (P=0.0007) compared with aerosolized saline control; Pplats were not significantly different (P=0.059). All cats exhibited at least one adverse effect: excessive airway secretions (n=3), spontaneous cough (n=2), unilateral strabismus (n=1) and post-anesthetic death (n=1). No adverse reactions were noted with saline aerosol; cough was noted in one cat with methacholine challenge. In conclusion, airway resistance and adverse reactions were documented in all cats after NAC aerosol delivery. Further studies must be performed to evaluate if it is an effective mucolytic and/or antioxidant in cats and to determine if bronchodilator pre-treatment will negate NAC-induced bronchoconstriction.     

Molecular cloning and tissue-specific expression of Toll-like receptor 5 gene from turkeys

Toll-like receptors (TLRs), a family of transmembrane and cytosolic proteins, detect microbial patterns, initiating innate immune responses in various organisms. Although they are abundant, genetic characterization and functional differences of TLRs in economically important avian species such as chickens and turkeys have not been investigated in detail. In this study, the putative TLR5 coding region from turkey genome was sequenced, and its homology to other vertebrate species was analyzed. Secondary structure analysis revealed protein motifs typical of the chicken TLR5 protein structure, with 97% amino acid identity between them. mRNA expression profiling in adult turkeys revealed abundant TLR5 expression in a broad range of tissues. Stimulation with the TLR5 ligand flagellin resulted in the production of the inflammatory mediators interleukin (IL)-1beta, IL-6, and nitric oxide in peripheral blood mononuclear cells. To our knowledge, this is the first complete turkey TLR5 coding DNA sequence reported in sequence databases.     

Pyometra in a 13-year-old neutered queen

A 13-year-old female domestic shorthair cat was examined because of weight loss over 4 months. The history included being neutered when a kitten and no signs of estrus recognized by the owner since this surgery. A mid-abdominal mass was palpated. Ultrasonographic abdominal evaluation revealed a tubular fluid-filled mass. The cat underwent surgery to remove the mass, which was one horn of the uterus. The patient made a full recovery.     

Effect of Fasciola gigantica infection on adrenal and thyroid glands of riverine buffaloes

Effect of Fasciola gigantica infection on adrenal and thyroid glands was investigated using eight male, yearling Murrah buffaloes. The animals were randomly assigned to two groups of four buffaloes each (Group-A, infected; Group-B, non-infected control). Animals of Group-A were orally infected with 1000 F. gigantica viable metacercariae, keeping other four animals of Group-B as uninfected control. In the infected buffaloes, the clinical signs began appearing from 7th week postinfection (p.i.) and eggs were detected in the faeces between day 93 and 99 (95.5+/-1.25) postinfection (p.i.). The serum cortisol level, revealed a significant (P<0.05) rise during initial stage of the infection, followed by a continuous fall from 12th week onward. Peak cortisol level on 10th week (13.30+/-2.57ngml(-1)) was associated with eosinophilia (11.0+/-0.95%). However, non-infected controls maintained almost uniform cortisol levels (3.97+/-0.15-5.88+/-0.09ngml(-1)) throughout the period of the study. The pathological changes of adrenal glands were correlated with physiological dysfunction of the glands. The levels of T(3) and T(4) were significantly (P<0.05-0.01) low from 14th week onward and were synchronous with in situ migration, growth and development of F. gigantica. Significant reduction in the thyroid hormones was further supported by histopathological evidence of lymphocytic thyroiditis confirming hypothyroidism. A decrease in Hb, PCV, total erythrocyte counts and appearance of reticulocytes in the blood of the infected buffaloes suggested regenerative anemia, which could partly be due to hypothyroidism.     

A potential krimpsiekte vaccine

Krimpsiekte, a chronic form of cardiac glycoside poisoning, is an important plant-induced intoxication of small stock in South Africa. It is caused by cumulative, neurotoxic bufadienolides, such as cotyledoside. A cotyledoside-bovine serum albumin conjugate was synthesized to immunize animals. The efficacy of the cotyledoside-conjugate in inducing an immunological response was ascertained in rabbits (n = 4) and sheep (n = 4) by determining cotyledoside antibody titres with an ELISA using cotyledoside-hen ovalbumin as antigen. The formation of anticotyledoside antibodies was induced in both rabbits and sheep following immunization with the cotyledoside-protein conjugate. Protection provided by the vaccine was demonstrated by challenging sheep (n = 4) with repeated, daily doses of cotyledoside (0.015 mg/kg) administered intravenously, commencing 45 days after the initial vaccination. One control animal died on Day 3 of the challenge period and the other was severely affected after administration of the third cotyledoside dose. The immunized ewes (n = 2) remained clinically unaffected and the challenge was suspended following six daily injections. Vaccination as a means of preventing krimpsiekte seems to be quite feasible and deserves further investigation.     

Cosorption of atrazine and a lauryl polyoxyethylene oxide nonionic surfactant on smectite

Commercial atrazine formulations commonly contain nonionic surfactants that serve as solubilizing and wetting agents for enhancing the stability and efficacy of the herbicide. The fate of atrazine in soils has been extensively investigated; yet, few studies have considered the effects of formulation components on the fate of atrazine in soils. In this study, we investigated the influence of the nonionic surfactant, Brij 35 (Brij), on the sorption of atrazine on Ca- and K-saturated samples of a reference smectite, Panther Creek (PC). In general, Brij concentrations of 50 and 200 mg L(-1) had little effect on atrazine sorption, but sorption was substantially inhibited by Brij concentrations of 2100 mg L(-1). For Brij concentrations of 6300 mg L(-1), atrazine sorption was intermediate between that observed for the 200 and 2100 mg L(-1) Brij systems. Brij molecules themselves were very strongly sorbed by PC, with sorption maxima exceeding 200 g kg(-1). X-ray diffraction analysis of Brij-treated PC indicated that the sorbed Brij was intercalated into interlayers of the smectite. At Brij concentrations of 2100 mg L(-1), Brij competed with atrazine for interlayer sorption sites. In contrast, at the initial Brij concentration of 6300 mg L(-1), the clay interlayers were largely filled with Brij, and excess Brij probably accumulated on external surfaces of the clay as surface micelles. We hypothesize that atrazine partitioning into surfactant micelles on external surfaces of the clay led to enhanced retention by the solid phase.