Ochratoxin a determination in beer by solid-phase microextraction coupled to liquid chromatography with fluorescence detection: a fast and sensitive method for assessment of noncompliance to legal limits

A solid-phase microextraction-liquid chromatography-fluorescence detection (SPME-LC-FD) method for the determination of ochratoxin A (OTA) in commercial beer samples was developed for the first time using a 60 microm thick poly(dimethylsiloxane)/divinylbenzene (PDMS/DVB) fiber. The procedure required a very simple sample pretreatment, an isocratic elution, and provides a selective extraction. All of the factors influencing fiber adsorption (extraction time, temperature, pH, and salt addition) and desorption of the analyte (desorption and injection time and desorption solvent mixture composition) have been investigated. The linear range investigated in beer was 0.03-2 ng/mL; within-day and between-days relative standard deviation in beer were 4.3 and 5.9%, respectively. The limit of quantification in spiked beer was 53 pg mL(-)(1), well below all European regulatory levels.     

Carotenoid profiling and biosynthetic gene expression in flesh and peel of wild-type and hp-1 tomato fruit under UV-B depletion

Although light is recognized as one of the main factors influencing fruit carotenogenesis, the specific role of UV-B radiation has been poorly investigated. The present work is addressed to assess the molecular events underlying carotenoid accumulation in presence or absence of ultraviolet-B (UV-B) light in tomato fruits of wild-type and high pigment-1 (hp-1), a mutant characterized by exaggerated photoresponsiveness and increased fruit pigmentation. Gene expression analyses indicated that in wild-type fruits UV-B radiation mainly negatively affects the carotenoid biosynthetic genes encoding enzymes downstream of lycopene both in flesh and peel, suggesting that the down-regulation of genes CrtL-b and CrtL-e and the subsequent accumulation of lycopene during tomato ripening are determined at least in part by UV-B light. In contrast to wild-type, UV-B depletion did not greatly affect carotenoid accumulation in hp-1 and generally determined minor differences in gene expression between control and UV-B-depleted conditions.     

Effects of blood orange juice intake on antioxidant bioavailability and on different markers related to oxidative stress

Orange juice is a source of antioxidants that might afford in vivo protection from oxidative stress. To test this hypothesis, we carried out a human intervention study with blood orange juice containing high amounts of vitamin C, anthocyanins, and carotenoids. Sixteen healthy female volunteers were enrolled in a crossover study and were given 600 mL/day of blood orange juice or a diet without juice for 21 days. Before and after each intervention period, plasma vitamin C, cyanidin-3-glucoside, and carotenoids were quantified. Furthermore, plasma antioxidant capacity, malondialdehyde concentration in plasma, 11-dehydrotromboxane B(2) urinary excretion, and lymphocyte DNA damage were evaluated as biomarkers of oxidative stress. Blood orange juice consumption determined a significant increase in plasma vitamin C, cyanidin-3-glucoside, beta-cryptoxanthin, and zeaxanthin. Also, lymphocyte DNA resistance to oxidative stress was improved whereas no effect was observed on the other markers that we analyzed. In turn, these results suggest that blood orange juice is a bioavailable source of antioxidants, which might moderately improve the antioxidant defense system; however, the long-term effects of its consumption are to be further investigated.     

Tertiary hypothyroidism in a dog

: A nine-year-old male entire Labrador was diagnosed with pituitary dependent hyperadrenocorticism. Following seven months of successful mitotane therapy, the dog presented with marked weight gain, seborrhoea and alopecia. Routine clinicopathological analyses revealed marked hypercholesterolaemia. Serum total and free thyroxine (T4) concentrations were below their respective reference ranges. Serum thyroid stimulating hormone (cTSH) concentration was within reference range. TSH and thyrotropin releasing hormone (TRH) response tests revealed adequate stimulation of total T4 in both, and cTSH in the latter test. Magnetic resonance imaging revealed a mass arising from the pituitary fossa, with suprasellar extension. A diagnosis of tertiary hypothyroidism was made. Following four weeks of levothyroxine therapy, circulating cholesterol concentration had declined, weight loss had ensued and dermatological abnormalities had improved. Euthanasia was performed four months later due to the development of neurological signs. A highly infiltrative pituitary adenoma, with effacement of the overlying hypothalamus was identified on post mortem examination. Tertiary hypothyroidism has not been previously reported in dogs.     

Use of Red Blood Cell Membranes to Evaluate the Antioxidant Potential of Plant Extracts

Antioxidant phytochemicals in fruits and vegetables of a vegetarian diet may account for the reduced risk of aging and stress oxidative associated diseases. In this study, a simple, rapid and accurate new bioassay for the determination of the antioxidant activity of purified or crude plant extracts and thier interactions is described, based on the fluorimetric determination of thiobarbituric acid reactive substances (TBARS) released by UV-B radiated red blood cell (RBC) ghosts. Pure resveratrol, white and red wine and pomegranate juice (PJ) were used as antioxidant source to test the biological method. TBARS production is a function of radiation time, the number of RBC ghosts in the radiated sample and the loaded antioxidant. The antioxidant activity of resveratrol was detected at a submicromolar concentration range [0.02 μg/mL–0.1 μmol/L]. The activity of red wine was almost 10 times higher than that of white wine, and PJ juice had the highest activity. Submaximal protective effects of PJ and red wine were additive.     

Phenolic compounds and saponins in quinoa samples (Chenopodium quinoa Willd.) grown under different saline and nonsaline irrigation regimens

Quinoa is a pseudocereal from South America that has received increased interest around the world because it is a good source of different nutrients and rich in antioxidant compounds. Thus, this study has focused on the effects of different agronomic variables, such as irrigation and salinity, on the phenolic and saponin profiles of quinoa. It was observed that irrigation with 25% of full water restitution, with and without the addition of salt, was associated with increases in free phenolic compounds of 23.16 and 26.27%, respectively. In contrast, bound phenolic compounds were not affected by environmental stresses. Saponins decreased if samples were exposed to drought and saline regimens. In situations of severe water deficit, the saponins content decreased 45%, and 50% when a salt stress was added. The results suggest that irrigation and salinity may regulate the production of bioactive compounds in quinoa, influencing its nutritional and industrial values.     

<Emphasis Type="Italic">Sesuvium portulacastrum</Emphasis> (L.) L. a promising halophyte: cultivation,utilization and distribution in India

Seapurslane, Sesuvium portulacastrum (L.) L. (Aizoaceae) is a pioneer, psammophytic facultative halophyte naturally growing in the subtropical, mediterranean, coastal and warmer zones of the world. The plant is utilized as a vegetable by the local peoples as well as forage for domestic animals in the coastal area. Environmentally, it is utilized for the bioreclamation of saline soil in the arid and semiarid regions and an alternative culture to problematic soils. The Sesuvium clones collected from the coastal regions of India lead to conclusions with respect to its geographical distribution and prospective utilization in the bioremediation of coastal and saline areas, as an emerging source of secondary metabolite-phytoecdysteroids in sericulture industry and cultivation on large scale for food as well as forage, of this underutilized, neglected wild and also cultivated plant species.     

Scleral rupture in dogs, cats, and horses

OBJECTIVES: The aim of this retrospective study was to summarize the most frequent clinical signs, ultrasonographic, and histological findings accompanying scleral rupture as a result of blunt trauma in dogs, cats, and horses. ANIMALS STUDIED AND PROCEDURES: Thirty small animals and three horses diagnosed with scleral rupture resulting from blunt trauma. B-mode ultrasonography was performed on 20 animals. Histopathology was carried out on 18 enucleated globes. RESULTS: In small animals, 80% presented hyphema, 60% subconjunctival hemorrhage, and 53% eyelid and conjunctival swelling. In horses, 100% presented eyelid and conjunctival swelling, 67% hyphema, subconjunctival hemorrhage, and collapsed anterior chamber. Ultrasonographic findings were an area with ill-defined scleral margins (90%), echoic/hyperechoic contents in the anterior and posterior chamber (55%) and in the vitreous (80%). In small animals, scleral rupture location noted on gross examination was: at the posterior pole (4), close to the optic nerve (3), near the limbus (2), and in the dorsal aspect of the globe (1). In horses, the lesion was located at the limbus (3). In small animals, histopathology showed presence of hemorrhage in the anterior, posterior chamber, and vitreous (94%), retinal detachment (94%), choroidal edema and hemorrhages (88%), and choroidal detachment as a result of suprachoroidal hemorrhage (88%). The same lesions were found in the globes of two horses. In small animals, rupture location noted on histopathology was: at the posterior pole (8), close to the optic nerve (4), near the limbus (1), near the ciliary body (1). CONCLUSIONS: The most frequent clinical signs observed were hyphema, subconjunctival hemorrhage, and eyelid and conjunctival swelling. Ultrasonographic findings suggestive for scleral rupture were ill-defined scleral borders and/or echoic/hyperechoic material in the cavities of the globe. On histopathology, lesions severely altering the anatomy of the eye structures were: hemorrhage into the chambers of the globe, subretinal and suprachoroidal hemorrhage leading to retinal and choroidal detachment, respectively. In small animals, the most frequent locations for scleral rupture were the posterior pole and close to the optic nerve, whereas in horses it was the limbus.