Clinical evaluation of pimecrolimus eye drops for treatment of canine keratoconjunctivitis sicca: a comparison with cyclosporine A

The aim of this study was to evaluate the efficacy of pimecrolimus oil-based eye drops in alleviating the clinical signs of keratoconjunctivitis sicca (KCS) in dogs and to compare the efficacy with that of cyclosporine A (CsA) ointment. An open-label, multicenter study enrolling 44 dogs previously untreated with CsA was conducted. Dogs were randomly assigned to a treatment group and medicated twice daily for 8 weeks. After that time the mean increase (+/-SEM) in the Schirmer tear test was 9.2+/-1.6 mm/min in the pimecrolimus group and 5.8+/-1.1 mm/min in the CsA group (P=0.085). The improvement in clinical signs of inflammation in eyes treated with pimecrolimus was significantly greater than in eyes treated with CsA (P=0.02). The results show that 1% pimecrolimus oily eye drops are as safe as and more effective than CsA ointment in controlling KCS in dogs.     

Effect of red wine marinades on the formation of heterocyclic amines in fried chicken breast

Genotoxic heterocyclic amines (HAs) are formed via the Maillard reaction and free radical reaction mechanisms when meat or fish is cooked at usual cooking conditions. In this paper, the effect of the addition of red wine was tested to study if it interferes in HA formation. Fried chicken breast was the food item chosen, and three different red wines, characterized in terms of grape varieties, free amino acids, antioxidant properties, and metallic composition, were used to marinate meat prior to the heating process. Unmarinated samples and samples marinated with an ethanol-water mixture provided reference HA levels. The frying experiments were performed under well-controlled temperature and time conditions. The samples were analyzed for HA content using solid-phase extraction and LC-MS/MS. DMIP, PhIP, MeIQx, 4,8-DiMeIQx, IFP, TMIP, harman, and norharman were identified in fried chicken breast. Red wine marinades were found to reduce the formation of some of the HAs formed. PhIP, with a reduction of up to 88%, was the most minimized amine, although the formation of harman was enhanced.     

Rapid determination of nonaromatic organic acids in honey by capillary zone electrophoresis with direct ultraviolet detection

A rapid capillary zone electrophoresis (CZE) method with direct ultraviolet (UV) detection has been set up and developed to determine the most important nonaromatic organic acids in honey with a really simple treatment of the sample. The determination of oxalic, formic, malic, succinic, pyruvic, acetic, lactic, citric, and gluconic acids has been carried out in 4 min. The electrolyte composition was phosphate as the carrier buffer (7.5 mM NaH(2)PO(4) and 2.5 mM Na(2)HPO(4)), 2.5 mM tetradecyltrimethylammonium hydroxide (TTAOH) as electroosmotic flow modifier, and 0.24 mM CaCl(2) as selectivity modifier, with the pH adjusted at 6.40 constant value. The running voltage was -25 kV at a thermostated temperature of 25 degrees C. The injections were performed in hydrodynamic mode (30 s), and the detection mode was UV direct at 185 nm. Validation parameters of the method as detection and quantification limits, linearity, precision (repeatability and reproducibility), and recovery were also studied. The advantages related to the technique such as simplicity, short analysis times, and low consumption of chemicals as well as the good validation parameters obtained for this method permit it to be considered as adequate for routine analysis in honey.     

Urolithins, ellagic acid-derived metabolites produced by human colonic microflora, exhibit estrogenic and antiestrogenic activities

Urolithins A and B (hydroxy-6H-dibenzo[b,d]pyran-6-one derivatives) are colonic microflora metabolites recently proposed as biomarkers of human exposure to dietary ellagic acid derivatives. Molecular models suggest that urolithins could display estrogenic and/or antiestrogenic activity. To this purpose, both urolithins and other known phytoestrogens (genistein, daidzein, resveratrol, and enterolactone) were assayed to evaluate the capacity to induce cell proliferation on the estrogen-sensitive human breast cancer MCF-7 cells as well as the ability to bind to alpha- and beta-estrogen receptors. Both urolithins A and B showed estrogenic activity in a dose-dependent manner even at high concentrations (40 microM), without antiproliferative or toxic effects, whereas the other phytoestrogens inhibited cell proliferation at high concentrations. Overall, urolithins showed weaker estrogenic activity than the other phytoestrogens. However, both urolithins displayed slightly higher antiestrogenic activity (antagonized the growth promotion effect of 17-beta-estradiol in a dose-dependent manner) than the other phytoestrogens. The IC(50) values for the ERalpha and ERbeta binding assays were 0.4 and 0.75 microM for urolithin A; 20 and 11 microM for urolithin B; 3 and 0.02 for genistein; and 2.3 and 1 for daidzein, respectively; no binding was detected for resveratrol and enterolactone. Urolithins A and B entered into MCF-7 cells and were metabolized to yield mainly urolithin-sulfate derivatives. These results, together with previous studies regarding absorption and metabolism of dietary ellagitannins and ellagic acid in humans, suggest that the gut microflora metabolites urolithins are potential endocrine-disrupting molecules, which could resemble other described "enterophytoestrogens" (microflora-derived metabolites with estrogenic/antiestrogenic activity). Further research is warranted to evaluate the possible role of ellagitannins and ellagic acid as dietary "pro-phytoestrogens".     

Squash-prep cytology in the diagnosis of canine and feline nervous system lesions: a study of 42 cases

BACKGROUND: The increased sophistication of imaging techniques in veterinary medicine allows the detection of a wide variety of intracranial and intraspinal lesions; however, imaging often does not provide a definitive diagnosis for nervous system (NS) lesions. Cytology is emerging as a useful diagnostic tool for obtaining a fast and accurate assessment of NS lesions, but little information is available for dogs and cats. OBJECTIVES: The purpose of this study was to assess the accuracy of cytologic evaluation of squash samples from NS lesions in dogs and cats and to consider cytology-based diagnostic guidelines and sources of misdiagnosis. METHODS: Cytologic specimens from masses localized in the central and peripheral NS taken during surgery or postmortem examination were classified into 3 groups according to the final histopathologic diagnosis: Group 1 = completely correct diagnosis, when the cytologic diagnosis and final histologic diagnosis were exactly correlated; Group 2 = partial correlation, when the cytologic diagnosis only partially correlated with the final histologic diagnosis, and Group 3 = no correlation, when the cytologic diagnosis was incorrect and there was no correlation with the general histologic type of lesion. The diagnostic accuracy of cytopathology was calculated by considering the histopathologic diagnosis as the "gold standard," and calculating a 95% confidence interval (CI). RESULTS: A total of 42 animals (33 dogs and 9 cats) were included in the study. The cytologic diagnoses were classified in Group 1 for 32 cases (76%; 95% CI 0.63-0.89), in Group 2 for 6 cases (14%; 95% CI 0.04-0.25), and in Group 3 for 4 cases (10%; 95% CI 0.006-0.18). Considering both complete and partial correlation as an adequate result, cytologic diagnosis was satisfactory in 90% of biopsies. CONCLUSIONS: Although the current series of cases is relatively small, cytologic evaluation of squash preparations can be considered a fairly accurate and reliable tool in the diagnosis of NS lesions.     

Phlorotannin Extracts from Fucales Characterized by HPLC-DAD-ESI-MSn: Approaches to Hyaluronidase Inhibitory Capacity and Antioxidant Properties

Purified phlorotannin extracts from four brown seaweeds (Cystoseira nodicaulis (Withering) M. Roberts, Cystoseira tamariscifolia (Hudson) Papenfuss, Cystoseira usneoides (Linnaeus) M. Roberts and Fucus spiralis Linnaeus), were characterized by HPLC-DAD-ESI-MSⁿ. Fucophloroethol, fucodiphloroethol, fucotriphloroethol, 7-phloroeckol, phlorofucofuroeckol and bieckol/dieckol were identified. The antioxidant activity and the hyaluronidase (HAase) inhibitory capacity exhibited by the extracts were also assessed. A correlation between the extracts activity and their chemical composition was established. F. spiralis, the species presenting higher molecular weight phlorotannins, generally displayed the strongest lipid peroxidation inhibitory activity (IC₅₀ = 2.32 mg/mL dry weight) and the strongest HAase inhibitory capacity (IC₅₀ = 0.73 mg/mL dry weight). As for superoxide radical scavenging, C. nodicaulis was the most efficient species (IC₅₀ = 0.93 mg/mL dry weight), followed by F. spiralis (IC₅₀ = 1.30 mg/mL dry weight). These results show that purified phlorotannin extracts have potent capabilities for preventing and slowing down the skin aging process, which is mainly associated with free radical damage and with the reduction of hyaluronic acid concentration, characteristic of the process.     

Fecundity of the sea louse Caligus rogercresseyi on its native host Eleginops maclovinus captured near salmon farms in southern Chile

The sea louse (Caligus rogercresseyi) is the most significant parasitic pathogen in Chilean salmon farms, and it infects farmed salmon and native host fish. Fecundity is one of the most important parameters for understanding the population dynamics of a species; however, this information is scarce for this parasite. The fecundity of C. rogercresseyi females collected from native hosts (Eleginops maclovinus) captured near salmon farms in southern Chile was measured to evaluate the reproductive output of this parasite on this host fish. From June 2008 to May 2009, 212 specimens of E. maclovinus were examined. Each fish was measured, and all its parasites were collected, sorted and counted. Seventy‐nine ovigerous C. rogercresseyi females (OFs) were measured. Total body length, egg string length and total number of eggs per string were recorded for each parasite. Ovigerous females body length varied between 3.9 and 5.0 mm. Fecundity varied between 12 and 56 eggs string^?1, and it was correlated with OF body length. Temporal variations in OF fecundity were explained by co‐variation in OF body length, but not by month. Ovigerous females on E. maclovinus were smaller and showed lower fecundity than OFs on farmed salmon. Our results suggest that native hosts play a secondary role in C. rogercresseyi egg production in Codigue bay.     

Hydrogen peroxide,iodine solution and methylene solution highly enhance the hatching rate of freshwater ornamental fish species

The aim of this study was to evaluate the effect of hydrogen peroxide, iodine solution (PVP) and methylene blue on eggs disinfection of three ornamental fish species, Danio rerio, Pterophyllum scalare and Gymnocorymbus ternetzi. The main idea was to create conditions to enhance the hatching rates. Eggs of each species were exposed to different concentrations of hydrogen peroxide (5, 10, 15 and 25 mg/L), PVP (0.25, 0.5, 0.75 and 1 mg/L) and methylene blue (0.5 1, 2 and 3 mg/L). The optimal doses ranged between species and chemicals: for G. ternetzi, the concentrations that high enhanced the hatching rate were 1 mg/L for the PVP treatment, 25 mg/L for the hydrogen peroxide treatment and 3 mg/L for methylene blue treatment; for P. scalare, the best results were achieved with 25 mg/L for hydrogen peroxide treatment and 3 mg/L for methylene blue treatment. By contrast, for all the different chemical did not increased the D. rerio hatching rate. Results showed that hydrogen peroxide and methylene blue are the most versatile, effective and safe to use in these species. On the other hand, PVP can be used but with many precautions due to very low safety margin. Results clearly show that the optimal concentration of chemicals for eggs disinfection is fish species dependent and it is completely wrong to extrapolate concentrations between different chemicals and fish species. Our study suggests that P. scalare can be used as a model in study of effectiveness of new chemicals with potential to disinfect water and increase hatching rates.     

Acclimation of cuttings from different willow genotypes to flooding depth level

Climate change will increase the frequency of extreme rain events, causing more flooding episodes. Willows are usually planted in marginal lands like flood prone areas. For willow plantations developed from rootless cuttings, the establishment phase is crucial, because the cuttings are still developing a shoot and root system and have a higher vulnerability to stress. A flooding episode during this early period may have a negative effect upon plants. We analyzed the responses to flooding of eight willow genotypes, representing important species from the economic and ecological point of view (Salix alba, S. matsudana, S. amygdaloides and S. matsudana × S. nigra hybrids). The treatments started when the plants were 2 months old and lasted for 3 weeks. They were identified as: Control (watered to field capacity); F10 (plants submerged 10 cm above soil surface) and F50 (plants submerged 50 cm above soil surface). The F50 treatment showed a greater growth reduction than the F10 treatment in most clones, either measured in height, diameter or total biomass. Both flooding treatments reduced significantly the root-to-shoot ratio compared to control plants. The F50 treatment increased the leaf nitrogen content and specific leaf area in all genotypes. Both treatments changed plant and leaf traits in different ways according to the depth of the floodwater. These changes may have lasting effects on growth recovery in the post-flooding period. Willow clones with a fast initial growth will be the best option to minimize growth reduction in areas prone to experience flooding episodes.