Effect of 1-24ACTH administration on sheep blood granulocyte functions

The objective of the present study was to determine the efficiency of blood neutrophils (PMN) taken from sheep during acute stress. Ten healthy Charolle sheep were sampled before treatment (T0) and 1 (T1), 2 (T2), 24 (T24) and 48 (T48) hours after 1-24ACTH administration. Ten sheep serving as the controls were sampled at the same time intervals, using saline solution instead of 1-24ACTH. At each time sampling, rectal temperature, heart rate, cortisol, glucose, non-esterified fatty acids (NEFA), total and differential leukocyte counts were evaluated. PMN were isolated after centrifugation of whole blood and hypotonic lysis of RBC. Chemotaxis was evaluated on a modified Boyden chamber using a nitrate cellulose filter and both Zymosan activated serum (ZAS) and interleukin-8 (IL-8) as chemoattractants. Phagocytosis was measured using both non-opsonized latex beads and fluoresceinated yeasts opsonized with homologous serum. Superoxide (O(-)2) production was evaluated by measuring superoxide dismutase-inhibitable reduction of ferricytochrome C, and adherence by a colorimetric assay of acid phosphatase activity of adherent cells. The administration of 1-24ACTH induced an acute stress reaction, indicated by the presence of clinical, biochemical and hematological changes. Adherence significantly increased from T0 to T2 in treated sheep. This might be responsible for the depression of non-specific immunity in stressed animals. Studies using stressors other than 1-24 ACTH are needed to verify the influence of other components of the stress reaction on PMN functions.     

COMBINATION OF RADIATION THERAPY AND FIROCOXIB FOR THE TREATMENT OF CANINE NASAL CARCINOMA

Carcinomas represent two‐thirds of canine nasosinal neoplasms. Although radiation therapy (RT) is the standard of care, the incidence of local recurrence following treatment is high. Cyclooxygenase‐isoform‐2 (COX‐2) is expressed in 71–95% of canine nasal carcinomas and has been implicated in tumor growth and angiogenesis. Accordingly, COX‐2 inhibition seems rational to improve outcome. Dogs with histologically confirmed, previously untreated nasal carcinomas were randomized to receive the combination of a selective COX‐2 inhibitor (firocoxib) and palliative RT (Group 1) or RT and placebo (Group 2). Patients were regularly monitored with blood tests, urinalysis, and computed tomography. Pet owners were asked to complete monthly a quality‐of‐life questionnaire. Twenty‐four dogs were prospectively enrolled. According to Adams modified system, there were five stage 1, five stage 2, three stage 3, and 11 stage 4 tumors. Two dogs had metastases to regional lymph nodes. Median progression‐free interval and overall survival were 228 and 335 days in Group 1 (n = 12) and 234 and 244 days in Group 2 (n = 12). These differences were not statistically significant. The involvement of regional lymph nodes was significantly associated with progression‐free interval and overall survival (P = 0.004). Quality of life was significantly improved in Group 1 (P = 0.008). In particular, a significant difference was observed for activity and appetite. Although not providing a significant enhancement of progression‐free interval and overall survival, firocoxib in combination with RT is safe and improved life quality in dogs with nasal carcinomas.     

Survey of polychlorinated dibenzo-p-dioxins (PCDDs), Polychlorinated Dibenzo-p-furans (PCDFs), polychlorinated biphenyls (PCBs), and mineral components in Italian citrus cold-pressed essential oils

Investigation of PCDDs, PCDFs, PCBs, Al, As, Pb, Ba, Co, Cu, Cr, Fe, Mn, Cd, Ag, Sn, Zn, and Hg contents in 60 samples of cold-pressed essential oils produced in Calabria and Sicily in 2003-2005 was carried out. PCDDs, PCDFs, and PCBs were analyzed by HRGC-HRMS techniques using U.S. EPA 1613/94 and U.S. EPA 1668/A (1999) analytical methods. Mineral components were determined through GFAAS techniques; Hg content was determined by FI-M/H-AAS. The results of this study showed that essential oil contamination was due to a widespread pollution, typical background of rural areas, with relatively higher concentrations of PCDDs compared to PCDFs and little presence of PeCDF. Congeners OCDD, HpCDF, and OCDF were found at high concentrations. Regarding mineral components, mean values of Cr, Fe, and Ni were in agreement with data reported in the literature. Concentrations of As and Pb were below the maximum limits accepted by the current legislation. Finally, none of the samples analyzed were contaminated with Hg.     

Determination of cocoa butter equivalents in milk chocolate by triacylglycerol profiling

An analytical approach for the detection and quantification of cocoa butter equivalents (CBEs) in milk chocolate is presented. It is based on (i) a comprehensive standardized database covering the triacylglycerol composition of a wide range of authentic milk fat (n=310), cocoa butter (n=75), and CBE (n=74) samples and 947 gravimetrically prepared mixtures thereof, (ii) the availability of a certified cocoa butter reference material (IRMM-801) for calibration, (iii) an evaluation algorithm, which allows a reliable quantification of the milk fat content in chocolate fats using a simple linear regression model, (iv) a subsequent correction of triacylglycerols deriving from milk fat, (v) mathematical expressions to detect the presence of CBEs in milk chocolate, and (vi) a multivariate statistical formula to quantify the amount of CBEs in milk chocolate. The detection limit was 1% CBE in chocolate fat (0.3% CBE in milk chocolate, having a fat content of 30%). For quantification, the average error for prediction was 1.2% CBE in chocolate fat, corresponding to 0.4% in milk chocolate (fat content, 30%).     

Changes in soil carbon and nitrogen stocks after conversion of subtropical natural forest to loblolly pine plantations

Pinus plantations have increased in Brazil, and native forest areas have been converted for timber production. The clearing and the long-term loblolly pine (Pinus taeda L.) land-use effects on soil carbon and nitrogen stocks were evaluated in a natural broadleaved forest and in loblolly pine sites cultivated for 29, 35, 38 and 49 years, as well the soil contribution as ecosystem carbon pool. According to the exponential-decay model fitted to changes in carbon stock, the initial soil carbon stock of 200 Mg ha^?1 to a depth of 100 cm in the natural forest decreased by 36% over 49 years of pine cultivation (around 72.4 Mg ha^?1 of C). Around two-thirds of this decrease occurred in the top 30 cm of the soil and intensively in the first 12 years of cultivation, but slowly faded as carbon stock tended to reach a new steady state after approximately 49 years of cultivation. The soil nitrogen stock in the natural forest was 14.2 Mg ha^?1 to a depth of 100 cm and decreased by 36% over the 49 years. This decrease was linear according to the fitted model, especially in the top 30 cm where nitrogen decline was 83% and was proportionally more intense than the carbon decline. Despite the soil carbon decrease, soil remained the largest carbon reservoir in the ecosystem for the growing rotation time of loblolly pine in this region.

     

Ultrastructural Morphology and Nuclear Maturation Rates of Immature Equine Oocytes Vitrified with Different Solutions and Exposure Times

Ultrastructural morphological injuries and maturation rates were investigated in equine oocytes exposed to vitrification solutions (VS) containing synthetic ice blockers (SIBs) during different exposure times. In experiment 1, compact cumulus-oocyte complexes (COCs; n = 30) were randomly allocated to treatments: (1) fresh fixed (control); (2) VS-1 (1.4 M dimethyl sulfoxide [DMSO] + 1.8 M ethylene glycol [EG] + 1% SIB) for 3 minutes of equilibrium time and VS-2 (2.8 M DMSO + 3.6 M EG + 0.6 M sucrose + 1% SIB) for 1 minute (Eq-long); and (3) VS-1 for 1.5 minutes and VS-2 for 30 seconds (Eq-short). In experiment 2, compact (n = 248) and expanded (n = 264) COCs were evenly distributed to the following treatments: (1) immediate maturation in vitro (control); (2) vitrification using the Eq-short protocol as in experiment 1; and (3) vitrification using a stock solution containing 2.8 M formamide, 2.8 M DMSO, 2.7 M EG, 7% polyvinylpyrrolidone, and 1% SIB (Eq-short-mod). More (P < .02) oocytes with normal ultrastructural morphology were seen in fresh control and Eq-short groups than in Eq-long group. Metaphase-II (MII) rates were higher (P < .05) for oocytes with expanded cumulus than compact cumulus in the control group, and higher (P < .05) for oocytes with expanded cumulus than compact cumulus in Eq-short and Eq-short-mod groups. No difference in MII rates was detected among groups within each type of COC. In conclusion, reduction of exposure time to VS better preserved oocyte ultrastructural features, and MII rates were higher for vitrified oocytes with expanded cumulus. This study advances our knowledge on potential alternatives for vitrification of immature equine oocytes.     

Suitability of soil microbial parameters as indicators of heavy metal pollution

Several microbial parameters (microbial biomass, respiration, dehydrogenase, phosphatase, sulphatase, glucosidase, protease and urease activities) were measured in soils from five sites located in urban green areas close to roads differing in traffic density. Our aims were to evaluate the suitability of such parameters as field biomarkers of stress induced by heavy metal pollution, and to compare results obtained by single microbial parameters with results given by an index expressing the average microbial (AME) response of the microbial community. Data showed that all parameters were significantly reduced in the sites characterized by the highest load of metals in soil. Dehydrogenase, sulphatase, glucosidase activities and respiration, declined exponentially with increasing metal concentration, whereas phosphatase activity and AME decreased following a sigmoidal type relationship. In contrast, protease, urease and microbial biomass were not significantly correlated with soil metal concentration. Microbial parameters differed both in sensitivity to critical metal concentrations and in the rate of decline at increasing metal loads in soil. Due to the complex interplay of chemical, physical and biological factors which influence microbial activities and biomass, the proposed index (AME) appeared more suitable than single microbial parameters for a biomonitoring study of this type.