Photoinhibition of photosynthesis in needles of two cypress (Cupressus sempervirens) clones

Photoinhibition of photosynthesis and photosynthetic recovery were studied in detached needles of cypress (Cupressus sempervirens L.) Clones 52 and 30 under controlled conditions of high irradiation (about 1900 micromol m(-2) s(-1) for 60 min; HL treatment), followed by 60 min in darkness. The degree of photoinhibition was determined based on the ratio of variable to maximum chlorophyll fluorescence (Fv/Fm), which is a measure of the potential efficiency of photosystem II (PSII), and on electron transport measurements. The Fv/Fm ratio declined in needles of both clones in response to the HL treatment. Minimal fluorescence (Fo) increased in HL-treated needles of both clones. The HL treatment decreased rates of whole-chain and PSII activity of isolated thylakoids more in Clone 52 than in Clone 30. In needles of both clones, PSI activity was less sensitive to photoinhibition than PSII activity. In the subsequent 60-min dark incubation, fast recovery was observed in needles of both clones, with PSII efficiencies reaching similar values to those in non-photoinhibited needles. The artificial exogenous electron donors diphenyl carbazide (DPC), hydroxylamine (NH2OH) and manganese chloride (MnCl2) failed to restore the HL-induced loss of PSII activity in needles of Clone 30, whereas DPC and NH2OH significantly restored PSII activity in photoinhibited needles of Clone 52. Quantification of the PSII reaction center protein D1 and the 33-kDa protein of the water-splitting complex following HL treatment of needles revealed pronounced differences between Clone 52 and Clone 30. The large decrease in PSII activity in HL-treated needles was caused by the marked loss of D1 protein and 33-kDa protein in Clone 30 and Clone 52, respectively.     

Challenges of thermal nociceptive threshold testing in the donkey

ObjectiveTo evaluate a thermal nociceptive threshold (TNT) testing device in the donkey, and the influence of potential confounding factors on TNTs.AnimalsTwo groups (Group 1 and Group 2) of eight castrated male donkeys aged 4–9 years, weighing 105–170 kg.MethodsTNTs were measured by heating a thermal probe on skin until an end-point behaviour (threshold temperature) or a cut-out temperature (51 °C) was reached. The withers and the dorsal aspect of the distal limb were used as sites for TNT testing. The effects on TNT of different confounding factors: the limb tested; rate of heating; and ambient temperature were evaluated. Data were analyzed using general linear models, and Mann-Whitney tests, p < 0.05 was considered significant.ResultsEnd-point behaviours (skin twitch or donkey looking at test device) when the thermal probe heated the withers were observed in approximately half of tests. TNT was (mean ± SD) 46.8 ± 2.85 °C. Subsequently the limb was evaluated as the test site in Group 1 followed by Group 2 donkeys; end-point behaviour being a foot-lift. In Group 1, 72% of tests ended in an end-point behaviour but the response rate was lower in Group 2 (20%), although TNTs were similar [(47.6 ± 3.3) and (47.3 ± 3.0) °C respectively] for responding animals. Rate of heating, ambient temperature and laterality (right or left) did not affect thresholds, but mean TNT was significantly higher in the forelimb (48.5 ± 2.8 °C) than the hind limb (47.4 ± 2.8 °C) (p = 0.012).ConclusionsWhen a thermal probe cut-out temperature of 51 °C was used in TNT testing in the donkey a high proportion of tests did not produce an identifiable end point behaviour. Higher cut-out temperatures damaged the skin. Under these conditions, thermal nociceptive threshold testing appears not be an appropriate analgesiometry technique in the donkey.Clinical relevanceTNT testing under these conditions is not suitable form of analgesiometry for donkeys.     

In vitro and in vivo stability of caffeic acid phenethyl ester, a bioactive compound of propolis

The in vitro biochemical stability of caffeic acid phenethyl ester in rat and human plasma was investigated and compared with the stability of other caffeic acid esters (chlorogenic acid and rosmarinic acid). The incubation of the compounds in rat plasma for up to 6 h showed that caffeic acid phenethyl ester, but not the other compounds, was hydrolyzed, whereas human plasma did not affect the stability of all the assayed compounds. The products in rat plasma were caffeic acid and an unknown compound, which was identified by mass spectrometry as caffeic acid ethyl ester, produced by transesterification in the presence of ethanol used as vehicle for standard compounds. Specific inhibitors of different plasma esterases allowed the identification of a carboxylesterase as the enzyme involved in the metabolism of caffeic acid phenethyl ester. The oral administration in rats of caffeic acid phenethyl ester in the presence of both ethanol and 2-(2-ethoxyethoxy)ethanol gave rise to a dramatic increase of caffeic acid, as well as low levels of caffeic acid phenethyl ester, caffeic acid ethyl ester, and caffeic acid 2-(2-ethoxyethoxy)ethyl ester, in urine collected within 24 h after treatment. These results suggest that caffeic acid phenethyl ester is hydrolyzed also in vivo to caffeic acid as the major metabolite and that its biological activities should be more properly assayed and compared with those of caffeic acid, its bioactive hydrolysis product. Moreover, alcohols should be carefully used in vivo as solvents for caffeic acid phenethyl ester, since they can give rise to new bioactive caffeic acid esters.     

The Effect of Consumption of Selenium Enriched Rye/Wheat Sourdough Bread on the Body’s Selenium Status

The potential of selenium-enriched rye/wheat sourdough bread as a route for supplementing dietary selenium intakes is reported. In addition to their normal diets, 24 female volunteers (24 to 25 years old) were fed either selenium-enriched bread or non-enriched bread each day (68.02 and 0.84 μg selenium day⁻¹ respectively) for 4 weeks. The chemical form of the selenium in the bread had been characterised using HPLC-ICP-MS, which showed that 42% of the extractable selenium was present as selenomethionine. Plasma selenium levels and plasma platelet glutathione peroxidase (GPx1) activity were measured in the volunteers’ blood over a 6-week period. A statistically significant difference (p = 0.001) was observed in the mean percentage change data, calculated from the plasma selenium level measurements for the enriched and control group, over the duration of the study. A comparable difference was not observed for the platelet GPx1 activity (p = 0.756), over the same period. Two weeks after cessation of the feeding stage, i.e., at t = 6 weeks, the mean percentage change value for the selenium plasma levels in the enriched group was still significantly elevated, suggesting that the absorbed selenium had been incorporated into the body’s selenium reserves, and was then being slowly released back into the volunteers’ blood.     

Endoscopic removal of esophageal and ruminal foreign bodies in 5 Holstein calves

Endoscopic removal of esophageal and ruminal foreign bodies was successfully performed in 5 Holstein-Friesian calves under sedation or general anesthesia by using an electrocautery snare or a wire-guided Dormi basket. This report describes the endoscopic manipulations, treatment, and outcomes of esophageal foreign body removal in these calves.     

The influence of various confounding factors on mechanical nociceptive thresholds in the donkey

ObjectiveTo evaluate a mechanical nociceptive threshold (MNT) testing device in the donkey, and to investigate the influence of potential confounders on MNTs generated.Study designProspective, randomised.AnimalsSixteen castrated male donkeys aged 4–9 years, weighing 105–170 kg.MethodsMechanical nociceptive thresholds were measured using an actuator with three pins placed on the dorsal aspect of the distal limb, connected to a force meter. The pins (surface area 15 mm²) were extruded onto the limb by pressurising an air-filled syringe, until the MNT force (when foot-lift was observed) or 25 N (cut-off force) was reached. Effect on MNT of presence of a companion donkey, the limb tested, rate of application of force, testing location, level of distraction, ambient temperature and hair cover at the test site was evaluated. Long and short-term repeatability of MNT was assessed. Data were analysed using general linear models and Mann–Whitney U tests, p < 0.05 was considered significant.ResultsIncreasing the rate of force application significantly increased the mean ± SD MNT from 9.2 ± 2.0 N when applied at 0.4 N sec^?1 to 10.6 ± 2.1 N when applied at 1.2 N sec^?1 (p = 0.001). No other factors significantly influenced MNT. Mean MNT remained stable over a 3 week period, however MNTs were significantly (p = 0.006) higher (12.8 ± 3.0 N cf 10.3 ± 1.9 N) after a 12 month interval.Conclusions and clinical relevanceWhen designing studies measuring MNT in donkeys, rate of application of force must be standardised. Donkeys’ MNTs have good short-term stability suggesting this technique is appropriate for short-term analgesiometry studies; however variability of MNTs over the long-term is greater.     

Biological and Artificial Matrixes as PAH Accumulators: An Experimental Comparative Study

In order to find out a new effective accumulator of polycyclic aromatic hydrocarbons (PAHs) useful for monitoring studies on a large scale and low costs, the accumulation capacity of both biological and artificial matrixes (mosses and polyester fibers, respectively) has been tested. For this purposes, Hypnum cupressiforme and dacron® were exposed to pollution airborne in two sites located nearby an active iron industry and in center of the town of Trieste, where high PAH pollution spots, due to vehicular traffic, are usually detected. The samplers were exposed in six sampling sessions for 21 days. The results obtained were compared with data collected by active PAH samplers, usually employed for official widespread monitoring. The level of correlation between the data sets was calculated. Furthermore, a repeatability study of data was performed. According to the results, both matrixes are good PAH accumulators, though they show different skills.     

Effects of nonesterified fatty acids and beta-hydroxybutyrate on functions of mononuclear cells obtained from ewes

OBJECTIVE: To assess the effects of nonesterified fatty acids (NEFA) and beta-hydroxybutyrate (BHBA) on functions of mononuclear cells obtained from ewes. ANIMALS: 6 Sardinian ewes. PROCEDURE: Mononuclear cells were cultured with concentrations of NEFA (0, 15.6, 31.2, 62.5, 125, 250, 500, 1,000, or 2,000 micromol/L) and BHBA (0, 0.45, 0.9, 1.8, or 3.6 mmol/L). Concentrations of NEFA and BHBA were intended to mimic those of ketotic or healthy ewes, and NEFA and BHBA were tested alone and in combination. Synthesis of DNA was stimulated by use of concanavalin A (Con A) or pokeweed-mitogen (PWM). Secretion of IgM was stimulated by use of PWM. RESULTS: Synthesis of DNA stimulated by Con A and PWM was significantly inhibited by high concentrations of NEFA (> or = 250 micromol/L) or by a combination of high concentrations of NEFA (> or = 250 micromol/L) and all concentrations of BHBA (> or = 0.45 mmol/L). In contrast, DNA synthesis was not inhibited by low concentrations of NEFA (< or = 125 micromol/L) or by a combination of low concentrations of NEFA (< or = 125 micromol/L) and the lowest concentration of BHBA (0.45 mmol/L). Secretion of IgM was significantly inhibited by all concentrations of NEFA and by all combinations of NEFA and BHBA concentrations. When used alone, none of the concentrations of BHBA inhibited DNA synthesis or IgM secretion. CONCLUSIONS AND CLINICAL RELEVANCE: Reduced immunoresponsiveness during ketosis is likely to be associated with an increase in plasma concentration of NEFA and not with an increase in plasma concentration of BH BA.