β-cyclodextrin and caffeine complexes with natural polyphenols from olive and olive oils: NMR, thermodynamic, and molecular modeling studies

Complexes of β-cyclodextrin (β-CD) and caffeine (Caf) with biophenols present in olive and olive oil (tyrosol, hydroxytyrosol, homovanillic acid, 3,4-dihydroxyphenylacetic acid, and protocatechuic acid) were investigated by NMR spectroscopy and thermodynamical-molecular dynamic studies to verify the formation of supermolecular aggregates. The obtained results indicated that the investigated biophenols form inclusion complexes with β-CD in a molar ratio of 1:1 in aqueous solution having binding constant values from 10- to 40-fold bigger than those of the corresponding complexes with Caf. Then, β-CD preferentially encloses the biophenol molecule, decreasing its bitter taste and, at the same time, preserving it against chemical and physical decomposition reactions that occur during storage.     

Copper Biosorption by Biomass of Marine Alga: Study of Equilibrium and Kinetics in Batch System and Adsorption/Desorption Cycles in Fixed Bed Column

Copper biosorption onto chemically modified biomass of marine alga Sargassum filipendula was investigated in a batch reactor and a fixed bed column. Experiments were carried out in the batch reactor to obtain kinetic and equilibrium data and to assess the copper desorption efficiency of different eluent solutions. The pseudo-first-order, pseudo-second-order, and Langmuir kinetic models were used to correlate kinetic data. The experimental data fitted well to the pseudo first order and Langmuir kinetic models. Langmuir and Freundlich models were applied to describe the equilibrium data obtained at a fixed temperature of 30°C and at pH values of 3.0, 4.0, 5.0, and 6.0. The maximum capacities of copper biosorption onto the algal biomass were 1.43, 1.59, 2.40, and 2.36 mequiv./g at pH 3.0, 4.0, 5.0, and 6.0, respectively. The efficiencies of two eluent solutions (calcium chloride and hydrochloric acid) for copper removal from the biomass were evaluated at different concentrations (0.1, 0.2, 0.5, and 1.0 mol/L). The efficiencies of the calcium chloride solutions varied from 1% to 14%, while efficiencies varying from 95% to 99% were obtained when hydrochloric acid solutions were applied. Three adsorption/desorption cycles were carried out in a fixed bed column using 0.1 mol/L hydrochloric acid as eluent solution. The results showed that an increase in the number of cycles led to a reduction in the adsorption capacity of the alga. The desorbed copper fraction presented no significant variation, remaining around 63% in the three adsorption/desorption cycles.     

Effects of remifentanil infusion regimens on cardiovascular function and responses to noxious stimulation in propofol-anesthetized cats

OBJECTIVE: To evaluate the effects of 2 remifentanil infusion regimens on cardiovascular function and responses to nociceptive stimulation in propofol-anesthetized cats. ANIMALS: 8 adult cats. PROCEDURES: On 2 occasions, cats received acepromazine followed by propofol (6 mg/kg then 0.3 mg/kg/min, i.v.) and a constant rate infusion (CRI) of remifentanil (0.2 or 0.3 microg/kg/ min, i.v.) for 90 minutes and underwent mechanical ventilation (phase I). After recording physiologic variables, an electrical stimulus (50 V; 50 Hz; 10 milliseconds) was applied to a forelimb to assess motor responses to nociceptive stimulation. After an interval (> or = 10 days), the same cats were anesthetized via administration of acepromazine and a similar infusion regimen of propofol; the remifentanil infusion rate adjustments that were required to inhibit cardiovascular responses to ovariohysterectomy were recorded (phase II). RESULTS: In phase I, heart rate and arterial pressure did not differ between remifentanil-treated groups. From 30 to 90 minutes, cats receiving 0.3 microg of remifentanil/kg/min had no response to noxious stimulation. Purposeful movement was detected more frequently in cats receiving 0.2 microg of remifentanil/kg/min. In phase II, the highest dosage (mean +/- SEM) of remifentanil that prevented cardiovascular responses was 0.23 +/- 0.01 microg/kg/min. For all experiments, mean time from infusion cessation until standing ranged from 115 to 140 minutes. CONCLUSIONS AND CLINICAL RELEVANCE: Although the lower infusion rate of remifentanil allowed ovariohysterectomy to be performed, a CRI of 0.3 microg/kg/min was necessary to prevent motor response to electrical stimulation in propofol-anesthetized cats. Recovery from anesthesia was prolonged with this technique.     

Effects of short-term light to heavy exercise on gastric ulcer development in horses and efficacy of omeprazole paste in preventing gastric ulceration

OBJECTIVE: To determine the effects of 8 days of light to heavy exercise on gastric ulcer development in horses and determine the efficacy of omeprazole paste in preventing gastric ulceration. DESIGN: Randomized, controlled, multicenter clinical trial. ANIMALS: 102 horses with normal-appearing gastric mucosa on endoscopic examination that were in light to heavy training. PROCEDURES: Horses at 4 trial locations were allocated into replicates and sham dosed orally (empty syringe) or treated with a paste formulation of omeprazole (1 mg/kg [0.45 mg/ lb], PO) once daily for 8 days. Training regimens varied among locations and included early training for western performance events; walking, trotting, and cantering in a mechanical exerciser; and race training (2 locations). Prevalences of gastric ulceration at the completion of the 8-day treatment period were compared between groups. RESULTS: At the end of the 8-day treatment period, the proportion of omeprazole-treated horses free from gastric ulceration (88%) was significantly higher than the proportion of sham-dosed horses free from gastric ulceration (27%). CONCLUSIONS AND CLINICAL RELEVANCE: Results showed that horses in light to heavy training for as short as 8 days were at risk of developing gastric ulcers and that administration of omeprazole paste decreased the incidence of gastric ulcers.     

Host/parasite relationship in the in vitro infection of rat gliocytes by Neospora caninum: evaluation of cell respiration

Neospora caninum is a protozoon that causes abortion in cattle and neuromuscular lesions in dogs, with the formation of cysts mainly in the central nervous system. Since N. caninum is an intracellular parasite with tropism for the cells of nervous system, this study evaluated the respiratory metabolism of glial cells infected by this* parasite. Glial cultures obtained from the cerebral cortex of newborn rats were kept in DMEM enriched with 10% fetal bovine serum, 1 mM pyruvic acid and 2 mM of L-glutamine. They were infected at a ratio of approximately 1:1 (cell/parasite). Oxygen consumption was evaluated by polarography in the non infected and N. caninum infected groups, 24 and 72 h following infection. Glial cell respiration after 24 and 72 h was 307.2 +/- 34.7 and 308.9 +/- 64.1 microL of oxygen per mug of total protein per minute, and 566.2 +/- 54.6 and 579 +/- 117.5 microL O2/microg of total protein/minute in the control and infected groups, respectively. These results show that N. caninum does not interfere with glial respiration in vitro.     

Leishmania-specific isotype levels and their relationship with specific cell-mediated immunity parameters in canine leishmaniasis

In the current retrospective study, Leishmania infantum-specific IgG, IgA and IgM levels were determined by ELISA in 106 untreated dogs with clinically-patent leishmaniasis (Sx) and in 171 clinically healthy dogs (Asx) from Spain to investigate the relationship between these Ig isotypes and clinical status. In addition, we studied if different Leishmania-specific humoral pattern exists between Asx dogs with and without cellular mediated immunity (CMI). Fifty-six dogs were assessed by means of lymphoproliferation assay (LPA), interferon production (IFN) and leishmanin skin test (LST), 71 dogs by means of both LPA and IFN and 44 only by LST. Both Sx and Asx dogs produced Leishmania-specific IgG, IgA and IgM antibodies, however the levels and proportion of positive dogs for each Ig isotype were significantly higher in Sx than in Asx ones (P<0.001). Analysis of immunological profiles determined for each cellular technique (positive and negative cellular response for each technique combined with positive or negative specific humoral response) showed that Asx dogs constituted a high heterogeneous group. No correlations were observed between CMI tests and specific IgG or IgM levels. However, a significant inverse correlation was demonstrated between specific IgA levels and LPA response (Spearman's r=-0.220; P=0.035). In general, the low correlation detected between CMI tests and isotype levels might indicate that the immune response is not strongly polarized in the majority of Asx dogs. Additionally, this study suggests that dogs developing T-cell response are probably able to avoid the dissemination of the parasite at least to mucosal surfaces and, as a consequence, to produce low or background specific IgA levels. Further studies are needed to investigate the relationship between specific IgA and parasite load, especially at mucosal site.     

Organic Fertilization in Traditional Mediterranean Grapevine Orchards Mediates Changes in Soil Microbial Community Structure and Enhances Soil Fertility

Soil microbial populations and their functions related to nutrient cycling contribute substantially to the regulation of soil fertility and the sustainability of agroecosystems. A field experiment was performed to assess the medium‐term effect of a mineral fertilizer and two organic fertilization systems with different nitrogen sources on the soil microbial community biomass, structure, and composition (phospholipid fatty acids, pattern, and abundance), microbial activity (basal respiration, dehydrogenase, protease, urease, β‐glucosidase, and total amount of phosphomonoesterase activities), and physical (aggregate stability) and chemical (total organic C, total N, available P and water‐soluble carbohydrates) properties in a vineyard under semiarid Mediterranean conditions after a period of 10 years. The three fertilization systems assayed were as follows: inorganic fertilization, addition of grapevine pruning with sheep manure (OPM), and addition of grapevine pruning with a legume cover crop (OPL). Both treatments, OPM and OPL, produced higher contents of total organic carbon, total N, available P, water‐soluble carbohydrates, and stable aggregates. The organic fertilization systems increased microbial biomass, shifted the structure and composition of the soil microbial community, and stimulated microbial activity, when compared with inorganic fertilization. The abundances of fungi and G+ bacteria were increased by treatments OPM and OPL, without significant differences between them. Organic and inorganic fertilization produced similar grapevine yields. The ability of the organic fertilization systems for promoting the sustainability and soil biological and chemical fertility of an agroecosystem under semiarid conditions was dependent of the organic N source. Copyright © 2016 John Wiley & Sons, Ltd.     

Does Biochar Improve Establishment of Tree Seedlings in Saline Sodic Soils?

Reforestation of saline sodic soil is increasingly undertaken as a means of reclaiming otherwise unproductive agricultural land. Currently, restoration of degraded land is limited to species with high tolerances of salinity. Biochar application has the potential to improve physical, biological and chemical properties of these soils to allow establishment of a wider range of plants. In a glasshouse trial, we applied biochar made from Acacia pycnantha (5 Mg ha⁻¹) or no biochar to either a low (EC_e 4·75 dS m⁻¹, ESP 6·9), a moderate (EC_e 27·6 dS m⁻¹, ESP 29·3) or a high (EC_e 49·4 dS m⁻¹, ESP 45·1) saline sodic soil. The regional common reforestation species Eucalyptus viminalis and Acacia mearnsii were planted as tubestock in to the soils. Early establishment indicators, including growth, plant condition and nutrition, were assessed at the end of a simulated growing season, 108 days after biochar application. Application of biochar increased height, and decreased root : shoot and the concentration of Mn, N and S in plants of E. viminalis when grown in the highly saline sodic soil. Biochar application increased the concentration of B in leaves of E. viminalis and increased the concentration of P, K and S in leaves of A. mearnsii when grown in the low saline sodic soil. The results confirm that there is potential for biochar to assist in reforestation of saline sodic soils. Copyright © 2015 John Wiley & Sons, Ltd.     

Characterization of dinucleotide and trinucleotide EST-derived microsatellites in the wheat genome

Over the past decade microsatellites or simple sequence repeats (SSRs) have attracted a considerable amount of attention from researchers. The aim of the present paper was to analyse expressed sequence tag-derived SSR (EST-SSR) marker variability in wheat and to investigate the relationships between the number and type of repeat units and the level of microsatellite polymorphism. Two hundred and forty-one new EST-SSR markers available in a public database () were characterized in eight durum wheat cultivars (Svevo, Ciccio, Primadur, Duilio, Meridiano, Claudio, Latino, Messapia), two accessions of Triticum turgidum var. dicoccoides (MG4343, MG29896), one accession of T. turgidum var. dicoccum (MG5323) and in the common wheat cv. Chinese Spring. Of these, 201 primer pairs (83.4%) amplified PCR products successfully, while the remaining 40 (16.6%) failed to amplify any product. Of the EST-SSRs analysed, 45.2% of the primer pairs amplified one or two PCR products. Multiple discrete PCR products were observed among both di- and trinucleotide EST-SSR markers (31.2 and 40.5%, respectively). Markers based on dinucleotide microsatellites were more polymorphic than those based on trinucleotide SSRs in the 12 wheat genotypes tested (68.9 and 52.7%, respectively). An average of 2.5 alleles for dinucleotide and 2.0 alleles for trinucleotide SSRs was observed. The data reported in the present work indicate the presence of a significant relationship between motif sequence types and polymorphism. The primer set based on the AG repeat motif showed the lowest percentage of polymorphism (55.0%), while the primer set based on the AC repeat motif showed t he highest percentage (85.0%). Among trinucleotide SSRs, the AGG microsatellite markers showed the highest percentage of polymorphism (70.0%), and the ACG motif the lowest value (25.0%). The characterization of these new EST-SSR markers and the results of our studyon the effect of repeat number and type of motifs could have important applications in the genetic analysis of agronomically important traits, quantitative trait locus discovery and marker-assisted selection.     

Inheritance of Aluminum Tolerance and its Effects on Grain Yield and Grain Quality in Oats (Avena Sativa L.)

Summary Aluminum toxicity due to the cation Al⁺³ is a major factor limiting yields in acid soils. Wide genetic variability to aluminum tolerance is found in oat genotypes. The objectives of this study were to determine the number of genes controlling aluminum tolerance in oats and to verify if any detrimental effects were present of the aluminum tolerance genes on grain yield and grain quality in Al⁺³free soils. Aluminum tolerance was estimated as the average regrowth of the main root after exposure to toxic levels of Al⁺³ in a hydroponic solution under controlled conditions. The number of genes controlling that trait was estimated from the distribution of the average root regrowth frequencies in a population of 333 recombinant inbred lines (RIL's) in generations F_5:6 and F_5:7. The effects on grain yield and grain quality were assessed in a subpopulation of 162 RIL's chosen based on their aluminum tolerance response. Aluminum tolerance in the evaluated population was controlled by one dominant major gene with the tolerant genotypes carying Al _a Al _a and the sensitive ones al _a al _a alleles. No detrimental effects of the Al _a allele on grain yield or grain quality were detected.Part of the Master of Science dissertation of the first author