The highly antigenic 53/25 kDa Taenia solium protein fraction with cathepsin-L like activity is present in the oncosphere/cysticercus and induces non-protective IgG antibodies in pigs

Cathepsin L-like proteases are secreted by several parasites including Taenia solium. The mechanism used by T. solium oncospheres to degrade and penetrate the intestine and infect the host is incompletely understood. It is assumed that intestinal degradation is driven by the proteolytic activity of enzymes secreted by the oncosphere. Blocking the proteolytic activity by an antibody response would prevent the oncosphere penetration and further infection. Serine and cysteine proteases including chymotrypsin, trypsin, elastase, and cathepsin L, are secreted by T. solium and Taenia saginata oncospheres when cultured in vitro, being potential vaccine candidates. However, the purification of a sufficient quantity of proteases secreted by oncospheres to conduct a vaccine trial is costly and lengthy. A 53/25 kDa cathepsin L-like fraction partially purified from T. solium cyst fluid was described previously as an important antigen for immunodiagnostics. In this study we found that this antigen is present in the T. solium oncosphere and is also secreted by the cysticercus. This protein fraction was tested for its ability to protect pigs against an oral challenge with T. solium oncospheres in a vaccine trial. IgG antibodies against the 53/25 kDa cathepsin L-like protein fraction were elicited in the vaccinated animals but did not confer protection.     

Biochar from Different Carbonaceous Waste Materials: Ecotoxicity and Effectiveness in the Sorption of Metal(loid)s

In this study, biochar produced by pyrolysis of urban pruning wood (Bpw) and sewage sludge (Bss) were characterized and investigated as adsorbents for the removal of Cu(II), Pb(II), Zn(II), and As(V) from contaminated solutions. Both types of biochars showed different physical-chemical properties and metal(loid) content. In Bss, Cu, Zn, and Pb concentrations exceeded the upper limit of the common ranges in soils. However, when they were tested for their effect on soil invertebrates, neither of the biochar was expected to exert negative effects as long as the dose applied as an amendment was ≤?4.8 t ha^?1. For an assessment of the effectiveness of biochar in the immobilization of metal(loid)s, three contaminated solutions with acidic pH and different pollutant concentrations were added to both types of biochar. Precipitation as oxy-hydroxides and the formation of complexes with active functional groups of the organic matter were the main mechanisms of metal(loid) fixation by the biochar, with increased precipitation and a rising pH. Both types of biochar were effective at immobilizing Pb and Cu, while Zn showed less effectiveness in this regard and As the least. The high P content of the biochar from sewage sludge favored Pb fixation, presumably forming complexes with phosphates, while competition between phosphate and arsenate ions decreased As adsorption by Fe compounds. The metal(loid)s immobilized by biochar from urban pruning wood were more bioavailable than those fixed by biochar from sewage sludge.     

Azospirillum brasilense Az39 and Bradyrhizobium japonicum E109, inoculated singly or in combination,promote seed germination and early seedling growth in corn (Zea mays L.) and soybean (Glycine max L.)

Inoculants are biological formulations that combine a stable microorganism population and various types of compounds produced and released during fermentation, such as phytohormones and plant growth regulators. Azospirillum brasilense strain Az39 and Brayrhizobium japonicum strain E109 were previously shown to produce indole 3-acetic acid (IAA), gibberellic acid (GA₃) and zeatin (Z). We tested the hypothesis that such compounds are responsible for early growth promotion in inoculated corn (Zea mays L.) and soybean (Glycine max L.) seedlings. Seeds were inoculated with Az39, E109, or both, and kept in a chamber at 20–30 °C under a controlled photoperiod to evaluate seed germination. To evaluate root and shoot length and dry weight, and number of nodules and percentage of nodulated seedlings, in soybean, seedlings were kept in a growth chamber for 14 days under similar photoperiod and temperature conditions. Az39 and E109, singly or in combination, showed the capacity to promote seed germination, nodule formation, and early development of corn and soybean seedlings. Both strains were able to excrete IAA, GA₃ and Z into the culture medium, at a concentration sufficient to produce morphological and physiological changes in young seed tissues.     

Role of galloylation and polymerization in cytoprotective effects of polyphenolic fractions against hydrogen peroxide insult

Byproducts and wastes generated by agricultural, food, and forestry industries contain large amounts of polyphenols, which can be potentially used as sources of natural or semisynthetic antioxidants. This study examined and compared the protection against peroxidative damage induced in erythrocytes and 3T3 cell line of polyphenolic fractions from white grape pomace, pine bark, and witch hazel bark. The work pays special attention to the different degrees of polymerization and galloylation of the extracts to contribute to the understanding of their mechanisms of action. Fractions demonstrated different protections against erythrocyte lipid peroxidation, hemolysis, and 3T3 cytotoxicity caused by H(2)O(2). Galloylation is claimed to be related to antioxidant protective capacity, and it is also responsible for the pro-oxidant effect observed at high doses. The results show that not only the percentage of galloylation but also the degree of polymerization are important modulators of their antioxidant capacity. In this sense, it is crucial that novel polyphenolic fractions were prepared attending a value of 3 for the mean degree of polymerization and did not exceed a 30% of galloylation to reach the highest antioxidant capacity with the lowest cytotoxic effects. For this reason, the grape extracts appear to be the best strategy to fight against hydrogen peroxide cell damage.     

Relationships between a proprietary index, bispectral index, and hemodynamic variables as a means for evaluating depth of anesthesia in dogs anesthetized with sevoflurane

OBJECTIVE: To evaluate relationships among various techniques for monitoring anesthetic depth in sevoflurane-anesthetized dogs undergoing orthopedic surgery. ANIMALS: 10 dogs. PROCEDURE: Dogs were medicated with acepromazine (0.05 mg/kg, IM), buprenorphine (0.01 mg/kg, IM), and atropine (0.04 mg/kg, IM). Anesthesia was induced and maintained with sevoflurane. Cardiovascular and respiratory responses were monitored. Anesthetic depth was monitored by use of the bispectral index (BIS), and a proprietary index was used to monitor activity of the autonomic nervous system. RESULTS: A significant decrease in BIS was seen after induction but concurrent changes were not observed for the other techniques. The proprietary index increased significantly after intubation, but no changes were seen for the other techniques. No significant changes were detected during incision or when higher nociceptive stimuli were applied. We did not identify a correlation between BIS and the proprietary index, the proprietary index and hemodynamic variables, or the BIS and hemodynamic variables during induction and maintenance. A significant increase in the proprietary index and BIS was detected at the time of resumption of reflexes. During anesthetic recovery, a correlation was found between the proprietary index and BIS but not between hemodynamic variables and the other techniques. CONCLUSIONS AND CLINICAL RELEVANCE: A significant increase in the proprietary index, but not the BIS or hemodynamic variables, was detected during intubation. Anesthetic induction with sevoflurane did not prevent the sympathetic stimulus attributable to tracheal intubation. Monitoring of hemodynamic variables does not provide sufficient information to allow clinicians to evaluate stress during anesthetic recovery.     

Apoptosis, inflammatory response and parasite load in skin of Leishmania (Leishmania) chagasi naturally infected dogs: A histomorphometric analysis

The skin has an important role in infection by Leishmania chagasi. Apoptosis modulates the inflammatory response acting distinctively either on the progression or regression of the lesions. The parasites interact with multiple regulatory systems inducing apoptosis in host cells, during cell invasion, stabilization and multiplication of pathogens. In this context, the aim of this study was to evaluate cell death within the inflammatory infiltrates, and to correlate these results with parasite load and clinical features of dogs naturally infected with L. chagasi. Fragments of skin pinnas (8 symptomatic+8 asymptomatic+6 negative controls) were used to characterize and measure the inflammatory response, parasite load and apoptosis. Diagnosis of canine leishmaniasis was confirmed by the detection of anti-Leishmania antibodies by IFA and ELISA in serum, direct visualization of the parasite and culture in spleen, liver, pinna, bone marrow and lymph nodes, and PCR (pinna). Histomorphometry was performed with images obtained from 20 representative histological fields in a light microscope. Ultra-thin sections were mounted over a 300 mesh grids, contrasted with 2% uranyl acetate and lead citrate and examined under a Transmission Electronic Microscopy. Amastigotes were only found in the skin of symptomatic animals (31.94±18.81). The number of foci and cellularity of the inflammatory infiltrates in symptomatic dogs were higher than in other groups and in asymptomatics were higher than in controls (p<0.05; Tukey). The average area, perimeter and extreme diameters of the inflammatory infiltrates obtained in symptomatic dogs were higher than in controls (p<0.05; Tukey). The apoptotic index was higher in symptomatic than in other groups and there was no difference between asymptomatics and controls (p<0.05; Tukey). Ultrastructurally, apoptotic cells were shrunken, with condensed nuclear chromatin and cytoplasm. Condensed nuclei were frequently fragmented. Internucleosomal DNA fragmentation occurred only in symptomatic cases. Amastigotes were observed within neutrophils and macrophages. Apoptosis is directly related to parasite load, intensity of inflammatory response and clinical manifestations in L. chagasi naturally infected dogs.     

Cloning and characterization of a plasminogen-binding enolase from the saliva of the argasid tick Ornithodoros moubata

Significant amounts of enolase have recently been found in the saliva of the argasid tick Ornithodoros moubata, raising the question as to what the function of enolase in the tick–host interface is. Enolase is a multifunctional glycolytic enzyme known to act as a plasminogen receptor on cellular surfaces, promoting fibrinolysis and extracellular matrix degradation. Fibrinolysis could be important for ticks to dissolve clots that might be formed during feeding as well as to prevent clotting of the ingested blood meal in the tick midgut. Additionally, enolase-mediated extracellular matrix degradation could contribute to the tick feeding lesion. Moreover, previous observations suggested an additional antihaemostatic role for O. moubata enolase as a P-selectin antagonist ligand. Accordingly, the aim of the present study was to investigate the potential role of the O. moubata salivary enolase as a plasminogen receptor and P-selectin ligand, and to evaluate its potential as an antigen target for anti-O. moubata vaccines. The study included the cloning, sequencing and recombinant production of the O. moubata enolase, plasminogen binding and activation assays, P-selectin binding assays, animal immunization trials, and RNAi knockdown of the enolase gene. Here we confirmed that enolase is secreted to the saliva of the tick and provide convincing evidence for a role of this salivary enolase as a plasminogen receptor, most likely stimulating host fibrinolysis and maintaining blood fluidity during tick feeding. The RNAi experiments and immunization trials indicated that enolase could be also involved in the regulation of tick reproduction, suggesting new potential control strategies. Finally, the P-selectin binding experiments demonstrated that this enolase is not a P-selectin ligand.     

Effects of stone pine (Pinus pinea L.) plantation spacing on initial growth and conelet entry into production

New Forests - Stone pine (Pinus pinea L.) is well-known for its fruits, the pine nuts, which are considered a delicacy and are the most expensive dry fruits in the world. This fruit is harvested...