Correlation between hydrolytic enzyme activities measured in bean seedlings afterTrichoderma koningii treatment combined with pregermination and the protective effect againstPythium splendens

An experimental protocol consisting in the colonisation of pregerminated bean seeds dressed withTrichoderma sp. was used in order to study the mechanisms correlated with the protective effect againstPythium splendens. Seed dressed with TH-11 (T. koningii) for 24 h presented a higher protective effect and a higher level of seed colonisation as compared to those dressed with TH-13 (T. longibranchiatum). The levels of seed coat colonisation by TH-11 and TH-13 was shown to be correlated with the carboxymethylcellulase activity, as measured in the seed coats retreived from germinating dressed bean seeds. The seed coat colonisation was also associated with an increased activities of endo-1,3--glucanase and endochitinase measured in seed extracts, and an inhibitory effect of seed extracts onPythium sporangia germination. Pretreatment of TH-13-dressed seeds with a commercial cellulase improved all parameters mentioned above, thus suggesting a role of cellulase activity in the colonisation process and the linked protective effect. The possible role of hydrolytic enzymes in the protective effects is discussed.     

Fate of humic acids isolated from natural humic substances

Composition of humic acids (HA) is a function of plant-derived inputs, degradation processes regulated by microorganisms, organo-mineral interactions and age. Characterization of different origin humic substances is important for evaluation of their contribution to stabile and labile carbon pool in the environment. The relative abundance of chemical components in HA isolated from soils, compost, commercial lignohumates, alginite, acadiane and lignite was studied with aim to quantify content of important biomarkers such as amino acid, lipids and polyphenols. HA were considered as a heterogeneous complex and high concentration of peptides, polyphenols and lipids was determined in acadian-HA to compare with soil-HA. Compost-HA contained much more amino acids to compare with soil-HA samples. Alginite-HA and lignite-HA were similar in biomarkers content to soil-HA. Fourier transform infrared spectroscopy confirmed that chemical composition and functional groups content differs with the origin, humification degree and the age of studied samples. Soil-HA are typically composed of a variety of ?OH, COOH?, C–O, C–H₂, (aliphatic and aromatic) groups, quinines, lignin fragments, polysaccharide, monosaccharide and proteins fragments, which are linked together by ?O?, ?NH?, ?H=, >C=O, metal ions and –S? groups. ¹³C NMR spectroscopy showed that aromatic carbon content was the highest in lignite-HA and soil-HA.     

Behavioral problems in Italian saddle horses

A survey was carried out in 25 riding centers to evaluate the prevalence of behavioral problems among saddle horses and to investigate the relationship between these and individual traits, training and management of the affected horses. Seven percent of the 650 horses examined had some kind of behavioral problems, and crib-biting was the most frequent. These behavioral problems seemed to be related to the sort of training and management. Show jumping horses tended to be affected simultaneously by more than one behavioral problem and to be more aggressive towards other horses. Although show jumpers were ridden more often, they had fewer opportunities to graze. The prevalence of behavioral problems was lower among those horses ridden Western style than among those ridden English style; the latter employs more hand to bit contact. The study also showed that horses groomed and ridden by one and the same person had more problems than horses groomed and ridden also by different people.     

Molecular Analysis of Phytophthora infestans (Mont.) de Bary from Chapingo,Mexico. Phylogeographic Referential

An Andean origin of Phytophthora infestans was originally based on written records, and recently inferred from mitochondrial and nuclear gene genealogies, which emphasize a common South American origin of the European isolates, and absence of the I-b haplotype in Mexico (recently identified in Tlaxcala, Mexico). In order to compare such results, we sequenced the nuclear genes β-tubulin, Ras, and Intron Ras, and rDNA ITS regions in 24 P. infestans genotypes isolated, identified, and selected, during three crop growing cycles in Chapingo, Mexico. The pathogen showed homology and 100 % sequence similarities to the ones reported for the Andes and Europe β-tubulin, Ras, and Intron Ras genes. Results for the ITS region were inconclusive. The molecular oomycete similarities from the three geographical regions (South America, Europe, and Mexico), and the Mexican genotypes characteristics reported in the species supports the hypothesis of the location of P. infestans diversity and a center of dispersal in the central highlands of Mexico.     

Lithothamnion muelleri Controls Inflammatory Responses,Target Organ Injury and Lethality Associated with Graft-versus-Host Disease in Mice

Lithothamnion muelleri (Hapalidiaceae) is a marine red alga, which is a member of a group of algae with anti-inflammatory, antitumor, and immunomodulatory properties. The present study evaluated the effects of treatment with Lithothamnion muelleri extract (LM) in a model of acute graft-versus-host disease (GVHD), using a model of adoptive splenocyte transfer from C57BL/6 donors into B6D2F1 recipient mice. Mice treated with LM showed reduced clinical signs of disease and mortality when compared with untreated mice. LM-treated mice had reduced tissue injury, less bacterial translocation, and decreased levels of proinflammatory cytokines and chemokines (interferon-γ (IFN-γ), tumor necrosis factor-α (TNF-α), chemokine (C-C motif) ligand 2 (CCL2), chemokine (C-C motif) ligand 3 (CCL3) and chemokine (C-C motif) ligand 5 (CCL5)). The polysaccharide-rich fraction derived from LM could inhibit leukocyte rolling and adhesion in intestinal venules, as assessed by intravital microscopy. LM treatment did not impair the beneficial effects of graft-versus-leukaemia (GVL). Altogether, our studies suggest that treatment with Lithothamnion muelleri has a potential therapeutic application in GVHD treatment.     

Anti HSV-1 Activity of Halistanol Sulfate and Halistanol Sulfate C Isolated from Brazilian Marine Sponge Petromica citrina (Demospongiae)

The n-butanol fraction (BF) obtained from the crude extract of the marine sponge Petromica citrina, the halistanol-enriched fraction (TSH fraction), and the isolated compounds halistanol sulfate (1) and halistanol sulfate C (2), were evaluated for their inhibitory effects on the replication of the Herpes Simplex Virus type 1 (HSV-1, KOS strain) by the viral plaque number reduction assay. The TSH fraction was the most effective against HSV-1 replication (SI = 15.33), whereas compounds 1 (SI = 2.46) and 2 (SI = 1.95) were less active. The most active fraction and these compounds were also assayed to determine the viral multiplication step(s) upon which they act as well as their potential synergistic effects. The anti-HSV-1 activity detected was mediated by the inhibition of virus attachment and by the penetration into Vero cells, the virucidal effect on virus particles, and by the impairment in levels of ICP27 and gD proteins of HSV-1. In summary, these results suggest that the anti-HSV-1 activity of TSH fraction detected is possibly related to the synergic effects of compounds 1 and 2.     

Relationship Between Soft Wheat Flour Physicochemical Composition and Cookie‐Making Performance

Nowadays in Argentina, cookies, crackers, and cakes are made of flour obtained from bread wheat with additives or enzymes that decrease the gluten strength but increase production costs. The present research work aims to study the relationship between flour physicochemical composition (particle size average [PSA], protein, damaged starch [DS], water soluble pentosans [WSP], total pentosans [TP], and gluten), alkaline water retention capacities behavior, solvent retention capacities profile (SRC) and cookie‐making performance in a set of 51 adapted soft wheat lines with diverse origin to identify better flour parameters for predicting cookie quality. Cookie factor (CF) values were 5.06–7.56. High and significant negative correlations between sucrose SRC (–0.68), water SRC (–0.65), carbonate SRC (–0.59), and CF were found, followed by lactic SRC that presented a low negative but significant correlation (r = –0.35). The flour components DS (r = –0.67), WSP (r = –0.49), and TP (r = –0.4) were negatively associated to CF. PSA showed a negative correlation with CF (r = –0.43). Protein and gluten were the flour components that affected cookie hardness, but no significant correlation were found with pentosan or DS content. A prediction equation for CF was developed. Sucrose SRC, PSA, and DS could be used to predict 68% of the variation in cookie diameter. The cluster analysis was conducted to assess differences in flour quality parameters among genotypes based on CF. Clusters 1 and 4 were typified by lower CF (5.70 and 5.23, respectively), higher DS, pentosan content, and SRC values. Cluster 2 with a relative good CF (6.47) and Cluster 3 with the best cookie quality, high CF (7.32) and low firmness, and the lowest DS, TP, WSP content, and sucrose SRC values.     

Effect of a size-based selection program on blood metabolites and immune response of Litopenaeus vannamei juveniles fed different dietary carbohydrate levels

Blood metabolites in wild and seventh-generation cultivated shrimp were measured to determine how size-based selection could alter the nutritional and immunological conditions of Litopenaeus vannamei. Wild L. vannamei juveniles and a sample of seventh-generation cultured shrimp were acclimated under identical conditions. During 55 days, shrimp were fed a high (HCHO: 44%) or a low (LCHO: 3%) carbohydrate diet for 55 days. Wild shrimp showed a direct relation between dietary CHO and lactate, protein and hemocyte levels indicating that dietary CHO was used for protein synthesis via transamination pathways. In seventh-generation cultured shrimp these parameters were inversely proportional to dietary CHO level, indicating the capacity to synthesize protein from dietary CHO was repressed in cultured shrimp. Farmed shrimp showed a limited capacity to respond to LCHO diets demonstrating high protein dependence in their metabolism and immune response. These results demonstrate that during size-based breeding programs other metabolic process than CHO catabolism can be selected. The incapacity of shrimp to use dietary CHO could limit protein reduction of diets and limit the efforts of the shrimp industry to be ecologically and environmentally profitable.     

Studies of enzymatic oxidation of TMP-fibers and lignin model compounds by a Laccase–Mediator-System using different 14C and 13C techniques

In this work, the results of the enzymatic oxidation of TMP-fibers (thermomechanical pulp) and a well-structured lignin model compound, the dehydropolymer (DHP), were investigated by different ¹⁴C and ¹³C methods, caused by a Laccase–Mediator-System (LMS). These methods are the nuclear magnetic resonance spectroscopy (¹³C-NMR) with DHP (unmarked) and the determination of the ¹⁴CO₂ release of ¹⁴C-marked DHP and TMP-fibers. The ¹³C-NMR measurements were chosen to analyze the structural changes of the LMS-treated DHP model compounds and TMP-fibers qualitatively and quantitatively. The data of ¹⁴CO₂ release give an explanation of the demethylation of DHP and TMP-fibers. The effect of the LMS is shown by comparing the results in respect of DHP and TMP-fibers, which were only treated with laccase and of an inactivated LMS as the control. Comparing the results of the ¹³C-NMR method, in particular the use of the Mediator during the enzymatical treatment, showed significant changes in the structure of the DHP. Also, the TMP-fibers were materially influenced by the LMS. The analysis of the ¹⁴CO₂ release data of the ¹⁴C-marked DHP and TMP-fibers revealed that the rate of ¹⁴CO₂ increases in the ¹⁴C-2 atom as well as in the O¹⁴CH₃ group within the first hour of Laccase–Mediator incubation. Therefore, the ¹⁴CO₂ release from the DHP was higher than from the TMP-fibers.     

An iridoid and a flavonoid from Sideritis lanata L

A new iridoid diglucoside, 10-O-(E)-p-coumaroylmelittoside (1) and a new flavone glucoside, O-(6'-O-Acetyl-)-beta-glucopyranosylchrysoeriol (2), together with four known flavone glycosides, were isolated from the aerial parts of Sideritis lanata. The structures of the new compounds were elucidated by spectroscopic methods, 2D NMR and MS.