Effects of Aspergillus spp. exogenous fibrolytic enzymes on in vitro fermentation of tropical forages

BACKGROUND: Cellulose and hemicellulose are quantitatively the most important structural carbohydrates present in ruminant diets. Rumen micro-organisms produce enzymes that catalyse their hydrolysis, but the complex network formed by structural carbohydrates and lignin reduces their digestibility and restricts efficient utilisation of feeds by ruminants. This study aimed to produce two enzymatic extracts, apply them in ruminant diets to determine the best levels for ruminal digestibility and evaluate their effects on in vitro digestibility. RESULTS: In experiment 1 a two-stage in vitro technique was used to examine the effects of different enzymatic levels of Aspergillus japonicus and Aspergillus terricola on tropical forages. Enzyme addition had minor effects on corn silage at the highest enzymatic level. In experiment 2 an in vitro gas production (GP) technique was applied to determine apparent in vitro organic matter digestibility and metabolisable energy. The addition of enzymes in GP showed interesting results. Good data were obtained using sugar cane and Tifton-85 hay supplemented with extracts of A. japonicus and A. terricola respectively. CONCLUSION: Overall, the study suggests that addition of crude extracts containing exogenous fibrolytic enzymes to ruminant diets enhances the effective utilisation of ruminant feedstuffs such as forages. Copyright (c) 2012 Society of Chemical Industry

Exogenous Administration of 15d-PGJ(2)-Loaded Nanocapsules Inhibits Bone Resorption in a Mouse Periodontitis Model

The 15-deoxy-(Delta 12,14)-PG J(2) (15d-PGJ(2)) has demonstrated excellent anti-inflammatory results in different experimental models. It can be used with a polymeric nanostructure system for modified drug release, which can change the therapeutic properties of the active principle, leading to increased stability and slower/prolonged release. The aim of the current study was to test a nano-technological formulation as a carrier for 15d-PGJ(2), and to investigate the immunomodulatory effects of this formulation in a mouse periodontitis model. Poly (D, L-lactide-coglycolide) nanocapsules (NC) were used to encapsulate 15d-PGJ(2). BALB/c mice were infected on days 0, 2, and 4 with Aggregatibacter actinomycetemcomitans and divided into groups (n = 5) that were treated daily during 15 d with 1, 3, or 10 mu g/kg 15d-PGJ(2)-NC. The animals were sacrificed, the submandibular lymph nodes were removed for FACS analysis, and the jaws were analyzed for bone resorption by morphometry. Immunoinflammatory markers in the gingival tissue were analyzed by reverse transcriptase-quantitative PCR, Western blotting, or ELISA. Infected animals treated with the 15d-PGJ(2)-NC presented lower bone resorption than infected animals without treatment (p < 0.05). Furthermore, infected animals treated with 10 mu g/kg 15d-PGJ(2)-NC had a reduction of CD4(+)CD25(+)FOXP3(+) cells and CD4/CD8 ratio in the submandibular lymph node (p < 0.05). Moreover, CD55 was upregulated, whereas RANKL was downregulated in the gingival tissue of the 10 mu g/kg treated group (p < 0.05). Several proinflammatory cytokines were decreased in the group treated with 10 mu g/kg 15d-PGJ(2)-NC, and high amounts of 15d-PGJ(2) were observed in the gingiva. In conclusion, the 15d-PGJ(2)-NC formulation presented immunomodulatory effects, decreasing bone resorption and inflammatory responses in a periodontitis mouse model. The Journal of Immunology, 2012, 189: 1043-1052.

Exogenous ornithine is an effective precursor and the delta-ornithine amino transferase pathway contributes to proline accumulation under high N recycling in salt-stressed cashew leaves

The role of the delta-ornithine amino transferase (OAT) pathway in proline synthesis is still controversial and was assessed in leaves of cashew plants subjected to salinity. The activities of enzymes and the concentrations of metabolites involved in proline synthesis were examined in parallel with the capacity of exogenous ornithine and glutamate to induce proline accumulation. Proline accumulation was best correlated with OAT activity, which increased 4-fold and was paralleled by NADH oxidation coupled to the activities of OAT and Delta(1)-pyrroline-5-carboxylate reductase (P5CR), demonstrating the potential of proline synthesis via OAT/P5C. Overall, the activities of GS. GOGAT and aminating GDH remained practically unchanged under salinity. The activity of P5CR did not respond to NaCl whereas Delta(1)-pyrroline-5-carboxylate dehydrogenase was sharply repressed by salinity. We suggest that if the export of P5C from the mitochondria to the cytosol is possible, its subsequent conversion to proline by P5CR may be important. In a time-course experiment, proline accumulation was associated with disturbances in amino acid metabolism as indicated by large increases in the concentrations of ammonia, free amino acids, glutamine, arginine and ornithine. Conversely, glutamate concentrations increased moderately and only within the first 24 h. Exogenous feeding of ornithine as a precursor was very effective in inducing proline accumulation in intact plants and leaf discs, in which proline concentrations were several times higher than glutamate-fed or salt-treated plants. Our data suggest that proline accumulation might be a consequence of salt-induced increase in N recycling, resulting in increased levels of ornithine and other metabolites involved with proline synthesis and OAT activity. Under these metabolic circumstances the OAT pathway might contribute significantly to proline accumulation in salt-stressed cashew leaves. (C) 2011 Elsevier GmbH. All rights reserved.

Effects of exogenous calcium or oxalic acid on Pinus taeda treatment with the white-rot fungus Ceriporiopsis subvermispora

Pinus taeda wood chips were treated with the biopulping fungus Ceriporiopsis subvermispora in calcium-or oxalic acid-amended cultures. The secretion of hydrolytic and oxidative enzymes was inhibited only in the cultures having the highest concentration of calcium (1400 mg kg(-1) wood). Calcium decreased the availability of free oxalic acid, inhibited fungal growth, and reduced lignin mineralization and transformations. Oxalic acid amendment in the cultures was found not to affect the lignin mineralization and transformations; however, it did inhibit the depolymerization reactions detectable in the residual lignin that was retained in the biotreated wood. C. subvermispora presented catabolic activity for oxalic acid in the cultures amended with 1660 mg acid kg(-1) wood, whereas oxalic acid was synthesized when it was amended at low amounts or initially absent in the cultures. These data suggest one ideal ratio of oxalic acid in C. subvermispora cultures and indicate that its exogenous addition does not necessarily accompany the further degradation of lignin. Crown Copyright (C) 2012 Published by Elsevier Ltd. All rights reserved.

Exogenous influences on plant secondary metabolite levels

Plant secondary metabolites are a group of naturally occurring compound classes biosynthesized by differing biochemical pathways whose plant content and regulation is strongly susceptible to environmental influences and to potential herbal predators. Such abiotic and biotic factors might be specifically induced by means of various mechanisms, which create variation in the accumulation or biogenesis of secondary metabolites. Hence the dynamic aspect of bioactive compound synthesis and accumulation enables plants to communicate and react in order to overcome imminent threats. This contribution aims to review the most important mechanisms of various abiotic and biotic interactions, such as pathogenic microorganisms and herbivory, by which plants respond to exogenous influences, and will also report on time-scale variable influences on secondary metabolite profiles. Transmission of signals in plants commonly occurs by 'semiochemicals', which are comprised of terpenes, phenylpropanoids, benzenoids and other volatile compounds. Due to the important functions of volatile terpenes in communication processes of living organisms, as well as its emission susceptibility relative to exogenous influences, we also present different scenarios of concentration and emission variations. Toxic effects of plants vary depending on the level and type of secondary metabolites. In farming and cattle raising scenarios, the toxicity of plant secondary metabolites and respective concentration shifts may have severe consequences on livestock production and health, culminating in adverse effects on crop yields and/or their human consumers, or have an adverse economic impact. From a wider perspective, herbal medicines, agrochemicals or other natural products are also associated with variability in plant metabolite levels, which can impact the safety and reliable efficacy of these products. We also present typical examples of toxic plants which influence livestock production using Brazilian examples of toxicity of sapogenins and alkaloids on livestock to highlight the problem. (c) 2012 Elsevier B.V. All rights reserved.

Comparison of physical properties of two exogenous surfactants: new parameter.

The pulmonary surfactant has essential physical properties for normal lung function. The most important property is the surface tension. In this work, it was evaluated the surface tension of two commercial exogenous surfactants used in surfactant replacement therapy, poractant alfa (Curosurf, Chiesi Farmaceuticals, Italy) and beractant (Survanta, Abbott Laboratories, USA) using new parameters. A Langmuir film balance (Minitrough, KSV Instruments, Finland) was used to measure surface tension of poractant alfa and beractant samples. For both samples, we prepared a solution of 1 mg/m dissolved in chloroform (100π`), which was applied over a subphase of milli-Q water (175 ml) in the chamber of the balance. The chamber has two moving barriers that can change its surface area between a maximal value of 112.5 cm 2 , and a minimal value of 22.5 cm 2, defining a balance cycle. Each surfactant had its surface tension evaluated during 20 balance cycles for three times. Four quantities were calculated from the experiment: Minimum Surface Tension (MTS), defined as the surface tension at minimal surface area during the first cycle; Mean Work Cycle (MWC), defined as the mean hysteresis area of the measured surface tension curve of the last 16 balance cycles; Critical Active Surface Area in Compression (CASAC) or in Expansion (CASAE), defined as the maximal chamber area where the surfactant is active on the surface in compression or expansion. The t-test was applied to verify for statistical significance of the results. Comproved with the MST is the same reported in literature, the differences between MWC, CASAC, and CASAE were statistically significant (p<0.001). The MWC, CASAC and CASAE were higher for poractant alfa than for beractant. A higher MWC for poractant alfa means higher elastic recoil of the lung in comparison with beractant. Using a different methodology, our results showed that poractant alfa is probably more effective in a surfactant replacement therapy than beractant due the use of poractant alfa in relation to the use of beractant in preterm infants with Respiratory Distress Syndrome (RDS).

Mechanical Measurements Of Two Exogenous Surfactants Using A Langmuir Film Balance:A Comparative Study

Rationale: The primary function of surfactant is to reduce the surface tension at air-liquid interface. In this study, the surface tension behavior of two commercial surfactants, poractant alfa (ChiesiFarmaceuticals,ltaly) and beractant (Abbott Laboratories,USA), were evaluated,using new parameters. Methods: We used a Langmuir film balance (Minitrough,KSV lnstruments,Finland) to measure of surface tension of both poractant alfa and beractant samples. For both samples,we prepared a solution of 1 mg/mdl dissolved in chloroform. The solution (1uL) was applied over a subphase of milli-Q water (175 ml) in the chamber of the balance. The chamber has two moving barriers that can change its surface area between a maximum value of 112.5 cm2 anda minimum value of 22.5 cm2, defining a balance cycle.lhree sample's films were evaluated for each sample, during 20 balance cycles. Here quantify two new variables, which is the mean hysteresis area of the measured curve surface tension of the last 16 balance cycles,defined here as Mean Work Cycle (MWC), and the moment that the surfactant is active in the surface, this measure is defined here as Active Surface Area Critical in the compression (ASACC) and the expansion (ASACE). The test was applied to compare the statistical significance of the results.

Absence of placeholders abolishes exogenous attention effect in a peripheral cueing procedure

Most studies of exogenous visuospatial attention use placeholders indicating the regions where the stimuli appear on the screen. Preliminary results from our laboratory provided evidence that the attentional effect is more frequently observed when placeholders are used in these experimental procedures. Four experiments were carried out. Experiment 1 aimed at confirming the finding that the attentional effect of a spatially non-informative cue (S1) observed in the presence of placeholders disappears in their absence. The results confirmed this finding. Experiments 2, 3, and 4 examined several possible processes that could explain this finding. Experiment 2 investigated if the contribution of a faster disengagement of attention from the cued location or a stronger forward masking could explain the absence of attentional effect when no placeholders were used. Experiment 3 investigated if increased difficulty in discrimination of the target (S2) from S1 would favor the appearance of the attentional effect in the absence of placeholders. Experiment 4 investigated if an insufficient focusing of attention towards the cued location could explain the absence of attentional effect when no placeholders were used. The results of the three experiments indicated that placeholders act by reducing the discriminability of the S2. This would presumably lead to the adoption of an attentional set that favors the mobilization of attention by the S1