Cytotoxic Activity of Semi-Synthetic Derivatives of Elatol and Isoobtusol

In the present study, the in vitro cytotoxic effects of six semi-synthetic derivatives of elatol (1) and isoobtusol (2) were investigated. Chemical modifications were performed on the hydroxyl groups aiming to get derivatives of different polarity, namely the hemisuccinate, carbamate and sulfamate. The structural elucidation of the new derivatives was based on detailed NMR and MS spectroscopic analyses. The in vitro cytotoxicity of compounds 1 to 8 was evaluated against A459 and RD tumor cell lines with CC₅₀ values ranging from 4.93 to 41.53 µM. These results suggest that the structural modifications performed on both compounds could be considered a good strategy to obtain more active derivatives.     

Sulfated Galactan from Palisada flagellifera Inhibits Toxic Effects of Lachesis muta Snake Venom

In Brazil, snakebites are a public health problem and accidents caused by Lachesis muta have the highest mortality index. Envenomation by L. muta is characterized by systemic (hypotension, bleeding and renal failure) and local effects (necrosis, pain and edema). The treatment to reverse the evolution of all the toxic effects is performed by injection of antivenom. However, such therapy does not effectively neutralize tissue damage or any other local effect, since in most cases victims delay seeking appropriate medical care. In this way, alternative therapies are in demand, and molecules from natural sources have been exhaustively tested. In this paper, we analyzed the inhibitory effect of a sulfated galactan obtained from the red seaweed Palisada flagellifera against some toxic activities of L. muta venom. Incubation of sulfated galactan with venom resulted in inhibition of hemolysis, coagulation, proteolysis, edema and hemorrhage. Neutralization of hemorrhage was also observed when the galactan was administered after or before the venom injection; thus mimicking a real in vivo situation. Moreover, the galactan blocked the edema caused by a phospholipase A₂ isolated from the same venom. Therefore, the galactan from P. flagellifera may represent a promising tool to treat envenomation by L. muta as a coadjuvant for the conventional antivenom.     

Effects of fish oil replacement by vegetable oil blend on digestive enzymes and tissue histomorphology of European sea bass (<Emphasis Type="Italic">Dicentrarchus labrax</Emphasis>) juveniles

The impact of replacing circa 70 % fish oil (FO) by a vegetable oil (VO) blend (rapeseed, linseed, palm oils; 20:50:30) in diets for European sea bass juveniles (IBW 96 ± 0.8 g) was evaluated in terms of activities of digestive enzymes (amylase, lipase, alkaline phosphatase, trypsin and total alkaline proteases) in the anterior (AI) and posterior (PI) intestine and tissue morphology (pyloric caeca—PC, AI, PI, distal intestine—DI and liver). For that purpose, fish were fed the experimental diets for 36 days and then liver and intestine were sampled at 2, 6 and 24 h after the last meal. Alkaline protease characterization was also done in AI and PI at 6 h post-feeding. Dietary VO promoted higher alkaline phosphatase activity at 2 h post-feeding in the AI and at all sampling points in the PI. Total alkaline protease activity was higher at 6 h post-feeding in the PI of fish fed the FO diet. Identical number of bands was observed in zymograms of alkaline proteases of fish fed both diets. No alterations in the histomorphology of PC, AI, PI or DI were noticed in fish fed the VO diets, while in the liver a tendency towards increased hepatocyte vacuolization due to lipid accumulation was observed. Overall, and with the exception of a higher intestine alkaline phosphatase activity, 70 % FO replacement by a VO blend in diets for European sea bass resulted in no distinctive alterations on the postprandial pattern of digestive enzyme activities and intestine histomorphology.     

Dynamics of vanishing of maternally derived antibodies of Ungulate protoparvovirus 1 suggests an optimal age for gilts vaccination

The prevention of Ungulate protoparvovirus 1 (UPV1) infection and consequently the reproductive losses is based on vaccination of all pigs intended for breeding. As maternally derived antibodies (MDA) can interfere with the development of immunity following vaccination, it is important to know the duration of anti-UPV1 MDA to determine the optimal age for the best vaccination efficacy. To elucidate the association between dam and piglet antibody levels against UPV1 and to estimate the decrease rate of MDA, sera and colostrum of 127 gilts (before the first vaccination against UPV1; 15 days after the second vaccine dose; at farrowing; and during the second pregnancy) and sera of 276 piglets (prior to initial colostrum intake; at 7, 21, 57, 87, and 128 days-old) were tested by ELISA. Most gilts (85.8%) had anti-UPV1 antibodies before vaccination and after vaccination all were positive. At 7 days old, the majority of the piglets had anti-UPV1 antibodies, but around 57 days old, only 35.3% were positive and at 87 days old, all were negative. The estimated average half-life of MDA was 29.8 (28.8–30.9) days. A strong correlation was determined between piglet serum at 7 days old with gilt serum at farrowing time (r = 0.77, n = 248, P < 0.001) and with piglet serum at 7 days old with colostrum (r = 0.73, n = 248, P < 0.001). The MDA decreased earlier and the antibody half-life was a little longer than previously reported. Based on these findings, UPV1 vaccination can be performed earlier than usual.

     

Does orchiectomy enhance the immune-stimulatory effects of melatonin during experimental Chagas’ disease?

Melatonin has been reported to play a fundamental role in T-cell immunoregulation. Control of Trypanosoma cruzi parasitism during the acute phase of infection is considered to be critically dependent on direct macrophage activation by cytokines. The aim of this work was to evaluate the influence of exogenous melatonin treatment and the influences exerted by sexual hormones during the acute phase of the experimental Chagas' disease in rats. With melatonin treatment, orchiectomized animals (CMOR and IMOR) displayed the highest concentrations of IFN-γ and TNF-α. On the 7th day post-infection, untreated and treated orchiectomized animals (IOR and IMOR) showed an enhanced number of peritoneal macrophages. Nitric oxide levels were also increased in untreated and treated orchiectomized (IOR and IMOR) when compared to the other groups, with or without LPS. Our data suggest that melatonin therapy associated with orchiectomy induced a stimulating effect on the immune response to the parasite.     

In vitro evaluation,in vivo quantification,and microbial diversity studies of nutritional strategies for reducing enteric methane production

The main objective of the present work was to study nutritive strategies for lessening the CH₄ formation associated to ruminant tropical diets. In vitro gas production technique was used for evaluating the effect of tannin-rich plants, essential oils, and biodiesel co-products on CH₄ formation in three individual studies and a small chamber system to measure CH₄ released by sheep for in vivo studies was developed. Microbial rumen population diversity from in vitro assays was studied using qPCR. In vitro studies with tanniniferous plants, herbal plant essential oils derived from thyme, fennel, ginger, black seed, and Eucalyptus oil (EuO) added to the basal diet and cakes of oleaginous plants (cotton, palm, castor plant, turnip, and lupine), which were included in the basal diet to replace soybean meal, presented significant differences regarding fermentation gas production and CH₄ formation. In vivo assays were performed according to the results of the in vitro assays. Mimosa caesalpineaefolia, when supplemented to a basal diet (Tifton-85 hay Cynodon sp, corn grain, soybean meal, cotton seed meal, and mineral mixture) fed to adult Santa Ines sheep reduced enteric CH₄ emission but the supplementation of the basal diet with EuO did not affect (P > 0.05) methane released. Regarding the microbial studies of rumen population diversity using qPCR with DNA samples collected from the in vitro trials, the results showed shifts in microbial communities of the tannin-rich plants in relation to control plant. This research demonstrated that tannin-rich M. caesepineapholia, essential oil from eucalyptus, and biodiesel co-products either in vitro or in vivo assays showed potential to mitigate CH₄ emission in ruminants. The microbial community study suggested that the reduction in CH₄ production may be attributed to a decrease in fermentable substrate rather than to a direct effect on methanogenesis.     

Effect of Calea serrata Less. n-hexane extract on acetylcholinesterase of larvae ticks and brain Wistar rats

The stem bark of Acacia oxyphylla Graham ex Bentham is used as an anthelmintic by the natives of Mizoram (North-East India). Therefore, the aim of the study was to assess the effect of the active compound isolated from A. oxyphylla on the tegument of adult Raillietina echinobothrida and Ascaridia galli. The test parasites R. echinobothrida and A. galli were incubated in physiological buffered saline containing 0.0005, 0.001, 0.05, 0.1 and 1mg/ml of the isolated compound. The alterations in the tegument of the parasites post paralysis were examined using electron microscopes. The compound reduced the cestode's motility soon after incubation, but did not induce paralysis in the nematodes till about 11-14 h at highest concentration. The compound caused extensive digestion of cestode tegument as evident by electron microscopy. Disorganization of muscle bundles, loss of cell-cell contact, extreme vacuolization and oedema were some of the changes observed. Loss of cellular organelles combined with distortion of those present was markedly noted throughout the parasite tissue. Deformation and disorganization of epicuticle, disruption of mitochondrial and nuclear membrane were also observed in nematode exposed to the active compound of the plant. Substantial structural deformities in the treated parasites are indicative of an efficient vermicidal activity of the isolated compound against cestodes and nematodes.     

Effect of rhizobia,mycorrhizal fungi and phosphate-solubilizing microorganisms in the rhizosphere of native plants used to recover an iron ore area in Brazil

The iron-ore deposits in Minas Gerais state are concentrated in an area named Ferruginosous Quadrilateral, in which the dominant vegetation belongs to the Cerrado biome (savanna type, called campo rupestre or rupestrian field). This vegetation occurs in altitudes higher than 1000 m and is one of the most endangered biomes of the world. This study aimed to restore an area of rupestrian field used to keep iron-ore products, in which vegetation had been cut and thus resilience did not occur, worsening the low fertility of the fragile soil. Therefore, a revegetation model was proposed using two native species, Centrosema coriaceum (Leguminosae) and Tibouchina multiflora (Melastomataceae), inoculated with selected strains of arbuscular mycorrhizal fungi (AMF) and/or a rhizobium strain. After 48 months of plantation, the plant growth, survival index, physical and chemical soil properties, leaf nitrogen (N) and phosphorous (P), and P soil were evaluated. Inoculated plants benefited in all the analyzed aspects. While T. multiflora presented increased P content in leaves and soil rhizosphere only in inoculated plants, C. coriaceum showed the highest P content in both leaves and soil independently of inoculation. Although the inoculated plants presented more intense root colonization (AMF), the same mycorrhizal species were found in both inoculated and non-inoculated plants of C. coriaceum and T. multiflora. However, species of the Gigasporaceae family were favored in the C. coriaceum rhizosphere, as compared with T. multiflora. In addition, C. coriaceum was able to select in its rhizosphere a rhizobia strain efficient in P solubilization and a large and efficient population of phosphorus solubilizing microorganisms (PSM), preferentially composed of fungi. Through the similarity of their Internal Transcribed Spacer (ITS) sequences, the PSM fungi were identified as Fusarium oxysporum and Aspergillus fischeri using the BLAST database. The data presented suggest that C. coriaceum showed a strong rhizosphere effect in favor of a tripartite action of rhizobia, fungi and AMF populations as strategies to solubilize soil phosphate for survival and plant growth in the rupestrian field. Therefore, the inoculation of these microorganisms is desirable for mine recover.     

Mercury Methylation Capacity and Removal of Hg Species from Aqueous Medium by Cyanobacteria

Most technologies used for decontamination presents good results for high concentrations, but limitations for lower ones. The desirable Hg concentration in the water is extremely low because of its toxicity. The aims of this study were to evaluate inorganic mercury (Hg²⁺) and methylmercury (CH₃Hg⁺) toxicity in Nostoc paludosum, to assess the potential of this cyanobacteria strain to remove these Hg species from aqueous medium and also to investigate Hg methylation by the cyanobacteria. CH₃Hg⁺ determination was performed by gas chromatography-pyrolysis-atomic fluorescence spectrometry in cultures exposed to a concentration of 20 μg L^?1 for 30 days. Both Hg species were removed from the supernatant, ranging from 73 to 96% of Hg²⁺ and from 73 to 95% of CH₃Hg⁺. Ultrastructural Hg²⁺ effects in the cyanobacteria cells investigated by transmission electron microscopy revealed higher production of glycogen, cyanophycin, and intrathylacoidal spaces than the control group. When Hg was added to the culture in the form of CH₃Hg⁺, a decrease corresponding to approximately 60% of the initial concentration due to Hg volatilization was observed. The production of CH₃Hg⁺ by the cyanobacteria was detected in concentrations near the limit of detection (0.0025%) of the bioaccumulated THg. This is an advantage for biotechnological decontamination applications, as CH₃Hg⁺ is a very toxic specie and can be bioaccumulated and biomagnified. The results demonstrated that cyanobacteria cells are an efficient alternative to retain and/or remove Hg at low concentrations and they constitute a potential tool for a “final cleaning” of contaminated waste water.