Aldobiouronic acid domains in Helicobacter pylori

Helicobacter pylori cell-surface glycans exert strong influences in host–microbe interplays and define the strain’s immunological signature. Envisaging the development of a carbohydrate-based vaccine against the gastroduodenal pathogen H. pylori, several clinical isolates are being screened for their cell-surface glycan profile. The present work concerns H. pylori clinical specimen PTAV79 that abundantly expressed amylose-like glycans. These polysaccharides were isolated in glycan-rich fractions resultant from phenol–water extractions and purified by Bio-Gel P2. Structural studies showed that the glycans are linked to glycerol and present aldobiouronic acid domains composed of [→3)-α-d-GlcA-(1→4)-α-d-Glc-(1→] repeating units. The amylose domains were constituted by an average of 19 Glc residues and the acidic moieties had an average number of 10 aldobiouronic acid repeating units. These polysaccharides were isolated in fractions that, although hydrophilic, were rich in stearic acid, strongly suggesting that they are present as glycerolipids anchored to cell-surface.     

Survey of Leptospira spp in pampas deer (Ozotoceros bezoarticus) in the Pantanal wetlands of the state of Mato Grosso do Sul, Brazil by serology and polymerase chain reaction

This work reports a survey of Leptospira spp in pampas deer (Ozotoceros bezoarticus) in the Pantanal wetlands of the state of Mato Grosso do Sul, Brazil by serology and polymerase chain reaction (PCR). Seventy pampas deer were captured in the dry season and surveyed using PCR, microscopic agglutination test (MAT) (n = 51) and by both techniques (n = 47). PCR detected infections in two pampas deer and MAT detected infections in three. Through sequencing and phylogenetic analyses, the PCR-amplified fragment detected in deer was identified as Leptospira interrogans. Serovars Pomona and Butembo were detected using MAT and the highest titre was 200 for serovar Pomona. Epidemiological aspects of the findings are discussed.     

Lycorine induces cell death in the amitochondriate parasite, Trichomonas vaginalis, via an alternative non-apoptotic death pathway

In this study, the mechanism of action of the pro-apoptotic alkaloid lycorine on an amitochondriate cell, the parasite Trichomonas vaginalis, was investigated. The cytotoxicity of lycorine against T. vaginalis was studied from 2.5 to 1000 μM and several important ultrastructural alterations were observed by electron microscopy. Lycorine arrested the T. vaginalis cell cycle, although no hallmarks of apoptosis, such as apoptotic bodies, were observed. Consequently, the underlying mechanism of action fails to completely fulfill the criteria for apoptosis. However, some similarities to paraptotic cell death were observed.     

Exogenous progesterone enhances ova and embryo quality following superstimulation of the first follicular wave in Nelore (Bos indicus) donors

Three experiments were conducted to evaluate the effects of exogenous progesterone on superovulatory response and ova/embryo quality in Bos indicus donors superstimulated during the first follicular wave (FFW). We hypothesized that exogenous progesterone during gonadotropin treatments would improve ova and embryo quality. In Experiment 1, 18 Nelore cows were randomly allocated to three groups: (1) FFW; (2) FFW plus a progesterone-releasing device (FFW+P4); and (3) control (E2+P4). Cows in the FFW groups were superstimulated beginning at synchronized ovulation, whereas cows in the control group were superstimulated after synchronization of follicular wave emergence with estradiol plus progesterone (E2+P4). There were no differences in mean (± SD) numbers of transferable embryos between FFW+P4 (8.0 ± 4.5) and control (6.7 ± 4.8) groups, but both were higher (P = 0.006) than the FFW group (0.2 ± 0.4). In Experiment 2, FFW and FFW+P4 were compared in 20 Nelore donors; exogenous progesterone increased the number of transferable embryos (3.9 ± 3.4 vs. 1.3 ± 4.1, P = 0.003). In Experiment 3, FFW and FFW+P4 were compared in 10 Nelore donors except that cows were slaughtered 12 h after pLH (Lutropin-V^®, Bioniche Animal Health, Belleville, ON, Canada) treatment. More mature cumulus oocyte complex (COC) (expanded cumulus cell layers) were collected in the FFW+P4 group than in the FFW group (21.8 ± 13.1 vs. 10.8±14.7; P = 0.003). In summary, superovulatory response was satisfactory when FSH (Folltropin-V^®, Bioniche Animal Health) treatment was initiated at emergence of the first follicular wave in Nelore (Bos indicus) donors, and the hypothesis that administration of exogenous progesterone during the treatment will improve oocyte and embryo quality was supported.     

Mode of action of the antiprion drugs 6AP and GA on ribosome assisted protein folding

The ribosome, the protein synthesis machinery of the cell, has also been implicated in protein folding. This activity resides within the domain V of the main RNA component of the large subunit of the ribosome. It has been shown that two antiprion drugs 6-aminophenanthridine (6AP) and Guanabenz (GA) bind to the ribosomal RNA and inhibit specifically the protein folding activity of the ribosome. Here, we have characterized with biochemical experiments, the mode of inhibition of these two drugs using ribosomes or ribosomal components active in protein folding (referred to as ’ribosomal folding modulators’ or RFMs) from both bacteria Escherichia coli and yeast Saccharomyces cerevisiae, and human carbonic anhydrase (HCA) as a sample protein. Our results indicate that 6AP and GA inhibit the protein folding activity of the ribosome by competition with the unfolded protein for binding to the ribosome. As a result, the yield of the refolded protein decreases, but the rate of its refolding remains unaffected. Further, 6AP- and GA mediated inhibition of RFM mediated refolding can be reversed by the addition of RFMs in excess. We also demonstrate with delayed addition of the ribosome and the antiprion drugs that there is a short time-span in the range of seconds within which the ribosome interacts with the unfolded protein. Thus we conclude that the protein folding activity of the ribosome is conserved from bacteria to eukaryotes and most likely the substrate for RFMs is an early refolding state of the target protein.     

Approximate likelihood calculation on a phylogeny for Bayesian estimation of divergence times

The molecular clock provides a powerful way to estimate species divergence times. If information on some species divergence times is available from the fossil or geological record, it can be used to calibrate a phylogeny and estimate divergence times for all nodes in the tree. The Bayesian method provides a natural framework to incorporate different sources of information concerning divergence times, such as information in the fossil and molecular data. Current models of sequence evolution are intractable in a Bayesian setting, and Markov chain Monte Carlo (MCMC) is used to generate the posterior distribution of divergence times and evolutionary rates. This method is computationally expensive, as it involves the repeated calculation of the likelihood function. Here, we explore the use of Taylor expansion to approximate the likelihood during MCMC iteration. The approximation is much faster than conventional likelihood calculation. However, the approximation is expected to be poor when the proposed parameters are far from the likelihood peak. We explore the use of parameter transforms (square root, logarithm, and arcsine) to improve the approximation to the likelihood curve. We found that the new methods, particularly the arcsine-based transform, provided very good approximations under relaxed clock models and also under the global clock model when the global clock is not seriously violated. The approximation is poorer for analysis under the global clock when the global clock is seriously wrong and should thus not be used. The results suggest that the approximate method may be useful for Bayesian dating analysis using large data sets.     

Aluminum exposure alters behavioral parameters and increases acetylcholinesterase activity in zebrafish <Emphasis Type="Italic">(Danio rerio)</Emphasis> brain

Aluminum is a metal that is known to impact fish species. The zebrafish has been used as an attractive model for toxicology and behavioral studies, being considered a model to study environmental exposures and human pathologies. In the present study, we have investigated the effect of aluminum exposure on brain acetylcholinesterase activity and behavioral parameters in zebrafish. In vivo exposure of zebrafish to 50 μg/L AlCl₃ for 96 h at pH 5.8 significantly increased (36%) acetylthiocholine hydrolysis in zebrafish brain. There were no changes in acetylcholinesterase (AChE) activity when fish were exposed to the same concentration of AlCl₃ at pH 6.8. In vitro concentrations of AlCl₃ varying from 50 to 250 μM increased AChE activity (28% to 33%, respectively). Moreover, we observed that animals exposed to AlCl₃ at pH 5.8 presented a significant decrease in locomotor activity, as evaluated by the number of line crossings (25%), distance traveled (14.1%), and maximum speed (24%) besides an increase in the absolute turn angle (12.7%). These results indicate that sublethal levels of aluminum might modify behavioral parameters and acetylcholinesterase activity in zebrafish brain.     

Monitoring Rhodotorula glutinis CCMI 145 physiological response and oil production growing on xylose and glucose using multi-parameter flow cytometry

Flow cytometry was used to monitor the lipid content, viability and intrinsic light scatter properties of Rhodotorula glutinis CCMI 145 cells growing on batch cultures using xylose and glucose as carbon sources. The highest lipid content was observed for cells grown on glucose, at the end of the exponential phase (17.8% w/w). The proportion of cells stained with PI attaining 77% at the end of the glucose growth. Cells growing on xylose produced a maximum lipid content of 10.6% (w/w), at the stationary phase. An increase in the proportion of cells stained with PI was observed, reaching 29% at the end of xylose growth. Changes in the side and forward light scatter detected during the yeast batch cultures supported that R. glutinis cells grown on glucose experienced harsher conditions, resulting in a high level of cytoplasmic membrane damage, which did not occur when R. glutinis cells grew on xylose.     

Sialylation regulates galectin-3/ligand interplay during mammary tumour progression--a case of targeted uncloaking

Galectin-3 is involved both in facilitating detachment of cells from primary tumour sites and favouring cancer cell adhesion and survival to anoikis in the blood stream. The mechanisms behind these apparently contradictory roles of the lectin have not yet been resolved. In order to investigate possible interplays between galectin-3 and its ligands underlying their role in the metastatic process, we examined mucin-1 (MUC1) and epidermal growth factor receptor (EGFR), well-known galectin-3 ligands, as well as galectin-3-binding site expression in a series of spontaneous canine malignant mammary tumours (CMMT) and a metastatic CMMT cell line. Despite the fact that CMMT cells expressed MUC1 and EGFR homogeneously over their plasma membrane, intravascular tumour cells, positive for galectin-3, expressed MUC1 and EGFR in a more focal membrane localization. Moreover, MUC1 overexpression in primary CMMT was present in parallel with down-regulation of galectin-3. Furthermore, in the CMT-U27 cell line, galectin-3 knock-down led to increased MUC1 expression, while MUC1 knock-down led to down-regulation of the lectin. Finally, removal of sialic acid from both CMMT and CMT-U27 xenograft samples exposed galectin-3-ligands throughout the tumour tissue, whereas these ligands were only present in galectin-3-positive invading cells in untreated samples. Interestingly indeed, we show that in vessel-invading cells, there is interaction between galectin-3 and the T antigen in vivo. We therefore hypothesized that loss of galectin-3 and sialylation-related masking of its ligands, in conjunction with their overexpression in specific tumour cell subpopulations, are crucial in regulating adhesive/de-adhesive events in the progression and invasive capacity of metastatic cells.     

An initial comparison of tubular netting versus tie–tie methods of cultivation for Kappaphycus alvarezii (Rhodophyta, Solieriaceae) on the south coast of Rio de Janeiro State, Brazil

A new cultivation technique for Kappaphycus alvarezii was used in the Brazilian southeastern coast (23°02′25″ S and 43°53′39″ W), the tubular netting on floating rafts. The tubular net technique (TN) was compared with the current method of tie–tie (TT). After 47 days, the daily growth rate (mean ± SD of TN and TT = 3.4 ± 0.7% day⁻¹), carrageenan yield (mean ± SD of TN and TT = 36 ± 1.3%), gel strength (mean ± SD of TN and TT = 730 ± 114.5 g cm2), and viscosity (mean ± SD of TN and TT = 350 ± 60.4 cP) did not differ between the two techniques (p > 0.05). The amount of time used to plant (TT = 30.2 ± 2.3 min and TN = 13.3 ± 3.4 min) and to harvest the seedlings (TT = 36.2 ± 2.7 min and TN = 17.8 ± 3.7 min) were lower in TN (p < 0.001). It is concluded that TN was more effective than TT, the cultivation management (time rates used to plant and harvest the seedlings) was 53.6% faster, no seedlings were lost, the cost was lower, and an estimated return in 1 year of nearly 20% more than that of the TT technique.