Choline and betaine as inducer agents of Pseudomonas aeruginosa phospholipase C activity in high phosphate medium

In Pseudomonas aeruginosa, choline or betaine employed as the sole carbon and nitrogen source in a high phosphate medium induced a phospholipase C and an acid phosphatase activity but not an alkaline phosphatase activity. The P. aeruginosa strain utilized in this work does not possess a constitutive phospholipase C, since under culture conditions identical to those utilized by other authors (J. Bacteriol. 93, 670-674 (1967) and J. Bacteriol. 150, 730-738 (1982), our phospholipase C proved to be an inorganic phosphate-repressible enzyme. These findings enable us to conclude that although the phosphate control for the synthesis of phospholipase C may exist, it is expressed only under certain favorable culture conditions.     

Identification and genetic diversity analysis of strain Pseudomonas syringae S5: A pathogen responsible for bacteriosis in Avena sativa plants

The genus Pseudomonas includes pathogenic species P. syringae, which can be found in various agricultural environments and which can affect a wide variety of plants, causing significant economic losses when the environmental conditions for its proliferation are optimal. Comprehensive characterizations of phytopathogenic bacteria belonging to the genus Pseudomonas are scarce in Argentina. In this work, the tabtoxin‐producing strain Pseudomonas S5, isolated from oat, was identi?ed as a P. syringae through biochemical tests such as the LOPAT test, and genetic tests such as the analysis of the small subunit ribosomal RNA gene (16S rRNA) sequence and repetitive elements, using BOX and ERIC primers. It was also determined that this phytopathogen is potentially capable of infecting other crops of agricultural importance for our region, such as soybean. This ability to infect different hosts gives it an adaptive advantage that allows it to endure seasonal changes in the environment where it lives. Our work contributes to the physiological classification of the phytopathogen P. syringae S5 isolated from our region, as well as to the knowledge about its range of potential hosts.     

Modeling and estimation of production rate for the production phase of non-growth-associated high cell density processes

Bioprocess and Biosystems Engineering - This paper addresses the estimation of the specific production rate of intracellular products and the modeling of the bioreactor volume dynamics in high cell...     

Comparative efficiency of algal biofilters in the removal of chromium and copper from wastewater

The performance of a macroalgae (Sargassum sp.), a laboratory-cultivated microalgae (Chlorococcum sp.) and a commercially available granulated activated carbon (GAC) for the removal of copper (Cu) and chromium (Cr) from aqueous solutions was evaluated using batch experiments. Kinetic and isotherm experiments were done at the optimal pH of 4.5 ± 0.1 for Cu (II) and 2.0 ± 0.1 for Cr (total). The equilibrium isotherms were determined and the results were analyzed using the Langmuir and Freundlich models. The best Cu removal performance was observed on Sargassum at a maximum removal of 87.3% obtained for an initial concentration of 20 mg L^?1 Cu. The maximum uptake capacities for Cu (II) were 71.4, 19.3 and 11.4 mg g^?1 of Sargassum, Chlorococcum and GAC, respectively. The biosorbents were also able to remove appreciable amounts of Cr, again with Sargassum showing maximum uptake capacity over the other materials. Kinetic studies also reveal that the removal rate is faster for both metals in Sargassum. Tests with an actual wastewater confirm the maximum uptake capacity of Cu by Sargassum. In all experiments the Sargassum biofilter outperformed GAC, which makes it a promising low-cost alternative to conventional filtration materials for wastewater treatment.     

Suppression of ovulation by a synthetic progestin in the capuchin monkey

Background  From the limited research in New World monkeys it is not clear whether they are as sensitive to the antiovulatory effects of synthetic progestins as noted in human beings. We examined whether levonorgestrel prevented ovulation in the capuchin monkey.
Methods  Cebus apella monkeys were treated orally with two doses of 2 mg of levonorgestrel, 8–9 hours apart, in four periovulatory stages assessed by laparoscopy.
Results  Levonorgestrel-induced luteinization of the follicle prevented oocyte release up to 8 hours before ovulation. Unhealthy oocytes were recovered from 46% of unruptured follicles. Luteal progesterone was reduced by 55%, 35%, and 25% according to when levonorgestrel was given −2, −1, and 0 day from estradiol peak respectively.
Conclusion  The capuchin monkey, a neotropical primate in which progesterone circulates at levels much higher than in Old World primates and human beings, is sensitive to the antiovulatory effects of synthetic progestins.     

Structure, function, and phylogeny of the mating locus in the Rhizopus oryzae complex

The Rhizopus oryzae species complex is a group of zygomycete fungi that are common, cosmopolitan saprotrophs. Some strains are used beneficially for production of Asian fermented foods but they can also act as opportunistic human pathogens. Although R. oryzae reportedly has a heterothallic (+/-) mating system, most strains have not been observed to undergo sexual reproduction and the genetic structure of its mating locus has not been characterized. Here we report on the mating behavior and genetic structure of the mating locus for 54 isolates of the R. oryzae complex. All 54 strains have a mating locus similar in overall organization to Phycomyces blakesleeanus and Mucor circinelloides (Mucoromycotina, Zygomycota). In all of these fungi, the minus (-) allele features the SexM high mobility group (HMG) gene flanked by an RNA helicase gene and a TP transporter gene (TPT). Within the R. oryzae complex, the plus (+) mating allele includes an inserted region that codes for a BTB/POZ domain gene and the SexP HMG gene. Phylogenetic analyses of multiple genes, including the mating loci (HMG, TPT, RNA helicase), ITS1-5.8S-ITS2 rDNA, RPB2, and LDH genes, identified two distinct groups of strains. These correspond to previously described sibling species R. oryzae sensu stricto and R. delemar. Within each species, discordant gene phylogenies among multiple loci suggest an outcrossing population structure. The hypothesis of random-mating is also supported by a 50:50 ratio of plus and minus mating types in both cryptic species. When crossed with tester strains of the opposite mating type, most isolates of R. delemar failed to produce zygospores, while isolates of R. oryzae produced sterile zygospores. In spite of the reluctance of most strains to mate in vitro, the conserved sex locus structure and evidence for outcrossing suggest that a normal sexual cycle occurs in both species.     

Stepwise construction of triple-helical heparin binding sites using peptide models

The specific localization of the asymmetric form of acetylcholinesterase (AChE) in neuromuscular junctions results from the interaction of its collagen-like tail with heparan sulfate proteoglycans in the synaptic basal lamina. This interaction involves two heparin binding consensus sequences of the form XBBXB, where B is a basic residue, located in the triple-helical collagen tail: GRKGR for the N-terminal site and GKRGK for the C-terminal site. To explore the basis of the higher heparin affinity seen for the C-terminal site vs. the N-terminal site, two homologous series of (Gly-Xaa-Yaa)(8) peptides were constructed to model these triple-helical binding sites. Individual tripeptide units from each heparin binding site were introduced in a stepwise fashion into a Gly-Pro-Hyp framework, until the consensus sequence and its surrounding triplets were recreated. As each additional triplet from the binding site is inserted to replace a host Gly-Pro-Hyp triplet, the triple-helix stability decreases, and the drop in thermal stability is close to that expected if each Gly-X-Y triplet contributed independently to global stability. CD spectroscopy and calorimetry show the stability of these AChE model peptides is increased by addition of heparin, confirming binding to heparin, and the lack of significant enthalpy change indicates the binding is largely electrostatic in nature. Displacement assays measure the strength of the peptide-heparin interaction, and indicate an inverse correlation between the peptide ability to bind heparin and its thermal stability. The model peptides for the C-terminal binding site show a greater heparin affinity than the peptide models for the N-terminal binding site only when residues surrounding the consensus sequence are included.     

Steroidogenic capacity and oxidative stress-related parameters in human luteal cell regression

The steroidogenic capacity and oxidative stress-related parameters of the human corpus luteum (CL) at different stages of the luteal phase were studied under basal and human chorionic gonadotropin (hCG)-stimulated conditions. Mid CL exhibited the maximal steroidogenic capacity, together with lower levels of glutathione and higher thiobarbituric acid reactants content, macrophage count, and superoxide dismutase (SOD) activity, compared to the late CL. Addition of hCG to luteal cell cultures led to a preferential increase in progesterone synthesis in the late CL compared to the mid CL, without changes in the oxidative stress-related parameters, except for the increased SOD activity found in the late CL. It is concluded that an oxidative stress condition is established in the mid CL, coinciding with the maximal steroidogenic capacity and macrophage infiltration of the organ, which be of relevance as one of the major mechanisms initiating CL involution in the human.     

The Mechanism of Cu+ Transport ATPases: INTERACTION WITH CU+ CHAPERONES AND THE ROLE OF TRANSIENT METAL-BINDING SITES*

Cu⁺-ATPases are membrane proteins that couple the hydrolysis of ATP to the efflux of cytoplasmic Cu⁺. In cells, soluble chaperone proteins bind and distribute cytoplasmic Cu⁺, delivering the ion to the transmembrane metal-binding sites in the ATPase. The structure of Legionella pneumophila Cu⁺-ATPase (Gourdon, P., Liu, X. Y., Skjørringe, T., Morth, J. P., Møller, L. B., Pedersen, B. P., and Nissen, P. (2011) Nature 475, 59–64) shows that a kinked transmembrane segment forms a “platform” exposed to the cytoplasm. In addition, neighboring invariant Met, Asp, and Glu are located at the “entrance” of the ion path. Mutations of amino acids in these regions of the Archaeoglobus fulgidus Cu⁺-ATPase CopA do not affect ATPase activity in the presence of Cu⁺ free in solution. However, Cu⁺ bound to the corresponding chaperone (CopZ) could not activate the mutated ATPases, and in parallel experiments, CopZ was unable to transfer Cu⁺ to CopA. Furthermore, mutation of a specific electronegative patch on the CopZ surface abolishes the ATPase activation and Cu⁺ transference, indicating that the region is required for the CopZ-CopA interaction. Moreover, the data suggest that the interaction is driven by the complementation of the electropositive platform in the ATPase and the electronegative Cu⁺ chaperone. This docking likely places the Cu⁺ proximal to the conserved carboxyl and thiol groups in the entrance site that induce metal release from the chaperone via ligand exchange. The initial interaction of Cu⁺ with the pump is transient because Cu⁺ is transferred from the entrance site to transmembrane metal-binding sites involved in transmembrane translocation.     

Copy-number polymorphisms: mining the tip of an iceberg

Copy-number polymorphisms (CNPs) represent a greatly underestimated aspect of human genetic variation. Recently, two landmark studies reported genome-wide analyses of CNPs in normal individuals and represent the beginning of an understanding of this type of large-scale variation. Future array-CGH-based CNP analyses should include standard criteria on a common microarray platform. It is only when parallel analyses of CNPs and SNPs are performed in an integrated format that we will obtain a global picture of our genetic diversity.