Production of phytohormones by root-associated saprophytic actinomycetes isolated from the actinorhizal plant Ochetophila trinervis

The aim of the present study was to evaluate phytohormone production by symbiotic and saprophytic actinomycetes isolated from the actinorhizal plant Ochetophila trinervis which had previously proved to stimulate nodulation by Frankia. Three saprophytic strains out of 122, isolated from the rhizosphere of this plant with multiple enzymatic activities were selected for plant growth experiments in pots: Streptomyces sp. (BCRU-MM40), Actinoplanes sp. (BCRU-ME3) and Micromonospora sp. (BCRU-MM18). For experiments, the symbiotic N₂-fixing strain Frankia (BCU110501), isolated from nodules of the same actinorhizal plant was used. Phytohormone production was evaluated in supernatant of non-inoculated and inoculated culture media in exponential growth phase. Indole 3-acetic acid (IAA) and gibberellic acid (GA₃) were analyzed by gas chromatography-mass spectrometry (GC–MS), while zeatine (Z) production was determined by gas chromatography-flame ionization detector and high performance liquid chromatography (HPLC fluorescent and UV). The levels of the three phytohormones produced by the saprophytic rhizoactinomycetes were higher than that produced by the symbiotic Frankia strain. Zeatine biosynthesis was higher (μg ml⁻¹) than IAA and GA₃ (ng ml⁻¹), and Micromonospora strain produced the highest levels of these phytohormones. Although O. trinervis has been shown to be intercellularly infected by Frankia without mediation of root hair deformation, when plants were co-inoculated with actinomycetes’ culture, some root hair deformation was observed. This is the first report on identification of IAA, GA₃ and Z in saprophytic actinomycetes and their potential role in plant–microbe interaction.     

Genetic identification of units for conservation in tomato frogs, genus Dyscophus

Dyscophus antongilii and D. guineti are two morphologically very similar microhylid frogs from Madagascar of uncertain taxonomy. D. antongilii is currently included in Appendix I of the Convention on the International Trade in Endangered Species (CITES) and its exportation is banned completely. In contrast, D. guineti does not receive any legal protection and it is regularly exported. Field data on ecology and behaviour are to a large extent lacking. Here we report on a genetic survey of D. antongilii and D. guineti using nuclear and mitochondrial DNA markers. Sequences of a fragment of 501 bp of the mitochondrial cytochrome b gene from one population of D. antongilii and two populations of D. guineti resulted in a single haplotype network, without haplotype sharing among the populations. However, haplotypes of D.␣guineti were only 1–4 mutational steps from those of D. antongilii, and did not form a clade. The analysis of eight microsatellites newly developed and standardized for D. antongilii revealed an excess of homozygotes and the absence of Hardy–Weinberg equilibrium. The microsatellite data clearly distinguished between D. antongilii and D. guineti, and fixed differences were observed at one locus. Although confirmation of the status of Dyscophus antongilii and D. guineti as separate species requires further data, our study supports the definition of these two taxa as different evolutionary significant units under the adaptive evolutionary conservation concept.     

The Immunome of Colon Cancer: Functional In Silico Analysis of Antigenic Proteins Deduced from IgG Microarray Profiling

Characterization of the colon cancer immunome and its autoantibody signature from differentially-reactive antigens (DIRAGs) could provide insights into aberrant cellular mechanisms or enriched networks associated with diseases. The purpose of this study was to characterize the antibody profile of plasma samples from 32 colorectal cancer (CRC) patients and 32 controls using proteins isolated from 15,417 human cDNA expression clones on microarrays. 671 unique DIRAGs were identified and 632 were more highly reactive in CRC samples. Bioinformatics analyses reveal that compared to control samples, the immunoproteomic IgG profiling of CRC samples is mainly associated with cell death, survival, and proliferation pathways, especially proteins involved in EIF2 and mTOR signaling. Ribosomal proteins (e.g., RPL7, RPL22, and RPL27A) and CRC-related genes such as APC, AXIN1, E2F4, MSH2, PMS2, and TP53 were highly enriched. In addition, differential pathways were observed between the CRC and control samples. Furthermore, 103 DIRAGs were reported in the SEREX antigen database, demonstrating our ability to identify known and new reactive antigens. We also found an overlap of 7 antigens with 48 “CRC genes.” These data indicate that immunomics profiling on protein microarrays is able to reveal the complexity of immune responses in cancerous diseases and faithfully reflects the underlying pathology.     

Antimicrobial and antiviral activities of an actinomycete (Streptomyces sp.) isolated from a Brazilian tropical forest soil

A promising producer of bioactive compounds isolated from a Brazilian tropical soil was tested for its range of antimicrobial activities. Strain 606, classified as Streptomyces sp., could not be identified up to species level, suggesting a possible new taxon. The supernatant and 10 extracts and fractions, obtained by extraction and chromatographic techniques, presented antimicrobial activity using antibiograms. The methanolic fraction was highly active against pathogenic bacteria, phytopathogenic fungi and the human pathogenic yeast Candida albicans. It also possessed high antiviral activity inhibiting the propagation of an acyclovir-resistant herpes simplex virus type 1 strain on HEp-2 cells at non-cytotoxic concentration. The strong cytotoxic effect suggests an antitumour action. This revised version was published online in August 2006 with corrections to the Cover Date.     

Injury of mitochondrial respiration and membrane potential during iron/ascorbate-induced peroxidation

First functional events during peroxidation in mitochondria consisted in a progressive inhibition of the phosphorylating and uncoupled respiration with succinate and glutamate/malate as substrates, whereas the resting state respiration during the same period was virtually not influenced. The membrane potential registered at a time with the respiration rates was capable of being built up for a relatively long time interval with only minor decreases, and broke down rather promptly when the active respiration was highly diminished. Inhibition of respiration proceeded mainly during the initiation phase of peroxidation. Lag phases of varied length, of malondialdehyde formation which were predominantly attributed to the iron/protein ratios correlated closely with different time intervals needed to attain maximal inhibition of respiration and decrease in glutathione. Hence, the lessening of respiration, drop of membrane potential and loss of the antioxidant, glutathione, represent early stages in the causal chain of events which precede the onset of intensive lipid peroxidation.     

The excavations of La Isabela,the first European city of the New World

In the present note the author presents history of the discovery and of the status of the excavation of the site La Isabela, the first settlement planned by Columbus in the New World during his second trip to America in the summer of 1493.     

One falls,we all fall: How boys of color develop close peer mentoring relationships

The aim of this study was to investigate the processes involved in developing close peer mentoring relationships among African American and Latino male adolescents in a school-based, group peer mentoring program. Qualitative one-on-one in-depth interviews were conducted with six school staff members, who administer the program, and 26 program participants about their experiences in the mentoring program. Data analysis revealed five program processes that led to close mentoring relationships: (a) rapport-building activities, (b) safe space, (c) mutual support, (d) group identity, and (e) trust. These processes ultimately led to bonding and friendship among program members, which further solidified the previous processes. Implications and future directions are discussed.