Photo-induction of electrical current with the cyanobacteriumAgmenellum quadruplicatum PR-6

We investigated the possibility of eliciting a measurable photoinduced electrical current from the cyanobacteriumAgmenellum quadruplicatum PR-6 (Synechococcus PCC 7002). This proved virtually impossible for intact cells. However, treated PR-6 cells fixed in an alginate matrix on tin oxide as the active electrode in a three electrode electrochemical cell gave rise to a significant light response. Cell treatments involving toluene, chloroform or detergents were effective and gave current responses up to 250 nA. Drying the cyanobacterial matrix increased the current yield at least fifty-fold. These effects were observed for light wavelengths > 650 nm and were not influenced by inhibitors or enhancers of photosynthesis nor by sustained argon bubbling of the electrolyte.French pressure cell lysates facilitated distinction between two light induced current components. Lysates prepared without CaCl₂ gave current induction kinetics that were indistinguishable from those on chemically treated cells i.e. slowly rising to a stable maximum in 10–15 min. When CaCl₂ was present during lysis, a rapidly induced (<1 s) unstable component was observed. Toluenization of PR-6 either prior to or post lysing abolished the CaCl₂ related effect. CaCl₂ had no effect on current induction in strain PR-6008, which lacked the and subunits of phycocyanin and exhibited slow current induction kinetics.The observed effects are interpreted as responses of components of the photosystems of PR-6 rather than in terms of an integrated photosynthetic process.     

Use of Restored Small Wetlands by Breeding Waterfowl in Prince Edward Island, Canada

Abstract Since 1990 under the Eastern Habitat Joint Venture over 100 small wetlands have been restored in Prince Edward Island, Canada. Wetlands were restored by means of dredging accumulated sediment from erosion to emulate pre‐disturbance conditions (i.e., open water and extended hydroperiod). In 1998 and 1999 we compared waterfowl pair and brood use on 22 restored and 24 reference wetlands. More pairs and broods of Ring‐necked Ducks, Gadwall, Green‐winged Teal, and American Black Ducks used restored versus reference wetlands. In restored wetlands waterfowl pair density and species richness were positively correlated with wetland/cattail area, percent cattail cover, and close proximity to freshwater rivers. In addition, a waterfowl reproductive index was positively correlated with percent cattail cover. Green‐winged Teal pair occurrence in restored wetlands was positively correlated with greater amounts of open water and water depths. American Black Duck pairs occurred on most (86%) restored wetlands. Restored small wetlands likely served as stopover points for American Black Duck broods during overland or stream movements, whereas they likely served as a final brood‐rearing destination for Green‐winged Teal broods. We suggest that wetland restoration is a good management tool for increasing populations of Green‐winged Teal and American Black Ducks in Prince Edward Island.     

Influences of habitat features and human disturbance on use of breeding sites by a declining population of southern fur seals (Arctocephalus australis)

Southern fur seals Arctocephalus australis in Peru have declined gradually over the past decade, and declined dramatically (72%) as a result of low food availability during the severe El Niño in 1997–98. In 1999, seals abandoned some historically important breeding sites. This is particularly alarming because new sites were not colonized. Our objective was to examine how habitat features and human disturbance influenced whether sites were currently used, abandoned or apparently not used in the past by fur seals for breeding. Data were collected on 14 variables at 70 potential breeding sites at three guano reserves in Peru. Discriminant analysis revealed significant multivariate differences among sites currently used for breeding, abandoned sites and unused sites ( F =5.97, P <0.00001), and the model classified 74% of sites correctly. Currently used sites were less likely to have human disturbance and more likely to have offshore islands, stacked rocks, tide pools and abundant shade. Separate discriminant analyses for each reserve produced similar results. Habitat associated with thermoregulation (e.g. shade or pools) may be more important to fur seals in Peru, which breed at lower latitudes and are at greater risk of overheating on land than other populations. Habitat with minimized human access may be especially important to seals in small populations in which individuals may perceive themselves as more vulnerable because of decreased vigilance and dilution effects. Seals in our study selected breeding habitat with stacked rocks, which create shade and tide pools for thermoregulation and make human access difficult; but pups might suffer higher mortality in this habitat. We hypothesize that fur seals in Peru may exhibit an Allee effect, whereby suitability of habitat varies with population abundance.     

Identification of five hemoglobins in B6C3F1 mice by mass spectrometry and sequence analysis

The aim of the work is to identify and characterize the hemoglobins found in B6C3F1 mice using mass spectrometry. The primary structures are compared to those reported for BALB/c mice. Individual hemoglobin chains were isolated by reversed-phase high performance liquid chromatography (RP-HPLC). The molecular masses of the globins were determined using electrospray ionization (ESI) and matrix-assisted laser desorption ionization (MALDI). The purified globin chains were enzymatically cleaved and the resulting peptides were separated by RP-HPLC. The chains were identified by N-terminal sequencing and mass spectrometry (MALDI). Selected peptides were analysed by Edman degradation. ESI analysis indicates that B6C3F1 mice have two -globin chains (-1 and -2) and at least three β-globin chains, β-1, β-2 and β-3. This is one additional - and one additional β-globin chain than reported in the literature for BALB/c mice. Mass and sequence analysis of enzymatically generated peptides showed variations in the amino acid sequence in the -1, -2, β-2 and β-3 chains compared to the BALB/c mouse hemoglobins (, β^minor and β^major). The study showed that mass spectrometry in combination with traditional protein chemistry is able to identify and locate minor protein sequence variations.     

Isolation of carp cDNA clones,representing developmentally-regulated genes,using a subtractive-hybridization strategy

A subtractive-hybridization technique, combined with differential screenings and subsequent whole mount in situ hybridization (ISH) reactions, was used to isolate novel cDNA clones representing developmentally-regulated genes of carp. Small-scale differential screenings of an oocyte and a segmentation-stage cDNA library using oocyte-specific and segmentation stage-specific enriched probes, yielded 75 positive clones. ISH screening showed that 65% (15) of the oocyte-stage clones and 50% (26) of the segmentation-stage clones were indeed stage-specific. Partial sequence analysis suggests that approximately 65% of the 41 stage-specific clones represent novel genes. In addition, an Otxl clone was isolated. Two novel clones and the Otxl clone are of special interest for developmental studies. The clones represent genes that are locally expressed during embryonic development. The expression patterns of Otxl and one of the novel clones suggest functions in specification of the anterior-posterior axis. The three clones provide molecular markers for the study of gastrulation and the patterning of the a-p axis in teleosts.     

The Role of Reactive Oxygen Species in Inflammatory Disease: Evaluation of Methodology

The production of reactive oxidants has been implicated in the pathology of a number of inflammatory conditions, including inflamed arthritic joints. Many assays for the detection of these oxidants in diseased states have been described, but there are a number of potential pitfalls in both experimental design and the interpretation of results obtained with these techniques. Here, we describe a number of commonly used assays to detect the production of reactive oxidants and critically discuss their usefulness and limitations. We focus on the role of xanthine oxidase in reactive oxidant production in inflammatory disease.     

The NADP-linked glyceraldehyde-3-phosphate dehydrogenases of Anabaena variabilis and Synechocystis PCC 6803, which lack one of the cysteines found in the higher plant enzyme,are not reductively activated

Light activation of NADP-linked glyceraldehyde-3-P dehydrogenase involves reductive cleavage of a disulfide bond. We have proposed that the inactivating disulfide locks the two domains of the enzyme, preventing catalysis, and we have tentatively identified the two critical cysteine residues in the chloroplast enzyme (D. Li, F.J. Stevens, M. Schiffer and L.E. Anderson (1994) Biophys J. 67: 29–35). We reasoned that if activation of this enzyme involves these cysteines that enzymes lacking one or both should be active in the dark and insensitive to reductants. One of these cysteines is present in the enzymes from Anabaena variabilis and Synechocystis PCC 6803 but the other is not. Consistent with the proposed mechanism, glyceraldehyde-3-P dehydrogenase is not affected by DTT-treatment in extracts of either of these cyanobacteria. Fructosebisphosphatase is DTT-activated in extracts of both of these cyanobacteria and glucose-6-P dehydrogenase is inactivated in Synechocystis, as in higher plant chloroplasts. Apparently reductive modulation is possible in these cyanobacteria but glyceraldehyde-3-P dehydrogenase is not light activated.     

Molecular evidence for single Wolbachia infections among geographic strains of the flour beetle Tribolium confusum.

Infections with the rickettsial microorganism Wolbachia are cytoplasmically inherited and occur in a wide range of insect species and several other arthropods. Wolbachia infection often results in unidirectional cytoplasmic incompatibility (CI): crosses between infected males and uninfected females are incompatible and show a reduction of progeny or complete inviability. Unidirectional CI can also occur when males harbouring two incompatible Wolbachia strains are crossed with females infected with only one of the two strains. In the flour beetle Tribolium confusum, Wolbachia infections are of particular interest because of the severity of incompatibility. Typically, no progeny results from the incompatible cross, whereas only partial incompatibility is observed in most other hosts. Werren et al. (1995a) reported that Wolbachia infections in T. confusum consist of two bacterial strains belonging to distinct phylogenic groups, based on PCR amplification and sequence analysis of the bacterial cell division gene ftsZ. However, Fialho & Stevens (1996) showed that eight strains of T. confusum were infected with a single and common incompatibility type. Here we report analysis of the ftsZ gene by specific PCR amplification. Diagnostic restriction enzyme assays revealed no evidence of double infections in 11 geographic strains of T. confusum, including the strain examined by Werren et al. (1995a). Further, sequence analysis of the Wolbachia ftsZ gene and an internal transcribed spacer (ITS) region in two of these strains displayed no nucleotide variation or evidence of polymorphisms. Results suggest that T. confusum is infected with B-group Wolbachia only.