ALBERT

Notes: An axenically cultured isolate of single-celled freshwater microalgae (Chlorella sp.) metabolized toxic selenate to volatile dimethylselenide at exceptionally high rates when transferred from mineral-nutrient solution to water for 24 h. The Se-volatilization rates were orders of magnitude higher than those similarly measured for wetland macroalgae and higher plants. Ninety percent of 20 µm selenate supplied to the microalgae incubated without nutrients was removed through accumulation and volatilization. Additions of 1 mm sulphate but not nitrate, inhibited Se accumulation and volatilization so that only 1.8% of the supplied selenate was removed. The microalgae cultured in nutrient solution without sulphate showed increased 35S-sulphate-transporter activity. Selenium K-edge X-ray absorption spectroscopy of selenate-treated microalgae cultured with or without mineral nutrients, showed that 87% of the selenate accumulated during 24 h was reductively metabolized to intermediate organic compounds such as selenomethionine and selenocystine. This is in complete contrast to higher plants that show very limited reduction of selenate. It appears that high rates of Se accumulation and volatilization by the sulphate-deprived microalgae resulted from reduced competition with chemically analogous sulphate ions for selenate uptake via up-regulated sulphate/selenate transporters and rapid reductive metabolism of selenate. Hyper-volatilization of selenate by microalgal cells may provide a novel detoxification response.

Notes: Two year classes of cultured male yellowtail flounder Limanda ferruginea were followed in order to examine puberty and recrudescence at the gonadal level, and to assess the impacts of culture conditions on male reproduction. Cultured males demonstrated a propensity for maturation as 1 year-old fish. Pubertal activation of testicular steroid production was associated with the appearance of primary spermatocytes, but a single case of a premeiotic endocrine activation was detected. The onset of puberty occurred during September and October, when adults were post-spawned or in early recrudescence. Once initiated, pubertal development proceeded in an uninterrupted progression to full maturity by June at 22 months of age. Spermatogenetic activity during the autumn led to a peak in the gonado-somatic index and total testicular mass by December. Milt could be first expressed from some pubertal males at this time, well in advance of the May to August spawning period in captivity. Meiotic activity could be absent from the testes by the spring, but residual areas of meiotic activity were found in pubertal males of one year class which may indicate a slower spermatogenetic rate. Androgen levels peaked during prespawning and early spawning; 11-ketotestosterone (11-KT) was the dominant androgen throughout spermatogenesis. Recrudescing 2 year-old males showed greater testicular development and androgen production. Spermatogenetic patterns were similar to those seen in pubertal males, with the distinction that recrudescing males had a more discernable mitotic phase prior to meiotic activity; mitotic and meiotic activity overlapped considerably during early puberty. Testicular development well in advance of spawning may confer an evolutionary advantage to males of this cold-water marine flatfish.

Notes: Pili of Neisseria meningitidis are a key virulence factor, being the major adhesin of this capsulate organism and contributing to specificity for the human host. Pili are post-translationally modified by addition of either an O-linked trisaccharide, Gal (β1-4) Gal (α1-3) 2,4-diacetamido-2,4,6-trideoxyhexose or an O-linked disaccharide Gal (α1,3) GlcNAc. The role of these structures in meningococcal pathogenesis has not been resolved. In previous studies we identified two separate genetic loci, pglA and pglBCD, involved in pilin glycosylation. Putative functions have been allocated to these genes; however, there are not enough genes to account for the complete biosynthesis of the described structures, suggesting additional genes remain to be identified. In addition, it is not known why some strains express the trisaccharide structure and some the disaccharide structure. In order to find additional genes involved in the biosynthesis of these structures, we used the recently published group A strain Z2491 and group B strain MC58 Neisseria meningitidis genomes and the unfinished Neisseria meningitidis group C strain FAM18 and Neisseria gonorrhoeae strain FA1090 genomes to identify novel genes involved in pilin glycosylation, based on homology to known oligosaccharide biosynthetic genes. We identified a new gene involved in pilin glycosylation designated pglE and examined four additional genes pglB/B2, pglF, pglG and pglH. A strain survey revealed that pglE and pglF were present in each strain examined. The pglG, pglH and pglB2 polymorphisms were not found in strain C311♯3 but were present in a large number of clinical isolates. Insertional mutations were constructed in pglE and pglF in N. meningitidis strain C311♯3, a strain with well-defined lipopolysaccharide (LPS) and pilin-linked glycan structures. Increased gel migration of the pilin subunit molecules of pglE and pglF mutants was observed by Western analysis, indicating truncation of the trisaccharide structure. Antisera specific for the C311♯3 trisaccharide failed to react with pilin from these pglE and pglF mutants. GC-MS analysis of the sugar composition of the pglE mutant showed a reduction in galactose compared with C311♯3 wild type. Analysis of amino acid sequence homologies has suggested specific roles for pglE and pglF in the biosynthesis of the trisaccharide structure. Further, we present evidence that pglE, which contains heptanucleotide repeats, is responsible for the phase variation between trisaccharide and disaccharide structures in strain C311♯3 and other strains. We also present evidence that pglG, pglH and pglB2 are potentially phase variable.

Notes: 1 Cardiac vagal outflow is the major factor determining the magnitude of heart rate (HR) variability analysed by traditional time and frequency domain methods. New analysis techniques, such as fractal and complexity methods, have been developed to probe non-linear features in HR behaviour that may not be detectable by traditional methods.2 We investigated the effects of vagal blockade (glycopyrrolate i.v. 5 μg kg−1 h−1 for 2 h, n = 8 vs. unmedicated control group, n = 8) and various breathing patterns (n = 12) on two non-linear measures of HR variability – detrended fluctuation analysis (DFA) and approximate entropy (ApEn) – in healthy male volunteers.3 Glycopyrrolate decreased the mean (±SD) ApEn from 1.46 ± 0.18 to 0.85 ± 0.24 (P = 0.001 in comparison with the control group), and increased the short-term (α1) and intermediate-term (α2) fractal scaling exponents of DFA, α1 from 0.96 ± 0.19 to 1.43 ± 0.29 (P = 0.003) and α2 from 1.13 ± 0.10 to 1.34 ± 0.14 (P 〈 0.001).4 Decrease in fixed respiration rate from 15 to 6 breaths min−1 increased α1 from 0.83 ± 0.25 to 1.18 ± 0.27 (P 〈 0.001), but decreased α2 from 0.88 ± 0.09 to 0.45 ± 0.17 (P 〈 0.001) and ApEn from 1.26 ± 0.12 to 1.10 ± 0.14 (P = 0.028). Rapid breathing (24 min−1) had no influence on these non-linear measures of HR variability. Hyperventilation (15 min−1, tidal volume increased voluntarily by 0.5 l) decreased α1 from 0.83 ± 0.25 to 0.66 ± 0.28 (P = 0.002) but did not affect α2 or ApEn.5 To conclude, vagal blockade alters the fractal scaling properties of R-R intervals (α1, α2) and reduces the complexity (ApEn) of HR behaviour. Both the fractal and complexity measures of HR variability can also be influenced by changes in the breathing pattern.

Notes: Weathering of soil minerals under forest seems to depend on the species present. To study the influence of tree species we placed unweathered vermiculites in the soil and assessed the impact in terms of saturation index of earth-alkaline cations and cation exchange capacity in 64 forest stands, dominated by different species and growing side by side at 20 sites on acidic soils. The vermiculites were of two types, one with a large charge and the other with a small charge. Minerals were maintained in the soil for 1 and 3 years.The minerals placed in the topsoil and in soils with low buffering capacity were more acidified and weathered than those deeper in the soil and in less acid conditions. The vermiculites were transformed into hydroxylated interlayered vermiculites, and the formation of hydroxides in the interlayer space decreased the vermiculites' cation exchange capacities. The high-charge vermiculite had a greater affinity for aluminium than the low-charge variety. The effect of tree species was significant but small compared with factors such as soil type, depth and duration of incubation. Nevertheless, we can rank the acidifying and weathering caused by the trees in the following order: Picea abies, Abies alba 〉 Pinus sylvestris, Pseudotsuga menziesii 〉 Quercus spp., Fagus sylvatica. This in situ experimental approach enabled us to study potential trends in pedogenesis in few years.

Notes: We investigated the effect of goethite and copper on almond β-glucosidase activity. The activity of β-glucosidase was found to be inhibited at dissolved copper concentrations exceeding 0.2 mm. Copper was most influential in the pH range 5–5.5, at which the enzyme activity was reduced by 50% or more at total copper concentrations of 0.2 mm compared with copper-free assays. At pH 4, the presence of 0.2 mm copper reduced the activity by 15% at most. Copper caused a shift of the pH optimum towards lower pH. Goethite did not influence β-glucosidase activity significantly, although up to 95% of the enzyme was adsorbed on its surface. The adsorption seemed to be caused principally by non-electrostatic forces which were too weak to affect the structure of the enzyme. Goethite reduced the inhibitory effect of copper because of the strong affinity of copper for goethite, as observed in batch adsorption experiments. The sorption of the enzyme on goethite was not competitive with copper at concentrations less than 0.2 mm; at larger concentrations, however, the presence of the enzyme reduced copper adsorption. The influence of copper on enzyme activity as well as the influence of copper in combination with goethite could be described with a model combining Michaelis–Menten enzyme kinetics with a simple conditionally first-order reaction law for the binding of copper by the enzyme.

Notes: Sample preparation for the study of humic acids involves a purification treatment to remove both mineral and organic components extracted along with the humic substances. The effects of HCl–HF purification on the chemical composition and structure of humic acids were studied in samples from eight different composts. Only small increases in the concentrations of C, N and H and a decrease in O were found in purified humic acids when compared with the unpurified materials, no more than 10% of average concentration. Total and phenolic acidity increased in the purified materials, whereas carboxylic and carbonyl acidity showed no statistically significant differences as a result of the purification. Gel permeation chromatography showed that only minor changes occurred in molecular size distribution of the humic acids, and the infrared spectra showed no appreciable differences between the purified humic acids and the untreated materials. We conclude that the HCl–HF treatment successfully reduced the ash contents of the humic acids without modifying their chemical composition and structure significantly.