ALBERT

Notizen: Dihydrofolate reductase is an essential bacterial enzyme necessary for the maintenance of intracellular folate pools in a biochemically active reduced state. In this report, the Mycobacterium avium folA gene was identified by functional genetic complementation, sequenced, and expressed for the first time. It has an open reading frame of 543 bp with a G+C content of 73%. The translated polypeptide sequence shows 58% identity to the consensus sequence of the conserved regions from eight other bacterial dihydrofolate reductases. Recombinant M. avium dihydrofolate reductase was expressed actively in Escherichia coli, and SDS-PAGE analysis revealed a 20 kDa species, agreeable with that predicted from the polypeptide sequence.

Notizen: Twenty-five different Eucalyptus grandis clones were artificially wounded and inoculated with a virulent isolate of Cryphonectria cubensis. The capacity of wounds to close through callus production was correlated with the relative susceptibility of these clones to infection by C. cubensis. Clones with the greatest capacity to close wounds were those that were also most tolerant to C. cubensis infection. Those with a lower capacity to close wounds were most susceptible to Cryphonectria canker.

Notizen: O-specific polysaccharide of Proteus penneri strain 41 was studied using 1H- and 13C-NMR spectroscopy, including two-dimensional COSY, heteronuclear 13C,1H-correlation (HETCOR) and one-dimensional NOE spectroscopy, and the following structure of a non-stoichiometrically O-acetylated hexasaccharide repeating unit was established:〈displayedItem type="mathematics" xml:id="mu1" numbered="no"〉〈mediaResource alt="image" href="urn:x-wiley:09288244:FIM1:FIM_1_mu1"/〉where RGlcNAc is 2-acetamido-4-O-[(S)-1-carboxyethyl]-2-deoxyglucose.Cross-reactivity of anti-P. penneri 41 O-serum with other P. penneri strains is discussed, and a new, separate O62 serogroup is proposed which is the next Proteus O-serogroup containing P. penneri strains only.

Notizen: Abstract The Kiselyovsky subterrane is the northeastern section of the Kiselyovsko-Manominsky terrane, a distinguishable tectonic unit in the north of the Sikhote-Alin Range. The terrane has been treated as part of the accretionary wedge belonging to the Khingan-Okhotsk active continental margin, but its structure and stratigraphy have been poorly understood. This paper presents new data on the subterrane structure, lithology and radiolarian biostratigraphy. The following lithostratigraphic units are established in the terrane: a ribbon chert unit, a siliceous mudstone unit and a elastics unit. Abundant Valanginian to late Hauterivian-early Barremian radiolarian assemblages are obtained from the upper part of the chert unit in addition to the known Jurassic radiolarians. The radiolarian age of the lower part of the siliceous mudstone unit (red siliceous mudstone) is determined as early Hauterivian-early Aptian. The unit's upper part (greenish-gray siliceous mudstone and dark-gray silicified mudstone) and the clastics unit contain Albian-Cenomanian assemblages. The arrangement of the units is treated as a chert-elastics sequence, whose vertical lithologic variations indicate environmental changes from a remote ocean to a convergent margin, reflecting an oceanic plate motion towards a subduction zone. The subterrane structure is a stack of imbricated slabs composed of various lithostratigraphic units, and is complicated by folding. The structure's origin is related to subduction-accretion, which occurred in the Albian-Cenomanian. The data presented provide a unique basis for accretionary wedge terranes correlation in the circum-Japan Sea Region, and the Kiselyovsky subterrane is correlated in this study with the synchronous parts of the East Sakhalin, Hidaka and Shimanto terranes. The Albian-Cenomanian radiolarian assemblages were deposited in the Boreal realm, while Valanginian ones are Tethyan; this indicates a long oceanic plate travelling to the north. The former assemblages contain an admixture of older species, redeposited by bottom traction currents and turbidite flows in trench environments.

Notizen: To assess the usefulness of the doubled haploid (DH) method in the breeding of forage grasses, a sample of anther-derived progeny of pentaploid F1 hybrids of Festuca arundinacea × Lolium multifiorum was karyotyped using genomic in situ hybridization (GISH). The technique allowed scoring of the total number of chromosomes, the number of chromosomes contributed by each parent, and the number and positions of the Festuca-Lolium translocation breakpoints. Among 27 plants analysed, 13 belonged to three clones, effectively reducing the number of different progeny karyotyped to 17. These included 10 haploids, five doubled haploids and two plants for which the origins could not be explained. In all plants analysed, a mixture of chromosomes of both parents was present, including an average of 1.88 intergeneric translocations per plant. The translocation breakpoints were distributed along almost the entire length of the chromosome arms. Chromosome variation among androgenic progeny appeared much wider than that in the conventional backcross but low vigour and high mortality suggest that this additional variation may be difficult to exploit directly in breeding. However, a change in the pattern of recombination makes the entire genome accessible to manipulation.

Notizen: Xylem hydraulic conductivity and percentage loss of conductivity (PLC) were measured on a ring-porous (Fraxinus americana L., white ash), a diffuse porous (Acer rubrum L., red maple) and a coniferous (Picea rubens Sarg., red spruce) tree species in a temperate deciduous forest in central Massachusetts, USA. Measurements were made on current and 1-year-old branch segments in the afternoon and on the following morning. Afternoon PLC was 45 to 70% for the current year's extension growth in both white ash and red maple. Morning PLC was significantly lower (10–40%). Conductivity also varied diurnally suggesting, on average, a 50% recovery from cavitation overnight. Red spruce showed lower PLC and conductivity and a less pronounced night-time recovery. Diurnal variation in hydraulic conductivity and PLC suggests that embolism removal occurred in all three species despite the existence of tension within the xylem. Further evidence for embolism removal was observed with an in situ double-staining experiment in which dyes were fed to a transpiring branch during the late afternoon and the following morning. Examination of stem cross-sections showed that a larger number of vessels were conductive in the morning than on the preceding afternoon. Results of this study suggest that hydraulic capacity is highly dynamic and that conductivity measurements reflect a balance between two processes: cavitation and embolism removal.

Notizen: Abstract: Sediment from a microbial mat from the South-West coast of the Netherlands consumed dimethylsulfide (DMS) under oxic and anoxic conditions. From this sediment, a Gram-negative, oval DMS oxidizing bacterium, strain RB-1, was isolated. Its substrate range is typical of an obligately methylotrophic organism. Enzyme analysis revealed the presence of the ribulose monophosphate pathway for carbon assimilation, and the ability to use the linear dissimilatory pathway via formate to carbon dioxide, as well as the cyclic pathway via the ribulose monophosphate route for carbon dissimilation. 16S rRNA sequence analysis showed high similarity with species belonging to the genus Methylophaga. Because of the specific dimethylsulfide and hydrogen sulfide oxidizing capacity, the new isolate was named Methylophaga sulfidovorans.

Notizen: Abstract: A model has been developed to describe the biological conversions of five functional groups of microorganisms in a microbial mat. Microbial metabolism and transport of nutrients are the two dominant processes that were considered. The microbial activity was described with Michaelis-Menten kinetics. Transport of nutrients through the mat was described by simple transport processes. The model was validated using documented measurements, and was used to simulate the fluxes of compounds and concentrations in a microbial mat during a light period of 12 hours. Experimental data from Methylopila sulfovorans cultures (isolated from a microbial mat) were incorporated in the model in order to describe DMS oxidation under varying conditions in a mat.

Notizen: Methylophaga sulfidovorans is an obligately methylotrophic, DMS-oxidizing organism, isolated from microbial mat sediment. DMS and H2S, both present in marine microbial mats, can be used as energy sources by this organism. In batch cultures of M. sulfidovorans, sequential H2S and DMS utilization occurred. In energy-limited continuous cultures, with DMS, methanol and H2S as substrates, mixotrophic growth of M. sulfidovorans was observed, showing that at low concentrations these substrates can be used simultaneously. Oxygen and H2S uptake experiments showed that the critical concentration at which sulfide inhibition of DMS oxidation occurred was between 15 and 40 μmol l−1. Also in crude enrichments of DMS oxidizers a decrease of 50% in DMS-oxidizing capacity for about 200 μmol l−1 H2S was observed. The new physiological data obtained with the pure cultures of M. sulfidovorans were incorporated in a compartment model of a microbial mat and gave improved predictions of DMS profiles and DMS emissions from the mat, both when phototrophic activity is present (day) and when it is absent (night).

Notizen: Bacterial diversity in the water column of a freshwater lake in the Netherlands was investigated by analysis of 16S rRNA gene sequences recovered through PCR amplification from total community DNA. Among 23 unique cloned sequences, two appeared to belong to the recently described bacterial division Verrucomicrobiales. One of the two sequences was most similar to a group of environmental clones that form a distinct lineage within the division. The other sequence was divergent (less than 85% similarity) from all 16S rRNA gene sequences, both from cultivated species and from environmental clones, known in this division to date. Analysis by denaturing gradient gel electrophoresis (DGGE) and sequencing of DNA recovered through excision from the DGGE gel showed that the two sequence types were present in the lake throughout the year.

Notizen: The nucleotide sequence of the α-l-arabinofuranosidase gene arfB from Clostridium stercorarium was determined. The deduced protein has a molecular mass of 56.2 kDa with an amino terminus identical to the N-terminal sequence of the purified mature enzyme from C. stercorarium. Its sequence is homologous to arabinofuranosidases of glycosyl hydrolase family 51. Sequence alignment and cluster analysis reveal three new members of glycosyl hydrolase family 51, allowing for the definition of highly conserved regions. Two of these regions are remarkably similar to the most conserved regions within several other families of glycosyl hydrolases, which have in common a (β/α)8-barrel as the core super-secondary structure, and allow to predict the acid/base catalyst and the nucleophile of the active site.

Notizen: Abstract The mycoplasmas comprise a discrete group of microorganisms that are known to exert a range of effects upon cells derived from the immune system. Some of these interactions turn out to be immunomodulatory, such as polyclonal stimulation of T and B cells or enhancement of the cytolytic potential of macrophages, NK cells and T lymphocytes. Immunologically committed cells, when infected with mycoplasmas, can also increase the production of cytokines (IL-1, IL-2, IL-4 and IL-6), interferon (IFN)γ, tumor necrosis factor-α (TNF-α) and colony-stimulating factors (particularly GM-CSF). Moreover, mycoplasmas are potent inductors of cytokine secretion by fibroblasts in culture. Since growth factors are determinants for the activation and proliferation of immunocompetent cells in vitro, we decided to investigate if these effects are concordant with the finding of mycoplasma contamination. In order to address this question, we compared the pattern of lymphokine secretion by normal-derived human fibroblasts in culture with and without Mycoplasma spp. contamination. We found those human fibroblasts that have been contaminated with mycoplasma show production of IL-13 at the transcriptional level. This effect coincides with discrete morphological changes as compared to uncontaminated human fibroblasts. This is the first report to acknowledge that mycoplasma contamination can induce mRNA expression for IL-13 in cultured human fibroblasts.

Notizen: Research was conducted to determine the efficiency and effects of chemically treating Dogway Fork, a West Virginia stream acidified by acid precipitation. We report on the water-powered rotary drum system used to apply calcium carbonate slurry to the stream. Two companion papers cover the biological and chemical effects of this treatment. The rotary drums provided near-continuous treatment over a 4–year period. Limestone aggregate (1.3–3.8 cm) high in calcium carbonate was ground within the drums into slurry form. The relatively low cost of aggregate and its ease of storage permitted economical treatment. The system compared favorably to other types of slurry dosers. Limestone particles deposited in the stream sediment continued over time to be dissolved. They were significant in the overall dissolution efficiency of the limestone treatment. This sediment calcite also provided supplementary neutralization when high flow requirements exceeded the drum station's dosing capability.

Notizen: 1. To assess the influence of lake trophic status on the occurrence of microcystin-LR (MCLR) in the tissue of resident pulmonate snails, we sampled seven lakes of varying primary productivity in Alberta, Canada. Parameters associated with productivity were measured every 2 weeks from mid-May through mid-September 1995. Phytoplankton and resident pulmonate snails were collected and analysed for MCLR concentration via high-performance liquid chromatography.2. For all species of gastropod (i.e. Lymnaea stagnalis, Helisoma trivolvis and Physa gyrina), the concentration of MCLR in the tissue was correlated (P ≤ 0.03) with toxin in the phytoplankton, but not with extracellular aqueous microcystin (P 〉 0.28). The concentrations of toxin in the tissues of L. stagnalis and P. gyrina were also correlated with the relative abundance of Microcystis spp. (P 〈 0.01).3. Given that Microcystis spp. abundance and MCLR concentration within phytoplankton are correlated with indicators of productivity, we conclude that trophic status is important in influencing the occurrence and concentration of MCLR in pulmonate snails.

Notizen: Abstract A promoter probe shuttle vector suitable for the isolation of promoter elements from coryneform bacteria was constructed. This vector carried the neomycin phosphotransferase (NPTII) gene from transposon Tn 5 as a reporter gene, and was capable of replication in both Escherichia coli and Brevibacterium flavum. The vector was used in the construction of a B. flavum library of 899 independently isolated promoter clones. Promoters with a wide range of activities in B. flavum, including some very strong promoter elements, were isolated. Comparative analysis suggests that significant differences between B. flavum and E. coli may exist in the determinants of promoter strength.

Notizen: Abstract Amino acid consumption by Entamoeba histolytica and E. invadens has been measured in order to assess the possible roles of amino acids as energy substrates. Mixtures of amino acids enhanced the growth of the parasites in complex medium and their survival in simple medium. The consumption of several amino acids by the parasites suspended in simple media was greater when glucose was absent, suggesting that they may act as alternative energy sources. Under these conditions, asparagine was consumed extremely rapidly by E. histolytica in particular, and arginine, leucine and threonine were used greatly by both species. There was also a marked consumption of aspartate, but this occurred even when glucose was present. These five amino acids and phenylalanine were the ones consumed in greatest amounts during growth of E. histolytica in complex medium. Under the same growth conditions, E. invadens also used asparagine, arginine, leucine and threonine and in addition there was a large consumption of serine and especially glutamate. In contrast, the aspartate concentration in the complex medium increased and there was also a net increase in the concentration of some other amino acids. Alanine was produced by both species when the parasites were incubated in simple medium with glucose, and in greater amounts during growth in complex media, suggesting that it is an end product of energy metabolism. The findings provide support for the suggestion that energy generation through amino acid catabolism may be a characteristic feature of anaerobic parasitic protists.

Notizen: The degree of cholesterol oxidation in commercial sweet baked foods (biscuits and snacks) and in laboratory baked biscuits, all containing fresh or powdered eggs, was determined. 7-Ketocholesterol was used as index of cholesterol oxidation and detected by two analytical methods. The analysis of the biscuits showed higher levels of 7-ketocholesterol and a more marked oxidative instability of cholesterol when prepared with powdered eggs. The significant amounts of 7-ketocholesterol found in some samples of commercial biscuits were attributed to the use of powdered eggs. These data are of importance to industries using eggs in sweet baked products which are mainly consumed by children.

Notizen: Abstract The streptococcal transposons Tn916 and Tn925 were transferred to several strains of Leuconostoc (Ln.) oenos using the filter mating method. The insertion of both transposons into the chromosome occurred at different sites. Transconjugants of Ln. oenos carrying Tn916 could serve as donors in mating experiments with Lactococcus lactis LM2301. Further analysis of L. lactis LM2301 transconjugants showed that the insertion of the transposon Tn916 into the chromosome was site-specific. These studies establish a basis for the initiation of genetic studies in this Leuconostoc species since there are no efficient conjugal or transformation systems previously described for this microorganism.

Notizen: Newly weaned pigs were fed a basal diet containing either egg antibody against fimbriae F18 at a high or low level, control egg powder or no egg, and challenged with enterotoxigenic Escherichia coli with fimbriae F18. The challenge was repeated after termination of the antibody treatment. Antibody-containing egg powder was produced by vaccination of hens with semi-purified fimbriae of the two variants F18ab and F18ac. Pigs eating egg powder with antibody against the same fimbrial variant were fully protected, even if the vaccine for the hens was produced with a different serotype devoid of enterotoxins. The effect was dose-dependent. The high dose of antibody against the heterologous variant of fimbriae F18 reduced colonisation at a level which was not significant. Ingestion of egg antibody partially suppressed the build-up of anti-colonisation immunity. Oral application of egg antibodies offers a promising approach for the prevention of infectious diseases of the digestive tract.

Notizen: Pervasive L and L ≪ S tectonites of the Mariánské Lázně eclogites include pencil fabrics, constrictional folds and mullions, that result from the first deformation increment observed. Petrological and microstructural data suggest that these structures formed by constrictional flow at deep subduction levels after, or even during, the eclogite facies stage and before the subsequent high-pressure granulite facies overprint. Constriction at mantle depths is explained by taking into account the geometry and seismic properties of recent subduction zones. Here, constrictional strain and associated L ≫ S tectonites should be expected below the rheologycontrolled slab bend (at about 40 km depth), where dip-parallel extension and a reduced viscosity of the downgoing slab are compatible with a coaxial flow regime.

Notizen: Abstract— Multiple underloads and overloads with constant ΔK were carried out on centre-cracked tension specimens. It was found that when shear lips develop, underloads or overloads affect the crack growth rate da/dN and the subsequent retardation. The appearance of the shear lip fracture surfaces depends on the frequency. At higher frequencies a greater number of rough shear lip fracture surfaces will develop, while at lower frequencies there is a tendency towards smooth shear lips. The amount of crack closure differs in each case. It was found that the type of shear lip, rough or smooth, can be related to the effect on da/dN during and after underloads. The effect of rough and smooth shear lip growth was investigated in constant ΔK tests, performing these tests with and without crack closure.

Notizen: Exploring the possibilities for commercial aquaculture has historically concentrated on reducing production uncertainty by focusing on an animal's biology and the engineering of the production facility. However, the aquaculturist faces both production and market uncertainty, both of which may significantly affect the economic feasibility of a production facility. We developed a dynamic stochastic model to compare alternative production and marketing scenarios and used the net present value of returns to evaluate the scenarios. Based on the model and the scenarios we examined, firms producing summer flounder Paralichthys dentatus are unlikely to be profitable unless they locate near and use a source of salt-water, sell their product predominately to sushi chefs (or other premium market outlets) who desire medium-sized live fish, and engage in strategies that account for market dynamics.

Notizen: Abstract: The rates of ingestion of bacteria and of accumulation of bacterial biomass by hungry Pteridomonas danica and Paraphysomonas imperforata were measured using dual radioactive-labelled bacteria in experiments lasting 4–8 h. Pteridomonas continuously consumed 4–5 bacteria h−1 throughout experiments lasting 8 h, irrespective of bacterial concentration above a threshold of about 5 × 105 bacteria ml−1, and continued to catch bacteria even below this density. The clearance rate of about 1 nl cell−1 h−1 at higher bacterial concentrations increased three or four times as bacterial numbers fell. Paraphysomonas cells, with only half the biomass of Pteridomonas, ingested up to 10 bacteria h−1 at high bacterial concentrations, and gradually reduced the feeding rate, effectively ceasing to feed at 106 bacteria ml−1; their initial clearance rate of 1–2.5 nl cell−1 h−1 subsequently fell as low as 0.1 nl cell−1 h−1. Estimation of feeding rate by extrapolation from short-term experiments on such flagellates requires extreme caution. These flagellates, starved to levels typical of the natural environment, accumulated ingested bacterial biomass at an efficiency of between 16 and 21%, indicating that in nature they would recycle 80% or more of the nutrients contained in their food.

Notizen: Abstract: The feeding of the marine ciliate Euplotes mutabilis was studied using bacteria (Vibrio natriegens) doubly labelled with 3H-thymidine and 14C-leucine. In the presence of abundant bacteria (30 × 106 bacteria ml−1), an average Euplotes cell (initially without food vacuoles) with a protein content of 12 ng consumed 16 × 103 bacteria in the first hour and 27 × 103 bacteria over four hours, accumulating about 60% of the bacterial protein into ciliate macromolecules. Euplotes which had been starved or under-fed to reduce cell protein biomass to 7 or 9 ng consumed significantly fewer bacteria, but the gross growth efficiency for protein did not change. The rate of consumption of bacteria by large Euplotes of protein content 15 ng was initially less than that of 12 ng cells, and it decreased markedly before the end of a 4-hour experiment. Recently divided cells ingested bacteria rapidly, but showed a reduced gross growth efficiency of about 40%. At low bacterial concentrations (6 × 106 bacteria ml−1) the rates of ingestion were markedly reduced to between 〈inlineGraphic alt="inline image" href="urn:x-wiley:01686496:FEM137:FEM_137_mu1" location="equation/FEM_137_mu1.gif"/〉 and 〈inlineGraphic alt="inline image" href="urn:x-wiley:01686496:FEM137:FEM_137_mu2" location="equation/FEM_137_mu2.gif"/〉 of maximal levels; the smallest cells could not sustain feeding activity at the low prey concentration and gross growth efficiency fell from 43 to 20% during a 4-hour experiment. The strategy adopted by Euplotes in response to local fluctuations in food supply involves rapid consumption with high growth efficiency in times of plenty, but slow shrinkage without cell division to survive in times of shortage.

Notizen: Bacterivorous protozoa secrete glucosaminidase enzymes which digest peptidoglycans in bacterial cell walls. Phagotrophic protozoa store digestive enzymes in lysosomes before release into food vacuoles where digestion takes place initially at acidic pH. The quantity of glucosaminidase can be assayed by measuring the rate of formation of the fluorescent product 4-methylumbelliferone by cleavage of 4-methylumbelliferyl-n-acetyl-β-d-glucosaminide. Flagellate and ciliate cells lysed with detergent possessed glucosaminidase activity which remained steady over at least 24 h. The enzyme activity increased in direct proportion to the biomass for each species tested, although the level of activity varied in different species in a way that probably relates to the ecological strategy of the species. Heterotrophic nanoplankton sampled at eight stations in waters of widely different trophic status along an Atlantic Meridional Transect showed a very strong correlation between glucosaminidase activity and estimated biomass. We conclude that this calibrated glucosaminidase assay can provide a quick and effective method of estimating the biomass of heterotrophic nanoplankton.

Notizen: Protozoa are the main consumers of heterotrophic bacteria in aquatic habitats. The numbers of these bacteria and protozoa in oligotrophic areas of the open ocean are low, and current methods lack the sensitivity to assess rates of bacterivory in such waters. A new method is proposed for estimating bacterivory on dual radioactively labelled natural bacteria using living ambient prey bacteria and separation of predators from prey by fractionation. This approach is sufficiently sensitive to measure the consumption of less than 1% of the labelled bacteria during a 13-h incubation period. When tested on samples collected from 27 stations in mesotrophic and oligotrophic regions of the North and South Atlantic Oceans, about 17% of metabolically active bacteria were grazed per day and about 60% of consumed prey biomass labelled with 14C-leucine was retained by the predators.

Notizen: Transcription of virulence genes of Bordetella pertussis is co-ordinately regulated by the BvgA and BvgS proteins, which are members of the two-component family of bacterial signal-transduction proteins. BvgS is the transmembrane sensor and BvgA the transcriptional regulator. By gel mobility shift assays we demonstrate that phosphorylated BvgA (BvgAP) forms distinct complexes with the filamentous haemag-glutinin (PFHA) promoter DNA at different BvgAP: DNA ratios. DNase I protection analyses show that phosphorylation of BvgA not only enhances affinity of the protein for the binding sites of the PFH and bvgP1 promoters, but it extends significantly the bound region towards position -35 of these promoters. Conversely, a 10-fold higher amount of BvgAP is required for binding to a large DNA region, from -168 to -60, of the pertussis toxin (Ptox) promoter sequence. These findings suggest that the molecular interaction of BvgA P with the Ptox promoter is different from its interaction with the PFHA and bvgP1 promoters. The ß70Escherichia coli RNA polymerase (RNP) does not bind to the bug-regulated promoters. However, following the formation of a BvgAP-promoter complex, the E. coli RNP specifically recognizes and binds to the bvg-regulated promoters. Thus, BvgAP exerts its action at the level of promoter recognition by directing promoter selectivity by RNP.

Notizen: Abstract HPB12-L24 was previously described as a bifunctional histone-like and ribosomal protein in Bacillus subtilis. In order to confirm the identity of HPB12 and L24, and to study the properties of this protein, the rplX gene of B. subtilis encoding L24 has been overexpressed in Escherichia coli by an efficient protein overproduction system. A simple and rapid purification scheme using ammonium sulfate precipitation and cation-exchange chromatography is presented. 10 mg of pure L24 per g of Escherichia coli cells were obtained. The purified recombinant protein L24 is heat-stable, acid-soluble and binds preferentially supercoiled DNA like protein HPB12. These results confirm the identity of HPB12 and L24. Overexpression of rplX led to gross alterations of cell morphology and to an abnormal shape of nucleoids.

Notizen: Decreasing the lipoxygenase content of soybean (Glycine max (L.) Merr.) seed is a prerequisite for its increased use in human foods. These investigations aim to analyse the variation of lipoxygenase contents in relation to the genotype or the nitrogen nutrition of the plant. Four genotypes from a divergent selection for N2 fixation and the recurrent parent, Weber, were grown in pots, and two Bradyrhizobium japonicum strains (G.49 and SMGS1) were used. Dinitrogen fixation was estimated in situ using the acetylene–reduction assay. The lipoxygenase activity was determined polarographically on seeds taken from plants grown under controlled conditions in the presence or absence of symbiotic N2 fixation. The large genotypic variation observed for the lipoxygenase contents was enhanced by high N2 fixation. The variation range of lipoxygenase contents, which was obtained through the combination of the genotypic variation and the N2 fixation activity, was relatively large with a ratio of 2 on a seed dry–weight basis and a ratio of 2.6 when the total proteins were considered. Decreased lipoxygenase content and increased total protein content can thus be considered in parallel in a genetic context, together with more efficient N2 fixation.

Notizen: The state-space estimation technique presented herein provides a method for obtaining optimal estimates of concentrations for two-dimensional plumes in ground water. The concentration of a plume was defined as the state variable. The technique uses the Kalman filter and involves combining two independent estimates of plume concentrations. One estimate is called the process modeling data and the other is called the “measurement” data. The process modeling data is obtained from a numerical model. The “measurement” data is obtained from field measurements; however, for illustration in this paper it was generated by a different numerical model than the one used to obtain the process modeling data. The state-space technique produces a distribution of contaminant concentrations that is more accurate than either of the distributions generated by the process modeling or the “measurement” data. An example is presented to show that the technique produces significant improvements in the prediction of plume concentration distributions.

Notizen: First-order kinetics with respect to the αs1-casein concentration was used to study casein degradation during low-fat Fynbo cheese ripening. Effects of partial NaCl replacement by KCI during cheese salting were studied by statistical treatment of the casein degradation results. Four zones from cheeses at 1, 5, 10, 20, and 30 ripening days were analyzed by a polyacrylamide gel electrophoresis method. Similar kinetic parameters were obtained for a cheese salted with a NaCl/KCl brine and for a control cheese during ripening. Results were more affected by salt concentration than by salt substitution. KCl did not strongly influence kinetics of Fynbo cheese proteolysis.

Notizen: In vitro culturing of plant cells can cause changes in karyotype. Chromosome variations following long-term propagation in suspension culture of Triticum timopheevii (Zhuk.) Zhuk. were studied by routine staining and C-banding. The culture was highly heterogeneous with respect to the number and structure of chromosomes. The modal class cells had a lower chromosome number than T. timopheevii (2n= 28). This data was confirmed by cytophotometric analysis of nuclear-DNA content. Frequencies of chromosome loss varied for different homoeologous groups. At genome chromosomes tended to be preferentially eliminated in cells of different ploidy levels.Deletions, insertions, translocations, telocentric chromosomes, isochromosomes and dicentrics and their derivatives were observed in cultured cells. Chromosomes of various homoeologous groups differed in the frequencies and spectra of re—arrangements, but most aberrations occurred in the G-genome chromosomes. In vitro chromosome modifications did not correspond to in vivo variation. Presumably, this difference was caused by differences in the mechanisms of adaptation to the environment at the levels of the cell and the whole organism. G-genome chromosomes were more frequently involved in this process, both in vivo and in vitro.

Notizen: Abstract The hydrogenase of the extremely thermophilic archaebacterium Thermococcus stetteri has been isolated. The procedure involved four steps, the last stage being preparative electrophoresis, with an overall yield of 40%. The molecular mass was estimated to be approx. 106 kDa. Analysis for metal content found iron and nickel in the ratio 13:1. Flavin was also detected in the enzyme and identified as FMN. The hydrogenase activity was relatively insensitive to inhibition by carbon monoxide, Ki 67 μM. This behaviour is indicative of a nickel-iron type hydrogenase. Electron paramagnetic resonance spectroscopy of the enzyme showed signals characteristic of iron-sulfur clusters. The spectra of the hydrogen-reduced T. stetteri hydrogenase, recorded at 70 K, indicated the presence of a single type of [2Fe-2S]cluster. Below 10 K, the spectra were complex, and indicated the presence of other iron-sulfur clusters with extremely fast electron-spin relaxation rates, interpreted as [4Fe-4S] clusters, and minor amounts of [3Fe-4S]clusters. These properties indicate that T. stetteri hydrogenase is related to the hydrogenases of Pyrococcus furiosus, a hyperthermophilic archaebacterium, and Alcaligenes eutrophus.

Notizen: It has been suggested that pyrogenic toxins of Staphylococcus aureus are involved in the series of events leading to some cases of sudden infant death syndrome (SIDS). The objectives of the study were to screen tissues from SIDS infants for pyrogenic toxins and to compare incidence of identification of these toxins among these infants from different countries. An enzyme-linked immunosorbent assay (ELISA) and a flow cytometry method were used to screen body fluids and frozen or formalin-fixed tissues for pyrogenic toxins of S. aureus, toxic shock syndrome toxin 1 (TSST), staphylococcal enterotoxins A (SEA), B (SEB), and C1 (SEC). Toxins were identified in tissues of 33/62 (53%) SIDS infants from three different countries: Scotland (10/19, 56%); France (7/13, 55%); Australia (16/30, 53%). In the Australian series, toxins were identified in only 3/19 (16%) non-SIDS deaths (χ2=5.42, P〈0.02). The flow cytometry method was useful for toxin detection in both frozen and fixed tissues, but ELISA was suitable only for frozen tissues or those fixed for less than 12 months. Identification of pyrogenic toxins in 〉50% of SIDS infants from three different countries indicated further investigation into the role the toxins play in cot deaths might result in development of additional measures to reduce further the incidence of these infant deaths.

Notizen: Abstract Sera obtained from 106 children following an outbreak of Neisseria meningitidis (B:4:P1.15) were screened for bactericidal antibodies against isolates of meningococci and Neisseria lactamica. Most had high titres of antibodies to N. lactamica and N. meningitidis NG:4:- but not to capsulate isolates: B:4:P1.15; B:15:P1.16; B:4:-; C:4:-. Bactericidal activity was higher for both carriers and secretors but the differences were not significant. Bactericidal activity was not associated with total or specific IgA or IgM. Carriers had significantly higher levels of IgG to N. lactamica but not to NG:4:- in sera with bactericidal activity for each of the capsulate strains. Among non-carriers, higher levels of IgG to N. lactamica were associated with killing of B:4:P1.15 and B:4:-. Secretors' sera with bactericidal activity had significantly higher levels of IgG to N. lactamica compared with sera that were not bactericidal. This was not observed among non-secretors. Antibodies to the outbreak strain were adsorbed by all Neisseria isolates tested and absorption of sera with N. lactamica alone completely removed the bactericidal activity against the outbreak strain.

Notizen: This paper presents a real application of a multicriteria decision aid (MCDA) approach to portfolio selection based on preference disaggregation, using ordinal regression and linear programming (UTADIS method; UTilités Additives DIScriminantes). The additive utility functions that are derived through this approach have the extrapolation ability that any new alternative (share) can be easily evaluated and classified into one of several user-predefined groups. The procedure is illustrated with a case study of 98 stocks from the Athens stock exchange, using 15 criteria. The results are encouraging, indicating that the proposed methodology could be used as a tool for the analysis of the portfolio managers' preferences and choices. Furthermore, the comparison with multiple discriminant analysis (either using a stepwise procedure or not) illustrates the superiority of the proposed methodology over a well-known multivariate statistical technique that has been extensively used to study financial decision-making problems.

Notizen: High pressure inactivation kinetics (D and z values) of Saccharomyces cerevisiae ascospores were determined in fruit juices and a model juice buffer at pH 3.5 to 5.0. Approximately 0.5 to 1.0 × 106 ascospores/mL were pressurized at 300 to 500 MPa in juice or buffer. D-values ranged from 8 sec to 10.8 min at 500 and 300 MPa, respectively. The range for z-values was 115 to 121 MPa. No differences (P≥0.05) in D (at constant pressure) or z-values among buffers or juices at any pH were determined, indicating little influence of pH in this range and absence of protective or detrimental effects of juice constituents.

Notizen: We summarize the findings of a two-year study of vegetation and streambank erosion on incised streams. We conducted the first year of the research during the sixth year of a drought. During the second year of study, precipitation totals ranged from normal to 200% of normal. The focus of the study was to determine if vegetation established on a bank affects the erosion of or deposition on that bank. During the drought year, most banks showed relatively little change. During the high water year, 27% of all vegetated and 32% of all bare lower banks retreated more than 250 mm. This similarity between vegetated and unvegetated banks indicates that, on the streams studied, vegetation had little effect on bank erosion. Bank retreat was not related to near-bank velocities or to bank steepness. It is possible that herbaceous vegetation showed no effect on the incised streams because the streams were too far from a new dynamic equilibrium. The energy of the hydraulic system may have been greater than the vegetation could withstand.

Notizen: In trimmed muscle types of four game species the dry matter, crude protein, crude fat, ash, Ca, P, Mg, Na, K, Cu, Zn, Fe, and Mn contents were determined. Samples of the m. semimembranosus and m. longissimus dorsi of each of 10 red deer, fallow deer, roe-deer and wild boars from the Southwestern region of Hungary in the end-winter period were analyzed, and mean values, related to dry matter, were compared. The game muscle samples had much lower crude fat, more Ca, P and microelements, especially Cu and Fe, when compared with published data for the respective muscles of domestic animals.

Notizen: A joint study of microearthquake source and medium parameters was carried out by analysing a seismic sequence that occurred in the Tuscan Emilian Apenninic region. Signal processing and graphic techniques were applied to study the amplitude and polarization of wave motion and thus identify the main and secondary body-wave arrivals on the seismic records.The evidence for similar waveforms (denoted as ‘similar events') in a micro-earthquake subset allowed application of a non-linear technique, which provided the accurate relative location of the events. Microearthquakes occurred within the shallow sediments (average depth of 3.5 km), and all appeared to be concentrated in a small volume of about 1 km3. The composite fault mechanism of the ‘similar events’ was computed by non-linear inversion of P-polarity readings. The maximum-likelihood solution appeared to be well constrained, and indicated a normal-faulting mechanism with the T axis approximately oriented in a northerly direction.Interpretation of several secondary arrivals was performed by direct modelling of traveltimes and wave-motion amplitudes using a double-couple point-source model. Green's functions in a layered medium were computed using two different methods, based on ray theory (Farra 1990) and discrete wavenumber representation of the wavefield (Bouchon 1981). The study of secondary arrivals indicates a depth of 11-12 km for the top of the crystalline pre-Tertiary basement. This estimate concerns a region located at around 3-6 km from the epicentres of the ‘similar events', along the N290°E direction. The modelling of arrival times for an S-to-P converted phase at the basement discontinuity suggests an increase of about 10 per cent in the basement seismic-wave velocity (VP˜˜ 6.9 km s-1), or, equivalently, an increase of the VP to VS ratio in the upper sediments. The increments were defined with respect to an existing reference model by AGIP (Italian Petroleum Agency). Recordings at the Minerbio station (MIN) (located about 40 km SE of the epicentral area) show converted/reflected phases at shallow crustal discontinuities and wave-resonance phenomena. These path effects appear repeatedly on seismic records independently of the size of earthquakes within the magnitude range considered. Waveform and traveltime modelling of these secondary arrivals provide further constraints on the depth and velocity of main-structure discontinuities.

Notizen: Two levels (40 or 80 mg kg−1) of avoparcin were tested as a growth promoter in rainbow trout, Oncorhynchus mykiss (Walbaum), fed for 14 weeks. At 80 mg kg-1 the weight gain was higher (P 〈0.05) than in the unsupplemented control or at the 40 mg kg-1 dose. At either dose, the feed conversion rate and the protein efficiency ratio were more favourable (P 〈0.05) than in the control. Inclusion of the antibiotic significantly improved (P 〈 0.05) the apparent digestibility of lipids. The lipid content of flesh was significantly increased by inclusion of avoparcin in the diet; moreover, at the high level of inclusion the content of some unsaturated fatty acids (18:3ω3 and 20:5ω3) appeared to be significantly increased. No avoparcin residues were detected in the flesh.

Notizen: An analytical model of stream-aquifer interaction is proposed that considers the effects from a small degree of aquifer penetration and low-permeability sediments on the head response to an arbitrary stream-stage hydrograph. Aquifer sections under the stream and beyond are considered in a single model. The model of ground water flow in the aquifer is based on the Dupuit assumptions corrected for leakage from the stream. The model can use stream-stage hydrographs in both analytical and tabular forms. The nondimensional linear boundary value problem is solved for hydraulic head in the aquifer using numerical Laplace transforms and a convolution algorithm. The proposed solution is used to assess the impact of shallow penetration and low-permeability streambed sediments on head responses by comparison with available solutions which neglect these factors.

Notizen: A dipole probe (DP) for measuring vertical variations of hydraulic conductivity has been designed, constructed, and tested in field conditions. The variable-length device designed for the dipole flow test (DFT) consists of three packers separated by changeable spacer rods. The assembled DP is lowered into a well with a long screened interval. The packers are inflated, isolating two screened sections (chambers) between them. A small submersible pump, mounted on the central packer, transfers water from the upper (extraction) chamber into the lower (injection) chamber. This simultaneous injection and extraction creates a recirculatory flow within the aquifer. The chamber head responses are measured by two pressure transducers in the DP chambers. The DP can be used at discrete intervals along the entire well screen length. The DP has been successfully tested in a specially constructed well with a continuous screen in a highly conductive sand and gravel aquifer. During drilling, disturbed samples were collected using the split spoon method and the vertical profile of K was estimated from grain-size analysis. A total of 153 DFTs were performed with various DP geometries. Results indicate that the chamber head changes are sensitive to aquifer properties, reach steady state rapidly, vary linearly with redrculation pumping rate, and can be controlled by changes in DP dimensions. Vertical profiles of K compare well with grain-size analysis estimates.

Notizen: Dimensional analysis is used to show the effects of aquifer anisotropy on capture zone geometry for a partially penetrating well that is screened from the top of a confined aquifer. Dimensionless similarity criteria are identified which allow an extension of results for advective transport modeling from isotropic conditions to anisotropic ones. Using published results of modeling capture zones in isotropic aquifers, it is shown that anisotropy is important in determining the size and shape of the capture zone under two conditions: (1) the pumping rate is small compared to the scaled regional flow velocity, and (2) the well has a low degree of penetration. Examples are used to show that disregard of even modest aquifer anisotropy can significantly affect estimates of capture zone.

Notizen: This paper presents an effective semi analytical method for computation of ground water velocities near a partially penetrating well in an unconfined aquifer. The Neuman solution (1974) is presented as the sum of the Hantush solution (1964) and a correction term for water table conditions. Hydraulic gradients are directly evaluated from the Hantush solution, whereas the gradients for the Neuman correction are calculated using a numerical inversion of the Laplace transform. The method presented here is verified by comparing it with the results of a finite-difference approximation of hydraulic gradients. Calculations of velocities near a pumping well in an unconfined aquifer indicate that, after an initial increase, the vertical velocity decreases before reaching steady state. The method provides a useful tool for the study of well hydraulics and contaminant transport to a pumping well in an unconfined aquifer.

Notizen: 3-Amino- 1,2,4-triazole (amitrole) provided to germinating barley at 20°C in the light led to bleached seedling leaves and photodynamic destruction of chloroplast structure, whereas normal greening and chloroplast ultrastructure was obtained when the seedlings developed in the presence of amitrole in the light at 30°C. Mass spectrometric analysis of the extractable herbicide demonstrated the same content of amitrole in leaves developed at 20 and 30°C. A very similar temperature-sensitive syndrome is characteristic for the nuclear gene mutant ligrina-o34 in barley. Amitrole and the mutation were shown to inhibit the cyclization of lycopene, leading to severe deficiencies in β-carotene and its xanthophyll derivative lutein. Besides accumulation of lycopene, also its precursors phytoene, phytofluene and ξ-carotene accumulated. Inhibition of carotenoid biosynthesis by amitrole and the mutation at 20°C in the light led to a strong reduction of both transcript and protein levels for chloroplastic glutamine synthetase (GS2) while transcript amount and protein of the cytosolic isoenzyme (GS1) were unaffected. At 30°C increased levels of mRNA for the chloroplastic isoform GS2 were observed in wild type, mutant and amitrole-treated seedlings, but protein levels remained unchanged. Turnover rates of the GS2 protein were the same at 20 and 30°C. This extensive translational control of chloroplastic GS2 synthesis was also observed in a heat shock experiment, which revealed transiently increased mRNA levels for chloroplastic GS2 but unchanged protein levels.Permissive synthesis of β-carotene and chloroplastic glutamine synthetase (GS2) at 30°C in the presence of amitrole or the tigrina-o34 mutation might be due to two alternative pathways of ionone ring formation using either lycopene or neurosporene as substrates for cyclization.

Notizen: Bacterial cell surface hydrophobicity is one of the most important factors that influence bacterial adhesion. A new method, microsphere adhesion to cells, for measuring bacterial cell surface hydrophobicity was developed. Microsphere adhesion to cells is based on microscopic enumeration of hydrophobic, fluorescent microspheres attaching to the bacterial surface. Cell surface hydrophobicity estimated by microsphere adhesion to cells correlates well with adhesion of bacteria to hydrocarbons or hydrophobic interaction chromatography for a set of hydrophilic and hydrophobic bacteria (linear correlation coefficients, R2, were 0.845 and 0.981 respectively). We also used microsphere adhesion to cells to investigate the in situ properties of individual free-living bacteria directly in activated sludge. Results showed that the majority of the bacteria were hydrophilic, indicating the importance of cell surface hydrophobicity for bacterial adhesion in sludge, and for the overall success of the wastewater treatment process.

Notizen: Although leaf photosynthesis and plant growth are initially stimulated by elevated CO2 concentrations, increasing insensitivity to CO2 (acclimation) is a frequent occurrence. In order to examine the acclimation process, we studied photosynthesis and whole plant development in swiss chard (Beta vulgaris L. Koch ssp. ciela) and sugarbeet (Beta vulgaris L. ssp. vulgaris) grown at either ambient or twice ambient concentrations of CO2. In an initial controlled environment study, photosynthetic acclimation to elevated CO2 levels was observed in both subspecies 24 days after sowing (DAS) but was not observed at 42 and 49 DAS for sugarbeet or at 49 DAS for swiss chard. Although sugarbeet and swiss chard differed in root size and morphology, this was not a factor in the onset of photosynthetic acclimation. The reversal of photosynthetic acclimation that was observed in older plants grown at elevated CO2, concentrations was associated with a rapid increase in root development (i.e. increased root: shoot [R/S] ratio), increased sucrose levels in sinks (roots) and no differences in total soluble leaf protein of either subspecies relative to the ambient CO2 condition. In a second set of experiments, swiss chard and sugarbeet were grown in outdoor Plexiglass chambers at different times of the year (i.e. summer and early fall). Average 24-h temperature was 30.7 and 19.4°C for the summer and fall plantings, respectively. In agreement with the controlled environment study, lack of photosynthetic acclimation, determined from the response of photosynthesic rate to internal CO2 concentration, was correlated with increased root biomass and sucrose concentration relative to the ambient condition. However, photo-synthetic acclimation was observed depending on the season, i.e. summer (swiss chard) or fall (sugarbeet), suggesting that acclimation was affected by environmental factors, such as temperature. Data from both experiments suggest that continued long-term photosynthetic stimulation may be dependent upon the ability of increased CO2 to stimulate new sink development which would allow full utilization of the additional carbon made available in a high CO2 environment.

Notizen: Three soybean (Glycine max L. Merr.) cultivars (Maple Glen, Clark and CNS) were exposed to three CO2 concentrations (370, 555 and 740 μmol mol−1) and three growth temperatures (20/15°, 25/20° and 31/26°C, day/night) to determine intraspecific differences in single leaf/whole plant photosynthesis, growth and partitioning, phenology and final biomass. Based on known carboxylation kinetics, a synergistic effect between temperature and CO2 on growth and photosynthesis was predicted since elevated CO2 increases photosynthesis by reducing photorespiration and photorespiration increases with temperature. Increasing CO2 concentrations resulted in a stimulation of single leaf photosynthesis for 40–60 days after emergence (DAE) at 20/15°C in all cultivars and for Maple Glen and CNS at all temperatures. For Clark, however, the onset of flowering at warmer temperatures coincided with the loss of stimulation in single leaf photosynthesis at elevated CO2 concentrations. Despite the season-long stimulation of single leaf photosynthesis, elevated CO2 concentrations did not increase whole plant photosynthesis except at the highest growth temperature in Maple Glen and CNS, and there was no synergistic effect on final biomass. Instead, the stimulatory effect of CO2 on growth was delayed by higher temperatures. Data from this experiment suggest that: (1) intraspecific variation could be used to select for optimum soybean cultivars with future climate change; and (2) the relationship between temperature and CO2 concentration may be expressed differently at the leaf and whole plant levels and may not solely reflect known changes in carboxylation kinetics.

Notizen: Soybean (Glycine max cv. Clark) was grown at both ambient (ca 350 μmol mol−1) and elevated (ca 700 μmol mol−1) CO2 concentration at 5 growth temperatures (constant day/night temperatures of 20, 25, 30, 35 and 40°C) for 17–22 days after sowing to determine the interaction between temperature and CO2 concentration on photosynthesis (measured as A, the rate of CO2 assimilation per unit leaf area) at both the single leaf and whole plant level. Single leaves of soybean demonstrated increasingly greater stimulation of A at elevated CO2 as temperature increased from 25 to 35°C (i.e. optimal growth rates). At 40°C, primary leaves failed to develop and plants eventually died. In contrast, for both whole plant A and total biomass production, increasing temperature resulted in less stimulation by elevated CO2 concentration. For whole plants, increased CO2 stimulated leaf area more as growth temperature increased. Differences between the response of A to elevated CO2 for single leaves and whole plants may be related to increased self-shading experienced by whole plants at elevated CO2 as temperature increased. Results from the present study suggest that self-shading could limit the response of CO2 assimilation rate and the growth response of soybean plants if temperature and CO2 increase concurrently, and illustrate that light may be an important consideration in predicting the relative stimulation of photosynthesis by elevated CO2 at the whole plant level.

Notizen: The aerobic biodegradation of racemic mixtures of five chiral phenoxyalkanoic acids was studied according to a biodegradation test that was complemented with enantiomer-specific analysis. Both enantiomers of (RS)-2-phenoxypropanoic acid, (RS)-2-(3-chlorophenoxy)propanoic acid, and (RS)-2-(4-chlorophenoxy)propanoic acid, were completely degraded within 25 days when aerobically incubated with activated sludge. During incubations of (RS)-2-phenoxypropanoic acid, the (R) enantiomer was degraded before the (S) enantiomer, whereas during incubations of (RS)-2-(3-chlorophenoxy)propanoic acid the (S) enantiomer was preferentially degraded. The (R) enantiomer of (RS)-2-(2-chlorophenoxy)propanoic acid was removed after 24 days while only 30% of the (S) enantiomer was degraded within 47 days of incubation. (RS)-2-(2,4,5-Trichlorophenoxy)propanoic acid was the most persistent of all the racemic mixtures tested. After 47 days, the concentration of the (S) enantiomer was nearly unchanged, and the concentration of (R)-2-(2,4,5-trichlorophenoxy)propanoic acid had decreased only by about 40%. The differences observed in the length of the lag phases and in the degradation rates of individual enantiomers can lead to accumulations of the more recalcitrant enantiomer in aquatic or terrestrial ecosystems.

Notizen: This paper examines the efficacy of currency swaps as a hedging mechanism for the exchange rate risk associated with foreign investment in real estate. Earlier studies have concentrated on short-term hedging instruments such as options and forward contracts. Currency swaps are better suited for use on investments with long-term holding periods such as real estate. The findings indicate that, although hedging United States real estate investments with currency swaps suppresses most of the risk induced by currency instability, the improvements are insufficient to produce diversification gains for foreign investors in the context of mean-variance portfolio performance.

Notizen: Liang, Myer and Webb (1996) have offered bootstrap simulation as a tool for quantifying the uncertainty in the optimum composition of portfolios. Unfortunately, the confidence intervals produced were so large, they were unable to provide any new insight to the question, “How Much in Real Estate?”. In this comment, adjustments have been made to the methodology they proposed and as a result have produced findings which lead to very different conclusions. More specifically, the results suggest that investors with a low risk preference should hold very little real estate.

Notizen: HIV encephalitis is the common pathologic correlate of HIV-dementia (HAD). HIV-infected brain mononuclear phagocytes (MP) (macrophages and microglia) are reservoirs for persistent viral infection. When activated, MP contribute to neuronal damage. Such activated and virus-infected macrophages secrete cellular and viral factors, triggering neural destructive immune responses. Our Center's laboratories have begun to decipher the molecular and biochemical pathways for MP-mediated neuronal damage in HAD. This review will discuss the salient clinical and pathological features of HAD and highlight the recent advances made, by our scientists and elsewhere, in unraveling disease mechanisms, including the role of chemokines and their receptors in the neuropathogenesis of HIV-1 encephalitis.

Notizen: The effect of a pipeline corridor constructed through an ecological reserve in Southern California was investigated by assessing plant species composition and soil chemistry. A homogeneous plant community comprised primarily of exotic annuals was found along the entire length of the corridor. This community has low similarity to the adjacent native plant communities. Soil organic matter was significantly less on the disturbed corridor than in contiguous undisturbed areas. Both available nitrogen and extractable phosphorus values were greater in the disturbed corridor. By contrast, total nitrogen was significantly higher outside the pipeline. The more labile litter of the exotic annuals allows increased mineralization along the corridor than does the more recalcitrant litter of the native perennial shrubs in the undisturbed areas. Once established, the weedy exotic annual litter may completely turn over organic matter and nitrogen, favoring the persistence of the weedy annuals. These exotic annuals appear to be moving into three of the native communities - grassland, coastal sage, and oak woodland - that have less organic matter and a more open plant canopy. Poor restoration efforts can lead to the establishment of such exotics, subsequent invasion into the surrounding undisturbed habitat, and degradation of the reserve.

Notizen: Abstract End-member, continuous and degenerate reactions are derived for the multisystem with the six components Na2O, CaO, (Mg/Fe)O, Al2O3, SiO2, H2O among the phases plagioclasess, garnetss, amphiboless, cpx, opx, olivine, spinel, quartz and an aqueous fluid. The chemography of this system is degenerate due to the co-linearity 2Opx = Ol + Qtz. This co-linearity has its implications both on reaction space and phase equilibria.From a total of 28 reaction systems, reaction space is derived for nine subsystems (phases in parentheses are absent): Case A1: (Cpx,Ol) (Cpx,Opx) and (Cpx,Qtz), Case A2: (Spl,Ol) (Spl,Opx) and (Spl,Qtz), Case B: (Ol,Opx) (Ol,Qtz) and (Opx,Qtz).In the absence of either cpx or spl (case A), three reactions form an invariant point, either [Cpx] or [Spl], where the co-linear phases olivine, opx and quartz coexist on the transformation line 2Opx = Ol + Qtz. Changing mineral compositions force invariant points to move along the line with the different reaction curves changing their relative position according to Schreinemakers’rules. Zero contours, i.e. the location where (a) phase(s) disappear(s) in reaction space correspond to singular points in phase diagrams. Two types are distinguished; singular points of indispensable and of substitutable phases. In the first case the phase disappears from the entire bundle while in the second it disappears from a single reaction. In the specific case where the substitutable phases are also the co-linear ones, two of the three co-linear phases disappear simultaneously. Two of the three reaction curves coincide.In the system including Cpx and Spl (Case B) three reactions, (Ol,Opx) (Ol,Qtz) and (Opx,Qtz), oppose three invariant points, [Ol], [Opx] and [Qtz]. Invariant points no longer move along the line 2Opx = Ol + Qtz. The coincidence of the zero contours of all three co-linear phases in reaction space-the result of the chemographic degeneracy-causes the respective singular points to coincide in the phase diagrams. This is the location where curves must be rearranged in a bundle to conform Schreinemakers’rules.The reaction Grs1Prp2= 2 Ol + An is fourth order degenerate and part of all nine subsystems (cases A and B). It can be used to relate the different phase diagrams to one another.

Notizen: The Rhodobacter sphaeroides mutants Drn12 and Drn21 derepressed for nitrogenase synthesis in the presence of ammonia and impaired in utilization of certain nitrogen sources have been analyzed. Both mutants show a low level of expression of the glnBA operon. The DNA fragment restoring the wild-type phenotype to these mutants contains the 3′-portion of ntrB gene and the entire ntrC gene. Sequence analysis showed that Drn12 bears a missense mutation in the ntrC gene. The mutation results in the replacement of a glycine residue by aspartate within the N-terminal domain of the NtrC protein. Pleiotropic phenotypes of Drn12 and Drn21 appear to be associated with an alteration in the regulation of glnBA expression.

Notizen: Soils from seven sites on the island of Jamaica were assayed for the symbiotic diazotrophs Frankia and Rhizobium using serial dilutions. Most probable number and least squares regression methods were used to estimate each soil's capacity to nodulate native Myrica cerifera, exotic Leucaena leucocephala and exotic Casuarina cunninghamiana. The sample sites included a montane forest, a slash-and-burn agricultural site, reclaimed bauxite mining areas, abandoned sugar cane fields, and a garden plot. None of the host plants used in the bioassay were present on the sites sampled except for scattered L. leucocephala on one site. Frankia capable of nodulating M. cerifera, which is native to Jamaican highlands, occurred at all sites sampled. No C. cunninghamiana-infective Frankia was detected in soils sampled. Only soils from one site on the tropical coastal plain harbored rhizobia able to nodulate L. leucocephala (37 nodulation units cm−3 of soil). A subset of nodulated M. cerifera and L. leucocephala reduced acetylene to ethylene indicating nitrogenase activity. The slash-and-burn agricultural site, which was situated at an elevation of 200 m and possessed both high natural fertility and high soil moisture-supplying capacity, had significantly greater Myrica infectious capacity (1 000 nodulation units cm−3 of soil) than the other sites (7−207 nodulation units cm−3 of soil). A planned, paired comparison revealed that a recently cultivated sugar cane field and a recently reclaimed bauxite mining site together had significantly less Myrica-infective Frankia (4 nodulation units cm−3 of soil) than a corresponding pair of sites consisting of a sugar cane field abandoned for 25 years and a bauxite mining site reclaimed 20 years before sampling (118 nodulation units cm−3 of soil). Results indicate that Myrica-infective Frankia is widespread in Jamaica, that the number of Myrica-infective Frankia units vary from site to site in accordance with soil type and soil history, that Jamaican sites sampled lack soil Frankia populations capable of nodulating a casuarina host, that rhizobial symbionts capable of nodulating L. leucocephala may be geographically restricted to lowlands in Jamaica, and that the occurrence of Frankia in these soils is independent of host plant presence.

Notizen: We examined the effectiveness of Dual Attribute Time Intensity (DATI) method for assessment of temporal changes in perceived toughness and juiciness, within commercially acceptable meat cuts. Usefulness of DATI in assessing temporal aspects of perception of juiciness and toughness was compared with Single-Attribute Time-Intensity (SATI) and Line Scale Profile. Results showed that DATI provided a good separation of attributes and was equal to or better than SATI in differentiating beef samples based on perceived juiciness and toughness. By reducing the dumping effect and the inherent sample to sample variability, this method enabled more precise assessment of the relationship between juiciness and toughness in meat than SATI.

Notizen: The degradation kinetics of (−)-epigallocatechin gallate (EGCG) as a function of pH (4 to 7) and dissolved oxygen concentration (0 to 7.59 mg/L) in citrate buffer solutions were determined. EGCG degradation followed a pseudofirst order kinetic model, regardless of the conditions when the fractional conversion technique was applied for data reductions. The rate constant increased log-linearly with respect to both pH and dissolved oxygen concentration. The temperature dependence of the rate constant was adequately modeled by the Arrhenius equation with an average activation energy of 18.7 ± 1 kcal/mol. A mathematical model that relates the rate constant to temperature, pH, and dissolved oxygen concentration was developed and validated. These results can help predict losses of EGCG during processing and/or storage.

Notizen: Abstract: We review and update the work on genetic elements, e.g., viruses and plasmids (excluding IS elements and transposons) in the kingdom Crenarchaeota (Thermoproteales and Sulfolobales) and the orders Thermococcales and Thermoplasmales in the kingdom Euryachaeota of the archael domain, including unpublished data from our laboratory. The viruses of Crenarchaeota represent four novel virus families. The Fuselloviridae represented by SSV1 of S. shibatae and relatives in other Sulfolobus strains have the form of a failed spindle. The envelope is highly hydrophobic. The DNA is double-stranded and circular. Members of this group have also been found in Methanococcus and Haloarcula. The Lipothrixviridae (e.g., T TV1 to 3) have the form of flexible filaments. They have a core containing linear double-stranded DNA and DNA-binding proteins which is wrapped into a lipid membrane. The ‘Bacilloviridae’ (e.g., TTV4 and SIRV) are stiff rods lacking this membrane, but also featuring linear double-stranded DNA and DNA-binding proteins. Both virus type carry on both ends structures involved in the attachment to receptors. Both types are represented in Thermoproteus and Sulfolobus. The droplet-formed novel Sulfolobus virus SNDV represents the ‘Guttaviridae’ containing circular double-stranded DNA. Though head and tail viruses distantly resembling T phages or lambdoid phages were seen electronmicroscopically in solfataric water samples, no such virus has so far been isolated. SSV1 is temperate, TTV1 causes lysis after induction, the other viruses found so far exist in carrier states. The hosts of all but TTV1 survive virus production. We discuss the implications of the nature of these viruses for understanding virus evolution. The plasmids found so far range in size from 4.5 kb to about 40 kb. Most of them occur in high copy number, probably due to the way of their detection. Most are cryptic, pNOB8 is conjugative, the widespread pDL10 alleviates in an unknown way autotrophic growth of its host Desulfurolobus by sulfur reduction. The plasmid pTIK4 appears to encode a killer function. pNOB8 has been used as a vector for the transfer of the lac S (β-galactosidase) gene into a mutant of S. solfataricus.

Notizen: PNPase and RNase II are the key regulatory exo-nucleases controlling mRNA decay in Escherichia coli The rnb transcripts were found to proceed through the terminator and PNPase was found to be involved in the 3’to 5’degradation of rnb mRNA. Analysis of these longer 3’termini revealed that they are located in UA-rich regions. Comparison of single and double mutants suggested that PNPase and RNase II could have different roles in the degradation of these unstructured regions. We have shown that RNase II levels can vary over a fivefold range in haploid cells and that its expression depends on PNPase levels. PNPase-deficient strains were found to have a 2–2.5-fold increase in RNase M activity, while PNPase-overproducing strains reduced the rnb message and RNase II levels. Conversely, the amount of PNPase in the rnb deletion strain was approximately twofold higher than that in the wild-type strain. These observations suggest that the two main exonu-cleases are inter-regulated through a fine tuning mechanism. We discuss the implications of these results with regard to mRNA degradation and cell metabolism.

Notizen: Abstract Ribonuclease II (RNase II), encoded by the rnb gene, is one of the two major Escherichia coli exonucleases involved in mRNA degradation. Some of the ribonucleases implicated in this process have recently been shown to be inter-regulated. In this paper we studied the effects of the endonucleases RNase E and RNase III in rnb expression. We have shown that RNase E cleaves the rnb message internally: when this ribonuclease is inactivated rnb mRNA accumulates with a concomitant increase in RNase II activity. RNase III also affects RNase II expression but in an indirect way. We discuss these implications for the regulation of mRNA degradation.

Notizen: A method for monitoring water flow and solute transport through the unsaturated zone is of crucial necessity for early detection of threats to ground water. A method developed by Ukrainian hydrogeologists in the 1970s involves a borehole of a large enough diameter for a person to enter, that provides the possibility of installing monitoring equipment, and enables collecting undisturbed sediment blocks and samples of pore water and gas. Practically, tensiometers and vacuum lysimeters are installed at various levels in the borehole walls. They provide high quality sampling and measurements, and enable determining parameters of unsaturated water flow and solute transport and their temporal and spatial variability.The method was tested at several locations. Selected results from two sites, showing the temporal and spatial distributions of capillary pressure and solute concentrations, are presented. The results exhibit the difference between lateral variation in water flow and solute transport characteristics. The accuracy and representativeness of the measurements were tested against the mutual interaction between the various instruments installed in the borehole walls. Monitoring using the large-diameter boreholes provides the possibility of early detection of threats to ground water under areal contamination inputs, and of quantitative prediction of its future trends.

Notizen: : The aim of this paper is to present a measure of regional labour costs in manufacturing industry in the European Community in 1988. As raw data, derived from the Eurostat Labour Cost Survey, are strongly dependent on sectoral distribution of employment in each region, a method is presented, based on reweighting techniques and loglinear modelling, to eliminate the composition effect. As a result, a regular geographical pattern does emerge. The same type of technique is then applied to the evolution of regional labour costs between 1984 and 1988; some correlation does appear with regional unemployment levels in the United Kingdom and France, pointing the way towards further research.

Notizen: In 1993, the Research School of Earth Sciences of the Australian National University commenced a nationwide seismometry project, SKIPPY, for a comprehensive study of the upper mantle beneath the Australian continent. We applied a waveform inversion technique to broad-band data recorded while the SKIPPY portable arrays were positioned in eastern Australia in order to construct a 3-D model of shear velocity in the upper mantle and transition zone beneath eastern Australia and the adjacent oceanic regions. The SKIPPY data were augmented by data from the permanent seismological observatories in the region. The first step of the waveform inversion used involved the matching of the waveforms of fundamental- and higher-mode Rayleigh waves with waveforms synthesized from radially stratified models; in the second stage the linear constraints on radial variations in shear velocity were combined in a tomographic inversion for aspherical variations in shear velocity. The preferred model reduces the data variance by 90 per cent. Owing to the dense data coverage, structural features with dimensions larger than 250 km laterally and 50 km vertically are resolved. The major structural features inferred from the tomographic images are (1) a substantial increase in the thickness of the high-velocity ‘lid’ from the oceanic region to the Phanerozoic continental and Proterozoic continental regions, (2) a pronounced low-velocity zone centred at 140 km depth beneath the part of eastern Australia often referred to as the Tasman fold belt, which confirms previous inferences from Rayleigh-wave dispersion curves, (3) a prominent zone of low wave speeds beneath the eastern margin of the continent that coincides with locations of Cenozoic volcanism and regions of enhanced heat flow, (4) deep low-wave-speed anomalies beneath both the Coral and Tasman seas, and (5) localized high-wave-speed perturbations in the mantle transition zone (410-660 km depth) beneath eastern Australia, in particular beneath the New England fold belt and the Mount Isa block. From the images we infer that the eastern boundary of the Precambrian shields does not coincide with a sharp seismic contrast, unless this boundary is located further to the east than is commonly assumed. The structure of the upper mantle and transition zone beneath eastern Australia as inferred from the images is more complex than that beneath the Precambrian cratons of central and western Australia, which could be due to subduction beneath the eastern continental margin prior to opening of the Tasman sea. If correct, this interpretation would imply that part of the upper mantle has moved along laterally with the lithosphere during the relatively fast northward motion of the plane.

Notizen: Plasmodium gallinaceum ookinetes adhered to Aedes aegypti midgut epithelia when purified ookinetes and isolated midguts were combined in vitro. Ookinetes preferentially bound to the microvillated luminal surface of the midgut, and they seemed to interact with three types of structures on the midgut surface. First, they adhered lo and migrated through a network-like matrix, which we have termed microvilli-associated network, that covers the surface of the microvilli. This network forms on the luminal midgut surface in response to blood or protein meals. Second, the ookinetes bound directly to the microvilli on the surface of the midgut and were occasionally found immersed in the thick microvillar layer. Third, the ookinetes associated with accumulations of vesicular structures found interspersed between the microvillated cells of the midgut. The origin of these vesicular structures is unknown, but they correlated with the surface of midgut cells invaded by ookinetes as observed by TEM. After binding to the midgut. ookinetes underwent extensive morphological changes: they frequently developed one or more annular constrictions, and their surface roughened considerably, suggesting that midgut components remain bound to the parasite surface. Our observations suggest that, in a natural infection, the ookinete interacts in a sequential manner with specific components of the midgut surface. Initial binding to the midgut surface may activate the ookinete and cause morphological changes in preparation for invasion of the midgut cells.

Notizen: Abstract: Replication of satellite phage P4 of Escherichia coli is dependent on three phage-encoded elements: the origin (ori), a cis replication element (crr), and the product of the α gene, gpα. In vitro P4 replication is origin-specific resulting in monomeric form I DNA. DNA synthesis requires chromosomally encoded proteins DNA polymerase III holoenzyme, SSB, DNA gyrase and probably topoisomerase I; host-encoded initiation and priming functions are dispensable. The α protein is multifunctional in P4 replication, combining three activities in a single polypeptide chain. First, the protein complexes specifically with type I repeats at ori and crr. Second, the helicase activity associated with gpα unwinds DNA with 3′→ 5′ polarity. Third, the primase activity results in the synthesis of RNA primers. Defined sequence motifs in gpα correlate with the helicase and primase activities which are arranged in distinct, separable domains. Primase activity is associated with the N-terminal half of the protein, ori / crr binding with the C-terminal portion. A model for the initiation mechanism of P4 replication which resembles that of mammalian simian virus 40 is discussed.

Notizen: As the world economy begins to shift from the industrial states of the past toward marketable service orientations, the need for those adults under highly stressful circumstances to reduce their stress levels becomes imperative. The peak performer, whether in business and industry, education and teaching or any other field, should regard, accept and anticipate stress indicators by using appropriate methods to alleviate that stress before it becomes highly detrimental to employment and personal performance. The utilization of those activities we played as children can become modified tools and techniques for the stressed personality.

Notizen: A method for finding all deterministic and maximally general rules for a target classification is explained in detail and illustrated with examples. Maximally general rules are rules with minimal numbers of conditions. The method has been developed within the context of the rough sets model and is based on the concepts of a decision matrix and a decision function. The problem of finding all the rules is reduced to the problem of computing prime implicants of a group of associated Boolean expressions. The method is particularly applicable to identifying all potentially interesting deterministic rules in a knowledge discovery system but can also be used to produce possible rules or nondeterministic rules with decision probabilities, by adapting the method to the definitions of the variable precision rough sets model.

Notizen: Three radiocarbon dates on marine organic material from southernmost Spitsbergen are presented, making it possible to compare the Holocene shoreline displacement in this area with that of other areas of Svalbard. The most distinct raised beach feature is the large beach ridge terrace about 10 m above sea level which most probably is the result of a marine transgression ca. 6500 years ago. Dating of 2700-year-old laminaria from about 2 m above sea level demonstrates a small rising of the land during the youngest Holocene.

Notizen: Abstract. Ammonia volatilization with and without gypsum incorporation was measured in Gujranwala soil (Udic Haplustalf) in an incubation study using different nitrogen fertilizers e.g. urea, ammonium sulphate (AS), calcium ammonium nitrate (CAN), and urea nitrophos (UNP). Nitrogen from different fertilizers was applied at the rate of 200 mg N kg−1 to two sets of soils in plastic bags (1.0 kg soil) and plastic jars (0.5 kg soil). Soil moisture was maintained at field capacity. Application of urea increased soil pH to 9, three hours after its addition. Ammonium sulphate and calcium ammonium nitrate had little effect on soil pH. Ammonium volatilization losses from fertilizers were related to the increase in soil pH caused by the fertilizers. Consequently maximum losses were recorded due to application of urea. Losses through ammonia volatilization were significantly lower with AS, CAN and UNP in descending order. Gypsum incorporation significantly reduced the losses. Therefore, application of gypsum to soil before urea may substantially improve N use efficiency for crop production by reducing N losses.

Notizen: The effect of phenylacetic acid (PAA) on rice (Oryza saliva L.) anther culture was investigated with six genotypes, using 2,4-D as control. In the two-step culture protocol, replacing 2, 4-D with PAA in the induction medium did not influence callus induction but significantly improved the shoot differentiation from callus, particularly in the indica cultivar Teqing. The anther-derived calli of all genotypes regenerated shoots directly on the callus induction medium containing PAA. Most of the directly-regenerated plantlets had well-developed root systems and were therefore readily transplanted into soil. The improved shoot differentiation potential and the frequency of direct regeneration depended on genotype, basal medium and PAA concentration. The one-step green shoot regeneration frequencies obtained were 1.98% with the indica cultivar ‘129’, 1.5% with the indica × japonica hybrid ‘Teqing/02428’ (F1), and 1.98% with the indica × indica hybrid ‘Waiyin 2/C.B.’ (F1). The PAA-based one-step method was most effective on the anther culture of indica genotypes. Three DH populations have been constructed from hybrids (F1) via one-step culture. PAA also enhanced the one-step plantlet formation in rice somatic tissue culture.

Notizen: The effects of sodium acetate, sodium lactate, and propyl gallate were determined on microbial populations, pH, ammonia content and color of film-overwrapped or vacuum-skin packaged fresh shrimp and catfish fillets. Products were stored at 4°C up to 12 days. Vacuum-skin packaged shrimp or catfish fillets contained lower (P 〈 0.05) ammonia compared to film-overwrapped products. Catfish fillets treated with 2% sodium acetate had lower populations of psychrotrophic and anaerobic bacteria compared with untreated catfish, regardless of packaging. Treatment with 2% NaAc has potential for extending shelf life of refrigerated catfish fillets.

Notizen: Boshan is an industrial city in the center of Shandong Province where ground water is the only source for the urban water supply. The major water resource is fracture-karst water in the middle Ordovician carbonate rocks. Based on the hydrogeological investigation and mapping in this area we studied the geologic and hydrogeologic settings, the major pollution sources and the pathways of contamination, the principal contaminants, and their spatial distribution in ground water. The ground-water quality has also been estimated by the fuzzy mathematic method. The geostatistical method, such as the kriging method, was taken to simulate spatial distribution of the contaminants. The grey system method was adopted to forecast future contamination. An attempt at the remediation of Cr6+ contamination in fracture-karst water was also discussed. Finally, some proposals for the protection of the ground-water environment in Boshan District are offered.

Notizen: Abstract Various residual stress conditions were introduced into the surface of a 7075-T6 aluminium alloy by shot peening. This resulted in a considerable alteration in subsequent fatigue crack growth behaviour. A systematic investigation into the effects of a variety of conditions on fatigue crack propagation behavior was conducted. The degree of shot peening was established using the Almen scale and calibration strips. Changes occurred in crack re-initiation lives, crack growth rate and hardness across the thickness profile. Residual stresses were introduced by various peening operations (such as different peening positions and loading specimens prior to and during peening). By measuring the crack opening level of peened specimens, changes in fatigue crack closure effect with respect to the crack extension in CT specimens are reported. The relations of re-initiation life versus crack opening level and Almen levels, and of the crack closure effect versus Almen scales, were addressed experimentally. It was found that the improvement in re-initiation life and the retarding effect of crack growth, both influenced by the Almen scales, are associated directly with the elevation of crack opening level, particularly at the stage of crack re-initiation.

Notizen: Paired A- and B-lines, cytoplasmic male-sterile (CMS)-lines and main-tainer lines, respectively, were crossed with R-lines (restorer) to produce A × R, R × B and B × R hybrids, which were used in 1993 and 1996 to predict the performance of three lypes of CMS cotton lines available in China. A significant difference in yield and yield components was revealed between paired A × R and B × R hybrids. This difference was greatly influenced by both CMS cytoplasm and the interaction between cytoplasm and nuclear genotypes. It is suggested that there are detrimental effects of CMS cytoplasm on yield and yield components. General combining ability of near-isogenic CMS lines was also affected by this negative effect. The detrimental effect was closely related to an increased number of immature seeds per boll, which might be caused by partial female sterility associated with CMS cytoplasm. The possibility of developing specific combinations of CMS Upland × Upland restorer hybrids that express enough heterosis for yield to overcome the detrimental effect of the CMS cytoplasm is discussed.

Notizen: Fresh pale, soft, exudative (PSE), dark, firm, dry (DFD), and normal pork were stored under light or dark conditions at 4°C for 7 days. Sample pH, metmyoglobin reductase activity, oxygen consumption rate, and relative surface metmyoglobin and oxymyoglobin contents were determined. DFD pork had the highest metmyoglobin reductase activity and oxygen consumption rate. Enzyme activity of PSE was lower than that of normal pork, but no difference existed in oxygen consumption rate between PSE and normal samples. Metmyoglobin reductase activity dropped slowly during meat storage; oxygen consumption rate sharply decreased during the first day of storage. Both metmyoglobin reductase activity and oxygen consumption rate declined more rapidly in the light. Results can help develop guidelines for display and packaging of pork.

Notizen: Pale, soft, exudative (PSE), normal, and dark, firm, dry (DFD) pork was held 7 days at 4°C under light or dark then color was evaluated instrumentally and visually. Change in reflectance (R630-R580) was used to estimate color changes due to oxymyoglobin. PSE pork was lighter (high L*) than normal and DFD pork, but did not change over time. An increase in a* and R630-R580 occurred the first day for normal and DFD, but not for PSE samples. A subsequent increase in hue angle and decrease in R630-R580 (loss of redness) occurred in PSE and normal, but not in DFD samples. Changes in hue angle and R630-R580 were more rapid in PSE than in normal samples. Light exposure accelerated all changes except L*. Visual redness inversely correlated with L* value.

Notizen: H6 is an Atriplex nummularia gene having high sequence homology with the algal caltractin (a basal-body-associated calcium-binding protein) gene. Recombinant H6 was expressed at high levels in Escherichia coli. The recombinant protein exhibited an ethylene glycol-bis(β-aminoethyl ether)-N,N,N′,N′-tetraacetic acid-induced mobility shift during sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and it was able to bind 45Ca2-. An H6 cDNA probe detected three RNA transcripts of 1.3, 2.2 and 2.4 kb, respectively. The levels of these transcripts were regulated by different environmental cues and during developmental stages. The steady state levels of the 1.3-kb mRNA decreased after touch and heat treatments. Expression of the 2.2-kb message correlated with cell proliferation activity. In cultured cells, the highest level of the 2.2-kb message preceeded the peak of mitotic cell division activity. In plants, the 2.2-kb message was detected only in shoot tips that contained meristematic tissues. The 2.4-kb message was detected exclusively in heat-shocked cells. The relationship among the three transcripts is discussed in the context of the possible role of H6 in mediating developmental and environmental signals.

Notizen: The absolute pressure in conducting xylem vessels of roots of 2-week-old, slowly transpiring intact maize plants (bathed in nutrition medium) was determined to be +0·024 ± 0·044 MPa using the xylem pressure probe. When the roots were subjected to osmotic stress (NaCI, KCI or sucrose), the xylem pressure decreased immediately and became more negative. However, the response of xylem pressure to osmotic stress was considerably attenuated, indicating that the radial reflection coefficients, σ13 of the maize root for these solutes were rather low (between 0·2 and 0·4 depending on the concentration of the osmoticum). The low values of a, may be caused (partly) by unstirred layer effects. In repeated osmoticum/nutrition regimes a complex pattern of changes in xylem pressure was observed which was apparently linked to the interplay between transpiration and (passive and/or active) solute loading of the xylem. These processes were not observed when the roots were subjected to osmotic stress after excision. In this case, a biphasic response was observed comparable to that found for excised roots using the root pressure probe.

Notizen: Replication protein A (RPA) consisting of three subunits is a eukaryotic single-stranded DNA (ssDNA)-binding protein involved in DNA replication, repair and recombination. We report here the identification and characterization of a RPA large subunit (CpRPA1) gene from the apicomplexan Cryptosporidium parvum. The CpRPA1 gene encodes a 53.9-kDa peptide that is remarkably smaller than that from other eukaryotes (i.e. ∼70 kDa) and is actively expressed in both free sporozoites and parasite intracellular stages. This short-type RPA large subunit has also been characterized from one other protist, Crithidia fasciculata. Three distinct domains have been identified in the RPA large subunit of humans and yeasts: an N-terminal protein interaction domain, a central ssDNA-binding area, and a C-terminal subunit-interacting region. Sequence analysis reveals that the short-type RPA large subunit differs from that of other eukaryotes in that only the domains required for ssDNA binding and heterotrimer formation are present. It lacks the N-terminal domain necessary for the binding of proteins mainly involved in DNA repair and recombination. This major structural difference suggests that the mechanism for DNA repair and recombination in some protists differs from that of other eukaryotes. Since replication proteins play an essential role in the cell cycle, the fact that RPA proteins of C. parvum differ from those of its host suggests that RPA be explored as a potential chemotherapeutic target for controlling cryptosporidiosis and/or diseases caused by other apicomplexans.