Patterns of Root Colonization in Epacridaceous Plants Collected from Different Sites

Root colonization was studied in ten species of the Epacridaceaeat three sites in Victoria by morphological and cross-inoculationexperiments. The sites and genera chosen were Cranbourne [Epacrisimpressa Labill. andLeucopogon ericoides(Smith) R. Br.] andRye [L. parviflorus(Andrews) Lindley] on the Mornington Peninsula,and the Grampians[Astroloma conostephioides(Sond.) Benth.,A.humifusum(Cav.) R. Br.,A pinifolium(R. Br.) Benth,Brachylomadaphnoides(Smith) Benth.,E. impressa, E. impressavar.grandifloraBenth.andStyphelia adscendensR. Br.] in western Victoria. For morphologicalstudies, samples of roots from each species at each site werecleared and stained and examined microscopically. For cross-inoculationstudies, cuttings from each site were struck in potting mediuminoculated with soil from the same and other sites. The ericoidmycorrhizae in the roots of plants found at or grown in Cranbourneand Rye soils were similar. Both were significantly differentfrom the internal hyphae found in the roots of plants foundat or grown in Grampians soils, which were three times largerin diameter and formed dense coils which filled the host celland invaded adjacent epidermal cells. This suggests that morethan one fungus is involved in the relationships, that the MorningtonPeninsula sites had a different fungus from the Grampians siteand that host specificity is low. Vesicular structures werealso found commonly on plants at the Grampians site, in contrastwith other sites. Epacridaceae; root; fungus; mycorrhiza; morphology; inoculation     

Characterization of amylolytic and pullulytic enzymes from thermophilic archaea and from a newFervidobacterium species

Nine extremely thermophilic archaea and one novel thermophilic bacterium were screened for their ability to produce amylolytic and pullulytic enzymes. Cultivation of these micro-organisms was performed in the absence of elemental sulphur with starch as the major carbon source. Enzymatic activity was mainly detected in two archaea belonging to the order Thermoproteales,Desulfurococcus mucosus andStaphylothermus marinus, in two archaea belonging to the order Thermococcales,Thermococcus celer andT. litoralis and in two novel archaeal strains, TYS and TY previously isolated from the Guaymas Basin in the Gulf of California. Both amylolytic and pullulytic activities were also detected in a newly isolated thermophilic bacterium belonging to the order Thermotogales and previously described asFervidobacterium pennavorans. Best yields for enzyme production were obtained in 1–1 batch cultures with the strains TYS (13 units U/1 of amylase, 6 U/1 of pullulanase),F. pennavorans (2.5 U/l of amylase, 4.5 U/l of pullulanase) andT. litoralis (3.0 U/l of amylase). Enzymes were in general characterized by temperature optima around 90–100°C, pH optima around 5.5–6.5 and a high degree of thermostability. Due to the remarkable properties of these enzymes, they are of interest for biotechnological applications.     

Water potentials for developing cladodes and fruits of a succulent plant, including xylem-versus-phloem implications for water movement

Developing cladodes had lower water potentials and developingfruits had higher water potentials than the underlying cladodesof the widely cultivated prickly pear cactus, Opuntia ficus-indica.The 0.06 MPa lower value in 4-week-old daughter cladodes indicateda typical water potential gradient from the underlying clad-odealong the xylem of –0.2 MPa m^–1; the 0.17 MPa highervalue in 4-week-old fruits, which decreased to 0.07 MPa by 10weeks, implicated the phloem as their supplier of water. Thephloem sap of the underlying cladodes had an osmotic pressureof only 0.90 to 0.98 MPa, so the phloem could supply a relativelydilute solution to the photosynthetically dependent fruits (daughtercladodes of O. ficus-indica are photosynthetically independentat 4 weeks). Although the water potentials were similar foradjacent tissues, the osmotic pressures were lower for the water-storagecompared with the photosynthetic tissue; the osmotic pressureswere higher for xylem sap from fruits, for which xylary flowapparently occurred toward the underlying cladodes, than fordaughter cladodes. The relative capacitance (change in relativewater content divided by change in tissue water potential) wasapproximately 0.71 MPa^–1 for the water-storage tissueand the photosynthetic tissue of both daughter cladodes andfruits at 4 weeks of age. When these organs approached maturityat 10 weeks, the relative capacitance increased about 40% fortheir water-storage tissue, but decreased 30% for their photosynthetictissue. As the plant water content decreases during drought,about twice as much water will thus be lost per unit volumeof the water-storage tissue compared with the photosynthetictissue of maturing fruits and cladodes. Key words: Opuntia ficus-indica, phloem, relative water content, water capacitance, water potential     

Human T Cell Leukemia Virus Type 1 (HTLV-1) Antibodies in Healthy Populations and Renal Transplanted Patients in the North-East of Brazil

The seroprevalence of human T cell leukemia virus type 1 (HTLV-1) infection was investigated in Brazilians (570): native inhabitants (298) and descendants from Japanese (272) living in Recife and its neighborhoods—North-east of Brazil. Furthermore, polytransfused renal transplanted patients (54) were also examined for the serological status to this virus. The seropositivity to HTLV-1, screened by enzyme-linked immunosorbent assay (ELISA), was low: 1.34% for the local population and 0.73% for the descendants from Japanese. However, the seropositivity for the renal transplanted patients was found to be 11.1%. This higher value suggests that this retrovirus infection seems to be of importance in this clinical condition.     

Effects of Temperature on Stereochemistry of Enzymatic Reactions

A brief discussion of the theoretical basis for effects of temperature on stereoselectivity of enzyme catalysed reactions is presented. In theory, the stereoselectivity of an enzymatic reaction can either increase or decrease as the reaction temperature is raised. The secondary alcohol dehydrogenase from Thermoanaerobacter ethanolicus reduces 2-butanone to (R)-2-butanol at 37° C, with increased stereoselectivity at higher temperatures and in the presence of NADP analogues. In contrast, at 37°, 2-pentanone and 2-hexanone are reduced to (S)-2-pentanol and (S)-2-hexanol, respectively, but the stereoselectivity decreases at higher temperatures and in the presence of NADP analogues. Reduction of racemic 2-methylbutanal by the primary alcohol dehydrogenase from T. ethanolicus gives (S)-2-methyl-1-butanol with greater stereospecificity at 35° (51% e.e.) than at 15° (14% e.e.). Horse liver alcohol dehydrogenase shows a preference for oxidation of the (S)-enantiomers of acyclic secondary alcohols at 25°, with a decrease in stereospecificity at higher temperatures.     

Modeling Lanthanide Complexes: Towards the Theoretical Design of Light Conversion Molecular Devices

Theoretical techniques have been developed and/or improved to predict the molecular structure of lanthanide complexes which were used to calculate their electronic properties, in particular, their electronic spectra and energy levels necessary to calculate the rates of energy transfer from the ligands to the metal ion. The molecular structure has been obtained by the SMLC/AM1 (Sparkle Model for the Calculation of Lanthanide Complexes – Austin Model 1) model where the lanthanide ion is simulated by a sparkle implemented into the AM1 Hamiltonian used to perform a HF-SCF (Hartree-Fock Self-Consistent Field) calculation. The previous implementation of the SMLC/AM1 model (sparkle/1) involving only two parameters has been generalized to be consistent with the AM1 Hamiltonian and the new model (sparkle/2) significantly improved the prediction of molecular structures of Eu(III) complexes. For the electronic spectra and energy level calculations of the lanthanide complexes the model replaces the metal ion by a point charge with the ligands held in their positions as determined by the SMLC/AM1 model, and uses a INDO/S-CI (intermediate neglect of differential overlap/spectroscopic-configuration interaction) model. A preliminary study of the solvent effects on the absorption spectra of the free ligand is also presented. For the ligand-lanthanide ion energy transfer Fermi's golden rule is used with the multipolar and exchange mechanisms being implemented and tested for several complexes. These theoretical techniques have been applied to several complexes yielding very good results when compared to experimental data as well as predictions for the molecular and electronic structures and the relative contributions of the mechanisms for the energy transfer rates. This revised version was published online in June 2006 with corrections to the Cover Date.     

Relationship of the apolipoprotein E polymorphism with carotid artery atherosclerosis.

From the cohort taking part in the Atherosclerosis Risk in Communities (ARIC) study, a multicenter investigation of atherosclerosis and its sequelae in women and men ages 45-64 years, a sample of 145 subjects with significant carotid artery atherosclerosis but without clinically recognized coronary heart disease was identified along with 224 group-matched control subjects. The aim of this paper is to measure the association of the apolipoprotein (apo) E polymorphism with the prevalence of significant carotid artery atherosclerotic disease (CAAD) after considering the contribution of established risk factor variables. The first model used a stepwise selection procedure to define a group of significant physical and lifestyle characteristics and a group of significant plasma lipid, lipoprotein, and apolipoprotein variables that were predictive of CAAD status in this sample. Those variables selected included age (years), body mass index (BMI; kg/m2), consumption of cigarettes (CigYears; number of cigarettes/d x the number of smoking years), hypertension status, high-density lipoprotein (HDL)-cholesterol (mg/dl), total cholesterol (mg/dl), and Lp[a] (micrograms/ml). The second model was built by forcing into the equation an a priori set of demographic, anthropometric, and lipoprotein variables, which were age, BMI, CigYears, hypertensive status, LDL-cholesterol, and HDL-cholesterol. In both models, the apo E genotype epsilon 2/3 was related to CAAD status. For both models, the estimated odds ratio of being a CAAD case associated with the apo E genotype epsilon 2/3 was > 2:1. The mechanism of the observed association between the epsilon 2/3 genotype and carotid atherosclerosis is unknown, but it is likely due to the known effects of the E2 isoform in causing delayed clearance of triglyceride-rich lipoproteins.     

Changes in cell-surface carbohydrates of Trypanosoma cruzi during metacyclogenesis under chemically defined conditions.

Highly purified lectins with specificities for receptor molecules containing sialic acid, N-acetylglucosamine (D-GlcNAc), N-acetylgalactosamine (D-GalNAc), galactose (D-Gal), mannose-like residues (D-Man) or L-fucose (L-Fuc), were used to determine changes in cell-surface carbohydrates of the protozoal parasite Trypanosoma cruzi during metacyclogenesis under chemically defined conditions. Of the D-GalNAc-binding lectins, BS-I selectively agglutinated metacyclic trypomastigotes, MPL was selective for replicating epimastigotes, whereas SBA strongly agglutinated all developmental stages of T. cruzi. WGA (sialic acid and/or D-GlcNAc specific) was also reactive with differentiating epimastigotes and metacyclic trypomastigotes but displayed a higher reactivity with replicating epimastigote forms. A progressive decrease in agglutinating activity was observed for jacaline (specific for D-Gal) during the metacyclogenesis process; conversely, a progressive increase in affinity was observed for RCA-I (D-Gal-specific), although the reactivity of other D-Gal-specific lectins (PNA and AxP) was strong at all developmental stages. All developmental stages of T. cruzi were agglutinated by Con A and Lens culinaris lectins (specific for D-Man-like residues); however, they were unreactive with the L-fucose-binding lectins from Lotus tetragonolobos and Ulex europaeus. These agglutination assays were further confirmed by binding studies using 125I-labelled lectins. Neuraminidase activity was detected in supernatants of cell-free differentiation medium using the PNA hemagglutination test with human A erythrocytes. The most pronounced differences in lectin agglutination activity were observed between replicating and differentiating epimastigotes, suggesting that changes in the composition of accessible cell-surface carbohydrates precede the morphological transformation of epimastigotes into metacyclic trypomastigotes.