Tau-related protein present in paired helical filaments has a decreased tubulin binding capacity as compared with microtubule-associated protein tau

We have isolated, after exhaustive detergent treatments, a 33 kDa tau-related protein isolated from paired helical filaments from Alzheimer's disease patient brains. The N-terminal sequence of the 33 kDa protein begins at residue 71 of the sequence described for human fetal tau protein. This truncated form of tau is not the consequence of the translation of a tau RNA lacking a region at its 5' end, as measured by primer extension analyses, suggesting that the 33 kDa protein must be generated by proteolysis of previously synthesized tau. This tau-related protein has only one blocked cysteine residue and also has a decreased tubulin binding capacity as compared with that of tau protein.     

Comparison of the hydrological characteristics of three small experimental holm oak forested catchments in NE Spain in relation to larger areas

Precipitation and streamflow have been measured in three small (0.04–0.52 km²) experimental catchments covered by dense holm oak (Quercus ilex L.) forests. Two of them are in the Prades mountains and one in the Montseny mountains (NE Spain). Here we test the hydrological representativeness of these catchments against the streamflow record at two nearby larger (34–60 km²) catchments, one from each massif. Comparisons of (i) annual streamflow, (ii) seasonal distribution of streamflow, and (iii) flow duration curves were made. At Prades, for the period of common record, mean annual precipitation was about 580 mm, and mean annual streamflow 44–81 mm at the two experimental catchments and 102 mm at the larger one. Most streamflow occurred during winter and spring in the three catchments. At Montseny, rainfall was higher, and mean annual streamflow was 495 mm in the experimental catchment, and 760 mm in the larger catchment, though these data were obtained in different periods in each catchment. Streamflow was roughly equal in autumn, winter and spring. At both sites flow duration curves were fairly similar in the small experimental catchments and the larger catchments. The higher streamflow at Montseny is reflected in its flow duration curves being well above those at Prades. The experimental catchments at Prades are thus fairly representative of the larger nearby catchment for the investigated hydrological characteristics. At Montseny, hydrological differences between the experimental catchment and the larger catchment are probably due to the higher mean altitude of the latter and to the non-overlapping periods of their streamflow records.     

Marinalg International Awards

Marinalg International Awards     

General organization of the conjugal transfer genes of the IncW plasmid R388 and interactions between R388 and IncN and IncP plasmids.

The complete conjugal transfer gene region of the IncW plasmid R388 has been cloned in multicopy vector plasmids and mapped to a contiguous 14.9-kilobase segment by insertion mutagenesis. The fertility of the cloned region could still be inhibited by a coresident IncP plasmid. The transfer region has been dissected into two regions, one involved in pilus synthesis and assembly (PILW), and the other involved in conjugal DNA metabolism (MOBW). They have been separately cloned. PILW also contains the genes involved in entry exclusion. MOBW contains oriT and the gene products required for efficient mobilization by PILW. MOBW plasmids could also be mobilized efficiently by PILN, the specific pilus of the IncN plasmid pCU1, but not by PILP, the specific pilus of the IncP plasmid RP1.     

Trypanosoma cruzi: allopurinol in the treatment of mice with experimental acute Chagas disease

The therapeutic effect of allopurinol was studied in an experimental Trypanosoma cruzi infection (Chagas disease) in outbred IVIC-NMRI and inbred C57B1/6J mice intraperitoneally inoculated with the parasites 2–6 days before drug treatment. Allopurinol protected against T. cruzi infection. This effect was evidenced by highly significant reductions in both parasitemias and mortality rates and increased survival time in allopurinol-treated animals compared with untreated infected mice. Allopurinol protected effectively when administered in 10 daily doses of 32–64 mg/kg body wt/day injected intraperitoneally. Using direct methods, parasitemia remained undetectable for at least 310 days. An indirect method, subinoculation to susceptible mice, showed a few circulating trypanosomes which decreased greatly in number after a second schedule of allopurinol treatment; finally no trypanosomes were detectable 275 days after treatment initiation. Allopurinol also induced a strong trypanostatic effect when tested in vitro on five different Trypanosoma cruzi strains (optimal inhibitory concentration: 3 μg/ml). These results suggest that allopurinol protects mice with acute Chagas infection by a direct trypanostatic effect. The low toxicity of this drug suggests its use in more chronic experimental Chagas infections.     

Trypanosoma cruzi: defined medium for continuous cultivation of virulent parasites.

A liquid medium was developed for the continuous cultivation of Trypanosoma cruzi. Among the several highly purified macromolecules tested only bovine liver catalase, horseradish peroxidase, lactoperoxidase, and bovine hemoglobin supported the continuous growth, at high yield, of mice-virulent Trypanosoma cruzi; other hemoproteins were inactive. Bovine liver catalase showed optimal Trypanosoma cruzi growth-promoting activity, parasites reaching 20 × 10⁶ parasites/ml (95% epimastigotes) at about 10 days in most of the 45 subpassages to date. Furthermore, this protein in the incubation medium provided all the amino acid requirements of actively growing parasites, thus eliminating the need for exogeneous free amino acids. Additional experiments revealed that the hemoprotein's growth-promoting activity was independent of any enzymatic activity and that reconstituting the exact protein composition by means of exogeneous amino acids did not support parasite multiplication, suggesting the importance of the primary structure of the active proteins for growth-promoting activity. These active macromolecules supported the multiplication of five different strains of Trypanosoma cruzi, but did not support Leishmania brasiliensis or Leishmania mexicana proliferation, suggesting species specificity.     

Binding of microtubule proteins to DNA: specificity of the interaction.

Tubulin is detected among the DNA-binding proteins when an extract from fibroblasts is chromatographed on DNA-cellulose. Further purification of the colchicine-binding activity shows that purified tubulin from fibroblasts does not bind to DNA. Depolymerized brain microtubule proteins show a high affinity for DNA. The fraction bound is composed of tubulin and microtubule-associated proteins. Experiments with fractionated microtubule proteins indicate that tubulin-free microtubule associated proteins bind to DNA, while tubulin free of microtubule-associated proteins does not. Microtubule-associated proteins bind better to eukaryotic than to phage DNA suggesting a specificity of the interaction.     

Alkylguanidines as inhibitors of k transport in isolated barley roots

It has been shown that plants can accumulate K⁺ through an energy-dependent process. The effect of alkylguanidines, in particular octylguanidine on the uptake of ⁸⁶Rb⁺ by excised barley roots (Hordeum vulgare var. Apizaco LV-72), has been studied. ⁸⁶Rb⁺ was used as tracer of K⁺. The uptake of ⁸⁶Rb⁺ which is linear with time and shows saturation kinetics is inhibited by octylguanidine. Half-maximal inhibition of ⁸⁶Rb⁺ uptake is attained at 50 μM octylguanidine. Octylguanidine induces a decrease in the V_max of the process and increases the Km of the system for Rb⁺. When the effects of various alkylguanidines were studied, the following order of effectiveness was encountered; octylguanidine = hexilguanidine > butylguanidine > ethylguanidine > guanidine. This suggests that guanidines inhibit Rb⁺ uptake by interacting through its positively charged guanidinium group with a Rb⁺ carrier while the alkyl chain interacts with the hydrophobic milieu of the membrane.     

A new tritoniid species (Mollusca: Opisthobranchia) from Bouvet Island

Tritoniid sea slugs (Mollusca, Gastropoda, Opisthobranchia) are reported for the first time from Bouvet Island. Tritonia dantarti sp. n. shows morphological and anatomical differences with regard to the two previously known tritoniids reported from Antarctica and Sub-Antarctica, Tritonia vorax (Odhner 1926) and Tritonia challengeriana Bergh, 1884. Regarding the external morphology, T. dantarti sp. n. is characterized by a very bright orange coloration in the dorsum, white dorsal crests, highly ramified dendritic gills (the largest vertically orientated and the rest laterally orientated), and a quadrangular cross section. The radula presents very long, thin lateral teeth and the jaws present mainly unicuspidate, striated denticles. The seminal receptacle is large, pear-shaped and its grey pigmentation differs from the rest of the genital system. The Antarctic species T. challengeriana was also found in waters off Bouvet Island, while the Subantarctic species T. vorax was not found.     

Effects of flask sealing and growth regulators on in vitro propagation of neem (Azadirachta indica A. Juss.)

In vitro propagation of neem (Azadirachta indica A. Juss.) may offer an efficient alternative to seed propagation of this species. For optimization of in vitro propagation, different basal salt formulations, growth regulators, and culture container sealants (polytetrafluoroethylene hydrophobic membranes [PTFE]) were evaluated. Nodal segments cultured on Murashige and Skoog (MS) medium showed the highest shoot formation per explant (1.67). Explants cultured in flasks containing MS medium with 0.5 mg L⁻¹ benzyladenine, 0.5 mg L⁻¹ kinetin, and 0.05 mg L⁻¹ naphthaleneacetic acid, and sealed with two PTFE membranes, produced the highest number of shoots (4.04). In contrast, explants cultured in flasks without membranes showed leaf chlorosis and senescence. For plant recovery, regenerants were acclimatized in a substrate of coconut fiber and eucalyptus bark (1:1) and showed 80% survival. Our results indicated that the use of flasks with vents was beneficial for in vitro propagation of this important plant.