Interaction between ion channel-inactivating peptides and anionic phospholipid vesicles as model targets.

Studies of rapid (N-type) inactivation induced by different synthetic inactivating peptides in several voltage-dependent cation channels have concluded that the channel inactivation "entrance" (or "receptor" site for the inactivating peptide) consists of a hydrophobic vestibule within the internal mouth of the channel, separated from the cytoplasm by a region with a negative surface potential. These protein domains are conformed from alternative sequences in the different channels and thus are relatively unrestricted in terms of primary structure. We are reporting here on the interaction between the inactivating peptide of the Shaker B K+ channel (ShB peptide) or the noninactivating ShB-L7E mutant with anionic phospholipid vesicles, a model target that, as the channel's inactivation "entrance," contains a hydrophobic domain (the vesicle bilayer) separated from the aqueous media by a negatively charged vesicle surface. When challenged by the anionic phospholipid vesicles, the inactivating ShB peptide 1) binds to the vesicle surface with a relatively high affinity, 2) readily adopts a strongly hydrogen-bonded beta-structure, likely an intramolecular beta "hairpin," and 3) becomes inserted into the hydrophobic bilayer by its folded N-terminal portion, leaving its positively charged C-terminal end exposed to the extravesicular aqueous medium. Similar experiments carried out with the noninactivating, L7E-ShB mutant peptide show that this peptide 1) binds also to the anionic vesicles, although with a lower affinity than does the ShB peptide, 2) adopts only occasionally the characteristic beta-structure, and 3) has completely lost the ability to traverse the anionic interphase at the vesicle surface and to insert into the hydrophobic vesicle bilayer. Because the negatively charged surface and the hydrophobic domains in the model target may partly imitate those conformed at the inactivation "entrance" of the channel proteins, we propose that channel inactivation likely includes molecular events similar to those observed in the interaction of the ShB peptide with the phospholipid vesicles, i.e., binding of the peptide to the region of negative surface potential, folding of the bound peptide as a beta-structure, and its insertion into the channel's hydrophobic vestibule. Likewise, we relate the lack of channel inactivation seen with the mutant ShB-L7E peptide to the lack of ability shown by this peptide to cross through the anionic interphase and insert into the hydrophobic domains of the model vesicle target.     

Past vegetation changes in the Brazilian Pantanal arboreal–grassy savanna ecotone by using carbon isotopes in the soil organic matter

Measurements of the organic carbon inventory, its stable isotopic composition and radiocarbon content were used to deduce vegetation history from two soil profiles in arboreal and grassy savanna ecotones in the Brazilian Pantanal. The Pantanal is a large floodplain area with grass-dominated lowlands subject to seasonal flooding, and arboreal savanna uplands which are only rarely flooded. Organic carbon inventories were lower in the grassy savanna site than in the upland arboreal savanna site, with carbon decreasing exponentially with depth from the surface in both profiles. Changes in ¹³C of soil organic matter (SOM) with depth differed markedly between the two sites. Differences in surface SOM ¹³C values reflect the change from C₃ to C₄ plants between the sites, as confirmed by measurements of ¹³C of vegetation and the soil surface along a transect between the upland closed-canopy forest and lowland grassy savanna. Changes of ¹³C in SOM with depth at both sites are larger than the 3–4 per mil increases expected from fractionation associated with organic matter decomposition. We interpret these as recording past changes in the relative abundance of C₃ and C₄ plants at these sites. Mass balances with ¹⁴C and ¹³C suggest that past vegetational changes from C₃ to C₄ plants in the grassy savanna, and in the deeper part of the arboreal savanna, occurred between 4600 and 11 400 BP, when major climatic changes were also observed in several places of the South American Continent. The change from C₄ to C₃, observed only in the upper part of the arboreal savanna, was much more recent (1400 BP), and was probably caused by a local change in the flooding regime.     

HYDROBIIDAE (GASTROPODA: HYDROBIOIDEA) FROM THE RIBEIRA VALLEY, S.E. BRAZIL, WITH DESCRIPTIONS OF TWO NEW CAVERNICOLOUS SPECIES

Potamolithus karsticus n. sp. and Potamolithus troglo-bius n.sp., two Brazilian aquatic cavesnails (Gastropoda: Hydrobiidae),are described. P. troglobius is blind and depigmented, and isthe first stygobiontic snail to be described from Brazil Additionally, specimens of Potamolithus ribeirensis Pilsbry,1911 were collected near the type locality for comparison withthe new cave species (Received 2 December 1993; accepted 20 June 1994)     

Phylogenetic relationships of the garter snakes based on DNA sequence and allozyme variation

We estimated phylogenetic relationships among 26 species of garter snakes (genus Thamnophis ) using allozyme and mitochondrial cytochrome b gene nucleotide sequence variation. Parsimony analyses of the two data sets give substantially different estimates of phylogeny. Several lines of evidence indicate that much of this conflict is due to error associated with the restricted number of characters in each data set. Such sampling error may be reduced by combining all the characters; we therefore present an estimate of phylogeny based on parsimony analysis of all the data combined. All our analyses support several conclusions in conflict with previous views: a very distant relationship between T.errans and T. elegans , non-monophyly of the elegans group (even excluding T: errans ), and nesting of the form validus (previously considered a member of the genus Nerodia ) within Thamnophis.
The combined analysis gives an almost fully resolved tree. However, bootstrapping indicates only weak support for many clades in this tree. Furthermore, paraphyly of the assemblages of cytochrome b gene lineages within T. elegans and T. radix indicate the potential for discordance between the mitochondrial DNA (mtDNA) and species phylogenies through the sorting of ancestral mtDNA polymorphisms. These problems suggest the need for assaying additional characters, especially ones likely to be independent of those used in the present study.     

Effect of Sodium on Phosphate Uptake in Unicellular and Filamentous Cyanobacteria

The effect of Na⁺ on phosphate uptake was studied in four strainsof cyanobacteria: Synechococcus PCC 7942, Gloeothece PCC 6501,Phormidium sp. and Chlorogloeopsis PCC 6912. Phosphate uptakewas stimulated by Na⁺ in all cases. Li⁺ and K⁺ acted as partialanalogues for Na⁺. Half-saturation [K_1/2(Na⁺)] of phosphateuptake was reached with Na⁺ concentrations ranging from 317µM in Chlorogloeopsis to 659 µM in Phormidium. Theconcentration of phosphate required to reach half-saturationof phosphate uptake [K_1/2(P_i)]was not changed by the presenceof Na⁺. (Received April 11, 1994; Accepted July 5, 1994)     

Pleural Fluid Adenosine Deaminase and Lymphocyte Proportion: Clinical Usefulness In the Diagnosis of Tuberculosis

Adenosine deaminase (ADA) and lymphocyte proportion are known to be independently elevated in tuberculous effusions, but are non-specific, and false positive results are frequent. to overcome this problem the combined use of both parameters was prospectively studied in 276 patients with pleural effusion seen at Porto Alegre, Brazil. Using a cut-off level of 40 U/l at 37°C (method of Giusti¹⁹) for ADA activity and lymphocyte proportion of more than 50%, the correct diagnosis of tuberculosis (sensitivity) was made in 90.7% (CI 87.3–94.1%) of 54 patients. A specificity of 97.7% (CI 95.9–99.5%) was recorded. Five false positive diagnoses of tuberculous effusion were made. Five false negative diagnoses were made: three cases with haematogenous tuberculous dissemination with low ADA levels, and two other patients with low lymphocyte proportion. the combined use of ADA activity determination and lymphocyte proportion is a highly efficient diagnostic strategy of low cost, that merits wider use.