Reduction of the metallochromic indicators arsenazo III and antipyrylazo III to their free radical metabolites by cytoplasmic enzymes

At a concentration much lower than that usually employed for measuring cytosolic ionized Ca²⁺ concentrations, arsenazo III underwent a one-electron reduction by rat liver cytosolic fraction or a hypoxanthinexanthine oxidase system to produce an azo anion radical metabolite. NADH, NADPH, N¹-methylnicotinamide, hypoxanthine, and xanthine, in that order, could serve as a source of reducing equivalents for the production of this free radical by the cytosolic fraction. The steady-state concentration of the azo anion radical and the arsenazo III-stimulated O₂ consumption were enhanced by calcium and magnesium. Antipyrylazo III was ineffective in increasing O₂ consumption by rat liver cytosolic fraction and gave a much weaker ESR signal of an azo anion radical with both the liver cytosolic fraction, in the presence of NADH, and the hypoxanthine-xanthine oxidase system.     

Increased replication in plant nuclei, whose spread chromatin contains enlarged nucleosomal linkers, is only partially due to a rise in the number of DNA elongating chains

Dormant and proliferating meristems of Allium cepa L. roots were compared. In vivo DNA synthesis was nine times higher in proliferating meristems, partially due to a 2.5 times increase in the relative number of DNA elongating chains, as estimated after the assay for 3H-TTP incorporation in permeabilized cells. Stereology done on conventional electronmicrographs showed that nucleoplasm was nearly three times larger in nuclei of proliferating meristems, while the amount of compact chromatin did not diminish but redistributed instead, resulting that the chromatin/nucleoplasm interface increased. Finally, the nucleosomal linker -as seen after the spreading of the chromatin fibres- was larger and more variable in proliferating than in dormant meristems.     

Validation of data collection for an American trypanosomiasis epidemiologic study in the State of Morelos, México]

The purpose of this paper is to show up the importance of the standardization concepts in American Trypanosomiasis epidemiological studies. The consistence in the measurement of some dwelling characteristics was evaluated. A validation of the Queretaro antigen for indirect hemagglutination reaction as a diagnostic test and the interobserver concordance for the serologic readings were also made. The observers were instructed in some sessions. The pretests were made in the laboratory with positive and negative sera, with sera from the studied population. Results show that the interobserver concordance after the instruction, for the dwelling variables ranged from 70% to 100%. Sensitivity of the Queretaro antigen was 100%, specificity 55%, the predictive value of a positive test 55%, and the predictive value of a negative test 93%. The interobserver concordance was 47%. The pretest and the pilot study are very important in getting the objectives of the principal study.     

Brucella abortus 16S rRNA and lipid A reveal a phylogenetic relationship with members of the alpha-2 subdivision of the class Proteobacteria

On the basis of ribosomal 16S sequence comparison, Brucella abortus has been found to be a member of the alpha-2 subdivision of the class Proteobacteria (formerly named purple photosynthetic bacteria and their nonphototrophic relatives). Within the alpha-2 subgroup, brucellae are specifically related to rickettsiae, agrobacteria, and rhizobiae, organisms that also have the faculty or the obligation of living in close association to eucaryotic cells. The composition of Brucella lipid A suggests a close phylogenetical relationship with members of the alpha-2 group. The chemical analysis of the lipid A fraction revealed that Brucella species contain both glucosamine and diaminoglucose, thus suggesting the presence of a so-called mixed lipid A type. The serological analysis with polyclonal and monoclonal antibodies is in agreement with the existence of mixed lipid A type in B. abortus. The amide-linked fatty acid present as acyl-oxyacyl residues were 3-O-C(16:0)12:0, 3-O-C(16:0)13:0, 3-O-C(16:0)14:0, and 3-O-C(18:0)14:0. The only amide-linked unsubstituted fatty acid detected was 3-OH-C16:0. The ester-linked fatty acids are 3-OH-C16:0, 3-OH-C18:0, C16:0, C17:0, and C18:0. Significant amounts of the large-chain 27-OH-C28:0 were detected together with traces of 25-OH-C26:0 and 29-OH-C30:0. Comparison of the Brucella lipid composition with that of the other Proteobacteria also suggests a close phylogenetical relationship with members of the alpha-2 subdivision. The genealogical grouping of Brucella species with pericellular and intracellular plant and animal pathogens as well as with intracellular plant symbionts suggests a possible evolution of Brucella species from plant-arthropod-associated bacteria.     

Oligomerization of the reversibly glycosylated polypeptide: its role during rice plant development and in the regulation of self-glycosylation

A multigenic family of self-glycosylating proteins named reversibly glycosylated polypeptides, designated as RGPs, have been usually associated with carbohydrate metabolism, although they are an enigma both at the functional, as well as at the structural level. In this work, we used biochemical approaches to demonstrate that complex formation is linked to rice plant development, in which class 1 Oryza sativa RGP (OsRGP) would be involved in an early stage of growing plants, while class 2 OsRGP would be associated with a late stage linked to an active polysaccharide synthesis that occurs during the elongation of plant. Here, a further investigation of the complex formation of the Solanum tuberosum RGP (StRGP) was performed. Results showed that disulfide bonds are at least partially responsible for maintaining the oligomeric protein structure, so that the nonreduced StRGP protein showed an apparent higher molecular weight and a lower radioglycosylation of the monomer with respect to its reduced form. Hydrophobic cluster analysis and secondary structure prediction revealed that class 2 RGPs no longer maintained the Rossman fold described for class 1 RGP. A 3D structure of the StRGP protein resolved by homology modeling supports the possibility of intercatenary disulfide bridges formed by exposed cysteines residues C79, C303 and C251 and they are most probably involved in complex formation occurring into the cell cytoplasm.     

Primary and secondary vegetation patches as contributors to floristic diversity in a tropical deciduous forest landscape

The floristic diversity of Mexican tropical deciduous forests (TDF) is of critical importance given the high species richness (alpha diversity), species turnover (beta diversity), and the intense deforestation rates. Currently, most TDF landscapes are mosaics of agricultural land, secondary vegetation, and patches of relatively undisturbed primary vegetation. Here we illustrate how both primary forest remnants and secondary vegetation patches contribute to the floristic diversity of TDF in a landscape of volcanic origin in central Veracruz, Mexico. Our objectives were to assess sampling efficiency and inventory completeness, to compare mean and cumulative species richness between primary forest and secondary vegetation sites, and to analyze beta diversity between vegetation types. In an area of 12,300 m2 we recorded 105 families, 390 genera, and 682 species. Species inventories for both vegetation types were about 80% complete. Secondary vegetation is more alpha diverse than primary forest, both in terms of cumulative and mean species richness. We found a remarkably high beta diversity between vegetation types (75% of complementarity, 91.60% of mean dissimilarity). We also identified the species that contribute the most to similarity within vegetation types and to dissimilarity between vegetation types. Our results support the idea that assessing biodiversity on the landscape scale is an appropriate way to ascertain the impact of human activities. For this land mosaic, conservation of the flora would not be possible by focusing solely on primary forest remnants. We propose the implementation of a network of small conservation areas with a flexible structure, following the “archipelago reserve” model.