Discrimination Amongst Leishmania by Polymerase Chain Reaction and Hybridization with Small Subunit Ribosomal DNA Derived Oligonucleotides

ABSTRACT. A method for discriminating among Leishmania is described, based upon small subunit ribosomal DNA sequence differences. The method was to amplify the entire 2.2 kb small subunit rDNA by polymerase chain reaction using conserved primers specific for the 5' and 3' termini of the small subunit ribosomal RNA, and then hybridize the product dotted onto nylon membranes with labeled oligonucleotides. The design of the hybridization probes was based upon complete small subunit rDNA sequences from L. amazonensis, L. major and L. guyanensis and partial sequences of L. mexicana, L. braziliensis, L. tropica and L. chagasi. A high degree of sequence similarity (> 99%) among species was found. However, sufficient sequence divergence occurred to permit the design of internal oligonucleotide probes specific for species complexes. This procedure successfully discriminated amongst a wide range of Leishmania isolates. The method detected as few as 10 cultured organisms and detected parasites in tissue samples from experimentally infected animals. Non-radioactive labeling showed the same specificity and sensitivity as radioactive probes.     

Reproductive characters in a gynodioecious species, Silene italica (Caryophyllaceae), with attention to the gynomonoecious phenotype

Gynodioecious populations (i.e. populations with female and hermaphrodite individuals) often contain a third phenotype with an intermediate sex expression. In Silene italica , this phenotype is characterized by a mixture of pistillate and perfect flowers and is thus gynomonoecious. To characterize sexual functions of these gynomonoecious individuals and their potential influence in the maintenance of gynodioecy, reproductive characters of the three sexual phenotypes were compared over 2 years in several families of S. italica produced by crossing. We found that gynomonoecious individuals were intermediate for flower production and female fertility characters, although they did not always significantly differ from female individuals. Perfect flowers of gynomonoecious and hermaphrodite plants were similar in size and pollen production. Gynomonoecious individuals were thus intermediate in female and in male functions. Family effects were found for most of the characters. The female advantage (i.e. the fertility of females compared to the female fertility of pollen producing plants) was not dramatically different when gynomonoecious plants were taken into account.  © 2006 The Linnean Society of London, Biological Journal of the Linnean Society , 2006, 87 , 583–591.     

Active Transport of Glutamate in Leishmania (Leishmania) amazonensis

ABSTRACT. Leishmania spp. are the causative agents of leishmaniasis, a complex of diseases with a broad spectrum of clinical manifestations. Leishmania (Leishmania) amazonensis is a main etiological agent of diffuse cutaneous leishmaniasis. Leishmania spp., as other trypanosomatids, possess a metabolism based significantly on the consumption of amino acids. However, the transport of amino acids in these organisms remains poorly understood with few exceptions. Glutamate transport is an important biological process in many organisms. In the present work, the transport of glutamate is characterized. This process is performed by a single kinetic system (K_m=0.59±0.04 mM, V_max=0.123±0.003 nmol/min per 20 × 10⁶ cells) showing an energy of activation of 52.38±4.7 kJ/mol and was shown to be partially inhibited by analogues, such as glutamine, aspartate, α‐ketoglutarate and oxaloacetate, methionine, and alanine. The transport activity was sensitive to the extracellular concentration of H⁺ but not to Na⁺ or K⁺. However, unlike other amino acid transporters presently characterized, the treatment with specific ionophores confirmed the participation of a K⁺, and not H⁺ membrane gradient in the transport process.     

The interaction of soil biota and soil structure under global change

The structural framework of soil mediates all soil processes, at all relevant scales. The spatio-temporal heterogeneity prevalent in most soils underpins the majority of biological diversity in soil, providing refuge sites for prey against predator, flow paths for biota to move, or be moved, and localized pools of substrate for biota to multiply. Just as importantly, soil biota play a crucial role in mediating soil structure: bacteria and fungi aggregate and stabilize structure at small scales (μm–cm) and earthworms and termites stabilize and create larger-scale structures (mm–m). The stability of this two-way interaction of structure and biota relations is crucial to the sustainability of the ecosystem. Soil is constantly reacting to changes in microclimates, and many of the soil–plant–microbe processes rely on the functioning of subtle chemical and physical gradients. The effect of global change on soil structure–biota interactions may be significant, through alterations in precipitation, temperature events, or land-use. Nonetheless, because of the complexity and the ubiquitous heterogeneity of these interactions, it is difficult to extrapolate from general qualitative predictions of the effects of perturbations to specific reactions. This paper reviews some of the main soil structure–biota interactions, particularly focusing on soil stability, and the role of biota mediating soil structures. The effect of alterations in climate and land-use on these interactions is investigated. Several case studies of the effect of land-use change are presented.