Identification of Trypanosoma cruzi Zymodemes by Kinetoplast DNA Probes

Analysis of zymograms of extracts of Trypanosoma cruzi isolated from different hosts in Argentina allowed characterization of 12 zymodemes or "isozymic strains," only six of which were found in human patients. Two of these six zymodemes (Z1 and Z12) were widely distributed and found in more than 80% of human patients. These two "major natural clones" differed significantly in pathogenic activity. Because the groupings obtained by studying enzymes and kinetoplast DNA (kDNA) were similar, it is possible to identify the zymodeme by analyzing kDNA. A 290-bp fragment was amplified by PCR using primers for the sequences flanking the hypervariable regions of kDNA minicircles. Labeled probes for this fragment, prepared from Z1 and Z12 reference stocks, hybridized specifically with PCR-amplified kDNA from parasite stocks, allowing identification of zymodemes.     

Projected changes in mineral soil carbon of European croplands and grasslands, 1990–2080

We present the most comprehensive pan‐European assessment of future changes in cropland and grassland soil organic carbon (SOC) stocks to date, using a dedicated process‐based SOC model and state‐of‐the‐art databases of soil, climate change, land‐use change and technology change. Soil carbon change was calculated using the Rothamsted carbon model on a European 10 × 10′ grid using climate data from four global climate models implementing four Intergovernmental Panel on Climate Change (IPCC) emissions scenarios (SRES). Changes in net primary production (NPP) were calculated by the Lund–Potsdam–Jena model. Land‐use change scenarios, interpreted from the narratives of the IPCC SRES story lines, were used to project changes in cropland and grassland areas. Projections for 1990–2080 are presented for mineral soil only. Climate effects (soil temperature and moisture) will tend to speed decomposition and cause soil carbon stocks to decrease, whereas increases in carbon input because of increasing NPP will slow the loss. Technological improvement may further increase carbon inputs to the soil. Changes in cropland and grassland areas will further affect the total soil carbon stock of European croplands and grasslands. While climate change will be a key driver of change in soil carbon over the 21st Century, changes in technology and land‐use change are estimated to have very significant effects. When incorporating all factors, cropland and grassland soils show a small increase in soil carbon on a per area basis under future climate (1–7 t C ha^?1 for cropland and 3–6 t C ha^?1 for grassland), but when the greatly decreasing area of cropland and grassland are accounted for, total European cropland stocks decline in all scenarios, and grassland stocks decline in all but one scenario. Different trends are seen in different regions. For Europe (the EU25 plus Norway and Switzerland), the cropland SOC stock decreases from 11 Pg in 1990 by 4–6 Pg (39–54%) by 2080, and the grassland SOC stock increases from 6 Pg in 1990 to 1.5 Pg (25%) under the B1 scenario, but decreases to 1–3 Pg (20–44%) under the other scenarios. Uncertainty associated with the land‐use and technology scenarios remains unquantified, but worst‐case quantified uncertainties are 22.5% for croplands and 16% for grasslands, equivalent to potential errors of 2.5 and 1 Pg SOC, respectively. This is equivalent to 42–63% of the predicted SOC stock change for croplands and 33–100% of the predicted SOC stock change for grasslands. Implications for accounting for SOC changes under the Kyoto Protocol are discussed.     

Nitrogen deposition interacts with climate in affecting production and decomposition rates in Sphagnum mosses

Increasing rates of atmospheric nitrogen (N) deposition may reduce growth and accelerate decomposition of Sphagnum mosses in bogs. Sphagnum growth and rates of Sphagnum litter decomposition may also vary because of climate change as both processes are controlled by climatic factors. The initial purpose of this study was to assess if growth and litter decomposition of hummock and lawn Sphagnum species varied with increasing N input in a factorial mid‐term (2002–2005) experiment of N and phosphorus (P) addition, in a bog on the southern Alps of Italy. However, as the experimental period was characterized by an exceptional heat wave in summer 2003, we also explored the interacting effects of fertilization and strongly varying climate on growth and decomposition rates of Sphagnum. The heat wave implied strong dehydration of the upper Sphagnum layer even if precipitation in summer 2003 did not differ appreciably from the overall mean. Sphagnum production was somewhat depressed by high levels (3 g m⁻² yr⁻¹) of N addition without concomitant addition of P presumably because of nutrient imbalance in the tissues, but production rates were much lower than the overall means in 2003, when no effect of nutrient addition could be observed. Adding N at high level also increased the potential decay of Sphagnum litter. Higher CO₂ emission from N‐fertilized litter was due to amelioration of litter chemistry showing lower C/N quotients in the N‐fertilized treatments. Rates of CO₂ emission from incubated litter also were more strongly affected by water content than by nutrient status, with practically no CO₂ emission detected when litter was dry. We conclude that higher rates of atmospheric N availability input may depress Sphagnum growth because of P, and presumably potassium, (co‐)limitation. Higher N availability is also expected to promote potential decay of Sphagnum litter by ameliorating litter chemistry. However, both effects are less pronounced if the growing Sphagnum apex and the underlying senescing tissues dry out.     

Over‐expression of bacterial γ‐glutamylcysteine synthetase (GSH1) in plastids affects photosynthesis,growth and sulphur metabolism in poplar (Populus tremula × Populus alba) dependent on the resulting γ‐glutamylcysteine and glutathione levels

We compared three transgenic poplar lines over‐expressing the bacterial γ‐glutamylcysteine synthetase (GSH1) targeted to plastids. Lines Lggs6 and Lggs12 have two copies, while line Lggs20 has three copies of the transgene. The three lines differ in their expression levels of the transgene and in the accumulation of γ‐glutamylcysteine (γ‐EC) and glutathione (GSH) in leaves, roots and phloem exudates. The lowest transgene expression level was observed in line Lggs6 which showed an increased growth, an enhanced rate of photosynthesis and a decreased excitation pressure (1‐qP). The latter typically represents a lower reduction state of the plastoquinone pool, and thereby facilitates electron flow along the electron transport chain. Line Lggs12 showed the highest transgene expression level, highest γ‐EC accumulation in leaves and highest GSH enrichment in phloem exudates and roots. This line also exhibited a reduced growth, and after a prolonged growth of 4.5 months, symptoms of leaf injury. Decreased maximum quantum yield (F_v/F_m) indicated down‐regulation of photosystem II reaction centre (PSII RC), which correlates with decreased PSII RC protein D1 (PsbA) and diminished light‐harvesting complex (Lhcb1). Potential effects of changes in chloroplastic and cytosolic GSH contents on photosynthesis, growth and the whole‐plant sulphur nutrition are discussed for each line.     

Interaction Between TRPC Channel Subunits in Endothelial Cells

Transient Receptor Potential Canonical (TRPC) proteins have been identified in mammals as a family of plasma membrane calcium-permeable channels activated by different kinds of stimuli in several cell types. We have studied TRPC subunit expression in bovine aortic endothelial (BAE-1) cells, where stimulation with basic fibroblast growth factor (bFGF), a potent angiogenetic factor, induces calcium entry carried at least partially by TRPC1 channels. By means of a RT-PCR approach, we have found that, in addition to TRPC1, only TRPC4 is expressed, both at the mRNA and protein level, as confirmed by immunoblotting and immunocytochemical analysis. Because functional TRPC channels are formed by assembly of four subunits in either homo- or heterotetrameric structures, we have carried out immunoprecipitation experiments and showed that TRPC1 and TRPC4 interact to form heteromers in these cells, independently from culture conditions (high or low percent of fetal calf serum, stimulation with bFGF). Moreover, the data show that TRPC subunits are not tyrosine-phosphorylated after bFGF stimulation and they do not co-immunoprecipitate with the type 1 FGF receptor. These results suggest that BAE-1 cells are a suitable model to study function and regulation of endogenous TRPC1/TRPC4 heteromers.     

The evolution of release and advertisement calls in green toads (Bufo viridis complex)

In the present paper we tested the hypothesis that differences in the acoustic communication system of diploid and tetraploid green toads ( Bufo viridis complex) might be due to selection for reproductive character displacement. We recorded two acoustic signals of the toad repertoire − the advertisement call (a long range mate-attracting signal) and the release call (a short-range signal mediating male–male interactions) − from six sympatric Central Asian populations (three diploid and three tetraploid populations) as well as from three allopatric diploid populations from Italy, and compared their patterns of variation with the pattern of among-population genetic distances. Although release and advertisement calls share the same morpho-physiological constraints, they show significantly different patterns of variation. Release calls vary congruently with the pattern of genetic distances, suggesting that mutation and genetic drift have been the major forces responsible for their change both in time and space. By contrast, the pattern of advertisement-call variation is not consistent with the phylogeny of the group, because the advertisement calls of Asian diploid and tetraploid populations differ from each other more than their genetic distances would predict. These results strongly support the hypothesis that selection acted on the advertisement calls of either or both Asian taxa , possibly, to favour reproductive isolation.  © 2002 The Linnean Society of London, Biological Journal of the Linnean Society , 2002, 77, 379–391.