Effect of Ethidium Bromide on the Oxidative Metabolism and Enzyme Profiles of Crithidia fasciculata*

SYNOPSIS. Changes in the metabolism of Crithidia fasciculata ATCC 11745 when grown in the presence of ethidium bromide were studied. Ethidium bromide-grown cells had decreased respiratory activity as measured by oxygen consumption. More than 50% of the organisms cultivated in a defined medium containing 1.0 mg/liter of ethidium bromide became dyskine-toplastic and had decreased activities of particulate succinate and NADH-linked dehydrogenases as well as of soluble isocitrate dehydrogenase. These cells also had increased activities of particulate α-glycerophosphate dehydrogenase, soluble α-glycerophosphate dehydrogenase, malic enzyme, hexokinase, and malate dehydrogenase. Ethidium bromide-grown cells had a lower level of ATP and contained less DNA than cells grown in its absence.     

Hierarchical genetic structure and gene flow in three sympatric species of Amazonian rodents

The population genetic structure of three species of Amazonian rodents ( Oligoryzomys microtis, Oryzomys capito , and Mesomys hispidus ) is examined for mtDNA sequence haplotypes of the cytochrome b gene by hierarchical analysis of variance and gene flow estimates based on fixation indices ( N _ST) and coalescence methods. Species samples are from the same localities along 1000 km of the Rio Juruá in western Amazonian Brazil, but each species differs in important life history traits such as population size and reproductive rate. Average haplotype differentiation, hierarchical haplotype apportionment, and gene flow estimates are contrasted in discussing the current and past population structure. Two species exhibit isolation by distance patterns wherein gene flow is largely limited to geographically adjacent localities. Mesomys exhibits this pattern throughout its range along the river. More than 75% of haplotype variation is apportioned among localities and regions, and estimates of Nm for pair-wise comparisons are nearly always less than 1. Oligoryzomys shows weak isolation by distance, but only over the largest geographical distances. Nm values for this species are nearly always above 1 and most (about 80%) of haplotype variation is contained within local populations. In contrast, Oryzomys exhibits no genetic structure throughout its entire distribution; Nm values average 17 and nearly 90% of the total haplotype variance is contained within local populations. Although gene flow estimates are high, the pattern of Nm as a function of geographical distance suggests that this species experienced a more recent invasion of the region and is still in genetic disequilibrium under its current demographic conditions.     

DNA sequencing research group: 2006 general survey of DNA sequencing facilities.

Over the past few years, technological advances in automated DNA sequencing have had a profound effect on the nature of DNA sequencing laboratories. To characterize the changes occurring within DNA sequencing facilities, the DNA Sequencing Research Group conducted three previous studies, in 1998, 2000, and 2003. A new general survey has been designed and conducted by the DSRG to capture the current status of DNA sequencing facilities in all sectors. Included were questions regarding facility administration, pricing, instrumentation, technology, protocols, and operation. The results of the survey are presented here, accompanied by comparisons to the previous surveys. These comparisons formed a basis for the discussion of trends within the facilities in response to the dynamics of a changing technology.     

Structural and functional differentiation of two clinally distributed glucosephosphate isomerase allelic isozymes from the teleost Fundulus heteroclitus

The teleost Fundulus heteroclitus (L.) possesses two loci, Gpi-A and Gpi-B, for the glycolytic enzyme, glucose-phosphate isomerase (GPI; D- glucose-6-phosphate ketol-isomerase; E.C. 5.3.1.9). The Gpi-B locus is polymorphic in Fundulus, with two common alleles, Gpi-Bb and Gpi-Bc, distributed in a clinal manner in populations along the east coast of North America. Since this clinal distribution is strongly correlated with a temperature gradient, we asked whether the GPI-B2 allozymes were functionally adapted to the thermal environment in which a given phenotype predominated. The two major GPI-B2 allozymes were purified to homogeneity and were characterized as to molecular weight, isoelectric pH, thermal denaturation, and kinetic parameters. Both GPI-Bb2 and GPI- Bc2 allozymes have molecular masses of 110 kD, and they have isoelectric pHs of 6.4 and 6.6, respectively. The GPI-Bb2 allozyme was more stable to thermal denaturation than was the GPI-Bc2 enzyme. Kinetic properties of the allelic isozymes were investigated both as a function of pH and as a function of temperature. At 25 degrees C, over the pH range considered, there were no significant differences between allozymes, either in Km for fructose-6-phosphate or in Ki for 6- phosphogluconate, but apparent Vmax values differed between pH 7.5 and pH 8.5. All steady-state kinetic parameters showed strong temperature dependence, but the allozymes differed only in the Ki for 6- phosphogluconate at temperatures greater than 30 degrees C. On the basis of the observed structural and functional differences alluded to above, the hypothesis that the major allelic isozymes of the Gpi-B locus were functionally equivalent was rejected. However, it is not yet known whether these structural and functional differences have any significance at higher levels of biological organization.      

Transpiration of trees and forest stands: short and long-term monitoring using sapflow methods

We show that sapflow is a useful tool for studies of water fluxes in forest ecosystems, because (i) it gives access to the spatial variability within a forest stand, (ii) it can be used even on steep slopes, and (iii) when combined with eddy correlation measurements over forests, it allows separation of individual tree transpiration from the total water loss of the stand. Moreover, sapflow techniques are quite easy to implement. Four sapflow techniques currently coexist, all based on heat diffusion in the xylem. We found a good agreement between three of these techniques. Most results presented here were obtained using the radial flow meter (Granier 1985). Tree sapflow is computed as sap flux density times sapwood area. To scale up from trees to a stand, measurements have to be made on a representative sample of trees. Thus, a number of trees in each circumference class is selected according to the fraction of sapwood they represent in the total sapwood area of the stand. The variability of sap flux density among trees is usually low (CV. 10–15%) in close stands of temperate coniferous or deciduous forests, but is much higher (35–50%) in a tropical rain forest. It also increases after thinning or during a dry spell. A set of 5–10 sapflow sensors usually provides an accurate estimate of stand transpiration. Transpiration measured on two dense spruce stands in the Vosges mountains (France) and one Scot's pine plantation in the Rhine valley (Germany) showed that maximum rate was related to stand LAI and to local climate. Preliminary results comparing the sapflow of a stand of Pinus banksiana to the transpiration of large branches, as part of the BOREAS programme in Saskachewan, Canada showed a similar trend. For modelling purposes, tree canopy conductance (g_c) was calculated from Penman-Monteith equation. In most experiments, calculated canopy conductance was dependent on global radiation (positive effect) and on vapour pressure deficit (negative effect) in the absence of other limiting factors. A comparison of the vapour pressure deficit response curves of g_c for several tree species and sites showed only small differences among spruce, oak and pine forests when including understorey. Tropical rainforests exhibited a similar behaviour.     

Sabellaria wilsoni sp.n. (Polychaeta, Sabellariidae) from the southeast coast of Brazil

A new species of Sabellaria is described from the southeast coast of Brazil and compared with related species known from South or North America.     

Characteristics of a thrombin inhibitor secreted by activated platelets

A 77-kDa complex of thrombin and a protein secreted by activated platelets had little if any thrombin amidolytic activity, indicating that the secreted protein is an inhibitor. The molecular weight of the inhibitor before reaction with thrombin was approximately 50,000. The apparent second-order rate constant for complex formation was estimated to be 1.3 x 10(6) M-1 s-1 (mean of four measurements); it was not affected by heparin or heparinase. These properties distinguish this inhibitor from other protease inhibitors secreted by platelets. The inhibitor reacted with trypsin and possibly with urokinase but not with factor Xa.     

A genetic and developmental analysis of an acid deoxyribonuclease in Drosophila melanogaster

A deoxyribonuclease, called D Nase-1, that is active at acid pH in the presence of EDTA has been studied in Drosophila melanogaster. The locus for the Enzyme maps genetically to 61.8 on the right arm of the third chromosome. Cytogenetically, DNase-1 has been localized to within five to ten bands between 90C-2 and 90E. This analysis utilizes both electrophoretic variants and the Y-autosome translocations of Lindsley et al. (1972). DNase-1 is present in all stages of the life cycle, and the paternal genome actively contributes DNase-1 to the ambryo between 0 and 1 hr after fertilization.     

Population genetics and conservation of critically small cycad populations: a case study of the Albany Cycad,Encephalartos latifrons (Lehmann)

Declining populations of less than 250 mature individuals are symptomatic of many Critically Endangered cycads, which, globally, comprise the most threatened group of organisms as a result of collecting and habitat loss. Survival plans focus on law enforcement, reintroduction, and augmentation programmes using plants from the wild and botanical gardens. Augmentation is one of the few remaining options for cycad populations, although the assumed benefits remain untested and there is a possibility that augmentation from different sources could compromise the genetic integrity of existing populations, especially when garden plants have no provenance data. We studied Encephalartos latifrons, a South African endemic, which is a typical Critically Endangered cycad. We studied the extent and structure of genetic diversity in wild and ex situ populations to assess the potential benefits and risks associated with augmentation programmes. We examined 86 plants using amplified fragment length polymorphisms (AFLPs). The 417 AFLP markers thus generated yielded a unique DNA ‘fingerprint’ for each plant. Wild populations retain high levels of genetic diversity and this is reflected among the ex situ holdings at the Kirstenbosch Botanical Garden. No population differentiation is evident, indicating a single panmictic population, consistent with moderately high levels of gene flow between subpopulations and a sexual mode of reproduction. Bayesian clustering identified four genotype groups in the wild, as well as a genotype group only found in ex situ collections. Our results indicate that E. latifrons would benefit from augmentation programmes, including the use of undocumented collections, and careful management of breeding plants would increase the heterogeneity of propagules. © 2011 The Linnean Society of London, Biological Journal of the Linnean Society, 2012, 105 , 293–308.