Effects of Roads on Endangered San Joaquin Kit Foxes

ABSTRACT San Joaquin kit foxes (Vulpes macrotis mutica) occur in central California, USA, and are endangered due to habitat loss and degradation. As the human population of California grows, more roads are being constructed in remaining kit fox habitat. We examined effects of 2-lane roads on demographic and ecological patterns of kit foxes on the Lokern Natural Area (LNA) from August 2001 to June 2004. Of 60 radiocollared kit foxes, only one was struck by a vehicle. Foxes were assigned to 1 of 3 risk categories (high, medium, or low) based on proportion of time spent in road-effect zones, which were defined by the probability of a fox encountering a road during nocturnal movements. Fox survival probabilities, reproductive success, litter size, nocturnal movements, and den placement all were similar among risk categories. Nocturnal locations of foxes were closer to roads than were den locations, and den fidelity was lowest for medium-risk foxes and highest for low-risk foxes but intermediate for high-risk foxes. Food availability and use were not affected by proximity to roads. We were unable to detect any significant detrimental effects from 2-lane roads on kit fox demography and ecology. Our results suggest that standard mitigation strategies, such as crossing structures and exclusionary fencing, would not benefit kit foxes on the LNA.     

Cross-hybridizing snake satellite, Drosophila, and mouse DNA sequences may have arisen independently

Previous reports have interpreted hybridization between snake satellite DNA and DNA clones from a variety of distant taxonomic groups as evidence for evolutionary conservation, which implies common ancestry (homology) and/or convergence (analogy) to produce the cross- hybridizing sequences. We have isolated 11 clones from a genomic library of Drosophila melanogaster, using a cloned 2.5-kb snake satellite probe of known nucleotide sequence. We have also analysed published sequence data from snakes, mice, and Drosophila. These data show that (1) all of the cross-hybridization between the snake, fly, and mouse clones can be accounted for by the presence of either of two tandem repeats, [GATA]n and [GACA]n and (2) these tandem repeats are organized differently among the different species. We find no evidence that these sequences are homologous apart from the existence of the simple repeat itself, although their divergence from a common ancestral sequence cannot be ruled out. The sequences contain a variety of homogeneous clusters of tandem repeats of CATA, GA, TA, and CA, as well as GATA and GACA. We suggest that these motifs may have arisen by a self-accelerating process involving slipped-strand mispairing of DNA. Homogeneity of the clusters might simply be the result of a rate of accumulation of tandem repeats that exceeds that of other mutations.      

Structural comparison of fibroblast growth factor-specific heparan sulfates derived from a growing or differentiating neuroepithelial cell line

Heparan sulfate (HS) glycosaminoglycans are essential modulators of fibroblast growth factor (FGF) activity both in vivo and in vitro, and appear to act by cross-linking particular forms of FGF to appropriate FGF receptors. We have recently isolated and characterized two separate HS pools derived from immortalized embryonic day 10 mouse neuroepithelial 2.3D cells: one from cells in log growth phase, which greatly potentiates the activity of FGF-2, and the other from cells undergoing contact-inhibition and differentiation, which preferentially activates FGF-1. These two pools of HS have very similar functional activities to those species isolated from primary neuroepithelial cells at corresponding stages of active proliferation or differentiation. We present here a structural comparison between these cell line HS species to establish the nature of the changes that occur in the biosynthesis of HS. A combination of chemical and enzymatic cleavage, low pressure chromatography and strong anion-exchange HPLC were used to generate full chain models of each species. Overall, the HS pools synthesized in the dividing cell line pools possessed less complex sulfation than those derived from more differentiated, growth arrested cells.      

Effects of nucleotide sequence alignment on phylogeny estimation: a case study of 18S rDNAs of apicomplexa

The reconstruction of phylogenetic history is predicated on being able to accurately establish hypotheses of character homology, which involves sequence alignment for studies based on molecular sequence data. In an empirical study investigating nucleotide sequence alignment, we inferred phylogenetic trees for 43 species of the Apicomplexa and 3 of Dinozoa based on complete small-subunit rDNA sequences, using six different multiple-alignment procedures: manual alignment based on the secondary structure of the 18S rRNA molecule, and automated similarity-based alignment algorithms using the PileUp, ClustalW, TreeAlign, MALIGN, and SAM computer programs. Trees were constructed using neighboring-joining, weighted-parsimony, and maximum- likelihood methods. All of the multiple sequence alignment procedures yielded the same basic structure for the estimate of the phylogenetic relationship among the taxa, which presumably represents the underlying phylogenetic signal. However, the placement of many of the taxa was sensitive to the alignment procedure used; and the different alignments produced trees that were on average more dissimilar from each other than did the different tree-building methods used. The multiple alignments from the different procedures varied greatly in length, but aligned sequence length was not a good predictor of the similarity of the resulting phylogenetic trees. We also systematically varied the gap weights (the relative cost of inserting a new gap into a sequence or extending an already-existing gap) for the ClustalW program, and this produced alignments that were at least as different from each other as those produced by the different alignment algorithms. Furthermore, there was no combination of gap weights that produced the same tree as that from the structure alignment, in spite of the fact that many of the alignments were similar in length to the structure alignment. We also investigated the phylogenetic information content of the helical and nonhelical regions of the rDNA, and conclude that the helical regions are the most informative. We therefore conclude that many of the literature disagreements concerning the phylogeny of the Apicomplexa are probably based on differences in sequence alignment strategies rather than differences in data or tree-building methods.      

Leaf gas exchange and nitrogen dynamics of N2-fixing, field-grown Alnus glutinosa under elevated atmospheric CO2

Few studies have investigated the effects of elevated CO₂ on the physiology of symbiotic N₂-fixing trees. Tree species grown in low N soils at elevated CO₂ generally show a decline in photosynthetic capacity over time relative to ambient CO₂ controls. This negative adjustment may be due to a reallocation of leaf N away from the photosynthetic apparatus, allowing for more efficient use of limiting N. We investigated the effect of twice ambient CO₂ on net CO₂ assimilation (A), photosynthetic capacity, leaf dark respiration, and leaf N content of N2-fixing Alnus glutinosa (black alder) grown in field open top chambers in a low N soil for 160 d. At growth CO₂, A was always greater in elevated compared to ambient CO₂ plants. Late season A vs. internal leaf p(CO₂) response curves indicated no negative adjustment of photosynthesis in elevated CO₂ plants. Rather, elevated CO₂ plants had 16% greater maximum rate of CO₂ fixation by Rubisco. Leaf dark respiration was greater at elevated CO₂ on an area basis, but unaffected by CO₂ on a mass or N basis. In elevated CO₂ plants, leaf N content (μg N cm^?2) increased 50% between Julian Date 208 and 264. Leaf N content showed little seasonal change in ambient CO₂ plants. A single point acetylene reduction assay of detached, nodulated root segments indicated a 46% increase in specific nitrogenase activity in elevated compared to ambient CO₂ plants. Our results suggest that N₂-fixing trees will be able to maintain high A with minimal negative adjustment of photosynthetic capacity following prolonged exposure to elevated CO₂ on N-poor soils.     

Selection for and against insecticide resistance and possible methods of inhibiting the evolution of resistance in mosquitoes

Abstract. 1. From published data on the rate of build-up of insecticide resistance in wild populations of Anopheles culicifacies and An.stephensi , calculations have been made of the coefficients of selection which would be required to produce these rates of evolution. It is assumed in turn that the resistances might be effectively dominant, recessive or intermediate.
2. The results in the case of DDT resistance in An.culicifacies indicated that the fitness of resistant individuals was 1.28–1.51 × that of susceptibles.
3. Published data have also been examined with reference to the reversions towards susceptibility which occurred when spraying has been withdrawn. The coefficients of selection against resistance, which would be required to produce the observed rates of reversion, have been calculated. In the case of An.culicifacies and DDT these coefficients were found to be inversely related to the initial frequency of the susceptibility gene.
4. The coefficients of selection have been used to assess the feasibility of inhibiting the evolution of resistance by spraying different insecticides in the alternate sectors of a grid pattern and/or periodically releasing susceptible males. If the proportion of migrants from sector to sector were 15–30%, a "grid" system would considerably reduce the rate of evolution of resistance, but to prevent resistance from ultimately building up to high levels periodic releases would also be required. These releases need not be frequent (approximately during one month each year) if resistance is effectively recessive in the field.     

Inhibition of Reproduction in <Emphasis Type="Italic">Trypanosoma lewisi</Emphasis> by Ouabain

Trypanosoma lewisi usually produces a benign parasitaemia in rats. The parasites multiply only during the first few days after infection. Reproducing populations contain organisms that vary greatly in shape, size and total length. The coefficient of variation (CV) of the total length of the parasites measures this variability and is a sensitive index of reproductive activity¹. The CV is high, about 25%, early in the infection when reproduction is intense and thereafter decreases during infection until the trypanosomes become nearly uniform in size at about the tenth day of infection after which mostly non-reproducing forms are found².     

Repair of DNA-protein cross-links in mammalian cells

DNA-protein cross-links are generated by both endogenous and exogenous DNA damaging agents, as intermediates during normal DNA metabolism, and during abortive base excision repair. Cross-links are relatively common lesions that are lethal when they block progression of DNA polymerases. DNA-protein cross-links may be broadly categorized into four groups by the DNA and protein chemistries near the cross-link and by the source of the cross-link: DNA-protein cross-links may be found (1) in nicked DNA at the 3' end of one strand (topo I), (2) in nicked DNA at the 5' end of one strand (pol beta), (3) at the 5' ends of both strands adjacent to nicks in close proximity (topo II; Spo 11), and (4) in one strand of duplex DNA (UV irradiation; bifunctional carcinogens and chemotherapeutic agents). Repair mechanisms are reasonably well-defined for groups 1 and 3, and suggested for groups 2 and 4. Our work is focused on the recognition and removal of DNA-protein cross-links in duplex DNA (group 4).     

Are there effective resistance management strategies for vectors of human disease?*

The rotation of insecticides used by the Onchocerciasis Control Programme in West Africa is reviewed and the motives for this rotation are shown to be not only management of temephos resistance in the Simulium vectors but also constraints on what compounds are usable at particular seasons. A computer model indicates that without these seasonal constraints there is unlikely to be an advantage in a pre-planned rotation of insecticides, as compared with the prompt switching of compounds as dictated by detection of build up of resistance and switching back to the original compound if the regression of resistance is found to give the opportunity to do so. The latter sequence of events can hardly be called a ‘strategy’ for resistance management, but is what any well-managed pest control programme would be expected to do. The use of an insecticide mixture is different in principle from the use of rotation and depends on the idea that if the mixture is used from the outset, when resistance to both components of the mixture is likely to be rare, the double resistance combination would be so rare as to be dwarfed in numbers by those insects which avoid exposure altogether. The prospects for successful use of a mixture depend on each component killing a very high percentage of the exposed insects which are genetically susceptible to it. Whether this condition is met could be tested, for example, in the case of exposure of mosquitoes to insecticide-treated bednets.