Evolution of the 28S ribosomal RNA gene in anurans: regions of variability and their phylogenetic implications

Fifteen restriction sites were mapped to the 28S ribosomal RNA gene of individuals representing 54 species of frogs, two species of salamanders, a caecilian, and a lungfish. Eight of these sites were present in all species examined, and two were found in all but one species. Alignment of these conserved restriction sites revealed, among anuran 28S rRNA genes, five regions of major length variation that correspond to four of 12 previously identified divergent domains of this gene. One of the divergent domains (DD8) consists of two regions of length variation separated by a short segment that is conserved at least throughout tetrapods. Most of the insertions, deletions, and restriction-site variations identified in the 28S gene will require sequence-level analysis for a detailed reconstruction of their history. However, an insertion in DD9 that is coextensive with frogs in the suborder Neobatrachia, a BstEII site that is limited to representatives of two leptodactylid subfamilies, and a deletion in DD10 that is found only in three ranoid genera are probably synapomorphies.      

Characterization of cross-bridge elasticity and kinetics of cross-bridge cycling during force development in single smooth muscle cells

Force development in smooth muscle, as in skeletal muscle, is believed to reflect recruitment of force-generating myosin cross-bridges. However, little is known about the events underlying cross-bridge recruitment as the muscle cell approaches peak isometric force and then enters a period of tension maintenance. In the present studies on single smooth muscle cells isolated from the toad (Bufo marinus) stomach muscularis, active muscle stiffness, calculated from the force response to small sinusoidal length changes (0.5% cell length, 250 Hz), was utilized to estimate the relative number of attached cross-bridges. By comparing stiffness during initial force development to stiffness during force redevelopment immediately after a quick release imposed at peak force, we propose that the instantaneous active stiffness of the cell reflects both a linearly elastic cross-bridge element having 1.5 times the compliance of the cross-bridge in frog skeletal muscle and a series elastic component having an exponential length-force relationship. At the onset of force development, the ratio of stiffness to force was 2.5 times greater than at peak isometric force. These data suggest that, upon activation, cross-bridges attach in at least two states (i.e., low-force-producing and high-force-producing) and redistribute to a steady state distribution at peak isometric force. The possibility that the cross-bridge cycling rate was modulated with time was also investigated by analyzing the time course of tension recovery to small, rapid step length changes (0.5% cell length in 2.5 ms) imposed during initial force development, at peak force, and after 15 s of tension maintenance. The rate of tension recovery slowed continuously throughout force development following activation and slowed further as force was maintained. Our results suggest that the kinetics of force production in smooth muscle may involve a redistribution of cross-bridge populations between two attached states and that the average cycling rate of these cross-bridges becomes slower with time during contraction.     

Synthesis and conformational studies of 2-beta-chloro, 1-alpha-fluoro, and 2-beta-fluoro derivatives of 2-deoxy-N-acetyl-neuraminic acid

5-Acetamido-4,7,8,9-tetra-O-acetyl-2,3,5-trideoxy-2-fluoro-D-glycero-alp ha- and -beta-D- galacto -2- nonulosonic acid methyl esters and the beta-chloro analog were synthesized from N-acetylneuraminic acid. Their 1H- and 13C-n.m.r.spectra were completely assigned by using single-frequency decoupling, off-resonance decoupling, and spin-simulation programs. Bond angles estimated from the 1H coupling-constants indicate that all of the compounds adopt the 2C5 (L) conformation with minor conformational differences in the C3 side chain. 5-Acetamido-2,3,5,-tri-deoxy-2-fluoro-D-glycero-alpha- and -beta-D- galacto -2- nonulosonic acid and their methyl esters were also prepared.     

Excess amino acid polymorphism in mitochondrial DNA: contrasts among genes from Drosophila, mice, and humans

Recent studies of mitochondrial DNA (mtDNA) variation in mammals and Drosophila have shown an excess of amino acid variation within species (replacement polymorphism) relative to the number of silent and replacement differences fixed between species. To examine further this pattern of nonneutral mtDNA evolution, we present sequence data for the ND3 and ND5 genes from 59 lines of Drosophila melanogaster and 29 lines of D. simulans. Of interest are the frequency spectra of silent and replacement polymorphisms, and potential variation among genes and taxa in the departures from neutral expectations. The Drosophila ND3 and ND5 data show no significant excess of replacement polymorphism using the McDonald-Kreitman test. These data are in contrast to significant departures from neutrality for the ND3 gene in mammals and other genes in Drosophila mtDNA (cytochrome b and ATPase 6). Pooled across genes, however, both Drosophila and human mtDNA show very significant excesses of amino acid polymorphism. Silent polymorphisms at ND5 show a significantly higher variance in frequency than replacement polymorphisms, and the latter show a significant skew toward low frequencies (Tajima's D = -1.954). These patterns are interpreted in light of the nearly neutral theory where mildly deleterious amino acid haplotypes are observed as ephemeral variants within species but do not contribute to divergence. The patterns of polymorphism and divergence at charge-altering amino acid sites are presented for the Drosophila ND5 gene to examine the evolution of functionally distinct mutations. Excess charge-altering polymorphism is observed at the carboxyl terminal and excess charge-altering divergence is detected at the amino terminal. While the mildly deleterious model fits as a net effect in the evolution of nonrecombining mitochondrial genomes, these data suggest that opposing evolutionary pressures may act on different regions of mitochondrial genes and genomes.      

Evidence for "twisted plane" undulations in golden hamster sperm tails

Motile spermatozoa from the golden hamster have been arrested by rapid freezing and then fixed with glutaraldehyde at low temperature after substitution with ethylene glycol. As far as can be judged, the flagellar waveforms thus stabilized are similar to those seen in living sperm; in contrast, fixation in glutaraldehyde, without prior freezing, induces agonal changes in flagellar conformation. The characteristics waveform after freeze substitution contains three bends. Approx. half of these flagella are entirely planar. The rest are three dimensional, with the third bend displaced in a regular way from the plane containing the proximal two bends. From the geometry of these flagella, it is concluded that the plane of action of a given bending cycle undergoes a clockwise twist (from a forward viewpoint) as the cycle is succeeded by new bending cycles. This "twisted plane" undulation is quite different from helical movement. The twisting seems to occur abruptly, between cycles, as if each bending cycle has a preferred plane of action. The mechanism underlying the twisting is uncertain. However, on the basis of the angular displacements between the preferred planes, and the findings from electron microscopy, the following idea is presented as a working hypothesis: that, if the most proximal plane of bending is topographically determined by peripheral doublet 1, then successive distal planes of action are influenced predominantly by doublets 2, 3, etc., in clockwise sequence. The merits and weaknesses of this hypothesis are discussed.     

The stepwise synthesis of methyl α-isomaltooligoside derivatives and methyl α-isomaltopentaoside

Methyl 2,3,4-tri-O-benzyl-α-D-glucopyranoside was treated with 2,3,4-tri-O-benzyl-6-O-(N-phenylcarbamoyl)-1-O-tosyl-D-glucopyranose in diethyl ether to give methyl 2,3,4,2',3',4'-hexa-O-benzyl-6'-O-(N-phenylcarbamoyl)-α-isomaltoside. The disaccharide was decarbanilated in ethanol with sodium ethoxide to give methyl 2,3,4,2',3',4'-hexa-O-benzyl-α-isomaltoside. The sequence of coupling with the same 1-O-tosyl-D-glucose derivative followed by removal of the N-phenylcarbamate group was repeated until the hexasaccharide derivative, methyl octadeca-O-benzyl-α-isomaltohexaoside, was formed. Methyl α-isomaltopentaoside was prepared by debenzylation of the corresponding benzylated oligosaccharide. The structures of the oligosaccharides were determined with the aid of both ¹H- and ¹³C-n.m.r. spectroscopy. From spectral data, we estimate the coupling reaction to be 95% stereoselective.     

Phenylacetaldehyde attracts male and female apple clearwing moths,Synanthedon myopaeformis,to inflorescences of showy milkweed,Asclepias speciosa

Synanthedon myopaeformis Borkhausen (Lepidoptera: Sesiidae) is a diurnal clearwing moth native to Eurasia that was recently introduced into British Columbia (BC) and Ontario, Canada, where it has become a serious pest in apple orchards. In BC, these moths commonly feed on nectar of inflorescences, particularly that of showy milkweed, Asclepias speciosa Torrey (Apocynaceae). We investigated the relative importance of visual and olfactory cues, and the key floral semiochemical(s) mediating attraction of S. myopaeformis to A. speciosa. In field experiments, inflorescences left exposed or enclosed in cheesecloth bags dyed green induced similar visitation rates by moths, indicating that olfactory cues are attractive. Among the >10 floral odourants that elicited responses from moth antennae in coupled gas chromatographic‐electroantennographic detection analyses, phenylacetaldehyde induced the most frequent proboscis extension reflexes of male and female moths. Among eight floral odourants that were field‐tested singly, phenylacetaldehyde attracted 35 times more male and female moths than any other candidate semiochemical. Attractiveness of phenylacetaldehyde could not be enhanced by admixture with other floral odourants at the ratios or concentrations tested indicating that it alone may mediate attraction of S. myopaeformis to the inflorescences of A. speciosa. The potential use of phenylacetaldehyde as bait to monitor or mass‐trap populations of male and female S. myopaeformis should be investigated.     

Deforestation Induced Climate Change: Effects of Spatial Scale

Deforestation is associated with increased atmospheric CO₂ and alterations to the surface energy and mass balances that can lead to local and global climate changes. Previous modelling studies show that the global surface air temperature (SAT) response to deforestation depends on latitude, with most simulations showing that high latitude deforestation results in cooling, low latitude deforestation causes warming and that the mid latitude response is mixed. These earlier conclusions are based on simulated large scal land cover change, with complete removal of trees from whole latitude bands. Using a global climate model we examine the effects of removing fractions of 5% to 100% of forested areas in the high, mid and low latitudes. All high latitude deforestation scenarios reduce mean global SAT, the opposite occurring for low latitude deforestation, although a decrease in SAT is simulated over low latitude deforested areas. Mid latitude SAT response is mixed. In all simulations deforested areas tend to become drier and have lower SAT, although soil temperatures increase over deforested mid and low latitude grid cells. For high latitude deforestation fractions of 45% and above, larger net primary productivity, in conjunction with colder and drier conditions after deforestation cause an increase in soil carbon large enough to produce a net decrease of atmospheric CO₂. Our results reveal the complex interactions between soil carbon dynamics and other climate subsystems in the energy partition responses to land cover change.     

Models of Eucalypt phenology predict bat population flux

Fruit bats (Pteropodidae) have received increased attention after the recent emergence of notable viral pathogens of bat origin. Their vagility hinders data collection on abundance and distribution, which constrains modeling efforts and our understanding of bat ecology, viral dynamics, and spillover. We addressed this knowledge gap with models and data on the occurrence and abundance of nectarivorous fruit bat populations at 3 day roosts in southeast Queensland. We used environmental drivers of nectar production as predictors and explored relationships between bat abundance and virus spillover. Specifically, we developed several novel modeling tools motivated by complexities of fruit bat foraging ecology, including: (1) a dataset of spatial variables comprising Eucalypt‐focused vegetation indices, cumulative precipitation, and temperature anomaly; (2) an algorithm that associated bat population response with spatial covariates in a spatially and temporally relevant way given our current understanding of bat foraging behavior; and (3) a thorough statistical learning approach to finding optimal covariate combinations. We identified covariates that classify fruit bat occupancy at each of our three study roosts with 86–93% accuracy. Negative binomial models explained 43–53% of the variation in observed abundance across roosts. Our models suggest that spatiotemporal heterogeneity in Eucalypt‐based food resources could drive at least 50% of bat population behavior at the landscape scale. We found that 13 spillover events were observed within the foraging range of our study roosts, and they occurred during times when models predicted low population abundance. Our results suggest that, in southeast Queensland, spillover may not be driven by large aggregations of fruit bats attracted by nectar‐based resources, but rather by behavior of smaller resident subpopulations. Our models and data integrated remote sensing and statistical learning to make inferences on bat ecology and disease dynamics. This work provides a foundation for further studies on landscape‐scale population movement and spatiotemporal disease dynamics.     

Alternative hypotheses for mammalian herbivore preference of burned areas in a savannah ecosystem

Mammalian herbivores prefer burned areas and this attraction has largely been attributed to the increased nutrient content of the postfire green flush and more recently to the avoidance of predators. However, alternative reasons for this attraction could be: (i) to avoid disease carrying and behaviour changing invertebrates; (ii) because burned areas are warmer microclimates; or (iii) to obtain minerals from the ash. This study tests for differences in tick and fly (Diptera) numbers between burned and unburned areas in Serengeti National Park, Tanzania. It also tests for differences in ground and air column temperatures between burned and unburned areas and for differences in the mineral content of ash in burned areas compared to the mineral content of green leaves in unburned areas. We found no difference in the abundance of either type of invertebrate between burned and unburned areas. Only ground temperature was higher in burned areas and this was only during the middle of the day, when increases in temperature would be less important than at night. Ash was higher in Al, Ca, Cu, Mg, Mn and P than nearby green leaves from unburned vegetation. Thus, obtaining minerals from ash is the only alternative reason for attraction to burned areas that maybe supported by this study.