Novobiocin-Induced Ultrastructural Changes and Antagonism of DNA Synthesis in Trypanosoma cruzi Amastigotes Growing in Cell-Free Medium

The antimicrobial agent novobiocin, an inhibitor of the bacterial enzyme topoisomerase II (DNA gyrase), is known to antagonize Trypanosoma cruzi amastigotes growing in cell-free medium. To determine sites of antagonism of novobiocin, the effects of drug on parasite ultrastructure and incorporation of radiolabeled precursors of DNA, RNA and protein into macromolecules were determined. The predominant ultrastructural abnormality seen after exposure to 0.40 mM novobiocin for 24 h was the presence of electron-dense clumps in the mitochondrion-kinetoplast organelle in 95 of 257 (37%) of cells, in comparison to no clumps seen in 110 drug-free cells. In addition, in the nucleus, the karyosome was less distinct than in control cells and appeared to merge with the chromatin. In the radiolabcling studies, incorporation of thymidine was inhibited in a dose-dependent fashion by novobiocin (0.16–0.80 mM) in a range of drug concentrations that also inhibited parasite growth. For 0.16 and 0.24 mM novobiocin, incorporation of thymidine was inhibited up to 65% relative to drug-free control cells while uptake of uridine and leucine was unaltered. We interpret these ultrastructure and precursor-incorporation studies as suggesting that (i) the mitochondrion-kinetoplast and possibly the nucleus are sites of novobiocin antagonism of T. cruzi amastigotes and (ii) that novobiocin appears to antagonize DNA synthesis within these organisms. Whether the drug target is topoisomerase II, however, is as yet unknown.     

Slow continuous wing-moult of Zebra Finches Poephila guttata from southeast Australia

Moult of the remiges was studied over eight years in a population of Zebra Finches from southeast Australia. In first-year birds the first primary was lost at mean age of 80 days in both sexes; moult took 204 days to complete in males and 223 in females. Birds hatched in autumn postponed the start of their primary moult until spring. Primaries were replaced slowly in a rigid, non-overlapping, ascending order from #1 to #9 but the secondaries did not display a rigid sequence.
Adult Zebra Finches moulted remiges throughout the year. Primary moult was initiated in any month. Primary moult had a mean duration of 229 days for males and 240 days for females. Primaries moulted in continuous waves: the interval between successive moults in males was –16 days and +6 days in females. Limited food, low temperatures and breeding slowed, but did not stop, wing-moult. Birds caring for eggs and young actively replaced remiges.
The pattern of wing-moult displayed here by Zebra Finches is exceptional for passerines and may be related to the opportunistic breeding strategy necessary in an arid, unpredictable environment. Its presence in the Victorian birds may be non-adaptive.     

Migration of alate morphs of the bird cherry aphid (Rhopalosiphum padi) and implications for the epidemiology of barley yellow dwarf virus

Suction trapping data indicate three periods of migration of Rhopalosiphum padi in spring, summer and autumn. Four alate morphs are present at different times during the year. A comparison of data from suction traps operating at 12·2 and 1·5 m suggests a different behaviour of females in autumn with more being recorded at 12·2 than 1·5 m. Males, which are only present in autumn, were also more numerous at 12·2 m. During tests to measure barley yellow dwarf virus (BYDV) infectivity, only 9% of female R. padi reproduced on oat seedlings in autumn compared with 74% in summer. Tests on alate female R. padi trapped alive showed that in summer all were exules, but during the first half of September these were largely replaced by gynoparae so that in autumn only 5% of all R. padi trapped at 12·2 m were alate exules. The aerial densities of gynoparae and males were 10 times greater at 12·2 than 1·5 m while densities of alate exules were similar at both heights. It is suggested that gynoparae and males fly higher to increase the chance of finding a taller dispersed host plant. The implications for BYDV epidemiology of the behaviour and presence of the various R. padi alate morphs indicate that autumn-sown cereals emerging before mid-September are particularly at risk from colonisation by alate exules before the transition to a mainly sexual migrant population is complete. Alate exules introduce BYDV from comparatively local sources. The ratio of total R. padi to Sitobion avenae in suction trap samples in autumn usually exceeds 100: 1, but on crops it was only 10: 1. The ratio of alate exule R. padi to S. avenae in suction traps in autumn was only 12: 1, similar to that observed on crops.     

AN EMPIRICAL COMPARISON OF NUMERICAL WAGNER COMPUTER PROGRAMS

Abstract— The performance of four computer programs that calculate Wagner trees (WAGNER 78, WAGPROC, PHYLIP, and PHYSYS) was compared for twenty-five data sets. Eight combinations of algorithms and options were tried, including different methods of adding taxa, optimizing stem states, obtaining multiple trees, and branch swapping. Using the criterion of finding a minimum length tree, PHYSYS with the WAG.S option performed best, providing the shortest tree for twenty-four of the twenty-five data sets. WAGPROC with the GLOB option found sixteen minima for eighteen data sets, exceeding run time on the remaining seven. All other algorithm/options were less successful in providing minimum trees. In comparing the options we found that minimum homoplasy is not completely reliable in optimizing trees and that the brute force algorithm is helpful but not required for finding minimum trees. The advancement index criterion for adding taxa to a tree is more effective than adding taxa in their data file sequence. The success of the PHYSYS WAG.S option and the WAGPROC GLOB demonstrate that both multiple trees and branch swapping are necessary to produce a minimum length tree.     

The cytotoxic and photodynamic inactivation of micro-organisms by Rose Bengal

Rose Bengal was cytotoxic to the following bacteria at the concentrations given in parentheses (highest concentrations of dye in mol/1 at which growth occurred on nutrient medium): Brochothrix thermosphacta and Deinococcus radiodurans (1 times 10^-6 or less); Streptococcus, Micrococcus, Staphylococcus, Bacillus, Arthrobacter and Kurthia spp. (1 times 10^-5–1 x 10^-4), and Pseudomonas spp. and Enterobacteriaceae (5 times 10^-3–1 x 10^-2 or greater). These organisms were killed rapidly when suspended in illuminated (170 μE/m²/s) solutions of Rose Bengal (1 times 10^-4 mol/1) providing oxygen was present. Singlet oxygen was identified as the lethal agent, because the rate of killing was increased by dissolving the dye in deuterium oxide while the organisms were protected against photoinactivation by L-histidine or crocetin. Yeasts from chilled foods were killed in illuminated solutions of Rose Bengal but a light intensity of 315 μE/m²/s was needed for a death rate comparable with that of bacteria. The yeasts present in a range of chilled meat and dairy products failed to form colonies on Rose Bengal (5 times 10^-5 mol/1) media exposed continuously to modest illumination (55–80 μE/m²/s).     

Alternate Bearing Affects Nitrogen, Phosphorus, Potassium and Starch Storage Pools in Mature Pistachio Trees

The relationship between crop load and the functional storageof selected macronutrients and starch was assessed to developnutrient budgets and best management fertilization practicesin orchards. Functional storage represents the amount of nutrientsand starch redistributed from perennial tree parts in supportof the spring growth flush. Functional storage was influencedby:(a)nutrient and starch accumulation prior to dormancy; and(b)nutrientand starch demand by vegetative and reproductive organs in spring.Lightly cropping (off-year) trees stored 7, 14 and 2 times asmuch N, P and K, respectively, as heavily cropping (on-year)trees. Similar to many biennial plant species, nutrients thataccumulated during the vegetative phase in off-year trees wereused to support reproductive growth during the subsequent on-year.Soil nutrient uptake contributed more to storage pools thanleaf nutrient resorption in off-year-trees, while the reversewas true in on-year trees. Net nutrient resorption from senescingleaves accounted for all of the N and P and a third of the Kstored in on-year trees. Only between 20–33% of the N,P and K stored in perennial tissues of off-year trees couldbe attributed to leaf nutrient resorption. This is the firststudy to determine the amounts of nutrients stored in the perennialparts of mature, field-grown trees and the relative contributionsof leaf nutrient resorption and soil nutrient uptake to functionalstorage in trees.Copyright 1998 Annals of Botany Company Pistacia vera, nutrient storage, biennial bearing, crop load, leaf nutrient resorption, source-sink relationships.     

Epigenetic mechanical factors in the evolution of long bone epiphyses

In developing vertebrate long bones in which endochondral ossification occurs, it is preceded or accompanied by perichondral ossification. The speed and extent of perichondral apposition relative to endochondral ossification varies in different taxa. Perichondral ossification dominates early long bone development in extinct basal tetrapods and dinosaurs, extant bony fish, amphibians, and birds. In mammals and lizards, perichondral and endochondral ossification proceed more synchronously. One of the most important epigenetic factors in skeletogenesis is mechanical loading caused by muscle contractions which begin in utero or in ovo . It has been previously shown that the stress distributions created perinatally in the chondroepiphysis during human skeletal development can influence the appearance of secondary ossification centres. Using finite element computer models representing bones near birth or hatching, we demonstrate that in vertebrates in which perichondral ossification significantly precedes endochondral ossification, the distribution of mechanical stresses in the ossifying cartilage anlagen tends to inhibit the appearance of secondary ossification centres in the ends of long bones. In models representing vertebrates in which endochondral ossification keeps pace with perichondral apposition, the appearance of secondary centres is promoted. The appearance of secondary centres leads to the formation of bony epiphyses and growth plates, which are most common in mammals and extant lizards. We postulate that genotypic factors influencing the relative speed and extent of perichondral and endochondral ossification interact with mechanical epigenetic factors early in development to account for many of the morphological differences observed in vertebrate skeletons.     

Mitochondrial DNA variation in natural populations of the mushroom Agaricus bisporus

We investigated the patterns of mitochondrial DNA variation in the global population of the commercial mushroom Agaricus bisporus . Through the analysis of RFLP's among 441 isolates from nine countries in North America and Eurasia, we found a total of 140 mtDNA haplotypes. Based on population genetic analysis, there are four genetically distinct natural populations in this species, found in coastal California, desert California, France and Alberta (Canada). While 134 of the 140 mtDNA haplotypes were unique to single geographical regions, two mtDNA haplotypes, mt001 and mt002, were found in almost every population surveyed. These two mtDNA haplotypes also predominate among cultivars used throughout the world for at least the last two decades. These two mtDNA haplotypes are more similar to the cosmopolitan groups of mtDNA haplotypes than to the indigeneous clusters of mtDNA haplotypes from the two Californian regions.     

Consequences of habitat loss and change to populations of wintering migratory birds: predicting the local and global effects from studies of individuals

Population models show that the response of migratory bird populations to a reduction in the area, or change in the quality, of their winter feeding areas depends critically on the shape (linear or non-linear) and strength (slope) of the relationship describing how the proportion dying from starvation changes with population density. Yet the parameters that define such density-dependent functions are extremely difficult to estimate from direct measurements of mortality at different population sizes. Even if they could be estimated, there would be uncertainty as to whether or not the relationship would remain the same, and thus provide reliable predictions, in the new circumstances for which forecasts are required. This paper summarizes studies of the Oystercatcher Haematopus ostralegus which aim to derive the shape and strength of the winter density-dependent mortality and emigration function for one estuary, under existing and new circumstances, from the responses of individual birds to each other and to their spatially and seasonally varying food supply. Based on these studies, an individuals-based, physiologically structured game theoretic distribution model has been built which predicts the carrying capacity and numbers of birds dying at different population sizes. The model also can be applied to Oyster-catchers occupying several estuaries distributed throughout their wintering range and can thus be extended to the entire biogeographical or global population. In addition, it can be used to identify some easily measured behavioural and ecological parameters that identify those species, from a wide range of taxa, that are most likely to be affected by habitat loss and change.     

Two New Menosporine Gregarines, Hoplorhynchus acanthatholius N. Sp. and Steganorhynchus dunwoodyi N. G., N. Sp. (Apicomplexa: Eugregarinorida: Actinocephalidae) from Coenagrionid Damselflies (Odonata: Zygoptera)

ABSTRACT. Hoplorhynchus acanthatholius , n. sp. is described from Enallagma civile , the Civil Bluet damselfly. Trophozoites are solitary, lie in the mesenteron between the peritrophic membrane and the epithelium, and attain a maximum length of 850 μm. Epimerite ovoid to broadly ovoid; anterior margin bearing eight equidistant retroarcuate hooks; attached to protomerite by means of a vermicular stalk. Protomerite ovoid; deutomerite narrowly obvoid. Gametocysts spherical; diam 300 μm, sporulating by simple dehiscence in 48–72 h. Oocysts are characteristic of Menosporinae: smooth, biconical, crcscentic, uniform in size and shape. Steganorhynchus dunwoodyi , n. g., n. sp. is described from the damselfly Ischnura verticalis. The genus is characterized by an epimerite comprising an ovoid papilla enclosed in a retractable, globular sheath, borne on a long vermicular stalk. Trophozoites are solitary, lie in the mesenteron between the peritrophic membrane and epithelium, and attain a maximum length of 605 μm. Protomerite very broadly ovoid; deutomerite obvoid. Gametocysts spherical; diam 258 μm, sporulating by simple dehiscence in 48–72 h. Oocysts are characteristic of Menosporinae: smooth, biconical, crescentic, uniform in size and shape. The population dynamics of H. acanthatholius and S. dunwoodyi among damselfly populations in five Nebraska localities are presented.