Effect of the anticoccidial arprinocid on production, sporulation, and infectivity of Eimeria oocysts

Medication of broilers with arprinocid [MK-302, 9-(2-chloro-6-fluorbenzyl adenine)] had 3 distinct effects on oocysts; (1) the number of oocysts produced was decreased, (2) fewer of the oocysts sporulated, and (3) those oocysts which did sporulate were less infective than those from unmedicated birds. The drug level necessary to prevent passage of oocysts depended on the species and strain of coccidia. To essentially eliminate oocyst production (less than 5% of controls) required medication with the following levels of arprinocid: 70 ppm with Eimeria maxima; 60 ppm with E. mivati, E. E. necatrix, and E. brunetti; and 50 ppm with E. tenella. With E. acervulina, oocysts were completely eliminated by 60 ppm of arprinocid with one field strain but were still numerous at 70 ppm with a second field strain. Oocysts recovered from birds on medication often failed to sporulate. No sporulation was seen at drug levels of 30 ppm or above with E. maxima and E. mivati. The level of arpinocid required to prevent sporulation with other species depended on the strain being studied, but varied from 30 ppm to 70 ppm. The oocysts of E. acervulina, E. mivati, E. tenella, and E. brunetti recovered from medicated birds that subsequently sporulated, were less infective when inoculated into susceptible birds, than oocysts from unmedicated birds. Oocysts from low medication level with E. necatrix (30 ppm) and E. maxima (10 ppm), once sporulated, were as infective as oocysts from unmedicated control birds, even though the numbers produced were less. No differences were detected in the time oocysts were produced between medicated and unmedicated birds infected with E. acervulina, E. maxima, E. brunetti, and E. tenella.     

Plants in the forest canopy: some reflections on current research and future direction

Plants are one of the sessile components of the forest canopy, and consequently quantitative studies of plant components are more widely available than for some of their mobile counterparts. From a global perspective, several exciting and innovative canopy access tools have been designed over the past few decades that have facilitated the expansion our understanding of canopy plants. These include a network of canopy cranes, the development of the French luge (or sled), and the construction of walkways and platforms for access into different levels of the canopy on a permanent basis. At the recent international forest canopy conference in Sarasota, Florida (Forest Canopies 1998: Global Perspectives, November 4–8, 1998), many canopy plant studies were presented that illustrated the achievements in this field. As co-chair at this event, I present here some reflections based on my observations of the development of canopy research between the first and second international conferences. A selection of case studies from the conference presentations are cited in this review, including: tree crown respiration studies from the crane in Panama; epiphyte ecology from walkways in Peru; insect-plant relationships in tree crowns of the USA using platforms; experimental studies of epiphytes in cloud forests using single rope techniques; and epiphyte diversity surveys in Africa. Ideas for the future are also mentioned such as the novel concept of canopy farming of orchids in Costa Rica, the linking of canopy processes to forest floor activities, and the construction of canopy walkways to provide a sustainable forest economy instead of the conventional practice of logging. The integration of research with sustainable use of forests provides a conservation theme for future canopy studies. Such new approaches to studies of canopy plants are important, as scientists increasingly play a role in global conservation policies.     

Down-regulation of Pkc1-mediated signaling by the deubiquitinating enzyme Ubp3

Regulated ubiquitination and degradation of signaling proteins have emerged as key mechanisms for modulating the strength and duration of signaling pathways. The reversible nature of the ubiquitination process as well as the large number and diversity of the deubiquitinating enzymes raise the possibility that signaling pathways might be modulated by specific deubiquitinating enzyme(s). Here we provide evidence that in the yeast Saccharomyces cerevisiae, the Pkc1-mediated signaling pathway that controls the cell wall integrity is negatively regulated by the deubiquitinating enzyme Ubp3. Disruption of the UBP3 gene leads to an enhanced activation of the cell wall integrity pathway MAPK Slt2 when cells are challenged with a variety of pathway activation agents such as pheromone and Congo red. The ubp3 deletion mutants accumulate high levels of Pkc1, suggesting potential regulation of Pkc1 by Ubp3. Consistent with this, Pkc1 and Ubp3 interact in vivo, and the stability of Pkc1 is markedly increased in the ubp3 deletion mutants. Moreover, disruption of the PKC1 gene, but not the genes that encode components downstream of Pkc1, completely suppresses other phenotypes displayed by the ubp3 deletion mutants such as hyperactivation of the pheromone-responsive MAPK Fus3 (Wang, Y., and Dohlman, H. G. (2002) J. Biol. Chem. 277, 15766-15772). These findings demonstrate that Ubp3 can regulate Pkc1 by facilitating its destruction and provide the initial evidence that Pkc1 plays a positive role in modulating the parallel pheromone-signaling pathway.     

The institutional official and postapproval monitoring: the view from 10,000 feet

The role of the institutional official (IO) in ensuring a high-quality program of laboratory animal care and use has received relatively little attention in the literature compared to that of the institutional animal care and use committee (IACUC) or attending veterinarian. Yet the IO is critical to institutional effectiveness. Often not a scientist, especially at colleges and universities, the IO is an executive who bears ultimate responsibility for the care and use of laboratory animals. An IO operates largely through delegation to the IACUC and the institutional veterinarian, yet must maintain a clear view of the program from the executive level--the "view from 10,000 feet." To do so, the IO must bring four critical leadership qualities to the position: (1) vision, (2) a commitment to quality and integrity, (3) strong planning and resource development, and (4) accountability to the laboratory animal program. The IO's ability to act on these qualities and to evaluate the effectiveness of the resulting program requires reliance on a strong program of postapproval monitoring (PAM). Summary data from PAM, based on metrics carefully defined by the IO in consultation with the IACUC and the institutional veterinarian, provide the feedback necessary to guide executive decisions and allow the IO to place the needs of the program of laboratory animal care and use successfully into the larger context of the mission of the institution.     

Canine size, shape, and bending strength in primates and carnivores

Anthropoid primates are well known for their highly sexually dimorphic canine teeth, with males possessing canines that are up to 400% taller than those of females. Primate canine dimorphism has been extensively documented, with a consensus that large male primate canines serve as weapons for intrasexual competition, and some evidence that large female canines in some species may likewise function as weapons. However, apart from speculation that very tall male canines may be relatively weak and that seed predators have strong canines, the functional significance of primate canine shape has not been explored. Because carnivore canine shape and size are associated with killing style, this group provides a useful comparative baseline for primates. We evaluate primate maxillary canine tooth size, shape and relative bending strength against body size, skull size, and behavioral and demographic measures of male competition and sexual selection, and compare them to those of carnivores. We demonstrate that, relative to skull length and body mass, primate male canines are on average as large as or larger than those of similar sized carnivores. The range of primate female canine sizes embraces that of carnivores. Male and female primate canines are generally as strong as or stronger than those of carnivores. Although we find that seed-eating primates have relatively strong canines, we find no clear relationship between male primate canine strength and demographic or behavioral estimates of male competition or sexual selection, in spite of a strong relationship between these measures and canine crown height. This suggests either that most primate canines are selected to be very strong regardless of variation in behavior, or that primate canine shape is inherently strong enough to accommodate changes in crown height without compromising canine function.     

No Evidence for Kin Discrimination in Cannibalistic Tree-Hole Mosquitoes (Diptera: Culicidae)

Kin selection theory suggests that cannibalism is more likely to spread and be maintained if cannibalism of close relatives can be preferentially avoided. One important group of insects in which kin discrimination might be expected to evolve is cannibalistic tree-hole mosquitoes. Larvae of these species develop in small, ephemeral water bodies, where they regularly encounter both relatives and nonrelatives. In this study we compared the degree of sib cannibalism with the degree of nonsib cannibalism in two ecologically distinct mosquito species: an anautogenous (blood-feeding) detritivore Trichoprosopon digitatum (Culicinae) and an autogenous predator Toxorhynchites moctezuma (Toxorhynchitinae). Despite a wealth of literature documenting kin-biased discrimination in other insects, neither of these tree-hold mosquito species preferentially consumed nonrelatives. In the case of Tr. digitatum, the size-dependent nature of larval cannibalism may preclude additional selection for preferential consumption of nonrelatives, but in the autogenous Tx. moctezuma the direct nutritive and indirect competitive benefits of indiscriminate cannibalism may outweigh the immediate costs of consuming a relative.     

The anomalous archaic Homo femur from Berg Aukas, Namibia: a biomechanical assessment.

The probably Middle Pleistocene human femur from Berg Aukas, Namibia, when oriented anatomically and analyzed biomechanically, presents an unusual combination of morphological features compared to other Pleistocene Homo femora. Its midshaft diaphyseal shape is similar to most other archaic Homo, but its subtrochanteric shape aligns it most closely with earlier equatorial Homo femora. It has an unusually low neck shaft angle. Its relative femoral head size is matched only by Neandertals with stocky hyperarctic body proportions. Its diaphyseal robusticity is modest for a Neandertal, but reasonable compared to equatorial archaic Homo femora. Its gluteal tuberosity is relatively small. Given its derivation from a warm climatic region, it is best interpreted as having had relatively linear body proportions (affecting proximal diaphyseal proportions, shaft robusticity, and gluteal tuberosity size) combined with an elevated level of lower limb loading during development (affecting femoral head size and neck shaft angle).     

Redesigning an antibody fragment for faster association with its antigen

Traditional approaches for increasing the affinity of protein-protein complexes focus on constructing highly complementary binding surfaces. Recent theoretical simulations and experimental results suggest that electrostatic steering forces can also be manipulated to increase association rates while leaving dissociation rates unchanged, thus increasing affinity. Here we demonstrate that electrostatic attraction can be enhanced between an antibody fragment and its cognate antigen through application of a few simple rules to identify potential on-rate amplification sites that lie at the periphery of the antigen-antibody interface.     

Long bone articular and diaphyseal structure in Old World monkeys and apes. II: Estimation of body mass

Body mass estimation equations are generated from long bone cross-sectional diaphyseal and articular surface dimensions in 176 individuals and 12 species of hominoids and cercopithecoids. A series of comparisons is carried out to determine the best body mass predictors for each of several taxonomic/locomotor groupings. Articular breadths are better predictors than articular surface areas, while cross-sectional shaft strengths are better predictors than shaft external breadths. Percent standard errors of estimate (%SEEs) and percent prediction errors for most of the better predictors range between 10-20%. Confidence intervals of equations using sex/species means are fairly representative of those calculated using individual data, except for sex/species means equations with very low %SEEs (under about 10%), where confidence intervals (CIs) based on individuals are likely to be larger. Given individual variability, or biological "error," this may represent a lower limit of precision in estimating individual body masses. In general, it is much more preferable to determine at least broad locomotor affinities, and thus appropriate modern reference groups, before applying body mass estimation equations. However, some structural dimensions are less sensitive to locomotor distinctions than others; for example, proximal tibial articular M-L breadth is apparently "locomotor blind" regarding body mass estimation within the present study sample. In other cases where locomotor affiliation is uncertain, mean estimates from different reference groups can be used, while for some dimensions no estimation should be attempted. The techniques are illustrated by estimating the body masses of four fossil anthropoid specimens of Proconsul nyanzae, Proconsul heseloni, Morotopithecus bishopi, and Theropithecus oswaldi.     

Toxoplasma gondii myosin A and its light chain: a fast, single-headed, plus-end-directed motor

Successful host cell invasion is a prerequisite for survival of the obligate intracellular apicomplexan parasites and establishment of infection. Toxoplasma gondii penetrates host cells by an active process involving its own actomyosin system and which is distinct from induced phagocytosis. Toxoplasma gondii myosin A (TgMyoA) is presumed to achieve power gliding motion and host cell penetration by the capping of apically released adhesins towards the rear of the parasite. We report here an extensive biochemical characterization of the functional TgMyoA motor complex. TgMyoA is anchored at the plasma membrane and binds a novel type of myosin light chain (TgMLC1). Despite some unusual features, the kinetic and mechanical properties of TgMyoA are unexpectedly similar to those of fast skeletal muscle myosins. Microneedle-laser trap and sliding velocity assays established that TgMyoA moves in unitary steps of 5.3 nm with a velocity of 5.2 microm/s towards the plus end of actin filaments. TgMyoA is the first fast, single-headed myosin and fulfils all the requirements for power parasite gliding.