Evolutionary dynamics of the kinetochore network in eukaryotes as revealed by comparative genomics

During eukaryotic cell division, the sister chromatids of duplicated chromosomes are pulled apart by microtubules, which connect via kinetochores. The kinetochore is a multiprotein structure that links centromeres to microtubules, and that emits molecular signals in order to safeguard the equal distribution of duplicated chromosomes over daughter cells. Although microtubule‐mediated chromosome segregation is evolutionary conserved, kinetochore compositions seem to have diverged. To systematically inventory kinetochore diversity and to reconstruct its evolution, we determined orthologs of 70 kinetochore proteins in 90 phylogenetically diverse eukaryotes. The resulting ortholog sets imply that the last eukaryotic common ancestor (LECA) possessed a complex kinetochore and highlight that current‐day kinetochores differ substantially. These kinetochores diverged through gene loss, duplication, and, less frequently, invention and displacement. Various kinetochore components co‐evolved with one another, albeit in different manners. These co‐evolutionary patterns improve our understanding of kinetochore function and evolution, which we illustrated with the RZZ complex, TRIP13, the MCC, and some nuclear pore proteins. The extensive diversity of kinetochore compositions in eukaryotes poses numerous questions regarding evolutionary flexibility of essential cellular functions.     

Estimating Fundamental Host Range: a Host-Specificity Study of a Potential Biocontrol Agent for Prosopis Species (Leguminosae)

One approach to predicting non-target attack by potential biological control agents is to first describe their fundamental host ranges and then to predict how it will be expressed under postrelease field conditions. In this paper, we illustrate how the fundamental host range can be estimated experimentally by excluding possible limiting factors such as time-dependent effects. The example we use is a host-specificity study of a leaf-tying moth (Gelechiidae: Evippe sp. #1) which was being assessed for the biological control of mesquite (Leguminosae: Prosopis spp.) in Australia. Females oviposited all eggs on plants, mostly into cracks and fissures. First instar larvae leaf-mined and subsequent instars leaf-tied. Oviposition was not host-specific in cage trials, although ten times more eggs were laid on Prosopis than on non-targets. The fundamental host range for initiation of larval feeding was restricted to Prosopis and Leucaena leucocephala which both belong to the same tribe, and the fundamental host range for complete larval development was restricted to Prosopis . We predict that if released in Australia Evippe sp. #1 will only attack Prosopis spp., although low levels of 'indiscriminate' oviposition might occur on other taxa, and might result in initiation of feeding on L. leucocephala .     

Species-abundance-biomass responses by estuarine macrobenthos to sediment chemical contamination

Macrobenthic community responses can be measured through concerted changes in univariate metrics,including species richness, total abundance, and totalbiomass. The classic model of pollution effects onmarine macrobenthic communities recognizes thatspecies/abundance/biomass (SAB) curves varydistinctively in a nonlinear manner with the magnitudeof organic enrichment. For example, at moderatelevels of organic enrichment, small-bodiedopportunistic species boost the abundance curve, whilespecies richness falls. Ratios among the metrics formuseful indicators of how the community changes withorganic enrichment. However, the classic SAB model isbased on organic enrichment effects over small spatialand temporal scales, and the applicability of the SABmodel to sediment chemical contamination and acrossbroad natural estuarine gradients is largely unknown. Here, SAB responses were examined with respect toprimary gradients in metals and organic chemicalsbased on an extensive dataset comprising 319 estuarinesites from throughout the northern Gulf of Mexico. Each SAB metric was first adjusted with respect to thethree primary natural estuarine gradients, salinity,depth, and sediment silt/clay content. Adjusted SABrelationships varied in their details with respect todifferent classes of sediment contamination, but alltypes of SAB stress responses appear to exhibitsimilar basic characteristics. As in the SAB model,all three SAB metrics were notably low at the highestconcentrations of both metal and organic-chemicalcontaminants. Moreover, rapid decreases in the B/Aratio with increasing contamination supported theconcept that relatively long-lived, large-bodied,equilibrium taxa decline markedly at highconcentrations of toxicants.     

Enantioselective retardation of rac-propranolol from matrices containing cellulose derivatives

The dissolution characteristics of propranolol enantiomers from tablet formulations containing cellulose, or one of eight cellulose derivatives, were determined under a range of conditions. The derivatives examined were: cellulose tris(phenylcarbamate) (1), cellulose tris(2,3-dichlorophenylcarbamate) (2), cellulose tris 2,4-dichlorophenylcarbamate (3), cellulose tris(2,6-dichlorophenylcarbamate) (4), cellulose tris(2,3-dimethylphenylcarbamate) (5), cellulose tris(3,4-dichlorophenylcarbamate) (6), cellulose tris (3,5-dichlorophenylcarbamate) (7), cellulose tris(3,5-dimethylphenylcarbamate) (8). In water at 25°C, the release rates of (-)R-propranolol were generally greater than those of (-)-S-propranolol, although these differences were not always statistically significant; only compounds 5 and 8 demonstrated significant enantioselectivity. Using compound 8 in further experiments, statistically significant stereoselective dissolution of propranolol HCl was observed in buffer pH 7.4 at 25°C (intrinsic dissolution rates: 0.41 ± 0.01 mgcm²min⁻¹ for R-propranolol and 0.30 ± 0.02 mgcm²min⁻¹ for S-propranolol; P = 0.003). The cumulative amounts of enantiomers released at every time point were also found to be statistically significant (mean ratio R:S 1.25 ± 0.05). The observed low stereoselectivity of 8 with propranolol base was probably attributable to low solubility in pH 7.4 buffer, although stereoselective release did increase with time. This suggested that there is a relationship between stereoselectivity and contact time in an aqueous environment. Results also suggested that increased temperature may affect the release process as well as stereoselective interactions of 8 with individual enantiomers. To conclude, differential release of rac-propranolol from cellulose derivative matrices has been demonstrated, which supports the principle of stereoselective retardation as a potential means of stereoselective drug delivery for solid dosage forms. Chirality 9:307–312, 1997. © 1997 Wiley-Liss, Inc.     

Direct resolution of propranolol and bupranolol by thin-layer chromatography using cellulose derivatives as stationary phase

Cellulose triphenylcarbamate derivatives have been used as stationary phases for resolution of the enantiomers of the β-blockers propranolol and bupranolol by TLC. The derivatives examined were: cellulose trisphenylacarbamate (1), cellulose tris(2,3-dichlorophenyl carbamate) (2), cellulose tris(2,4-dichlorophenyl carbamate) (3), cellulose tris(2,6-dichlorophenyl carbamate) (4), cellulose tris (2,3-dimethylphenyl carbamate) (5), cellulose tris(3,4-dichlorophenyl carbamate) (6), cellulose tris(3,5-dichlorophenyl carbamate) (7), and cellulose tris(3,5-dimethylphenyl carbamate) (8). A variety of mobile phases were used to achieve useful separations and the effects of solvent polarity are also discussed. The best resolution of rac-propranolol was obtained on CSP 8 (R_fR = 0.26, R_fS = 0.06, α = 4.33) in mobile phase hexane:propan-2-ol (80:20 v/v). The best resolution of rac-bupranolol was obtained on CSP 5 (R_fR = 0.29, R_fS = 0.09, α = 3.22) in mobile phase hexane:propan-2-ol (80:20 v/v). These results demonstrated the potential of cellulose triphenylcarbamates as chiral stationary phases in TLC and indicate that this is potentially a useful method for the direct, simple, and rapid (within 30 min) resolution of racemates in the analytical control of enantiomeric purity. Physical aspects such as problems in cracking of the CSP, adhesion to plate, and interference of spot detection due to triphenylcarbamate chromphores are also discussed, along with the method employed to overcome them. Chirality 9:139–144, 1997. © 1997 Wiley-Liss, Inc.     

Thin-layer chromatographic separation of intermediates of the pyridine nucleotide cycle

A rapid thin-layer chromatographic procedure for separation of the compounds comprising the intermediates in the salvage pathway known as the pyridine nucleotide cycle plus quinolinic acid and the reduced forms of nicotinamide adenine dinucleotide and nicotinamide adenine dinucleotide phosphate is described. The method utilizes silica gel high-performance thin-layer plates and a mobile phase of methanol, tetrabutylammonium hydroxide, and acetonitrile. The time required for analysis is greatly reduced and results in greater than 96% purity of each migrating compound.     

MINUTES OF THE ANNUAL GENERAL MEETING OF THE SOUTH AFRICAN ORNITHOLOGICAL SOCIETY,HELD AT CAPE TOWN, 26 JUNE 1959

Brown, C. J. 1989. Plumages and measurements of the Bearded Vulture in southern Africa. Ostrich 60: 165–171.

Four different age classes of the southern African Bearded Vulture Gypaetus barbatus are recognized and their plumages described: juvenile (3–24 months old), immature (24–45 months), subadult (45–60 months) and adult (60+ months). There was no significant difference in size between adult male and female birds. Adults were larger than juvenile birds in bill width, beard length, wingspan and mass, and had a higher aspect ratio and wing loading, while juvenile birds were larger than adults in the length of their outer rectrices, tail area, wing breadth and wing area. These features are considered to be adaptive to young birds inexperienced in flying. Immature and subadult birds were intermediate in size between juveniles and adults. Bearded Vultures differ from other large raptors in two sets of physical characteristics, (a) those adapted to cold, mountainous habitat, e.g. feathered head and face, unusually long wings, a high aspect ratio and a particularly long tail, and (b) those adapted to their diet of mainly bones, e.g. wide gape, beard and relatively long talons for carrying food.     

Growth of Natural Yeast Flora during the Fermentation of Inoculated Wines

The growth of yeasts that occur naturally in grape juice was quantitatively examined during the fermentation of four wines that had been inoculated with Saccharomyces cerevisiae. Although S. cerevisiae dominated the wine fermentations, there was significant growth of the natural species Kloeckera apiculata, Candida stellata, Candida colliculosa, Candida pulcherrima, and Hansenula anomala.     

Discontinuous DNA synthesis by purified mammalian proteins

Five proteins purified from mouse cells acting together efficiently convert a single-stranded circular DNA template to covalently closed duplex circle by a discontinuous mechanism. DNA polymerase alpha/primase with the assistance of alpha accessory factor covers the single-stranded circle with RNA-primed DNA fragments. Primers are removed by a combination of RNase H-1 and a 5'-exonuclease that was identified by its ability to complete this in vitro system. The 5'-exonuclease is required to remove residual one or two ribonucleotides at the primer/DNA junction that are resistant to RNase H-1. Gap filling is by the DNA polymerase alpha/primase, and DNA ligase I converts the DNA fragments to continuous strand. The concerted action of the five proteins emulates synthesis of the staging strand at the replication fork.