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Biogenesis of Yersinia pestis PsaA in recombinant attenuated Salmonella Typhimurium vaccine (RASV) strain

FEMS MICROBIOLOGY LETTERS, Issue 2 2010
Ascención Torres-Escobar
Abstract Yersinia pestis PsaA is an adhesin important for the establishment of bacterial infection. PsaA synthesis requires the products of the psaEFABC genes. Here, by prediction analysis, we identified a PsaA signal sequence with two signal peptidase (SPase) cleavage sites, type-I and type-II (SPase-I and SPase-II). By Edman degradation and site-directed mutagenesis, the precise site for one of these Spase-I PsaA cleavage sites was located between alanine and serine at positions 31 and 32, respectively. Yersinia pestis psaA expression and the role of the PsaB and PsaC proteins were evaluated in recombinant attenuated Salmonella Typhimurium vaccine strains. PsaA was detected in total extracts as a major 15-kDa (mature) and 18-kDa (unprocessed) protein bands. PsaA synthesis was not altered by a ,A31,,S32 double-deletion mutation. In contrast, the synthesis of PsaA (,A31,,S32) in Y. pestis and delivery to the supernatant was decreased. Otherwise, substitution of the amino acid cysteine at position 26 by valine involved in the SPase-II cleavage site did not show any effect on the secretion of PsaA in Salmonella and Yersinia. These results help clarify the secretion pathway of PsaA for the possible development of vaccines against Y. pestis. [source]


Restricted fish feeding reduces cod otolith opacity

JOURNAL OF APPLIED ICHTHYOLOGY, Issue 2 2008
H. Hřie
Summary The purpose of this work was to examine the effect of reduced feeding and constant temperature on cod otolith opacity. Three groups of juvenile cod were given restricted food rations at different times for 4 months, resulting in depressed somatic growth. Otolith opacity was measured on pictures of the otolith sections. The otolith carbonate deposited during the experimental period was generally opaque compared to the more translucent otolith material deposited prior to and after the experimental period, when the fish were kept in a pond and in sea-cages at higher temperatures. Large variations in otolith opacity were found between individual fish both within groups and between groups. In two of the three groups significantly more translucent otolith material was deposited in response to reduced feeding. Our results show that variations in feeding and hence fish growth resulted in variation in otolith opacity, but the effect was minor compared to that of variations in ambient temperature. The combined influence of these effects, which both act on fish metabolism, are most likely controlling the seasonal opacity changes observed in wild fish. Our results help explain the variations seen in fish at constant temperatures. [source]


The spread of apomixis and its effect on resident genetic variation

JOURNAL OF EVOLUTIONARY BIOLOGY, Issue 5 2007
S. ADOLFSSON
Abstract In a simulation model we investigated how much of the initial genetic variation that is retained in a population after a dominant mutation has brought apomixis to fixation in it. A marker allele associated with the apomixis mutation is generally retained after the fixation of apomixis, particularly if the two alleles are closely linked. The spread of asexuality, however, normally leads to almost no loss of genetic variation, neither with respect to cytotypes nor with respect to genotypes. This holds for large populations and apomixis mutants with strong pollen production. In smaller populations, and with apomicts with reduced pollen production, the outcome is more variable, ranging from no genetic variation retained to only weakly reduced variability compared with the initial state. These results help explain the high genetic variability in many apomicts. They also imply that natural selection will have many genotypes to act on even after the spread of apomixis. [source]


High-pressure granulites in the Sanggan area, North China craton: metamorphic evolution, P,T paths and geotectonic significance

JOURNAL OF METAMORPHIC GEOLOGY, Issue 8 2002
J. H. Guo
Abstract High-pressure basic granulites are widely distributed as enclaves and sheet-like blocks in the Huaian TTG gneiss terrane in the Sanggan area of the Central Zone of the North China craton. Four stages of the metamorphic history have been recognised in mineral assemblages based on inclusion, exsolution and reaction textures integrated with garnet zonation patterns as revealed by compositional maps and compositional profiles. The P,T conditions for each metamorphic stage were obtained using thermodynamically and experimentally calibrated geothermobarometers. The low-Ca core of growth-zoned garnet, along with inclusion minerals, defines a prograde assemblage (M1) of garnet + clinopyroxene + plagioclase + quartz, yielding 700 °C and 10 kbar. The peak of metamorphism at about 750,870 °C and 11,14.5 kbar (M2) is defined by high-Ca domains in garnet interiors and inclusion minerals of clinopyroxene, plagioclase and quartz. Kelyphites or coronas of orthopyroxene + plagioclase ± magnetite around garnet porphyroblasts indicate garnet breakdown reactions (M3) at conditions around 770,830 °C and 8.5,10.5 kbar. Garnet exsolution lamellae in clinopyroxene and kelyphites of amphibole + plagioclase around garnet formed during the cooling process at about 500,650 °C and 5.5,8 kbar (M4). These results help define a sequential P,T path containing prograde, near-isothermal decompression (ITD) and near-isobaric cooling (IBC) stages. The clockwise hybrid ITD and IBC P,T paths of the HP granulites in the Sanggan area imply a model of thickening followed by extension in a collisional environment. Furthermore, the relatively high-pressures (6,14.5 kbar) of the four metamorphic stages and the geometry of the P,T paths suggest that the HP granulites, together with their host Huaian TTG gneisses, represent the lower plate in a crust thickened during collision. The corresponding upper-plate might be the tectonically overlying Khondalite series, which was subjected to medium- to low-pressure (MP/LP: 7,4 kbar) granulite facies metamorphism with a clockwise P,T path including an ITD segment. Both the HP and the MP/LP granulite facies events occurred contemporaneously at c. 1.90,1.85 Ga in a collisional environment created by the assembly process of the North China craton. [source]


Cell cycle regulator Cdc14 is expressed during sporulation but not hyphal growth in the fungus-like oomycete Phytophthora infestans

MOLECULAR MICROBIOLOGY, Issue 2 2003
Audrey M. V. Ah Fong
Summary Cdc14 proteins are important regulators of mitosis and the cell cycle. These phosphatases have been studied previously only in yeasts and metazoans, which grow by fission or budding. Here we describe a homologue (piCdc14) from the oomycete Phytophthora infestans, a primitive eukaryote lacking a classical cell cycle. PiCdc14 complements a cdc14ts mutant of Saccharomyces cerevisiae and may function like other Cdc14 proteins, but displays a strikingly different pattern of expression. Whereas previously studied Cdc14 genes are constitutively transcribed, piCdc14 is not expressed during normal growth but instead only during asexual sporulation. In transformants of P. infestans expressing a fusion between the piCdc14 promoter and the ,-glucuronidase reporter, expression was first detected in sporangiophore initials, persisted in sporangiophores bearing immature sporangia, and later became restricted to mature sporangia. After germination, expression ended a few hours before the resumption of mitosis in hyphae emerged from the spores. Homology-dependent silencing experiments supported an essential role of piCdc14 in sporulation. It is proposed that the function of piCdc14 may be to synchronise nuclear behaviour during sporulation and maintain dormancy in spores until germination. These results help illuminate the process of sporulation in oomycetes and the evolution of the cell cycle in eukaryotes. [source]


Variation in 16S-23S rRNA intergenic spacer regions among Bacillus subtilis 168 isolates

MOLECULAR MICROBIOLOGY, Issue 1 2001
Yevette J. Shaver
The genome of the Bacillus subtilis 168-type strain contains 10 ribosomal RNA (rRNA) operons. In the intergenic spacer region (ISR) between the 16S and 23S rRNA genes, five rRNA operons, rrnI-H-G and rrnJ-W, lack a trinucleotide signature region. Precise determination of molecular weight (MW), using electrospray mass spectrometry (MS), of the polymerase chain reaction (PCR) products from a segment of the ISR from the 168-type strain and B. subtilis 168-like strain 23071 demonstrated 114 and 111 basepair (bp) PCR products (due to the presence or absence of the insert in the operons) as predicted from sequence. However, PCR of the ISR segment for five other B. subtilis 168 isolates generated only a 114 bp PCR product, suggesting the presence of the trinucleotide signature region in all rRNA operons for these strains. Additional genetic variability between the seven B. subtilis 168 isolates was demonstrated by restriction fragment length polymorphism (RFLP) of the rRNA operons, with three distinct patterns found upon Southern blot analysis. The 168-type strain and three others (23066, 23067, and 23071) exhibited the same Southern pattern. Thus, operon deletion is not responsible for the absence of a 111 bp product on MS analysis for strains 23066 and 23067. Restriction analysis confirmed the presence of the trinucleotide signature region in the ISR of all rRNA operons for five B. subtilis 168 isolates; sequencing of rrnW/H from a representative strain also upheld this finding. These results help provide a better understanding of variations in sequence, operon number and chromosomal organization, both within a genome and among isolates of B. subtilis subgroup 168. It is also hypothesized that the presence of the trinucleotide insert in certain rRNA operons may play a role in rRNA maturation and protein synthesis. [source]


RESEARCH PRODUCTIVITY OF AUSTRALIAN ACADEMIC ECONOMISTS: HUMAN-CAPITAL AND FIXED EFFECTS,

AUSTRALIAN ECONOMIC PAPERS, Issue 1 2007
JOAN R. RODGERS
This study investigates why some economics departments in Australian universities are more research productive than others. The hypothesis is simple: research productivity depends upon the human capital of department members and the department-specific conditions under which they work. A Tobit model is used to estimate the magnitude of the two effects. Both are found to be important. Our results help explain why a small number of departments consistently outperform the others in studies that rank Australian economics departments according to research output. [source]