Cell mediated immune response elicited in mice after immunization with the P64k meningococcal protein: epitope mapping

The P64k protein of Neisseria meningitidis has been reported as an immunological carrier for weak immunogens. This investigation was aimed at characterizing the T-cell response produced in primed mice and at identifying T helper cell epitopes within this molecule. BALB/c mice subcutaneously immunized with the recombinant antigen provided inguinal lymph node cells (LNC) that proliferated in the presence of P64k in a dose-dependent manner. Proliferating cells secreted IL-4 while the concentration of IL-12 remained unaltered in the culture supernatant. By testing a panel of 59 overlapping synthetic peptides spanning the entire sequence of the antigen a T-cell determinant was localized. Prime-boost and lymphoproliferation experiments, conducted with highly purified synthetic peptides, confirmed that the segment including amino acids 470-485 comprises a T-cell epitope within the P64k molecule.     

Coralline algae indicate Pleistocene evolution from deep,open platform to outer barrier reef environments in the northern Great Barrier Reef margin

Coralline algae from a drill core in Ribbon Reef 5 have been used to interpret changes in the depositional palaeoenvironments in the northern Australian Great Barrier Reef over the last 790 ka. Three main coralline algal assemblages, each dominated by members of a particular subfamily, have been distinguished by quantitative analysis: (1) mastophoroid assemblages, usually occurring as crusts on corals, are typical of the shallowest reef settings; (2) lithophylloid assemblages within algal nodules may represent shallow-water, cooler environments or deeper reef subenvironments; and (3) melobesioid assemblages are characteristic of deeper-water platform areas. The algal assemblages between 96 and 210 m b.s.f. (metre below sea floor) record a fluctuating but progressive shallowing-upwards from deep, outer-platform to shallower, non-reefal depositional environments. Two intervening episodes dominated by mastophoroids represent two phases of reef growth. The section above 96 m b.s.f. comprises several stacked reefs in which mastophoroid crusts similar to the present-day shallow-water assemblages predominate.Electronic Supplementary Material Supplementary material is available for this article at .Communicated by: Geological Editor P.K. Swart     

Oxidative stress and antioxidant defenses: a target for the treatment of diseases caused by parasitic protozoa

Parasitic protozoa cause several diseases, affecting hundreds of millions, particularly in underdeveloped countries. Although these organisms are eukaryotic cells, some of them present major differences with their mammalian host in selected metabolic pathways. These differences may be exploited as targets for developing better pharmacological agents for the treatment of specific parasitic diseases. This review describes some of the differences in terms of antioxidant defenses between these organisms and their mammalian host, which may provide useful targets for the treatment of these diseases. Some of the potential targets are: (i). iron metabolism in Plasmodium, (ii). the presence of a Fe-containing form of superoxide dismutase in trypanosomatids and malaria-causing parasites, (iii). the unique trypanothione-dependent antioxidant metabolism in trypanosomatids, (iv). the ascorbate peroxidase found in Trypanosoma cruzi and perhaps present in other trypanosomatids.     

Spectrophotometric method for determining gibberellic acid in fermentation broths

A novel method for the quantitative determination of gibberellic acid in fermentation broths has been developed. It is based on the kinetic of the reaction of conversion of gibberellic acid to gibberellenic acid. The method is simple, reliable, faster than most of methods known, and free of the interferences which commonly affect spectrophotometric methods currently in use. Its threshold sensitivity is 0.1 g and its accuracy is greater than 97% for concentrations of gibberellic acid ranging from 0.1 to 1 g l(-1).     

Active and inactive ecto-5'-nucleotidase variants in liver of control and dystrophic Lama2dy mice

The ecto-5'-nucleotidase (eNT) activity and the eNT protein content in liver of normal and merosin-deficient dystrophic Lama2dy mice were studied. After the solubilization procedure, the eNT activity in the final extract was 9.2+/-2.5U/mg (nmol of phosphate released from AMP per min and per mg protein) in normal liver, and it rose to 16.1+/-3.9U/mg (P=0.005) in dystrophic liver. The increase of activity was less pronounced in Lama2dy liver (1.7-fold) than the one reported in muscle (four-fold), which probably reflects the lower content of merosin in liver. Similarly to muscle, liver contained active and inactive eNT, as demonstrated by the higher level of immunoreactive protein in normal than in dystrophic liver in Western blots performed with samples containing the same units of eNT activity. PNGase F digestion decreased the size of liver and muscle eNT from 72 and 69kDa, to 63 and 60kDa. Oligoglycan cleavage did not alter eNT activity or the sedimentation coefficient, revealing that oligosaccharides are not required for catalysis or for maintaining the dimeric structure. The eNT protein content in samples of normal liver decreased by 55 or 80% after the trypsinolysis of native or deglycosylated enzyme, but the activity did not change. Such a high proportion of inactive eNT is unlikely to come from aged enzyme, which suggests the involvement of inactive enzyme in non-catalytic actions.     

Effects of Nutrient Limitation on Aboveground Carbon Dynamics during Tropical Dry Forest Regeneration in Yucatán,Mexico

Tree growth (as diameter increment), litterfall production, and litter biomass were studied in two secondary tropical dry forests of the Yucatán Peninsula under four treatments of nutrient addition. The studys objective was to assess how variations in the nutrient supply affect aboveground net primary production and carbon (C) accumulation on the floor of two forests in different stages of regeneration. The study included an area of young forest (10 years old) with phosphorus (P)-poor soils and an area of old forest (around 60 years old) where soil P was comparatively less limiting. Four replicate plots (12 × 12 m) at each forest were either left intact (controls) or fertilized with nitrogen (N), P, or N plus P during 3 consecutive years. After 3 years of fertilization, relaxation of the constraints on nutrient limitation resulted in increased trunk growth rates at both the young and old forests. This effect was more pronounced with the addition of P or N plus P (trunk growth doubled with respect to controls), whereas N addition increased tree growth by 60% in comparison to trees in plots without nutrient supplements. In both forests, there were no significant differences in litterfall production among treatments during the first 2 years after fertilization. In the 3rd year of nutrient addition, litterfall production was significantly higher in plots fertilized with N plus P compared to control plots at both forest sites; however, changes in litterfall were not accompanied by litter accumulation in the floor of the two forests. The results of this study support the hypothesis that there is nutrient limitation during tropical dry forest regeneration. They further show that it may be maintained in the long term during secondary succession.     

DNA double-strand breaks, recombination and synapsis: the timing of meiosis differs in grasshoppers and flies

The temporal and functional relationships between DNA events of meiotic recombination and synaptonemal complex formation are a matter of discussion within the meiotic field. To analyse this subject in grasshoppers, organisms that have been considered as models for meiotic studies for many years, we have studied the localization of phosphorylated histone H2AX (gamma-H2AX), which marks the sites of double-strand breaks (DSBs), in combination with localization of cohesin SMC3 and recombinase Rad51. We show that the loss of gamma-H2AX staining is spatially and temporally linked to synapsis, and that in grasshoppers the initiation of recombination, produced as a consequence of DSB formation, precedes synapsis. This result supports the idea that grasshoppers display a pairing pathway that is not present in other insects such as Drosophila melanogaster, but is similar to those reported in yeast, mouse and Arabidopsis. In addition, we have observed the presence of gamma-H2AX in the X chromosome from zygotene to late pachytene, indicating that the function of H2AX phosphorylation during grasshopper spermatogenesis is not restricted to the formation of gamma-H2AX foci at DNA DSBs.     

Genetic demography of Antioquia (Colombia) and the Central Valley of Costa Rica

We report a comparative genetic characterization of two population isolates with parallel demographic histories: the Central Valley of Costa Rica (CVCR) and Antioquia (in northwest Colombia). The analysis of mtDNA, Y-chromosome and autosomal polymorphisms shows that Antioquia and the CVCR are genetically very similar, indicating that closely related parental populations founded these two isolates. In both populations, the male ancestry is predominantly European, whereas the female ancestry is mostly Amerind. In agreement with their isolation, the Amerindian mtDNA diversity of Antioquia and the CVCR is typical of ethnically-defined native populations and is markedly lower than in other Latin American populations. A comparison of linkage disequilibrium (LD) at 18 marker pairs in Antioquia and the CVCR shows that markers in LD in both populations are located at short genetic distances (相似文献   

Effects of GNA transgenic sugarcane on life history parameters of Parallorhogas pyralophagus (Marsh) (Hymenoptera: Braconidae), a parasitoid of Mexican rice borer

Parallorhogas pyralophagus (Marsh) is the principal parasitoid of the Mexican rice borer [Eoreuma loftini (Dyar)], the primary pest of sugarcane in south Texas. Insect resistant transgenic sugarcane producing Galanthus nivalis agglutinin (GNA) was developed to improve control of E. loftini. The present laboratory study addressed whether over two consecutive generations transgenic sugarcane delivered via artificial diet has adverse host-mediated effects on P. pyralophagus life history parameters. The results of this study showed that a number of life history parameters were affected by transgenic sugarcane, whereas others were not affected, and that effects varied between generations. In the first generation, adult longevity was increased by approximately 2 d, and cocoon to adult and egg to adult developmental times were prolonged by approximately 1 d in parasitoids exposed to transgenic sugarcane, whereas effects were not evident on adult size; egg load; egg to cocoon developmental time; rates of gain of longevity and egg load with adult size; and egg, larval, and pupal mortality. However, in the second generation, adult longevity was reduced by approximately 3 d, adult size by approximately 5%, egg load by approximately 24%, and rate of gain of longevity with adult size by approximately 21%, whereas effects were not evident on the rate of gain of egg load with adult size. It was concluded that although GNA transgenic sugarcane, ingested via E. loftini tissues, was not acutely toxic to P. pyralophagus, the sublethal effects on life history parameters measured in this study must be considered in a broader context to determine their possible ecological significance.