Implementation of two high through-put techniques in a novel application: detecting point mutations in large EMS mutated plant populations

Background  

The establishment of mutant populations together with the strategies for targeted mutation detection has been applied successfully to a large number of organisms including many species in the plant kingdom. Considerable efforts have been invested into research on tomato as a model for berry-fruit plants. With the progress of the tomato sequencing project, reverse genetics becomes an obvious and achievable goal.     

Leishmania donovani-derived lipophosphoglycan plus BCG induces a Th1 type immune response but does not protect Syrian golden hamsters (Mesocricetus auratus) and BALB/c mice against Leishmania donovani

The efficacy of Leishmania donovani-derived lipophosphoglycan (LPG) plus Mycobacterium bovis Bacille Calmette-Guerin (BCG) as a vaccine candidate against visceral leishmaniosis in susceptible BALB/c mouse and Syrian golden hamster (Mesocricetus auratus) models was investigated. Following a triple vaccination with a total dose of 150 microl BCG plus 60 microg or 30 microg of LPG for hamsters and BALB/c mice respectively, there were no noticeable side effects both locally and systemically; implying that the molecule was safe at this dosage level. Vaccinated animals demonstrated an activation of both the humoral as well as cell-mediated responses to LPG, which correlated with resistance against the disease. Protection by LPG plus BCG, was however, poor as the remaining immunized animals showed disease progression leading to severity of the disease as illustrated by emaciation, mass loss and heavy splenic parasitaemia in hamsters. These data nevertheless suggest that it may be rewarding to further evaluate the potential of LPG as a vaccine candidate in leishmaniosis using other adjuvants, which may enhance its immunogenicity.     

Leishmania major infections in Phlebotomus duboscqi fed on murine models immunized with L. major subcellular antigens and sandfly gut antigens

The ability of antibodies in bloodmeals of mice and hamsters immunized with Leishmania major subcellular fractions and sandfly (Phlebotomus duboscqi) gut antigens to inhibit development of L. major in its vector P. duboscqi was examined. Antibodies from animals immunized with either L. major subcellular fractions alone or sandfly gut antigen alone were not very effective in inhibiting development of L. major in the sandfly. When P. duboscqi were fed on blood from animals immunized with both parasite flagella and sandfly gut antigen, development of L. major was significantly inhibited (P<0,05). Control sandflies fed on naive animals displayed a normal pattern of parasite development to the metacyclic stage. Electron microscopy studies showed that one of the mechanisms through which antisandfly gut antibody can cause inhibition of parasite development is by lysing sandfly gut epithelium. This study has demonstrated that it is possible to reduce transmission of leishmaniosis through immunization against both the parasite and its sandfly vector.     

Isolation and pathogenicity of Australian strains of Eimeria praecox and Eimeria mitis

Objective To determine the presence of E praecox and E mitis in Australia, to isolate representative strains of these species from chickens and determine their pathogenicity.
Design Morphological, physiological and cross protection studies were undertaken to confirm the identity of Australian isolates of E praecox and E mitis.
Procedure Oocysts were isolated from a backyard flock at Jimboomba, southeastern Queensland and numbers of E praecox and E mitis enriched by passage in chickens immune to five other species of poultry Eimeria . Oocysts of mean conformation and size of the two species were purified by single oocyst passage. Two isolates that closely matched recorded parameters for E praecox and E mitis were selected and designated JP and JM respectively. The cross protection between the isolates and E acervulina was determined by infection and challenge experiments. The virulence of the two isolates was determined by comparing weight gains of groups of birds inoculated with JP isolate or JM isolate with untreated groups.
Results Isolates JP and JM most closely matched recorded parameters of E praecox and E mitis respectively. Groups of chickens, previously infected with JP and JM isolates, showed no significant protection against infection with E acervulina . In a separate trial, groups of susceptible chickens inoculated with 10⁵ oocysts of JP and JM isolates showed significantly reduced weight gains compared with untreated controls.
Conclusion Isolates JP and JM are E praecox and E mitis respectively, confirming the presence of these species in Australia. These isolates were found capable of causing significant reductions in weight gains in susceptible chickens.     

Supra-canonical 26Al/27Al and the residence time of CAIs in the solar protoplanetary disk

The canonical initial 26Al/27Al ratio of 4.5 x 10(-5) has been a fiducial marker for the beginning of the solar system. Laser ablation and whole-rock multiple-collector inductively coupled plasma-source mass spectrometry magnesium isotope analyses of calcium- and aluminum-rich inclusions (CAIs) from CV3 meteorites demonstrate that some CAIs had initial 26Al/27Al values at least 25% greater than canonical and that the canonical initial 26Al/27Al cannot mark the beginning of solar system formation. Using rates of Mg diffusion in minerals, we find that the canonical initial 26Al/27Al is instead the culmination of thousands of brief high-temperature events incurred by CAIs during a 10(5)-year residence time in the solar protoplanetary disk.