135 research outputs found
An improved single-step lysis protocol to measure luciferase bioluminescence in Plasmodium falciparum
This report describes the optimization and evaluation of a simple single-step lysis protocol to measure luciferase bioluminescence from genetically modified Plasmodium falciparum. This protocol utilizes a modified commercial buffer to improve speed of assay and consistency in the bioluminescence signal measured by reducing the manipulation steps required to release the cytoplasmic fraction. The utility of this improved assay protocol is demonstrated in typical assays that explore absolute and temporal gene expression activity
Comparison of the absolute and relative efficiencies of electroporation-based transfection protocols for Plasmodium falciparum.
BACKGROUND: Several electroporation protocols exist to transfect exogenous DNA into Plasmodium falciparum. To date, however, only a subjective analysis of their relative efficiencies has been reported. METHODS: A time-course of luciferase reporter expression is used to provide an objective quantitative analysis of the absolute efficiency of three electroporation techniques; direct electroporation of ring stage infected erythrocytes, preloading of erythrocytes and a novel "double-tap" protocol that combines both approaches. RESULTS: Preloading of erythrocytes shows a mean efficiency of 9.59 x 10(-6), some 5-180 fold more efficient than matched experiments utilizing the "double-tap" and direct electroporation of ring stage infected erythrocytes alone, respectively. CONCLUSION: Evidence presented here provides the first quantitative assessment of both the absolute and relative efficiencies of a key molecular tool used to study the biology and pathogenesis of this important human pathogen
Testing Superstring Theories with Gravitational Waves
We provide a simple transfer function that determines the effect of an early
matter dominated era on the gravitational wave background and show that a large
class of compactifications of superstring theory might be tested by
observations of the gravitational wave background from inflation. For large
enough reheating temperatures > 10^9 \GeV the test applies to all models
containing at least one scalar with mass < 10^{12}\GeV that acquires a large
initial oscillation amplitude after inflation and has only gravitational
interaction strength, i.e., a field with the typical properties of a modulus.Comment: 5 pages 2 figures, v2: changes in presentation, refs revised, matches
version in print in PR
Evaluation of bioluminescence-based assays of anti-malarial drug activity
Transgenic Plasmodium falciparum expressing luciferase offers an attractive bioluminescence-based assay platform for the investigation of the pharmacological properties of anti-malarial drugs. Here a side-by-side comparison of bioluminescence and fluorescence-based assays, utilizing a luciferase reporter cassette that confers a strong temporal pattern of luciferase expression during the S-phase of intraerythrocytic development, is reported
Gravitino Dark Matter in Tree Level Gauge Mediation with and without R-parity
We investigate the cosmological aspects of Tree Level Gauge Mediation, a
recently proposed mechanism in which the breaking of supersymmetry is
communicated to the soft scalar masses by extra gauge interactions at the tree
level. Embedding the mechanism in a Grand Unified Theory and requiring the
observability of sfermion masses at the Large Hadron Collider, it follows that
the Lightest Supersymmetric Particle is a gravitino with a mass of the order of
10 GeV. The analysis in the presence of R-parity shows that a typical Tree
Level Gauge Mediation spectrum leads to an overabundance of the Dark Matter
relic density and a tension with the constraints from Big Bang Nucleosynthesis.
This suggests to relax the exact conservation of the R-parity. The underlying
SO(10) Grand Unified Theory together with the bounds from proton decay provide
a rationale for considering only bilinear R-parity violating operators. We
finally analyze the cosmological implications of this setup by identifying the
phenomenologically viable regions of the parameter space.Comment: 28 pages, 5 figures. References added. To appear in JHE
Dark Radiation and Dark Matter in Large Volume Compactifications
We argue that dark radiation is naturally generated from the decay of the
overall volume modulus in the LARGE volume scenario. We consider both
sequestered and non-sequestered cases, and find that the axionic superpartner
of the modulus is produced by the modulus decay and it can account for the dark
radiation suggested by observations, while the modulus decay through the
Giudice-Masiero term gives the dominant contribution to the total decay rate.
In the sequestered case, the lightest supersymmetric particles produced by the
modulus decay can naturally account for the observed dark matter density. In
the non-sequestered case, on the other hand, the supersymmetric particles are
not produced by the modulus decay, since the soft masses are of order the heavy
gravitino mass. The QCD axion will then be a plausible dark matter candidate.Comment: 27 pages, 4 figures; version 3: version published in JHE
Rituximab retherapy in patients with relapsed aggressive B cell and mantle cell lymphoma
Neither effective salvage regimens nor the outcome and response to retherapy with rituximab containing chemotherapy have been defined for rituximab pre-treated patients with relapsing aggressive lymphoma. We report here a single-centre retrospective outcome analysis of second-line immunochemotherapy with rituximab. In 28 patients with relapsed or refractory diffuse large B cell lymphomas, first-line immunochemotherapy had induced objective responses in 18 patients. Nine of 28 patients responded to rituximab containing salvage therapy, leading to a median overall survival of 243Â days after start of second immunochemotherapy. Long-term disease free survivors (1,260 and 949Â days) were restricted to the group of twelve patients that had received allogeneic stem cell transplantation as consolidation therapy. In 21 patients with relapsed mantle cell lymphomas (MCL), 19 patients had reached remissions with first-line therapy. Of those, 16 patients experienced responses to salvage therapy with a median overall survival of 226Â days. Noteworthy, none of patients with initial non-responding disease reached a remission with second immunochemotherapy. Seven patients with MCL stayed free from progression after high-dose therapy with autologous or allogeneic stem cell transplantation in two and five cases, respectively. In summary, responses to repeated immunotherapy with rituximab were observed in approximately one third and two thirds of initially responding patients with aggressive B cell lymphoma and mantle cell lymphoma, respectively, but not in primarily refractory disease. Lasting remissions were achieved only by high-dose chemotherapy with stem cell transplantation
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