In vitro culture of Plasmodium berghei-ANKA maintains infectivity of mouse erythrocytes inducing cerebral malaria

<p>Abstract</p> <p>Background</p> <p>Infection with <it>Plasmodium berghei </it>is a widely used model of murine malaria and a powerful tool for reverse genetic and pathogenesis studies. However, the efficacy of <it>in vitro </it>reinvasion of erythrocytes is generally low, limiting <it>in vitro </it>studies.</p> <p>Methods</p> <p><it>Plasmodium berghei </it>ANKA-infected blood obtained from a susceptible infected mouse was cultured in various conditions and <it>in vitro </it>parasitaemia was measured every day to evaluate the rate of reinvasion.</p> <p>Results</p> <p>High quality culture media were used and reinvasion rates were improved by vigorous orbital shaking of the flask and increasing density of the medium with gelatin.</p> <p>Discussion</p> <p>Using these settings, reinvasion of normal mouse erythrocytes by the parasite was obtained <it>in vitro </it>over two weeks with preservation of the infectivity <it>in vivo</it>.</p

Antimalarial Exposure Delays Plasmodium falciparum Intra-Erythrocytic Cycle and Drives Drug Transporter Genes Expression

BACKGROUND: Multi-drug resistant Plasmodium falciparum is a major obstacle to malaria control and is emerging as a complex phenomenon. Mechanisms of drug evasion based on the intracellular extrusion of the drug and/or modification of target proteins have been described. However, cellular mechanisms related with metabolic activity have also been seen in eukaryotic systems, e.g. cancer cells. Recent observations suggest that such mechanism may occur in P. falciparum. METHODOLOGY/PRINCIPAL FINDINGS: We therefore investigated the effect of mefloquine exposure on the cell cycle of three P. falciparum clones (3D7, FCB, W2) with different drug susceptibilities, while investigating in parallel the expression of four genes coding for confirmed and putative drug transporters (pfcrt, pfmdr1, pfmrp1 and pfmrp2). Mefloquine induced a previously not described dose and clone dependent delay in the intra-erythrocytic cycle of the parasite. Drug impact on cell cycle progression and gene expression was then merged using a non-linear regression model to determine specific drug driven expression. This revealed a mild, but significant, mefloquine driven gene induction up to 1.5 fold. CONCLUSIONS/SIGNIFICANCE: Both cell cycle delay and induced gene expression represent potentially important mechanisms for parasites to escape the effect of the antimalarial drug

Genome-Wide Compensatory Changes Accompany Drug- Selected Mutations in the Plasmodium falciparum crt Gene

Mutations in PfCRT (Plasmodium falciparum chloroquine-resistant transporter), particularly the substitution at amino acid position 76, confer chloroquine (CQ) resistance in P. falciparum. Point mutations in the homolog of the mammalian multidrug resistance gene (pfmdr1) can also modulate the levels of CQ response. Moreover, parasites with the same pfcrt and pfmdr1 alleles exhibit a wide range of drug sensitivity, suggesting that additional genes contribute to levels of CQ resistance (CQR). Reemergence of CQ sensitive parasites after cessation of CQ use indicates that changes in PfCRT are deleterious to the parasite. Some CQR parasites, however, persist in the field and grow well in culture, which may reflect adaptive changes in the parasite genome to compensate for the mutations in PfCRT. Using three isogenic clones that have different drug resistance profiles corresponding to unique mutations in the pfcrt gene (106/1K76, 106/176I, and 106/76I-352K), we investigated changes in gene expression in these parasites grown with and without CQ. We also conducted hybridizations of genomic DNA to identify copy number (CN) changes in parasite genes. RNA transcript levels from 45 genes were significantly altered in one or both mutants relative to the parent line, 106/1K76. Most of the up-regulated genes are involved in invasion, cell growth and development, signal transduction, and transport activities. Of particular interest are genes encoding proteins involved in transport and/or regulation of cytoplasmic or compartmental pH such as the V-type H+ pumping pyrophosphatase 2 (PfVP2), Ca2+/H+ antiporter VCX1, a putative drug transporter and CN changes in pfmdr1. These changes may represent adaptations to altered functionality of PfCRT, a predicted member of drug/metabolite transporter superfamily found on the parasite food vacuole (FV) membrane. Further investigation of these genes may shed light on how the parasite compensates for functional changes accompanying drug resistance mutations in a gene coding for a membrane/drug transporter

WEAVE-StePS: A stellar population survey using WEAVE at WHT

Context. The upcoming new generation of optical spectrographs on four-meter-class telescopes will provide valuable opportunities for forthcoming galaxy surveys through their huge multiplexing capabilities, excellent spectral resolution, and unprecedented wavelength coverage. Aims. WEAVE is a new wide-field spectroscopic facility mounted on the 4.2 m William Herschel Telescope in La Palma. WEAVE-StePS is one of the five extragalactic surveys that will use WEAVE during its first five years of operations. It will observe galaxies using WEAVE MOS (∼950 fibres distributed across a field of view of ∼3 square degrees on the sky) in low-resolution mode (R ∼ 5000, spanning the wavelength range 3660-9590 Å). Methods. WEAVE-StePS will obtain high-quality spectra (S/N ∼ 10 Å -1 at R ∼ 5000) for a magnitude-limited (IAB = 20.5) sample of ∼25 000 galaxies, the majority selected at z ≥ 0.3. The survey goal is to provide precise spectral measurements in the crucial interval that bridges the gap between LEGA-C and SDSS data. The wide area coverage of ∼25 square degrees will enable us to observe galaxies in a variety of environments. The ancillary data available in each of the observed fields (including X-ray coverage, multi-narrow-band photometry and spectroscopic redshift information) will provide an environmental characterisation for each observed galaxy. Results. This paper presents the science case of WEAVE-StePS, the fields to be observed, the parent catalogues used to define the target sample, and the observing strategy that was chosen after a forecast of the expected performance of the instrument for our typical targets. Conclusions. WEAVE-StePS will go back further in cosmic time than SDSS, extending its reach to encompass more than ∼6 Gyr. This is nearly half of the age of the Universe. The spectral and redshift range covered by WEAVE-StePS will open a new observational window by continuously tracing the evolutionary path of galaxies in the largely unexplored intermediate-redshift range

Optoacoustic imaging of the skin

Optoacoustic (OA, photoacoustic) imaging capitalizes on the synergistic combination of light excitation and ultrasound detection to empower biological and clinical investigations with rich optical contrast while effectively bridging the gap between micro and macroscopic imaging realms. State-of-the-art OA embodiments consistently provide images at micron-scale resolution through superficial tissue layers by means of focused illumination that can be smoothly exchanged for acoustic-resolution images at diffuse light depths of several millimetres to centimetres via ultrasound beamforming or tomographic reconstruction. Taken together, this unique multi-scale imaging capacity opens unprecedented capabilities for high-resolution in vivo interrogations of the skin at scalable depths. Moreover, diverse anatomical and functional information is retrieved via dynamic mapping of endogenous chromophores such as haemoglobin, melanin, lipids, collagen, water and others. This, along with the use of non-ionizing radiation, facilitates a clinical translation of the OA modalities. We review recent progress in OA imaging of the skin in preclinical and clinical studies exploiting the rich contrast provided by endogenous substances in tissues. The imaging capabilities of existing approaches are discussed in the context of initial translational studies on skin cancer, inflammatory skin diseases, wounds and other conditions