Efficient generation of mNeonGreen <i>Plasmodium falciparum</i> reporter lines enables quantitative fitness analysis

CRISPR editing has enabled the rapid creation of fluorescent Plasmodium transgenic lines, facilitating a deeper understanding of parasite biology. The impact of genetic perturbations such as gene disruption or the introduction of drug resistance alleles on parasite fitness is typically quantified in competitive growth assays between the query line and a wild type reference. Although fluorescent reporter lines offer a facile and frequently used method to measure relative growth, this approach is limited by the strain background of the existing reporter, which may not match the growth characteristics of the query strains, particularly if these are slower-growing field isolates. Here, we demonstrate an efficient CRISPR-based approach to generate fluorescently labelled parasite lines using mNeonGreen derived from the LanYFP protein in Branchiostoma lanceolatum, which is one of the brightest monomeric green fluorescent proteins identified. Using a positive-selection approach by insertion of an in-frame blasticidin S deaminase marker, we generated a Dd2 reporter line expressing mNeonGreen under the control of the pfpare (P. falciparum Prodrug Activation and Resistance Esterase) locus. We selected the pfpare locus as an integration site because it is highly conserved across P. falciparum strains, expressed throughout the intraerythrocytic cycle, not essential, and offers the potential for negative selection to further enrich for integrants. The mNeonGreen@pare line demonstrates strong fluorescence with a negligible fitness defect. In addition, the construct developed can serve as a tool to fluorescently tag other P. falciparum strains for in vitro experimentation

Influence of Al doping on the critical fields and gap values in magnesium diboride single crystals

The lower ($H_{c1}$) and upper ($H_{c2}$) critical fields of Mg$_{1-x}$Al$_{x}$B$_2$ single crystals (for $x = 0$, 0.1 and $\gtrsim 0.2$) have been deduced from specific heat and local magnetization measurements, respectively. We show that $H_{c1}$ and $H_{c2}$ are both decreasing with increasing doping content. The corresponding anisotropy parameter $\Gamma_{H_{c2}}(0) = H^{ab}_{c2}(0)/H^c_{c2}(0)$ value also decreases from $\sim 5$ in pure MgB$_2$ samples down to $\sim 1.5$ for $x \gtrsim 0.2$ whereas $\Gamma_{H_{c1}}(0)=H^c_{c1}(0)/H^{ab}_{c1}(0)$ remains on the order of 1 in all samples. The small and large gap values have been obtained by fitting the temperature dependence of the zero field electronic contribution to the specific heat to the two gap model for the three Al concentrations. Very similar values have also been obtained by point contact spectroscopy measurements. The evolution of those gaps with Al concentration suggests that both band filling and interband scattering effects are present

Field and angular dependence of the Sommerfeld coefficient in Al-doped MgB2 single crystals

International audienceThe angular and field dependence of the Sommerfeld coefficient = lim Cel /T T→0 Cel being the electronic contribution to the specific heat has been measured in Al-doped MgB2 single crystals for x=0, x=0.1, and x 0.2 . We show that the decomposition previously introduced to describe H, where is the angle between the applied field and the c axis in pure samples Phys. Rev. Lett. 98, 137001 2007 is well adapted to doped samples: e.g., the contribution of the band to the specific heat is proportional to B/Bc2 whereas the contribution of the band is isotropic but highly nonlinear in field. We hence present the evolution of the coherence lengths of the two bands and corresponding Fermi velocities with doping

Anisotropy of the Sommerfeld Coefficient in Magnesium Diboride Single Crystals

International audienceThe anisotropic field dependence of the Sommerfeld coefficient has been measured down to B ! 0 by combining specific heat and Hall probe magnetization measurements in MgB2 single crystals. We find that B; is the sum of two contributions arising from the and band, respectively. We show that B; B=Bc2 where Bc2 Bab c2= si n 2 2cos2 p with 5:4 ( being the angle between the applied field and the c axis) and B; B B=B B . The ''critical field'' of the band B is fully isotropic but field dependent increasing from 0:25 T for B 0:1 T up to 3 T Bcc 2 for B ! 3 T. Because of the coupling of the two bands, superconductivity survives in the band up to 3 T but is totally destroyed above for any orientation of the field

Thermodynamics of Quantum Hall Ferromagnets

The two-dimensional interacting electron gas at Landau level filling factor $\nu =1$ and temperature $T=0$ is a strong ferromagnet; all spins are completely aligned by arbitrarily weak Zeeman coupling. We report on a theoretical study of its thermodynamic properties using a many-body perturbation theory approach and concentrating on the recently measured temperature dependence of the spin magnetization. We discuss the interplay of collective and single-particle aspects of the physics and the opportunities for progress in our understanding of itinerant electron ferromagnetism presented by quantum Hall ferromagnets.Comment: REVTex, 10 pages, 3 uuencoded, compressed and tarred PostScript figures appende

Anisotropies of the lower and upper critical fields in MgB2_2 single crystals

The temperature dependence of the London penetration depth ($\lambda$) and coherence length ($\xi$) has been deduced from Hall probe magnetization measurements in high quality MgB$_2$ single crystals in the two main crystallographic directions. We show that, in contrast to conventional superconductors, MgB$_2$ is characterized by two different anisotropy parameters ($\Gamma_\lambda = \lambda_c/\lambda_{ab}$ and $\Gamma_\xi = \xi_{ab}/\xi_c$) which strongly differ at low temperature and merge at $T_c$. These results are in very good agreement with recent calculations in weakly coupled two bands suprerconductors (Phys. Rev. B, 66, 020509(R) (2002).Comment: 4 pages, 4 figure

CRISPR-mediated genome editing of Plasmodium falciparum malaria parasites

The development of the CRISPR-Cas system is revolutionizing genome editing in a variety of organisms. The system has now been used to manipulate the genome of Plasmodium falciparum, the most lethal malaria-causing species. The ability to generate gene deletions or nucleotide substitutions rapidly and economically promises to accelerate the analysis of novel drug targets and to help elucidate the function of specific genes or gene families, while complementing genome-wide association studies

Machine learning analytics of resting-state functional connectivity predicts survival outcomes of glioblastoma multiforme patients

Glioblastoma multiforme (GBM) is the most frequently occurring brain malignancy. Due to its poor prognosis with currently available treatments, there is a pressing need for easily accessible, non-invasive techniques to help inform pre-treatment planning, patient counseling, and improve outcomes. In this study we determined the feasibility of resting-state functional connectivity (rsFC) to classify GBM patients into short-term and long-term survival groups with respect to reported median survival (14.6 months). We used a support vector machine with rsFC between regions of interest as predictive features. We employed a novel hybrid feature selection method whereby features were first filtered using correlations between rsFC and OS, and then using the established method of recursive feature elimination (RFE) to select the optimal feature subset. Leave-one-subject-out cross-validation evaluated the performance of models. Classification between short- and long-term survival accuracy was 71.9%. Sensitivity and specificity were 77.1 and 65.5%, respectively. The area under the receiver operating characteristic curve was 0.752 (95% CI, 0.62-0.88). These findings suggest that highly specific features of rsFC may predict GBM survival. Taken together, the findings of this study support that resting-state fMRI and machine learning analytics could enable a radiomic biomarker for GBM, augmenting care and planning for individual patients

Randomized trial of conventional transseptal needle versus radiofrequency energy needle puncture for left atrial access (the TRAVERSE-LA study).

BackgroundTransseptal puncture is a critical step in achieving left atrial (LA) access for a variety of cardiac procedures. Although the mechanical Brockenbrough needle has historically been used for this procedure, a needle employing radiofrequency (RF) energy has more recently been approved for clinical use. We sought to investigate the comparative effectiveness of an RF versus conventional needle for transseptal LA access.Methods and resultsIn this prospective, single-blinded, controlled trial, 72 patients were randomized in a 1:1 fashion to an RF versus conventional (BRK-1) transseptal needle. In an intention-to-treat analysis, the primary outcome was time required for transseptal LA access. Secondary outcomes included failure of the assigned needle, visible plastic dilator shavings from needle introduction, and any procedural complication. The median transseptal puncture time was 68% shorter using the RF needle compared with the conventional needle (2.3 minutes [interquartile range {IQR}, 1.7 to 3.8 minutes] versus 7.3 minutes [IQR, 2.7 to 14.1 minutes], P = 0.005). Failure to achieve transseptal LA access with the assigned needle was less common using the RF versus conventional needle (0/36 [0%] versus 10/36 [27.8%], P &lt; 0.001). Plastic shavings were grossly visible after needle advancement through the dilator and sheath in 0 (0%) RF needle cases and 12 (33.3%) conventional needle cases (P &lt; 0.001). There were no differences in procedural complications (1/36 [2.8%] versus 1/36 [2.8%]).ConclusionsUse of an RF needle resulted in shorter time to transseptal LA access, less failure in achieving transseptal LA access, and fewer visible plastic shavings

Historical geography II: traces remain

The second report in this series turns to focus on the trace in relation to life-writing and biography in historical geography and beyond. Through attention to tracing journeys, located moments and listening to the presence of ghosts (Ogborn, 2005), this report seeks to highlight the range of different ways in which historical geographers have explored lives, deaths, and their transient traces through varied biographical terrains. Continuing to draw attention in historical geography to the darkest of histories, this piece will pivot on moments of discovering the dead to showcase the nuanced ways in which historical geography is opening doors into uncharted lives and unspoken histories