Droplet-based microfluidic screening and sorting of microalgal populations for strain engineering applications

The application of microfluidic technologies to microalgal research is particularly appealing since these approaches allow the precise control of the extracellular environment and offer a high-throughput approach to studying dynamic cellular processes. To expand the portfolio of applications, here we present a droplet-based microfluidic method for analysis and screening of Phaeodactylum tricornutum and Nannochloropsis gaditana, which can be integrated into a genetic transformation workflow. Following encapsulation of single cells in picolitre-sized droplets, fluorescence signals arising from each cell can be used to assess its phenotypic state. In this work, the chlorophyll fluorescence intensity of each cell was quantified and used to identify populations of P. tricornutum cells grown in different light conditions. Further, individual P. tricornutum or N. gaditana cells engineered to express green fluorescent protein were distinguished and sorted from wild-type cells. This has been exploited as a rapid screen for transformed cells within a population, bypassing a major bottleneck in algal transformation workflows and offering an alternative strategy for the identification of genetically modified strains

The bmi-1 oncoprotein is differentially expressed in non-small cell lung cancer and correlates with INK4A-ARF locus expression

Genes of the polycomb group function by silencing homeotic selector genes that regulate embryogenesis. In mice, downregulation of one of the polycomb genes, bmi-1, leads to neurological alterations and severe proliferative defects in lymphoid cells, whilst bmi-1 overexpression, together with upregulation of myc-1, induces lymphoma. An oncogenic function has been further supported in primary fibroblast studies where bmi-1 overexpression induces immortalization due to repression of p16/p19ARF, and where together with H-ras, it readily transforms MEFs. It was the aim of this study to assess the expression of bmi-1 in resectable non-small cell lung cancer (NSCLC) in association with p16 and p14ARF (=human p19ARF). Tumours (48 resectable NSCLC (32 squamous, 9 adeno-, 2 large cell, 4 undifferentiated carcinomas and 1 carcinoid); stage I, 29, II, 7, III, 12; T1, 18, T2, 30; differentiation: G1 12, G2 19, G3 17) were studied by immunohistochemistry for protein expression and by comparative multiplex PCR for gene amplification analysis. In tumour-free, normal lung tissue from patients, weak – moderate bmi-1 staining was seen in some epithelial cells, lymphocytes, glandular cells and in fibroblasts, whereas blood, endothelial, chondrocytes, muscle cells and adipocytes did not exhibit any bmi-1 expression. In tumours, malignant cells were negative/weakly, moderately and strongly positive in 20, 22 and 6 cases, respectively. As assessed by multiplex PCR, bmi-1 gene amplification was not the reason for high-level bmi-1 expression. Tumours with moderate or strong bmi-1 expression were more likely to have low levels of p16 and p14ARF (P = 0.02). Similarly, tumours negative for both, p16 and p14ARF, exhibit moderate–strong bmi-1 staining. 58% of resectable NSCLC exhibit moderate–high levels of bmi-1 protein. The inverse correlation of bmi-1 and the INK4 locus proteins expression (p16/p14ARF) supports a possible role for bmi-1 misregulation in lung carcinogenesis. © 2001 Cancer Research Campaign www.bjcancer.co

Weak Ferromagnetism and Excitonic Condensates

We investigate a model of excitonic ordering (i.e electron-hole pair condensation) appropriate for the divalent hexaborides. We show that the inclusion of imperfectly nested electron hole Fermi surfaces can lead to the formation of an undoped excitonic metal phase. In addition, we find that weak ferromagnetism with compensated moments arises as a result of gapless excitations. We study the effect of the low lying excitations on the density of states, Fermi surface topology and optical conductivity and compare to available experimental data.Comment: 10 Pages, 8 Figures, RevTe

Point defects, ferromagnetism and transport in calcium hexaboride

The formation energy and local magnetic moment of a series of point defects in CaB$_6$ are computed using a supercell approach within the generalized gradient approximation to density functional theory. Based on these results, speculations are made as to the influence of these defects on electrical transport. It is found that the substitution of Ca by La does not lead to the formation of a local moment, while a neutral B$_6$ vacancy carries a moment of 2.4 Bohr magnetons, mostly distributed over the six nearest-neighbour B atoms. A plausible mechanism for the ferromagnetic ordering of these moments is suggested. Since the same broken B-B bonds appear on the preferred (100) cleavage planes of the CaB$_6$ structure, it is argued that internal surfaces in polycrystals as well as external surfaces in general will make a large contribution to the observed magnetization.Comment: Calculated defect formation energies had to be corrected, due to the use of a wrong reference energy for the perfect crystal in the original pape

Cosmesis and body image after single-port laparoscopic or conventional laparoscopic cholecystectomy: a multicenter double blinded randomised controlled trial (SPOCC-trial)

BACKGROUND: Emerging attempts have been made to reduce operative trauma and improve cosmetic results of laparoscopic cholecystectomy. There is a trend towards minimizing the number of incisions such as natural transluminal endoscopic surgery (NOTES) and single-port laparoscopic cholecystectomy (SPLC). Many retrospective case series propose excellent cosmesis and reduced pain in SPLC. As the latter has been confirmed in a randomized controlled trial, patient's satisfaction on cosmesis is still controversially debated. METHODS/DESIGN: The SPOCC trial is a prospective, multi-center, double blinded, randomized controlled study comparing SPLC with 4-port conventional laparoscopic cholecystectomy (4PLC) in elective surgery. The hypothesis and primary objective is that patients undergoing SPLC will have a better outcome in cosmesis and body image 12 weeks after surgery. This primary endpoint is assessed using a validated 8-item multiple choice type questionnaire on cosmesis and body image. The secondary endpoint has three entities: the quality of life 12 weeks after surgery assessed by the validated Short-Form-36 Health Survey questionnaire, postoperative pain assessed by a visual analogue scale and the use of analgesics. Operative time, surgeon's experience with SPLC and 4PLC, use of additional ports, conversion to 4PLC or open cholecystectomy, length of stay, costs, time of work as well as intra- and postoperative complications are further aspects of the secondary endpoint. Patients are randomly assigned either to SPLC or to 4PLC. Patients as well as treating physicians, nurses and assessors are blinded until the 7th postoperative day. Sample size calculation performed by estimating a difference of cosmesis of 20% (alpha = 0.05 and beta = 0.90, drop out rate of 10%) resulted in a number of 55 randomized patients per arm. DISCUSSION: The SPOCC-trial is a prospective, multi-center, double-blind, randomized controlled study to assess cosmesis and body image after SPLC

The Cosmic Microwave Background in an Inhomogeneous Universe - why void models of dark energy are only weakly constrained by the CMB

The dimming of Type Ia supernovae could be the result of Hubble-scale inhomogeneity in the matter and spatial curvature, rather than signaling the presence of a dark energy component. A key challenge for such models is to fit the detailed spectrum of the cosmic microwave background (CMB). We present a detailed discussion of the small-scale CMB in an inhomogeneous universe, focusing on spherically symmetric `void' models. We allow for the dynamical effects of radiation while analyzing the problem, in contrast to other work which inadvertently fine tunes its spatial profile. This is a surprisingly important effect and we reach substantially different conclusions. Models which are open at CMB distances fit the CMB power spectrum without fine tuning; these models also fit the supernovae and local Hubble rate data which favours a high expansion rate. Asymptotically flat models may fit the CMB, but require some extra assumptions. We argue that a full treatment of the radiation in these models is necessary if we are to understand the correct constraints from the CMB, as well as other observations which rely on it, such as spectral distortions of the black body spectrum, the kinematic Sunyaev-Zeldovich effect or the Baryon Acoustic Oscillations.Comment: 23 pages with 14 figures. v2 has considerably extended discussion and analysis, but the basic results are unchanged. v3 is the final versio

High Magnetic Field NMR Studies of LiVGe2_2O6_6, a quasi 1-D Spin S=1S = 1 System

We report $^{7}$Li pulsed NMR measurements in polycrystalline and single crystal samples of the quasi one-dimensional S=1 antiferromagnet LiVGe$_2$O$_6$, whose AF transition temperature is $T_{\text{N}}\simeq 24.5$ K. The field ($B_0$) and temperature ($T$) ranges covered were 9-44.5 T and 1.7-300 K respectively. The measurements included NMR spectra, the spin-lattice relaxation rate ($T_1^{-1}$), and the spin-phase relaxation rate ($T_2^{-1}$), often as a function of the orientation of the field relative to the crystal axes. The spectra indicate an AF magnetic structure consistent with that obtained from neutron diffraction measurements, but with the moments aligned parallel to the c-axis. The spectra also provide the $T$-dependence of the AF order parameter and show that the transition is either second order or weakly first order. Both the spectra and the $T_1^{-1}$ data show that $B_0$ has at most a small effect on the alignment of the AF moment. There is no spin-flop transition up to 44.5 T. These features indicate a very large magnetic anisotropy energy in LiVGe$_2$O$_6$ with orbital degrees of freedom playing an important role. Below 8 K, $T_1^{-1}$ varies substantially with the orientation of $B_0$ in the plane perpendicular to the c-axis, suggesting a small energy gap for magnetic fluctuations that is very anisotropic.Comment: submitted to Phys. Rev.

CaB_6: a new semiconducting material for spin electronics

Ferromagnetism was recently observed at unexpectedly high temperatures in La-doped CaB_6. The starting point of all theoretical proposals to explain this observation is a semimetallic electronic structure calculated for CaB_6 within the local density approximation. Here we report the results of parameter-free quasiparticle calculations of the single-particle excitation spectrum which show that CaB_6 is not a semimetal but a semiconductor with a band gap of 0.8 eV. Magnetism in La_xCa_{1-x}B_6 occurs just on the metallic side of a Mott transition in the La-induced impurity band.Comment: 4 pages, 1 postscript figur

Chemistry of heavy elements in the Dark Ages

Primordial molecules were formed during the Dark Ages, i.e. the time between recombination and reionization in the early Universe. The purpose of this article is to analyze the formation of primordial molecules based on heavy elements during the Dark Ages, with elemental abundances taken from different nucleosynthesis models. We present calculations of the full non-linear equation set governing the primordial chemistry. We consider the evolution of 45 chemical species and use an implicit multistep method of variable order of precision with an adaptive stepsize control. We find that the most abundant Dark Ages molecules based on heavy elements are CH and OH. Non-standard nucleosynthesis can lead to higher heavy element abundances while still satisfying the observed primordial light abundances. In that case, we show that the abundances of molecular species based on C, N, O and F can be enhanced by two orders of magnitude compared to the standard case, leading to a CH relative abundance higher than that of HD+ or H2D+.Comment: 14 pages, accepted by Astronomy and Astrophysic

The first spectral line surveys searching for signals from the Dark Ages

Our aim is to observationally investigate the cosmic Dark Ages in order to constrain star and structure formation models, as well as the chemical evolution in the early Universe. Spectral lines from atoms and molecules in primordial perturbations at high redshifts can give information about the conditions in the early universe before and during the formation of the first stars in addition to the epoch of reionisation. The lines may arise from moving primordial perturbations before the formation of the first stars (resonant scattering lines), or could be thermal absorption or emission lines at lower redshifts. The difficulties in these searches are that the source redshift and evolutionary state, as well as molecular species and transition are unknown, which implies that an observed line can fall within a wide range of frequencies. The lines are also expected to be very weak. Observations from space have the advantages of stability and the lack of atmospheric features which is important in such observations. We have therefore, as a first step in our searches, used the Odin satellite to perform two sets of spectral line surveys towards several positions. The first survey covered the band 547-578 GHz towards two positions, and the second one covered the bands 542.0-547.5 GHz and 486.5-492.0 GHz towards six positions selected to test different sizes of the primordial clouds. Two deep searches centred at 543.250 and 543.100 GHz with 1 GHz bandwidth were also performed towards one position. The two lowest rotational transitions of H2 will be redshifted to these frequencies from z~20-30, which is the predicted epoch of the first star formation. No lines are detected at an rms level of 14-90 and 5-35 mK for the two surveys, respectively, and 2-7 mK in the deep searches with a channel spacing of 1-16 MHz. The broad bandwidth covered allows a wide range of redshifts to be explored for a number of atomic and molecular species and transitions. From the theoretical side, our sensitivity analysis show that the largest possible amplitudes of the resonant lines are about 1 mK at frequencies <200 GHz, and a few micro K around 500-600 GHz, assuming optically thick lines and no beam-dilution. However, if existing, thermal absorption lines have the potential to be orders of magnitude stronger than the resonant lines. We make a simple estimation of the sizes and masses of the primordial perturbations at their turn-around epochs, which previously has been identified as the most favourable epoch for a detection. This work may be considered as an important pilot study for our forthcoming observations with the Herschel Space Observatory.Comment: 15 pages, 9 figures, 3 on-line pages. Accepted for publication in Astronomy & Astrophysics 8 March 2010