Application of magnetically induced hyperthermia on the model protozoan Crithidia fasciculata as a potential therapy against parasitic infections

Magnetic hyperthermia is currently an EU-approved clinical therapy against tumor cells that uses magnetic nanoparticles under a time varying magnetic field (TVMF). The same basic principle seems promising against trypanosomatids causing Chagas disease and sleeping sickness, since therapeutic drugs available display severe side effects and drug-resistant strains. However, no applications of this strategy against protozoan-induced diseases have been reported so far. In the present study, Crithidia fasciculata, a widely used model for therapeutic strategies against pathogenic trypanosomatids, was targeted with Fe_{3}O_{4} magnetic nanoparticles (MNPs) in order to remotely provoke cell death using TVMFs. The MNPs with average sizes of d approx. 30 nm were synthesized using a precipitation of FeSO_{4}4 in basic medium. The MNPs were added to Crithidia fasciculata choanomastigotes in exponential phase and incubated overnight. The amount of uploaded MNPs per cell was determined by magnetic measurements. Cell viability using the MTT colorimetric assay and flow cytometry showed that the MNPs were incorporated by the cells with no noticeable cell-toxicity effects. When a TVMF (f = 249 kHz, H = 13 kA/m) was applied to MNP-bearing cells, massive cell death was induced via a non-apoptotic mechanism. No effects were observed by applying a TVMF on control (without loaded MNPs) cells. No macroscopic rise in temperature was observed in the extracellular medium during the experiments. Scanning Electron Microscopy showed morphological changes after TVMF experiments. These data indicate (as a proof of principle) that intracellular hyperthermia is a suitable technology to induce the specific death of protozoan parasites bearing MNPs. These findings expand the possibilities for new therapeutic strategies that combat parasitic infections.Comment: 9 pages, four supplementary video file

Self-renewal of single mouse hematopoietic stem cells is reduced by JAK2V617F without compromising progenitor cell expansion

Recent descriptions of significant heterogeneity in normal stem cells and cancers have altered our understanding of tumorigenesis, emphasizing the need to understand how single stem cells are subverted to cause tumors. Human myeloproliferative neoplasms (MPNs) are thought to reflect transformation of a hematopoietic stem cell (HSC) and the majority harbor an acquired V617F mutation in the JAK2 tyrosine kinase, making them a paradigm for studying the early stages of tumor establishment and progression. The consequences of activating tyrosine kinase mutations for stem and progenitor cell behavior are unclear. In this article, we identify a distinct cellular mechanism operative in stem cells. By using conditional knock-in mice, we show that the HSC defect resulting from expression of heterozygous human JAK2V617F is both quantitative (reduced HSC numbers) and qualitative (lineage biases and reduced self-renewal per HSC). The defect is intrinsic to individual HSCs and their progeny are skewed toward proliferation and differentiation as evidenced by single cell and transplantation assays. Aged JAK2V617F show a more pronounced defect as assessed by transplantation, but mice that transform reacquire competitive self-renewal ability. Quantitative analysis of HSC-derived clones was used to model the fate choices of normal and JAK2-mutant HSCs and indicates that JAK2V617F reduces self-renewal of individual HSCs but leaves progenitor expansion intact. This conclusion is supported by paired daughter cell analyses, which indicate that JAK2-mutant HSCs more often give rise to two differentiated daughter cells. Together these data suggest that acquisition of JAK2V617F alone is insufficient for clonal expansion and disease progression and causes eventual HSC exhaustion. Moreover, our results show that clonal expansion of progenitor cells provides a window in which collaborating mutations can accumulate to drive disease progression. Characterizing the mechanism(s) of JAK2V617F subclinical clonal expansions and the transition to overt MPNs will illuminate the earliest stages of tumor establishment and subclone competition, fundamentally shifting the way we treat and manage cancers

Bifurcations of periodic orbits with spatio-temporal symmetries

Motivated by recent analytical and numerical work on two- and three-dimensional convection with imposed spatial periodicity, we analyse three examples of bifurcations from a continuous group orbit of spatio-temporally symmetric periodic solutions of partial differential equations. Our approach is based on centre manifold reduction for maps, and is in the spirit of earlier work by Iooss (1986) on bifurcations of group orbits of spatially symmetric equilibria. Two examples, two-dimensional pulsating waves (PW) and three-dimensional alternating pulsating waves (APW), have discrete spatio-temporal symmetries characterized by the cyclic groups Z_n, n=2 (PW) and n=4 (APW). These symmetries force the Poincare' return map M to be the nth iterate of a map G: M=G^n. The group orbits of PW and APW are generated by translations in the horizontal directions and correspond to a circle and a two-torus, respectively. An instability of pulsating waves can lead to solutions that drift along the group orbit, while bifurcations with Floquet multiplier +1 of alternating pulsating waves do not lead to drifting solutions. The third example we consider, alternating rolls, has the spatio-temporal symmetry of alternating pulsating waves as well as being invariant under reflections in two vertical planes. This leads to the possibility of a doubling of the marginal Floquet multiplier and of bifurcation to two distinct types of drifting solutions. We conclude by proposing a systematic way of analysing steady-state bifurcations of periodic orbits with discrete spatio-temporal symmetries, based on applying the equivariant branching lemma to the irreducible representations of the spatio-temporal symmetry group of the periodic orbit, and on the normal form results of Lamb (1996). This general approach is relevant to other pattern formation problems, and contributes to our understanding of the transition from ordered to disordered behaviour in pattern-forming systems

Faraday waves on a viscoelastic liquid

We investigate Faraday waves on a viscoelastic liquid. Onset measurements and a nonlinear phase diagram for the selected patterns are presented. By virtue of the elasticity of the material a surface resonance synchronous to the external drive competes with the usual subharmonic Faraday instability. Close to the bicriticality the nonlinear wave interaction gives rise to a variety of novel surface states: Localised patches of hexagons, hexagonal superlattices, coexistence of hexagons and lines. Theoretical stability calculations and qualitative resonance arguments support the experimental observations.Comment: 4 pages, 4figure

Pattern formation in 2-frequency forced parametric waves

We present an experimental investigation of superlattice patterns generated on the surface of a fluid via parametric forcing with 2 commensurate frequencies. The spatio-temporal behavior of 4 qualitatively different types of superlattice patterns is described in detail. These states are generated via a number of different 3--wave resonant interactions. They occur either as symmetry--breaking bifurcations of hexagonal patterns composed of a single unstable mode or via nonlinear interactions between the two primary unstable modes generated by the two forcing frequencies. A coherent picture of these states together with the phase space in which they appear is presented. In addition, we describe a number of new superlattice states generated by 4--wave interactions that arise when symmetry constraints rule out 3--wave resonances.Comment: The paper contains 34 pages and 53 figures and provides an extensive review of both the theoretical and experimental work peformed in this syste

Isodicentric Y Chromosomes and Sex Disorders as Byproducts of Homologous Recombination that Maintains Palindromes

Massive palindromes in the human Y chromosome harbor mirror-image gene pairs essential for spermatogenesis. During evolution, these gene pairs have been maintained by intrapalindrome, arm-to-arm recombination. The mechanism of intrapalindrome recombination and risk of harmful effects are unknown. We report 51 patients with isodicentric Y (idicY) chromosomes formed by homologous crossing over between opposing arms of palindromes on sister chromatids. These ectopic recombination events occur at nearly all Y-linked palindromes. Based on our findings, we propose that intrapalindrome sequence identity is maintained via noncrossover pathways of homologous recombination. DNA double-strand breaks that initiate these pathways can be alternatively resolved by crossing over between sister chromatids to form idicY chromosomes, with clinical consequences ranging from spermatogenic failure to sex reversal and Turner syndrome. Our observations imply that crossover and noncrossover pathways are active in nearly all Y-linked palindromes, exposing an Achilles' heel in the mechanism that preserves palindrome-borne genes.National Institutes of Health (U.S.)Howard Hughes Medical InstituteNetherlands Organization for Scientific ResearchUniversity of Amsterdam. Academic Medical CenterBoehringer Ingelheim (Fellowship

ProtoDESI: First On-Sky Technology Demonstration for the Dark Energy Spectroscopic Instrument

The Dark Energy Spectroscopic Instrument (DESI) is under construction to measure the expansion history of the universe using the baryon acoustic oscillations technique. The spectra of 35 million galaxies and quasars over 14,000 square degrees will be measured during a 5-year survey. A new prime focus corrector for the Mayall telescope at Kitt Peak National Observatory will deliver light to 5,000 individually targeted fiber-fed robotic positioners. The fibers in turn feed ten broadband multi-object spectrographs. We describe the ProtoDESI experiment, that was installed and commissioned on the 4-m Mayall telescope from August 14 to September 30, 2016. ProtoDESI was an on-sky technology demonstration with the goal to reduce technical risks associated with aligning optical fibers with targets using robotic fiber positioners and maintaining the stability required to operate DESI. The ProtoDESI prime focus instrument, consisting of three fiber positioners, illuminated fiducials, and a guide camera, was installed behind the existing Mosaic corrector on the Mayall telescope. A Fiber View Camera was mounted in the Cassegrain cage of the telescope and provided feedback metrology for positioning the fibers. ProtoDESI also provided a platform for early integration of hardware with the DESI Instrument Control System that controls the subsystems, provides communication with the Telescope Control System, and collects instrument telemetry data. Lacking a spectrograph, ProtoDESI monitored the output of the fibers using a Fiber Photometry Camera mounted on the prime focus instrument. ProtoDESI was successful in acquiring targets with the robotically positioned fibers and demonstrated that the DESI guiding requirements can be met.Comment: Accepted versio

Rapid sympathetic cooling to Fermi degeneracy on a chip

Neutral fermions present new opportunities for testing many-body condensed matter systems, realizing precision atom interferometry, producing ultra-cold molecules, and investigating fundamental forces. However, since their first observation, quantum degenerate Fermi gases (DFGs) have continued to be challenging to produce, and have been realized in only a handful of laboratories. In this Letter, we report the production of a DFG using a simple apparatus based on a microfabricated magnetic trap. Similar approaches applied to Bose-Einstein Condensation (BEC) of 87Rb have accelerated evaporative cooling and eliminated the need for multiple vacuum chambers. We demonstrate sympathetic cooling for the first time in a microtrap, and cool 40K to Fermi degeneracy in just six seconds -- faster than has been possible in conventional magnetic traps. To understand our sympathetic cooling trajectory, we measure the temperature dependence of the 40K-87Rb cross-section and observe its Ramsauer-Townsend reduction.Comment: 5 pages, 4 figures (v3: new collision data, improved atom number calibration, revised text, improved figures.

Association of British Neurologists: revised (2015) guidelines for prescribing disease-modifying treatments in multiple sclerosis.

In June 1999, the Association of British Neurologists (ABN) first published guidelines for the use of the licensed multiple sclerosis (MS) disease-modifying treatments (at that time β-interferon and glatiramer acetate). The guidelines were revised in 2001 and have been periodically updated since then. In 2002, following the negative assessment of these treatments by the National Institute for Health and Care Excellence (NICE), the MS risk-sharing scheme started, in which patients eligible according to the 2001 ABN guidelines were provided with treatment funded through the UK National Health Service (NHS), and monitored annually for up to 10 years.1 Recruitment to the risk-sharing scheme cohort is complete. Pending a future final evaluation, the UK Department of Health's instruction to NHS funders remains in place: that patients who fulfil the ABN criteria should continue to receive treatment funded through the NHS. The British neurological community has fully accepted the risk-sharing scheme for prescribing β-interferon and glatiramer acetate. Approximately 70 ‘treating centres’ have recruited >5000 patients between 2002 and 2005, and these have been monitored annually for 10 years; many more patients have received these treatments since 2005. The ABN published revised guidelines in 2007, and then again in 2009, following the licensing of natalizumab and mitoxantrone. This 2015 revised guideline replaces former versions. It includes all newly approved or licensed treatments for MS and represents a consensus concerning their use. These guidelines will require future revision as other treatments receive approval (eg, daclizumab and ocrelizumab): we suggest they are reviewed after an interval of no longer than 12 months. The guideline is not intended to provide a complete description of the possible complications and monitoring of disease-modifying treatments in MS; we refer prescribing neurologists to the relevant summaries of product characteristics.PostprintPeer reviewe