The use of Multiple Displacement Amplification to increase the detection and genotyping of <i>Trypanosoma</i> samples immobilised on FTA filters.

Whole genome amplification methods are a recently developed tool for amplifying DNA from limited template. We report its application in trypanosome infections, characterized by low parasitemias. Multiple displacement amplification (MDA) amplifies DNA with a simple in vitro step and was evaluated on mouse blood samples on FTA filter cards with known numbers of Trypanosoma brucei parasites. The data showed a 20-fold increase in the number of PCRs possible per sample, using primers diagnostic for the multicopy ribosomal ITS region or 177-bp repeats, and a 20-fold increase in sensitivity over nested PCR against a single-copy microsatellite. Using MDA for microsatellite genotyping caused allele dropout at low DNA concentrations, which was overcome by pooling multiple MDA reactions. The validity of using MDA was established with samples from Human African Trypanosomiasis patients. The use of MDA allows maximal use of finite DNA samples and may prove a valuable tool in studies where multiple reactions are necessary, such as population genetic analyses

Weyl asymptotics for magnetic Schr\"odinger operators and de Gennes' boundary condition

This paper is concerned with the discrete spectrum of the self-adjoint realization of the semi-classical Schr\"odinger operator with constant magnetic field and associated with the de Gennes (Fourier/Robin) boundary condition. We derive an asymptotic expansion of the number of eigenvalues below the essential spectrum (Weyl-type asymptotics). The methods of proof relies on results concerning the asymptotic behavior of the first eigenvalue obtained in a previous work [A. Kachmar, J. Math. Phys. Vol. 47 (7) 072106 (2006)].Comment: 28 pages (revised version). to appear in Rev Math Phy

Getting a handle on lipid droplets: Insights into ER-lipid droplet tethering.

Lipid droplets (LDs) are hubs for lipid metabolism that form membrane contact sites with multiple organelles. In this issue, Hariri et al. (2019. J. Cell Biol. https://doi.org/10.1083/jcb.201808119) reveal the functions of Mdm1-mediated endoplasmic reticulum (ER)-LD tethering in yeast and Datta et al. (2019. J. Cell Biol. https://doi.org/10.1083/jcb.201808133) identify a role for the Mdm1 orthologue, Snx14, as an ER-LD tether that regulates lipid metabolism in human cells

Effect of RpoN, RpoS and LuxS Pathways on the Biofilm Formation and Antibiotic Sensitivity of Borrelia burgdorferi

Borrelia burgdorferi, the causative agent of Lyme disease, is capable of forming biofilm in vivo and in vitro, a structure well known for its resistance to antimicrobial agents. For the formation of biofilm, signaling processes are required to communicate with the surrounding environment such as it was shown for the RpoN—RpoS alternative sigma factor and for the LuxS quorum-sensing pathways. Therefore, in this study, the wild-type B. burgdorferi and different mutant strains lacking RpoN, RpoS, and LuxS genes were studied for their growth characteristic and development of biofilm structures and markers as well as for their antibiotic sensitivity. Our results showed that all three mutants formed small, loosely formed aggregates, which expressed previously identified Borrelia biofilm markers such as alginate, extracellular DNA, and calcium. All three mutants had significantly different sensitivity to doxycyline in the early log phase spirochete cultures; however, in the biofilm rich stationary cultures, only LuxS mutant showed increased sensitivity to doxycyline compared to the wild-type strain. Our findings indicate that all three mutants have some effect on Borrelia biofilm, but the most dramatic effect was found with LuxS mutant, suggesting that the quorum-sensing pathway plays an important role of Borrelia biofilm formation and antibiotic sensitivity

Eigenvalues of Laplacian with constant magnetic field on non-compact hyperbolic surfaces with finite area

We consider a magnetic Laplacian $-\Delta_A=(id+A)^\star (id+A)$ on a noncompact hyperbolic surface \mM with finite area. $A$ is a real one-form and the magnetic field $dA$ is constant in each cusp. When the harmonic component of $A$ satifies some quantified condition, the spectrum of $-\Delta_A$ is discrete. In this case we prove that the counting function of the eigenvalues of $-\Delta_{A}$ satisfies the classical Weyl formula, even when $dA=0.

Comprehensive Phylogenetic Reconstruction of Amoebozoa Based on Concatenated Analyses of SSU-rDNA and Actin Genes

Evolutionary relationships within Amoebozoa have been the subject of controversy for two reasons: 1) paucity of morphological characters in traditional surveys and 2) haphazard taxonomic sampling in modern molecular reconstructions. These along with other factors have prevented the erection of a definitive system that resolves confidently both higher and lower-level relationships. Additionally, the recent recognition that many protosteloid amoebae are in fact scattered throughout the Amoebozoa suggests that phylogenetic reconstructions have been excluding an extensive and integral group of organisms. Here we provide a comprehensive phylogenetic reconstruction based on 139 taxa using molecular information from both SSU-rDNA and actin genes. We provide molecular data for 13 of those taxa, 12 of which had not been previously characterized. We explored the dataset extensively by generating 18 alternative reconstructions that assess the effect of missing data, long-branched taxa, unstable taxa, fast evolving sites and inclusion of environmental sequences. We compared reconstructions with each other as well as against previously published phylogenies. Our analyses show that many of the morphologically established lower-level relationships (defined here as relationships roughly equivalent to Order level or below) are congruent with molecular data. However, the data are insufficient to corroborate or reject the large majority of proposed higher-level relationships (above the Order-level), with the exception of Tubulinea, Archamoebae and Myxogastrea, which are consistently recovered. Moreover, contrary to previous expectations, the inclusion of available environmental sequences does not significantly improve the Amoebozoa reconstruction. This is probably because key amoebozoan taxa are not easily amplified by environmental sequencing methodology due to high rates of molecular evolution and regular occurrence of large indels and introns. Finally, in an effort to facilitate future sampling of key amoebozoan taxa, we provide a novel methodology for genome amplification and cDNA extraction from single or a few cells, a method that is culture-independent and allows both photodocumentation and extraction of multiple genes from natural samples

Molecular characterization of Geitleria appalachiana sp. nov. (Nostocales, Cyanobacteria) and formation of Geitleriaceae fam. nov.

Geitleria was described from a limestone cave in Israel, and subsequently reported from caves of France, Romania, Spain, Florida, Costa Rica, and Cook Islands. It is morphologically unusual in that it has true branching, but no heterocytes. A morphologically distinct species of Geitleria was recently collected from a limestone cave in Great Smoky Mountains National Park, Tennessee, and is herein described as G. appalachiana sp. nov. Sequence data for 16S rRNA and rpoC1 loci for the species were obtained from field material using single filament PCR. Phylogenetic evidence indicates that Geitleria does not belong to any family in the Nostocales containing true–branching genera, i.e. Hapalosiphonaceae, Chlorogloeopsidaceae, and Symphyonemataceae, and consequently Geitleriaceae fam. nov. is established to contain this unique genus

Essential self-adjointness for combinatorial Schr\"odinger operators II- Metrically non complete graphs

We consider weighted graphs, we equip them with a metric structure given by a weighted distance, and we discuss essential self-adjointness for weighted graph Laplacians and Schr\"odinger operators in the metrically non complete case.Comment: Revisited version: Ognjen Milatovic wrote to us that he had discovered a gap in the proof of theorem 4.2 of our paper. As a consequence we propose to make an additional assumption (regularity property of the graph) to this theorem. A new subsection (4.1) is devoted to the study of this property and some details have been changed in the proof of theorem 4.

Excess TGF-β Induces MMP-9 and SPP-1 Expression in the Skeletal Muscle of Cancer Patients with Bone Metastases: Association with Muscle Dysfunction

https://openworks.mdanderson.org/sumexp23/1052/thumbnail.jp