Comparison of the shaping ability of RaCe, FlexMaster, and ProFile nickel-titanium instruments in severely curved root canals

This in vitro study compared the shaping ability of RaCe, FlexMaster, and ProFile rotary nickel-titanium instruments in severely curved root canals of extracted teeth. Sixty maxillary molars with curvatures ranging from 25° to 65° were embedded in a muffle system and portioned into five horizontal sections (thickness 1.2 mm), starting from the apex. Canals were divided into three groups (n = 20, each) and were prepared with RaCe, FlexMaster, or ProFile rotary nickel-titanium instruments and the TriAuto ZX handpiece using a crown-down preparation technique. We evaluated the difference between pre- and postoperative root canal cross-sections, loss of working length, instrument failure, and preparation time. The root canal area before and after the intervention was determined using an area-measuring software. The data were analyzed statistically using a one-way ANOVA followed by a Kruskal-Wallis multiple-comparison Z-value test. Specimens treated with FlexMaster showed the greatest change from preoperative cross-sections, followed by RaCe and ProFile. The cross-sectional changes induced by RaCe and FlexMaster preparation differed significantly from those produced by ProFile. Loss of working length, instrument failure, and preparation time did not differ significantly between the groups. Root canal preparation with the three instruments did not lead to any significant alteration of the original root anatomy or working length. Thus, we conclude that RaCe, FlexMaster, and ProFile instruments are of comparable efficiency and usefulness in the preparation of severely curved root canals

Diagnostics-in-a-Suitcase: Development of a portable and rapid assay for the detection of the emerging avian influenza A (H7N9) virus

Background: In developing countries, the necessary equipment for the diagnosis is only available in few central laboratories, which are less accessible and of limited capacity to test large numbers of incoming samples. Moreover, transport conditions of samples are inadequate and therefore lead to unreliable results. Objectives: The development of rapid, inexpensive, and simple test would allow mobile detection of viruses. Study Design: A suitcase laboratory “Diagnostics-in-a-Suitcase” (56 × 45.5 × 26.5 cm) containing all necessary reagents and devices to perform reverse transcription recombinase polymerase amplification (RT-RPA) assay was developed. As an example, Two RT-RPA assays for the detection of hemagglutinin (H) and neuraminidase (N) genes of the novel avian influenza (H7N9) virus were established. Results: Sensitivities were 10 and 100 \{RNA\} molecules for the \{H7\} and the \{N9\} RT-RPA assays, respectively. Assays were performed at a single temperature (42 °C). Results were obtained within 2-7 minutes. The \{H7N9\} RT-RPA assays showed neither a cross-detection of any other respiratory viruses affecting humans and/or birds nor of the human or chicken genomes. All reagents were used, stored, and transported at ambient temperature, i.e. cold chain independent. In addition, the Diagnostics-in-a-Suitcase was operated by a solar power battery. Conclusions: The developed assay protocol and mobile setup performed well. Moreover, it can be easily implemented to perform diagnosis at airport, quarantine stations, or farms for rapid on-site viral nucleic acid detection

Reverse transcription recombinase polymerase amplification assay for the detection of middle East respiratory syndrome coronavirus

The emergence of Middle East Respiratory Syndrome Coronavirus (MERS-CoV) in the eastern Mediterranean and imported cases to Europe has alerted public health authorities. Currently, detection of MERS-CoV in patient samples is done by real-time RT-PCR. Samples collected from suspected cases are sent to highly-equipped centralized laboratories for screening. A rapid point-of-care test is needed to allow more widespread mobile detection of the virus directly from patient material. In this study, we describe the development of a reverse transcription isothermal Recombinase Polymerase Amplification (RT-RPA) assay for the identification of MERS-CoV. A partial nucleocapsid gene RNA molecular standard of MERS-coronavirus was used to determine the assay sensitivity. The isothermal (42°C) MERS-CoV RT-RPA was as sensitive as real-time RT-PCR (10 RNA molecules), rapid (3-7 minutes) and mobile (using tubescanner weighing 1kg). The MERS-CoV RT-RPA showed cross-detection neither of any of the RNAs of several coronaviruses and respiratory viruses affecting humans nor of the human genome. The developed isothermal real-time RT-RPA is ideal for rapid mobile molecular MERS-CoV monitoring in acute patients and may also facilitate the search for the animal reservoir of MERS-CoV

Eigenvalue estimates for Schroedinger operators on metric trees

We consider Schroedinger operators on regular metric trees and prove Lieb-Thirring and Cwikel-Lieb-Rozenblum inequalities for their negative eigenvalues. The validity of these inequalities depends on the volume growth of the tree. We show that the bounds are valid in the endpoint case and reflect the correct order in the weak or strong coupling limit

MenaINV dysregulates cortactin phosphorylation to promote invadopodium maturation

Invadopodia, actin-based protrusions of invasive carcinoma cells that focally activate extracellular matrix-degrading proteases, are essential for the migration and intravasation of tumor cells during dissemination from the primary tumor. We have previously shown that cortactin phosphorylation at tyrosine residues, in particular tyrosine 421, promotes actin polymerization at newly-forming invadopodia, promoting their maturation to matrix-degrading structures. However, the mechanism by which cells regulate the cortactin tyrosine phosphorylation-dephosphorylation cycle at invadopodia is unknown. Mena, an actin barbed-end capping protein antagonist, is expressed as various splice-isoforms. The MenaINV isoform is upregulated in migratory and invasive sub-populations of breast carcinoma cells, and is involved in tumor cell intravasation. Here we show that forced MenaINV expression increases invadopodium maturation to a far greater extent than equivalent expression of other Mena isoforms. MenaINV is recruited to invadopodium precursors just after their initial assembly at the plasma membrane, and promotes the phosphorylation of cortactin tyrosine 421 at invadopodia. In addition, we show that cortactin phosphorylation at tyrosine 421 is suppressed by the phosphatase PTP1B, and that PTP1B localization to the invadopodium is reduced by MenaINV expression. We conclude that MenaINV promotes invadopodium maturation by inhibiting normal dephosphorylation of cortactin at tyrosine 421 by the phosphatase PTP1B.United States. National Institutes of Health (CA150344)United States. National Institutes of Health (CA100324

The Berry-Keating operator on L^2(\rz_>, x) and on compact quantum graphs with general self-adjoint realizations

The Berry-Keating operator H_{\mathrm{BK}}:= -\ui\hbar(x\frac{ \phantom{x}}{ x}+{1/2}) [M. V. Berry and J. P. Keating, SIAM Rev. 41 (1999) 236] governing the Schr\"odinger dynamics is discussed in the Hilbert space L^2(\rz_>, x) and on compact quantum graphs. It is proved that the spectrum of $H_{\mathrm{BK}}$ defined on L^2(\rz_>, x) is purely continuous and thus this quantization of $H_{\mathrm{BK}}$ cannot yield the hypothetical Hilbert-Polya operator possessing as eigenvalues the nontrivial zeros of the Riemann zeta function. A complete classification of all self-adjoint extensions of $H_{\mathrm{BK}}$ acting on compact quantum graphs is given together with the corresponding secular equation in form of a determinant whose zeros determine the discrete spectrum of $H_{\mathrm{BK}}$. In addition, an exact trace formula and the Weyl asymptotics of the eigenvalue counting function are derived. Furthermore, we introduce the "squared" Berry-Keating operator $H_{\mathrm{BK}}^2:= -x^2\frac{ ^2\phantom{x}}{ x^2}-2x\frac{ \phantom{x}}{ x}-{1/4}$ which is a special case of the Black-Scholes operator used in financial theory of option pricing. Again, all self-adjoint extensions, the corresponding secular equation, the trace formula and the Weyl asymptotics are derived for $H_{\mathrm{BK}}^2$ on compact quantum graphs. While the spectra of both $H_{\mathrm{BK}}$ and $H_{\mathrm{BK}}^2$ on any compact quantum graph are discrete, their Weyl asymptotics demonstrate that neither $H_{\mathrm{BK}}$ nor $H_{\mathrm{BK}}^2$ can yield as eigenvalues the nontrivial Riemann zeros. Some simple examples are worked out in detail.Comment: 33p

Recombinase polymerase amplification assay for rapid detection of lumpy skin disease virus

Background  Lumpy skin disease virus (LSDV) is aCapripoxvirusinfecting cattle and Buffalos. Lumpy skin disease (LSD) leads to significant economic losses due to hide damage, reduction of milk production, mastitis, infertility and mortalities (10 %). Early detection of the virus is crucial to start appropriate outbreak control measures. Veterinarians rely on the presence of the characteristic clinical signs of LSD. Laboratory diagnostics including virus isolation, sequencing and real-time polymerase chain reaction (PCR) are performed at well-equipped laboratories. In this study, a portable, simple, and rapid recombinase polymerase amplification (RPA) assay for the detection of LSDV-genome for the use on farms was developed.  Results  The LSDV RPA assay was performed at 42 °C and detected down to 179 DNA copies/reaction in a maximum of 15 min. Unspecific amplification was observed with neither LSDV-negative samples (n= 12) nor nucleic acid preparations from orf virus, bovine papular stomatitis virus, cowpoxvirus, Peste des petits ruminants and Blue tongue virus (serotypes 1, 6 and 8). The clinical sensitivity of the LSDV RPA assay matched 100 % (n= 22) to real-time PCR results. In addition, the LSDV RPA assay detected sheep and goat poxviruses.  Conclusion  The LSDV RPA assay is a rapid and sensitive test that could be implemented in field or at quarantine stations for the identification of LSDV infected case

Development of a Flow-Trough Microarray based Reverse Transcriptase Multiplex Ligation-Dependent Probe Amplification Assay for the Detection of European Bunyaviruses

It is suspected that apart from tick-borne encephalitis virus several additional European Arboviruses such as the sandfly borne Toscana virus, sandfly fever Sicilian virus and sandfly fever Naples virus, mosquito-borne Tahyna virus, Inkoo virus, Batai virus and tick-borne Uukuniemi virus cause aseptic meningo-encephalitis or febrile disease in Europe. Currently, the microarray technology is developing rapidly and there are many efforts to apply it to infectious diseases diagnostics. In order to arrive at an assay system useful for high throughput analysis of samples from aseptic meningo-encephalitis cases the authors developed a combined multiplex ligation-dependent probe amplification and flow-through microarray assay for the detection of European Bunyaviruses. These results show that this combined assay indeed is highly sensitive, and specific for the accurate detection of multiple viruses

Development of mobile laboratory for viral hemorrhagic fever detection in Africa

Background In order to enable local response to viral haemorrhagic fever outbreaks a mobile laboratory transportable on commercial flights was developed. Methodology The development progressed from use of mobile real time RT-PCR to mobile Recombinase Polymerase Amplification (RT-RPA). The various stages of the mobile laboratory development are described. Results A brief overview of its deployments, which culminated in the first on site detection of Ebola virus disease (EVD) in March 2014 and a successful use in a campaign to roll back EVD cases in Conakry in the West-Africa Ebola virus outbreak are described. Conclusion The developed mobile laboratory successfully enabled local teams to perform rapid viral haemorrhagic fever disgnostics

Full-length genome sequence of Ntaya virus

Presentation of pyrosequencing data and phylogenetic analysis for the full genome of Ntaya virus, type virus of the Ntaya virus group of the Flaviviridae isolated in Cameroon in 1966