34,202 research outputs found
Optical techniques for 3D surface reconstruction in computer-assisted laparoscopic surgery
One of the main challenges for computer-assisted surgery (CAS) is to determine the intra-opera- tive morphology and motion of soft-tissues. This information is prerequisite to the registration of multi-modal patient-specific data for enhancing the surgeon’s navigation capabilites by observ- ing beyond exposed tissue surfaces and for providing intelligent control of robotic-assisted in- struments. In minimally invasive surgery (MIS), optical techniques are an increasingly attractive approach for in vivo 3D reconstruction of the soft-tissue surface geometry. This paper reviews the state-of-the-art methods for optical intra-operative 3D reconstruction in laparoscopic surgery and discusses the technical challenges and future perspectives towards clinical translation. With the recent paradigm shift of surgical practice towards MIS and new developments in 3D opti- cal imaging, this is a timely discussion about technologies that could facilitate complex CAS procedures in dynamic and deformable anatomical regions
Pattern Matching and Discourse Processing in Information Extraction from Japanese Text
Information extraction is the task of automatically picking up information of
interest from an unconstrained text. Information of interest is usually
extracted in two steps. First, sentence level processing locates relevant
pieces of information scattered throughout the text; second, discourse
processing merges coreferential information to generate the output. In the
first step, pieces of information are locally identified without recognizing
any relationships among them. A key word search or simple pattern search can
achieve this purpose. The second step requires deeper knowledge in order to
understand relationships among separately identified pieces of information.
Previous information extraction systems focused on the first step, partly
because they were not required to link up each piece of information with other
pieces. To link the extracted pieces of information and map them onto a
structured output format, complex discourse processing is essential. This paper
reports on a Japanese information extraction system that merges information
using a pattern matcher and discourse processor. Evaluation results show a high
level of system performance which approaches human performance.Comment: See http://www.jair.org/ for any accompanying file
Production and analysis of synthetic Cascade variants
CRISPR (clustered regularly interspaced short palindromic repeats)-Cas (CRISPR assoziiert) ist ein
adaptives Immunsystem in Archaeen und Bakterien, das fremdes genetisches Material mit Hilfe von
Ribonukleoprotein-Komplexen erkennt und zerstört. Diese Komplexe bestehen aus einer CRISPR RNA
(crRNA) und Cas Proteinen. CRISPR-Cas Systeme sind in zwei Hauptklassen und mehrere Typen
unterteilt, abhängig von den beteiligten Cas Proteinen. In Typ I Systemen sucht ein Komplex namens
Cascade (CRISPR associated complex for antiviral defence) nach eingedrungener viraler DNA während
einer Folgeinfektion und bindet die zu der eingebauten crRNA komplementäre Sequenz. Anschließend
wird die Nuklease/Helikase Cas3 rekrutiert, welche die virale DNA degradiert (Interferenz).
Das Typ I System wird in mehrere Subtypen unterteilt, die Unterschiede im Aufbau von Cascade
vorweisen. Im Fokus dieser Arbeit steht eine minimale Cascade-Variante aus Shewanella putrefaciens
CN-32. Im Vergleich zur gut untersuchten Typ I-E Cascade aus Escherichia coli fehlen in diesem Komplex
zwei Untereinheiten, die gewöhnlicher Weise für die Zielerkennung benötigt werden. Dennoch ist der
Komplex aktiv. Rekombinante I-Fv Cascade wurde bereits aus E. coli aufgereinigt und es war möglich,
den Komplex zu modifizieren, indem das Rückgrat entweder verlängert oder verkürzt wurde. Dadurch
wurden synthetische Varianten mit veränderter Protein-Stöchiometrie erzeugt.
In der vorliegenden Arbeit wurde I-Fv Cascade weiter mit in vitro Methoden untersucht. So wurde die
Bindung von Ziel-DNA beobachtet und die 3D Struktur zeigt, dass strukturelle Veränderungen im
Komplex die fehlenden Untereinheiten ersetzen, möglicherweise um viralen Anti-CRISPR Proteinen zu
entgehen. Die Nuklease/Helikase dieses Systems, Cas2/3fv, ist eine Fusion des Cas3 Proteins mit dem
Interferenz-unabhängigen Protein Cas2. Ein unabhängiges Cas3fv ohne Cas2 Untereinheit wurde
aufgereinigt und in vitro Assays zeigten, dass dieses Protein sowohl freie ssDNA als auch Cascadegebundene Substrate degradiert. Das komplette Cas2/3fv Protein bildet einen Komplex mit dem Protein
Cas1 und zeigt eine reduzierte Aktivität gegenüber freier ssDNA, möglicherweise als
Regulationsmechanismus zur Vermeidung von unspezifischer Aktivität.
Weiterhin wurde ein Prozess namens „RNA wrapping“ etabliert. Synthetische Cascade-Komplexe
wurden erzeugt, in denen die grundlegende RNA-Bindung des charakteristischen Cas7fv RückgratProteins auf eine ausgewählte RNA gelenkt wird. Diese spezifische Komplexbildung kann in vivo durch
eine Repeat-Sequenz der crRNA stromaufwärts der Zielsequenz und durch Bindung des Cas5fv Proteins
initiiert werden. Die erzeugten Komplexe beinhalten die ersten 100 nt der markierten RNA, die
anschließend isoliert werden kann. Innerhalb der Komplexe ist die RNA stabilisiert und geschützt vor
Degradation durch RNasen. Komplexbildung kann außerdem genutzt werden, um ReportergenTranskripte stillzulegen. Zusätzlich wurden erste Hinweise geliefert, dass das Rückgrat der synthetischen
Komplexe durch Fusion mit weiteren Reporterproteinen modifiziert werden kann.CRISPR (clustered regularly interspaced short palindromic repeats)-Cas (CRISPR associated) is an
adaptive immune system of Archaea and Bacteria. It is able to target and destroy foreign genetic
material with ribonucleoprotein complexes consisting of CRISPR RNAs (crRNAs) and certain Cas proteins.
CRISPR-Cas systems are classified in two major classes and multiple types, according to the involved Cas
proteins. In type I systems, a ribonucleoprotein complex called Cascade (CRISPR associated complex for
antiviral defence) scans for invading viral DNA during a recurring infection and binds the sequence
complementary to the incorporated crRNA. After target recognition, the nuclease/helicase Cas3 is
recruited and subsequently destroys the viral DNA in a step termed interfere nce.
Multiple subtypes of type I exist that show differences in the Cascade composition. This work focuses on
a minimal Cascade variant found in Shewanella putrefaciens CN-32. In comparison to the well-studied
type I-E Cascade from Escherichia coli, this complex is missing two proteins usually required for target
recognition, yet it is still able to provide immunity. Recombinant I-Fv Cascade was previously purified
from E. coli and it was possible to modulate the complex by extending or shortening the backbone,
resulting in synthetic variants with altered protein stoichiometry.
In the present study, I-Fv Cascade was further analyzed by in vitro methods. Target binding was
observed and the 3D structure revealed structural variations that replace the missing subunits,
potentially to evade viral anti-CRISPR proteins. The nuclease/helicase of this system, Cas2/3fv, is a fusion
of the Cas3 protein with the interference-unrelated protein Cas2. A standalone Cas3fv was purified
without the Cas2 domain and in vitro cleavage assays showed that Cas3fv degrades both free ssDNA as
well as Cascade-bound substrates. The complete Cas2/3fv protein forms a complex with the protein
Cas1 and was shown to reduce cleave of free ssDNA, potentially as a regulatory mechanism against
unspecific cleavage.
Furthermore, we established a process termed “RNA wrapping”. Synthetic Cascade assemblies can be
created by directing the general RNA-binding ability of the characteristic Cas7fv backbone protein on an
RNA of choice such as reporter gene transcripts. Specific complex formation can be initiated in vivo by
including a repeat sequence from the crRNA upstream a given target sequence and binding of the
Cas5fv protein. The created complexes contain the initial 100 nt of the tagged RNA which can be
isolated afterwards. While incorporated in complexes, RNA is stabilized and protected from degradation
by RNases. Complex formation can be used to silence reporter gene transcripts. Furthermore, we
provided initial indications that the backbone of synthetic complexes can be modified by addition of
reporter proteins
Robust visualization and discrimination of nanoparticles by interferometric imaging
Single-molecule and single-nanoparticle biosensors are a growing frontier in diagnostics. Digital biosensors are those which enumerate all specifically immobilized biomolecules or biological nanoparticles, and thereby achieve limits of detection usually beyond the reach of ensemble measurements. Here we review modern optical techniques for single nanoparticle detection and describe the single-particle interferometric reflectance imaging sensor (SP-IRIS). We present challenges associated with reliably detecting faint nanoparticles with SP-IRIS, and describe image acquisition processes and software modifications to address them. Specifically, we describe a image acquisition processing method for the discrimination and accurate counting of nanoparticles that greatly reduces both the number of false positives and false negatives. These engineering improvements are critical steps in the translation of SP-IRIS towards applications in medical diagnostics.R01 AI096159 - NIAID NIH HHSFirst author draf
A planning approach to the automated synthesis of template-based process models
The design-time specification of flexible processes can be time-consuming and error-prone, due to the high number of tasks involved and their context-dependent nature. Such processes frequently suffer from potential interference among their constituents, since resources are usually shared by the process participants and it is difficult to foresee all the potential tasks interactions in advance. Concurrent tasks may not be independent from each other (e.g., they could operate on the same data at the same time), resulting in incorrect outcomes. To tackle these issues, we propose an approach for the automated synthesis of a library of template-based process models that achieve goals in dynamic and partially specified environments. The approach is based on a declarative problem definition and partial-order planning algorithms for template generation. The resulting templates guarantee sound concurrency in the execution of their activities and are reusable in a variety of partially specified contextual environments. As running example, a disaster response scenario is given. The approach is backed by a formal model and has been tested in experiment
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