Using the fringe field of a clinical MRI scanner enables robotic navigation of tethered instruments in deeper vascular regions

2019 ◽  
Vol 4 (36) ◽  
pp. eaax7342 ◽  
Author(s):  
Arash Azizi ◽  
Charles C. Tremblay ◽  
Kévin Gagné ◽  
Sylvain Martel
Keyword(s):  
Insertion Site ◽  
Superconducting Magnet ◽  
Fringe Field ◽  
Friction Forces ◽  
Human Scale ◽  
Clinical Magnetic Resonance Imaging ◽  
Pulling Force ◽  
Clinical Mri

Navigating tethered instruments through the vasculatures to reach deeper physiological locations presently inaccessible would extend the applicability of many medical interventions, including but not limited to local diagnostics, imaging, and therapies. Navigation through narrower vessels requires minimizing the diameter of the instrument, resulting in a decrease of its stiffness until steerability becomes unpractical, while pushing the instrument at the insertion site to counteract the friction forces from the vessel walls caused by the bending of the instrument. To reach beyond the limit of using a pushing force alone, we report a method relying on a complementary directional pulling force at the tip created by gradients resulting from the magnetic fringe field emanating outside a clinical magnetic resonance imaging (MRI) scanner. The pulling force resulting from gradients exceeding 2 tesla per meter in a space that supports human-scale interventions allows the use of smaller magnets, such as the deformable spring as described here, at the tip of the instrument. Directional forces are achieved by robotically positioning the patient at predetermined successive locations inside the fringe field, a method that we refer to as fringe field navigation (FFN). We show through in vitro and in vivo experiments that x-ray–guided FFN could navigate microguidewires through complex vasculatures well beyond the limit of manual procedures and existing magnetic platforms. Our approach facilitated miniaturization of the instrument by replacing the torque from a relatively weak magnetic field with a configuration designed to exploit the superconducting magnet-based directional forces available in clinical MRI rooms.

scholarly journals Global Analysis of Genes Essential forFrancisella tularensisSchu S4 GrowthIn Vitroand for Fitness during Competitive Infection of Fischer 344 Rats

2019 ◽  
Vol 201 (7) ◽  
Author(s):  
Philip M. Ireland ◽  
Helen L. Bullifent ◽  
Nicola J. Senior ◽  
Stephanie J. Southern ◽  
Zheng Rong Yang ◽  
...  
Keyword(s):  
Francisella Tularensis ◽  
Insertion Site ◽  
Therapeutic Drug ◽  
Content Type ◽  
Fischer 344 ◽  
A Genome ◽  
Fischer 344 Rat ◽  
Schu S4

ABSTRACTThe highly virulent intracellular pathogenFrancisella tularensisis a Gram-negative bacterium that has a wide host range, including humans, and is the causative agent of tularemia. To identify new therapeutic drug targets and vaccine candidates and investigate the genetic basis ofFrancisellavirulence in the Fischer 344 rat, we have constructed anF. tularensisSchu S4 transposon library. This library consists of more than 300,000 unique transposon mutants and represents a transposon insertion for every 6 bp of the genome. A transposon-directed insertion site sequencing (TraDIS) approach was used to identify 453 genes essential for growthin vitro. Many of these essential genes were mapped to key metabolic pathways, including glycolysis/gluconeogenesis, peptidoglycan synthesis, fatty acid biosynthesis, and the tricarboxylic acid (TCA) cycle. Additionally, 163 genes were identified as required for fitness during colonization of the Fischer 344 rat spleen. Thisin vivoselection screen was validated through the generation of marked deletion mutants that were individually assessed within a competitive index study against the wild-typeF. tularensisSchu S4 strain.IMPORTANCEThe intracellular bacterial pathogenFrancisella tularensiscauses a disease in humans characterized by the rapid onset of nonspecific symptoms such as swollen lymph glands, fever, and headaches.F. tularensisis one of the most infectious bacteria known and following pulmonary exposure can have a mortality rate exceeding 50% if left untreated. The low infectious dose of this organism and concerns surrounding its potential as a biological weapon have heightened the need for effective and safe therapies. To expand the repertoire of targets for therapeutic development, we initiated a genome-wide analysis. This study has identified genes that are important forF. tularensisunderin vitroandin vivoconditions, providing candidates that can be evaluated for vaccine or antibacterial development.

scholarly journals Characterization of New Virulence Factors Involved in the Intracellular Growth and Survival of Burkholderia pseudomallei

2015 ◽  
Vol 84 (3) ◽  
pp. 701-710 ◽  
Author(s):  
Madeleine G. Moule ◽  
Natasha Spink ◽  
Sam Willcocks ◽  
Jiali Lim ◽  
José Afonso Guerra-Assunção ◽  
...  
Keyword(s):  
Burkholderia Pseudomallei ◽  
Insertion Site ◽  
Wild Type ◽  
Content Type ◽  
Growth And Survival ◽  
New Genes ◽  
Insertion Sites

Burkholderia pseudomallei, the causative agent of melioidosis, has complex and poorly understood extracellular and intracellular lifestyles. We used transposon-directed insertion site sequencing (TraDIS) to retrospectively analyze a transposon library that had previously been screened through a BALB/c mouse model to identify genes important for growth and survivalin vivo. This allowed us to identify the insertion sites and phenotypes of negatively selected mutants that were previously overlooked due to technical constraints. All 23 unique genes identified in the original screen were confirmed by TraDIS, and an additional 105 mutants with various degrees of attenuationin vivowere identified. Five of the newly identified genes were chosen for further characterization, and clean, unmarkedbpsl2248,tex,rpiR,bpsl1728, andbpss1528deletion mutants were constructed from the wild-type strain K96243. Each of these mutants was testedin vitroandin vivoto confirm their attenuated phenotypes and investigate the nature of the attenuation. Our results confirm that we have identified new genes important toin vivovirulence with roles in different stages ofB. pseudomalleipathogenesis, including extracellular and intracellular survival. Of particular interest, deletion of the transcription accessory protein Tex was shown to be highly attenuating, and thetexmutant was capable of providing protective immunity against challenge with wild-typeB. pseudomallei, suggesting that the genes identified in our TraDIS screen have the potential to be investigated as live vaccine candidates.

Safety of Hybrid Electrodes for Single-Neuron Recordings in Humans

Neurosurgery ◽  
2013 ◽  
Vol 73 (1) ◽  
pp. 78-85 ◽  
Author(s):  
Stefan Hefft ◽  
Armin Brandt ◽  
Stefan Zwick ◽  
Dominik von Elverfeldt ◽  
Irina Mader ◽  
...  
Keyword(s):  
Single Neuron ◽  
Electrode Implantation ◽  
Intraoperative Computed Tomography ◽  
Postoperative Mri ◽  
Depth Electrodes ◽  
Magnetic Resonance Imaging Mri ◽  
Clinical Mri

Abstract BACKGROUND: Intracranial in vivo recordings of individual neurons in humans are increasingly performed for a better understanding of the mechanisms of epileptogenesis and of the neurobiological basis of cognition. So far, information about the safety of stereotactic implantations and of magnetic resonance imaging (MRI) with hybrid depth electrodes is scarce. OBJECTIVE: The aim of this study was to assess neurosurgical safety of implantations, recordings, and imaging using hybrid electrodes in humans. METHODS: Perioperative and long-term safety of implantation of a total of 88 hybrid depth electrodes with integrated microwires was assessed retrospectively in 25 consecutive epilepsy patients who underwent implantation of electrodes from 2007 to 2011 based on electronically stored charts. Safety aspects of MRI are reported from both in vitro and in vivo investigations. Precision of electrode implantation is evaluated based on intraoperative computed tomography and pre- and postoperative MRI. RESULTS: There was no clinically relevant morbidity associated with the use of hybrid electrodes in any of the patients. Precision of recordings from the targets aimed at was similar to that of standard depth electrodes. In vitro studies demonstrated the absence of relevant heating of hybrid electrodes with newly designed connectors with MRI at 1.5 T, corresponding to well-tolerated clinical MRI in patients. CONCLUSION: Given the technical approach described here, precise targeting and safe use are possible with hybrid electrodes containing microwires for in vivo recording of human neuronal units.

scholarly journals Lentiviral vectors containing an enhancer-less ubiquitously acting chromatin opening element (UCOE) provide highly reproducible and stable transgene expression in hematopoietic cells

Blood ◽  
2007 ◽  
Vol 110 (5) ◽  
pp. 1448-1457 ◽  
Author(s):  
Fang Zhang ◽  
Susannah I. Thornhill ◽  
Steven J. Howe ◽  
Meera Ulaganathan ◽  
Axel Schambach ◽  
...  
Keyword(s):  
Transgene Expression ◽  
Lentiviral Vector ◽  
Severe Combined Immunodeficiency ◽  
Insertion Site ◽  
Housekeeping Genes ◽  
Enhancer Activity ◽  
Position Effects ◽  
Chromatin Opening

AbstractUbiquitously acting chromatin opening elements (UCOEs) consist of methylation-free CpG islands encompassing dual divergently transcribed promoters of housekeeping genes that have been shown to confer resistance to transcriptional silencing and to produce consistent and stable transgene expression in tissue culture systems. To develop improved strategies for hematopoietic cell gene therapy, we have assessed the potential of the novel human HNRPA2B1-CBX3 UCOE (A2UCOE) within the context of a self-inactivating (SIN) lentiviral vector. Unlike viral promoters, the enhancer-less A2UCOE gave rise to populations of cells that expressed a reporter transgene at a highly reproducible level. The efficiency of expression per vector genome was also markedly increased in vivo compared with vectors incorporating either spleen focus-forming virus (SFFV) or cytomegalovirus (CMV) promoters, suggesting a relative resistance to silencing. Furthermore, an A2UCOE-IL2RG vector fully restored the IL-2 signaling pathway within IL2RG-deficient human cells in vitro and successfully rescued the X-linked severe combined immunodeficiency (SCID-X1) phenotype in a mouse model of this disease. These data indicate that the A2UCOE displays highly reliable transcriptional activity within a lentiviral vector, largely overcoming insertion-site position effects and giving rise to therapeutically relevant levels of gene expression. These properties are achieved in the absence of classic enhancer activity and therefore may confer a high safety profile.

scholarly journals First Streptococcus pyogenes Signature-Tagged Mutagenesis Screen Identifies Novel Virulence Determinants

2009 ◽  
Vol 77 (5) ◽  
pp. 1854-1865 ◽  
Author(s):  
Anne E. Kizy ◽  
Melody N. Neely
Keyword(s):  
Streptococcus Pyogenes ◽  
Large Scale ◽  
Virulence Genes ◽  
Insertion Site ◽  
Disease Model ◽  
Invasive Infection ◽  
Virulence Determinants ◽  
Infectious Disease Model

ABSTRACT The virulence of bacterial pathogens is a complex process that requires the dynamic expression of many genes for the pathogens to invade and circumvent host defenses, as well as to proliferate in vivo. In this study, we employed a large-scale screen, signature-tagged mutagenesis (STM), to identify Streptococcus pyogenes virulence genes important for pathogenesis within the host. Approximately 1,200 STM mutants were created and screened using the zebrafish infectious disease model. The transposon insertion site was identified for 29 of the 150 mutants that were considered attenuated for virulence. Previously reported streptococcal virulence genes, such as mga, hasA, amrA, smeZ, and two genes in the sil locus, were identified, confirming the utility of the model for revealing genes important for virulence. Multiple genes not previously implicated in virulence were also identified, including genes encoding putative transporters, hypothetical cytosolic proteins, and macrolide efflux pumps. The STM mutant strains display various levels of attenuation, and multiple separate insertions were identified in either the same gene or the same locus, suggesting that these factors are important for this type of acute, invasive infection. We further examined two such genes, silB and silC of a putative quorum-sensing regulon, and determined that they are significant virulence factors in our model of necrotizing fasciitis. sil locus promoter expression was examined under various in vitro conditions, as well as in zebrafish tissues, and was found to be differentially induced. This study was a unique investigation of S. pyogenes factors required for successful invasive infection.

scholarly journals An NMR relaxometry approach for quantitative investigation of the transchelation of gadolinium ions from GBCAs to a competing macromolecular chelator

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Patrick Werner ◽  
Matthias Taupitz ◽  
Leif Schröder ◽  
Patrick Schuenke
Keyword(s):  
Time Resolved ◽  
Nmr Relaxometry ◽  
Clinical Magnetic Resonance Imaging ◽  
Heparin Complexes ◽  
Magnetic Resonance Imaging Mri ◽  
Indirect Impacts ◽  
Underlying Processes

AbstractGadolinium-based contrast agents (GBCAs) have been used in clinical Magnetic Resonance Imaging (MRI) for more than 30 years. However, there is increasing evidence that their dissociation in vivo leads to long-term depositions of gadolinium ions in the human body. In vitro experiments provide critical insights into kinetics and thermodynamic equilibria of underlying processes, which give hints towards the in vivo situation. We developed a time-resolved MRI relaxometry-based approach that exploits distinct relaxivities of Gd3+ in different molecular environments. Its applicability to quantify the transmetallation of GBCAs, the binding of Gd3+ to competing chelators, and the combined transchelation process is demonstrated. Exemplarily, the approach is applied to investigate two representative GBCAs in the presence of Zn2+ and heparin, which is used as a model for a macromolecular and physiologically occurring chelator. Opposing indirect impacts of heparin on increasing the kinetic stability but reducing the thermodynamic stability of GBCAs are observed. The relaxivity of resulting Gd-heparin complexes is shown to be essentially increased compared to that of the parent GBCAs so that they might be one explanation for observed long-term MRI signal enhancement in vivo. In forthcoming studies, the presented method could help to identify the most potent Gd-complexing macromolecular species.

scholarly journals Vision-Based Suture Tensile Force Estimation in Robotic Surgery

Sensors ◽  
2020 ◽  
Vol 21 (1) ◽  
pp. 110
Author(s):  
Won-Jo Jung ◽  
Kyung-Soo Kwak ◽  
Soo-Chul Lim
Keyword(s):  
Force Feedback ◽  
Short Term Memory ◽  
Tensile Force ◽  
Force Sensor ◽  
Interaction Force ◽  
Force Estimation ◽  
Training Experience ◽  
Pulling Force

Compared to laparoscopy, robotics-assisted minimally invasive surgery has the problem of an absence of force feedback, which is important to prevent a breakage of the suture. To overcome this problem, surgeons infer the suture force from their proprioception and 2D image by comparing them to the training experience. Based on this idea, a deep-learning-based method using a single image and robot position to estimate the tensile force of the sutures without a force sensor is proposed. A neural network structure with a modified Inception Resnet-V2 and Long Short Term Memory (LSTM) networks is used to estimate the suture pulling force. The feasibility of proposed network is verified using the generated DB, recording the interaction under the condition of two different artificial skins and two different situations (in vivo and in vitro) at 13 viewing angles of the images by changing the tool positions collected from the master-slave robotic system. From the evaluation conducted to show the feasibility of the interaction force estimation, the proposed learning models successfully estimated the tensile force at 10 unseen viewing angles during training.

scholarly journals In Vitro Reconstitution of Dynein Force Exertion in Bulk Cytoplasm

2020 ◽  
Author(s):  
Héliciane Palenzuela ◽  
Benjamin Lacroix ◽  
Jérémy Sallé ◽  
Katsuhiko Minami ◽  
Tomohiro Shima ◽  
...  
Keyword(s):  
Viscous Medium ◽  
Hydrodynamic Interactions ◽  
Cell Cortex ◽  
Cellular Processes ◽  
In Vitro Reconstitution ◽  
Reconstituted System ◽  
Pulling Force ◽  
Glass Surfaces

SUMMARYThe forces generated by Microtubules (MTs) and their associated motors orchestrate essential cellular processes ranging from vesicular trafficking to centrosome positioning [1, 2]. To date, most studies have focused on force exertion from motors anchored on a static surface, such as the cell cortex in vivo or glass surfaces in vitro [2–4]. However, motors also transport large cargos and endomembrane networks, whose hydrodynamic interactions with the viscous cytoplasm should generate sizable forces in bulk. Such forces may contribute to MT aster centration, organization and orientation [5–14], but have yet to be evidenced and studied in a minimal reconstituted system. By developing a bulk motility assay, based on stabilized MTs and dynein-coated beads freely floating in a viscous medium away from any surface, we demonstrate that the motion of a cargo exerts a pulling force on the MT and propels it in opposite direction. Quantification of resulting MT movements for different motors, motor velocities, over a range of cargo size and medium viscosities, shows that the efficiency of this mechanism is primarily determined by cargo size and MT length. Forces exerted by cargos are additive, allowing us to recapitulate tug-of-war situations, or bi-dimensional motions of minimal asters. These data also reveal unappreciated effects of the nature of viscous crowders and hydrodynamic interactions between cargos and MTs, likely relevant to understand this mode of force exertion in living cells. This study places endomembrane transport as a significant mode of MT force exertion with far-reaching consequences for cellular organization.

scholarly journals Protein Translation Inhibition is Involved in the Activity of the Pan-PIM Kinase Inhibitor PIM447 in Combination with Pomalidomide-Dexamethasone in Multiple Myeloma

Cancers ◽  
2020 ◽  
Vol 12 (10) ◽  
pp. 2743
Author(s):  
Teresa Paíno ◽  
Lorena González-Méndez ◽  
Laura San-Segundo ◽  
Luis A. Corchete ◽  
Susana Hernández-García ◽  
...  
Keyword(s):  
Multiple Myeloma ◽  
Kinase Inhibitor ◽  
Leukemia Virus ◽  
Insertion Site ◽  
Protein Translation ◽  
Myeloma Cell ◽  
In Vivo Studies ◽  
Triple Combination

Background: Proviral Insertion site for Moloney murine leukemia virus (PIM) kinases are overexpressed in hematologic malignancies, including multiple myeloma. Previous preclinical data from our group demonstrated the anti-myeloma effect of the pan-PIM kinase inhibitor PIM447. Methods: Based on those data, we evaluate here, by in vitro and in vivo studies, the activity of the triple combination of PIM447 + pomalidomide + dexamethasone (PIM-Pd) in multiple myeloma. Results: Our results show that the PIM-Pd combination exerts a potent anti-myeloma effect in vitro and in vivo, where it markedly delays tumor growth and prolongs survival of treated mice. Mechanism of action studies performed in vitro and on mice tumor samples suggest that the combination PIM-Pd inhibits protein translation processes through the convergent inhibition of c-Myc and mTORC1, which subsequently disrupts the function of eIF4E. Interestingly the MM pro-survival factor IRF4 is also downregulated after PIM-Pd treatment. As a whole, all these molecular changes would promote cell cycle arrest and deregulation of metabolic pathways, including glycolysis and lipid biosynthesis, leading to inhibition of myeloma cell proliferation. Conclusions: Altogether, our data support the clinical evaluation of the triple combination PIM-Pd for the treatment of patients with multiple myeloma.

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