Rare but diverse off-target and somatic mutations found in field and greenhouse grown trees expressing CRISPR/Cas9.

CRISPR gene editing, while highly efficient in creating desired mutations, also has the potential to cause off-target mutations. This risk is especially high in clonally propagated plants, where editing reagents may remain in the genome for long periods of time or in perpetuity. We studied a diverse population of and trees that had CRISPR/Cas9-containing transgenes that targeted one or two types of floral development genes, homologs of and .

A phase Ib trial of pembrolizumab plus paclitaxel or flat-dose capecitabine in 1st/2nd line metastatic triple-negative breast cancer.

Chemoimmunotherapy with anti-programmed cell death 1/ligand 1 and cytotoxic chemotherapy is a promising therapeutic modality for women with triple-negative breast cancer, but questions remain regarding optimal chemotherapy backbone and biomarkers for patient selection. We report final outcomes from a phase Ib trial evaluating pembrolizumab (200 mg IV every 3 weeks) with either weekly paclitaxel (80 mg/m weekly) or flat-dose capecitabine (2000 mg orally twice daily for 7 days of every 14-day cycle) in the 1st/2nd line setting. The primary endpoint is safety (receipt of 2 cycles without grade III/IV toxicities requiring discontinuation or ≥21-day delays).

Paediatric adverse event rating scale: a measure of safety or efficacy? Novel analysis from the MADDY study.

Objective: The Pediatric Adverse Event Rating Scale (PAERS) measured adverse events of children aged 6-12 years with ADHD and emotional dysregulation in the Micronutrients for ADHD in Youth (MADDY) study, an eight week multi-site randomized clinical trial of a broad-spectrum multinutrient treatment. Treatment sensitivity of the PAERS was assessed by calculating the treatment difference in change of the item scores from baseline to end of the RCT.

Methods: Principal component analysis retained 14 "adverse events" (out of 43 in the PAERS) that reflected ADHD symptoms and emotional dysregulation and was used to group the variables of interest.

Changes in T-cell subsets and clonal repertoire during chemoimmunotherapy with pembrolizumab and paclitaxel or capecitabine for metastatic triple-negative breast cancer.

Background: Chemoimmunotherapy is a standard treatment for triple-negative breast cancer (TNBC), however, the impacts of different chemotherapies on T-cell populations, which could correlate with clinical activity, are not known. Quantifying T-cell populations with flow cytometry and T-cell receptor (TCR) immunosequencing may improve our understanding of how chemoimmunotherapy affects T-cell subsets, and to what extent clonal shifts occur during treatment. TCR immunosequencing of intratumoral T cells may facilitate the identification and monitoring of putatively tumor-reactive T-cell clones within the blood.

A Phase Ib Study of Preoperative, Locoregional IRX-2 Cytokine Immunotherapy to Prime Immune Responses in Patients with Early-Stage Breast Cancer.

Purpose: To evaluate the safety and feasibility of preoperative locoregional cytokine therapy (IRX-2 regimen) in early-stage breast cancer, and to evaluate for intratumoral and peripheral immunomodulatory activity.

Patients And Methods: Sixteen patients with stage I-III early-stage breast cancer (any histology type) indicated for surgical lumpectomy or mastectomy were enrolled to receive preoperative locoregional immunotherapy with the IRX-2 cytokine biological (2 mL subcutaneous × 10 days to periareolar skin). The regimen also included single-dose cyclophosphamide (300 mg/m) on day 1 to deplete T-regulatory cells and oral indomethacin to modulate suppressive myeloid subpopulations.

Efficacy and Safety of Honey Bee Venom ( Dermal Injections to Treat Osteoarthritis Knee Pain and Physical Disability: A Randomized Controlled Trial.

To evaluate purified honey bee () venom (HBV) biotherapy for the treatment of osteoarthritis (OA) knee pain and physical function. Five hundred and thirty-eight patients with Kellgren/Lawrence grade 1-3 radiographic knee OA and Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC) pain score ≥2 were randomized 1:2 to either control ("histamine") or HBV in this double-blind study. After a dose escalation period, patients received 12 weekly dermal injections of control ("histamine") or HBV.

Implementation and Validation of an Automated Flow Cytometry Analysis Pipeline for Human Immune Profiling.

Automated reagent preparation, sample processing, and data acquisition have increased the rate at which flow cytometry data can be generated. Furthermore, advances in technology and flow cytometry instrumentation continually increase the complexity and dimensionality of this data. Together, this leads to increased pressure on manual data analysis, which has inherent limitations including subjectivity of the analyst and the length of time needed for data processing.

Invisible Flashes Alter Perceived Sound Location.

Information integration across the senses is fundamental for effective interactions with our environment. The extent to which signals from different senses can interact in the absence of awareness is controversial. Combining the spatial ventriloquist illusion and dynamic continuous flash suppression (dCFS), we investigated in a series of two experiments whether visual signals that observers do not consciously perceive can influence spatial perception of sounds.

A spatially collocated sound thrusts a flash into awareness.

To interact effectively with the environment the brain integrates signals from multiple senses. It is currently unclear to what extent spatial information can be integrated across different senses in the absence of awareness. Combining dynamic continuous flash suppression (CFS) and spatial audiovisual stimulation, the current study investigated whether a sound facilitates a concurrent visual flash to elude flash suppression and enter perceptual awareness depending on audiovisual spatial congruency.

CpG preconditioning regulates miRNA expression that modulates genomic reprogramming associated with neuroprotection against ischemic injury.

Cytosine-phosphate-guanine (CpG) preconditioning reprograms the genomic response to stroke to protect the brain against ischemic injury. The mechanisms underlying genomic reprogramming are incompletely understood. MicroRNAs (miRNAs) regulate gene expression; however, their role in modulating gene responses produced by CpG preconditioning is unknown.

Naturalistic stimulus structure determines the integration of audiovisual looming signals in binocular rivalry.

Rapid integration of biologically relevant information is crucial for the survival of an organism. Most prominently, humans should be biased to attend and respond to looming stimuli that signal approaching danger (e.g.

Poly-ICLC preconditioning protects the blood-brain barrier against ischemic injury in vitro through type I interferon signaling.

Preconditioning with a low dose of harmful stimulus prior to injury induces tolerance to a subsequent ischemic challenge resulting in neuroprotection against stroke. Experimental models of preconditioning primarily focus on neurons as the cellular target of cerebral protection, while less attention has been paid to the cerebrovascular compartment, whose role in the pathogenesis of ischemic brain injury is crucial. We have shown that preconditioning with polyinosinic polycytidylic acid (poly-ICLC) protects against cerebral ischemic damage.

Interactions between apparent motion rivalry in vision and touch.

In multistable perception, the brain alternates between several perceptual explanations of ambiguous sensory signals. It is unknown whether multistable processes can interact across the senses. In the study reported here, we presented subjects with unisensory (visual or tactile), spatially congruent visuotactile, and spatially incongruent visuotactile apparent motion quartets.

Toll-like receptor 7 preconditioning induces robust neuroprotection against stroke by a novel type I interferon-mediated mechanism.

Background And Purpose: Systemic administration of Toll-like receptor (TLR) 4 and TLR9 agonists before cerebral ischemia have been shown to reduce ischemic injury by reprogramming the response of the brain to stroke. Our goal was to explore the mechanism of TLR-induced neuroprotection by determining whether a TLR7 agonist also protects against stroke injury.

Methods: C57Bl/6, TNF(-/-), interferon (IFN) regulatory factor 7(-/-), or type I IFN receptor (IFNAR)(-/-) mice were subcutaneously administered the TLR7 agonist Gardiquimod (GDQ) 72 hours before middle cerebral artery occlusion.

Audiovisual interactions in binocular rivalry.

When the two eyes are presented with dissimilar images, human observers report alternating percepts-a phenomenon coined binocular rivalry. These perceptual fluctuations reflect competition between the two visual inputs both at monocular and binocular processing stages. Here we investigated the influence of auditory stimulation on the temporal dynamics of binocular rivalry.

Coherent tunneling adiabatic passage with the alternating coupling scheme.

The use of adiabatic passage techniques to mediate particle transport through real space, rather than phase space, is becoming an interesting possibility. We have investigated the properties of coherent tunneling adiabatic passage (CTAP) with alternating tunneling matrix elements. This coupling scheme, not previously considered in the donor in silicon paradigm, provides an interesting route to long-range quantum transport.

Signal transduction and procoagulant state of human cord blood--progenitor-derived endothelial cells after interleukin-1alpha stimulation.

Isolation of endothelial progenitors from human umbilical cord blood generated great hope in vascular tissue engineering. However, before clinical use, progenitor derived endothelial cells (PDECs) have to be compared with mature endothelial cells (ECs). The aim of this study was to explore the behavior of PDECs exposed to a proinflammatory cytokine (interleukin-1alpha; IL-1alpha) according to the mitogen-activated protein (MAP) kinase and nuclear factor (NF)-kappaB signal transduction pathways as well as procoagulant activity (PCA).

The Steubenville Comprehensive Air Monitoring Program (SCAMP): concentrations and solubilities of PM(2.5) trace elements and their implications for source apportionment and health research.

The elemental compositions of the water-soluble and acid-digestible fractions of 24-hr integrated fine particulate matter (PM(2.5)) samples collected in Steubenville, OH, from 2000 to 2002 were determined using dynamic reaction cell inductively coupled plasma-mass spectrometry. The water-soluble elemental compositions of PM(2.

3D shape perception from combined depth cues in human visual cortex.

Our perception of the world's three-dimensional (3D) structure is critical for object recognition, navigation and planning actions. To accomplish this, the brain combines different types of visual information about depth structure, but at present, the neural architecture mediating this combination remains largely unknown. Here, we report neuroimaging correlates of human 3D shape perception from the combination of two depth cues.

Ex vivo expansion of haematopoietic progenitors on an endothelialized hydroxyapatite matrix.

Autologous haematopoietic progenitor cell (HPC) transplantation is increasingly used to restore haematopoiesis after high-dose chemotherapy treatments. The present study was designed to analyse the ability of hydroxyapatite (HAP) seeded with endothelial cells (EC) to support the proliferation and differentiation of CD34+ HPC in static culture conditions. HAP is endothelializable as assessed by scanning electron microscopy and time-course DNA synthesis analysis using tritiated thymidine incorporated in EC isolated from human umbilical vein cord.

Human primary endothelial cells stimulate human osteoprogenitor cell differentiation.

Bone development and remodeling depend on complex interactions between bone-forming osteoblasts and other cells present within the bone microenvironment, particularly endothelial cells that may be pivotal members of a complex interactive communication network in bone. While cell cooperation was previously established between Human OsteoProgenitor cells (HOP) and Human Umbilical Vein Endothelial Cells (HUVEC) the aim of our study was to investigate if this interaction is specific to Human Endothelial cell types (ECs) from different sources. Osteoblastic cell differentiation analysis performed using different co-culture models with direct contact revealed that Alkaline Phosphatase (Al-P) activity was only increased by the direct contact of HOP with human primary vascular endothelial cell types including endothelial precursor cells (EPCs) isolated from blood cord, endothelial cells from Human Saphen Vein (HSV) while a transformed cell line, the Human Bone Marrow Endothelial Cell Line (HBMEC) did not modify osteoblastic differentiation of HOP.

Human bone marrow endothelial cells: a new identified source of B-type natriuretic peptide.

B-type natriuretic peptide (BNP) is a hormone mainly secreted by cardiac ventricle myocytes and which is increased in cardiac diseases. Moreover, BNP expression has been shown in various cell/tissue types. Six different human endothelial cell (EC) culture models arising from macro and microcirculation either primary cultures or cell lines were cultured and screened for BNP presence and secretion.

Evaluation of an in vitro endothelialized vascular graft under pulsatile shear stress with a novel radiolabeling procedure.

Objective: To improve the hemocompatibility of vascular grafts, endothelial cell (EC) seeding of biomaterials prior to implantation is critical. The current in vitro study was designed to investigate such a feasibility on a collagen-coated heparin-bonded graft and to evaluate cell detachment upon pulsatile shear stress.

Materials And Methods: Endothelial cells (EA-hy-926) were seeded onto grafts.

Expansion and differentiation of haemopoietic progenitor cells on endothelialized hydroxyapatite under static conditions.

We have analysed the potential of an endothelialized hydroxyapatite matrix (HAP), a synthetic bone substitute, as a cellularized support for the expansion of haemopoietic progenitor cells. Scanning electron microscopy (SEM) showed that the endothelial cells (EC) tended to form a monolayer fitting closely to the matrix, and that the progenitors adhered to the EC layer. Endothelialized HAP supported the proliferation and differentiation of the progenitors with the addition of the exogenous cytokines IL-1, and IL-3.

Ability of various inserts to promote endothelium cell culture for the establishment of coculture models.

To select an insert suitable for human umbilical vein endothelial cell (HUVEC) culture, we compared several available inserts of 0.2 to 0.45 micron porosity: Cellagen (ICN), Transwell-COL (Costar), Millicell-HA and CM (Millipore), Anopore (Nunc), Cyclopore (Falcon) in comparison with a control surface (Thermanox).