Viola kauaensis var.hosakae (Violaceae), a new variety of endemic Hawaiian violet

The Hawaiian endemic Viola kauaensis A. Gray has a broad distribution in bogs of Kaua`i and a limited distribution on mesic ridges in the Ko`olau Mountains of O`ahu. Based on differences in scale, the O`ahu populations of V. kauaensis had previously been described as a distinct taxon. The taxonomic status of the O`ahu populations was reevaluated through a morphometric analysis of all varieties of V. kauaensis and the morphologically similar V. vanroyenii. Morphological features of historic and freshly collected specimens of all varieties of V. kauaensis were analyzed with a principal components analysis. Populations from O`ahu represent a distinct cluster that slightly overlaps with V. kauaensis var. kauaensis. Lamina width, apex angle, and base angles contribute to the separation of the O`ahu populations from other varieties of V. kauaensis. Due to differences in scale, the O`ahu populations are described as Viola kauaensis var. hosakae, a new critically endangered taxon

A Novel High-Throughput, High-Content Three-Dimensional Assay for Determination of Tumor Invasion and Dormancy

Metastasis accounts for most cancer deaths, while dormancy of tumor cells leads to unexpected cancer recurrence. These two aspects of cancer remain relatively untreatable in part because current two-dimensional (2D) methods of high-throughput drug screening cannot quantify outcomes related to these phenotypes. Three-dimensional (3D) in-vitro tumor models are a promising alternative because they better recreate the tumor microenvironment and relevant phenotypes. However, outcome measures for high-throughput screening of these systems are often limited to single measures such as metabolic activity using assays that are not standardized or optimized for 3D models. To address this gap, the objective of this work to develop an image-based assay to measure tumor cell health, proliferation, invasion, and dormancy for high-throughput drug screening. Drug dosing experiments were performed using a novel 3D pancreatic tumor metastasis model, followed by application of various dye combinations to assess viability, proliferation, metabolic activity, and invasion. We successfully demonstrated that high throughput imaging and analysis can be performed to quantify proliferation, metabolic activity, and invasion in a single multi-output assay. Proof-of-concept experiments also revealed that while gemcitabine does effectively inhibit cancer cell proliferation it does not kill all cells and may contribute to tumor dormancy. Overall, this work using a novel 3D tumor metastasis model coupled with a multi-output assay serves as a first step toward a drug screening platform that will enable researchers to better correlate in-vitro model results with clinical outcomes related to metastasis and dormancy

A method for measuring the Neel relaxation time in a frozen ferrofluid

We report a novel method of determining the average Neel relaxation time and its temperature dependence by calculating derivatives of the measured time dependence of temperature for a frozen ferrofluid exposed to an alternating magnetic field. The ferrofluid, composed of dextran-coated Fe3O4 nanoparticles (diameter 13.7 nm +/- 4.7 nm), was synthesized via wet chemical precipitation and characterized by x-ray diffraction and transmission electron microscopy. An alternating magnetic field of constant amplitude (H0 = 20 kA/m) driven at frequencies of 171 kHz, 232 kHz and 343 kHz was used to determine the temperature dependent magnetic energy absorption rate in the temperature range from 160 K to 210 K. We found that the specific absorption rate of the ferrofluid decreased monotonically with temperature over this range at the given frequencies. From these measured data, we determined the temperature dependence of the Neel relaxation time and estimate a room-temperature magnetocrystalline anisotropy constant of 40 kJ/m3, in agreement with previously published results

Bio-Inspired Load Balancing In Large-Scale WSNs Using Pheromone Signalling

Wireless sensor networks (WSNs) consist of multiple, distributed nodes each with limited resources. With their strict resource constraints and application-specific characteristics, WSNs contain many challenging tradeoffs. This paper proposes a bioinspired load balancing approach, based on pheromone signalling mechanisms, to solve the tradeoff between service availability and energy consumption. We explore the performance consequences of the pheromone-based load balancing approach using (1) a system-level simulator, (2) deployment of real sensor testbeds to provide a competitive analysis of these evaluation methodologies. The effectiveness of the proposed algorithm is evaluated with different scenario parameters and the required performance evaluation techniques are investigated on case studies based on sound sensors

Hit and run versus long-term activation of PARP-1 by its different domains fine-tunes nuclear processes.

Poly(ADP-ribose) polymerase 1 (PARP-1) is a multidomain multifunctional nuclear enzyme involved in the regulation of the chromatin structure and transcription. PARP-1 consists of three functional domains: the N-terminal DNA-binding domain (DBD) containing three zinc fingers, the automodification domain (A), and the C-terminal domain, which includes the protein interacting WGR domain (W) and the catalytic (Cat) subdomain responsible for the poly(ADP ribosyl)ating reaction. The mechanisms coordinating the functions of these domains and determining the positioning of PARP-1 in chromatin remain unknown. Using multiple deletional isoforms of PARP-1, lacking one or another of its three domains, as well as consisting of only one of those domains, we demonstrate that different functions of PARP-1 are coordinated by interactions among these domains and their targets. Interaction between the DBD and damaged DNA leads to a short-term binding and activation of PARP-1. This hit and run activation of PARP-1 initiates the DNA repair pathway at a specific point. The long-term chromatin loosening required to sustain transcription takes place when the C-terminal domain of PARP-1 binds to chromatin by interacting with histone H4 in the nucleosome. This long-term activation of PARP-1 results in a continuous accumulation of pADPr, which maintains chromatin in the loosened state around a certain locus so that the transcription machinery has continuous access to DNA. Cooperation between the DBD and C-terminal domain occurs in response to heat shock (HS), allowing PARP-1 to scan chromatin for specific binding sites

Levels and sources of volatile organic compounds in homes of children with asthma

Many volatile organic compounds ( VOC s) are classified as known or possible carcinogens, irritants, and toxicants, and VOC exposure has been associated with the onset and exacerbation of asthma. This study characterizes VOC levels in 126 homes of children with asthma in Detroit, Michigan, USA. The total target VOC concentration ranged from 14 to 2274  μ g/m 3 (mean = 150  μ g/m 3 ; median = 91  μ g/m 3 ); 56 VOC s were quantified; and d ‐limonene, toluene, p , m ‐xylene, and ethyl acetate had the highest concentrations. Based on the potential for adverse health effects, priority VOC s included naphthalene, benzene, 1,4‐dichlorobenzene, isopropylbenzene, ethylbenzene, styrene, chloroform, 1,2‐dichloroethane, tetrachloroethene, and trichloroethylene. Concentrations varied mostly due to between‐residence and seasonal variation. Identified emission sources included cigarette smoking, solvent‐related emissions, renovations, household products, and pesticides. The effect of nearby traffic on indoor VOC levels was not distinguished. While concentrations in the Detroit homes were lower than levels found in other North American studies, many homes had elevated VOC levels, including compounds that are known health hazards. Thus, the identification and control of VOC sources are important and prudent, especially for vulnerable individuals. Actions and policies to reduce VOC exposures, for example, sales restrictions, improved product labeling, and consumer education, are recommended.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/108002/1/ina12086.pd

Human platelet lysate improves human cord blood derived ECFC survival and vasculogenesis in three dimensional (3D) collagen matrices

Human cord blood (CB) is enriched in circulating endothelial colony forming cells (ECFCs) that display high proliferative potential and in vivo vessel forming ability. Since diminished ECFC survival is known to dampen the vasculogenic response in vivo, we tested how long implanted ECFC survive and generate vessels in three-dimensional (3D) type I collagen matrices in vitro and in vivo. We hypothesized that human platelet lysate (HPL) would promote cell survival and enhance vasculogenesis in the 3D collagen matrices. We report that the percentage of ECFC co-cultured with HPL that were alive was significantly enhanced on days 1 and 3 post-matrix formation, compared to ECFC alone containing matrices. Also, co-culture of ECFC with HPL displayed significantly more vasculogenic activity compared to ECFC alone and expressed significantly more pro-survival molecules (pAkt, p-Bad and Bcl-xL) in the 3D collagen matrices in vitro. Treatment with Akt1 inhibitor (A-674563), Akt2 inhibitor (CCT128930) and Bcl-xL inhibitor (ABT-263/Navitoclax) significantly decreased the cell survival and vasculogenesis of ECFC co-cultured with or without HPL and implicated activation of the Akt1 pathway as the critical mediator of the HPL effect on ECFC in vitro. A significantly greater average vessel number and total vascular area of human CD31(+) vessels were present in implants containing ECFC and HPL, compared to the ECFC alone implants in vivo. We conclude that implantation of ECFC with HPL in vivo promotes vasculogenesis and augments blood vessel formation via diminishing apoptosis of the implanted ECFC

Angiopoietin-like protein 2 regulates endothelial colony forming cell vasculogenesis

Angiopoietin-like 2 (ANGPTL2) has been reported to induce sprouting angiogenesis; however, its role in vasculogenesis, the de novo lumenization of endothelial cells (EC), remains unexplored. We sought to investigate the potential role of ANGPTL2 in regulating human cord blood derived endothelial colony forming cell (ECFC) vasculogenesis through siRNA mediated inhibition of ANGPTL2 gene expression. We found that ECFCs in which ANGPTL2 was diminished displayed a threefold decrease in in vitro lumenal area whereas addition of exogenous ANGPTL2 protein domains to ECFCs lead to increased lumen formation within a 3 dimensional (3D) collagen assay of vasculogenesis. ECFC migration was attenuated by 36 % via ANGPTL2 knockdown (KD) although proliferation and apoptosis were not affected. We subsequently found that c-Jun NH2-terminal kinase (JNK), but not ERK1/2, phosphorylation was decreased upon ANGPTL2 KD, and expression of membrane type 1 matrix metalloproteinase (MT1-MMP), known to be regulated by JNK and a critical regulator of EC migration and 3D lumen formation, was decreased in lumenized structures in vitro derived from ANGPTL2 silenced ECFCs. Treatment of ECFCs in 3D collagen matrices with either a JNK inhibitor or exogenous rhTIMP-3 (an inhibitor of MT1-MMP activity) resulted in a similar phenotype of decreased vascular lumen formation as observed with ANGPTL2 KD, whereas stimulation of JNK activity increased vasculogenesis. Based on gene silencing, pharmacologic, cellular, and biochemical approaches, we conclude that ANGPTL2 positively regulates ECFC vascular lumen formation likely through its effects on migration and in part by activating JNK and increasing MT1-MMP expression

Notch ligand Delta-like 1 promotes in vivo vasculogenesis in human cord blood-derived endothelial colony forming cells

BACKGROUND AIMS: Human cord blood (CB) is enriched in circulating endothelial colony forming cells (ECFCs) that display high proliferative potential and in vivo vessel forming ability. Because Notch signaling is critical for embryonic blood vessel formation in utero, we hypothesized that Notch pathway activation may enhance cultured ECFC vasculogenic properties in vivo. METHODS: In vitro ECFC stimulation with an immobilized chimeric Notch ligand (Delta-like1(ext-IgG)) led to significant increases in the mRNA and protein levels of Notch regulated Hey2 and EphrinB2 that were blocked by treatment with γ-secretase inhibitor addition. However, Notch stimulated preconditioning in vitro failed to enhance ECFC vasculogenesis in vivo. In contrast, in vivo co-implantation of ECFCs with OP9-Delta-like 1 stromal cells that constitutively expressed the Notch ligand delta-like 1 resulted in enhanced Notch activated ECFC-derived increased vessel density and enlarged vessel area in vivo, an effect not induced by OP9 control stromal implantation. RESULTS: This Notch activation was associated with diminished apoptosis in the exposed ECFC. CONCLUSIONS: We conclude that Notch pathway activation in ECFC in vivo via co-implanted stromal cells expressing delta-like 1 promotes vasculogenesis and augments blood vessel formation via diminishing apoptosis of the implanted ECFC