A Newly Defined and Xeno-Free Culture Medium Supports Every-Other-Day Medium Replacement in the Generation and Long-Term Cultivation of Human Pluripotent Stem Cells.

PubMed

Ahmadian Baghbaderani, Behnam; Tian, Xinghui; Scotty Cadet, Jean; Shah, Kevan; Walde, Amy; Tran, Huan; Kovarcik, Don Paul; Clarke, Diana; Fellner, Thomas

2016-01-01

Human pluripotent stem cells (hPSCs) present an unprecedented opportunity to advance human health by offering an alternative and renewable cell resource for cellular therapeutics and regenerative medicine. The present demand for high quality hPSCs for use in both research and clinical studies underscores the need to develop technologies that will simplify the cultivation process and control variability. Here we describe the development of a robust, defined and xeno-free hPSC medium that supports reliable propagation of hPSCs and generation of human induced pluripotent stem cells (hiPSCs) from multiple somatic cell types; long-term serial subculturing of hPSCs with every-other-day (EOD) medium replacement; and banking fully characterized hPSCs. The hPSCs cultured in this medium for over 40 passages are genetically stable, retain high expression levels of the pluripotency markers TRA-1-60, TRA-1-81, Oct-3/4 and SSEA-4, and readily differentiate into ectoderm, mesoderm and endoderm. Importantly, the medium plays an integral role in establishing a cGMP-compliant process for the manufacturing of hiPSCs that can be used for generation of clinically relevant cell types for cell replacement therapy applications.

A Facile Method to Establish Human Induced Pluripotent Stem Cells From Adult Blood Cells Under Feeder-Free and Xeno-Free Culture Conditions: A Clinically Compliant Approach

PubMed Central

Chou, Bin-Kuan; Gu, Haihui; Gao, Yongxing; Dowey, Sarah N.; Wang, Ying; Shi, Jun; Li, Yanxin; Ye, Zhaohui; Cheng, Tao

2015-01-01

Reprogramming human adult blood mononuclear cells (MNCs) cells by transient plasmid expression is becoming increasingly popular as an attractive method for generating induced pluripotent stem (iPS) cells without the genomic alteration caused by genome-inserting vectors. However, its efficiency is relatively low with adult MNCs compared with cord blood MNCs and other fetal cells and is highly variable among different adult individuals. We report highly efficient iPS cell derivation under clinically compliant conditions via three major improvements. First, we revised a combination of three EBNA1/OriP episomal vectors expressing five transgenes, which increased reprogramming efficiency by ≥10–50-fold from our previous vectors. Second, human recombinant vitronectin proteins were used as cell culture substrates, alleviating the need for feeder cells or animal-sourced proteins. Finally, we eliminated the previously critical step of manually picking individual iPS cell clones by pooling newly emerged iPS cell colonies. Pooled cultures were then purified based on the presence of the TRA-1-60 pluripotency surface antigen, resulting in the ability to rapidly expand iPS cells for subsequent applications. These new improvements permit a consistent and reliable method to generate human iPS cells with minimal clonal variations from blood MNCs, including previously difficult samples such as those from patients with paroxysmal nocturnal hemoglobinuria. In addition, this method of efficiently generating iPS cells under feeder-free and xeno-free conditions allows for the establishment of clinically compliant iPS cell lines for future therapeutic applications. PMID:25742692

Long-term xeno-free culture of human pluripotent stem cells on hydrogels with optimal elasticity.

PubMed

Higuchi, Akon; Kao, Shih-Hsuan; Ling, Qing-Dong; Chen, Yen-Ming; Li, Hsing-Fen; Alarfaj, Abdullah A; Munusamy, Murugan A; Murugan, Kadarkarai; Chang, Shih-Chang; Lee, Hsin-Chung; Hsu, Shih-Tien; Kumar, S Suresh; Umezawa, Akihiro

2015-12-14

The tentative clinical application of human pluripotent stem cells (hPSCs), such as human embryonic stem cells and human induced pluripotent stem cells, is restricted by the possibility of xenogenic contamination resulting from the use of mouse embryonic fibroblasts (MEFs) as a feeder layer. Therefore, we investigated hPSC cultures on biomaterials with different elasticities that were grafted with different nanosegments. We prepared dishes coated with polyvinylalcohol-co-itaconic acid hydrogels grafted with an oligopeptide derived from vitronectin (KGGPQVTRGDVFTMP) with elasticities ranging from 10.3 to 30.4 kPa storage moduli by controlling the crosslinking time. The hPSCs cultured on the stiffest substrates (30.4 kPa) tended to differentiate after five days of culture, whereas the hPSCs cultured on the optimal elastic substrates (25 kPa) maintained their pluripotency for over 20 passages under xeno-free conditions. These results indicate that cell culture matrices with optimal elasticity can maintain the pluripotency of hPSCs in culture.

Culture in embryonic kidney serum and xeno-free media as renal cell carcinoma and renal cell carcinoma cancer stem cells research model.

PubMed

Krawczyk, Krzysztof M; Matak, Damian; Szymanski, Lukasz; Szczylik, Cezary; Porta, Camillo; Czarnecka, Anna M

2018-04-01

The use of fetal bovine serum hinders obtaining reproducible experimental results and should also be removed in hormone and growth factor studies. In particular hormones found in FBS act globally on cancer cell physiology and influence transcriptome and metabolome. The aim of our study was to develop a renal carcinoma serum free culture model optimized for (embryonal) renal cells in order to select the best study model for downstream auto-, para- or endocrine research. Secondary aim was to verify renal carcinoma stem cell culture for this application. In the study, we have cultured renal cell carcinoma primary tumour cell line (786-0) as well as human kidney cancer stem cells in standard 2D monolayer cultures in Roswell Park Memorial Institute Medium or Dulbecco's Modified Eagle's Medium and Complete Human Kidney Cancer Stem Cell Medium, respectively. Serum-free, animal-component free Human Embryonic Kidney 293 media were tested. Our results revealed that xeno-free embryonal renal cells optimized culture media provide a useful tool in RCC cancer biology research and at the same time enable effective growth of RCC. We propose bio-mimic RCC cell culture model with specific serum-free and xeno-free medium that promote RCC cell viability.

Xeno-free culture of human pluripotent stem cells on oligopeptide-grafted hydrogels with various molecular designs

PubMed Central

Chen, Yen-Ming; Chen, Li-Hua; Li, Meng-Pei; Li, Hsing-Fen; Higuchi, Akon; Kumar, S. Suresh; Ling, Qing-Dong; Alarfaj, Abdullah A.; Munusamy, Murugan A.; Chang, Yung; Benelli, Giovanni; Murugan, Kadarkarai; Umezawa, Akihiro

2017-01-01

Establishing cultures of human embryonic (ES) and induced pluripotent (iPS) stem cells in xeno-free conditions is essential for producing clinical-grade cells. Development of cell culture biomaterials for human ES and iPS cells is critical for this purpose. We designed several structures of oligopeptide-grafted poly (vinyl alcohol-co-itaconic acid) hydrogels with optimal elasticity, and prepared them in formations of single chain, single chain with joint segment, dual chain with joint segment, and branched-type chain. Oligopeptide sequences were selected from integrin- and glycosaminoglycan-binding domains of the extracellular matrix. The hydrogels grafted with vitronectin-derived oligopeptides having a joint segment or a dual chain, which has a storage modulus of 25 kPa, supported the long-term culture of human ES and iPS cells for over 10 passages. The dual chain and/or joint segment with cell adhesion molecules on the hydrogels facilitated the proliferation and pluripotency of human ES and iPS cells. PMID:28332572

Xeno-free culture of human pluripotent stem cells on oligopeptide-grafted hydrogels with various molecular designs.

PubMed

Chen, Yen-Ming; Chen, Li-Hua; Li, Meng-Pei; Li, Hsing-Fen; Higuchi, Akon; Kumar, S Suresh; Ling, Qing-Dong; Alarfaj, Abdullah A; Munusamy, Murugan A; Chang, Yung; Benelli, Giovanni; Murugan, Kadarkarai; Umezawa, Akihiro

2017-03-23

Establishing cultures of human embryonic (ES) and induced pluripotent (iPS) stem cells in xeno-free conditions is essential for producing clinical-grade cells. Development of cell culture biomaterials for human ES and iPS cells is critical for this purpose. We designed several structures of oligopeptide-grafted poly (vinyl alcohol-co-itaconic acid) hydrogels with optimal elasticity, and prepared them in formations of single chain, single chain with joint segment, dual chain with joint segment, and branched-type chain. Oligopeptide sequences were selected from integrin- and glycosaminoglycan-binding domains of the extracellular matrix. The hydrogels grafted with vitronectin-derived oligopeptides having a joint segment or a dual chain, which has a storage modulus of 25 kPa, supported the long-term culture of human ES and iPS cells for over 10 passages. The dual chain and/or joint segment with cell adhesion molecules on the hydrogels facilitated the proliferation and pluripotency of human ES and iPS cells.

Surface-engineered substrates for improved human pluripotent stem cell culture under fully defined conditions.

PubMed

Saha, Krishanu; Mei, Ying; Reisterer, Colin M; Pyzocha, Neena Kenton; Yang, Jing; Muffat, Julien; Davies, Martyn C; Alexander, Morgan R; Langer, Robert; Anderson, Daniel G; Jaenisch, Rudolf

2011-11-15

The current gold standard for the culture of human pluripotent stem cells requires the use of a feeder layer of cells. Here, we develop a spatially defined culture system based on UV/ozone radiation modification of typical cell culture plastics to define a favorable surface environment for human pluripotent stem cell culture. Chemical and geometrical optimization of the surfaces enables control of early cell aggregation from fully dissociated cells, as predicted from a numerical model of cell migration, and results in significant increases in cell growth of undifferentiated cells. These chemically defined xeno-free substrates generate more than three times the number of cells than feeder-containing substrates per surface area. Further, reprogramming and typical gene-targeting protocols can be readily performed on these engineered surfaces. These substrates provide an attractive cell culture platform for the production of clinically relevant factor-free reprogrammed cells from patient tissue samples and facilitate the definition of standardized scale-up friendly methods for disease modeling and cell therapeutic applications.

A simple, xeno-free method for oligodendrocyte generation from human neural stem cells derived from umbilical cord: engagement of gelatinases in cell commitment and differentiation.

PubMed

Sypecka, Joanna; Ziemka-Nalecz, Małgorzata; Dragun-Szymczak, Patrycja; Zalewska, Teresa

2017-05-01

Oligodendrocyte progenitors (OPCs) are ranked among the most likely candidates for cell-based strategies aimed at treating neurodegenerative diseases accompanied by dys/demyelination of the central nervous system (CNS). In this regard, different sources of stem cells are being tested to elaborate xeno-free protocols for efficient generation of OPCs for clinical applications. In the present study, neural stem cells of human umbilical cord blood (HUCB-NSCs) have been used to derive OPCs and subsequently to differentiate them into mature, GalC-expressing oligodendrocytes. Applied components of the extracellular matrix (ECM) and the analogues of physiological substances known to increase glial commitment of neural stem cells have been shown to significantly increase the yield of the resulting OPC fraction. The efficiency of ECM components in promoting oligodendrocyte commitment and differentiation prompted us to investigate the potential role of gelatinases in those processes. Subsequently, endogenous and ECM metalloproteinases (MMPs) activity has been compared with that detected in primary cultures of rat oligodendrocytes in vitro, as well as in rat brains in vivo. The data indicate that gelatinases are engaged in gliogenesis both in vitro and in vivo, although differently, which presumably results from distinct extracellular conditions. In conclusion, the study presents an efficient xeno-free method of deriving oligodendrocyte from HUCB-NSCs and analyses the engagement of MMP-2/MMP-9 in the processes of cell commitment and maturation. Copyright © 2015 John Wiley & Sons, Ltd. Copyright © 2015 John Wiley & Sons, Ltd.

Recombinant human type II collagen hydrogel provides a xeno-free 3D micro-environment for chondrogenesis of human bone marrow-derived mesenchymal stromal cells.

PubMed

Muhonen, Virpi; Narcisi, Roberto; Nystedt, Johanna; Korhonen, Matti; van Osch, Gerjo J V M; Kiviranta, Ilkka

2017-03-01

Recombinant human type II collagen (rhCII) hydrogel was tested as a xeno-free micro-environment for the chondrogenesis of human bone marrow-derived mesenchymal stromal cells (BM-MSCs). The rhCII hydrogels were seeded with BM-MSCs and cultured in a xeno-free chondro-inductive medium for 14, 28 and 84 days. High-density pellet cultures served as controls. The samples were subjected to biochemical, histological and gene expression analyses. Although the cells deposited glycosaminoglycans into the extracellular space significantly more slowly in the rhCII hydrogels compared to the high-density pellets, a similar potential of matrix deposition was reached by the end of the 84-day culture. At day 28 of culture, the gene expression level for cartilage marker genes (i.e. genes encoding for Sox9 transcription factor, Collagen type II and Aggrecan) were considerably lower in the rhCII hydrogels than in the high-density pellets, but at the end of the 84-day culture period, all the cartilage marker genes analysed were expressed at a similar level. Interestingly, the expression of the matrix metallopeptidases (MMP)-13, MMP-14 and MMP-8, i.e. extracellular collagen network-degrading enzymes, were transiently upregulated in the rhCII hydrogel, indicating active matrix reorganization. This study demonstrated that the rhCII hydrogel functions as a xeno-free platform for BM-MSC chondrogenesis, although the process is delayed. The reversible catabolic reaction evoked by the rhCII hydrogel might be beneficial in graft integration in vivo and pinpoints the need to further explore the use of hydrogels containing recombinant extracellular matrix (ECM) proteins to induce the chondrogenesis of MSCs. Copyright © 2015 John Wiley & Sons, Ltd. Copyright © 2015 John Wiley & Sons, Ltd.

Origin and significance of the 2011 El Hierro xeno-pumice

NASA Astrophysics Data System (ADS)

Zaczek-Pedroza, Kirsten; Troll, Valentin R.; Deegan, Frances M.; Meade, Fiona C.; Burchardt, Steffi; Carracedo, Juan C.; Klügel, Andreas; Harris, Chris; Wiesmaier, Sebastian; Berg, Sylvia E.; Barker, Abigail K.

2014-05-01

During the first week of the 2011/2012 submarine eruption off El Hierro (Canary Islands), peculiar light-coloured pumiceous rocks (xeno-pumice) were found floating on the sea. The appearance of these rocks led to a potentially inappropriate response from the authorities, because the rocks were viewed as likely indicators of high-silica magma and possible explosive eruptive behaviour. However, the eruption remained a relatively minor and dominantly effusive event and the origin and significance of these peculiar xeno-pumice rocks for volcanic monitoring remains unresolved. Three contrasting models have been put forward, describing them as: a) recycled hydrothermally altered felsic magmatic rocks (Meletlidis et al., 2012, Geophys. Res. Lett., 39:L17302), b) sediment-contaminated high-silica magma (Sigmarsson et al., 2013, Contrib. Mineral. Petrol., 165:601-622) or c) frothy xenoliths from pre-island sedimentary strata that were melted while immersed in magma (Troll et al., 2012, Solid Earth, 3:97-110). Here, we combine the three available datasets to assess the origin of El Hierro xeno-pumice in the light of texture, mineralogy, major and trace element composition, and oxygen isotope characteristics in order to discuss their significance. We note that: 1) Sedimentary relicts occur frequently in xeno-pumice samples with occasionally observed relict bedding. 2) Vesicle sizes are extremely variable, which documents multiple degassing events. The vesicles are biggest especially close to sedimentary relicts, likely the result of a complex mix of minerals and porewaters originally present. 3) The mineral assemblage of xeno-pumice includes quartz, smectite, illite, wollastonite, jasper and mica (XRD) and is akin to marine sedimentary rocks in the region (Hoernle, 1998, J. Petrol.,39:859-880; Robertson & Stillman, 1979, J. Geol. Soc., 136:47 -60; Aparicio et al., 2006, Geol. Mag. 143:181 -193). 4) CIPW norms calculated from xeno-pumice major element compositions show the

Human serum-derived protein removes the need for coating in defined human pluripotent stem cell culture

PubMed Central

Pijuan-Galitó, Sara; Tamm, Christoffer; Schuster, Jens; Sobol, Maria; Forsberg, Lars; Merry, Catherine L. R.; Annerén, Cecilia

2016-01-01

Reliable, scalable and time-efficient culture methods are required to fully realize the clinical and industrial applications of human pluripotent stem (hPS) cells. Here we present a completely defined, xeno-free medium that supports long-term propagation of hPS cells on uncoated tissue culture plastic. The medium consists of the Essential 8 (E8) formulation supplemented with inter-α-inhibitor (IαI), a human serum-derived protein, recently demonstrated to activate key pluripotency pathways in mouse PS cells. IαI efficiently induces attachment and long-term growth of both embryonic and induced hPS cell lines when added as a soluble protein to the medium at seeding. IαI supplementation efficiently supports adaptation of feeder-dependent hPS cells to xeno-free conditions, clonal growth as well as single-cell survival in the absence of Rho-associated kinase inhibitor (ROCKi). This time-efficient and simplified culture method paves the way for large-scale, high-throughput hPS cell culture, and will be valuable for both basic research and commercial applications. PMID:27405751

XenoSite server: a web-available site of metabolism prediction tool.

PubMed

Matlock, Matthew K; Hughes, Tyler B; Swamidass, S Joshua

2015-04-01

Cytochrome P450 enzymes (P450s) are metabolic enzymes that process the majority of FDA-approved, small-molecule drugs. Understanding how these enzymes modify molecule structure is key to the development of safe, effective drugs. XenoSite server is an online implementation of the XenoSite, a recently published computational model for P450 metabolism. XenoSite predicts which atomic sites of a molecule--sites of metabolism (SOMs)--are modified by P450s. XenoSite server accepts input in common chemical file formats including SDF and SMILES and provides tools for visualizing the likelihood that each atomic site is a site of metabolism for a variety of important P450s, as well as a flat file download of SOM predictions. XenoSite server is available at http://swami.wustl.edu/xenosite. © The Author 2014. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

An optimal serum-free defined condition for in vitro culture of kidney organoids.

PubMed

Nishikawa, Masaki; Kimura, Hiroshi; Yanagawa, Naomi; Hamon, Morgan; Hauser, Peter; Zhao, Lifu; Jo, Oak D; Yanagawa, Norimoto

2018-07-02

Kidney organoid is an emerging topic of importance for research in kidney development and regeneration. Conventional culture systems for kidney organoids reported thus far use culture media containing serum, which may compromise our understanding and the potential clinical applicability of the organoid system. In our present study, we tested two serum-free culture conditions and compared their suitability for the maintenance and growth of kidney organoids in culture. One of the serum-free culture conditions was the combination of keratinocytes serum free medium (KSFM) with knockout serum replacement (KSR) (KSFM + KSR), and the other was the combination of knockout DMEM/F12 (KD/F12) and KSR (KD/F12 + KSR). With cell aggregates derived from E12.5 mouse embryonic kidneys, we found that KD/F12 + KSR was superior to KSFM + KSR in promoting the growth of the aggregate with expansion of Six2 + nephron progenitor cells (NPC) and elaborated ureteric branching morphogenesis. With KD/F12 + KSR, we found that lower concentrations of KSR at 5-10% were superior to a higher concentration (20%) in promoting the growth of aggregates without affecting the expression levels of NPC marker genes. We also found that NPC in aggregates retained their differentiation potential to develop nephron tubules through mesenchyme-to-epithelial transition (MET), after being maintained in culture under these conditions for up to 7 days. In conclusion, we have identified a defined serum-free culture condition suitable for the maintenance and growth of kidney organoids that retain the differentiation potential to develop nephron structures. This defined serum-free culture condition may serve as a useful platform for further investigation of kidney organoids in vitro. Copyright © 2018 Elsevier Inc. All rights reserved.

Optimization of Pre-transplantation Conditions to Enhance the Efficacy of Mesenchymal Stem Cells

PubMed Central

Haque, Nazmul; Kasim, Noor Hayaty Abu; Rahman, Mohammad Tariqur

2015-01-01

Mesenchymal stem cells (MSCs) are considered a potential tool for cell based regenerative therapy due to their immunomodulatory property, differentiation potentials, trophic activity as well as large donor pool. Poor engraftment and short term survival of transplanted MSCs are recognized as major limitations which were linked to early cellular ageing, loss of chemokine markers during ex vivo expansion, and hyper-immunogenicity to xeno-contaminated MSCs. These problems can be minimized by ex vivo expansion of MSCs in hypoxic culture condition using well defined or xeno-free media i.e., media supplemented with growth factors, human serum or platelet lysate. In addition to ex vivo expansion in hypoxic culture condition using well defined media, this review article describes the potentials of transient adaptation of expanded MSCs in autologous serum supplemented medium prior to transplantation for long term regenerative benefits. Such transient adaptation in autologous serum supplemented medium may help to increase chemokine receptor expression and tissue specific differentiation of ex vivo expanded MSCs, thus would provide long term regenerative benefits. PMID:25678851

Optimized human platelet lysate as novel basis for a serum-, xeno-, and additive-free corneal endothelial cell and tissue culture.

PubMed

Thieme, Daniel; Reuland, Lynn; Lindl, Toni; Kruse, Friedrich; Fuchsluger, Thomas

2018-02-01

The expansion of donor-derived corneal endothelial cells (ECs) is a promising approach for regenerative therapies in corneal diseases. To achieve the best Good Manufacturing Practice standard the entire cultivation process should be devoid of nonhuman components. However, so far, there is no suitable xeno-free protocol for clinical applications. We therefore introduce a processed variant of a platelet lysate for the use in corneal cell and tissue culture based on a Good Manufacturing Practice-grade thrombocyte concentrate. This processed human platelet lysate (phPL), free of any animal components and of anticoagulants such as heparin with a physiological ionic composition, was used to cultivate corneal ECs in vitro and ex vivo in comparison to standard cultivation with fetal calf serum (FCS). Human donor corneas were cut in quarters while 2 quarters of each cornea were incubated with the respective medium supplement. Three fields of view per quarter were taken into account for the analysis. Evaluation of phPL as a medium supplement in cell culture of immortalized EC showed a superior viability compared with FCS control with reduced cell proliferation. Furthermore, the viability during the expansion of primary cells is significantly (3-fold ±0.5) increased with phPL compared with FCS standard medium. Quartering donor corneas was traumatic for the endothelium and therefore resulted in increased EC loss. Interestingly, however, cultivation of the quartered pieces for 2 weeks in 0.1-mg/ml pHPL in Biochrome I showed a 21 (±10) % EC loss compared with 67 (±12) % EC loss when cultivated in 2% FCS in Biochrome I. The cell culture protocol with pHPL as FCS replacement seems to be superior to the standard FCS protocols with respect to EC survival. It offers a xeno-free and physiological environment for corneal endothelial cells. This alternative cultivation protocol could facilitate the use of EC for human corneal cell therapy. Copyright © 2017 John Wiley & Sons, Ltd.

Establishment of feeder-free culture system for human induced pluripotent stem cell on DAS nanocrystalline graphene

NASA Astrophysics Data System (ADS)

Lee, Hyunah; Nam, Donggyu; Choi, Jae-Kyung; Araúzo-Bravo, Marcos J.; Kwon, Soon-Yong; Zaehres, Holm; Lee, Taehee; Park, Chan Young; Kang, Hyun-Wook; Schöler, Hans R.; Kim, Jeong Beom

2016-02-01

The maintenance of undifferentiated human pluripotent stem cells (hPSC) under xeno-free condition requires the use of human feeder cells or extracellular matrix (ECM) coating. However, human-derived sources may cause human pathogen contamination by viral or non-viral agents to the patients. Here we demonstrate feeder-free and xeno-free culture system for hPSC expansion using diffusion assisted synthesis-grown nanocrystalline graphene (DAS-NG), a synthetic non-biological nanomaterial which completely rule out the concern of human pathogen contamination. DAS-NG exhibited advanced biocompatibilities including surface nanoroughness, oxygen containing functional groups and hydrophilicity. hPSC cultured on DAS-NG could maintain pluripotency in vitro and in vivo, and especially cell adhesion-related gene expression profile was comparable to those of cultured on feeders, while hPSC cultured without DAS-NG differentiated spontaneously with high expression of somatic cell-enriched adhesion genes. This feeder-free and xeno-free culture method using DAS-NG will facilitate the generation of clinical-grade hPSC.

Cryopreserved, Xeno-Free Human Umbilical Cord Mesenchymal Stromal Cells Reduce Lung Injury Severity and Bacterial Burden in Rodent Escherichia coli-Induced Acute Respiratory Distress Syndrome.

PubMed

Curley, Gerard F; Jerkic, Mirjana; Dixon, Steve; Hogan, Grace; Masterson, Claire; O'Toole, Daniel; Devaney, James; Laffey, John G

2017-02-01

Although mesenchymal stem/stromal cells represent a promising therapeutic strategy for acute respiratory distress syndrome, clinical translation faces challenges, including scarcity of bone marrow donors, and reliance on bovine serum during mesenchymal stem/stromal cell proliferation. We wished to compare mesenchymal stem/stromal cells from human umbilical cord, grown in xeno-free conditions, with mesenchymal stem/stromal cells from human bone marrow, in a rat model of Escherichia coli pneumonia. In addition, we wished to determine the potential for umbilical cord-mesenchymal stem/stromal cells to reduce E. coli-induced oxidant injury. Randomized animal study. University research laboratory. Male Sprague-Dawley rats. Acute respiratory distress syndrome was induced in rats by intratracheal instillation of E. coli (1.5-2 × 10 CFU/kg). "Series 1" compared the effects of freshly thawed cryopreserved umbilical cord-mesenchymal stem/stromal cells with bone marrow-mesenchymal stem/stromal cells on physiologic indices of lung injury, cellular infiltration, and E. coli colony counts in bronchoalveolar lavage. "Series 2" examined the effects of cryopreserved umbilical cord-mesenchymal stem/stromal cells on survival, as well as measures of injury, inflammation and oxidant stress, including production of reactive oxidative species, reactive oxidative species scavenging by superoxide dismutase-1 and superoxide dismutase-2. In "Series 1," animals subjected to E. coli pneumonia who received umbilical cord-mesenchymal stem/stromal cells had improvements in oxygenation, respiratory static compliance, and wet-to-dry ratios comparable to bone marrow-mesenchymal stem/stromal cell treatment. E. coli colony-forming units in bronchoalveolar lavage were reduced in both cell therapy groups, despite a reduction in bronchoalveolar lavage neutrophils. In series 2, umbilical cord-mesenchymal stem/stromal cells enhanced animal survival and decreased alveolar protein and proinflammatory

Non-integrating episomal plasmid-based reprogramming of human amniotic fluid stem cells into induced pluripotent stem cells in chemically defined conditions.

PubMed

Slamecka, Jaroslav; Salimova, Lilia; McClellan, Steven; van Kelle, Mathieu; Kehl, Debora; Laurini, Javier; Cinelli, Paolo; Owen, Laurie; Hoerstrup, Simon P; Weber, Benedikt

2016-01-01

Amniotic fluid stem cells (AFSC) represent an attractive potential cell source for fetal and pediatric cell-based therapies. However, upgrading them to pluripotency confers refractoriness toward senescence, higher proliferation rate and unlimited differentiation potential. AFSC were observed to rapidly and efficiently reacquire pluripotency which together with their easy recovery makes them an attractive cell source for reprogramming. The reprogramming process as well as the resulting iPSC epigenome could potentially benefit from the unspecialized nature of AFSC. iPSC derived from AFSC also have potential in disease modeling, such as Down syndrome or β-thalassemia. Previous experiments involving AFSC reprogramming have largely relied on integrative vector transgene delivery and undefined serum-containing, feeder-dependent culture. Here, we describe non-integrative oriP/EBNA-1 episomal plasmid-based reprogramming of AFSC into iPSC and culture in fully chemically defined xeno-free conditions represented by vitronectin coating and E8 medium, a system that we found uniquely suited for this purpose. The derived AF-iPSC lines uniformly expressed a set of pluripotency markers Oct3/4, Nanog, Sox2, SSEA-1, SSEA-4, TRA-1-60, TRA-1-81 in a pattern typical for human primed PSC. Additionally, the cells formed teratomas, and were deemed pluripotent by PluriTest, a global expression microarray-based in-silico pluripotency assay. However, we found that the PluriTest scores were borderline, indicating a unique pluripotent signature in the defined condition. In the light of potential future clinical translation of iPSC technology, non-integrating reprogramming and chemically defined culture are more acceptable.

XenoSite: accurately predicting CYP-mediated sites of metabolism with neural networks.

PubMed

Zaretzki, Jed; Matlock, Matthew; Swamidass, S Joshua

2013-12-23

Understanding how xenobiotic molecules are metabolized is important because it influences the safety, efficacy, and dose of medicines and how they can be modified to improve these properties. The cytochrome P450s (CYPs) are proteins responsible for metabolizing 90% of drugs on the market, and many computational methods can predict which atomic sites of a molecule--sites of metabolism (SOMs)--are modified during CYP-mediated metabolism. This study improves on prior methods of predicting CYP-mediated SOMs by using new descriptors and machine learning based on neural networks. The new method, XenoSite, is faster to train and more accurate by as much as 4% or 5% for some isozymes. Furthermore, some "incorrect" predictions made by XenoSite were subsequently validated as correct predictions by revaluation of the source literature. Moreover, XenoSite output is interpretable as a probability, which reflects both the confidence of the model that a particular atom is metabolized and the statistical likelihood that its prediction for that atom is correct.

Heterogeneous vesiculation of 2011 El Hierro xeno-pumice revealed by X-ray computed microtomography

NASA Astrophysics Data System (ADS)

Berg, S. E.; Troll, V. R.; Deegan, F. M.; Burchardt, S.; Krumbholz, M.; Mancini, L.; Polacci, M.; Carracedo, J. C.; Soler, V.; Arzilli, F.; Brun, F.

2016-12-01

During the first week of the 2011 El Hierro submarine eruption, abundant light-coloured pumiceous, high-silica volcanic bombs coated in dark basanite were found floating on the sea. The composition of the light-coloured frothy material (`xeno-pumice') is akin to that of sedimentary rocks from the region, but the textures resemble felsic magmatic pumice, leaving their exact mode of formation unclear. To help decipher their origin, we investigated representative El Hierro xeno-pumice samples using X-ray computed microtomography for their internal vesicle shapes, volumes, and bulk porosity, as well as for the spatial arrangement and size distributions of vesicles in three dimensions (3D). We find a wide range of vesicle morphologies, which are especially variable around small fragments of rock contained in the xeno-pumice samples. Notably, these rock fragments are almost exclusively of sedimentary origin, and we therefore interpret them as relicts an the original sedimentary ocean crust protolith(s). The irregular vesiculation textures observed probably resulted from pulsatory release of volatiles from multiple sources during xeno-pumice formation, most likely by successive release of pore water and mineral water during incremental heating and decompression of the sedimentary protoliths.

Top-Down Inhibition of BMP Signaling Enables Robust Induction of hPSCs Into Neural Crest in Fully Defined, Xeno-free Conditions.

PubMed

Hackland, James O S; Frith, Tom J R; Thompson, Oliver; Marin Navarro, Ana; Garcia-Castro, Martin I; Unger, Christian; Andrews, Peter W

2017-10-10

Defects in neural crest development have been implicated in many human disorders, but information about human neural crest formation mostly depends on extrapolation from model organisms. Human pluripotent stem cells (hPSCs) can be differentiated into in vitro counterparts of the neural crest, and some of the signals known to induce neural crest formation in vivo are required during this process. However, the protocols in current use tend to produce variable results, and there is no consensus as to the precise signals required for optimal neural crest differentiation. Using a fully defined culture system, we have now found that the efficient differentiation of hPSCs to neural crest depends on precise levels of BMP signaling, which are vulnerable to fluctuations in endogenous BMP production. We present a method that controls for this phenomenon and could be applied to other systems where endogenous signaling can also affect the outcome of differentiation protocols. Copyright © 2017 The Authors. Published by Elsevier Inc. All rights reserved.

Defined, serum/feeder-free conditions for expansion and drug screening of primary B-acute lymphoblastic leukemia.

PubMed

Jiang, Zhiwu; Wu, Di; Ye, Wei; Weng, Jianyu; Lai, Peilong; Shi, Pengcheng; Guo, Xutao; Huang, Guohua; Deng, Qiuhua; Tang, Yanlai; Zhao, Hongyu; Cui, Shuzhong; Lin, Simiao; Wang, Suna; Li, Baiheng; Wu, Qiting; Li, Yangqiu; Liu, Pentao; Pei, Duanqing; Du, Xin; Yao, Yao; Li, Peng

2017-12-05

Functional screening for compounds represents a major hurdle in the development of rational therapeutics for B-acute lymphoblastic leukemia (B-ALL). In addition, using cell lines as valid models for evaluating responses to novel drug therapies raises serious concerns, as cell lines are prone to genotypic/phenotypic drift and loss of heterogeneity in vitro . Here, we reported that OP9 cells, not OP9-derived adipocytes (OP9TA), support the growth of primary B-ALL cells in vitro . To identify the factors from OP9 cells that support the growth of primary B-ALL cells, we performed RNA-Seq to analyze the gene expression profiles of OP9 and OP9TA cells. We thus developed a defined, serum/feeder-free condition (FI76V) that can support the expansion of a range of clinically distinct primary B-ALL cells that still maintain their leukemia-initiating ability. We demonstrated the suitability of high-throughput drug screening based on our B-ALL cultured conditions. Upon screening 378 kinase inhibitors, we identified a cluster of 17 kinase inhibitors that can efficiently kill B-ALL cells in vitro . Importantly, we demonstrated the synergistic cytotoxicity of dinaciclib/BTG226 to B-ALL cells. Taken together, we developed a defined condition for the ex vivo expansion of primary B-ALL cells that is suitable for high-throughput screening of novel compounds.

Human serum and platelet lysate are appropriate xeno-free alternatives for clinical-grade production of human MuStem cell batches.

PubMed

Saury, Charlotte; Lardenois, Aurélie; Schleder, Cindy; Leroux, Isabelle; Lieubeau, Blandine; David, Laurent; Charrier, Marine; Guével, Laëtitia; Viau, Sabrina; Delorme, Bruno; Rouger, Karl

2018-05-02

Canine MuStem cells have demonstrated regenerative efficacy in a dog model of muscular dystrophy, and the recent characterization of human counterparts (hMuStem) has highlighted the therapeutic potential of this muscle-derived stem cell population. To date, these cells have only been generated in research-grade conditions. However, evaluation of the clinical efficacy of any such therapy will require the production of hMuStem cells in compliance with good manufacturing practices (GMPs). Because the current use of fetal bovine serum (FBS) to isolate and expand hMuStem cells raises several ethical, safety, and supply concerns, we assessed the use of two alternative xeno-free blood derivatives: human serum (HS) and a human platelet lysate (hPL). hMuStem cells were isolated and expanded in vitro in either HS-supplemented or hPL-supplemented media and the proliferation rate, clonogenicity, myogenic commitment potential, and oligopotency compared with that observed in FBS-supplemented medium. Flow cytometry and high-throughput 3'-digital gene expression RNA sequencing were used to characterize the phenotype and global gene expression pattern of hMuStem cells cultured with HS or hPL. HS-supplemented and hPL-supplemented media both supported the isolation and long-term proliferation of hMuStem cells. Compared with FBS-based medium, both supplements enhanced clonogenicity and allowed for a reduction in growth factor supplementation. Neither supplement altered the cell lineage pattern of hMuStem cells. In vitro differentiation assays revealed a decrease in myogenic commitment and in the fusion ability of hMuStem cells when cultured with hPL. In return, this reduction of myogenic potential in hPL-supplemented cultures was rapidly reversed by substitution of hPL with HS or fibrinogen-depleted hPL. Moreover, culture of hMuStem cells in hPL hydrogel and fibrinogen-depleted hPL demonstrated that myogenic differentiation potential is maintained in heparin-free hPL derivatives. Our

Stirred tank bioreactor culture combined with serum-/xenogeneic-free culture medium enables an efficient expansion of umbilical cord-derived mesenchymal stem/stromal cells.

PubMed

Mizukami, Amanda; Fernandes-Platzgummer, Ana; Carmelo, Joana G; Swiech, Kamilla; Covas, Dimas T; Cabral, Joaquim M S; da Silva, Cláudia L

2016-08-01

Mesenchymal stem/stromal cells (MSC) are being widely explored as promising candidates for cell-based therapies. Among the different human MSC origins exploited, umbilical cord represents an attractive and readily available source of MSC that involves a non-invasive collection procedure. In order to achieve relevant cell numbers of human MSC for clinical applications, it is crucial to develop scalable culture systems that allow bioprocess control and monitoring, combined with the use of serum/xenogeneic (xeno)-free culture media. In the present study, we firstly established a spinner flask culture system combining gelatin-based Cultispher(®) S microcarriers and xeno-free culture medium for the expansion of umbilical cord matrix (UCM)-derived MSC. This system enabled the production of 2.4 (±1.1) x10(5) cells/mL (n = 4) after 5 days of culture, corresponding to a 5.3 (±1.6)-fold increase in cell number. The established protocol was then implemented in a stirred-tank bioreactor (800 mL working volume) (n = 3) yielding 115 million cells after 4 days. Upon expansion under stirred conditions, cells retained their differentiation ability and immunomodulatory potential. The development of a scalable microcarrier-based stirred culture system, using xeno-free culture medium that suits the intrinsic features of UCM-derived MSC represents an important step towards a GMP compliant large-scale production platform for these promising cell therapy candidates. Copyright © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Kidney regeneration by xeno-embryonic nephrogenesis.

PubMed

Yokoo, Takashi; Fukui, Akira; Matsumoto, Kei; Kawamura, Tetsuya

2008-03-01

Establishment of a functional whole kidney de novo has not received much attention because of the formidable challenges and the slow pace of advances in this field of research. This situation has changed recently with publication of data revealing the catastrophic nature of Medicaid costs for dialysis-related diseases. An innovative approach is needed in our search for therapies for kidney diseases and to provide a substitute for dialysis as soon as possible. Regenerative medicine offers great hope for realizing this goal. We established a system by which human mesenchymal stem cells can differentiate into a functional renal unit using a program of nephrogenesis in a developing xeno-embryo. In this article, recent research in the field of developing whole kidneys is reviewed, and possible therapeutic applications for kidney diseases are proposed in combination with our knowledge of the emerging field of kidney stem cell biology.

Isolation and expansion of human pluripotent stem cell-derived hepatic progenitor cells by growth factor defined serum-free culture conditions.

PubMed

Fukuda, Takayuki; Takayama, Kazuo; Hirata, Mitsuhi; Liu, Yu-Jung; Yanagihara, Kana; Suga, Mika; Mizuguchi, Hiroyuki; Furue, Miho K

2017-03-15

Limited growth potential, narrow ranges of sources, and difference in variability and functions from batch to batch of primary hepatocytes cause a problem for predicting drug-induced hepatotoxicity during drug development. Human pluripotent stem cell (hPSC)-derived hepatocyte-like cells in vitro are expected as a tool for predicting drug-induced hepatotoxicity. Several studies have already reported efficient methods for differentiating hPSCs into hepatocyte-like cells, however its differentiation process is time-consuming, labor-intensive, cost-intensive, and unstable. In order to solve this problem, expansion culture for hPSC-derived hepatic progenitor cells, including hepatic stem cells and hepatoblasts which can self-renewal and differentiate into hepatocytes should be valuable as a source of hepatocytes. However, the mechanisms of the expansion of hPSC-derived hepatic progenitor cells are not yet fully understood. In this study, to isolate hPSC-derived hepatic progenitor cells, we tried to develop serum-free growth factor defined culture conditions using defined components. Our culture conditions were able to isolate and grow hPSC-derived hepatic progenitor cells which could differentiate into hepatocyte-like cells through hepatoblast-like cells. We have confirmed that the hepatocyte-like cells prepared by our methods were able to increase gene expression of cytochrome P450 enzymes upon encountering rifampicin, phenobarbital, or omeprazole. The isolation and expansion of hPSC-derived hepatic progenitor cells in defined culture conditions should have advantages in terms of detecting accurate effects of exogenous factors on hepatic lineage differentiation, understanding mechanisms underlying self-renewal ability of hepatic progenitor cells, and stably supplying functional hepatic cells. Copyright © 2017 The Authors. Published by Elsevier Inc. All rights reserved.

Scalable cultivation of human pluripotent stem cells on chemically-defined surfaces

NASA Astrophysics Data System (ADS)

Hsiung, Michael Chi-Wei

Human stem cells (SCs) are classified as self-renewing cells possessing great ability in therapeutic applications due of their ability to differentiate along any major cell lineage in the human body. Despite their restorative potential, widespread use of SCs is hampered by strenuous control issues. Along with the need for strict xeno-free environments to sustain growth in culture, current methods for growing human pluripotent stem cells (hPSCs) rely on platforms which impede large-scale cultivation and therapeutic delivery. Hence, any progress towards development of large-scale culture systems is severely hindered. In a concentrated effort to develop a scheme that can serve as a model precursor for large scale SC propagation in clinical use, we have explored methods for cultivating hPSCs on completely defined surfaces. We discuss novel approaches with the potential to go beyond the limitations presented by current methods. In particular, we studied the cultivation of human embryonic stem cells (hESCs) and human induced pluripotent stem cells (hiPSCs) on surface which underwent synthetic or chemical modification. Current methods for hPSCs rely on animal-based extracellular matrices (ECMs) such as mouse embryonic fibroblasts (MEFs) or feeders and murine sacoma cell-derived substrates to facilitate their growth. While these layers or coatings can be used to maximize the output of hPSC production, they cannot be considered for clinical use because they risk introducing foreign pathogens into culture. We have identified and developed conditions for a completely defined xeno-free substrate used for culturing hPSCs. By utilizing coupling chemistry, we can functionalize ester groups on a given surface and conjugate synthetic peptides containing the arginine-glycine-aspartic acid (RGD) motif, known for their role in cell adhesion. This method offers advantages over traditional hPSC culture by keeping the modified substrata free of xenogenic response and can be scaled up in

Chemically defined serum-free conditions for cartilage regeneration from human embryonic stem cells.

PubMed

Yang, Dandan; Chen, Shubin; Gao, Changzhao; Liu, Xiaobo; Zhou, Yulai; Liu, Pengfei; Cai, Jinglei

2016-11-01

The aim of this study was to improve a method that induce cartilage differentiation of human embryoid stem cells (hESCs) in vitro, and test the effect of in vivo environments on the further maturation of hESCs derived cells. Embryoid bodies (EBs) formed from hESCs, with serum-free KSR-based medium and mesodermal specification related factors, CHIR, and Noggin for first 8days. Then cells were digested and cultured as micropellets in serum-free KSR-based chondrogenic medium that was supplemented with PDGF-BB, TGF β3, BMP4 in sequence for 24days. The morphology, FACS, histological staining as well as the expression of chondrogenic specific genes were detected in each stage, and further in vivo experiments, cell injections and tissue transplantations, further verified the formation of chondrocytes. We were able to obtain chondrocyte/cartilage from hESCs using serum-free KSR-based conditioned medium. qPCR analysis showed that expression of the chondroprogenitor genes and the chondrocyte/cartilage matrix genes. Morphology analysis demonstrated we got PG+COL2+COL1-particles. It indicated we obtained hyaline cartilage-like particles. 32-Day differential cells were injected subcutaneous. Staining results showed grafts developed further mature in vivo. But when transplanted in subrenal capsule, their effect was not good as in subcutaneous. Microenvironment might affect the cartilage formation. The results of this study provide an absolute serum-free and efficient approach for generation of hESC-derived chondrocytes, and cells will become further maturation in vivo. It provides evidence and technology for the hypothesis that hESCs may be a promising therapy for the treatment of cartilage disease. Copyright © 2016 Elsevier Inc. All rights reserved.

Norepinephrine stimulates progesterone production in highly estrogenic bovine granulosa cells cultured under serum-free, chemically defined conditions.

PubMed

Piccinato, Carla A; Montrezor, Luis H; Collares, Cristhianna A V; Vireque, Alessandra A; Rosa e Silva, Alzira A M

2012-11-22

Since noradrenergic innervation was described in the ovarian follicle, the actions of the intraovarian catecholaminergic system have been the focus of a variety of studies. We aimed to determine the gonadotropin-independent effects of the catecholamine norepinephrine (NE) in the steroid hormone profile of a serum-free granulosa cell (GC) culture system in the context of follicular development and dominance. Primary bovine GCs were cultivated in a serum-free, chemically defined culture system supplemented with 0.1% polyvinyl alcohol. The culture features were assessed by hormone measurements and ultrastructural characteristics of GCs. GCs produced increasing amounts of estradiol and pregnenolone for 144h and maintained ultrastructural features of healthy steroidogenic cells. Progesterone production was also detected, although it significantly increased only after 96h of culture. There was a highly significant positive correlation between estradiol and pregnenolone production in high E2-producing cultures. The effects of NE were further evaluated in a dose-response study. The highest tested concentration of NE (10 (-7) M) resulted in a significant increase in progesterone production, but not in estradiol or pregnenolone production. The specificity of NE effects on progesterone production was further investigated by incubating GCs with propranolol (10 (-8) M), a non-selective beta-adrenergic antagonist. The present culture system represents a robust model to study the impact of intrafollicular factors, such as catecholamines, in ovarian steroidogenesis and follicular development. The results of noradrenergic effects in the steroidogenesis of GC have implications on physiological follicular fate and on certain pathological ovarian conditions such as cyst formation and anovulation.

Discovery and bio-optimization of human antibody therapeutics using the XenoMouse® transgenic mouse platform.

PubMed

Foltz, Ian N; Gunasekaran, Kannan; King, Chadwick T

2016-03-01

Since the late 1990s, the use of transgenic animal platforms has transformed the discovery of fully human therapeutic monoclonal antibodies. The first approved therapy derived from a transgenic platform--the epidermal growth factor receptor antagonist panitumumab to treat advanced colorectal cancer--was developed using XenoMouse(®) technology. Since its approval in 2006, the science of discovering and developing therapeutic monoclonal antibodies derived from the XenoMouse(®) platform has advanced considerably. The emerging array of antibody therapeutics developed using transgenic technologies is expected to include antibodies and antibody fragments with novel mechanisms of action and extreme potencies. In addition to these impressive functional properties, these antibodies will be designed to have superior biophysical properties that enable highly efficient large-scale manufacturing methods. Achieving these new heights in antibody drug discovery will ultimately bring better medicines to patients. Here, we review best practices for the discovery and bio-optimization of monoclonal antibodies that fit functional design goals and meet high manufacturing standards. © 2016 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

Culture of human mesenchymal stem cells using a candidate pharmaceutical grade xeno-free cell culture supplement derived from industrial human plasma pools.

PubMed

Díez, José M; Bauman, Ewa; Gajardo, Rodrigo; Jorquera, Juan I

2015-03-13

Fetal bovine serum (FBS) is an animal product used as a medium supplement. The animal origin of FBS is a concern if cultured stem cells are to be utilized for human cell therapy. Therefore, a substitute for FBS is desirable. In this study, an industrial, xeno-free, pharmaceutical-grade supplement for cell culture (SCC) under development at Grifols was tested for growth of human mesenchymal stem cells (hMSCs), cell characterization, and differentiation capacity. SCC is a freeze-dried product obtained through cold-ethanol fractionation of industrial human plasma pools from healthy donors. Bone marrow-derived hMSC cell lines were obtained from two commercial suppliers. Cell growth was evaluated by culturing hMSCs with commercial media or media supplemented with SCC or FBS. Cell viability and cell yield were assessed with an automated cell counter. Cell surface markers were studied by indirect immunofluorescence assay. Cells were cultured then differentiated into adipocytes, chondrocytes, osteoblasts, and neurons, as assessed by specific staining and microscopy observation. SCC supported the growth of commercial hMSCs. Starting from the same number of seeded cells in two consecutive passages of culture with medium supplemented with SCC, hMSC yield and cell population doubling time were equivalent to the values obtained with the commercial medium and was consistent among lots. The viability of hMSCs was higher than 90%, while maintaining the characteristic phenotype of undifferentiated hMSCs (positive for CD29, CD44, CD90, CD105, CD146, CD166 and Stro-1; negative for CD14 and CD19). Cultured hMSCs maintained the potential for differentiation into adipocytes, chondrocytes, osteoblasts, and neurons. The tested human plasma-derived SCC sustains the adequate growth of hMSCs, while preserving their differentiation capacity. SCC can be a potential candidate for cell culture supplement in advanced cell therapies.

Generation of enhanced definitive endoderm from human embryonic stem cells under an albumin/insulin-free and chemically defined condition.

PubMed

Qu, Su; Yan, Liang; Fang, Bo; Ye, Shoudong; Li, Ping; Ge, Shengyang; Wu, Jian; Qu, Di; Song, Houyan

2017-04-15

To enhance survival and generation of definitive endoderm cells from human embryonic stem cells in a simple and reproducible system. Definitive endoderm (DE) differentiation from human embryonic stem cells (hESCs) was induced under a chemical-defined condition withdrawn insulin supplement and serum albumin. We dissected influence of "alternative growth factors", WNT3A, BMP4 and bFGF in activin A-driven differentiation by detection of DE-associated genes expression and cell viability. Expression of DE-associated SOX17 and FOXA2 genes was analyzed by real time reverse transcription polymerase chain reaction (RT-PCR) and Western blot assays. Quantitative evaluation of DE efficiency was performed by flow cytometry analysis of CXCR4-expressed cell population. Cell viability during DE differentiation was analyzed by an Annexin V/PI double staining test. Supplementation with WNT3A, BMP4 or bFGF promoted DE generation in a dose- and time-dependent manner. Cell apoptosis elicited by activin A was significantly ameliorated by a cocktail with WNT3A, BMP4 and bFGF. This allowed for sustained cell viability without insulin-containing supplements, thereby indirectly improving the efficiency of DE generation. Therefore, the cocktail containing is optimal for efficient DE generation in the presence of activin A and an insulin/albumin-free condition. This optimal condition facilitates the balance between the productivity and the viability maintenance, and could be valuable for mass production of DE with minimal variation. Copyright © 2017 Elsevier Inc. All rights reserved.

Evaluation of analytical performance and reliability of direct nanoLC-nanoESI-high resolution mass spectrometry for profiling the (xeno)metabolome.

PubMed

Chetwynd, Andrew J; David, Arthur; Hill, Elizabeth M; Abdul-Sada, Alaa

2014-10-01

Mass spectrometry (MS) profiling techniques are used for analysing metabolites and xenobiotics in biofluids; however, detection of low abundance compounds using conventional MS techniques is poor. To counter this, nanoflow ultra-high-pressure liquid chromatography-nanoelectrospray ionization-time-of-flight MS (nUHPLC-nESI-TOFMS), which has been used primarily for proteomics, offers an innovative prospect for profiling small molecules. Compared to conventional UHPLC-ESI-TOFMS, nUHPLC-nESI-TOFMS enhanced detection limits of a variety of (xeno)metabolites by between 2 and 2000-fold. In addition, this study demonstrates for the first time excellent repeatability and reproducibility for analysis of urine and plasma samples using nUHPLC-nESI-TOFMS, supporting implementation of this platform as a novel approach for high-throughput (xeno)metabolomics. Copyright © 2014 John Wiley & Sons, Ltd.

Circulating Organ-Specific MicroRNAs Serve as Biomarkers in Organ-Specific Diseases: Implications for Organ Allo- and Xeno-Transplantation

PubMed Central

Zhou, Ming; Hara, Hidetaka; Dai, Yifan; Mou, Lisha; Cooper, David K. C.; Wu, Changyou; Cai, Zhiming

2016-01-01

Different cell types possess different miRNA expression profiles, and cell/tissue/organ-specific miRNAs (or profiles) indicate different diseases. Circulating miRNA is either actively secreted by living cells or passively released during cell death. Circulating cell/tissue/organ-specific miRNA may serve as a non-invasive biomarker for allo- or xeno-transplantation to monitor organ survival and immune rejection. In this review, we summarize the proof of concept that circulating organ-specific miRNAs serve as non-invasive biomarkers for a wide spectrum of clinical organ-specific manifestations such as liver-related disease, heart-related disease, kidney-related disease, and lung-related disease. Furthermore, we summarize how circulating organ-specific miRNAs may have advantages over conventional methods for monitoring immune rejection in organ transplantation. Finally, we discuss the implications and challenges of applying miRNA to monitor organ survival and immune rejection in allo- or xeno-transplantation. PMID:27490531

Comparison of Synthetic Media Designed for Expansion of Adipose-Derived Mesenchymal Stromal Cells.

PubMed

Lensch, Michelle; Muise, Angela; White, Lisa; Badowski, Michael; Harris, David

2018-05-14

Mesenchymal stromal cells (MSCs) are multipotent cells that can differentiate into various cell types, such as osteoblasts, myocytes, and adipocytes. This characteristic makes the cells a useful tool in developing new therapies for a number of common maladies and diseases. The utilization of animal-derived growth serum, such as fetal bovine serum (FBS), for the expansion of MSCs has traditionally been used for cell culture. However, in clinical applications, animal-derived products present limitations and safety concerns for the recipient, as exposure to animal (xeno-) antigens and infectious agents is possible. Multiple synthetic, xeno-free media have been developed to combat these limitations of animal-derived growth serum and have the potential to be used in ex vivo MSC expansion for clinical use. The goal of this study was to determine if xeno-free media are adequate to significantly and efficiently expand MSCs derived from adipose tissue. MSCs were cultured in both standard FBS-containing as well as xeno-free media. The media were compared for cell yield, viability, and phenotypic expression via flow cytometry and directed differentiation. The xeno-free media that were tested were StemMACS MSC Expansion Media (Miltenyi Biotec, Bergisch Gladbach, Germany), PLTMax Human Platelet Lysate (Sigma-Aldrich, St. Louis, MO, USA), and MesenCult-hPL media (Stemcell Technologies, Vancouver, BC, Canada). All xeno-free media showed promise as a feasible replacement for animal-derived growth serums. The xeno-free media expanded MSCs more quickly than the FBS-containing medium and also showed great similarity in cell viability and phenotypic expression. In fact, each xeno-free media produced a greater viable cell yield than the standard FBS-containing medium.

A chemically defined substrate for the expansion and neuronal differentiation of human pluripotent stem cell-derived neural progenitor cells.

PubMed

Tsai, Yihuan; Cutts, Josh; Kimura, Azuma; Varun, Divya; Brafman, David A

2015-07-01

Due to the limitation of current pharmacological therapeutic strategies, stem cell therapies have emerged as a viable option for treating many incurable neurological disorders. Specifically, human pluripotent stem cell (hPSC)-derived neural progenitor cells (hNPCs), a multipotent cell population that is capable of near indefinite expansion and subsequent differentiation into the various cell types that comprise the central nervous system (CNS), could provide an unlimited source of cells for such cell-based therapies. However the clinical application of these cells will require (i) defined, xeno-free conditions for their expansion and neuronal differentiation and (ii) scalable culture systems that enable their expansion and neuronal differentiation in numbers sufficient for regenerative medicine and drug screening purposes. Current extracellular matrix protein (ECMP)-based substrates for the culture of hNPCs are expensive, difficult to isolate, subject to batch-to-batch variations, and, therefore, unsuitable for clinical application of hNPCs. Using a high-throughput array-based screening approach, we identified a synthetic polymer, poly(4-vinyl phenol) (P4VP), that supported the long-term proliferation and self-renewal of hNPCs. The hNPCs cultured on P4VP maintained their characteristic morphology, expressed high levels of markers of multipotency, and retained their ability to differentiate into neurons. Such chemically defined substrates will eliminate critical roadblocks for the utilization of hNPCs for human neural regenerative repair, disease modeling, and drug discovery. Copyright © 2015. Published by Elsevier B.V.

Defining Multiple Chronic Conditions for Quality Measurement.

PubMed

Drye, Elizabeth E; Altaf, Faseeha K; Lipska, Kasia J; Spatz, Erica S; Montague, Julia A; Bao, Haikun; Parzynski, Craig S; Ross, Joseph S; Bernheim, Susannah M; Krumholz, Harlan M; Lin, Zhenqiu

2018-02-01

Patients with multiple chronic conditions (MCCs) are a critical but undefined group for quality measurement. We present a generally applicable systematic approach to defining an MCC cohort of Medicare fee-for-service beneficiaries that we developed for a national quality measure, risk-standardized rates of unplanned admissions for Accountable Care Organizations. To define the MCC cohort we: (1) identified potential chronic conditions; (2) set criteria for cohort conditions based on MCC framework and measure concept; (3) applied the criteria informed by empirical analysis, experts, and the public; (4) described "broader" and "narrower" cohorts; and (5) selected final cohort with stakeholder input. Subjects were patients with chronic conditions. Participants included 21.8 million Medicare fee-for-service beneficiaries in 2012 aged 65 years and above with ≥1 of 27 Medicare Chronic Condition Warehouse condition(s). In total, 10 chronic conditions were identified based on our criteria; 8 of these 10 were associated with notably increased admission risk when co-occurring. A broader cohort (2+ of the 8 conditions) included 4.9 million beneficiaries (23% of total cohort) with an admission rate of 70 per 100 person-years. It captured 53% of total admissions. The narrower cohort (3+ conditions) had 2.2 million beneficiaries (10%) with 100 admissions per 100 person-years and captured 32% of admissions. Most stakeholders viewed the broader cohort as best aligned with the measure concept. By systematically narrowing chronic conditions to those most relevant to the outcome and incorporating stakeholder input, we defined an MCC admission measure cohort supported by stakeholders. This approach can be used as a model for other MCC outcome measures.

Fibroblast Growth Factors and Vascular Endothelial Growth Factor Promote Cardiac Reprogramming under Defined Conditions

PubMed Central

Yamakawa, Hiroyuki; Muraoka, Naoto; Miyamoto, Kazutaka; Sadahiro, Taketaro; Isomi, Mari; Haginiwa, Sho; Kojima, Hidenori; Umei, Tomohiko; Akiyama, Mizuha; Kuishi, Yuki; Kurokawa, Junko; Furukawa, Tetsushi; Fukuda, Keiichi; Ieda, Masaki

2015-01-01

Summary Fibroblasts can be directly reprogrammed into cardiomyocyte-like cells (iCMs) by overexpression of cardiac transcription factors, including Gata4, Mef2c, and Tbx5; however, this process is inefficient under serum-based culture conditions, in which conversion of partially reprogrammed cells into fully reprogrammed functional iCMs has been a major hurdle. Here, we report that a combination of fibroblast growth factor (FGF) 2, FGF10, and vascular endothelial growth factor (VEGF), termed FFV, promoted cardiac reprogramming under defined serum-free conditions, increasing spontaneously beating iCMs by 100-fold compared with those under conventional serum-based conditions. Mechanistically, FFV activated multiple cardiac transcriptional regulators and converted partially reprogrammed cells into functional iCMs through the p38 mitogen-activated protein kinase and phosphoinositol 3-kinase/AKT pathways. Moreover, FFV enabled cardiac reprogramming with only Mef2c and Tbx5 through the induction of cardiac reprogramming factors, including Gata4. Thus, defined culture conditions promoted the quality of cardiac reprogramming, and this finding provides new insight into the mechanism of cardiac reprogramming. PMID:26626177

Fibroblast Growth Factors and Vascular Endothelial Growth Factor Promote Cardiac Reprogramming under Defined Conditions.

PubMed

Yamakawa, Hiroyuki; Muraoka, Naoto; Miyamoto, Kazutaka; Sadahiro, Taketaro; Isomi, Mari; Haginiwa, Sho; Kojima, Hidenori; Umei, Tomohiko; Akiyama, Mizuha; Kuishi, Yuki; Kurokawa, Junko; Furukawa, Tetsushi; Fukuda, Keiichi; Ieda, Masaki

2015-12-08

Fibroblasts can be directly reprogrammed into cardiomyocyte-like cells (iCMs) by overexpression of cardiac transcription factors, including Gata4, Mef2c, and Tbx5; however, this process is inefficient under serum-based culture conditions, in which conversion of partially reprogrammed cells into fully reprogrammed functional iCMs has been a major hurdle. Here, we report that a combination of fibroblast growth factor (FGF) 2, FGF10, and vascular endothelial growth factor (VEGF), termed FFV, promoted cardiac reprogramming under defined serum-free conditions, increasing spontaneously beating iCMs by 100-fold compared with those under conventional serum-based conditions. Mechanistically, FFV activated multiple cardiac transcriptional regulators and converted partially reprogrammed cells into functional iCMs through the p38 mitogen-activated protein kinase and phosphoinositol 3-kinase/AKT pathways. Moreover, FFV enabled cardiac reprogramming with only Mef2c and Tbx5 through the induction of cardiac reprogramming factors, including Gata4. Thus, defined culture conditions promoted the quality of cardiac reprogramming, and this finding provides new insight into the mechanism of cardiac reprogramming. Copyright © 2015 The Authors. Published by Elsevier Inc. All rights reserved.

Clinical-Grade Manufacturing of Therapeutic Human Mesenchymal Stem/Stromal Cells in Microcarrier-Based Culture Systems.

PubMed

Fernandes-Platzgummer, Ana; Carmelo, Joana G; da Silva, Cláudia Lobato; Cabral, Joaquim M S

2016-01-01

The therapeutic potential of mesenchymal stem/stromal cells (MSC) has triggered the need for high cell doses in a vast number of clinical applications. This demand requires the development of good manufacturing practices (GMP)-compliant ex vivo expansion protocols that should be effective to deliver a robust and reproducible supply of clinical-grade cells in a safe and cost-effective manner. Controlled stirred-tank bioreactor systems under xenogeneic (xeno)-free culture conditions offer ideal settings to develop and optimize cell manufacturing to meet the standards and needs of human MSC for cellular therapies. Herein we describe two microcarrier-based stirred culture systems using spinner flasks and controlled stirred-tank bioreactors under xeno-free conditions for the efficient ex vivo expansion of human bone marrow and adipose tissue-derived MSC.

Improved Metabolic Health Alters Host Metabolism in Parallel with Changes in Systemic Xeno-Metabolites of Gut Origin

PubMed Central

Fiehn, Oliver; Chandler, Carol J.; Burnett, Dustin J.; Souza, Elaine C.; Casazza, Gretchen A.; Gustafson, Mary B.; Keim, Nancy L.; Newman, John W.; Hunter, Gary R.; Fernandez, Jose R.; Garvey, W. Timothy; Harper, Mary-Ellen; Hoppel, Charles L.; Meissen, John K.; Take, Kohei; Adams, Sean H.

2014-01-01

Novel plasma metabolite patterns reflective of improved metabolic health (insulin sensitivity, fitness, reduced body weight) were identified before and after a 14–17 wk weight loss and exercise intervention in sedentary, obese insulin-resistant women. To control for potential confounding effects of diet- or microbiome-derived molecules on the systemic metabolome, sampling was during a tightly-controlled feeding test week paradigm. Pairwise and multivariate analysis revealed intervention- and insulin-sensitivity associated: (1) Changes in plasma xeno-metabolites (“non-self” metabolites of dietary or gut microbial origin) following an oral glucose tolerance test (e.g. higher post-OGTT propane-1,2,3-tricarboxylate [tricarballylic acid]) or in the overnight-fasted state (e.g., lower γ-tocopherol); (2) Increased indices of saturated very long chain fatty acid elongation capacity; (3) Increased post-OGTT α-ketoglutaric acid (α-KG), fasting α-KG inversely correlated with Matsuda index, and altered patterns of malate, pyruvate and glutamine hypothesized to stem from improved mitochondrial efficiency and more robust oxidation of glucose. The results support a working model in which improved metabolic health modifies host metabolism in parallel with altering systemic exposure to xeno-metabolites. This highlights that interpretations regarding the origins of peripheral blood or urinary “signatures” of insulin resistance and metabolic health must consider the potentially important contribution of gut-derived metabolites toward the host's metabolome. PMID:24416208

Associations among the cecal microbiome and bacterially-derived cecal xeno-metabolites during diabetes progression in the UC Davis-Type 2 diabetes rat model

USDA-ARS?s Scientific Manuscript database

The gut microbiome is altered in obesity and diabetes, but the molecular signals linking gut microbes and host metabolic regulation have not been established. Our aim was to identify gut microbe-derived xeno-metabolites that associate with alterations in the microbiome during the progression of a t...

A novel efficient feeder-free culture system for the derivation of human induced pluripotent stem cells

PubMed Central

Nakagawa, Masato; Taniguchi, Yukimasa; Senda, Sho; Takizawa, Nanako; Ichisaka, Tomoko; Asano, Kanako; Morizane, Asuka; Doi, Daisuke; Takahashi, Jun; Nishizawa, Masatoshi; Yoshida, Yoshinori; Toyoda, Taro; Osafune, Kenji; Sekiguchi, Kiyotoshi; Yamanaka, Shinya

2014-01-01

In order to apply human embryonic stem cells (hESCs) and induced pluripotent stem cells (hiPSCs) to regenerative medicine, the cells should be produced under restricted conditions conforming to GMP guidelines. Since the conventional culture system has some issues that need to be addressed to achieve this goal, we developed a novel culture system. We found that recombinant laminin-511 E8 fragments are useful matrices for maintaining hESCs and hiPSCs when used in combination with a completely xeno-free (Xf) medium, StemFit™. Using this system, hESCs and hiPSCs can be easily and stably passaged by dissociating the cells into single cells for long periods, without any karyotype abnormalities. Human iPSCs could be generated under feeder-free (Ff) and Xf culture systems from human primary fibroblasts and blood cells, and they possessed differentiation abilities. These results indicate that hiPSCs can be generated and maintained under this novel Ff and Xf culture system. PMID:24399248

Estrogenic and androgenic activities of TBBA and TBMEPH, metabolites of novel brominated flame retardants, and selected bisphenols, using the XenoScreen XL YES/YAS assay.

PubMed

Fic, Anja; Žegura, Bojana; Gramec, Darja; Mašič, Lucija Peterlin

2014-10-01

The present study investigated and compared the estrogenic and androgenic activities of the three different classes of environmental pollutants and their metabolites using the XenoScreen XL YES/YAS assay, which has advantages compared with the original YES/YAS protocol. Contrary to the parent brominated flame retardants TBB and TBPH, which demonstrated no or very weak (anti)estrogenic or (anti)androgenic activities, their metabolites, TBBA and TBMEPH, exhibited anti-estrogenic (IC50 for TBBA=31.75 μM and IC50 for TBMEPH=0.265 μM) and anti-androgenic (IC50 for TBBA=73.95 μM and IC50 for TBMEPH=2.92 μM) activities. These results reveal that metabolism can enhance the anti-estrogenic and anti-androgenic effects of these two novel brominated flame retardants. Based on the activities of BPAF, BPF, BPA and MBP, we can conclude that the XenoScreen XL YES/YAS assay gives comparable results to the (anti)estrogenic or (anti)androgenic assays that are reported in the literature. For BPA, it was confirmed previously that the metabolite formed after an ipso-reaction (hydroxycumyl alcohol) exhibited higher estrogenic activity compared with the parent BPA, but this was not confirmed for BPAF and BPF ipso-metabolites, which were not active in the XenoScreen YES/YAS assay. Among the substituted BPA analogues, bis-GMA exhibited weak anti-estrogenic activity, BADGE demonstrated weak anti-estrogenic and anti-androgenic activities (IC50=13.73 μM), and the hydrolysed product BADGE·2H2O demonstrated no (anti)estrogenic or (anti)androgenic activities. Copyright © 2014. Published by Elsevier Ltd.

Defining the Role of Free Flaps in Partial Breast Reconstruction.

PubMed

Smith, Mark L; Molina, Bianca J; Dayan, Erez; Jablonka, Eric M; Okwali, Michelle; Kim, Julie N; Dayan, Joseph H

2018-03-01

 Free flaps have a well-established role in breast reconstruction after mastectomy; however, their role in partial breast reconstruction remains poorly defined. We reviewed our experience with partial breast reconstruction to better understand indications for free tissue transfer.  A retrospective review was performed of all patients undergoing partial breast reconstruction at our center between February 2009 and October 2015. We evaluated the characteristics of patients who underwent volume displacement procedures versus volume replacement procedures and free versus pedicled flap reconstruction.  There were 78 partial breast reconstructions, with 52 reductions/tissue rearrangements (displacement group) and 26 flaps (replacement group). Bra cup size and body mass index (BMI) were significantly smaller in the replacement group. Fifteen pedicled and 11 free flaps were performed. Most pedicled flaps (80.0%) were used for lateral or upper pole defects. Most free flaps (72.7%) were used for medial and inferior defects or when there was inadequate donor tissue for a pedicled flap. Complications included hematoma, cellulitis, and one aborted pedicled flap.  Free and pedicled flaps are useful for partial breast reconstruction, particularly in breast cancer patients with small breasts undergoing breast-conserving treatment (BCT). Flap selection depends on defect size, location, and donor tissue availability. Medial defects are difficult to reconstruct using pedicled flaps due to arc of rotation and intervening breast tissue. Free tissue transfer can overcome these obstacles. Confirming negative margins before flap reconstruction ensures harvest of adequate volume and avoids later re-operation. Judicious use of free flaps for oncoplastic reconstruction expands the possibility for breast conservation. Thieme Medical Publishers 333 Seventh Avenue, New York, NY 10001, USA.

Defined culture medium for stem cell differentiation: applicability of serum-free conditions in the mouse embryonic stem cell test.

PubMed

Riebeling, Christian; Schlechter, Katharina; Buesen, Roland; Spielmann, Horst; Luch, Andreas; Seiler, Andrea

2011-06-01

The embryonic stem cell test (EST) is a validated method to assess the developmental toxicity potency of chemicals. It was developed to reduce animal use and allow faster testing for hazard assessment. The cells used in this method are maintained and differentiated in media containing foetal calf serum. This animal product is of considerable variation in quality, and individual batches require extensive testing for their applicability in the EST. Moreover, its production involves a large number of foetuses and possible animal suffering. We demonstrate the serum-free medium and feeder cell-free maintenance of the mouse embryonic stem cell line D3 and investigate the use of specific growth factors for induction of cardiac differentiation. Using a combination of bone morphogenetic protein-2, bone morphogenetic protein-4, activin A and ascorbic acid, embryoid bodies efficiently differentiated into contracting myocardium. Additionally, examining levels of intracellular marker proteins by flow cytometry not only confirmed differentiation into cardiomyocytes, but demonstrated significant differentiation into neuronal cells in the same time frame. Thus, this approach might allow for simultaneous detection of developmental effects on both early mesodermal and neuroectodermal differentiation. The serum-free conditions for maintenance and differentiation of D3 cells described here enhance the transferability and standardisation and hence the performance of the EST. Copyright © 2011 Elsevier Ltd. All rights reserved.

Polymeric nanofibrous substrates stimulate pluripotent stem cells to form three-dimensional multilayered patty-like spheroids in feeder-free culture and maintain their pluripotency.

PubMed

Alamein, Mohammad A; Wolvetang, Ernst J; Ovchinnikov, Dmitry A; Stephens, Sebastien; Sanders, Katherine; Warnke, Patrick H

2015-09-01

Expansion of pluripotent stem cells in defined media devoid of animal-derived feeder cells to generate multilayered three-dimensional (3D) bulk preparations or spheroids, rather than two-dimensional (2D) monolayers, is advantageous for many regenerative, biological or disease-modelling studies. Here we show that electrospun polymer matrices comprised of nanofibres that mimic the architecture of the natural fibrous extracellular matrix allow for feeder-free expansion of pluripotent human induced pluripotent stem cells (IPSCs) and human embryonic stem cells (HESCs) into multilayered 3D 'patty-like' spheroid structures in defined xeno-free culture medium. The observation that IPSCs and HESCs readily revert to 2D growth in the absence of the synthetic nanofibre membranes suggests that this 3D expansion behaviour is mediated by the physical microenvironment and artificial niche provided by the nanofibres only. Importantly, we could show that such 3D growth as patties maintained the pluripotency of cells as long as they were kept on nanofibres. The generation of complex multilayered 3D structures consisting of only pluripotent cells on biodegradable nanofibre matrices of the desired shape and size will enable both industrial-scale expansion and intricate organ-tissue engineering applications with human pluripotent stem cells, where simultaneous coupling of differentiation pathways of all germ layers from one stem cell source may be required for organ formation. Copyright © 2014 John Wiley & Sons, Ltd.

A xenogeneic-free bioreactor system for the clinical-scale expansion of human mesenchymal stem/stromal cells.

PubMed

Dos Santos, Francisco; Campbell, Andrew; Fernandes-Platzgummer, Ana; Andrade, Pedro Z; Gimble, Jeffrey M; Wen, Yuan; Boucher, Shayne; Vemuri, Mohan C; da Silva, Cláudia L; Cabral, Joaquim M S

2014-06-01

The large cell doses (>1 × 10(6)  cells/kg) used in clinical trials with mesenchymal stem/stromal cells (MSC) will require an efficient production process. Moreover, monitoring and control of MSC ex-vivo expansion is critical to provide a safe and reliable cell product. Bioprocess engineering approaches, such as bioreactor technology, offer the adequate tools to develop and optimize a cost-effective culture system for the rapid expansion of human MSC for cellular therapy. Herein, a xenogeneic (xeno)-free microcarrier-based culture system was successfully established for bone marrow (BM) MSC and adipose tissue-derived stem/stromal cell (ASC) cultivation using a 1L-scale controlled stirred-tank bioreactor, allowing the production of (1.1 ± 0.1) × 10(8) and (4.5 ± 0.2) × 10(7) cells for BM MSC and ASC, respectively, after 7 days. Additionally, the effect of different percent air saturation values (%Airsat ) and feeding regime on the proliferation and metabolism of BM MSC was evaluated. No significant differences in cell growth and metabolic patterns were observed under 20% and 9%Airsat . Also, the three different feeding regimes studied-(i) 25% daily medium renewal, (ii) 25% medium renewal every 2 days, and (iii) fed-batch addition of concentrated nutrients and growth factors every 2 days-yielded similar cell numbers, and only slight metabolic differences were observed. Moreover, the immunophenotype (positive for CD73, CD90 and CD105 and negative for CD31, CD80 and HLA-DR) and multilineage differentiative potential of expanded cells were not affected upon bioreactor culture. These results demonstrated the feasibility of expanding human MSC from different sources in a clinically relevant expansion configuration in a controlled microcarrier-based stirred culture system under xeno-free conditions. The further optimization of this bioreactor culture system will represent a crucial step towards an efficient GMP-compliant clinical-scale MSC

Cross-sensitization between xeno- and allo-antigens on subsequent allogeneic and xenogeneic pancreatic islet transplantation in a murine model.

PubMed

Kim, Hyun-Je; Byun, Nari; Kwon, Ohsang; Park, Chung-Gyu

2016-11-18

The number of patients in need of organ transplantation is continuously on the rise. However, because of organ donor shortage, xenotransplantation has been highlighted as an alternative. Among the various porcine organs and tissues, porcine islets are considered to be the best-matching implantable candidates for clinical application based on recent progress in nonhuman primate pre-clinical studies. Nevertheless, before initiation of clinical trials, it should be confirmed whether the requisite xeno-antigen sensitization would have a deleterious effect on subsequent allo-transplantation or vice versa. Therefore, in the present study, the survival rate of islets grafted in naïve recipients was compared with that in cross-sensitized recipients. Enzyme-linked immunosorbent spot, fluorescence-activated cell sorting, and immunohistochemistry were conducted to assess the cellular and humoral immune responses. The survival days of Balb/c mouse islets transplanted into B6 mice that had been previously sensitized with porcine cells (i.e., xeno-sensitized) showed no significant difference from that of naïve B6 mice. Moreover, the survival days of porcine islets transplanted into allo-antigen (Balb/c)-sensitized B6 recipients was not significantly different from that in naïve B6 mice. Furthermore, our data provide the first demonstration that the cellular xenogeneic immune response (against porcine antigen) measured by an enzyme-linked immunosorbent spot assay is not cross-reactive to the allogeneic immune responses in a murine islet transplantation model. These results suggest that clinical application of islet xenotransplantation is not likely to have a deleterious effect on subsequent allogeneic islet transplantation. Copyright © 2016 Elsevier Inc. All rights reserved.

Cryopreservation of pluripotent stem cell aggregates in defined protein-free formulation.

PubMed

Sart, Sébastien; Ma, Teng; Li, Yan

2013-01-01

Cultivation of undifferentiated pluripotent stem cells (PSCs) as aggregates has emerged as an efficient culture configuration, enabling rapid and controlled large scale expansion. Aggregate-based PSC cryopreservation facilitates the integrated process of cell expansion and cryopreservation, but its feasibility has not been demonstrated. The goals of current study are to assess the suitability of cryopreserving intact mouse embryonic stem cell (mESC) aggregates and investigate the effects of aggregate size and the formulation of cryopreservation solution on mESC survival and recovery. The results demonstrated the size-dependent cell survival and recovery of intact aggregates. In particular, the generation of reactive oxygen species (ROS) and caspase activation were reduced for small aggregates (109 ± 55 μm) compared to medium (245 ± 77 μm) and large (365 ± 141 μm) ones, leading to the improved cell recovery. In addition, a defined protein-free formulation was tested and found to promote the aggregate survival, eliminating the cell exposure to animal serum. The cryopreserved aggregates also maintained the pluripotent markers and the differentiation capacity into three-germ layers after thawing. In summary, the cryopreservation of small PSC aggregates in a defined protein-free formulation was shown to be a suitable approach toward a fully integrated expansion and cryopreservation process at large scale. Copyright © 2012 American Institute of Chemical Engineers (AIChE).

The Xeno-glycomics database (XDB): a relational database of qualitative and quantitative pig glycome repertoire.

PubMed

Park, Hae-Min; Park, Ju-Hyeong; Kim, Yoon-Woo; Kim, Kyoung-Jin; Jeong, Hee-Jin; Jang, Kyoung-Soon; Kim, Byung-Gee; Kim, Yun-Gon

2013-11-15

In recent years, the improvement of mass spectrometry-based glycomics techniques (i.e. highly sensitive, quantitative and high-throughput analytical tools) has enabled us to obtain a large dataset of glycans. Here we present a database named Xeno-glycomics database (XDB) that contains cell- or tissue-specific pig glycomes analyzed with mass spectrometry-based techniques, including a comprehensive pig glycan information on chemical structures, mass values, types and relative quantities. It was designed as a user-friendly web-based interface that allows users to query the database according to pig tissue/cell types or glycan masses. This database will contribute in providing qualitative and quantitative information on glycomes characterized from various pig cells/organs in xenotransplantation and might eventually provide new targets in the α1,3-galactosyltransferase gene-knock out pigs era. The database can be accessed on the web at http://bioinformatics.snu.ac.kr/xdb.

Scalable Expansion of Human Pluripotent Stem Cell-Derived Neural Progenitors in Stirred Suspension Bioreactor Under Xeno-free Condition.

PubMed

Nemati, Shiva; Abbasalizadeh, Saeed; Baharvand, Hossein

2016-01-01

Recent advances in neural differentiation technology have paved the way to generate clinical grade neural progenitor populations from human pluripotent stem cells. These cells are an excellent source for the production of neural cell-based therapeutic products to treat incurable central nervous system disorders such as Parkinson's disease and spinal cord injuries. This progress can be complemented by the development of robust bioprocessing technologies for large scale expansion of clinical grade neural progenitors under GMP conditions for promising clinical use and drug discovery applications. Here, we describe a protocol for a robust, scalable expansion of human neural progenitor cells from pluripotent stem cells as 3D aggregates in a stirred suspension bioreactor. The use of this platform has resulted in easily expansion of neural progenitor cells for several passages with a fold increase of up to 4.2 over a period of 5 days compared to a maximum 1.5-2-fold increase in the adherent static culture over a 1 week period. In the bioreactor culture, these cells maintained self-renewal, karyotype stability, and cloning efficiency capabilities. This approach can be also used for human neural progenitor cells derived from other sources such as the human fetal brain.

Coupling the Gaussian Free Fields with Free and with Zero Boundary Conditions via Common Level Lines

NASA Astrophysics Data System (ADS)

Qian, Wei; Werner, Wendelin

2018-06-01

We point out a new simple way to couple the Gaussian Free Field (GFF) with free boundary conditions in a two-dimensional domain with the GFF with zero boundary conditions in the same domain: Starting from the latter, one just has to sample at random all the signs of the height gaps on its boundary-touching zero-level lines (these signs are alternating for the zero-boundary GFF) in order to obtain a free boundary GFF. Constructions and couplings of the free boundary GFF and its level lines via soups of reflected Brownian loops and their clusters are also discussed. Such considerations show for instance that in a domain with an axis of symmetry, if one looks at the overlay of a single usual Conformal Loop Ensemble CLE3 with its own symmetric image, one obtains the CLE4-type collection of level lines of a GFF with mixed zero/free boundary conditions in the half-domain.

Growth and differentiation of a murine interleukin-3-producing myelomonocytic leukemia cell line in a protein-free chemically defined medium.

PubMed

Kajigaya, Y; Ikuta, K; Sasaki, H; Matsuyama, S

1990-10-01

We established the continuous growth of WEHI-3B D+ cells in protein-free chemically defined F-12 medium by stepwise decreases in the concentration of fetal calf serum. This cell line, designated as WEHI-3B-Y1, has now been propagated in protein-free F-12 medium for 3 years. The population-doubling time of the cells in culture is about 24 hr. WEHI-3B-Y1 cells are immature undifferentiated cells which show positive staining for naphthol ASD chloroacetate esterase and alpha-naphthyl butyrate esterase and spontaneously exhibit a low level of differentiation to mature granulocytes and macrophages. Medium conditioned by WEHI-3B-Y1 cells stimulated the proliferation of an interleukin-3 (IL-3)-dependent FDCP-2 cell line. This conditioned medium was shown to have erythroid burst-promoting activity when assayed using normal murine bone marrow. The colony formation of WEHI-3B-Y1 cells in semi-solid agar culture was not stimulated by purified recombinant human granulocyte colony-stimulating factor (rhG-CSF). However, in the presence of human transferrin, rhG-CSF enhanced the number of colonies of WEHI-3B-Y1 cells but did not induce their differentiation. These results suggest that WEHI-3B-Y1 cells cultured in protein-free medium produced murine IL-3. In addition, human G-CSF enhanced the clonal growth but did not induce the differentiation of WEHI-3B-Y1 cells cultured in serum-free medium.

Orion Aerodynamics for Hypersonic Free Molecular to Continuum Conditions

NASA Technical Reports Server (NTRS)

Moss, James N.; Greene, Francis A.; Boyles, Katie A.

2006-01-01

Numerical simulations are performed for the Orion Crew Module, previously known as the Crew Exploration Vehicle (CEV) Command Module, to characterize its aerodynamics during the high altitude portion of its reentry into the Earth's atmosphere, that is, from free molecular to continuum hypersonic conditions. The focus is on flow conditions similar to those that the Orion Crew Module would experience during a return from the International Space Station. The bulk of the calculations are performed with two direct simulation Monte Carlo (DSMC) codes, and these data are anchored with results from both free molecular and Navier-Stokes calculations. Results for aerodynamic forces and moments are presented that demonstrate their sensitivity to rarefaction, that is, for free molecular to continuum conditions (Knudsen numbers of 111 to 0.0003). Also included are aerodynamic data as a function of angle of attack for different levels of rarefaction and results that demonstrate the aerodynamic sensitivity of the Orion CM to a range of reentry velocities (7.6 to 15 km/s).

Epigenetic stability, adaptability, and reversibility in human embryonic stem cells

PubMed Central

Tompkins, Joshua D.; Hall, Christine; Chen, Vincent Chang-yi; Li, Arthur Xuejun; Wu, Xiwei; Hsu, David; Couture, Larry A.; Riggs, Arthur D.

2012-01-01

The stability of human embryonic stem cells (hESCs) is of critical importance for both experimental and clinical applications. We find that as an initial response to altered culture conditions, hESCs change their transcription profile for hundreds of genes and their DNA methylation profiles for several genes outside the core pluripotency network. After adaption to conditions of feeder-free defined and/or xeno-free culture systems, expression and DNA methylation profiles are quite stable for additional passaging. However, upon reversion to the original feeder-based culture conditions, numerous transcription changes are not reversible. Similarly, although the majority of DNA methylation changes are reversible, highlighting the plasticity of DNA methylation, a few are persistent. Collectively, this indicates these cells harbor a memory of culture history. For culture-induced DNA methylation changes, we also note an intriguing correlation: hypomethylation of regions 500–2440 bp upstream of promoters correlates with decreased expression, opposite to that commonly seen at promoter-proximal regions. Lastly, changes in regulation of G-coupled protein receptor pathways provide a partial explanation for many of the unique transcriptional changes observed during hESC adaptation and reverse adaptation. PMID:22802633

Ex Vivo Expansion of Human Mesenchymal Stem Cells in Defined Serum-Free Media

PubMed Central

Jung, Sunghoon; Panchalingam, Krishna M.; Rosenberg, Lawrence; Behie, Leo A.

2012-01-01

Human mesenchymal stem cells (hMSCs) are presently being evaluated for their therapeutic potential in clinical studies to treat various diseases, disorders, and injuries. To date, early-phase studies have indicated that the use of both autologous and allogeneic hMSCs appear to be safe; however, efficacy has not been demonstrated in recent late-stage clinical trials. Optimized cell bioprocessing protocols may enhance the efficacy as well as safety of hMSC therapeutics. Classical media used for generating hMSCs are typically supplemented with ill-defined supplements such as fetal bovine serum (FBS) or human-sourced alternatives. Ideally, culture media are desired to have well-defined serum-free formulations that support the efficient production of hMSCs while maintaining their therapeutic and differentiation capacity. Towards this objective, we review here current cell culture media for hMSCs and discuss medium development strategies. PMID:22645619

Embryonic Stem Cells: Isolation, Characterization and Culture

NASA Astrophysics Data System (ADS)

Amit, Michal; Itskovitz-Eldor, Joseph

Embryonic stem cells are pluripotent cells isolated from the mammalian blastocyst. Traditionally, these cells have been derived and cultured with mouse embryonic fibroblast (MEF) supportive layers, which allow their continuous growth in an undifferentiated state. However, for any future industrial or clinical application hESCs should be cultured in reproducible, defined, and xeno-free culture system, where exposure to animal pathogens is prevented. From their derivation in 1998 the methods for culturing hESCs were significantly improved. This chapter wills discuss hESC characterization and the basic methods for their derivation and maintenance.

Defining And Employing Reference Conditions For Ecological Restoration Of The Lower Missouri River, USA

NASA Astrophysics Data System (ADS)

Jacobson, R. B.; Elliott, C. M.; Reuter, J. M.

2008-12-01

Ecological reference conditions are especially challenging for large, intensively managed rivers like the Lower Missouri. Historical information provides broad understanding of how the river has changed, but translating historical information into quantitative reference conditions remains a challenge. Historical information is less available for biological and chemical conditions than for physical conditions. For physical conditions, much of the early historical condition is documented in date-specific measurements or maps, and it is difficult to determine how representative these conditions are for a river system that was characterized historically by large floods and high channel migration rates. As an alternative to a historically defined least- disturbed condition, spatial variation within the Missouri River basin provides potential for defining a best- attainable reference condition. A possibility for the best-attainable condition for channel morphology is an unchannelized segment downstream of the lowermost dam (rkm 1298 - 1203). This segment retains multiple channels and abundant sandbars although it has a highly altered flow regime and a greatly diminished sediment supply. Conversely, downstream river segments have more natural flow regimes, but have been narrowed and simplified for navigation and bank stability. We use two computational tools to compensate for the lack of ideal reference conditions. The first is a hydrologic model that synthesizes natural and altered flow regimes based on 100 years of daily inputs to the river (daily routing model, DRM, US Army Corps of Engineers, 1998); the second tool is hydrodynamic modeling of habitat availability. The flow-regime and hydrodynamic outputs are integrated to define habitat-duration curves as the basis for reference conditions (least-disturbed flow regime and least-disturbed channel morphology). Lacking robust biological response models, we use mean residence time of water and a habitat diversity index as

Conditional Disease-Free Survival After Surgical Resection of Gastrointestinal Stromal Tumors

PubMed Central

Bischof, Danielle A.; Kim, Yuhree; Dodson, Rebecca; Jimenez, M. Carolina; Behman, Ramy; Cocieru, Andrei; Fisher, Sarah B.; Groeschl, Ryan T.; Squires, Malcolm H.; Maithel, Shishir K.; Blazer, Dan G.; Kooby, David A.; Gamblin, T. Clark; Bauer, Todd W.; Quereshy, Fayez A.; Karanicolas, Paul J.; Law, Calvin H. L.; Pawlik, Timothy M.

2015-01-01

IMPORTANCE Gastrointestinal stromal tumors (GISTs) are the most commonly diagnosed mesenchymal tumors of the gastrointestinal tract. The risk of recurrence following surgical resection of GISTs is typically reported from the date of surgery. However, disease-free survival (DFS) over time is dynamic and changes based on disease-free time already accumulated following surgery. OBJECTIVES To assess the comparative performance of established GIST recurrence risk prognostic scoring systems and to characterize conditional DFS following surgical resection of GISTs. DESIGN, SETTING, AND PARTICIPANTS A retrospective cohort study of 502 patients who underwent surgery for a primary, nonmetastatic GIST between January 1, 1998, and December 31, 2012, at 7 major academic cancer centers in the United States and Canada. MAIN OUTCOMES AND MEASURES Disease-free survival of the patients was classified according to 5 prognostic scoring systems, including the National Institutes of Health criteria, modified National Institutes of Health criteria, Memorial Sloan Kettering Cancer Center GIST nomogram, and American Joint Committee on Cancer gastric and nongastric categories. The concordance index (also known as the C statistic or the area under the receiver operating curve) of established GIST recurrence risk prognostic scoring systems. Conditional DFS estimates were calculated. RESULTS Overall 1-year, 3-year, and 5-year DFS following resection of GISTs was 95%, 83%, and 74%, respectively. All the prognostic scoring systems had fair prognostic ability. For all tumor sites, the American Joint Committee on Cancer gastric category demonstrated the best discrimination (C = 0.79). Using conditional DFS, the probability of remaining disease free for an additional 3 years given that a patient was disease free at 1 year, 3 years, and 5 years was 82%, 89%, and 92%, respectively. Patients with the highest initial recurrence risk demonstrated the greatest increase in conditional survival as time

Combined effect of smear layer characteristics and hydrostatic pulpal pressure on dentine bond strength of HEMA-free and HEMA-containing adhesives.

PubMed

Mahdan, Mohd Haidil Akmal; Nakajima, Masatoshi; Foxton, Richard M; Tagami, Junji

2013-10-01

This study evaluated the combined effect of smear layer characteristics with hydrostatic pulpal pressure (PP) on bond strength and nanoleakage expression of HEMA-free and -containing self-etch adhesives. Flat dentine surfaces were obtained from extracted human molars. Smear layers were created by grinding with #180- or #600-SiC paper. Three HEMA-free adhesives (Xeno V, G Bond Plus, Beautibond Multi) and two HEMA-containing adhesives (Bond Force, Tri-S Bond) were applied to the dentine surfaces under hydrostatic PP or none. Dentine bond strengths were determined using the microtensile bond test (μTBS). Data were statistically analyzed using three- and two-way ANOVA with Tukey post hoc comparison test. Nanoleakage evaluation was carried out under a scanning electron microscope (SEM). Coarse smear layer preparation and hydrostatic PP negatively affected the μTBS of HEMA-free and -containing adhesives, but there were no significant differences. The combined experimental condition significantly reduced μTBS of the HEMA-free adhesives, while the HEMA-containing adhesives exhibited no significant differences. Two-way ANOVA indicated that for HEMA-free adhesives, there were significant interactions in μTBS between smear layer characteristics and pulpal pressure, while for HEMA-containing adhesives, there were no significant interactions between them. Nanoleakage formation within the adhesive layers of both adhesive systems distinctly increased in the combined experimental group. The combined effect of coarse smear layer preparation with hydrostatic PP significantly reduced the μTBS of HEMA-free adhesives, while in HEMA-containing adhesives, these effects were not obvious. Smear layer characteristics and hydrostatic PP would additively compromise dentine bonding of self-etch adhesives, especially HEMA-free adhesives. Copyright © 2013 Elsevier Ltd. All rights reserved.

Canine and Equine Mesenchymal Stem Cells Grown in Serum Free Media Have Altered Immunophenotype.

PubMed

Clark, Kaitlin C; Kol, Amir; Shahbenderian, Salpi; Granick, Jennifer L; Walker, Naomi J; Borjesson, Dori L

2016-04-01

Mesenchymal stem cell (MSC) therapy is being increasingly used to treat dogs and horses with naturally-occurring diseases. However these animals also serve as critical large animal models for ongoing translation of cell therapy products to the human market. MSC manufacture for clinical use mandates improvement in cell culture systems to meet demands for higher MSC numbers and removal of xeno-proteins (i.e. fetal bovine serum, FBS). While serum-free media (SFM) is commercially available, its affects on MSC phenotype and immunomodulatory functions are not fully known. The objective of this study was to determine if specific MSC culture conditions, MSC expansion in HYPERFlasks® or MSC expansion in a commercially available SFM, would alter MSC proliferation, phenotype or immunomodulatory properties in vitro. MSCs cultured in HYPERFlasks® were similar in phenotype, proliferative capacity and immunomodulatory functions to MSCs grown in standard flasks however MSC yield was markedly increased. HYPERFlasks® therefore provide a viable option to generate greater cell numbers in a streamlined manner. Canine and equine MSCs expanded in SFM displayed similar proliferation, surface phenotype and inhibitory effect on lymphocyte proliferation in vitro. However, MSCs cultured in the absence of FBS secreted significantly less PGE2, and were significantly less able to inhibit IFNγ secretion by activated T-cells. Immunomodulatory functions altered by expansion in SFM were species dependent. Unlike equine MSCs, in canine adipose-derived MSCs, the inhibition of lymphocyte proliferation was not principally modulated by PGE2. The removal of FBS from both canine and equine MSC culture systems resulted in altered immunomodulatory properties in vitro and warrants further investigation prior to moving towards FBS-free culture conditions.

Lifesource XL-18 pedometer for measuring steps under controlled and free-living conditions.

PubMed

Liu, Sam; Brooks, Dina; Thomas, Scott; Eysenbach, Gunther; Nolan, Robert Peter

2015-01-01

The primary aim was to examine the criterion and construct validity and test-retest reliability of the Lifesource XL-18 pedometer (A&D Medical, Toronto, ON, Canada) for measuring steps under controlled and free-living activities. The influence of body mass index, waist size and walking speed on the criterion validity of XL-18 was also explored. Forty adults (35-74 years) performed a 6-min walk test in the controlled condition, and the criterion validity of XL-18 was assessed by comparing it to steps counted manually. Thirty-five adults participated in the free-living condition and the construct validity of XL-18 was assessed by comparing it to Yamax SW-200 (YAMAX Health & Sports, Inc., San Antonio, TX, USA). During the controlled condition, XL-18 did not significantly differ from criterion (P > 0.05) and no systematic error was found using Bland-Altman analysis. The accuracy of XL-18 decreased with slower walking speed (P = 0.001). During the free-living condition, Bland-Altman analysis revealed that XL-18 overestimated daily steps by 327 ± 118 than Yamax (P = 0.004). However, the absolute percent error (APE) (6.5 ± 0.58%) was still within an acceptable range. XL-18 did not differ statistically between pant pockets. XL-18 is suitable for measuring steps in controlled and free-living conditions. However, caution may be required when interpreting the steps recorded under slower speeds and free-living conditions.

Conditional Versus Unconditional Procedures for Sample-Free Item Analysis

ERIC Educational Resources Information Center

Wright, Benjamin D.; Douglas, Graham A.

1977-01-01

Two procedures for Rasch, sample-free item calibration are reviewed and compared for accuracy. The theoretically ideal "conditional" procedure is impractical for more than fifteen items. The more practical but biased "unconditional" procedure is discussed in detail. (Author/JKS)

Generation and maturation of bone marrow-derived DCs under serum-free conditions.

PubMed

Kim, Sung Jung; Diamond, Betty

2007-06-30

Standard protocols for the generation of murine dendritic cells (DCs) employ medium supplemented with heat-inactivated fetal calf serum (FCS). Recently, several attempts have been made to avoid serum exposure during DC culture. The impetus for these efforts has been a desire to generate DCs for clinical use, as preclinical data have demonstrated their efficacy in immune activation and in immune suppression both in vitro and in vivo. However, these protocols have resulted in contradictory outcomes with respect to DC survival in culture and activation status. In this report, we compared several serum-free culture conditions with respect to survival, differentiation, activation, and cytokine profile of murine DC progenitors. DC progenitors can survive only in some serum-free conditions. Surprisingly, DCs grown in serum-free medium display a higher expression of activation markers upon stimulation. They produce increased IL-12 and decreased IL-6 following stimulation. Furthermore, DCs derived under serum-free conditions may express unusual surface markers, B220 and Ly6C/G, implying an increased differentiation to plasmacytoid DCs (pDCs).

Keratinocytes propagated in serum-free, feeder-free culture conditions fail to form stratified epidermis in a reconstituted skin model.

PubMed

Lamb, Rebecca; Ambler, Carrie A

2013-01-01

Primary human epidermal stem cells isolated from skin tissues and subsequently expanded in tissue culture are used for human therapeutic use to reconstitute skin on patients and to generate artificial skin in culture for academic and commercial research. Classically, epidermal cells, known as keratinocytes, required fibroblast feeder support and serum-containing media for serial propagation. In alignment with global efforts to remove potential animal contaminants, many serum-free, feeder-free culture methods have been developed that support derivation and growth of these cells in 2-dimensional culture. Here we show that keratinocytes grown continually in serum-free and feeder-free conditions were unable to form into a stratified, mature epidermis in a skin equivalent model. This is not due to loss of cell potential as keratinocytes propagated in serum-free, feeder-free conditions retain their ability to form stratified epidermis when re-introduced to classic serum-containing media. Extracellular calcium supplementation failed to improve epidermis development. In contrast, the addition of serum to commercial, growth media developed for serum-free expansion of keratinocytes facilitated 3-dimensional stratification in our skin equivalent model. Moreover, the addition of heat-inactivated serum improved the epidermis structure and thickness, suggesting that serum contains factors that both aid and inhibit stratification.

Keratinocytes Propagated in Serum-Free, Feeder-Free Culture Conditions Fail to Form Stratified Epidermis in a Reconstituted Skin Model

PubMed Central

Lamb, Rebecca; Ambler, Carrie A.

2013-01-01

Primary human epidermal stem cells isolated from skin tissues and subsequently expanded in tissue culture are used for human therapeutic use to reconstitute skin on patients and to generate artificial skin in culture for academic and commercial research. Classically, epidermal cells, known as keratinocytes, required fibroblast feeder support and serum-containing media for serial propagation. In alignment with global efforts to remove potential animal contaminants, many serum-free, feeder-free culture methods have been developed that support derivation and growth of these cells in 2-dimensional culture. Here we show that keratinocytes grown continually in serum-free and feeder-free conditions were unable to form into a stratified, mature epidermis in a skin equivalent model. This is not due to loss of cell potential as keratinocytes propagated in serum-free, feeder-free conditions retain their ability to form stratified epidermis when re-introduced to classic serum-containing media. Extracellular calcium supplementation failed to improve epidermis development. In contrast, the addition of serum to commercial, growth media developed for serum-free expansion of keratinocytes facilitated 3-dimensional stratification in our skin equivalent model. Moreover, the addition of heat-inactivated serum improved the epidermis structure and thickness, suggesting that serum contains factors that both aid and inhibit stratification. PMID:23326335

Nonempirical Semilocal Free-Energy Density Functional for Matter under Extreme Conditions

DOE PAGES

Karasiev, Valentin V.; Dufty, James W.; Trickey, S. B.

2018-02-14

The potential for density functional calculations to predict the properties of matter under extreme conditions depends crucially upon having a non-empirical approximate free energy functional valid over a wide range of state conditions. Unlike the ground-state case, no such free-energy exchange- correlation (XC) functional exists. We remedy that with systematic construction of a generalized gradient approximation XC free-energy functional based on rigorous constraints, including the free energy gradient expansion. The new functional provides the correct temperature dependence in the slowly varying regime and the correct zero-T, high-T, and homogeneous electron gas limits. Application in Kohn-Sham calculations for hot electrons inmore » a static fcc Aluminum lattice demon- strates the combined magnitude of thermal and gradient effects handled by this functional. Its accuracy in the increasingly important warm dense matter regime is attested by excellent agreement of the calculated deuterium equation of state with reference path integral Monte Carlo results at intermediate and elevated temperatures and by low density Al calculations over a wide T range.« less

Nonempirical Semilocal Free-Energy Density Functional for Matter under Extreme Conditions

DOE Office of Scientific and Technical Information (OSTI.GOV)

Karasiev, Valentin V.; Dufty, James W.; Trickey, S. B.

The potential for density functional calculations to predict the properties of matter under extreme conditions depends crucially upon having a non-empirical approximate free energy functional valid over a wide range of state conditions. Unlike the ground-state case, no such free-energy exchange- correlation (XC) functional exists. We remedy that with systematic construction of a generalized gradient approximation XC free-energy functional based on rigorous constraints, including the free energy gradient expansion. The new functional provides the correct temperature dependence in the slowly varying regime and the correct zero-T, high-T, and homogeneous electron gas limits. Application in Kohn-Sham calculations for hot electrons inmore » a static fcc Aluminum lattice demon- strates the combined magnitude of thermal and gradient effects handled by this functional. Its accuracy in the increasingly important warm dense matter regime is attested by excellent agreement of the calculated deuterium equation of state with reference path integral Monte Carlo results at intermediate and elevated temperatures and by low density Al calculations over a wide T range.« less

Human fibroblast-derived extracellular matrix constructs for bone tissue engineering applications.

PubMed

Tour, Gregory; Wendel, Mikael; Tcacencu, Ion

2013-10-01

We exploited the biomimetic approach to generate constructs composed of synthetic biphasic calcium phosphate ceramic and extracellular matrix (SBC-ECM) derived from adult human dermal fibroblasts in complete xeno-free culture conditions. The construct morphology and composition were assessed by scanning electron microscopy, histology, immunohistochemistry, Western blot, glycosaminoglycan, and hydroxyproline assays. Residual DNA quantification, endotoxin testing, and local inflammatory response after implantation in a rat critical-sized calvarial defect were used to access the construct biocompatibility. Moreover, in vitro interaction of human mesenchymal stem cells (hMSCs) with the constructs was studied. The bone marrow- and adipose tissue-derived mesenchymal stem cells were characterized by flow cytometry and tested for osteogenic differentiation capacity prior seeding onto SBC-ECM, followed by alkaline phosphatase, 3-(4,5-dimethythiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay, and real-time quantitative polymerase chain reaction to assess the osteogenic differentiation of hMSCs after seeding onto the constructs at different time intervals. The SBC-ECM constructs enhanced osteogenic differentiation of hMSCs in vitro and exhibited excellent handling properties and high biocompatibility in vivo. Our results highlight the ability to generate in vitro fibroblast-derived ECM constructs in complete xeno-free conditions as a step toward clinical translation, and the potential use of SBC-ECM in craniofacial bone tissue engineering applications. Copyright © 2013 Wiley Periodicals, Inc., a Wiley Company.

Generation of hematopoietic stem cells from human embryonic stem cells using a defined, stepwise, serum-free, and serum replacement-free monolayer culture method.

PubMed

Kim, So-Jung; Jung, Ji-Won; Ha, Hye-Yeong; Koo, Soo Kyung; Kim, Eung-Gook; Kim, Jung-Hyun

2017-03-01

Embryonic stem cells (ESCs) can be expanded infinitely in vitro and have the potential to differentiate into hematopoietic stem cells (HSCs); thus, they are considered a useful source of cells for HSC production. Although several technical in vitro methods for engineering HSCs from pluripotent stem cells have been developed, clinical application of HSCs engineered from pluripotent stem cells is restricted because of the possibility of xenogeneic contamination resulting from the use of murine materials. Human ESCs (CHA-hES15) were cultured on growth factor-reduced Matrigel-coated dishes in the mTeSR1 serum-free medium. When the cells were 70% confluent, we initiated HSC differentiation by three methods involving (1) knockout serum replacement (KSR), cytokines, TGFb1, EPO, and FLT3L; (2) KSR, cytokines, and bFGF; or (3) cytokines and bFGF. Among the three differentiation methods, the minimal number of cytokines without KSR resulted in the greatest production of HSCs. The optimized method resulted in a higher proportion of CD34 + CD43 + hematopoietic progenitor cells (HPCs) and CD34 + CD45 + HPCs compared to the other methods. In addition, the HSCs showed the potential to differentiate into multiple lineages of hematopoietic cells in vitro . In this study, we optimized a two-step, serum-free, animal protein-free, KSR-free, feeder-free, chemically defined monolayer culture method for generation of HSCs and hematopoietic stem and progenitor cells (HSPCs) from human ESCs.

Efficient, Long Term Production of Monocyte-Derived Macrophages from Human Pluripotent Stem Cells under Partly-Defined and Fully-Defined Conditions

PubMed Central

van Wilgenburg, Bonnie; Browne, Cathy; Vowles, Jane; Cowley, Sally A.

2013-01-01

Human macrophages are specialised hosts for HIV-1, dengue virus, Leishmania and Mycobacterium tuberculosis. Yet macrophage research is hampered by lack of appropriate cell models for modelling infection by these human pathogens, because available myeloid cell lines are, by definition, not terminally differentiated like tissue macrophages. We describe here a method for deriving monocytes and macrophages from human Pluripotent Stem Cells which improves on previously published protocols in that it uses entirely defined, feeder- and serum-free culture conditions and produces very consistent, pure, high yields across both human Embryonic Stem Cell (hESC) and multiple human induced Pluripotent Stem Cell (hiPSC) lines over time periods of up to one year. Cumulatively, up to ∼3×107 monocytes can be harvested per 6-well plate. The monocytes produced are most closely similar to the major blood monocyte (CD14+, CD16low, CD163+). Differentiation with M-CSF produces macrophages that are highly phagocytic, HIV-1-infectable, and upon activation produce a pro-inflammatory cytokine profile similar to blood monocyte-derived macrophages. Macrophages are notoriously hard to genetically manipulate, as they recognise foreign nucleic acids; the lentivector system described here overcomes this, as pluripotent stem cells can be relatively simply genetically manipulated for efficient transgene expression in the differentiated cells, surmounting issues of transgene silencing. Overall, the method we describe here is an efficient, effective, scalable system for the reproducible production and genetic modification of human macrophages, facilitating the interrogation of human macrophage biology. PMID:23951090

Why Australia needs to define obesity as a chronic condition.

PubMed

Opie, C A; Haines, H M; Ervin, K E; Glenister, K; Pierce, D

2017-05-23

In Australia people with a diagnosed chronic condition can be managed on unique funded care plans that allow the recruitment of a multidisciplinary team to assist in setting treatment goals and adequate follow up. In contrast to the World Health Organisation, the North American and European Medical Associations, the Australian Medical Association does not recognise obesity as a chronic condition, therefore excluding a diagnosis of obesity from qualifying for a structured and funded treatment plan. BODY: The Australian guidelines for management of Obesity in adults in Primary Care are structured around a five step process -the '5As': Ask & Assess, Advise, Assist and Arrange'. This article aims to identify the key challenges and successes associated with the '5As' approach, to better understand the reasons for the gap between the high Australian prevalence of overweight and obesity and an actual diagnosis and treatment plan for managing obesity. It argues that until the Australian health system follows the international lead and defines obesity as a chronic condition, the capacity for Australian doctors to diagnose and initiate structured treatment plans will remain limited and ineffective. Australian General Practitioners are limited in their ability manage obesity, as the current treatment guidelines only recognise obesity as a risk factor rather than a chronic condition.

Nonempirical Semilocal Free-Energy Density Functional for Matter under Extreme Conditions.

PubMed

Karasiev, Valentin V; Dufty, James W; Trickey, S B

2018-02-16

Realizing the potential for predictive density functional calculations of matter under extreme conditions depends crucially upon having an exchange-correlation (XC) free-energy functional accurate over a wide range of state conditions. Unlike the ground-state case, no such functional exists. We remedy that with systematic construction of a generalized gradient approximation XC free-energy functional based on rigorous constraints, including the free-energy gradient expansion. The new functional provides the correct temperature dependence in the slowly varying regime and the correct zero-T, high-T, and homogeneous electron gas limits. Its accuracy in the warm dense matter regime is attested by excellent agreement of the calculated deuterium equation of state with reference path integral Monte Carlo results at intermediate and elevated T. Pressure shifts for hot electrons in compressed static fcc Al and for low-density Al demonstrate the combined magnitude of thermal and gradient effects handled well by this functional over a wide T range.

Nonempirical Semilocal Free-Energy Density Functional for Matter under Extreme Conditions

NASA Astrophysics Data System (ADS)

Karasiev, Valentin V.; Dufty, James W.; Trickey, S. B.

2018-02-01

Realizing the potential for predictive density functional calculations of matter under extreme conditions depends crucially upon having an exchange-correlation (X C ) free-energy functional accurate over a wide range of state conditions. Unlike the ground-state case, no such functional exists. We remedy that with systematic construction of a generalized gradient approximation X C free-energy functional based on rigorous constraints, including the free-energy gradient expansion. The new functional provides the correct temperature dependence in the slowly varying regime and the correct zero-T , high-T , and homogeneous electron gas limits. Its accuracy in the warm dense matter regime is attested by excellent agreement of the calculated deuterium equation of state with reference path integral Monte Carlo results at intermediate and elevated T . Pressure shifts for hot electrons in compressed static fcc Al and for low-density Al demonstrate the combined magnitude of thermal and gradient effects handled well by this functional over a wide T range.

Interaction of Strong Turbulence With Free Surfaces

NASA Astrophysics Data System (ADS)

Dalrymple, Robert A.

Spray from a nozzle, spilling breakers, and “rooster tails” from speeding boats are all examples of a turbulent flow with a free surface. In many cases like these, the free surface is difficult to discern as the volume of air in the fluid can exceed that of the water.In traditional studies, the free surface is simply defined as a continuous surface separating the fluid from air. The pressure at the surface is assumed to be atmospheric pressure and the fluid comprising the surface moves with the surface. While these conditions are sufficient for non-turbulent flows, such as nonbreaking water waves, and lead to the (albeit non-linear) dynamic and kinematic free surface boundary conditions that serve to provide sufficient conditions to determine the surface, they are not valid descriptions for a bubbly free surface in a highly turbulent regime, such as the roller in front of a spilling breaker or the propeller wash behind a ship.

Defining biophysical reference conditions for dynamics river systems: an Alaskan example

NASA Astrophysics Data System (ADS)

Pess, G. R.

2008-12-01

Defining reference conditions for dynamic river ecosystems is difficult for two reasons. First long-term, persistent anthropogenic influences such as land development, harvest of biological resources, and invasive species have resulted in degraded, reduced, and simplified ecological communities and associated habitats. Second, river systems that have not been altered through human disturbance rarely have a long-term dataset on ecological conditions. However there are exceptions which can help us define the dynamic nature of river ecosystems. One large-scale exception is the Wood River system in Bristol Bay, Alaska, where habitat and salmon populations have not been altered by anthropogenic influences such as land development, hatchery production, and invasive species. In addition, the one major anthropogenic disturbance, salmon (Oncorhynchus spp.) harvest, has been quantified and regulated since its inception. First, we examined the variation in watershed and stream habitat characteristics across the Wood River system. We then compared these stream habitat characteristics with data that was collected in the 1950s. Lastly, we examined the correlation between pink (Oncorhynchus gorbuscha), chum (O. keta), and Chinook (O. tshawytscha), and sockeye salmon (O. nerka), and habitat characteristics in the Wood River system using four decades of data on salmon. We found that specific habitat attributes such as stream channel wetted width, depth, cover type, and the proportion of spawnable area were similar to data collected in the 1950s. Greater stream habitat variation occurred among streams than over time. Salmon occurrence and abundance, however was more temporal and spatially variable. The occurrence of pink and chum salmon increased from the 1970's to the present in the Wood River system, while sockeye abundance has fluctuated with changes in ocean conditions. Pink, Chinook and chum salmon ranged from non-existent to episodic to abundantly perennial, while sockeye

Experimental verification of free-space singular boundary conditions in an invisibility cloak

NASA Astrophysics Data System (ADS)

Wu, Qiannan; Gao, Fei; Song, Zhengyong; Lin, Xiao; Zhang, Youming; Chen, Huanyang; Zhang, Baile

2016-04-01

A major issue in invisibility cloaking, which caused intense mathematical discussions in the past few years but still remains physically elusive, is the plausible singular boundary conditions associated with the singular metamaterials at the inner boundary of an invisibility cloak. The perfect cloaking phenomenon, as originally proposed by Pendry et al for electromagnetic waves, cannot be treated as physical before a realistic inner boundary of a cloak is demonstrated. Although a recent demonstration has been done in a waveguide environment, the exotic singular boundary conditions should apply to a general environment as in free space. Here we fabricate a metamaterial surface that exhibits the singular boundary conditions and demonstrate its performance in free space. Particularly, the phase information of waves reflected from this metamaterial surface is explicitly measured, confirming the singular responses of boundary conditions for an invisibility cloak.

Using Perceptual Signatures to Define and Dissociate Condition-Specific Neural Etiology: Autism and Fragile X Syndrome as Model Conditions

ERIC Educational Resources Information Center

Bertone, Armando; Hanck, Julie; Kogan, Cary; Chaudhuri, Avi; Cornish, Kim

2010-01-01

The functional link between genetic alteration and behavioral end-state is rarely straightforward and never linear. Cases where neurodevlopmental conditions defined by a distinct genetic etiology share behavioral phenotypes are exemplary, as is the case for autism and Fragile X Syndrome (FXS). In this paper and its companion paper, we propose a…

Stability studies needed to define the handling and transport conditions of sensitive pharmaceutical or biotechnological products.

PubMed

Ammann, Claude

2011-12-01

Many pharmaceutical or biotechnological products require transport using temperature-controlled systems to keep their therapeutic properties. There are presently no official guidelines for testing pharmaceutical products in order to define suitable transport specifications. After reviewing the current guidance documents, this paper proposes a methodology for testing pharmaceutical products and defining appropriate transport conditions.

Effects of conditioned reinforcement frequency in an intermittent free-feeding situation, 12

PubMed Central

Zimmerman, Joseph; Hanford, Peter V.; Brown, Wyman

1967-01-01

Key-pecking intermittently produced a set of brief exteroceptive stimulus changes under two-component multiple schedules of conditioned reinforcement. Throughout the study, free access to grain was concurrently provided on an intermittent basis via a variable-interval tape. Free food presentations scheduled by the tape were delivered if no peck had been emitted for 6 sec, and the brief stimulus changes produced by responding under the multiple schedules were those which accompanied food presentation. The second component of each multiple schedule was always associated with a 1-min, variable-interval schedule of conditioned reinforcement. The schedule associated with the first component was systematically varied and conditioned reinforcement was either absent (extinction) or programmed on a 1-, 3-, 6-, or 12-min variable-interval schedule. Under these conditions, rate of responding in the manipulated component decreased monotonically with a decrease in the frequency of conditioned reinforcement. In addition, contrast effects were often obtained in the constant, second component. These results are similar to those obtained with similar multiple schedules of primary reinforcement. PMID:6033554

Prefrontal Contribution to Decision-Making under Free-Choice Conditions

PubMed Central

Funahashi, Shintaro

2017-01-01

Executive function is thought to be the coordinated operation of multiple neural processes and allows to accomplish a current goal flexibly. The most important function of the prefrontal cortex is the executive function. Among a variety of executive functions in which the prefrontal cortex participates, decision-making is one of the most important. Although the prefrontal contribution to decision-making has been examined using a variety of behavioral tasks, recent studies using fMRI have shown that the prefrontal cortex participates in decision-making under free-choice conditions. Since decision-making under free-choice conditions represents the very first stage for any kind of decision-making process, it is important that we understand its neural mechanism. Although few studies have examined this issue while a monkey performed a free-choice task, those studies showed that, when the monkey made a decision to subsequently choose one particular option, prefrontal neurons showing selectivity to that option exhibited transient activation just before presentation of the imperative cue. Further studies have suggested that this transient increase is caused by the irregular fluctuation of spontaneous firing just before cue presentation, which enhances the response to the cue and biases the strength of the neuron's selectivity to the option. In addition, this biasing effect was observed only in neurons that exhibited sustained delay-period activity, indicating that this biasing effect not only influences the animal's decision for an upcoming choice, but also is linked to working memory mechanisms in the prefrontal cortex. PMID:28798662

Establishment of a Polychlorinated Biphenyl-Dechlorinating Microbial Consortium, Specific for Doubly Flanked Chlorines, in a Defined, Sediment-Free Medium

PubMed Central

Wu, Qingzhong; Sowers, Kevin R.; May, Harold D.

2000-01-01

Estuarine sediment from Charleston Harbor, South Carolina, was used as inoculum for the development of an anaerobic enrichment culture that specifically dechlorinates doubly flanked chlorines (i.e., chlorines bound to carbon that are flanked on both sides by other chlorine-carbon bonds) of polychlorinated biphenyls (PCBs). Dechlorination was restricted to the para chlorine in cultures enriched with 10 mM fumarate, 50 ppm (173 μM) 2,3,4,5-tetrachlorobiphenyl, and no sediment. Initially the rate of dechlorination decreased upon the removal of sediment from the medium. However, the dechlorinating activity was sustainable, and following sequential transfer in a defined, sediment-free estuarine medium, the activity increased to levels near that observed with sediment. The culture was nonmethanogenic, and molybdate, ampicillin, chloramphenicol, neomycin, and streptomycin inhibited dechlorination activity; bromoethanesulfonate and vancomycin did not. Addition of 17 PCB congeners indicated that the culture specifically removes double flanked chlorines, preferably in the para position, and does not attack ortho chlorines. This is the first microbial consortium shown to para or meta dechlorinate a PCB congener in a defined sediment-free medium. It is the second PCB-dechlorinating enrichment culture to be sustained in the absence of sediment, but its dechlorinating capabilities are entirely different from those of the other sediment-free PCB-dechlorinating culture, an ortho-dechlorinating consortium, and do not match any previously published Aroclor-dechlorinating patterns. PMID:10618202

Simulation of VSPT Experimental Cascade Under High and Low Free-Stream Turbulence Conditions

NASA Technical Reports Server (NTRS)

Ameri, Ali A.; Giel, Paul W.; Flegel, Ashlie

2014-01-01

Variable-Speed Power Turbines (VSPT) for rotorcraft applications operate in conditions of low Reynolds number and a wide range in incidence resulting from rotational speed variation. A comprehensive data set obtained in a linear cascade which includes the effects of Reynolds number, free-stream turbulence and incidence is now available and this paper concerns itself with the post-diction of boundary layer transitionseparation, blade pressure loading and total pressure loss pertaining to the conditions set for measurements in that data set. The distinction between the state of the measured data presented here and the earlier publications is the addition of high free-stream turbulence intensity. We will, for the purposes of the numerical post-diction, present some of the higher free stream turbulence data in this paper but defer a comprehensive presentation and treatment of the measured data will be done elsewhere.

Unraveling fabrication and calibration of wearable gas monitor for use under free-living conditions.

PubMed

Yue Deng; Cheng Chen; Tsow, Francis; Xiaojun Xian; Forzani, Erica

2016-08-01

Volatile organic compounds (VOC) are organic chemicals that have high vapor pressure at regular conditions. Some VOC could be dangerous to human health, therefore it is important to determine real-time indoor and outdoor personal exposures to VOC. To achieve this goal, our group has developed a wearable gas monitor with a complete sensor fabrication and calibration protocol for free-living conditions. Correction factors for calibrating the sensors, including sensitivity, aging effect, and temperature effect are implemented into a Quick Response Code (QR code), so that the pre-calibrated quartz tuning fork (QTF) sensor can be used with the wearable monitor under free-living conditions.

Influence of free-stall flooring on comfort and hygiene of dairy cows during warm climatic conditions.

PubMed

De Palo, P; Tateo, A; Zezza, F; Corrente, M; Centoducati, P

2006-12-01

An evaluation of behavioral and hygienic conditions was carried out with 4 materials used as free-stall flooring for dairy cows: polyethylene vinyl acetate (EVA) and polypropylene vinyl acetate (PVA) mats, wood shavings, and solid manure. The free-stall type selected by cows was evaluated in response to changes in environmental temperature and humidity. Two tests were used: 1) a preference test, in which 8 cows were housed in a pen with 32 free stalls and 4 types of flooring; and 2) an aversion test, in which 32 cows were placed in 4 pens, each with 8 free stalls. The free stalls in each pen had a single type of bedding material. These tests showed that the comfort of dairy cows was predominantly influenced by environmental conditions. The preference test for lying showed that cows preferred free-stall floors with EVA mats over those with PVA mats, wood shavings, and solid manure (332.4 +/- 24.0 vs. 130.8 +/- 6.2, 160.9 +/- 23.7, and 102.6 +/- 23.2 min/d, respectively), but under conditions of heat stress, with a temperature-humidity index > 80, they chose wood shavings and solid manure lying areas. These results were confirmed by the aversion test. In all experimental and environmental conditions, the PVA mats were the least suitable. The mats contaminated with organic manure and the free stalls bedded with wood shavings and organic solids did not differ in either the coliform load on the lying surfaces (EVA mats: 290 +/- 25; PVA mats: 306 +/- 33; wood shavings: 290 +/- 39; and solid manure: 305 +/- 23 log(10) cfu/mL) or the total bacterial count in the raw milk (EVA mats: 232 +/- 22; PVA mats: 233 + 24; wood shavings: 221 +/- 24; and solid manure: 220 +/- 25 log(10) cfu/mL). These results demonstrate that the comfort of dairy cows housed in barns with free stalls as resting areas does not depend only on the material used, but also on the value of the material in microenvironmental conditions.

Concurrent Validity of Wearable Activity Trackers Under Free-Living Conditions.

PubMed

Brooke, Skyler M; An, Hyun-Sung; Kang, Seoung-Ki; Noble, John M; Berg, Kris E; Lee, Jung-Min

2017-04-01

Brooke, SM, An, H-S, Kang, S-K, Noble, JM, Berg, KE, and Lee, J-M. Concurrent validity of wearable activity trackers under free-living conditions. J Strength Cond Res 31(4): 1097-1106, 2017-The purpose of this study is to evaluate the concurrent validity of wearable activity trackers in energy expenditure (EE) and sleep period time (SPT) under free-living conditions. Ninety-five (28.5 ± 9.8 years) healthy men (n = 34) and women (n = 61) participated in this study. The total EE and SPT were measured using 8 monitors: Nike+ FuelBand SE (NFB), Garmin VivoFit (VF), Misfit Shine (MF), Fitbit Flex (FF), Jawbone UP (JU), Polar Loop (PL), Fitbit Charge HR (FC), and SenseWear Armband Mini (SWA) (criterion measures: SWA for EE and a sleep log for SPT). The mean absolute percent error (MAPE) for EE was 13.0, 15.2, 15.5, 16.1, 16.2, 22.8, and 24.5% for PL, MF, FF, NFB, FC, JU, and VF, respectively. Mean absolute percent errors were calculated for SPT to be 4.0, 8.8, 10.2, 11.5, 12.9, 13.6, 17.5, and 21.61% for VF, FF, JU, FC, MF, SWA laying down, PL, and SWA, respectively. Concurrent validity was examined using equivalence testing on EE (equivalence zone: 2,889.7-3,531.9 kcal); 2 trackers fell short of falling in the zone: PL (2,714.4-3,164.8 kcal) and FC (2,473.8-3,066.5 kcal). For SPT (equivalence zone: 420.6-514.0 minutes), several monitors fell in the zone: PL (448.3-485.6 minutes), MS (442.8-492.2 minutes), and FF (427.7-486.7 minutes). This study suggests that the PL and FC provide a reasonable estimate of EE under free-living conditions. The PL, FC, and MF were the most valid monitors used for measuring SPT.

Scaffold-free Tissue Formation Under Real and Simulated Microgravity Conditions.

PubMed

Aleshcheva, Ganna; Bauer, Johann; Hemmersbach, Ruth; Slumstrup, Lasse; Wehland, Markus; Infanger, Manfred; Grimm, Daniela

2016-10-01

Scaffold-free tissue formation in microgravity is a new method in regenerative medicine and an important topic in Space Medicine. In this MiniReview, we focus on recent findings in the field of tissue engineering that were observed by exposing cells to real microgravity in space or to devices simulating to at least some extent microgravity conditions on Earth (ground-based facilities). Under both conditions - real and simulated microgravity - a part of the cultured cells of various populations detaches from the bottom of a culture flask. The cells form three-dimensional (3D) aggregates resembling the organs from which the cells have been derived. As spaceflights are rare and extremely expensive, cell culture under simulated microgravity allows more comprehensive and frequent studies on the scaffold-free 3D tissue formation in some aspects, as a number of publications have proven during the last two decades. In this MiniReview, we summarize data from our own studies and work from various researchers about tissue engineering of multi-cellular spheroids formed by cancer cells, tube formation by endothelial cells and cartilage formation by exposing the cells to ground-based facilities such as the 3D Random Positioning Machine (RPM), the 2D Fast-Rotating Clinostat (FRC) or the Rotating Wall Vessel (RWV). Subsequently, we investigated self-organization of 3D aggregates without scaffolds pursuing to enhance the frequency of 3D formation and to enlarge the size of the organ-like aggregates. The density of the monolayer exposed to real or simulated microgravity as well as the composition of the culture media revealed an impact on the results. Genomic and proteomic alterations were induced by simulated microgravity. Under microgravity conditions, adherent cells expressed other genes than cells grown in spheroids. In this MiniReview, the recent improvements in scaffold-free tissue formation are summarized and relationships between phenotypic and molecular appearance are

The global mean energy balance under cloud-free conditions

NASA Astrophysics Data System (ADS)

Wild, Martin; Hakuba, Maria; Folini, Dois; Ott, Patricia; Long, Charles

2017-04-01

A long standing problem of climate models is their overestimation of surface solar radiation not only under all-sky, but also under clear-sky conditions (Wild et al. 1995, Wild et al. 2006). This overestimation reduced over time in consecutive model generations due to the simulation of stronger atmospheric absorption. Here we analyze the clear sky fluxes of the latest climate model generation from the Coupled Model Intercomparison Project Phase 5 (CMIP5) against an expanded and updated set of direct observations from the Baseline Surface Radiation Network (BSRN). Clear sky climatologies from these sites have been composed based on the Long and Ackermann (2000) clear sky detection algorithm (Hakuba et al. 2017), and sampling issues when comparing with model simulated clear sky fluxes have been analyzed in Ott (2017). Overall, the overestimation of clear sky insolation in the CMIP5 models is now merely 1-2 Wm-2 in the multimodel mean, compared to 4 Wm-2 in CMIP3 and 6 Wm-2 in AMIPII (Wild et al. 2006). Still a considerable spread in the individual model biases is apparent, ranging from -2 Wm-2 to 10 Wm-2 when averaged over 53 globally distributed BSRN sites. This bias structure is used to infer best estimates for present day global mean clear sky insolation, following an approach developped in Wild et al. (2013, 2015, Clim. Dyn.) for all sky fluxes. Thereby the flux biases in the various models are linearly related to their respective global means. A best estimate can then be inferred from the linear regression at the intersect where the bias against the surface observations becomes zero. This way we obtain a best estimate of 247 Wm-2 for the global mean insolation at the Earth surface under cloud free conditions, and a global mean absorbed solar radiation of 214 Wm-2 in the cloud-free atmosphere, assuming a global mean surface albedo of 13.5%. Combined with a best estimate for the net influx of solar radiation at the Top of Atmosphere under cloud free conditions

The effect of encoding condition on free recall in Parkinson's disease: incidental and intentional memory are equally affected.

PubMed

Ellfolk, Ulla; Huurinainen, Salla; Joutsa, Juho; Karrasch, Mira

2012-01-01

Free recall memory deficits are common at early stages of Parkinson's disease (PD). As most studies have used intentional memory tasks, there is little information on how non-intentional, incidental encoding conditions affect memory performance in PD. We studied possible differences between PD patients and controls on free recall using incidental and intentional visual memory tasks. Free recall was examined in relation to attentive/executive functioning and subjective memory complaints. A total of 29 non-demented, medicated PD patients (age 60, disease duration 19 months) and 29 healthy controls (age 61) participated in the study. Incidental free recall was studied using a memory-modification of the Boston naming test (Memo-BNT) and intentional free recall with the 20 Objects test. There was a significant main effect for group due to worse free recall performances in the PD group. No statistically significant interaction between group and encoding condition was observed. The free recall deficit in the PD group was related to cognitive/psychomotor slowing, but not to attentive/executive task demands, or to subjective memory complaints. The results indicate that PD patients are impaired on free recall irrespective of encoding condition.

Inhibition of oxotremorine-induced desensitization of guinea-pig ileal longitudinal muscle in Ca2+-free conditions.

PubMed

Horio, S; Fukui, H

2001-02-01

The aim of this study was to investigate the differences between oxotremorine-induced and acetylcholine (ACh)-induced desensitization, particularly under Ca2+-free conditions, in guinea-pig ileal longitudinal muscle, and to elucidate the different mechanisms of desensitization that might exist between these two muscarinic agonists. Pretreatment of the tissue with 10(-7)-10(-5) M oxotremorine (desensitizing treatment) in normal Tyrode solution caused desensitization of the responses to ACh, as did the desensitizing treatment with ACh. However, Ca2+-free conditions significantly reduced oxotremorine-induced desensitization, contrary to the previous findings that Ca2+-free conditions enhanced ACh-induced desensitization. The desensitizing treatment with oxotremorine caused suppression of the responses to high K+ (tonic phase), as did the ACh treatment. Ca2+-free conditions removed this suppression, whereasthis condition enhanced ACh-induced suppression of the K+ response. A protein kinase C inhibitor, 1-(5-isoquinolinesulfonyl)-2-methylpiperazine (10(-4) M) had no effect on oxotremorine-induced desensitization of the ACh response. The results suggest that a voltage-gated Ca2+ channel was involved in oxotremorine-induced desensitization, as in ACh-induced desensitization, but that the process of inactivation of Ca2+ channels was different between oxotremorine and ACh, and that oxotremorine-induced desensitization was due not only to Ca2+ channel, but also to other unknown factors. Protein kinase C did not participate in oxotremorine-induced desensitization.

Ising antiferromagnet on a finite triangular lattice with free boundary conditions

NASA Astrophysics Data System (ADS)

Kim, Seung-Yeon

2015-11-01

The exact integer values for the density of states of the Ising model on an equilateral triangular lattice with free boundary conditions are evaluated up to L = 24 spins on a side for the first time by using the microcanonical transfer matrix. The total number of states is 2 N s = 2300 ≈ 2.037 × 1090 for L = 24, where N s = L( L+1)/2 is the number of spins. Classifying all 2300 spin states according to their energy values is an enormous work. From the density of states, the exact partition function zeros in the complex temperature plane of the triangular-lattice Ising model are evaluated. Using the density of states and the partition function zeros, we investigate the properties of the triangularlattice Ising antiferromagnet. The scaling behavior of the ground-state entropy and the form of the correlation length at T = 0 are studied for the triangular-lattice Ising antiferromagnet with free boundary conditions. Also, the scaling behavior of the Fisher edge singularity is investigated.

Microcarrier-based platforms for in vitro expansion and differentiation of human pluripotent stem cells in bioreactor culture systems.

PubMed

Badenes, Sara M; Fernandes, Tiago G; Rodrigues, Carlos A V; Diogo, Maria Margarida; Cabral, Joaquim M S

2016-09-20

Human pluripotent stem cells (hPSC) have attracted a great attention as an unlimited source of cells for cell therapies and other in vitro biomedical applications such as drug screening, toxicology assays and disease modeling. The implementation of scalable culture platforms for the large-scale production of hPSC and their derivatives is mandatory to fulfill the requirement of obtaining large numbers of cells for these applications. Microcarrier technology has been emerging as an effective approach for the large scale ex vivo hPSC expansion and differentiation. This review presents recent achievements in hPSC microcarrier-based culture systems and discusses the crucial aspects that influence the performance of these culture platforms. Recent progress includes addressing chemically-defined culture conditions for manufacturing of hPSC and their derivatives, with the development of xeno-free media and microcarrier coatings to meet good manufacturing practice (GMP) quality requirements. Finally, examples of integrated platforms including hPSC expansion and directed differentiation to specific lineages are also presented in this review. Copyright © 2016 Elsevier B.V. All rights reserved.

Tissue Source and Cell Expansion Condition Influence Phenotypic Changes of Adipose-Derived Stem Cells

PubMed Central

Mangum, Lauren H.; Stone, Randolph; Wrice, Nicole L.; Larson, David A.; Florell, Kyle F.; Christy, Barbara A.; Herzig, Maryanne C.; Cap, Andrew P.

2017-01-01

Stem cells derived from the subcutaneous adipose tissue of debrided burned skin represent an appealing source of adipose-derived stem cells (ASCs) for regenerative medicine. Traditional tissue culture uses fetal bovine serum (FBS), which complicates utilization of ASCs in human medicine. Human platelet lysate (hPL) is one potential xeno-free, alternative supplement for use in ASC culture. In this study, adipogenic and osteogenic differentiation in media supplemented with 10% FBS or 10% hPL was compared in human ASCs derived from abdominoplasty (HAP) or from adipose associated with debrided burned skin (BH). Most (95–99%) cells cultured in FBS were stained positive for CD73, CD90, CD105, and CD142. FBS supplementation was associated with increased triglyceride content and expression of adipogenic genes. Culture in hPL significantly decreased surface staining of CD105 by 31% and 48% and CD142 by 27% and 35% in HAP and BH, respectively (p < 0.05). Culture of BH-ASCs in hPL also increased expression of markers of osteogenesis and increased ALP activity. These data indicate that application of ASCs for wound healing may be influenced by ASC source as well as culture conditions used to expand them. As such, these factors must be taken into consideration before ASCs are used for regenerative purposes. PMID:29138638

Engineering large cartilage tissues using dynamic bioreactor culture at defined oxygen conditions.

PubMed

Daly, Andrew C; Sathy, Binulal N; Kelly, Daniel J

2018-01-01

Mesenchymal stem cells maintained in appropriate culture conditions are capable of producing robust cartilage tissue. However, gradients in nutrient availability that arise during three-dimensional culture can result in the development of spatially inhomogeneous cartilage tissues with core regions devoid of matrix. Previous attempts at developing dynamic culture systems to overcome these limitations have reported suppression of mesenchymal stem cell chondrogenesis compared to static conditions. We hypothesize that by modulating oxygen availability during bioreactor culture, it is possible to engineer cartilage tissues of scale. The objective of this study was to determine whether dynamic bioreactor culture, at defined oxygen conditions, could facilitate the development of large, spatially homogeneous cartilage tissues using mesenchymal stem cell laden hydrogels. A dynamic culture regime was directly compared to static conditions for its capacity to support chondrogenesis of mesenchymal stem cells in both small and large alginate hydrogels. The influence of external oxygen tension on the response to the dynamic culture conditions was explored by performing the experiment at 20% O 2 and 3% O 2 . At 20% O 2 , dynamic culture significantly suppressed chondrogenesis in engineered tissues of all sizes. In contrast, at 3% O 2 dynamic culture significantly enhanced the distribution and amount of cartilage matrix components (sulphated glycosaminoglycan and collagen II) in larger constructs compared to static conditions. Taken together, these results demonstrate that dynamic culture regimes that provide adequate nutrient availability and a low oxygen environment can be employed to engineer large homogeneous cartilage tissues. Such culture systems could facilitate the scaling up of cartilage tissue engineering strategies towards clinically relevant dimensions.

Communication: modeling charge-sign asymmetric solvation free energies with nonlinear boundary conditions.

PubMed

Bardhan, Jaydeep P; Knepley, Matthew G

2014-10-07

We show that charge-sign-dependent asymmetric hydration can be modeled accurately using linear Poisson theory after replacing the standard electric-displacement boundary condition with a simple nonlinear boundary condition. Using a single multiplicative scaling factor to determine atomic radii from molecular dynamics Lennard-Jones parameters, the new model accurately reproduces MD free-energy calculations of hydration asymmetries for: (i) monatomic ions, (ii) titratable amino acids in both their protonated and unprotonated states, and (iii) the Mobley "bracelet" and "rod" test problems [D. L. Mobley, A. E. Barber II, C. J. Fennell, and K. A. Dill, "Charge asymmetries in hydration of polar solutes," J. Phys. Chem. B 112, 2405-2414 (2008)]. Remarkably, the model also justifies the use of linear response expressions for charging free energies. Our boundary-element method implementation demonstrates the ease with which other continuum-electrostatic solvers can be extended to include asymmetry.

Superior Red Blood Cell Generation from Human Pluripotent Stem Cells Through a Novel Microcarrier-Based Embryoid Body Platform.

PubMed

Sivalingam, Jaichandran; Lam, Alan Tin-Lun; Chen, Hong Yu; Yang, Bin Xia; Chen, Allen Kuan-Liang; Reuveny, Shaul; Loh, Yuin-Han; Oh, Steve Kah-Weng

2016-08-01

In vitro generation of red blood cells (RBCs) from human embryonic stem cells and human induced pluripotent stem cells appears to be a promising alternate approach to circumvent shortages in donor-derived blood supplies for clinical applications. Conventional methods for hematopoietic differentiation of human pluripotent stem cells (hPSC) rely on embryoid body (EB) formation and/or coculture with xenogeneic cell lines. However, most current methods for hPSC expansion and EB formation are not amenable for scale-up to levels required for large-scale RBC generation. Moreover, differentiation methods that rely on xenogenic cell lines would face obstacles for future clinical translation. In this study, we report the development of a serum-free and chemically defined microcarrier-based suspension culture platform for scalable hPSC expansion and EB formation. Improved survival and better quality EBs generated with the microcarrier-based method resulted in significantly improved mesoderm induction and, when combined with hematopoietic differentiation, resulted in at least a 6-fold improvement in hematopoietic precursor expansion, potentially culminating in a 80-fold improvement in the yield of RBC generation compared to a conventional EB-based differentiation method. In addition, we report efficient terminal maturation and generation of mature enucleated RBCs using a coculture system that comprised primary human mesenchymal stromal cells. The microcarrier-based platform could prove to be an appealing strategy for future scale-up of hPSC culture, EB generation, and large-scale generation of RBCs under defined and xeno-free conditions.

Regulated production of free radicals by the mitochondrial electron transport chain: Cardiac ischemic preconditioning.

PubMed

Matsuzaki, Satoshi; Szweda, Pamela A; Szweda, Luke I; Humphries, Kenneth M

2009-11-30

Excessive production of free radicals by mitochondria is associated with, and likely contributes to, the progression of numerous pathological conditions. Nevertheless, the production of free radicals by the mitochondria may have important biological functions under normal or stressed conditions by activating or modulating redox-sensitive cellular signaling pathways. This raises the intriguing possibility that regulated mitochondrial free radical production occurs via mechanisms that are distinct from pathologies associated with oxidative damage. Indeed, the capacity of mitochondria to produce free radicals in a limited manner may play a role in ischemic preconditioning, the phenomenon whereby short bouts of ischemia protect from subsequent prolonged ischemia and reperfusion. Ischemic preconditioning can thus serve as an important model system for defining regulatory mechanisms that allow for transient, signal-inducing, production of free radicals by mitochondria. Defining how these mechanism(s) occur will provide insight into therapeutic approaches that minimize oxidative damage without altering normal cellular redox biology. The aim of this review is to present and discuss evidence for the regulated production of superoxide by the electron transport chain within the ischemic preconditioning paradigm of redox regulation.

Measurements in Transitional Boundary Layers Under High Free-Stream Turbulence and Strong Acceleration Conditions

NASA Technical Reports Server (NTRS)

Volino, Ralph J.; Simon, Terrence W.

1995-01-01

Measurements from transitional, heated boundary layers along a concave-curved test wall are presented and discussed. A boundary layer subject to low free-stream turbulence intensity (FSTI), which contains stationary streamwise (Gortler) vortices, is documented. The low FSTI measurements are followed by measurements in boundary layers subject to high (initially 8%) free-stream turbulence intensity and moderate to strong streamwise acceleration. Conditions were chosen to simulate those present on the downstream half of the pressure side of a gas turbine airfoil. Mean flow characteristics as well as turbulence statistics, including the turbulent shear stress, turbulent heat flux, and turbulent Prandtl number, are documented. A technique called "octant analysis" is introduced and applied to several cases from the literature as well as to data from the present study. Spectral analysis was applied to describe the effects of turbulence scales of different sizes during transition. To the authors'knowledge, this is the first detailed documentation of boundary layer transition under such high free-stream turbulence conditions.

Communication: Modeling charge-sign asymmetric solvation free energies with nonlinear boundary conditions

DOE Office of Scientific and Technical Information (OSTI.GOV)

Bardhan, Jaydeep P.; Knepley, Matthew G.

2014-10-07

We show that charge-sign-dependent asymmetric hydration can be modeled accurately using linear Poisson theory after replacing the standard electric-displacement boundary condition with a simple nonlinear boundary condition. Using a single multiplicative scaling factor to determine atomic radii from molecular dynamics Lennard-Jones parameters, the new model accurately reproduces MD free-energy calculations of hydration asymmetries for: (i) monatomic ions, (ii) titratable amino acids in both their protonated and unprotonated states, and (iii) the Mobley “bracelet” and “rod” test problems [D. L. Mobley, A. E. Barber II, C. J. Fennell, and K. A. Dill, “Charge asymmetries in hydration of polar solutes,” J. Phys.more » Chem. B 112, 2405–2414 (2008)]. Remarkably, the model also justifies the use of linear response expressions for charging free energies. Our boundary-element method implementation demonstrates the ease with which other continuum-electrostatic solvers can be extended to include asymmetry.« less

Communication: Modeling charge-sign asymmetric solvation free energies with nonlinear boundary conditions

PubMed Central

Bardhan, Jaydeep P.; Knepley, Matthew G.

2014-01-01

We show that charge-sign-dependent asymmetric hydration can be modeled accurately using linear Poisson theory after replacing the standard electric-displacement boundary condition with a simple nonlinear boundary condition. Using a single multiplicative scaling factor to determine atomic radii from molecular dynamics Lennard-Jones parameters, the new model accurately reproduces MD free-energy calculations of hydration asymmetries for: (i) monatomic ions, (ii) titratable amino acids in both their protonated and unprotonated states, and (iii) the Mobley “bracelet” and “rod” test problems [D. L. Mobley, A. E. Barber II, C. J. Fennell, and K. A. Dill, “Charge asymmetries in hydration of polar solutes,” J. Phys. Chem. B 112, 2405–2414 (2008)]. Remarkably, the model also justifies the use of linear response expressions for charging free energies. Our boundary-element method implementation demonstrates the ease with which other continuum-electrostatic solvers can be extended to include asymmetry. PMID:25296776

"Dermatitis" defined.

PubMed

Smith, Suzanne M; Nedorost, Susan T

2010-01-01

The term "dermatitis" can be defined narrowly or broadly, clinically or histologically. A common and costly condition, dermatitis is underresourced compared to other chronic skin conditions. The lack of a collectively understood definition of dermatitis and its subcategories could be the primary barrier. To investigate how dermatologists define the term "dermatitis" and determine if a consensus on the definition of this term and other related terms exists. A seven-question survey of dermatologists nationwide was conducted. Of respondents (n  =  122), half consider dermatitis to be any inflammation of the skin. Nearly half (47.5%) use the term interchangeably with "eczema." Virtually all (> 96%) endorse the subcategory "atopic" under the terms "dermatitis" and "eczema," but the subcategories "contact," "drug hypersensitivity," and "occupational" are more highly endorsed under the term "dermatitis" than under the term "eczema." Over half (55.7%) personally consider "dermatitis" to have a broad meaning, and even more (62.3%) believe that dermatologists as a whole define the term broadly. There is a lack of consensus among experts in defining dermatitis, eczema, and their related subcategories.

Model-Free Conditional Independence Feature Screening For Ultrahigh Dimensional Data.

PubMed

Wang, Luheng; Liu, Jingyuan; Li, Yong; Li, Runze

2017-03-01

Feature screening plays an important role in ultrahigh dimensional data analysis. This paper is concerned with conditional feature screening when one is interested in detecting the association between the response and ultrahigh dimensional predictors (e.g., genetic makers) given a low-dimensional exposure variable (such as clinical variables or environmental variables). To this end, we first propose a new index to measure conditional independence, and further develop a conditional screening procedure based on the newly proposed index. We systematically study the theoretical property of the proposed procedure and establish the sure screening and ranking consistency properties under some very mild conditions. The newly proposed screening procedure enjoys some appealing properties. (a) It is model-free in that its implementation does not require a specification on the model structure; (b) it is robust to heavy-tailed distributions or outliers in both directions of response and predictors; and (c) it can deal with both feature screening and the conditional screening in a unified way. We study the finite sample performance of the proposed procedure by Monte Carlo simulations and further illustrate the proposed method through two real data examples.

Plate waste of adults in the United States measured in free-living conditions

PubMed Central

Allen, H. Raymond

2018-01-01

We analyze food-item level data collected from 50 adults from the United States using the Remote Food Photography Method® to provide the first estimates of plate waste gathered from adults across multiple consecutive meals and days in free-living conditions, and during laboratory-based meals with fixed food items and quantities. We find average plate waste in free-living conditions is 5.6 grams (7.7 kcals) per item and that 3.3% of all food selected is returned as plate waste, where the percent waste figure is substantially lower than previously published plate waste estimates gathered primarily from dine-out settings in the United States such as buffets and institutional settings with limited-choice meals (e.g., school cafeterias). Plate waste from the same participants during the laboratory-based meals is significantly higher with an average of 203.2 grams of solid plate waste per meal (531.3 kcals) or 39.1% of the food provided, which is similar to the plate waste percentages found reported in some school cafeteria settings. The amount of plate waste generated in free-living conditions is significantly positively associated with portion size selected for an item. In a multivariate analysis that controls for macronutrient profile, items selected from the vegetables, fats/oils/dressings, and grains categories are associated with significantly greater amounts of plate waste per item. We find no significant associations between free-living plate waste and gender, age, race or body mass index but find that women leave more plate waste in the lab meal where portion sizes are pre-determined by the researcher and similar for all respondents. We discuss possible implications of these findings for programs focused on reducing plate waste and food waste among consumers. PMID:29444094

Modelling the complete operation of a free-piston shock tunnel for a low enthalpy condition

NASA Astrophysics Data System (ADS)

McGilvray, M.; Dann, A. G.; Jacobs, P. A.

2013-07-01

Only a limited number of free-stream flow properties can be measured in hypersonic impulse facilities at the nozzle exit. This poses challenges for experimenters when subsequently analysing experimental data obtained from these facilities. Typically in a reflected shock tunnel, a simple analysis that requires small amounts of computational resources is used to calculate quasi-steady gas properties. This simple analysis requires initial fill conditions and experimental measurements in analytical calculations of each major flow process, using forward coupling with minor corrections to include processes that are not directly modeled. However, this simplistic approach leads to an unknown level of discrepancy to the true flow properties. To explore the simple modelling techniques accuracy, this paper details the use of transient one and two-dimensional numerical simulations of a complete facility to obtain more refined free-stream flow properties from a free-piston reflected shock tunnel operating at low-enthalpy conditions. These calculations were verified by comparison to experimental data obtained from the facility. For the condition and facility investigated, the test conditions at nozzle exit produced with the simple modelling technique agree with the time and space averaged results from the complete facility calculations to within the accuracy of the experimental measurements.

Effects of two solvent conditions on the free energy landscape of the BBL peripheral subunit binding domain.

PubMed

Liu, Hanzhong; Huo, Shuanghong

2012-01-12

BBL is a small independently folding domain with two main parallel helices. The experiment of C(α) secondary shifts has shown that changing the pH from ~7 to ~5 results in the reduced helicity at the C-terminus of helix 2. Combining constant pH molecular dynamics with replica exchange, we sampled the protein conformation space and protonation states extensively under a neutral pH condition and an acidic condition. Our results reveal that the solvent conditions influence the free energy landscape. Under the neutral pH condition, the denatured state and the native state are well separated. The condition of the acidic pH reshapes the free energy surface, leading to a broadly populated denatured-state basin and a low free energy barrier between the denatured state and the native state. The acidic pH shifts the equilibrium between the denatured state and the native state in favor of the denatured state. Caution must be used to interpret experimental data under the acidic condition because the contribution of the denatured state is significant. Our simulation results are supported by the fact that the calculated chemical shifts are in good agreement with the experiment data.

A comparison of shear bond strength of orthodontic brackets bonded with four different orthodontic adhesives

PubMed Central

Sharma, Sudhir; Tandon, Pradeep; Nagar, Amit; Singh, Gyan P; Singh, Alka; Chugh, Vinay K

2014-01-01

Objectives: The objective of this study is to compare the shear bond strength (SBS) of stainless steel (SS) orthodontic brackets bonded with four different orthodontic adhesives. Materials and Methods: Eighty newly extracted premolars were bonded to 0.022 SS brackets (Ormco, Scafati, Italy) and equally divided into four groups based on adhesive used: (1) Rely-a-Bond (self-cure adhesive, Reliance Orthodontic Product, Inc., Illinois, USA), (2) Transbond XT (light-cure adhesive, 3M Unitek, CA, USA), (3) Transbond Plus (sixth generation self-etch primer, 3M Unitek, CA, USA) with Transbond XT (4) Xeno V (seventh generation self-etch primer, Dentsply, Konstanz, Germany) with Xeno Ortho (light-cure adhesive, Dentsply, Konstanz, Germany) adhesive. Brackets were debonded with a universal testing machine (Model No. 3382 Instron Corp., Canton, Mass, USA). The adhesive remnant index (ARI) was recordedIn addition, the conditioned enamel surfaces were observed under a scanning electron microscope (SEM). Results: Transbond XT (15.49 MPa) attained the highest bond strength. Self-etching adhesives (Xeno V, 13.51 MPa; Transbond Plus, 11.57 MPa) showed clinically acceptable SBS values and almost clean enamel surface after debonding. The analysis of variance (F = 11.85, P < 0.0001) and Chi-square (χ2 = 18.16, P < 0.05) tests revealed significant differences among groups. The ARI score of 3 (i.e., All adhesives left on the tooth) to be the most prevalent in Transbond XT (40%), followed by Rely-a-Bond (30%), Transbond Plus with Transbond XT (15%), and Xeno V with Xeno Ortho (10%). Under SEM, enamel surfaces after debonding of the brackets appeared porous when an acid-etching process was performed on the surfaces of Rely-a-Bond and Transbond XT, whereas with self-etching primers enamel presented smooth and almost clean surfaces (Transbond Plus and Xeno V group). Conclusion: All adhesives yielded SBS values higher than the recommended bond strength (5.9-7–8 MPa), Seventh generation

Subtle effect of Xenos vesparum (Xenidae, Strepsiptera) on the reproductive apparatus of its male host: Parasite or parasitoid?

PubMed

Beani, L; Marchini, D; Cappa, F; Petrocelli, I; Gottardo, M; Manfredini, F; Giusti, F; Dallai, R

2017-08-01

Parasitic castration is an adaptive strategy where parasites usurp the hosts' reproductive physiology to complete their life cycle. The alterations in the host traits vary in their magnitude, from subtle changes in the host morpho-physiology and behaviour to the production of complex aberrant phenotypes, which often depend on the host gender. The strepsipteran macroparasite Xenos vesparum induces dramatic behavioural and physiological changes in its female host, the paper wasp Polistes dominula, while its effect on the male phenotype is largely unknown. In this study we investigated how a single X. vesparum parasite influences the functional morphology of P. dominula male reproductive apparatus. We performed morphometry and ultrastructure characterization of corpora allata, testes, seminal vesicles and accessory glands in parasitized and unparasitized males, and also in young and old males to control for the effect of age on the natural deterioration of these organs. Our results show that age significantly affects the development of male reproductive apparatus. A low parasite load - one parasite per host is the common prevalence in the field - has only a marginal impact on the reproductive morphology of P. dominula males, affecting quantitatively but not qualitatively the protein content of male accessory glands. Thus, in male P. dominula wasps, X. vesparum appears to behave as a true "parasite", in clear opposition to the role of "parasitoid" that it takes in female hosts where castration causes the reproductive death. Copyright © 2017 Elsevier Ltd. All rights reserved.

Beam Conditioning and Harmonic Generation in Free ElectronLasers

DOE Office of Scientific and Technical Information (OSTI.GOV)

Charman, A.E.; Penn, G.; Wolski, A.

2004-07-05

The next generation of large-scale free-electron lasers (FELs) such as Euro-XFEL and LCLS are to be devices which produce coherent X-rays using Self-Amplified Spontaneous Emission (SASE). The performance of these devices is limited by the spread in longitudinal velocities of the beam. In the case where this spread arises primarily from large transverse oscillation amplitudes, beam conditioning can significantly enhance FEL performance. Future X-ray sources may also exploit harmonic generation starting from laser-seeded modulation. Preliminary analysis of such devices is discussed, based on a novel trial-function/variational-principle approach, which shows good agreement with more lengthy numerical simulations.

Heat generation in aircraft tires under free rolling conditions

NASA Technical Reports Server (NTRS)

Clark, S. K.; Dodge, R. N.

1982-01-01

A method was developed for calculating the internal temperature distribution in an aircraft tire while free rolling under load. The method uses an approximate stress analysis of each point in the tire as it rolls through the contact patch, and from this stress change the mechanical work done on each volume element may be obtained and converted into a heat release rate through a knowledge of material characteristics. The tire cross-section is then considered as a body with internal heat generation, and the diffusion equation is solved numerically with appropriate boundary conditions of the wheel and runway surface. Comparison with data obtained with buried thermocouples in tires shows good agreement.

Effects of Anorectic Drugs on Food Intake under Progressive-Ratio and Free-Access Conditions in Rats

ERIC Educational Resources Information Center

LeSage, Mark G.; Stafford, David; Glowa, John R.

2004-01-01

The effects of two anorectic drugs, dexfenfluramine and phentermine, on food intake under different food-access conditions were examined. Experiment 1 compared the effects of these drugs on food intake under a progressive-ratio (PR) schedule and free-access conditions. Dexfenfluramine decreased food intake under both conditions, but the doses…

Optimizing Extraction Conditions of Free and Bound Phenolic Compounds from Rice By-Products and Their Antioxidant Effects.

PubMed

Irakli, Maria; Kleisiaris, Fotis; Kadoglidou, Kalliopi; Katsantonis, Dimitrios

2018-06-13

Rice by-products are extensively abundant agricultural wastes from the rice industry. This study was designed to optimize experimental conditions for maximum recovery of free and bound phenolic compounds from rice by-products. Optimized conditions were determined using response surface methodology based on total phenolic content (TPC), ABTS radical scavenging activity and ferric reducing power (FRAP). A Box-Behnken design was used to investigate the effects of ethanol concentration, extraction time and temperature, and NaOH concentration, hydrolysis time and temperature for free and bound fractions, respectively. The optimal conditions for the free phenolics were 41⁻56%, 40 °C, 10 min, whereas for bound phenolics were 2.5⁻3.6 M, 80 °C, 120 min. Under these conditions free TPC, ABTS and FRAP values in the bran were approximately 2-times higher than in the husk. However, bound TPC and FRAP values in the husk were 1.9- and 1.2-times higher than those in the bran, respectively, while bran fraction observed the highest ABTS value. Ferulic acid was most evident in the bran, whereas p -coumaric acid was mostly found in the husk. Findings from this study demonstrates that rice by-products could be exploited as valuable sources of bioactive components that could be used as ingredients of functional food and nutraceuticals.

Epithelial Coculture and l-Lactate Promote Growth of Helicobacter cinaedi under H2-Free Aerobic Conditions

PubMed Central

Taniguchi, Takako; Misawa, Naoaki

2016-01-01

ABSTRACT Helicobacter cinaedi is an emerging opportunistic pathogen associated with infections of diverse anatomic sites. Nevertheless, the species demonstrates fastidious axenic growth; it has been described as requiring a microaerobic atmosphere, along with a strong preference for supplemental H2 gas. In this context, we examined the hypothesis that in vitro growth of H. cinaedi could be enhanced by coculture with human epithelial cells. When inoculated (in Ham's F12 medium) over Caco-2 monolayers, the type strain (ATCC BAA-847) gained the ability to proliferate under H2-free aerobic conditions. Identical results were observed during coculture with several other monolayer types (LS-174T, AGS, and HeLa). Under chemically defined conditions, 40 amino acids and carboxylates were screened for their effect on the organism's atmospheric requirements. Several molecules promoted H2-free aerobic proliferation, although it occurred most prominently with millimolar concentrations of l-lactate. The growth response of H. cinaedi to Caco-2 cells and l-lactate was confirmed with a collection of 12 human-derived clinical strains. mRNA sequencing was next performed on the type strain under various growth conditions. In addition to providing a whole-transcriptome profile of H. cinaedi, this analysis demonstrated strong constitutive expression of the l-lactate utilization locus, as well as differential transcription of terminal respiratory proteins as a function of Caco-2 coculture and l-lactate supplementation. Overall, these findings challenge traditional views of H. cinaedi as an obligate microaerophile. IMPORTANCE H. cinaedi is an increasingly recognized pathogen in people with compromised immune systems. Atypical among other members of its bacterial class, H. cinaedi has been associated with infections of diverse anatomic sites. Growing H. cineadi in the laboratory is quite difficult, due in large part to the need for a specialized atmosphere. The suboptimal growth of H

Preserving enzymatic activity and enhancing biochemical stability of glutathione transferase by soluble additives under free and tethered conditions.

PubMed

Karamitros, Christos S; Labrou, Nikolaos E

2017-09-01

In the present study, we report the effect of four different soluble additives (sucrose, lactitol, superfloc c577, and dextran sulfate) on the stability of glutathione transferase 1 enzyme from Zea mays (ZmGSTF1-1) under free and tethered conditions at 4 and 25 °C. Among all additives, the best stabilizing effects were observed in the case of superfloc c577 and sucrose at both tested temperatures, yet at distinct concentrations at each condition. Those two stabilizing agents were further combined and potential positive synergistic effects were investigated. In addition, we assessed the long-term storage and operational stability of ZmGSTF1-1 under tethered conditions in the presence of additives, which provided the most conducive effects on its stability under free conditions. Our results strongly suggest that the presence of additives may be beneficial to the stability of the enzyme under both free and tethered conditions. Thermodynamic analysis of the free enzyme in the presence of sucrose, which exhibited the best stabilizing effect at both temperatures, shed light on the possible mechanism of action. Given the considerable importance of the development of GST-based biosensors with prolonged stability, the present work may be of general interest to researchers in the field of applied enzymology. © 2016 International Union of Biochemistry and Molecular Biology, Inc.

Stabilizing effect of resistivity towards ELM-free H-mode discharge in lithium-conditioned NSTX

NASA Astrophysics Data System (ADS)

Banerjee, Debabrata; Zhu, Ping; Maingi, Rajesh

2017-07-01

Linear stability analysis of the national spherical torus experiment (NSTX) Li-conditioned ELM-free H-mode equilibria is carried out in the context of the extended magneto-hydrodynamic (MHD) model in NIMROD. The purpose is to investigate the physical cause behind edge localized mode (ELM) suppression in experiment after the Li-coating of the divertor and the first wall of the NSTX tokamak. Besides ideal MHD modeling, including finite-Larmor radius effect and two-fluid Hall and electron diamagnetic drift contributions, a non-ideal resistivity model is employed, taking into account the increase of Z eff after Li-conditioning in ELM-free H-mode. Unlike an earlier conclusion from an eigenvalue code analysis of these equilibria, NIMROD results find that after reduced recycling from divertor plates, profile modification is necessary but insufficient to explain the mechanism behind complete ELMs suppression in ideal two-fluid MHD. After considering the higher plasma resistivity due to higher Z eff, the complete stabilization could be explained. A thorough analysis of both pre-lithium ELMy and with-lithium ELM-free cases using ideal and non-ideal MHD models is presented, after accurately including a vacuum-like cold halo region in NIMROD to investigate ELMs.

Defined Engineered Human Myocardium with Advanced Maturation for Applications in Heart Failure Modelling and Repair

PubMed Central

Tiburcy, Malte; Hudson, James E.; Balfanz, Paul; Schlick, Susanne; Meyer, Tim; Liao, Mei-Ling Chang; Levent, Elif; Raad, Farah; Zeidler, Sebastian; Wingender, Edgar; Riegler, Johannes; Wang, Mouer; Gold, Joseph D.; Kehat, Izhak; Wettwer, Erich; Ravens, Ursula; Dierickx, Pieterjan; van Laake, Linda W.; Goumans, Marie Jose; Khadjeh, Sara; Toischer, Karl; Hasenfuss, Gerd; Couture, Larry A.; Unger, Andreas; Linke, Wolfgang A.; Araki, Toshiyuki; Neel, Benjamin; Keller, Gordon; Gepstein, Lior; Wu, Joseph C.; Zimmermann, Wolfram-Hubertus

2017-01-01

Background Advancing structural and functional maturation of stem cell-derived cardiomyocytes remains a key challenge for applications in disease modelling, drug screening, and heart repair. Here, we sought to advance cardiomyocyte maturation in engineered human myocardium (EHM) towards an adult phenotype under defined conditions. Methods We systematically investigated cell composition, matrix and media conditions to generate EHM from embryonic and induced pluripotent stem cell-derived cardiomyocytes and fibroblasts with organotypic functionality under serum-free conditions. We employed morphological, functional, and transcriptome analyses to benchmark maturation of EHM. Results EHM demonstrated important structural and functional properties of postnatal myocardium, including: (1) rod-shaped cardiomyocytes with M-bands assembled as a functional syncytium; (2) systolic twitch forces at a similar level as observed in bona fide postnatal myocardium; (3) a positive force-frequency-response; (4) inotropic responses to β-adrenergic stimulation mediated via canonical β1- and β2-adrenoceptor signaling pathways; and (5) evidence for advanced molecular maturation by transcriptome profiling. EHM responded to chronic catecholamine toxicity with contractile dysfunction, cardiomyocyte hypertrophy, cardiomyocyte death, and NT-proBNP release; all are classical hallmarks of heart failure. Additionally, we demonstrate scalability of EHM according to anticipated clinical demands for cardiac repair. Conclusions We provide proof-of-concept for a universally applicable technology for the engineering of macro-scale human myocardium for disease modelling and heart repair from embryonic and induced pluripotent stem cell-derived cardiomyocytes under defined, serum-free conditions. PMID:28167635

Defined Engineered Human Myocardium With Advanced Maturation for Applications in Heart Failure Modeling and Repair.

PubMed

Tiburcy, Malte; Hudson, James E; Balfanz, Paul; Schlick, Susanne; Meyer, Tim; Chang Liao, Mei-Ling; Levent, Elif; Raad, Farah; Zeidler, Sebastian; Wingender, Edgar; Riegler, Johannes; Wang, Mouer; Gold, Joseph D; Kehat, Izhak; Wettwer, Erich; Ravens, Ursula; Dierickx, Pieterjan; van Laake, Linda W; Goumans, Marie Jose; Khadjeh, Sara; Toischer, Karl; Hasenfuss, Gerd; Couture, Larry A; Unger, Andreas; Linke, Wolfgang A; Araki, Toshiyuki; Neel, Benjamin; Keller, Gordon; Gepstein, Lior; Wu, Joseph C; Zimmermann, Wolfram-Hubertus

2017-05-09

Advancing structural and functional maturation of stem cell-derived cardiomyocytes remains a key challenge for applications in disease modeling, drug screening, and heart repair. Here, we sought to advance cardiomyocyte maturation in engineered human myocardium (EHM) toward an adult phenotype under defined conditions. We systematically investigated cell composition, matrix, and media conditions to generate EHM from embryonic and induced pluripotent stem cell-derived cardiomyocytes and fibroblasts with organotypic functionality under serum-free conditions. We used morphological, functional, and transcriptome analyses to benchmark maturation of EHM. EHM demonstrated important structural and functional properties of postnatal myocardium, including: (1) rod-shaped cardiomyocytes with M bands assembled as a functional syncytium; (2) systolic twitch forces at a similar level as observed in bona fide postnatal myocardium; (3) a positive force-frequency response; (4) inotropic responses to β-adrenergic stimulation mediated via canonical β 1 - and β 2 -adrenoceptor signaling pathways; and (5) evidence for advanced molecular maturation by transcriptome profiling. EHM responded to chronic catecholamine toxicity with contractile dysfunction, cardiomyocyte hypertrophy, cardiomyocyte death, and N-terminal pro B-type natriuretic peptide release; all are classical hallmarks of heart failure. In addition, we demonstrate the scalability of EHM according to anticipated clinical demands for cardiac repair. We provide proof-of-concept for a universally applicable technology for the engineering of macroscale human myocardium for disease modeling and heart repair from embryonic and induced pluripotent stem cell-derived cardiomyocytes under defined, serum-free conditions. © 2017 American Heart Association, Inc.

Modeling the effect of initial and free-stream conditions on circular wakes

NASA Astrophysics Data System (ADS)

Lewalle, Jacques

A cascade-transport model is applied to study the effect of initial and free-stream conditions on circular waves. The role of the very-large-eddies (VLEs) is shown and used to derive a new understanding of wakes and their lack of universality. Computational results are reported which show that the VLEs are a determining factor in the development of self-preserving solutions for the axisymmetric wake.

Spatiotemporal characteristics of severe dry and wet conditions in the Free State Province, South Africa

NASA Astrophysics Data System (ADS)

Mbiriri, M.; Mukwada, G.; Manatsa, D.

2018-02-01

This paper assesses the spatiotemporal characteristics of agricultural droughts and wet conditions in the Free State Province of South Africa for the period between 1960 and 2013. Since agriculturally, the Free State Province is considered the bread basket of the country, understanding the variability of drought and wet conditions becomes necessary. The Standardised Precipitation Index (SPI) computed from gridded monthly precipitation data was used to assess the rainfall extreme conditions. Hot spot analysis was used to divide the province into five homogenous clusters where the spatiotemporal characteristics for each cluster were analysed. The results show a west to east increase in seasonal average total precipitation. However, the eastern part of the province demonstrates higher occurrences of droughts, with SPI ≤ - 1.282. This is despite the observation that the region shows a recent increase in droughts unlike the western region. It is also noted that significant differences in drought/wet intensities between clusters are more pronounced during the early compared to the late summer period.

Free-form geometric modeling by integrating parametric and implicit PDEs.

PubMed

Du, Haixia; Qin, Hong

2007-01-01

Parametric PDE techniques, which use partial differential equations (PDEs) defined over a 2D or 3D parametric domain to model graphical objects and processes, can unify geometric attributes and functional constraints of the models. PDEs can also model implicit shapes defined by level sets of scalar intensity fields. In this paper, we present an approach that integrates parametric and implicit trivariate PDEs to define geometric solid models containing both geometric information and intensity distribution subject to flexible boundary conditions. The integrated formulation of second-order or fourth-order elliptic PDEs permits designers to manipulate PDE objects of complex geometry and/or arbitrary topology through direct sculpting and free-form modeling. We developed a PDE-based geometric modeling system for shape design and manipulation of PDE objects. The integration of implicit PDEs with parametric geometry offers more general and arbitrary shape blending and free-form modeling for objects with intensity attributes than pure geometric models.

LQG/LTR optimal attitude control of small flexible spacecraft using free-free boundary conditions

NASA Astrophysics Data System (ADS)

Fulton, Joseph M.

Due to the volume and power limitations of a small satellite, careful consideration must be taken while designing an attitude control system for 3-axis stabilization. Placing redundancy in the system proves difficult and utilizing power hungry, high accuracy, active actuators is not a viable option. Thus, it is customary to find dependable, passive actuators used in conjunction with small scale active control components. This document describes the application of Elastic Memory Composite materials in the construction of a flexible spacecraft appendage, such as a gravity gradient boom. Assumed modes methods are used with Finite Element Modeling information to obtain the equations of motion for the system while assuming free-free boundary conditions. A discussion is provided to illustrate how cantilever mode shapes are not always the best assumption when modeling small flexible spacecraft. A key point of interest is first resonant modes may be needed in the system design plant in spite of these modes being greater than one order of magnitude in frequency when compared to the crossover frequency of the controller. LQG/LTR optimal control techniques are implemented to compute attitude control gains while controller robustness considerations determine appropriate reduced order controllers and which flexible modes to include in the design model. Key satellite designer concerns in the areas of computer processor sizing, material uncertainty impacts on the system model, and system performance variations resulting from appendage length modifications are addressed.

Crack-free conditions in welding of glass by ultrashort laser pulse.

PubMed

Miyamoto, Isamu; Cvecek, Kristian; Schmidt, Michael

2013-06-17

The spatial distribution of the laser energy absorbed by nonlinear absorption process in bulk glass w(z) is determined and thermal cycles due to the successive ultrashort laser pulse (USLP) is simulated using w(z) based on the transient thermal conduction model. The thermal stress produced in internal melting of bulk glass by USLP is qualitatively analyzed based on a simple thermal stress model, and crack-free conditions are studied in glass having large coefficient of thermal expansion. In heating process, cracks are prevented when the laser pulse impinges into glass with temperatures higher than the softening temperature of glass. In cooling process, shrinkage stress is suppressed to prevent cracks, because the embedded molten pool produced by nonlinear absorption process behaves like an elastic body under the compressive stress field unlike the case of CW-laser welding where the molten pool having a free surface produced by linear absorption process is plastically deformed under the compressive stress field.

Stabilizing effect of resistivity towards ELM-free H-mode discharge in lithium-conditioned NSTX

DOE Office of Scientific and Technical Information (OSTI.GOV)

Banerjee, Debabrata; Zhu, Ping; Maingi, Rajesh

Linear stability analysis of the national spherical torus experiment (NSTX) Li-conditioned ELM-free H-mode equilibria is carried out in the context of the extended magneto-hydrodynamic (MHD) model in NIMROD. Our purpose is to investigate the physical cause behind edge localized mode (ELM) suppression in experiment after the Li-coating of the divertor and the first wall of the NSTX tokamak. Besides ideal MHD modeling, including finite-Larmor radius effect and two-fluid Hall and electron diamagnetic drift contributions, a non-ideal resistivity model is employed, taking into account the increase of Z eff after Li-conditioning in ELM-free H-mode. And unlike an earlier conclusion from anmore » eigenvalue code analysis of these equilibria, NIMROD results find that after reduced recycling from divertor plates, profile modification is necessary but insufficient to explain the mechanism behind complete ELMs suppression in ideal two-fluid MHD. After considering the higher plasma resistivity due to higher Z eff, the complete stabilization could be explained. Furthermore, a thorough analysis of both pre-lithium ELMy and with-lithium ELM-free cases using ideal and non-ideal MHD models is presented, after accurately including a vacuum-like cold halo region in NIMROD to investigate ELMs.« less

Stabilizing effect of resistivity towards ELM-free H-mode discharge in lithium-conditioned NSTX

DOE PAGES

Banerjee, Debabrata; Zhu, Ping; Maingi, Rajesh

2017-05-12

Linear stability analysis of the national spherical torus experiment (NSTX) Li-conditioned ELM-free H-mode equilibria is carried out in the context of the extended magneto-hydrodynamic (MHD) model in NIMROD. Our purpose is to investigate the physical cause behind edge localized mode (ELM) suppression in experiment after the Li-coating of the divertor and the first wall of the NSTX tokamak. Besides ideal MHD modeling, including finite-Larmor radius effect and two-fluid Hall and electron diamagnetic drift contributions, a non-ideal resistivity model is employed, taking into account the increase of Z eff after Li-conditioning in ELM-free H-mode. And unlike an earlier conclusion from anmore » eigenvalue code analysis of these equilibria, NIMROD results find that after reduced recycling from divertor plates, profile modification is necessary but insufficient to explain the mechanism behind complete ELMs suppression in ideal two-fluid MHD. After considering the higher plasma resistivity due to higher Z eff, the complete stabilization could be explained. Furthermore, a thorough analysis of both pre-lithium ELMy and with-lithium ELM-free cases using ideal and non-ideal MHD models is presented, after accurately including a vacuum-like cold halo region in NIMROD to investigate ELMs.« less

Improved Performance Analysis of Free Space Optics Communication Link under Rain Conditions using EDFA Pre-amplifier

NASA Astrophysics Data System (ADS)

Singh, Mehtab

2018-04-01

Free Space Optics (FSO) also known as Optical Wireless Communication (OWC) is a communication technology in which free space/air is used as the propagation medium and optical signals are used as the information carriers. One of the most crucial factors which degrade the performance of FSO link is the signal attenuation due to different atmospheric weather conditions such as haze, rain, storm, and fog. In this paper, an improved performance analysis of a 2.5 Gbps FSO link under rain conditions has been reported using Erbium-Doped Fiber Amplifier (EDFA) as a pre-amplifier. The results show that the maximum link distance for an FSO link under rain weather conditions with acceptable performance levels (Q ˜6 and BER ≤ 10-9 in the absence of EDFA pre-amplifier is 1,250 m which increases to 1,675 m with the use of EDFA pre-amplifier.

A comment on the paper 'A comparison between lesions found during meat inspection of finishing pigs raised under organic/free-range conditions and conventional indoor conditions' by Alban et al. 2015.

PubMed

Sørensen, Jan Tind

2016-01-01

This is a critical comment on a paper published in Porcine Health Management in 2015, presenting a comparison between lesions from meat inspection at one abattoir on slaughter pigs classified in to two different production forms: organic/free-range conditions and conventional indoor conditions. The conclusion made by the paper that 13 lesion types has a higher prevalence in organic/free-range pigs and 4 lesion types occurred less frequently in organic/free-range finishers compared to conventional finishers is correct except that 5 (instead of 4) lesion types occurred less frequently in organic/free-range finishers. However, these five types of lesions represent 74 % of all lesions recorded among conventional indoor, conventional free-range and organic pigs in one slaughter house from October 1 2012 to September 26 2013.

A Model-Free Diagnostic for Single-Peakedness of Item Responses Using Ordered Conditional Means

ERIC Educational Resources Information Center

Polak, Marike; De Rooij, Mark; Heiser, Willem J.

2012-01-01

In this article we propose a model-free diagnostic for single-peakedness (unimodality) of item responses. Presuming a unidimensional unfolding scale and a given item ordering, we approximate item response functions of all items based on ordered conditional means (OCM). The proposed OCM methodology is based on Thurstone & Chave's (1929) "criterion…

Defining treatment conditions for pulsed electric field pasteurization of apple juice.

PubMed

Saldaña, G; Puértolas, E; Monfort, S; Raso, J; Alvarez, I

2011-11-15

The influence of temperature and the presence of N(α)-lauroyl ethylester (ethyl lauroyl arginate, LAE) on the inactivation caused by continuous pulsed electric field treatments (PEF) in Escherichia coli O157:H7 suspended in apple juice have been investigated to define treatment conditions applicable at industrial scale that promote an equivalent safety level when compared with thermal processing. In the range of experimental conditions investigated (outlet temperature: 20-40 °C, electric field strength: 20-30 kV, treatment time: 5-125 μs) at outlet temperatures equal or lower than 55±1 °C, the inactivation of E. coli O157:H7 treated in apple juice ranged from 0.4 to 3.6 Log₁₀ cycles reduction and treated in apple juice supplemented with LAE (50 ppm) ranged from 0.9 to 6.7 Log₁₀ cycles reduction. An empirical mathematical model was developed to estimate the treatment time and total specific energy input to obtain 5 Log₁₀ cycles reduction in the population of E. coli O157:H7 suspended in apple juice supplemented with 50 ppm of LAE at different electric field strengths and inlet temperatures. Treatment conditions established for E. coli O157:H7 were validated with other PEF resistant Gram-positive (Listeria monocytogenes, and Staphylococcus aureus) and Gram-negative (Salmonella enterica serovar Typhimurium) strains. When the treatment was applied to the apple juice, a treatment of 25 kV/cm for 63 μs corresponding with an outlet temperature of 65 °C and input energy of 125 kJ/kg was required to achieve more than 5 Log₁₀ cycles in the four strains investigated. The addition of LAE reduced the treatment time required to obtain an equivalent inactivation (>5 Log₁₀ cycles) in the four microorganisms to 38.4 μs, the outlet temperature to 55 °C, and the input energy to 83.2 kJ/kg. Copyright © 2011 Elsevier B.V. All rights reserved.

Developmental Competence of Buffalo (Bubalus bubalis) Pluripotent Embryonic Stem Cells Over Different Homologous Feeder Layers and the Comparative Evaluation with Various Extracellular Matrices.

PubMed

Sharma, Manjinder; Dubey, Pawan K; Kumar, Rajesh; Nath, Amar; Kumar, G Sai; Sharma, G Taru

2013-05-01

Use of somatic cells as a feeder layer to maintain the embryonic stem cells (ESCs) in undifferentiated state limits the stem cell research design, since experimental data may result from a combined ESCs and feeder cell response to various stimuli. Therefore, present study was designed to evaluate the developmental competence of the buffalo ESCs over different homogenous feeders and compare with various extracellular matrices using different concentrations of LIF. Inner cell masses (ICMs) of in vitro hatched blastocysts were cultured onto homologous feeders viz. fetal fibroblast, granulosa and oviductal cell feeder layers and synthetic matrices viz. fibronectin, collagen type I and matrigel in culture medium. Developmental efficiency was found higher for ESCs cultured on fetal fibroblast and granulosa layers (83.33%) followed by fibronectin (77.78%) at 30 ng LIF. Oviductal feeder was found to be the least efficient feeder showing only 11.11% undifferentiated primary ESC colonies at 30 ng LIF. However, neither feeder layer nor synthetic matrix could support the development of primary colonies at 10 ng LIF. Expression of SSEA- 4, TRA-1-60 and Oct-4 were found positive in ESC colonies from all the feeders and synthetic matrices with 20 ng and 30 ng LIF. Fetal fibroblast and granulosa cell while, amongst synthetic matrices, fibronectin were found to be equally efficient to support the growth and maintenance of ESCs pluripotency with 30 ng LIF. This well-defined culture conditions may provide an animal model for culturing human embryonic stem cells in the xeno-free or feeder-free conditions for future clinical applications.

Practical methods for handling human periodontal ligament stem cells in serum-free and serum-containing culture conditions under hypoxia: implications for regenerative medicine.

PubMed

Murabayashi, Dai; Mochizuki, Mai; Tamaki, Yuichi; Nakahara, Taka

2017-07-01

Stem cell-based therapies depend on the reliable expansion of patient-derived mesenchymal stem cells (MSCs) in vitro. The supplementation of cell culture media with serum is associated with several risks; accordingly, serum-free media are commercially available for cell culture. Furthermore, hypoxia is known to accelerate the expansion of MSCs. The present study aimed to characterize the properties of periodontal ligament-derived MSCs (PDLSCs) cultivated in serum-free and serum-containing media, under hypoxic and normoxic conditions. Cell growth, gene and protein expression, cytodifferentiation potential, genomic stability, cytotoxic response, and in vivo hard tissue generation of PDLSCs were examined. Our findings indicated that cultivation in serum-free medium does not affect the MSC phenotype or chromosomal stability of PDLSCs. PDLSCs expanded in serum-free medium exhibited more active growth than in fetal bovine serum-containing medium. We found that hypoxia does not alter the cell growth of PDLSCs under serum-free conditions, but inhibits their osteogenic and adipogenic cytodifferentiation while enabling maintenance of their multidifferentiation potential regardless of the presence of serum. PDLSCs expanded in serum-free medium were found to retain common MSC characteristics, including the capacity for hard tissue formation in vivo. However, PDLSCs cultured in serum-free culture conditions were more susceptible to damage following exposure to extrinsic cytotoxic stimuli than those cultured in medium supplemented with serum, suggesting that serum-free culture conditions do not exert protective effects against cytotoxicity on PDLSC cultures. The present work provides a comparative evaluation of cell culture in serum-free and serum-containing media, under hypoxic and normoxic conditions, for applications in regenerative medicine.

Stabilizing Effect of Resistivity towards ELM-free H-mode Discharge in Lithium-conditioned NSTX

NASA Astrophysics Data System (ADS)

Banerjee, Debabrata; Zhu, Ping; Maingi, Rajesh

2016-10-01

The stabilizing effect of edge resistivity on the edge localized modes (ELMs) has been recently recovered through analyzing NSTX experimental profiles of Lithium-conditioned ELM-free H-mode discharge. Comparative studies of ELM-free and a reference NSTX ELMy-H mode equilibriums have been performed using both resistive and 2-fluid MHD models implemented in the initial value extended MHD code NIMROD. Our results indicate that in addition to the pedestal profile refinement in electron pressure, the inclusion of enhanced resistivity due to the increase in the effective electric charge number Zeff, which is observed after Lithium-conditioning in experiment, is further required to account for the full stabilization of the low- n edge localized modes. Such a stabilization from the enhanced edge resistivity only becomes effective when the two-fluid diamagnetic and finite-Larmor-radius (FLR) effects are considered in the MHD model. Supported by the National Magnetic Confinement Fusion Program of China under Grant Nos. 2014GB124002 and 2015GB101004, the 100 Talent Program and the President International Fellowship Initiative of the Chinese Academy of Sciences.

Simulation of VSPT Experimental Cascade Under High and Low Free-Stream Turbulence Conditions

NASA Technical Reports Server (NTRS)

Ameri, Ali A.; Giel, Paul W.; Flegel, Ashlie B.

2014-01-01

Variable-Speed Power Turbines (VSPT) for rotorcraft applications operate at low Reynolds number and over a wide range in incidence associated with shaft speed change. A comprehensive linear cascade data set obtained includes the effects of Reynolds number, free-stream turbulence and incidence is available and this paper concerns itself with the presentation and numerical simulation of conditions resulting in a selected set of those data. As such, post-dictions of blade pressure loading, total-pressure loss and exit flow angles under conditions of high and low turbulence intensity for a single Reynolds number are presented. Analyses are performed with the three-equation turbulence models of Walters-Leylek and Walters and Cokljat. Transition, loading, total-pressure loss and exit angle variations are presented and comparisons are made with experimental data as available. It is concluded that at the low freestream turbulence conditions the Walters-Cokljat model is better suited to predictions while for high freestream conditions the two models generate similar predications that are generally satisfactory.

Simulation of VSPT Experimental Cascade Under High and Low Free-Stream Turbulence Conditions

NASA Technical Reports Server (NTRS)

Ameri, Ali A.; Giel, Paul W.; Flegel, Ashlie B.

2015-01-01

Variable-Speed Power Turbines (VSPT) for rotorcraft applications operate at low Reynolds number and over a wide range in incidence associated with shaft speed change. A comprehensive linear cascade data set obtained includes the effects of Reynolds number, free-stream turbulence and incidence is available and this paper concerns itself with the presentation and numerical simulation of conditions resulting in a selected set of those data. As such, post-dictions of blade pressure loading, total-pressure loss and exit flow angles under conditions of high and low turbulence intensity for a single Reynolds number are presented. Analyses are performed with the three-equation turbulence models of Walters- Leylek and Walters and Cokljat. Transition, loading, total-pressure loss and exit angle variations are presented and comparisons are made with experimental data as available. It is concluded that at the low freestream turbulence conditions the Walters-Cokljat model is better suited to predictions while for high freestream conditions the two models generate similar predications that are generally satisfactory.

Pulsed electric fields for pasteurization: defining processing conditions

USDA-ARS?s Scientific Manuscript database

Application of pulsed electric fields (PEF) technology in food pasteurization has been extensively studied. Optimal PEF treatment conditions for maximum microbial inactivation depend on multiple factors including PEF processing conditions, production parameters and product properties. In order for...

Disclosure of the differences of Mesorhizobium loti under the free-living and symbiotic conditions by comparative proteome analysis without bacteroid isolation.

PubMed

Tatsukami, Yohei; Nambu, Mami; Morisaka, Hironobu; Kuroda, Kouichi; Ueda, Mitsuyoshi

2013-07-31

Rhizobia are symbiotic nitrogen-fixing soil bacteria that show a symbiotic relationship with their host legume. Rhizobia have 2 different physiological conditions: a free-living condition in soil, and a symbiotic nitrogen-fixing condition in the nodule. The lifestyle of rhizobia remains largely unknown, although genome and transcriptome analyses have been carried out. To clarify the lifestyle of bacteria, proteome analysis is necessary because the protein profile directly reflects in vivo reactions of the organisms. In proteome analysis, high separation performance is required to analyze complex biological samples. Therefore, we used a liquid chromatography-tandem mass spectrometry system, equipped with a long monolithic silica capillary column, which is superior to conventional columns. In this study, we compared the protein profile of Mesorhizobium loti MAFF303099 under free-living condition to that of symbiotic conditions by using small amounts of crude extracts. We identified 1,533 and 847 proteins for M. loti under free-living and symbiotic conditions, respectively. Pathway analysis by Kyoto Encyclopedia of Genes and Genomes (KEGG) revealed that many of the enzymes involved in the central carbon metabolic pathway were commonly detected under both conditions. The proteins encoded in the symbiosis island, the transmissible chromosomal region that includes the genes that are highly upregulated under the symbiotic condition, were uniquely detected under the symbiotic condition. The features of the symbiotic condition that have been reported by transcriptome analysis were confirmed at the protein level by proteome analysis. In addition, the genes of the proteins involved in cell surface structure were repressed under the symbiotic nitrogen-fixing condition. Furthermore, farnesyl pyrophosphate (FPP) was found to be biosynthesized only in rhizobia under the symbiotic condition. The obtained protein profile appeared to reflect the difference in phenotypes under the

Relations between water physico-chemistry and benthic algal communities in a northern Canadian watershed: defining reference conditions using multiple descriptors of community structure.

PubMed

Thomas, Kathryn E; Hall, Roland I; Scrimgeour, Garry J

2015-09-01

Defining reference conditions is central to identifying environmental effects of anthropogenic activities. Using a watershed approach, we quantified reference conditions for benthic algal communities and their relations to physico-chemical conditions in rivers in the South Nahanni River watershed, NWT, Canada, in 2008 and 2009. We also compared the ability of three descriptors that vary in terms of analytical costs to define algal community structure based on relative abundances of (i) all algal taxa, (ii) only diatom taxa, and (iii) photosynthetic pigments. Ordination analyses showed that variance in algal community structure was strongly related to gradients in environmental variables describing water physico-chemistry, stream habitats, and sub-watershed structure. Water physico-chemistry and local watershed-scale descriptors differed significantly between algal communities from sites in the Selwyn Mountain ecoregion compared to sites in the Nahanni-Hyland ecoregions. Distinct differences in algal community types between ecoregions were apparent irrespective of whether algal community structure was defined using all algal taxa, diatom taxa, or photosynthetic pigments. Two algal community types were highly predictable using environmental variables, a core consideration in the development of Reference Condition Approach (RCA) models. These results suggest that assessments of environmental impacts could be completed using RCA models for each ecoregion. We suggest that use of algal pigments, a high through-put analysis, is a promising alternative compared to more labor-intensive and costly taxonomic approaches for defining algal community structure.

Defining Chlorophyll-a Reference Conditions in European Lakes

PubMed Central

Alves, Maria Helena; Argillier, Christine; van den Berg, Marcel; Buzzi, Fabio; Hoehn, Eberhard; de Hoyos, Caridad; Karottki, Ivan; Laplace-Treyture, Christophe; Solheim, Anne Lyche; Ortiz-Casas, José; Ott, Ingmar; Phillips, Geoff; Pilke, Ansa; Pádua, João; Remec-Rekar, Spela; Riedmüller, Ursula; Schaumburg, Jochen; Serrano, Maria Luisa; Soszka, Hanna; Tierney, Deirdre; Urbanič, Gorazd; Wolfram, Georg

2010-01-01

The concept of “reference conditions” describes the benchmark against which current conditions are compared when assessing the status of water bodies. In this paper we focus on the establishment of reference conditions for European lakes according to a phytoplankton biomass indicator—the concentration of chlorophyll-a. A mostly spatial approach (selection of existing lakes with no or minor human impact) was used to set the reference conditions for chlorophyll-a values, supplemented by historical data, paleolimnological investigations and modelling. The work resulted in definition of reference conditions and the boundary between “high” and “good” status for 15 main lake types and five ecoregions of Europe: Alpine, Atlantic, Central/Baltic, Mediterranean, and Northern. Additionally, empirical models were developed for estimating site-specific reference chlorophyll-a concentrations from a set of potential predictor variables. The results were recently formulated into the EU legislation, marking the first attempt in international water policy to move from chemical quality standards to ecological quality targets. PMID:20401659

Towards Defining Nutrient Conditions Encountered by the Rice Blast Fungus during Host Infection

PubMed Central

Wilson, Richard A.; Fernandez, Jessie; Quispe, Cristian F.; Gradnigo, Julien; Seng, Anya; Moriyama, Etsuko; Wright, Janet D.

2012-01-01

Fungal diseases cause enormous crop losses, but defining the nutrient conditions encountered by the pathogen remains elusive. Here, we generated a mutant strain of the devastating rice pathogen Magnaporthe oryzae impaired for de novo methionine biosynthesis. The resulting methionine-requiring strain grew strongly on synthetic minimal media supplemented with methionine, aspartate or complex mixtures of partially digested proteins, but could not establish disease in rice leaves. Live-cell-imaging showed the mutant could produce normal appressoria and enter host cells but failed to develop, indicating the availability or accessibility of aspartate and methionine is limited in the plant. This is the first report to demonstrate the utility of combining biochemical genetics, plate growth tests and live-cell-imaging to indicate what nutrients might not be readily available to the fungal pathogen in rice host cells. PMID:23071797

Antenna gain of actively compensated free-space optical communication systems under strong turbulence conditions.

PubMed

Juarez, Juan C; Brown, David M; Young, David W

2014-05-19

Current Strehl ratio models for actively compensated free-space optical communications terminals do not accurately predict system performance under strong turbulence conditions as they are based on weak turbulence theory. For evaluation of compensated systems, we present an approach for simulating the Strehl ratio with both low-order (tip/tilt) and higher-order (adaptive optics) correction. Our simulation results are then compared to the published models and their range of turbulence validity is assessed. Finally, we propose a new Strehl ratio model and antenna gain equation that are valid for general turbulence conditions independent of the degree of compensation.

Rifampin modulation of xeno- and endobiotic conjugating enzyme mRNA expression and associated microRNAs in human hepatocytes.

PubMed

Gufford, Brandon T; Robarge, Jason D; Eadon, Michael T; Gao, Hongyu; Lin, Hai; Liu, Yunlong; Desta, Zeruesenay; Skaar, Todd C

2018-04-01

Rifampin is a pleiotropic inducer of multiple drug metabolizing enzymes and transporters. This work utilized a global approach to evaluate rifampin effects on conjugating enzyme gene expression with relevance to human xeno- and endo-biotic metabolism. Primary human hepatocytes from 7 subjects were treated with rifampin (10 μmol/L, 24 hours). Standard methods for RNA-seq library construction, EZBead preparation, and NextGen sequencing were used to measure UDP-glucuronosyl transferase UGT, sulfonyltransferase SULT, N acetyltransferase NAT, and glutathione-S-transferase GST mRNA expression compared to vehicle control (0.01% MeOH). Rifampin-induced (>1.25-fold) mRNA expression of 13 clinically important phase II drug metabolizing genes and repressed (>1.25-fold) the expression of 3 genes ( P  <   .05). Rifampin-induced miRNA expression changes correlated with mRNA changes and miRNAs were identified that may modulate conjugating enzyme expression. NAT2 gene expression was most strongly repressed (1.3-fold) by rifampin while UGT1A4 and UGT1A1 genes were most strongly induced (7.9- and 4.8-fold, respectively). Physiologically based pharmacokinetic modeling (PBPK) was used to simulate the clinical consequences of rifampin induction of CYP3A4- and UGT1A4-mediated midazolam metabolism. Simulations evaluating isolated UGT1A4 induction predicted increased midazolam N-glucuronide exposure (~4-fold) with minimal reductions in parent midazolam exposure (~10%). Simulations accounting for simultaneous induction of both CYP3A4 and UGT1A4 predicted a ~10-fold decrease in parent midazolam exposure with only a ~2-fold decrease in midazolam N-glucuronide metabolite exposure. These data reveal differential effects of rifampin on the human conjugating enzyme transcriptome and potential associations with miRNAs that form the basis for future mechanistic studies to elucidate the interplay of conjugating enzyme regulatory elements.

Enhancement of acetylcholine-induced desensitization of guinea-pig ileal longitudinal muscle in Ca2+-free conditions.

PubMed

Horio, S; Nagare, T; Moritoki, H

1999-10-01

1. To determine the role of cellular Ca2+ in desensitization, acetylcholine(ACh)-induced desensitization was studied under Ca2+-free condition in guinea-pig ileal longitudinal muscle. 2. Pretreatment of the tissue with 10(-4) M ACh (desensitizing treatment) in normal Tyrode solution caused desensitization of the responses both to ACh and histamine. The desensitizing treatment performed in Ca2+-free solution enhanced desensitization of the responses to ACh and histamine significantly. 3. The desensitizing treatment with ACh caused suppression of the responses to high K+ (tonic component) and Bay K 8644. The desensitizing treatment performed in Ca2+-free solution potentiated the suppression of the responses to high K+ and Bay K 8644 significantly. 4. ACh-induced desensitization was enhanced significantly in the presence of a protein kinase C inhibitor, 1-(5-isoquinolinesulfonyl)-2-methylpiperazine(H-7, 10(-4) M) to a similar extent as desensitization obtained under Ca2+-free condition, but not in the presence of a non-specific and less potent kinase inhibitor, N-(2-guanidinoethyl)-5-isoquinolinesulfonamide hydrochloride (HA1004, 10(-4) M). 5. These results suggested that voltage-gated Ca2+ channels were involved in ACh-induced desensitization and that intracellular Ca2+, which was increased during the stimulation with ACh, inhibited desensitization through the activation of protein kinase C. This kinase could have activated or protected Ca2+ channels during the desensitization process to reduce desensitization.

Interfacial free energy and stiffness of aluminum during rapid solidification

DOE Office of Scientific and Technical Information (OSTI.GOV)

Brown, Nicholas T.; Martinez, Enrique; Qu, Jianmin

Using molecular dynamics simulations and the capillary fluctuation method, we have calculated the anisotropic crystal-melt interfacial free energy and stiffness of aluminum in a rapid solidification system where a temperature gradient is applied to enforce thermal non-equilibrium. To calculate these material properties, the standard capillary fluctuation method typically used for systems in equilibrium has been modified to incorporate a second-order Taylor expansion of the interfacial free energy term. The result is a robust method for calculating interfacial energy, stiffness and anisotropy as a function of temperature gradient using the fluctuations in the defined interface height. This work includes the calculationmore » of interface characteristics for temperature gradients ranging from 11 to 34 K/nm. The captured results are compared to a thermal equilibrium case using the same model and simulation technique with a zero gradient definition. We define the temperature gradient as the change in temperature over height perpendicular to the crystal-melt interface. The gradients are applied in MD simulations using defined thermostat regions on a stable solid-liquid interface initially in thermal equilibrium. The results of this work show that the interfacial stiffness and free energy for aluminum are dependent on the magnitude of the temperature gradient, however the anisotropic parameters remain independent of the non-equilibrium conditions applied in this analysis. As a result, the relationships of the interfacial free energy/stiffness are determined to be linearly related to the thermal gradient, and can be interpolated to find material characteristics at additional temperature gradients.« less

Interfacial free energy and stiffness of aluminum during rapid solidification

DOE PAGES

Brown, Nicholas T.; Martinez, Enrique; Qu, Jianmin

2017-05-01

Using molecular dynamics simulations and the capillary fluctuation method, we have calculated the anisotropic crystal-melt interfacial free energy and stiffness of aluminum in a rapid solidification system where a temperature gradient is applied to enforce thermal non-equilibrium. To calculate these material properties, the standard capillary fluctuation method typically used for systems in equilibrium has been modified to incorporate a second-order Taylor expansion of the interfacial free energy term. The result is a robust method for calculating interfacial energy, stiffness and anisotropy as a function of temperature gradient using the fluctuations in the defined interface height. This work includes the calculationmore » of interface characteristics for temperature gradients ranging from 11 to 34 K/nm. The captured results are compared to a thermal equilibrium case using the same model and simulation technique with a zero gradient definition. We define the temperature gradient as the change in temperature over height perpendicular to the crystal-melt interface. The gradients are applied in MD simulations using defined thermostat regions on a stable solid-liquid interface initially in thermal equilibrium. The results of this work show that the interfacial stiffness and free energy for aluminum are dependent on the magnitude of the temperature gradient, however the anisotropic parameters remain independent of the non-equilibrium conditions applied in this analysis. As a result, the relationships of the interfacial free energy/stiffness are determined to be linearly related to the thermal gradient, and can be interpolated to find material characteristics at additional temperature gradients.« less

Stepwise differentiation of pluripotent stem cells into osteoblasts using four small molecules under serum-free and feeder-free conditions.

PubMed

Kanke, Kosuke; Masaki, Hideki; Saito, Taku; Komiyama, Yuske; Hojo, Hironori; Nakauchi, Hiromitsu; Lichtler, Alexander C; Takato, Tsuyoshi; Chung, Ung-Il; Ohba, Shinsuke

2014-06-03

Pluripotent stem cells are a promising tool for mechanistic studies of tissue development, drug screening, and cell-based therapies. Here, we report an effective and mass-producing strategy for the stepwise differentiation of mouse embryonic stem cells (mESCs) and mouse and human induced pluripotent stem cells (miPSCs and hiPSCs, respectively) into osteoblasts using four small molecules (CHIR99021 [CHIR], cyclopamine [Cyc], smoothened agonist [SAG], and a helioxanthin-derivative 4-(4-methoxyphenyl)pyrido[4',3':4,5]thieno[2,3-b]pyridine-2-carboxamide [TH]) under serum-free and feeder-free conditions. The strategy, which consists of mesoderm induction, osteoblast induction, and osteoblast maturation phases, significantly induced expressions of osteoblast-related genes and proteins in mESCs, miPSCs, and hiPSCs. In addition, when mESCs defective in runt-related transcription factor 2 (Runx2), a master regulator of osteogenesis, were cultured by the strategy, they molecularly recapitulated osteoblast phenotypes of Runx2 null mice. The present strategy will be a platform for biological and pathological studies of osteoblast development, screening of bone-augmentation drugs, and skeletal regeneration.

Probing Chirality in Superfluid 3He-A: Free surface as an ideal boundary condition

NASA Astrophysics Data System (ADS)

Kono, Kimitoshi

2013-03-01

Superfluid 3He is known as a typical topological superfluid. A recent theoretical investigation suggests Majorana surface states at the free surface of superfluid 3He-B phase. On the other hand, superfluid 3He-A is known as a chiral superfluid. The scattering of quasiparticle from small object is predicted to be skew with respect to an anisotropy axis. We have developed an experimental technique to study transport properties of ions under the free surface of superfluid 3He. By using this technique, we can investigate interaction between elementary excitations in superfluid 3He and small objects under well-controlled conditions. For example, in 3He-B interaction with Majorana surface states, although no interaction is expected, will be investigated, whereas in 3He-A skew scattering of quasiparticle from electron bubbles will be probed. In this paper, we present the recent results of transport properties of electron bubbles trapped below the free surface of superfluid 3He. In particular, experimental evidences of the skew scattering and chirality of superfluid 3He-A will be presented. The skew scattering of quasiparticle in 3He-A from electron bubble results in a bubble transport analogous to the Hall effect, where the anisotropy vector of 3He-A behaves as if it was a magnetic field in the Hall effect. Under experimental conditions, the effect is observed as an analogue of edge magnetoplasmon effect. After the analysis of data, we obtained a reasonable qualitative agreement with the theory. This work is done in collaboration with Hiroki Ikegami.

β-cell serotonin production is associated with female sex, old age, and diabetes-free condition.

PubMed

Kim, Yeong Gi; Moon, Joon Ho; Kim, Kyuho; Kim, Hyeongseok; Kim, Juok; Jeong, Ji-Seon; Lee, Junguee; Kang, Shinae; Park, Joon Seong; Kim, Hail

2017-11-25

Serotonin is known to be present in pancreatic β-cells and to play several physiological roles, including insulin secretion, β-cell proliferation, and paracrine inhibition of α-cells. However, the serotonin production of different cell lines and islets has not been compared based on age, sex, and diabetes related conditions. Here, we directly compared the serotonin concentrations in βTC and MIN6 cell lines, as well as in islets from mice using ultra-performance liquid chromatography tandem mass spectrometry. The average serotonin concentration was 5-10 ng/mg protein in the islets of male and non-pregnant female mice. The serotonin level was higher in females than males at 8 weeks, although there was no difference at 1 year. Furthermore, we observed serotonin by immunofluorescence staining in the pancreatic tissues of mice and human. Serotonin was detected by immunofluorescence staining in a portion of β-cells from islets of old female mice, but not of male or young female mice. A similar pattern was observed in human pancreas as well. In humans, serotonin production in β-cells was associated with a diabetes-free condition. Thus, serotonin production in β-cells was associated with old age, female sex, and diabetes-free condition. Copyright © 2017 Elsevier Inc. All rights reserved.

Defined Medium Conditions for the Induction and Expansion of Human Pluripotent Stem Cell-Derived Retinal Pigment Epithelium.

PubMed

Lidgerwood, Grace E; Lim, Shiang Y; Crombie, Duncan E; Ali, Ray; Gill, Katherine P; Hernández, Damián; Kie, Josh; Conquest, Alison; Waugh, Hayley S; Wong, Raymond C B; Liang, Helena H; Hewitt, Alex W; Davidson, Kathryn C; Pébay, Alice

2016-04-01

We demonstrate that a combination of Noggin, Dickkopf-1, Insulin Growth Factor 1 and basic Fibroblast Growth Factor, promotes the differentiation of human pluripotent stem cells into retinal pigment epithelium (RPE) cells. We describe an efficient one-step approach that allows the generation of RPE cells from both human embryonic stem cells and human induced pluripotent stem cells within 40-60 days without the need for manual excision, floating aggregates or imbedded cysts. Compared to methods that rely on spontaneous differentiation, our protocol results in faster differentiation into RPE cells. This pro-retinal culture medium promotes the growth of functional RPE cells that exhibit key characteristics of the RPE including pigmentation, polygonal morphology, expression of mature RPE markers, electrophysiological membrane potential and the ability to phagocytose photoreceptor outer segments. This protocol can be adapted for feeder, feeder-free and serum-free conditions. This method thereby provides a rapid and simplified production of RPE cells for downstream applications such as disease modelling and drug screening.

Predicting solvation free energies and thermodynamics in polar solvents and mixtures using a solvation-layer interface condition

PubMed Central

Goossens, Spencer; Mehdizadeh Rahimi, Ali

2017-01-01

We demonstrate that with two small modifications, the popular dielectric continuum model is capable of predicting, with high accuracy, ion solvation thermodynamics (Gibbs free energies, entropies, and heat capacities) in numerous polar solvents. We are also able to predict ion solvation free energies in water–co-solvent mixtures over available concentration series. The first modification to the classical dielectric Poisson model is a perturbation of the macroscopic dielectric-flux interface condition at the solute–solvent interface: we add a nonlinear function of the local electric field, giving what we have called a solvation-layer interface condition (SLIC). The second modification is including the microscopic interface potential (static potential) in our model. We show that the resulting model exhibits high accuracy without the need for fitting solute atom radii in a state-dependent fashion. Compared to experimental results in nine water–co-solvent mixtures, SLIC predicts transfer free energies to within 2.5 kJ/mol. The co-solvents include both protic and aprotic species, as well as biologically relevant denaturants such as urea and dimethylformamide. Furthermore, our results indicate that the interface potential is essential to reproduce entropies and heat capacities. These and previous tests of the SLIC model indicate that it is a promising dielectric continuum model for accurate predictions in a wide range of conditions.

Predicting solvation free energies and thermodynamics in polar solvents and mixtures using a solvation-layer interface condition

NASA Astrophysics Data System (ADS)

Molavi Tabrizi, Amirhossein; Goossens, Spencer; Mehdizadeh Rahimi, Ali; Knepley, Matthew; Bardhan, Jaydeep P.

2017-03-01

We demonstrate that with two small modifications, the popular dielectric continuum model is capable of predicting, with high accuracy, ion solvation thermodynamics (Gibbs free energies, entropies, and heat capacities) in numerous polar solvents. We are also able to predict ion solvation free energies in water-co-solvent mixtures over available concentration series. The first modification to the classical dielectric Poisson model is a perturbation of the macroscopic dielectric-flux interface condition at the solute-solvent interface: we add a nonlinear function of the local electric field, giving what we have called a solvation-layer interface condition (SLIC). The second modification is including the microscopic interface potential (static potential) in our model. We show that the resulting model exhibits high accuracy without the need for fitting solute atom radii in a state-dependent fashion. Compared to experimental results in nine water-co-solvent mixtures, SLIC predicts transfer free energies to within 2.5 kJ/mol. The co-solvents include both protic and aprotic species, as well as biologically relevant denaturants such as urea and dimethylformamide. Furthermore, our results indicate that the interface potential is essential to reproduce entropies and heat capacities. These and previous tests of the SLIC model indicate that it is a promising dielectric continuum model for accurate predictions in a wide range of conditions.

Free-jet Testing of a REST Scramjet at Off-Design Conditions

NASA Technical Reports Server (NTRS)

Smart, Michael K.; Ruf, Edward G.

2006-01-01

Scramjet flowpaths employing elliptical combustors have the potential to improve structural efficiency and performance relative to those using planar geometries. NASA Langley has developed a scramjet flowpath integrated into a lifting body vehicle, while transitioning from a rectangular capture area to both an elliptical throat and combustor. This Rectangular-to-Elliptical Shape Transition (REST) scramjet, has a design point of Mach 7.1, and is intended to operate with fixed-geometry between Mach 4.5 and 8.0. This paper describes initial free-jet testing of the heat-sink REST scramjet engine model at conditions simulating Mach 5.3 flight. Combustion of gaseous hydrogen fuel at equivalence ratios between 0.5 and 1.5 generated robust performance after ignition with a silane-hydrogen pilot. Facility model interactions were experienced for fuel equivalence ratios above 1.1, yet despite this, the flowpath was not unstarted by fuel addition at the Mach 5.3 test condition. Combustion tests at reduced stagnation enthalpy indicated that the engine self-started following termination of the fuel injection. Engine data is presented for the largest fuel equivalence ratio tested without facility interaction. These results indicate that this class of three-dimensional scramjet engine operates successfully at off-design conditions.

Condition assessment of corroded steel rebar in free space using synthetic aperture radar images

NASA Astrophysics Data System (ADS)

Ingemi, Christopher M.; Owusu Twumasi, Jones; Litt, Swinderjit; Yu, Tzuyang

2017-04-01

Synthetic aperture radar (SAR) imaging of construction materials offers civil engineers an opportunity to better assess the condition of aging civil infrastructures such as reinforced concrete (RC) structures. Corrosion of steel rebar in RC structures is a major problem responsible for their premature failure and unexpected collapse. In this paper, SAR imaging is applied to the quantitative assessment of corroded steel rebar in free space as the first step toward the use of SAR imaging for subsurface sensing of aging RC structures. A 10 GHz stripmap SAR system was used inside an anechoic chamber. The bandwidth of the radar system was 1.5 GHz. Steel rebar specimens were artificially corroded to different levels by regularly applying a mist of 5% NaCl solution for different durations of time in order to simulate the condition of natural corrosion. Two sizes (No. 3 and No. 4) of steel rebar were used in this research. Different orientations of steel rebar were considered. Corrosion level was determined by measuring the mass loss of corroded steel rebar specimens. From our results, feasibility of SAR images for the condition assessment of corroded steel rebar was experimentally demonstrated. It was found that the presence of surface rust on corroded steel rebar reduces the amplitude in SAR images. The SAR image of corroded steel rebar also exhibited a distribution of SAR amplitudes different from the one of intact steel rebar. In addition, it was also found that there is an optimal range for the condition assessment of corroded steel rebar in free space. In our experiment, the optimal range was determined to be 30.4 cm.

Disclosure of the differences of Mesorhizobium loti under the free-living and symbiotic conditions by comparative proteome analysis without bacteroid isolation

PubMed Central

2013-01-01

Background Rhizobia are symbiotic nitrogen-fixing soil bacteria that show a symbiotic relationship with their host legume. Rhizobia have 2 different physiological conditions: a free-living condition in soil, and a symbiotic nitrogen-fixing condition in the nodule. The lifestyle of rhizobia remains largely unknown, although genome and transcriptome analyses have been carried out. To clarify the lifestyle of bacteria, proteome analysis is necessary because the protein profile directly reflects in vivo reactions of the organisms. In proteome analysis, high separation performance is required to analyze complex biological samples. Therefore, we used a liquid chromatography-tandem mass spectrometry system, equipped with a long monolithic silica capillary column, which is superior to conventional columns. In this study, we compared the protein profile of Mesorhizobium loti MAFF303099 under free-living condition to that of symbiotic conditions by using small amounts of crude extracts. Result We identified 1,533 and 847 proteins for M. loti under free-living and symbiotic conditions, respectively. Pathway analysis by Kyoto Encyclopedia of Genes and Genomes (KEGG) revealed that many of the enzymes involved in the central carbon metabolic pathway were commonly detected under both conditions. The proteins encoded in the symbiosis island, the transmissible chromosomal region that includes the genes that are highly upregulated under the symbiotic condition, were uniquely detected under the symbiotic condition. The features of the symbiotic condition that have been reported by transcriptome analysis were confirmed at the protein level by proteome analysis. In addition, the genes of the proteins involved in cell surface structure were repressed under the symbiotic nitrogen-fixing condition. Furthermore, farnesyl pyrophosphate (FPP) was found to be biosynthesized only in rhizobia under the symbiotic condition. Conclusion The obtained protein profile appeared to reflect the

Rearing the Fruit Fly Drosophila melanogaster Under Axenic and Gnotobiotic Conditions.

PubMed

Koyle, Melinda L; Veloz, Madeline; Judd, Alec M; Wong, Adam C-N; Newell, Peter D; Douglas, Angela E; Chaston, John M

2016-07-30

The influence of microbes on myriad animal traits and behaviors has been increasingly recognized in recent years. The fruit fly Drosophila melanogaster is a model for understanding microbial interactions with animal hosts, facilitated by approaches to rear large sample sizes of Drosophila under microorganism-free (axenic) conditions, or with defined microbial communities (gnotobiotic). This work outlines a method for collection of Drosophila embryos, hypochlorite dechorionation and sterilization, and transfer to sterile diet. Sterilized embryos are transferred to sterile diet in 50 ml centrifuge tubes, and developing larvae and adults remain free of any exogenous microbes until the vials are opened. Alternatively, flies with a defined microbiota can be reared by inoculating sterile diet and embryos with microbial species of interest. We describe the introduction of 4 bacterial species to establish a representative gnotobiotic microbiota in Drosophila. Finally, we describe approaches for confirming bacterial community composition, including testing if axenic Drosophila remain bacteria-free into adulthood.

Free and cued memory in relation to biomarker-defined abnormalities in clinically normal older adults and those at risk for Alzheimer's disease.

PubMed

Papp, Kathryn V; Amariglio, Rebecca E; Mormino, Elizabeth C; Hedden, Trey; Dekhytar, Maria; Johnson, Keith A; Sperling, Reisa A; Rentz, Dorene M

2015-07-01

Furthering our understanding of the relationship between amyloidosis (Aβ), neurodegeneration (ND), and cognition is imperative for early identification and early intervention of Alzheimer's disease (AD). However, the subtle cognitive decline differentially associated with each biomarker-defined stage of preclinical AD has yet to be fully characterized. Recent work indicates that different components of memory performance (free and cued recall) may be differentially specific to memory decline in prodromal AD. We sought to examine the relationship between free and cued recall paradigms, in addition to global composites of memory, executive functioning, and processing speed in relation to stages of preclinical AD. A total of 260 clinically normal (CN) older adults (CDR=0) from the Harvard Aging Brain study were grouped according to preclinical AD stages including Stage 0 (Aβ-/ND-), Stage 1 (Aβ+/ND-), Stage 2 (Aβ+/ND+), and suspected non-Alzheimer's associated pathology (SNAP; Aβ-/ND+). General linear models controlling for age, sex, and education were used to assess for stage-based performance differences on cognitive composites of executive functioning, processing speed, and memory in addition to free and cued delayed recall on the Selective Reminding Test (SRT) and Memory Capacity Test (MCT). Global memory performance differed between preclinical stages with Stage 2 performing worse compared with Stage 0. When examining free and cued paradigms by memory test, only the MCT (and not the SRT) revealed group differences. More specifically, Stage 1 was associated with decrements in free recall compared with Stage 0 while Stage 2 was associated with decrements in both free and cued recall. There was a trend for the SNAP group to perform worse on free recall compared with Stage 0. Finally, there was no association between preclinical stage and global composites of executive functioning or processing speed. Clinically normal older adults with underlying evidence of

Prognosis and Conditional Disease-Free Survival Among Patients With Ovarian Cancer

PubMed Central

Kurta, Michelle L.; Edwards, Robert P.; Moysich, Kirsten B.; McDonough, Kathleen; Bertolet, Marnie; Weissfeld, Joel L.; Catov, Janet M.; Modugno, Francesmary; Bunker, Clareann H.; Ness, Roberta B.; Diergaarde, Brenda

2014-01-01

Purpose Traditional disease-free survival (DFS) does not reflect changes in prognosis over time. Conditional DFS accounts for elapsed time since achieving remission and may provide more relevant prognostic information for patients and clinicians. This study aimed to estimate conditional DFS among patients with ovarian cancer and to evaluate the impact of patient characteristics. Patients and Methods Patients were recruited as part of the Hormones and Ovarian Cancer Prediction case-control study and were included in the current study if they had achieved remission after a diagnosis of cancer of the ovary, fallopian tube, or peritoneum (N = 404). Demographic and lifestyle information was collected at enrollment; disease, treatment, and outcome information was abstracted from medical records. DFS was calculated using the Kaplan-Meier method. Conditional DFS estimates were computed using cumulative DFS estimates. Results Median DFS was 2.54 years (range, 0.03-9.96 years) and 3-year DFS was 48.2%. The probability of surviving an additional 3 years without recurrence, conditioned on having already survived 1, 2, 3, 4, and 5 years after remission, was 63.8%, 80.5%, 90.4%, 97.0%, and 97.7%, respectively. Initial differences in 3-year DFS at time of remission between age, stage, histology, and grade groups decreased over time. Conclusion DFS estimates for patients with ovarian cancer improved dramatically over time, in particular among those with poorer initial prognoses. Conditional DFS is a more relevant measure of prognosis for patients with ovarian cancer who have already achieved a period of remission, and time elapsed since remission should be taken into account when making follow-up care decisions. PMID:25403208

Long-term retention as a function of word concreteness under conditions of free recall.

PubMed

Postman, L; Burns, S

1974-07-01

Acquisition and long-term retention of concrete (C) and abstract (A) words were investigated under conditions of multiple-trial free recall. Both unmixed and mixed lists were used in original learning. Retention was tested either 1 rain or 1 week after attainment of the learning criterion. Acquisition was faster and retention was higher for C than for A words. These differences were more pronounced for mixed than for unmixed lists.

Culture of hESC-derived pancreatic progenitors in alginate-based scaffolds.

PubMed

Formo, Kjetil; Cho, Candy H-H; Vallier, Ludovic; Strand, Berit L

2015-12-01

The effect of alginate-based scaffolds with added basement membrane proteins on the in vitro development of hESC-derived pancreatic progenitors was investigated. Cell clusters were encapsulated in scaffolds containing the basement membrane proteins collagen IV, laminin, fibronectin, or extracellular matrix-derived peptides, and maintained in culture for up to 46 days. The cells remained viable throughout the experiment with no signs of central necrosis. Whereas nonencapsulated cells aggregated into larger clusters, some of which showed signs of morphological changes and tissue organization, the alginate matrix stabilized the cluster size and displayed more homogeneous cell morphologies, allowing culture for long periods of time. For all conditions tested, a stable or declining expression of insulin and PDX1 and an increase in glucagon and somatostatin over time indicated a progressive reduction in beta cell-related gene expression. Alginate scaffolds can provide a chemically defined, xeno-free and easily scalable alternative for culture of pancreatic progenitors. Although no increase in insulin and PDX1 gene expression after alginate-immobilized cell culture was seen in this study, further optimization of the matrix physicochemical and biological properties and of the medium composition may still be a relevant strategy to promote the stabilization or maturation of stem cell-derived beta cells. © 2015 Wiley Periodicals, Inc.

Free in-plane vibration of circular arches.

NASA Technical Reports Server (NTRS)

Veletsos, A. S.; Austin, W. J.; Lopes Pereira, C. A.; Wung, S.-J.

1972-01-01

Numerical data are presented for the natural frequencies and modes of vibration of hinged and fixed, uniform, circular arches vibrating in their own plane, and the effects of the various parameters affecting the response are analyzed. It is shown that the vibrational modes may be almost purely flexural, or almost purely extensional, or the extensional and flexural actions may be strongly coupled. The conditions of occurrence of each type of behavior are defined, and simple approximate formulas are derived; using these formulas, the free vibrational characteristics of arches may be estimated to a satisfactory degree of accuracy for most practical applications. The approach used to derive the approximate formulas may also be applied to arches having other boundary conditions, shapes, or distributions of stiffness and mass.

A comparison between lesions found during meat inspection of finishing pigs raised under organic/free-range conditions and conventional, indoor conditions.

PubMed

Alban, Lis; Petersen, Jesper Valentin; Busch, Marie Erika

2015-01-01

It is often argued that pigs raised under less intensive production conditions - such as organic or free-range - have a higher level of animal welfare compared with conventionally raised pigs. To look into this, an analysis of data from a large Danish abattoir slaughtering organic, free-range, and conventionally raised finishing pigs was undertaken. First, the requirements for each of the three types of production systems were investigated. Next, meat inspection data from a period of 1 year were collected. These covered 201,160 organic/free-range pigs and 1,173,213 conventionally raised pigs. The prevalence of each individual type of lesion was calculated, followed by a statistical comparison between the prevalences in organic/free-range and conventional pigs. Because of the large number of data, the P-value for significance was lowered to P = 0.001, and only biological associations reflecting Odds Ratios above 1.2 or below 0.8 were considered to be of significance. The majority of the lesion types were recorded infrequently (<4%). Only chronic pleuritis was a common finding. A total of 13 lesion types were more frequent among organic/free-range pigs than among conventional pigs - among others old fractures, tail lesions and osteomyelitis. Four lesion types were equally frequent in the two groups: chronic pneumonia, chronic pleuritis, fresh fracture, and abscess in head/ear. Four lesion types were recorded less frequently among organic/free-range pigs compared with conventionally raised pigs. These included abscess in leg/toe, hernia and scar/hock lesion. Possible associations between the individual lesion types and the production systems - including the requirements for each system - are discussed. The results emphasize the importance of using direct animal based parameters when evaluating animal welfare in different types of production systems. Moreover, individual solutions to the health problems observed in a herd should be found, e.g. in collaboration with

Free will and mental disorder: Exploring the relationship

PubMed Central

2010-01-01

A link between mental disorder and freedom is clearly present in the introduction of the fourth edition of the Diagnostic and Statistical Manual of Mental Disorders (DSM-IV). It mentions “an important loss of freedom” as one of the possible defining features of mental disorder. Meanwhile, it remains unclear how “an important loss of freedom” should be understood. In order to get a clearer view on the relationship between mental disorder and (a loss of) freedom, in this article, I will explore the link between mental disorder and free will. I examine two domains in which a connection between mental disorder and free will is present: the philosophy of free will and forensic psychiatry. As it turns out, philosophers of free will frequently refer to mental disorders as conditions that compromise free will and reduce moral responsibility. In addition, in forensic psychiatry, the rationale for the assessment of criminal responsibility is often explained by referring to the fact that mental disorders can compromise free will. Yet, in both domains, it remains unclear in what way free will is compromised by mental disorders. Based on the philosophical debate, I discuss three senses of free will and explore their relevance to mental disorders. I conclude that in order to further clarify the relationship between free will and mental disorder, the accounts of people who have actually experienced the impact of a mental disorder should be included in future research. PMID:20931360

Stability of Pigeon Body Weight under Free-Feeding Conditions

ERIC Educational Resources Information Center

Kangas, Brian D.; Branch, Marc N.

2006-01-01

Increases in regulatory oversight of animal research require verification of effects of standard practices. There are no formal guidelines for establishing free-feeding weights in adult pigeons. In the present study, pigeons were obtained from a commercial supplier, weighed upon arrival, and then held in quarantine for 7 days with free access to…

Synthesis of amino ester-embedded benzimidazoles: a one-pot sequential protocol under metal-free neutral conditions.

PubMed

Roy, Priyabrata; Bodhak, Chandan; Pramanik, Animesh

2017-02-01

A one-pot three-component protocol has been developed for the synthesis of amino ester-embedded benzimidazoles under metal-free neutral conditions. Sequentially, the methodology involves coupling of an amino ester with 1-fluoro-2-nitrobenzene, reduction of the coupled nitroarene by sodium dithionite, and cyclization of the corresponding diamine with an aldehyde.

Free and Cued Memory in relation to Biomarker-Defined Abnormalities in Clinically Normal Older Adults and Those at Risk for Alzheimer’s Disease

PubMed Central

Papp, Kathryn V.; Amariglio, Rebecca E.; Mormino, Elizabeth; Hedden, Trey; Dekhytar, Maria; Johnson, Keith A.; Sperling, Reisa A.; Rentz, Dorene M.

2015-01-01

Objectives Furthering our understanding of the relationship between amyloidosis (Aβ), neurodegeneration (ND), and cognition is imperative for early identification and early intervention of Alzheimer’s disease (AD). However, the subtle cognitive decline differentially associated with each biomarker-defined stage of preclinical AD has yet to be fully characterized. Recent work indicates that different components of memory performance (free and cued recall) may be differentially specific to memory decline in prodromal AD. We sought to examine the relationship between free and cued recall paradigms, in addition to global composites of memory, executive functioning, and processing speed in relation to stages of preclinical AD. Methods A total of 260 clinically normal (CN) older adults (CDR=0) from the Harvard Aging Brain study were grouped according to preclinical AD stages including Stage 0 (Aβ−/ND−), Stage 1 (Aβ+/ND−), Stage 2 (Aβ+/ND+), and suspected non-Alzheimer’s associated pathology (SNAP; Aβ−/ND+). General linear models controlling for age, sex, and education were used to assess for stage-based performance differences on cognitive composites of executive functioning, processing speed, and memory in addition to free and cued delayed recall on the Selective Reminding Test (SRT) and Memory Capacity Test (MCT). Results Global memory performance differed between preclinical stages with Stage 2 performing worse compared with Stage 0. When examining free and cued paradigms by memory test, only the MCT (and not the SRT) revealed group differences. More specifically, Stage 1 was associated with decrements in free recall compared with Stage 0 while Stage 2 was associated with decrements in both free and cued recall. There was a trend for the SNAP group to perform worse on free recall compared with Stage 0. Finally, there was no association between preclinical stage and global composites of executive functioning or processing speed. Conclusions Clinically

Glycaemic Profiles of Children With Overweight and Obesity in Free-living Conditions in Association With Cardiometabolic Risk.

PubMed

Rijks, Jesse; Karnebeek, Kylie; van Dijk, Jan-Willem; Dorenbos, Elke; Gerver, Willem-Jan; Stouthart, Pauline; Plat, Jogchum; Vreugdenhil, Anita

2016-08-18

Insulin resistance is common among children with overweight and obesity. However, knowledge about glucose fluctuations in these children is scarce. This study aims to evaluate glycaemic profiles in children with overweight and obesity in free-living conditions, and to examine the association between glycaemic profiles with insulin resistance and cardiovascular risk parameters. One hundred eleven children with overweight and obesity were included. 48-hour sensor glucose concentrations in free-living conditions, fasting plasma and post-glucose load concentrations, serum lipid and lipoprotein concentrations, homeostatic model assessment of insulin resistance (HOMA-IR), and blood pressure were evaluated. Hyperglycaemic glucose excursions (≥7.8 mmol/L) were observed in 25% (n = 28) of the children. The median sensor glucose concentration was 5.0 (2.7-7.3) mmol/L, and correlated with fasting plasma glucose concentrations (rs = 0.190, p = 0.046), serum insulin concentrations (rs = 0.218, p = 0.021), and HOMA-IR (rs = 0.230, p = 0.015). The hyperglycaemic area under the curve (AUC) correlated with waist circumference z-score (rs = 0.455, p = 0.025), triacylglycerol concentrations (rs = 0.425, p = 0.024), and HOMA-IR (rs = 0.616, p < 0.001). In conclusion, hyperglycaemic glucose excursions are frequently observed in children with overweight and obesity in free-living conditions. Children with insulin resistance had higher median sensor glucose concentrations and a larger hyperglycaemic sensor glucose AUC, which are both associated with specific parameters predicting cardiovascular disease risk.

Elimination of remaining undifferentiated induced pluripotent stem cells in the process of human cardiac cell sheet fabrication using a methionine-free culture condition.

PubMed

Matsuura, Katsuhisa; Kodama, Fumiko; Sugiyama, Kasumi; Shimizu, Tatsuya; Hagiwara, Nobuhisa; Okano, Teruo

2015-03-01

Cardiac tissue engineering is a promising method for regenerative medicine. Although we have developed human cardiac cell sheets by integration of cell sheet-based tissue engineering and scalable bioreactor culture, the risk of contamination by induced pluripotent stem (iPS) cells in cardiac cell sheets remains unresolved. In the present study, we established a novel culture method to fabricate human cardiac cell sheets with a decreased risk of iPS cell contamination while maintaining viabilities of iPS cell-derived cells, including cardiomyocytes and fibroblasts, using a methionine-free culture condition. When cultured in the methionine-free condition, human iPS cells did not survive without feeder cells and could not proliferate or form colonies on feeder cells or in coculture with cells for cardiac cell sheet fabrication. When iPS cell-derived cells after the cardiac differentiation were transiently cultured in the methionine-free condition, gene expression of OCT3/4 and NANOG was downregulated significantly compared with that in the standard culture condition. Furthermore, in fabricated cardiac cell sheets, spontaneous and synchronous beating was observed in the whole area while maintaining or upregulating the expression of various cardiac and extracellular matrix genes. These findings suggest that human iPS cells are methionine dependent and a methionine-free culture condition for cardiac cell sheet fabrication might reduce the risk of iPS cell contamination.

Near-Surface Effects of Free Atmosphere Stratification in Free Convection

NASA Astrophysics Data System (ADS)

Mellado, Juan Pedro; van Heerwaarden, Chiel C.; Garcia, Jade Rachele

2016-04-01

The effect of a linear stratification in the free atmosphere on near-surface properties in a free convective boundary layer (CBL) is investigated by means of direct numerical simulation. We consider two regimes: a neutral stratification regime, which represents a CBL that grows into a residual layer, and a strong stratification regime, which represents the equilibrium (quasi-steady) entrainment regime. We find that the mean buoyancy varies as z^{-1/3}, in agreement with classical similarity theory. However, the root-mean-square (r.m.s.) of the buoyancy fluctuation and the r.m.s. of the vertical velocity vary as z^{-0.45} and ln z, respectively, both in disagreement with theory. These scaling laws are independent of the stratification regime, but the depth over which they are valid depends on the stratification. In the strong stratification regime, this depth is about 20 to 25 % of the CBL depth instead of the commonly used 10 %, which we only observe under neutral conditions. In both regimes, the near-surface flow structure can be interpreted as a hierarchy of circulations attached to the surface. Based on this structure, we define a new near-surface layer in free convection, the plume-merging layer, that is conceptually different from the constant-flux layer. The varying depth of the plume-merging layer depending on the stratification accounts for the varying depth of validity of the scaling laws. These findings imply that the buoyancy transfer law needed in mixed-layer and single-column models is well described by the classical similarity theory, independent of the stratification in the free atmosphere, even though other near-surface properties, such as the r.m.s. of the buoyancy fluctuation and the r.m.s. of the vertical velocity, are inconsistent with that theory.

Transcriptional response of Pasteurella multocida to defined iron sources.

PubMed

Paustian, Michael L; May, Barbara J; Cao, Dongwei; Boley, Daniel; Kapur, Vivek

2002-12-01

Pasteurella multocida was grown in iron-free chemically defined medium supplemented with hemoglobin, transferrin, ferritin, and ferric citrate as iron sources. Whole-genome DNA microarrays were used to monitor global gene expression over seven time points after the addition of the defined iron source to the medium. This resulted in a set of data containing over 338,000 gene expression observations. On average, 12% of P. multocida genes were differentially expressed under any single condition. A majority of these genes encoded P. multocida proteins that were involved in either transport and binding or were annotated as hypothetical proteins. Several trends are evident when the data from different iron sources are compared. In general, only two genes (ptsN and sapD) were expressed at elevated levels under all of the conditions tested. The results also show that genes with increased expression in the presence of hemoglobin did not respond to transferrin or ferritin as an iron source. Correspondingly, genes with increased expression in the transferrin and ferritin experiments were expressed at reduced levels when hemoglobin was supplied as the sole iron source. Finally, the data show that genes that were most responsive to the presence of ferric citrate did not follow a trend similar to that of the other iron sources, suggesting that different pathways respond to inorganic or organic sources of iron in P. multocida. Taken together, our results demonstrate that unique subsets of P. multocida genes are expressed in response to different iron sources and that many of these genes have yet to be functionally characterized.

Imposing the free-slip condition with a continuous forcing immersed boundary method

NASA Astrophysics Data System (ADS)

Kempe, Tobias; Lennartz, Matthias; Schwarz, Stephan; Fröhlich, Jochen

2015-02-01

The numerical simulation of spherical and ellipsoidal bubbles in purified fluids requires the imposition of the free-slip boundary condition at the bubble surface. This paper describes a numerical method for the implementation of free-slip boundary conditions in the context of immersed boundary methods. In contrast to other numerical approaches for multiphase flows, the realization is not straightforward. The reason is that the immersed boundary method treats the liquid as well as the gas phase as a field of constant density and viscosity with a fictitious fluid inside the bubble. The motion of the disperse phase is computed explicitly by solving the momentum balance for each of its elements and is coupled to the continuous phase via additional source terms in the Navier-Stokes equations. The paper starts with illustrating that an ad hoc method is unsuccessful. On this basis, a new method is proposed employing appropriate direct forcing at the bubble surface. A central finding is that with common ratios between the step size of the grid and the bubble diameter, curvature terms need to be accounted for to obtain satisfactory results. The new method is first developed for spherical objects and then extended to generally curved interfaces. This is done by introducing a local coordinate system which approximates the surface in the vicinity of a Lagrangian marker with the help of the two principal curvatures of the surface at this point. The numerical scheme is then validated for spherical and ellipsoidal objects with or without prescribed constant angular velocity. It is shown that the proposed method achieves similar convergence behavior as the method for no-slip boundaries. The results are compared to analytical solutions for creeping flow around a sphere and to numerical reference data obtained on a body-fitted grid. The numerical tests confirm the excellent performance of the proposed method.

9 CFR 71.15 - Movement from quarantined to free area and shipment therefrom; conditions under which permitted.

Code of Federal Regulations, 2014 CFR

2014-01-01

... 9 Animals and Animal Products 1 2014-01-01 2014-01-01 false Movement from quarantined to free area and shipment therefrom; conditions under which permitted. 71.15 Section 71.15 Animals and Animal Products ANIMAL AND PLANT HEALTH INSPECTION SERVICE, DEPARTMENT OF AGRICULTURE INTERSTATE TRANSPORTATION OF...

9 CFR 71.15 - Movement from quarantined to free area and shipment therefrom; conditions under which permitted.

Code of Federal Regulations, 2010 CFR

2010-01-01

... 9 Animals and Animal Products 1 2010-01-01 2010-01-01 false Movement from quarantined to free area and shipment therefrom; conditions under which permitted. 71.15 Section 71.15 Animals and Animal Products ANIMAL AND PLANT HEALTH INSPECTION SERVICE, DEPARTMENT OF AGRICULTURE INTERSTATE TRANSPORTATION OF...

9 CFR 71.15 - Movement from quarantined to free area and shipment therefrom; conditions under which permitted.

Code of Federal Regulations, 2013 CFR

2013-01-01

... 9 Animals and Animal Products 1 2013-01-01 2013-01-01 false Movement from quarantined to free area and shipment therefrom; conditions under which permitted. 71.15 Section 71.15 Animals and Animal Products ANIMAL AND PLANT HEALTH INSPECTION SERVICE, DEPARTMENT OF AGRICULTURE INTERSTATE TRANSPORTATION OF...

9 CFR 71.15 - Movement from quarantined to free area and shipment therefrom; conditions under which permitted.

Code of Federal Regulations, 2011 CFR

2011-01-01

... 9 Animals and Animal Products 1 2011-01-01 2011-01-01 false Movement from quarantined to free area and shipment therefrom; conditions under which permitted. 71.15 Section 71.15 Animals and Animal Products ANIMAL AND PLANT HEALTH INSPECTION SERVICE, DEPARTMENT OF AGRICULTURE INTERSTATE TRANSPORTATION OF...

9 CFR 71.15 - Movement from quarantined to free area and shipment therefrom; conditions under which permitted.

Code of Federal Regulations, 2012 CFR

2012-01-01

... 9 Animals and Animal Products 1 2012-01-01 2012-01-01 false Movement from quarantined to free area and shipment therefrom; conditions under which permitted. 71.15 Section 71.15 Animals and Animal Products ANIMAL AND PLANT HEALTH INSPECTION SERVICE, DEPARTMENT OF AGRICULTURE INTERSTATE TRANSPORTATION OF...

Accuracy of the Yamax CW-701 Pedometer for measuring steps in controlled and free-living conditions

PubMed Central

Coffman, Maren J; Reeve, Charlie L; Butler, Shannon; Keeling, Maiya; Talbot, Laura A

2016-01-01

Objective The Yamax Digi-Walker CW-701 (Yamax CW-701) is a low-cost pedometer that includes a 7-day memory, a 2-week cumulative memory, and automatically resets to zero at midnight. To date, the accuracy of the Yamax CW-701 has not been determined. The purpose of this study was to assess the accuracy of steps recorded by the Yamax CW-701 pedometer compared with actual steps and two other devices. Methods The study was conducted in a campus-based lab and in free-living settings with 22 students, faculty, and staff at a mid-sized university in the Southeastern US. While wearing a Yamax CW-701, Yamax Digi-Walker SW-200, and an ActiGraph GTX3 accelerometer, participants engaged in activities at variable speeds and conditions. To assess accuracy of each device, steps recorded were compared with actual step counts. Statistical tests included paired sample t-tests, percent accuracy, intraclass correlation coefficient, and Bland–Altman plots. Results The Yamax CW-701 demonstrated reliability and concurrent validity during walking at a fast pace and walking on a track, and in free-living conditions. Decreased accuracy was noted walking at a slow pace. Conclusions These findings are consistent with prior research. With most pedometers and accelerometers, adequate force and intensity must be present for a step to register. The Yamax CW-701 is accurate in recording steps taken while walking at a fast pace and in free-living settings. PMID:29942555

Accuracy of the Yamax CW-701 Pedometer for measuring steps in controlled and free-living conditions.

PubMed

Coffman, Maren J; Reeve, Charlie L; Butler, Shannon; Keeling, Maiya; Talbot, Laura A

2016-01-01

The Yamax Digi-Walker CW-701 (Yamax CW-701) is a low-cost pedometer that includes a 7-day memory, a 2-week cumulative memory, and automatically resets to zero at midnight. To date, the accuracy of the Yamax CW-701 has not been determined. The purpose of this study was to assess the accuracy of steps recorded by the Yamax CW-701 pedometer compared with actual steps and two other devices. The study was conducted in a campus-based lab and in free-living settings with 22 students, faculty, and staff at a mid-sized university in the Southeastern US. While wearing a Yamax CW-701, Yamax Digi-Walker SW-200, and an ActiGraph GTX3 accelerometer, participants engaged in activities at variable speeds and conditions. To assess accuracy of each device, steps recorded were compared with actual step counts. Statistical tests included paired sample t -tests, percent accuracy, intraclass correlation coefficient, and Bland-Altman plots. The Yamax CW-701 demonstrated reliability and concurrent validity during walking at a fast pace and walking on a track, and in free-living conditions. Decreased accuracy was noted walking at a slow pace. These findings are consistent with prior research. With most pedometers and accelerometers, adequate force and intensity must be present for a step to register. The Yamax CW-701 is accurate in recording steps taken while walking at a fast pace and in free-living settings.

Energetic Assessment of the Nonexercise Activities under Free-Living Conditions.

PubMed

Sun, Shijie; Tang, Qiang; Quan, Haiying; Lu, Qi; Sun, Ming; Zhang, Kuan

2016-01-01

Nonexercise activities (NAs) are common types of physical activity in daily life and critical component in energy expenditure. However, energetic assessment of NA, particularly in free-living subjects, is a technical challenge. In this study, mechanical modeling and portable device were used to evaluate five common types of NA in daily life: sit to stand, lie to sit, bowing while standing, squat, and right leg over left. A human indirect calorimeter was used to measure the activity energy expenditure of NA. Mechanical work and mechanical efficiency of NA were calculated for mechanical modeling. Thirty-two male subjects were recruited for the study (20 subjects for the development of models and 12 subjects for evaluation of models). The average (mean ± SD) mechanical work of 5 NAs was 2.31 ± 0.50, 2.88 ± 0.57, 1.75 ± 0.55, 3.96 ± 1.25, and 1.25 ± 0.51 J/kg·m, respectively. The mean mechanical efficiencies of those activities were 22.0 ± 3.3%, 26.5 ± 5.1%, 19.8 ± 3.7%, 24.0 ± 5.5%, and 26.3 ± 5.5%. The activity energy expenditure estimated by the models was not significantly different from the measurements by the calorimeter (p > 0.05) with accuracies of 102.2 ± 20.7%, 103.7 ± 25.8%, 105.6 ± 14.6%, 101.1 ± 28.0%, and 95.8 ± 20.7%, respectively, for those activities. These findings suggest that the mechanical models combined with a portable device can provide an alternative method for the energetic analysis of nonexercise activities under free-living condition.

Prevalence of gastrointestinal helminth infections in free-range laying hens under mountain farming production conditions.

PubMed

Wuthijaree, K; Lambertz, C; Gauly, M

2017-12-01

1. A cross-sectional study was conducted from September 2015 to July 2016 in South Tyrol, Northern Italy to examine the prevalence of gastrointestinal helminths in free-range laying hens under mountain farming production conditions. 2. A total of 280 laying hens from 14 free-range mountain farms (4 organic, 10 conventional) were randomly collected at the end of the laying period. Faecal samples were taken to analyse faecal egg counts (FEC) and faecal oocyst counts (FOC). The gastrointestinal tracts were removed post mortem and examined for the presence of helminths. 3. In faeces, FEC values averaged 258 eggs per g of faeces, which were dominated by Ascaridia galli and Heterakis gallinarum. Mean FOC was 80 oocysts/g. In the gastrointestinal tract, at least one nematode species was found in 99.3% of the examined hens. H. gallinarum was the most prevalent nematode (95.7%), followed by Capillaria spp. (66.8%) and A. galli (63.6%). Thirty per cent of the chickens were infected with cestodes (tapeworms). Correlation coefficients between worm counts of H. gallinarum, Capillaria spp. and A. galli ranged from 0.41 to 0.51. 5. The helminth prevalence did not differ between conventional and organic farms, whereas total worm burden was higher in organic compared with conventional farms (318.9 vs. 112.0). Prevalence and infection intensity did not differ between farms that used anthelmintic treatments and those that did not. 6. In conclusion, free-range laying hens under the studied mountain farming conditions are at high risk of nematode infection, especially in organic systems. The vast majority of hens are subclinical infected with at least one helminth species.

Solutions of the Helmholtz equation with boundary conditions for force-free magnetic fields

NASA Technical Reports Server (NTRS)

Rasband, S. N.; Turner, L.

1981-01-01

It is shown that the solution, with one ignorable coordinate, for the Taylor minimum energy state (resulting in a force-free magnetic field) in either a straight cylindrical or a toroidal geometry with arbitrary cross section can be reduced to the solution of either an inhomogeneous Helmholtz equation or a Grad-Shafranov equation with simple boundary conditions. Standard Green's function theory is, therefore, applicable. Detailed solutions are presented for the Taylor state in toroidal and cylindrical domains having a rectangular cross section. The focus is on solutions corresponding to the continuous eigenvalue spectra. Singular behavior at 90 deg corners is explored in detail.

Defining Putative Glycan Cancer Biomarkers by Mass Spectrometry

PubMed Central

Mechref, Yehia; Hu, Yunli; Garcia, Aldo; Hussein, Ahmed

2013-01-01

Summary For decades, the association between aberrant glycosylation and many types of cancers has been shown. However, defining the changes of glycan structures has not been demonstrated until recently. This has been facilitated by the major advances in mass spectrometry and separation science which allowed the detailed characterization of glycan changes associated with cancer. Mass spectrometry glycomics methods have been successfully employed to compare the glycomic profiles of different human specimen collected from disease-free individuals and patients with cancer. Additionally, comparing the glycomic profiles of glycoproteins purified from specimen collected from disease-free individuals and patients with cancer has also been performed. These types of glycan analyses employing mass spectrometry or liquid-chromatography mass spectrometry allowed the characterization of native, labeled, and permethylated glycans. This review discusses the different glycomic and glycoproteomic methods employed for defining glycans as cancer biomarkers of different organs, including breast, colon, esophagus, liver, lung, ovarian, pancreas and prostate. PMID:23157355

Efficient generation of transgene- and feeder-free induced pluripotent stem cells from human dental mesenchymal stem cells and their chemically defined differentiation into cardiomyocytes.

PubMed

Tan, Xiaobing; Dai, Qingli; Guo, Tao; Xu, Jingshu; Dai, Qingyuan

2018-01-22

Advance in stem cell research resulted in several processes to generate induced pluripotent stem cells (iPSCs) from adult somatic cells. In our previous study, the reprogramming of iPSCs from human dental mesenchymal stem cells (MSCs) including SCAP and DPSCs, has been reported. Herein, safe iPSCs were reprogrammed from SCAP and DPSCs using non-integrating RNA virus vector, which is an RNA virus carrying no risk of altering host genome. DPSCs- and SCAP-derived iPSCs exhibited the characteristics of the classical morphology with human embryonic stem cells (hESCs) without integration of foreign genes, indicating the potential of their clinical application. Moreover, induced PSCs showed the capacity of self-renewal and differentiation into cardiac myocytes. We have achieved the differentiation of hiPSCs to cardiomyocytes lineage under serum and feeder-free conditions, using a chemically defined medium CDM3. In CDM3, hiPSCs differentiation is highly generating cardiomyocytes. The results showed this protocol produced contractile sheets of up to 97.2% TNNT2 cardiomyocytes after purification. Furthermore, derived hiPSCs differentiated to mature cells of the three embryonic germ layers in vivo and in vitro of beating cardiomyocytes. The above whole protocol enables the generation of large scale of highly pure cardiomyocytes as needed for cellular therapy. Copyright © 2017. Published by Elsevier Inc.

Evaluation of selected properties of gluten-free instant gruels processed under various extrusion-cook- ing conditions.

PubMed

Kręcisz, Magdalena; Wójtowicz, Agnieszka

2017-01-01

For consumers suffering with gluten intolerance, the only way to manage the condition is to avoid foods which are high in gluten. Instant gruels, processed from gluten-free corn and rice by extrusion cooking, could be used as a ready meal both for children and for adults on a gluten-free diet. The aim of the study was to evaluate the effects of various processing conditions on selected characteristics of corn-rice instant gruels. Corn-rice mixtures (75:25 and 50:50) were processed at 12, 14, 16 and 18% of initial moisture content, using an extruder with screw speeds of 80, 100 and 120 rpm. Bulk density, water absorption and solubility, gel formation, color and sensory characteristics were assessed, under various pro- cessing conditions and with various corn:rice ratios. The composition of the raw materials, initial moisture content and screw speed applied during processing affected the characteristics of the corn-rice extruded instant gruels. Increasing the amount of rice in the recipe from 25 to 50% resulted in decreased bulk density, water solubility, volumetric gel formation ability and b* value. Increasing the initial moisture content increased the bulk density, L*, a* and b* intensity, and gel formation index values of extrudates made with a 75:25 corn-rice recipe. Increased rpm increased extrudate solubility and water absorption, if the initial moisture content was higher than 14%. The highest scores for overall acceptability were found for milk suspensions of 75:25 and 50:50 corn-rice instant gruels processed at 12 and 14% of initial moisture content, at 120 rpm. Corn-rice instant gruels can be successfully produced by extrusion-cooking. Variable param- eters, like the initial moisture content of raw materials or screw speed during processing significantly affected the properties of the products. An understanding of the effects of processing conditions on some qualities of extruded instant gruels allows more desirable products to be created. Moreover, the

Defining the free-energy landscape of curvature-inducing proteins on membrane bilayers

PubMed Central

Tourdot, Richard W.; Ramakrishnan, N.; Radhakrishnan, Ravi

2015-01-01

Curvature-sensing and curvature-remodeling proteins, such as Amphiphysin, Epsin, and Exo70, are known to reshape cell membranes, and this remodeling event is essential for key biophysical processes such as tubulation, exocytosis, and endocytosis. Curvature-inducing proteins can act as curvature sensors; they aggregate to membrane regions matching their intrinsic curvature; as well as induce curvature in cell membranes to stabilize emergent high curvature, nonspherical, structures such as tubules, discs, and caveolae. A definitive understanding of the interplay between protein recruitment and migration, the evolution of membrane curvature, and membrane morphological transitions is emerging but remains incomplete. Here, within a continuum framework and using the machinery of Monte Carlo simulations, we introduce and compare three free-energy methods to delineate the free-energy landscape of curvature-inducing proteins on bilayer membranes. We demonstrate the utility of the Widom test particle (or field) insertion methodology in computing the excess chemical potentials associated with curvature-inducing proteins on the membrane—in particular, we use this method to track the onset of morphological transitions in the membrane at elevated protein densities. We validate this approach by comparing the results from the Widom method with those of thermodynamic integration and Bennett acceptance ratio methods. Furthermore, the predictions from the Widom method have been tested against analytical calculations of the excess chemical potential at infinite dilution. Our results are useful in precisely quantifying the free-energy landscape, and also in determining the phase boundaries associated with curvature-induction, curvature-sensing, and morphological transitions. This approach can be extended to studies exploring the role of thermal fluctuations and other external (control) variables, such as membrane excess area, in shaping curvature-mediated interactions on bilayer

Defining the free-energy landscape of curvature-inducing proteins on membrane bilayers.

PubMed

Tourdot, Richard W; Ramakrishnan, N; Radhakrishnan, Ravi

2014-08-01

Curvature-sensing and curvature-remodeling proteins, such as Amphiphysin, Epsin, and Exo70, are known to reshape cell membranes, and this remodeling event is essential for key biophysical processes such as tubulation, exocytosis, and endocytosis. Curvature-inducing proteins can act as curvature sensors; they aggregate to membrane regions matching their intrinsic curvature; as well as induce curvature in cell membranes to stabilize emergent high curvature, nonspherical, structures such as tubules, discs, and caveolae. A definitive understanding of the interplay between protein recruitment and migration, the evolution of membrane curvature, and membrane morphological transitions is emerging but remains incomplete. Here, within a continuum framework and using the machinery of Monte Carlo simulations, we introduce and compare three free-energy methods to delineate the free-energy landscape of curvature-inducing proteins on bilayer membranes. We demonstrate the utility of the Widom test particle (or field) insertion methodology in computing the excess chemical potentials associated with curvature-inducing proteins on the membrane-in particular, we use this method to track the onset of morphological transitions in the membrane at elevated protein densities. We validate this approach by comparing the results from the Widom method with those of thermodynamic integration and Bennett acceptance ratio methods. Furthermore, the predictions from the Widom method have been tested against analytical calculations of the excess chemical potential at infinite dilution. Our results are useful in precisely quantifying the free-energy landscape, and also in determining the phase boundaries associated with curvature-induction, curvature-sensing, and morphological transitions. This approach can be extended to studies exploring the role of thermal fluctuations and other external (control) variables, such as membrane excess area, in shaping curvature-mediated interactions on bilayer

Time-of-flight measurement of ionic species generated during ablation for optimization of focusing condition at free-electron laser beamline

NASA Astrophysics Data System (ADS)

Senba, Y.; Nagasono, M.; Koyama, T.; Yumoto, H.; Ohashi, H.; Tono, K.; Togashi, T.; Inubushi, Y.; Sato, T.; Yabashi, M.; Ishikawa, T.

2013-03-01

Optimization of focusing conditions is important in free-electron laser applications. A time-of-flight mass analyzer has been designed and constructed for this purpose. The time-of-flight spectra of ionic species evolved from laser ablation of gold were measured. The yields of ionic species showed strong correlations with free-electron-laser intensity. This method conveniently allows for direct estimation of laser intensity on sample and determination of focusing position.

Reducing Spread in Climate Model Projections of a September Ice-Free Arctic

NASA Technical Reports Server (NTRS)

Liu, Jiping; Song, Mirong; Horton, Radley M.; Hu, Yongyun

2013-01-01

This paper addresses the specter of a September ice-free Arctic in the 21st century using newly available simulations from the Coupled Model Intercomparison Project Phase 5 (CMIP5). We find that large spread in the projected timing of the September ice-free Arctic in 30 CMIP5 models is associated at least as much with different atmospheric model components as with initial conditions. Here we reduce the spread in the timing of an ice-free state using two different approaches for the 30 CMIP5 models: (i) model selection based on the ability to reproduce the observed sea ice climatology and variability since 1979 and (ii) constrained estimation based on the strong and persistent relationship between present and future sea ice conditions. Results from the two approaches show good agreement. Under a high-emission scenario both approaches project that September ice extent will drop to approx. 1.7 million sq km in the mid 2040s and reach the ice-free state (defined as 1 million sq km) in 2054-2058. Under a medium-mitigation scenario, both approaches project a decrease to approx.1.7 million sq km in the early 2060s, followed by a leveling off in the ice extent.

Clinical perspective of cell-free DNA testing for fetal aneuploidies.

PubMed

Gratacós, Eduard; Nicolaides, Kypros

2014-01-01

Cell-free DNA testing in maternal blood provides the most effective method of screening for trisomy 21, with a reported detection rate of 99% and a false positive rate of less than 0.1%. After many years of research, this method is now commercially available and is carried out in an increasing number of patients, and there is an expanding number of conditions that can be screened for. However, the application of these methods in clinical practice requires a careful analysis. Current first-trimester screening strategies are based on a complex combination of tests, aiming at detecting fetal defects and predicting the risk of main pregnancy complications. It is therefore necessary to define the optimal way of combining cell-free DNA testing with current first-trimester screening methods. In this concise review we describe the basis of cell-free DNA testing and discuss the potential approaches for its implementation in combination with current tests in the first trimester. © 2014 S. Karger AG, Basel.

Effects of intentionality and subliminal information in free-choices to inhibit.

PubMed

Dall'Acqua, Tommaso; Begliomini, Chiara; Motta, Raffaella; Miotto, Diego; Castiello, Umberto

2018-01-31

Stopping an action at the very last moment is an important feature of human behavioural flexibility. Intentional inhibition has been defined as the ability to inhibit an action on the basis of an internal decision process. Without this ability, actions would be impulsive and would leave little space to correct misguided decisions. Previous research suggests that making a choice between action alternatives activates a specific "choice network" that includes the rostral cingulate zone (RCZ), the anterior insula (AI), the dorsolateral prefrontal cortex (DLPFC) and the inferior parietal lobe (IPL). The activity of this network has shown to be influenced by non-conscious (subliminal) stimuli. In this study, we tested whether the same regions are recruited by free-choices to inhibit and modulated by unconscious information as reported in the case of free-choices to act. Using functional magnetic resonance imaging (fMRI) we manipulated the degree of 'freedom' of the choice between acting and inhibiting an action by introducing explicit cues or leaving the participants free to choose between action alternatives. We included subliminal masked primes to test whether responses to targets were facilitated and/or obstructed by conditions of congruency and incongruency between primes and targets. Our findings confirmed higher activation of the "choice network" in free-choice trials when compared to cued choices. However subliminal priming failed to significantly influence participants' responses, in free-choice conditions. Copyright © 2017 Elsevier Ltd. All rights reserved.

Adaptation of the human Cell Line Activation Test (h-CLAT) to Animal-Product-Free Conditions.

PubMed

Edwards, Alexander; Roscoe, Lottie; Longmore, Christopher; Bailey, Fiona; Sim, Bushra; Treasure, Carol

2018-06-13

Skin sensitisers are substances that can elicit allergic responses following skin contact and the process by which this occurs is described as skin sensitisation. Skin sensitisation is defined as a series of key events, that form an adverse outcome pathway (AOP). Key event three in the AOP is dendritic cell activation that can be modelled by the human Cell Line Activation Test (h-CLAT) and is typified by changes in cell surface markers CD54 and CD86 in dendritic cells. The h-CLAT is accepted at a regulatory level (OECD Test-Guideline (TG)442E) and can be used to assess skin sensitisation potential as part of an integrated approach to testing and assessment (IATA). Stakeholders in the cosmetics and chemical industries have scientific and ethical concerns relating to use of animal derived material and have communicated a strong preference for fully human based in vitro methods. Therefore, we adapted the h-CLAT to animal-product-free conditions and validated the adapted method with the proficiency panel substances in Annex II of TG442E, using 3 independent batches of pooled human serum. The modified method showed equivalence to the validated reference method (VRM), as all proficiency substances were correctly classified. Comparable values for CV75 (concentration yielding 75% cell viability), EC150 and EC200 (concentration yielding RFI of ≥150 for CD86 and ≥200 for CD54) were obtained. Data generated using the adapted method may be used in European REACH submissions, provided the proficiency data is included. We are seeking formal inclusion of the adaptation into TG442E, enabling compliance with global regulations.

Effects of shelter type, early environmental enrichment and weather conditions on free-range behaviour of slow-growing broiler chickens.

PubMed

Stadig, L M; Rodenburg, T B; Ampe, B; Reubens, B; Tuyttens, F A M

2017-06-01

Free-range use by broiler chickens is often limited, whereas better use of the free-range area could benefit animal welfare. Use of free-range areas could be stimulated by more appropriate shelter or environmental enrichment (by decreasing birds' fearfulness). This study aimed to assess the effects of shelter type, early environmental enrichment and weather conditions on free-range use. Three production rounds with 440 slow-growing broiler chickens (Sasso T451) were carried out. Birds were housed indoors in four groups (two with males, two with females) from days 0 to 25, during which two of the groups received environmental enrichment. At day 23 birds' fearfulness was assessed with a tonic immobility (TI) test (n=100). At day 25 all birds were moved (in mixed-sex groups) to mobile houses, and provided with free-range access from day 28 onwards. Each group could access a range consisting for 50% of grassland with 21 artificial shelters (ASs, wooden A-frames) and for 50% of short rotation coppice (SRC) with willow (dense vegetation). Free-range use was recorded by live observations at 0900, 1300 and 1700 h for 15 to 21 days between days 28 and 63. For each bird observed outside the shelter type (AS or SRC), distance from the house (0 to 2, 2 to 5, >5 m) and its behaviour (only rounds 2 and 3) were recorded. Weather conditions were recorded by four weather stations. On average, 27.1% of the birds were observed outside at any given moment of observation. Early environmental enrichment did not decrease fearfulness as measured by the TI test. It only had a minor effect on the percentage of birds outside (0.4% more birds outside). At all distances from the house, SRC was preferred over AS. In AS, areas closer to the house were preferred over farther ones, in SRC this was less pronounced. Free-range use increased with age and temperature and decreased with wind speed. In AS, rainfall and decreasing solar radiation were related to finding more birds outside, whereas the

MICROWAVE-FACILITATED MULTICOMPONENT REACTIONS UNDER SOLVENT-FREE CONDITIONS

EPA Science Inventory

The application of microwave-expedited solvent-free synthetic protocols in multi-component (MCC) reactions will be exemplified by several condensation and cyclization reactions including the rapid one-pot assembly of heterocyclic compounds from in situ generated intermediates. R...

ROCK inhibitor is not required for embryoid body formation from singularized human embryonic stem cells.

PubMed

Pettinato, Giuseppe; Vanden Berg-Foels, Wendy S; Zhang, Ning; Wen, Xuejun

2014-01-01

We report a technology to form human embryoid bodies (hEBs) from singularized human embryonic stem cells (hESCs) without the use of the p160 rho-associated coiled-coil kinase inhibitor (ROCKi) or centrifugation (spin). hEB formation was tested under four conditions: +ROCKi/+spin, +ROCKi/-spin, -ROCKi/+spin, and -ROCKi/-spin. Cell suspensions of BG01V/hOG and H9 hESC lines were pipetted into non-adherent hydrogel substrates containing defined microwell arrays. hEBs of consistent size and spherical geometry can be formed in each of the four conditions, including the -ROCKi/-spin condition. The hEBs formed under the -ROCKi/-spin condition differentiated to develop the three embryonic germ layers and tissues derived from each of the germ layers. This simplified hEB production technique offers homogeneity in hEB size and shape to support synchronous differentiation, elimination of the ROCKi xeno-factor and rate-limiting centrifugation treatment, and low-cost scalability, which will directly support automated, large-scale production of hEBs and hESC-derived cells needed for clinical, research, or therapeutic applications.

Eclipse-Free-Time Assessment Tool for IRIS

NASA Technical Reports Server (NTRS)

Eagle, David

2012-01-01

IRIS_EFT is a scientific simulation that can be used to perform an Eclipse-Free- Time (EFT) assessment of IRIS (Infrared Imaging Surveyor) mission orbits. EFT is defined to be those time intervals longer than one day during which the IRIS spacecraft is not in the Earth s shadow. Program IRIS_EFT implements a special perturbation of orbital motion to numerically integrate Cowell's form of the system of differential equations. Shadow conditions are predicted by embedding this integrator within Brent s method for finding the root of a nonlinear equation. The IRIS_EFT software models the effects of the following types of orbit perturbations on the long-term evolution and shadow characteristics of IRIS mission orbits. (1) Non-spherical Earth gravity, (2) Atmospheric drag, (3) Point-mass gravity of the Sun, and (4) Point-mass gravity of the Moon. The objective of this effort was to create an in-house computer program that would perform eclipse-free-time analysis. of candidate IRIS spacecraft mission orbits in an accurate and timely fashion. The software is a suite of Fortran subroutines and data files organized as a "computational" engine that is used to accurately predict the long-term orbit evolution of IRIS mission orbits while searching for Earth shadow conditions.

Six-year clinical performance of etch-and-rinse and self-etch adhesives.

PubMed

Boushell, Lee W; Heymann, Harald O; Ritter, Andre V; Sturdevant, John R; Swift, Edward J; Wilder, Aldridge D; Chung, Yunro; Lambert, Cynthia A; Walter, Ricardo

2016-09-01

To evaluate the 6-year clinical performance of Xeno IV, Xeno III, and XP Bond adhesives in the restoration of non-carious cervical lesions (NCCLs). This was a randomized controlled clinical trial where 39 participants met the inclusion/exclusion criteria and were enrolled. Lesions restored were notch-shaped NCCLs. Prior to adhesive procedures, NCCLs were roughened. No enamel bevel was placed and no mechanical retention was created. Adhesive systems were applied following manufacturer's instructions and the NCCLs were restored with composite resin (TPH(3)). Restorations were finished immediately after placement and scored with regard to retention, marginal discoloration, marginal adaptation, sensitivity, and secondary caries using modified USPHS criteria. Descriptive statistics were performed. Logistic regression models were performed for each outcome separately with compound symmetry correlation structure where teeth were clustered by participants. All analyses were conducted using SAS 9.2. The 6-year recall rate was 77.5% of the restorations (76.9% of the participants). No statistical differences were found between adhesives for retention, marginal discoloration, and marginal adaptation. Restorations failed by loss of retention (16.7%, 27.6%, and 11.8% of Xeno IV, Xeno III, and XP Bond restorations, respectively) and marginal discoloration (7.4% of Xeno IV restorations). For every unit increase in restoration volume it was 1.31 (95%C.I. 1.05, 1.63, P=0.01) times more likely that the restoration retention would be maintained. The tested adhesive systems presented similar clinical performance after six years of service, with annual failure rates of 2.8%, 4.6%, and 2.0% for Xeno IV, Xeno III, and XP Bond, respectively. Copyright © 2016 The Academy of Dental Materials. Published by Elsevier Ltd. All rights reserved.

Efficacy of manual versus free-weight training to improve maximal strength and performance for microgravity conditions.

PubMed

Behringer, Michael; Schüren, Thomas; McCourt, Molly; Mester, Joachim

2016-01-01

We tested a simple and compact device designed for manual resistance training in conditions of microgravity (Self-Powered Rope Trainer Duo (SPoRT Duo)) to increase muscle performance. Twenty-four participants (20.8 ± 2.1 years) were randomly assigned to a manual resistance group (n = 12) and a free-weight group (n = 12). Participants performed eight exercises (three sets; 8-12 efforts) either with free weights or the SPoRT Duo twice a week for 6 weeks. Maximal isometric force of trunk flexion, back extension and chest press increased (P at least 0.01, d at least 0.52) both in the manual resistance group (18.4% ± 15.0%; 32.7% ± 22.7%; 15.3% ± 9.7%) and free-weight group (18.0% ± 13.9%; 26.6% ± 28.9%; 13.3% ± 7.6%). The change in maximal isometric force of wide grip row in both groups (d at best 0.38) did not reach statistical significance (P at best 0.08). The squat one-repetition-maximum increased in the manual resistance group (29.8% ± 22.1%) and the free-weight group (32.4% ± 26.6%). Jump height, determined by a jump-and-reach test, increased in the free-weight group (9.8% ± 13.2%) but not in the manual resistance group (2.0% ± 8.5%). Manual resistance training was equally effective in increasing strength as traditional resistance training with free weights. This apparatus is a useful addition to current in-flight exercise systems.

The Formation of Carbide-Free Bainite in High-Carbon High-Silicon Steel under Isothermal Conditions

NASA Astrophysics Data System (ADS)

Tereshchenko, N. A.; Yakovleva, I. L.; Mirzaev, D. A.; Buldashev, I. V.

2017-12-01

It is shown that a carbide-free bainite structure can be formed in high-carbon steel of the Fe-Si-Mn-Cr-V system using a traditional furnace facility. The structural aspects of bainitic transformation developing under isothermal conditions at 300°C have been studied by the methods of X-ray diffraction and transmission electron microscopy. Orientation relationships between crystalline lattices of γ and α phases have been established. A superequilibrium carbon concentration in the bainite α phase has been determined.

78 FR 47154 - Food Labeling; Gluten-Free Labeling of Foods

Federal Register 2010, 2011, 2012, 2013, 2014

2013-08-05

...The Food and Drug Administration (FDA or we) is issuing a final rule to define the term ``gluten-free'' for voluntary use in the labeling of foods. The final rule defines the term ``gluten-free'' to mean that the food bearing the claim does not contain an ingredient that is a gluten-containing grain (e.g., spelt wheat); an ingredient that is derived from a gluten-containing grain and that has not been processed to remove gluten (e.g., wheat flour); or an ingredient that is derived from a gluten-containing grain and that has been processed to remove gluten (e.g., wheat starch), if the use of that ingredient results in the presence of 20 parts per million (ppm) or more gluten in the food (i.e., 20 milligrams (mg) or more gluten per kilogram (kg) of food); or inherently does not contain gluten; and that any unavoidable presence of gluten in the food is below 20 ppm gluten (i.e., below 20 mg gluten per kg of food). A food that bears the claim ``no gluten,'' ``free of gluten,'' or ``without gluten'' in its labeling and fails to meet the requirements for a ``gluten-free'' claim will be deemed to be misbranded. In addition, a food whose labeling includes the term ``wheat'' in the ingredient list or in a separate ``Contains wheat'' statement as required by a section of the Federal Food, Drug, and Cosmetic Act (the FD&C Act) and also bears the claim ``gluten-free'' will be deemed to be misbranded unless its labeling also bears additional language clarifying that the wheat has been processed to allow the food to meet FDA requirements for a ``gluten-free'' claim. Establishing a definition of the term ``gluten-free'' and uniform conditions for its use in food labeling will help ensure that individuals with celiac disease are not misled and are provided with truthful and accurate information with respect to foods so labeled. We are issuing the final rule under the Food Allergen Labeling and Consumer Protection Act of 2004 (FALCPA).

Walk this way: validity evidence of iphone health application step count in laboratory and free-living conditions.

PubMed

Duncan, Markus J; Wunderlich, Kelly; Zhao, Yingying; Faulkner, Guy

2018-08-01

Several attempts have been made to demonstrate the accuracy of the iPhone pedometer function in laboratory test conditions. However, no studies have attempted to evaluate evidence of convergent validity of the iPhone step counts as a surveillance tool in the field. This study takes a pragmatic approach to evaluating Health application derived iPhone step counts by measuring accuracy of a standardized criterion iPhone SE and a heterogeneous sample of participant owned iPhones (6 or newer) in a laboratory condition, as well as comparing personal iPhones to accelerometer derived steps in a free-living test. During lab tests, criterion and personal iPhones differed from manually counted steps by a mean bias of less than ±5% when walking at 5km/h, 7.5km/h and 10km/h on a treadmill, which is generally considered acceptable for pedometers. In the free-living condition steps differed by a mean bias of 21.5% or 1340 steps/day when averaged across observation days. Researchers should be cautioned in considering the use of iPhone models as a research grade pedometer for physical activity surveillance or evaluation, likely due to the iPhone not being continually carried by participants; if compliance can be maximized then the iPhone might be suitable.

Optimizing blood collection, transport and storage conditions for cell free DNA increases access to prenatal testing.

PubMed

Wong, David; Moturi, Sharmili; Angkachatchai, Vach; Mueller, Reinhold; DeSantis, Grace; van den Boom, Dirk; Ehrich, Mathias

2013-08-01

Fetal mutations and fetal chromosomal abnormalities can be detected by molecular analysis of circulating cell free fetal DNA (ccffDNA) from maternal plasma. This comprehensive study was aimed to investigate and verify blood collection and blood shipping conditions that enable Noninvasive Prenatal Testing. Specifically, the impact of shipping and storage on the stability and concentration of circulating cell-free DNA (ccfDNA) in Streck® Cell-Free DNA™ Blood Collection Tubes (Streck BCTs, Streck, Omaha NE). These BCTs were designed to minimize cellular degradation, and thus effectively prevent dilution of fetal ccf DNA by maternal genomic DNA, was evaluated. Peripheral venous maternal blood was collected into Streck BCTs to investigate four aspects of handling and processing conditions: (1) time from blood draw to plasma processing; (2) storage temperature; (3) mechanical stress; and (4) lot-to-lot tube variations. Maternal blood stored in Streck BCTs for up to 7 days at ambient temperature provides stable concentrations of ccffDNA. The amount of fetal DNA did not change over a broad range of storage temperatures (4°C, 23°C, 37°C, 40°C), but the amount of total (largely maternal) DNA increased in samples stored at 23°C and above, indicating maternal cell degradation and genomic DNA release at elevated temperatures. Shipping maternal blood in Streck BCTs, did not affect sample quality. Maternal plasma DNA stabilized for 0 to 7 days in Streck BCTs can be used for non-invasive prenatal molecular applications, when temperatures are maintained within the broad parameters assessed in this study. Copyright © 2013 The Authors. Published by Elsevier Inc. All rights reserved.

A new method for estimating UV fluxes at ground level in cloud-free conditions

NASA Astrophysics Data System (ADS)

Wandji Nyamsi, William; Pitkänen, Mikko R. A.; Aoun, Youva; Blanc, Philippe; Heikkilä, Anu; Lakkala, Kaisa; Bernhard, Germar; Koskela, Tapani; Lindfors, Anders V.; Arola, Antti; Wald, Lucien

2017-12-01

A new method has been developed to estimate the global and direct solar irradiance in the UV-A and UV-B at ground level in cloud-free conditions. It is based on a resampling technique applied to the results of the k-distribution method and the correlated-k approximation of Kato et al. (1999) over the UV band. Its inputs are the aerosol properties and total column ozone that are produced by the Copernicus Atmosphere Monitoring Service (CAMS). The estimates from this new method have been compared to instantaneous measurements of global UV irradiances made in cloud-free conditions at five stations at high latitudes in various climates. For the UV-A irradiance, the bias ranges between -0.8 W m-2 (-3 % of the mean of all data) and -0.2 W m-2 (-1 %). The root mean square error (RMSE) ranges from 1.1 W m-2 (6 %) to 1.9 W m-2 (9 %). The coefficient of determination R2 is greater than 0.98. The bias for UV-B is between -0.04 W m-2 (-4 %) and 0.08 W m-2 (+13 %) and the RMSE is 0.1 W m-2 (between 12 and 18 %). R2 ranges between 0.97 and 0.99. This work demonstrates the quality of the proposed method combined with the CAMS products. Improvements, especially in the modeling of the reflectivity of the Earth's surface in the UV region, are necessary prior to its inclusion into an operational tool.

Free-standing polyelectrolyte membranes made of chitosan and alginate

PubMed Central

Caridade, Sofia G.; Monge, Claire; Gilde, Flora; Boudou, Thomas; Mano, João F.; Picart, Catherine

2014-01-01

Free-standing films have increasing applications in the biomedical field as drug delivery systems, for wound healing and tissue engineering. Here, we prepared free-standing membranes by the layer-by-layer assembly of chitosan and alginate, two widely used biomaterials. Our aim was to produce thick membrane, to study the permeation of model drugs and the adhesion of muscle cells. We first defined the optimal growth conditions in terms of pH and alginate concentration. The membranes could be easily detached from polystyrene or polypropylene substrate without any post-processing step. They dry thickness was varied over a large range from 4 to 35 μm. A two-fold swelling was observed by confocal microscopy when they were immersed in PBS. In addition, we quantified the permeation of model drugs (fluorescent dextrans) through the free standing membrane, which depended on the dextran molecular weight. Finally, we showed that myoblast cells exhibited a preferential adhesion on the alginate-ending membrane as compared to the chitosan-ending membrane or to the substrate side. PMID:23590116

A numerical solution to define channel heads and hillslope parameters from digital topography of glacially conditioned catchments

NASA Astrophysics Data System (ADS)

Salcher, Bernhard; Baumann, Sebastian; Kober, Florian; Robl, Jörg; Heiniger, Lukas

2016-04-01

The analysis of the slope-area relationship in bedrock streams is a common way for discriminating the channel from the hillslope domain and associated landscape processes. Spatial variations of these domains are important indicators of landscape change. In fluvial catchments, this relationship is a function of contributing drainage area, channel slope and the threshold drainage area for fluvial erosion. The resulting pattern is related to climate, tectonic and underlying bedrock. These factors may become secondary in catchments affected by glacial erosion, as it is the case in many mid- to high-latitude mountain belts. The perturbation (i.e. the destruction) of an initial steady state fluvial bedrock morphology (where uplift is balanced by surface lowering rates) will tend to become successively larger if the repeated action of glacial processes exceeds the potential of fluvial readjustment during deglaciated periods. Topographic change is associated with a decrease and fragmentation of the channel network and an extension of the hillslope domain. In case of glacially conditioned catchments discrimination of the two domains remains problematic and a discrimination inconsistent. A definition is therefore highly needed considering that (i) a spatial shift in the domains affect the process and rate of erosion and (ii) topographic classifications of alpine catchments often base on channel and hillslope parameters (i.e.channel or hillslope relief). Here we propose a novel numerical approach to topographically define channel heads from digital topography in glacially conditioned mountain range catchments in order to discriminate the channel from the hillslope domain. We analyzed the topography of the southern European Central Alps, a region which (i) has been glaciated multiple times during the Quaternary, shows (ii) little lithological variations, is (iii) home of very low erodible rocks and is (iv) known as a region were tectonic processes have largely ceased. The

Diverse requirements for microglial survival, specification, and function revealed by defined-medium cultures

PubMed Central

Bohlen, Christopher J.; Bennett, F. Chris; Tucker, Andrew F.; Collins, Hannah Y.; Mulinyawe, Sara B.; Barres, Ben A.

2017-01-01

Summary Microglia, the resident macrophages of the central nervous system (CNS), engage in various CNS-specific functions that are critical for development and health. To better study microglia and the properties that distinguish them from other tissue macrophage populations, we have optimized serum-free culture conditions to permit robust survival of highly ramified adult microglia under defined-medium conditions. We find that astrocyte-derived factors prevent microglial death ex vivo and that this activity results from three primary components, CSF-1/IL-34, TGF-β2, and cholesterol. Using microglial cultures that have never been exposed to serum, we demonstrate a dramatic and lasting change in phagocytic capacity after serum exposure. Finally, we find that mature microglia rapidly lose signature gene expression after isolation, and that this loss can be reversed by engrafting cells back into an intact CNS environment. These data indicate that the specialized gene expression profile of mature microglia requires continuous instructive signaling from the intact CNS. PMID:28521131

Diverse Requirements for Microglial Survival, Specification, and Function Revealed by Defined-Medium Cultures.

PubMed

Bohlen, Christopher J; Bennett, F Chris; Tucker, Andrew F; Collins, Hannah Y; Mulinyawe, Sara B; Barres, Ben A

2017-05-17

Microglia, the resident macrophages of the CNS, engage in various CNS-specific functions that are critical for development and health. To better study microglia and the properties that distinguish them from other tissue macrophage populations, we have optimized serum-free culture conditions to permit robust survival of highly ramified adult microglia under defined-medium conditions. We find that astrocyte-derived factors prevent microglial death ex vivo and that this activity results from three primary components, CSF-1/IL-34, TGF-β2, and cholesterol. Using microglial cultures that have never been exposed to serum, we demonstrate a dramatic and lasting change in phagocytic capacity after serum exposure. Finally, we find that mature microglia rapidly lose signature gene expression after isolation, and that this loss can be reversed by engrafting cells back into an intact CNS environment. These data indicate that the specialized gene expression profile of mature microglia requires continuous instructive signaling from the intact CNS. Copyright © 2017 Elsevier Inc. All rights reserved.

Serum-free light-chain analysis in diagnosis and management of multiple myeloma and related conditions.

PubMed

Milani, Paolo; Palladini, Giovanni; Merlini, Giampaolo

2016-01-01

The introduction of the serum-free light-chain (S-FLC) assay has been a breakthrough in the diagnosis and management of plasma cell dyscrasias, particularly monoclonal light-chain diseases. The first method, proposed in 2001, quantifies serum-free light-chains using polyclonal antibodies. More recently, assays based on monoclonal antibodies have entered into clinical practice. S-FLC measurement plays a central role in the screening for multiple myeloma and related conditions, in association with electrophoretic techniques. Analysis of S-FLC is essential in assessing the risk of progression of precursor diseases to overt plasma cell dyscrasias. It is also useful for risk stratification in solitary plasmacytoma and AL amyloidosis. The S-FLC measurement is part of the new diagnostic criteria for multiple myeloma, and provides a marker to follow changes in clonal substructure over time. Finally, the evaluation of S-FLC is fundamental for assessing the response to treatment in monoclonal light chain diseases.

Oil-Free Turbomachinery Team Passed Milestone on Path to the First Oil-Free Turbine Aircraft Engine

NASA Technical Reports Server (NTRS)

Bream, Bruce L.

2002-01-01

The Oil-Free Turbine Engine Technology Project team successfully demonstrated a foil-air bearing designed for the core rotor shaft of a turbine engine. The bearings were subjected to test conditions representative of the engine core environment through a combination of high speeds, sustained loads, and elevated temperatures. The operational test envelope was defined during conceptual design studies completed earlier this year by bearing manufacturer Mohawk Innovative Technologies and the turbine engine company Williams International. The prototype journal foil-air bearings were tested at the NASA Glenn Research Center. Glenn is working with Williams and Mohawk to create a revolution in turbomachinery by developing the world's first Oil-Free turbine aircraft engine. NASA's General Aviation Propulsion project and Williams International recently developed the FJX-2 turbofan engine that is being commercialized as the EJ-22. This core bearing milestone is a first step toward a future version of the EJ-22 that will take advantage of recent advances in foil-air bearings by eliminating the need for oil lubrication systems and rolling element bearings. Oil-Free technology can reduce engine weight by 15 percent and let engines operate at very high speeds, yielding power density improvements of 20 percent, and reducing engine maintenance costs. In addition, with NASA coating technology, engines can operate at temperatures up to 1200 F. Although the project is still a couple of years from a full engine test of the bearings, this milestone shows that the bearing design exceeds the expected environment, thus providing confidence that an Oil-Free turbine aircraft engine will be attained. The Oil-Free Turbomachinery Project is supported through the Aeropropulsion Base Research Program.

Fitbit Charge HR Wireless Heart Rate Monitor: Validation Study Conducted Under Free-Living Conditions.

PubMed

Gorny, Alexander Wilhelm; Liew, Seaw Jia; Tan, Chuen Seng; Müller-Riemenschneider, Falk

2017-10-20

Many modern smart watches and activity trackers feature an optical sensor that estimates the wearer's heart rate. Recent studies have evaluated the performance of these consumer devices in the laboratory. The objective of our study was to examine the accuracy and sensitivity of a common wrist-worn tracker device in measuring heart rates and detecting 1-min bouts of moderate to vigorous physical activity (MVPA) under free-living conditions. Ten healthy volunteers were recruited from a large university in Singapore to participate in a limited field test, followed by a month of continuous data collection. During the field test, each participant would wear one Fitbit Charge HR activity tracker and one Polar H6 heart rate monitor. Fitbit measures were accessed at 1-min intervals, while Polar readings were available for 10-s intervals. We derived intraclass correlation coefficients (ICCs) for individual participants comparing heart rate estimates. We applied Centers for Disease Control and Prevention heart rate zone cut-offs to ascertain the sensitivity and specificity of Fitbit in identifying 1-min epochs falling into MVPA heart rate zone. We collected paired heart rate data for 2509 1-min epochs in 10 individuals under free-living conditions of 3 to 6 hours. The overall ICC comparing 1-min Fitbit measures with average 10-s Polar H6 measures for the same epoch was .83 (95% CI .63-.91). On average, the Fitbit tracker underestimated heart rate measures by -5.96 bpm (standard error, SE=0.18). At the low intensity heart rate zone, the underestimate was smaller at -4.22 bpm (SE=0.15). This underestimate grew to -16.2 bpm (SE=0.74) in the MVPA heart rate zone. Fitbit devices detected 52.9% (192/363) of MVPA heart rate zone epochs correctly. Positive and negative predictive values were 86.1% (192/223) and 92.52% (2115/2286), respectively. During subsequent 1 month of continuous data collection (270 person-days), only 3.9% of 1-min epochs could be categorized as MVPA according

A numerical procedure for transient free surface seepage through fracture networks

NASA Astrophysics Data System (ADS)

Jiang, Qinghui; Ye, Zuyang; Zhou, Chuangbing

2014-11-01

A parabolic variational inequality (PVI) formulation is presented for the transient free surface seepage problem defined for a whole fracture network. Because the seepage faces are specified as Signorini-type conditions, the PVI formulation can effectively eliminate the singularity of spillpoints that evolve with time. By introducing a continuous penalty function to replace the original Heaviside function, a finite element procedure based on the PVI formulation is developed to predict the transient free surface response in the fracture network. The effects of the penalty parameter on the solution precision are analyzed. A relative error formula for evaluating the flow losses at steady state caused by the penalty parameter is obtained. To validate the proposed method, three typical examples are solved. The solutions for the first example are compared with the experimental results. The results from the last two examples further demonstrate that the orientation, extent and density of fractures significantly affect the free surface seepage behavior in the fracture network.

The valence-specific laterality effect in free viewing conditions: The influence of sex, handedness, and response bias.

PubMed

Rodway, Paul; Wright, Lynn; Hardie, Scott

2003-12-01

The right hemisphere has often been viewed as having a dominant role in the processing of emotional information. Other evidence indicates that both hemispheres process emotional information but their involvement is valence specific, with the right hemisphere dealing with negative emotions and the left hemisphere preferentially processing positive emotions. This has been found under both restricted (Reuter-Lorenz & Davidson, 1981) and free viewing conditions (Jansari, Tranel, & Adophs, 2000). It remains unclear whether the valence-specific laterality effect is also sex specific or is influenced by the handedness of participants. To explore this issue we repeated Jansari et al.'s free-viewing laterality task with 78 participants. We found a valence-specific laterality effect in women but not men, with women discriminating negative emotional expressions more accurately when the face was presented on the left-hand side and discriminating positive emotions more accurately when those faces were presented on the right-hand side. These results indicate that under free viewing conditions women are more lateralised for the processing of facial emotion than are men. Handedness did not affect the lateralised processing of facial emotion. Finally, participants demonstrated a response bias on control trials, where facial emotion did not differ between the faces. Participants selected the left-hand side more frequently when they believed the expression was negative and the right-hand side more frequently when they believed the expression was positive. This response bias can cause a spurious valence-specific laterality effect which might have contributed to the conflicting findings within the literature.

Thermocapillary convection in two immiscible liquid layers with free surface

NASA Technical Reports Server (NTRS)

Doi, Takao; Koster, Jean N.

1993-01-01

Thermocapillary convection is studied in two immiscible liquid layers with one free surface, one liquid/liquid interface, and differential heating applied parallel to the interfaces. An analytical solution is introduced for infinite horizontal layers. The defining parameter for the flow pattern is lambda, the ratio of the temperature coefficient of the interfacial tension to that of the surface tension. Four different flow patterns exist under zero gravity conditions. 'Halt' conditions which halt the fluid motion in the lower encapsulated liquid layer have been found. A numerical experiment is carried out to study effects of vertical end walls on the double layer convection in a 2D cavity. The halt condition obtained from the analytical study is found to be valid in the limit of small Reynolds numbers. The flow in the encapsulated liquid layer can be suppressed substantially.

An analytical study of the free and forced vibration response of a ribbed plate with free boundary conditions

NASA Astrophysics Data System (ADS)

Lin, Tian Ran; Zhang, Kai

2018-05-01

An analytical study to predict the vibration response of a ribbed plate with free boundary conditions is presented. The analytical solution was derived using a double cosine integral transform technique and then utilized to study the free and forced vibration of the ribbed plate, as well as the effect of the rib on the modal response of the uniform plate. It is shown that in addition to the three zero-frequency rigid body modes of the plate, the vibration modes of the uniform plate can be classified into four mode groups according to the symmetric properties of the plate with respect to the two orthogonal middle lines parallel to the plate edges. The four mode groups correspond to a double symmetric group, a double anti-symmetric group and two symmetric/anti-symmetric groups. Whilst the inclusion of the rib to the plate is shown to cause distortion to the distribution of vibration modes, most modes can still be traced back to the original modes of the uniform plate. Both the mass and stiffness of the rib are shown to affect the modal vibration of the uniform plate, whereby a dominant effect from the rib mass leads to a decrease in the modal frequency of the plate, whereas a dominant effect from the rib stiffness leads to an increase in plate modal frequency. When the stiffened rib behaves as an effective boundary to the plate vibration, an original plate mode becomes a pair of degenerate modes, whereby one mode has a higher frequency and the other mode has a lower frequency than that of the original mode.

Gödel, Tarski, Turing and the Conundrum of Free Will

NASA Astrophysics Data System (ADS)

Nayakar, Chetan S. Mandayam; Srikanth, R.

2014-07-01

The problem of defining and locating free will (FW) in physics is studied. On the basis of logical paradoxes, we argue that FW has a metatheoretic character, like the concept of truth in Tarski's undefinability theorem. Free will exists relative to a base theory if there is freedom to deviate from the deterministic or indeterministic dynamics in the theory, with the deviations caused by parameters (representing will) in the meta-theory. By contrast, determinism and indeterminism do not require meta-theoretic considerations in their formalization, making FW a fundamentally new causal primitive. FW exists relative to the meta-theory if there is freedom for deviation, due to higher-order causes. Absolute free will, which corresponds to our intuitive introspective notion of free will, exists if this meta-theoretic hierarchy is infinite. We argue that this hierarchy corresponds to higher levels of uncomputability. In other words, at any finitely high order in the hierarchy, there are uncomputable deviations from the law at that order. Applied to the human condition, the hierarchy corresponds to deeper levels of the subconscious or unconscious mind. Possible ramifications of our model for physics, neuroscience and artificial intelligence (AI) are briefly considered.

Chemoselective Hydrogenation of Aldehydes under Mild, Base-Free Conditions: Manganese Outperforms Rhenium

PubMed Central

2018-01-01

Several hydride Mn(I) and Re(I) PNP pincer complexes were applied as catalysts for the homogeneous chemoselective hydrogenation of aldehydes. Among these, [Mn(PNP-iPr)(CO)2(H)] was found to be one of the most efficient base metal catalysts for this process and represents a rare example which permits the selective hydrogenation of aldehydes in the presence of ketones and other reducible functionalities, such as C=C double bonds, esters, or nitriles. The reaction proceeds at room temperature under base-free conditions with catalyst loadings between 0.1 and 0.05 mol% and a hydrogen pressure of 50 bar (reaching TONs of up to 2000). A mechanism which involves an outer-sphere hydride transfer and reversible PNP ligand deprotonation/protonation is proposed. Analogous isoelectronic and isostructural Re(I) complexes were only poorly active. PMID:29755828

Controlling cell-free metabolism through physiochemical perturbations.

PubMed

Karim, Ashty S; Heggestad, Jacob T; Crowe, Samantha A; Jewett, Michael C

2018-01-01

Building biosynthetic pathways and engineering metabolic reactions in cells can be time-consuming due to complexities in cellular metabolism. These complexities often convolute the combinatorial testing of biosynthetic pathway designs needed to define an optimal biosynthetic system. To simplify the optimization of biosynthetic systems, we recently reported a new cell-free framework for pathway construction and testing. In this framework, multiple crude-cell extracts are selectively enriched with individual pathway enzymes, which are then mixed to construct full biosynthetic pathways on the time scale of a day. This rapid approach to building pathways aids in the study of metabolic pathway performance by providing a unique freedom of design to modify and control biological systems for both fundamental and applied biotechnology. The goal of this work was to demonstrate the ability to probe biosynthetic pathway performance in our cell-free framework by perturbing physiochemical conditions, using n-butanol synthesis as a model. We carried out three unique case studies. First, we demonstrated the power of our cell-free approach to maximize biosynthesis yields by mapping physiochemical landscapes using a robotic liquid-handler. This allowed us to determine that NAD and CoA are the most important factors that govern cell-free n-butanol metabolism. Second, we compared metabolic profile differences between two different approaches for building pathways from enriched lysates, heterologous expression and cell-free protein synthesis. We discover that phosphate from PEP utilization, along with other physiochemical reagents, during cell-free protein synthesis-coupled, crude-lysate metabolic system operation inhibits optimal cell-free n-butanol metabolism. Third, we show that non-phosphorylated secondary energy substrates can be used to fuel cell-free protein synthesis and n-butanol biosynthesis. Taken together, our work highlights the ease of using cell-free systems to explore

An extension to artifact-free projection overlaps

DOE Office of Scientific and Technical Information (OSTI.GOV)

Lin, Jianyu, E-mail: jianyulin@hotmail.com

2015-05-15

Purpose: In multipinhole single photon emission computed tomography, the overlapping of projections has been used to increase sensitivity. Avoiding artifacts in the reconstructed image associated with projection overlaps (multiplexing) is a critical issue. In our previous report, two types of artifact-free projection overlaps, i.e., projection overlaps that do not lead to artifacts in the reconstructed image, were formally defined and proved, and were validated via simulations. In this work, a new proposition is introduced to extend the previously defined type-II artifact-free projection overlaps so that a broader range of artifact-free overlaps is accommodated. One practical purpose of the new extensionmore » is to design a baffle window multipinhole system with artifact-free projection overlaps. Methods: First, the extended type-II artifact-free overlap was theoretically defined and proved. The new proposition accommodates the situation where the extended type-II artifact-free projection overlaps can be produced with incorrectly reconstructed portions in the reconstructed image. Next, to validate the theory, the extended-type-II artifact-free overlaps were employed in designing the multiplexing multipinhole spiral orbit imaging systems with a baffle window. Numerical validations were performed via simulations, where the corresponding 1-pinhole nonmultiplexing reconstruction results were used as the benchmark for artifact-free reconstructions. The mean square error (MSE) was the metric used for comparisons of noise-free reconstructed images. Noisy reconstructions were also performed as part of the validations. Results: Simulation results show that for noise-free reconstructions, the MSEs of the reconstructed images of the artifact-free multiplexing systems are very similar to those of the corresponding 1-pinhole systems. No artifacts were observed in the reconstructed images. Therefore, the testing results for artifact-free multiplexing systems designed using

User-defined Material Model for Thermo-mechanical Progressive Failure Analysis

NASA Technical Reports Server (NTRS)

Knight, Norman F., Jr.

2008-01-01

Previously a user-defined material model for orthotropic bimodulus materials was developed for linear and nonlinear stress analysis of composite structures using either shell or solid finite elements within a nonlinear finite element analysis tool. Extensions of this user-defined material model to thermo-mechanical progressive failure analysis are described, and the required input data are documented. The extensions include providing for temperature-dependent material properties, archival of the elastic strains, and a thermal strain calculation for materials exhibiting a stress-free temperature.

Solvent-free synthesis

EPA Science Inventory

This chapter gives a brief introduction about solvent-free reactions whose importance can be gauged by the increasing number of publications every year during the last decade. The mechanistic aspects of the reactions under solvent-free conditions have been highlighted. Our observ...

Cryopreservation of Human Pluripotent Stem Cells in Defined Medium

PubMed Central

Liu, Weiwei; Chen, Guokai

2014-01-01

This protocol describes a cryopreservation procedure using an enzyme-free dissociation method to harvest cells and preserve cells in albumin-free chemically defined E8 medium for human pluripotent stem cells (hPSCs). The dissociation by EDTA/PBS produces small cell aggregates that allow high survival efficiency in passaging and cryopreservation. The preservation in E8 medium eliminates serum or other animal products, and is suitable for the increasing demand for high quality hPSCs in translational research. In combination with the special feature of EDTA/PBS dissociation, this protocol allows efficient cryopreservation in more time-saving manner. PMID:25366897

Patterned graphene functionalization via mask-free scanning of micro-plasma jet under ambient condition

NASA Astrophysics Data System (ADS)

Ye, Dong; Wu, Shu-Qun; Yu, Yao; Liu, Lin; Lu, Xin-Pei; Wu, Yue

2014-03-01

In this work, a mask-free method is introduced for patterned nitrogen doping of graphene using a micro-plasma jet under ambient condition. Raman and X-ray photoelectron spectroscopy spectra indicate that nitrogen atoms are incorporated into the graphene lattice with the two-dimensional spatial distribution precisely controlled in the range of mm down to 10 μm. Since the chemistry of the micro-plasma jet can be controlled by the choice of the gas mixture, this direct writing process with micro-plasma jet can be a versatile approach for patterned functionalization of graphene with high spatial resolution. This could have promising applications in graphene-based electronics.

The eMouveRecherche application competes with research devices to evaluate energy expenditure, physical activity and still time in free-living conditions.

PubMed

Guidoux, Romain; Duclos, Martine; Fleury, Gérard; Lacomme, Philippe; Lamaudière, Nicolas; Saboul, Damien; Ren, Libo; Rousset, Sylvie

2017-05-01

The proliferation of smartphones is creating new opportunities to monitor and interact with human subjects in free-living conditions since smartphones are familiar to large segments of the population and facilitate data collection, transmission and analysis. From accelerometry data collected by smartphones, the present work aims to estimate time spent in different activity categories and the energy expenditure in free-living conditions. Our research encompasses the definition of an energy-saving function (Pred EE ) considering four physical categories of activities (still, light, moderate and vigorous), their duration and metabolic cost (MET). To create an efficient discrimination function, the method consists of classifying accelerometry-transformed signals into categories and of associating each category with corresponding Metabolic Equivalent Tasks. The performance of the Pred EE function was compared with two previously published functions (f(η,d)aedes,f(η,d)nrjsi), and with two dedicated sensors (Armband® and Actiheart®) in free-living conditions over a 12-h monitoring period using 30 volunteers. Compared to the two previous functions, Pred EE was the only one able to provide estimations of time spent in each activity category. In relative value, all the activity categories were evaluated similarly to those given by Armband®. Compared to Actiheart®, the function underestimated still activities by 10.1% and overestimated light- and moderate-intensity activities by 7.9% and 4.2%, respectively. The total energy expenditure error produced by Pred EE compared to Armband® was lower than those given by the two previous functions (5.7% vs. 14.1% and 17.0%). Pred EE provides the user with an accurate physical activity feedback which should help self-monitoring in free-living conditions. Copyright © 2017 Elsevier Inc. All rights reserved.

Determination of Membrane-Insertion Free Energies by Molecular Dynamics Simulations

PubMed Central

Gumbart, James; Roux, Benoît

2012-01-01

The accurate prediction of membrane-insertion probability for arbitrary protein sequences is a critical challenge to identifying membrane proteins and determining their folded structures. Although algorithms based on sequence statistics have had moderate success, a complete understanding of the energetic factors that drive the insertion of membrane proteins is essential to thoroughly meeting this challenge. In the last few years, numerous attempts to define a free-energy scale for amino-acid insertion have been made, yet disagreement between most experimental and theoretical scales persists. However, for a recently resolved water-to-bilayer scale, it is found that molecular dynamics simulations that carefully mimic the conditions of the experiment can reproduce experimental free energies, even when using the same force field as previous computational studies that were cited as evidence of this disagreement. Therefore, it is suggested that experimental and simulation-based scales can both be accurate and that discrepancies stem from disparities in the microscopic processes being considered rather than methodological errors. Furthermore, these disparities make the development of a single universally applicable membrane-insertion free energy scale difficult. PMID:22385850

Observer efficiency in free-localization tasks with correlated noise.

PubMed

Abbey, Craig K; Eckstein, Miguel P

2014-01-01

The efficiency of visual tasks involving localization has traditionally been evaluated using forced choice experiments that capitalize on independence across locations to simplify the performance of the ideal observer. However, developments in ideal observer analysis have shown how an ideal observer can be defined for free-localization tasks, where a target can appear anywhere in a defined search region and subjects respond by localizing the target. Since these tasks are representative of many real-world search tasks, it is of interest to evaluate the efficiency of observer performance in them. The central question of this work is whether humans are able to effectively use the information in a free-localization task relative to a similar task where target location is fixed. We use a yes-no detection task at a cued location as the reference for this comparison. Each of the tasks is evaluated using a Gaussian target profile embedded in four different Gaussian noise backgrounds having power-law noise power spectra with exponents ranging from 0 to 3. The free localization task had a square 6.7° search region. We report on two follow-up studies investigating efficiency in a detect-and-localize task, and the effect of processing the white-noise backgrounds. In the fixed-location detection task, we find average observer efficiency ranges from 35 to 59% for the different noise backgrounds. Observer efficiency improves dramatically in the tasks involving localization, ranging from 63 to 82% in the forced localization tasks and from 78 to 92% in the detect-and- localize tasks. Performance in white noise, the lowest efficiency condition, was improved by filtering to give them a power-law exponent of 2. Classification images, used to examine spatial frequency weights for the tasks, show better tuning to ideal weights in the free-localization tasks. The high absolute levels of efficiency suggest that observers are well-adapted to free-localization tasks.

Observer efficiency in free-localization tasks with correlated noise

PubMed Central

Abbey, Craig K.; Eckstein, Miguel P.

2014-01-01

The efficiency of visual tasks involving localization has traditionally been evaluated using forced choice experiments that capitalize on independence across locations to simplify the performance of the ideal observer. However, developments in ideal observer analysis have shown how an ideal observer can be defined for free-localization tasks, where a target can appear anywhere in a defined search region and subjects respond by localizing the target. Since these tasks are representative of many real-world search tasks, it is of interest to evaluate the efficiency of observer performance in them. The central question of this work is whether humans are able to effectively use the information in a free-localization task relative to a similar task where target location is fixed. We use a yes-no detection task at a cued location as the reference for this comparison. Each of the tasks is evaluated using a Gaussian target profile embedded in four different Gaussian noise backgrounds having power-law noise power spectra with exponents ranging from 0 to 3. The free localization task had a square 6.7° search region. We report on two follow-up studies investigating efficiency in a detect-and-localize task, and the effect of processing the white-noise backgrounds. In the fixed-location detection task, we find average observer efficiency ranges from 35 to 59% for the different noise backgrounds. Observer efficiency improves dramatically in the tasks involving localization, ranging from 63 to 82% in the forced localization tasks and from 78 to 92% in the detect-and- localize tasks. Performance in white noise, the lowest efficiency condition, was improved by filtering to give them a power-law exponent of 2. Classification images, used to examine spatial frequency weights for the tasks, show better tuning to ideal weights in the free-localization tasks. The high absolute levels of efficiency suggest that observers are well-adapted to free-localization tasks. PMID:24817854

Defined PEG smears as an alternative approach to enhance the search for crystallization conditions and crystal-quality improvement in reduced screens

DOE Office of Scientific and Technical Information (OSTI.GOV)

Chaikuad, Apirat, E-mail: apirat.chaikuad@sgc.ox.ac.uk; Knapp, Stefan; Johann Wolfgang Goethe-University, Building N240 Room 3.03, Max-von-Laue-Strasse 9, 60438 Frankfurt am Main

An alternative strategy for PEG sampling is suggested through the use of four newly defined PEG smears to enhance chemical space in reduced screens with a benefit towards protein crystallization. The quest for an optimal limited set of effective crystallization conditions remains a challenge in macromolecular crystallography, an issue that is complicated by the large number of chemicals which have been deemed to be suitable for promoting crystal growth. The lack of rational approaches towards the selection of successful chemical space and representative combinations has led to significant overlapping conditions, which are currently present in a multitude of commercially availablemore » crystallization screens. Here, an alternative approach to the sampling of widely used PEG precipitants is suggested through the use of PEG smears, which are mixtures of different PEGs with a requirement of either neutral or cooperatively positive effects of each component on crystal growth. Four newly defined smears were classified by molecular-weight groups and enabled the preservation of specific properties related to different polymer sizes. These smears not only allowed a wide coverage of properties of these polymers, but also reduced PEG variables, enabling greater sampling of other parameters such as buffers and additives. The efficiency of the smear-based screens was evaluated on more than 220 diverse recombinant human proteins, which overall revealed a good initial crystallization success rate of nearly 50%. In addition, in several cases successful crystallizations were only obtained using PEG smears, while various commercial screens failed to yield crystals. The defined smears therefore offer an alternative approach towards PEG sampling, which will benefit the design of crystallization screens sampling a wide chemical space of this key precipitant.« less

Control of flexible beams using a free-free active truss

NASA Technical Reports Server (NTRS)

Clark, W. W.; Kimiavi, B.; Robertshaw, H. H.

1989-01-01

An analytical and experimental study involving controlling flexible beams using a free-free active truss is presented. This work extends previous work in controlling flexible continua with active trusses which were configured with fixed-free boundary conditions. The following describes the Lagrangian approach used to derive the equations of motion for the active truss and the beams attached to it. A partial-state feedback control law is derived for this system based on a full-state feedback Linear Quadratic Regulator method. The analytical model is examined via numerical simulations and the results are compared to a similar experimental apparatus described herein. The results show that control of a flexible continua is possible with a free-free active truss.

Spin trapping combined with quantitative mass spectrometry defines free radical redistribution within the oxidized hemoglobin:haptoglobin complex.

PubMed

Vallelian, Florence; Garcia-Rubio, Ines; Puglia, Michele; Kahraman, Abdullah; Deuel, Jeremy W; Engelsberger, Wolfgang R; Mason, Ronald P; Buehler, Paul W; Schaer, Dominik J

2015-08-01

Extracellular or free hemoglobin (Hb) accumulates during hemolysis, tissue damage, and inflammation. Heme-triggered oxidative reactions can lead to diverse structural modifications of lipids and proteins, which contribute to the propagation of tissue damage. One important target of Hb׳s peroxidase reactivity is its own globin structure. Amino acid oxidation and crosslinking events destabilize the protein and ultimately cause accumulation of proinflammatory and cytotoxic Hb degradation products. The Hb scavenger haptoglobin (Hp) attenuates oxidation-induced Hb degradation. In this study we show that in the presence of hydrogen peroxide (H2O2), Hb and the Hb:Hp complex share comparable peroxidative reactivity and free radical generation. While oxidation of both free Hb and Hb:Hp complex generates a common tyrosine-based free radical, the spin-trapping reaction with 5,5-dimethyl-1-pyrroline N-oxide (DMPO) yields dissimilar paramagnetic products in Hb and Hb:Hp, suggesting that radicals are differently redistributed within the complex before reacting with the spin trap. With LC-MS(2) mass spectrometry we assigned multiple known and novel DMPO adduct sites. Quantification of these adducts suggested that the Hb:Hp complex formation causes extensive delocalization of accessible free radicals with drastic reduction of the major tryptophan and cysteine modifications in the β-globin chain of the Hb:Hp complex, including decreased βCys93 DMPO adduction. In contrast, the quantitative changes in DMPO adduct formation on Hb:Hp complex formation were less pronounced in the Hb α-globin chain. In contrast to earlier speculations, we found no evidence that free Hb radicals are delocalized to the Hp chain of the complex. The observation that Hb:Hp complex formation alters free radical distribution in Hb may help to better understand the structural basis for Hp as an antioxidant protein. Copyright © 2015 Elsevier Inc. All rights reserved.

29 CFR 779.505 - “Oppressive child labor” defined.

Code of Federal Regulations, 2014 CFR

2014-07-01

... 29 Labor 3 2014-07-01 2014-07-01 false âOppressive child laborâ defined. 779.505 Section 779.505... Child Labor Provisions § 779.505 “Oppressive child labor” defined. Section 3(1) of the Act defines oppressive child labor as follows: “Oppressive child labor” means a condition of employment under which (1...

29 CFR 779.505 - “Oppressive child labor” defined.

Code of Federal Regulations, 2010 CFR

2010-07-01

... 29 Labor 3 2010-07-01 2010-07-01 false âOppressive child laborâ defined. 779.505 Section 779.505... Child Labor Provisions § 779.505 “Oppressive child labor” defined. Section 3(1) of the Act defines oppressive child labor as follows: “Oppressive child labor” means a condition of employment under which (1...

29 CFR 779.505 - “Oppressive child labor” defined.

Code of Federal Regulations, 2011 CFR

2011-07-01

... 29 Labor 3 2011-07-01 2011-07-01 false âOppressive child laborâ defined. 779.505 Section 779.505... Child Labor Provisions § 779.505 “Oppressive child labor” defined. Section 3(1) of the Act defines oppressive child labor as follows: “Oppressive child labor” means a condition of employment under which (1...

29 CFR 779.505 - “Oppressive child labor” defined.

Code of Federal Regulations, 2012 CFR

2012-07-01

... 29 Labor 3 2012-07-01 2012-07-01 false âOppressive child laborâ defined. 779.505 Section 779.505... Child Labor Provisions § 779.505 “Oppressive child labor” defined. Section 3(1) of the Act defines oppressive child labor as follows: “Oppressive child labor” means a condition of employment under which (1...

29 CFR 779.505 - “Oppressive child labor” defined.

Code of Federal Regulations, 2013 CFR

2013-07-01

... 29 Labor 3 2013-07-01 2013-07-01 false âOppressive child laborâ defined. 779.505 Section 779.505... Child Labor Provisions § 779.505 “Oppressive child labor” defined. Section 3(1) of the Act defines oppressive child labor as follows: “Oppressive child labor” means a condition of employment under which (1...

Defining a region of optimization based on engine usage data

DOEpatents

Jiang, Li; Lee, Donghoon; Yilmaz, Hakan; Stefanopoulou, Anna

2015-08-04

Methods and systems for engine control optimization are provided. One or more operating conditions of a vehicle engine are detected. A value for each of a plurality of engine control parameters is determined based on the detected one or more operating conditions of the vehicle engine. A range of the most commonly detected operating conditions of the vehicle engine is identified and a region of optimization is defined based on the range of the most commonly detected operating conditions of the vehicle engine. The engine control optimization routine is initiated when the one or more operating conditions of the vehicle engine are within the defined region of optimization.

Two-dimensional grid-free compressive beamforming.

PubMed

Yang, Yang; Chu, Zhigang; Xu, Zhongming; Ping, Guoli

2017-08-01

Compressive beamforming realizes the direction-of-arrival (DOA) estimation and strength quantification of acoustic sources by solving an underdetermined system of equations relating microphone pressures to a source distribution via compressive sensing. The conventional method assumes DOAs of sources to lie on a grid. Its performance degrades due to basis mismatch when the assumption is not satisfied. To overcome this limitation for the measurement with plane microphone arrays, a two-dimensional grid-free compressive beamforming is developed. First, a continuum based atomic norm minimization is defined to denoise the measured pressure and thus obtain the pressure from sources. Next, a positive semidefinite programming is formulated to approximate the atomic norm minimization. Subsequently, a reasonably fast algorithm based on alternating direction method of multipliers is presented to solve the positive semidefinite programming. Finally, the matrix enhancement and matrix pencil method is introduced to process the obtained pressure and reconstruct the source distribution. Both simulations and experiments demonstrate that under certain conditions, the grid-free compressive beamforming can provide high-resolution and low-contamination imaging, allowing accurate and fast estimation of two-dimensional DOAs and quantification of source strengths, even with non-uniform arrays and noisy measurements.

A semi-implicit augmented IIM for Navier–Stokes equations with open, traction, or free boundary conditions

PubMed Central

Li, Zhilin; Xiao, Li; Cai, Qin; Zhao, Hongkai; Luo, Ray

2016-01-01

In this paper, a new Navier–Stokes solver based on a finite difference approximation is proposed to solve incompressible flows on irregular domains with open, traction, and free boundary conditions, which can be applied to simulations of fluid structure interaction, implicit solvent model for biomolecular applications and other free boundary or interface problems. For some problems of this type, the projection method and the augmented immersed interface method (IIM) do not work well or does not work at all. The proposed new Navier–Stokes solver is based on the local pressure boundary method, and a semi-implicit augmented IIM. A fast Poisson solver can be used in our algorithm which gives us the potential for developing fast overall solvers in the future. The time discretization is based on a second order multi-step method. Numerical tests with exact solutions are presented to validate the accuracy of the method. Application to fluid structure interaction between an incompressible fluid and a compressible gas bubble is also presented. PMID:27087702

A semi-implicit augmented IIM for Navier-Stokes equations with open, traction, or free boundary conditions.

PubMed

Li, Zhilin; Xiao, Li; Cai, Qin; Zhao, Hongkai; Luo, Ray

2015-08-15

In this paper, a new Navier-Stokes solver based on a finite difference approximation is proposed to solve incompressible flows on irregular domains with open, traction, and free boundary conditions, which can be applied to simulations of fluid structure interaction, implicit solvent model for biomolecular applications and other free boundary or interface problems. For some problems of this type, the projection method and the augmented immersed interface method (IIM) do not work well or does not work at all. The proposed new Navier-Stokes solver is based on the local pressure boundary method, and a semi-implicit augmented IIM. A fast Poisson solver can be used in our algorithm which gives us the potential for developing fast overall solvers in the future. The time discretization is based on a second order multi-step method. Numerical tests with exact solutions are presented to validate the accuracy of the method. Application to fluid structure interaction between an incompressible fluid and a compressible gas bubble is also presented.

Towards a microscopic description of the free-energy landscape of water.

PubMed

Prada-Gracia, Diego; Shevchuk, Roman; Hamm, Peter; Rao, Francesco

2012-10-14

Free-energy landscape theory is often used to describe complex molecular systems. Here, a microscopic description of water structure and dynamics based on configuration-space-networks and molecular dynamics simulations of the TIP4P/2005 model is applied to investigate the free-energy landscape of water. The latter is built on top of a large set of water microstates describing the kinetic stability of local hydrogen-bond arrangements up to the second solvation shell. In temperature space, the landscape displays three different regimes. At around ambient conditions, the free-energy surface is characterized by many short-lived basins of attraction which are structurally well-defined (inhomogeneous regime). At lower temperatures instead, the liquid rapidly becomes homogeneous. In this regime, the free energy is funneled-like, with fully coordinated water arrangements at the bottom of the funnel. Finally, a third regime develops below the temperature of maximal compressibility (Widom line) where the funnel becomes steeper with few interconversions between microstates other than the fully coordinated ones. Our results present a way to manage the complexity of water structure and dynamics, connecting microscopic properties to its ensemble behavior.

Using Equation-Free Computation to Accelerate Network-Free Stochastic Simulation of Chemical Kinetics.

PubMed

Lin, Yen Ting; Chylek, Lily A; Lemons, Nathan W; Hlavacek, William S

2018-06-21

The chemical kinetics of many complex systems can be concisely represented by reaction rules, which can be used to generate reaction events via a kinetic Monte Carlo method that has been termed network-free simulation. Here, we demonstrate accelerated network-free simulation through a novel approach to equation-free computation. In this process, variables are introduced that approximately capture system state. Derivatives of these variables are estimated using short bursts of exact stochastic simulation and finite differencing. The variables are then projected forward in time via a numerical integration scheme, after which a new exact stochastic simulation is initialized and the whole process repeats. The projection step increases efficiency by bypassing the firing of numerous individual reaction events. As we show, the projected variables may be defined as populations of building blocks of chemical species. The maximal number of connected molecules included in these building blocks determines the degree of approximation. Equation-free acceleration of network-free simulation is found to be both accurate and efficient.

Hospital free cash flow.

PubMed

Kauer, R T; Silvers, J B

1991-01-01

Hospital managers may find it difficult to admit their investments have been suboptimal, but such investments often lead to poor returns and less future cash. Inappropriate use of free cash flow produces large transaction costs of exit. The relative efficiency of investor-owned and tax-exempt hospitals in the product market for hospital services is examined as the free cash flow theory is used to explore capital-market conditions of hospitals. Hypotheses concerning the current competitive conditions in the industry are set forth, and the implications of free cash flow for risk, capital-market efficiency, and the cost of capital to tax-exempt institution is compared to capital-market norms.

Defined three-dimensional culture conditions mediate efficient induction of definitive endoderm lineage from human umbilical cord Wharton's jelly mesenchymal stem cells.

PubMed

Al Madhoun, Ashraf; Ali, Hamad; AlKandari, Sarah; Atizado, Valerie Lopez; Akhter, Nadeem; Al-Mulla, Fahd; Atari, Maher

2016-11-16

Wharton's jelly-derived mesenchymal stem cells (WJ-MSCs) are gaining increasing interest as an alternative source of stem cells for regenerative medicine applications. Definitive endoderm (DE) specification is a prerequisite for the development of vital organs such as liver and pancreas. Hence, efficient induction of the DE lineage from stem cells is crucial for subsequent generation of clinically relevant cell types. Here we present a defined 3D differentiation protocol of WJ-MSCs into DE cells. WJ-MSCs were cultured in suspension to generate spheroids, about 1500 cells each, for 7 days. The serum-free differentiation media contained specific growth factors, cytokines, and small molecules that specifically regulate signaling pathways including sonic hedgehog, bone morphogenetic protein, Activin/Wnt, and Notch. We obtained more than 85 % DE cells as shown with FACS analysis using antibodies directed against the DE marker CXCR4. In addition, biochemical and molecular analysis of bona-fide DE markers revealed a time-course induction of Sox17, CXCR4, and FoxA2. Focused PCR-based array also indicated a specific induction into the DE lineage. In this study, we report an efficient serum-free protocol to differentiate WJ-MSCs into DE cells utilizing 3D spheroid formation. Our approach might aid in the development of new protocols to obtain DE-derivative lineages including liver-like and pancreatic insulin-producing cells.

Defining chaos.

PubMed

Hunt, Brian R; Ott, Edward

2015-09-01

In this paper, we propose, discuss, and illustrate a computationally feasible definition of chaos which can be applied very generally to situations that are commonly encountered, including attractors, repellers, and non-periodically forced systems. This definition is based on an entropy-like quantity, which we call "expansion entropy," and we define chaos as occurring when this quantity is positive. We relate and compare expansion entropy to the well-known concept of topological entropy to which it is equivalent under appropriate conditions. We also present example illustrations, discuss computational implementations, and point out issues arising from attempts at giving definitions of chaos that are not entropy-based.

Analysis and experimental study on formation conditions of large-scale barrier-free diffuse atmospheric pressure air plasmas in repetitive pulse mode

NASA Astrophysics Data System (ADS)

Li, Lee; Liu, Lun; Liu, Yun-Long; Bin, Yu; Ge, Ya-Feng; Lin, Fo-Chang

2014-01-01

Atmospheric air diffuse plasmas have enormous application potential in various fields of science and technology. Without dielectric barrier, generating large-scale air diffuse plasmas is always a challenging issue. This paper discusses and analyses the formation mechanism of cold homogenous plasma. It is proposed that generating stable diffuse atmospheric plasmas in open air should meet the three conditions: high transient power with low average power, excitation in low average E-field with locally high E-field region, and multiple overlapping electron avalanches. Accordingly, an experimental configuration of generating large-scale barrier-free diffuse air plasmas is designed. Based on runaway electron theory, a low duty-ratio, high voltage repetitive nanosecond pulse generator is chosen as a discharge excitation source. Using the wire-electrodes with small curvature radius, the gaps with highly non-uniform E-field are structured. Experimental results show that the volume-scaleable, barrier-free, homogeneous air non-thermal plasmas have been obtained between the gap spacing with the copper-wire electrodes. The area of air cold plasmas has been up to hundreds of square centimeters. The proposed formation conditions of large-scale barrier-free diffuse air plasmas are proved to be reasonable and feasible.

Defining Medical Capabilities for Exploration Missions

NASA Technical Reports Server (NTRS)

Hailey, M.; Antonsen, E.; Blue, R.; Reyes, D.; Mulcahy, R.; Kerstman, E.; Bayuse, T.

2018-01-01

Exploration-class missions to the moon, Mars and beyond will require a significant change in medical capability from today's low earth orbit centric paradigm. Significant increases in autonomy will be required due to differences in duration, distance and orbital mechanics. Aerospace medicine and systems engineering teams are working together within ExMC to meet these challenges. Identifying exploration medical system needs requires accounting for planned and unplanned medical care as defined in the concept of operations. In 2017, the ExMC Clinicians group identified medical capabilities to feed into the Systems Engineering process, including: determining what and how to address planned and preventive medical care; defining an Accepted Medical Condition List (AMCL) of conditions that may occur and a subset of those that can be treated effectively within the exploration environment; and listing the medical capabilities needed to treat those conditions in the AMCL. This presentation will discuss the team's approach to addressing these issues, as well as how the outputs of the clinical process impact the systems engineering effort.

Physical Activity Assessment Between Consumer- and Research-Grade Accelerometers: A Comparative Study in Free-Living Conditions.

PubMed

Dominick, Gregory M; Winfree, Kyle N; Pohlig, Ryan T; Papas, Mia A

2016-09-19

Wearable activity monitors such as Fitbit enable users to track various attributes of their physical activity (PA) over time and have the potential to be used in research to promote and measure PA behavior. However, the measurement accuracy of Fitbit in absolute free-living conditions is largely unknown. To examine the measurement congruence between Fitbit Flex and ActiGraph GT3X for quantifying steps, metabolic equivalent tasks (METs), and proportion of time in sedentary activity and light-, moderate-, and vigorous-intensity PA in healthy adults in free-living conditions. A convenience sample of 19 participants (4 men and 15 women), aged 18-37 years, concurrently wore the Fitbit Flex (wrist) and ActiGraph GT3X (waist) for 1- or 2-week observation periods (n=3 and n=16, respectively) that included self-reported bouts of daily exercise. Data were examined for daily activity, averaged over 14 days and for minutes of reported exercise. Average day-level data included steps, METs, and proportion of time in different intensity levels. Minute-level data included steps, METs, and mean intensity score (0 = sedentary, 3 = vigorous) for overall reported exercise bouts (N=120) and by exercise type (walking, n=16; run or sports, n=44; cardio machine, n=20). Measures of steps were similar between devices for average day- and minute-level observations (all P values > .05). Fitbit significantly overestimated METs for average daily activity, for overall minutes of reported exercise bouts, and for walking and run or sports exercises (mean difference 0.70, 1.80, 3.16, and 2.00 METs, respectively; all P values < .001). For average daily activity, Fitbit significantly underestimated the proportion of time in sedentary and light intensity by 20% and 34%, respectively, and overestimated time by 3% in both moderate and vigorous intensity (all P values < .001). Mean intensity scores were not different for overall minutes of exercise or for run or sports and cardio-machine exercises (all P

Enamel and dentin bond strengths of a new self-etch adhesive system.

PubMed

Walter, Ricardo; Swift, Edward J; Boushell, Lee W; Braswell, Krista

2011-12-01

statement of problem:  Self-etch adhesives typically are mildly acidic and therefore less effective than etch-and-rinse adhesives for bonding to enamel.   The purpose of this study was to evaluate the enamel and dentin shear bond strengths of a new two-step self-etch adhesive system, OptiBond XTR (Kerr Corporation, Orange, CA, USA).   The labial surfaces of 80 bovine teeth were ground to create flat, 600-grit enamel or dentin surfaces. Composite was bonded to enamel or dentin using the new two-step self-etch system or a three-step etch-and-rinse (OptiBond FL, Kerr), two-step self-etch (Clearfil SE Bond, Kuraray America, Houston, TX, USA), or one-step self-etch adhesive (Xeno IV, Dentsply Caulk, Milford, DE, USA). Following storage in water for 24 hours, shear bond strengths were determined using a universal testing machine. The enamel and dentin data sets were subjected to separate analysis of variance and Tukey's tests. Scanning electron microscopy was used to evaluate the effects of each system on enamel.   Mean shear bond strengths to enamel ranged from 18.1 MPa for Xeno IV to 41.0 MPa for OptiBond FL. On dentin, the means ranged from 33.3 MPa for OptiBond FL to 47.1 MPa for Clearfil SE Bond. OptiBond XTR performed as well as Clearfil SE Bond on dentin and as well as OptiBond FL on enamel. Field emission scanning electron microscope revealed that OptiBond XTR produced an enamel etch pattern that was less defined than that of OptiBond FL (37.5% phosphoric acid) but more defined than that of Clearfil SE Bond or Xeno IV.   The new two-step self-etch adhesive system formed excellent bonds to enamel and dentin in vitro. OptiBond XTR, a new two-step self-etch adhesive system, is a promising material for bonding to enamel as well as to dentin. © 2011 Wiley Periodicals, Inc.

Functionalized SBA-15 supported nickel (II)–oxime–imine catalysts for liquid phase oxidation of olefins under solvent-free conditions

DOE Office of Scientific and Technical Information (OSTI.GOV)

Paul, Luna; Banerjee, Biplab; Bhaumik, Asim, E-mail: msab@iacs.res.in

2016-05-15

A new oxime–imine functionalized highly ordered mesoporous SBA-15 (SBA-15-NH{sub 2}-DAMO) has been synthesized via post-synthesis functionalization of SBA-15 with 3-aminopropyl-triethoxysilane followed by the Schiff base condensation with diacetylmonooxime, which was further reacted with Ni(ClO{sub 4}){sub 2} to yield the functionalized nickel catalyst SBA-15-NH{sub 2}-DAMO-Ni. All the synthesized materials were thoroughly characterized using different characterization techniques. It was found that SBA-15-NH{sub 2}-DAMO-Ni catalyzes the one-pot oxidation of olefins like styrene, cyclohexene, cyclooctene, 1-hexene and 1-octene to the corresponding benzaldehyde, cyclohexene-1-ol and cyclooctene-oxide, respectively under solvent-free conditions by using tert-butylhydroperoxide as oxidant. - Graphical abstract: A new well characterized oxime–imine functionalized highlymore » ordered mesoporous SBA-15-NH{sub 2}-DAMO-Ni complex catalyzes the one-pot oxidation of olefins under solvent free mild conditions.« less

Successful Consecutive Expansion of Limbal Explants Using a Biosafe Culture Medium under Feeder Layer-Free Conditions.

PubMed

López-Paniagua, Marina; Nieto-Miguel, Teresa; de la Mata, Ana; Galindo, Sara; Herreras, José M; Corrales, Rosa M; Calonge, Margarita

2017-05-01

Transplantation of in vitro cultured limbal epithelial stem cells (LESCs) is a treatment widely used for LESC deficiency. However, the number of limbal tissue donors is limited, and protocols for LESC cultivation often include compounds and/or feeder layers that can induce side effects and/or increase the cost of the culture procedure. We investigated the feasibility of obtaining more than one limbal primary culture (LPC) from the same biopsy using a culture medium in which several potentially harmful compounds were replaced at the same time by biosafe supplements, allowing the LESC cultivation without feeder layers. We established feeder layer-free LPCs with three culture media: (1) a modified supplemental hormonal epithelial medium, containing potential harmful components (cholera toxin, dimethylsulfoxide, and fetal bovine serum [FBS]), (2) IOBA-FBS, a medium with FBS but with no other harmful supplements, and (3) IOBA-HS, similar to IOBA-FBS but with human serum instead of FBS. Additionally, the same limbal explant was consecutively cultured with IOBA-HS producing three cultures. LPCs were characterized by real-time reverse transcription polymerase chain reaction and/or immunofluorescence. LPCs cultured with the three media under feeder layer-free conditions showed cuboidal cells and no significant differences in the percentage of positive cells for limbal (ABCG2, p63, and K14) and corneal (K3, K12) proteins. Except for ABCG2, the relative mRNA expression of the LESC markers was significantly higher when IOBA-FBS or IOBA-HS was used. LPC1 showed characteristics similar to LPC0, while LPC2 cell morphology became elongated and the expression of some LESC markers was diminished. IOBA-HS enables the culturing of up to two biosafe homologous LPCs from one limbal tissue under feeder layer-free conditions. The routine use of this culture medium could improve both the biosafety and the number of available LPCs for potential clinical transplantation, as well as decrease

Preference for safflower oil in rats exposed to a cold environment under free-feeding conditions.

PubMed

Saitoh, Masaji; Ishii, Toshiaki; Takewaki, Tadashi; Nishimura, Masakazu

2005-07-01

There are several benefits to a high-fat diet for animals exposed to cold, including improved tolerance to severe cold conditions and increased survival rates in cold environments. It is therefore of interest to examine whether animals exposed to cold will selectively consume lipids. We examined the intake of safflower oil (SO) by rats exposed to cold (4 +/- 2 degrees C) under a feeding condition in which the rats were given free access to SO. Rats exposed to cold consumed more SO than those housed at 25 +/- 2 degrees C. This finding suggests that rats prefer SO in a cold environment. There was no significant difference in the ratio of calories of SO ingested to that of matter (standard laboratory chow plus SO) ingested between rats exposed to cold and those at 25 +/- 2 degrees C. The high SO intake also affected cold tolerance and metabolite kinetics in the rats. Factors that affected the SO intake of rats exposed to cold are also discussed.

Barrier Free Site Design.

ERIC Educational Resources Information Center

Dee, Richard K., Ed.

The booklet provides information for the design and evaluation of a barrier free outdoor environment for handicapped individuals. Section 1 discusses the scope of the study, defines terms, cites pertinent laws and legislation, describes cost/benefit factors, and surveys population statistics. Section 2 considers recommended design details in the…

Defining quality in radiology.

PubMed

Blackmore, C Craig

2007-04-01

The introduction of pay for performance in medicine represents an opportunity for radiologists to define quality in radiology. Radiology quality can be defined on the basis of the production model that currently drives reimbursement, codifying the role of radiologists as being limited to the production of timely and accurate radiology reports produced in conditions of maximum patient safety and communicated in a timely manner. Alternately, quality in radiology can also encompass the professional role of radiologists as diagnostic imaging specialists responsible for the appropriate use, selection, interpretation, and application of imaging. Although potentially challenging to implement, the professional model for radiology quality is a comprehensive assessment of the ways in which radiologists add value to patient care. This essay is a discussion of the definition of radiology quality and the implications of that definition.

Current Dilemmas in Defining the Boundaries of Disease.

PubMed

Doust, Jenny; Jean Walker, Mary; Rogers, Wendy A

2017-08-01

Boorse's biostatistical theory states that diseases should be defined in ways that reflect disturbances of biological function and that are objective and value free. We use three examples from contemporary medicine that demonstrate the complex issues that arise when defining the boundaries of disease: polycystic ovary syndrome, chronic kidney disease, and myocardial infarction. We argue that the biostatistical theory fails to provide sufficient guidance on where the boundaries of disease should be drawn, contains ambiguities relating to choice of reference class, and is out of step with medical processes for identifying disease boundaries. Although proponents of the biostatistical theory might regard these practical issues as irrelevant to the aim of providing a theoretical account of disease, we take them to indicate the need for a theoretical account that is adequate for current needs-including limiting new forms of medicalization that are driven by the identification of disease based on dysfunction. Our processes for determining the boundaries for disease need to recognize that there is no value-free method for making these decisions. © The Author 2017. Published by Oxford University Press, on behalf of the Journal of Medicine and Philosophy Inc. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

Ultrasound assisted enzyme catalyzed hydrolysis of waste cooking oil under solvent free condition.

PubMed

Waghmare, Govind V; Rathod, Virendra K

2016-09-01

The present work demonstrates the hydrolysis of waste cooking oil (WCO) under solvent free condition using commercial available immobilized lipase (Novozyme 435) under the influence of ultrasound irradiation. The process parameters were optimized using a sequence of experimental protocol to evaluate the effects of temperature, molar ratios of substrates, enzyme loading, duty cycle and ultrasound intensity. It has been observed that ultrasound-assisted lipase-catalyzed hydrolysis of WCO would be a promising alternative for conventional methods. A maximum conversion of 75.19% was obtained at mild operating parameters: molar ratio of oil to water (buffer pH 7) 3:1, catalyst loading of 1.25% (w/w), lower ultrasound power 100W (ultrasound intensity - 7356.68Wm(-2)), duty cycle 50% and temperature (50°C) in a relatively short reaction time (2h). The activation energy and thermodynamic study shows that the hydrolysis reaction is more feasible when ultrasound is combined with mechanical agitation as compared with the ultrasound alone and simple conventional stirring technique. Application of ultrasound considerably reduced the reaction time as compared to conventional reaction. The successive use of the catalyst for repetitive cycles under the optimum experimental conditions resulted in a loss of enzymatic activity and also minimized the product conversion. Copyright © 2016. Published by Elsevier B.V.

48 CFR 252.225-7013 - Duty-free entry.

Code of Federal Regulations, 2010 CFR

2010-10-01

... the Buy American Act-Free Trade Agreements-Balance of Payments Program clause of this contract; or (iii) Canadian end product as defined in Alternate I of the Buy American Act-Free Trade Agreements... product have the meanings given in the Trade Agreements clause, the Buy American Act and Balance of...

Computational study of small molecule binding for both tethered and free conditions

PubMed Central

2010-01-01

Using a calix[4]arene-benzene complex as a test system we compare the potential of mean force for when the calix[4]arene is tethered versus free. When the complex is in vacuum our results show that the difference between tethered and free is primarily due to the entropic contribution to the potential of mean force resulting in a significant binding free energy difference of 6.6 kJ/mol. By contrast, when the complex is in water our results suggest that there is no appreciable difference between tethered and free. This study elucidates the roles of entropy and enthalpy for this small molecule system and emphasizes the point that tethering the receptor has the potential to dramatically impact the binding properties. These findings should be taken into consideration when using calixarene molecules in nanosensor design. PMID:20369865

Ego depletion interferes with rule-defined category learning but not non-rule-defined category learning.

PubMed

Minda, John P; Rabi, Rahel

2015-01-01

Considerable research on category learning has suggested that many cognitive and environmental factors can have a differential effect on the learning of rule-defined (RD) categories as opposed to the learning of non-rule-defined (NRD) categories. Prior research has also suggested that ego depletion can temporarily reduce the capacity for executive functioning and cognitive flexibility. The present study examined whether temporarily reducing participants' executive functioning via a resource depletion manipulation would differentially impact RD and NRD category learning. Participants were either asked to write a story with no restrictions (the control condition), or without using two common letters (the ego depletion condition). Participants were then asked to learn either a set of RD categories or a set of NRD categories. Resource depleted participants performed more poorly than controls on the RD task, but did not differ from controls on the NRD task, suggesting that self regulatory resources are required for successful RD category learning. These results lend support to multiple systems theories and clarify the role of self-regulatory resources within this theory.

Ego depletion interferes with rule-defined category learning but not non-rule-defined category learning

PubMed Central

Minda, John P.; Rabi, Rahel

2015-01-01

Considerable research on category learning has suggested that many cognitive and environmental factors can have a differential effect on the learning of rule-defined (RD) categories as opposed to the learning of non-rule-defined (NRD) categories. Prior research has also suggested that ego depletion can temporarily reduce the capacity for executive functioning and cognitive flexibility. The present study examined whether temporarily reducing participants’ executive functioning via a resource depletion manipulation would differentially impact RD and NRD category learning. Participants were either asked to write a story with no restrictions (the control condition), or without using two common letters (the ego depletion condition). Participants were then asked to learn either a set of RD categories or a set of NRD categories. Resource depleted participants performed more poorly than controls on the RD task, but did not differ from controls on the NRD task, suggesting that self regulatory resources are required for successful RD category learning. These results lend support to multiple systems theories and clarify the role of self-regulatory resources within this theory. PMID:25688220

Scalable topographies to support proliferation and Oct4 expression by human induced pluripotent stem cells.

PubMed

Reimer, Andreas; Vasilevich, Aliaksei; Hulshof, Frits; Viswanathan, Priyalakshmi; van Blitterswijk, Clemens A; de Boer, Jan; Watt, Fiona M

2016-01-13

It is well established that topographical features modulate cell behaviour, including cell morphology, proliferation and differentiation. To define the effects of topography on human induced pluripotent stem cells (iPSC), we plated cells on a topographical library containing over 1000 different features in medium lacking animal products (xeno-free). Using high content imaging, we determined the effect of each topography on cell proliferation and expression of the pluripotency marker Oct4 24 h after seeding. Features that maintained Oct4 expression also supported proliferation and cell-cell adhesion at 24 h, and by 4 days colonies of Oct4-positive, Sox2-positive cells had formed. Computational analysis revealed that small feature size was the most important determinant of pluripotency, followed by high wave number and high feature density. Using this information we correctly predicted whether any given topography within our library would support the pluripotent state at 24 h. This approach not only facilitates the design of substrates for optimal human iPSC expansion, but also, potentially, identification of topographies with other desirable characteristics, such as promoting differentiation.

Conditional Versus Unconditional Procedures for Sample-Free Item Analysis

ERIC Educational Resources Information Center

Wright, Benjamin D.; Douglas, Graham A.

1977-01-01

Procedures for the Rasch model, sample free item calibration are reviewed and compared for accuracy. The theoretically ideal procedure is shown to have practical limitations. Two alternatives to the ideal are presented and discussed. A correction for bias in the most widely used alternative is presented. (Author/JKS)

Defined Conditions for the Isolation and Expansion of Basal Prostate Progenitor Cells of Mouse and Human Origin

PubMed Central

Höfner, Thomas; Eisen, Christian; Klein, Corinna; Rigo-Watermeier, Teresa; Goeppinger, Stephan M.; Jauch, Anna; Schoell, Brigitte; Vogel, Vanessa; Noll, Elisa; Weichert, Wilko; Baccelli, Irène; Schillert, Anja; Wagner, Steve; Pahernik, Sascha; Sprick, Martin R.; Trumpp, Andreas

2015-01-01

Summary Methods to isolate and culture primary prostate epithelial stem/progenitor cells (PESCs) have proven difficult and ineffective. Here, we present a method to grow and expand both murine and human basal PESCs long term in serum- and feeder-free conditions. The method enriches for adherent mouse basal PESCs with a Lin−SCA-1+CD49f+TROP2high phenotype. Progesterone and sodium selenite are additionally required for the growth of human Lin−CD49f+TROP2high PESCs. The gene-expression profiles of expanded basal PESCs show similarities to ESCs, and NF-kB function is critical for epithelial differentiation of sphere-cultured PESCs. When transplanted in combination with urogenital sinus mesenchyme, expanded mouse and human PESCs generate ectopic prostatic tubules, demonstrating their stem cell activity in vivo. This novel method will facilitate the molecular, genomic, and functional characterization of normal and pathologic prostate glands of mouse and human origin. PMID:25702639

Evaluation of the Omron HJ-720ITC pedometer under free-living conditions.

PubMed

Silcott, Nathan A; Bassett, David R; Thompson, Dixie L; Fitzhugh, Eugene C; Steeves, Jeremy A

2011-09-01

The primary purpose of this study was to examine the accuracy of the Omron HJ-720ITC piezoelectric pedometer in a free-living environment during a 24-h period. A secondary purpose was to compare the accuracy the Omron to that of a spring-levered pedometer. A total of 62 adult volunteers were classified according to body mass index (BMI) category: normal weight (n = 19), overweight (n = 23), and obese (n = 20). Subjects wore five devices during a 24-h period except when bathing or sleeping. The criterion pedometer (StepWatch-3) was worn on the lateral side of the right ankle. Omron HJ-720ITC pedometers were worn in three locations: (a) on the belt, (b) in the pants pocket, and (c) on a lanyard around the neck. A comparison pedometer (Yamax SW-200) was worn on the belt. The Omron HJ-720ITC underestimated the steps per day in all three BMI categories compared with the criterion measure (P < 0.05). The Omron was most accurate in the pocket location, recording 68%, 70%, and 65% of steps in the normal, overweight, and obese BMI categories, respectively. In the obese group, the Omron was more accurate than the Yamax pedometer, but in the normal and overweight groups, it was less accurate than the Yamax. Results from our study show that the Omron significantly underestimates steps per day under free-living conditions. This is due, in part, to the presence of a 4-s step filter that contributes to an underestimation of steps accumulated during intermittent activities. In the pocket, the Omron accuracy was similar for lean, overweight, and obese individuals. In contrast, the accuracy of the Yamax pedometer declined with increasing BMI categories.

A smartphone-driven methodology for estimating physical activities and energy expenditure in free living conditions.

PubMed

Guidoux, Romain; Duclos, Martine; Fleury, Gérard; Lacomme, Philippe; Lamaudière, Nicolas; Manenq, Pierre-Henri; Paris, Ludivine; Ren, Libo; Rousset, Sylvie

2014-12-01

This paper introduces a function dedicated to the estimation of total energy expenditure (TEE) of daily activities based on data from accelerometers integrated into smartphones. The use of mass-market sensors such as accelerometers offers a promising solution for the general public due to the growing smartphone market over the last decade. The TEE estimation function quality was evaluated using data from intensive numerical experiments based, first, on 12 volunteers equipped with a smartphone and two research sensors (Armband and Actiheart) in controlled conditions (CC) and, then, on 30 other volunteers in free-living conditions (FLC). The TEE given by these two sensors in both conditions and estimated from the metabolic equivalent tasks (MET) in CC served as references during the creation and evaluation of the function. The TEE mean gap in absolute value between the function and the three references was 7.0%, 16.4% and 2.7% in CC, and 17.0% and 23.7% according to Armband and Actiheart, respectively, in FLC. This is the first step in the definition of a new feedback mechanism that promotes self-management and daily-efficiency evaluation of physical activity as part of an information system dedicated to the prevention of chronic diseases. Copyright © 2014 Elsevier Inc. All rights reserved.

17 CFR 230.433 - Conditions to permissible post-filing free writing prospectuses.

Code of Federal Regulations, 2014 CFR

2014-04-01

... or transcripts contain all the information provided to the media. (3) For purposes of this paragraph... include information the substance of which is not included in the registration statement. Such a free... format) of any free writing prospectus (including any published article, publication, or advertisement...

17 CFR 230.433 - Conditions to permissible post-filing free writing prospectuses.

Code of Federal Regulations, 2010 CFR

2010-04-01

... or transcripts contain all the information provided to the media. (3) For purposes of this paragraph... include information the substance of which is not included in the registration statement. Such a free... format) of any free writing prospectus (including any published article, publication, or advertisement...

17 CFR 230.433 - Conditions to permissible post-filing free writing prospectuses.

Code of Federal Regulations, 2013 CFR

2013-04-01

... or transcripts contain all the information provided to the media. (3) For purposes of this paragraph... include information the substance of which is not included in the registration statement. Such a free... format) of any free writing prospectus (including any published article, publication, or advertisement...

17 CFR 230.433 - Conditions to permissible post-filing free writing prospectuses.

Code of Federal Regulations, 2012 CFR

2012-04-01

... or transcripts contain all the information provided to the media. (3) For purposes of this paragraph... include information the substance of which is not included in the registration statement. Such a free... format) of any free writing prospectus (including any published article, publication, or advertisement...

17 CFR 230.433 - Conditions to permissible post-filing free writing prospectuses.

Code of Federal Regulations, 2011 CFR

2011-04-01

... or transcripts contain all the information provided to the media. (3) For purposes of this paragraph... include information the substance of which is not included in the registration statement. Such a free... format) of any free writing prospectus (including any published article, publication, or advertisement...

Increased cell-free DNA concentrations in patients with obstructive sleep apnea.

PubMed

Shin, Chol; Kim, Jin K; Kim, Je H; Jung, Ki H; Cho, Kyung J; Lee, Chang K; Lee, Seung G

2008-12-01

Blood concentrations of cell-free DNA, which is considered to be released during apoptosis, are elevated under some pathological conditions such as cardiovascular disease and cancer. The association between obstructive sleep apnea (OSA) and cell-free DNA concentrations has not been reported so far. The purpose of the present study was to examine the association between OSA and plasma DNA concentrations. A case-control study was conducted using a total of 164 men aged 39-67 years, who were free of coronary heart disease and cancer. Laboratory-based overnight polysomnography was performed for all participants. On the basis of polysomnography, patients with an apnea-hypopnea index (AHI) = 5-30 events/h were defined as having mild-moderate OSA (n = 33) and those with >30 events/h were defined as having severe OSA (n = 49). All 82 controls had AHI < 5 events/h. Plasma DNA concentrations from all participants were analyzed for the beta-globin gene using fluorescence-based real-time polymerase chain reaction. Patients with severe OSA had significantly higher plasma DNA concentrations than persons with mild-moderate OSA and those without OSA (P < 0.05). AHI was significantly associated with body mass index (P < 0.001), hypertension (P < 0.001), and plasma DNA concentration (P < 0.05). After taking into account hypertension and other potential risk factors, persons with high plasma DNA concentrations (>8 microg/L) had approximately fourfold higher odds of OSA than those with low DNA levels. Further data are warranted to confirm the association for men and to evaluate the association for women.

A cell-free framework for rapid biosynthetic pathway prototyping and enzyme discovery.

PubMed

Karim, Ashty S; Jewett, Michael C

2016-07-01

Speeding up design-build-test (DBT) cycles is a fundamental challenge facing biochemical engineering. To address this challenge, we report a new cell-free protein synthesis driven metabolic engineering (CFPS-ME) framework for rapid biosynthetic pathway prototyping. In our framework, cell-free cocktails for synthesizing target small molecules are assembled in a mix-and-match fashion from crude cell lysates either containing selectively enriched pathway enzymes from heterologous overexpression or directly producing pathway enzymes in lysates by CFPS. As a model, we apply our approach to n-butanol biosynthesis showing that Escherichia coli lysates support a highly active 17-step CoA-dependent n-butanol pathway in vitro. The elevated degree of flexibility in the cell-free environment allows us to manipulate physiochemical conditions, access enzymatic nodes, discover new enzymes, and prototype enzyme sets with linear DNA templates to study pathway performance. We anticipate that CFPS-ME will facilitate efforts to define, manipulate, and understand metabolic pathways for accelerated DBT cycles without the need to reengineer organisms. Copyright © 2016 International Metabolic Engineering Society. Published by Elsevier Inc. All rights reserved.

Visual space under free viewing conditions.

PubMed

Doumen, Michelle J A; Kappers, Astrid M L; Koenderink, Jan J

2005-10-01

Most research on visual space has been done under restricted viewing conditions and in reduced environments. In our experiments, observers performed an exocentric pointing task, a collinearity task, and a parallelity task in a entirely visible room. We varied the relative distances between the objects and the observer and the separation angle between the two objects. We were able to compare our data directly with data from experiments in an environment with less monocular depth information present. We expected that in a richer environment and under less restrictive viewing conditions, the settings would deviate less from the veridical settings. However, large systematic deviations from veridical settings were found for all three tasks. The structure of these deviations was task dependent, and the structure and the deviations themselves were comparable to those obtained under more restricted circumstances. Thus, the additional information was not used effectively by the observers.

Validity of the SenseWear armband step count measure during controlled and free-living conditions.

PubMed

Lee, Joey Allen; Laurson, Kelly Rian

2015-06-01

Advances in technology continue to provide numerous options for physical activity assessment. These advances necessitate evaluation of the validity of newly developed activity monitors being used in clinical and research settings. The purpose of this study was to validate the SenseWear Pro3 Armband (SWA) step counts during treadmill walking and free-living conditions. Study 1 observed 39 individuals (17 males, 22 females) wearing an SWA and a Yamax Digiwalker SW-701 pedometer (DIGI) during treadmill walking, utilizing manually counted steps as the criterion. Study 2 compared free-living step count data from 35 participants (17 males, 18 females) wearing the SWA and DIGI (comparison) for 3 consecutive days. During Study 1, the SWA underestimated steps by 16.0%, 10.7%, 5.6%, 6.1%, and 6.5% at speeds of 54 m/min, 67 m/min, 80 m/min, 94 m/min, and 107 m/min, respectively, compared to manually counted steps. During Study 2, the intraclass correlation (ICC) coefficient of mean steps/d between the SWA and DIGI was strong (r = 0.98, p  < 0.001). Unlike Study 1, the SWA overestimated step counts during the 3-day wear period by an average of 1028 steps/d (or +11.3%) compared to the DIGI. When analyzed individually, the SWA consistently overestimated step counts for each day ( p  < 0.05). The SWA underestimates steps during treadmill walking and appears to overestimate steps during free-living compared to the DIGI pedometer. Caution is warranted when using the SWA to count steps. Modifications are needed to enhance step counting accuracy.

Defined PEG smears as an alternative approach to enhance the search for crystallization conditions and crystal-quality improvement in reduced screens

PubMed Central

Chaikuad, Apirat; Knapp, Stefan; von Delft, Frank

2015-01-01

The quest for an optimal limited set of effective crystallization conditions remains a challenge in macromolecular crystallography, an issue that is complicated by the large number of chemicals which have been deemed to be suitable for promoting crystal growth. The lack of rational approaches towards the selection of successful chemical space and representative combinations has led to significant overlapping conditions, which are currently present in a multitude of commercially available crystallization screens. Here, an alternative approach to the sampling of widely used PEG precipitants is suggested through the use of PEG smears, which are mixtures of different PEGs with a requirement of either neutral or cooperatively positive effects of each component on crystal growth. Four newly defined smears were classified by molecular-weight groups and enabled the preservation of specific properties related to different polymer sizes. These smears not only allowed a wide coverage of properties of these polymers, but also reduced PEG variables, enabling greater sampling of other parameters such as buffers and additives. The efficiency of the smear-based screens was evaluated on more than 220 diverse recombinant human proteins, which overall revealed a good initial crystallization success rate of nearly 50%. In addition, in several cases successful crystallizations were only obtained using PEG smears, while various commercial screens failed to yield crystals. The defined smears therefore offer an alternative approach towards PEG sampling, which will benefit the design of crystallization screens sampling a wide chemical space of this key precipitant. PMID:26249344

Defined PEG smears as an alternative approach to enhance the search for crystallization conditions and crystal-quality improvement in reduced screens.

PubMed

Chaikuad, Apirat; Knapp, Stefan; von Delft, Frank

2015-08-01

The quest for an optimal limited set of effective crystallization conditions remains a challenge in macromolecular crystallography, an issue that is complicated by the large number of chemicals which have been deemed to be suitable for promoting crystal growth. The lack of rational approaches towards the selection of successful chemical space and representative combinations has led to significant overlapping conditions, which are currently present in a multitude of commercially available crystallization screens. Here, an alternative approach to the sampling of widely used PEG precipitants is suggested through the use of PEG smears, which are mixtures of different PEGs with a requirement of either neutral or cooperatively positive effects of each component on crystal growth. Four newly defined smears were classified by molecular-weight groups and enabled the preservation of specific properties related to different polymer sizes. These smears not only allowed a wide coverage of properties of these polymers, but also reduced PEG variables, enabling greater sampling of other parameters such as buffers and additives. The efficiency of the smear-based screens was evaluated on more than 220 diverse recombinant human proteins, which overall revealed a good initial crystallization success rate of nearly 50%. In addition, in several cases successful crystallizations were only obtained using PEG smears, while various commercial screens failed to yield crystals. The defined smears therefore offer an alternative approach towards PEG sampling, which will benefit the design of crystallization screens sampling a wide chemical space of this key precipitant.

Sleep-wake cycle of an unrestrained isolated chimpanzee under entrained and free running conditions.

NASA Technical Reports Server (NTRS)

Mcnew, J. J.; Burson, R. C.; Hoshizaki, T.; Adey, W. R.

1972-01-01

Biorhythmic patterns of EEG activity - the sleep-wake cycle and the sleep cycle - were investigated in an unrestrained chimpanzee subjected to 30 days of isolation in a 4-ft cubical cage placed in a high performance sound isolation chamber. The animal received 10 days of 12 hours of light and 12 hours of dark, then 10 days of continuous light, followed by 10 more days of 12 hours of light and 12 hours of dark. The circadian sleep-wake rhythm and the wake and sleep phases of this rhythm during entrained and free running conditions were analyzed in terms of duration. The awake and nonREM sleep and REM sleep stages were also analyzed. In addition, the mean duration of the sleep cycle of the sleep phase was computed.

Free Vibration Study of Anti-Symmetric Angle-Ply Laminated Plates under Clamped Boundary Conditions

NASA Astrophysics Data System (ADS)

Viswanathan, K. K.; Karthik, K.; Sanyasiraju, Y. V. S. S.; Aziz, Z. A.

2016-11-01

Two type of numerical approach namely, Radial Basis Function and Spline approximation, used to analyse the free vibration of anti-symmetric angle-ply laminated plates under clamped boundary conditions. The equations of motion are derived using YNS theory under first order shear deformation. By assuming the solution in separable form, coupled differential equations obtained in term of mid-plane displacement and rotational functions. The coupled differential is then approximated using Spline function and radial basis function to obtain the generalize eigenvalue problem and parametric studies are made to investigate the effect of aspect ratio, length-to-thickness ratio, number of layers, fibre orientation and material properties with respect to the frequency parameter. Some results are compared with the existing literature and other new results are given in tables and graphs.

CATALYST-FREE REACTIONS UNDER SOLVENT-FEE CONDITIONS: MICROWAVE-ASSISTED SYNTHESIS OF HETEROCYCLIC HYDRAZONES BELOW THE MELTING POINT OF NEAT REACTANTS: JOURNAL ARTICLE

EPA Science Inventory

NRMRL-CIN-1437 Jeselnik, M., Varma*, R.S., Polanc, S., and Kocevar, M. Catalyst-free Reactions under Solvent-fee Conditions: Microwave-assisted Synthesis of Heterocyclic Hydrazones below the Melting Point of Neat Reactants. Published in: Chemical Communications 18:1716-1717 (200...

Cinematographic analysis of bovine embryo development in serum-free oviduct-conditioned medium.

PubMed

Grisart, B; Massip, A; Dessy, F

1994-07-01

Development of bovine embryos produced in vitro from the one-cell to the blastocyst stage in serum-free oviduct-conditioned medium was investigated for 8 days consecutively by time-lapse cinematography. Three movies were analysed (130 embryos). The following observations were made. (1) Development under cine-recording conditions was similar to that in a classical incubator. (2) The highest proportion of embryos at the two-cell, three-four-cell, five-eight-cell, 9-16-cell, morula and blastocyst stages were recorded at 34, 46, 61, 115, 149 and 192 h after insemination, respectively. Cleavage asynchrony between blastomeres within individual embryos started at the two-cell stage. (3) The duration of the first three cell cycles was 35 h, 14 h and 11-62 h, respectively. (4) Detailed analysis of 13 embryos revealed that developmental arrest ('Lag-phase') occurred at the four-cell (1 of 13), five-cell (2 of 13), six-cell (3 of 13), seven-cell (3 of 13) or eight-cell stage (4 of 13); this phase lasted about 59 h. Embryos arrested at the eight-cell stage developed into morula-blastocysts (3 of 4) at a higher rate than did those arrested at earlier stages (2 of 9). (5) The faster the embryos cleaved into early stages (two-cell, three-four-cell and five-eight-cell), the higher the probability that they developed into morula-blastocyst: 70% of the embryos reaching the two-cell stage before 30-31 h after insemination developed into morula-blastocyst.(ABSTRACT TRUNCATED AT 250 WORDS)

The complexity of treating wasting in ambulatory rehabilitation: Is it starvation, sarcopenia, cachexia or a combination of these conditions?

PubMed

Yaxley, Alison; Miller, Michelle D; Fraser, Robert J; Cobiac, Lynne; Crotty, Maria

2012-01-01

Nutritional status is often impaired in ambulatory rehabilitation patients. Wasting conditions can be classified as starvation, sarcopenia or cachexia but differences between these are not well defined, and misdiagnosis may lead to inappropriate intervention. A secondary analysis of data from 187 ambulatory rehabilitation patients aged >=60 years aimed to identify patients with one or more wasting condition, and investigate the impact on common rehabilitation outcomes. Starvation was defined by fat-free mass index and the Council on Nutrition Appetite Questionnaire score; sarcopenia by fat-free mass index and quadriceps strength; and cachexia by fat-free mass index and serum C-reactive protein. Selected rehabilitation outcomes were compared for those who were, and those who were not, identified as having one or more wasting condition. Of those identified with starvation (n=30), all were also identified as sarcopenic and 20 as cachectic; of those identified as sarcopenic (n=75), 30 had starvation and 37 were cachectic; and of those identified as cachectic (n=37), 20 had starvation and all were sarcopenic. Twenty participants were identified as having all three conditions. Those with starvation had higher level of depression (p=0.003), lower self-rated health (p=0.032), and lower levels of physical function (motor p=0.006; process p=0.004) than those with no evidence of a wasting condition. Those who had sarcopenia had lower physical function (motor p=0.012; process p=0.003) as did those with cachexia (motor p=0.025; process p=0.042). Results suggest problems in operationalising definitions in an ambulatory clinical setting. The overlap identified in this analysis suggests that up to 40% (75/187) of patients could be misidentified and prescribed inappropriate nutritional support.

Rapid Induction of Cerebral Organoids From Human Induced Pluripotent Stem Cells Using a Chemically Defined Hydrogel and Defined Cell Culture Medium.

PubMed

Lindborg, Beth A; Brekke, John H; Vegoe, Amanda L; Ulrich, Connor B; Haider, Kerri T; Subramaniam, Sandhya; Venhuizen, Scott L; Eide, Cindy R; Orchard, Paul J; Chen, Weili; Wang, Qi; Pelaez, Francisco; Scott, Carolyn M; Kokkoli, Efrosini; Keirstead, Susan A; Dutton, James R; Tolar, Jakub; O'Brien, Timothy D

2016-07-01

Tissue organoids are a promising technology that may accelerate development of the societal and NIH mandate for precision medicine. Here we describe a robust and simple method for generating cerebral organoids (cOrgs) from human pluripotent stem cells by using a chemically defined hydrogel material and chemically defined culture medium. By using no additional neural induction components, cOrgs appeared on the hydrogel surface within 10-14 days, and under static culture conditions, they attained sizes up to 3 mm in greatest dimension by day 28. Histologically, the organoids showed neural rosette and neural tube-like structures and evidence of early corticogenesis. Immunostaining and quantitative reverse-transcription polymerase chain reaction demonstrated protein and gene expression representative of forebrain, midbrain, and hindbrain development. Physiologic studies showed responses to glutamate and depolarization in many cells, consistent with neural behavior. The method of cerebral organoid generation described here facilitates access to this technology, enables scalable applications, and provides a potential pathway to translational applications where defined components are desirable. Tissue organoids are a promising technology with many potential applications, such as pharmaceutical screens and development of in vitro disease models, particularly for human polygenic conditions where animal models are insufficient. This work describes a robust and simple method for generating cerebral organoids from human induced pluripotent stem cells by using a chemically defined hydrogel material and chemically defined culture medium. This method, by virtue of its simplicity and use of defined materials, greatly facilitates access to cerebral organoid technology, enables scalable applications, and provides a potential pathway to translational applications where defined components are desirable. ©AlphaMed Press.

Ultrasensitive environmental assessment of xeno-estrogens in water samples using label-free graphene immunosensors.

PubMed

Barton, Huw; Berbel-Filho, Waldir M; Consuegra, Sofia; Francis, Lewis; Tizaoui, Chedly; Conlan, R Steven; Teixeira, Sofia Rodrigues

2018-05-01

There is a growing interest in the possible environmental health impact posed by endocrine-disrupting chemicals (EDCs). A challenge to the field of endocrine disruption is that these substances are diverse and may not appear to share any structural similarity other than usually being low molecular mass (<1000 Da) compounds. Here we demonstrate the effectiveness of sensor device for the detection of low molecular weight, poorly water soluble, estrogenic compounds E1, E2 and EE2, fabricated by electropolymerization over graphene screen printed electrode (SPE). The PANI/Gr-SPE-devices displayed linear responses to estrogenic substances, in EIS assays, from 0.0975 ng/L to 200 ng/L in water samples, with a detection limit of 0.043 pg/L for E1, 0.19 ng/L for E2 and 0.070 pg/L for EE2 which is lower than other current biosensing techniques. This portable, disposable immunosensor offers a solution for immediate measurement at sample collection sites, due to its excellent sensitivity and selectivity when testing water samples obtained directly from rivers and waste water treatment facilities. The simple screen printing production method will enable the low cost, high volume production required for this type of environmental analysis. Copyright © 2018 Elsevier Inc. All rights reserved.

Influence of chemistry, interfacial width, and non-isothermal conditions on spatially heterogeneous activated relaxation and elasticity in glass-forming free standing films

DOE Office of Scientific and Technical Information (OSTI.GOV)

Mirigian, Stephen; Schweizer, Kenneth S.

Here, we employ the Elastically Collective Nonlinear Langevin Equation (ECNLE) theory of activated relaxation to study several questions in free standing thin films of glass-forming molecular and polymer liquids. The influence of non-universal chemical aspects on dynamical confinement effects is found to be relatively weak, but with the caveat that for the systems examined, the bulk ECNLE polymer theory does not predict widely varying fragilities. Allowing the film model to have a realistic vapor interfacial width significantly enhances the reduction of the film-averaged glass transition temperature, T g, in a manner that depends on whether a dynamic or pseudo-thermodynamic averagingmore » of the spatial mobility gradient is adopted. The nature of film thickness effects on the spatial profiles of the alpha relaxation time and elastic modulus is studied under non-isothermal conditions and contrasted with the corresponding isothermal behavior. Modest differences are found if a film-thickness dependent T g is defined in a dynamical manner. But, adopting a pseudo-thermodynamic measure of T g leads to a qualitatively new form of the alpha relaxation time gradient where highly mobile layers near the film surface coexist with strongly vitrified regions in the film interior. Consequently, the film-averaged shear modulus can increase with decreasing film thickness, despite the T g reduction and presence of a mobile surface layer. Such a behavior stands in qualitative contrast to the predicted mechanical softening under isothermal conditions. Spatial gradients of the elastic modulus are studied as a function of temperature, film thickness, probing frequency, and experimental protocol, and a rich behavior is found.« less

Influence of chemistry, interfacial width, and non-isothermal conditions on spatially heterogeneous activated relaxation and elasticity in glass-forming free standing films

DOE PAGES

Mirigian, Stephen; Schweizer, Kenneth S.

2017-02-02

Here, we employ the Elastically Collective Nonlinear Langevin Equation (ECNLE) theory of activated relaxation to study several questions in free standing thin films of glass-forming molecular and polymer liquids. The influence of non-universal chemical aspects on dynamical confinement effects is found to be relatively weak, but with the caveat that for the systems examined, the bulk ECNLE polymer theory does not predict widely varying fragilities. Allowing the film model to have a realistic vapor interfacial width significantly enhances the reduction of the film-averaged glass transition temperature, T g, in a manner that depends on whether a dynamic or pseudo-thermodynamic averagingmore » of the spatial mobility gradient is adopted. The nature of film thickness effects on the spatial profiles of the alpha relaxation time and elastic modulus is studied under non-isothermal conditions and contrasted with the corresponding isothermal behavior. Modest differences are found if a film-thickness dependent T g is defined in a dynamical manner. But, adopting a pseudo-thermodynamic measure of T g leads to a qualitatively new form of the alpha relaxation time gradient where highly mobile layers near the film surface coexist with strongly vitrified regions in the film interior. Consequently, the film-averaged shear modulus can increase with decreasing film thickness, despite the T g reduction and presence of a mobile surface layer. Such a behavior stands in qualitative contrast to the predicted mechanical softening under isothermal conditions. Spatial gradients of the elastic modulus are studied as a function of temperature, film thickness, probing frequency, and experimental protocol, and a rich behavior is found.« less

Simulating recurrent event data with hazard functions defined on a total time scale.

PubMed

Jahn-Eimermacher, Antje; Ingel, Katharina; Ozga, Ann-Kathrin; Preussler, Stella; Binder, Harald

2015-03-08

In medical studies with recurrent event data a total time scale perspective is often needed to adequately reflect disease mechanisms. This means that the hazard process is defined on the time since some starting point, e.g. the beginning of some disease, in contrast to a gap time scale where the hazard process restarts after each event. While techniques such as the Andersen-Gill model have been developed for analyzing data from a total time perspective, techniques for the simulation of such data, e.g. for sample size planning, have not been investigated so far. We have derived a simulation algorithm covering the Andersen-Gill model that can be used for sample size planning in clinical trials as well as the investigation of modeling techniques. Specifically, we allow for fixed and/or random covariates and an arbitrary hazard function defined on a total time scale. Furthermore we take into account that individuals may be temporarily insusceptible to a recurrent incidence of the event. The methods are based on conditional distributions of the inter-event times conditional on the total time of the preceeding event or study start. Closed form solutions are provided for common distributions. The derived methods have been implemented in a readily accessible R script. The proposed techniques are illustrated by planning the sample size for a clinical trial with complex recurrent event data. The required sample size is shown to be affected not only by censoring and intra-patient correlation, but also by the presence of risk-free intervals. This demonstrates the need for a simulation algorithm that particularly allows for complex study designs where no analytical sample size formulas might exist. The derived simulation algorithm is seen to be useful for the simulation of recurrent event data that follow an Andersen-Gill model. Next to the use of a total time scale, it allows for intra-patient correlation and risk-free intervals as are often observed in clinical trial data

Evidence Gaps in the Use of Spinal Cord Stimulation for Treating Chronic Spine Conditions.

PubMed

Provenzano, David A; Amirdelfan, Kasra; Kapural, Leonardo; Sitzman, B Todd

2017-07-15

A review of literature. The aim of this study was to define and explore the current evidence gaps in the use of spinal cord stimulation (SCS) for treating chronic spine conditions. Although over the last 40 years SCS therapy has undergone significant technological advancements, evidence gaps still exist. A literature review was conducted to define current evidence gaps for the use of SCS. Areas of focus included 1) treatment of cervical spine conditions, 2) treatment of lumbar spine conditions, 3) technological advancement and device selection, 4) appropriate patient selection, 5) the ability to curb pharmacological treatment, and 6) methods to prolong efficacy over time. New SCS strategies using advanced waveforms are explored. The efficacy, safety, and cost-effectiveness of traditional SCS for chronic pain conditions are well-established. Evidence gaps do exist. Recently, advancement in waveforms and programming parameters have allowed for paresthesia-reduced/free stimulation that in specific clinical areas may improve clinical outcomes. New waveforms such as 10-kHz high-frequency have resulted in an improvement in back coverage. To date, clinical efficacy data are more prevalent for the treatment of painful conditions originating from the lumbar spine in comparison to the cervical spine. Evidence gaps still exist that require appropriate study designs with long-term follow-up to better define and improve the use of this therapy for the treatment of chronic spine pain in both the cervical and lumbar regions. N/A.

Bond strengths of different orthodontic adhesives after enamel conditioning with the same self-etching primer.

PubMed

Scougall-Vilchis, Rogelio J; Zárate-Díaz, Chrisel; Kusakabe, Shusuke; Yamamoto, Kohji

2010-05-01

To determine the shear bond strengths (SBS) of stainless steel brackets bonded with seven light-cured orthodontic adhesives after the enamel was conditioned with the same self-etching primer. A total of 140 extracted human molars were randomly divided into seven groups (N = 20). In all the groups, the enamel was conditioned with Transbond Plus SEP (TPSEP). Stainless steel brackets were bonded with the following orthodontic adhesives: Group I, Transbond XT; Group II, Blūgloo; Group III, BeautyOrtho Bond; Group IV, Enlight; Group V, Light Bond; Group VI, Transbond CC; Group VII, Xeno Ortho. The teeth were stored in distilled water at 37 degrees C for 24 hours and debonded with a universal testing machine. The modified adhesive remnant index (ARI) was also recorded. There were no significant differences in the SBS values among the groups: I (18.0 +/- 7.4 MPa); II (18.3 +/- 5.1 MPa); III (14.8 +/- 4.3 MPa); IV (18.3 +/- 7.0 MPa); V (16.4 +/- 4.3 MPa); VI (20.3 +/- 5.3 MPa); VII (15.9 +/- 6.4 MPa), but significant differences in ARI were found. The seven orthodontic adhesives evaluated in this study can be successfully used for bonding stainless steel brackets when the enamel is conditioned with TPSEP, however, the differences among some groups might influence the clinical bond strengths. In addition, the amount of residual adhesive remaining on the teeth after debonding differed among the adhesives. Further studies are required to better understand the differences in SBS and ARI.

Epidermal growth factor- and hepatocyte growth factor-receptor activity in serum-free cultures of human hepatocytes.

PubMed

Runge, D M; Runge, D; Dorko, K; Pisarov, L A; Leckel, K; Kostrubsky, V E; Thomas, D; Strom, S C; Michalopoulos, G K

1999-02-01

Serum-free primary cultures of hepatocytes are a useful tool to study factors triggering hepatocyte proliferation and regeneration. We have developed a chemically defined serum-free system that allows human hepatocyte proliferation in the presence of epidermal growth factor and hepatocyte growth factor. DNA synthesis and accumulation were determined by [3H]thymidine incorporation and fluorometry, respectively. Western blot analyses and co-immunoprecipitations were used to investigate the association of proteins involved in epidermal growth factor and hepatocyte growth factor activation and signaling: epidermal growth factor receptor, hepatocyte growth factor receptor (MET), urokinase-type plasminogen activator and its receptor, and a member of the signal transducer and activator of transcription family, STAT-3. Primary human hepatocytes proliferated under serum-free conditions in a chemically defined medium for up to 12 days. Epidermal growth factor-receptor and MET were present and functional, decreasing over time. MET, urokinase-type plasminogen activator and urokinase-type plasminogen activator receptor co-precipitated to varying degrees during the culture period. STAT-3 co-precipitated with epidermal growth factor-receptor and MET to varying degrees. Proliferation of human hepatocytes can improve by modification of a chemically defined medium originally used for rat hepatocyte cultures. In these long-term cultures of human hepatocytes, hepatocyte growth factor and epidermal growth factor can stimulate growth and differentiation by interacting with their receptors and initiating downstream signaling. This involves complex formation of the receptors with other plasma membrane components for MET (urokinase-type plasminogen activator in context of its receptor) and activation of STAT-3 for both receptors.

Compound cuing in free recall.

PubMed

Lohnas, Lynn J; Kahana, Michael J

2014-01-01

According to the retrieved context theory of episodic memory, the cue for recall of an item is a weighted sum of recently activated cognitive states, including previously recalled and studied items as well as their associations. We show that this theory predicts there should be compound cuing in free recall. Specifically, the temporal contiguity effect should be greater when the 2 most recently recalled items were studied in contiguous list positions. A meta-analysis of published free recall experiments demonstrates evidence for compound cuing in both conditional response probabilities and interresponse times. To help rule out a rehearsal-based account of these compound cuing effects, we conducted an experiment with immediate, delayed, and continual-distractor free recall conditions. Consistent with retrieved context theory but not with a rehearsal-based account, compound cuing was present in all conditions, and was not significantly influenced by the presence of interitem distractors.

Compound cueing in free recall

PubMed Central

Lohnas, Lynn J.; Kahana, Michael J.

2013-01-01

According to the retrieved context theory of episodic memory, the cue for recall of an item is a weighted sum of recently activated cognitive states, including previously recalled and studied items as well as their associations. We show that this theory predicts there should be compound cueing in free recall. Specifically, the temporal contiguity effect should be greater when the two most recently recalled items were studied in contiguous list positions. A meta-analysis of published free recall experiments demonstrates evidence for compound cueing in both conditional response probabilities and inter-response times. To help rule out a rehearsal-based account of these compound cueing effects, we conducted an experiment with immediate, delayed and continual-distractor free recall conditions. Consistent with retrieved context theory but not with a rehearsal-based account, compound cueing was present in all conditions, and was not significantly influenced by the presence of interitem distractors. PMID:23957364

Free Radical Mechanisms in Autoxidation Processes.

ERIC Educational Resources Information Center

Simic, Michael G.

1981-01-01

Discusses the use of steady-state radiation chemistry and pulse radiolysis for the generation of initial free radicals and formation of peroxy radicals in the autoxidation process. Provides information regarding the autoxidation process. Defines autoxidation reactions and antioxidant action. (CS)

Characterization of Canine Adipose-Derived Mesenchymal Stromal/Stem Cells in Serum-Free Medium.

PubMed

Liu, Zhuoming; Screven, Rudell; Boxer, Lynne; Myers, Michael J; Devireddy, Lax R

2018-06-20

In this article, we report on the development of a defined serum-free medium capable of supporting the culture expansion of mesenchymal stromal/stem cells (MSCs) from canine adipose tissue (canine Ad-MSCs). The potential benefits of serum-free media can only be utilized if cells cultured in serum-free media maintain the same functional characteristics as cells cultured in serum-containing media. Therefore, we analyze the characteristics of canine Ad-MSCs cultured in this serum-free medium or in serum-containing medium through evaluation of growth kinetics, clonogenic capacity, senescence, and differentiation capacity. Both, serum-containing medium and our serum-free medium, supported efficient growth and colony formation of canine Ad-MSCs. In addition, canine Ad-MSCs cultured in both media demonstrated similar viability after freeze/thaw, similar cell surface marker expression, and were capable of trilineage differentiation. While canine Ad-MSCs cultured in both media were generally similar, under the conditions of our study, canine Ad-MSCs cultured in serum-free medium demonstrated a shorter lag phase and higher colony-forming capacity, accelerated population doubling, maintained multipotentiality at higher passage numbers, and underwent senescence at higher passage numbers compared to canine Ad-MSCs cultured in conventional serum-containing medium. These results suggest that canine Ad-MSCs cultured in serum-free medium retain the basic characteristics associated with canine Ad-MSCs cultured in serum-containing medium, although some differences in growth kinetics were observed.

BPIC: A novel anti-tumor lead capable of inhibiting inflammation and scavenging free radicals.

PubMed

Li, Shan; Wang, Yuji; Zhao, Ming; Wu, Jianhui; Peng, Shiqi

2015-03-01

Inflammation has a critical role in the tumor progression, free radical damage can worse the status of patients in cancer condition. The anti-cancer agents capable of inhibiting inflammation and scavenging free radicals attract a lot of our interest. Aimed at the discovery of such anti-tumor agent, a novel intercalator, benzyl 1-[4-hydroxy-3-(methoxycarbonyl)-phenyl-9H-pyrido[3,4-b]indole-3-carboxylate (BPIC) was presented. The docking investigation of BPIC and doxorubicin towards the DNA (PDB ID: 1NAB) gave equal score and similar feature. The anti-proliferation assay of 8 cancer cells identified S180 cells had equal sensitivity to BPIC and doxorubicin. The anti-tumor assay defined the efficacy of BPIC been 2 folds higher than that of doxorubicin. At 1μmol/kg of dose BPIC effectively inhibited xylene-induced ear edema and decreased the plasma TNF-α and IL-8 of the mice. BPIC scavenged ∙OH, ∙O2(-) and NO free radicals in a concentration dependent manner and NO free radicals had the highest sensitivity. BPIC could be a novel anti-tumor lead capable of simultaneously inhibiting inflammation and scavenging free radicals. Copyright © 2015 Elsevier Ltd. All rights reserved.

Environmental conditions associated with lesions in introduced free-ranging sheep in Hawai‘i

USGS Publications Warehouse

Powers, Jenny G.; Duncan, Colleen G.; Spraker, Terry R.; Schuler, Bridget A.; Hess, Steven C.; Faford, Jonathan K.J.; Sin, Hans

2014-01-01

Wildlife species which have been translocated between temperate and tropical regions of the world provide unique opportunities to understand how disease processes may be affected by environmental conditions. European mouflon sheep (Ovis gmelini musimon) from the Mediterranean Islands were introduced to the Hawaiian Islands for sport hunting beginning in 1954 and were subsequently hybridized with feral domestic sheep (O. aries), which had been introduced in 1793. Three isolated mouflon populations have become established in the Hawaiian Islands but diseases in these populations have been little studied. The objective of this study was to evaluate and compare gross and histologic lesions in respiratory, renal, and hepatic systems of free-ranging sheep in two isolated volcanic environments on Hawai‘i Island. Tissue and fecal samples were collected in conjunction with population reductions during February 2011. We found gross or histologic evidence of lungworm infection in 44/49 sheep from Mauna Loa which were exposed to gaseous emissions from Kīlauea Volcano. In contrast, only 7/50 sheep from Mauna Kea had lesions consistent with lungworm, but Mauna Kea sheep had significantly more upper respiratory tract inflammation and hyperplasia consistent with chronic antigenic stimulation, possibly associated with exposure to fine airborne particulates during extended drought conditions. We hypothesize that gasses from Kīlauea Volcano contributed to severity of respiratory disease principally associated with chronic lungworm infections at Mauna Loa; however, there were numerous other potentially confounding environmental factors and interactions that merit further investigation.

Particle Creation at a Point Source by Means of Interior-Boundary Conditions

NASA Astrophysics Data System (ADS)

Lampart, Jonas; Schmidt, Julian; Teufel, Stefan; Tumulka, Roderich

2018-06-01

We consider a way of defining quantum Hamiltonians involving particle creation and annihilation based on an interior-boundary condition (IBC) on the wave function, where the wave function is the particle-position representation of a vector in Fock space, and the IBC relates (essentially) the values of the wave function at any two configurations that differ only by the creation of a particle. Here we prove, for a model of particle creation at one or more point sources using the Laplace operator as the free Hamiltonian, that a Hamiltonian can indeed be rigorously defined in this way without the need for any ultraviolet regularization, and that it is self-adjoint. We prove further that introducing an ultraviolet cut-off (thus smearing out particles over a positive radius) and applying a certain known renormalization procedure (taking the limit of removing the cut-off while subtracting a constant that tends to infinity) yields, up to addition of a finite constant, the Hamiltonian defined by the IBC.

75 FR 27927 - Diversification Requirements for Certain Defined Contribution Plans

Federal Register 2010, 2011, 2012, 2013, 2014

2010-05-19

... section 414(l) with respect to any other defined benefit plan or defined contribution plan maintained by... disclosure of the fund's portfolio holdings (for example, Form N-CSR, ``Certified Shareholder Report of... securities, as well as a direct or indirect benefit that is conditioned on investment in employer securities...

Vertical pump with free floating check valve

DOEpatents

Lindsay, Malcolm

1980-01-01

A vertical pump with a bottom discharge having a free floating check valve isposed in the outlet plenum thereof. The free floating check valve comprises a spherical member with a hemispherical cage-like member attached thereto which is capable of allowing forward or reverse flow under appropriate conditions while preventing reverse flow under inappropriate conditions.

CHEMISTRY UNDER 'GREENER' CONDITIONS: SOLVENT-FREE SYNTHESIS USING MICROWAVE IRRADIATION

EPA Science Inventory

Solvent-free approach is emphasized that involves microwave (MW) irradiation of neat reactants (undiluted) catalyzed by the surfaces of less-expensive and recyclable mineral supports such as alumina, silica, clay, or 'doped' surfaces which is applicable to a wide range of cleavag...

Free energy decomposition of protein-protein interactions.

PubMed

Noskov, S Y; Lim, C

2001-08-01

A free energy decomposition scheme has been developed and tested on antibody-antigen and protease-inhibitor binding for which accurate experimental structures were available for both free and bound proteins. Using the x-ray coordinates of the free and bound proteins, the absolute binding free energy was computed assuming additivity of three well-defined, physical processes: desolvation of the x-ray structures, isomerization of the x-ray conformation to a nearby local minimum in the gas-phase, and subsequent noncovalent complex formation in the gas phase. This free energy scheme, together with the Generalized Born model for computing the electrostatic solvation free energy, yielded binding free energies in remarkable agreement with experimental data. Two assumptions commonly used in theoretical treatments; viz., the rigid-binding approximation (which assumes no conformational change upon complexation) and the neglect of vdW interactions, were found to yield large errors in the binding free energy. Protein-protein vdW and electrostatic interactions between complementary surfaces over a relatively large area (1400--1700 A(2)) were found to drive antibody-antigen and protease-inhibitor binding.

Culture of bovine ovarian follicle wall sections maintained the highly estrogenic profile under basal and chemically defined conditions

PubMed Central

Vasconcelos, R.B.; Salles, L.P.; Silva, I. Oliveira e; Gulart, L.V.M.; Souza, D.K.; Torres, F.A.G.; Bocca, A.L.; Silva, A.A.M. Rosa e

2013-01-01

Follicle cultures reproduce in vitro the functional features observed in vivo. In a search for an ideal model, we cultured bovine antral follicle wall sections (FWS) in a serum-free defined medium (DM) known to induce 17β-estradiol (E2) production, and in a nondefined medium (NDM) containing serum. Follicles were sectioned and cultured in NDM or DM for 24 or 48 h. Morphological features were determined by light microscopy. Gene expression of steroidogenic enzymes and follicle-stimulating hormone (FSH) receptor were determined by RT-PCR; progesterone (P4) and E2 concentrations in the media were measured by radioimmunoassay. DM, but not NDM, maintained an FWS morphology in vitro that was similar to fresh tissue. DM also induced an increase in the expression of all steroidogenic enzymes, except FSH receptor, but NDM did not. In both DM and NDM, there was a gradual increase in P4 throughout the culture period; however, P4 concentration was significantly higher in NDM. In both media, E2 concentration was increased at 24 h, followed by a decrease at 48 h. The E2:P4 ratio was higher in DM than in NDM. These results suggest that DM maintains morphological structure, upregulates the expression of steroidogenic enzyme genes, and maintains steroid production with a high E2:P4 ratio in FWS cultures. PMID:23969977

The role of proton shuttling mechanisms in solvent-free and catalyst-free acetalization reactions of imines.

PubMed

Lillo, Victor J; Mansilla, Javier; Saá, José M

2018-06-06

Proton transfer is central to the understanding of chemical processes. More so in addition reactions of the type NuH + E → Nu-EH taking place under solvent-free and catalyst-free conditions. Herein we show that the addition of alcohols or amines (the NuH component) to imine derivatives (the E component), in 1 : 1 ratio, under solvent-free and catalyst-free conditions, are efficient methods to access N,O and N,N-acetal derivatives. In addition, computational studies reveal that they are catalyzed reactions involving two or even three NuH molecules operating in a cooperative manner as H-bonded NuH(NuH)nNuH associates (many body effects) in the transition state through a concerted proton shuttling mechanism (addition of alcohols) or stepwise proton shuttling mechanism (addition of amines), thereby facilitating the key proton transfer step.

Possibility of deuterium free water using as antitumoral means with reference to conditions of Martian expedition

NASA Astrophysics Data System (ADS)

Sinyak, Y.; Turusov, V.; Grigroriev, A.; Yaridze, D.; Gaidadimov, V.; Antoshina, E.; Gorkova, T.; Truhanova, L.

The interplanetary space flights, Martian program as an example, will take place under conditions of increasing radiation level on crew. The search of methods for a decrease of oncologic risk produced by irradiation of astronauts, is one of the major factors of a successful implementation of a flight program. One of such methods is a usage by crew of potable water with the reduced concentrations of a heavy stable isotope of hydrogen - deuterium, which can be obtained in the regenerative life support systems. The heavy water (D2O) has toxic properties, distorting biochemical reactions in the cell, inhibiting process of DNA replication. It can be presumed that the replacement of deuterium in the water for protium will result in normalization of cell metabolism, reparation will take place and this will lead to the inhibition of tumour development. In this study the water with a decreased by 65% of deuterium was used. Antitumour properties of D 2-free water were studied with transplantable Lewis lung carcinoma in BDF1 strain of mice. First results show that average time of appearance of the first nodules at the site of transplantation was 14 % longer in mice fed D 2-free water as compared to control. The tumour volume in the experimental group (decreased content of D2 ) was always lower than in the control. Statistically significant differences in the tumour volume were registered at the 13, 15, 23, 26 and 28 -th days after transplantation. Inhibition of tumour growth was equal to 100% and 51% at the 5 th and 15-th days after- transplantation respectively. Increase of life span in the experimental group was 10%. The results indicate that the use by astronauts of water with decreased content of deuterium may decrease the risk of oncological diseases under conditions of high radiation level in the flight to Mars.

Comparison of Different Physical Activity Measurement Methods in Adults Aged 45 to 64 Years Under Free-Living Conditions.

PubMed

Lipert, Anna; Jegier, Anna

2017-07-01

To compare physical activity (PA) measured by 4 methods in adults under free-living conditions in relation to selected demographic and anthropometric variables. Cohort study. Department of Sports Medicine. Clinically healthy men (81) and women (69) aged 45 to 64 years. Physical activity monitoring for 7 consecutive days under free-living conditions by pedometer (P) and accelerometer (A) simultaneously and PA questionnaires: International Physical Activity Questionnaire (IPAQ) and Seven-Day Physical Activity Questionnaire Recall (SDPAR) completed after the 7-day PA. Comparison of PA measured by pedometer, IPAQ, and SDPAR with accelerometer with regard to age, body mass, gender, and obesity type. Total energy expenditure (EE) by IPAQ was higher than A (P < 0.001) in both groups regardless of age, body mass, or obesity type. Mean EE value by P was greater than A (P < 0.001) in central-obesity males and lower than A (P < 0.001) in central-obesity females. There were differences in step counts in women, unnoticed in men. SDPAR overestimated total EE in gynoid-obesity males and in central-obesity females compared with A. Ninety-five percent CI was the largest around IPAQ compared with P and SDPAR, with SDPAR showing the best agreement with A. Body mass and obesity type influenced PA measurements. To monitor PA, it is recommended to use pedometer in normal bodyweight and overweight groups while accelerometer is advisable in obese subjects. A combined approach of objective and subjective PA monitoring tools is preferable.

Balancing consumer protection and scientific integrity in the face of uncertainty: the example of gluten-free foods.

PubMed

McCabe, Margaret Sova

2010-01-01

In 2009, gluten-free foods were not only "hot" in the marketplace, several countries, including the United States, continued efforts to define gluten-free and appropriate labeling parameters. The regulatory process illuminates how difficult regulations based on safe scientific thresholds can be for regulators, manufacturers and consumers. This article analyzes the gluten-free regulatory landscape, challenges to defining a safe gluten threshold, and how consumers might need more label information beyond the term "gluten-free." The article includes an overview of international gluten-free regulations, the Food and Drug Administration (FDA) rulemaking process, and issues for consumers.

Equilibrium Droplets on Deformable Substrates: Equilibrium Conditions.

PubMed

Koursari, Nektaria; Ahmed, Gulraiz; Starov, Victor M

2018-05-15

Equilibrium conditions of droplets on deformable substrates are investigated, and it is proven using Jacobi's sufficient condition that the obtained solutions really provide equilibrium profiles of both the droplet and the deformed support. At the equilibrium, the excess free energy of the system should have a minimum value, which means that both necessary and sufficient conditions of the minimum should be fulfilled. Only in this case, the obtained profiles provide the minimum of the excess free energy. The necessary condition of the equilibrium means that the first variation of the excess free energy should vanish, and the second variation should be positive. Unfortunately, the mentioned two conditions are not the proof that the obtained profiles correspond to the minimum of the excess free energy and they could not be. It is necessary to check whether the sufficient condition of the equilibrium (Jacobi's condition) is satisfied. To the best of our knowledge Jacobi's condition has never been verified for any already published equilibrium profiles of both the droplet and the deformable substrate. A simple model of the equilibrium droplet on the deformable substrate is considered, and it is shown that the deduced profiles of the equilibrium droplet and deformable substrate satisfy the Jacobi's condition, that is, really provide the minimum to the excess free energy of the system. To simplify calculations, a simplified linear disjoining/conjoining pressure isotherm is adopted for the calculations. It is shown that both necessary and sufficient conditions for equilibrium are satisfied. For the first time, validity of the Jacobi's condition is verified. The latter proves that the developed model really provides (i) the minimum of the excess free energy of the system droplet/deformable substrate and (ii) equilibrium profiles of both the droplet and the deformable substrate.

Assessment of season-dependent body condition scores in relation to faecal glucocorticoid metabolites in free-ranging Asian elephants

PubMed Central

Pokharel, Sanjeeta Sharma; Seshagiri, Polani B.

2017-01-01

Abstract We studied seasonal and annual changes in visual body condition scores (BCSs), and assessed how these scores were related to levels of faecal glucocorticoid metabolites (fGCMs) in free-ranging Asian elephants (Elephas maximus) in the seasonally dry tropical forests of the Mysore and Nilgiri Elephant Reserves in southern India. We assessed the animals’ BCS visually on a scale of 1 to 5; where 1 represents a very thin and 5 represents a very fat elephant. To understand the influence of seasonality on BCS, we sampled the population during dry (n = 398) and wet seasons (n = 255) of 2013 and 2015 while, for annual changes in BCS, we sampled nine free-ranging adult females from different family groups that had been repeatedly sighted over seven years. To evaluate the influence of body condition on fGCM, 307 faecal samples were collected from 261 different elephants and were analysed. As a parameter of adrenocortical activity, and thus stress, fGCM was measured (μg/g) in the ethanol-extracted samples using a group-specific 11-oxoaetiocholanolone EIA (antibody raised against 11-oxoaetiocholanolone-17-CMO:BSA and biotinylated-11-oxoaetiocholanolone as a label). Effect of age and season on BCS in relation to fGCM was also studied. A seasonal shift in BCS was observed as expected, i.e. individuals with low BCS were more frequent during the dry season when compared with the wet season. Concentrations of fGCM were highest in individuals with lowest BCS (BCS 1) and then significantly declined till BCS 3. fGCM levels were almost comparable for BCS 3, 4 and 5. This pattern was more conspicuous in female than in male elephants. Season-dependent BCS, hence, reflect the stress status as measured by fGCM, especially in female Asian elephants. This could be used as an important non-invasive approach to monitor the physiological health of free-ranging elephant populations. PMID:28721215

Two-way shape memory behavior of semi-crystalline elastomer under stress-free condition

NASA Astrophysics Data System (ADS)

Qian, Chen; Dong, Yubing; Zhu, Yaofeng; Fu, Yaqin

2016-08-01

Semi-crystalline shape memory polymers exhibit two-way shape memory effect (2W-SME) under constant stresses through crystallization-induced elongation upon cooling and melting-induced constriction upon heating. The applied constant stress influenced the prediction and usability of 2W-SME in practical applications without any external force. Here the reversible shape transition in EVA-shaped memory polymer was quantitative analyzed under a suitable temperature range and external stress-free condition. The fraction of reversible strain increased with increasing upper temperature (T high) within the temperature range and reached the maximum value of 13.62% at 70 °C. However, reversible strain transition was almost lost when T high exceeded 80 °C because of complete melting of crystalline scaffold, known as the latent recrystallization template. The non-isothermal annealing of EVA 2W-SMP under changing circulating temperatures was confirmed. Moreover, the orientation of crystallization was retained at high temperatures. These findings may contribute to design an appropriate shape memory protocol based on application-specific requirements.

NNP-Type Pincer Imidazolylphosphine Ruthenium Complexes: Efficient Base-Free Hydrogenation of Aromatic and Aliphatic Nitriles under Mild Conditions.

PubMed

Adam, Rosa; Alberico, Elisabetta; Baumann, Wolfgang; Drexler, Hans-Joachim; Jackstell, Ralf; Junge, Henrik; Beller, Matthias

2016-03-24

A series of seven novel N(Im)N(H)P-type pincer imidazolylphosphine ruthenium complexes has been synthesized and fully characterized. The use of hydrogenation of benzonitrile as a benchmark test identified [RuHCl(CO)(N(Im)N(H) P(tBu))] as the most active catalyst. With its stable Ru-BH4 analogue, in which chloride is replaced by BH4, a broad range of (hetero)aromatic and aliphatic nitriles, including industrially interesting adiponitrile, has been hydrogenated under mild and base-free conditions. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Open Loop Heat Pipe Radiator Having a Free-Piston for Wiping Condensed Working Fluid

NASA Technical Reports Server (NTRS)

Weinstein, Leonard M. (Inventor)

2015-01-01

An open loop heat pipe radiator comprises a radiator tube and a free-piston. The radiator tube has a first end, a second end, and a tube wall, and the tube wall has an inner surface and an outer surface. The free-piston is enclosed within the radiator tube and is capable of movement within the radiator tube between the first and second ends. The free-piston defines a first space between the free-piston, the first end, and the tube wall, and further defines a second space between the free-piston, the second end, and the tube wall. A gaseous-state working fluid, which was evaporated to remove waste heat, alternately enters the first and second spaces, and the free-piston wipes condensed working fluid from the inner surface of the tube wall as the free-piston alternately moves between the first and second ends. The condensed working fluid is then pumped back to the heat source.

The Effect of Belief in Free Will on Prejudice

PubMed Central

Zhao, Xian; Liu, Li; Zhang, Xiao-xiao; Shi, Jia-xin; Huang, Zhen-wei

2014-01-01

The current research examined the role of the belief in free will on prejudice across Han Chinese and white samples. Belief in free will refers to the extent to which people believe human beings truly have free will. In Study 1, the beliefs of Han Chinese people in free will were measured, and their social distances from the Tibetan Chinese were used as an index of ethnic prejudice. The results showed that the more that Han Chinese endorsed the belief in free will, the less that they showed prejudice against the Tibetan Chinese. In Study 2, the belief of the Han Chinese in free will was manipulated, and their explicit feelings towards the Uyghur Chinese were used as an indicator of ethnic prejudice. The results showed that the participants in the condition of belief in free will reported less prejudice towards Uyghur Chinese compared to their counterparts in the condition of disbelief in free will. In Study 3, white peoples’ belief in free will was manipulated, and their pro-black attitudes were measured as an indirect indicator of racial prejudice. The results showed that, compared to the condition of disbelief in free will, the participants who were primed by a belief in free will reported stronger pro-black attitudes. These three studies suggest that endorsement of the belief in free will can lead to decreased ethnic/racial prejudice compared to denial of the belief in free will. The theoretical and practical implications are discussed. PMID:24622280

The effect of belief in free will on prejudice.

PubMed

Zhao, Xian; Liu, Li; Zhang, Xiao-xiao; Shi, Jia-xin; Huang, Zhen-wei

2014-01-01

The current research examined the role of the belief in free will on prejudice across Han Chinese and white samples. Belief in free will refers to the extent to which people believe human beings truly have free will. In Study 1, the beliefs of Han Chinese people in free will were measured, and their social distances from the Tibetan Chinese were used as an index of ethnic prejudice. The results showed that the more that Han Chinese endorsed the belief in free will, the less that they showed prejudice against the Tibetan Chinese. In Study 2, the belief of the Han Chinese in free will was manipulated, and their explicit feelings towards the Uyghur Chinese were used as an indicator of ethnic prejudice. The results showed that the participants in the condition of belief in free will reported less prejudice towards Uyghur Chinese compared to their counterparts in the condition of disbelief in free will. In Study 3, white peoples' belief in free will was manipulated, and their pro-black attitudes were measured as an indirect indicator of racial prejudice. The results showed that, compared to the condition of disbelief in free will, the participants who were primed by a belief in free will reported stronger pro-black attitudes. These three studies suggest that endorsement of the belief in free will can lead to decreased ethnic/racial prejudice compared to denial of the belief in free will. The theoretical and practical implications are discussed.

The inhibitory effects of free ammonia on ammonia oxidizing bacteria and nitrite oxidizing bacteria under anaerobic condition.

PubMed

Qian, Wenting; Peng, Yongzhen; Li, Xiyao; Zhang, Qiong; Ma, Bin

2017-11-01

The free ammonia (FA) inhibition on ammonia oxidizing bacteria (AOB) and nitrite oxidizing bacteria (NOB) under anaerobic condition was investigated in this study. The results indicated that NOB was more sensitive to the FA anaerobic treatment than AOB. The FA anaerobic inhibition on nitrifier gradually heightened with the increase of FA concentration. Accompanied with FA concentration increase from 0 to 16.82mgNH 3 -N·L -1 (the highest concentration adopted in this study), the activity of AOB reduced by 15.9%, while NOB decreased by 29.2%. After FA anaerobic treatment, nitrite was accumulated during nitrification. However, the nitrite accumulation disappeared on the sixth cycle of activity recovery tests with excessive aeration. Based on this result, a novel strategy for achieving nitritation is proposed, which involves recirculating a portion of the activated sludge through a side-line sludge treatment unit, where the sludge is subjected to treatment with FA under anaerobic condition. Copyright © 2017 Elsevier Ltd. All rights reserved.

Global stability and tumor clearance conditions for a cancer chemotherapy system

NASA Astrophysics Data System (ADS)

Valle, Paul A.; Starkov, Konstantin E.; Coria, Luis N.

2016-11-01

In this paper we study the global dynamics of a cancer chemotherapy system presented by de Pillis et al. (2007). This mathematical model describes the interaction between tumor cells, effector-immune cells, circulating lymphocytes and chemotherapy treatment. By applying the localization method of compact invariant sets, we find lower and upper bounds for these three cells populations. Further, we define a bounded domain in R+,04 where all compact invariant sets of the system are located and provide conditions under which this domain is positively invariant. We apply LaSalle's invariance principle and one result concerning two-dimensional competitive systems in order to derive sufficient conditions for tumor clearance and global asymptotic stability of the tumor-free equilibrium point. These conditions are computed by using bounds of the localization domain and they are given in terms of the chemotherapy treatment. Finally, we perform numerical simulations in order to illustrate our results.

Effect of ascorbic acid (vitamin C) on the ESR spectra of the red and black hair: pheomelanin free radicals are not always present in red hair.

PubMed

Chikvaidze, Eduard; Topeshashvili, Maia

2015-12-01

Increased incidence of melanoma in the population with red hair is conditioned by synthesis of pheomelanin pigments in the skin and their phototoxic properties. The recent research has shown that free radicals of pheomelanin are produced not only by the influence of UV irradiation, but also in UV-independent pathways of oxidative stress. It has been ascertained, that the color of the hair is not always determinant of the amount of pheolemanin radicals in red hair. Therefore, in order to evaluate the risk of melanoma in different individuals, it is necessary to define the amount of free radicals of pheomelanin in red hair using ESR spectroscopy method. Besides, it is very important to find effective antioxidant, capable of neutralizing free radicals of pheomelanin. It was proved that ascorbic acid neutralizes free radicals of pheomelanin very effectively. The main goal of our research was to define the presumably optimal concentration of ascorbic acid as an antioxidant and study the kinetics of the influence of this concentration on red and black hair. It has been found out, that ascorbic acid influences the free radicals of red and black hair, and its appropriate optimal concentration is 10 mM. The obtained results can be considered in dermatology and cosmetology. Copyright © 2015 John Wiley & Sons, Ltd.

Linear models of coregionalization for multivariate lattice data: Order-dependent and order-free cMCARs.

PubMed

MacNab, Ying C

2016-08-01

This paper concerns with multivariate conditional autoregressive models defined by linear combination of independent or correlated underlying spatial processes. Known as linear models of coregionalization, the method offers a systematic and unified approach for formulating multivariate extensions to a broad range of univariate conditional autoregressive models. The resulting multivariate spatial models represent classes of coregionalized multivariate conditional autoregressive models that enable flexible modelling of multivariate spatial interactions, yielding coregionalization models with symmetric or asymmetric cross-covariances of different spatial variation and smoothness. In the context of multivariate disease mapping, for example, they facilitate borrowing strength both over space and cross variables, allowing for more flexible multivariate spatial smoothing. Specifically, we present a broadened coregionalization framework to include order-dependent, order-free, and order-robust multivariate models; a new class of order-free coregionalized multivariate conditional autoregressives is introduced. We tackle computational challenges and present solutions that are integral for Bayesian analysis of these models. We also discuss two ways of computing deviance information criterion for comparison among competing hierarchical models with or without unidentifiable prior parameters. The models and related methodology are developed in the broad context of modelling multivariate data on spatial lattice and illustrated in the context of multivariate disease mapping. The coregionalization framework and related methods also present a general approach for building spatially structured cross-covariance functions for multivariate geostatistics. © The Author(s) 2016.

12 CFR 914.1 - Regulatory Report defined.

Code of Federal Regulations, 2010 CFR

2010-01-01

... Banks and Banking FEDERAL HOUSING FINANCE BOARD GOVERNANCE AND MANAGEMENT OF THE FEDERAL HOME LOAN BANKS DATA AVAILABILITY AND REPORTING § 914.1 Regulatory Report defined. (a) Definition. Regulatory Report means any report of raw or summary data needed to evaluate the safe and sound condition and operations...

Accumulation of the Cyclobutane Thymine Dimer in Defined Sequences of Free and Nucleosomal DNA

DTIC Science & Technology

2013-08-01

cyclobutane dimer in a single-stranded vector , Proc. Natl. Acad. Sci. U. S. A., 1988, 85, 8141–8145. 11 C. A. Smith, M. Wang, N. Jiang, L. Che, X. Zhao and...J.-S. Taylor, Mutation spectra of M13 vectors containing site-specific cis–syn, trans–syn-I, (6-4), and Dewar pyrimi- done photoproducts of thymidylyl...Bypass of a site-specific cis–syn thymine dimer in a SV40 vector during in vitro replication by HeLa and XPV cell-free extracts, Biochemistry, 1998

Electron-beam conditioning by thomson scattering.

PubMed

Schroeder, C B; Esarey, E; Leemans, W P

2004-11-05

A method is proposed for conditioning electron beams via Thomson scattering. The conditioning provides a quadratic correlation between the electron energy deviation and the betatron amplitude of the electrons, which results in enhanced gain in free-electron lasers. Quantum effects imply conditioning must occur at high laser fluence and moderate electron energy. Conditioning of x-ray free-electron lasers should be achievable with present laser technology, leading to significant size and cost reductions of these large-scale facilities.

Covalent protein-oligonucleotide conjugates by copper-free click reaction

PubMed Central

Khatwani, Santoshkumar L.; Mullen, Daniel G.; Hast, Michael A.; Beese, Lorena S.; Distefano, Mark D.; Taton, T. Andrew

2013-01-01

Covalent protein-oligodeoxynucleotide (protein-ODN) conjugates are useful in a number of biological applications, but synthesizing discrete conjugates—where the connection between the two components is at a defined location in both the protein and the ODN—under mild conditions with significant yield can be a challenge. In this article, we demonstrate a strategy for synthesizing discrete protein-ODN conjugates using strain-promoted azide-alkyne [3+2] cycloaddition (SPAAC, a copper-free “click” reaction). Azide-functionalized proteins, prepared by enzymatic prenylation of C-terminal CVIA tags with synthetic azidoprenyl diphosphates, were “clicked” to ODNs that had been modified with a strained dibenzocyclooctyne (DIBO-ODN). The resulting protein-ODN conjugates were purified and characterized by size-exclusion chromatography and gel electrophoresis. We find that the yields and reaction times of the SPAAC bioconjugation reactions are comparable to those previously reported for copper-catalyzed azide-alkyne [3+2] cycloaddition (CuAAC) bioconjugation, but require no catalyst. The same SPAAC chemistry was used to immobilize azide-modified proteins onto surfaces, using surface-bound DIBO-ODN as a heterobifunctional linker. Cu-free click bioconjugation of proteins to ODNs is a simple and versatile alternative to Cu-catalyzed click methods. PMID:22682299

A Model-Free Diagnostic for Single-Peakedness of Item Responses Using Ordered Conditional Means.

PubMed

Polak, Marike; de Rooij, Mark; Heiser, Willem J

2012-09-01

In this article we propose a model-free diagnostic for single-peakedness (unimodality) of item responses. Presuming a unidimensional unfolding scale and a given item ordering, we approximate item response functions of all items based on ordered conditional means (OCM). The proposed OCM methodology is based on Thurstone & Chave's (1929) criterion of irrelevance, which is a graphical, exploratory method for evaluating the "relevance" of dichotomous attitude items. We generalized this criterion to graded response items and quantified the relevance by fitting a unimodal smoother. The resulting goodness-of-fit was used to determine item fit and aggregated scale fit. Based on a simulation procedure, cutoff values were proposed for the measures of item fit. These cutoff values showed high power rates and acceptable Type I error rates. We present 2 applications of the OCM method. First, we apply the OCM method to personality data from the Developmental Profile; second, we analyze attitude data collected by Roberts and Laughlin (1996) concerning opinions of capital punishment.

Amputee skin condition: occlusion, stratum corneum hydration and free amino acid levels.

PubMed

Visscher, Marty O; Robinson, Marisa; Fugit, Benetta; Rosenberg, Richard J; Hoath, Steven B; Randall Wickett, R

2011-03-01

Patients with a prosthetic limb report negative skin effects, including irritation, rash and chafing, which can lead to infection, discomfort and reduced wear time to significantly impact normal activities. The aims were to examine the epidermal integrity (transepidermal water loss, TEWL), stratum corneum (SC) hydration [moisture accumulation rate (MAT)], friction and biomechanical properties in active below the knee amputees and to determine the effects of an inert sock liner on skin condition. The liner reduced hydration, TEWL and friction and increased elasticity versus the amputee's conventional skin care methods. Residual limb TEWL was increased and MAT was reduced versus the contralateral normal skin. In a second study, we hypothesized that complete occlusion would decrease free amino acids (FAA) and quantified them by high performance liquid chromatography in an adult volar forearm model. Occlusion with a water vapor impermeable wet dressing led to increased TEWL, erythema and dryness and reduced MAT versus normal skin, comparable to the results in the amputees. The FAA levels were significantly reduced for the occluded sites. The results suggest that residual limb occlusion in amputees may block the formation of FAA in the upper SC. Therapies based on replacement of water binding FAAs, may alleviate the consequences of long-term occlusion.

Circular free-electron laser

DOEpatents

Brau, Charles A.; Kurnit, Norman A.; Cooper, Richard K.

1984-01-01

A high efficiency, free electron laser utilizing a circular relativistic electron beam accelerator and a circular whispering mode optical waveguide for guiding optical energy in a circular path in the circular relativistic electron beam accelerator such that the circular relativistic electron beam and the optical energy are spatially contiguous in a resonant condition for free electron laser operation. Both a betatron and synchrotron are disclosed for use in the present invention. A free electron laser wiggler is disposed around the circular relativistic electron beam accelerator for generating a periodic magnetic field to transform energy from the circular relativistic electron beam to optical energy.

Enamel and dentin bond strength following gaseous ozone application.

PubMed

Cadenaro, Milena; Delise, Chiara; Antoniollo, Francesca; Navarra, Ottavia Chiara; Di Lenarda, Roberto; Breschi, Lorenzo

2009-08-01

To evaluate the effects of gaseous ozone application on enamel and dentin bond strength produced by two self-etching adhesive systems. The shear bond strength test was conducted to assess adhesion on enamel (protocol 1), while the microtensile bond strength test was performed on dentin (protocol 2). Protocol 1: 96 bovine incisors were randomly divided into 4 groups, and enamel surfaces were bonded in accordance with the following treatments: (1E) ozone + Clearfil Protect Bond; (2E) Clearfil Protect Bond (control); (3E) ozone + Xeno III; (4E) Xeno III (control). Ozone gas was applied for 80 s. Shear bond strength was measured with a universal testing machine. Protocol 2: 40 noncarious human molars were selected. Middle/deep dentin was exposed and bonded in accordance with the following treatments: (1D) ozone+Clearfil Protect Bond; (2D) Clearfil Protect Bond (control); (3D) ozone+Xeno III (4D) Xeno III (control). Four-mm-thick buildups were built on the adhesives, then specimens were sectioned in accordance with the nontrimming technique. Specimens were stressed until failure occurred, and failure modes were analyzed. Shear bond and microtensile bond strength data were analyzed using two-way ANOVA and Tukey's post-hoc test. No statistical differences were found between ozone treated specimens and controls, neither on enamel nor on dentin irrespective of the tested adhesive. Clearfil Protect Bond showed higher bond strength to enamel than Xeno III, irrespective of the ozone treatment (p < 0.05). The use of ozone gas to disinfect the cavity before placing a restoration had no influence on immediate enamel and dentin bond strength.

Reusable ionic liquid-catalyzed oxidative coupling of azoles and benzylic compounds via sp(3) C-N bond formation under metal-free conditions.

PubMed

Liu, Wenbo; Liu, Chenjiang; Zhang, Yonghong; Sun, Yadong; Abdukadera, Ablimit; Wang, Bin; Li, He; Ma, Xuecheng; Zhang, Zengpeng

2015-07-14

The heterocyclic ionic liquid-catalyzed direct oxidative amination of benzylic sp(3) C-H bonds via intermolecular sp(3) C-N bond formation for the synthesis of N-alkylated azoles under metal-free conditions is reported for the first time. The catalyst 1-butylpyridinium iodide can be recycled and reused with similar efficacies for at least eight cycles.

Does the use of a novel self-adhesive flowable composite reduce nanoleakage?

PubMed

Naga, Abeer Abo El; Yousef, Mohammed; Ramadan, Rasha; Fayez Bahgat, Sherif; Alshawwa, Lana

2015-01-01

The aim of the study reported here was to evaluate the performance of a self-adhesive flowable composite and two self-etching adhesive systems, when subjected to cyclic loading, in preventing the nanoleakage of Class V restorations. Wedge-shape Class V cavities were prepared (4×2×2 mm [length × width × depth]) on the buccal surfaces of 90 sound human premolars. Cavities were divided randomly into three groups (n=30) according to the used adhesive (Xeno(®) V [self-etching adhesive system]) and BOND-1(®) SF (solvent-free self-etching adhesive system) in conjunction with Artiste(®) Nano Composite resin, and Fusio™ Liquid Dentin (self-adhesive flowable composite), consecutively. Each group was further divided into three subgroups (n=10): (A) control, (B) subjected to occlusal cyclic loading (90N for 5,000 cycles), and (C) subjected to occlusal cyclic loading (90N for 10,000 cycles). Teeth then were coated with nail polish up to 1 mm from the interface, immersed in 50% silver nitrate solution for 24 hours and tested for nanoleakage using the environmental scanning electron microscopy and energy dispersive analysis X-ray analysis. Data were statistically analyzed using two-way analysis of variance and Tukey's post hoc tests (P≤0.05). The Fusio Liquid Dentin group showed statistically significant lower percentages of silver penetration (0.55 μ) compared with the BOND-1 SF (3.45 μ) and Xeno V (3.82 μ) groups, which were not statistically different from each other, as they both showed higher silver penetration. Under the test conditions, the self-adhesive flowable composite provided better sealing ability. Aging of the two tested adhesive systems, as a function of cyclic loading, increased nanoleakage.

Does the use of a novel self-adhesive flowable composite reduce nanoleakage?

PubMed Central

Naga, Abeer Abo El; Yousef, Mohammed; Ramadan, Rasha; Fayez Bahgat, Sherif; Alshawwa, Lana

2015-01-01

Objective The aim of the study reported here was to evaluate the performance of a self-adhesive flowable composite and two self-etching adhesive systems, when subjected to cyclic loading, in preventing the nanoleakage of Class V restorations. Methods Wedge-shape Class V cavities were prepared (4×2×2 mm [length × width × depth]) on the buccal surfaces of 90 sound human premolars. Cavities were divided randomly into three groups (n=30) according to the used adhesive (Xeno® V [self-etching adhesive system]) and BOND-1® SF (solvent-free self-etching adhesive system) in conjunction with Artiste® Nano Composite resin, and Fusio™ Liquid Dentin (self-adhesive flowable composite), consecutively. Each group was further divided into three subgroups (n=10): (A) control, (B) subjected to occlusal cyclic loading (90N for 5,000 cycles), and (C) subjected to occlusal cyclic loading (90N for 10,000 cycles). Teeth then were coated with nail polish up to 1 mm from the interface, immersed in 50% silver nitrate solution for 24 hours and tested for nanoleakage using the environmental scanning electron microscopy and energy dispersive analysis X-ray analysis. Data were statistically analyzed using two-way analysis of variance and Tukey’s post hoc tests (P≤0.05). Results The Fusio Liquid Dentin group showed statistically significant lower percentages of silver penetration (0.55 μ) compared with the BOND-1 SF (3.45 μ) and Xeno V (3.82 μ) groups, which were not statistically different from each other, as they both showed higher silver penetration. Conclusion Under the test conditions, the self-adhesive flowable composite provided better sealing ability. Aging of the two tested adhesive systems, as a function of cyclic loading, increased nanoleakage. PMID:25848318

Feasibility of pig and human-derived aortic valve interstitial cells seeding on fixative-free decellularized animal pericardium.

PubMed

Santoro, Rosaria; Consolo, Filippo; Spiccia, Marco; Piola, Marco; Kassem, Samer; Prandi, Francesca; Vinci, Maria Cristina; Forti, Elisa; Polvani, Gianluca; Fiore, Gianfranco Beniamino; Soncini, Monica; Pesce, Maurizio

2016-02-01

Glutaraldehyde-fixed pericardium of animal origin is the elective material for the fabrication of bio-prosthetic valves for surgical replacement of insufficient/stenotic cardiac valves. However, the pericardial tissue employed to this aim undergoes severe calcification due to chronic inflammation resulting from a non-complete immunological compatibility of the animal-derived pericardial tissue resulting from failure to remove animal-derived xeno-antigens. In the mid/long-term, this leads to structural deterioration, mechanical failure, and prosthesis leaflets rupture, with consequent need for re-intervention. In the search for novel procedures to maximize biological compatibility of the pericardial tissue into immunocompetent background, we have recently devised a procedure to decellularize the human pericardium as an alternative to fixation with aldehydes. In the present contribution, we used this procedure to derive sheets of decellularized pig pericardium. The decellularized tissue was first tested for the presence of 1,3 α-galactose (αGal), one of the main xenoantigens involved in prosthetic valve rejection, as well as for mechanical tensile behavior and distensibility, and finally seeded with pig- and human-derived aortic valve interstitial cells. We demonstrate that the decellularization procedure removed the αGAL antigen, maintained the mechanical characteristics of the native pig pericardium, and ensured an efficient surface colonization of the tissue by animal- and human-derived aortic valve interstitial cells. This establishes, for the first time, the feasibility of fixative-free pericardial tissue seeding with valve competent cells for derivation of tissue engineered heart valve leaflets. © 2015 Wiley Periodicals, Inc.

Free love? On the relation between belief in free will, determinism, and passionate love.

PubMed

Boudesseul, Jordane; Lantian, Anthony; Cova, Florian; Bègue, Laurent

2016-11-01

Is love possible if we are not free? Some philosophers consider that true love is necessarily free, while others think that the nature of love makes it incompatible with a certain type of freedom. Here, we explored the relationship between feelings of passionate love, belief in free will and determinism across three online studies. In Study 1 (N=257), participants who believed strongly in free will (or determinism) expressed stronger passionate love. In Study 2 (N=305), we again found a positive association between belief in free will (or determinism) and passionate love, although the passionate love-determinism relationship seems more conditional. Finally, Study 3 (N=309) confirmed the relationship between belief in free will and passionate love but not between belief in determinism and passionate love. These findings, along with a meta-analysis, suggest that both beliefs in free will and determinism are compatible with passionate love. Copyright © 2016 Elsevier Inc. All rights reserved.

Modelling of 10 Gbps Free Space Optics Communication Link Using Array of Receivers in Moderate and Harsh Weather Conditions

NASA Astrophysics Data System (ADS)

Gupta, Amit; Shaina, Nagpal

2017-08-01

Intersymbol interference and attenuation of signal are two major parameters affecting the quality of transmission in Free Space Optical (FSO) Communication link. In this paper, the impact of these parameters on FSO communication link is analysed for delivering high-quality data transmission. The performance of the link is investigated under the influence of amplifier in the link. The performance parameters of the link like minimum bit error rate, received signal power and Quality factor are examined by employing erbium-doped fibre amplifier in the link. The effects of amplifier are visualized with the amount of received power. Further, the link is simulated for moderate weather conditions at various attenuation levels on transmitted signal. Finally, the designed link is analysed in adverse weather conditions by using high-power laser source for optimum performance.

Donor free radical explosive composition

DOEpatents

Walker, Franklin E. [15 Way Points Rd., Danville, CA 94526; Wasley, Richard J. [4290 Colgate Way, Livermore, CA 94550

1980-04-01

An improved explosive composition is disclosed and comprises a major portion of an explosive having a detonation velocity between about 1500 and 10,000 meters per second and a minor amount of a donor additive comprising an organic compound or mixture of organic compounds capable of releasing low molecular weight free radicals or ions under mechanical or electrical shock conditions and which is not an explosive, or an inorganic compound or mixture of inorganic compounds capable of releasing low molecular weight free radicals or ions under mechanical or electrical shock conditions and selected from ammonium or alkali metal persulfates.

High-performance MgB2 superconducting wires for use under liquid-helium-free conditions fabricated using an internal Mg diffusion process

NASA Astrophysics Data System (ADS)

Ye, ShuJun; Song, Minghui; Matsumoto, Akiyoshi; Togano, Kazumasa; Takeguchi, Masaki; Ohmura, Takahito; Kumakura, Hiroaki

2013-12-01

MgB2 has a superconducting transition temperature (Tc) of 39 K, which is much higher than that for practical metallic superconductors. Thus, it is hoped that MgB2 can not only replace metallic superconductors, but can be used under liquid-helium-free conditions, for example, at temperatures of 10-20 K that can easily be achieved using cryocooling systems. However, to date, the reported critical current density (Jc) for MgB2 wires is not high enough for large-scale applications in liquid-helium-free conditions. In the present study, successful fabrication of high-performance MgB2 superconducting wires was carried out using an internal Mg diffusion (IMD) process, involving a p-dimethylbenzene (C8H10) pre-treatment of carbon-coated B powder with nanometer-sized particles. The resulting wires exhibited the highest ever Jc of 1.2 × 105 A cm-2 at 4.2 K and 10 T, and an engineering critical current density (Je) of about 1 × 104 A cm-2. Not only in 4.2 K, but also in 10 K, the Jc values for the wires fabricated in the present study are in fact higher than that for Nb-Ti wires at 4.2 K for the magnetic fields at which the measurements were carried out. At 20 K and 5 T, the Jc and Je were about 7.6 × 105 A cm-2 and 5.3 × 103 A cm-2, respectively, which are the highest values reported for MgB2 wires to date. The results of a detailed microstructural analysis suggested that the main reason for the superior electrical performance was the high density of the MgB2 layer rather than just the small grain size, and that the critical current could be further increased by suitable control of the microstructure. These high-performance IMD-processed MgB2 wires are thus promising superconductors for applications such as magnetic resonance imaging and maglev trains that can operate under liquid-helium-free conditions.

Refined boundary conditions on the free surface of an elastic half-space taking into account non-local effects.

PubMed

Chebakov, R; Kaplunov, J; Rogerson, G A

2016-02-01

The dynamic response of a homogeneous half-space, with a traction-free surface, is considered within the framework of non-local elasticity. The focus is on the dominant effect of the boundary layer on overall behaviour. A typical wavelength is assumed to considerably exceed the associated internal lengthscale. The leading-order long-wave approximation is shown to coincide formally with the 'local' problem for a half-space with a vertical inhomogeneity localized near the surface. Subsequent asymptotic analysis of the inhomogeneity results in an explicit correction to the classical boundary conditions on the surface. The order of the correction is greater than the order of the better-known correction to the governing differential equations. The refined boundary conditions enable us to evaluate the interior solution outside a narrow boundary layer localized near the surface. As an illustration, the effect of non-local elastic phenomena on the Rayleigh wave speed is investigated.

The experimental electron mean-free-path in Si under typical (S)TEM conditions.

PubMed

Potapov, P L

2014-12-01

The electron mean-free-path in Si was measured by EELS using the test structure with the certified dimensions as a calibration standard. In a good agreement with the previous CBED measurements, the mean-free-path is 150nm for 200keV and 179nm for 300keV energy of primary electrons at large collection angles. These values are accurately predicted by the model of Iakoubovskii et al. while the model of Malis et al. incorporated in common microscopy software underestimates the mean-free-path by 15% at least. Correspondingly, the thickness of TEM samples reported in many studies of the Si-based materials last decades might be noticeably underestimated. Copyright © 2014 Elsevier B.V. All rights reserved.

Metabolism of vertebrate amino sugars with N-glycolyl groups: mechanisms underlying gastrointestinal incorporation of the non-human sialic acid xeno-autoantigen N-glycolylneuraminic acid.

PubMed

Banda, Kalyan; Gregg, Christopher J; Chow, Renee; Varki, Nissi M; Varki, Ajit

2012-08-17

Although N-acetyl groups are common in nature, N-glycolyl groups are rare. Mammals express two major sialic acids, N-acetylneuraminic acid and N-glycolylneuraminic acid (Neu5Gc). Although humans cannot produce Neu5Gc, it is detected in the epithelial lining of hollow organs, endothelial lining of the vasculature, fetal tissues, and carcinomas. This unexpected expression is hypothesized to result via metabolic incorporation of Neu5Gc from mammalian foods. This accumulation has relevance for diseases associated with such nutrients, via interaction with Neu5Gc-specific antibodies. Little is known about how ingested sialic acids in general and Neu5Gc in particular are metabolized in the gastrointestinal tract. We studied the gastrointestinal and systemic fate of Neu5Gc-containing glycoproteins (Neu5Gc-glycoproteins) or free Neu5Gc in the Neu5Gc-free Cmah(-/-) mouse model. Ingested free Neu5Gc showed rapid absorption into the circulation and urinary excretion. In contrast, ingestion of Neu5Gc-glycoproteins led to Neu5Gc incorporation into the small intestinal wall, appearance in circulation at a steady-state level for several hours, and metabolic incorporation into multiple peripheral tissue glycoproteins and glycolipids, thus conclusively proving that Neu5Gc can be metabolically incorporated from food. Feeding Neu5Gc-glycoproteins but not free Neu5Gc mimics the human condition, causing tissue incorporation into human-like sites in Cmah(-/-) fetal and adult tissues, as well as developing tumors. Thus, glycoproteins containing glycosidically linked Neu5Gc are the likely dietary source for human tissue accumulation, and not the free monosaccharide. This human-like model can be used to elucidate specific mechanisms of Neu5Gc delivery from the gut to tissues, as well as general mechanisms of metabolism of ingested sialic acids.

Free-space optical channel simulator for weak-turbulence conditions.

PubMed

Bykhovsky, Dima

2015-11-01

Free-space optical (FSO) communication may be severely influenced by the inevitable turbulence effect that results in channel gain fluctuations and fading. The objective of this paper is to provide a simple and effective simulator of the weak-turbulence FSO channel that emulates the influence of the temporal covariance effect. Specifically, the proposed model is based on lognormal distributed samples with a corresponding correlation time. The simulator is based on the solution of the first-order stochastic differential equation (SDE). The results of the provided SDE analysis reveal its efficacy for turbulent channel modeling.

An efficient synthesis of 3,4-Dihydropyrimidin-2(1H)-ones and thiones catalyzed by a novel Brønsted acidic ionic liquid under solvent-free conditions.

PubMed

Zhang, Yonghong; Wang, Bin; Zhang, Xiaomei; Huang, Jianbin; Liu, Chenjiang

2015-02-26

We report here an efficient and green method for Biginelli condensation reaction of aldehydes, β-ketoesters and urea or thiourea catalyzed by Brønsted acidic ionic liquid [Btto][p-TSA] under solvent-free conditions. Compared to the classical Biginelli reaction conditions, the present method has the advantages of giving good yields, short reaction times, near room temperature conditions and the avoidance of the use of organic solvents and metal catalyst.

Instabilities of the force-free current configurations

NASA Astrophysics Data System (ADS)

Berseth, V.; Indenbom, M. V.; van der Beek, C. J.; Erb, A.; Walker, E.; Flükiger, R.; Benoit, W.

1996-03-01

Using the magneto-optic technique, it is shown that inductively induced force-free current configurations in high purity YBa2Cu3O7-δ single crystals become unstable above a certain well-defined amplitude and frequency of the variation of the applied perpendicular field.

A new way of setting the phases for cosmological multiscale Gaussian initial conditions

NASA Astrophysics Data System (ADS)

Jenkins, Adrian

2013-09-01

We describe how to define an extremely large discrete realization of a Gaussian white noise field that has a hierarchical structure and the property that the value of any part of the field can be computed quickly. Tiny subregions of such a field can be used to set the phase information for Gaussian initial conditions for individual cosmological simulations of structure formation. This approach has several attractive features: (i) the hierarchical structure based on an octree is particularly well suited for generating follow-up resimulation or zoom initial conditions; (ii) the phases are defined for all relevant physical scales in advance so that resimulation initial conditions are, by construction, consistent both with their parent simulation and with each other; (iii) the field can easily be made public by releasing a code to compute it - once public, phase information can be shared or published by specifying a spatial location within the realization. In this paper, we describe the principles behind creating such realizations. We define an example called Panphasia and in a companion paper by Jenkins and Booth (2013) make public a code to compute it. With 50 octree levels Panphasia spans a factor of more than 1015 in linear scale - a range that significantly exceeds the ratio of the current Hubble radius to the putative cold dark matter free-streaming scale. We show how to modify a code used for making cosmological and resimulation initial conditions so that it can take the phase information from Panphasia and, using this code, we demonstrate that it is possible to make good quality resimulation initial conditions. We define a convention for publishing phase information from Panphasia and publish the initial phases for several of the Virgo Consortium's most recent cosmological simulations including the 303 billion particle MXXL simulation. Finally, for reference, we give the locations and properties of several dark matter haloes that can be resimulated within these

A Measurement-Free Approach to Conditioning.

DTIC Science & Technology

1987-11-01

y) Iv TO) * (.S Is2DUEO LUNG CONGESTION Z2b 2Ez2] EQI(bZ)-fl.3...U where n0 is given in (2.7) and - :1:,0 vb12 ~ NOLGLIOF ISESESTAE 1 0, =%, 5M( i...uniquely determined : which can be further evaluated using the expansion1.32). Theorem 2.1 Characterization of Conditional Objects. I’ oolanFor the formal...naturalditional forms is to determine if there i - e mapping class sense to v:’. gm, W:X . fn,VKW;; l.. +n , re-from. these higher levels down to the sin

Barcoding Human Physical Activity to Assess Chronic Pain Conditions

PubMed Central

Paraschiv-Ionescu, Anisoara; Perruchoud, Christophe; Buchser, Eric; Aminian, Kamiar

2012-01-01

Background Modern theories define chronic pain as a multidimensional experience – the result of complex interplay between physiological and psychological factors with significant impact on patients' physical, emotional and social functioning. The development of reliable assessment tools capable of capturing the multidimensional impact of chronic pain has challenged the medical community for decades. A number of validated tools are currently used in clinical practice however they all rely on self-reporting and are therefore inherently subjective. In this study we show that a comprehensive analysis of physical activity (PA) under real life conditions may capture behavioral aspects that may reflect physical and emotional functioning. Methodology PA was monitored during five consecutive days in 60 chronic pain patients and 15 pain-free healthy subjects. To analyze the various aspects of pain-related activity behaviors we defined the concept of PA ‘barcoding’. The main idea was to combine different features of PA (type, intensity, duration) to define various PA states. The temporal sequence of different states was visualized as a ‘barcode’ which indicated that significant information about daily activity can be contained in the amount and variety of PA states, and in the temporal structure of sequence. This information was quantified using complementary measures such as structural complexity metrics (information and sample entropy, Lempel-Ziv complexity), time spent in PA states, and two composite scores, which integrate all measures. The reliability of these measures to characterize chronic pain conditions was assessed by comparing groups of subjects with clinically different pain intensity. Conclusion The defined measures of PA showed good discriminative features. The results suggest that significant information about pain-related functional limitations is captured by the structural complexity of PA barcodes, which decreases when the intensity of pain

Pre-Service Teachers' Free and Structured Mathematical Problem Posing

ERIC Educational Resources Information Center

Silber, Steven; Cai, Jinfa

2017-01-01

This exploratory study examined how pre-service teachers (PSTs) pose mathematical problems for free and structured mathematical problem-posing conditions. It was hypothesized that PSTs would pose more complex mathematical problems under structured posing conditions, with increasing levels of complexity, than PSTs would pose under free posing…

Stem cell therapies for age-related macular degeneration: the past, present, and future.

PubMed

Dang, Yalong; Zhang, Chun; Zhu, Yu

2015-01-01

In the developed world, age-related macular degeneration (AMD) is one of the major causes of irreversible blindness in the elderly. Although management of neovascular AMD (wet AMD) has dramatically progressed, there is still no effective treatment for nonneovascular AMD (dry AMD), which is characterized by retinal pigment epithelial (RPE) cell death (or dysfunction) and microenvironmental disruption in the retina. Therefore, RPE replacement and microenvironmental regulation represent viable treatments for dry AMD. Recent advances in cell biology have demonstrated that RPE cells can be easily generated from several cell types (pluripotent stem cells, multipotent stem cells, or even somatic cells) by spontaneous differentiation, coculturing, defined factors or cell reprogramming, respectively. Additionally, in vivo studies also showed that the restoration of visual function could be obtained by transplanting functional RPE cells into the subretinal space of recipient. More importantly, clinical trials approved by the US government have shown promising prospects in RPE transplantation. However, key issues such as implantation techniques, immune rejection, and xeno-free techniques are still needed to be further investigated. This review will summarize recent advances in cell transplantation for dry AMD. The obstacles and prospects in this field will also be discussed.

Human Adipose-Derived Mesenchymal Stem/Stromal Cells Handling Protocols. Lipid Droplets and Proteins Double-Staining

PubMed Central

Gojanovich, Aldana D.; Gimenez, María C.; Masone, Diego; Rodriguez, Tania M.; Dewey, Ricardo A.; Delgui, Laura R.; Bustos, Diego M.; Uhart, Marina

2018-01-01

Human Adipose-derived mesenchymal stem/stromal cells (hASCs) are of great interest because of their potential for therapeutic approaches. The method described here covers every single step necessary for hASCs isolation from subcutaneous abdominal adipose tissue, multicolor phenotyping by flow cytometry, and quantitative determination of adipogenic differentiation status by means of lipid droplets (LDs) accumulation, and Western blot analysis. Moreover, to simultaneously analyze both LDs accumulation and cellular proteins localization by fluorescence microscopy, we combined Oil Red O (ORO) staining with immunofluorescence detection. For LDs quantification we wrote a program for automatic ORO-stained digital image processing implemented in Octave, a freely available software package. Our method is based on the use of the traditional low cost neutral lipids dye ORO, which can be imaged both by bright-field and fluorescence microscopy. The utilization of ORO instead of other more expensive lipid-specific dyes, together with the fact that the whole method has been designed employing cost-effective culture reagents (standard culture medium and serum), makes it affordable for tight-budget research laboratories. These may be replaced, if necessary or desired, by defined xeno-free reagents for clinical research and applications. PMID:29670879

Human Adipose-Derived Mesenchymal Stem/Stromal Cells Handling Protocols. Lipid Droplets and Proteins Double-Staining.

PubMed

Gojanovich, Aldana D; Gimenez, María C; Masone, Diego; Rodriguez, Tania M; Dewey, Ricardo A; Delgui, Laura R; Bustos, Diego M; Uhart, Marina

2018-01-01

Human Adipose-derived mesenchymal stem/stromal cells (hASCs) are of great interest because of their potential for therapeutic approaches. The method described here covers every single step necessary for hASCs isolation from subcutaneous abdominal adipose tissue, multicolor phenotyping by flow cytometry, and quantitative determination of adipogenic differentiation status by means of lipid droplets (LDs) accumulation, and Western blot analysis. Moreover, to simultaneously analyze both LDs accumulation and cellular proteins localization by fluorescence microscopy, we combined Oil Red O (ORO) staining with immunofluorescence detection. For LDs quantification we wrote a program for automatic ORO-stained digital image processing implemented in Octave, a freely available software package. Our method is based on the use of the traditional low cost neutral lipids dye ORO, which can be imaged both by bright-field and fluorescence microscopy. The utilization of ORO instead of other more expensive lipid-specific dyes, together with the fact that the whole method has been designed employing cost-effective culture reagents (standard culture medium and serum), makes it affordable for tight-budget research laboratories. These may be replaced, if necessary or desired, by defined xeno-free reagents for clinical research and applications.

Scalable topographies to support proliferation and Oct4 expression by human induced pluripotent stem cells

PubMed Central

Reimer, Andreas; Vasilevich, Aliaksei; Hulshof, Frits; Viswanathan, Priyalakshmi; van Blitterswijk, Clemens A.; de Boer, Jan; Watt, Fiona M.

2016-01-01

It is well established that topographical features modulate cell behaviour, including cell morphology, proliferation and differentiation. To define the effects of topography on human induced pluripotent stem cells (iPSC), we plated cells on a topographical library containing over 1000 different features in medium lacking animal products (xeno-free). Using high content imaging, we determined the effect of each topography on cell proliferation and expression of the pluripotency marker Oct4 24 h after seeding. Features that maintained Oct4 expression also supported proliferation and cell-cell adhesion at 24 h, and by 4 days colonies of Oct4-positive, Sox2-positive cells had formed. Computational analysis revealed that small feature size was the most important determinant of pluripotency, followed by high wave number and high feature density. Using this information we correctly predicted whether any given topography within our library would support the pluripotent state at 24 h. This approach not only facilitates the design of substrates for optimal human iPSC expansion, but also, potentially, identification of topographies with other desirable characteristics, such as promoting differentiation. PMID:26757610

Comparison of wrist-worn and hip-worn activity monitors under free living conditions.

PubMed

Hargens, Trent A; Deyarmin, Kayla N; Snyder, Kelsey M; Mihalik, Allison G; Sharpe, Lauren E

2017-04-01

Current recommendations state that individuals engage in 150 min of moderate or 75 min of vigorous intensity physical activity (PA) each week. Commercial PA monitors are becoming popular for everyday use. The accuracy of these devices, however, is not well understood. We sought to examine the accuracy of two commercial devices, one wrist and one hip-worn, under free-living conditions. Twenty-two subjects wore two commercially available devices and one ActiGraph (AG) for seven consecutive days under normal activity. Mean steps per day between all three devices differed significantly. No differences were found in moderate-to-vigorous intensity physical activity (MPVA). Daily energy expenditure (EE) also differed significantly between the AG and the commercial devices. Bland-Altman analysis found poor agreement between the AG and the commercial devices with regards to steps and EE, but good agreement in MVPA. Results suggest that the commercial devices are less accurate in estimating steps and EE. These devices did show good agreement with regards to MVPA, suggesting that they may provide useful feedback for individuals seeking to achieve the current PA guidelines for MVPA. Improvements are needed with regards to steps and EE estimation.

The world of "GM-free".

PubMed

Moses, Vivian; Brookes, Graham

2013-01-01

The rapid global development of agricultural production systems using seeds derived from genetic modification (GM) has been paralleled by the growth of attempts to keep at least a part of the world's agriculture and food as free from GM-crops and their products as possible. The ideal for some proponents of such "GM-free" activity would be a total absence, usually styled "zero content"; others, perhaps more realistically, opt for a definition usually not precisely defined that allows for minimal trace levels of GM material. The reasons for wanting "GM-free" agriculture and its products are varied; they include philosophical and religious beliefs, concern for human (and animal) health--and for "the environment"-as well as commercial and political interests. With such a variety of motivations, and in the absence of legal rulings, the definitions of "GM-free" vary according to inclination and circumstances. Whatever the precise meaning, the maintenance of a "GM-free" product stream in a world where GM crop production is widespread requires the establishment of identity preservation and segregation systems in which traceability and testing are cornerstones. Inevitably these have cost implications for the supply chain and/or the ultimate consumer. In a number of countries different forms of "GM-free" labels exist for some products; the style of such labels is variable with schemes and labels typically voluntary or privately organized. In more recent years, some governments have begun to regularize the definition and meaning of "GM-free." We conclude our analysis by exploring consumer reactions both to "GM-free" and to "GM-free" labels, and ask who ultimately benefits from preserving a product stream substantially or entirely devoid of GM-content.

Multivariable extrapolation of grand canonical free energy landscapes

NASA Astrophysics Data System (ADS)

Mahynski, Nathan A.; Errington, Jeffrey R.; Shen, Vincent K.

2017-12-01

We derive an approach for extrapolating the free energy landscape of multicomponent systems in the grand canonical ensemble, obtained from flat-histogram Monte Carlo simulations, from one set of temperature and chemical potentials to another. This is accomplished by expanding the landscape in a Taylor series at each value of the order parameter which defines its macrostate phase space. The coefficients in each Taylor polynomial are known exactly from fluctuation formulas, which may be computed by measuring the appropriate moments of extensive variables that fluctuate in this ensemble. Here we derive the expressions necessary to define these coefficients up to arbitrary order. In principle, this enables a single flat-histogram simulation to provide complete thermodynamic information over a broad range of temperatures and chemical potentials. Using this, we also show how to combine a small number of simulations, each performed at different conditions, in a thermodynamically consistent fashion to accurately compute properties at arbitrary temperatures and chemical potentials. This method may significantly increase the computational efficiency of biased grand canonical Monte Carlo simulations, especially for multicomponent mixtures. Although approximate, this approach is amenable to high-throughput and data-intensive investigations where it is preferable to have a large quantity of reasonably accurate simulation data, rather than a smaller amount with a higher accuracy.

Water-tolerant and reusable Lewis acid catalyst for the one-pot synthesis of 3,4-dihydropyrimidin-2-(1H)-ones under solvent-free conditions.

PubMed

Wang, Min; Song, Zhiguo; Jiang, Heng; Gong, Hong

2010-01-01

3,4-Dihydropyrimidin-2-(1H)-ones were synthesized in high yields by a one-pot cyclocondensation of an aldehyde, a 1,3-dicarbonyl compound, and urea using copper methanesulfonate (2 mol%) as a recyclable catalyst under solvent-free conditions in short reaction time (1-2 h).

Reference-free fatigue crack detection using nonlinear ultrasonic modulation under various temperature and loading conditions

NASA Astrophysics Data System (ADS)

Lim, Hyung Jin; Sohn, Hoon; DeSimio, Martin P.; Brown, Kevin

2014-04-01

This study presents a reference-free fatigue crack detection technique using nonlinear ultrasonic modulation. When low frequency (LF) and high frequency (HF) inputs generated by two surface-mounted lead zirconate titanate (PZT) transducers are applied to a structure, the presence of a fatigue crack can provide a mechanism for nonlinear ultrasonic modulation and create spectral sidebands around the frequency of the HF signal. The crack-induced spectral sidebands are isolated using a combination of linear response subtraction (LRS), synchronous demodulation (SD) and continuous wavelet transform (CWT) filtering. Then, a sequential outlier analysis is performed on the extracted sidebands to identify the crack presence without referring any baseline data obtained from the intact condition of the structure. Finally, the robustness of the proposed technique is demonstrated using actual test data obtained from simple aluminum plate and complex aircraft fitting-lug specimens under varying temperature and loading variations.

ENVIRONMENTALLY BENIGN ORGANIC TRANSFORMATIONS USING MICROWAVE IRRADIATION UNDER SOLVENT-FREE CONDITIONS

EPA Science Inventory

Microwave-expedited solvent-free synthetic processes involve the exposure of neat reactants to microwave (MW) irradiation in the presence of supported reagents or catalysts on mineral oxides. Recent developments are described and the salient features of these high yield protocols...

Supersonic Free-Jet Combustion in a Ramjet Burner

NASA Technical Reports Server (NTRS)

Trefny, Charles J.; Dippold, Vance F., III

2010-01-01

A new dual-mode ramjet combustor concept intended for operation over a wide flight Mach number range is described. Subsonic combustion mode is similar to that of a traditional ram combustor which allows operation at higher efficiency, and to lower flight Mach numbers than current dual-mode scramjets. High speed mode is characterized by supersonic combustion in a free-jet that traverses the subsonic combustion chamber to a variable nozzle. The maximum flight Mach number of this scheme is governed largely by the same physics as its classical counterpart. Although a variable combustor exit aperture is required, the need for fuel staging to accommodate the combustion process is eliminated. Local heating from shock-boundary-layer interactions on combustor walls is also eliminated. Given the parallel nature of the present scheme, overall flowpath length is less than that of present dual-mode configurations. Cycle analysis was done to define the flowpath geometry for computational fluid dynamics (CFD) analysis, and then to determine performance based on the CFD results. CFD results for Mach 5, 8, and 12 flight conditions indicate stable supersonic free-jet formation and nozzle reattachment, thereby establishing the basic feasibility of the concept. These results also reveal the structure of, and interactions between the free-jet and recirculating combustion chamber flows. Performance based on these CFD results is slightly less than that of the constant-pressure-combustion cycle analysis primarily due to these interactions. These differences are quantified and discussed. Additional CFD results at the Mach 8 flight condition show the effects of nozzle throat area variation on combustion chamber pressure, flow structure, and performance. Calculations with constant temperature walls were also done to evaluate heat flux and overall heat loads. Aspects of the concept that warrant further study are outlined. These include diffuser design, ramjet operation, mode transition, loss

The Effects of Defined Benefit Pension Incentives and Working Conditions on Teacher Retirement Decisions

ERIC Educational Resources Information Center

Furgeson, Joshua; Strauss, Robert P.; Vogt, William B.

2006-01-01

The retirement behavior of Pennsylvania public school teachers in 1997-98 and 1998-99, a period when state early retirement incentives were temporarily increased, is modeled using a choice framework that emphasizes both pecuniary and nonpecuniary factors of the retirement decision under a defined benefit retirement plan. We find each to have large…

Error Free Software

NASA Technical Reports Server (NTRS)

1985-01-01

A mathematical theory for development of "higher order" software to catch computer mistakes resulted from a Johnson Space Center contract for Apollo spacecraft navigation. Two women who were involved in the project formed Higher Order Software, Inc. to develop and market the system of error analysis and correction. They designed software which is logically error-free, which, in one instance, was found to increase productivity by 600%. USE.IT defines its objectives using AXES -- a user can write in English and the system converts to computer languages. It is employed by several large corporations.

Bond strength durability of self-etching adhesives and resin cements to dentin.

PubMed

Chaves, Carolina de Andrade Lima; de Melo, Renata Marques; Passos, Sheila Pestana; Camargo, Fernanda Pelógia; Bottino, Marco Antonio; Balducci, Ivan

2009-01-01

To evaluate the microtensile bond strength (microTBS) of one- (Xeno III, Dentsply) and two-step (Tyrian-One Step Plus, Bisco) self-etching adhesive systems bonded to dentin and cemented to chemically cured (C&B Metabond) or light-cured paste of a dual-cure resin cement (Variolink II, Ivoclar) within a short (24 h) and long period of evaluation (90 days). Forty recently extracted human molars had their roots removed and their occlusal dentin exposed and ground wet with 600-grit SiC paper. After application of one of the adhesives, the resin cement was applied to the bonded surface and a composite resin block was incrementally built up to a height of 5 mm (n=10). The restored teeth were stored in distilled water at 37 degrees C for 7 days. The teeth were then cut along two axes (x and y), producing beam-shaped specimens with 0.8 mm(2) cross-sectional area, which were subjected to microTBS testing at a crosshead speed of 0.05 mm/min and stressed to failure after 24 h or 90 days of storage in water. The microTBS data in MPa were subjected to three-way analysis of variance and Tukey's test (alpha= 0.05). The interaction effect for all three factors was statistically significant (three-way ANOVA, p<0.001). All eight experimental means (MPa) were compared by the Tukey's test (p<0.05) and the following results were obtained: Tyrian-One Step Plus /C&B/24 h (22.4+/-7.3); Tyrian-One Step Plus /Variolink II/24 h (39.4+/-11.6); Xeno III/C&B/24 h (40.3+/-12.9); Xeno III/Variolink II/24 h (25.8+/-10.5); Tyrian-One Step Plus /C&B/90 d (22.1+/-12.8) Tyrian-One Step Plus/VariolinkII/90 d (24.2+/-14.2); Xeno III/C&B/90 d (27.0+/-13.5); Xeno III/Variolink II/90 d (33.0+/-8.9). Xeno III/Variolink II was the luting agent/adhesive combination that provided the most promising bond strength after 90 days of storage in water.

BOND STRENGTH DURABILITY OF SELF-ETCHING ADHESIVES AND RESIN CEMENTS TO DENTIN

PubMed Central

Chaves, Carolina de Andrade Lima; de Melo, Renata Marques; Passos, Sheila Pestana; Camargo, Fernanda Pelógia; Bottino, Marco Antonio; Balducci, Ivan

2009-01-01

Objectives: To evaluate the microtensile bond strength (μTBS) of one- (Xeno III, Dentsply) and two-step (Tyrian-One Step Plus, Bisco) self-etching adhesive systems bonded to dentin and cemented to chemically cured (C&B Metabond) or light-cured paste of a dual-cure resin cement (Variolink II, Ivoclar) within a short (24 h) and long period of evaluation (90 days). Material and Methods: Forty recently extracted human molars had their roots removed and their occlusal dentin exposed and ground wet with 600-grit SiC paper. After application of one of the adhesives, the resin cement was applied to the bonded surface and a composite resin block was incrementally built up to a height of 5 mm (n=10). The restored teeth were stored in distilled water at 37°C for 7 days. The teeth were then cut along two axes (x and y), producing beam-shaped specimens with 0.8 mm2 cross-sectional area, which were subjected to μTBS testing at a crosshead speed of 0.05 mm/min and stressed to failure after 24 h or 90 days of storage in water. The μTBS data in MPa were subjected to three-way analysis of variance and Tukey's test (α= 0.05). Results: The interaction effect for all three factors was statistically significant (three-way ANOVA, p<0.001). All eight experimental means (MPa) were compared by the Tukey's test (p<0.05) and the following results were obtained: Tyrian-One Step Plus/C&B/24 h (22.4±7.3); Tyrian-One Step Plus/Variolink II/24 h (39.4±11.6); Xeno III/C&B/24 h (40.3±12.9); Xeno III/Variolink II/24 h (25.8±10.5); Tyrian-One Step Plus/C&B/90 d (22.1±12.8) Tyrian-One Step Plus/VariolinkII/90 d (24.2±14.2); Xeno III/C&B/90 d (27.0±13.5); Xeno III/Variolink II/ 90 d (33.0±8.9). Conclusions: Xeno III/Variolink II was the luting agent/adhesive combination that provided the most promising bond strength after 90 days of storage in water. PMID:19466243

CHEMISTRY UNDER NON-TRADITIONAL CONDITIONS: SOLVENT-FREE SYNTHESIS USING MICROWAVE IRRADIATION

EPA Science Inventory

A solvent-free approach that involves microwave (MW) irradiation of neat reactants (undiluted) catalyzed by the surfaces of less-expensive and recyclable mineral supports such as alumina, silica, clay, or "doped" surfaces is presented which is applicable to a wide range of cleava...

Ethanol self-administration in free-flying honeybees (Apis mellifera L.) in an operant conditioning protocol.

PubMed

Sokolowski, Michel B C; Abramson, Charles I; Craig, David Philip Arthur

2012-09-01

This study examines the effect of ethanol (EtOH) on continuous reinforcement schedules in the free-flying honeybee (Apis mellifera L.). As fermented nectars may be encountered naturally in the environment, we designed an experiment combining the tools of laboratory research with minimal disturbance to the natural life of honeybees. Twenty-five honeybees were trained to fly from their colonies to a fully automated operant chamber with head poking as the operant response. Load size, intervisit interval, and interresponse times (IRTs) served as the dependent variables and were monitored over the course of a daily training session consisting of many visits. Experimental bees were tested using an ABA design in which sucrose only was administered during condition A and a 5% EtOH sucrose solution was administered during condition B. Control bees received sucrose solution only. Most bees continued to forage after EtOH introduction. EtOH significantly reduced the load size and the intervisit interval with no significant effect on IRTs. However, a look on individual data shows large individual differences suggesting the existence of different kinds of behavioral phenotypes linked to EtOH consumption and effects. Our results contribute to the study of EtOH consumption as a normal phenomenon in an ecological context and open the door to schedule-controlled drug self-administration studies in honeybees. Copyright © 2012 by the Research Society on Alcoholism.

Microwave-Assisted Condensation Reactions of Acetophenone Derivatives and Activated Methylene Compounds with Aldehydes Catalyzed by Boric Acid under Solvent-Free Conditions.

PubMed

Brun, Elodie; Safer, Abdelmounaim; Carreaux, François; Bourahla, Khadidja; L'helgoua'ch, Jean-Martial; Bazureau, Jean-Pierre; Villalgordo, Jose Manuel

2015-06-23

We here disclosed a new protocol for the condensation of acetophenone derivatives and active methylene compounds with aldehydes in the presence of boric acid under microwave conditions. Implementation of the reaction is simple, healthy and environmentally friendly owing to the use of a non-toxic catalyst coupled to a solvent-free procedure. A large variety of known or novel compounds have thus been prepared, including with substrates bearing acid or base-sensitive functional groups.

Well-posedness of the free boundary problem in compressible elastodynamics

NASA Astrophysics Data System (ADS)

Trakhinin, Yuri

2018-02-01

We study the free boundary problem for the flow of a compressible isentropic inviscid elastic fluid. At the free boundary moving with the velocity of the fluid particles the columns of the deformation gradient are tangent to the boundary and the pressure vanishes outside the flow domain. We prove the local-in-time existence of a unique smooth solution of the free boundary problem provided that among three columns of the deformation gradient there are two which are non-collinear vectors at each point of the initial free boundary. If this non-collinearity condition fails, the local-in-time existence is proved under the classical Rayleigh-Taylor sign condition satisfied at the first moment. By constructing an Hadamard-type ill-posedness example for the frozen coefficients linearized problem we show that the simultaneous failure of the non-collinearity condition and the Rayleigh-Taylor sign condition leads to Rayleigh-Taylor instability.

Syndromic autism spectrum disorders: moving from a clinically defined to a molecularly defined approach

PubMed Central

Fernandez, Bridget A.; Scherer, Stephen W.

2017-01-01

Autism spectrum disorder (ASD) encompasses a group of neurodevelopmental conditions diagnosed solely on the basis of behavioral assessments that reveal social deficits. Progress has been made in understanding its genetic underpinnings, but most ASD-associated genetic variants, which include copy number variants (CNVs) and mutations in ASD-risk genes, account for no more than 1 % of ASD cases. This high level of genetic heterogeneity leads to challenges obtaining and interpreting genetic testing in clinical settings. The traditional definition of syndromic ASD is a disorder with a clinically defined pattern of somatic abnormalities and a neurobehavioral phenotype that may include ASD. Most have a known genetic cause. Examples include fragile X syndrome and tuberous sclerosis complex. We propose dividing syndromic autism into the following two groups: (i) ASD that occurs in the context of a clinically defined syndrome-recognizing these disorders depends on the familiarity of the clinician with the features of the syndrome, and the diagnosis is typically confirmed by targeted genetic testing (eg, mutation screening of FMR1); (ii) ASD that occurs as a feature of a molecularly defined syndrome-for this group of patients, ASD-associated variants are identified by genome-wide testing that is not hypothesis driven (eg, microarray, whole exome sequencing). These ASD groups cannot be easily clinically defined because patients with a given variant have variable somatic abnormalities (dysmorphism and birth defects). In this article, we review common diagnoses from the above categories and suggest a testing strategy for patients, guided by determining whether the individual has essential or complex ASD; patients in the latter group have multiple morphologic anomalies on physical examination. Finally, we recommend that the syndromic versus nonsyndromic designation ultimately be replaced by classification of ASD according to its genetic etiology, which will inform about the

Culture conditions defining glioblastoma cells behavior: what is the impact for novel discoveries?

PubMed

Ledur, Pítia Flores; Onzi, Giovana Ravizzoni; Zong, Hui; Lenz, Guido

2017-09-15

In cancer research, the use of established cell lines has gradually been replaced by primary cell cultures due to their better representation of in vivo cancer cell behaviors. However, a major challenge with primary culture involves the finding of growth conditions that minimize alterations in the biological state of the cells. To ensure reproducibility and translational potentials for research findings, culture conditions need to be chosen so that the cell population in culture best mimics tumor cells in vivo . Glioblastoma (GBM) is one of the most aggressive and heterogeneous tumor types and the GBM research field would certainly benefit from culture conditions that could maintain the original plethora of phenotype of the cells. Here, we review culture media and supplementation options for GBM cultures, the rationale behind their use, and how much those choices affect drug-screening outcomes. We provide an overview of 120 papers that use primary GBM cultures and discuss the current predominant conditions. We also show important primary research data indicating that "mis-cultured" glioma cells can acquire unnatural drug sensitivity, which would have devastating effects for clinical translations. Finally, we propose the concurrent test of four culture conditions to minimize the loss of cell coverage in culture.

Two-year Randomized Clinical Trial of Self-etching Adhesives and Selective Enamel Etching.

PubMed

Pena, C E; Rodrigues, J A; Ely, C; Giannini, M; Reis, A F

2016-01-01

The aim of this randomized, controlled prospective clinical trial was to evaluate the clinical effectiveness of restoring noncarious cervical lesions with two self-etching adhesive systems applied with or without selective enamel etching. A one-step self-etching adhesive (Xeno V(+)) and a two-step self-etching system (Clearfil SE Bond) were used. The effectiveness of phosphoric acid selective etching of enamel margins was also evaluated. Fifty-six cavities were restored with each adhesive system and divided into two subgroups (n=28; etch and non-etch). All 112 cavities were restored with the nanohybrid composite Esthet.X HD. The clinical effectiveness of restorations was recorded in terms of retention, marginal integrity, marginal staining, caries recurrence, and postoperative sensitivity after 3, 6, 12, 18, and 24 months (modified United States Public Health Service). The Friedman test detected significant differences only after 18 months for marginal staining in the groups Clearfil SE non-etch (p=0.009) and Xeno V(+) etch (p=0.004). One restoration was lost during the trial (Xeno V(+) etch; p>0.05). Although an increase in marginal staining was recorded for groups Clearfil SE non-etch and Xeno V(+) etch, the clinical effectiveness of restorations was considered acceptable for the single-step and two-step self-etching systems with or without selective enamel etching in this 24-month clinical trial.

Snacks containing whey protein and polydextrose induce a sustained reduction in daily energy intake over 2 wk under free-living conditions.

PubMed

Astbury, Nerys M; Taylor, Moira A; French, Stephen J; Macdonald, Ian A

2014-05-01

The manipulation of the composition of foods consumed as between-meal snacks may aid daily energy restriction. We compared the effects of the consumption of 2 energy-matched snack bars on appetite, energy intake (EI), and metabolic and endocrine responses. In addition, we investigated whether the acute effects of the consumption of snacks were maintained under free-living conditions and whether the habitual daily consumption of the snack over 14 d influenced these effects. Ten lean men [mean ± SD age: 30.7 ± 9.7 y; body mass index (in kg/m(2)): 23.2 ± 2.8] consumed a whey protein and polydextrose (PPX) snack bar or an isoenergetic control snack bar as a midmorning, between-meal snack for 14 consecutive days in a double-blind, randomized, crossover design. The two 14-d intervention phases were separated by a 14-d washout period. On the first (day 1) and last (day 15) days of each intervention phase, appetite, food intake, and blood metabolite and endocrine responses were assessed under laboratory conditions. Free-living EI was recorded on days 4, 8, and 12 of interventions. Total daily EI was significantly lower when the PPX snack was consumed during experimental days (10,149 ± 831 compared with 11,931 ± 896 kJ; P < 0.01), and daily EI remained lower when the PPX snack was consumed during the free-living part of the intervention (7904 ± 610 compared with 9041 ± 928 kJ; P < 0.05). The PPX snack was associated with lower glucose and ghrelin and higher glucagon-like peptide 1 and peptide tyrosine-tyrosine responses. The manipulation of the composition of foods consumed as snacks is an effective way to limit subsequent EI. This trial was registered at clinicaltrials.gov as NCT01927926.

Radiocarbon ages of terrestrial gastropods extend duration of ice-free conditions at the Two Creeks forest bed, Wisconsin, USA

USGS Publications Warehouse

Rech, Jason A.; Nekola, Jeffrey C.; Pigati, Jeffrey S.

2012-01-01

Analysis of terrestrial gastropods that underlie the late Pleistocene Two Creeks forest bed (~ 13,800–13,500 cal yr BP) in eastern Wisconsin, USA provides evidence for a mixed tundra-taiga environment prior to formation of the taiga forest bed. Ten new AMS 14C analyses on terrestrial gastropod shells indicate the mixed tundra-taiga environment persisted from ~ 14,500 to 13,900 cal yr BP. The Twocreekan climatic substage, representing ice-free conditions on the shore of Lake Michigan, therefore began near the onset of peak warming conditions during the Bølling–Allerød interstadial and lasted ~ 1000 yr, nearly 600 yr longer than previously thought. These results provide important data for understanding the response of continental ice sheets to global climate forcing and demonstrate the potential of using terrestrial gastropod fossils for both environmental reconstruction and age control in late Quaternary sediments.

Approaches to defining reference regimes for river restoration planning

NASA Astrophysics Data System (ADS)

Beechie, T. J.

2014-12-01

Reference conditions or reference regimes can be defined using three general approaches, historical analysis, contemporary reference sites, and theoretical or empirical models. For large features (e.g., floodplain channels and ponds) historical data and maps are generally reliable. For smaller features (e.g., pools and riffles in small tributaries), field data from contemporary reference sites are a reasonable surrogate for historical data. Models are generally used for features that have no historical information or present day reference sites (e.g., beaver pond habitat). Each of these approaches contributes to a watershed-wide understanding of current biophysical conditions relative to potential conditions, which helps create not only a guiding vision for restoration, but also helps quantify and locate the largest or most important restoration opportunities. Common uses of geomorphic and biological reference conditions include identifying key areas for habitat protection or restoration, and informing the choice of restoration targets. Examples of use of each of these three approaches to define reference regimes in western USA illustrate how historical information and current research highlight key restoration opportunities, focus restoration effort in areas that can produce the largest ecological benefit, and contribute to estimating restoration potential and assessing likelihood of achieving restoration goals.

D’Andrea’s disease (angiomegaly): a currently well-defined nosological entitys.

PubMed

Taurone, S; Spoletini, M; Di Matteo, F M; Mele, R; Tromba, L; Grippaudo, F R; Minni, A; Artico, M

2017-01-01

In 1997 D’Andrea et al. described a new nosological entity the characteristics of which consisted of lengthening, dilation and tortuosity of blood vessels, arteries or veins, less prominent, but also less circumscribed than an aneurysm. This condition does not necessarily imply specific aneurysm formation although aneurysms at multiple sites are a frequent observation. The term used by authors for angiomegaly of the venous system was venomegaly and the analogous condition of the arterial system was termed arteriomegaly. Although tortuosity and dilation of arteries and veins have been widely reported, suggesting a systemic disorder which affects the structural integrity of all vessels, most papers dealing with this intriguing condition did not describe any alterations in the components of vessel walls. In the present paper, the authors describe a well-defined condition, D’Andrea’s Disease (or DD, in this article), analyzing its salient morphological and clinical features and clarifying this pathological condition as a distinct and now well-defined nosological entity.

Mild intraoperative hypothermia reduces free tissue transfer thrombosis.

PubMed

Liu, Yuen-Jong; Hirsch, Brandon P; Shah, Asad A; Reid, Marjorie A; Thomson, J Grant

2011-02-01

Patients undergoing free tissue transfer are particularly susceptible to hypothermia. The goal was to investigate the impact of intraoperative core body temperature on free flap thrombosis. Two hundred twelve cases of free flap reconstruction at Yale-New Haven Hospital between 1992 and 2008 were reviewed. Free flap thrombosis was defined by complete flap necrosis or direct visualization of arterial or venous thrombosis. Temperature measurements were calibrated to bladder temperatures as measured by Foley catheter sensor. Through logistic regression analysis, maximum and minimum intraoperative temperatures were determined to be statistically significant predictors of free flap thrombosis. The optimal temperature was calculated to be 36.2 °C, and maximum intraoperative temperatures between 36.0 °C and 36.4 °C showed lower thrombosis rates than super-warmed patients ( P < 0.03). Therefore, free flap patients should be mildly hypothermic at 36.0 °C to 36.4 °C, compared with normothermia at 37.5 °C, as measured in the bladder. A prospective randomized trial investigating thrombosis rates and intraoperative temperature should be undertaken. © Thieme Medical Publishers.

Democracy and Free Speech: A Normative Theory of Society and Government.

ERIC Educational Resources Information Center

Hodge, John L.

Noting that the nature and value of free speech depends on the norms and goals of a society and on that society's appropriate form of government, this paper presents a normative theory of democracy and from that theory formulates a theory of free speech. The first section defines a number of terms used in the paper and clarifies several of the…

Tailoring rice flour structure by rubbery milling for improved gluten-free baked goods.

PubMed

Brütsch, Linda; Tribolet, Liliane; Isabettini, Stéphane; Soltermann, Patrick; Baumann, Andreas; Windhab, Erich J

2018-05-10

Ever-growing demand for gluten-free products calls for the development of novel food processing techniques to widen the range of existing baked goods. Extensive research has been targeted towards recipe optimization, widely neglecting the tailoring potential of process-induced structuring of gluten-free raw materials. Herein, we address this shortcoming by demonstrating the potential of rubbery milling for the generation of structure and techno-functionality in breads obtained from a variety of rice flour types. Moisture and temperature induced state transitions during milling were exploited to tailor the physicochemical properties of the flour. Moisture addition during conditioning of the different rice varieties and milling in the rubbery state considerably decreased starch damage due to more gentle disintegration. The degree of starch damage dictated the water absorption capacity of the rice flour types. Flour types with reduced starch damage upon milling offered lower dough densities, yielding bread loafs with a higher volume and better appearance. The choice of rice variety enables fine-tuning of the final product quality by influencing the dough viscoelasticity, which defines the final loaf volume. Whole grain rice flour dramatically increased the loaf volume, whilst simultaneously offering nutritional benefits. Combining the proposed functionalised flour types with current and future advances in product recipes paves the way towards optimised gluten-free goods.

Gravity with free initial conditions: A solution to the cosmological constant problem testable by CMB B -mode polarization

NASA Astrophysics Data System (ADS)

Totani, Tomonori

2017-10-01

In standard general relativity the universe cannot be started with arbitrary initial conditions, because four of the ten components of the Einstein's field equations (EFE) are constraints on initial conditions. In the previous work it was proposed to extend the gravity theory to allow free initial conditions, with a motivation to solve the cosmological constant problem. This was done by setting four constraints on metric variations in the action principle, which is reasonable because the gravity's physical degrees of freedom are at most six. However, there are two problems about this theory; the three constraints in addition to the unimodular condition were introduced without clear physical meanings, and the flat Minkowski spacetime is unstable against perturbations. Here a new set of gravitational field equations is derived by replacing the three constraints with new ones requiring that geodesic paths remain geodesic against metric variations. The instability problem is then naturally solved. Implications for the cosmological constant Λ are unchanged; the theory converges into EFE with nonzero Λ by inflation, but Λ varies on scales much larger than the present Hubble horizon. Then galaxies are formed only in small Λ regions, and the cosmological constant problem is solved by the anthropic argument. Because of the increased degrees of freedom in metric dynamics, the theory predicts new non-oscillatory modes of metric anisotropy generated by quantum fluctuation during inflation, and CMB B -mode polarization would be observed differently from the standard predictions by general relativity.

Effects of four-month handbike training under free-living conditions on physical fitness and health in wheelchair users.

PubMed

Hoekstra, Sven; Valent, Linda; Gobets, David; van der Woude, Lucas; de Groot, Sonja

2017-08-01

Recognizing the encouraging effect of challenging events, the HandbikeBattle (HBB) was created to promote exercise among wheelchair users. The purpose of this study was to reveal the effects on physical fitness and health outcomes of four-month handbike training under free-living conditions in preparation for the event. In this prospective cohort study, 59 relatively inexperienced handyclists participated in the HBB of 2013 or 2014. Incremental exercise tests were conducted, respiratory function was tested and anthropometrics were measured before and after the preparation period. Main outcome measures were peak power output (POpeak), peak oxygen uptake (VO2peak) and waist circumference, of which the changes were tested using repeated measures ANOVA. To detect possible determinants of changes in physical fitness, a linear regression analysis was conducted with personal characteristics, executed training volume and upper-extremity complaints during the training period as independent variables. POpeak, VO2peak and waist circumference improved significantly with 17%, 7% and 4.1%, respectively. None of the included variables were significant determinants for the changes in POpeak found as a result of the training. A challenging event such as the HBB provokes training regimes among participants of sufficient load to realize substantial improvements in physical fitness and health outcomes. Implications for Rehabilitation Due to the often impaired muscle function in the lower-limbs and an inactive lifestyle, wheelchair users generally show considerably lower levels of fitness compared to able-bodied individuals. This prospective cohort study showed that four months of handbike training under free-living conditions in preparation for this event resulted in substantial improvements in physical fitness and health outcomes in wheelchair users. The creation of a challenging event such as the HandbikeBattle as part of a follow-up rehabilitation practice can therefore be a useful

Comparing Free-Free and Shaker Table Model Correlation Methods Using Jim Beam

NASA Technical Reports Server (NTRS)

Ristow, James; Smith, Kenneth Wayne, Jr.; Johnson, Nathaniel; Kinney, Jackson

2018-01-01

Finite element model correlation as part of a spacecraft program has always been a challenge. For any NASA mission, the coupled system response of the spacecraft and launch vehicle can be determined analytically through a Coupled Loads Analysis (CLA), as it is not possible to test the spacecraft and launch vehicle coupled system before launch. The value of the CLA is highly dependent on the accuracy of the frequencies and mode shapes extracted from the spacecraft model. NASA standards require the spacecraft model used in the final Verification Loads Cycle to be correlated by either a modal test or by comparison of the model with Frequency Response Functions (FRFs) obtained during the environmental qualification test. Due to budgetary and time constraints, most programs opt to correlate the spacecraft dynamic model during the environmental qualification test, conducted on a large shaker table. For any model correlation effort, the key has always been finding a proper definition of the boundary conditions. This paper is a correlation case study to investigate the difference in responses of a simple structure using a free-free boundary, a fixed boundary on the shaker table, and a base-drive vibration test, all using identical instrumentation. The NAVCON Jim Beam test structure, featured in the IMAC round robin modal test of 2009, was selected as a simple, well recognized and well characterized structure to conduct this investigation. First, a free-free impact modal test of the Jim Beam was done as an experimental control. Second, the Jim Beam was mounted to a large 20,000 lbf shaker, and an impact modal test in this fixed configuration was conducted. Lastly, a vibration test of the Jim Beam was conducted on the shaker table. The free-free impact test, the fixed impact test, and the base-drive test were used to assess the effect of the shaker modes, evaluate the validity of fixed-base modeling assumptions, and compare final model correlation results between these

Comparison of Aperture Averaging and Receiver Diversity Techniques for Free Space Optical Links in Presence of Turbulence and Various Weather Conditions

NASA Astrophysics Data System (ADS)

Kaur, Prabhmandeep; Jain, Virander Kumar; Kar, Subrat

2014-12-01

In this paper, we investigate the performance of a Free Space Optic (FSO) link considering the impairments caused by the presence of various weather conditions such as very clear air, drizzle, haze, fog, etc., and turbulence in the atmosphere. Analytic expression for the outage probability is derived using the gamma-gamma distribution for turbulence and accounting the effect of weather conditions using the Beer-Lambert's law. The effect of receiver diversity schemes using aperture averaging and array receivers on the outage probability is studied and compared. As the aperture diameter is increased, the outage probability decreases irrespective of the turbulence strength (weak, moderate and strong) and weather conditions. Similar effects are observed when the number of direct detection receivers in the array are increased. However, it is seen that as the desired level of performance in terms of the outage probability decreases, array receiver becomes the preferred choice as compared to the receiver with aperture averaging.

SBA-15-functionalized 3-oxo-ABNO as recyclable catalyst for aerobic oxidation of alcohols under metal-free conditions.

PubMed

Karimi, Babak; Farhangi, Elham; Vali, Hojatollah; Vahdati, Saleh

2014-09-01

The nitroxyl radical 3-oxo-9-azabicyclo [3.3.1]nonane-N-oxyl (3-oxo-ABNO) has been prepared using a simple protocol. This organocatalyst is found to be an efficient catalyst for the aerobic oxidation of a wide variety of alcohols under metal-free conditions. In addition, the preparation and characterization of a supported version of 3-oxo-ABNO on ordered mesoporous silica SBA-15 (SABNO) is described for the first time. The catalyst has been characterized using several techniques including simultaneous thermal analysis (STA), transmission electron microscopy (TEM), and nitrogen sorption analysis. This catalyst exhibits catalytic performance comparable to its homogeneous analogue and much superior catalytic activity in comparison with (2,2,6,6-tetramethylpiperidin-1-yl)oxy (TEMPO) for the aerobic oxidation of almost the same range of alcohols under identical reaction conditions. It is also found that SABNO can be conveniently recovered and reused at least 12 times without significant effect on its catalytic efficiency. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

New Efficient Synthesis of 3,4-Dihydropyrimidin-2(1H)-ones Catalyzed by Benzotriazolium-Based Ionic Liquids under Solvent-Free Conditions.

PubMed

Liu, Zhiqing; Ma, Rong; Cao, Dawei; Liu, Chenjiang

2016-04-07

An efficient synthesis of novel 3,4-dihydropyrimidin-2(1H)-ones (DHPMs) and their derivatives, using Brønsted acidic ionic liquid [C₂O₂BBTA][TFA] as a catalyst, from the condensation of aryl aldehyde, β-ketoester and urea was described. Reactions proceeded smoothly for 40 min under solvent-free conditions and gave the desirable products with good to excellent yields (up to 99%). The catalyst could be easily recycled and reused with similar efficacies for at least six cycles.

Exact solution for a two-phase Stefan problem with variable latent heat and a convective boundary condition at the fixed face

NASA Astrophysics Data System (ADS)

Bollati, Julieta; Tarzia, Domingo A.

2018-04-01

Recently, in Tarzia (Thermal Sci 21A:1-11, 2017) for the classical two-phase Lamé-Clapeyron-Stefan problem an equivalence between the temperature and convective boundary conditions at the fixed face under a certain restriction was obtained. Motivated by this article we study the two-phase Stefan problem for a semi-infinite material with a latent heat defined as a power function of the position and a convective boundary condition at the fixed face. An exact solution is constructed using Kummer functions in case that an inequality for the convective transfer coefficient is satisfied generalizing recent works for the corresponding one-phase free boundary problem. We also consider the limit to our problem when that coefficient goes to infinity obtaining a new free boundary problem, which has been recently studied in Zhou et al. (J Eng Math 2017. https://doi.org/10.1007/s10665-017-9921-y).

Lead-Free Experiment in a Space Environment

NASA Technical Reports Server (NTRS)

Blanche, J. F.; Strickland, S. M.

2012-01-01

This Technical Memorandum addresses the Lead-Free Technology Experiment in Space Environment that flew as part of the seventh Materials International Space Station Experiment outside the International Space Station for approximately 18 months. Its intent was to provide data on the performance of lead-free electronics in an actual space environment. Its postflight condition is compared to the preflight condition as well as to the condition of an identical package operating in parallel in the laboratory. Some tin whisker growth was seen on a flight board but the whiskers were few and short. There were no solder joint failures, no tin pest formation, and no significant intermetallic compound formation or growth on either the flight or ground units.

Determination of the rate of crystal growth from the gas phase under conditions of turbulent free convection

NASA Astrophysics Data System (ADS)

Alad'Ev, S. I.

1987-04-01

Crystal growth in vertical and horizontal cylindrical vials, with the substrate and the source serving as the vial ends, is investigated analytically, assuming that the medium consists of a binary mixture of an active and an inert gas. The active gas is made up of the gaseous products of reactions taking place at the substrate and at the source. It is shown that turbulent free convection leads to an increase in crystal growth rate. All other conditions being equal, crystal growth in vertical vials is greater than that in horizontal ones; in both cases crystal growth rate increases with the vial radius, temperature gradient in the gas phase, and gas phase density. The results are compared with experimental data on the growth of Ge crystals in the Ge-GeI4 system.

THE FREE-FALL TIME OF FINITE SHEETS AND FILAMENTS

DOE Office of Scientific and Technical Information (OSTI.GOV)

Toala, Jesus A.; Vazquez-Semadeni, Enrique; Gomez, Gilberto C.

2012-01-10

Molecular clouds often exhibit filamentary or sheet-like shapes. We compute the free-fall time ({tau}{sub ff}) for finite, uniform, self-gravitating circular sheets and filamentary clouds of small but finite thickness, so that their volume density {rho} can still be defined. We find that, for thin sheets, the free-fall time is larger than that of a uniform sphere with the same volume density by a factor proportional to {radical}A, where the aspect ratio A is given by A = R/h, R being the sheet's radius and h is its thickness. For filamentary clouds, the aspect ratio is defined as A=L/R, where Lmore » is the filament's half-length and R is its (small) radius, and the modification factor is more complicated, although in the limit of large A it again reduces to nearly {radical}A. We propose that our result for filamentary shapes naturally explains the ubiquitous configuration of clumps fed by filaments observed in the densest structures of molecular clouds. Also, the longer free-fall times for non-spherical geometries in general may contribute toward partially alleviating the 'star formation conundrum', namely, the star formation rate in the Galaxy appears to be proceeding in a timescale much larger than the total molecular mass in the Galaxy divided by its typical free-fall time. If molecular clouds are in general formed by thin sheets and long filaments, then their relevant free-fall time may have been systematically underestimated, possibly by factors of up to one order of magnitude.« less

Impact of uncertainties in free stream conditions on the aerodynamics of a rectangular cylinder

NASA Astrophysics Data System (ADS)

Mariotti, Alessandro; Shoeibi Omrani, Pejman; Witteveen, Jeroen; Salvetti, Maria Vittoria

2015-11-01

The BARC benchmark deals with the flow around a rectangular cylinder with chord-to-depth ratio equal to 5. This flow configuration is of practical interest for civil and industrial structures and it is characterized by massively separated flow and unsteadiness. In a recent review of BARC results, significant dispersion was observed both in experimental and numerical predictions of some flow quantities, which are extremely sensitive to various uncertainties, which may be present in experiments and simulations. Besides modeling and numerical errors, in simulations it is difficult to exactly reproduce the experimental conditions due to uncertainties in the set-up parameters, which sometimes cannot be exactly controlled or characterized. Probabilistic methods and URANS simulations are used to investigate the impact of the uncertainties in the following set-up parameters: the angle of incidence, the free stream longitudinal turbulence intensity and length scale. Stochastic collocation is employed to perform the probabilistic propagation of the uncertainty. The discretization and modeling errors are estimated by repeating the same analysis for different grids and turbulence models. The results obtained for different assumed PDF of the set-up parameters are also compared.

[A free market is unfavourable for health care].

PubMed

Kant, A

2006-05-06

The new health care system in the Netherlands introduces a free market into the health care sector. Because health care lacks some essential conditions, a free market will not provide effective resource allocation in this sector. In addition, a free market leads to more bureaucracy and less solidarity, cooperation and freedom of choice. It also impairs the autonomy of the medical professional.

Deficient motion-defined and texture-defined figure-ground segregation in amblyopic children.

PubMed

Wang, Jane; Ho, Cindy S; Giaschi, Deborah E

2007-01-01

Motion-defined form deficits in the fellow eye and the amblyopic eye of children with amblyopia implicate possible direction-selective motion processing or static figure-ground segregation deficits. Deficient motion-defined form perception in the fellow eye of amblyopic children may not be fully accounted for by a general motion processing deficit. This study investigates the contribution of figure-ground segregation deficits to the motion-defined form perception deficits in amblyopia. Performances of 6 amblyopic children (5 anisometropic, 1 anisostrabismic) and 32 control children with normal vision were assessed on motion-defined form, texture-defined form, and global motion tasks. Performance on motion-defined and texture-defined form tasks was significantly worse in amblyopic children than in control children. Performance on global motion tasks was not significantly different between the 2 groups. Faulty figure-ground segregation mechanisms are likely responsible for the observed motion-defined form perception deficits in amblyopia.

Conditioning and Robustness of RNA Boltzmann Sampling under Thermodynamic Parameter Perturbations.

PubMed

Rogers, Emily; Murrugarra, David; Heitsch, Christine

2017-07-25

Understanding how RNA secondary structure prediction methods depend on the underlying nearest-neighbor thermodynamic model remains a fundamental challenge in the field. Minimum free energy (MFE) predictions are known to be "ill conditioned" in that small changes to the thermodynamic model can result in significantly different optimal structures. Hence, the best practice is now to sample from the Boltzmann distribution, which generates a set of suboptimal structures. Although the structural signal of this Boltzmann sample is known to be robust to stochastic noise, the conditioning and robustness under thermodynamic perturbations have yet to be addressed. We present here a mathematically rigorous model for conditioning inspired by numerical analysis, and also a biologically inspired definition for robustness under thermodynamic perturbation. We demonstrate the strong correlation between conditioning and robustness and use its tight relationship to define quantitative thresholds for well versus ill conditioning. These resulting thresholds demonstrate that the majority of the sequences are at least sample robust, which verifies the assumption of sampling's improved conditioning over the MFE prediction. Furthermore, because we find no correlation between conditioning and MFE accuracy, the presence of both well- and ill-conditioned sequences indicates the continued need for both thermodynamic model refinements and alternate RNA structure prediction methods beyond the physics-based ones. Copyright © 2017. Published by Elsevier Inc.

Video-Recorded Validation of Wearable Step Counters under Free-living Conditions.

PubMed

Toth, Lindsay P; Park, Susan; Springer, Cary M; Feyerabend, McKenzie D; Steeves, Jeremy A; Bassett, David R

2018-06-01

The purpose of this study was to determine the accuracy of 14-step counting methods under free-living conditions. Twelve adults (mean ± SD age, 35 ± 13 yr) wore a chest harness that held a GoPro camera pointed down at the feet during all waking hours for 1 d. The GoPro continuously recorded video of all steps taken throughout the day. Simultaneously, participants wore two StepWatch (SW) devices on each ankle (all programmed with different settings), one activPAL on each thigh, four devices at the waist (Fitbit Zip, Yamax Digi-Walker SW-200, New Lifestyles NL-2000, and ActiGraph GT9X (AG)), and two devices on the dominant and nondominant wrists (Fitbit Charge and AG). The GoPro videos were downloaded to a computer and researchers counted steps using a hand tally device, which served as the criterion method. The SW devices recorded between 95.3% and 102.8% of actual steps taken throughout the day (P > 0.05). Eleven step counting methods estimated less than 100% of actual steps; Fitbit Zip, Yamax Digi-Walker SW-200, and AG with the moving average vector magnitude algorithm on both wrists recorded 71% to 91% of steps (P > 0.05), whereas the activPAL, New Lifestyles NL-2000, and AG (without low-frequency extension (no-LFE), moving average vector magnitude) worn on the hip, and Fitbit Charge recorded 69% to 84% of steps (P < 0.05). Five methods estimated more than 100% of actual steps; AG (no-LFE) on both wrists recorded 109% to 122% of steps (P > 0.05), whereas the AG (LFE) on both wrists and the hip recorded 128% to 220% of steps (P < 0.05). Across all waking hours of 1 d, step counts differ between devices. The SW, regardless of settings, was the most accurate method of counting steps.

Catalyst-free ethyl biodiesel production from rice bran under subcritical condition

NASA Astrophysics Data System (ADS)

Zullaikah, Siti; Afifudin, Riza; Amalia, Rizky

2015-12-01

In-situ ethyl biodiesel production from rice bran under subcritical water and ethanol with no catalyst was employed. This process is environmentally friendly and is very flexible in term of feedstock utilization since it can handle relatively high moisture and free fatty acids (FFAs) contents. In addition, the alcohol, i.e. bioethanol, is a non-toxic, biodegradable, and green raw material when produced from non-edible biomass residues, leading to a 100% renewable biodiesel. The fatty acid ethyl esters (FAEEs, ethyl biodiesel) are better than fatty acid methyl esters (FAMEs, methyl biodiesel) in terms of fuel properties, including cetane number, oxidation stability and cold flow properties. The influences of the operating variables such as reaction time (1 - 10 h), ethanol concentration (12.5 - 87.5%), and pressurizing gas (N2 and CO2) on the ethyl biodiesel yield and purity have been investigated systematically while the temperature and pressure were kept constant at 200 °C and 40 bar. The optimum results were obtained at 5 h reaction time and 75% ethanol concentration using CO2 as compressing gas. Ethyl biodiesel yield and purity of 58.78% and 61.35%, respectively, were obtained using rice bran with initial FFAs content of 37.64%. FFAs level was reduced to 14.22% with crude ethyl biodiesel recovery of 95.98%. Increasing the reaction time up to 10 h only increased the yield and purity by only about 3%. Under N2 atmosphere and at the same operating conditions (5h and 75% ethanol), ethyl biodiesel yield and purity decreased to 54.63% and 58.07%, respectively, while FFAs level was increased to 17.93% and crude ethyl biodiesel recovery decreased to 87.32%.

Free response approach in a parametric system

NASA Astrophysics Data System (ADS)

Huang, Dishan; Zhang, Yueyue; Shao, Hexi

2017-07-01

In this study, a new approach to predict the free response in a parametric system is investigated. It is proposed in the special form of a trigonometric series with an exponentially decaying function of time, based on the concept of frequency splitting. By applying harmonic balance, the parametric vibration equation is transformed into an infinite set of homogeneous linear equations, from which the principal oscillation frequency can be computed, and all coefficients of harmonic components can be obtained. With initial conditions, arbitrary constants in a general solution can be determined. To analyze the computational accuracy and consistency, an approach error function is defined, which is used to assess the computational error in the proposed approach and in the standard numerical approach based on the Runge-Kutta algorithm. Furthermore, an example of a dynamic model of airplane wing flutter on a turbine engine is given to illustrate the applicability of the proposed approach. Numerical solutions show that the proposed approach exhibits high accuracy in mathematical expression, and it is valuable for theoretical research and engineering applications of parametric systems.

Planar reorientation of a free-free beam in space using embedded electromechanical actuators

NASA Technical Reports Server (NTRS)

Kolmanovsky, Ilya V.; Mcclamroch, N. Harris

1993-01-01

It is demonstrated that the planar reorientation of a free-free beam in zero gravity space can be accomplished by periodically changing the shape of the beam using embedded electromechanical actuators. The dynamics which determine the shape of the free-free beam is assumed to be characterized by the Euler-Bernoulli equation, including material damping, with appropriate boundary conditions. The coupling between the rigid body motion and the flexible motion is explained using the angular momentum expression which includes rotatory inertia and kinematically exact effects. A control scheme is proposed where the embedded actuators excite the flexible motion of the beam so that it rotates in the desired sense with respect to a fixed inertial reference. Relations are derived which relate the average rotation rate to the amplitudes and the frequencies of the periodic actuation signal and the properties of the beam. These reorientation maneuvers can be implemented by using feedback control.

Using cadence to study free-living ambulatory behaviour.

PubMed

Tudor-Locke, Catrine; Rowe, David A

2012-05-01

The health benefits of a physically active lifestyle across a person's lifespan have been established. If there is any single physical activity behaviour that we should measure well and promote effectively, it is ambulatory activity and, more specifically, walking. Since public health physical activity guidelines include statements related to intensity of activity, it follows that we need to measure and promote free-living patterns of ambulatory activity that are congruent with this intent. The purpose of this review article is to present and summarize the potential for using cadence (steps/minute) to represent such behavioural patterns of ambulatory activity in free-living. Cadence is one of the spatio-temporal parameters of gait or walking speed. It is typically assessed using short-distance walks in clinical research and practice, but free-living cadence can be captured with a number of commercially available accelerometers that possess time-stamping technology. This presents a unique opportunity to use the same metric to communicate both ambulatory performance (assessed under testing conditions) and behaviour (assessed in the real world). Ranges for normal walking cadence assessed under laboratory conditions are 96-138 steps/minute for women and 81-135 steps/minute for men across their lifespan. The correlation between mean cadence and intensity (assessed with indirect calorimetry and expressed as metabolic equivalents [METs]) based on five treadmill/overground walking studies, is r = 0.93 and 100 steps/minute is considered to be a reasonable heuristic value indicative of walking at least at absolutely-defined moderate intensity (i.e. minimally, 3 METs) in adults. The weighted mean cadence derived from eight studies that have observed pedestrian cadence under natural conditions was 115.2 steps/minute, demonstrating that achieving 100 steps/minute is realistic in specific settings that occur in real life. However, accelerometer data collected in a large

Stress-free end problem in layered materials

NASA Technical Reports Server (NTRS)

Erdogan, F.; Bakioglu, M.

1977-01-01

In this paper the plane elastostatic problem for a medium which consists of periodically arranged two sets of bonded dissimilar layers or strips is considered. First it is assumed that one set of strips contains a crack which crosses the bimaterial interfaces. Then, by letting the collinear cracks join, the stress-free end problem is formulated. The singular behavior of the solutions at the point on intersection of the stress-free boundary and the interfaces is examined and appropriate stress intensity factors are defined. The results of some numerical examples are then presented which include the cases of both plane stress and plane strain.

Why patients visit their doctors: assessing the most prevalent conditions in a defined American population.

PubMed

St Sauver, Jennifer L; Warner, David O; Yawn, Barbara P; Jacobson, Debra J; McGree, Michaela E; Pankratz, Joshua J; Melton, L Joseph; Roger, Véronique L; Ebbert, Jon O; Rocca, Walter A

2013-01-01

To describe the prevalence of nonacute conditions among patients seeking health care in a defined US population, emphasizing age, sex, and ethnic differences. The Rochester Epidemiology Project (REP) medical records linkage system was used to identify all residents of Olmsted County, Minnesota, on April 1, 2009, who had consented to review of their medical records for research (142,377 patients). We then electronically extracted all International Classification of Diseases, Ninth Revision codes noted in the records of these patients by any health care institution between January 1, 2005, and December 31, 2009. We grouped International Classification of Diseases, Ninth Revision codes into clinical classification codes and then into 47 broader disease groups associated with health-related quality of life. Age- and sex-specific prevalence was estimated by dividing the number of individuals within each group by the corresponding age- and sex-specific population. Patients within a group who had multiple codes were counted only once. We included a total of 142,377 patients, 75,512 (53%) of whom were female. Skin disorders (42.7%), osteoarthritis and joint disorders (33.6%), back problems (23.9%), disorders of lipid metabolism (22.4%), and upper respiratory tract disease (22.1%, excluding asthma) were the most prevalent disease groups in this population. Ten of the 15 most prevalent disease groups were more common in women in almost all age groups, whereas disorders of lipid metabolism, hypertension, and diabetes were more common in men. Additionally, the prevalence of 7 of the 10 most common groups increased with advancing age. Prevalence also varied across ethnic groups (whites, blacks, and Asians). Our findings suggest areas for focused research that may lead to better health care delivery and improved population health. Copyright © 2013 Mayo Foundation for Medical Education and Research. Published by Elsevier Inc. All rights reserved.

'GREENER' CHEMICAL SYNTHESES USING MICROWAVES UNDER SOLVENT-FREE CONDITIONS OR AQUEOUS MEDIA

EPA Science Inventory

A solvent-free approach that involves microwave (MW) exposure of neat reactants (undiluted) catalyzed by the surfaces of recyclable mineral supports such as alumina, silica, clay, or 'doped' surfaces is presented which is applicable to a wide range of cleavage, condensation, cycl...

Global Free-tropospheric NO2 Abundances Derived Using a Cloud Slicing Technique from AURA OMI

NASA Technical Reports Server (NTRS)

Choi, S.; Joiner, J.; Choi, Y.; Duncan, B.N.; Vasilkov, A.; Krotkov, N.; Bucsela, E.J.

2014-01-01

We derive free-tropospheric NO2 volume mixing ratios (VMRs) by applying a cloud-slicing technique to data from the Ozone Monitoring Instrument (OMI) on the Aura satellite. In the cloud-slicing approach, the slope of the above-cloud NO2 column versus the cloud scene pressure is proportional to the NO2 VMR. In this work, we use a sample of nearby OMI pixel data from a single orbit for the linear fit. The OMI data include cloud scene pressures from the rotational-Raman algorithm and above-cloud NO2 vertical column density (VCD) (defined as the NO2 column from the cloud scene pressure to the top of the atmosphere) from a differential optical absorption spectroscopy (DOAS) algorithm. We compare OMI-derived NO2 VMRs with in situ aircraft profiles measured during the NASA Intercontinental Chemical Transport Experiment Phase B (INTEX-B) campaign in 2006. The agreement is generally within the estimated uncertainties when appropriate data screening is applied. We then derive a global seasonal climatology of free-tropospheric NO2 VMR in cloudy conditions. Enhanced NO2 in the free troposphere commonly appears near polluted urban locations where NO2 produced in the boundary layer may be transported vertically out of the boundary layer and then horizontally away from the source. Signatures of lightning NO2 are also shown throughout low and middle latitude regions in summer months. A profile analysis of our cloud-slicing data indicates signatures of lightning-generated NO2 in the upper troposphere. Comparison of the climatology with simulations from the global modeling initiative (GMI) for cloudy conditions (cloud optical depth less than10) shows similarities in the spatial patterns of continental pollution outflow. However, there are also some differences in the seasonal variation of free-tropospheric NO2 VMRs near highly populated regions and in areas affected by lightning-generated NOx.

Generating Germ-Free Drosophila to Study Gut-Microbe Interactions: Protocol to Rear Drosophila Under Axenic Conditions.

PubMed

Kietz, Christa; Pollari, Vilma; Meinander, Annika

2018-06-22

As several diseases have been linked to dysbiosis of the human intestinal microflora, manipulation of the microbiota has emerged as an exciting new strategy for potentially treating and preventing diseases. However, the human microbiota consists of a plethora of different species, and distinguishing the impact of a specific bacterial species on human health is challenging. In tackling this challenge, the fruit fly Drosophila melanogaster, with its far simpler microbial composition, has emerged as a powerful model for unraveling host-microbe interactions. To study the interplay between the resident commensal microbiome and the host, flies can be made germ-free, or axenic. To elucidate the impact of specific bacteria, axenic flies can then be re-introduced to specific microbial species. In this unit, we provide a step-by-step protocol on how to rear Drosophila melanogaster under axenic conditions and confirm the axenity of flies. © 2018 by John Wiley & Sons, Inc. © 2018 John Wiley & Sons, Inc.

19 CFR 10.102 - Duty-free entries.

Code of Federal Regulations, 2010 CFR

2010-04-01

... ARTICLES CONDITIONALLY FREE, SUBJECT TO A REDUCED RATE, ETC. General Provisions United States Government... director shall only admit articles free of duty under subheadings 9808.00.30, 9808.00.40, 9808.00.50, HTSUS... articles under subheadings 9808.00.30, 9808.00.40, or 9808.00.50, HTSUS (19 U.S.C. 1202). The certificates...

Effective matrix-free preconditioning for the augmented immersed interface method

NASA Astrophysics Data System (ADS)

Xia, Jianlin; Li, Zhilin; Ye, Xin

2015-12-01

We present effective and efficient matrix-free preconditioning techniques for the augmented immersed interface method (AIIM). AIIM has been developed recently and is shown to be very effective for interface problems and problems on irregular domains. GMRES is often used to solve for the augmented variable(s) associated with a Schur complement A in AIIM that is defined along the interface or the irregular boundary. The efficiency of AIIM relies on how quickly the system for A can be solved. For some applications, there are substantial difficulties involved, such as the slow convergence of GMRES (particularly for free boundary and moving interface problems), and the inconvenience in finding a preconditioner (due to the situation that only the products of A and vectors are available). Here, we propose matrix-free structured preconditioning techniques for AIIM via adaptive randomized sampling, using only the products of A and vectors to construct a hierarchically semiseparable matrix approximation to A. Several improvements over existing schemes are shown so as to enhance the efficiency and also avoid potential instability. The significance of the preconditioners includes: (1) they do not require the entries of A or the multiplication of AT with vectors; (2) constructing the preconditioners needs only O (log ⁡ N) matrix-vector products and O (N) storage, where N is the size of A; (3) applying the preconditioners needs only O (N) flops; (4) they are very flexible and do not require any a priori knowledge of the structure of A. The preconditioners are observed to significantly accelerate the convergence of GMRES, with heuristical justifications of the effectiveness. Comprehensive tests on several important applications are provided, such as Navier-Stokes equations on irregular domains with traction boundary conditions, interface problems in incompressible flows, mixed boundary problems, and free boundary problems. The preconditioning techniques are also useful for

A Comparison of the Scalar and Vorticity Criterion defining the T/NT Interface

NASA Astrophysics Data System (ADS)

Boschung, Jonas; Hennig, Fabian; Peters, Norbert

2013-11-01

Free shear flows are characterized by a turbulent core region, a non-turbulent outer flow and a turbulent/non-turbulent interface separating the two zones. While there exist different approaches to identify this transitional region, the interface position is mostly defined to coincide with the isoscalar surfaces of either a passive scalar or the magnitude of the vorticity. Both criteria are examined and compared using a shear layer DNS.

Piper-betle-shaped nano-S-catalyzed synthesis of 1-amidoalkyl-2-naphthols under solvent-free reaction condition: a greener "nanoparticle-catalyzed organic synthesis enhancement" approach.

PubMed

Das, Vijay K; Borah, Madhurjya; Thakur, Ashim J

2013-04-05

Nano-S prepared by an annealing process showed excellent catalytic activity for the synthesis of 1-amidoalkyl-2-naphthols under solvent-free reaction condition at 50 °C. The catalyst could be reused up to the fifth cycle without loss in its action. The green-ness of the present protocol was also measured using green metrics drawing its superiority.

Wingtip Vortices and Free Shear Layer Interaction in the Vicinity of Maximum Lift to Drag Ratio Lift Condition

NASA Astrophysics Data System (ADS)

Memon, Muhammad Omar

Cost-effective air-travel is something everyone wishes for when it comes to booking flights. The continued and projected increase in commercial air travel advocates for energy efficient airplanes, reduced carbon footprint, and a strong need to accommodate more airplanes into airports. All of these needs are directly affected by the magnitudes of drag these aircraft experience and the nature of their wingtip vortex. A large portion of the aerodynamic drag results from the airflow rolling from the higher pressure side of the wing to the lower pressure side, causing the wingtip vortices. The generation of this particular drag is inevitable however, a more fundamental understanding of the phenomenon could result in applications whose benefits extend much beyond the relatively minuscule benefits of commonly-used winglets. Maximizing airport efficiency calls for shorter intervals between takeoffs and landings. Wingtip vortices can be hazardous for following aircraft that may fly directly through the high-velocity swirls causing upsets at vulnerably low speeds and altitudes. The vortex system in the near wake is typically more complex since strong vortices tend to continue developing throughout the near wake region. Several chord lengths distance downstream of a wing, the so-called fully rolled up wing wake evolves into a combination of a discrete wingtip vortex pair and a free shear layer. Lift induced drag is generated as a byproduct of downwash induced by the wingtip vortices. The parasite drag results from a combination of form/pressure drag and the upper and lower surface boundary layers. These parasite effects amalgamate to create the free shear layer in the wake. While the wingtip vortices embody a large portion of the total drag at lifting angles, flow properties in the free shear layer also reveal their contribution to the aerodynamic efficiency of the aircraft. Since aircraft rarely cruise at maximum aerodynamic efficiency, a better understanding of the balance

The validity of consumer-level, activity monitors in healthy adults worn in free-living conditions: a cross-sectional study.

PubMed

Ferguson, Ty; Rowlands, Alex V; Olds, Tim; Maher, Carol

2015-03-27

Technological advances have seen a burgeoning industry for accelerometer-based wearable activity monitors targeted at the consumer market. The purpose of this study was to determine the convergent validity of a selection of consumer-level accelerometer-based activity monitors. 21 healthy adults wore seven consumer-level activity monitors (Fitbit One, Fitbit Zip, Jawbone UP, Misfit Shine, Nike Fuelband, Striiv Smart Pedometer and Withings Pulse) and two research-grade accelerometers/multi-sensor devices (BodyMedia SenseWear, and ActiGraph GT3X+) for 48-hours. Participants went about their daily life in free-living conditions during data collection. The validity of the consumer-level activity monitors relative to the research devices for step count, moderate to vigorous physical activity (MVPA), sleep and total daily energy expenditure (TDEE) was quantified using Bland-Altman analysis, median absolute difference and Pearson's correlation. All consumer-level activity monitors correlated strongly (r > 0.8) with research-grade devices for step count and sleep time, but only moderately-to-strongly for TDEE (r = 0.74-0.81) and MVPA (r = 0.52-0.91). Median absolute differences were generally modest for sleep and steps (<10% of research device mean values for the majority of devices) moderate for TDEE (<30% of research device mean values), and large for MVPA (26-298%). Across the constructs examined, the Fitbit One, Fitbit Zip and Withings Pulse performed most strongly. In free-living conditions, the consumer-level activity monitors showed strong validity for the measurement of steps and sleep duration, and moderate valid for measurement of TDEE and MVPA. Validity for each construct ranged widely between devices, with the Fitbit One, Fitbit Zip and Withings Pulse being the strongest performers.

Sensorimotor-Conceptual Integration in Free Walking Enhances Divergent Thinking for Young and Older Adults

PubMed Central

Kuo, Chun-Yu; Yeh, Yei-Yu

2016-01-01

Prior research has shown that free walking can enhance creative thinking. Nevertheless, it remains unclear whether bidirectional body-mind links are essential for the positive effect of free walking on creative thinking. Moreover, it is unknown whether the positive effect can be generalized to older adults. In Experiment 1, we replicated previous findings with two additional groups of young participants. Participants in the rectangular-walking condition walked along a rectangular path while generating unusual uses for chopsticks. Participants in the free-walking group walked freely as they wished, and participants in the free-generation condition generated unconstrained free paths while the participants in the random-experienced condition walked those paths. Only the free-walking group showed better performance in fluency, flexibility, and originality. In Experiment 2, two groups of older adults were randomly assigned to the free-walking and rectangular-walking conditions. The free-walking group showed better performance than the rectangular-walking group. Moreover, older adults in the free-walking group outperformed young adults in the rectangular-walking group in originality and performed comparably in fluency and flexibility. Bidirectional links between proprioceptive-motor kinematics and metaphorical abstract concepts can enhance divergent thinking for both young and older adults. PMID:27790178

How main-chains of proteins explore the free-energy landscape in native states.

PubMed

Senet, Patrick; Maisuradze, Gia G; Foulie, Colette; Delarue, Patrice; Scheraga, Harold A

2008-12-16

Understanding how a single native protein diffuses on its free-energy landscape is essential to understand protein kinetics and function. The dynamics of a protein is complex, with multiple relaxation times reflecting a hierarchical free-energy landscape. Using all-atom molecular dynamics simulations of an alpha/beta protein (crambin) and a beta-sheet polypeptide (BS2) in their "native" states, we demonstrate that the mean-square displacement of dihedral angles, defined by 4 successive C(alpha) atoms, increases as a power law of time, t(alpha), with an exponent alpha between 0.08 and 0.39 (alpha = 1 corresponds to Brownian diffusion), at 300 K. Residues with low exponents are located mainly in well-defined secondary elements and adopt 1 conformational substate. Residues with high exponents are found in loops/turns and chain ends and exist in multiple conformational substates, i.e., they move on multiple-minima free-energy profiles.

Functionalized SBA-15 supported nickel (II)-oxime-imine catalysts for liquid phase oxidation of olefins under solvent-free conditions

NASA Astrophysics Data System (ADS)

Paul, Luna; Banerjee, Biplab; Bhaumik, Asim; Ali, Mahammad

2016-05-01

A new oxime-imine functionalized highly ordered mesoporous SBA-15 (SBA-15-NH2-DAMO) has been synthesized via post-synthesis functionalization of SBA-15 with 3-aminopropyl-triethoxysilane followed by the Schiff base condensation with diacetylmonooxime, which was further reacted with Ni(ClO4)2 to yield the functionalized nickel catalyst SBA-15-NH2-DAMO-Ni. All the synthesized materials were thoroughly characterized using different characterization techniques. It was found that SBA-15-NH2-DAMO-Ni catalyzes the one-pot oxidation of olefins like styrene, cyclohexene, cyclooctene, 1-hexene and 1-octene to the corresponding benzaldehyde, cyclohexene-1-ol and cyclooctene-oxide, respectively under solvent-free conditions by using tert-butylhydroperoxide as oxidant.

Defining human death: an intersection of bioethics and metaphysics.

PubMed

Manninen, Bertha Alvarez

2009-01-01

For many years now, bioethicists, physicians, and others in the medical field have disagreed concerning how to best define human death. Different theories range from the Harvard Criteria of Brain Death, which defines death as the cessation of all brain activity, to the Cognitive Criteria, which is based on the loss of almost all core mental properties, e.g., memory, self-consciousness, moral agency, and the capacity for reason. A middle ground is the Irreversibility Standard, which defines death as occurring when the capacity for consciousness is forever lost. Given all these different theories, how can we begin to approach solving the issue of how to define death? I propose that a necessary starting point is discussing an even more fundamental question that properly belongs in the philosophical field of metaphysics: we must first address the issue of diachronic identity over time, and the persistence conditions of personal identity. In this paper, I illustrate the interdependent relationship between this metaphysical question and questions concerning the definition of death. I also illustrate how it is necessary to antecedently attend to the metaphysical issue of defining death before addressing certain issues in medical ethics, e.g., whether it is morally permissible to euthanize patients in persistent vegetative states or procure organs from anencephalic infants.

Semantics of Context-Free Fragments of Natural Languages.

ERIC Educational Resources Information Center

Suppes, Patrick

The objective of this paper is to combine the viewpoint of model-theoretic semantics and generative grammar, to define semantics for context-free languages, and to apply the results to some fragments of natural language. Following the introduction in the first section, Section 2 describes a simple artificial example to illustrate how a semantic…

The four free-operant freedoms

PubMed Central

Lindsley, Ogden R.

1996-01-01

This article reviews early free-operant conditioning laboratory research and applications. The seldom-mentioned four free-operant freedoms are described for the first time in detail. Most current behavior analysts do not realize that the freedom to form responses and the freedom to speed responses were crucial steps in designing free-operant operanda in the 1950s. These four freedoms were known by the laboratory researchers of the 1950s to the point that, along with operanda design, Sidman (1960) did not feel the need to detail them in his classic, Tactics of Scientific Research. The dimensions of freedom in the operant were so well understood and accepted in the 1950s that most thought it redundant to use the term free operant. These issues are reviewed in some detail for younger behavior analysts who did not have the opportunity of learning them firsthand. PMID:22478258

Stability of Hydrocortisone Preservative-Free Oral Solutions.

PubMed

Chappe, Julie; Osman, Névine; Cisternino, Salvatore; Fontan, Jean-Eudes; Schlatter, Joël

2015-01-01

The physical and chemical stability of a preservative-free oral solution of hydrocortisone succinate was studied at different pH values and storage temperatures. Oral solutions of hydrocortisone 1 mg/mL were prepared by dissolving hydrocortisone succinate powder in citrate buffers at pH 4.0, 5.5, and 6.5, or with sterile water (pH 7.4) stored in amber glass vials. Three identical samples of the formulations were prepared and stored under refrigeration (3-7°C), ambient temperature (20-22°C) and high temperature (29-31°C). A 200-μL sample was withdrawn from each of the 3 samples immediately after preparation and at 1, 7, 14, 21, and 35 days. Samples were assayed in duplicate using stability-indicating liquid chromatography. Stability was determined by evaluating the percentage of the initial concentration remaining at each time point; stability was defined as the retention of at least 90% of the initial concentration of hydrocortisone succinate. At least 92% of the initial hydrocortisone succinate concentration in solutions pH 5.5, 6.5, and 7.4 remained throughout the 14-day study period under refrigeration. There were no detectable changes in color, odor, or pH and no visible microbial growth in these samples. In other storage conditions, hydrocortisone succinate was rapidly degraded. The hydrocortisone succinate preservative-free oral solutions at pH 5.5, 6.5, or 7.4 are chemically stable when stored under refrigeration for at least 14 days. They provide flexible and convenient dosage forms without any preservatives for pediatric patients.

Context-free parsing with connectionist networks

NASA Astrophysics Data System (ADS)

Fanty, M. A.

1986-08-01

This paper presents a simple algorithm which converts any context-free grammar into a connectionist network which parses strings (of arbitrary but fixed maximum length) in the language defined by that grammar. The network is fast, O(n), and deterministicd. It consists of binary units which compute a simple function of their input. When the grammar is put in Chomsky normal form, O(n3) units needed to parse inputs of length up to n.

Non-Euclidean stress-free configuration of arteries accounting for curl of axial strips sectioned from vessels.

PubMed

Takamizawa, Keiichi; Nakayama, Yasuhide

2013-11-01

It is well known that arteries are subject to residual stress. In earlier studies, the residual stress in the arterial ring relieved by a radial cut was considered in stress analysis. However, it has been found that axial strips sectioned from arteries also curled into arcs, showing that the axial residual stresses were relieved from the arterial walls. The combined relief of circumferential and axial residual stresses must be considered to accurately analyze stress and strain distributions under physiological loading conditions. In the present study, a mathematical model of a stress-free configuration of artery was proposed using Riemannian geometry. Stress analysis for arterial walls under unloaded and physiologically loaded conditions was performed using exponential strain energy functions for porcine and human common carotid arteries. In the porcine artery, the circumferential stress distribution under physiological loading became uniform compared with that without axial residual strain, whereas a gradient of axial stress distribution increased through the wall thickness. This behavior showed almost the same pattern that was observed in a recent study in which approximate analysis accounting for circumferential and axial residual strains was performed, whereas the circumferential and axial stresses increased from the inner surface to the outer surface under a physiological condition in the human common carotid artery of a two-layer model based on data of other recent studies. In both analyses, Riemannian geometry was appropriate to define the stress-free configurations of the arterial walls with both circumferential and axial residual strains.

Equivalence principle for quantum systems: dephasing and phase shift of free-falling particles

NASA Astrophysics Data System (ADS)

Anastopoulos, C.; Hu, B. L.

2018-02-01

We ask the question of how the (weak) equivalence principle established in classical gravitational physics should be reformulated and interpreted for massive quantum objects that may also have internal degrees of freedom (dof). This inquiry is necessary because even elementary concepts like a classical trajectory are not well defined in quantum physics—trajectories originating from quantum histories become viable entities only under stringent decoherence conditions. From this investigation we posit two logically and operationally distinct statements of the equivalence principle for quantum systems. Version A: the probability distribution of position for a free-falling particle is the same as the probability distribution of a free particle, modulo a mass-independent shift of its mean. Version B: any two particles with the same velocity wave-function behave identically in free fall, irrespective of their masses. Both statements apply to all quantum states, including those without a classical correspondence, and also for composite particles with quantum internal dof. We also investigate the consequences of the interaction between internal and external dof induced by free fall. For a class of initial states, we find dephasing occurs for the translational dof, namely, the suppression of the off-diagonal terms of the density matrix, in the position basis. We also find a gravitational phase shift in the reduced density matrix of the internal dof that does not depend on the particle’s mass. For classical states, the phase shift has a natural classical interpretation in terms of gravitational red-shift and special relativistic time-dilation.

Self-defining memories during exposure to music in Alzheimer's disease.

PubMed

El Haj, Mohamad; Antoine, Pascal; Nandrino, Jean Louis; Gély-Nargeot, Marie-Christine; Raffard, Stéphane

2015-10-01

Research suggests that exposure to music may enhance autobiographical recall in Alzheimer's Disease (AD) patients. This study investigated whether exposure to music could enhance the production of self-defining memories, that is, memories that contribute to self-discovery, self-understanding, and identity in AD patients. Twenty-two mild-stage AD patients and 24 healthy controls were asked to produce autobiographical memories in silence, while listening to researcher-chosen music, and to their own-chosen music. AD patients showed better autobiographical recall when listening to their own-chosen music than to researcher-chosen music or than in silence. More precisely, they produced more self-defining memories during exposure to their own-chosen music than to researcher-chosen music or during silence. Additionally, AD patients produced more self-defining memories than autobiographical episodes or personal-semantics during exposure to their own-chosen music. This pattern contrasted with the poor production of self-defining memories during silence or during exposure to researcher-chosen music. Healthy controls did not seem to enjoy the same autobiographical benefits nor the same self-defining memory enhancement in the self-chosen music condition. Poor production of self-defining memories, as observed in AD, may somehow be alleviated by exposure to self-chosen music.

Free markets and the marriage market: structural adjustment, gender relations, and working conditions among Dominican women workers.

PubMed

Safa, H I

1999-02-01

The relationship between economic conditions and marriage patterns in the Dominican Republic is analyzed using data collected in 1994 on women working in a free-trade zone. The author concludes that changes associated with structural adjustment appear to have contributed to a deterioration of the job market and a greater prevalence of female-headed households. It is shown that "structural adjustment increases the need for women to work, because of cuts in government programs, declining real wages, growing inflation, and a deterioration in male employment, which weakens the man's role as principal breadwinner and increases the importance and visibility of women's contribution to the household economy. This change in the gender composition of the labor force has encouraged some women to resist marriage and/or remarriage because the ¿marriage market' of eligible men willing and able to support a family has been reduced, contributing to greater marital instability." excerpt

PIES free boundary stellarator equilibria with improved initial conditions

NASA Astrophysics Data System (ADS)

Drevlak, M.; Monticello, D.; Reiman, A.

2005-07-01

The MFBE procedure developed by Strumberger (1997 Nucl. Fusion 37 19) is used to provide an improved starting point for free boundary equilibrium computations in the case of W7-X (Nührenberg and Zille 1986 Phys. Lett. A 114 129) using the Princeton iterative equilibrium solver (PIES) code (Reiman and Greenside 1986 Comput. Phys. Commun. 43 157). Transferring the consistent field found by the variational moments equilibrium code (VMEC) (Hirshmann and Whitson 1983 Phys. Fluids 26 3553) to an extended coordinate system using the VMORPH code, a safe margin between plasma boundary and PIES domain is established. The new EXTENDER_P code implements a generalization of the virtual casing principle, which allows field extension both for VMEC and PIES equilibria. This facilitates analysis of the 5/5 islands of the W7-X standard case without including them in the original PIES computation.

The effects of electrospun substrate-mediated cell colony morphology on the self-renewal of human induced pluripotent stem cells.

PubMed

Maldonado, Maricela; Wong, Lauren Y; Echeverria, Cristina; Ico, Gerardo; Low, Karen; Fujimoto, Taylor; Johnson, Jed K; Nam, Jin

2015-05-01

The development of xeno-free, chemically defined stem cell culture systems has been a primary focus in the field of regenerative medicine to enhance the clinical application of pluripotent stem cells (PSCs). In this regard, various electrospun substrates with diverse physiochemical properties were synthesized utilizing various polymer precursors and surface treatments. Human induced pluripotent stem cells (IPSCs) cultured on these substrates were characterized by their gene and protein expression to determine the effects of the substrate physiochemical properties on the cells' self-renewal, i.e., proliferation and the maintenance of pluripotency. The results showed that surface chemistry significantly affected cell colony formation via governing the colony edge propagation. More importantly, when surface chemistry of the substrates was uniformly controlled by collagen conjugation, the stiffness of substrate was inversely related to the sphericity, a degree of three dimensionality in colony morphology. The differences in sphericity subsequently affected spontaneous differentiation of IPSCs during a long-term culture, implicating that the colony morphology is a deciding factor in the lineage commitment of PSCs. Overall, we show that the capability of controlling IPSC colony morphology by electrospun substrates provides a means to modulate IPSC self-renewal. Copyright © 2015 Elsevier Ltd. All rights reserved.

Arraying proteins by cell-free synthesis.

PubMed

He, Mingyue; Wang, Ming-Wei

2007-10-01

Recent advances in life science have led to great motivation for the development of protein arrays to study functions of genome-encoded proteins. While traditional cell-based methods have been commonly used for generating protein arrays, they are usually a time-consuming process with a number of technical challenges. Cell-free protein synthesis offers an attractive system for making protein arrays, not only does it rapidly converts the genetic information into functional proteins without the need for DNA cloning, but also presents a flexible environment amenable to production of folded proteins or proteins with defined modifications. Recent advancements have made it possible to rapidly generate protein arrays from PCR DNA templates through parallel on-chip protein synthesis. This article reviews current cell-free protein array technologies and their proteomic applications.

Free and Forced Vibrations of Thick-Walled Anisotropic Cylindrical Shells

NASA Astrophysics Data System (ADS)

Marchuk, A. V.; Gnedash, S. V.; Levkovskii, S. A.

2017-03-01

Two approaches to studying the free and forced axisymmetric vibrations of cylindrical shell are proposed. They are based on the three-dimensional theory of elasticity and division of the original cylindrical shell with concentric cross-sectional circles into several coaxial cylindrical shells. One approach uses linear polynomials to approximate functions defined in plan and across the thickness. The other approach also uses linear polynomials to approximate functions defined in plan, but their variation with thickness is described by the analytical solution of a system of differential equations. Both approaches have approximation and arithmetic errors. When determining the natural frequencies by the semi-analytical finite-element method in combination with the divide and conqure method, it is convenient to find the initial frequencies by the finite-element method. The behavior of the shell during free and forced vibrations is analyzed in the case where the loading area is half the shell thickness

How main-chains of proteins explore the free-energy landscape in native states

PubMed Central

Senet, Patrick; Maisuradze, Gia G.; Foulie, Colette; Delarue, Patrice; Scheraga, Harold A.

2008-01-01

Understanding how a single native protein diffuses on its free-energy landscape is essential to understand protein kinetics and function. The dynamics of a protein is complex, with multiple relaxation times reflecting a hierarchical free-energy landscape. Using all-atom molecular dynamics simulations of an α/β protein (crambin) and a β-sheet polypeptide (BS2) in their “native” states, we demonstrate that the mean-square displacement of dihedral angles, defined by 4 successive Cα atoms, increases as a power law of time, tα, with an exponent α between 0.08 and 0.39 (α = 1 corresponds to Brownian diffusion), at 300 K. Residues with low exponents are located mainly in well-defined secondary elements and adopt 1 conformational substate. Residues with high exponents are found in loops/turns and chain ends and exist in multiple conformational substates, i.e., they move on multiple-minima free-energy profiles. PMID:19073932

Cluster Free Energies from Simple Simulations of Small Numbers of Aggregants: Nucleation of Liquid MTBE from Vapor and Aqueous Phases.

PubMed

Patel, Lara A; Kindt, James T

2017-03-14

We introduce a global fitting analysis method to obtain free energies of association of noncovalent molecular clusters using equilibrated cluster size distributions from unbiased constant-temperature molecular dynamics (MD) simulations. Because the systems simulated are small enough that the law of mass action does not describe the aggregation statistics, the method relies on iteratively determining a set of cluster free energies that, using appropriately weighted sums over all possible partitions of N monomers into clusters, produces the best-fit size distribution. The quality of these fits can be used as an objective measure of self-consistency to optimize the cutoff distance that determines how clusters are defined. To showcase the method, we have simulated a united-atom model of methyl tert-butyl ether (MTBE) in the vapor phase and in explicit water solution over a range of system sizes (up to 95 MTBE in the vapor phase and 60 MTBE in the aqueous phase) and concentrations at 273 K. The resulting size-dependent cluster free energy functions follow a form derived from classical nucleation theory (CNT) quite well over the full range of cluster sizes, although deviations are more pronounced for small cluster sizes. The CNT fit to cluster free energies yielded surface tensions that were in both cases lower than those for the simulated planar interfaces. We use a simple model to derive a condition for minimizing non-ideal effects on cluster size distributions and show that the cutoff distance that yields the best global fit is consistent with this condition.

[Iron and invasive fungal infection].

PubMed

Álvarez, Florencio; Fernández-Ruiz, Mario; Aguado, José María

2013-01-01

Iron is an essential factor for both the growth and virulence of most of microorganisms. As a part of the innate (or nutritional) immune system, mammals have developed different mechanisms to store and transport this element in order to limit free iron bioavailability. To survive in this hostile environment, pathogenic fungi have specific uptake systems for host iron sources, one of the most important of which is based on the synthesis of siderophores-soluble, low-molecular-mass, high-affinity iron chelators. The increase in free iron that results from iron-overload conditions is a well-established risk factor for invasive fungal infection (IFI) such as mucormycosis or aspergillosis. Therefore, iron chelation may be an appealing therapeutic option for these infections. Nevertheless, deferoxamine -the first approved iron chelator- paradoxically increases the incidence of IFI, as it serves as a xeno-siderophore to Mucorales. On the contrary, the new oral iron chelators (deferiprone and deferasirox) have shown to exert a deleterious effect on fungal growth both in vitro and in animal models. The present review focuses on the role of iron metabolism in the pathogenesis of IFI and summarises the preclinical data, as well as the limited clinical experience so far, in the use of new iron chelators as treatment for mucormycosis and invasive aspergillosis. Copyright © 2012 Revista Iberoamericana de Micología. Published by Elsevier Espana. All rights reserved.

Clinical-scale validation of a new efficient procedure for cryopreservation of ex vivo expanded cord blood hematopoietic stem and progenitor cells.

PubMed

Duchez, Pascale; Rodriguez, Laura; Chevaleyre, Jean; De La Grange, Philippe Brunet; Ivanovic, Zoran

2016-12-01

Survival of ex vivo expanded hematopoietic stem cells (HSC) and progenitor cells is low with the standard cryopreservation procedure. We recently showed that the efficiency of cryopreservation of these cells may be greatly enhanced by adding a serum-free xeno-free culture medium (HP01 Macopharma), which improves the antioxidant and biochemical properties of the cryopreservation solution. Here we present the clinical-scale validation of this cryopreservation procedure. The hematopoietic cells expanded in clinical-scale cultures were cryopreserved applying the new HP01-based procedure. The viability, apoptosis rate and number of functional committed progenitors (methyl-cellulose colony forming cell test), short-term repopulating HSCs (primary recipient NSG mice) and long-term HSCs (secondary recipient NSG mice) were tested before and after thawing. The efficiency of clinical-scale procedure reproduced the efficiency of cryopreservation obtained earlier in miniature sample experiments. Furthermore, the full preservation of short- and long-term HSCs was obtained in clinical scale conditions. Because the results obtained in clinical-scale volume are comparable to our earlier results in miniature-scale cultures, the clinical-scale procedure should be considered validated. It allows cryopreservation of the whole ex vivo expanded culture content, conserving full short- and long-term HSC activity. Copyright © 2016 International Society for Cellular Therapy. Published by Elsevier Inc. All rights reserved.

MHD Free Convective Boundary Layer Flow of a Nanofluid past a Flat Vertical Plate with Newtonian Heating Boundary Condition

PubMed Central

Uddin, Mohammed J.; Khan, Waqar A.; Ismail, Ahmed I.

2012-01-01

Steady two dimensional MHD laminar free convective boundary layer flows of an electrically conducting Newtonian nanofluid over a solid stationary vertical plate in a quiescent fluid taking into account the Newtonian heating boundary condition is investigated numerically. A magnetic field can be used to control the motion of an electrically conducting fluid in micro/nano scale systems used for transportation of fluid. The transport equations along with the boundary conditions are first converted into dimensionless form and then using linear group of transformations, the similarity governing equations are developed. The transformed equations are solved numerically using the Runge-Kutta-Fehlberg fourth-fifth order method with shooting technique. The effects of different controlling parameters, namely, Lewis number, Prandtl number, buoyancy ratio, thermophoresis, Brownian motion, magnetic field and Newtonian heating on the flow and heat transfer are investigated. The numerical results for the dimensionless axial velocity, temperature and nanoparticle volume fraction as well as the reduced Nusselt and Sherwood number have been presented graphically and discussed. It is found that the rate of heat and mass transfer increase as Newtonian heating parameter increases. The dimensionless velocity and temperature distributions increase with the increase of Newtonian heating parameter. The results of the reduced heat transfer rate is compared for convective heating boundary condition and found an excellent agreement. PMID:23166688

Free will beliefs predict attitudes toward unethical behavior and criminal punishment

PubMed Central

Martin, Nathan D.; Rigoni, Davide; Vohs, Kathleen D.

2017-01-01

Do free will beliefs influence moral judgments? Answers to this question from theoretical and empirical perspectives are controversial. This study attempted to replicate past research and offer theoretical insights by analyzing World Values Survey data from residents of 46 countries (n = 65,111 persons). Corroborating experimental findings, free will beliefs predicted intolerance of unethical behaviors and support for severe criminal punishment. Further, the link between free will beliefs and intolerance of unethical behavior was moderated by variations in countries’ institutional integrity, defined as the degree to which countries had accountable, corruption-free public sectors. Free will beliefs predicted intolerance of unethical behaviors for residents of countries with high and moderate institutional integrity, but this correlation was not seen for countries with low institutional integrity. Free will beliefs predicted support for criminal punishment regardless of countries’ institutional integrity. Results were robust across different operationalizations of institutional integrity and with or without statistical control variables. PMID:28652361

Free will beliefs predict attitudes toward unethical behavior and criminal punishment.

PubMed

Martin, Nathan D; Rigoni, Davide; Vohs, Kathleen D

2017-07-11

Do free will beliefs influence moral judgments? Answers to this question from theoretical and empirical perspectives are controversial. This study attempted to replicate past research and offer theoretical insights by analyzing World Values Survey data from residents of 46 countries ( n = 65,111 persons). Corroborating experimental findings, free will beliefs predicted intolerance of unethical behaviors and support for severe criminal punishment. Further, the link between free will beliefs and intolerance of unethical behavior was moderated by variations in countries' institutional integrity, defined as the degree to which countries had accountable, corruption-free public sectors. Free will beliefs predicted intolerance of unethical behaviors for residents of countries with high and moderate institutional integrity, but this correlation was not seen for countries with low institutional integrity. Free will beliefs predicted support for criminal punishment regardless of countries' institutional integrity. Results were robust across different operationalizations of institutional integrity and with or without statistical control variables.

Computing the Free Energy along a Reaction Coordinate Using Rigid Body Dynamics.

PubMed

Tao, Peng; Sodt, Alexander J; Shao, Yihan; König, Gerhard; Brooks, Bernard R

2014-10-14

The calculations of potential of mean force along complex chemical reactions or rare events pathways are of great interest because of their importance for many areas in chemistry, molecular biology, and material science. The major difficulty for free energy calculations comes from the great computational cost for adequate sampling of the system in high-energy regions, especially close to the reaction transition state. Here, we present a method, called FEG-RBD, in which the free energy gradients were obtained from rigid body dynamics simulations. Then the free energy gradients were integrated along a reference reaction pathway to calculate free energy profiles. In a given system, the reaction coordinates defining a subset of atoms (e.g., a solute, or the quantum mechanics (QM) region of a quantum mechanics/molecular mechanics simulation) are selected to form a rigid body during the simulation. The first-order derivatives (gradients) of the free energy with respect to the reaction coordinates are obtained through the integration of constraint forces within the rigid body. Each structure along the reference reaction path is separately subjected to such a rigid body simulation. The individual free energy gradients are integrated along the reference pathway to obtain the free energy profile. Test cases provided demonstrate both the strengths and weaknesses of the FEG-RBD method. The most significant benefit of this method comes from the fast convergence rate of the free energy gradient using rigid-body constraints instead of restraints. A correction to the free energy due to approximate relaxation of the rigid-body constraint is estimated and discussed. A comparison with umbrella sampling using a simple test case revealed the improved sampling efficiency of FEG-RBD by a factor of 4 on average. The enhanced efficiency makes this method effective for calculating the free energy of complex chemical reactions when the reaction coordinate can be unambiguously defined by a

Computing the Free Energy along a Reaction Coordinate Using Rigid Body Dynamics

PubMed Central

2015-01-01

The calculations of potential of mean force along complex chemical reactions or rare events pathways are of great interest because of their importance for many areas in chemistry, molecular biology, and material science. The major difficulty for free energy calculations comes from the great computational cost for adequate sampling of the system in high-energy regions, especially close to the reaction transition state. Here, we present a method, called FEG-RBD, in which the free energy gradients were obtained from rigid body dynamics simulations. Then the free energy gradients were integrated along a reference reaction pathway to calculate free energy profiles. In a given system, the reaction coordinates defining a subset of atoms (e.g., a solute, or the quantum mechanics (QM) region of a quantum mechanics/molecular mechanics simulation) are selected to form a rigid body during the simulation. The first-order derivatives (gradients) of the free energy with respect to the reaction coordinates are obtained through the integration of constraint forces within the rigid body. Each structure along the reference reaction path is separately subjected to such a rigid body simulation. The individual free energy gradients are integrated along the reference pathway to obtain the free energy profile. Test cases provided demonstrate both the strengths and weaknesses of the FEG-RBD method. The most significant benefit of this method comes from the fast convergence rate of the free energy gradient using rigid-body constraints instead of restraints. A correction to the free energy due to approximate relaxation of the rigid-body constraint is estimated and discussed. A comparison with umbrella sampling using a simple test case revealed the improved sampling efficiency of FEG-RBD by a factor of 4 on average. The enhanced efficiency makes this method effective for calculating the free energy of complex chemical reactions when the reaction coordinate can be unambiguously defined by a

Predictable dental rehabilitation in maxillomandibular reconstruction with free flaps. The role of implant guided surgery

PubMed Central

Cebrian-Carretero, José L.; Sobrino, José A.; Yu, Tomás; Burgueño-García, Miguel

2014-01-01

The reconstruction of maxillomandibular defects secondary to oral cancer surgery, represent a great challenge for Maxillofacial surgeons. During the last decades the reconstructive surgery has experimented a big advance due to the development of the microsurgical techniques. At present, we are able to reconstruct complex defects using free flaps that provide both soft and bone tissue. Fibula, iliac crest and scapula free flaps have been the three classic options for the maxillomandibular reconstruction owing to the amount of bone that this flaps provide, allowing the posterior dental rehabilitation with implants. Today, our objective it is not only the aesthetic reconstruction, but also the functional reconstruction of the patients enhancing their life quality. Guided implant surgery in free flap reconstructed patients has become an essential tool, helping to define the exact position of the dental implant in the flap. In this way it is possible to look for the areas with better bone conditions, avoiding the osteosynthesis material used to fixate the flap with the native bone and deciding the best biomechanical option, in terms of number and situation of the implants, for the future dental prostheses. In summary, using the guided implant surgery, it is possible to design an exact and predictable dental implant rehabilitation in patients with oral cancer who are reconstructed with free microvascular flap, resulting in an optimal aesthetic and functional result. Key words:Oral cancer, mandibulectomy, maxillectomy, microvascular reconstruction, fibula flap, dental implant, guided surgery. PMID:25129241

Functional characterization of human pluripotent stem cell-derived arterial endothelial cells.

PubMed

Zhang, Jue; Chu, Li-Fang; Hou, Zhonggang; Schwartz, Michael P; Hacker, Timothy; Vickerman, Vernella; Swanson, Scott; Leng, Ning; Nguyen, Bao Kim; Elwell, Angela; Bolin, Jennifer; Brown, Matthew E; Stewart, Ron; Burlingham, William J; Murphy, William L; Thomson, James A

2017-07-25

Here, we report the derivation of arterial endothelial cells from human pluripotent stem cells that exhibit arterial-specific functions in vitro and in vivo. We combine single-cell RNA sequencing of embryonic mouse endothelial cells with an EFNB2-tdTomato/EPHB4-EGFP dual reporter human embryonic stem cell line to identify factors that regulate arterial endothelial cell specification. The resulting xeno-free protocol produces cells with gene expression profiles, oxygen consumption rates, nitric oxide production levels, shear stress responses, and TNFα-induced leukocyte adhesion rates characteristic of arterial endothelial cells. Arterial endothelial cells were robustly generated from multiple human embryonic and induced pluripotent stem cell lines and have potential applications for both disease modeling and regenerative medicine.

Classic conditioning in aged rabbits: delay, trace, and long-delay conditioning.

PubMed

Solomon, P R; Groccia-Ellison, M E

1996-06-01

Young (0.5 years) and aged (2+, 3+, and 4+ years) rabbits underwent acquisition of the classically conditioned nictitating membrane response in a delay (500-ms conditioned stimulus [CS], 400-ms interstimulus interval [ISI]), long-delay (1,000-ms CS, 900-ms ISI), or trace (500-ms CS, 400-ms stimulus-free period) paradigm. Collapsing across age groups, there is a general tendency for animals to acquire trace conditioning more slowly than delay conditioning. Collapsing across conditioning paradigms, there is a general tendency for aged animals to acquire more slowly than younger animals. Of greater significance, however, are the age differences in the different conditioning paradigms. In the delay and long-delay paradigms, significant conditioning deficits first appeared in the 4(+)-year-old group. In the trace conditioning paradigm, significant conditioning deficits became apparent in the 2(+)-year-old animals.

Drinking Levels Defined

MedlinePlus

... Is A Standard Drink? Drinking Levels Defined Drinking Levels Defined Moderate alcohol consumption: According to the "Dietary ... of drinking that brings blood alcohol concentration (BAC) levels to 0.08 g/dL. This typically occurs ...

New free radicals to measure antiradical capacity: a theoretical study.

PubMed

León-Carmona, Jorge Rafael; Martínez, Ana; Galano, Annia

2014-08-28

A new family of free radicals, that are soluble in water and stable at all pH values, were recently synthesized and used to assess the antiradical capacity of several polyphenols. In the present work, density functional calculations were used to investigate the single electron transfer reactions between these new free radicals and polyphenols in aqueous solution. The quantification of the antiradical capacity is a challenge, particularly for polyphenols, since they become unstable under experimental conditions. It was found that the electron transfer from polyphenols to the newly developed free radicals can be used to assess the efficiency of this kind of compound for preventing oxidative stress. Since one of the free radicals can be deprotonated under experimental conditions, this newly synthesized radical can help distinguish more clearly between different antiradical compounds with similar antioxidant capacity by modifying the pH in the experiments. The results reported here are in good agreement with the available experimental data and allowed making recommendations about possible experimental conditions in the design of antioxidant assays using the investigated radicals.

Free vibration of rectangular plates with a small initial curvature

NASA Technical Reports Server (NTRS)

Adeniji-Fashola, A. A.; Oyediran, A. A.

1988-01-01

The method of matched asymptotic expansions is used to solve the transverse free vibration of a slightly curved, thin rectangular plate. Analytical results for natural frequencies and mode shapes are presented in the limit when the dimensionless bending rigidity, epsilon, is small compared with in-plane forces. Results for different boundary conditions are obtained when the initial deflection is: (1) a polynomial in both directions, and (2) the product of a polynomial and a trigonometric function, and arbitrary. For the arbitrary initial deflection case, the Fourier series technique is used to define the initial deflection. The results obtained show that the natural frequencies of vibration of slightly curved plates are coincident with those of perfectly flat, prestressed rectangular plates. However, the eigenmodes are very different from those of initially flat prestressed rectangular plates. The total deflection is found to be the sum of the initial deflection, the deflection resulting from the solution of the flat plate problem, and the deflection resulting from the static problem.

Characterization of the Pivotal Carbon Metabolism of Streptococcus suis Serotype 2 under ex Vivo and Chemically Defined in Vitro Conditions by Isotopologue Profiling*

PubMed Central

Willenborg, Jörg; Huber, Claudia; Koczula, Anna; Lange, Birgit; Eisenreich, Wolfgang; Valentin-Weigand, Peter; Goethe, Ralph

2015-01-01

Streptococcus suis is a neglected zoonotic pathogen that has to adapt to the nutritional requirements in the different host niches encountered during infection and establishment of invasive diseases. To dissect the central metabolic activity of S. suis under different conditions of nutrient availability, we performed labeling experiments starting from [13C]glucose specimens and analyzed the resulting isotopologue patterns in amino acids of S. suis grown under in vitro and ex vivo conditions. In combination with classical growth experiments, we found that S. suis is auxotrophic for Arg, Gln/Glu, His, Leu, and Trp in chemically defined medium. De novo biosynthesis was shown for Ala, Asp, Ser, and Thr at high rates and for Gly, Lys, Phe, Tyr, and Val at moderate or low rates, respectively. Glucose degradation occurred mainly by glycolysis and to a minor extent by the pentose phosphate pathway. Furthermore, the exclusive formation of oxaloacetate by phosphoenolpyruvate (PEP) carboxylation became evident from the patterns in de novo synthesized amino acids. Labeling experiments with S. suis grown ex vivo in blood or cerebrospinal fluid reflected the metabolic adaptation to these host niches with different nutrient availability; however, similar key metabolic activities were identified under these conditions. This points at the robustness of the core metabolic pathways in S. suis during the infection process. The crucial role of PEP carboxylation for growth of S. suis in the host was supported by experiments with a PEP carboxylase-deficient mutant strain in blood and cerebrospinal fluid. PMID:25575595

Effects of reproductive status and high ambient temperatures on the body temperature of a free-ranging basoendotherm.

PubMed

Levesque, Danielle L; Lobban, Kerileigh D; Lovegrove, Barry G

2014-12-01

Tenrecs (Order Afrosoricida) exhibit some of the lowest body temperatures (T b) of any eutherian mammal. They also have a high level of variability in both active and resting T bs and, at least in cool temperatures in captivity, frequently employ both short- and long-term torpor. The use of heterothermy by captive animals is, however, generally reduced during gestation and lactation. We present data long-term T b recordings collected from free-ranging S. setosus over the course of two reproductive seasons. In general, reproductive females had slightly higher (~32 °C) and less variable T b, whereas non-reproductive females and males showed both a higher propensity for torpor as well as lower (~30.5 °C) and more variable rest-phase T bs. Torpor expression defined using traditional means (using a threshold or cut-off T b) was much lower than predicted based on the high degree of heterothermy in captive tenrecs. However, torpor defined in this manner is likely to be underestimated in habitats where ambient temperature is close to T b. Our results caution against inferring metabolic states from T b alone and lend support to the recent call to define torpor in free-ranging animals based on mechanistic and not descriptive variables. In addition, lower variability in T b observed during gestation and lactation confirms that homeothermy is essential for reproduction in this species and probably for basoendothermic mammals in general. The relatively low costs of maintaining homeothermy in a sub-tropical environment might help shed light on how homeothermy could have evolved incrementally from an ancestral heterothermic condition.

Validity of the inexpensive Stepping Meter in counting steps in free living conditions: a pilot study

PubMed Central

De Cocker, K; Cardon, G; De Bourdeaudhuij, I

2006-01-01

Objectives To evaluate if inexpensive Stepping Meters are valid in counting steps in adults in free living conditions. Methods For six days, 35 healthy volunteers wore a criterion Yamax Digiwalker and five Stepping Meters every day until all 973 pedometers had been tested. Steps were recorded daily, and the differences between counts from the Digiwalker and the Stepping Meter were expressed as a percentage of the valid value of the Digiwalker step counts. The criterion used to determine if a Stepping Meter was valid was a maximum deviation of 10% from the Digiwalker step counts. Results A total of 252 (25.9%) Stepping Meters met the criterion, whereas 74.1% made an overestimation or underestimation of more than 10%. In more than one third (36.6%) of the invalid Stepping Meters, the deviation was greater than 50%. Most (64.8%) of the invalid pedometers overestimated the actual steps taken. Conclusions Inexpensive Stepping Meters cannot be used in community interventions as they will give participants the wrong message. PMID:16790485

Free will as relative freedom with conscious component.

PubMed

Hájícek, P

2009-03-01

The general notion of relative freedom is introduced. It is a kind of freedom that is observed everywhere in nature. In biology, incomplete knowledge is defined for all organisms. They cope with the problem by Popper's trial-and-error processes. One source of their success is the relative freedom of choice from the basic option ranges: mutations, motions and neuron connections. After the conjecture is adopted that communicability can be used as a criterion of consciousness, free will is defined as a conscious version of relative freedom. The resulting notion is logically self-consistent and it describes an observable phenomenon that agrees with our experience.

Defining conservation targets on a landscape-scale

USGS Publications Warehouse

Benscoter, A.M.; Romañach, Stephanie; Brandt, Laura A.

2015-01-01

Conservation planning, the process of deciding how to protect, conserve, enhance and(or) minimize loss of natural and cultural resources, is a fundamental process to achieve conservation success in a time of rapid environmental change. Conservation targets, the measurable expressions of desired resource conditions, are an important tool in biological planning to achieve effective outcomes. Conservation targets provide a focus for planning, design, conservation action, and collaborative monitoring of environmental trends to guide landscape-scale conservation to improve the quality and quantity of key ecological and cultural resources. It is essential to have an iterative and inclusive method to define conservation targets that is replicable and allows for the evaluation of the effectiveness of conservation targets over time. In this document, we describe a process that can be implemented to achieve landscape-scale conservation, which includes defining conservation targets. We also describe what has been accomplished to date (September 2015) through this process for the Peninsular Florida Landscape Conservation Cooperative (PFLCC).

The Limit of Free Magnetic Energy in Active Regions

NASA Technical Reports Server (NTRS)

Moore, Ron; Falconer, David; Sterling, Alphonse

2012-01-01

By measuring from active-region magnetograms a proxy of the free energy in the active region fs magnetic field, it has been found previously that (1) there is an abrupt upper limit to the free energy the field can hold that increases with the amount of magnetic field in the active region, the active region fs magnetic flux content, and (2) the free energy is usually near its limit when the field explodes in a CME/flare eruption. That is, explosive active regions are concentrated in a main-sequence path bordering the free-energy ]limit line in (flux content, free-energy proxy) phase space. Here, from measurement of Marshall Space Flight Center vector magnetograms, we find the magnetic condition that underlies the free ]energy limit and the accompanying main sequence of explosive active regions. Using a suitable free ]energy proxy measured from vector magnetograms of 44 active regions, we find that (1) in active regions at and near their free ]energy limit, the ratio of magnetic-shear free energy to the non ]free magnetic energy the potential field would have is approximately 1 in the core field, the field rooted along the neutral line, and (2) this ratio is progressively less in active regions progressively farther below their free ]energy limit. This shows that most active regions in which this core-field energy ratio is much less than 1 cannot be triggered to explode; as this ratio approaches 1, most active regions become capable of exploding; and when this ratio is 1 or greater, most active regions are compelled to explode. From these results we surmise the magnetic condition that determines the free ]energy limit is the ratio of the free magnetic energy to the non-free energy the active region fs field would have were it completely relaxed to its potential ]field configuration, and that this ratio is approximately 1 at the free-energy limit and in the main sequence of explosive active regions.

Insertional Mutations in the Hydrogenase vhc and frc Operons Encoding Selenium-Free Hydrogenases in Methanococcus voltae

PubMed Central

Berghofer, Y.; Klein, A.

1995-01-01

Methanococcus voltae, which contains four different gene groups that encode [NiFe]-hydrogenases, was transformed with integration vectors to achieve polar inactivation of two of the four hydrogenase operons that encode the selenium-free enzymes Vhc and Frc. Transformants which were selected by their acquired puromycin resistance showed site-specific insertions in either the vhc or frc operon by single crossover events. Southern hybridization revealed tandem integrations of whole vectors in the vhc operon, whereas only one vector copy was found in the frc operon. Northern (RNA) hybridizations showed a pac transcript of defined size, indicating strong termination in front of the hydrogenase genes downstream. In spite of the apparent abolition of expression of selenium-free hydrogenases through these polar insertions, they were not lethal to cells upon growth in selenium-deprived minimal medium, which we had previously shown to strongly induce transcription of the respective operons in M. voltae. Instead, like wild-type control cultures, transformants responded to selenium deprivation only with a reduction in growth rate. We conclude that loss of the potential to express a selenium-free hydrogenase can nevertheless be balanced by very small amounts of selenium hydrogenases under laboratory conditions in which the hydrogen supply is not likely to be a limiting growth factor. PMID:16535019

Necessary conditions for tumbling in the rotational motion

NASA Astrophysics Data System (ADS)

Carrera, Danny H. Z.; Weber, Hans I.

2012-11-01

The goal of this work is the investigation of the necessary conditions for the possible existence of tumbling in rotational motion of rigid bodies. In a stable spinning satellite, tumbling may occur by sufficient strong action of external impulses, when the conical movement characteristic of the stable attitude is de-characterized. For this purpose a methodology is chosen to simplify the study of rotational motions with great amplitude, for example free bodies in space, allowing an extension of the analysis to non-conservative systems. In the case of a satellite in space, the projection of the angular velocity along the principal axes of inertia must be known, defining completely the initial conditions of motion for stability investigations. In this paper, the coordinate systems are established according to the initial condition in order to allow a simple analytical work on the equations of motion. Also it will be proposed the definition of a parameter, calling it tumbling coefficient, to measure the intensity of the tumbling and the amplitude of the motion when crossing limits of stability in the concept of Lyapunov. Tumbling in the motion of bodies in space is not possible when this coefficient is positive. Magnus Triangle representation will be used to represent the geometry of the body, establishing regions of stability/instability for possible initial conditions of motion. In the study of nonconservative systems for an oblate body, one sufficient condition will be enough to assure damped motion, and this condition is checked for a motion damped by viscous torques. This paper seeks to highlight the physical understanding of the phenomena and the influence of various parameters that are important in the process.

Elastic modelling in tilted transversely isotropic media with convolutional perfectly matched layer boundary conditions

NASA Astrophysics Data System (ADS)

Han, Byeongho; Seol, Soon Jee; Byun, Joongmoo

2012-04-01

To simulate wave propagation in a tilted transversely isotropic (TTI) medium with a tilting symmetry-axis of anisotropy, we develop a 2D elastic forward modelling algorithm. In this algorithm, we use the staggered-grid finite-difference method which has fourth-order accuracy in space and second-order accuracy in time. Since velocity-stress formulations are defined for staggered grids, we include auxiliary grid points in the z-direction to meet the free surface boundary conditions for shear stress. Through comparisons of displacements obtained from our algorithm, not only with analytical solutions but also with finite element solutions, we are able to validate that the free surface conditions operate appropriately and elastic waves propagate correctly. In order to handle the artificial boundary reflections efficiently, we also implement convolutional perfectly matched layer (CPML) absorbing boundaries in our algorithm. The CPML sufficiently attenuates energy at the grazing incidence by modifying the damping profile of the PML boundary. Numerical experiments indicate that the algorithm accurately expresses elastic wave propagation in the TTI medium. At the free surface, the numerical results show good agreement with analytical solutions not only for body waves but also for the Rayleigh wave which has strong amplitude along the surface. In addition, we demonstrate the efficiency of CPML for a homogeneous TI medium and a dipping layered model. Only using 10 grid points to the CPML regions, the artificial reflections are successfully suppressed and the energy of the boundary reflection back into the effective modelling area is significantly decayed.

Defining the Difference: Comparing Integrated and Traditional Single-Subject Lessons

ERIC Educational Resources Information Center

Zhbanova, Ksenia S.; Rule, Audrey C.; Montgomery, Sarah E.; Nielsen, Lynn E.

2010-01-01

Early childhood curricula should be authentic and child-centered, however, many teachers still rely on direct instruction lessons. To better define how an integrated curriculum meets the needs of students, this study examined teacher talk and actions during instructional activities with first and second graders under two conditions: (1)…

Free radicals: properties, sources, targets, and their implication in various diseases.

PubMed

Phaniendra, Alugoju; Jestadi, Dinesh Babu; Periyasamy, Latha

2015-01-01

Free radicals and other oxidants have gained importance in the field of biology due to their central role in various physiological conditions as well as their implication in a diverse range of diseases. The free radicals, both the reactive oxygen species (ROS) and reactive nitrogen species (RNS), are derived from both endogenous sources (mitochondria, peroxisomes, endoplasmic reticulum, phagocytic cells etc.) and exogenous sources (pollution, alcohol, tobacco smoke, heavy metals, transition metals, industrial solvents, pesticides, certain drugs like halothane, paracetamol, and radiation). Free radicals can adversely affect various important classes of biological molecules such as nucleic acids, lipids, and proteins, thereby altering the normal redox status leading to increased oxidative stress. The free radicals induced oxidative stress has been reported to be involved in several diseased conditions such as diabetes mellitus, neurodegenerative disorders (Parkinson's disease-PD, Alzheimer's disease-AD and Multiple sclerosis-MS), cardiovascular diseases (atherosclerosis and hypertension), respiratory diseases (asthma), cataract development, rheumatoid arthritis and in various cancers (colorectal, prostate, breast, lung, bladder cancers). This review deals with chemistry, formation and sources, and molecular targets of free radicals and it provides a brief overview on the pathogenesis of various diseased conditions caused by ROS/RNS.

Recombinant human albumin supports single cell cloning of CHO cells in chemically defined media.

PubMed

Zhu, Jiang; Wooh, Jong Wei; Hou, Jeff Jia Cheng; Hughes, Benjamin S; Gray, Peter P; Munro, Trent P

2012-01-01

Biologic drugs, such as monoclonal antibodies, are commonly made using mammalian cells in culture. The cell lines used for manufacturing should ideally be clonal, meaning derived from a single cell, which represents a technically challenging process. Fetal bovine serum is often used to support low cell density cultures, however, from a regulatory perspective, it is preferable to avoid animal-derived components to increase process consistency and reduce the risk of contamination from adventitious agents. Chinese hamster ovary (CHO) cells are the most widely used cell line in industry and a large number of serum-free, protein-free, and fully chemically defined growth media are commercially available, although these media alone do not readily support efficient single cell cloning. In this work, we have developed a simple, fully defined, single-cell cloning media, specifically for CHO cells, using commercially available reagents. Our results show that a 1:1 mixture of CD-CHO™ and DMEM/F12 supplemented with 1.5 g/L of recombinant albumin (Albucult®) supports single cell cloning. This formulation can support recovery of single cells in 43% of cultures compared to 62% in the presence of serum. Copyright © 2012 American Institute of Chemical Engineers (AIChE).

Serum-free culture of primary human hepatocytes in a miniaturized hollow-fibre membrane bioreactor for pharmacological in vitro studies.

PubMed

Lübberstedt, Marc; Müller-Vieira, Ursula; Biemel, Klaus M; Darnell, Malin; Hoffmann, Stefan A; Knöspel, Fanny; Wönne, Eva C; Knobeloch, Daniel; Nüssler, Andreas K; Gerlach, Jörg C; Andersson, Tommy B; Zeilinger, Katrin

2015-09-01

Primary human hepatocytes represent an important cell source for in vitro investigation of hepatic drug metabolism and disposition. In this study, a multi-compartment capillary membrane-based bioreactor technology for three-dimensional (3D) perfusion culture was further developed and miniaturized to a volume of less than 0.5 ml to reduce demand for cells. The miniaturized bioreactor was composed of two capillary layers, each made of alternately arranged oxygen and medium capillaries serving as a 3D culture for the cells. Metabolic activity and stability of primary human hepatocytes was studied in this bioreactor in the presence of 2.5% fetal calf serum (FCS) under serum-free conditions over a culture period of 10 days. The miniaturized bioreactor showed functions comparable to previously reported data for larger variants. Glucose and lactate metabolism, urea production, albumin synthesis and release of intracellular enzymes (AST, ALT, GLDH) showed no significant differences between serum-free and serum-supplemented bioreactors. Activities of human-relevant cytochrome P450 (CYP) isoenzymes (CYP1A2, CYP3A4/5, CYP2C9, CYP2D6, CYP2B6) analyzed by determination of product formation rates from selective probe substrates were also comparable in both groups. Gene expression analysis showed moderately higher expression in the majority of CYP enzymes, transport proteins and enzymes of Phase II metabolism in the serum-free bioreactors compared to those maintained with FCS. In conclusion, the miniaturized bioreactor maintained stable function over the investigated period and thus provides a suitable system for pharmacological studies on primary human hepatocytes under defined serum-free conditions. Copyright © 2012 John Wiley & Sons, Ltd.

Efficiently Specified Ventral Midbrain Dopamine Neurons from Human Pluripotent Stem Cells Under Xeno‐Free Conditions Restore Motor Deficits in Parkinsonian Rodents

PubMed Central

Gantner, Carlos W.; Alsanie, Walaa F.; McDougall, Stuart J.; Bye, Chris R.; Elefanty, Andrew G.; Stanley, Edouard G.; Haynes, John M.; Pouton, Colin W.; Thompson, Lachlan H.

2016-01-01

Abstract Recent studies have shown evidence for the functional integration of human pluripotent stem cell (hPSC)‐derived ventral midbrain dopamine (vmDA) neurons in animal models of Parkinson’s disease. Although these cells present a sustainable alternative to fetal mesencephalic grafts, a number of hurdles require attention prior to clinical translation. These include the persistent use of xenogeneic reagents and challenges associated with scalability and storage of differentiated cells. In this study, we describe the first fully defined feeder‐ and xenogeneic‐free protocol for the generation of vmDA neurons from hPSCs and utilize two novel reporter knock‐in lines (LMX1A‐eGFP and PITX3‐eGFP) for in‐depth in vitro and in vivo tracking. Across multiple embryonic and induced hPSC lines, this “next generation” protocol consistently increases both the yield and proportion of vmDA neural progenitors (OTX2/FOXA2/LMX1A) and neurons (FOXA2/TH/PITX3) that display classical vmDA metabolic and electrophysiological properties. We identify the mechanism underlying these improvements and demonstrate clinical applicability with the first report of scalability and cryopreservation of bona fide vmDA progenitors at a time amenable to transplantation. Finally, transplantation of xeno‐free vmDA progenitors from LMX1A‐ and PITX3‐eGFP reporter lines into Parkinsonian rodents demonstrates improved engraftment outcomes and restoration of motor deficits. These findings provide important and necessary advancements for the translation of hPSC‐derived neurons into the clinic. Stem Cells Translational Medicine 2017;6:937–948 PMID:28297587

Guidelines for the analysis of free energy calculations.

PubMed

Klimovich, Pavel V; Shirts, Michael R; Mobley, David L

2015-05-01

Free energy calculations based on molecular dynamics simulations show considerable promise for applications ranging from drug discovery to prediction of physical properties and structure-function studies. But these calculations are still difficult and tedious to analyze, and best practices for analysis are not well defined or propagated. Essentially, each group analyzing these calculations needs to decide how to conduct the analysis and, usually, develop its own analysis tools. Here, we review and recommend best practices for analysis yielding reliable free energies from molecular simulations. Additionally, we provide a Python tool, alchemical-analysis.py, freely available on GitHub as part of the pymbar package (located at http://github.com/choderalab/pymbar), that implements the analysis practices reviewed here for several reference simulation packages, which can be adapted to handle data from other packages. Both this review and the tool covers analysis of alchemical calculations generally, including free energy estimates via both thermodynamic integration and free energy perturbation-based estimators. Our Python tool also handles output from multiple types of free energy calculations, including expanded ensemble and Hamiltonian replica exchange, as well as standard fixed ensemble calculations. We also survey a range of statistical and graphical ways of assessing the quality of the data and free energy estimates, and provide prototypes of these in our tool. We hope this tool and discussion will serve as a foundation for more standardization of and agreement on best practices for analysis of free energy calculations.