Theory and Practice of Lineage Tracing.
Hsu, Ya-Chieh
2015-11-01
Lineage tracing is a method that delineates all progeny produced by a single cell or a group of cells. The possibility of performing lineage tracing initiated the field of Developmental Biology and continues to revolutionize Stem Cell Biology. Here, I introduce the principles behind a successful lineage-tracing experiment. In addition, I summarize and compare different methods for conducting lineage tracing and provide examples of how these strategies can be implemented to answer fundamental questions in development and regeneration. The advantages and limitations of each method are also discussed. © 2015 AlphaMed Press.
Genetic lineage tracing identifies in situ Kit-expressing cardiomyocytes
Liu, Qiaozhen; Yang, Rui; Huang, Xiuzhen; Zhang, Hui; He, Lingjuan; Zhang, Libo; Tian, Xueying; Nie, Yu; Hu, Shengshou; Yan, Yan; Zhang, Li; Qiao, Zengyong; Wang, Qing-Dong; Lui, Kathy O; Zhou, Bin
2016-01-01
Cardiac cells marked by c-Kit or Kit, dubbed cardiac stem cells (CSCs), are in clinical trials to investigate their ability to stimulate cardiac regeneration and repair. These studies were initially motivated by the purported cardiogenic activity of these cells. Recent lineage tracing studies using Kit promoter to drive expression of the inducible Cre recombinase showed that these CSCs had highly limited cardiogenic activity, inadequate to support efficient cardiac repair. Here we reassess the lineage tracing data by investigating the identity of cells immediately after Cre labeling. Our instant lineage tracing approach identifies Kit-expressing cardiomyocytes, which are labeled immediately after tamoxifen induction. In combination with long-term lineage tracing experiments, these data reveal that the large majority of long-term labeled cardiomyocytes are pre-existing Kit-expressing cardiomyocytes rather than cardiomyocytes formed de novo from CSCs. This study presents a new interpretation for the contribution of Kit+ cells to cardiomyocytes and shows that Kit genetic lineage tracing over-estimates the cardiogenic activity of Kit+ CSCs. PMID:26634606
Clonal tracing of Sox9+ liver progenitors in oval cell injury
Tarlow, Branden D.; Finegold, Milton J.; Grompe, Markus
2014-01-01
Proliferating ducts, termed “oval cells”, have long thought to be bipotential, i.e. produce both biliary ducts and hepatocytes during chronic liver injury. The precursor to oval cells is considered to be a facultative liver stem cell (LSC). Recent lineage tracing experiments indicated that the LSC is Sox9+ and can replace the bulk of hepatocyte mass in several settings. However, no clonal relationship between Sox9+ cells and the two epithelial liver lineages was established. We labeled Sox9+ mouse liver cells at low density with a multicolor fluorescent confetti reporter. Organoid formation validated the progenitor activity of the labeled population. Sox9+ cells were traced in multiple oval cell injury models using both histology and FACS. Surprisingly, only rare clones containing both hepatocytes and oval cells were found in any experiment. Quantitative analysis showed that Sox9+ cells contributed only minimally (<1%) to the hepatocyte pool, even in classic oval cell injury models. In contrast, clonally marked mature hepatocytes demonstrated the ability to self-renew in all classic mouse oval cell activation injuries. A hepatocyte chimera model to trace hepatocytes and non-parenchymal cells also demonstrated the prevalence of hepatocyte-driven regeneration in mouse oval cell injury models. Conclusion Sox9+ ductal progenitor cells give rise to clonal oval cell proliferation and bipotential organoids but rarely produce hepatocytes in vivo. Hepatocytes themselves are the predominant source of new parenchyma cells in prototypical mouse models of oval cell activation. PMID:24700457
NASA Astrophysics Data System (ADS)
Moretto, P.; Ortega, R.; Llabador, Y.; Simonoff, M.; Bénard, J.; Moretto, Ph.
1995-09-01
Macro-and Micro-PIXE analysis were applied to study the mechanisms of cellular resistance to cisplatin, a chemotherapeutic agent, widely used nowadays for the treatment of ovarian cancer. Two cultured cell lines, a cisplatin-sensitive and a resistant one, were compared for their trace elements content and platinum accumulation following in vitro exposure to the drug. Bulk analysis revealed significant differences in copper and iron content between the two lines. Subsequent individual cell microanalysis permitted us to characterize the response of the different morphological cell types of the resistant line. This study showed that the metabolism of some trace metals in cisplatin-resistant cells could be affected but the exact relationship with the resistant phenotype remains to be determined. From a technical point of view, this experiment demonstrated that an accurate measurement of trace elements could be derived from nuclear microprobe analysis of individual cell.
Veh, R W
1991-01-02
For double tracing experiments, wheat germ agglutinin (WGA) molecules labeled with two different haptens are desirable. In the present report the suitability of digoxigenylated WGA (DIG-WGA) for retrograde tracing was investigated. For this purpose the new tracer was pressure injected into rat brains and the transported DIG-WGA visualized via its digoxigenyl group with an alkaline phosphatase linked anti DIG antibody in permanently stained sections of high quality. With fixatives containing 2.5% glutaraldehyde only few positive cells were found. However, at milder fixation conditions (4% paraformaldehyde, 0.05% glutaraldehyde 0.2% picric acid, 30 min) retrogradely labeled cells were detected with a sensitivity comparable to tetramethylbenzidine protocols for conventional WGA-HRP (horseradish peroxidase) tracing. Preliminary experiments suggest excellent suitability for double labeling.
Cheeseman, Bevan L.; Zhang, Dongcheng; Binder, Benjamin J.; Newgreen, Donald F.; Landman, Kerry A.
2014-01-01
Cell lineage tracing is a powerful tool for understanding how proliferation and differentiation of individual cells contribute to population behaviour. In the developing enteric nervous system (ENS), enteric neural crest (ENC) cells move and undergo massive population expansion by cell division within self-growing mesenchymal tissue. We show that single ENC cells labelled to follow clonality in the intestine reveal extraordinary and unpredictable variation in number and position of descendant cells, even though ENS development is highly predictable at the population level. We use an agent-based model to simulate ENC colonization and obtain agent lineage tracing data, which we analyse using econometric data analysis tools. In all realizations, a small proportion of identical initial agents accounts for a substantial proportion of the total final agent population. We term these individuals superstars. Their existence is consistent across individual realizations and is robust to changes in model parameters. This inequality of outcome is amplified at elevated proliferation rate. The experiments and model suggest that stochastic competition for resources is an important concept when understanding biological processes which feature high levels of cell proliferation. The results have implications for cell-fate processes in the ENS. PMID:24501272
Activity-Induced Remodeling of Olfactory Bulb Microcircuits Revealed by Monosynaptic Tracing
Arenkiel, Benjamin R.; Hasegawa, Hiroshi; Yi, Jason J.; Larsen, Rylan S.; Wallace, Michael L.; Philpot, Benjamin D.; Wang, Fan; Ehlers, Michael D.
2011-01-01
The continued addition of new neurons to mature olfactory circuits represents a remarkable mode of cellular and structural brain plasticity. However, the anatomical configuration of newly established circuits, the types and numbers of neurons that form new synaptic connections, and the effect of sensory experience on synaptic connectivity in the olfactory bulb remain poorly understood. Using in vivo electroporation and monosynaptic tracing, we show that postnatal-born granule cells form synaptic connections with centrifugal inputs and mitral/tufted cells in the mouse olfactory bulb. In addition, newly born granule cells receive extensive input from local inhibitory short axon cells, a poorly understood cell population. The connectivity of short axon cells shows clustered organization, and their synaptic input onto newborn granule cells dramatically and selectively expands with odor stimulation. Our findings suggest that sensory experience promotes the synaptic integration of new neurons into cell type-specific olfactory circuits. PMID:22216277
LIBS and LIFS for rapid detection of Rb traces in blood
NASA Astrophysics Data System (ADS)
Al-Jeffery, Mohammad O.; Telle, Helmut H.
2002-05-01
Tests that can quickly and efficiently detect traces of illegal performance enhancing drugs are becoming essential. Certain performance enhancing drugs lead to an increase in the count of red blood cells. The proportion of blood made up of red cells is normally around 42 percent. At least 90 percent of Rubidium measured in whole blood is located in the red blood cells. If Rubidium Chloride (RbCl) is given to an athlete around 30 minutes before competing and a sample of their blood (a drop on a filter) was subsequently tested for Rubidium content, the test will give a direct indication of the red blood cell count. In this contribution, we describe an efficient and fast test based on spectroscopic techniques that can be used to detect trace levels of Rubidium. Our experiments employed Rubidium nitride (RbNO3) and trace levels down to 0.3 percent were successfully detected.
NASA Astrophysics Data System (ADS)
Mao, Bing-Yu; Sun, Xiao-Yang; Zhang, Hong-Wei; Zhang, Shi-Cui; Wu, Xian-Han
1997-09-01
Epidermal cells of amphioxus at different developmental stages were investigated by electron microscopy and colloidal carbon tracing experiments. Amphioxus epidermal cells showed different ultrastructural characteristics at larval and adult stages. The epidermal cells at all larval stages studied (24 96 h) had numerous vesicles containing electron dense materials in their apical cytoplasm. In tracing experiments, carbon particles were found in apical vesicles and interoellular spaces. Under scanning electron microscope, many crater-like protrusions were observed on the surface of the cells. These results indicated that amphioxus larval epidermal cells may be capable of endocytosis. The epidermal cells of 3-month and adult amphioxus were obviously secretory ones characterized by well-developed peripheral filaments, a prominent Golgi apparatus and abundant apical secretory vesicles. This study also showed that adult amphioxus body surface mucus contained lectin that could agglutinate human red blood cells. The authors propose that the epidermal cells of amphioxus larva and adult may contribute to the immune defense of the amimal by different means.
Skeletal muscle regeneration in Xenopus tadpoles and zebrafish larvae
2012-01-01
Background Mammals are not able to restore lost appendages, while many amphibians are. One important question about epimorphic regeneration is related to the origin of the new tissues and whether they come from mature cells via dedifferentiation and/or from stem cells. Several studies in urodele amphibians (salamanders) indicate that, after limb or tail amputation, the multinucleated muscle fibres do dedifferentiate by fragmentation and proliferation, thereby contributing to the regenerate. In Xenopus laevis tadpoles, however, it was shown that muscle fibres do not contribute directly to the tail regenerate. We set out to study whether dedifferentiation was present during muscle regeneration of the tadpole limb and zebrafish larval tail, mainly by cell tracing and histological observations. Results Cell tracing and histological observations indicate that zebrafish tail muscle do not dedifferentiate during regeneration. Technical limitations did not allow us to trace tadpole limb cells, nevertheless we observed no signs of dedifferentiation histologically. However, ultrastructural and gene expression analysis of regenerating muscle in tadpole tail revealed an unexpected dedifferentiation phenotype. Further histological studies showed that dedifferentiating tail fibres did not enter the cell cycle and in vivo cell tracing revealed no evidences of muscle fibre fragmentation. In addition, our results indicate that this incomplete dedifferentiation was initiated by the retraction of muscle fibres. Conclusions Our results show that complete skeletal muscle dedifferentiation is less common than expected in lower vertebrates. In addition, the discovery of incomplete dedifferentiation in muscle fibres of the tadpole tail stresses the importance of coupling histological studies with in vivo cell tracing experiments to better understand the regenerative mechanisms. PMID:22369050
Tracing cell lineages in videos of lens-free microscopy.
Rempfler, Markus; Stierle, Valentin; Ditzel, Konstantin; Kumar, Sanjeev; Paulitschke, Philipp; Andres, Bjoern; Menze, Bjoern H
2018-06-05
In vitro experiments with cultured cells are essential for studying their growth and migration pattern and thus, for gaining a better understanding of cancer progression and its treatment. Recent progress in lens-free microscopy (LFM) has rendered it an inexpensive tool for label-free, continuous live cell imaging, yet there is only little work on analysing such time-lapse image sequences. We propose (1) a cell detector for LFM images based on fully convolutional networks and residual learning, and (2) a probabilistic model based on moral lineage tracing that explicitly handles multiple detections and temporal successor hypotheses by clustering and tracking simultaneously. (3) We benchmark our method in terms of detection and tracking scores on a dataset of three annotated sequences of several hours of LFM, where we demonstrate our method to produce high quality lineages. (4) We evaluate its performance on a somewhat more challenging problem: estimating cell lineages from the LFM sequence as would be possible from a corresponding fluorescence microscopy sequence. We present experiments on 16 LFM sequences for which we acquired fluorescence microscopy in parallel and generated annotations from them. Finally, (5) we showcase our methods effectiveness for quantifying cell dynamics in an experiment with skin cancer cells. Copyright © 2018 Elsevier B.V. All rights reserved.
NASA Astrophysics Data System (ADS)
Ghosh, Pratik
1992-01-01
The investigations focussed on in vivo NMR imaging studies of magnetic particles with and within neural cells. NMR imaging methods, both Fourier transform and projection reconstruction, were implemented and new protocols were developed to perform "Neuronal Tracing with Magnetic Labels" on small animal brains. Having performed the preliminary experiments with neuronal tracing, new optimized coils and experimental set-up were devised. A novel gradient coil technology along with new rf-coils were implemented, and optimized for future use with small animals in them. A new magnetic labelling procedure was developed that allowed labelling of billions of cells with ultra -small magnetite particles in a short time. The relationships among the viability of such cells, the amount of label and the contrast in the images were studied as quantitatively as possible. Intracerebral grafting of magnetite labelled fetal rat brain cells made it possible for the first time to attempt monitoring in vivo the survival, differentiation, and possible migration of both host and grafted cells in the host rat brain. This constituted the early steps toward future experiments that may lead to the monitoring of human brain grafts of fetal brain cells. Preliminary experiments with direct injection of horse radish peroxidase-conjugated magnetite particles into neurons, followed by NMR imaging, revealed a possible non-invasive alternative, allowing serial study of the dynamic transport pattern of tracers in single living animals. New gradient coils were built by using parallel solid-conductor ribbon cables that could be wrapped easily and quickly. Rapid rise times provided by these coils allowed implementation of fast imaging methods. Optimized rf-coil circuit development made it possible to understand better the sample-coil properties and the associated trade -offs in cases of small but conducting samples.
The repair of low dose UV light-induced damage to human skin DNA in condition of trace amount Mg 2+
NASA Astrophysics Data System (ADS)
Gao, Fang; Guo, Zhouyi; Zheng, Changchun; Wang, Rui; Liu, Zhiming; Meng, Pei; Zhai, Juan
2008-12-01
Ultraviolet light-induced damage to human skin DNA was widely investigated. The primary mechanism of this damage contributed to form cyclobutane pyrimidine dimmers (CPDs). Although the distribution of UV light-induced CPDs within a defined sequence is similar, the damage in cellular environment which shields the nuclear DNA was higher than that in organism in apparent dose. So we use low UVB light as main study agent. Low dose UV-irradiated HDF-a cells (Human Dermal Fibroblasts-adult cells) which is weaker than epidermic cells were cultured with DMEM at different trace amount of Mg2+ (0mmol/L , 0.1mmol/L , 0.2mmol/L, 0.4mmol/L, 0.8mmol/L, 1.2mmol/L) free-serum DMEM and the repair of DNA strands injured were observed. Treat these cells with DNA strand breaks detection, photoproducts detection and the repair of photoproducts detection. Then quantitate the role of trace amount Mg2+ in repair of UV light-induced damage to human skin. The experiment results indicated that epidermic cells have capability of resistance to UV-radiation at a certain extent. And Mg2+ can regulate the UV-induced damage repair and relative vitality. It can offer a rationale and experiment data to relieve UV light-induced skin disease.
A First Analysis of Metallome Biosignatures of Hyperthermophilic Archaea
Cameron, Vyllinniskii; House, Christopher H.; Brantley, Susan L.
2012-01-01
To date, no experimental data has been reported for the metallome of hyperthermophilic microorganisms although their metal requirements for growth are known to be unique. Here, experiments were conducted to determine (i) cellular trace metal concentrations of the hyperthermophilic Archaea Methanococcus jannaschii and Pyrococcus furiosus, and (ii) a first estimate of the metallome for these hyperthermophilic species via ICP-MS. The metal contents of these cells were compared to parallel experiments using the mesophilic bacterium Escherichia coli grown under aerobic and anaerobic conditions. Fe and Zn were typically the most abundant metals in cells. Metal concentrations for E. coli grown aerobically decreased in the order Fe > Zn > Cu > Mo > Ni > W > Co. In contrast, M. jannaschii and P. furiosus show almost the reverse pattern with elevated Ni, Co, and W concentrations. Of the three organisms, a biosignature is potentially demonstrated for the methanogen M. jannaschii that may, in part, be related to the metallome requirements of methanogenesis. The bioavailability of trace metals more than likely has varied through time. If hyperthermophiles are very ancient, then the trace metal patterns observed here may begin to provide some insights regarding Earth's earliest cells and in turn, early Earth chemistry. PMID:23243390
Flow of essential elements in subcellular fractions during oxidative stress.
Lago, Larissa; Nunes, Emilene A; Vigato, Aryane A; Souza, Vanessa C O; Barbosa, Fernando; Sato, João R; Batista, Bruno L; Cerchiaro, Giselle
2017-02-01
Essential trace elements are commonly found in altered concentrations in the brains of patients with neurodegenerative diseases. Many studies in trace metal determination and quantification are conducted in tissue, cell culture or whole brain. In the present investigation, we determined by ICP-MS Fe, Cu, Zn, Ca, Se, Co, Cr, Mg, and Mn in organelles (mitochondria, nuclei) and whole motor neuron cell cultured in vitro. We performed experiments using two ways to access oxidative stress: cell treatments with H 2 O 2 or Aβ-42 peptide in its oligomeric form. Both treatments caused accumulation of markers of oxidative stress, such as oxidized proteins and lipids, and alteration in DNA. Regarding trace elements, cells treated with H 2 O 2 showed higher levels of Zn and lower levels of Ca in nuclei when compared to control cells with no oxidative treatments. On the other hand, cells treated with Aβ-42 peptide in its oligomeric form showed higher levels of Mg, Ca, Fe and Zn in nuclei when compared to control cells. These differences showed that metal flux in cell organelles during an intrinsic external oxidative condition (H 2 O 2 treatment) are different from an intrinsic external neurodegenerative treatment.
Du, Yan; Qin, Yubo; Li, Zizhen; Yang, Xiuying; Zhang, Jingchang; Westwick, Harrison; Tsai, Eve; Cao, Xudong
2017-12-01
A multifunctional nanobiomaterial has been developed by deliberately combining functions of superparamagnetism, fluorescence, and axonal tracing into one material. Superparamagnetic iron oxide nanoparticles were first synthesized and coated with a silica layer to prevent emission quenching through core-dye interactions; a fluorescent molecule, fluorescein isothiocyanate, was doped inside second layer of silica shell to improve photo-stability and to enable further thiol functionalization. Subsequently, biotinylated dextran amine, a sensitive axonal tracing reagent, was immobilized on the thiol-functionalized nanoparticle surfaces. The resulting nanoparticles were characterized by transmission electron microscopy, dynamic light scattering, X-ray diffraction, X-ray photoelectron spectroscopy, UV-Vis spectroscopy, magnetic resonance imaging and fluorescence confocal microscopy. In vitro cell experiments using both undifferentiated and differentiated Neuro-2a cells showed that the cells were able to take up the nanoparticles intracellularly and that the nanoparticles showed good biocompatibilities. In summary, this new material demonstrated promising performances for both optical and magnetic resonance imaging modalities, suggesting its promising potentials in applications such as in non-invasive imaging, particularly in neuronal tracing.
Smith, J.J.; Hasiotis, S.T.
2008-01-01
This study documents the traces and burrowing behaviors of nymphs of the prairie cicada Cicadetta calliope (Hemiptera: Cicadidae), as observed in neoichnological experiments. Cicada nymphs were collected from the C horizons of sandy Fluvents along the Kansas River east of Lawrence, Kansas. The nymphs appeared to be fifth instars, 13-17 mm long and 6-7 mm wide. Nymphs were placed in plastic enclosures containing layers of colored, moist, very fine-grained sand. They burrowed immediately, excavating air-filled, sediment-enclosed cells between 20 mm and 40 mm long and averaging 9 mm wide. Burrowing was completed in three stages: (1) sediment in the forward portion of the cell was excavated and rolled into a ball with the forelimbs; (2) the nymph turned 180?? using a forward roll, and moved to the back of the cell; and (3) the sediment ball was pushed up against the back wall of the cell and kneaded with the forelimbs into a thin layer. Resulting burrow traces are sinuous and distinctly meniscate and demonstrate that insect larvae construct meniscate, backfilled burrows in well-drained terrestrial settings. Cicadetta calliope nymphs and their traces are excellent analogs for meniscate trace fossils commonly found in late Paleozoic-Cenozoic alluvial deposits and paleosols. Such meniscate trace fossils are useful for interpreting the paleoenvironment and paleohydrogeology of the units in which they are found. In addition, such backfilled burrows can be used to supplement the fossil record of cicada-like hemipterans, currently known only from the latest Permian to the Early Triassic. Copyright ?? 2008, SEPM (Society for Sedimentary Geology).
Laser multipass system with interior cell configuration.
Borysow, Jacek; Kostinski, Alexander; Fink, Manfred
2011-10-20
We ask whether it is possible to restore a multipass system alignment after a gas cell is inserted in the central region. Indeed, it is possible, and we report on a remarkably simple rearrangement of a laser multipass system, composed of two spherical mirrors and a gas cell with flat windows in the middle. For example, for a window of thickness d and refractive index of n, adjusting the mirror separation by ≈2d(1-1/n) is sufficient to preserve the laser beam alignment and tracing. This expression is in agreement with ray-tracing computations and our laboratory experiment. Insofar as our solution corrects for spherical aberrations, it may also find applications in microscopy. © 2011 Optical Society of America
Maliken, Bryan D; Kanisicak, Onur; Karch, Jason; Khalil, Hadi; Fu, Xing; Boyer, Justin G; Prasad, Vikram; Zheng, Yi; Molkentin, Jeffery D
2018-04-17
Background -While c-Kit + adult progenitor cells were initially reported to produce new cardiomyocytes in the heart, recent genetic evidence suggests that such events are exceedingly rare. However, to determine if these rare events represent true de novo cardiomyocyte formation we deleted the necessary cardiogenic transcription factors Gata4 and Gata6 from c-Kit-expressing cardiac progenitor cells (CPCs). Methods - Kit allele-dependent lineage tracing and fusion analysis was performed in mice following simultaneous Gata4 and Gata6 cell-type specific deletion to examine rates of putative de novo cardiomyocyte formation from c-Kit + cells. Bone marrow transplantation experiments were used to define the contribution of Kit allele-derived hematopoietic cells versus Kit lineage-dependent cells endogenous to the heart in contributing to apparent de novo lineage-traced cardiomyocytes. A Tie2 CreERT2 transgene was also used to examine the global impact of Gata4 deletion on the mature cardiac endothelial cell network, which was further evaluated with select angiogenesis assays. Results -Deletion of Gata4 in Kit lineage-derived endothelial cells or in total endothelial cells using the Tie2 CreERT2 transgene, but not from bone morrow cells, resulted in profound endothelial cell expansion, defective endothelial cell differentiation, leukocyte infiltration into the heart and a dramatic increase in Kit allele-dependent lineage-traced cardiomyocytes. However, this increase in labeled cardiomyocytes was an artefact of greater leukocyte-cardiomyocyte cellular fusion due to defective endothelial cell differentiation in the absence of Gata4 Conclusions -Past identification of presumed de novo cardiomyocyte formation in the heart from c-Kit + cells using Kit allele lineage tracing appears to be an artefact of labeled leukocyte fusion with cardiomyocytes. Deletion of Gata4 from c-Kit + endothelial progenitor cells or adult endothelial cells negatively impacted angiogenesis and capillary network integrity.
Nuclear microprobe imaging of gallium nitrate in cancer cells
NASA Astrophysics Data System (ADS)
Ortega, Richard; Suda, Asami; Devès, Guillaume
2003-09-01
Gallium nitrate is used in clinical oncology as treatment for hypercalcemia and for cancer that has spread to the bone. Its mechanism of antitumor action has not been fully elucidated yet. The knowledge of the intracellular distribution of anticancer drugs is of particular interest in oncology to better understand their cellular pharmacology. In addition, most metal-based anticancer compounds interact with endogenous trace elements in cells, altering their metabolism. The purpose of this experiment was to examine, by use of nuclear microprobe analysis, the cellular distribution of gallium and endogenous trace elements within cancer cells exposed to gallium nitrate. In a majority of cellular analyses, gallium was found homogeneously distributed in cells following the distribution of carbon. In a smaller number of cells, however, gallium appeared concentrated together with P, Ca and Fe within round structures of about 2-5 μm diameter located in the perinuclear region. These intracellular structures are typical of lysosomial material.
Fabrication of glass gas cells for the HALOE and MAPS satellite experiments
NASA Technical Reports Server (NTRS)
Sullivan, E. M.; Walthall, H. G.
1984-01-01
The Halogen Occultation Experiment (HALOE) and the Measurement of Air Pollution from Satellites (MAPS) experiment are satellite-borne experiments which measure trace constituents in the Earth's atmosphere. The instruments which obtain the data for these experiments are based on the gas filter correlation radiometer measurement technique. In this technique, small samples of the gases of interest are encapsulated in glass cylinders, called gas cells, which act as very selective optical filters. This report describes the techniques employed in the fabrication of the gas cells for the HALOE and MAPS instruments. Details of the method used to fuse the sapphire windows (required for IR transmission) to the glass cell bodies are presented along with detailed descriptions of the jigs and fixtures used during the assembly process. The techniques and equipment used for window inspection and for pairing the HALOE windows are discussed. Cell body materials and the steps involved in preparing the cell bodies for the glass-to-sapphire fusion process are given.
Simulation of photons from plasmas for the applications to display devices
NASA Astrophysics Data System (ADS)
Lee, Hae June; Yoon, Hyun Jin; Lee, Jae Koo
2007-07-01
Numerical modeling of the photon transport of the ultraviolet (UV) and the visible lights are presented for plasma based display devices. The transport of UV lights which undergo resonance trapping by ground state atoms is solved by using the Holstein equation. After the UV lights are transformed to visible lights at the phosphor surfaces, the visible lights experience complicated traces inside the cell and finally are emitted toward the viewing window after having some power loss within the cell. A three-dimensional ray trace of the visible lights is calculated with a radiosity model. These simulations for the photons strengthen plasma discharge modeling for the application to display devices.
ADAM10 Regulates Notch Function in Intestinal Stem Cells of Mice
Tsai, Yu-Hwai; VanDussen, Kelli L.; Sawey, Eric T.; Wade, Alex W.; Kasper, Chelsea; Rakshit, Sabita; Bhatt, Riha G.; Stoeck, Alex; Maillard, Ivan; Crawford, Howard C.; Samuelson, Linda C.; Dempsey, Peter J.
2014-01-01
BACKGROUND & AIMS ADAM10 is a cell surface sheddase that regulates physiological processes including Notch signaling. ADAM10 is expressed in all intestinal epithelial cell types but the requirement for ADAM10 signaling in crypt homeostasis is not well defined. METHODS We analyzed intestinal tissues from mice with constitutive (Vil-Cre;Adam10f/f mice) and conditional (Vil-CreER;Adam10f/f and Lgr5-CreER;Adam10f/f mice) deletion of ADAM10. We performed cell lineage tracing experiments in mice that expressed a gain-of-function allele of Notch in the intestine (Rosa26NICD) or mice with intestine-specific disruption of Notch (Rosa26DN-MAML), to examine the effects of ADAM10 deletion on cell fate specification and intestinal stem cell maintenance. RESULTS Loss of ADAM10 from developing and adult intestine caused lethality associated with altered intestinal morphology, reduced progenitor cell proliferation, and increased secretory cell differentiation. ADAM10 deletion led to the replacement of intestinal cell progenitors with 2 distinct, post-mitotic, secretory cell lineages: intermediate-like (Paneth/goblet) and enteroendocrine cells. Based on analysis of Rosa26NICD and Rosa26DN-MAML mice, we determined that ADAM10 controls these cell fate decisions by regulating Notch signaling. Cell lineage tracing experiments showed that ADAM10 is required for survival of Lgr5+ crypt-based columnar cells. Our findings indicate that Notch-activated stem cells have a competitive advantage for occupation of the stem cell niche. CONCLUSIONS ADAM10 acts in a cell autonomous manner within the intestinal crypt compartment to regulate Notch signaling. This process is required for progenitor cell lineage specification and crypt-based columnar cell maintenance. PMID:25038433
Heterogeneity and phenotypic plasticity of glial cells in the mammalian enteric nervous system.
Boesmans, Werend; Lasrado, Reena; Vanden Berghe, Pieter; Pachnis, Vassilis
2015-02-01
Enteric glial cells are vital for the autonomic control of gastrointestinal homeostasis by the enteric nervous system. Several different functions have been assigned to enteric glial cells but whether these are performed by specialized subtypes with a distinctive phenotype and function remains elusive. We used Mosaic Analysis with Double Markers and inducible lineage tracing to characterize the morphology and dynamic molecular marker expression of enteric GLIA in the myenteric plexus. Functional analysis in individually identified enteric glia was performed by Ca(2+) imaging. Our experiments have identified four morphologically distinct subpopulations of enteric glia in the gastrointestinal tract of adult mice. Marker expression analysis showed that the majority of glia in the myenteric plexus co-express glial fibrillary acidic protein (GFAP), S100β, and Sox10. However, a considerable fraction (up to 80%) of glia outside the myenteric ganglia, did not label for these markers. Lineage tracing experiments suggest that these alternative combinations of markers reflect dynamic gene regulation rather than lineage restrictions. At the functional level, the three myenteric glia subtypes can be distinguished by their differential response to adenosine triphosphate. Together, our studies reveal extensive heterogeneity and phenotypic plasticity of enteric glial cells and set a framework for further investigations aimed at deciphering their role in digestive function and disease. © 2014 Wiley Periodicals, Inc.
Shuttle of lentiviral vectors via transplanted cells in vivo.
Blömer, U; Gruh, I; Witschel, H; Haverich, A; Martin, U
2005-01-01
Lentiviral vectors have turned out to be an efficient method for stable gene transfer in vitro and in vivo. Not only do fields of application include cell marking and tracing following transplantation in vivo, but also the stable delivery of biological active proteins for gene therapy. A variety of cells, however, need immediate transplantation after preparation, for example, to prevent cell death, differentiation or de-differentiation. Although these cells are usually washed several times following lentiviral transduction, there may be the risk of viral vector shuttle via transplanted cells resulting in undesired in vivo transduction of recipient cells. We investigated whether infectious lentiviral particles are transmitted via ex vivo lentivirally transduced cells. To this end, we explored potential viral shuttle via ex vivo lentivirally transduced cardiomyocytes in vitro and following transplantation into the brain and peripheral muscle. We demonstrate that, even after extensive washing, infectious viral vector particles can be detected in cell suspensions. Those lentiviral vector particles were able to transduce target cells in transwell experiments. Moreover, transmitted vector particles stably transduced resident cells of the recipient central nervous system and muscle in vivo. Our results of lentiviral vector shuttle via transduced cardiomyocytes are significant for both ex vivo gene therapy and for lentiviral cell tracing, in particular for investigation of stem cell differentiation in transplantation models and co-cultivation systems.
Ivanina, Anna V; Beniash, Elia; Etzkorn, Markus; Meyers, Tiffany B; Ringwood, Amy H; Sokolova, Inna M
2013-09-15
Estuarine and coastal habitats experience large fluctuations of environmental factors such as temperature, salinity, partial pressure of CO2 ( [Formula: see text] ) and pH; they also serve as the natural sinks for trace metals. Benthic filter-feeding organisms such as bivalves are exposed to the elevated concentrations of metals in estuarine water and sediments that can strongly affect their physiology. The effects of metals on estuarine organisms may be exacerbated by other environmental factors. Thus, a decrease in pH caused by high [Formula: see text] (hypercapnia) can modulate the effects of trace metals by affecting metal bioavailability, accumulation or binding. To better understand the cellular mechanisms of interactions between [Formula: see text] and trace metals in marine bivalves, we exposed isolated mantle cells of the hard clams (Mercenaria mercenaria) to different levels of [Formula: see text] (0.05, 1.52 and 3.01 kPa) and two major trace metal pollutants - cadmium (Cd) and copper (Cu). Elevated [Formula: see text] resulted in a decrease in intracellular pH (pHi) of the isolated mantle cells from 7.8 to 7.4. Elevated [Formula: see text] significantly but differently affected the trace metal accumulation by the cells. Cd uptake was suppressed at elevated [Formula: see text] levels while Cu accumulation has greatly accelerated under hypercapnic conditions. Interestingly, at higher extracellular Cd levels, labile intracellular Cd(2+) concentration remained the same, while intracellular levels of free Zn(2+) increased suggesting that Cd(2+) substitutes bound Zn(2+) in these cells. In contrast, Cu exposure did not affect intracellular Zn(2+) but led to a profound increase in the intracellular levels of labile Cu(2+) and Fe(2+). An increase in the extracellular concentrations of Cd and Cu led to the elevated production of reactive oxygen species under the normocapnic conditions (0.05 kPa [Formula: see text] ); surprisingly, this effect was mitigated in hypercapnia (1.52 and 3.01 kPa). Overall, our data reveal complex and metal-specific interactions between the cellular effects of trace metals and [Formula: see text] in clams and indicate that variations in environmental [Formula: see text] may modulate the biological effects of trace metals in marine organisms. Copyright © 2013 Elsevier B.V. All rights reserved.
McCann, Matthew R; Tamplin, Owen J; Rossant, Janet; Séguin, Cheryle A
2012-01-01
Back pain related to intervertebral disc degeneration is the most common musculoskeletal problem, with a lifetime prevalence of 82%. The lack of effective treatment for this widespread problem is directly related to our limited understanding of disc development, maintenance and degeneration. The aim of this study was to determine the developmental origins of nucleus pulposus cells within the intervertebral disc using a novel notochord-specific Cre mouse. To trace the fate of notochordal cells within the intervertebral disc, we derived a notochord-specific Cre mouse line by targeting the homeobox gene Noto. Expression of this gene is restricted to the node and the posterior notochord during gastrulation [embryonic day 7.5 (E7.5)-E12.5]. The Noto-cre mice were crossed with a conditional lacZ reporter for visualization of notochord fate in whole-mount embryos. We performed lineage-tracing experiments to examine the contribution of the notochord to spinal development from E12.5 through to skeletally mature mice (9 months). Fate mapping studies demonstrated that, following elongation and formation of the primitive axial skeleton, the notochord gives rise to the nucleus pulposus in fully formed intervertebral discs. Cellular localization of β-galactosidase (encoded by lacZ) and cytokeratin-8 demonstrated that both notochordal cells and chondrocyte-like nucleus pulposus cells are derived from the embryonic notochord. These studies establish conclusively that notochordal cells act as embryonic precursors to all cells found within the nucleus pulposus of the mature intervertebral disc. This suggests that notochordal cells might serve as tissue-specific progenitor cells within the disc and establishes the Noto-cre mouse as a unique tool to interrogate the contribution of notochordal cells to both intervertebral disc development and disc degeneration.
McCann, Matthew R.; Tamplin, Owen J.; Rossant, Janet; Séguin, Cheryle A.
2012-01-01
SUMMARY Back pain related to intervertebral disc degeneration is the most common musculoskeletal problem, with a lifetime prevalence of 82%. The lack of effective treatment for this widespread problem is directly related to our limited understanding of disc development, maintenance and degeneration. The aim of this study was to determine the developmental origins of nucleus pulposus cells within the intervertebral disc using a novel notochord-specific Cre mouse. To trace the fate of notochordal cells within the intervertebral disc, we derived a notochord-specific Cre mouse line by targeting the homeobox gene Noto. Expression of this gene is restricted to the node and the posterior notochord during gastrulation [embryonic day 7.5 (E7.5)-E12.5]. The Noto-cre mice were crossed with a conditional lacZ reporter for visualization of notochord fate in whole-mount embryos. We performed lineage-tracing experiments to examine the contribution of the notochord to spinal development from E12.5 through to skeletally mature mice (9 months). Fate mapping studies demonstrated that, following elongation and formation of the primitive axial skeleton, the notochord gives rise to the nucleus pulposus in fully formed intervertebral discs. Cellular localization of β-galactosidase (encoded by lacZ) and cytokeratin-8 demonstrated that both notochordal cells and chondrocyte-like nucleus pulposus cells are derived from the embryonic notochord. These studies establish conclusively that notochordal cells act as embryonic precursors to all cells found within the nucleus pulposus of the mature intervertebral disc. This suggests that notochordal cells might serve as tissue-specific progenitor cells within the disc and establishes the Noto-cre mouse as a unique tool to interrogate the contribution of notochordal cells to both intervertebral disc development and disc degeneration. PMID:22028328
Nuclear microscopy in trace-element biology — from cellular studies to the clinic
NASA Astrophysics Data System (ADS)
Lindh, Ulf
1993-05-01
The concentration and distribution of trace and major elements in cells are of great interest in cell biology. PIXE can provide elemental concentrations in the bulk of cells or organelles as other bulk techniques such as atomic absorption spectrophotometry and nuclear activation analysis. Supplementary information, perhaps more exciting, on the intracellular distributions of trace elements can be provided using nuclear microscopy. Intracellular distributions of trace elements in normal and malignant cells are presented. The toxicity of mercury and cadmium can be prevented by supplementation of the essential trace element selenium. Some results from an experimental animal model are discussed. The intercellular distribution of major and trace elements in isolated blood cells, as revealed by nuclear microscopy, provides useful clinical information. Examples are given concerning inflammatory connective-tissue diseases and the chronic fatigue syndrome.
ADAM10 regulates Notch function in intestinal stem cells of mice.
Tsai, Yu-Hwai; VanDussen, Kelli L; Sawey, Eric T; Wade, Alex W; Kasper, Chelsea; Rakshit, Sabita; Bhatt, Riha G; Stoeck, Alex; Maillard, Ivan; Crawford, Howard C; Samuelson, Linda C; Dempsey, Peter J
2014-10-01
A disintegrin and metalloproteinase domain-containing protein 10 (ADAM10) is a cell surface sheddase that regulates physiologic processes, including Notch signaling. ADAM10 is expressed in all intestinal epithelial cell types, but the requirement for ADAM10 signaling in crypt homeostasis is not well defined. We analyzed intestinal tissues from mice with constitutive (Vil-Cre;Adam10(f/f) mice) and conditional (Vil-CreER;Adam10(f/f) and Leucine-rich repeat-containing GPCR5 [Lgr5]-CreER;Adam10(f/f) mice) deletion of ADAM10. We performed cell lineage-tracing experiments in mice that expressed a gain-of-function allele of Notch in the intestine (Rosa26(NICD)), or mice with intestine-specific disruption of Notch (Rosa26(DN-MAML)), to examine the effects of ADAM10 deletion on cell fate specification and intestinal stem cell maintenance. Loss of ADAM10 from developing and adult intestine caused lethality associated with altered intestinal morphology, reduced progenitor cell proliferation, and increased secretory cell differentiation. ADAM10 deletion led to the replacement of intestinal cell progenitors with 2 distinct, post-mitotic, secretory cell lineages: intermediate-like (Paneth/goblet) and enteroendocrine cells. Based on analysis of Rosa26(NICD) and Rosa26(DN-MAML) mice, we determined that ADAM10 controls these cell fate decisions by regulating Notch signaling. Cell lineage-tracing experiments showed that ADAM10 is required for survival of Lgr5(+) crypt-based columnar cells. Our findings indicate that Notch-activated stem cells have a competitive advantage for occupation of the stem cell niche. ADAM10 acts in a cell autonomous manner within the intestinal crypt compartment to regulate Notch signaling. This process is required for progenitor cell lineage specification and crypt-based columnar cell maintenance. Copyright © 2014 AGA Institute. Published by Elsevier Inc. All rights reserved.
van den Berge, M; Ozcanhan, G; Zijlstra, S; Lindenbergh, A; Sijen, T
2016-03-01
Especially when minute evidentiary traces are analysed, background cell material unrelated to the crime may contribute to detectable levels in the genetic analyses. To gain understanding on the composition of human cell material residing on surfaces contributing to background traces, we performed DNA and mRNA profiling on samplings of various items. Samples were selected by considering events contributing to cell material deposits in exemplary activities (e.g. dragging a person by the trouser ankles), and can be grouped as public objects, private samples, transfer-related samples and washing machine experiments. Results show that high DNA yields do not necessarily relate to an increased number of contributors or to the detection of other cell types than skin. Background cellular material may be found on any type of public or private item. When a major contributor can be deduced in DNA profiles from private items, this can be a different person than the owner of the item. Also when a specific activity is performed and the areas of physical contact are analysed, the "perpetrator" does not necessarily represent the major contributor in the STR profile. Washing machine experiments show that transfer and persistence during laundry is limited for DNA and cell type dependent for RNA. Skin conditions such as the presence of sebum or sweat can promote DNA transfer. Results of this study, which encompasses 549 samples, increase our understanding regarding the prevalence of human cell material in background and activity scenarios. Copyright © 2015 Elsevier Ireland Ltd. All rights reserved.
Silencing Inhibits Cre-Mediated Recombination of the Z/AP and Z/EG Reporters in Adult Cells
Long, Michael A.; Rossi, Fabio M. V.
2009-01-01
Background The Cre-loxP system has been used to enable tissue specific activation, inactivation and mutation of many genes in vivo and has thereby greatly facilitated the genetic dissection of several cellular and developmental processes. In such studies, Cre-reporter strains, which carry a Cre-activated marker gene, are frequently utilized to validate the expression profile of Cre transgenes, to act as a surrogate marker for excision of a second allele, and to irreversibly label cells for lineage tracing experiments. Principal Findings We have studied three commonly used Cre-reporter strains, Z/AP, Z/EG and R26R-EYFP and have demonstrated that although each reporter can be reliably activated by Cre during early development, exposure to Cre in adult hematopoietic cells results in a much lower frequency of marker-positive cells in the Z/AP or Z/EG strains than in the R26R-EYFP strain. In marker negative cells derived from the Z/AP and Z/EG strains, the transgenic promoter is methylated and Cre-mediated recombination of the locus is inhibited. Conclusions These results show that the efficiency of Cre-mediated recombination is not only dependent on the genomic context of a given loxP-flanked sequence, but also on stochastic epigenetic mechanisms underlying transgene variegation. Furthermore, our data highlights the potential shortcomings of utilizing the Z/AP and Z/EG reporters as surrogate markers of excision or in lineage tracing experiments. PMID:19415111
Silencing inhibits Cre-mediated recombination of the Z/AP and Z/EG reporters in adult cells.
Long, Michael A; Rossi, Fabio M V
2009-01-01
The Cre-loxP system has been used to enable tissue specific activation, inactivation and mutation of many genes in vivo and has thereby greatly facilitated the genetic dissection of several cellular and developmental processes. In such studies, Cre-reporter strains, which carry a Cre-activated marker gene, are frequently utilized to validate the expression profile of Cre transgenes, to act as a surrogate marker for excision of a second allele, and to irreversibly label cells for lineage tracing experiments. We have studied three commonly used Cre-reporter strains, Z/AP, Z/EG and R26R-EYFP and have demonstrated that although each reporter can be reliably activated by Cre during early development, exposure to Cre in adult hematopoietic cells results in a much lower frequency of marker-positive cells in the Z/AP or Z/EG strains than in the R26R-EYFP strain. In marker negative cells derived from the Z/AP and Z/EG strains, the transgenic promoter is methylated and Cre-mediated recombination of the locus is inhibited. These results show that the efficiency of Cre-mediated recombination is not only dependent on the genomic context of a given loxP-flanked sequence, but also on stochastic epigenetic mechanisms underlying transgene variegation. Furthermore, our data highlights the potential shortcomings of utilizing the Z/AP and Z/EG reporters as surrogate markers of excision or in lineage tracing experiments.
Detection of trace cobalt ions in in vivo plant cells using a voltammetric interlocking system.
Ly, Suw Young; Shin, Myoung Ho; Lee, Chang Hyun; Lee, Jin Hui; Kim, Mi Sook; Ji, Sang Woo; Park, Dong Won
2013-01-01
This experiment was conducted to establish a system for detecting trace cobalt ions in water and plant tissues using a voltammetric in vivo sensor. Cyclic and stripping voltammetry was devised from hand-made, macro-type implantable three-electrode systems. The results reached micro and nano working ranges at 100 sec accumulation time. The statistical detection limit (S/N) was attained at 6.0 ng L(-1). For the in vivo application, direct assay of cobalt ions was carried out in Eichhornia crassipes (EC) deep tissue in real time with a preconcentration time of 100 s. Interfaced techniques can be interlocked with other control systems.
USDA-ARS?s Scientific Manuscript database
Cations, such as Ca and Mg, are generally thought to alleviate toxicities of trace metals through site-specific competition (as incorporated in the biotic ligand model, BLM). Short term (48 h) experiments were conducted using cowpea (Vigna unguiculata L. Walp.) seedlings in simple nutrient solution...
Yamaguchi, Sonoko; Miura, Chiemi; Ito, Aki; Agusa, Tetsuro; Iwata, Hisato; Tanabe, Shinsuke; Tuyen, Bui Cach; Miura, Takeshi
2007-06-05
To estimate the influence of water contaminants on fish reproduction in the Mekong Delta area, we sampled cultivated male catfish (Pangasianodon hypophthalmus), investigated testicular development, and measured persistent organic pollutants (POPs) and trace element levels in muscle and liver, respectively. Various testes sizes were observed although sampling took place during a short period. Histological analysis revealed that all developmental stages of germ cells were observed in catfish with large testis, whereas only necrotic spermatogonia but no other germ cells were observed in catfish with small testis. In small testis, furthermore, vacuolization and hypertrophy of Sertoli cells were observed. Measurement of POPs in muscle and trace elements in liver demonstrated that there were negative correlations between GSI and the concentrations of Pb, Mo, Rb and As. To clarify possible direct effects of Pb, Mo, Rb and As on spermatogenesis in fish, we investigated the effects of these trace elements on spermatogenesis using in vitro testicular organ culture of Japanese eel (Anguilla japonica). Treatment with each of the trace elements alone did not affect spermatogenesis. However, treatment with 10(-7)M of Pb, 10(-5) and 10(-4)M of Mo, 10(-5)-10(-3)M of Rb or 10(-5)M of As inhibited the spermatogenesis induced by 11-ketotestosterone (11KT). Furthermore, treatment with 10(-4)M of As in combination with 11KT caused necrosis of testicular fragments. Taken together, these results are consistent with the hypothesis that Pb, Mo, Rb and As can exert inhibitory effects on spermatogenesis in catfish inhabiting the Mekong Delta area.
Ansari, Amir Mehdi; Ahmed, A Karim; Matsangos, Aerielle E; Lay, Frank; Born, Louis J; Marti, Guy; Harmon, John W; Sun, Zhaoli
2016-10-01
Green Fluorescent protein (GFP), used as a cellular tag, provides researchers with a valuable method of measuring gene expression and cell tracking. However, there is evidence to suggest that the immunogenicity and cytotoxicity of GFP potentially confounds the interpretation of in vivo experimental data. Studies have shown that GFP expression can deteriorate over time as GFP tagged cells are prone to death. Therefore, the cells that were originally marked with GFP do not survive and cannot be accurately traced over time. This review will present current evidence for the immunogenicity and cytotoxicity of GFP in in vivo studies by characterizing these responses.
Epiphytic cryptogams as a source of bioaerosols and trace gases
NASA Astrophysics Data System (ADS)
Ruckteschler, Nina; Hrabe de Angelis, Isabella; Zartman, Charles E.; Araùjo, Alessandro; Pöschl, Ulrich; Manzi, Antonio O.; Andreae, Meinrat O.; Pöhlker, Christopher; Weber, Bettina
2016-04-01
Cryptogamic covers comprise (cyano-)bacteria, algae, lichens, bryophytes, fungi, and archaea in varying proportions. These organisms do not form flowers, but reproduce by spores or cell cleavage with these reproductive units being dispersed via the atmosphere. As so-called poikilohydric organisms they are unable to regulate their water content, and their physiological activity pattern mainly follows the external water conditions. We hypothesize, that both spore dispersal and the release of trace gases are governed by the moisture patterns of these organisms and thus they could have a greater impact on the atmosphere than previously thought. In order to test this hypothesis, we initiated experiments at the study site Amazonian Tall Tower Observatory (ATTO) in September 2014. We installed microclimate sensors in epiphytic cryptogams at four different heights of a tree to monitor the activity patterns of these organisms. Self-developed moisture probes are used to analyze the water status of the organisms accompanied by light and temperature sensors. The continuously logged data are linked to ongoing measurements of trace gases and particulate bioaerosols to analyze these for the relevance of cryptogams. Here, we are particularly interested in diurnal cycles of coarse mode particles and the atmospheric abundance of fine potassium-rich particles from a currently unknown biogenic source. Based upon the results of this field study we also investigate the bioaerosol and trace gas release patterns of cryptogamic covers under controlled conditions. With this combined approach of field and laboratory experiments we aim to disclose the role of cryptogamic covers in bioaerosol and trace gas release patterns in the Amazonian rainforest.
Houbracken, Isabelle; Bouwens, Luc
2010-01-01
Adult stem cell research has drawn a lot of attention by many researchers, due to its medical hope of cell replacement or regenerative therapy for diabetes patients. Despite the many research efforts to date, there is no consensus on the existence of stem cells in adult pancreas. Genetic lineage tracing experiments have put into serious doubt whether β-cell neogenesis from stem/progenitor cells takes place postnatally. Different in vitro experiments have suggested centroacinar, ductal, acinar, stellate, or yet unidentified clonigenic cells as candidate β-cell progenitors. As in the rest of the adult stem cell field, sound and promising observations have been made. However, these observations still need to be replicated. As an alternative to committed stem/progenitor cells in the pancreas, transdifferentiation or lineage reprogramming of exocrine acinar and endocrine α-cells may be used to generate new β-cells. At present, it is unclear which approach is most medically promising. This article highlights the progress being made in knowledge about tissue stem cells, their existence and availability for therapy in diabetes. Particular attention is given to the assessment of methods to verify the existence of tissue stem cells.
NASA Astrophysics Data System (ADS)
Li, Kai; Qin, Wei; Ding, Dan; Tomczak, Nikodem; Geng, Junlong; Liu, Rongrong; Liu, Jianzhao; Zhang, Xinhai; Liu, Hongwei; Liu, Bin; Tang, Ben Zhong
2013-01-01
Long-term noninvasive cell tracing by fluorescent probes is of great importance to life science and biomedical engineering. For example, understanding genesis, development, invasion and metastasis of cancerous cells and monitoring tissue regeneration after stem cell transplantation require continual tracing of the biological processes by cytocompatible fluorescent probes over a long period of time. In this work, we successfully developed organic far-red/near-infrared dots with aggregation-induced emission (AIE dots) and demonstrated their utilities as long-term cell trackers. The high emission efficiency, large absorptivity, excellent biocompatibility, and strong photobleaching resistance of the AIE dots functionalized by cell penetrating peptides derived from transactivator of transcription proteins ensured outstanding long-term noninvasive in vitro and in vivo cell tracing. The organic AIE dots outperform their counterparts of inorganic quantum dots, opening a new avenue in the development of fluorescent probes for following biological processes such as carcinogenesis.
Universality of clone dynamics during tissue development
NASA Astrophysics Data System (ADS)
Rulands, Steffen; Lescroart, Fabienne; Chabab, Samira; Hindley, Christopher J.; Prior, Nicole; Sznurkowska, Magdalena K.; Huch, Meritxell; Philpott, Anna; Blanpain, Cedric; Simons, Benjamin D.
2018-05-01
The emergence of complex organs is driven by the coordinated proliferation, migration and differentiation of precursor cells. The fate behaviour of these cells is reflected in the time evolution of their progeny, termed clones, which serve as a key experimental observable. In adult tissues, where cell dynamics is constrained by the condition of homeostasis, clonal tracing studies based on transgenic animal models have advanced our understanding of cell fate behaviour and its dysregulation in disease1,2. But what can be learnt from clonal dynamics in development, where the spatial cohesiveness of clones is impaired by tissue deformations during tissue growth? Drawing on the results of clonal tracing studies, we show that, despite the complexity of organ development, clonal dynamics may converge to a critical state characterized by universal scaling behaviour of clone sizes. By mapping clonal dynamics onto a generalization of the classical theory of aerosols, we elucidate the origin and range of scaling behaviours and show how the identification of universal scaling dependences may allow lineage-specific information to be distilled from experiments. Our study shows the emergence of core concepts of statistical physics in an unexpected context, identifying cellular systems as a laboratory to study non-equilibrium statistical physics.
Labeling single cell for in-vivo study of cell fate mapping and lineage tracing
NASA Astrophysics Data System (ADS)
He, Sicong; Xu, Jin; Wu, Yi; Tian, Ye; Sun, Qiqi; Wen, Zilong; Qu, Jianan Y.
2018-02-01
Cell fate mapping and lineage tracing are significant ways to understanding the developmental origins of biological tissues. It requires labeling individual cells and tracing the development of their progeny. We develop an infrared laser-evoked gene operator heat-shock microscope system to achieve single-cell labeling in zebrafish. With a fluorescent thermometry technique, we measure the temperature increase in zebrafish tissues induced by infrared laser and identify the optimal heat shock conditions for single-cell gene induction in different types of zebrafish cells. We use this technique to study the fate mapping of T lymphocytes and discover the distinct waves of lymphopoiesis during the zebrafish development.
Interactions between prefrontal cortex and cerebellum revealed by trace eyelid conditioning.
Kalmbach, Brian E; Ohyama, Tatsuya; Kreider, Joy C; Riusech, Frank; Mauk, Michael D
2009-01-01
Eyelid conditioning has proven useful for analysis of learning and computation in the cerebellum. Two variants, delay and trace conditioning, differ only by the relative timing of the training stimuli. Despite the subtlety of this difference, trace eyelid conditioning is prevented by lesions of the cerebellum, hippocampus, or medial prefrontal cortex (mPFC), whereas delay eyelid conditioning is prevented by cerebellar lesions and is largely unaffected by forebrain lesions. Here we test whether these lesion results can be explained by two assertions: (1) Cerebellar learning requires temporal overlap between the mossy fiber inputs activated by the tone conditioned stimulus (CS) and the climbing fiber inputs activated by the reinforcing unconditioned stimulus (US), and therefore (2) trace conditioning requires activity that outlasts the presentation of the CS in a subset of mossy fibers separate from those activated directly by the CS. By use of electrical stimulation of mossy fibers as a CS, we show that cerebellar learning during trace eyelid conditioning requires an input that persists during the stimulus-free trace interval. By use of reversible inactivation experiments, we provide evidence that this input arises from the mPFC and arrives at the cerebellum via a previously unidentified site in the pontine nuclei. In light of previous PFC recordings in various species, we suggest that trace eyelid conditioning involves an interaction between the persistent activity of delay cells in mPFC-a putative mechanism of working memory-and motor learning in the cerebellum.
Cells of Origin of Epithelial Ovarian Cancers
2015-09-01
cells in oral squamous cell carcinomas by a novel pathway-based lineage tracing approach in a murine model. ! 13! Specific aims: 1. Determine...SUNDARESAN Lineage tracing and clonal analysis of oral cancer initiating cells The goal of this project is to study cancer stem cells /cancer initiating...whether oral cancer cells genetically marked based on their activities for stem cell -related pathways exhibit cancer stem cell properties in vivo by
Generation Of Functional Insulin-Producing Cells In The Gut By Foxo1 Ablation
Talchai, Chutima; Xuan, Shouhong; Kitamura, Tadahiro; DePinho, Ronald A.; Accili, Domenico
2012-01-01
Restoration of regulated insulin secretion is the ultimate goal of type 1 diabetes therapy. Here we show that, surprisingly, somatic ablation of Foxo1 in Neurog3+ enteroendocrine progenitor cells gives rise to gut insulin-positive cells (Ins+) that express markers of mature β-cells, and secrete bioactive insulin as well as C-peptide in response to glucose and sulfonylureas. Lineage tracing experiments show that gut Ins+ cells arise cell-autonomously from Foxo1-deficient cells. Inducible Foxo1 ablation in adult mice also results in the generation of gut Ins+ cells. Following ablation by the β-cell toxin, streptozotocin, gut Ins+ cells regenerate and produce insulin, reversing hyperglycemia in mice. The data indicate that Neurog3+ enteroendocrine progenitors require active Foxo1 to prevent differentiation into Ins+ cells. Foxo1 ablation in gut epithelium may provide an approach to restore insulin production in type 1 diabetes. PMID:22406641
A Method for Extracting Road Boundary Information from Crowdsourcing Vehicle GPS Trajectories.
Yang, Wei; Ai, Tinghua; Lu, Wei
2018-04-19
Crowdsourcing trajectory data is an important approach for accessing and updating road information. In this paper, we present a novel approach for extracting road boundary information from crowdsourcing vehicle traces based on Delaunay triangulation (DT). First, an optimization and interpolation method is proposed to filter abnormal trace segments from raw global positioning system (GPS) traces and interpolate the optimization segments adaptively to ensure there are enough tracking points. Second, constructing the DT and the Voronoi diagram within interpolated tracking lines to calculate road boundary descriptors using the area of Voronoi cell and the length of triangle edge. Then, the road boundary detection model is established integrating the boundary descriptors and trajectory movement features (e.g., direction) by DT. Third, using the boundary detection model to detect road boundary from the DT constructed by trajectory lines, and a regional growing method based on seed polygons is proposed to extract the road boundary. Experiments were conducted using the GPS traces of taxis in Beijing, China, and the results show that the proposed method is suitable for extracting the road boundary from low-frequency GPS traces, multi-type road structures, and different time intervals. Compared with two existing methods, the automatically extracted boundary information was proved to be of higher quality.
A Method for Extracting Road Boundary Information from Crowdsourcing Vehicle GPS Trajectories
Yang, Wei
2018-01-01
Crowdsourcing trajectory data is an important approach for accessing and updating road information. In this paper, we present a novel approach for extracting road boundary information from crowdsourcing vehicle traces based on Delaunay triangulation (DT). First, an optimization and interpolation method is proposed to filter abnormal trace segments from raw global positioning system (GPS) traces and interpolate the optimization segments adaptively to ensure there are enough tracking points. Second, constructing the DT and the Voronoi diagram within interpolated tracking lines to calculate road boundary descriptors using the area of Voronoi cell and the length of triangle edge. Then, the road boundary detection model is established integrating the boundary descriptors and trajectory movement features (e.g., direction) by DT. Third, using the boundary detection model to detect road boundary from the DT constructed by trajectory lines, and a regional growing method based on seed polygons is proposed to extract the road boundary. Experiments were conducted using the GPS traces of taxis in Beijing, China, and the results show that the proposed method is suitable for extracting the road boundary from low-frequency GPS traces, multi-type road structures, and different time intervals. Compared with two existing methods, the automatically extracted boundary information was proved to be of higher quality. PMID:29671792
Novel utilisation of a circular multi-reflection cell applied to materials ageing experiments
NASA Astrophysics Data System (ADS)
Knox, D. A.; King, A. K.; McNaghten, E. D.; Brooks, S. J.; Martin, P. A.; Pimblott, S. M.
2015-04-01
We report on the novel utilisation of a circular multi-reflection (CMR) cell applied to materials ageing experiments. This enabled trace gas detection within a narrow interfacial region located between two sample materials and remotely interrogated with near-infrared sources combined with fibre-optic coupling. Tunable diode laser absorption spectroscopy was used to detect water vapour and carbon dioxide at wavelengths near 1,358 and 2,004 nm, respectively, with corresponding detection limits of 7 and 1,139 ppm m Hz-0.5. The minimum detectable absorption was estimated to be 2.82 × 10-3 over a 1-s average. In addition, broadband absorption spectroscopy was carried out for the detection of acetic acid, using a super-luminescent light emitting diode centred around 1,430 nm. The 69 cm measurement pathlength was limited by poor manufacturing tolerances of the spherical CMR mirrors and the consequent difficulty of collecting all the cell output light.
Learning and disrupting invariance in visual recognition with a temporal association rule
Isik, Leyla; Leibo, Joel Z.; Poggio, Tomaso
2012-01-01
Learning by temporal association rules such as Foldiak's trace rule is an attractive hypothesis that explains the development of invariance in visual recognition. Consistent with these rules, several recent experiments have shown that invariance can be broken at both the psychophysical and single cell levels. We show (1) that temporal association learning provides appropriate invariance in models of object recognition inspired by the visual cortex, (2) that we can replicate the “invariance disruption” experiments using these models with a temporal association learning rule to develop and maintain invariance, and (3) that despite dramatic single cell effects, a population of cells is very robust to these disruptions. We argue that these models account for the stability of perceptual invariance despite the underlying plasticity of the system, the variability of the visual world and expected noise in the biological mechanisms. PMID:22754523
Hollow Waveguide Gas Sensor for Mid-Infrared Trace Gas Analysis
DOE Office of Scientific and Technical Information (OSTI.GOV)
Kim, S; Young, C; Chan, J
2007-07-12
A hollow waveguide mid-infrared gas sensor operating from 1000 cm{sup -1} to 4000 cm{sup -1} has been developed, optimized, and its performance characterized by combining a FT-IR spectrometer with Ag/Ag-halide hollow core optical fibers. The hollow core waveguide simultaneously serves as a light guide and miniature gas cell. CH{sub 4} was used as test analyte during exponential dilution experiments for accurate determination of the achievable limit of detection (LOD). It is shown that the optimized integration of an optical gas sensor module with FT-IR spectroscopy provides trace sensitivity at the few hundreds of parts-per-billion concentration range (ppb, v/v) for CH{submore » 4}.« less
Differential Effects of the Cannabinoid Agonist WIN55,212-2 on Delay and Trace Eyeblink Conditioning
Steinmetz, Adam B.; Freeman, John H.
2014-01-01
Central cannabinoid-1 receptors (CB1R) play a role in the acquisition of delay eyeblink conditioning but not trace eyeblink conditioning in humans and animals. However, it is not clear why trace conditioning is immune to the effects of cannabinoid receptor compounds. The current study examined the effects of variants of delay and trace conditioning procedures to elucidate the factors that determine the effects of CB1R agonists on eyeblink conditioning. In Experiment 1 rats were administered the cannabinoid agonist WIN55,212-2 during delay, long delay, or trace conditioning. Rats were impaired during delay and long delay but not trace conditioning; the impairment was greater for long delay than delay conditioning. Trace conditioning was further examined in Experiment 2 by manipulating the trace interval and keeping constant the conditioned stimulus (CS) duration. It was found that when the trace interval was 300 ms or less WIN55,212-2 administration impaired the rate of learning. Experiment 3 tested whether the trace interval duration or the relative durations of the CS and trace interval were critical parameters influencing the effects of WIN55,212-2 on eyeblink conditioning. Rats were not impaired with a 100 ms CS, 200 ms trace paradigm but were impaired with a 1000 ms CS, 500 ms trace paradigm, indicating that the duration of the trace interval does not matter but the proportion of the interstimulus interval occupied by the CS relative to the trace period is critical. Taken together the results indicate that cannabinoid agonists affect cerebellar learning the CS is longer than the trace interval. PMID:24128358
van der Lans, Milou; Benito, Cristina; Wagstaff, Laura J.
2017-01-01
There is consensus that, distal to peripheral nerve injury, myelin and Remak cells reorganize to form cellular columns, Bungner's bands, which are indispensable for regeneration. However, knowledge of the structure of these regeneration tracks has not advanced for decades and the structure of the cells that form them, denervated or repair Schwann cells, remains obscure. Furthermore, the origin of these cells from myelin and Remak cells and their ability to give rise to myelin cells after regeneration has not been demonstrated directly, although these conversions are believed to be central to nerve repair. Using genetic lineage-tracing and scanning-block face electron microscopy, we show that injury of sciatic nerves from mice of either sex triggers extensive and unexpected Schwann cell elongation and branching to form long, parallel processes. Repair cells are 2- to 3-fold longer than myelin and Remak cells and 7- to 10-fold longer than immature Schwann cells. Remarkably, when repair cells transit back to myelinating cells, they shorten ∼7-fold to generate the typically short internodes of regenerated nerves. The present experiments define novel morphological transitions in injured nerves and show that repair Schwann cells have a cell-type-specific structure that differentiates them from other cells in the Schwann cell lineage. They also provide the first direct evidence using genetic lineage tracing for two basic assumptions in Schwann cell biology: that myelin and Remak cells generate the elongated cells that build Bungner bands in injured nerves and that such cells can transform to myelin cells after regeneration. SIGNIFICANCE STATEMENT After injury to peripheral nerves, the myelin and Remak Schwann cells distal to the injury site reorganize and modify their properties to form cells that support the survival of injured neurons, promote axon growth, remove myelin-associated growth inhibitors, and guide regenerating axons to their targets. We show that the generation of these repair-supportive Schwann cells involves an extensive cellular elongation and branching, often to form long, parallel processes. This generates a distinctive repair cell morphology that is favorable for the formation of the regeneration tracks that are essential for nerve repair. Remyelination, conversely, involves a striking cell shortening to form the typical short myelin cells of regenerated nerves. We also provide evidence for direct lineage relationships between: (1) repair cells and myelin and Remak cells of uninjured nerves and (2) remyelinating cells in regenerated nerves. PMID:28904214
Gomez-Sanchez, Jose A; Pilch, Kjara S; van der Lans, Milou; Fazal, Shaline V; Benito, Cristina; Wagstaff, Laura J; Mirsky, Rhona; Jessen, Kristjan R
2017-09-13
There is consensus that, distal to peripheral nerve injury, myelin and Remak cells reorganize to form cellular columns, Bungner's bands, which are indispensable for regeneration. However, knowledge of the structure of these regeneration tracks has not advanced for decades and the structure of the cells that form them, denervated or repair Schwann cells, remains obscure. Furthermore, the origin of these cells from myelin and Remak cells and their ability to give rise to myelin cells after regeneration has not been demonstrated directly, although these conversions are believed to be central to nerve repair. Using genetic lineage-tracing and scanning-block face electron microscopy, we show that injury of sciatic nerves from mice of either sex triggers extensive and unexpected Schwann cell elongation and branching to form long, parallel processes. Repair cells are 2- to 3-fold longer than myelin and Remak cells and 7- to 10-fold longer than immature Schwann cells. Remarkably, when repair cells transit back to myelinating cells, they shorten ∼7-fold to generate the typically short internodes of regenerated nerves. The present experiments define novel morphological transitions in injured nerves and show that repair Schwann cells have a cell-type-specific structure that differentiates them from other cells in the Schwann cell lineage. They also provide the first direct evidence using genetic lineage tracing for two basic assumptions in Schwann cell biology: that myelin and Remak cells generate the elongated cells that build Bungner bands in injured nerves and that such cells can transform to myelin cells after regeneration. SIGNIFICANCE STATEMENT After injury to peripheral nerves, the myelin and Remak Schwann cells distal to the injury site reorganize and modify their properties to form cells that support the survival of injured neurons, promote axon growth, remove myelin-associated growth inhibitors, and guide regenerating axons to their targets. We show that the generation of these repair-supportive Schwann cells involves an extensive cellular elongation and branching, often to form long, parallel processes. This generates a distinctive repair cell morphology that is favorable for the formation of the regeneration tracks that are essential for nerve repair. Remyelination, conversely, involves a striking cell shortening to form the typical short myelin cells of regenerated nerves. We also provide evidence for direct lineage relationships between: (1) repair cells and myelin and Remak cells of uninjured nerves and (2) remyelinating cells in regenerated nerves. Copyright © 2017 Gomez-Sanchez et al.
Adult stem cell lineage tracing and deep tissue imaging
Fink, Juergen; Andersson-Rolf, Amanda; Koo, Bon-Kyoung
2015-01-01
Lineage tracing is a widely used method for understanding cellular dynamics in multicellular organisms during processes such as development, adult tissue maintenance, injury repair and tumorigenesis. Advances in tracing or tracking methods, from light microscopy-based live cell tracking to fluorescent label-tracing with two-photon microscopy, together with emerging tissue clearing strategies and intravital imaging approaches have enabled scientists to decipher adult stem and progenitor cell properties in various tissues and in a wide variety of biological processes. Although technical advances have enabled time-controlled genetic labeling and simultaneous live imaging, a number of obstacles still need to be overcome. In this review, we aim to provide an in-depth description of the traditional use of lineage tracing as well as current strategies and upcoming new methods of labeling and imaging. [BMB Reports 2015; 48(12): 655-667] PMID:26634741
Escobar, Carolina; Ansermet, François; Magistretti, Pierre J.
2017-01-01
Experience leaves a trace in the nervous system through plasticity. However, the exact meaning of the mnesic trace is poorly defined in current literature. This article provides a historical review of the term trace in neuroscience and psychoanalysis literature, to highlight two relevant aspects: the diachronic and the semantic dimensions. There has been a general interest in diachrony, or a form of evolution of the trace, but its indissociable semantic dimension remains partially disregarded. Although frequently implied, the diachronic and semantic dimensions of the trace are rarely clearly articulated. We situate this discussion into the classical opposition of syntax, or rules of inscription of the trace in the nervous system, and semantics, or the content of the trace, which takes into consideration the attempt of the human being to build coherence. A general observation is that the study of the term trace follows trends of the thought of the given epoch. This historical analysis also reveals the decay of the idea that the trace is reliable to the experience. From the articulation between neurosciences and psychoanalysis in a historical perspective, this review shows that the trend is to consider trace as a production of the subject, resulting in a permanent rewriting in an attempt to give meaning to the experience. This trend is becoming increasingly evident in light of recent research in neurosciences and psychoanalysis. PMID:28690553
Hunt, Pamela S.; Barnet, Robert C.
2015-01-01
Experience-produced deficits in trace conditioning and context conditioning have been useful tools for examining the role of the hippocampus in learning. It has also been suggested that learning in these tasks is especially vulnerable to neurotoxic effects of alcohol during key developmental periods such as adolescence. In five experiments we systematically examined the presence and source of age-dependent vulnerability to the memory-disrupting effects of acute ethanol in trace conditioning and contextual fear conditioning. In Experiment 1a pre-training ethanol disrupted trace conditioning more strongly in adolescent (postnatal day, PD30-35) than adult rats (PD65-75). In Experiment 1b when pre-training ethanol was accompanied by pre-test ethanol no deficit in trace conditioning was observed in adolescents, suggesting that state-dependent retrieval failure mediated ethanol's disruption of trace conditioning at this age. Experiments 2a and 2b examined the effect of ethanol pretreatment on context conditioning. Here, adult but not adolescent rats were impaired in conditioned freezing to context cues. Experiment 2c explored state-dependency of this effect. Pre-training ethanol continued to disrupt context conditioning in adults even when ethanol was also administered prior to test. Collectively these findings reveal clear age-dependent and task-dependent vulnerabilities in ethanol's disruptive effects on hippocampus-dependent memory. Adolescents were more disrupted by ethanol in trace conditioning than adults, and adults were more disrupted by ethanol in context conditioning than adolescents. We suggest that adolescents may be more susceptible to changes in internal state (state-dependent retrieval failure) than adults and that ethanol disrupted performance in trace and context conditioning through different mechanisms. Relevance of these findings to theories of hippocampus function is discussed. PMID:26192910
A Horizontal Multi-Purpose Microbeam System.
Xu, Y; Randers-Pehrson, G; Marino, S A; Garty, G; Harken, A; Brenner, D J
2018-04-21
A horizontal multi-purpose microbeam system with a single electrostatic quadruplet focusing lens has been developed at the Columbia University Radiological Research Accelerator Facility (RARAF). It is coupled with the RARAF 5.5 MV Singleton accelerator (High Voltage Engineering Europa, the Netherlands) and provides micrometer-size beam for single cell irradiation experiments. It is also used as the primary beam for a neutron microbeam and microPIXE (particle induced x-ray emission) experiment because of its high particle fluence. The optimization of this microbeam has been investigated with ray tracing simulations and the beam spot size has been verified by different measurements.
A horizontal multi-purpose microbeam system
NASA Astrophysics Data System (ADS)
Xu, Y.; Randers-Pehrson, G.; Marino, S. A.; Garty, G.; Harken, A.; Brenner, D. J.
2018-04-01
A horizontal multi-purpose microbeam system with a single electrostatic quadruplet focusing lens has been developed at the Columbia University Radiological Research Accelerator Facility (RARAF). It is coupled with the RARAF 5.5 MV Singleton accelerator (High Voltage Engineering Europa, the Netherlands) and provides micrometer-size beam for single cell irradiation experiments. It is also used as the primary beam for a neutron microbeam and microPIXE (particle induced x-ray emission) experiment because of its high particle fluence. The optimization of this microbeam has been investigated with ray tracing simulations and the beam spot size has been verified by different measurements.
Kristjánsson, Tómas; Thorvaldsson, Tómas Páll; Kristjánsson, Arni
2014-01-01
Previous research involving both unimodal and multimodal studies suggests that single-response change detection is a capacity-free process while a discriminatory up or down identification is capacity-limited. The trace/context model assumes that this reflects different memory strategies rather than inherent differences between identification and detection. To perform such tasks, one of two strategies is used, a sensory trace or a context coding strategy, and if one is blocked, people will automatically use the other. A drawback to most preceding studies is that stimuli are presented at separate locations, creating the possibility of a spatial confound, which invites alternative interpretations of the results. We describe a series of experiments, investigating divided multimodal attention, without the spatial confound. The results challenge the trace/context model. Our critical experiment involved a gap before a change in volume and brightness, which according to the trace/context model blocks the sensory trace strategy, simultaneously with a roaming pedestal, which should block the context coding strategy. The results clearly show that people can use strategies other than sensory trace and context coding in the tasks and conditions of these experiments, necessitating changes to the trace/context model.
Yefimova, Svetlana L; Kurilchenko, Irina Yu; Tkacheva, Tatyana N; Kavok, Nataliya S; Todor, Igor N; Lukianova, Nataliya Yu; Chekhun, Vasyl F; Malyukin, Yuriy V
2014-03-01
We report the Förster resonance energy transfer (FRET)-labeling of liposomal vesicles as an effective approach to study in dynamics the interaction of liposomes with living cells of different types (rat hepatocytes, rat bone marrow, mouse fibroblast-like cells and human breast cancer cells) and cell organelles (hepatocyte nuclei). The in vitro experiments were performed using fluorescent microspectroscopic technique. Two fluorescent dyes (DiO as the energy donor and DiI as an acceptor) were preloaded in lipid bilayers of phosphatidylcholine liposomes that ensures the necessary distance between the dyes for effective FRET. The change in time of the donor and acceptor relative fluorescence intensities was used to visualize and trace the liposome-to-cell interaction. We show that FRET-labeling of liposome vesicles allows one to reveal the differences in efficiency and dynamics of these interactions, which are associated with composition, fluidity, and metabolic activity of cell plasma membranes.
A novel cell culture model as a tool for forensic biology experiments and validations.
Feine, Ilan; Shpitzen, Moshe; Roth, Jonathan; Gafny, Ron
2016-09-01
To improve and advance DNA forensic casework investigation outcomes, extensive field and laboratory experiments are carried out in a broad range of relevant branches, such as touch and trace DNA, secondary DNA transfer and contamination confinement. Moreover, the development of new forensic tools, for example new sampling appliances, by commercial companies requires ongoing validation and assessment by forensic scientists. A frequent challenge in these kinds of experiments and validations is the lack of a stable, reproducible and flexible biological reference material. As a possible solution, we present here a cell culture model based on skin-derived human dermal fibroblasts. Cultured cells were harvested, quantified and dried on glass slides. These slides were used in adhesive tape-lifting experiments and tests of DNA crossover confinement by UV irradiation. The use of this model enabled a simple and concise comparison between four adhesive tapes, as well as a straightforward demonstration of the effect of UV irradiation intensities on DNA quantity and degradation. In conclusion, we believe this model has great potential to serve as an efficient research tool in forensic biology. Copyright © 2016 Elsevier Ireland Ltd. All rights reserved.
Whole organism lineage tracing by combinatorial and cumulative genome editing
McKenna, Aaron; Findlay, Gregory M.; Gagnon, James A.; Horwitz, Marshall S.; Schier, Alexander F.; Shendure, Jay
2016-01-01
Multicellular systems develop from single cells through distinct lineages. However, current lineage tracing approaches scale poorly to whole, complex organisms. Here we use genome editing to progressively introduce and accumulate diverse mutations in a DNA barcode over multiple rounds of cell division. The barcode, an array of CRISPR/Cas9 target sites, marks cells and enables the elucidation of lineage relationships via the patterns of mutations shared between cells. In cell culture and zebrafish, we show that rates and patterns of editing are tunable, and that thousands of lineage-informative barcode alleles can be generated. By sampling hundreds of thousands of cells from individual zebrafish, we find that most cells in adult organs derive from relatively few embryonic progenitors. In future analyses, genome editing of synthetic target arrays for lineage tracing (GESTALT) can be used to generate large-scale maps of cell lineage in multicellular systems for normal development and disease. PMID:27229144
King, Andrew L; Sañudo-Wilhelmy, Sergio A; Leblanc, Karine; Hutchins, David A; Fu, Feixue
2011-08-01
Phytoplankton growth can be limited by numerous inorganic nutrients and organic growth factors. Using the subarctic diatom Attheya sp. in culture studies, we examined how the availability of vitamin B(12) and carbon dioxide partial pressure (pCO(2)) influences growth rate, primary productivity, cellular iron (Fe), cobalt (Co), zinc (Zn) and cadmium (Cd) quotas, and the net use efficiencies (NUEs) of these bioactive trace metals (mol C fixed per mol cellular trace metal per day). Under B(12)-replete conditions, cells grown at high pCO(2) had lower Fe, Zn and Cd quotas, and used those trace metals more efficiently in comparison with cells grown at low pCO(2). At high pCO(2), B(12)-limited cells had ~50% lower specific growth and carbon fixation rates, and used Fe ~15-fold less efficiently, and Zn and Cd ~3-fold less efficiently, in comparison with B(12)-replete cells. The observed higher Fe, Zn and Cd NUE under high pCO(2)/B(12)-replete conditions are consistent with predicted downregulation of carbon-concentrating mechanisms. Co quotas of B(12)-replete cells were ∼5- to 14-fold higher in comparison with B(12)-limited cells, suggesting that >80% of cellular Co of B(12)-limited cells was likely from B(12). Our results demonstrate that CO(2) and vitamin B(12) interactively influence growth, carbon fixation, trace metal requirements and trace metal NUE of this diatom. This suggests the need to consider complex feedback interactions between multiple environmental factors for this biogeochemically critical group of phytoplankton in the last glacial maximum as well as the current and future changing ocean.
Essential trace elements and antioxidant status in relation to severity of HIV in Nigerian patients.
Olaniyi, J A; Arinola, O G
2007-01-01
This study was designed to determine the plasma levels of some antioxidants and trace elements in three severity groups of HIV patients compared with non-HIV-infected controls. The plasma levels of antioxidants (total antioxidant, albumin, bilirubin and uric acid) and trace elements (Mg, Fe, Zn, Mn, Cu, Cr, Cd and Se) were estimated spectrophotometrically in controls and patients with CD4 counts of <200; 200-499 and > or =500 cells/microl. Uric acid and Zn were significantly higher, while vitamin E and all the trace elements (except Zn) were significantly lower in HIV-infected patients compared to healthy controls. The highest level of uric acid was observed in those with CD4 counts of <200 cells/microl. All the trace elements (except Zn) were higher in HIV subjects with a CD4 count of 200-499 cells/microl compared to >500 cells/microl. Only uric acid and Zn showed significant correlation with CD4 count. Based on the results of this study, we recommend routine assessment and appropriate supplementation of antioxidants/trace elements in HIV subjects. This supplementation is hoped to strengthen the immune system and reduce the adverse consequences of HIV- related oxidative stress. Copyright 2007 S. Karger AG, Basel.
NASA Astrophysics Data System (ADS)
Faucher, Giulia; Hoffmann, Linn; Bach, Lennart Thomas; Bottini, Cinzia; Erba, Elisabetta; Riebesell, Ulf
2017-04-01
The Cretaceous witnessed intervals of profound perturbation named "Oceanic Anoxic Events (OAEs)" characterized by volcanic injection of large amounts of CO2, ocean anoxia, eutrophication, and introduction of biologically relevant metals. Some of these extreme events were characterized by size reduction and/or morphological changes of a number of nannofossil species. To detect the cause/s of such changes in the fossil record is challenging. Evidence of a correspondence between intervals of high trace metals concentrations and nannofossil dwarfism may be suggestive for a negative effect of these elements on nannoplankton biocalcification process. In order to verify the hypothesis that anomalously high quantities of essential and/or toxic metals were the cause of coccolith dwarfism, we explored the toxicities of a mixture of trace metals on four living coccolithophores species, namely Emiliania huxleyi, Gephyrocapsa oceanica, Pleurochrysis carterae and Coccolithus pelagicus. The trace metals tested were chosen based upon concentration peaks identified in the geological record and upon known trace metal interaction with living coccolithophores algae. Our results demonstrate a species-specific response to trace metal enrichment in living coccolithophores: E. huxleyi, G. oceanica and C. pelagicus showed a decrease in their growth rate with progressively and exponentially increased trace metal concentrations, while P. carterae is unresponsive to trace metal content. Furthermore, E. huxleyi, G. oceanica and C. pelagicus evidenced a decrease in the cell diameter. Smaller coccoliths were detected in E. huxleyi and C. pelagicus, while coccolith of G. oceanica showed a decrease in size only at the highest trace metal concentrations tested. P. carterae size was unresponsive for changing trace metal concentration. Our results on living coccolithophore algae, demonstrate that elevated trace metal concentrations not only affect growth but also coccolith size and/or weight and that there are large differences between different species. These species-specific differences must be considered before morphological features of coccoliths are used to reconstruct paleo-chemical conditions. Following the laboratory experiment results, elevated trace metal conditions in the past oceans could have caused at least part of the observed morphological changes detected during some Mesozoic OAEs.
Silverstein, Steven M; Jackson, Mitchell A; Goldberg, Damien F; Muñoz, Mauricio
2014-01-01
Purpose To evaluate the efficacy of bromfenac ophthalmic solution 0.07% dosed once daily in achieving zero-to-trace (0–5 cells) anterior chamber cells, following cataract surgery with posterior chamber intraocular lens implantation. Methods The study designed employed two Phase III, double-masked, placebo-controlled, multicenter clinical trials of 440 subjects, randomized to either bromfenac ophthalmic solution 0.07% (n=222) or placebo (n=218). Subjects self-dosed once daily, beginning 1 day before undergoing cataract surgery with intraocular lens implantation (day –1) and again on the day of surgery (day 0) and for 14 days postoperatively. Follow-up was on days 1, 3, 8, and 15. The outcome measures included the percentage of subjects with zero-to-trace anterior chamber cells at each visit, as determined by the percentage of subjects with ≤5 anterior chamber cells, overall anterior chamber cell grades, and summed ocular inflammation score (SOIS) (combined anterior chamber cell and flare scores). Results The proportion of subjects with zero-to-trace anterior chamber cells was significantly higher in the bromfenac 0.07% group compared with the placebo group as early as day 3 (P=0.0007), continued at day 8 (P<0.0001), and through day 15 (P<0.0001). At day 15, 80.2% of subjects in the bromfenac 0.07% group achieved zero-to-trace anterior chamber cells compared with 47.2% of subjects who did so in the placebo group. The overall anterior chamber cell scores were significantly lower in the bromfenac 0.07% group compared with the placebo group at days 3, 8, and 15 (P<0.0001 at each visit). The SOIS were also significantly lower in the bromfenac group compared with the placebo group at days 3, 8, and 15 (P<0.0001 at each visit). Conclusion Bromfenac ophthalmic solution 0.07%, dosed once daily was clinically effective in achieving zero-to-trace anterior chamber cell severity after cataract surgery and was superior to placebo in all anterior chamber cell severity and inflammation outcome measures. PMID:24876763
2014-01-01
Persistent spiking in response to a discrete stimulus is considered to reflect the active maintenance of a memory for that stimulus until a behavioral response is made. This response pattern has been reported in learning paradigms that impose a temporal gap between stimulus presentation and behavioral response, including trace eyeblink conditioning. However, it is unknown whether persistent responses are acquired as a function of learning or simply represent an already existing category of response type. This fundamental question was addressed by recording single-unit activity in the medial prefrontal cortex (mPFC) of rabbits during the initial learning phase of trace eyeblink conditioning. Persistent responses to the tone conditioned stimulus were observed in the mPFC during the very first training sessions. Further analysis revealed that most cells with persistent responses showed this pattern during the very first training trial, before animals had experienced paired training. However, persistent cells showed reliable decreases in response magnitude over the first training session, which were not observed on the second day of training or for sessions in which learning criterion was met. This modification of response magnitude was specific to persistent responses and was not observed for cells showing phasic tone-evoked responses. The data suggest that persistent responses to discrete stimuli do not require learning but that the ongoing robustness of such responses over the course of training is modified as a result of experience. Putative mechanisms for this modification are discussed, including changes in cellular or network properties, neuromodulatory tone, and/or the synaptic efficacy of tone-associated inputs. PMID:25080570
NASA Technical Reports Server (NTRS)
Thaller, Lawrence H.; Quinzio, Michael V.
1997-01-01
The investigation of an aberrant cell voltage during the filling of a large lithium thionyl chloride cell summary is at: an aberrant voltage trace was noted during the review of cell filling data; incident was traced to an interruption during filling; experimentation suggested oxidizable sites within the carbon electrode were responsible for the drop in voltage; the voltage anomaly could be reproduced by interrupting the filling of similar cells; and anomalous voltage dip was not due to a short.
Light ray tracing through a leaf cross section
NASA Technical Reports Server (NTRS)
Kumar, R.; Silva, L. F.
1973-01-01
A light ray, incident at about 5 deg to the normal, is geometrically plotted through the drawing of the cross section of a soybean leaf using Fresnel's equations and Snell's law. The optical mediums of the leaf considered for ray tracing are: air, cell sap, chloroplast, and cell wall. The ray is also drawn through the same leaf cross section with cell wall and air as the only optical mediums. The values of the reflection and transmission found from the ray tracing tests agree closely with the experimental results obtained using a Beckman Dk-2A Spectroreflector.
Holden, A A; Haque, S E; Mayer, K U; Ulrich, A C
2013-08-01
Increased production and expansion of the oil sand industry in Alberta are of great benefit to the economy, but they carry major environmental challenges. The volume of fluid fine tailings requiring storage is 840×10(6) m(3) and growing, making it imperative that we better understand the fate and transport of oil sand process-affected water (OSPW) seepage from these facilities. Accordingly, the current study seeks to characterize both a) the potential for major ion and trace element release, and b) the principal biogeochemical processes involved, as tailing pond OSPW infiltrates into, and interacts with, underlying glacial till sediments prior to reaching down gradient aquifers or surface waters. Objectives were addressed through a series of aqueous and solid phase experiments, including radial diffusion cells, an isotope analysis, X-ray diffraction, and sequential extractions. The diffusion cells were also simulated in a reactive transport framework to elucidate key reaction processes. The experiments indicate that the ingress and interaction of OSPW with the glacial till sediment-pore water system will result in: a mitigation of ingressing Na (retardation), displacement and then limited precipitation of exchangeable Ca and Mg (as carbonates), sulfate reduction and subsequent precipitation of the produced sulfides, as well as biodegradation of organic carbon. High concentrations of ingressing Cl (~375 mg L(-1)) and Na (~575 mg L(-1)) (even though the latter is delayed, or retarded) are expected to migrate through the till and into the underlying sand channel. Trace element mobility was influenced by ion exchange, oxidation-reduction, and mineral phase reactions including reductive dissolution of metal oxyhydroxides - in accordance with previous observations within sandy aquifer settings. Furthermore, although several trace elements showed the potential for release (Al, B, Ba, Cd, Mn, Pb, Si, Sr), large-scale mobilization is not supported. Thus, the present results suggest that in addition to the commonly cited naphthenic acids, remediation of OSPW-impacted groundwater will need to address high concentrations of major ions contributing to salinization. Copyright © 2013 Elsevier B.V. All rights reserved.
Diehl, S.F.; Smith, Kathleen S.; Desborough, G.A.; White, W.W.; Lapakko, K.A.; Goldhaber, Martin B.; Fey, David L.
2003-01-01
To assess the potential impact of metal and acid contamination from mine-waste piles, it is important to identify the mineralogic source of trace metals and their mode of occurrence. Microscopic analysis of mine-waste samples from both hard-rock and coalmine waste samples demonstrate a microstructural control, as well as mineralogic control, on the source and release of trace metals into local water systems. The samples discussed herein show multiple periods of sulfide mineralization with varying concentrations of trace metals. In the first case study, two proprietary hard-rock mine-waste samples exposed to a series of humidity cell tests (which simulate intense chemical weathering conditions) generated acid and released trace metals. Some trace elements of interest were: arsenic (45-120 ppm), copper (60-320 ppm), and zinc (30-2,500 ppm). Untested and humidity cell-exposed samples were studied by X-ray diffraction, scanning electron microscope with energy dispersive X-ray (SEM/EDX), and electron microprobe analysis. Studies of one sample set revealed arsenic-bearing pyrite in early iron- and magnesium-rich carbonate-filled microveins, and iron-, copper-, arsenic-, antimony-bearing sulfides in later crosscutting silica-filled microveins. Post humidity cell tests indicated that the carbonate minerals were removed by leaching in the humidity cells, exposing pyrite to oxidative conditions. However, sulfides in the silica-filled veins were more protected. Therefore, the trace metals contained in the sulfides within the silica-filled microveins may be released to the surface and (or) ground water system more slowly over a greater time period. In the second case study, trace metal-rich pyrite-bearing coals from the Warrior Basin, Alabama were analyzed. Arsenic-bearing pyrite was observed in a late-stage pyrite phase in microfaults and microveins that crosscut earlier arsenic.
NASA Astrophysics Data System (ADS)
Shen, Ling; Shen, Yifeng; Li, Feng
2018-01-01
In pursuit of capturing more wave-guided luminescence for surface-printed bifacial GaAs μ-cells, the pyramid structure has been incorporated with specular back side reflector (BSR) to change the direction of photon propagation. Based on ray tracing model, the calculated photon capturing efficiency of GaAs μ-cells from back side via pyramid, dependent on the parameters of pyramid structure, achieve the largest 1.7× increase for dye absorption peak of 480 nm compared to the case without pyramid. More significantly, the short circuit current in experiment has been improved from original 16.5 mA/cm2 to 23.75 mA/cm2 for the AM 1.5G solar spectrum. Further experiment demonstrates that the optimized pyramid structure enables the integrated luminescent intensity to reach ∼3× increase in a smaller distance of optical transport, which means the advantages in photon capturing efficiency for cells with higher aspect ratio. The calculation further confirms that the cells with higher aspect ratio, among all cells with the same area, realize the higher concentration ratio for the same geometric gain. This provides a guideline for design of cell geometries to guarantee a higher power output in terms of cell modules.
Yang, Jinghui; Chen, Jianjun; Young, James S; Wang, Qiang; Yin, Dengping; Sciammas, Roger; Chong, Anita S
2016-08-01
The dual role of B cells as drivers and suppressors of the immune responses have underscored the need to trace the fate of B cells recognizing donor major histocompatibility complex class I and class II after allograft transplantation. In this study, we used donor class II tetramers to trace the fate of I-E-specific B cells after immunization with BALB/c spleen cells or cardiac transplantation, in naive or sensitized C57BL/6 recipients. We combined this approach with genetic lineage tracing of memory B cells in activation-induced cytidine deaminase regulated Cre transgenic mice crossed to the ROSA26-enhanced yellow fluorescent protein reporter mice to track endogenous I-E-specific memory B cell generation. Immunization with BALB/c splenocytes or heart transplantation induced an expansion and differentiation of I-E-specific B cells into germinal center B cells, whereas BALB/c heart transplantation into sensitized recipients induced the preferential differentiation into antibody-secreting cells. A 10.8-fold increase in the frequency of I-E-specific memory B cells was observed by day 42 postimmunization. Treatment with CTLA4-Ig starting on day 0 or day 7 postimmunization abrogated I-E-specific memory B cell generation and sensitized humoral responses, but not if treatment commenced on day 14. The majority of donor-specific memory B cells are generated between days 7 and 14 postimmunization, thus revealing a flexible timeframe whereby delayed CTLA4-Ig administration can inhibit sensitization and the generation of memory graft-reactive B cells.
A comparison of pecking generated by serial, delay, and trace autoshaping procedures1
Newlin, Robert J.; LoLordo, Vincent M.
1976-01-01
Pigeons were exposed to serial, delay, and trace autoshaping procedures. In Experiment I, all conditioned stimuli (CSs) were changes in illumination of the response key. The number of trials to acquisition of the keypeck increased from serial, to 4-sec delay, 8-sec delay, and 8-sec trace procedures, in that order. In Experiment II, which used a longer intertrial interval, trials to criterion increased from 8-sec delay, to 28-sec delay, 8-sec trace, and 28-sec trace procedures, in that order. In Experiment III, two groups received serial procedures in which the first CS was either a tone or a houselight, and the second was a keylight. The tone group acquired the key peck more rapidly than the houselight group. Early in conditioning in these experiments, and when the conditioned stimulus was a change in the keylight, there was a short latency to the onset of pecking and pecking was directed at the CS. After extensive conditioning, or when the CS was relatively diffuse, pecking still occurred, but had a longer latency and was not reliably directed toward the conditioned stimulus. PMID:16811907
Effect of Impurities on the Freezing Point of Zinc
NASA Astrophysics Data System (ADS)
Sun, Jianping; Rudtsch, Steffen; Niu, Yalu; Zhang, Lin; Wang, Wei; Den, Xiaolong
2017-03-01
The knowledge of the liquidus slope of impurities in fixed-point metal defined by the International Temperature Scale of 1990 is important for the estimation of uncertainties and correction of fixed point with the sum of individual estimates method. Great attentions are paid to the effect of ultra-trace impurities on the freezing point of zinc in the National Institute of Metrology. In the present work, the liquidus slopes of Ga-Zn, Ge-Zn were measured with the slim fixed-point cell developed through the doping experiments, and the temperature characteristics of the phase diagram of Fe-Zn were furthermore investigated. A quasi-adiabatic Zn fixed-point cell was developed with the thermometer well surrounded by the crucible with the pure metal, and the temperature uniformity of less than 20 mK in the region where the metal is located was obtained. The previous doping experiment of Pb-Zn with slim fixed-point cell was checked with quasi-adiabatic Zn fixed-point cell, and the result supports the previous liquidus slope measured with the traditional fixed-point realization.
Whole-organism clone tracing using single-cell sequencing.
Alemany, Anna; Florescu, Maria; Baron, Chloé S; Peterson-Maduro, Josi; van Oudenaarden, Alexander
2018-04-05
Embryonic development is a crucial period in the life of a multicellular organism, during which limited sets of embryonic progenitors produce all cells in the adult body. Determining which fate these progenitors acquire in adult tissues requires the simultaneous measurement of clonal history and cell identity at single-cell resolution, which has been a major challenge. Clonal history has traditionally been investigated by microscopically tracking cells during development, monitoring the heritable expression of genetically encoded fluorescent proteins and, more recently, using next-generation sequencing technologies that exploit somatic mutations, microsatellite instability, transposon tagging, viral barcoding, CRISPR-Cas9 genome editing and Cre-loxP recombination. Single-cell transcriptomics provides a powerful platform for unbiased cell-type classification. Here we present ScarTrace, a single-cell sequencing strategy that enables the simultaneous quantification of clonal history and cell type for thousands of cells obtained from different organs of the adult zebrafish. Using ScarTrace, we show that a small set of multipotent embryonic progenitors generate all haematopoietic cells in the kidney marrow, and that many progenitors produce specific cell types in the eyes and brain. In addition, we study when embryonic progenitors commit to the left or right eye. ScarTrace reveals that epidermal and mesenchymal cells in the caudal fin arise from the same progenitors, and that osteoblast-restricted precursors can produce mesenchymal cells during regeneration. Furthermore, we identify resident immune cells in the fin with a distinct clonal origin from other blood cell types. We envision that similar approaches will have major applications in other experimental systems, in which the matching of embryonic clonal origin to adult cell type will ultimately allow reconstruction of how the adult body is built from a single cell.
Woods, Dori C; Tilly, Jonathan L
2017-01-01
Accruing evidence indicates that production of new oocytes (oogenesis) and their enclosure by somatic cells (folliculogenesis) are processes not limited to the perinatal period in mammals. Endpoints ranging from oocyte counts to genetic lineage tracing and transplantation experiments support a paradigm shift in reproductive biology involving active renewal of oocyte-containing follicles during postnatal life. The recent purification of mitotically active oocyte progenitor cells, termed female germline stem cells (fGSCs) or oogonial stem cells (OSCs), from mouse and human ovaries opens up new avenues for research into the biology and clinical utility of these cells. Here we detail methods for the isolation of mouse and human OSCs from adult ovarian tissue, cultivation of the cells after purification, and characterization of the cells before and after ex vivo expansion. The latter methods include analysis of germ cell–specific markers and in vitro oogenesis, as well as the use of intraovarian transplantation to test the oocyte-forming potential of OSCs in vivo. PMID:23598447
Progenitor cell domains in the developing and adult pancreas
Kopp, Janel L; Dubois, Claire L; Hao, Ergeng; Thorel, Fabrizio; Herrera, Pedro L
2011-01-01
Unlike organs with defined stem cell compartments, such as the intestine, the pancreas has limited capacity to regenerate. The question of whether the adult pancreas harbors facultative stem/progenitor cells has been a prime subject of debate. Cumulative evidence from recent genetic lineage tracing studies, in which specific cell populations were marked and traced in adult mice, suggests that endocrine and acinar cells are no longer generated from progenitors in the adult pancreas. These studies further indicate that adult pancreatic ductal cells are not a source for endocrine cells following pancreatic injury, as previously suggested. Our own studies have shown that adult ductal cells reinitiate expression of some endocrine progenitor markers, including Ngn3, after injury by partial duct ligation (PDL), but that these cells do not undergo endocrine cell differentiation. Here, we present additional evidence that endocrine cells do not arise from ducts following β-cell ablation by streptozotocin or by a diphtheria toxin-expressing transgene or when β-cell ablation is combined with PDL. In this review, we discuss findings from recent lineage tracing studies of embryonic and adult pancreatic ductal cells. Based upon the combined evidence from these studies, we propose that multipotency is associated with a specific transcriptional signature. PMID:21558806
Miskovic, Vladimir; Keil, Andreas
2015-01-01
The visual system is biased towards sensory cues that have been associated with danger or harm through temporal co-occurrence. An outstanding question about conditioning-induced changes in visuocortical processing is the extent to which they are driven primarily by top-down factors such as expectancy or by low-level factors such as the temporal proximity between conditioned stimuli and aversive outcomes. Here, we examined this question using two different differential aversive conditioning experiments: participants learned to associate a particular grating stimulus with an aversive noise that was presented either in close temporal proximity (delay conditioning experiment) or after a prolonged stimulus-free interval (trace conditioning experiment). In both experiments we probed cue-related cortical responses by recording steady-state visual evoked potentials (ssVEPs). Although behavioral ratings indicated that all participants successfully learned to discriminate between the grating patterns that predicted the presence versus absence of the aversive noise, selective amplification of population-level responses in visual cortex for the conditioned danger signal was observed only when the grating and the noise were temporally contiguous. Our findings are in line with notions purporting that changes in the electrocortical response of visual neurons induced by aversive conditioning are a product of Hebbian associations among sensory cell assemblies rather than being driven entirely by expectancy-based, declarative processes. PMID:23398582
The effects of trace element content on pyrite oxidation rates
NASA Astrophysics Data System (ADS)
Gregory, D. D.; Lyons, T.; Cliff, J. B.; Perea, D. E.; Johnson, A.; Romaniello, S. J.; Large, R. R.
2017-12-01
Pyrite acts as both an important source and sink for many different metals and metalloids in the environment, including many that are toxic. Oxidation of pyrite can release these elements while at the same time producing significant amounts of sulfuric acid. Such issues are common in the vicinity of abandoned mines and smelters, but, as pyrite is a common accessory mineral in many different lithologies, significant pyrite oxidation can occur whenever pyritic rocks are exposed to oxygenated water or the atmosphere. Accelerated exposure to oxygen can occur during deforestation, fracking for petroleum, and construction projects. Geochemical models for pyrite oxidation can help us develop strategies to mitigate these deleterious effects. An important component of these models is an accurate pyrite oxidation rate; however, current pyrite oxidation rates have been determined using relatively pure pyrite. Natural pyrite is rarely pure and has a wide range of trace element concentrations that may affect the oxidation rate. Furthermore, the position of trace elements within the mineral lattice can also affect the oxidation rate. For example, elements such as Ni and Co, which substitute into the pyrite lattice, are thought to stabilize the lattice and thus prevent pyrite oxidation. Alternatively, trace elements that are held within inclusions of other minerals could form a galvanic cell with the surrounding pyrite, thus enhancing pyrite oxidation rates. In this study, we present preliminary analyses from three different pyrite oxidation experiments each using natural pyrite with different trace element compositions. These results show that the pyrite with the highest trace element concentration has approximately an order of magnitude higher oxidation rate compared to the lowest trace element sample. To further elucidate the mechanisms, we employed microanalytical techniques to investigate how the trace elements are held within the pyrite. LA-ICPMS was used to determine the variability of trace element content from the pyrite samples. These data were then used to select areas of interest for NanoSIMS analyses, which in turn was used to select areas for TEM and APT. These analyses show that the trace element content of pyrite can be highly variable, which may significantly affect the rate of pyrite oxidation.
Zhang, Lixin; Ju, Xiaofang; Wang, Fa; Guo, Zhiwei; Piao, Shanhua; Teng, Chunbo
2008-04-01
Pancreas is an important mixed gland having both endocrine and exocrine functions, and has been proven regeneration after injury. To explore the cell lineage tracing methods in pancreas in vivo and the regenerate cells source, we used pseudo-type retrovirus to transfect adult mouse pancreas which had been partially pancreatectomized by rubbing the kerf using a cotton stick saturated with retrovirus suspension then injecting 100 microL retrovirus suspension into pancreas, injecting 100 microL retrovirus by caudal vein, or interperitoneally injecting retrovirus respectively. The results showed that the method of rubbing the kerf then injection of retrovirus suspension into pancreas could more effectively mark the pancreatic cells than the caudal vein injection and the intraperitoneal injection did in vivo. Furthermore, this study also found that some acinus cells could accept injury stimulus signals to regenerate through resuming mitosis after pancreatic injury. This study establishes a cell lineage tracing method in pancreas in vivo using retrovirus and offers a clue for gene therapy of pancreatic diseases using retrovirus vectors.
Infrared trace element detection system
Bien, F.; Bernstein, L.S.; Matthew, M.W.
1988-11-15
An infrared trace element detection system includes an optical cell into which the sample fluid to be examined is introduced and removed. Also introduced into the optical cell is a sample beam of infrared radiation in a first wavelength band which is significantly absorbed by the trace element and a second wavelength band which is not significantly absorbed by the trace element for passage through the optical cell through the sample fluid. The output intensities of the sample beam of radiation are selectively detected in the first and second wavelength bands. The intensities of a reference beam of the radiation are similarly detected in the first and second wavelength bands. The sensed output intensity of the sample beam in one of the first and second wavelength bands is normalized with respect to the other and similarly, the intensity of the reference beam of radiation in one of the first and second wavelength bands is normalized with respect to the other. The normalized sample beam intensity and normalized reference beam intensity are then compared to provide a signal from which the amount of trace element in the sample fluid can be determined. 11 figs.
Infrared trace element detection system
Bien, Fritz; Bernstein, Lawrence S.; Matthew, Michael W.
1988-01-01
An infrared trace element detection system including an optical cell into which the sample fluid to be examined is introduced and removed. Also introduced into the optical cell is a sample beam of infrared radiation in a first wavelength band which is significantly absorbed by the trace element and a second wavelength band which is not significantly absorbed by the trace element for passage through the optical cell through the sample fluid. The output intensities of the sample beam of radiation are selectively detected in the first and second wavelength bands. The intensities of a reference beam of the radiation are similarly detected in the first and second wavelength bands. The sensed output intensity of the sample beam in one of the first and second wavelength bands is normalized with respect to the other and similarly, the intensity of the reference beam of radiation in one of the first and second wavelength bands is normalized with respect to the other. The normalized sample beam intensity and normalized reference beam intensity are then compared to provide a signal from which the amount of trace element in the sample fluid can be determined.
Barron, Martin; Zhang, Siyuan
2018-01-01
Abstract Cell types in cell populations change as the condition changes: some cell types die out, new cell types may emerge and surviving cell types evolve to adapt to the new condition. Using single-cell RNA-sequencing data that measure the gene expression of cells before and after the condition change, we propose an algorithm, SparseDC, which identifies cell types, traces their changes across conditions and identifies genes which are marker genes for these changes. By solving a unified optimization problem, SparseDC completes all three tasks simultaneously. SparseDC is highly computationally efficient and demonstrates its accuracy on both simulated and real data. PMID:29140455
Pezze, M A; Marshall, H J; Cassaday, H J
2016-08-01
In an appetitively motivated procedure, we have previously reported that systemic treatment with the dopamine (DA) D1 receptor agonist SKF81297 (0.4 and 0.8 mg/kg) depressed acquisition at a 2 s inter-stimulus-interval (ISI), suitable to detect trace conditioning impairment. However since DA is involved in reinforcement processes, the generality of effects across appetitively- and aversively-motivated trace conditioning procedures cannot be assumed. The present study tested the effects of SKF81297 (0.4 and 0.8 mg/kg) in an established conditioned emotional response (CER) procedure. Trace-dependent conditioning was clearly shown in two experiments: while conditioning was relatively strong at a 3-s ISI, it was attenuated at a 30-s ISI. This was shown after two (Experiment 1) or four (Experiment 2) conditioning trials conducted in - as far as possible - the same CER procedure. Contrary to prediction, in neither experiment was there any indication that trace conditioning was attenuated by treatment with 0.4 or 0.8 mg/kg SKF81297. In the same rats, locomotor activity was significantly enhanced at the 0.8 mg/kg dose of SKF81297. These results suggest that procedural details of the trace conditioning variant in use are an important determinant of the profile of dopaminergic modulation.
Safaei, Zahra; Karimi, Keikhosro; Zamani, Akram
2016-08-30
In this study the effects of phosphate, potassium, yeast extract, and trace metals on the growth of Mucor indicus and chitosan, chitin, and metabolite production by the fungus were investigated. Maximum yield of chitosan (0.32 g/g cell wall) was obtained in a phosphate-free medium. Reversely, cell growth and ethanol formation by the fungus were positively affected in the presence of phosphate. In a phosphate-free medium, the highest chitosan content (0.42 g/g cell wall) and cell growth (0.66 g/g sugar) were obtained at 2.5 g/L of KOH. Potassium concentration had no significant effect on ethanol and glycerol yields. The presence of trace metals significantly increased the chitosan yield at an optimal phosphate and potassium concentration (0.50 g/g cell wall). By contrast, production of ethanol by the fungus was negatively affected (0.33 g/g sugars). A remarkable increase in chitin and decrease in chitosan were observed in the absence of yeast extract and concentrations lower than 2 g/L. The maximum chitosan yield of 51% cell wall was obtained at 5 g/L of yeast extract when the medium contained no phosphate, 2.5 g/L KOH, and 1 mL/L trace metal solution.
A novel tracing method for the segmentation of cell wall networks.
De Vylder, Jonas; Rooms, Filip; Dhondt, Stijn; Inze, Dirk; Philips, Wilfried
2013-01-01
Cell wall networks are a common subject of research in biology, which are important for plant growth analysis, organ studies, etc. In order to automate the detection of individual cells in such cell wall networks, we propose a new segmentation algorithm. The proposed method is a network tracing algorithm, exploiting the prior knowledge of the network structure. The method is applicable on multiple microscopy modalities such as fluorescence, but also for images captured using non invasive microscopes such as differential interference contrast (DIC) microscopes.
Microbial Metabolism in Serpentinite Fluids
NASA Astrophysics Data System (ADS)
Crespo-Medina, M.; Brazelton, W. J.; Twing, K. I.; Kubo, M.; Hoehler, T. M.; Schrenk, M. O.
2013-12-01
Serpentinization is the process in which ultramafic rocks, characteristic of the upper mantle, react with water liberating mantle carbon and reducing power to potenially support chemosynthetic microbial communities. These communities may be important mediators of carbon and energy exchange between the deep Earth and the surface biosphere. Our work focuses on the Coast Range Ophiolite Microbial Observatory (CROMO) in Northern California where subsurface fluids are accessible through a series of wells. Preliminary analyses indicate that the highly basic fluids (pH 9-12) have low microbial diversity, but there is limited knowledge about the metabolic capabilities of these communties. Metagenomic data from similar serpentine environments [1] have identified Betaproteobacteria belonging to the order Burkholderiales and Gram-positive bacteria from the order Clostridiales as key components of the serpentine microbiome. In an effort to better characterize the microbial community, metabolism, and geochemistry at CROMO, fluids from two representative wells (N08B and CSWold) were sampled during recent field campaigns. Geochemical characterization of the fluids includes measurements of dissolved gases (H2, CO, CH4), dissolved inorganic and organic carbon, volatile fatty acids, and nutrients. The wells selected can be differentiated in that N08B had higher pH (10-11), lower dissolved oxygen, and cell counts ranging from 105-106 cells mL-1 of fluid, with an abundance of the betaproteobacterium Hydrogenophaga. In contrast, fluids from CSWold have slightly lower pH (9-9.5), DO, and conductivity, as well as higher TDN and TDP. CSWold fluid is also characterized for having lower cell counts (~103 cells mL-1) and an abundance of Dethiobacter, a taxon within the phylum Clostridiales. Microcosm experiments were conducted with the purpose of monitoring carbon fixation, methanotrophy and metabolism of small organic compounds, such as acetate and formate, while tracing changes in fluid chemistry and microbial community composition. These experiments are expected to provide insight into the biogeochemical dynamics of the serpentinite subsurface at CROMO and represent a first step for developing metatranscriptomic and RNA-based Stable Isotope Probing (RNA-SIP) experiments to trace microbial activity at this site. [1] Brazelton et al. (2012) Frontiers in Microbiology 2:268
An in situ approach to study trace element partitioning in the laser heated diamond anvil cell
DOE Office of Scientific and Technical Information (OSTI.GOV)
Petitgirard, S.; Mezouar, M.; Borchert, M.
2012-01-15
Data on partitioning behavior of elements between different phases at in situ conditions are crucial for the understanding of element mobility especially for geochemical studies. Here, we present results of in situ partitioning of trace elements (Zr, Pd, and Ru) between silicate and iron melts, up to 50 GPa and 4200 K, using a modified laser heated diamond anvil cell (DAC). This new experimental set up allows simultaneous collection of x-ray fluorescence (XRF) and x-ray diffraction (XRD) data as a function of time using the high pressure beamline ID27 (ESRF, France). The technique enables the simultaneous detection of sample meltingmore » based to the appearance of diffuse scattering in the XRD pattern, characteristic of the structure factor of liquids, and measurements of elemental partitioning of the sample using XRF, before, during and after laser heating in the DAC. We were able to detect elements concentrations as low as a few ppm level (2-5 ppm) on standard solutions. In situ measurements are complimented by mapping of the chemical partitions of the trace elements after laser heating on the quenched samples to constrain the partitioning data. Our first results indicate a strong partitioning of Pd and Ru into the metallic phase, while Zr remains clearly incompatible with iron. This novel approach extends the pressure and temperature range of partitioning experiments derived from quenched samples from the large volume presses and could bring new insight to the early history of Earth.« less
Oyewumi, Oluyinka; Schreiber, Madeline E
2017-08-01
Trace elements are added to poultry feed to control infection and improve weight gain. However, the fate of these trace elements in poultry litter is poorly understood. Because poultry litter is applied as fertilizer in many agricultural regions, evaluation of the environmental processes that influence the mobility of litter-derived trace elements is critical for predicting if trace elements are retained in soil or released to water. This study examined the effect of dissolved organic carbon (DOC) in poultry litter leachate on the fate and transport of litter-derived elements (As, Cu, P and Zn) using laboratory column experiments with soil collected from the Delmarva Peninsula (Mid-Atlantic, USA), a region of intense poultry production. Results of the experiments showed that DOC enhanced the mobility of all of the studied elements. However, despite the increased mobility, 60-70% of Zn, As and P mass was retained within the soil. In contrast, almost all of the Cu was mobilized in the litter leachate experiments, with very little retention in soil. Overall, our results demonstrate that the mobility of As, Cu, Zn and P in soils which receive poultry litter application is strongly influenced by both litter leachate composition, specifically organic acids, and adsorption to soil. Results have implications for understanding fate and transport of trace elements released from litter application to soil water and groundwater, which can affect both human health and the environment. Copyright © 2017 Elsevier Ltd. All rights reserved.
U.S. biological experiments in space
NASA Technical Reports Server (NTRS)
Klein, H. P.
1981-01-01
The history of biologic experimentation in space is traced. Early balloon and rocket borne animals showed no abnormalities on the macroscale, and biosatellite launches with bacteria and amoebae revealed no microscopic dysfunctions. Adult Drosophila flies on board Cosmos spacecraft died with a shortened lifespan, while their offspring lived full lifespans. Green pepper plants grown in weightlessness showed a different orientation, but no physiological disturbances. Normal bone growth in rats has been found to almost cease after 11 days in space, and the mean life span of red blood cells decreases by four days. A series of experiments designed by U.S. scientists will be performed on primates provided and flown by the U.S.S.R. Finally, experiments on board Spacelab will involve determination of the persistence of circadian rhythms in bacteria and humans.
NASA Astrophysics Data System (ADS)
Dueker, M.; Clauson, K.; Yang, Q.; Umemoto, K.; Seltzer, A. M.; Zakharova, N. V.; Matter, J. M.; Stute, M.; Takahashi, T.; Goldberg, D.; O'Mullan, G. D.
2012-12-01
Despite growing appreciation for the importance of microbes in altering geochemical reactions in the subsurface, the microbial response to geological carbon sequestration injections and the role of microbes in altering metal mobilization following leakage scenarios in shallow aquifers remain poorly constrained. A Newark Basin test well was utilized in field experiments to investigate patterns of microbial succession following injection of CO2 saturated water into isolated aquifer intervals. Additionally, laboratory mesocosm experiments, including microbially-active and inactive (autoclave sterilized) treatments, were used to constrain the microbial role in mineral dissolution, trace metal release, and gas production (e.g. hydrogen and methane). Hydrogen production was detected in both sterilized and unsterilized laboratory mesocosm treatments, indicating abiotic hydrogen production may occur following CO2 leakage, and methane production was detected in unsterilized, microbially active mesocosms. In field experiments, a decrease in pH following injection of CO2 saturated aquifer water was accompanied by mobilization of trace elements (e.g. Fe and Mn), the production of hydrogen gas, and increased bacterial cell concentrations. 16S ribosomal RNA clone libraries, from samples collected before and after the test well injection, were compared in an attempt to link variability in geochemistry to changes in aquifer microbiology. Significant changes in microbial composition, compared to background conditions, were found following the test well injection, including a decrease in Proteobacteria, and an increased presence of Firmicutes, Verrucomicrobia, Acidobacteria and other microbes associated with iron reducing and syntrophic metabolism. The concurrence of increased microbial cell concentration, and rapid microbial community succession, with increased concentrations of hydrogen gas suggests that abiotically produced hydrogen may serve as an ecologically-relevant energy source stimulating changes in aquifer microbial communities immediately following CO2 leakage.
Jephcoat, Andrew P; Bouhifd, M Ali; Porcelli, Don
2008-11-28
The present state of the Earth evolved from energetic events that were determined early in the history of the Solar System. A key process in reconciling this state and the observable mantle composition with models of the original formation relies on understanding the planetary processing that has taken place over the past 4.5Ga. Planetary size plays a key role and ultimately determines the pressure and temperature conditions at which the materials of the early solar nebular segregated. We summarize recent developments with the laser-heated diamond anvil cell that have made possible extension of the conventional pressure limit for partitioning experiments as well as the study of volatile trace elements. In particular, we discuss liquid-liquid, metal-silicate (M-Sil) partitioning results for several elements in a synthetic chondritic mixture, spanning a wide range of atomic number-helium to iodine. We examine the role of the core as a possible host of both siderophile and trace elements and the implications that early segregation processes at deep magma ocean conditions have for current mantle signatures, both compositional and isotopic. The results provide some of the first experimental evidence that the core is the obvious replacement for the long-sought, deep mantle reservoir. If so, they also indicate the need to understand the detailed nature and scale of core-mantle exchange processes, from atomic to macroscopic, throughout the age of the Earth to the present day.
Pericytes of Multiple Organs Do Not Behave as Mesenchymal Stem Cells In Vivo.
Guimarães-Camboa, Nuno; Cattaneo, Paola; Sun, Yunfu; Moore-Morris, Thomas; Gu, Yusu; Dalton, Nancy D; Rockenstein, Edward; Masliah, Eliezer; Peterson, Kirk L; Stallcup, William B; Chen, Ju; Evans, Sylvia M
2017-03-02
Pericytes are widely believed to function as mesenchymal stem cells (MSCs), multipotent tissue-resident progenitors with great potential for regenerative medicine. Cultured pericytes isolated from distinct tissues can differentiate into multiple cell types in vitro or following transplantation in vivo. However, the cell fate plasticity of endogenous pericytes in vivo remains unclear. Here, we show that the transcription factor Tbx18 selectively marks pericytes and vascular smooth muscle cells in multiple organs of adult mouse. Fluorescence-activated cell sorting (FACS)-purified Tbx18-expressing cells behaved as MSCs in vitro. However, lineage-tracing experiments using an inducible Tbx18-CreERT2 line revealed that pericytes and vascular smooth muscle cells maintained their identity in aging and diverse pathological settings and did not significantly contribute to other cell lineages. These results challenge the current view of endogenous pericytes as multipotent tissue-resident progenitors and suggest that the plasticity observed in vitro or following transplantation in vivo arises from artificial cell manipulations ex vivo. Copyright © 2016 Elsevier Inc. All rights reserved.
Pancreatic β-Cells Express the Fetal Islet Hormone Gastrin in Rodent and Human Diabetes.
Dahan, Tehila; Ziv, Oren; Horwitz, Elad; Zemmour, Hai; Lavi, Judith; Swisa, Avital; Leibowitz, Gil; Ashcroft, Frances M; In't Veld, Peter; Glaser, Benjamin; Dor, Yuval
2017-02-01
β-Cell failure in type 2 diabetes (T2D) was recently proposed to involve dedifferentiation of β-cells and ectopic expression of other islet hormones, including somatostatin and glucagon. Here we show that gastrin, a stomach hormone typically expressed in the pancreas only during embryogenesis, is expressed in islets of diabetic rodents and humans with T2D. Although gastrin in mice is expressed in insulin + cells, gastrin expression in humans with T2D occurs in both insulin + and somatostatin + cells. Genetic lineage tracing in mice indicates that gastrin expression is turned on in a subset of differentiated β-cells after exposure to severe hyperglycemia. Gastrin expression in adult β-cells does not involve the endocrine progenitor cell regulator neurogenin3 but requires membrane depolarization, calcium influx, and calcineurin signaling. In vivo and in vitro experiments show that gastrin expression is rapidly eliminated upon exposure of β-cells to normal glucose levels. These results reveal the fetal hormone gastrin as a novel marker for reversible human β-cell reprogramming in diabetes. © 2017 by the American Diabetes Association.
Pancreatic β-Cells Express the Fetal Islet Hormone Gastrin in Rodent and Human Diabetes
Dahan, Tehila; Ziv, Oren; Horwitz, Elad; Zemmour, Hai; Lavi, Judith; Swisa, Avital; Leibowitz, Gil; Ashcroft, Frances M.; In’t Veld, Peter
2017-01-01
β-Cell failure in type 2 diabetes (T2D) was recently proposed to involve dedifferentiation of β-cells and ectopic expression of other islet hormones, including somatostatin and glucagon. Here we show that gastrin, a stomach hormone typically expressed in the pancreas only during embryogenesis, is expressed in islets of diabetic rodents and humans with T2D. Although gastrin in mice is expressed in insulin+ cells, gastrin expression in humans with T2D occurs in both insulin+ and somatostatin+ cells. Genetic lineage tracing in mice indicates that gastrin expression is turned on in a subset of differentiated β-cells after exposure to severe hyperglycemia. Gastrin expression in adult β-cells does not involve the endocrine progenitor cell regulator neurogenin3 but requires membrane depolarization, calcium influx, and calcineurin signaling. In vivo and in vitro experiments show that gastrin expression is rapidly eliminated upon exposure of β-cells to normal glucose levels. These results reveal the fetal hormone gastrin as a novel marker for reversible human β-cell reprogramming in diabetes. PMID:27864307
Strategic alliance, a way forward for violence against women: a case for the Special Cells, India.
Dave, Anjali
2013-10-01
This article attempts to describe the experiences of violated women and the struggles of social workers to contest violence against women in the Indian context. It begins with a brief account of an "indigenous model": the establishment of a service for violated women in India within the police force--the Special Cell on Violence Against Women. The article traces the strategic location, vision, growth, present position, expansion, and replication of the Special Cell in India, and discusses the necessity of working simultaneously with violated women, formal systems, and social structures; its contribution to the campaign for a Domestic Violence Act; and the resultant outcomes. The arduous nature of the work required for violated women and the women's own assessment of the Special Cells were accessed through a rigorous evaluation study, which is presented in the article, providing an answer and affirmation to the question: Why work with the Establishment--the State.
Mex3a marks slow-proliferating multilineage progenitors of the intestinal epithelium
Barriga, Francisco M.; Montagni, Elisa; Mana, Miyeko; Guillaumet-Adkins, Amy; Hernando-Momblona, Xavier; Sevillano, Marta; Rodriguez-Esteban, Gustavo; Mendez-Lago, Maria; Buczacki, Simon J. A.; Gut, Ivo; Gut, Marta; Winton, Douglas J.; Yilmaz, Omer; Stephan-Otto, Camille; Hein, Holger; Batlle, Eduard
2017-01-01
SUMMARY The intestinal epithelium is continuously regenerated by highly proliferative Lgr5+ intestinal stem cells (ISCs). The existence of a population of quiescent ISCs has been suggested yet its identity and features remain controversial. Here we describe that the expression of the RNA-binding protein Mex3a labels a subpopulation of Lgr5+ cells that divide less frequently and contribute to regenerate all intestinal lineages with slow kinetics. Single cell transcriptomic analysis revealed two classes of Lgr5-high cells, one of them defined by the Mex3a-expression program and by low levels of proliferation genes. Lineage tracing experiments show that large fraction of Mex3a+ cell population is continuously recalled into the rapidly dividing self-renewing ISC pool in homeostatic conditions. Chemotherapy and radiation target preferentially rapidly dividing Lgr5+ cells but spare the Mex3a-high/Lgr5+ population, which helps sustain the renewal of the intestinal epithelium during treatment. PMID:28285904
Interleaved Practice with Multiple Representations: Analyses with Knowledge Tracing Based Techniques
ERIC Educational Resources Information Center
Rau, Martina A.; Pardos, Zachary A.
2012-01-01
The goal of this paper is to use Knowledge Tracing to augment the results obtained from an experiment that investigated the effects of practice schedules using an intelligent tutoring system for fractions. Specifically, this experiment compared different practice schedules of multiple representations of fractions: representations were presented to…
Miwa, Hiroyuki; Era, Takumi
2018-01-29
Mesenchymal stem cells (MSCs) are somatic stem cells that can be derived from adult bone marrow (BM) and white adipose tissue (WAT), and that display multipotency and self-renewal capacity. Although MSCs are essential for tissue formation and have already been used in clinical therapy, the origins and markers of these cells remain unknown. In this study, we first investigated the developmental process of MSCs in mouse embryos using the gene encoding platelet-derived growth factor receptor α ( Pdgfra ) as a marker. We then traced cells expressing Pdgfra and other genes (brachyury, Sox1 and Pmx1 ) in various mutant mouse embryos until the adult stage. This tracing of MSC origins and destinies indicates that embryonic MSCs emerge in waves and that almost all adult BM MSCs and WAT MSCs originate from mesoderm and embryonic Pdgfrα-positive cells. Furthermore, we demonstrate that adult Pdgfrα-positive cells are involved in some pathological conditions. © 2018. Published by The Company of Biologists Ltd.
Baines, Stephen B.; Chen, Xi; Vogt, Stefan; Fisher, Nicholas S.; Twining, Benjamin S.; Landry, Michael R.
2016-01-01
Mesozooplankton production in high-nutrient low-chlorophyll regions of the ocean may be reduced if the trace element concentrations in their food are insufficient to meet growth and metabolic demands. We used elemental microanalysis (SXRF) of single-celled plankton to determine their trace metal contents during a series of semi-Lagrangian drift studies in an HNLC upwelling region, the Costa Rica Dome (CRD). Cells from the surface mixed layer had lower Fe:S but higher Zn:S and Ni:S than those from the subsurface chlorophyll maximum at 22–30 m. Diatom Fe:S values were typically 3-fold higher than those in flagellated cells. The ratios of Zn:C in flagellates and diatoms were generally similar to each other, and to co-occurring mesozooplankton. Estimated Fe:C ratios in flagellates were lower than those in co-occurring mesozooplankton, sometimes by more than 3-fold. In contrast, Fe:C in diatoms was typically similar to that in zooplankton. RNA:DNA ratios in the CRD were low compared with other regions, and were related to total autotrophic biomass and weakly to the discrepancy between Zn:C in flagellated cells and mesozooplankton tissues. Mesozooplankton may have been affected by the trace element content of their food, even though trace metal limitation of phytoplankton was modest at best. PMID:27275029
NASA Astrophysics Data System (ADS)
Kerekes, Ryan A.; Gleason, Shaun S.; Trivedi, Niraj; Solecki, David J.
2010-03-01
Segmentation, tracking, and tracing of neurons in video imagery are important steps in many neuronal migration studies and can be inaccurate and time-consuming when performed manually. In this paper, we present an automated method for tracing the leading and trailing processes of migrating neurons in time-lapse image stacks acquired with a confocal fluorescence microscope. In our approach, we first locate and track the soma of the cell of interest by smoothing each frame and tracking the local maxima through the sequence. We then trace the leading process in each frame by starting at the center of the soma and stepping repeatedly in the most likely direction of the leading process. This direction is found at each step by examining second derivatives of fluorescent intensity along curves of constant radius around the current point. Tracing terminates after a fixed number of steps or when fluorescent intensity drops below a fixed threshold. We evolve the resulting trace to form an improved trace that more closely follows the approximate centerline of the leading process. We apply a similar algorithm to the trailing process of the cell by starting the trace in the opposite direction. We demonstrate our algorithm on two time-lapse confocal video sequences of migrating cerebellar granule neurons (CGNs). We show that the automated traces closely approximate ground truth traces to within 1 or 2 pixels on average. Additionally, we compute line intensity profiles of fluorescence along the automated traces and quantitatively demonstrate their similarity to manually generated profiles in terms of fluorescence peak locations.
Viral vector-based tools advance knowledge of basal ganglia anatomy and physiology.
Sizemore, Rachel J; Seeger-Armbruster, Sonja; Hughes, Stephanie M; Parr-Brownlie, Louise C
2016-04-01
Viral vectors were originally developed to deliver genes into host cells for therapeutic potential. However, viral vector use in neuroscience research has increased because they enhance interpretation of the anatomy and physiology of brain circuits compared with conventional tract tracing or electrical stimulation techniques. Viral vectors enable neuronal or glial subpopulations to be labeled or stimulated, which can be spatially restricted to a single target nucleus or pathway. Here we review the use of viral vectors to examine the structure and function of motor and limbic basal ganglia (BG) networks in normal and pathological states. We outline the use of viral vectors, particularly lentivirus and adeno-associated virus, in circuit tracing, optogenetic stimulation, and designer drug stimulation experiments. Key studies that have used viral vectors to trace and image pathways and connectivity at gross or ultrastructural levels are reviewed. We explain how optogenetic stimulation and designer drugs used to modulate a distinct pathway and neuronal subpopulation have enhanced our mechanistic understanding of BG function in health and pathophysiology in disease. Finally, we outline how viral vector technology may be applied to neurological and psychiatric conditions to offer new treatments with enhanced outcomes for patients. Copyright © 2016 the American Physiological Society.
Viral vector-based tools advance knowledge of basal ganglia anatomy and physiology
Sizemore, Rachel J.; Seeger-Armbruster, Sonja; Hughes, Stephanie M.
2016-01-01
Viral vectors were originally developed to deliver genes into host cells for therapeutic potential. However, viral vector use in neuroscience research has increased because they enhance interpretation of the anatomy and physiology of brain circuits compared with conventional tract tracing or electrical stimulation techniques. Viral vectors enable neuronal or glial subpopulations to be labeled or stimulated, which can be spatially restricted to a single target nucleus or pathway. Here we review the use of viral vectors to examine the structure and function of motor and limbic basal ganglia (BG) networks in normal and pathological states. We outline the use of viral vectors, particularly lentivirus and adeno-associated virus, in circuit tracing, optogenetic stimulation, and designer drug stimulation experiments. Key studies that have used viral vectors to trace and image pathways and connectivity at gross or ultrastructural levels are reviewed. We explain how optogenetic stimulation and designer drugs used to modulate a distinct pathway and neuronal subpopulation have enhanced our mechanistic understanding of BG function in health and pathophysiology in disease. Finally, we outline how viral vector technology may be applied to neurological and psychiatric conditions to offer new treatments with enhanced outcomes for patients. PMID:26888111
PAX6 maintains β cell identity by repressing genes of alternative islet cell types.
Swisa, Avital; Avrahami, Dana; Eden, Noa; Zhang, Jia; Feleke, Eseye; Dahan, Tehila; Cohen-Tayar, Yamit; Stolovich-Rain, Miri; Kaestner, Klaus H; Glaser, Benjamin; Ashery-Padan, Ruth; Dor, Yuval
2017-01-03
Type 2 diabetes is thought to involve a compromised β cell differentiation state, but the mechanisms underlying this dysfunction remain unclear. Here, we report a key role for the TF PAX6 in the maintenance of adult β cell identity and function. PAX6 was downregulated in β cells of diabetic db/db mice and in WT mice treated with an insulin receptor antagonist, revealing metabolic control of expression. Deletion of Pax6 in β cells of adult mice led to lethal hyperglycemia and ketosis that were attributed to loss of β cell function and expansion of α cells. Lineage-tracing, transcriptome, and chromatin analyses showed that PAX6 is a direct activator of β cell genes, thus maintaining mature β cell function and identity. In parallel, we found that PAX6 binds promoters and enhancers to repress alternative islet cell genes including ghrelin, glucagon, and somatostatin. Chromatin analysis and shRNA-mediated gene suppression experiments indicated a similar function of PAX6 in human β cells. We conclude that reduced expression of PAX6 in metabolically stressed β cells may contribute to β cell failure and α cell dysfunction in diabetes.
PAX6 maintains β cell identity by repressing genes of alternative islet cell types
Swisa, Avital; Avrahami, Dana; Eden, Noa; Zhang, Jia; Feleke, Eseye; Dahan, Tehila; Cohen-Tayar, Yamit; Stolovich-Rain, Miri; Kaestner, Klaus H.; Glaser, Benjamin; Ashery-Padan, Ruth
2016-01-01
Type 2 diabetes is thought to involve a compromised β cell differentiation state, but the mechanisms underlying this dysfunction remain unclear. Here, we report a key role for the TF PAX6 in the maintenance of adult β cell identity and function. PAX6 was downregulated in β cells of diabetic db/db mice and in WT mice treated with an insulin receptor antagonist, revealing metabolic control of expression. Deletion of Pax6 in β cells of adult mice led to lethal hyperglycemia and ketosis that were attributed to loss of β cell function and expansion of α cells. Lineage-tracing, transcriptome, and chromatin analyses showed that PAX6 is a direct activator of β cell genes, thus maintaining mature β cell function and identity. In parallel, we found that PAX6 binds promoters and enhancers to repress alternative islet cell genes including ghrelin, glucagon, and somatostatin. Chromatin analysis and shRNA-mediated gene suppression experiments indicated a similar function of PAX6 in human β cells. We conclude that reduced expression of PAX6 in metabolically stressed β cells may contribute to β cell failure and α cell dysfunction in diabetes. PMID:27941241
Hunt, Pamela S; Barnet, Robert C
2015-09-01
Animal models of Fetal Alcohol Spectrum Disorders (FASD) afford the unique capacity to precisely control timing of alcohol exposure and alcohol exposure amounts in the developing animal. These models have powerfully informed neurophysiological alterations associated with fetal and perinatal alcohol. In two experiments presented here we expand use of the Pavlovian Trace Conditioning procedure to examine cognitive deficits and intervention strategies in a rat model of FASD. Rat pups were exposed to 5g/kg/day ethanol on postnatal days (PD) 4-9, simulating alcohol exposure in the third trimester in humans. During early adolescence, approximately PD 30, the rats were trained in the trace conditioning task in which a light conditioned stimulus (CS) and shock unconditioned stimulus (US) were paired but separated by a 10-s stimulus free trace interval. Learning was assessed in freezing behavior during shock-free tests. Experiment 1 revealed that neonatal ethanol exposure significantly impaired hippocampus-dependent trace conditioning relative to controls. In Experiment 2 a serial compound conditioning procedure known as 'gap filling' completely reversed the ethanol-induced deficit in trace conditioning. We also discuss prior data regarding the beneficial effects of supplemental choline and novel preliminary data regarding the pharmacological cognitive enhancer physostigmine, both of which mitigate the alcohol-induced cognitive deficit otherwise seen in trace conditioning controls. We suggest trace conditioning as a useful tool for characterizing some of the core cognitive deficits seen in FASD, and as a model for developing effective environmental as well as nutritional and pharmacological interventions. Copyright © 2014 Elsevier Inc. All rights reserved.
Hunt, Pamela S.; Barnet, Robert C.
2014-01-01
Animal models of Fetal Alcohol Spectrum Disorders (FASD) afford the unique capacity to precisely control timing of alcohol exposure and alcohol exposure amounts in the developing animal. These models have powerfully informed neurophysiological alterations associated with fetal and perinatal alcohol. In two experiments presented here we expand use of the Pavlovian Trace Conditioning procedure to examine cognitive deficits and intervention strategies in a rat model of FASD. Rat pups were exposed to 5 g/kg/day ethanol on postnatal days (PD) 4–9, simulating alcohol exposure in the third trimester in humans. During early adolescence, approximately PD 30, the rats were trained in the trace conditioning task in which a light conditioned stimulus (CS) and shock unconditioned stimulus (US) were paired but separated by a 10-s stimulus free trace interval. Learning was assessed in freezing behavior during shock-free tests. Experiment 1 revealed that neonatal ethanol exposure significantly impaired hippocampus-dependent trace conditioning relative to controls. In Experiment 2 a serial compound conditioning procedure known as ‘gap filling’ completely reversed the ethanol-induced deficit in trace conditioning. We also discuss prior data regarding the beneficial effects of supplemental choline and novel preliminary data regarding the pharmacological cognitive enhancer physostigmine, both of which mitigate the alcohol-induced cognitive deficit otherwise seen in trace conditioning controls. We suggest trace conditioning as a useful tool for characterizing some of the core cognitive deficits seen in FASD, and as a model for developing effective environmental as well as nutritional and pharmacological interventions. PMID:25477227
NASA Astrophysics Data System (ADS)
Gunina, Anna; Kuzyakov, Yakov; Glaser, Bruno
2015-04-01
Microorganisms control carbon (C) cycle and strongly contribute to formation of soil organic matter. Strong differences in the turnover of microbial groups and cellular compounds complicate the assessment of their contribution to microbial food webs and C sequestration in soil in situ. The uptake and incorporation of 13C labeled glucose by microbial groups were traced during 50 days after the labeling under field conditions. 13C was analysed: i) in the cytosolic pool by chloroform fumigation extraction, ii) in cell membranes by phospholipid fatty acids (PLFA), iii) in cell walls by amino sugars, and iv) remaining in bulk soil. This allowed tracing C in microbial groups as well as cellular compounds. Mean residence times (MRT) of C in PLFA and the cytosol were 47 and 150 days, respectively. Such long cytosol MRT depends on its heterogeneous composition, which includes high and low molecular weight organics. Amino sugars were mainly originated from microbial residues and thus, observation periods higher than 1 year are required for estimation of their MRT. Relative 13C incorporation (13C portion in total pool C) was the highest for PLFAs (~1.5% at day 3), whereas 13C content of the cytosol and amino sugars was one and two orders of magnitude less, respectively. Relative 13C incorporation into amino sugars of living microorganisms showed only 0.57% on day 3. Therefore, the turnover of cell membrane components is two times faster than that of cell walls, even in living microorganisms. Both PLFAs and amino sugars showed that glucose C was preferentially used by bacteria. 13C incorporation into bacterial cell walls and membranes decreased with time, but increased or remained constant for fungi, reflecting faster turnover of bacteria than fungi. Consequently, bacteria contribute more to the decomposition of low molecular weight organics, whereas fungi consume bacterial products or necromass and contribute more to long-term C stabilisation. Thus, tracing of 13C in cellular compounds with contrasting turnover provides key information to C fluxes through the soil microbial food-web and elucidates the role of distinct groups as well as individual cellular compartments in SOM formation and C sequestration.
An engram found? Evaluating the evidence from fruit flies.
Gerber, Bertram; Tanimoto, Hiromu; Heisenberg, Martin
2004-12-01
Is it possible to localize a memory trace to a subset of cells in the brain? If so, it should be possible to show: first, that neuronal plasticity occurs in these cells. Second, that neuronal plasticity in these cells is sufficient for memory. Third, that neuronal plasticity in these cells is necessary for memory. Fourth, that memory is abolished if these cells cannot provide output during testing. And fifth, that memory is abolished if these cells cannot receive input during training. With regard to olfactory learning in flies, we argue that the notion of the olfactory memory trace being localized to the Kenyon cells of the mushroom bodies is a reasonable working hypothesis.
Radyk, Megan D; Burclaff, Joseph; Willet, Spencer G; Mills, Jason C
2018-03-01
Spasmolytic polypeptide-expressing metaplasia (SPEM) develops in patients with chronic atrophic gastritis due to infection with Helicobacter pylori; it might be a precursor to intestinal metaplasia and gastric adenocarcinoma. Lineage tracing experiments of the gastric corpus in mice have not established whether SPEM derives from proliferating stem cells or differentiated, post-mitotic zymogenic chief cells in the gland base. We investigated whether differentiated cells can give rise to SPEM using a nongenetic approach in mice. Mice were given intraperitoneal injections of 5-fluorouracil, which blocked gastric cell proliferation, plus tamoxifen to induce SPEM. Based on analyses of molecular and histologic markers, we found SPEM developed even in the absence of cell proliferation. SPEM therefore did not arise from stem cells. In histologic analyses of gastric resection specimens from 10 patients with adenocarcinoma, we found normal zymogenic chief cells that were transitioning into SPEM cells only in gland bases, rather than the proliferative stem cell zone. Our findings indicate that SPEM can arise by direct reprogramming of existing cells-mainly of chief cells. Copyright © 2018 AGA Institute. Published by Elsevier Inc. All rights reserved.
Dielectric constant of atomic fluids with variable polarizability
Alder, B. J.; Beers, J. C.; Strauss, H. L.; Weis, J. J.
1980-01-01
The Clausius-Mossotti function for the dielectric constant is expanded in terms of single atom and pair polarizabilities, leading to contributions that depend on both the trace and the anisotropy of the pair-polarizability tensor. The short-range contribution of the anisotropic part to the pair polarizabilities has previously been obtained empirically from light scattering experiments, whereas the trace contribution is now empirically determined by comparison to dielectric experiments. For helium, the short-range trace part agrees well with electronic structure calculations, whereas for argon qualitative agreement is achieved. PMID:16592830
Dielectric constant of atomic fluids with variable polarizability.
Alder, B J; Beers, J C; Strauss, H L; Weis, J J
1980-06-01
The Clausius-Mossotti function for the dielectric constant is expanded in terms of single atom and pair polarizabilities, leading to contributions that depend on both the trace and the anisotropy of the pair-polarizability tensor. The short-range contribution of the anisotropic part to the pair polarizabilities has previously been obtained empirically from light scattering experiments, whereas the trace contribution is now empirically determined by comparison to dielectric experiments. For helium, the short-range trace part agrees well with electronic structure calculations, whereas for argon qualitative agreement is achieved.
Microbial Influences on Trace Metal Cycling in a Meromictic Lake, Fayetteville Green Lake, NY
NASA Astrophysics Data System (ADS)
Zerkle, A. L.; House, C.; Kump, L.
2002-12-01
Microorganisms can exist in aquatic environments at very high cell densities of up to 1011 cells/L, and can accumulate significant quantities of trace metals. Bacteria actively take up bioactive trace metals, including Fe, Zn, Mn, Co, Ni, Cu, and Mo, which function as catalytic centers in metalloproteins and metal-activated enzymes involved in virtually all cellular functions. In addition, bacteria may catalyze the release of trace metals from inorganic substrates by processes such as the reduction of iron and manganese oxides, suggesting that trace metal distributions within a natural environment dominated by microbial processes may be controlled primarily by microbial ecology. Fayetteville Green Lake (FGL), NY, is a permanently stratified meromictic lake that has a well-oxygenated surface water mass (mixolimnion) overlying a relatively stagnant, anoxic deep water mass (monimolimnion). A chemocline separates the water masses at around 20m depth, where oxygen concentrations decrease and sulfate and methane concentrations increase. In addition, previous studies have indicated that trace metals such as V, Cr, Co, Mn, and Fe reach elevated concentrations at the chemocline. Using fluorescent in situ hybridization (FISH) of FGL samples from depths of up to 40m with bacterial and archaeal probes, we have shown that fluctuating redox conditions within the FGL water column correlate with significant variations in the composition and distribution of microbial populations with depth. The mixolimnion is dominated by Eubacteria, with increasing concentrations of Archaea in the lower anoxic zone. Increases in microbial cell densities coincide with increases in trace metals at the chemocline, suggesting microbial activity may be responsible for trace metal release at this boundary. 16S rRNA PCR cloning techniques are currently being used to identify dominant microbial populations at various levels within the FGL water column. Future studies will focus on the potential for these dominant microorganisms to influence trace metal cycling and bioavailability in the FGL water column.
Yin, Hang; Pasut, Alessandra; Soleimani, Vahab D.; Bentzinger, C. Florian; Antoun, Ghadi; Thorn, Stephanie; Seale, Patrick; Fernando, Pasan; van IJcken, Wilfred; Grosveld, Frank; Dekemp, Robert A.; Boushel, Robert; Harper, Mary-Ellen; Rudnicki, Michael A.
2013-01-01
SUMMARY Brown adipose tissue (BAT) is an energy-dispensing thermogenic tissue that plays an important role in balancing energy metabolism. Lineage-tracing experiments indicate that brown adipocytes are derived from myogenic progenitors during embryonic development. However, adult skeletal muscle stem cells (satellite cells) have long been considered uniformly determined toward the myogenic lineage. Here, we report that adult satellite cells give rise to brown adipocytes and that microRNA-133 regulates the choice between myogenic and brown adipose determination by targeting the 3′UTR of Prdm16. Antagonism of microRNA-133 during muscle regeneration increases uncoupled respiration, glucose uptake, and thermogenesis in local treated muscle and augments whole-body energy expenditure, improves glucose tolerance, and impedes the development of diet-induced obesity. Finally, we demonstrate that miR-133 levels are downregulated in mice exposed to cold, resulting in de novo generation of satellite cell-derived brown adipocytes. Therefore, microRNA-133 represents an important therapeutic target for the treatment of obesity. PMID:23395168
Analytical Methods for Trace Metals. Training Manual.
ERIC Educational Resources Information Center
Office of Water Program Operations (EPA), Cincinnati, OH. National Training and Operational Technology Center.
This training manual presents material on the theoretical concepts involved in the methods listed in the Federal Register as approved for determination of trace metals. Emphasis is on laboratory operations. This course is intended for chemists and technicians with little or no experience in analytical methods for trace metals. Students should have…
A dual-trace model for visual sensory memory.
Cappiello, Marcus; Zhang, Weiwei
2016-11-01
Visual sensory memory refers to a transient memory lingering briefly after the stimulus offset. Although previous literature suggests that visual sensory memory is supported by a fine-grained trace for continuous representation and a coarse-grained trace of categorical information, simultaneous separation and assessment of these traces can be difficult without a quantitative model. The present study used a continuous estimation procedure to test a novel mathematical model of the dual-trace hypothesis of visual sensory memory according to which visual sensory memory could be modeled as a mixture of 2 von Mises (2VM) distributions differing in standard deviation. When visual sensory memory and working memory (WM) for colors were distinguished using different experimental manipulations in the first 3 experiments, the 2VM model outperformed Zhang and Luck (2008) standard mixture model (SM) representing a mixture of a single memory trace and random guesses, even though SM outperformed 2VM for WM. Experiment 4 generalized 2VM's advantages of fitting visual sensory memory data over SM from color to orientation. Furthermore, a single trace model and 4 other alternative models were ruled out, suggesting the necessity and sufficiency of dual traces for visual sensory memory. Together these results support the dual-trace model of visual sensory memory and provide a preliminary inquiry into the nature of information loss from visual sensory memory to WM. (PsycINFO Database Record (c) 2016 APA, all rights reserved).
Genetic Lineage Tracing of Non-Myocyte Population by Dual Recombinases.
Li, Yan; He, Lingjuan; Huang, Xiuzhen; Issa Bhaloo, Shirin; Zhao, Huan; Zhang, Shaohua; Pu, Wenjuan; Tian, Xueying; Li, Yi; Liu, Qiaozhen; Yu, Wei; Zhang, Libo; Liu, Xiuxiu; Liu, Kuo; Tang, Juan; Zhang, Hui; Cai, Dongqing; Adams, Ralf H; Xu, Qingbo; Lui, Kathy O; Zhou, Bin
2018-04-26
Background -Whether the adult mammalian heart harbors cardiac stem cells (CSCs) for regeneration of cardiomyocytes is an important yet contentious topic in the field of cardiovascular regeneration. The putative myocyte stem cell populations recognized without specific cell markers such as the cardiosphere-derived cells or with markers such as Sca1 + , Bmi1 + , Isl1 + or Abcg2 + CSCs have been reported. Moreover, it remains unclear whether putative CSCs with unknown or unidentified markers exist and give rise to de novo cardiomyocytes in the adult heart. Methods -To address this question without relying on a particular stem cell marker, we developed a new genetic lineage tracing system to label all non-myocyte populations that contain putative CSCs. Using dual lineage tracing system, we assessed if non-myocytes generated any new myocytes during embryonic development, adult homeostasis and after myocardial infarction. Skeletal muscle was also examined after injury for internal control of new myocytes generation from non-myocytes. Results -By this stem cell marker-free and dual recombinases-mediated cell tracking approach, our fate mapping data show that new myocytes arise from non-myocytes in the embryonic heart, but not in the adult heart during homeostasis or after myocardial infarction. As positive control, our lineage tracing system detected new myocytes derived from non-myocytes in the skeletal muscle after injury. Conclusions -This study provides in vivo genetic evidence for non-myocyte to myocyte conversion in embryonic but not adult heart, arguing again the myogenic potential of putative stem cell populations for cardiac regeneration in the adult stage. This study also provides a new genetic strategy to identify endogenous stem cells, if any, in other organ systems for tissue repair and regeneration.
Kalmbach, Brian; Chitwood, Raymond A.; Mauk, Michael D.
2012-01-01
We have addressed the source and nature of the persistent neural activity that bridges the stimulus-free gap between the conditioned stimulus (CS) and unconditioned stimulus (US) during trace eyelid conditioning. Previous work has demonstrated that this persistent activity is necessary for trace eyelid conditioning: CS-elicited activity in mossy fiber inputs to the cerebellum does not extend into the stimulus-free trace interval, which precludes the cerebellar learning that mediates conditioned response expression. In behaving rabbits we used in vivo recordings from a region of medial prefrontal cortex (mPFC) that is necessary for trace eyelid conditioning to test the hypothesis that neurons there generate activity that persists beyond CS offset. These recordings revealed two patterns of activity during the trace interval that would enable cerebellar learning. Activity in some cells began during the tone CS and persisted to overlap with the US, whereas in other cells, activity began during the stimulus-free trace interval. Injection of anterograde tracers into this same region of mPFC revealed dense labeling in the pontine nuclei, where recordings also revealed tone-evoked persistent activity during trace conditioning. These data suggest a corticopontine pathway that provides an input to the cerebellum during trace conditioning trials that bridges the temporal gap between the CS and US to engage cerebellar learning. As such, trace eyelid conditioning represents a well-characterized and experimentally tractable system that can facilitate mechanistic analyses of cortical persistent activity and how it is used by downstream brain structures to influence behavior. PMID:21957220
Zhou, Sihui; Zhang, Caiying; Xiao, Qingyang; Zhuang, Yu; Gu, Xiaolong; Yang, Fan; Xing, Chenghong; Hu, Guoliang; Cao, Huabin
2016-11-01
Molybdenum (Mo) is an essential trace element for animals and human beings. However, the negative effects on rumen function and distribution of trace elements in tissues induced by excessive Mo have not been well understood. Therefore, the purpose of present study was to investigate the impact of Mo on rumen microbiota, distribution of trace elements in various organs, and hematological parameters of goats. A total of 36 goats were randomly distributed into three groups with equal number and low-Mo and high-Mo groups were orally administered ammonium molybdate at 15 and 45 mg · Mo · kg -1 · BW respectively, while the control group received corresponding quantitative deionized water. The results showed that the total number of ciliate and protozoa protein concentration decreased significantly (P < 0.01) on days 25 and 50. Concentrations of ammonia nitrogen and bacterial protein were significantly higher (P < 0.05) in low-Mo group, while they were lower (P < 0.05) in high-Mo group than the control group on days 25 and 50. In addition, Mo accumulated in serum and all detected tissues. Copper (Cu) and zinc (Zn) contents significantly decreased (P < 0.05) in hair and serum on days 25 and 50, while Cu contents increased (P < 0.05) and the change of Zn contents were not obvious (P > 0.05) in other tissues on days 25 and 50. Besides, there was no obvious variation in iron (Fe) contents during whole experiment period (P > 0.05). Furthermore, excessive Mo content had no significant effect on red blood cell (RBC) counts and hemoglobin (HGB) concentration (P > 0.05) on days 25 and 50, while white blood cell (WBC) counts increased significantly (P < 0.05) on day 50. These results indicated that excessive Mo content could impact the balance of ruminal microorganisms and interfere with the absorption and distribution of Mo and Cu mainly.
Cai, Yafan; Wang, Jungang; Zhao, Yubin; Zhao, Xiaoling; Zheng, Zehui; Wen, Boting; Cui, Zongjun; Wang, Xiaofen
2018-09-01
Trace elements were commonly used as additives to facilitate anaerobic digestion. However, their addition is often blind because of the complexity of reaction conditions, which has impeded their widespread application. Therefore, this study was conducted to evaluate deficiencies in trace elements during anaerobic digestion by establishing relationships between changes in trace element bioavailability (the degree to which elements are available for interaction with biological systems) and digestion performance. To accomplish this, two batch experiments were conducted. In the first, sequential extraction was used to detect changes in trace element fractions and then to evaluate trace element bioavailability in the whole digestion cycle. In the second batch experiment, trace elements (Co, Fe, Cu, Zn, Mn, Mo and Se) were added to the reaction system at three concentrations (low, medium and high) and their effects were monitored. The results showed that sequential extraction was a suitable method for assessment of the bioavailability of trace elements (appropriate coefficient of variation and recovery rate). The results revealed that Se had the highest (44.2%-70.9%) bioavailability, while Fe had the lowest (1.7%-3.0%). A lack of trace elements was not directly related to their absolute bioavailability, but was instead associated with changes in their bioavailability throughout the digestion cycle. Trace elements were insufficient when their bioavailability was steady or increased over the digestion cycle. These results indicate that changes in trace element bioavailability during the digestion cycle can be used to predict their deficiency. Copyright © 2018 Elsevier Ltd. All rights reserved.
Rolland, Anne-Sophie; Karachi, Carine; Muriel, Marie-Paule; Hirsch, Etienne C; François, Chantal
2011-08-01
The locomotor area has recently emerged as a target for deep brain stimulation to lessen gait disturbances in advanced parkinsonian patients. An important step in choosing this target is to define anatomical limits of its 2 components, the pedunculopontine nucleus and the cuneiform nucleus, their connections with the basal ganglia, and their output descending pathway. Based on the hypothesis that pedunculopontine nucleus controls locomotion whereas cuneiform nucleus controls axial posture, we analyzed whether both nuclei receive inputs from the internal pallidum and substantia nigra using anterograde and retrograde tract tracing in monkeys. We also examined whether these nuclei convey descending projections to the reticulospinal pathway. Pallidal terminals were densely distributed and restricted to the pedunculopontine nucleus, whereas nigral terminals were diffusely observed in the whole extent of both the pedunculopontine nucleus and the cuneiform nucleus. Moreover, nigral terminals formed symmetric synapses with pedunculopontine nucleus and cuneiform nucleus dendrites. Retrograde tracing experiments confirmed these results because labeled cell bodies were observed in both the internal pallidum and substantia nigra after pedunculopontine nucleus injection, but only in the substantia nigra after cuneiform nucleus injection. Furthermore, anterograde tracing experiments revealed that the pedunculopontine nucleus and cuneiform nucleus project to large portions of the pontomedullary reticular formation. This is the first anatomical evidence that the internal pallidum and the substantia nigra control different parts of the brain stem and can modulate the descending reticulospinal pathway in primates. These findings support the functional hypothesis that the nigro-cuneiform nucleus pathway could control axial posture whereas the pallido-pedunculopontine nucleus pathway could modulate locomotion. Copyright © 2011 Movement Disorder Society.
Design and research of built-in sample cell with multiple optical reflections
NASA Astrophysics Data System (ADS)
Liu, Jianhui; Wang, Shuyao; Lv, Jinwei; Liu, Shuyang; Zhou, Tao; Jia, Xiaodong
2017-10-01
In the field of trace gas measurement, with the characteristics of high sensitivity, high selectivity and rapid detection, tunable diode laser absorption spectroscopy (TDLAS) is widely used in industrial process and trace gas pollution monitoring. Herriott cell is a common form of multiple reflections of the sample cell, the structure of the Herriott cell is relatively simple, which be used to application of trace gas absorption spectroscopy. In the pragmatic situation, the gas components are complicated, and the continuous testing process for a long time can lead to different degree of pollution and corrosion for the reflector in the sample cell. If the mirror is not cleaned up in time, it will have a great influence on the detection accuracy. In order to solve this problem in the process of harsh environment detection, this paper presents a design of the built-in sample cell to avoid the contact of gas and the mirror, thereby effectively reducing corrosion pollution. If there is optical pollution, direct replacement of the built-in optical sample cell can easily to be disassembled, and cleaned. The advantage of this design is long optical path, high precision, cost savings and so on.
Ogawa, Ryo; Ma, Yuran; Yamaguchi, Masahiko; Ito, Takahito; Watanabe, Yoko; Ohtani, Takuji; Murakami, Satoshi; Uchida, Shizuka; De Gaspari, Piera; Uezumi, Akiyoshi; Nakamura, Miki; Miyagoe-Suzuki, Yuko; Tsujikawa, Kazutake; Hashimoto, Naohiro; Braun, Thomas; Tanaka, Teruyuki; Takeda, Shin'ichi; Yamamoto, Hiroshi; Fukada, So-Ichiro
2015-01-01
Muscle satellite cells are indispensable for muscle regeneration, but the functional diversity of their daughter cells is unknown. Here, we show that many Pax7(+)MyoD(-) cells locate both beneath and outside the basal lamina during myofiber maturation. A large majority of these Pax7(+)MyoD(-) cells are not self-renewed satellite cells, but have different potentials for both proliferation and differentiation from Pax7(+)MyoD(+) myoblasts (classical daughter cells), and are specifically marked by expression of the doublecortin (Dcx) gene. Transplantation and lineage-tracing experiments demonstrated that Dcx-expressing cells originate from quiescent satellite cells and that the microenvironment induces Dcx in myoblasts. Expression of Dcx seems to be necessary for myofiber maturation because Dcx-deficient mice exhibited impaired myofiber maturation resulting from a decrease in the number of myonuclei. Furthermore, in vitro and in vivo studies suggest that one function of Dcx in myogenic cells is acceleration of cell motility. These results indicate that Dcx is a new marker for the Pax7(+)MyoD(-) subpopulation, which contributes to myofiber maturation during muscle regeneration. © 2015. Published by The Company of Biologists Ltd.
Magistretti, Jacopo; Ragsdale, David S; Alonso, Angel
1999-01-01
Single Na+ channel activity was recorded in patch-clamp, cell-attached experiments performed on dendritic processes of acutely isolated principal neurones from rat entorhinal-cortex layer II. The distances of the recording sites from the soma ranged from ≈20 to ≈100 μm.Step depolarisations from holding potentials of −120 to −100 mV to test potentials of −60 to +10 mV elicited Na+ channel openings in all of the recorded patches (n= 16).In 10 patches, besides transient Na+ channel openings clustered within the first few milliseconds of the depolarising pulses, prolonged and/or late Na+ channel openings were also regularly observed. This ‘persistent’ Na+ channel activity produced net inward, persistent currents in ensemble-average traces, and remained stable over the entire duration of the experiments (≈9 to 30 min).Two of these patches contained <= 3 channels. In these cases, persistent Na+ channel openings could be attributed to the activity of one single channel.The voltage dependence of persistent-current amplitude in ensemble-average traces closely resembled that of whole-cell, persistent Na+ current expressed by the same neurones, and displayed the same characteristic low threshold of activation.Dendritic, persistent Na+ channel openings had relatively high single-channel conductance (≈20 pS), similar to what is observed for somatic, persistent Na+ channels.We conclude that a stable, persistent Na+ channel activity is expressed by proximal dendrites of entorhinal-cortex layer II principal neurones, and can contribute a significant low-threshold, persistent Na+ current to the dendritic processing of excitatory synaptic inputs. PMID:10601494
NASA Astrophysics Data System (ADS)
Chan, E. W.; Kessler, J. D.; Redmond, M. C.; Shiller, A. M.; Arrington, E. C.; Valentine, D. L.; Colombo, F.
2015-12-01
Many studies of microbially mediated aerobic methane oxidation in oceanic environments have examined the many different factors that control the rates of oxidation. However, there is debate on how quickly methane is oxidized once a microbial population is established and what factor(s) are limiting in these types of environments. These factors include the availability of CH4, O2, trace metals, nutrients, and the density of cell population. Limits to these factors can also control the temporal aspects of a methane oxidation event. In order to look at this process in its entirety and with higher temporal resolution, a mesocosm incubation system was developed with a Dissolved Gas Analyzer System (DGAS) coupled with a set of analytical tools to monitor aerobic methane oxidation in real time. With the addition of newer laser spectroscopy techniques (cavity ringdown spectroscopy), stable isotope fractionation caused by microbial processes can also be examined on a real time and automated basis. Cell counting, trace metal, nutrient, and DNA community analyses have also been carried out in conjunction with these mesocosm samples to provide a clear understanding of the biology in methane oxidation dynamics. This poster will detail the techniques involved to provide insights into the chemical and isotopic kinetics controlling aerobic methane oxidation. Proof of concept applications will be presented from seep sites in the Hudson Canyon and the Sleeping Dragon seep field, Mississippi Canyon 118 (MC 118). This system was used to conduct mesocosm experiments to examine methane consumption, O2 consumption, nutrient consumption, and biomass production.
Kast, Ryan J; Wu, Hsiao-Huei; Williams, Piper; Gaspar, Patricia; Levitt, Pat
2017-05-17
Molecular characterization of neurons across brain regions has revealed new taxonomies for understanding functional diversity even among classically defined neuronal populations. Neuronal diversity has become evident within the brain serotonin (5-HT) system, which is far more complex than previously appreciated. However, until now it has been difficult to define subpopulations of 5-HT neurons based on molecular phenotypes. We demonstrate that the MET receptor tyrosine kinase (MET) is specifically expressed in a subset of 5-HT neurons within the caudal part of the dorsal raphe nuclei (DRC) that is encompassed by the classic B6 serotonin cell group. Mapping from embryonic day 16 through adulthood reveals that MET is expressed almost exclusively in the DRC as a condensed, paired nucleus, with an additional sparse set of MET+ neurons scattered within the median raphe. Retrograde tracing experiments reveal that MET-expressing 5-HT neurons provide substantial serotonergic input to the ventricular/subventricular region that contains forebrain stem cells, but do not innervate the dorsal hippocampus or entorhinal cortex. Conditional anterograde tracing experiments show that 5-HT neurons in the DRC/B6 target additional forebrain structures such as the medial and lateral septum and the ventral hippocampus. Molecular neuroanatomical analysis identifies 14 genes that are enriched in DRC neurons, including 4 neurotransmitter/neuropeptide receptors and 2 potassium channels. These analyses will lead to future studies determining the specific roles that 5-HT MET+ neurons contribute to the broader set of functions regulated by the serotonergic system.
Schätzlein, Martina Palomino; Becker, Johanna; Schulze-Sünninghausen, David; Pineda-Lucena, Antonio; Herance, José Raul; Luy, Burkhard
2018-04-01
Isotope labeling enables the use of 13 C-based metabolomics techniques with strongly improved resolution for a better identification of relevant metabolites and tracing of metabolic fluxes in cell and animal models, as required in fluxomics studies. However, even at high NMR-active isotope abundance, the acquisition of one-dimensional 13 C and classical two-dimensional 1 H, 13 C-HSQC experiments remains time consuming. With the aim to provide a shorter, more efficient alternative, herein we explored the ALSOFAST-HSQC experiment with its rapid acquisition scheme for the analysis of 13 C-labeled metabolites in complex biological mixtures. As an initial step, the parameters of the pulse sequence were optimized to take into account the specific characteristics of the complex samples. We then applied the fast two-dimensional experiment to study the effect of different kinds of antioxidant gold nanoparticles on a HeLa cancer cell model grown on 13 C glucose-enriched medium. As a result, 1 H, 13 C-2D correlations could be obtained in a couple of seconds to few minutes, allowing a simple and reliable identification of various 13 C-enriched metabolites and the determination of specific variations between the different sample groups. Thus, it was possible to monitor glucose metabolism in the cell model and study the antioxidant effect of the coated gold nanoparticles in detail. Finally, with an experiment time of only half an hour, highly resolved 1 H, 13 C-HSQC spectra using the ALSOFAST-HSQC pulse sequence were acquired, revealing the isotope-position-patterns of the corresponding 13 C-nuclei from carbon multiplets. Graphical abstract Fast NMR applied to metabolomics and fluxomics studies with gold nanoparticles.
Laser Ray Tracing in a Parallel Arbitrary Lagrangian-Eulerian Adaptive Mesh Refinement Hydrocode
DOE Office of Scientific and Technical Information (OSTI.GOV)
Masters, N D; Kaiser, T B; Anderson, R W
2009-09-28
ALE-AMR is a new hydrocode that we are developing as a predictive modeling tool for debris and shrapnel formation in high-energy laser experiments. In this paper we present our approach to implementing laser ray-tracing in ALE-AMR. We present the equations of laser ray tracing, our approach to efficient traversal of the adaptive mesh hierarchy in which we propagate computational rays through a virtual composite mesh consisting of the finest resolution representation of the modeled space, and anticipate simulations that will be compared to experiments for code validation.
Effects of Humidity Swings on Adsorption Columns for Air Revitalization: Modeling and Experiments
NASA Technical Reports Server (NTRS)
LeVan, M. Douglas; Finn, John E.
1997-01-01
Air purification systems are necessary to provide clean air in the closed environments aboard spacecraft. Trace contaminants are removed using adsorption. One major factor concerning the removal of trace contaminants is relative humidity. Water can reduce adsorption capacity and, due to constant fluctuations, its presence is difficult to incorporate into adsorption column designs. The purpose of the research was to allow for better design techniques in trace contaminant adsorption systems, especially for feeds with water present. Experiments and mathematical modeling research on effects of humidity swings on adsorption columns for air revitalization were carried out.
Sawano, Makoto; Shimouchi, Akito
2010-09-01
Past studies have suggested that some carbon monoxide (CO) moves from blood haemoglobin to tissue cells and that mitochondrial cytochrome c oxidase oxidizes CO to carbon dioxide (CO(2)). However, no study has demonstrated this redistribution and oxidization of CO under physiological conditions. The objective of this study was to trace the redistribution and oxidization of CO in the human body by detecting (13)CO(2) production after the inhalation of (13)CO. In Experiment 1, we asked a healthy subject to inhale 50 ppm (13)CO gas. In Experiment 2, we circulated heparinized human blood in a cardio-pulmonary bypass circuit and supplied 50 ppm (13)CO gas to the oxygenator. We sequentially sampled exhaled and output gases and measured the (13)CO(2)/(12)CO(2) ratios. In Experiment 1, the exhaled (13)CO(2)/(12)CO(2) ratio increased significantly between 4 to 31 h of (13)CO inhalation. In Experiment 2, the output (13)CO(2)/(12)CO(2) ratio showed no significant increase within 36 h of (13)CO input. Experiment 1 demonstrated the oxidization of CO in the human body under physiological conditions. Experiment 2 confirmed that oxidization does not occur in the circulating blood and indicated the redistribution of CO from blood carboxyhaemoglobin to tissue cells.
Ray tracing a three dimensional scene using a grid
Wald, Ingo; Ize, Santiago; Parker, Steven G; Knoll, Aaron
2013-02-26
Ray tracing a three-dimensional scene using a grid. One example embodiment is a method for ray tracing a three-dimensional scene using a grid. In this example method, the three-dimensional scene is made up of objects that are spatially partitioned into a plurality of cells that make up the grid. The method includes a first act of computing a bounding frustum of a packet of rays, and a second act of traversing the grid slice by slice along a major traversal axis. Each slice traversal includes a first act of determining one or more cells in the slice that are overlapped by the frustum and a second act of testing the rays in the packet for intersection with any objects at least partially bounded by the one or more cells overlapped by the frustum.
Dissolution of Platinum in the Operational Range of Fuel Cells
Keeley, Gareth P.; Geiger, Simon; Zeradjanin, Aleksandar R.; Hodnik, Nejc; Kulyk, Nadiia
2015-01-01
Abstract One of the most important practical issues in low‐temperature fuel‐cell catalyst degradation is platinum dissolution. According to the literature, it initiates at 0.6–0.9 VRHE, whereas previous time‐ and potential‐resolved inductively coupled plasma mass spectrometry (ICP–MS) experiments, however, revealed dissolution onset at only 1.05 VRHE. In this manuscript, the apparent discrepancy is addressed by investigating bulk and nanoparticulated catalysts. It is shown that, given enough time for accumulation, traces of platinum can be detected at potentials as low as 0.85 VRHE. At these low potentials, anodic dissolution is the dominant process, whereas, at more positive potentials, more platinum dissolves during the oxide reduction after accumulation. Interestingly, the potential and time dissolution dependence is similar for both types of electrode. Dissolution processes are discussed with relevance to fuel‐cell operation and plausible dissolution mechanisms are considered. PMID:27525206
The Road Less Travelled: Tracing the Path of First-Generation Students from Rural Areas to College
ERIC Educational Resources Information Center
Hodsdon, Michelle Caron
2012-01-01
Narrative inquiry was used to trace the educational journeys of 11 first-generation university students from rural areas of Colorado in an effort to identify the experiences, beliefs, and people that impacted their decision to attend a 4-year institution. Students were asked to convey their experiences growing up within the contexts of their…
Lin, Yi-Zhen; Ou, Da-Liang; Chang, Hsin-Yuan; Lin, Wei-Yu; Hsu, Chiun; Chang, Po-Ling
2017-09-01
The family of microRNAs (miRNAs) not only plays an important role in gene regulation but is also useful for the diagnosis of diseases. A reliable method with high sensitivity may allow researchers to detect slight fluctuations in ultra-trace amounts of miRNA. In this study, we propose a sensitive imaging method for the direct probing of miR-10b (miR-10b-3p, also called miR-10b*) and its target ( HOXD10 mRNA) in fixed cells based on the specific recognition of molecular beacons combined with highly inclined and laminated optical sheet (HILO) fluorescence microscopy. The designed dye-quencher-labelled molecular beacons offer excellent efficiencies of fluorescence resonance energy transfer that allow us to detect miRNA and the target mRNA simultaneously in hepatocellular carcinoma cells using HILO fluorescence microscopy. Not only can the basal trace amount of miRNA be observed in each individual cell, but the obtained images also indicate that this method is useful for monitoring the fluctuations in ultra-trace amounts of miRNA when the cells are transfected with a miRNA precursor or a miRNA inhibitor (anti-miR). Furthermore, a reasonable causal relation between the miR-10b and HOXD10 expression levels was observed in miR-10b* precursor-transfected cells and miR-10b* inhibitor-transfected cells. The trends of the miRNA alterations obtained using HILO microscopy completely matched the RT-qPCR data and showed remarkable reproducibility (the coefficient of variation [CV] = 0.86%) and sensitivity (<1.0 fM). This proposed imaging method appears to be useful for the simultaneous visualisation of ultra-trace amounts of miRNA and target mRNA and excludes the procedures for RNA extraction and amplification. Therefore, the visualisation of miRNA and the target mRNA should facilitate the exploration of the functions of ultra-trace amounts of miRNA in fixed cells in biological studies and may serve as a powerful tool for diagnoses based on circulating cancer cells.
As-prepared MoS2 quantum dot as a facile fluorescent probe for long-term tracing of live cells
NASA Astrophysics Data System (ADS)
Zhou, Kai; Zhang, Yue; Xia, Zhining; Wei, Weili
2016-07-01
Recently, the newly emerged two-dimensional nanomaterials, layered transition metal dichalcogenide (e.g. MoS2) nanosheets, have drawn tremendous attentions due to their extraordinary electronic and optical properties, and MoS2 quantum dots (MoS2 QDs) with lateral sizes less than 10 nm have been found to be highly luminescent. In the present study, a facile approach for large-scale preparation of MoS2 QDs by Na intercalation reaction without using any toxic organic reagents is proposed. MoS2 QDs were carefully characterized by various techniques including transmission electron microscopy, atomic force microscopy, dynamic light scattering, spectroscopy, in vitro cytotoxicology, and capillary electrophoresis. The as-prepared MoS2 QDs were strongly fluorescent, highly photo-stable, low in cytotoxicity, and readily reactive to thiols. These inherent properties of MoS2 QDs make them excellent fluorescent probes for long-term live cell tracing. The results of live cells imaging indicated that MoS2 QD stained cells remained highly fluorescent after long-term culture, and could be easily traced from other co-cultured cell lines.
Curlik, Daniel M.; DiFeo, Gina; Shors, Tracey J.
2014-01-01
The dentate gyrus of the hippocampal formation generates new granule neurons throughout life. The number of neurons produced each day is inversely related to age, with thousands more produced during puberty than during adulthood, and many fewer produced during senescence. In adulthood, approximately half of these cells undergo apoptosis shortly after they are generated. Most of these cells can be rescued from death by effortful and successful learning experiences (Gould et al., 1999; Waddell and Shors, 2008; Curlik and Shors, 2011). Once rescued, the newly-generated cells differentiate into neurons, and remain in the hippocampus for at least several months (Leuner et al., 2004). Here, we report that many new hippocampal cells also undergo cell death during puberty. Because the juvenile brain is more plastic than during adulthood, and because many experiences are new, we hypothesized that a great number of cells would be rescued by learning during puberty. Indeed, adolescent rats that successfully acquired the trace eyeblink response retained thousands more cells than animals that were not trained, and those that failed to learn. Because the hippocampus generates thousands more cells during puberty than during adulthood, these results support the idea that the adolescent brain is especially responsive to learning. This enhanced response can have significant consequences for the functional integrity of the hippocampus. Such a massive increase in cell proliferation is likely an adaptive response as the young animal must emerge from the care of its mother to face the dangers, challenges, and opportunities of adulthood. PMID:24795549
Thermal phase transition behavior of lipid layers on a single human corneocyte cell.
Imai, Tomohiro; Nakazawa, Hiromitsu; Kato, Satoru
2013-09-01
We have improved the selected area electron diffraction method to analyze the dynamic structural change in a single corneocyte cell non-invasively stripped off from human skin surface. The improved method made it possible to obtain reliable diffraction images to trace the structural change in the intercellular lipid layers on a single corneocyte cell during heating from 24°C to 100°C. Comparison of the results with those of synchrotron X-ray diffraction experiments on human stratum corneum sheets revealed that the intercellular lipid layers on a corneocyte cell exhibit essentially the same thermal phase transitions as those in a stratum corneum sheet. These results suggest that the structural features of the lipid layers are well preserved after the mechanical stripping of the corneocyte cell. Moreover, electron diffraction analyses of the thermal phase transition behaviors of the corneocyte cells that had the lipid layers with different distributions of orthorhombic and hexagonal domains at 24°C suggested that small orthorhombic domains interconnected with surrounding hexagonal domains transforms in a continuous manner into new hexagonal domains. Copyright © 2013 Elsevier Ireland Ltd. All rights reserved.
Evolution of early embryogenesis in rhabditid nematodes
Brauchle, Michael; Kiontke, Karin; MacMenamin, Philip; Fitch, David H. A.; Piano, Fabio
2009-01-01
The cell biological events that guide early embryonic development occur with great precision within species but can be quite diverse across species. How these cellular processes evolve and which molecular components underlie evolutionary changes is poorly understood. To begin to address these questions, we systematically investigated early embryogenesis, from the one- to the four-cell embryo, in 34 nematode species related to C. elegans. We found 40 cell-biological characters that captured the phenotypic differences between these species. By tracing the evolutionary changes on a molecular phylogeny, we found that these characters evolved multiple times and independently of one another. Strikingly, all these phenotypes are mimicked by single-gene RNAi experiments in C. elegans. We use these comparisons to hypothesize the molecular mechanisms underlying the evolutionary changes. For example, we predict that a cell polarity module was altered during the evolution of the Protorhabditis group and show that PAR-1, a kinase localized asymmetrically in C. elegans early embryos, is symmetrically localized in the one-cell stage of Protorhabditis group species. Our genome-wide approach identifies candidate molecules—and thereby modules—associated with evolutionary changes in cell-biological phenotypes. PMID:19643102
Interplay between Short- and Long-Term Plasticity in Cell-Assembly Formation
Hiratani, Naoki; Fukai, Tomoki
2014-01-01
Various hippocampal and neocortical synapses of mammalian brain show both short-term plasticity and long-term plasticity, which are considered to underlie learning and memory by the brain. According to Hebb’s postulate, synaptic plasticity encodes memory traces of past experiences into cell assemblies in cortical circuits. However, it remains unclear how the various forms of long-term and short-term synaptic plasticity cooperatively create and reorganize such cell assemblies. Here, we investigate the mechanism in which the three forms of synaptic plasticity known in cortical circuits, i.e., spike-timing-dependent plasticity (STDP), short-term depression (STD) and homeostatic plasticity, cooperatively generate, retain and reorganize cell assemblies in a recurrent neuronal network model. We show that multiple cell assemblies generated by external stimuli can survive noisy spontaneous network activity for an adequate range of the strength of STD. Furthermore, our model predicts that a symmetric temporal window of STDP, such as observed in dopaminergic modulations on hippocampal neurons, is crucial for the retention and integration of multiple cell assemblies. These results may have implications for the understanding of cortical memory processes. PMID:25007209
Sensitivity studies and laboratory measurements for the laser heterodyne spectrometer experiment
NASA Technical Reports Server (NTRS)
Allario, F.; Katzberg, S. J.; Larsen, J. C.
1980-01-01
Several experiments involving spectral scanning interferometers and gas filter correlation radiometers (ref. 2) using limb scanning solar occultation techniques under development for measurements of stratospheric trace gases from Spacelab and satellite platforms are described. An experiment to measure stratospheric trace constituents by Laser Heterodyne Spectroscopy, a summary of sensitivity analyses, and supporting laboratory measurements are presented for O3, ClO, and H2O2 in which the instrument transfer function is modeled using a detailed optical receiver design.
Role of antioxidants and trace elements in health and immunity of transition dairy cows.
Spears, Jerry W; Weiss, William P
2008-04-01
A number of antioxidants and trace minerals have important roles in immune function and may affect health in transition dairy cows. Vitamin E and beta-carotene are important cellular antioxidants. Selenium (Se) is involved in the antioxidant system via its role in the enzyme glutathione peroxidase. Inadequate dietary vitamin E or Se decreases neutrophil function during the perpariturient period. Supplementation of vitamin E and/or Se has reduced the incidence of mastitis and retained placenta, and reduced duration of clinical symptoms of mastitis in some experiments. Research has indicated that beta-carotene supplementation may enhance immunity and reduce the incidence of retained placenta and metritis in dairy cows. Marginal copper deficiency resulted in reduced neutrophil killing and decreased interferon production by mononuclear cells. Copper supplementation of a diet marginal in copper reduced the peak clinical response during experimental Escherichia coli mastitis. Limited research indicated that chromium supplementation during the transition period may increase immunity and reduce the incidence of retained placenta.
Xiao, Yi; Tholen, Danny; Zhu, Xin-Guang
2016-01-01
Leaf photosynthesis is determined by biochemical properties and anatomical features. Here we developed a three-dimensional leaf model that can be used to evaluate the internal light environment of a leaf and its implications for whole-leaf electron transport rates (J). This model includes (i) the basic components of a leaf, such as the epidermis, palisade and spongy tissues, as well as the physical dimensions and arrangements of cell walls, vacuoles and chloroplasts; and (ii) an efficient forward ray-tracing algorithm, predicting the internal light environment for light of wavelengths between 400 and 2500nm. We studied the influence of leaf anatomy and ambient light on internal light conditions and J. The results show that (i) different chloroplasts can experience drastically different light conditions, even when they are located at the same distance from the leaf surface; (ii) bundle sheath extensions, which are strips of parenchyma, collenchyma or sclerenchyma cells connecting the vascular bundles with the epidermis, can influence photosynthetic light-use efficiency of leaves; and (iii) chloroplast positioning can also influence the light-use efficiency of leaves. Mechanisms underlying leaf internal light heterogeneity and implications of the heterogeneity for photoprotection and for the convexity of the light response curves are discussed. PMID:27702991
Mutant KRAS promotes malignant pleural effusion formation
Αgalioti, Theodora; Giannou, Anastasios D.; Krontira, Anthi C.; Kanellakis, Nikolaos I.; Kati, Danai; Vreka, Malamati; Pepe, Mario; Spella, Μagda; Lilis, Ioannis; Zazara, Dimitra E.; Nikolouli, Eirini; Spiropoulou, Nikolitsa; Papadakis, Andreas; Papadia, Konstantina; Voulgaridis, Apostolos; Harokopos, Vaggelis; Stamou, Panagiota; Meiners, Silke; Eickelberg, Oliver; Snyder, Linda A.; Antimisiaris, Sophia G.; Kardamakis, Dimitrios; Psallidas, Ioannis; Μarazioti, Antonia; Stathopoulos, Georgios T.
2017-01-01
Malignant pleural effusion (MPE) is the lethal consequence of various human cancers metastatic to the pleural cavity. However, the mechanisms responsible for the development of MPE are still obscure. Here we show that mutant KRAS is important for MPE induction in mice. Pleural disseminated, mutant KRAS bearing tumour cells upregulate and systemically release chemokine ligand 2 (CCL2) into the bloodstream to mobilize myeloid cells from the host bone marrow to the pleural space via the spleen. These cells promote MPE formation, as indicated by splenectomy and splenocyte restoration experiments. In addition, KRAS mutations are frequently detected in human MPE and cell lines isolated thereof, but are often lost during automated analyses, as indicated by manual versus automated examination of Sanger sequencing traces. Finally, the novel KRAS inhibitor deltarasin and a monoclonal antibody directed against CCL2 are equally effective against an experimental mouse model of MPE, a result that holds promise for future efficient therapies against the human condition. PMID:28508873
Lemmens, Bennie; van Schendel, Robin; Tijsterman, Marcel
2015-01-01
Faithful DNA replication is vital to prevent disease-causing mutations, chromosomal aberrations and malignant transformation. However, accuracy conflicts with pace and flexibility and cells rely on specialized polymerases and helicases to ensure effective and timely replication of genomes that contain DNA lesions or secondary structures. If and how cells can tolerate a permanent barrier to replication is, however, unknown. Here we show that a single unresolved G-quadruplexed DNA structure can persist through multiple mitotic divisions without changing conformation. Failed replication across a G-quadruplex causes single-strand DNA gaps that give rise to DNA double-strand breaks in subsequent cell divisions, which are processed by polymerase theta (POLQ)-mediated alternative end joining. Lineage tracing experiments further reveal that persistent G-quadruplexes cause genetic heterogeneity during organ development. Our data demonstrate that a single lesion can cause multiple unique genomic rearrangements, and that alternative end joining enables cells to proliferate in the presence of mitotically inherited replication blocks. PMID:26563448
Lemmens, Bennie; van Schendel, Robin; Tijsterman, Marcel
2015-11-13
Faithful DNA replication is vital to prevent disease-causing mutations, chromosomal aberrations and malignant transformation. However, accuracy conflicts with pace and flexibility and cells rely on specialized polymerases and helicases to ensure effective and timely replication of genomes that contain DNA lesions or secondary structures. If and how cells can tolerate a permanent barrier to replication is, however, unknown. Here we show that a single unresolved G-quadruplexed DNA structure can persist through multiple mitotic divisions without changing conformation. Failed replication across a G-quadruplex causes single-strand DNA gaps that give rise to DNA double-strand breaks in subsequent cell divisions, which are processed by polymerase theta (POLQ)-mediated alternative end joining. Lineage tracing experiments further reveal that persistent G-quadruplexes cause genetic heterogeneity during organ development. Our data demonstrate that a single lesion can cause multiple unique genomic rearrangements, and that alternative end joining enables cells to proliferate in the presence of mitotically inherited replication blocks.
Nitrogen fixation system of tungsten-resistant mutants of Azotobacter vinelandii.
Riddle, G D; Simonson, J G; Hales, B J; Braymer, H D
1982-01-01
Mutants of Azotobacter vinelandii ATCC 12837 were isolated which could fix N2 in the presence of high tungsten concentrations. The most studied of these mutants (WD2) grew well in N-free modified Burk broth containing 10 mM W, whereas the wild type would not grow in this medium. WD2 would also grow in Burk N-free broth at about the same rate as the wild type. WD2 in broth containing W exhibited 22% of the whole cell acetylene reduction activity of the wild type in broth containing Mo and showed a lowered affinity for acetylene. Two-dimensional gel electrophoresis experiments showed that N2-fixing cells of WD2 from broth containing W or Mo did not produce significant amounts of component I of native nitrogenase protein. Electron spin resonance spectra of whole cells and cell-free extracts of WD2 from broth containing W lacked any trace of the g = 3.6 resonance associated with FeMoCo. Images PMID:6956567
Osteoblast Production by Reserved Progenitor Cells in Zebrafish Bone Regeneration and Maintenance.
Ando, Kazunori; Shibata, Eri; Hans, Stefan; Brand, Michael; Kawakami, Atsushi
2017-12-04
Mammals cannot re-form heavily damaged bones as in large fracture gaps, whereas zebrafish efficiently regenerate bones even after amputation of appendages. However, the source of osteoblasts that mediate appendage regeneration is controversial. Several studies in zebrafish have shown that osteoblasts are generated by dedifferentiation of existing osteoblasts at injured sites, but other observations suggest that de novo production of osteoblasts also occurs. In this study, we found from cell-lineage tracing and ablation experiments that a group of cells reserved in niches serves as osteoblast progenitor cells (OPCs) and has a significant role in fin ray regeneration. Besides regeneration, OPCs also supply osteoblasts for normal bone maintenance. We further showed that OPCs are derived from embryonic somites, as is the case with embryonic osteoblasts, and are replenished from mesenchymal precursors in adult zebrafish. Our findings reveal that reserved progenitors are a significant and complementary source of osteoblasts for zebrafish bone regeneration. Copyright © 2017 Elsevier Inc. All rights reserved.
Heterogeneity of SOX9 and HNF1β in Pancreatic Ducts Is Dynamic.
Rezanejad, Habib; Ouziel-Yahalom, Limor; Keyzer, Charlotte A; Sullivan, Brooke A; Hollister-Lock, Jennifer; Li, Wan-Chun; Guo, Lili; Deng, Shaopeng; Lei, Ji; Markmann, James; Bonner-Weir, Susan
2018-03-13
Pancreatic duct epithelial cells have been suggested as a source of progenitors for pancreatic growth and regeneration. However, genetic lineage-tracing experiments with pancreatic duct-specific Cre expression have given conflicting results. Using immunofluorescence and flow cytometry, we show heterogeneous expression of both HNF1β and SOX9 in adult human and murine ductal epithelium. Their expression was dynamic and diminished significantly after induced replication. Purified pancreatic duct cells formed organoid structures in 3D culture, and heterogeneity of expression of Hnf1β and Sox9 was maintained even after passaging. Using antibodies against a second cell surface molecule CD51 (human) or CD24 (mouse), we could isolate living subpopulations of duct cells enriched for high or low expression of HNF1β and SOX9. Only the CD24 high (Hnfβ high /Sox9 high ) subpopulation was able to form organoids. Copyright © 2018 The Author(s). Published by Elsevier Inc. All rights reserved.
Profilin1 activity in cerebellar granule neurons is required for radial migration in vivo
Kullmann, Jan A; Wickertsheim, Ines; Minnerup, Lara; Costell, Mercedes; Friauf, Eckhard; Rust, Marco B
2015-01-01
Neuron migration defects are an important aspect of human neuropathies. The underlying molecular mechanisms of such migration defects are largely unknown. Actin dynamics has been recognized as an important determinant of neuronal migration, and we recently found that the actin-binding protein profilin1 is relevant for radial migration of cerebellar granule neurons (CGN). As the exploited brain-specific mutants lacked profilin1 in both neurons and glial cells, it remained unknown whether profilin1 activity in CGN is relevant for CGN migration in vivo. To test this, we capitalized on a transgenic mouse line that expresses a tamoxifen-inducible Cre variant in CGN, but no other cerebellar cell type. In these profilin1 mutants, the cell density was elevated in the molecular layer, and ectopic CGN occurred. Moreover, 5-bromo-2′-deoxyuridine tracing experiments revealed impaired CGN radial migration. Hence, our data demonstrate the cell autonomous role of profilin1 activity in CGN for radial migration. PMID:25495756
Image feature based GPS trace filtering for road network generation and road segmentation
Yuan, Jiangye; Cheriyadat, Anil M.
2015-10-19
We propose a new method to infer road networks from GPS trace data and accurately segment road regions in high-resolution aerial images. Unlike previous efforts that rely on GPS traces alone, we exploit image features to infer road networks from noisy trace data. The inferred road network is used to guide road segmentation. We show that the number of image segments spanned by the traces and the trace orientation validated with image features are important attributes for identifying GPS traces on road regions. Based on filtered traces , we construct road networks and integrate them with image features to segmentmore » road regions. Lastly, our experiments show that the proposed method produces more accurate road networks than the leading method that uses GPS traces alone, and also achieves high accuracy in segmenting road regions even with very noisy GPS data.« less
Image feature based GPS trace filtering for road network generation and road segmentation
DOE Office of Scientific and Technical Information (OSTI.GOV)
Yuan, Jiangye; Cheriyadat, Anil M.
We propose a new method to infer road networks from GPS trace data and accurately segment road regions in high-resolution aerial images. Unlike previous efforts that rely on GPS traces alone, we exploit image features to infer road networks from noisy trace data. The inferred road network is used to guide road segmentation. We show that the number of image segments spanned by the traces and the trace orientation validated with image features are important attributes for identifying GPS traces on road regions. Based on filtered traces , we construct road networks and integrate them with image features to segmentmore » road regions. Lastly, our experiments show that the proposed method produces more accurate road networks than the leading method that uses GPS traces alone, and also achieves high accuracy in segmenting road regions even with very noisy GPS data.« less
Investigating the potential for subsurface primary production fueled by serpentinization
NASA Astrophysics Data System (ADS)
Brazelton, W. J.; Nelson, B. Y.; Schrenk, M. O.
2011-12-01
Ultramafic rocks in the Earth's mantle represent a tremendous reservoir of carbon and reducing power. Tectonic uplift of these materials into the crust can result in serpentinization, a highly exothermic geochemical reaction that releases hydrogen gas (H2) and promotes the abiogenic synthesis of organic molecules. The extent and activity of microbial communities in serpentinite-hosted subsurface habitats is almost entirely unknown, but they clearly have great potential to host extensive sunlight-independent primary production fueled by H2 and abiotic carbon compounds. We have been testing this hypothesis at several sites of serpentinization around the globe utilizing a suite of techniques including metagenomics, 16S rRNA pyrotag sequencing, and stable isotope tracing experiments. All four of our study sites, which include deep-sea hydrothermal vents, terrestrial alkaline springs, and continental drill holes, are characteristically low in archaeal and bacterial genetic diversity. In carbonate chimneys of the Lost City hydrothermal field (Mid-Atlantic Ridge), for example, a single archaeal phylotype dominates the biofilm community. Stable isotope tracing experiments indicated that these archaeal biofilms are capable of both production and anaerobic oxidation of methane at 80C and pH 10. Both production and oxidation were stimulated by H2, suggesting a possible syntrophic relationship among cells within the biofilm. Preliminary results from similar stable isotope tracing experiments at terrestrial alkaline seeps at the Tablelands Ophiolite (Newfoundland), Ligurian springs (Italy), and McLaughlin Reserve (California) have indicated the potential for microbial activity fueled by H2 and acetate. Furthermore, recent metagenomic sequencing of fluids from the Tablelands and Ligurian springs have revealed genomic potential for chemolithotrophy powered by iron reduction with H2. In summary, these data support the potential for extensive microbial activity fueled by serpentinization, and further characterization of the endemic organisms (now underway) is likely to reveal novel physiological adaptations to the unusual conditions of serpentinite-hosted habitats.
Choi, Tae-Young; Ninov, Nikolay; Stainier, Didier Y R; Shin, Donghun
2014-03-01
Biliary epithelial cells (BECs) are considered to be a source of regenerating hepatocytes when hepatocyte proliferation is compromised. However, there is still controversy about the extent to which BECs can contribute to the regenerating hepatocyte population, and thereby to liver recovery. To investigate this issue, we established a zebrafish model of liver regeneration in which the extent of hepatocyte ablation can be controlled. Hepatocytes were depleted by administration of metronidazole to Tg(fabp10a:CFP-NTR) animals. We traced the origin of regenerating hepatocytes using short-term lineage-tracing experiments, as well as the inducible Cre/loxP system; specifically, we utilized both a BEC tracer line Tg(Tp1:CreER(T2)) and a hepatocyte tracer line Tg(fabp10a:CreER(T2)). We also examined BEC and hepatocyte proliferation and liver marker gene expression during liver regeneration. BECs gave rise to most of the regenerating hepatocytes in larval and adult zebrafish after severe hepatocyte depletion. After hepatocyte loss, BECs proliferated as they dedifferentiated into hepatoblast-like cells; they subsequently differentiated into highly proliferative hepatocytes that restored the liver mass. This process was impaired in zebrafish wnt2bb mutants; in these animals, hepatocytes regenerated but their proliferation was greatly reduced. BECs contribute to regenerating hepatocytes after substantial hepatocyte depletion in zebrafish, thereby leading to recovery from severe liver damage. Copyright © 2014 AGA Institute. Published by Elsevier Inc. All rights reserved.
A Design Basis for Spacecraft Cabin Trace Contaminant Control
NASA Technical Reports Server (NTRS)
Perry, Jay L.
2009-01-01
Successful trace chemical contamination control is one of the components necessary for achieving good cabin atmospheric quality. While employing seemingly simple process technologies, sizing the active contamination control equipment must employ a reliable design basis for the trace chemical load in the cabin atmosphere. A simplified design basis that draws on experience gained from the International Space Station program is presented. The trace chemical contamination control design load refines generation source magnitudes and includes key chemical functional groups representing both engineering and toxicology challenges.
Terada, Maiko; Horisawa, Kenichi; Miura, Shizuka; Takashima, Yasuo; Ohkawa, Yasuyuki; Sekiya, Sayaka; Matsuda-Ito, Kanae; Suzuki, Atsushi
2016-01-01
Intrahepatic cholangiocarcinoma (ICC) is a malignant epithelial neoplasm composed of cells resembling cholangiocytes that line the intrahepatic bile ducts in portal areas of the hepatic lobule. Although ICC has been defined as a tumor arising from cholangiocyte transformation, recent evidence from genetic lineage-tracing experiments has indicated that hepatocytes can be a cellular origin of ICC by directly changing their fate to that of biliary lineage cells. Notch signaling has been identified as an essential factor for hepatocyte conversion into biliary lineage cells at the onset of ICC. However, the mechanisms underlying Notch signal activation in hepatocytes remain unclear. Here, using a mouse model of ICC, we found that hepatic macrophages called Kupffer cells transiently congregate around the central veins in the liver and express the Notch ligand Jagged-1 coincident with Notch activation in pericentral hepatocytes. Depletion of Kupffer cells prevents the Notch-mediated cell-fate conversion of hepatocytes to biliary lineage cells, inducing hepatocyte apoptosis and increasing mortality in mice. These findings will be useful for uncovering the pathogenic mechanism of ICC and developing prevenient and therapeutic strategies for this refractory disease. PMID:27698452
NASA Astrophysics Data System (ADS)
Fietz, S.; Roychoudhury, A. N.; Thomalla, S.; Mtshali, T. N.; Philibert, R.; Van Horsten, N.; Loock, J. C.; Cloete, R.
2016-02-01
Phytoplankton primary productivity depends on macro- and micronutrient availability and in turn plays a key role in the marine biogeochemical cycles. The role of iron in regulating phytoplankton primary production and thus biogeochemical cycles in the Southern Ocean has been widely recognized; however, it also became obvious that iron is not the sole factor limiting primary production in the Southern Ocean and that light, for instance, might aggravate or relief trace nutrient limitation. We conducted a suite of ship-board incubation experiments in austral summer 2013/14, 2014/15 and winter 2015 to shed light on the complex interplay between trace metal and light limitation. We observed a strong difference in acclimation and photophysiological response depending on the environmental conditions of the in-situ communities prior to the experiment. The differences in acclimation and photophysiological responses resulted in different growth and macronutrient uptake rates. Revisited stations did, however, not always show the same responses. At at least one station we will link the incubation experiments to the in-situ vertical profiles of trace metals, macronutrients and primary productivity.
Parabolic Flight Evaluation of Bacterial Adhesion on Multiple Antimicrobial Surface Treatments
NASA Technical Reports Server (NTRS)
Birmele, Michele
2011-01-01
This report describes the development of a test method and the evaluation of the effectiveness of antimicrobial technologies in reduced gravity based on parabolic flight experiments. Microbial growth is a common occurrence on fully immersed wetted surfaces in spacecraft environmental control and life support systems despite the use of chemical and/or physical \\disinfection. Many materials and surface treatments with antimicrobial properties are commercially available but none have been vetted for spaceflight applications. Herein a test method is explained that included ground and reduced gravity parabolic flight experiments with a standard microorganism recovered from spacecraft, Pseudomonas aeruginosa, added at a concentration of 1 x 10(exp 5) cells per milliliter (mL) onto challenge material coupon surfaces. Several experimental materials were observed to slightly reduce microbial attachment in reduced gravity flight experiments, but none were capable of eliminating all challenge bacteria. Lunar gravity had an increased antimicrobial effect in 28 out of 36 test coupons compared to microgravity when provided otherwise identical conditions for growth, suggesting trace .amounts of gravity may be required for maximum antimicrobial performance. Bacterial cells exposed to variable gravity had more than twice as ,much intracellular adenosine triphosphate (ATP) when compared to control cells exposed only to Earth gravity due to a short duration response to environmental stress. An ATP luminescence assay was the method most amenable to development of an in-flight microbial monitoring assay
NASA Technical Reports Server (NTRS)
Dyroff, Christoph; Fried, Alan; Richter, Dirk; Walega, James G.; Zahniser, Mark S.; McManus, J. Barry
2005-01-01
The present paper discusses a new, more stable, astigmatic Herriott cell employing carbon fiber stabilizing rods. Laboratory tests using a near-IR absorption feature of CO at 1564.168-nm revealed a factor of two improvement in measurement stability compared with the present commercial design when the sampling pressure was changed by +/-2 Torr around 50 Torr. This new cell should significantly enhance our efforts to measure trace gases employing pathlengths of 100 to 200-meters on airborne platforms with minimum detectable line center absorbances of less than 10(exp -6).
Tasaki, Junichi; Uchiyama-Tasaki, Chihiro; Rouhana, Labib
2016-01-01
Planarian flatworms have become an important system for the study of stem cell behavior and regulation in vivo. These organisms are able to regenerate any part of their body upon damage or amputation. A crucial cellular event in the process of planarian regeneration is the migration of pluripotent stem cells (known as neoblasts) to the site of injury. Here we describe two approaches for analyzing migration of planarian stem cells to an area where these have been ablated by localized X-ray irradiation. The first approach involves immunolabeling of mitotic neoblasts, while the second is based on tracing stem cells and their progeny after BrdU incorporation. The use of planarians in studies of cell motility is suitable for the identification of factors that influence stem cell migration in vivo and is amenable to RNA interference or pharmacological screening.
Concepts of Cell Lineage in Mammalian Embryos.
Papaioannou, Virginia E
2016-01-01
Cell lineage is the framework for understanding cellular diversity, stability of differentiation, and its relationship to pluripotency. The special condition of in utero development in mammals has presented challenges to developmental biologists in tracing cell lineages but modern imaging and cell marking techniques have allowed the gradual elucidation of lineage relationships. Early experimental embryology approaches had limited resolution and relied of suboptimal cell markers and considerable disturbance to the embryos. Transgenic technology introduced genetic markers, particularly fluorescent proteins that, combined with sophisticated imaging modalities, greatly increase resolution and allow clonal analysis within lineages. The concept of cell lineage has also undergone evolution as it became possible to trace the lineage of cells based not only on their physical location or attributes but also on their gene expression pattern, thus opening up mechanistic lines of investigation into the determinants of cell lineage. © 2016 Elsevier Inc. All rights reserved.
Germ stem cells are active in postnatal mouse ovary under physiological conditions
Guo, Kun; Li, Chao-hui; Wang, Xin-yi; He, Da-jian; Zheng, Ping
2016-01-01
STUDY HYPOTHESIS Are active ovarian germ stem cells present in postnatal mouse ovaries under physiological conditions? STUDY FINDING Active ovarian germ stem cells exist and function in adult mouse ovaries under physiological conditions. WHAT IS KNOWN ALREADY In vitro studies suggested the existence of germ stem cells in postnatal ovaries of mouse, pig and human. However, in vivo studies provided evidence against the existence of active germ stem cells in postnatal mouse ovaries. Thus, it remains controversial whether such germ stem cells really exist and function in vivo in postnatal mammalian ovaries. STUDY DESIGN, SAMPLES/MATERIALS, METHODS Octamer-binding transcription factor 4 (Oct4)-MerCreMer transgenic mice were crossed with R26R-enhanced yellow fluorescent protein (EYFP) mice to establish a tamoxifen-inducible tracing system so that Oct4-expressing potential ovarian germ stem cells in young adult mice (5–6 weeks old) can be labeled with EYFP. The germ cell activities of DNA replication, mitotic division, entry into meiosis and progression to primordial follicle stage were investigated by means of immunofluorescent staining of ovarian tissues collected at different time points post-tamoxifen injection (1 day, 3 days, 2 months and 4 months). Meiosis entry and primordial follicle formation were also measured by EYFP-labeled single-cell RT–PCR. Germ cell proliferation and mitotic division were examined through 5-bromodeoxyuridine triphosphate incorporation assay. At each time point, ovaries from two to three animals were used for each set of experiment. MAIN RESULTS AND THE ROLE OF CHANCE By labeling the Oct4-expressing small germ cells and tracing their fates for up to 4 months, we observed persistent meiosis entry and primordial follicle replenishment. Furthermore, we captured the transient processes of mitotic DNA replication as well as mitotic division of the marked germ cells at various time periods after tracing. These lines of evidence unambiguously support the presence of active germ stem cells in postnatal ovaries and their function in replenishing primordial follicle pool under physiological conditions. Moreover, we pointed out that Oct4+ deleted in azoospermia-like (Dazl)− but not Oct4+Dazl+ or Oct4+ DEAD (Asp–Glu–Ala–Asp) Box Polypeptide 4 (Ddx4)+ cells contain a population of germ stem cells in mouse ovary. LIMITATIONS, REASONS FOR CAUTION This study was conducted in mice. Whether or not the results are applicable to human remain unclear. The future work should aim at identifying the specific ovarian germ stem cell marker and evaluating the significance of these stem cells to normal ovarian function. WIDER IMPLICATIONS OF THE FINDINGS Clarifying the existence of active germ stem cells and their functional significance in postnatal mammalian ovaries could provide new insights in understanding the mechanism of ovarian aging and failure. LARGE SCALE DATA Not applicable. STUDY FUNDING/COMPETING INTEREST(S) This work was supported by the National Key Basic Research Program of China (grant number 2012CBA01300) and the National Natural Science Foundation of China to P.Z. (31571484). No competing interests are reported. PMID:26916381
Feist, Maren; Schwarzfischer, Philipp; Heinrich, Paul; Sun, Xueni; Kemper, Judith; von Bonin, Frederike; Perez-Rubio, Paula; Taruttis, Franziska; Rehberg, Thorsten; Dettmer, Katja; Gronwald, Wolfram; Reinders, Jörg; Engelmann, Julia C; Dudek, Jan; Klapper, Wolfram; Trümper, Lorenz; Spang, Rainer; Oefner, Peter J; Kube, Dieter
2018-04-17
Knowledge of stromal factors that have a role in the transcriptional regulation of metabolic pathways aside from c-Myc is fundamental to improvements in lymphoma therapy. Using a MYC-inducible human B-cell line, we observed the cooperative activation of STAT3 and NF-κB by IL10 and CpG stimulation. We show that IL10 + CpG-mediated cell proliferation of MYC low cells depends on glutaminolysis. By 13 C- and 15 N-tracing of glutamine metabolism and metabolite rescue experiments, we demonstrate that GOT2 provides aspartate and nucleotides to cells with activated or aberrant Jak/STAT and NF-κB signaling. A model of GOT2 transcriptional regulation is proposed, in which the cooperative phosphorylation of STAT3 and direct joint binding of STAT3 and p65/NF-κB to the proximal GOT2 promoter are important. Furthermore, high aberrant GOT2 expression is prognostic in diffuse large B-cell lymphoma underscoring the current findings and importance of stromal factors in lymphoma biology.
Yin, Hang; Pasut, Alessandra; Soleimani, Vahab D; Bentzinger, C Florian; Antoun, Ghadi; Thorn, Stephanie; Seale, Patrick; Fernando, Pasan; van Ijcken, Wilfred; Grosveld, Frank; Dekemp, Robert A; Boushel, Robert; Harper, Mary-Ellen; Rudnicki, Michael A
2013-02-05
Brown adipose tissue (BAT) is an energy-dispensing thermogenic tissue that plays an important role in balancing energy metabolism. Lineage-tracing experiments indicate that brown adipocytes are derived from myogenic progenitors during embryonic development. However, adult skeletal muscle stem cells (satellite cells) have long been considered uniformly determined toward the myogenic lineage. Here, we report that adult satellite cells give rise to brown adipocytes and that microRNA-133 regulates the choice between myogenic and brown adipose determination by targeting the 3'UTR of Prdm16. Antagonism of microRNA-133 during muscle regeneration increases uncoupled respiration, glucose uptake, and thermogenesis in local treated muscle and augments whole-body energy expenditure, improves glucose tolerance, and impedes the development of diet-induced obesity. Finally, we demonstrate that miR-133 levels are downregulated in mice exposed to cold, resulting in de novo generation of satellite cell-derived brown adipocytes. Therefore, microRNA-133 represents an important therapeutic target for the treatment of obesity. Copyright © 2013 Elsevier Inc. All rights reserved.
Cell tracking supports secondary gastrulation in the moon jellyfish Aurelia.
Gold, David A; Nakanishi, Nagayasu; Hensley, Nicholai M; Hartenstein, Volker; Jacobs, David K
2016-11-01
The moon jellyfish Aurelia exhibits a dramatic reorganization of tissue during its metamorphosis from planula larva to polyp. There are currently two competing hypotheses regarding the fate of embryonic germ layers during this metamorphosis. In one scenario, the original endoderm undergoes apoptosis and is replaced by a secondary endoderm derived from ectodermal cells. In the second scenario, both ectoderm and endoderm remain intact through development. In this study, we performed a pulse-chase experiment to trace the fate of larval ectodermal cells. We observed that prior to metamorphosis, ectodermal cells that proliferated early in larval development concentrate at the future oral end of the polyp. During metamorphosis, these cells migrate into the endoderm, extending all the way to the aboral portion of the gut. We therefore reject the hypothesis that larval endoderm remains intact during metamorphosis and provide additional support for the "secondary gastrulation" hypothesis. Aurelia appears to offer the first and only described case where a cnidarian derives its endoderm twice during normal development, adding to a growing body of evidence that germ layers can be dramatically reorganized in cnidarian life cycles.
Tracing the origin of disseminated tumor cells in breast cancer using single-cell sequencing.
Demeulemeester, Jonas; Kumar, Parveen; Møller, Elen K; Nord, Silje; Wedge, David C; Peterson, April; Mathiesen, Randi R; Fjelldal, Renathe; Zamani Esteki, Masoud; Theunis, Koen; Fernandez Gallardo, Elia; Grundstad, A Jason; Borgen, Elin; Baumbusch, Lars O; Børresen-Dale, Anne-Lise; White, Kevin P; Kristensen, Vessela N; Van Loo, Peter; Voet, Thierry; Naume, Bjørn
2016-12-09
Single-cell micro-metastases of solid tumors often occur in the bone marrow. These disseminated tumor cells (DTCs) may resist therapy and lay dormant or progress to cause overt bone and visceral metastases. The molecular nature of DTCs remains elusive, as well as when and from where in the tumor they originate. Here, we apply single-cell sequencing to identify and trace the origin of DTCs in breast cancer. We sequence the genomes of 63 single cells isolated from six non-metastatic breast cancer patients. By comparing the cells' DNA copy number aberration (CNA) landscapes with those of the primary tumors and lymph node metastasis, we establish that 53% of the single cells morphologically classified as tumor cells are DTCs disseminating from the observed tumor. The remaining cells represent either non-aberrant "normal" cells or "aberrant cells of unknown origin" that have CNA landscapes discordant from the tumor. Further analyses suggest that the prevalence of aberrant cells of unknown origin is age-dependent and that at least a subset is hematopoietic in origin. Evolutionary reconstruction analysis of bulk tumor and DTC genomes enables ordering of CNA events in molecular pseudo-time and traced the origin of the DTCs to either the main tumor clone, primary tumor subclones, or subclones in an axillary lymph node metastasis. Single-cell sequencing of bone marrow epithelial-like cells, in parallel with intra-tumor genetic heterogeneity profiling from bulk DNA, is a powerful approach to identify and study DTCs, yielding insight into metastatic processes. A heterogeneous population of CNA-positive cells is present in the bone marrow of non-metastatic breast cancer patients, only part of which are derived from the observed tumor lineages.
Scaling properties of ballistic nano-transistors
2011-01-01
Recently, we have suggested a scale-invariant model for a nano-transistor. In agreement with experiments a close-to-linear thresh-old trace was found in the calculated ID - VD-traces separating the regimes of classically allowed transport and tunneling transport. In this conference contribution, the relevant physical quantities in our model and its range of applicability are discussed in more detail. Extending the temperature range of our studies it is shown that a close-to-linear thresh-old trace results at room temperatures as well. In qualitative agreement with the experiments the ID - VG-traces for small drain voltages show thermally activated transport below the threshold gate voltage. In contrast, at large drain voltages the gate-voltage dependence is weaker. As can be expected in our relatively simple model, the theoretical drain current is larger than the experimental one by a little less than a decade. PMID:21711899
Viking and Mars Rover exobiology
NASA Technical Reports Server (NTRS)
Schwartz, D. E.; Mancinelli, Rocco L.; Ohara, B. J.
1989-01-01
Other than Earth, Mars is the planet generating the greatest interest among those researching and contemplating the origin and distribution of life throughout the universe. The similarity of the early environments of Earth and Mars, and the biological evolution on early Earth provides the motivation to seriously consider the possibility of a primordial Martian biosphere. In 1975 the Viking project launched two unmanned spacecraft to Mars with the intent of finding evidence of the existence of present or past life on this planet. Three Viking Biology experiments were employed: the Labeled Release experiment, the Gas Exchange Experiment, and the Pyrolytic Release experiment. Each of these three experiments tested for microbial existence and utilization of a substrate by examining the gases evolved from specific chemical reactions. Although the results of these experiments were inconclusive, they inferred that there are no traces of extant life on Mars. However, the experiments did not specifically look for indication of extinct life. Therefore, most of the exobiologic strategies and experiments suggested for the Mars Rover Sample Return Mission involve searching for signature of extinct life. The most significant biological signatures and chemical traces to detect include: isotopic and chemical signatures of metabolic activity, anomalous concentrations of certain metals, trace and microfossils, organically preserved materials, carbonates, nitrates, and evaporites.
NASA Technical Reports Server (NTRS)
Chiou, E. W.; Mccormick, M. P.; Mcmaster, L. R.; Chu, W. P.; Larsen, J. C.; Rind, D.; Oltmans, S.
1993-01-01
A comparison is made of the stratospheric water vapor measurements made by the satellite sensors of the Stratospheric Aerosol and Gas Experiment II (SAGE II), the Nimbus-7 LIMS, and the Spacelab 3 Atmospheric Trace Molecule Spectroscopy (ATMOS) experiment. It was found that, despite differences in the measurement techniques, sampling bias, and observational periods, the three experiments have disclosed a generally consistent pattern of stratospheric water vapor distribution. The only significant difference occurs at high southern altitudes in May below 18 km, where LIMS measurements were 2-3 ppmv greater than those of SAGE II and ATMOS.
Estimation of Mineral and Trace Element Profile in Bubaline Milk Affected with Subclinical Mastitis.
Singh, Mahavir; Yadav, Poonam; Sharma, Anshu; Garg, V K; Mittal, Dinesh
2017-04-01
The milk samples from buffaloes of Murrah breed at mid lactation stage, reared at an organised dairy farm, were screened for subclinical mastitis based on bacteriological examination and somatic cell count following International Dairy Federation criteria. Milk samples from subclinical mastitis infected and healthy buffaloes were analysed to evaluate physicochemical alterations in terms of protein, fat, pH, electrical conductivity, chloride, minerals (sodium, potassium and calcium) and trace elements (iron, zinc, copper and selenium). In the present study, protein, fat, zinc, iron, calcium and selenium content was significantly lower (P < 0.001), while pH and electrical conductivity were significantly higher in mastitic milk as compared to normal milk. Concentration of electrolytes mainly sodium and chloride significantly increased with higher somatic cell count in mastitic milk and to maintain osmolality; potassium levels decreased proportionately. Correlation matrix revealed significantly positive interdependences of somatic cell count with pH, electrical conductivity, sodium and chloride. However, protein, fat, calcium and potassium were correlated negatively with elevated somatic cell count in mastitic milk. It is concluded that udder infections resulting in elevated somatic cells may alter the mineral and trace element profile of milk, and magnitude of changes may have diagnostic and prognostic value.
Laser heating of gold nanoparticles: photothermal cancer cell therapy
NASA Astrophysics Data System (ADS)
Nedyalkov, N. N.; Atanasov, P. A.; Toshkova, R. A.; Gardeva, E. G.; Yossifova, L. S.; Alexandrov, M. T.; Karashanova, D.
2012-06-01
In this work an application of gold nanoparticles in in-vitro photothermal cancer cell therapy is demonstrated. Gold nanoparticles with different diameters - 40, 100 and 200 nm are mixed with HeLa cancer cells. After incubation, the nanoparticles are found to be deposited on the cell's membrane or enter into the cells. Pulsed laser radiation at wavelength of 532 nm delivered by Nd:YAG system is used to irradiate the samples. The experiments are performed at fluences in the range from 50 mJ/cm2 up to the established safety standard for medical lasers of 100 mJ/cm2. The cell viability as a function of the particle dimensions and laser fluence is estimated. The nanoparticles heating and cooling dynamics is traced by a numerical model based on heat diffusion equation combined with Mie theory for calculation of the optical properties of nanoparticles. The particle response to the nanosecond laser heating is investigated experimentally as gold colloids are irradiated at different fluences. The threshold fluences for particle's melting and boiling are defined. We show that at the presented fluence range the particles are decomposed into smaller fragments and even short irradiation time leads to decrease of cell viability.
ERIC Educational Resources Information Center
Spudich, Thomas M.; Herrmann, Jennifer K.; Fietkau, Ronald; Edwards, Grant A.
2004-01-01
An experiment is conducted to ascertain trace-level Pb in samples of bovine liver or muscle by applying graphite furnace atomic absorption spectrophotometry (GFAAS). The primary objective is to display the effects of physical and spectral intrusions in determining trace elements, and project the usual methods employed to minimize accuracy errors…
Satoh, Kanehide; Kominami, Tetsuya
2008-10-01
To elucidate factors involved in the oral-aboral axis specification, several observations and experiments were undertaken using the sand dollar Scaphechinus mirabilis. Unlike in Strongylcentrotus purpuratus, localization of mitochondria was not detected in unfertilized eggs. After fertilization, however, the bulk of mitochondria became localized to the opposite side of sperm entry. The first cleavage divided this mitochondrial cluster into daughter blastomeres. On the other hand, a second cleavage produced daughter blastomeres containing quite different amounts of mitochondria. To know whether such mitochondrial localization affects the oral-aboral axis specification, 4-cell-stage embryos were separated along the second cleavage plane. Although both half embryos developed into morphologically normal plutei, some differences, such as the number of pigment cells, were noticed between the siblings. In contrast, cell tracing revealed that the first cleavage separated the oral from the aboral part in most cases, indicating that the unequal distribution of mitochondria is not critical for the oral-aboral axis specification. Further, stained and non-stained half embryo fragments were combined. Such combined embryos developed into normal plutei with a single oral-aboral axis. The plane dividing labeled and non-labeled parts were incident, oblique or perpendicular to the median plane of the combined embryo, and the appearance frequencies of those labeling patterns were similar to those obtained by cell tracing in intact embryos. Interestingly, the half fragments derived from embryos inseminated earlier showed a tendency to form the oral part. These suggest that several factors as well as the localized cytoplasmic components would be involved in the specification process of oral-aboral axis.
An R package for state-trace analysis.
Prince, Melissa; Hawkins, Guy; Love, Jonathon; Heathcote, Andrew
2012-09-01
State-trace analysis (Bamber, Journal of Mathematical Psychology, 19, 137-181, 1979) is a graphical analysis that can determine whether one or more than one latent variable mediates an apparent dissociation between the effects of two experimental manipulations. State-trace analysis makes only ordinal assumptions and so, is not confounded by range effects that plague alternative methods, especially when performance is measured on a bounded scale (such as accuracy). We describe and illustrate the application of a freely available GUI driven package, StateTrace, for the R language. StateTrace automates many aspects of a state-trace analysis of accuracy and other binary response data, including customizable graphics and the efficient management of computationally intensive Bayesian methods for quantifying evidence about the outcomes of a state-trace experiment, developed by Prince, Brown, and Heathcote (Psychological Methods, 17, 78-99, 2012).
NASA Technical Reports Server (NTRS)
Kleb, Mary M.; Scott, A. Donald, Jr.
2003-01-01
This report provides a compendium of NASA aircraft data that are available from NASA's Global Tropospheric Experiment's (GTE) Transport and Chemical Evolution over the Pacific (TRACE-P) Mission. The broad goal of TRACE-P was to characterize the transit and evolution of the Asian outflow over the western Pacific. Conducted from February 24 through April 10, 2001, TRACE-P integrated airborne, satellite- and ground based observations, as well as forecasts from aerosol and chemistry models. The format of this compendium utilizes data plots (time series) of selected data acquired aboard the NASA/Dryden DC-8 (vol. 1) and NASA/Wallops P-3B (vol. 2) aircraft during TRACE-P. The purpose of this document is to provide a representation of aircraft data that are available in archived format via NASA Langley's Distributed Active Archive Center (DAAC) and through the GTE Project Office archive. The data format is not intended to support original research/analyses, but to assist the reader in identifying data that are of interest.
NASA Technical Reports Server (NTRS)
Kleb, Mary M.; Scott, A. Donald, Jr.
2003-01-01
This report provides a compendium of NASA aircraft data that are available from NASA's Global Tropospheric Experiment's (GTE) Transport and Chemical Evolution over the Pacific (TRACE-P) Mission. The broad goal of TRACE-P was to characterize the transit and evolution of the Asian outflow over the western Pacific. Conducted from February 24 through April 10, 2001, TRACE-P integrated airborne, satellite- and ground-based observations, as well as forecasts from aerosol and chemistry models. The format of this compendium utilizes data plots (time series) of selected data acquired aboard the NASA/Dryden DC-8 (vol. 1) and NASA/Wallops P-3B (vol. 2) aircraft during TRACE-P. The purpose of this document is to provide a representation of aircraft data that are available in archived format via NASA Langley s Distributed Active Archive Center (DAAC) and through the GTE Project Office archive. The data format is not intended to support original research/analyses, but to assist the reader in identifying data that are of interest.
Mammary stem cells and the differentiation hierarchy: current status and perspectives
Visvader, Jane E.; Stingl, John
2014-01-01
The mammary epithelium is highly responsive to local and systemic signals, which orchestrate morphogenesis of the ductal tree during puberty and pregnancy. Based on transplantation and lineage tracing studies, a hierarchy of stem and progenitor cells has been shown to exist among the mammary epithelium. Lineage tracing has highlighted the existence of bipotent mammary stem cells (MaSCs) in situ as well as long-lived unipotent cells that drive morphogenesis and homeostasis of the ductal tree. Moreover, there is accumulating evidence for a heterogeneous MaSC compartment comprising fetal MaSCs, slow-cycling cells, and both long-term and short-term repopulating cells. In parallel, diverse luminal progenitor subtypes have been identified in mouse and human mammary tissue. Elucidation of the normal cellular hierarchy is an important step toward understanding the “cells of origin” and molecular perturbations that drive breast cancer. PMID:24888586
Zinc in innate and adaptive tumor immunity
2010-01-01
Zinc is important. It is the second most abundant trace metal with 2-4 grams in humans. It is an essential trace element, critical for cell growth, development and differentiation, DNA synthesis, RNA transcription, cell division, and cell activation. Zinc deficiency has adverse consequences during embryogenesis and early childhood development, particularly on immune functioning. It is essential in members of all enzyme classes, including over 300 signaling molecules and transcription factors. Free zinc in immune and tumor cells is regulated by 14 distinct zinc importers (ZIP) and transporters (ZNT1-8). Zinc depletion induces cell death via apoptosis (or necrosis if apoptotic pathways are blocked) while sufficient zinc levels allows maintenance of autophagy. Cancer cells have upregulated zinc importers, and frequently increased zinc levels, which allow them to survive. Based on this novel synthesis, approaches which locally regulate zinc levels to promote survival of immune cells and/or induce tumor apoptosis are in order. PMID:21087493
Ray tracing through a hexahedral mesh in HADES
DOE Office of Scientific and Technical Information (OSTI.GOV)
Henderson, G L; Aufderheide, M B
In this paper we describe a new ray tracing method targeted for inclusion in HADES. The algorithm tracks rays through three-dimensional tetrakis hexahedral mesh objects, like those used by the ARES code to model inertial confinement experiments.
Trace Elements: The Little Things that Count
ERIC Educational Resources Information Center
Evans, Gary W.
1977-01-01
Examines recent findings regarding the importance of trace elements to good health. Copper, chromium, and zinc are discussed. Described also are multi-disciplinary experiments in progress at the Human Nutrition Laboratory, a federal facility in Grand Forks, North Dakota. (CS)
Lgr5-EGFP marks taste bud stem/progenitor cells in posterior tongue
Yee, Karen K.; Li, Yan; Redding, Kevin M.; Iwatsuki, Ken; Margolskee, Robert F.; Jiang, Peihua
2013-01-01
Until recently, reliable markers for adult stem cells have been lacking for many regenerative mammalian tissues. Lgr5 (leucine-rich repeat-containing G-protein coupled receptor 5) has been identified as a marker for adult stem cells in intestine, stomach, and hair follicle; Lgr5-expressing cells give rise to all types of cells in these tissues. Taste epithelium also regenerates constantly, yet the identity of adult taste stem cells remains elusive. In this study, we found that Lgr5 is strongly expressed in cells at the bottom of trench areas at the base of circumvallate and foliate taste papillae and weakly expressed in the basal area of taste buds and that Lgr5-expressing cells in posterior tongue are a subset of K14-positive epithelial cells. Lineage-tracing experiments using an inducible Cre knock-in allele in combination with Rosa26-LacZ and Rosa26-tdTomato reporter strains showed that Lgr5-expressing cells gave rise to taste cells, perigemmal cells, along with self-renewing cells at the bottom of trench areas at the base of circumvallate and foliate papillae. Moreover, using subtype-specific taste markers, we found that Lgr5-expressing cell progeny include all three major types of adult taste cells. Our results indicate that Lgr5 may mark adult taste stem or progenitor cells in the posterior portion of the tongue. PMID:23377989
Lgr5-EGFP marks taste bud stem/progenitor cells in posterior tongue.
Yee, Karen K; Li, Yan; Redding, Kevin M; Iwatsuki, Ken; Margolskee, Robert F; Jiang, Peihua
2013-05-01
Until recently, reliable markers for adult stem cells have been lacking for many regenerative mammalian tissues. Lgr5 (leucine-rich repeat-containing G-protein-coupled receptor 5) has been identified as a marker for adult stem cells in intestine, stomach, and hair follicle; Lgr5-expressing cells give rise to all types of cells in these tissues. Taste epithelium also regenerates constantly, yet the identity of adult taste stem cells remains elusive. In this study, we found that Lgr5 is strongly expressed in cells at the bottom of trench areas at the base of circumvallate (CV) and foliate taste papillae and weakly expressed in the basal area of taste buds and that Lgr5-expressing cells in posterior tongue are a subset of K14-positive epithelial cells. Lineage-tracing experiments using an inducible Cre knockin allele in combination with Rosa26-LacZ and Rosa26-tdTomato reporter strains showed that Lgr5-expressing cells gave rise to taste cells, perigemmal cells, along with self-renewing cells at the bottom of trench areas at the base of CV and foliate papillae. Moreover, using subtype-specific taste markers, we found that Lgr5-expressing cell progeny include all three major types of adult taste cells. Our results indicate that Lgr5 may mark adult taste stem or progenitor cells in the posterior portion of the tongue. Copyright © 2013 AlphaMed Press.
Classification of conductance traces with recurrent neural networks
NASA Astrophysics Data System (ADS)
Lauritzen, Kasper P.; Magyarkuti, András; Balogh, Zoltán; Halbritter, András; Solomon, Gemma C.
2018-02-01
We present a new automated method for structural classification of the traces obtained in break junction experiments. Using recurrent neural networks trained on the traces of minimal cross-sectional area in molecular dynamics simulations, we successfully separate the traces into two classes: point contact or nanowire. This is done without any assumptions about the expected features of each class. The trained neural network is applied to experimental break junction conductance traces, and it separates the classes as well as the previously used experimental methods. The effect of using partial conductance traces is explored, and we show that the method performs equally well using full or partial traces (as long as the trace just prior to breaking is included). When only the initial part of the trace is included, the results are still better than random chance. Finally, we show that the neural network classification method can be used to classify experimental conductance traces without using simulated results for training, but instead training the network on a few representative experimental traces. This offers a tool to recognize some characteristic motifs of the traces, which can be hard to find by simple data selection algorithms.
Reflectance model of a plant leaf
NASA Technical Reports Server (NTRS)
Kumar, R.; Silva, L.
1973-01-01
A light ray, incident at 5 deg to the normal, is geometrically plotted through the drawing of the cross section of a soybean leaf using Fresnel's Equations and Snell's Law. The optical mediums of the leaf considered for ray tracing are: air, cell sap, chloroplast, and cell wall. The above ray is also drawn through the same leaf cross section considering cell wall and air as the only optical mediums. The values of the reflection and transmission found from ray tracing agree closely with the experimental results obtained using a Beckman DK-2A Spectroreflectometer. Similarly a light ray, incident at about 60 deg to the normal, is drawn through the palisade cells of a soybean leaf to illustrate the pathway of light, incident at an oblique angle, through the palisade cells.
Lineage tracing of dlx1a/2a and dlx5a/6a expressing cells in the developing zebrafish brain.
Solek, Cynthia M; Feng, Shengrui; Perin, Sofia; Weinschutz Mendes, Hellen; Ekker, Marc
2017-07-01
Lineage tracing of specific populations of progenitor cells provides crucial information about developmental programs. Four members of the Dlx homeobox gene family, Dlx1,2, 5 and 6, are involved in the specification of γ-aminobutyric acid (GABA)ergic neurons in the vertebrate forebrain. Orthologous genes in mammals and teleost show similarities in expression patterns and transcriptional regulation mechanisms. We have used lineage tracing to permanently label dlx-expressing cells in the zebrafish and have characterized the progeny of these cells in the larva and in the juvenile and adult brain. We have found that dlx1a/2a and dlx5a/6a expressing progenitors give rise, for the most part, to small populations of cells which constitute only a small proportion of GABAergic cells in the adult brain tissue. Moreover, some of the cells do not acquire a neuronal phenotype suggesting that, regardless of the time a cell expresses dlx genes in the brain, it can potentially give rise to cells other than neurons. In some instances, labeling larval dlx5a/6a-expressing cells, but not dlx1a/2a-expressing cells, results in massively expanding, widespread clonal expansion throughout the adult brain. Our data provide a detailed lineage analysis of the dlx1a/2a and dlx5a/6a expressing progenitors in the zebrafish brain and lays the foundation for further characterization of the role of these transcription factors beyond the specification of GABAergic neurons. Copyright © 2017 Elsevier Inc. All rights reserved.
Pacelli, Claudia; Selbmann, Laura; Zucconi, Laura; De Vera, Jean-Pierre; Rabbow, Elke; Horneck, Gerda; de la Torre, Rosa; Onofri, Silvano
2017-06-01
The search for traces of extinct or extant life in extraterrestrial environments is one of the main goals for astrobiologists; due to their ability to withstand stress producing conditions, extremophiles are perfect candidates for astrobiological studies. The BIOMEX project aims to test the ability of biomolecules and cell components to preserve their stability under space and Mars-like conditions, while at the same time investigating the survival capability of microorganisms. The experiment has been launched into space and is being exposed on the EXPOSE-R2 payload, outside of the International Space Station (ISS) over a time-span of 1.5 years. Along with a number of other extremophilic microorganisms, the Antarctic cryptoendolithic black fungus Cryomyces antarcticus CCFEE 515 has been included in the experiment. Before launch, dried colonies grown on Lunar and Martian regolith analogues were exposed to vacuum, irradiation and temperature cycles in ground based experiments (EVT1 and EVT2). Cultural and molecular tests revealed that the fungus survived on rock analogues under space and simulated Martian conditions, showing only slight ultra-structural and molecular damage.
NASA Astrophysics Data System (ADS)
Pacelli, Claudia; Selbmann, Laura; Zucconi, Laura; De Vera, Jean-Pierre; Rabbow, Elke; Horneck, Gerda; de la Torre, Rosa; Onofri, Silvano
2017-06-01
The search for traces of extinct or extant life in extraterrestrial environments is one of the main goals for astrobiologists; due to their ability to withstand stress producing conditions, extremophiles are perfect candidates for astrobiological studies. The BIOMEX project aims to test the ability of biomolecules and cell components to preserve their stability under space and Mars-like conditions, while at the same time investigating the survival capability of microorganisms. The experiment has been launched into space and is being exposed on the EXPOSE-R2 payload, outside of the International Space Station (ISS) over a time-span of 1.5 years. Along with a number of other extremophilic microorganisms, the Antarctic cryptoendolithic black fungus Cryomyces antarcticus CCFEE 515 has been included in the experiment. Before launch, dried colonies grown on Lunar and Martian regolith analogues were exposed to vacuum, irradiation and temperature cycles in ground based experiments (EVT1 and EVT2). Cultural and molecular tests revealed that the fungus survived on rock analogues under space and simulated Martian conditions, showing only slight ultra-structural and molecular damage.
Automated Tracing of Horizontal Neuron Processes During Retinal Development
DOE Office of Scientific and Technical Information (OSTI.GOV)
Kerekes, Ryan A; Martins, Rodrigo; Dyer, Michael A
2011-01-01
In the developing mammalian retina, horizontal neurons undergo a dramatic reorganization oftheir processes shortly after they migrate to their appropriate laminar position. This is an importantprocess because it is now understood that the apical processes are important for establishing theregular mosaic of horizontal cells in the retina and proper reorganization during lamination isrequired for synaptogenesis with photoreceptors and bipolar neurons. However, this process isdifficult to study because the analysis of horizontal neuron anatomy is labor intensive and time-consuming. In this paper, we present a computational method for automatically tracing the three-dimensional (3-D) dendritic structure of horizontal retinal neurons in two-photonmore » laser scanningmicroscope (TPLSM) imagery. Our method is based on 3-D skeletonization and is thus able topreserve the complex structure of the dendritic arbor of these cells. We demonstrate theeffectiveness of our approach by comparing our tracing results against two sets of semi-automatedtraces over a set of 10 horizontal neurons ranging in age from P1 to P5. We observe an averageagreement level of 81% between our automated trace and the manual traces. This automatedmethod will serve as an important starting point for further refinement and optimization.« less
Cell Phones ≠ Self and Other Problems with Big Data Detection and Containment during Epidemics.
Erikson, Susan L
2018-03-08
Evidence from Sierra Leone reveals the significant limitations of big data in disease detection and containment efforts. Early in the 2014-2016 Ebola epidemic in West Africa, media heralded HealthMap's ability to detect the outbreak from newsfeeds. Later, big data-specifically, call detail record data collected from millions of cell phones-was hyped as useful for stopping the disease by tracking contagious people. It did not work. In this article, I trace the causes of big data's containment failures. During epidemics, big data experiments can have opportunity costs: namely, forestalling urgent response. Finally, what counts as data during epidemics must include that coming from anthropological technologies because they are so useful for detection and containment. © 2018 The Authors Medical Anthropology Quarterly published by Wiley Periodicals, Inc. on behalf of American Anthropological Association.
Subseafloor Microbial Life in Venting Fluids from the Mid Cayman Rise Hydrothermal System
NASA Astrophysics Data System (ADS)
Huber, J. A.; Reveillaud, J.; Reddington, E.; McDermott, J. M.; Sylva, S. P.; Breier, J. A.; German, C. R.; Seewald, J.
2012-12-01
In hard rock seafloor environments, fluids emanating from hydrothermal vents are one of the best windows into the subseafloor and its resident microbial community. The functional consequences of an extensive population of microbes living in the subseafloor remains unknown, as does our understanding of how these organisms interact with one another and influence the biogeochemistry of the oceans. Here we report the abundance, activity, and diversity of microbes in venting fluids collected from two newly discovered deep-sea hydrothermal vents along the ultra-slow spreading Mid-Cayman Rise (MCR). Fluids for geochemical and microbial analysis were collected from the Von Damm and Piccard vent fields, which are located within 20 km of one another, yet have extremely different thermal, geological, and depth regimes. Geochemical data indicates that both fields are highly enriched in volatiles, in particular hydrogen and methane, important energy sources for and by-products of microbial metabolism. At both sites, total microbial cell counts in the fluids ranged in concentration from 5 x 10 4 to 3 x 10 5 cells ml-1 , with background seawater concentrations of 1-2 x 10 4 cells ml-1 . In addition, distinct cell morphologies and clusters of cells not visible in background seawater were seen, including large filaments and mineral particles colonized by microbial cells. These results indicate local enrichments of microbial communities in the venting fluids, distinct from background populations, and are consistent with previous enumerations of microbial cells in venting fluids. Stable isotope tracing experiments were used to detect utilization of acetate, formate, and dissolve inorganic carbon and generation of methane at 70 °C under anaerobic conditions. At Von Damm, a putatively ultra-mafic hosted site located at ~2200 m with a maximum temperature of 226 °C, stable isotope tracing experiments indicate methanogenesis is occurring in most fluid samples. No activity was detected in Piccard vent fluids, a basalt-hosted black smoker site located at ~4950 m with a maximum temperature of 403 °C. However, hyperthermophilic and thermophilic heterotrophs of the genus Thermococcus were isolated from Piccard vent fluids, but not Von Damm. These obligate anaerobes, growing optimally at 55-90 °C, are ubiquitous at hydrothermal systems and serve as a readily cultivable indicator organism of subseafloor populations. Finally, molecular analysis of vent fluids is on-going and will define the microbial population structure in this novel ecosystem and allow for direct comparisons with other deep-sea and subsurface habitats as part of our continuing efforts to explore the deep microbial biosphere on Earth.
DOE Office of Scientific and Technical Information (OSTI.GOV)
Pearce, R.J.H.; Bell, A.C.; Brennan, D.
'Trace Tritium Experiments' (TTE) were successfully performed on JET in 2003. The Campaign marked the first use of tritium in JET plasmas since the Deuterium-Tritium Experiment (DTE1) Campaign in 1997, and was the first use of tritium in experiments under the EFDA organisation with the UKAEA as JET Operator. The safety and regulatory preparations for the experiment were extensive. Since JET has been operated by the UKAEA the operations have followed the model of a licensed nuclear site. The safe operation of the JET torus is demonstrated in a safety case. Key Safety Management Requirement (KSMR) and Key Safety Relatedmore » Equipment (KSRE) are identified in the Safety Case for DT operation. The safe operation of the torus is within the bounds of, and under the control of, an Authority to Operate (ATO). New technical challenges were presented by the need to inject and account for small quantities of tritium in very short pulses ({approx}80ms), with an accurate time stamp. The safety and operational management of the campaign are described. Valuable lessons were learned which would help in running future experiments. It is concluded that JET is in a strong position to run future trace tritium and full DT discharges.« less
Introductory Laboratory Exercises in Radiobiology
ERIC Educational Resources Information Center
Williams, J. R. Parry; Servant, D. M.
1970-01-01
Describes experiments suitable for introducing use of radioisotopes in biology. Includes demonstrations of tracing food chains, uptake of ions by plants, concentration of elements by insects, tracing photosynthetic reactions, activation analysis of copper, and somatic and genetic effects. Uses autoradiographic and counting techniques. (AL)
Lee, Jin Hyung
2011-01-01
Despite the overwhelming need, there has been a relatively large gap in our ability to trace network level activity across the brain. The complex dense wiring of the brain makes it extremely challenging to understand cell-type specific activity and their communication beyond a few synapses. Recent development of the optogenetic functional magnetic resonance imaging (ofMRI) provides a new impetus for the study of brain circuits by enabling causal tracing of activities arising from defined cell types and firing patterns across the whole brain. Brain circuit elements can be selectively triggered based on their genetic identity, cell body location, and/or their axonal projection target with temporal precision while the resulting network response is monitored non-invasively with unprecedented spatial and temporal accuracy. With further studies including technological innovations to bring ofMRI to its full potential, ofMRI is expected to play an important role in our system-level understanding of the brain circuit mechanism. PMID:22046160
Chapple, Richard H.; Tseng, Yu-Jung; Hu, Tianyuan; Kitano, Ayumi; Takeichi, Makiko; Hoegenauer, Kevin A.
2018-01-01
Characterization of hematopoietic stem cells (HSCs) has advanced largely owing to transplantation assays, in which the developmental potential of HSCs is assessed generally in nonhomeostatic conditions. These studies established that adult HSCs extensively contribute to multilineage hematopoietic regeneration upon transplantation. On the contrary, recent studies performing lineage tracing of HSCs under homeostatic conditions have shown that adult HSCs may contribute far less to steady-state hematopoiesis than would be anticipated based on transplantation assays. Here, we used 2 independent HSC-lineage–tracing models to examine the contribution of adult HSCs to steady-state hematopoiesis. We show that adult HSCs contribute robustly to steady-state hematopoiesis, exhibiting faster efflux toward the myeloid lineages compared with lymphoid lineages. Platelets were robustly labeled by HSCs, reaching the same level of labeling as HSCs by 1 year of chase. Our results support the view that adult HSCs contribute to the continuous influx of blood cells during steady-state hematopoiesis. PMID:29848758
Xiao, Yi; Tholen, Danny; Zhu, Xin-Guang
2016-11-01
Leaf photosynthesis is determined by biochemical properties and anatomical features. Here we developed a three-dimensional leaf model that can be used to evaluate the internal light environment of a leaf and its implications for whole-leaf electron transport rates (J). This model includes (i) the basic components of a leaf, such as the epidermis, palisade and spongy tissues, as well as the physical dimensions and arrangements of cell walls, vacuoles and chloroplasts; and (ii) an efficient forward ray-tracing algorithm, predicting the internal light environment for light of wavelengths between 400 and 2500nm. We studied the influence of leaf anatomy and ambient light on internal light conditions and J The results show that (i) different chloroplasts can experience drastically different light conditions, even when they are located at the same distance from the leaf surface; (ii) bundle sheath extensions, which are strips of parenchyma, collenchyma or sclerenchyma cells connecting the vascular bundles with the epidermis, can influence photosynthetic light-use efficiency of leaves; and (iii) chloroplast positioning can also influence the light-use efficiency of leaves. Mechanisms underlying leaf internal light heterogeneity and implications of the heterogeneity for photoprotection and for the convexity of the light response curves are discussed. © The Author 2016. Published by Oxford University Press on behalf of the Society for Experimental Biology.
Bacterial ethane formation from reduced, ethylated sulfur compounds in anoxic sediments
Oremland, R.S.; Whiticar, Michael J.; Strohmaier, F.E.; Kiene, R.P.
1988-01-01
Trace levels of ethane were produced biologically in anoxic sediment slurries from five chemically different aquatic environments. Gases from these locations displayed biogenic characteristics, having 12C-enriched values of ??13CH4 (-62 to -86%.), ??13C2H6 (-35 to -55%.) and high ratios (720 to 140,000) of CH4 [C2H6 + C3H8]. Endogenous production of ethane by slurries was inhibited by autoclaving or by addition of the inhibitor of methanogenic bacteria, 2-bromoethanesulfonic acid (BES). Ethane formation was stimulated markedly by ethanethiol (ESH), and, to a lesser extent, by diethylsulfide (DES). Formation of methane and ethane in ESH- or DES-amended slurries was blocked by BES. Experiments showed that ethionine (or an analogous compound) could be a precursor of ESH. Ethylamine or ethanol additions to slurries caused only a minor stimulation of ethane formation. Similarly, propanethiol additions resulted in only a minor enhancement of propane formation. Cell suspensions of a methyltrophic methanogen produced traces of ethane when incubated in the presence of DES, although the organism did not grow on this compound. These results indicate that methanogenic bacteria produce ethane from the traces of ethylated sulfur compounds present in recent sediments. Preliminary estimates of stable carbon isotope fractionation associated with sediment methane formation from dimethylsulfide was about 40%., while ethane formation from DES and ESH was only 4. 6 and 6.5%., respectively. ?? 1988.
Zhang, Hua; Zheng, Wenjing; Shen, Yan; Adhikari, Deepak; Ueno, Hiroo; Liu, Kui
2012-07-31
It has been generally accepted for more than half a century that, in most mammalian species, oocytes cannot renew themselves in postnatal or adult life, and that the number of oocytes is already fixed in fetal or neonatal ovaries. This assumption, however, has been challenged over the past decade. In this study, we have taken an endogenous genetic approach to this question and generated a multiple fluorescent Rosa26(rbw/+);Ddx4-Cre germline reporter mouse model for in vivo and in vitro tracing of the development of female germline cell lineage. Through live cell imaging and de novo folliculogenesis experiments, we show that the Ddx4-expressing cells from postnatal mouse ovaries did not enter mitosis, nor did they contribute to oocytes during de novo folliculogenesis. Our results provide evidence that supports the traditional view that no postnatal follicular renewal occurs in mammals, and no mitotically active Ddx4-expressing female germline progenitors exist in postnatal mouse ovaries.
NASA Astrophysics Data System (ADS)
Wang, Zheng-Fang; Zhang, Qing; Gong, Min
1995-12-01
Laboratory culture experiments showed that <100μ mol/L nitrate, amonium or mixture of amino acids promote the growth of the red tide organism Prorocentrum micans Ehrenb, but that >100μmol/L of ammonium, or mixture of glycine and glutamate was harmful to growth, and that orthophosphate was P. micans’ main phosphorous source in the ocean. Presence of 80μ mol/L EDTA, 0.5 to 1 μmol/L Fe3+, 1.0 to 20.0 μ mol/L Mn2+ 0.1 to 0.4 μmol/L Co2+ in the culture medium could improve the growth of P. micans. Vitamin B1 promoted growth, but vitamin B12 and biotin did not. The estimated minimum cell quotas ( q o) for nitrogen and phosphorus being 0.74 pmole/cell and 0.045 pmole/cell show that phosphorus (more than nitrogen) limits the growth of P. micans in the study area.
NASA Astrophysics Data System (ADS)
Loppini, A.; Pedersen, M. G.; Braun, M.; Filippi, S.
2017-09-01
The importance of gap-junction coupling between β cells in pancreatic islets is well established in mouse. Such ultrastructural connections synchronize cellular activity, confine biological heterogeneity, and enhance insulin pulsatility. Dysfunction of coupling has been associated with diabetes and altered β -cell function. However, the role of gap junctions between human β cells is still largely unexplored. By using patch-clamp recordings of β cells from human donors, we previously estimated electrical properties of these channels by mathematical modeling of pairs of human β cells. In this work we revise our estimate by modeling triplet configurations and larger heterogeneous clusters. We find that a coupling conductance in the range 0.005 -0.020 nS/pF can reproduce experiments in almost all the simulated arrangements. We finally explore the consequence of gap-junction coupling of this magnitude between β cells with mutant variants of the ATP-sensitive potassium channels involved in some metabolic disorders and diabetic conditions, translating studies performed on rodents to the human case. Our results are finally discussed from the perspective of therapeutic strategies. In summary, modeling of more realistic clusters with more than two β cells slightly lowers our previous estimate of gap-junction conductance and gives rise to patterns that more closely resemble experimental traces.
Cellular basis of gastrulation in the sand dollar Scaphechinus mirabilis.
Kominami, T; Takata, H
2000-12-01
The processes of gastrulation in the sand dollar Scaphechinus mirabilis are quite different from those in regular echinoids. In this study, we explored the cellular basis of gastrulation in this species with several methods. Cell-tracing experiments revealed that the prospective endodermal cells were convoluted throughout the invagination processes. Histological observation showed that the ectodermal layer remained thickened, and the vegetal cells retained an elongated shape until the last step of invagination. Further, most of the vegetal ectodermal cells were skewed or distorted. Wedge-shaped cells were common in the vegetal ectoderm, especially at the subequatorial region. In these embryos, unlike the embryos of regular echinoids, secondary mesenchyme cells did not seem to exert the force to pull up the archenteron toward the inner surface of the apical plate. In fact, the archenteron cells were not stretched along the axis of elongation and were in close contact with each other. Here we found that gastrulation was completely blocked when the embryos were attached to a glass dish coated with poly-L-lysine, in which the movement of the ectodermal layer was inhibited. These results suggest that a force generated by the thickened ectoderm, rather than rearrangement of the archenteron cells, may play a key role in the archenteron elongation in S. mirabilis embryos.
Zhou, Shukui; Yin, Ting; Zou, Qingsong; Zhang, Kaile; Gao, Guo; Shapter, Joseph G; Huang, Peng; Fu, Qiang
2017-02-21
Cell sheet therapy has emerged as a potential therapeutic option for reparation and reconstruction of damaged tissues and organs. However, an effective means to assess the fate and distribution of transplanted cell sheets in a serial and noninvasive manner is still lacking. To investigate the feasibility of tracking Adipose derived stem cells (ADSCs) sheet in vivo using ultrasmall super-paramagnetic Fe 3 O 4 nanoparticles (USPIO), canine ADSCs were cultured and incubated with USPIO and 0.75 μg/ml Poly-L-Lysine (PLL) for 12 h. Labeling efficiency, cell viability, apoptotic cell rate were assessed to screen the optimum concentrations of USPIO for best labeling ADSCs. The results showed ADSCs were labeled by USPIO at an iron dose of 50 μg/ml for a 12 h incubation time, which can most efficiently mark cells and did not impair the cell survival, self-renewal, and proliferation capacity. USPIO-labeled ADSCs sheets can be easily and clearly detected in vivo and have persisted for at least 12 weeks. Our experiment confirmed USPIO was feasible for in vivo labeling of the ADSCs sheets with the optimal concentration of 50 μg Fe/ml and the tracing time is no less than 12 weeks.
On modeling weak sinks in MODPATH
Abrams, Daniel B.; Haitjema, Henk; Kauffman, Leon J.
2012-01-01
Regional groundwater flow systems often contain both strong sinks and weak sinks. A strong sink extracts water from the entire aquifer depth, while a weak sink lets some water pass underneath or over the actual sink. The numerical groundwater flow model MODFLOW may allow a sink cell to act as a strong or weak sink, hence extracting all water that enters the cell or allowing some of that water to pass. A physical strong sink can be modeled by either a strong sink cell or a weak sink cell, with the latter generally occurring in low resolution models. Likewise, a physical weak sink may also be represented by either type of sink cell. The representation of weak sinks in the particle tracing code MODPATH is more equivocal than in MODFLOW. With the appropriate parameterization of MODPATH, particle traces and their associated travel times to weak sink streams can be modeled with adequate accuracy, even in single layer models. Weak sink well cells, on the other hand, require special measures as proposed in the literature to generate correct particle traces and individual travel times and hence capture zones. We found that the transit time distributions for well water generally do not require special measures provided aquifer properties are locally homogeneous and the well draws water from the entire aquifer depth, an important observation for determining the response of a well to non-point contaminant inputs.
NASA Astrophysics Data System (ADS)
Devès, Guillaume; Cohen-Bouhacina, Touria; Ortega, Richard
2004-10-01
We used the nuclear microprobe techniques, micro-PIXE (particle-induced X-ray emission), micro-RBS (Rutherford backscattering spectrometry) and scanning transmission ion microscopy (STIM) in order to perform the characterization of trace element content and spatial distribution within biological samples (dehydrated cultured cells, tissues). The normalization of PIXE results was usually expressed in terms of sample dry mass as determined by micro-RBS recorded simultaneously to micro-PIXE. However, the main limit of RBS mass measurement is the sample mass loss occurring during irradiation and which could be up to 30% of the initial sample mass. We present here a new methodology for PIXE normalization and quantitative analysis of trace element within biological samples based on dry mass measurement performed by mean of STIM. The validation of STIM cell mass measurements was obtained in comparison with AFM sample thickness measurements. Results indicated the reliability of STIM mass measurement performed on biological samples and suggested that STIM should be performed for PIXE normalization. Further information deriving from direct confrontation of AFM and STIM analysis could as well be obtained, like in situ measurements of cell specific gravity within cells compartment (nucleolus and cytoplasm).
Hoo, Henny; Hashidoko, Yasuyuki; Islam, Md. Tofazzal; Tahara, Satoshi
2004-01-01
Mg2+ is one of the essential elements for bacterial cell growth. The presence of the magnesium cation (Mg2+) in various concentrations often affects cell growth restoration in plant-associating bacteria. This study attempted to determine whether Mg2+ levels in Sphingomonas yanoikuyae EC-S001 affected cell growth restoration in the host plant and what the threshold level is. S. yanoikuyae EC-S001, isolated from the rhizoplane of spinach seedlings grown from surface-sterilized seeds under aseptic conditions, displayed uniform dispersion and attachment throughout the rhizoplane and phylloplane of the host seedlings. S. yanoikuyae EC-S001 did not grow in potato-dextrose broth medium but grew well in an aqueous extract of spinach leaves. Chemical investigation of the growth factor in the spinach leaf extract led to identification of the active principle as the magnesium cation. A concentration of ca. 0.10 mM Mg2+ or more allowed S. yanoikuyae EC-S001 to grow in potato-dextrose broth medium. Some saprophytic and/or diazotrophic bacteria used in our experiment were found to have diverse threshold levels for their Mg2+ requirements. For example, Burkholderia cepacia EC-K014, originally isolated from the rhizoplane of a Melastoma sp., could grow even in Mg2+-free Hoagland's no. 2 medium with saccharose and glutamine (HSG medium) and requires a trace level of Mg2+ for its growth. In contrast, S. yanoikuyae EC-S001, together with Bacillus subtilis IFO12113, showed the most drastic restoring responses to subsequent addition of 0.98 mM Mg2+ to Mg2+-free HSG medium. Our studies concluded that Mg2+ is more than just the essential trace element needed for cell growth restoration in S. yanoikuyae EC-S001 and that certain nonculturable bacteria may require a higher concentration of Mg2+ or another specific essential element for their growth. PMID:15345402
Trace detection of specific viable bacteria using tetracysteine-tagged bacteriophages.
Wu, Lina; Luan, Tian; Yang, Xiaoting; Wang, Shuo; Zheng, Yan; Huang, Tianxun; Zhu, Shaobin; Yan, Xiaomei
2014-01-07
Advanced methods are urgently needed to determine the identity and viability of trace amounts of pathogenic bacteria in a short time. Existing approaches either fall short in the accurate assessment of microbial viability or lack specificity in bacterial identification. Bacteriophages (or phages for short) are viruses that exclusively infect bacterial host cells with high specificity. As phages infect and replicate only in living bacterial hosts, here we exploit the strategy of using tetracysteine (TC)-tagged phage in combination with biarsenical dye to the discriminative detection of viable target bacteria from dead target cells and other viable but nontarget bacterial cells. Using recombinant M13KE-TC phage and Escherichia coli ER2738 as a model system, distinct differentiation between individual viable target cells from dead target cells was demonstrated by flow cytometry and fluorescence microscopy. As few as 1% viable E. coli ER2738 can be accurately quantified in a mix with dead E. coli ER2738 by flow cytometry. With fluorescence microscopic measurement, specific detection of as rare as 1 cfu/mL original viable target bacteria was achieved in the presence of a large excess of dead target cells and other viable but nontarget bacterial cells in 40 mL artificially contaminated drinking water sample in less than 3 h. This TC-phage-FlAsH approach is sensitive, specific, rapid, and simple, and thus shows great potential in water safety monitoring, health surveillance, and clinical diagnosis of which trace detection and identification of viable bacterial pathogens is highly demanded.
Predictions of Critical Heat Flux in Annular Pipes with TRACEv4.160 code
DOE Office of Scientific and Technical Information (OSTI.GOV)
Jasiulevicius, Audrius; Macian-Juan, Rafael
2006-07-01
This paper presents the assessment of TRACE (version v4.160) against the Critical Heat Flux (CHF) experiments in annular tubes performed at the Royal Institute of Technology (KTH) in Stockholm, Sweden. The experimental database includes data for coolant mass fluxes between 250 and 2500 kg/m{sup 2}s and inlet subcooling of 10 and 40 K at a pressure of 70 bar. The work presented in this paper supplements the calculations of single round tube experiments carried out earlier and provides a broader scope of validated geometries. In addition to the Biasi and CISE-GE CHF correlations available in the code, a number ofmore » experimental points at low flow conditions are available for the annular geometry experiments, which also permitted the assessment of the Biasi/Zuber CHF correlation used in TRACE v4.160 for low flow conditions. Experiments with different axial power distribution were simulated and the effects of the axial power profile and the coolant inlet subcooling on the TRACE predictions were investigated. The results of this work show that the Biasi/Zuber correlation provides good estimation of the CHF at 70 bar, and, for the same conditions, the simulation of the annular experiments resulted in the calculation of lower CHF values compared to single-tube experiments. The analysis of the performance of the standard TRACE CHF correlations shows that the CISE-GE correlation yields critical qualities (quality at CHF) closer to the experimental values at 70 bar than the Biasi correlation for annular flow conditions. Regarding the power profile, the results of the TRACE calculations seem to be very sensitive to its shape, since, depending on the profile, different accuracies in the predictions were noted while other system conditions remained constant. The inlet coolant subcooling was also an important factor in the accuracy of TRACE CHF predictions. Thus, an increase in the inlet subcooling led to a clear improvement in the estimation of the critical quality with both Biasi and CISE-GE correlations. To complement the work, three additional CHF correlations were implemented in TRACE v4.160, namely the Bowring, Tong W-3 and Levitan-Lantsman CHF models, in order to assess the applicability of these correlations to simulate the CHF in annular tubes. The improvement of CHF predictions for low coolant mass flows (up to 1500 kg/m{sup 2}s) is noted when applying Bowring CHF correlation. However, the increase in the inlet subcooling increases the error in predicted critical quality with the Bowring correlation. The Levitan-Lantsman and Tong-W-3 correlations provide results similar to the Biasi model. Therefore, the most correct CHF predictions among the investigated correlations were obtained using CISE-GE model in the standard TRAC v4.160 code. (authors)« less
Trace Elements in Parenteral Nutrition: Considerations for the Prescribing Clinician
Jin, Jennifer; Mulesa, Leanne; Carrilero Rouillet, Mariana
2017-01-01
Trace elements (TEs) are an essential component of parenteral nutrition (PN). Over the last few decades, there has been increased experience with PN, and with this knowledge more information about the management of trace elements has become available. There is increasing awareness of the effects of deficiencies and toxicities of certain trace elements. Despite this heightened awareness, much is still unknown in terms of trace element monitoring, the accuracy of different assays, and current TE contamination of solutions. The supplementation of TEs is a complex and important part of the PN prescription. Understanding the role of different disease states and the need for reduced or increased doses is essential. Given the heterogeneity of the PN patients, supplementation should be individualized. PMID:28452962
Trace Elements in Parenteral Nutrition: Considerations for the Prescribing Clinician.
Jin, Jennifer; Mulesa, Leanne; Carrilero Rouillet, Mariana
2017-04-28
Trace elements (TEs) are an essential component of parenteral nutrition (PN). Over the last few decades, there has been increased experience with PN, and with this knowledge more information about the management of trace elements has become available. There is increasing awareness of the effects of deficiencies and toxicities of certain trace elements. Despite this heightened awareness, much is still unknown in terms of trace element monitoring, the accuracy of different assays, and current TE contamination of solutions. The supplementation of TEs is a complex and important part of the PN prescription. Understanding the role of different disease states and the need for reduced or increased doses is essential. Given the heterogeneity of the PN patients, supplementation should be individualized.
Quantitative assessment of fibroblast growth factor receptor 1 expression in neurons and glia.
Choubey, Lisha; Collette, Jantzen C; Smith, Karen Müller
2017-01-01
Fibroblast growth factors (FGFs) and their receptors (FGFRs) have numerous functions in the developing and adult central nervous system (CNS). For example, the FGFR1 receptor is important for proliferation and fate specification of radial glial cells in the cortex and hippocampus, oligodendrocyte proliferation and regeneration, midline glia morphology and soma translocation, Bergmann glia morphology, and cerebellar morphogenesis. In addition, FGFR1 signaling in astrocytes is required for postnatal maturation of interneurons expressing parvalbumin (PV). FGFR1 is implicated in synapse formation in the hippocampus, and alterations in the expression of Fgfr1 and its ligand, Fgf2 accompany major depression. Understanding which cell types express Fgfr1 during development may elucidate its roles in normal development of the brain as well as illuminate possible causes of certain neuropsychiatric disorders. Here, we used a BAC transgenic reporter line to trace Fgfr1 expression in the developing postnatal murine CNS. The specific transgenic line employed was created by the GENSAT project, tgFGFR1-EGFPGP338Gsat , and includes a gene encoding enhanced green fluorescent protein ( EGFP ) under the regulation of the Fgfr1 promoter, to trace Fgfr1 expression in the developing CNS. Unbiased stereological counts were performed for several cell types in the cortex and hippocampus. This model reveals that Fgfr1 is primarily expressed in glial cells, in both astrocytes and oligodendrocytes, along with some neurons. Dual labeling experiments indicate that the proportion of GFP+ ( Fgfr1 +) cells that are also GFAP+ increases from postnatal day 7 (P7) to 1 month, illuminating dynamic changes in Fgfr1 expression during postnatal development of the cortex. In postnatal neurogenic areas, GFP expression was also observed in SOX2, doublecortin (DCX), and brain lipid-binding protein (BLBP) expressing cells. Fgfr1 is also highly expressed in DCX positive cells of the dentate gyrus (DG), but not in the rostral migratory stream. Fgfr1 driven GFP was also observed in tanycytes and GFAP+ cells of the hypothalamus, as well as in Bergmann glia and astrocytes of the cerebellum. The tgFGFR1-EGFPGP338Gsat mouse model expresses GFP that is congruent with known functions of FGFR1, including hippocampal development, glial cell development, and stem cell proliferation. Understanding which cell types express Fgfr1 may elucidate its role in neuropsychiatric disorders and brain development.
Effects of an injectable trace mineral supplement on first-service conception rate of dairy cows.
Vanegas, J A; Reynolds, J; Atwill, E R
2004-11-01
A total of 825 dairy cows from a commercial dairy farm in central California were used to evaluate effects of one or 2 doses of an injectable trace mineral supplement containing 20 mg/mL of zinc, 20 mg/mL of manganese, 5 mg/mL of selenium, and 10 mg/mL of copper on first-service conception rate. Cows were randomly allocated into treatment or control group to either a single dose (experiment 1) or a double dose (experiment 2) of injected supplement. Allocation was based on days in lactation for experiment 1 and the length of gestation periods for experiment 2. In experiment 1, cows 38 to 45 d in lactation (n = 190) received a single dose of 5 mL of injected supplement. Similar cows were used as controls (n = 227). In experiment 2, cows and pregnant heifers received an initial injection of 5 mL of the mineral supplement from 2 to 3 wk precalving (n = 186). An equal dose was repeated 38 to 45 d in lactation. A similar group of cows and pregnant heifers served as controls for experiment 2 (n = 222). Health and reproductive events postcalving were recorded. In experiment 1, the odds of first-service conception were not significantly different for cows receiving a one-dose regimen of minerals compared with untreated control cows; conception rates were 26.8 and 27.5% for experiment 1 treatment and control groups, respectively. In experiment 1, the odds of first-service conception were significantly lower (odds ratio = 0.66) for cows and heifers given the 2-dose regimen of minerals compared with untreated controls; overall conception rates were 21.5 and 31.5% for experiment 2 treatment and control groups, respectively. In this intensively managed dairy herd, a single dose of injected trace minerals before breeding had no beneficial effects on first-service conception rate. However, dairy cows receiving a dose of trace minerals before calving and another dose before breeding had lower conception at first service.
The influence of carbon, sulfur, and silicon on trace element partitioning in iron alloys
NASA Astrophysics Data System (ADS)
Han, J.; Van Orman, J. A.; Crispin, K. L.; Ash, R. D.
2014-12-01
Non-metallic light elements are important constituents of planetary cores and have a strong influence on the partitioning behavior of trace elements. Planetary cores may contain a wide range of non-metallic light elements, including H, N, S, P, Si, and C. Under highly reducing conditions, such as those that are thought to have pertained during the formation of Mercury's core, Si and C, in addition to sulfur, may be particularly important constituents. Each of these elements may strongly effect and have a different impact on the partitioning behavior of trace elements but their combined effects on trace element partitioning have not been quantified. We investigated the partitioning behavior of more than 25 siderophile trace elements within the Fe-S-C-Si system with varying concentrations of C, S, and Si. The experiments were performed under pressures varying from 1 atm to 2 GPa and temperatures ranging from 1200˚C to 1450˚C. All experiments produced immiscible liquids, one enriched in Si and C, and the other predominantly FeS. We found some highly siderophile elements including Os, Ru, Ir, and Re are much more enriched in Fe-Si-C phase than in Fe-S phase, whereas other trace elements like V, Co, Ag, Hf, and Pb are enriched in S-rich phase. However, not all the trace elements enriched in Fe-Si-C phase are repelled by sulfur. Elements like Re and Ru could have different partitioning trends if sulfur concentration in S-rich phase rises. The partitioning behavior of these trace elements could enhance our understanding of the differentiation of Mercury's core under oxygen-poor conditions.
2017-01-01
As water scarcity intensifies, point-of-use and point-of-entry treatment may provide a means of exploiting locally available water resources that are currently considered to be unsafe for human consumption. Among the different classes of drinking water contaminants, toxic trace elements (e.g., arsenic and lead) pose substantial operational challenges for distributed drinking water treatment systems. Removal of toxic trace elements via adsorption onto iron oxides is an inexpensive and robust treatment method; however, the presence of metal-complexing ligands associated with natural organic matter (NOM) often prevents the formation of iron precipitates at the relatively low concentrations of dissolved iron typically present in natural water sources, thereby requiring the addition of iron which complicates the treatment process and results in a need to dispose of relatively large amounts of accumulated solids. A point-of-use treatment device consisting of a cathodic cell that produced hydrogen peroxide (H2O2) followed by an ultraviolet (UV) irradiation chamber was used to decrease colloid stabilization and metal-complexing capacity of NOM present in groundwater. Exposure to UV light altered NOM, converting ∼6 μM of iron oxides into settable forms that removed between 0.5 and 1 μM of arsenic (As), lead (Pb), and copper (Cu) from solution via adsorption. After treatment, changes in NOM consistent with the loss of iron-complexing carboxylate ligands were observed, including decreases in UV absorbance and shifts in the molecular composition of NOM to higher H/C and lower O/C ratios. Chronoamperometric experiments conducted in synthetic groundwater revealed that the presence of Ca2+ and Mg2+ inhibited intramolecular charge-transfer within photoexcited NOM, leading to substantially increased removal of iron and trace elements. PMID:29240414
Barazesh, James M; Prasse, Carsten; Wenk, Jannis; Berg, Stephanie; Remucal, Christina K; Sedlak, David L
2018-01-02
As water scarcity intensifies, point-of-use and point-of-entry treatment may provide a means of exploiting locally available water resources that are currently considered to be unsafe for human consumption. Among the different classes of drinking water contaminants, toxic trace elements (e.g., arsenic and lead) pose substantial operational challenges for distributed drinking water treatment systems. Removal of toxic trace elements via adsorption onto iron oxides is an inexpensive and robust treatment method; however, the presence of metal-complexing ligands associated with natural organic matter (NOM) often prevents the formation of iron precipitates at the relatively low concentrations of dissolved iron typically present in natural water sources, thereby requiring the addition of iron which complicates the treatment process and results in a need to dispose of relatively large amounts of accumulated solids. A point-of-use treatment device consisting of a cathodic cell that produced hydrogen peroxide (H 2 O 2 ) followed by an ultraviolet (UV) irradiation chamber was used to decrease colloid stabilization and metal-complexing capacity of NOM present in groundwater. Exposure to UV light altered NOM, converting ∼6 μM of iron oxides into settable forms that removed between 0.5 and 1 μM of arsenic (As), lead (Pb), and copper (Cu) from solution via adsorption. After treatment, changes in NOM consistent with the loss of iron-complexing carboxylate ligands were observed, including decreases in UV absorbance and shifts in the molecular composition of NOM to higher H/C and lower O/C ratios. Chronoamperometric experiments conducted in synthetic groundwater revealed that the presence of Ca 2+ and Mg 2+ inhibited intramolecular charge-transfer within photoexcited NOM, leading to substantially increased removal of iron and trace elements.
Presynaptic Inputs to Any CNS Projection Neuron Identified by Dual Recombinant Virus Infection
Bráz, João M.; Wang, Fan; Basbaum, Allan I.
2015-01-01
Although neuroanatomical tracing studies have defined the origin and targets of major projection neurons (PN) of the central nervous system (CNS), there is much less information about the circuits that influence these neurons. Recently, genetic approaches that use Cre recombinase-dependent viral vectors have greatly facilitated such circuit analysis, but these tracing approaches are limited by the availability of Cre-expressing mouse lines and the difficulty in restricting Cre expression to discrete regions of the CNS. Here, we illustrate an alternative approach to drive Cre expression specifically in defined subsets of CNS projection neurons, so as to map both direct and indirect presynaptic inputs to these cells. The method involves a combination of Cre-dependent transneuronal viral tracers that can be used in the adult and that does not require genetically modified mice. To trigger Cre-expression we inject a Cre-expressing adenovirus that is retrogradely transported to the projection neurons of interest. The region containing the retrogradely labeled projection neurons is next injected with Cre-dependent pseudorabies or rabies vectors, which results in labeling of poly- and monosynaptic neuronal inputs, respectively. In proof-of-concept experiments, we used this novel tracing system to study the circuits that engage projection neurons of the superficial dorsal horn of the spinal cord and trigeminal nucleus caudalis, neurons of the parabrachial nucleus of the dorsolateral pons that project to the amygdala and cortically-projecting neurons of the lateral geniculate nucleus. Importantly, because this dual viral tracing method does not require genetically derived Cre-expressing mouse lines, inputs to almost any projection system can be studied and the analysis can be performed in larger animals, such as the rat. PMID:26470056
Image Tracing: An Analysis of Its Effectiveness in Children's Pictorial Discrimination Learning
ERIC Educational Resources Information Center
Levin, Joel R.; And Others
1977-01-01
A total of 45 fifth grade students were the subjects of an experiment offering support for a component of learning strategy (memory imagery). Various theoretical explanations of the image-tracing phenomenon are considered, including depth of processing, dual coding and frequency. (MS)
Humans experience chronic cumulative trace-level exposure to mixtures of volatile, semi-volatile, and non-volatile polycyclic aromatic hydrocarbons (PAHs) present in the environment as by-products of combustion processes. Certain PAHs are known or suspected human carcinogens and ...
Development of high-efficiency solar cells on silicon web
NASA Technical Reports Server (NTRS)
Rohatgi, A.; Meier, D. L.; Campbell, R. B.; Seidensticker, R. G.; Rai-Choudhury, P.
1984-01-01
The development of high efficiency solar cells on a silicon web is discussed. Heat treatment effects on web quality; the influence of twin plane lamellae, trace impurities and stress on minority carrier lifetime; and the fabrication of cells are discussed.
Trace elements in children suffering from sickle cell anemia: A case-control study.
Sungu, Joseph K; Mukuku, Olivier; Mutombo, Augustin Mulangu; Mawaw, Paul; Aloni, Michel N; Luboya, Oscar N
2018-01-01
Information on serum albumin and trace elements among children suffering from Sickle Cell Anemia (SCA) was poorly documented in Africa. The aim of this study was to describe and to compare different values of trace elements with published reports from other parts of the world. We carried out a case-control study. Seventy-six steady state children suffering from SCA (Hb-SS). One hundred and fifty-two children were recruited with 76 (cases, Hb-SS) and 76 (control, Hb-AA) to compare the data. The mean age was 10.0 years (SD=5.4) in SCA children and 9.2 years (SD=4.7) in the control group. The mean level of zinc and magnesium were slightly lower in the Hb-SS group than in the Hb-AA group (P<.001). The first literature about trace elements in SCA is briefly reported in Central Africa. In our midst, zinc, magnesium, and selenium deficiencies underline the need for their systematic among all children with SCA to identify patients with these deficiencies and provide early management. © 2017 Wiley Periodicals, Inc.
Hormesis effect of trace metals on cultured normal and immortal human mammary cells.
Schmidt, Craig M; Cheng, Chun N; Marino, Angelo; Konsoula, Roula; Barile, Frank A
2004-06-01
An in vitro study was conducted to determine the effects of variable concentrations of trace metals on human cultured mammary cells. Monolayers of human mortal (MCF-12A) and immortal (MDA-MB231) mammary epithelial cells were incubated in the absence or presence of increasing concentrations of arsenic (As), mercury (Hg) and copper (Cu) for 24-h, 72-h, 4-d, and 7-d. The MTT assay was used to assess viability for all time periods and cell proliferation was monitored for 4-d and 7-d studies. Monolayers were also labeled with rhodamine-110 (R-6501), Sytox green, and Celltiter blue fluorescent dyes as indicators for intracellular esterase activity, nucleic acid staining, and cell reduction/viability, respectively. Total incubation time with chemical plus dyes was 24 h. For 24-h and 72-h studies, cells were seeded in 96-well plates, after which confluent monolayers were exposed to increasing concentrations of chemicals. For 4-d and 7-d studies, cells were seeded in 12-well plates at 1/3 confluent density (day 0) and exposed to increasing concentrations of metals on day 1. All cells were counted on days 4 and 7. In addition, test medium was removed from select groups of cultures on day 4, replaced with fresh medium in the absence of chemical (recovery studies), and assays were performed on day 7 as above. The data suggest that there is a consistent protective and/or stimulating effect of metals at the lowest concentrations in MCF-12A cells that is not observed in immortal MDA-MB231 cells. In fact, cell viability of MCF-12A cells is stimulated by otherwise equivalent inhibitory concentrations of As, Cu, and Hg on MDA-MB231 cells at 24-h. Whereas As and Hg suppress proliferation and viability in both cell lines after 4-d and 7-d of exposure, Cu enhances cell proliferation and viability of MCF-12A cells. MDA-MB231, however, recover better after 4-days of toxic insult. In addition, nutritional manipulation of media between the cell lines, or pretreatment with penicillamine, did not alter the hormesis effect displayed by MCF-12A. Growth of these cells however was not maintained in the alternative medium. The study demonstrates that a hormesis effect from trace metals is detectable in cultured mammary cells; fluorescent indicators, however, are not as sensitive as cell proliferation or MTT in recognizing the subtle responses. Also, sensitivity of mammary cells to lower concentrations of Cu, a biologically important trace metal, may play an important role in controlling cellular processes and proliferation. The ability to detect this in vitro phenomenon implies that similar processes, occurring in vivo, may be responsible for the development, induction, or enhancement of human cancers.
18 GHz electromagnetic field induces permeability of Gram-positive cocci
Nguyen, The Hong Phong; Shamis, Yury; Croft, Rodney J.; Wood, Andrew; McIntosh, Robert L.; Crawford, Russell J.; Ivanova, Elena P.
2015-01-01
The effect of electromagnetic field (EMF) exposures at the microwave (MW) frequency of 18 GHz, on four cocci, Planococcus maritimus KMM 3738, Staphylococcus aureus CIP 65.8T, S. aureus ATCC 25923 and S. epidermidis ATCC 14990T, was investigated. We demonstrate that exposing the bacteria to an EMF induced permeability in the bacterial membranes of all strains studied, as confirmed directly by transmission electron microscopy (TEM), and indirectly via the propidium iodide assay and the uptake of silica nanospheres. The cells remained permeable for at least nine minutes after EMF exposure. It was shown that all strains internalized 23.5 nm nanospheres, whereas the internalization of the 46.3 nm nanospheres differed amongst the bacterial strains (S. epidermidis ATCC 14990T~ 0%; Staphylococcus aureus CIP 65.8T S. aureus ATCC 25923, ~40%; Planococcus maritimus KMM 3738, ~80%). Cell viability experiments indicated that up to 84% of the cells exposed to the EMF remained viable. The morphology of the bacterial cells was not altered, as inferred from the scanning electron micrographs, however traces of leaked cytosolic fluids from the EMF exposed cells could be detected. EMF-induced permeabilization may represent an innovative, alternative cell permeability technique for applications in biomedical engineering, cell drug delivery and gene therapy. PMID:26077933
A novel optical assay system for the quantitative measurement of chemotaxis.
Kanegasaki, Shiro; Nomura, Yuka; Nitta, Nao; Akiyama, Shuichi; Tamatani, Takuya; Goshoh, Yasuhiro; Yoshida, Takashi; Sato, Tsuyoshi; Kikuchi, Yuji
2003-11-01
We have developed an optically accessible, horizontal chemotaxis apparatus consisting of an etched silicon substrate and a flat glass plate, both of which form two compartments with a 5-microm-deep microchannel in between. The device is held together with a stainless steel holder with holes for injecting cells and a chemoattractant to the different compartments. Migration of cells in the channel is traced with time-lapse intervals using a CCD camera. By developing a method for aligning cells at the edge of the channel, we could successfully reduce the number of cells required for a chemotactic assay, depending on the experiment, to 100 or less. To prevent ceaseless flow of contents between the adjacent compartments via the communicating microchannel, a space at the top end of the holder was filled with medium after aligning the cells. By using a fluorescent probe, we demonstrated experimentally that a stable concentration gradient could be maintained. Furthermore, we determined theoretical details of the gradient established using a model chemokine and a computational fluid dynamics code. Reproducible kinetic results of cell migration were obtained using human neutrophils and IL-8 as a model. Migration of other cells such as eosinophils, basophils and Jurkat lymphocytes toward the appropriate chemokines were also demonstrated.
18 GHz electromagnetic field induces permeability of Gram-positive cocci.
Nguyen, The Hong Phong; Shamis, Yury; Croft, Rodney J; Wood, Andrew; McIntosh, Robert L; Crawford, Russell J; Ivanova, Elena P
2015-06-16
The effect of electromagnetic field (EMF) exposures at the microwave (MW) frequency of 18 GHz, on four cocci, Planococcus maritimus KMM 3738, Staphylococcus aureus CIP 65.8(T), S. aureus ATCC 25923 and S. epidermidis ATCC 14990(T), was investigated. We demonstrate that exposing the bacteria to an EMF induced permeability in the bacterial membranes of all strains studied, as confirmed directly by transmission electron microscopy (TEM), and indirectly via the propidium iodide assay and the uptake of silica nanospheres. The cells remained permeable for at least nine minutes after EMF exposure. It was shown that all strains internalized 23.5 nm nanospheres, whereas the internalization of the 46.3 nm nanospheres differed amongst the bacterial strains (S. epidermidis ATCC 14990(T) ~ 0%; Staphylococcus aureus CIP 65.8(T) S. aureus ATCC 25923, ~40%; Planococcus maritimus KMM 3738, ~ 80%). Cell viability experiments indicated that up to 84% of the cells exposed to the EMF remained viable. The morphology of the bacterial cells was not altered, as inferred from the scanning electron micrographs, however traces of leaked cytosolic fluids from the EMF exposed cells could be detected. EMF-induced permeabilization may represent an innovative, alternative cell permeability technique for applications in biomedical engineering, cell drug delivery and gene therapy.
Harris, Stephen E; Rediske, Michael; Neitzke, Rebecca; Rakian, Audrey
2017-01-01
The periodontium is a complex tissue with epithelial components and a complex set of mesodermal derived alveolar bone, cellular and a cellular cementum, and tendon like ligaments (PDL). The current evidence demonstrates that the major pool of periodontal stem cells is derived from a population of micro vascular associated aSMA-positive stem/progenitor (PSC) cells that by lineage tracing form all three major mesodermal derived components of the periodontium. With in vitro aSMA+ stem cells, transcriptome and chip- seq experiments, the gene network and enhancer maps were determined at several differentiation states of the PSC. Current work on the role of the Bmp2 gene in the periodontal stem cell differentiation demonstrated that this Wnt regulated gene, Bmp2, is necessary for differentiation to all three major mesodermal derived component of the periodontium. The mechanism and use of Sclerostin antibody as an activator of Wnt signaling and Bmp2 gene as a potential route to treat craniofacial bone loss is discussed. As well, the mechanism and use of Pth in the treatment of periodontal bone loss or other craniofacial bone loss is presented in this review. PMID:29457146
Al+Si Interface Optical Properties Obtained in the Si Solar Cell Configuration
Subedi, Indra; Silverman, Timothy J.; Deceglie, Michael G.; ...
2017-10-18
Al is a commonly used material for rear side metallization in commercial silicon (Si) wafer solar cells. In this study, through-the-silicon spectroscopic ellipsometry is used in a test sample to measure Al+Si interface optical properties like those in Si wafer solar cells. Two different spectroscopic ellipsometers are used for measurement of Al+Si interface optical properties over the 1128-2500 nm wavelength range. For validation, the measured interface optical properties are used in a ray tracing simulation over the 300-2500 nm wavelength range for an encapsulated Si solar cell having random pyramidal texture. The ray tracing model matches well with the measuredmore » total reflectance at normal incidence of a commercially available Si module. The Al+Si optical properties presented here enable quantitative assessment of major irradiance/current flux losses arising from reflection and parasitic absorption in encapsulated Si solar cells.« less
Imaging trace element distributions in single organelles and subcellular features
NASA Astrophysics Data System (ADS)
Kashiv, Yoav; Austin, Jotham R.; Lai, Barry; Rose, Volker; Vogt, Stefan; El-Muayed, Malek
2016-02-01
The distributions of chemical elements within cells are of prime importance in a wide range of basic and applied biochemical research. An example is the role of the subcellular Zn distribution in Zn homeostasis in insulin producing pancreatic beta cells and the development of type 2 diabetes mellitus. We combined transmission electron microscopy with micro- and nano-synchrotron X-ray fluorescence to image unequivocally for the first time, to the best of our knowledge, the natural elemental distributions, including those of trace elements, in single organelles and other subcellular features. Detected elements include Cl, K, Ca, Co, Ni, Cu, Zn and Cd (which some cells were supplemented with). Cell samples were prepared by a technique that minimally affects the natural elemental concentrations and distributions, and without using fluorescent indicators. It could likely be applied to all cell types and provide new biochemical insights at the single organelle level not available from organelle population level studies.
Numerical prediction of algae cell mixing feature in raceway ponds using particle tracing methods.
Ali, Haider; Cheema, Taqi A; Yoon, Ho-Sung; Do, Younghae; Park, Cheol W
2015-02-01
In the present study, a novel technique, which involves numerical computation of the mixing length of algae particles in raceway ponds, was used to evaluate the mixing process. A value of mixing length that is higher than the maximum streamwise distance (MSD) of algae cells indicates that the cells experienced an adequate turbulent mixing in the pond. A coupling methodology was adapted to map the pulsating effects of a 2D paddle wheel on a 3D raceway pond in this study. The turbulent mixing was examined based on the computations of mixing length, residence time, and algae cell distribution in the pond. The results revealed that the use of particle tracing methodology is an improved approach to define the mixing phenomenon more effectively. Moreover, the algae cell distribution aided in identifying the degree of mixing in terms of mixing length and residence time. © 2014 Wiley Periodicals, Inc.
Al+Si Interface Optical Properties Obtained in the Si Solar Cell Configuration
DOE Office of Scientific and Technical Information (OSTI.GOV)
Subedi, Indra; Silverman, Timothy J.; Deceglie, Michael G.
Al is a commonly used material for rear side metallization in commercial silicon (Si) wafer solar cells. In this study, through-the-silicon spectroscopic ellipsometry is used in a test sample to measure Al+Si interface optical properties like those in Si wafer solar cells. Two different spectroscopic ellipsometers are used for measurement of Al+Si interface optical properties over the 1128-2500 nm wavelength range. For validation, the measured interface optical properties are used in a ray tracing simulation over the 300-2500 nm wavelength range for an encapsulated Si solar cell having random pyramidal texture. The ray tracing model matches well with the measuredmore » total reflectance at normal incidence of a commercially available Si module. The Al+Si optical properties presented here enable quantitative assessment of major irradiance/current flux losses arising from reflection and parasitic absorption in encapsulated Si solar cells.« less
Dresp-Langley, Birgitta
2011-01-01
Scientific studies have shown that non-conscious stimuli and representations influence information processing during conscious experience. In the light of such evidence, questions about potential functional links between non-conscious brain representations and conscious experience arise. This article discusses neural model capable of explaining how statistical learning mechanisms in dedicated resonant circuits could generate specific temporal activity traces of non-conscious representations in the brain. How reentrant signaling, top-down matching, and statistical coincidence of such activity traces may lead to the progressive consolidation of temporal patterns that constitute the neural signatures of conscious experience in networks extending across large distances beyond functionally specialized brain regions is then explained. PMID:24962683
Song, Chenghui; Ehlers, Vanessa L; Moyer, James R
2015-09-30
Neuronal activity in medial prefrontal cortex (mPFC) is critical for the formation of trace fear memory, yet the cellular mechanisms underlying these memories remain unclear. One possibility involves the modulation of intrinsic excitability within mPFC neurons that project to the basolateral complex of amygdala (BLA). The current study used a combination of retrograde labeling and in vitro whole-cell patch-clamp recordings to examine the effect of trace fear conditioning on the intrinsic excitability of layer 5 mPFC-BLA projection neurons in adult rats. Trace fear conditioning significantly enhanced the intrinsic excitability of regular spiking infralimbic (IL) projection neurons, as evidenced by an increase in the number of action potentials after current injection. These changes were also associated with a reduction in spike threshold and an increase in h current. In contrast, trace fear conditioning reduced the excitability of regular spiking prelimbic (PL) projection neurons, through a learning-related decrease of input resistance. Interestingly, the amount of conditioned freezing was (1) positively correlated with excitability of IL-BLA projection neurons after conditioning and (2) negatively correlated with excitability of PL-BLA projection neurons after extinction. Trace fear conditioning also significantly enhanced the excitability of burst spiking PL-BLA projection neurons. In both regions, conditioning-induced plasticity was learning specific (observed in conditioned but not in pseudoconditioned rats), flexible (reversed by extinction), and transient (lasted <10 d). Together, these data suggest that intrinsic plasticity within mPFC-BLA projection neurons occurs in a subregion- and cell-type-specific manner during acquisition, consolidation, and extinction of trace fear conditioning. Significance statement: Frontal lobe-related function is vital for a variety of important behaviors, some of which decline during aging. This study involves a novel combination of electrophysiological recordings from fluorescently labeled mPFC-to-amygdala projection neurons in rats with acquisition and extinction of trace fear conditioning to determine how specific neurons change during behavior. This is the first study to demonstrate that trace fear conditioning significantly alters the intrinsic excitability of mPFC-to-amygdala projection neurons in a subregion- and cell-type-specific manner, which is also transient and reversed by extinction. These data are of broad interest to the neuroscientific community, and the results will inspire additional studies investigating the cellular mechanisms underlying circuit-specific changes within the brain as a result of associative learning and memory. Copyright © 2015 the authors 0270-6474/15/3513511-14$15.00/0.
NASA Technical Reports Server (NTRS)
Sutton, S. R.
1989-01-01
The Synchrotron X ray Fluorescence (SXRF) microprobe at the National Synchrotron Light Source (NSLS), Brookhaven National Laboratory, will be an excellent instrument for non-destructive trace element analyses of cometary nucleus samples. Trace element analyses of as-received cometary nucleus material will also be possible with this technique. Bulk analysis of relatively volatile elements will be important in establishing comet formation conditions. However, as demonstrated for meteorites, microanalyses of individual phases in their petrographic context are crucial in defining the histories of particular components in unequilibrated specimens. Perhaps most informative in comparing cometary material with meteorites will be the halogens and trace metals. In-situ, high spatial resolution microanalyses will be essential in establishing host phases for these elements and identifying terrestrial (collection/processing) overprints. The present SXRF microprobe is a simple, yet powerful, instrument in which specimens are excited with filtered, continuum synchrotron radiation from a bending magnet on a 2.5 GeV electron storage ring. A refrigerated cell will be constructed to permit analyses at low temperatures. The cell will consist essentially of an air tight housing with a cold stage. Kapton windows will be used to allow the incident synchrotron beam to enter the cell and fluorescent x rays to exit it. The cell will be either under vacuum or continuous purge by ultrapure helium during analyses. Several other improvements of the NSLS microprobe will be made prior to the cometary nucleus sample return mission that will greatly enhance the sensitivity of the technique.
de Gruijter, Madeleine; de Poot, Christianne J; Elffers, Henk
2016-01-01
Currently, a series of promising new tools are under development that will enable crime scene investigators (CSIs) to analyze traces in situ during the crime scene investigation or enable them to detect blood and provide information on the age of blood. An experiment is conducted with thirty CSIs investigating a violent robbery at a mock crime scene to study the influence of such technologies on the perception and interpretation of traces during the first phase of the investigation. Results show that in their search for traces, CSIs are not directed by the availability of technologies, which is a reassuring finding. Qualitative findings suggest that CSIs are generally more focused on analyzing perpetrator traces than on reconstructing the event. A focus on perpetrator traces might become a risk when other crime-related traces are overlooked, and when analyzed traces are in fact not crime-related and in consequence lead to the identification of innocent suspects. © 2015 American Academy of Forensic Sciences.
Both Trace and Delay Conditioning of Evaluative Responses Depend on Contingency Awareness
ERIC Educational Resources Information Center
Kattner, Florian; Ellermeier, Wolfgang; Tavakoli, Paniz
2012-01-01
Whereas previous evaluative conditioning (EC) studies produced inconsistent results concerning the role of contingency knowledge, there are classical eye-blink conditioning studies suggesting that declarative processes are involved in trace conditioning but not in delay conditioning. In two EC experiments pairing neutral sounds (conditioned…
Differential-optoacoustic absorption detector
NASA Technical Reports Server (NTRS)
Shumate, M. S.
1977-01-01
Two-cell spectrophone detects trace amounts of atmospheric pollutants by measuring absorption coefficients of gases with various laser sources. Device measures pressure difference between two tapered cells with differential manometer. Background signal is reduced by balanced window heating and balanced carrier gas absorption in two cells.
NASA Astrophysics Data System (ADS)
Chabot, N. L.
2017-12-01
As planetesimals were heated up in the early Solar System, the formation of Fe-Ni metallic melts was a common occurrence. During planetesimal differentiation, the denser Fe-Ni metallic melts separated from the less dense silicate components, though some meteorites suggest that their parent bodies only experienced partial differentiation. If the Fe-Ni metallic melts did form a central metallic core, the core eventually crystallized to a solid, some of which we sample as iron meteorites. In all of these planetesimal evolution processes, the composition of the Fe-Ni metallic melt influenced the process and the resulting trace element chemical signatures. In particular, the metallic melt's "light element" composition, those elements present in the metallic melt in a significant concentration but with lower atomic masses than Fe, can strongly affect trace element partitioning. Experimental studies have provided critical data to determine the effects of light elements in Fe-Ni metallic melts on trace element partitioning behavior. Here I focus on combining numerous experimental results to identify trace elements that provide unique insight into constraining the light element composition of early Solar System Fe-Ni metallic melts. Experimental studies have been conducted at 1 atm in a variety of Fe-Ni systems to investigate the effects of light elements on trace element partitioning behavior. A frequent experimental examination of the effects of light elements in metallic systems involves producing run products with coexisting solid metal and liquid metal phases. Such solid-metal-liquid-metal experiments have been conducted in the Fe-Ni binary system as well as Fe-Ni systems with S, P, and C. Experiments with O-bearing or Si-bearing Fe-Ni metallic melts do not lend themselves to experiments with coexisting solid metal and liquid metal phases, due to the phase diagrams of these elements, but experiments with two immiscible Fe-Ni metallic melts have provided insight into the qualitative effects of O and Si relative to the well-determined effects of S. Together, these experimental studies provide a robust dataset to identify key elements that are predicted to produce distinct chemical signatures as a function of different Fe-Ni metallic melt compositions during planetesimal evolution processes.
Facilitation of trace metal uptake in cells by inulin coating of metallic nanoparticles
NASA Astrophysics Data System (ADS)
Santillán-Urquiza, Esmeralda; Arteaga-Cardona, Fernando; Torres-Duarte, Cristina; Cole, Bryan; Wu, Bing; Méndez-Rojas, Miguel A.; Cherr, Gary N.
2017-09-01
Trace elements such as zinc and iron are essential for the proper function of biochemical processes, and their uptake and bioavailability are dependent on their chemical form. Supplementation of trace metals through nanostructured materials is a new field, but its application raises concerns regarding their toxicity. Here, we compared the intracellular zinc uptake of different sources of zinc: zinc sulfate, and ZnO and core-shell α-Fe2O3@ZnO nanoparticles, coated or uncoated with inulin, an edible and biocompatible polysaccharide. Using mussel haemocytes, a well-known model system to assess nanomaterial toxicity, we simultaneously assessed zinc accumulation and multiple cellular response endpoints. We found that intracellular zinc uptake was strongly enhanced by inulin coating, in comparison to the uncoated nanoparticles, while no significant effects on cell death, cell viability, mitochondrial membrane integrity, production of reactive oxygen species or lysosome abundance were observed at concentrations up to 20 ppm. Since no significant increments in toxicity were observed, the coated nanomaterials may be useful to increase in vivo zinc uptake for nutritional applications.
The Spindle Cell Neoplasms of the Oral Cavity.
Shamim, Thorakkal
2015-01-01
Spindle cell neoplasms are defined as neoplasms that consist of spindle-shaped cells in the histopathology. Spindle cell neoplasms can affect the oral cavity. In the oral cavity, the origin of the spindle cell neoplasms may be traced to epithelial, mesenchymal and odontogenic components. This article aims to review the spindle cell neoplasms of the oral cavity with emphasis on histopathology.
The Spindle Cell Neoplasms of the Oral Cavity
Shamim, Thorakkal
2015-01-01
Spindle cell neoplasms are defined as neoplasms that consist of spindle-shaped cells in the histopathology. Spindle cell neoplasms can affect the oral cavity. In the oral cavity, the origin of the spindle cell neoplasms may be traced to epithelial, mesenchymal and odontogenic components. This article aims to review the spindle cell neoplasms of the oral cavity with emphasis on histopathology. PMID:26351482
Schausberger, P; Hoffmann, D
2008-11-01
1. Sibling cannibalism is a common phenomenon in the animal kingdom but entails a high risk of direct and inclusive fitness loss for the mother and her offspring. Therefore, mechanisms limiting sibling cannibalism are expected to be selected for. One way of maternal manipulation of sibling cannibalism is to influence hatching asynchrony between nearby laid eggs. This has rarely been tested experimentally. 2. We examined the ability of ovipositing females of the cannibalistic predatory mite Phytoseiulus persimilis to influence the occurrence of sibling cannibalism among offspring by manipulating hatching asynchrony of nearby laid eggs. 3. In the first experiment, we assessed the occurrence of sibling cannibalism in relation to the hatching interval (24 h and 48 h) between nearby laid eggs. In the second experiment, we tested whether ovipositing females discriminate sites containing young (24-h old) and old (48-h old) eggs, fresh and old traces (metabolic waste products and possibly pheromones) left by the same female (24 h and 48 h ago), or young eggs plus fresh female traces and old eggs plus old female traces. Both experiments were conducted with and without prey. 4. Without prey, siblings were more likely to cannibalize each other if the hatching interval between nearby laid eggs was short (24 h). Cannibalism occurred less often when senior siblings (protonymphs) experienced a delay in the opportunity to cannibalize junior siblings (larvae). 5. Independent of prey availability, females preferentially added new eggs to sites containing old eggs plus old female traces but did neither distinguish between young and old eggs presented without own traces nor between fresh and old traces presented without eggs. 6. We discuss cue perception and use by P. persimilis females and contrast the outcome of our experiments and theoretical predictions of sibling cannibalism. We conclude that P. persimilis mothers increase hatching asynchrony of nearby laid eggs to prevent sibling cannibalism on the last produced offspring. Such a behaviour may be considered a simple form of maternal care increasing the survival prospects of offspring.
Mialdun, A; Ryzhkov, I I; Melnikov, D E; Shevtsova, V
2008-08-22
We report experimental evidence of convection caused by translational vibration of nonuniformly heated fluid in low gravity. The theory of vibrational convection in weightlessness is well developed but direct experimental proof has been missing. An innovative point of the experiment is the observation of a temperature field in the front and side views of the cubic cell. In addition, particle tracing is employed. The evolution of this field is studied systematically in a wide range of frequencies and amplitudes. The flow structures reported in previous numerical studies are confirmed. The transition from four-vortex flow to the pattern with three vortices is observed in the transient state.
Staffend, Nancy A; Meisel, Robert L
2011-01-01
Fine neuronal morphology, such as dendritic spines, classically has been studied using the Golgi technique; however, Golgi staining is difficult to combine with other histological techniques. With the increasing popularity of fluorescent imaging, a number of fluorescent dyes have been developed that enable the coupling of multiple fluorescent labels in a single preparation. These fluorescent dyes include the lipophilic dialkylcarbocyanine, DiI; traditionally used for anterograde and retrograde neuronal tracing. More recently, DiI labeling has been used in combination with the Gene Gun for "DiOlistic" labeling of neurons in slice preparations. DiI sequesters itself within and diffuses laterally along the neuronal membrane, however once the cell is permeabilized, the DiI begins to leak from the cell membrane. A DiI derivative, Cell Tracker™ CM-DiI, increases dye stability and labeling half-life in permeabilized tissue, however at much greater expense. Here, the DiI and CM-DiI DiOlistic labeling techniques were tested in side-by-side experiments evaluating dye stability within dendritic architecture in medium spiny neurons of the dorsal stratum in both non-permeabilized and permeabilized tissue sections. In tissue sections that were not permeabilized, spine density in DiI labeled sections was higher than in CM-DiI labeling. In contrast, tissue sections that were permeabilized had higher spine densities in CM-DiI labeled neurons. These results suggest that for experiments involving non-permeabilized tissue, traditional DiI will suffice, however for experiments involving permeabilized tissue CM-DiI provides more consistent data. These experiments provide the first quantitative analyses of the impact of methodological permutations on neuronal labeling with DiI.
Retracing in correlative light electron microscopy: where is my object of interest?
Hodgson, Lorna; Nam, David; Mantell, Judith; Achim, Alin; Verkade, Paul
2014-01-01
Correlative light electron microscopy (CLEM) combines the strengths of light and electron microscopy in a single experiment. There are many ways to perform a CLEM experiment and a variety of microscopy modalities can be combined either on separate instruments or as completely integrated solutions. In general, however, a CLEM experiment can be divided into three parts: probes, processing, and analysis. Most of the existing technologies are focussed around the development and use of probes or describe processing methodologies that explain or circumvent some of the compromises that need to be made when performing both light and electron microscopy on the same sample. So far, relatively little attention has been paid to the analysis part of CLEM experiments. Although it is an essential part of each CLEM experiment, it is usually a cumbersome manual process. Here, we briefly discuss each of the three above-mentioned steps, with a focus on the analysis part. We will also introduce an automated registration algorithm that can be applied to the analysis stage to enable the accurate registration of LM and EM images. This facilitates tracing back the right cell/object seen in the light microscope in the EM. © 2014 Elsevier Inc. All rights reserved.
Objects of attention, objects of perception.
Avrahami, J
1999-11-01
Four experiments were conducted, to explore the notion of objects in perception. Taking as a starting point the effects of display content on rapid attention transfer and manipulating curvature, closure, and processing time, a link between objects of attention and objects of perception is proposed. In Experiment 1, a number of parallel, equally spaced, straight lines facilitated attention transfer along the lines, relative to transfer across the lines. In Experiment 2, with curved, closed-contour shapes, no "same-object" facilitation was observed. However, when a longer time interval was provided, in Experiment 3, a same-object advantage started to emerge. In Experiment 4, using the same curved shapes but in a non-speeded distance estimation task, a strong effect of objects was observed. It is argued that attention transfer is facilitated by line tracing but that line tracing is encouraged by objects.
Fast first arrival picking algorithm for noisy microseismic data
NASA Astrophysics Data System (ADS)
Kim, Dowan; Byun, Joongmoo; Lee, Minho; Choi, Jihoon; Kim, Myungsun
2017-01-01
Most microseismic events occur during hydraulic fracturing. Thus microseismic monitoring, by recording seismic waves from microseismic events, is one of the best methods for locating the positions of hydraulic fractures. However, since microseismic events have very low energy, the data often have a low signal-to-noise ratio (S/N ratio) and it is not easy to pick the first arrival time. In this study, we suggest a new fast picking method optimised for noisy data using cross-correlation and stacking. In this method, a reference trace is selected and the time differences between the first arrivals of the reference trace and those of the other traces are computed by cross-correlation. Then, all traces are aligned with the reference trace by time shifting, and the aligned traces are summed together to produce a stacked reference trace that has a considerably improved S/N ratio. After the first arrival time of the stacked reference trace is picked, the first arrival time of each trace is calculated automatically using the time differences obtained in the cross-correlation process. In experiments with noisy synthetic data and field data, this method produces more reliable results than the traditional method, which picks the first arrival time of each noisy trace separately. In addition, the computation time is dramatically reduced.
Assessing Students' Ability to Trace Matter in Dynamic Systems in Cell Biology
ERIC Educational Resources Information Center
Wilson, Christopher D.; Anderson, Charles W.; Heidemann, Merle; Merrill, John E.; Merritt, Brett W.; Richmond, Gail; Sibley, Duncan F.; Parker, Joyce M.
2006-01-01
College-level biology courses contain many complex processes that are often taught and learned as detailed narratives. These processes can be better understood by perceiving them as dynamic systems that are governed by common fundamental principles. Conservation of matter is such a principle, and thus tracing matter is an essential step in…
NASA Astrophysics Data System (ADS)
Abu-Taha, M. I.; Abu-Teir, M. M.; Al-Jamal, A. J.; Eideh, H.
The aim of this work was to establish the feasibility of the combined photoacoustic (PA) and photopyroelectric (PPE) detection of the vapours emitted from essential oils and their corresponding uncrushed leaves or flowers. Gas traces of jasmine (Jessamine (Jasminum)), mint (Mentha arvensis L.) and Damask rose (Rosa damascena Miller) and their essential oils were tested using a combined cell fitted with both a photopyroelectric film (PVDF) and a microphone in conjunction with a pulsed wideband infrared source (PWBS) source. Infrared PA and PPE absorbances were obtained simultaneously at room temperatures with excellent reproducibility and high signal-to-noise ratios. Significant similarities found between the PA and PPE spectra of the trace gas emissions of plant parts, i.e., flowers or leaves and their related essential oils show the good correlation of their emissions and that both effects are initiated by the same absorbing molecules.
Quantification of Crypt and Stem Cell Evolution in the Normal and Neoplastic Human Colon
Baker, Ann-Marie; Cereser, Biancastella; Melton, Samuel; Fletcher, Alexander G.; Rodriguez-Justo, Manuel; Tadrous, Paul J.; Humphries, Adam; Elia, George; McDonald, Stuart A.C.; Wright, Nicholas A.; Simons, Benjamin D.; Jansen, Marnix; Graham, Trevor A.
2014-01-01
Summary Human intestinal stem cell and crypt dynamics remain poorly characterized because transgenic lineage-tracing methods are impractical in humans. Here, we have circumvented this problem by quantitatively using somatic mtDNA mutations to trace clonal lineages. By analyzing clonal imprints on the walls of colonic crypts, we show that human intestinal stem cells conform to one-dimensional neutral drift dynamics with a “functional” stem cell number of five to six in both normal patients and individuals with familial adenomatous polyposis (germline APC−/+). Furthermore, we show that, in adenomatous crypts (APC−/−), there is a proportionate increase in both functional stem cell number and the loss/replacement rate. Finally, by analyzing fields of mtDNA mutant crypts, we show that a normal colon crypt divides around once every 30–40 years, and the division rate is increased in adenomas by at least an order of magnitude. These data provide in vivo quantification of human intestinal stem cell and crypt dynamics. PMID:25127143
Live cell imaging reveals marked variability in myoblast proliferation and fate
2013-01-01
Background During the process of muscle regeneration, activated stem cells termed satellite cells proliferate, and then differentiate to form new myofibers that restore the injured area. Yet not all satellite cells contribute to muscle repair. Some continue to proliferate, others die, and others become quiescent and are available for regeneration following subsequent injury. The mechanisms that regulate the adoption of different cell fates in a muscle cell precursor population remain unclear. Methods We have used live cell imaging and lineage tracing to study cell fate in the C2 myoblast line. Results Analyzing the behavior of individual myoblasts revealed marked variability in both cell cycle duration and viability, but similarities between cells derived from the same parental lineage. As a consequence, lineage sizes and outcomes differed dramatically, and individual lineages made uneven contributions toward the terminally differentiated population. Thus, the cohort of myoblasts undergoing differentiation at the end of an experiment differed dramatically from the lineages present at the beginning. Treatment with IGF-I increased myoblast number by maintaining viability and by stimulating a fraction of cells to complete one additional cell cycle in differentiation medium, and as a consequence reduced the variability of the terminal population compared with controls. Conclusion Our results reveal that heterogeneity of responses to external cues is an intrinsic property of cultured myoblasts that may be explained in part by parental lineage, and demonstrate the power of live cell imaging for understanding how muscle differentiation is regulated. PMID:23638706
Human Muse Cells Reconstruct Neuronal Circuitry in Subacute Lacunar Stroke Model.
Uchida, Hiroki; Niizuma, Kuniyasu; Kushida, Yoshihiro; Wakao, Shohei; Tominaga, Teiji; Borlongan, Cesario V; Dezawa, Mari
2017-02-01
Multilineage-differentiating stress-enduring (muse) cells are endogenous nontumorigenic stem cells with pluripotency harvestable as pluripotent marker SSEA-3 + cells from the bone marrow from cultured bone marrow-mesenchymal stem cells. After transplantation into neurological disease models, muse cells exert repair effects, but the exact mechanism remains inconclusive. We conducted mechanism-based experiments by transplanting serum/xeno-free cultured-human bone marrow-muse cells into the perilesion brain at 2 weeks after lacunar infarction in immunodeficient mice. Approximately 28% of initially transplanted muse cells remained in the host brain at 8 weeks, spontaneously differentiated into cells expressing NeuN (≈62%), MAP2 (≈30%), and GST-pi (≈12%). Dextran tracing revealed connections between host neurons and muse cells at the lesioned motor cortex and the anterior horn. Muse cells extended neurites through the ipsilateral pyramidal tract, crossed to contralateral side, and reached to the pyramidal tract in the dorsal funiculus of spinal cord. Muse-transplanted stroke mice displayed significant recovery in cylinder tests, which was reverted by the human-selective diphtheria toxin. At 10 months post-transplantation, human-specific Alu sequence was detected only in the brain but not in other organs, with no evidence of tumor formation. Transplantation at the delayed subacute phase showed muse cells differentiated into neural cells, facilitated neural reconstruction, improved functions, and displayed solid safety outcomes over prolonged graft maturation period, indicating their therapeutic potential for lacunar stroke. © 2016 The Authors.
Qamar, Wajhul; Al-Ghadeer, Abdul Rahman; Ali, Raisuddin; Abuelizz, Hatem A
2017-08-01
The main objective was to determine the elemental profile of the lung lining fluid of rats which are used as model animals in various experiments. Lung lining fluid elemental constitution obtained after bronchoalveolar lavage fluid (BALF) was analyzed by inductively coupled plasma mass spectrometry (ICP-MS) to determine the biological trace elements along with calcium and magnesium. BALF was collected from healthy rats using a tracheal cannula. However, cells in BALF were counted to monitor any underlying inflammatory lung condition. Cell free BALF samples were processed and analyzed for the elements including magnesium (Mg), calcium (Ca), chromium (Cr), manganese (Mn), iron (Fe), nickel (Ni), copper (Cu), zinc (Zn), selenium (Se), bromine (Br), and iodine (I). In view of this, calcium concentration was the highest (6318.08 ± 3094.3 μg/L) and copper concentration was the lowest (0.89 ± 0.21 μg/L). The detected elements, from high to low concentration, include Ca > Mg > Fe > Br > I > Cr > Ni > Zn > Mn > Se > Cu. Pearson's correlation analysis revealed no significant correlation between cell count and concentration of any of the element detected in BALF. Correlation analysis also revealed significant positive correlation among Fe, I, Cr, Ni, and Mn. Ca was found to be correlated negatively with Cu and positively with Se. Br and Mg found to be positively correlated with each other. Zn remained the only element that was not found to be correlated with any of the elements in the rat BALF.
From non-random molecular structure to life and mind
NASA Technical Reports Server (NTRS)
Fox, S. W.
1989-01-01
The evolutionary hierarchy molecular structure-->macromolecular structure-->protobiological structure-->biological structure-->biological functions has been traced by experiments. The sequence always moves through protein. Extension of the experiments traces the formation of nucleic acids instructed by proteins. The proteins themselves were, in this picture, instructed by the self-sequencing of precursor amino acids. While the sequence indicated explains the thread of the emergence of life, protein in cellular membrane also provides the only known material basis for the emergence of mind in the context of emergence of life.
KLF4-dependent perivascular cell plasticity mediates pre-metastatic niche formation and metastasis
Murgai, Meera; Ju, Wei; Eason, Matthew; Kline, Jessica; Beury, Daniel; Kaczanowska, Sabina; Miettinen, Markku M; Kruhlak, Michael; Lei, Haiyan; Shern, Jack F; Cherepanova, Olga A.; Owens, Gary K; Kaplan, Rosandra N
2017-01-01
A deeper understanding of the metastatic process is required for the development of new therapies that improve patient survival. Metastatic tumor cell growth and survival in distant organs is facilitated by the formation of a pre-metastatic niche composed of hematopoietic cells, stromal cells, and extracellular matrix (ECM). Perivascular cells, including vascular smooth muscle cells (vSMCs) and pericytes, are involved in new vessel formation and in promoting stem cell maintenance and proliferation. Given the well-described plasticity of perivascular cells, we hypothesize that perivascular cells similarly regulate tumor cell fate at metastatic sites. Using perivascular cell-specific and pericyte-specific lineage-tracing models, we trace the fate of perivascular cells in the pre-metastatic and metastatic microenvironments. We show that perivascular cells lose the expression of traditional vSMC/pericyte markers in response to tumor-secreted factors and exhibit increased proliferation, migration, and ECM synthesis. Increased expression of the pluripotency gene Klf4 in these phenotypically-switched perivascular cells promotes a less differentiated state characterized by enhanced ECM production that establishes a pro-metastatic fibronectin-rich environment. Genetic inactivation of Klf4 in perivascular cells decreases pre-metastatic niche formation and metastasis. Our data reveal a previously unidentified role for perivascular cells in pre-metastatic niche formation and uncover novel strategies for limiting metastasis. PMID:28920957
The effects of handwriting experience on functional brain development in pre-literate children
James, Karin H.; Engelhardt, Laura
2014-01-01
In an age of increasing technology, the possibility that typing on a keyboard will replace handwriting raises questions about the future usefulness of handwriting skills. Here we present evidence that brain activation during letter perception is influenced in different, important ways by previous handwriting of letters versus previous typing or tracing of those same letters. Preliterate, five-year old children printed, typed, or traced letters and shapes, then were shown images of these stimuli while undergoing functional MRI scanning. A previously documented “reading circuit” was recruited during letter perception only after handwriting—not after typing or tracing experience. These findings demonstrate that handwriting is important for the early recruitment in letter processing of brain regions known to underlie successful reading. Handwriting therefore may facilitate reading acquisition in young children. PMID:25541600
EWS/ATF1 expression induces sarcomas from neural crest–derived cells in mice
Yamada, Kazunari; Ohno, Takatoshi; Aoki, Hitomi; Semi, Katsunori; Watanabe, Akira; Moritake, Hiroshi; Shiozawa, Shunichi; Kunisada, Takahiro; Kobayashi, Yukiko; Toguchida, Junya; Shimizu, Katsuji; Hara, Akira; Yamada, Yasuhiro
2013-01-01
Clear cell sarcoma (CCS) is an aggressive soft tissue malignant tumor characterized by a unique t(12;22) translocation that leads to the expression of a chimeric EWS/ATF1 fusion gene. However, little is known about the mechanisms underlying the involvement of EWS/ATF1 in CCS development. In addition, the cellular origins of CCS have not been determined. Here, we generated EWS/ATF1-inducible mice and examined the effects of EWS/ATF1 expression in adult somatic cells. We found that forced expression of EWS/ATF1 resulted in the development of EWS/ATF1-dependent sarcomas in mice. The histology of EWS/ATF1-induced sarcomas resembled that of CCS, and EWS/ATF1-induced tumor cells expressed CCS markers, including S100, SOX10, and MITF. Lineage-tracing experiments indicated that neural crest–derived cells were subject to EWS/ATF1-driven transformation. EWS/ATF1 directly induced Fos in an ERK-independent manner. Treatment of human and EWS/ATF1-induced CCS tumor cells with FOS-targeted siRNA attenuated proliferation. These findings demonstrated that FOS mediates the growth of EWS/ATF1-associated sarcomas and suggest that FOS is a potential therapeutic target in human CCS. PMID:23281395
Shashaani, Hani; Faramarzpour, Mahsa; Hassanpour, Morteza; Namdar, Nasser; Alikhani, Alireza; Abdolahad, Mohammad
2016-11-15
Electrochemical approaches have played crucial roles in bio sensing because of their Potential in achieving sensitive, specific and low-cost detection of biomolecules and other bio evidences. Engineering the electrochemical sensing interface with nanomaterials tends to new generations of label-free biosensors with improved performances in terms of sensitive area and response signals. Here we applied Silicon Nanowire (SiNW) array electrodes (in an integrated architecture of working, counter and reference electrodes) grown by low pressure chemical vapor deposition (LPCVD) system with VLS procedure to electrochemically diagnose the presence of breast cancer cells as well as their response to anticancer drugs. Mebendazole (MBZ), has been used as antitubulin drug. It perturbs the anodic/cathodic response of the cell covered biosensor by releasing Cytochrome C in cytoplasm. Reduction of cytochrome C would change the ionic state of the cells monitored by SiNW biosensor. By applying well direct bioelectrical contacts with cancer cells, SiNWs can detect minor signal transduction and bio recognition events, resulting in precise biosensing. Our device detected the trace of MBZ drugs (with the concentration of 2nM) on electrochemical activity MCF-7 cells. Also, experimented biological analysis such as confocal and Flowcytometry assays confirmed the electrochemical results. Copyright © 2016 Elsevier B.V. All rights reserved.
Pan, Wei-Xing; Schmidt, Robert; Wickens, Jeffery R; Hyland, Brian I
2005-06-29
Behavioral conditioning of cue-reward pairing results in a shift of midbrain dopamine (DA) cell activity from responding to the reward to responding to the predictive cue. However, the precise time course and mechanism underlying this shift remain unclear. Here, we report a combined single-unit recording and temporal difference (TD) modeling approach to this question. The data from recordings in conscious rats showed that DA cells retain responses to predicted reward after responses to conditioned cues have developed, at least early in training. This contrasts with previous TD models that predict a gradual stepwise shift in latency with responses to rewards lost before responses develop to the conditioned cue. By exploring the TD parameter space, we demonstrate that the persistent reward responses of DA cells during conditioning are only accurately replicated by a TD model with long-lasting eligibility traces (nonzero values for the parameter lambda) and low learning rate (alpha). These physiological constraints for TD parameters suggest that eligibility traces and low per-trial rates of plastic modification may be essential features of neural circuits for reward learning in the brain. Such properties enable rapid but stable initiation of learning when the number of stimulus-reward pairings is limited, conferring significant adaptive advantages in real-world environments.
Wang, W.-X.; Fisher, N.S.; Luoma, S.N.
1995-01-01
Pulse-chase feeding and multi-labeled radiotracer techniques were employed to measure the assimilation of 6 trace elements (110mAg, 241Am, 109Cd, 57Co, 75Se and 65Zn) from ingested diatoms in the mussel Mytilus edulis feeding at different rates (0.1, 0.49 and 1.5 mg dry wt h-1). Uniformly radiolabeled diatoms Thalassiosira pseudonana were fed to mussels for 0.5 h, and the behavior of the radiotracers in individual mussels was followed for 96 h in a depuration seawater system. Assimilation efficiency (AE) of each element declined with increasing ingestion rate and increased with gut passage time. The importance of extracellular digestion relative to intracellular digestion increased with ingestion activity, which, when coupled with a decline in AE, suggested that extracellular digestion is less efficient in metal absorption. Zn assimilation was most affected by ingestion rate, suggesting that AE may play a role in the physiological regulation of this metal in M. edulis. In an experiment to simulate the effects of an acidic gut, lowered pH (5.5) enhanced the release of elements from intact diatom cells, especially at low particle concentration. These results indicate that both feeding components of the mussel (i.e. gut passage time, digestive partitioning) and metal chemistry (i.e. metal release at lowered pH within the bivalve gut) are responsible for the difference in the assimilation of trace metals at different food quantities observed in mussels.
Jafri, Azliana Jusnida Ahmad; Arfuzir, Natasha Najwa Nor; Lambuk, Lidawani; Iezhitsa, Igor; Agarwal, Renu; Agarwal, Puneet; Razali, Norhafiza; Krasilnikova, Anna; Kharitonova, Maria; Demidov, Vasily; Serebryansky, Evgeny; Skalny, Anatoly; Spasov, Alexander; Yusof, Ahmad Pauzi Md; Ismail, Nafeeza Mohd
2017-01-01
Glutamate-mediated excitotoxicity involving N-methyl-d-aspartate (NMDA) receptors has been recognized as a final common outcome in pathological conditions involving death of retinal ganglion cells (RGCs). Overstimulation of NMDA receptors results in influx of calcium (Ca) and sodium (Na) ions and efflux of potassium (K). NMDA receptors are blocked by magnesium (Mg). Such changes due to NMDA overstimulation are also associated with not only the altered levels of minerals but also that of trace elements and redox status. Both the decreased and elevated levels of trace elements such as iron (Fe), zinc (Zn), copper (Cu) affect NMDA receptor excitability and redox status. Manganese (Mn), and selenium (Se) are also part of antioxidant defense mechanisms in retina. Additionally endogenous substances such as taurine also affect NMDA receptor activity and retinal redox status. Therefore, the aim of this study was to evaluate the effect of Mg acetyltaurate (MgAT) on the retinal mineral and trace element concentration, oxidative stress, retinal morphology and retinal cell apoptosis in rats after-NMDA exposure. One group of Sprague Dawley rats received intravitreal injection of vehicle while 4 other groups similarly received NMDA (160nmolL -1 ). Among the NMDA injected groups, 3 groups also received MgAT (320nmolL -1 ) as pre-treatment, co-treatment or post-treatment. Seven days after intravitreal injection, rats were sacrificed, eyes were enucleated and retinae were isolated for estimation of mineral (Ca, Na, K, Mg) and trace element (Mn, Cu, Fe, Se, Zn) concentration using Inductively Coupled Plasma (DRC ICP-MS) techniques (NexION 300D), retinal oxidative stress using Elisa, retinal morphology using H&E staining and retinal cell apoptosis using terminal deoxynucleotidyl transferase dUTP nick-end labeling (TUNEL). Intravitreal NMDA injection resulted in increased concentration of Ca (4.6 times, p<0.0001), Mg (1.5 times, p<0.01), Na (3 times, p<0.0001) and K (2.3 times, p<0.0001) compared to vehicle injected group. This was accompanied with significant increase of Ca/Mg and Na/K ratios, 3 and 1.27 times respectively, compared to control group. The trace elements such as Cu, Fe and Zn also showed a significant increase amounting to 3.3 (p<0.001), 2.3 (p<0.0001) and 3 (p<0.0001) times respectively compared to control group. Se was increased by 60% (p<0.005). Pre-treatment with MgAT abolished effect of NMDA on minerals and trace elements more effectively than co- and post-treatment. Similar observations were made for retinal oxidative stress, retinal morphology and retinal cell apoptosis. In conclusion, current study demonstrated the protective effect of MgAT against NMDA-induced oxidative stress and retinal cell apoptosis. This effect of MgAT was associated with restoration of retinal concentrations of minerals and trace elements. Further studies are warranted to explore the precise molecular targets of MgAT. Nevertheless, MgAT seems a potential candidate in the management of diseases involving NMDA-induced excitotoxicity. Copyright © 2016 Elsevier GmbH. All rights reserved.
Shape Recognition in Infancy: Visual Integration of Sequential Information.
ERIC Educational Resources Information Center
Rose, Susan A
1988-01-01
Investigated infants' integration of visual information across space and time. In four experiments, infants aged 12 months and 6 months viewed objects after watching light trace similar and dissimilar shapes. Infants looked longer at novel shapes, although six-month-olds did not recognize figures taking more than 10 seconds to trace. One-year-old…
A Forensic Experiment: The Case of the Crime at the Cinema
ERIC Educational Resources Information Center
Valente Nabais, J. M.; Costa, Sara D.
2017-01-01
This paper reports an experimental activity where students have to carefully analyze the evidence collected at the crime scene, namely fibers and lipstick traces. The fibers are analyzed by infrared spectroscopy, solubility tests, and optical microscopy, while in turn the lipstick traces are investigated by thin layer chromatography. Students also…
ERIC Educational Resources Information Center
Snipes, Katherine H.
2009-01-01
A set of computer-based recreation choice experiments were run to examine the effect of expected congestion and social interactions on the decision making process. MouseTrace is a process-tracing program that recorded individual subject's information acquisitions and provided the necessary information to determine if subjects used attribute-based…
ERIC Educational Resources Information Center
Lavilla, Isela; Costas, Marta; Pena-Pereira, Francisco; Gil, Sandra; Bendicho, Carlos
2011-01-01
Ultrasound-assisted extraction (UAE) is introduced to upper-level analytical chemistry students as a simple strategy focused on sample preparation for trace-metal determination in biological tissues. Nickel extraction in seafood samples and quantification by electrothermal atomic absorption spectrometry (ETAAS) are carried out by a team of four…
Decay uncovered in nonverbal short-term memory.
Mercer, Tom; McKeown, Denis
2014-02-01
Decay theory posits that memory traces gradually fade away over the passage of time unless they are actively rehearsed. Much recent work exploring verbal short-term memory has challenged this theory, but there does appear to be evidence for trace decay in nonverbal auditory short-term memory. Numerous discrimination studies have reported a performance decline as the interval separating two tones is increased, consistent with a decay process. However, most of this tone comparison research can be explained in other ways, without reference to decay, and these alternative accounts were tested in the present study. In Experiment 1, signals were employed toward the end of extended retention intervals to ensure that listeners were alert to the presence and frequency content of the memoranda. In Experiment 2, a mask stimulus was employed in an attempt to distinguish between a highly detailed sensory trace and a longer-lasting short-term memory, and the distinctiveness of the stimuli was varied. Despite these precautions, slow-acting trace decay was observed. It therefore appears that the mere passage of time can lead to forgetting in some forms of short-term memory.
Trace Metal-Humic Complexes in Natural Waters: Insights From Speciation Experiments
NASA Astrophysics Data System (ADS)
Stern, J. C.; Salters, V.; Sonke, J.
2006-12-01
The DOM cycle is intimately linked to the cycling and bioavailability of trace metals in aqueous environments. The presence or absence of DOM in the water column can determined whether trace elements will be present in limited quantities as a nutrient, or in surplus quantities as a toxicant. Humic substances (HS), which represent the refractory products of DOM degradation, strongly affect the speciation of trace metals in natural waters. To simulate metal-HS interactions in nature, experiments must be carried out using trace metal concentrations. Sensitive detection systems such as ICP-MS make working with small (nanomolar) concentrations possible. Capillary electrophoresis coupled with ICP-MS (CE-ICP-MS) has recently been identified as a rapid and accurate method to separate metal species and calculate conditional binding constants (log K_c) of metal-humic complexes. CE-ICP-MS was used to measure partitioning of metals between humic substances and a competing ligand (EDTA) and calculate binding constants of rare earth element (REE) and Th, Hf, and Zr-humic complexes at pH 3.5-8 and ionic strength of 0.1. Equilibrium dialysis ligand exchange (EDLE) experiments to validate the CE-ICP-MS method were performed to separate the metal-HS and metal-EDTA species by partitioning due to size exclusion via diffusion through a 1000 Da membrane. CE-ICP-MS experiments were also conducted to compare binding constants of REE with humic substances of various origin, including soil, peat, and aquatic DOM. Results of our experiments show an increase in log K_c with decrease in ionic radius for REE-humic complexes (the lanthanide contraction effect). Conditional binding constants of tetravalent metal-humic complexes were found to be several orders of magnitude higher than REE-humic complexes, indicating that tetravalent metals have a very strong affinity for humic substances. Because thorium is often used as a proxy for the tetravalent actinides, Th-HS binding constants can allow us to assess the importance of tetravalent actinide-humic complexes in groundwater transport from nuclear repositories. Our results suggest that tetravalent actinide-humic complexes couild be more important to account for in predictive speciation models than previously thought.
DOE Office of Scientific and Technical Information (OSTI.GOV)
Masson, H.A.; Denholm, A.M.; Ling, J.R.
1991-06-01
Cells of Bacillus megaterium GW1 and Escherichia coli W7-M5 were specifically radiolabeled with 2,2{prime}-diamino (G-{sup 3}H) pimelic acid (({sup 3}H)DAP) as models of gram-positive and gram-negative bacteria, respectively. Two experiments were conducted to study the in vivo metabolism of 2,2{prime}-diaminopimelic acid (DAP) in sheep. In experiment 1, cells of ({sup 3}H)DAP-labeled B. megaterium GW1 were infused into the rumen of one sheep and the radiolabel was traced within microbial samples, digesta, and the whole animal. Bacterially bound ({sup 3}H)DAP was extensively metabolized, primarily (up to 70% after 8 h) via decarboxylation to ({sup 3}H)lysine by both ruminal protozoa and ruminalmore » bacteria. Recovery of infused radiolabel in urine and feces was low (42% after 96 h) and perhaps indicative of further metabolism by the host animal. In experiment 2, ({sup 3}H)DAP-labeled B. megaterium GW1 was infused into the rumens of three sheep and ({sup 3}H)DAP-labeled E. coli W7-W5 was infused into the rumen of another sheep. The radioactivity contents of these mutant bacteria were insufficient to use as tracers, but the metabolism of DAP was monitored in the total, free, and peptidyl forms. Free DAP, as a proportion of total DPA in duodenal digesta, varied from 0 to 9.5%, whereas peptidyl DAP accounted for 8.3 to 99.2%.« less
RAY-RAMSES: a code for ray tracing on the fly in N-body simulations
DOE Office of Scientific and Technical Information (OSTI.GOV)
Barreira, Alexandre; Llinares, Claudio; Bose, Sownak
2016-05-01
We present a ray tracing code to compute integrated cosmological observables on the fly in AMR N-body simulations. Unlike conventional ray tracing techniques, our code takes full advantage of the time and spatial resolution attained by the N-body simulation by computing the integrals along the line of sight on a cell-by-cell basis through the AMR simulation grid. Moroever, since it runs on the fly in the N-body run, our code can produce maps of the desired observables without storing large (or any) amounts of data for post-processing. We implemented our routines in the RAMSES N-body code and tested the implementationmore » using an example of weak lensing simulation. We analyse basic statistics of lensing convergence maps and find good agreement with semi-analytical methods. The ray tracing methodology presented here can be used in several cosmological analysis such as Sunyaev-Zel'dovich and integrated Sachs-Wolfe effect studies as well as modified gravity. Our code can also be used in cross-checks of the more conventional methods, which can be important in tests of theory systematics in preparation for upcoming large scale structure surveys.« less
Dieltjes, Patrick; Mieremet, René; Zuniga, Sofia; Kraaijenbrink, Thirsa; Pijpe, Jeroen; de Knijff, Peter
2011-07-01
Exploring technological limits is a common practice in forensic DNA research. Reliable genetic profiling based on only a few cells isolated from trace material retrieved from a crime scene is nowadays more and more the rule rather than the exception. On many crime scenes, cartridges, bullets, and casings (jointly abbreviated as CBCs) are regularly found, and even after firing, these potentially carry trace amounts of biological material. Since 2003, the Forensic Laboratory for DNA Research is routinely involved in the forensic investigation of CBCs in the Netherlands. Reliable DNA profiles were frequently obtained from CBCs and used to match suspects, victims, or other crime scene-related DNA traces. In this paper, we describe the sensitive method developed by us to extract DNA from CBCs. Using PCR-based genotyping of autosomal short tandem repeats, we were able to obtain reliable and reproducible DNA profiles in 163 out of 616 criminal cases (26.5%) and in 283 out of 4,085 individual CBC items (6.9%) during the period January 2003-December 2009. We discuss practical aspects of the method and the sometimes unexpected effects of using cell lysis buffer on the subsequent investigation of striation patterns on CBCs.
Lgr5(+ve) stem/progenitor cells contribute to nephron formation during kidney development.
Barker, Nick; Rookmaaker, Maarten B; Kujala, Pekka; Ng, Annie; Leushacke, Marc; Snippert, Hugo; van de Wetering, Marc; Tan, Shawna; Van Es, Johan H; Huch, Meritxell; Poulsom, Richard; Verhaar, Marianne C; Peters, Peter J; Clevers, Hans
2012-09-27
Multipotent stem cells and their lineage-restricted progeny drive nephron formation within the developing kidney. Here, we document expression of the adult stem cell marker Lgr5 in the developing kidney and assess the stem/progenitor identity of Lgr5(+ve) cells via in vivo lineage tracing. The appearance and localization of Lgr5(+ve) cells coincided with that of the S-shaped body around embryonic day 14. Lgr5 expression remained restricted to cell clusters within developing nephrons in the cortex until postnatal day 7, when expression was permanently silenced. In vivo lineage tracing identified Lgr5 as a marker of a stem/progenitor population within nascent nephrons dedicated to generating the thick ascending limb of Henle's loop and distal convoluted tubule. The Lgr5 surface marker and experimental models described here will be invaluable for deciphering the contribution of early nephron stem cells to developmental defects and for isolating human nephron progenitors as a prerequisite to evaluating their therapeutic potential. Copyright © 2012 The Authors. Published by Elsevier Inc. All rights reserved.
Van Keymeulen, Alexandra; Fioramonti, Marco; Centonze, Alessia; Bouvencourt, Gaëlle; Achouri, Younes; Blanpain, Cédric
2017-08-15
The mammary gland (MG) is composed of different cell lineages, including the basal and the luminal cells (LCs) that are maintained by distinct stem cell (SC) populations. LCs can be subdivided into estrogen receptor (ER) + and ER - cells. LCs act as the cancer cell of origin in different types of mammary tumors. It remains unclear whether the heterogeneity found in luminal-derived mammary tumors arises from a pre-existing heterogeneity within LCs. To investigate LC heterogeneity, we used lineage tracing to assess whether the ER + lineage is maintained by multipotent SCs or by lineage-restricted SCs. To this end, we generated doxycycline-inducible ER-rtTA mice that allowed us to perform genetic lineage tracing of ER + LCs and study their fate and long-term maintenance. Our results show that ER + cells are maintained by lineage-restricted SCs that exclusively contribute to the expansion of the ER + lineage during puberty and their maintenance during adult life. Copyright © 2017 The Author(s). Published by Elsevier Inc. All rights reserved.
Stem cell derived endochondral cartilage stimulates bone healing by tissue transformation
Bahney, Chelsea S; Hu, Diane P; Taylor, Aaron J; Ferro, Federico; Britz, Hayley M; Hallgrimsson, Benedikt; Johnstone, Brian; Miclau, Theodore; Marcucio, Ralph S
2016-01-01
Although bone has great capacity for repair, there are a number of clinical situations (fracture non-unions, spinal fusions, revision arthroplasty, segmental defects) in which auto- or allografts augment bone regeneration. Critical failures associated with current grafting treatments include osteonecrosis and limited integration between graft and host tissue. We speculated that the underlying problem with current bone grafting techniques is that they promote bone regeneration through direct osteogenesis. We hypothesized that using cartilage to promote endochondral bone regeneration would leverage normal developmental and repair sequences to produce a well-vascularized regenerate that integrates with the host tissue. In this study we use a translational murine model of a segmental tibia defect to test the clinical utility of bone regeneration from a cartilage graft. We further test the mechanism by which cartilage promotes bone regeneration using in vivo lineage tracing and in vitro culture experiments. Our data show that cartilage grafts support regeneration of a vascularized and integrated bone tissue in vivo, and subsequently propose a translational tissue engineering platform using chondrogenesis of MSCs. Interestingly, lineage tracing experiments show the regenerate was graft derived, suggesting transformation of the chondrocytes into bone. In vitro culture data shows that cartilage explants mineralize with the addition of BMP or by exposure to HUVEC conditioned medium, indicating that endothelial cells directly promote ossification. This study provides pre-clinical data for endochondral bone repair that has potential to significantly improve patient outcomes in a variety of musculoskeletal diseases and injuries. Further, in contrast to the dogmatic view that hypertrophic chondrocytes undergo apoptosis prior to bone formation, our data suggest cartilage can transform into bone by activating the pluripotent transcription factor Oct4A. Together these data represent a paradigm shift describing the mechanism of endochondral bone repair and open the door for novel regenerative strategies based on improved biology. PMID:24259230
Cell surface reactivity of Synechococcus sp. PCC 7002: Implications for metal sorption from seawater
NASA Astrophysics Data System (ADS)
Liu, Yuxia; Alessi, D. S.; Owttrim, G. W.; Petrash, D. A.; Mloszewska, A. M.; Lalonde, S. V.; Martinez, R. E.; Zhou, Qixing; Konhauser, K. O.
2015-11-01
The past two decades have seen a significant advancement in our understanding of bacterial surface chemistry and the ability of microbes to bind metals from aqueous solutions. Much of this work has been aimed at benthic, mat-forming species in an effort to model the mechanisms by which microbes may exert control over metal contaminant transport in soils and groundwater. However, there is a distinct paucity of information pertaining to the surface chemistry of marine planktonic species, and their ability to bind trace metals from the ocean's photic zone. To this end, the surface properties of the cyanobacterium Synechococcus sp. PCC 7002 were studied as this genus is one of the dominant marine phytoplankton, and as such, contributes significantly to metal cycling in the ocean's photic zone. Zeta potential measurement indicates that the cell surfaces display a net negative charge. This was supported by potentiometric titration and Fourier transform infrared spectroscopy analyses demonstrating that the cells are dominated by surface proton releasing ligands, including carboxyl, phosphoryl and amino functional groups, with a total ligand density of 34.18 ± 1.62 mmol/g (dry biomass). Cd adsorption experiments further reveal that carboxyl groups play a primary role in metal adsorption, with 1.0 g of dry biomass binding an equivalent of 7.05 × 10-5 M of Cd from solution at pH = 8. To put this value into context, in 1 L of seawater, and with an open-ocean population of Synechococcus of 105 cells/mL in the photic zone, approximately 10 nmol of Cd could potentially be adsorbed by the cyanobacteria; an amount equivalent to seawater Cd concentrations. Although we have only focused on one microbial species and one metal cation, and we have not considered trace element assimilation, our results highlight the potential role of surface sorption by phytoplankton in the cycling of metals in the ocean.
NASA Astrophysics Data System (ADS)
Lin, Chung Ching; Lin, Po-Yen; Chang, Chia-Ching
Apolipoprotein B (apoB) is the only protein of LDL. LDL delivers cholesterol, triacylglycerides and lipids to the target cells. Reconstitute apoB lipoparticle (rABL) will be an idea drug delivery vehicle for hydrophobic and amphiphilic materials delivery. It is challenged to renature ApoB into its functional state from denatured state. By using modified bile salt and radicicol (Rad) added over-critical refolding process, apoB can be restored into its native like state. The intrinsic fluorescence of apoB increased during the refolding process. Moreover, radicicol (Rad) molecules have been encapsulated into reconstitute rABL (Rad@rABL). To investigate the cell uptake mechanism of Rad@rABL, a super resolution ground state depletion (GSD) microscopy is used in this research. Fluorescence labeled Rad@rABL can be traced within the tumor cell. Key words: LDL, radicicol, protein refolding, super resolution microscopy.
Provenance Storage, Querying, and Visualization in PBase
DOE Office of Scientific and Technical Information (OSTI.GOV)
Kianmajd, Parisa; Ludascher, Bertram; Missier, Paolo
2015-01-01
We present PBase, a repository for scientific workflows and their corresponding provenance information that facilitates the sharing of experiments among the scientific community. PBase is interoperable since it uses ProvONE, a standard provenance model for scientific workflows. Workflows and traces are stored in RDF, and with the support of SPARQL and the tree cover encoding, the repository provides a scalable infrastructure for querying the provenance data. Furthermore, through its user interface, it is possible to: visualize workflows and execution traces; visualize reachability relations within these traces; issue SPARQL queries; and visualize query results.
Pathman, Thanujeni; Bauer, Patricia J
2013-02-01
The first years of life are witness to rapid changes in long-term recall ability. In the current research we contributed to an explanation of the changes by testing the absolute and relative contributions to long-term recall of encoding and post-encoding processes. Using elicited imitation, we sampled the status of 16-, 20-, and 24-month-old infants' memory representations at various time points after experience of events. In Experiment 1, infants were tested immediately, 1 week after encoding, and again after 1 month. The measure of 1-week trace status was a unique predictor of 1-month delayed recall. In Experiment 2, infants were tested immediately, 15 min, 48 h, and 2 weeks after encoding and again 1 month later. The measures of 15-min and 48-h trace strength contributed unique variance in 1-month delayed recall. The findings highlight the need to consider post-encoding processes in explanations of variability in long-term memory during infancy. Copyright © 2012 Elsevier Inc. All rights reserved.
Amemiya, T
1999-12-01
To examine the effect of vitamins and trace elements on ocular tissue. Rats or mice were fed diets deficient in the trace elements Zn, Cu, Mn, Se, Mg, and Cr or in vitamins A, B12, C, and E. In some rats Al and vitamin A were injected in excessive amounts. We studied the conjunctiva, cornea, retina, and optic nerve with a light microscope, transmission and scanning electron microscopes, an energy dispersive X-ray analyser, and an ion microscope. Histochemical, cytochemical, and immunohistochemical techniques were applied to the pathological specimens. Deficiencies of Zn, Cu, Mn, and vitamins A, C and E caused a loss of goblet cells in the conjunctiva and a prominent decrease of microvilli and microplicae in the conjunctiva and cornea. The elements in the goblet cells were changed in these conditions. In addition, epithelial cells showed poor fibrous development and abnormal distribution of chromatin in the nucleus. Zn, Cu, Mn, and vitamins A and E deficiencies caused photoreceptor cells to degenerate and disappear. Se deficiency reduced the horizontal and amacrine cells. Vitamin B12 deficiency reduced nerve fibers in the nerve fiber layer of the retina. Mg deficiency induced multifocal necrosis in the retinal pigment epithelium and apoptotic nuclear changes in the photoreceptor cells. Cr deficiency showed abnormal phagocytosis of the photoreceptor outer segment discs in the retinal pigment epithelium. Vitamin B12 was found to be related to the circadian rhythm in the retina. Deficiencies of Zn, Cu, Mn, and vitamins A, B12, and E induced degeneration and disappearance of myelin lamellae in the myelinated optic nerve fibers. In hypervitaminosis A, lipid droplets appeared in the retinal pigment epithelium and alcohol dehydrogenase disappeared in the retinal pigment epithelium and photoreceptor outer segments. Excessive Al was toxic to the retina, which showed disappearance of photoreceptor cells. Al deposits were seen in dendrites and neurons in the outer plexiform layer. Zn seemed to be necessary for corneal epithelial cell wound healing. Trace elements usually are contained in enzymes, which have many metabolic functions. They are related to synthesis and breakdown of many substances. Some trace elements such as Zn, Cu, Mn, and Se and vitamins including vitamins A, C, and E prevent peroxidation of lipids. Some vitamins have an affinity for specific tissues such as epithelial cells, nerve fibers, and neuronal cells and are needed for cell differentiation, development, and maintenance. Cu, Zn, Mn, Se, Mg, and Cr and vitamins A, B12, C, and E are necessary for maintenance of cellular structure and metabolism.
Amemiya
2000-05-01
Purpose: To examine the effect of vitamins and trace elements on ocular tissue.Materials and Methods: Rats or mice were fed diets deficient in the trace elements Zn, Cu, Mn, Se, Mg, and Cr or in vitamins A, B(12), C, and E. In some rats Al and vitamin A were injected in excessive amounts. We studied the conjunctiva, cornea, retina, and optic nerve with a light microscope, transmission and scanning electron microscopes, an energy dispersive X-ray analyzer, and an ion microscope. Histochemical, cytochemical, and immunohistochemical techniques were applied to the pathological specimens.Results: Deficiencies of Zn, Cu, Mn, and vitamins A, C and E caused a loss of goblet cells in the conjunctiva and a prominent decrease of microvilli and microplicae in the conjunctiva and cornea. The elements in the goblet cells were changed in these conditions. In addition, epithelial cells showed poor fibrous development and abnormal distribution of chromatin in the nucleus.Zn, Cu, Mn, and vitamins A and E deficiencies caused photoreceptor cells to degenerate and disappear. Se deficiency reduced the horizontal and amacrine cells. Vitamin B(12) deficiency reduced nerve fibers in the nerve fiber layer of the retina. Mg deficiency induced multifocal necrosis in the retinal pigment epithelium and apoptotic nuclear changes in the photoreceptor cells. Cr deficiency showed abnormal phagocytosis of the photoreceptor outer segment discs in the retinal pigment epithelium. Vitamin B(12) was found to be related to the circadian rhythm in the retina.Deficiencies of Zn, Cu, Mn, and vitamins A, B(12), and E induced degeneration and disappearance of myelin lamellae in the myelinated optic nerve fibers.In hypervitaminosis A, lipid droplets appeared in the retinal pigment epithelium and alcohol dehydrogenase disappeared in the retinal pigment epithelium and photoreceptor outer segments. Excessive Al was toxic to the retina, which showed disappearance of photoreceptor cells. Al deposits were seen in dendrites and neurons in the outer plexiform layer.Zn seemed to be necessary for corneal epithelial cell wound healing.Discussion: Trace elements usually are contained in enzymes, which have many metabolic functions. They are related to synthesis and breakdown of many substances. Some trace elements such as Zn, Cu, Mn, and Se and vitamins including vitamins A, C, and E prevent peroxidation of lipids. Some vitamins have an affinity for specific tissues such as epithelial cells, nerve fibers, and neuronal cells and are needed for cell differentiation, development, and maintenance.Conclusion: Cu, Zn, Mn, Se, Mg, and Cr and vitamins A, B(12), C, and E are necessary for maintenance of cellular structure and metabolism.
Antonioletti, Mario; Biktashev, Vadim N; Jackson, Adrian; Kharche, Sanjay R; Stary, Tomas; Biktasheva, Irina V
2017-01-01
The BeatBox simulation environment combines flexible script language user interface with the robust computational tools, in order to setup cardiac electrophysiology in-silico experiments without re-coding at low-level, so that cell excitation, tissue/anatomy models, stimulation protocols may be included into a BeatBox script, and simulation run either sequentially or in parallel (MPI) without re-compilation. BeatBox is a free software written in C language to be run on a Unix-based platform. It provides the whole spectrum of multi scale tissue modelling from 0-dimensional individual cell simulation, 1-dimensional fibre, 2-dimensional sheet and 3-dimensional slab of tissue, up to anatomically realistic whole heart simulations, with run time measurements including cardiac re-entry tip/filament tracing, ECG, local/global samples of any variables, etc. BeatBox solvers, cell, and tissue/anatomy models repositories are extended via robust and flexible interfaces, thus providing an open framework for new developments in the field. In this paper we give an overview of the BeatBox current state, together with a description of the main computational methods and MPI parallelisation approaches.
Pericyte–fibroblast transition promotes tumor growth and metastasis
Hosaka, Kayoko; Yang, Yunlong; Seki, Takahiro; Fischer, Carina; Dubey, Olivier; Fredlund, Erik; Hartman, Johan; Religa, Piotr; Ishii, Yoko; Sasahara, Masakiyo; Larsson, Ola; Cossu, Giulio; Cao, Renhai; Lim, Sharon; Cao, Yihai
2016-01-01
Vascular pericytes, an important cellular component in the tumor microenvironment, are often associated with tumor vasculatures, and their functions in cancer invasion and metastasis are poorly understood. Here we show that PDGF-BB induces pericyte–fibroblast transition (PFT), which significantly contributes to tumor invasion and metastasis. Gain- and loss-of-function experiments demonstrate that PDGF-BB-PDGFRβ signaling promotes PFT both in vitro and in in vivo tumors. Genome-wide expression analysis indicates that PDGF-BB–activated pericytes acquire mesenchymal progenitor features. Pharmacological inhibition and genetic deletion of PDGFRβ ablate the PDGF-BB–induced PFT. Genetic tracing of pericytes with two independent mouse strains, TN-AP-CreERT2:R26R-tdTomato and NG2-CreERT2:R26R-tdTomato, shows that PFT cells gain stromal fibroblast and myofibroblast markers in tumors. Importantly, coimplantation of PFT cells with less-invasive tumor cells in mice markedly promotes tumor dissemination and invasion, leading to an increased number of circulating tumor cells and metastasis. Our findings reveal a mechanism of vascular pericytes in PDGF-BB–promoted cancer invasion and metastasis by inducing PFT, and thus targeting PFT may offer a new treatment option of cancer metastasis. PMID:27608497
Transition zone structure beneath Ethiopia from 3-D fast marching pseudo-migration stacking
NASA Astrophysics Data System (ADS)
Benoit, M. H.; Lopez, A.; Levin, V.
2008-12-01
Several models for the origin of the Afar hotspot have been put forth over the last decade, but much ambiguity remains as to whether the hotspot tectonism found there is due to a shallow or deeply seated feature. Additionally, there has been much debate as to whether the hotspot owes its existence to a 'classic' mantle plume feature or if it is part of the African Superplume complex. To further understand the origin of the hotspot, we employ a new receiver function stacking method that incorporates a fast-marching three- dimensional ray tracing algorithm to improve upon existing studies of the mantle transition zone structure. Using teleseismic data from the Ethiopia Broadband Seismic Experiment and the EAGLE (Ethiopia Afar Grand Lithospheric Experiment) experiment, we stack receiver functions using a three-dimensional pseudo- migration technique to examine topography on the 410 and 660 km discontinuities. Previous methods of receiver function pseudo-migration incorporated ray tracing methods that were not able to ray trace through highly complicated 3-D structure, or the ray tracing techniques only produced 3-D time perturbations associated 1-D rays in a 3-D velocity medium. These previous techniques yielded confusing and incomplete results for when applied to the exceedingly complicated mantle structure beneath Ethiopia. Indeed, comparisons of the 1-D versus 3-D ray tracing techniques show that the 1-D technique mislocated structure laterally in the mantle by over 100 km. Preliminary results using our new technique show a shallower then average 410 km discontinuity and a deeper than average 660 km discontinuity over much of the region, suggested that the hotspot has a deep seated origin.
ACE Design Study and Experiments
1976-06-01
orthophoto on off-line printer o Automatically compute contours on UNIVAC 1108 and plot on CALCOMP o Manually trace planimetry and drainage from... orthophoto * o Manually edit and trace plotted contours to obtain completed contour manuscript* - Edit errors - Add missing contour detail - Combine...stereomodels - Contours adjusted to drainage chart and spot elevations - Referring to orthophoto , rectified photos, original photos o Normal
Kiseleva, N M; Novoseletskaya, A V; Voevodina, Ye B; Kozlov, I G; Inozemtsev, A N
2012-12-01
Apart from restoration of disordered immunological parameters, tactivin and derinat exhibit a pronounced effect on the higher integrative functions of the brain. Experiments on Wistar rats have shown that these drugs accelerated conditioning of food and defense responses. New methods for quantitative evaluation of memory trace consolidation are proposed.
A Transfer Subject: Tracing Boundary-Work and Micro-Transfer in First-Year Composition
ERIC Educational Resources Information Center
Medina, Dylan London
2017-01-01
This dissertation draws on data from a quarter-long case study implementing a number of social science and humanities research methods. This study was conducted at the University of Washington in two sections of First-Year Composition and investigates the boundary-marking interactions that occur in a writing class by tracing the experiences of…
The Sensory Nature of Episodic Memory: Sensory Priming Effects Due to Memory Trace Activation
ERIC Educational Resources Information Center
Brunel, Lionel; Labeye, Elodie; Lesourd, Mathieu; Versace, Remy
2009-01-01
The aim of this study was to provide evidence that memory and perceptual processing are underpinned by the same mechanisms. Specifically, the authors conducted 3 experiments that emphasized the sensory aspect of memory traces. They examined their predictions with a short-term priming paradigm based on 2 distinct phases: a learning phase consisting…
A photoacoustic spectrometer for trace gas detection
NASA Astrophysics Data System (ADS)
Telles, E. M.; Bezerra, E.; Scalabrin, A.
2005-06-01
A high-resolution external laser photoacoustic spectrometer has been developed for trace gas detection with absorption transitions in coincidence with CO2 laser emission lines (9,2-10,9 μm: 920-1086 cm-1). The CO2 laser operates in 90 CW lines with power of up to 15 W. A PC-controlled step motor can tune the laser lines. The resonance frequency of first longitudinal mode of the photoacoustic cell is at 1600 Hz. The cell Q-factor and cell constant are measured close to 50 and 28 mVcmW-1, respectively. The spectrometer has been tested in preliminary studies to analyze the absorption transitions of ozone (O_3). The ethylene (C_2H_4) from papaya fruit is also investigated using N2 as carrier gas at a constant flow rate.
CRISPR-UMI: single-cell lineage tracing of pooled CRISPR-Cas9 screens.
Michlits, Georg; Hubmann, Maria; Wu, Szu-Hsien; Vainorius, Gintautas; Budusan, Elena; Zhuk, Sergei; Burkard, Thomas R; Novatchkova, Maria; Aichinger, Martin; Lu, Yiqing; Reece-Hoyes, John; Nitsch, Roberto; Schramek, Daniel; Hoepfner, Dominic; Elling, Ulrich
2017-12-01
Pooled CRISPR screens are a powerful tool for assessments of gene function. However, conventional analysis is based exclusively on the relative abundance of integrated single guide RNAs (sgRNAs) between populations, which does not discern distinct phenotypes and editing outcomes generated by identical sgRNAs. Here we present CRISPR-UMI, a single-cell lineage-tracing methodology for pooled screening to account for cell heterogeneity. We generated complex sgRNA libraries with unique molecular identifiers (UMIs) that allowed for screening of clonally expanded, individually tagged cells. A proof-of-principle CRISPR-UMI negative-selection screen provided increased sensitivity and robustness compared with conventional analysis by accounting for underlying cellular and editing-outcome heterogeneity and detection of outlier clones. Furthermore, a CRISPR-UMI positive-selection screen uncovered new roadblocks in reprogramming mouse embryonic fibroblasts as pluripotent stem cells, distinguishing reprogramming frequency and speed (i.e., effect size and probability). CRISPR-UMI boosts the predictive power, sensitivity, and information content of pooled CRISPR screens.
Neuronize: a tool for building realistic neuronal cell morphologies
Brito, Juan P.; Mata, Susana; Bayona, Sofia; Pastor, Luis; DeFelipe, Javier; Benavides-Piccione, Ruth
2013-01-01
This study presents a tool, Neuronize, for building realistic three-dimensional models of neuronal cells from the morphological information extracted through computer-aided tracing applications. Neuronize consists of a set of methods designed to build 3D neural meshes that approximate the cell membrane at different resolution levels, allowing a balance to be reached between the complexity and the quality of the final model. The main contribution of the present study is the proposal of a novel approach to build a realistic and accurate 3D shape of the soma from the incomplete information stored in the digitally traced neuron, which usually consists of a 2D cell body contour. This technique is based on the deformation of an initial shape driven by the position and thickness of the first order dendrites. The addition of a set of spines along the dendrites completes the model, building a final 3D neuronal cell suitable for its visualization in a wide range of 3D environments. PMID:23761740
Imaging trace element distributions in single organelles and subcellular features
Kashiv, Yoav; Austin, Jotham R.; Lai, Barry; ...
2016-02-25
The distributions of chemical elements within cells are of prime importance in a wide range of basic and applied biochemical research. An example is the role of the subcellular Zn distribution in Zn homeostasis in insulin producing pancreatic beta cells and the development of type 2 diabetes mellitus. We combined transmission electron microscopy with micro-and nano-synchrotron X-ray fluorescence to image unequivocally for the first time, to the best of our knowledge, the natural elemental distributions, including those of trace elements, in single organelles and other subcellular features. Detected elements include Cl, K, Ca, Co, Ni, Cu, Zn and Cd (whichmore » some cells were supplemented with). Cell samples were prepared by a technique that minimally affects the natural elemental concentrations and distributions, and without using fluorescent indicators. In conclusion, it could likely be applied to all cell types and provide new biochemical insights at the single organelle level not available from organelle population level studies.« less
Neuronize: a tool for building realistic neuronal cell morphologies.
Brito, Juan P; Mata, Susana; Bayona, Sofia; Pastor, Luis; Defelipe, Javier; Benavides-Piccione, Ruth
2013-01-01
This study presents a tool, Neuronize, for building realistic three-dimensional models of neuronal cells from the morphological information extracted through computer-aided tracing applications. Neuronize consists of a set of methods designed to build 3D neural meshes that approximate the cell membrane at different resolution levels, allowing a balance to be reached between the complexity and the quality of the final model. The main contribution of the present study is the proposal of a novel approach to build a realistic and accurate 3D shape of the soma from the incomplete information stored in the digitally traced neuron, which usually consists of a 2D cell body contour. This technique is based on the deformation of an initial shape driven by the position and thickness of the first order dendrites. The addition of a set of spines along the dendrites completes the model, building a final 3D neuronal cell suitable for its visualization in a wide range of 3D environments.
A wearable stimulation bandage for electrotherapy studies in a rat ischemic wound model.
Howe, Daniel S; Dunning, Jeremy L; Henzel, Mary K; Graebert, Jennifer K; Bogie, Kath M
2011-01-01
The clinical efficacy of electro-therapy in the treatment of chronic wounds is currently debated, and a in-vivo evaluation of stimulation parameters will provide the statistical evidence needed to direct clinical guidelines. A low-cost, wearable electrical stimulation bandage has been developed for use with an established rat ischemic wound model. The bandage consists of a user-programmable stimulator PCB and a plastic bandage with two hydrogel electrodes. The battery-powered bandage may be used for up to seven days between dressing changes, and the stimulator may be reused. The microcontroller-based stimulator uses a boost converter circuit to generate pulses up to 90 V from a 3 V coin cell battery. Consistent operation of the boost converter over the wide input and output voltage ranges is achieved using voltage feedforward and soft-start techniques implemented in firmware. The bandages are laser-cut to shape, and electrical traces are applied using stencils and conductive nickel paint. Both the PCB and electrical traces are encapsulated to protect the animal. The device has been successfully demonstrated using the rat ischemic wound model for a period of seven days, and clinical experiments are ongoing.
Short- and long-term memory in Drosophila require cAMP signaling in distinct neuron types.
Blum, Allison L; Li, Wanhe; Cressy, Mike; Dubnau, Josh
2009-08-25
A common feature of memory and its underlying synaptic plasticity is that each can be dissected into short-lived forms involving modification or trafficking of existing proteins and long-term forms that require new gene expression. An underlying assumption of this cellular view of memory consolidation is that these different mechanisms occur within a single neuron. At the neuroanatomical level, however, different temporal stages of memory can engage distinct neural circuits, a notion that has not been conceptually integrated with the cellular view. Here, we investigated this issue in the context of aversive Pavlovian olfactory memory in Drosophila. Previous studies have demonstrated a central role for cAMP signaling in the mushroom body (MB). The Ca(2+)-responsive adenylyl cyclase RUTABAGA is believed to be a coincidence detector in gamma neurons, one of the three principle classes of MB Kenyon cells. We were able to separately restore short-term or long-term memory to a rutabaga mutant with expression of rutabaga in different subsets of MB neurons. Our findings suggest a model in which the learning experience initiates two parallel associations: a short-lived trace in MB gamma neurons, and a long-lived trace in alpha/beta neurons.
Nokia, Miriam S; Mikkonen, Jarno E; Penttonen, Markku; Wikgren, Jan
2012-01-01
Oscillations in hippocampal local-field potentials (LFPs) reflect the crucial involvement of the hippocampus in memory trace formation: theta (4-8 Hz) oscillations and ripples (~200 Hz) occurring during sharp waves are thought to mediate encoding and consolidation, respectively. During sharp wave-ripple complexes (SPW-Rs), hippocampal cell firing closely follows the pattern that took place during the initial experience, most likely reflecting replay of that event. Disrupting hippocampal ripples using electrical stimulation either during training in awake animals or during sleep after training retards spatial learning. Here, adult rabbits were trained in trace eyeblink conditioning, a hippocampus-dependent associative learning task. A bright light was presented to the animals during the inter-trial interval (ITI), when awake, either during SPW-Rs or irrespective of their neural state. Learning was particularly poor when the light was presented following SPW-Rs. While the light did not disrupt the ripple itself, it elicited a theta-band oscillation, a state that does not usually coincide with SPW-Rs. Thus, it seems that consolidation depends on neuronal activity within and beyond the hippocampus taking place immediately after, but by no means limited to, hippocampal SPW-Rs.
Moving epithelia: Tracking the fate of mammalian limbal epithelial stem cells.
Di Girolamo, Nick
2015-09-01
Lineage tracing allows the destiny of a stem cell (SC) and its progeny to be followed through time. In order to track their long-term fate, SC must be permanently marked to discern their distribution, division, displacement and differentiation. This information is essential for unravelling the mysteries that govern their replenishing activity while they remain anchored within their niche microenvironment. Modern-day lineage tracing uses inducible genetic recombination to illuminate cells within embryonic, newborn and adult tissues, and the advent of powerful high-resolution microscopy has enabled the behaviour of labelled cells to be monitored in real-time in a living organism. The simple structural organization of the mammalian cornea, including its accessibility and transparency, renders it the ideal tissue to study SC fate using lineage tracing assisted by non-invasive intravital microscopy. Despite more than a century of research devoted to understanding how this tissue is maintained and repaired, many limitations and controversies continue to plague the field, including uncertainties about the specificity of current SC markers, the number of SC within the cornea, their mode of division, their location, and importantly the signals that dictate cell migration. This communication will highlight historical discoveries as well as recent developments in the corneal SC field; more specifically how the progeny of these cells are mobilised to replenish this dynamic tissue during steady-state, disease and transplantation. Also discussed is how insights gleaned from animal studies can be used to advance our knowledge of the fundamental mechanisms that govern modelling and remodelling of the human cornea in health and disease. Copyright © 2015 Elsevier Ltd. All rights reserved.
Nakajima, Kazuki; Ito, Emi; Ohtsubo, Kazuaki; Shirato, Ken; Takamiya, Rina; Kitazume, Shinobu; Angata, Takashi; Taniguchi, Naoyuki
2013-01-01
Nucleotide sugars are the donor substrates of various glycosyltransferases, and an important building block in N- and O-glycan biosynthesis. Their intercellular concentrations are regulated by cellular metabolic states including diseases such as cancer and diabetes. To investigate the fate of UDP-GlcNAc, we developed a tracing method for UDP-GlcNAc synthesis and use, and GlcNAc utilization using 13C6-glucose and 13C2-glucosamine, respectively, followed by the analysis of mass isotopomers using LC-MS. Metabolic labeling of cultured cells with 13C6-glucose and the analysis of isotopomers of UDP-HexNAc (UDP-GlcNAc plus UDP-GalNAc) and CMP-NeuAc revealed the relative contributions of metabolic pathways leading to UDP-GlcNAc synthesis and use. In pancreatic insulinoma cells, the labeling efficiency of a 13C6-glucose motif in CMP-NeuAc was lower compared with that in hepatoma cells. Using 13C2-glucosamine, the diversity of the labeling efficiency was observed in each sugar residue of N- and O-glycans on the basis of isotopomer analysis. In the insulinoma cells, the low labeling efficiencies were found for sialic acids as well as tri- and tetra-sialo N-glycans, whereas asialo N-glycans were found to be abundant. Essentially no significant difference in secreted hyaluronic acids was found among hepatoma and insulinoma cell lines. This indicates that metabolic flows are responsible for the low sialylation in the insulinoma cells. Our strategy should be useful for systematically tracing each stage of cellular GlcNAc metabolism. PMID:23720760
Corbin, Jonathan C.; Reyna, Valerie F.; Weldon, Rebecca B.; Brainerd, Charles J.
2015-01-01
Fuzzy-trace theory distinguishes verbatim (literal, exact) from gist (meaningful) representations, predicting that reliance on gist increases with experience and expertise. Thus, many judgment-and-decision-making biases increase with development, such that cognition is colored by context in ways that violate logical coherence and probability theories. Nevertheless, this increase in gist-based intuition is adaptive: Gist is stable, less sensitive to interference, and easier to manipulate. Moreover, gist captures the functionally significant essence of information, supporting healthier and more robust decision processes. We describe how fuzzy-trace theory accounts for judgment-and-decision making phenomena, predicting the paradoxical arc of these processes with the development of experience and expertise. We present data linking gist memory processes to gist processing in decision making and provide illustrations of gist reliance in medicine, public health, and intelligence analysis. PMID:26664820
Corbin, Jonathan C; Reyna, Valerie F; Weldon, Rebecca B; Brainerd, Charles J
2015-12-01
Fuzzy-trace theory distinguishes verbatim (literal, exact) from gist (meaningful) representations, predicting that reliance on gist increases with experience and expertise. Thus, many judgment-and-decision-making biases increase with development, such that cognition is colored by context in ways that violate logical coherence and probability theories. Nevertheless, this increase in gist-based intuition is adaptive: Gist is stable, less sensitive to interference, and easier to manipulate. Moreover, gist captures the functionally significant essence of information, supporting healthier and more robust decision processes. We describe how fuzzy-trace theory accounts for judgment-and-decision making phenomena, predicting the paradoxical arc of these processes with the development of experience and expertise. We present data linking gist memory processes to gist processing in decision making and provide illustrations of gist reliance in medicine, public health, and intelligence analysis.
Verma, Sunita; Worden, John; Payra, Swagata; Jourdain, Line; Shim, Changsub
2009-07-01
A major aircraft experiment Transport and Chemical Evolution over the Pacific (TRACE-P) mission over the NW Pacific in March-April 2001 was conducted to better understand how outflow from the Asian continent affects the composition of the global atmosphere. In this paper, a global climate model, GEOS-Chem is used to investigate possible black carbon aerosol contributions from TRACE-P region. Our result depicts that absorbing black carbon ("soot") significantly outflow during lifting to the free troposphere through warm conveyor belt and convection associated with this lifting. The GEOS-Chem simulation results show significant transport of black carbon aerosols from Asian regions to the Western Pacific region during the spring season. As estimated by GEOS-Chem simulations, approximately 25% of the black carbon concentrations over the western pacific originate from SE Asia in the spring.
Deonarine, Amrika; Kolker, Allan; Doughten, Michael W.
2015-01-01
In this fact sheet, the form, distribution, and behavior of trace elements of environmental interest in samples of coal fly ash were investigated in response to concerns about element mobility in the event of an ash spill. The study includes laboratory-based leaching experiments to examine the behavior of trace elements, such as arsenic (As) and chromium (Cr), in response to key environmental factors including redox conditions (degree of oxygenation), which are known to vary with depth within coal ash impoundments and in natural ecosystems. The experiments show that As dissolves from samples of coal fly ash into simulated freshwater under both oxic (highly oxygenated) and anoxic (poorly oxygenated) conditions, whereas dissolved Cr concentrations are very redox dependent. This U.S. Geological Survey research helps define the distribution of elements such as As in coal ash and shows that element mobility can vary considerably under different conditions expected in the environment.
Dr. Garry Latham - Seismometer Tracings Study - Moon - MSC
1969-07-22
S69-39588 (20 July 1969) --- Dr. Garry Latham, with the Lamont Geological Observatory, studies seismometer tracings in the Mission Control Center?s ASEP control room. The electronic data was coming from the Passive Seismic Experiments Package which the Apollo 11 astronauts had just deployed on the surface of the moon. Dr. Lamont is the principal investigator for the PSEP, a component of the Early Apollo Scientific Experiments Package (EASEP). PSEP uses three long-period seismometers and one short-period vertical seismometer for measuring meteoroid impacts and moonquakes. Such data will be useful in determining the interior structure of the moon; for example, does the moon have a core and mantle like Earth? Here, the center trace shows evidence of activity on the moon. The PSEP was sensitive enough to pick up the footsteps of astronauts Neil A. Armstrong and Edwin E. Aldrin Jr. as they walked on the moon.
Dr. Garry Latham studies seismometer tracings from the moon
1969-07-22
S69-39587 (20 July 1969) --- Dr. Garry Latham (left) with the Lamont Geological Observatory, studies seismometer tracings in the Mission Control Center's (MCC) ALSEP control room. The electronic data was coming from the Passive Seismic Experiments Package (PSEP) which the Apollo 11 astronauts had just deployed on the surface of the moon. Dr. Lamont is the principal investigator for the PSEP, a component of the Early Apollo Scientific Experiments Package (EASEP). PSEP uses three long-period seismometers and one short-period vertical seismometer for measuring meteoroid impacts and moonquakes. Such data will be useful in determining the interior structure of the moon; for example, does the moon have a core and mantle like Earth? Here, the flapping of the PSEP's solar panels is picked up and registered as a tracing. The PSEP was sensitive enough to pick up the footsteps of astronauts Neil A. Armstrong and Edwin E. Aldrin Jr., as they walked on the moon.
Pan, Yunyu; Koopmans, Gerwin F; Bonten, Luc T C; Song, Jing; Luo, Yongming; Temminghoff, Erwin J M; Comans, Rob N J
2016-12-01
Alternating flooding and drainage conditions have a strong influence on redox chemistry and the solubility of trace metals in paddy soils. However, current knowledge of how the effects of water management on trace metal solubility are linked to trace metal uptake by rice plants over time is still limited. Here, a field-contaminated paddy soil was subjected to two flooding and drainage cycles in a pot experiment with two rice plant cultivars, exhibiting either high or low Cd accumulation characteristics. Flooding led to a strong vertical gradient in the redox potential (Eh). The pH and Mn, Fe, and dissolved organic carbon concentrations increased with decreasing Eh and vice versa. During flooding, trace metal solubility decreased markedly, probably due to sulfide mineral precipitation. Despite its low solubility, the Cd content in rice grains exceeded the food quality standards for both cultivars. Trace metal contents in different rice plant tissues (roots, stem, and leaves) increased at a constant rate during the first flooding and drainage cycle but decreased after reaching a maximum during the second cycle. As such, the high temporal variability in trace metal solubility was not reflected in trace metal uptake by rice plants over time. This might be due to the presence of aerobic conditions and a consequent higher trace metal solubility near the root surface, even during flooding. Trace metal solubility in the rhizosphere should be considered when linking water management to trace metal uptake by rice over time.
Generation of enteroendocrine cell diversity in midgut stem cell lineages
Beehler-Evans, Ryan; Micchelli, Craig A.
2015-01-01
The endocrine system mediates long-range peptide hormone signaling to broadcast changes in metabolic status to distant target tissues via the circulatory system. In many animals, the diffuse endocrine system of the gut is the largest endocrine tissue, with the full spectrum of endocrine cell subtypes not yet fully characterized. Here, we combine molecular mapping, lineage tracing and genetic analysis in the adult fruit fly to gain new insight into the cellular and molecular mechanisms governing enteroendocrine cell diversity. Neuropeptide hormone distribution was used as a basis to generate a high-resolution cellular map of the diffuse endocrine system. Our studies show that cell diversity is seen at two distinct levels: regional and local. We find that class I and class II enteroendocrine cells can be distinguished locally by combinatorial expression of secreted neuropeptide hormones. Cell lineage tracing studies demonstrate that class I and class II cells arise from a common stem cell lineage and that peptide profiles are a stable feature of enteroendocrine cell identity during homeostasis and following challenge with the enteric pathogen Pseudomonas entomophila. Genetic analysis shows that Notch signaling controls the establishment of class II cells in the lineage, but is insufficient to reprogram extant class I cells into class II enteroendocrine cells. Thus, one mechanism by which secretory cell diversity is achieved in the diffuse endocrine system is through cell-cell signaling interactions within individual adult stem cell lineages. PMID:25670792
Schwartz, Andrea G; Long, Fanxin; Thomopoulos, Stavros
2015-01-01
Tendon attaches to bone across a specialized tissue called the enthesis. This tissue modulates the transfer of muscle forces between two materials, i.e. tendon and bone, with vastly different mechanical properties. The enthesis for many tendons consists of a mineralized graded fibrocartilage that develops postnatally, concurrent with epiphyseal mineralization. Although it is well described that the mineralization and development of functional maturity requires muscle loading, the biological factors that modulate enthesis development are poorly understood. By genetically demarcating cells expressing Gli1 in response to Hedgehog (Hh) signaling, we discovered a unique population of Hh-responsive cells in the developing murine enthesis that were distinct from tendon fibroblasts and epiphyseal chondrocytes. Lineage-tracing experiments revealed that the Gli1 lineage cells that originate in utero eventually populate the entire mature enthesis. Muscle paralysis increased the number of Hh-responsive cells in the enthesis, demonstrating that responsiveness to Hh is modulated in part by muscle loading. Ablation of the Hh-responsive cells during the first week of postnatal development resulted in a loss of mineralized fibrocartilage, with very little tissue remodeling 5 weeks after cell ablation. Conditional deletion of smoothened, a molecule necessary for responsiveness to Ihh, from the developing tendon and enthesis altered the differentiation of enthesis progenitor cells, resulting in significantly reduced fibrocartilage mineralization and decreased biomechanical function. Taken together, these results demonstrate that Hh signaling within developing enthesis fibrocartilage cells is required for enthesis formation. © 2015. Published by The Company of Biologists Ltd.
Schwartz, Andrea G.; Long, Fanxin; Thomopoulos, Stavros
2015-01-01
Tendon attaches to bone across a specialized tissue called the enthesis. This tissue modulates the transfer of muscle forces between two materials, i.e. tendon and bone, with vastly different mechanical properties. The enthesis for many tendons consists of a mineralized graded fibrocartilage that develops postnatally, concurrent with epiphyseal mineralization. Although it is well described that the mineralization and development of functional maturity requires muscle loading, the biological factors that modulate enthesis development are poorly understood. By genetically demarcating cells expressing Gli1 in response to Hedgehog (Hh) signaling, we discovered a unique population of Hh-responsive cells in the developing murine enthesis that were distinct from tendon fibroblasts and epiphyseal chondrocytes. Lineage-tracing experiments revealed that the Gli1 lineage cells that originate in utero eventually populate the entire mature enthesis. Muscle paralysis increased the number of Hh-responsive cells in the enthesis, demonstrating that responsiveness to Hh is modulated in part by muscle loading. Ablation of the Hh-responsive cells during the first week of postnatal development resulted in a loss of mineralized fibrocartilage, with very little tissue remodeling 5 weeks after cell ablation. Conditional deletion of smoothened, a molecule necessary for responsiveness to Ihh, from the developing tendon and enthesis altered the differentiation of enthesis progenitor cells, resulting in significantly reduced fibrocartilage mineralization and decreased biomechanical function. Taken together, these results demonstrate that Hh signaling within developing enthesis fibrocartilage cells is required for enthesis formation. PMID:25516975
Katherine Philpott, M; Stanciu, Cristina E; Kwon, Ye Jin; Bustamante, Eduardo E; Greenspoon, Susan A; Ehrhardt, Christopher J
2017-07-01
The goal of this study was to survey optical and biochemical variation in cell populations deposited onto a surface through touch or contact and identify specific features that may be used to distinguish and then sort cell populations from separate contributors in a trace biological mixture. Although we were not able to detect meaningful biochemical variation in touch samples deposited by different contributors through preliminary antibody surveys, we did observe distinct differences in red autofluorescence emissions (650-670 nm), with as much as a tenfold difference in mean fluorescence intensities observed between certain pairs of donors. Results indicate that the level of red autofluorescence in touch samples can be influenced by a donor's contact with specific material prior to handling the substrate from which cells were collected. In particular, we observed increased red autofluorescence in cells deposited subsequent to handling laboratory gloves, plant material, and certain types of marker ink, which could be easily visualized microscopically or using flow cytometry, and persisted after hand washing. To test whether these observed optical differences could potentially be used as the basis for a cell separation workflow, a controlled two-person touch mixture was separated into two fractions via fluorescence-activated cell sorting (FACS) using gating criteria based on intensity of 650-670 nm emissions and then subjected to DNA analysis. Genetic analysis of the sorted fractions provided partial DNA profiles that were consistent with separation of individual contributors from the mixture suggesting that variation in autofluorescence signatures, even if driven by extrinsic factors, may nonetheless be a useful means of isolating contributors to some touch mixtures. Graphical Abstract Conceptual workflow diagram. Trace biological mixtures containing cells from multiple individuals are analyzed by flow cytometry. Cells are then physically separated into two populations based on intensity of red autofluorescence using Fluorescence Activated Cell Sorting. Each isolated cell fraction is subjected to DNA analysis resulting in a DNA profile for each contributor.
Single Cell-Based Vector Tracing in Patients with ADA-SCID Treated with Stem Cell Gene Therapy.
Igarashi, Yuka; Uchiyama, Toru; Minegishi, Tomoko; Takahashi, Sirirat; Watanabe, Nobuyuki; Kawai, Toshinao; Yamada, Masafumi; Ariga, Tadashi; Onodera, Masafumi
2017-09-15
Clinical improvement in stem cell gene therapy (SCGT) for primary immunodeficiencies depends on the engraftment levels of genetically corrected cells, and tracing the transgene in each hematopoietic lineage is therefore extremely important in evaluating the efficacy of SCGT. We established a single cell-based droplet digital PCR (sc-ddPCR) method consisting of the encapsulation of a single cell into each droplet, followed by emulsion PCR with primers and probes specific for the transgene. A fluorescent signal in a droplet indicates the presence of a single cell carrying the target gene in its genome, and this system can clearly determine the ratio of transgene-positive cells in the entire population at the genomic level. Using sc-ddPCR, we analyzed the engraftment of vector-transduced cells in two patients with severe combined immunodeficiency (SCID) who were treated with SCGT. Sufficient engraftment of the transduced cells was limited to the T cell lineage in peripheral blood (PB), and a small percentage of CD34 + cells exhibited vector integration in bone marrow, indicating that the transgene-positive cells in PB might have differentiated from a small population of stem cells or lineage-restricted precursor cells. sc-ddPCR is a simplified and powerful tool for the detailed assessment of transgene-positive cell distribution in patients treated with SCGT.
Patel, Jatin; Seppanen, Elke J; Rodero, Mathieu P; Wong, Ho Yi; Donovan, Prudence; Neufeld, Zoltan; Fisk, Nicholas M; Francois, Mathias; Khosrotehrani, Kiarash
2017-02-21
During adult life, blood vessel formation is thought to occur via angiogenic processes involving branching from existing vessels. An alternate proposal suggests that neovessels form from endothelial progenitors able to assemble the intimal layers. We here aimed to define vessel-resident endothelial progenitors in vivo in a variety of tissues in physiological and pathological situations such as normal aorta, lungs, and wound healing, tumors, and placenta, as well. Based on protein expression levels of common endothelial markers using flow cytometry, 3 subpopulations of endothelial cells could be identified among VE-Cadherin+ and CD45- cells. Lineage tracing by using Cdh5cre ERt2 /Rosa-YFP reporter strategy demonstrated that the CD31-/loVEGFR2lo/intracellular endothelial population was indeed an endovascular progenitor (EVP) of an intermediate CD31intVEGFR2lo/intracellular transit amplifying (TA) and a definitive differentiated (D) CD31hiVEGFR2hi/extracellular population. EVP cells arose from vascular-resident beds that could not be transferred by bone marrow transplantation. Furthermore, EVP displayed progenitor-like status with a high proportion of cells in a quiescent cell cycle phase as assessed in wounds, tumors, and aorta. Only EVP cells and not TA and D cells had self-renewal capacity as demonstrated by colony-forming capacity in limiting dilution and by transplantation in Matrigel plugs in recipient mice. RNA sequencing revealed prominent gene expression differences between EVP and D cells. In particular, EVP cells highly expressed genes related to progenitor function including Sox9 , Il33 , Egfr , and Pdfgrα. Conversely, D cells highly expressed genes related to differentiated endothelium including Ets1&2 , Gata2 , Cd31 , Vwf , and Notch . The RNA sequencing also pointed to an essential role of the Sox18 transcription factor. The role of SOX18 in the differentiation process was validated by using lineage-tracing experiments based on S ox18Cre ERt2 /Rosa-YFP mice. Besides, in the absence of functional SOX18/SOXF, EVP progenitors were still present, but TA and D populations were significantly reduced. Our findings support an entirely novel endothelial hierarchy, from EVP to TA to D, as defined by self-renewal, differentiation, and molecular profiling of an endothelial progenitor. This paradigm shift in our understanding of vascular-resident endothelial progenitors in tissue regeneration opens new avenues for better understanding of cardiovascular biology. © 2016 American Heart Association, Inc.
Belostotsky, Inessa; Gridin, Vladimir V; Schechter, Israel; Yarnitzky, Chaim N
2003-02-01
An improved analytical method for airborne lead traces is reported. It is based on using a Venturi scrubber sampling device for simultaneous thin-film stripping and droplet entrapment of aerosol influxes. At least threefold enhancement of the lead-trace pre-concentration is achieved. The sampled traces are analyzed by square-wave anodic stripping voltammetry. The method was tested by a series of pilot experiments. These were performed using contaminant-controlled air intakes. Reproducible calibration plots were obtained. The data were validated by traditional analysis using filter sampling. LODs are comparable with the conventional techniques. The method was successfully applied to on-line and in situ environmental monitoring of lead.
Medvedev, Nickolay S; Shaverina, Anastasiya V; Tsygankova, Alphiya R; Saprykin, Anatoly I
2016-08-01
The paper presents a combined technique of germanium dioxide analysis by inductively coupled plasma atomic emission spectrometry (ICP-AES) with preconcentration of trace elements by distilling off matrix and electrothermal (ETV) introduction of the trace elements concentrate into the ICP. Evaluation of metrological characteristics of the developed technique of high-purity germanium dioxide analysis was performed. The limits of detection (LODs) for 25 trace elements ranged from 0.05 to 20ng/g. The accuracy of proposed technique is confirmed by "added-found" («or spiking») experiment and comparing the results of ETV-ICP-AES and ICP-AES analysis of high purity germanium dioxide samples. Copyright © 2016 Elsevier B.V. All rights reserved.
[Remote sensing of atmospheric trace gas by airborne passive FTIR].
Gao, Min-quang; Liu, Wen-qing; Zhang, Tian-shu; Liu, Jian-guo; Lu, Yi-huai; Wang, Ya-ping; Xu, Liang; Zhu, Jun; Chen, Jun
2006-12-01
The present article describes the details of aviatic measurement for remote sensing trace gases in atmosphere under various surface backgrounds with airborne passive FTIR. The passive down viewing and remote sensing technique used in the experiment is discussed. The method of acquiring atmospheric trace gases infrared characteristic spectra in complicated background and the algorithm of concentration retrieval are discussed. The concentrations of CO and N2O of boundary-layer atmosphere in experimental region below 1000 m are analyzed quantitatively. This measurement technique and the data analysis method, which does not require a previously measured background spectrum, allow fast and mobile remote detection and identification of atmosphere trace gas in large area, and also can be used for urgent monitoring of pollution accidental breakout.
Chandra, Rachna; Prusty, B Anjan Kumar; Azeez, P A
2014-06-01
Trace metals in soils may be inherited from the parent materials or added to the system due to anthropogenic activities. In proposed mining areas, trace metals become an integral part of the soil system. Usually, researchers undertake experiments on plant species selection (for the restoration plan) only after the termination of mining activities, i.e. without any pre-mining information about the soil-plant interactions. Though not shown in studies, it is clear that several recovery plans remain unsuccessful while carrying out restoration experiments. Therefore, we hypothesize that to restore the area effectively, it is imperative to consider the pre-mining scenario of metal levels in parent material as well as the vegetation ecology of the region. With these specifics, we examined the concentrations of trace metals in parent soils at three proposed bauxite locations in the Eastern Ghats, India, and compared them at a spatio-temporal scale. Vegetation quantification and other basic soil parameters accounted for establishing the connection between soil and plants. The study recorded significant spatial heterogeneity in trace metal concentrations and the role of vegetation on metal availability. Oxidation reduction potential (ORP), pH and cation exchange capacity (CEC) directly influenced metal content, and Cu and Ni were lithogenic in origin. It implies that for effective restoration plant species varies for each geological location.
Stratospheric H2O and HNO3 profiles derived from solar occultation measurements
NASA Technical Reports Server (NTRS)
Fischer, H.; Fergg, F.; Rabus, D.; Burkert, P.
1985-01-01
Compact two-channel radiometers for solar occultation experiments have been constructed in order to measure stratospheric trace gases. The instruments can be used as filter- or correlation-type radiometers, depending on the trace gas under investigation. Within the LIMS correlative measurement program, balloon flights were performed with a payload of up to four of these two-channel radiometers. From the filter-type measurements, profiles of the trace gases H2O and HNO3 are inferred for the height region between the tropopause and the balloon float level. The data evaluation also includes a comprehensive analysis of the error sources and their effect on the accuracy of the trace gas profiles. The derived H2O and HNO3 profiles are assessed against the observations of other authors and are discussed in the light of the trace gas distributions calcualted from photochemical models.
Testing Linear Temporal Logic Formulae on Finite Execution Traces
NASA Technical Reports Server (NTRS)
Havelund, Klaus; Rosu, Grigore; Norvig, Peter (Technical Monitor)
2001-01-01
We present an algorithm for efficiently testing Linear Temporal Logic (LTL) formulae on finite execution traces. The standard models of LTL are infinite traces, reflecting the behavior of reactive and concurrent systems which conceptually may be continuously alive. In most past applications of LTL. theorem provers and model checkers have been used to formally prove that down-scaled models satisfy such LTL specifications. Our goal is instead to use LTL for up-scaled testing of real software applications. Such tests correspond to analyzing the conformance of finite traces against LTL formulae. We first describe what it means for a finite trace to satisfy an LTL property. We then suggest an optimized algorithm based on transforming LTL formulae. The work is done using the Maude rewriting system. which turns out to provide a perfect notation and an efficient rewriting engine for performing these experiments.
[Temporal changes in conscious and unconscious trace phenomena in six to seven-year-old children].
Fedotchev, A I
1975-01-01
Development in time of unconscious trace reactions as well as a change in the time of reproduction of verbal material in natural conditions was studied by the conditioned reflex to time method on 107 school children aged from six to seven years and on 35 subjects of the same age from a kindergarten. It has been found that in schoolchildren the transfer of trace reactions to long-term storage is completed faster, and their voluntary acts in the course of conscious reproduction of an interstimuli intervals are more adequate than in the kindergarten children of the same age. The trace phenomena formed in natural conditions and in the laboratory experiment develop in time in a similar way. The change in time of conscious trace phenomena is of a more complex nature than of the unconscious phenomena. The practical significance of the data obtained is noted.
Outlines of a multiple trace theory of temporal preparation.
Los, Sander A; Kruijne, Wouter; Meeter, Martijn
2014-01-01
We outline a new multiple trace theory of temporal preparation (MTP), which accounts for behavior in reaction time (RT) tasks in which the participant is presented with a warning stimulus (S1) followed by a target stimulus (S2) that requires a speeded response. The theory assumes that during the foreperiod (FP; the S1-S2 interval) inhibition is applied to prevent premature response, while a wave of activation occurs upon the presentation of S2. On each trial, these actions are stored in a separate memory trace, which, jointly with earlier formed memory traces, starts contributing to preparation on subsequent trials. We show that MTP accounts for classic effects in temporal preparation, including mean RT-FP functions observed under a variety of FP distributions and asymmetric sequential effects. We discuss the advantages of MTP over other accounts of these effects (trace-conditioning and hazard-based explanations) and suggest a critical experiment to empirically distinguish among them.
dos Santos Mota, Ananda; Morais Monteiro, Priscila; Carvalho, Angela Cristina Gouvêa; Fernandes Diniz, Barbara; Gemal Lanzieri, Pedro; Carneiro Ramos, Ricardo; Mocarzel, Luis Otavio
2017-01-01
Diarrhea is one of the most common symptoms in common variable immunodeficiency, but neurologic manifestations are rare. We presented a 50-year-old woman with recurrent diarrhea and severe weight loss that developed a posterior cord syndrome. Endoscopy found a duodenal villous blunting, intraepithelial lymphocytosis, and lack of plasma cells and magnetic resonance imaging of the spine was normal. Laboratory assays confirmed common variable immunodeficiency syndrome and showed low levels of trace elements (copper and zinc). Treatment was initiated with parenteral replacement of trace elements and intravenous human immunoglobulin and the patient improved clinically. In conclusion, physicians must be aware that gastrointestinal and neurologic disorders may be related to each other and remember to request trace elements laboratory assessment. PMID:28356913
Cosmic Ray Studies on Skies and on Campus.
ERIC Educational Resources Information Center
Jones, Brian
1993-01-01
Outlines the highlights of experiments that allow students to trace the historical development of our understandings of cosmic rays. The experiments provide for two outdoor fieldwork experiences, indoor laboratory work, and an opportunity for a group of students to show originality and initiative. (ZWH)
Omics markers of the red cell storage lesion and metabolic linkage
D’Alessandro, Angelo; Nemkov, Travis; Reisz, Julie; Dzieciatkowska, Monika; Wither, Matthew J.; Hansen, Kirk C.
2017-01-01
The introduction of omics technologies in the field of Transfusion Medicine has significantly advanced our understanding of the red cell storage lesion. While the clinical relevance of such a lesion is still a matter of debate, quantitative and redox proteomics approaches, as well quantitative metabolic flux analysis and metabolic tracing experiments promise to revolutionise our understanding of the role of blood processing strategies, inform the design and testing of novel additives or technologies (such as pathogen reduction), and evaluate the clinical relevance of donor and recipient biological variability with respect to red cell storability and transfusion outcomes. By reviewing existing literature in this rapidly expanding research endeavour, we highlight for the first time a correlation between metabolic markers of the red cell storage age and protein markers of haemolysis. Finally, we introduce the concept of metabolic linkage, i.e. the appreciation of a network of highly correlated small molecule metabolites which results from biochemical constraints of erythrocyte metabolic enzyme activities. For the foreseeable future, red cell studies will advance Transfusion Medicine and haematology by addressing the alteration of metabolic linkage phenotypes in response to stimuli, including, but not limited to, storage additives, enzymopathies (e.g. glucose 6-phosphate dehydrogenase deficiency), hypoxia, sepsis or haemorrhage. PMID:28263171
Sub-micron Hard X-ray Fluorescence Imaging of Synthetic Elements
Jensen, Mark P.; Aryal, Baikuntha P.; Gorman-Lewis, Drew; Paunesku, Tatjana; Lai, Barry; Vogt, Stefan; Woloschak, Gayle E.
2013-01-01
Synchrotron-based X-ray fluorescence microscopy (SXFM) using hard X-rays focused into sub-micron spots is a powerful technique for elemental quantification and mapping, as well as microspectroscopic measurement such as μ-XANES (X-ray absorption near edge structure). We have used SXFM to image and simultaneously quantify the transuranic element plutonium at the L3 or L2 edge as well as lighter biologically essential elements in individual rat pheochromocytoma (PC12) cells after exposure to the long-lived plutonium isotope 242Pu. Elemental maps reveal that plutonium localizes principally in the cytoplasm of the cells and avoids the cell nucleus, which is marked by the highest concentrations of phosphorus and zinc, under the conditions of our experiments. The minimum detection limit under typical acquisition conditions for an average 202 μm2 cell is 1.4 fg Pu/cell or 2.9 × 10−20 moles Pu/μm2, which is similar to the detection limit of K-edge SXFM of transition metals at 10 keV. Copper electron microscopy grids were used to avoid interference from gold X-ray emissions, but traces of strontium present in naturally occurring calcium can still interfere with plutonium detection using its Lα X-ray emission. PMID:22444530
Chatelain, M; Gasparini, J; Frantz, A
2016-04-01
Understanding the effects of trace metals emitted by anthropogenic activities on wildlife is of great concern in urban ecology; yet, information on how they affect individuals, populations, communities and ecosystems remains scarce. In particular, trace metals may impact survival by altering the immune system response to parasites. Plumage melanin is assumed to influence the effects of trace metals on immunity owing to its ability to bind metal ions in feathers and its synthesis being coded by a pleiotropic gene. We thus hypothesized that trace metal exposure would interact with plumage colouration in shaping immune response. We experimentally investigated the interactive effect between exposure to an environmentally relevant range of zinc and/or lead and melanin-based plumage colouration on components of the immune system in feral pigeons (Columba livia). We found that zinc increased anti-keyhole limpet hemocyanin (KLH) IgY primary response maintenance, buffered the negative effect of lead on anti-KLH IgY secondary response maintenance and tended to increase T-cell mediated phytohaemagglutinin (PHA) skin response. Lead decreased the peak of the anti-KLH IgY secondary response. In addition, pheomelanic pigeons exhibited a higher secondary anti-KLH IgY response than did eumelanic ones. Finally, T-cell mediated PHA skin response decreased with increasing plumage eumelanin level of birds exposed to lead. Neither treatments nor plumage colouration correlated with endoparasite intensity. Overall, our study points out the effects of trace metals on some parameters of birds' immunity, independently from other confounding urbanization factors, and underlines the need to investigate their impacts on other life history traits and their consequences in the ecology and evolution of host-parasite interactions.
Cell Phone Roulette and "Consumer Interactive" Quality
ERIC Educational Resources Information Center
Navarro, Peter
2005-01-01
Under current policies, cell phone consumers face a lower probability of finding the best carrier for their usage patterns than winning at roulette. Corroborating survey data consistently show significant dissatisfaction among cell phone users, network performance is a major issue, and customer "churn" is high. This problem may be traced to a new…
Zhang, Li-fang; Wang, Fei; Yu, Li-bin; Yan, Jian-hua; Cen, Ke-fa
2015-06-01
In order to on-line measure the trace ammonia slip of the commercial power plant in the future, this research seeks to measure the trace ammonia by using tunable diode laser absorption spectroscopy under ambient temperature and pressure, and at different temperatures, and the measuring temperature is about 650 K in the power plant. In recent years lasers have become commercially available in the near-infrared where the transitions are much stronger, and ammonia's spectroscopy is pretty complicated and the overlapping lines are difficult to resolve. A group of ammonia transitions near 4 433.5 cm(-1) in the v2 +v3 combination band have been thoroughly selected for detecting lower concentration by analyzing its absorption characteristic and considering other absorption interference in combustion gases where H2O and CO2 mole fraction are very large. To illustrate the potential for NH3 concentration measurements, predictions for NH3, H2O and CO2 are simultaneously simulated, NH3 absorption lines near 4 433.5 cm(-1) wavelength meet weaker H2O absorption than the commercial NH3 lines, and there is almost no CO2 absorption, all the parameters are based on the HITRAN database, and an improved detection limit was obtained for interference-free NH3 monitoring, this 2.25 μm band has line strengths several times larger than absorption lines in the 1.53 μm band which was often used by NH3 sensors for emission monitoring and analyzing. The measurement system was developed with a new Herriott cell and a heated gas cell realizing fast absorption measurements of high resolution, and combined with direct absorption and wavelenguh modulation based on tunable diode laser absorption spectroscopy at different temperatures. The lorentzian line shape is dominant at ambient temperature and pressure, and the estimated detectivity is approximately 0.225 x 10(-6) (SNR = 1) for the directed absorption spectroscopy, assuming a noise-equivalent absorbance of 1 x 10(-4). The heated cell experiments with controlled the temperature were performed to validate the sensing strategy. Here the Wavelength Modulation Spectroscopy (WMS) strategy was usually used to measure lower gas concentration for high noise immunity to the non-absorption transmission losses. The great agreement 2f signal with the calibrated concentration is within the uncertainty at different temperatures by using simple digital signal processing such as multiple averages, wavelet analysis and so on. The denoise processing has a great advantage in application and implementation over other noise suppression techniques. The result provided a good basis for trace ammonia escape detection based on tunable diode laser absorption spectroscopy.
NASA Astrophysics Data System (ADS)
UŻarowska, E.; Czajkowski, Rafał; Konopka, W.
2014-11-01
We aim to create a set of genetic tools where permanent opsin expression (ChR or NpHR) is precisely limited to the population of neurons that express immediate early gene c-fos during a specific temporal window of behavioral training. Since the c-fos gene is only expressed in neurons that form experience-dependent ensemble, this approach will result in specific labeling of a small subset of cells that create memory trace for the learned behavior. To this end we employ two alternative inducible gene expression systems: Tet Expression System and Cre/lox System. In both cases, the temporal window for opsin induction is controlled pharmacologically, by doxycycline or tamoxifen, respectively. Both systems will be used for creating lines of transgenic animals.
Space shuttle electrical power generation and reactant supply system
NASA Technical Reports Server (NTRS)
Simon, W. E.
1985-01-01
The design philosophy and development experience of fuel cell power generation and cryogenic reactant supply systems are reviewed, beginning with the state of technology at the conclusion of the Apollo Program. Technology advancements span a period of 10 years from initial definition phase to the most recent space transportation system (STS) flights. The development program encompassed prototype, verification, and qualification hardware, as well as post-STS-1 design improvements. Focus is on the problems encountered, the scientific and engineering approaches employed to meet the technological challenges, and the results obtained. Major technology barriers are discussed, and the evolving technology development paths are traced from their conceptual beginnings to the fully man-rated systems which are now an integral part of the shuttle vehicle.
The optical design and simulation of the collimated solar simulator
NASA Astrophysics Data System (ADS)
Zhang, Jun; Ma, Tao
2018-01-01
The solar simulator is a lighting device that can simulate the solar radiation. It has been widely used in the testing of solar cells, satellite space environment simulation and ground experiment, test and calibration precision of solar sensor. The solar simulator mainly consisted of short—arc xenon lamp, ellipsoidal reflectors, a group of optical integrator, field stop, aspheric folding mirror and collimating reflector. In this paper, the solar simulator's optical system basic size are given by calculation. Then the system is optically modeled with the Lighttools software, and the simulation analysis on solar simulator using the Monte Carlo ray -tracing technique is conducted. Finally, the simulation results are given quantitatively by diagrammatic form. The rationality of the design is verified on the basis of theory.
Ren, Ling; Xu, Lu; Feng, Jingwen; Zhang, Yang; Yang, Ke
2012-05-01
A novel 316L type Cu-bearing stainless steel was developed in present work, aiming at reducing the occurrence of the in-stent restenosis after implantations of coronary stents, through trace amount of Cu release from surface of the steel in body fluid. It was found that there was a trace amount of Cu released from the Cu-bearing steel in a simulated body fluid, with no cytotoxicity. All the in vitro experimental results proved that this Cu-bearing steel could not only inhibit the proliferation of vascular smooth muscle cells, reducing the formation of thrombosis, which are the main reasons for happening of the in-stent restenosis, but also promote the proliferation of vascular endothelial cells needed for the revascularization, showing that this novel steel is prospective to be a new material for manufacturing coronary stents with function of reducing the in-stent restenosis.
Coming to Critical Pedagogy: A Marxist Autobiography in the History of Higher Education
ERIC Educational Resources Information Center
Malott, Curry Stephenson
2014-01-01
In this essay Malott traces his journey to critical pedagogy focusing on a significant element of his family's ethnic and class background and its connection to his own educational experiences from public schooling to university. Drawing on Marx's historical discussions at the end of Volume 1 of "Capital" Malott traces his own…
NASA Technical Reports Server (NTRS)
Atlas, E.; Ridley, B.; Walega, J.; Greenberg, J.; Kok, G.; Staffelbach, T.; Schauffler, S.; Lind, J.; Huebler, G.; Norton, R.
1996-01-01
During October 19-20, 1991, one flight of the NASA Global Tropospheric Experiment (GTE) Pacific Exploratory Mission (PEM-West A) mission was conducted near Hawaii as an intercomparison with ground-based measurements of the Mauna Loa Observatory Photochemistry Experiment (MLOPEX 2) and the NOAA Climate Modeling and Diagnostics Laboratory (CMDL). Ozone, reactive nitrogen species, peroxides, hydrocarbons, and halogenated hydrocarbons were measured by investigators aboard the DC-8 aircraft and at the ground site. Lidar cross sections of ozone revealed a complex air mass structure near the island of Hawaii which was evidenced by large variation in some trace gas mixing ratios. This variation limited the time and spatial scales for direct measurement intercomparisons. Where differences occurred between measurements in the same air masses, the intercomparison suggested that biases for some trace gases was due to different calibration scales or, in some cases, instrumental or sampling biases. Relatively large uncertainties were associated with those trace gases present in the low parts per trillion by volume range. Trace gas correlations were used to expand the scope of the intercomparison to identify consistent trends between the different data sets.
Effects of Cabin Upsets on Adsorption Columns for Air Revitalization
NASA Technical Reports Server (NTRS)
LeVan, Douglas
1999-01-01
The National Aeronautics and Space Administration (NASA) utilizes adsorption technology as part of contaminant removal systems designed for long term missions. A variety of trace contaminants can be effectively removed from gas streams by adsorption onto activated carbon. An activated carbon adsorption column meets NASA's requirements of a lightweight and efficient means of controlling trace contaminant levels aboard spacecraft and space stations. The activated carbon bed is part of the Trace Contaminant Control System (TCCS) which is utilized to purify the cabin atmosphere. TCCS designs oversize the adsorption columns to account for irregular fluctuations in cabin atmospheric conditions. Variations in the cabin atmosphere include changes in contaminant concentrations, temperature, and relative humidity. Excessively large deviations from typical conditions can result from unusual crew activity, equipment malfunctions, or even fires. The research carried out under this award focussed in detail on the effects of cabin upsets on the performance of activated carbon adsorption columns. Both experiments and modeling were performed with an emphasis on the roll of a change in relative humidity on adsorption of trace contaminants. A flow through fixed-bed apparatus was constructed at the NASA Ames Research Center, and experiments were performed there. Modeling work was performed at the University of Virginia.
The role of endocytic pathways in cellular uptake of plasma non-transferrin iron
Sohn, Yang-Sung; Ghoti, Hussam; Breuer, William; Rachmilewitz, Eliezer; Attar, Samah; Weiss, Guenter; Cabantchik, Z. Ioav
2012-01-01
Background In transfusional siderosis, the iron binding capacity of plasma transferrin is often surpassed, with concomitant generation of non-transferrin-bound iron. Although implicated in tissue siderosis, non-transferrin-bound iron modes of cell ingress remain undefined, largely because of its variable composition and association with macromolecules. Using fluorescent tracing of labile iron in endosomal vesicles and cytosol, we examined the hypothesis that non-transferrin-bound iron fractions detected in iron overloaded patients enter cells via bulk endocytosis. Design and Methods Fluorescence microscopy and flow cytometry served as analytical tools for tracing non-transferrin-bound iron entry into endosomes with the redox-reactive macromolecular probe Oxyburst-Green and into the cytosol with cell-laden calcein green and calcein blue. Non-transferrin-bound iron-containing media were from sera of polytransfused thalassemia major patients and model iron substances detected in thalassemia major sera; cell models were cultured macrophages, and cardiac myoblasts and myocytes. Results Exposure of cells to ferric citrate together with albumin, or to non-transferrin-bound iron-containing sera from thalassemia major patients caused an increase in labile iron content of endosomes and cytosol in macrophages and cardiac cells. This increase was more striking in macrophages, but in both cell types was largely reduced by co-exposure to non-transferrin-bound iron-containing media with non-penetrating iron chelators or apo-transferrin, or by treatment with inhibitors of endocytosis. Endosomal iron accumulation traced with calcein-green was proportional to input non-transferrin-bound iron levels (r2=0.61) and also preventable by pre-chelation. Conclusions Our studies indicate that macromolecule-associated non-transferrin-bound iron can initially gain access into various cells via endocytic pathways, followed by iron translocation to the cytosol. Endocytic uptake of plasma non-transferrin-bound iron is a possible mechanism that can contribute to iron loading of cell types engaged in bulk/adsorptive endocytosis, highlighting the importance of its prevention by iron chelation. PMID:22180428
Liao, Ting T; Wang, Lei; Jia, Ru W; Fu, Xiao H; Chua, Hong
2014-01-01
Membrane damage related to morphological change in Vero cells is a sensitive index of the composite biotoxicity of trace lipophilic chemicals. However, judging whether the morphological change in Vero cells happens and its ratio are difficult because it is not a quantitative characteristic. To find biomarkers of cell morphological change for quantitatively representing the ratio of morphological changed cell, the mechanism of cell membrane damage driven by typical lipophilic chemicals, such as trichlorophenol (TCP) and perfluorooctanesulphonate (PFOS), was explored. The ratio of morphologically changed cells generally increased with increased TCP or PFOS concentrations, and the level of four major components of phospholipids varied with concentrations of TCP or PFOS, but only the ratio of phosphatidylcholine (PC)/phosphatidylethanolamine (PE) decreased regularly as TCP or PFOS concentrations increased. Analysis of membrane proteins showed that the level of vimentin in normal cell membranes is high, while it decreases or vanishes after TCP exposure. These variations in phospholipid and membrane protein components may result in membrane leakage and variation in rigid structure, which leads to changes in cell morphology. Therefore, the ratio of PC/PE and amount of vimentin may be potential biomarkers for representing the ratio of morphological changed Vero cell introduced by trace lipophilic compounds, thus their composite bio-toxicity.
Cairo, Mitchell S; Rocha, Vanderson; Gluckman, Eliane; Hale, Gregory; Wagner, John
2008-01-01
Allogeneic stem cell transplantation has been demonstrated to be curative in a wide variety of pediatric malignant and nonmalignant diseases, and can be traced back over 50 years ago to the original report of Thomas et al. HLA matched sibling donors have been the gold standard for pediatric recipients requiring allogeneic donors for both nonmalignant and malignant conditions. However, only 25% of potential pediatric recipients possesses an HLA-matched sibling donor, and the frequency is even less in those with genetic nonmalignant conditions because of genetically affected other siblings within the family. Therefore, 75% to 90% of potential pediatric recipients require alternative allogeneic donor cells for treatment of their underlying conditions. Potential alternative allogeneic donor sources include unrelated cord blood donors, unrelated adult donors, and haploidentical family donors. In this article we review the experience of both unrelated cord blood donor and haploidentical family donor transplants in selected pediatric malignant and nonmalignant conditions.
NASA Technical Reports Server (NTRS)
Sears, B.; Narayanan, R.; Anderson, T. J.; Fripp, A. L.
1992-01-01
An electrochemical titration method was used to investigate the dynamic states in a cylindrical layer of convecting tin. The liquid tin was contained in a cell, with curved boundaries made of quartz and flat boundaries made of a solid state electrolyte - yttria-stabilized zirconia (YSZ). The electrolyte acted as a window through which a trace amount of oxygen could be pumped in or out by the application of a constant voltage. The concentration at the YSZ interface was monitored by operating the electrochemical cell in the galvanic mode. Experimentally determined effective diffusivities of oxygen were compared with the molecular diffusivity. Dynamic states in the convective flow were thus inferred. Temperature measurements were simultaneously made in order to identify the onset of oscillations from a steady convective regime. The experiments were conducted for two different aspect ratios for various imposed temperature gradients and two different orientations with respect to gravity. Transcritical states were identified and comparison to two-dimensional numerical models were made.
Haldipur, Parthiv; Dang, Derek; Aldinger, Kimberly A; Janson, Olivia K; Guimiot, Fabien; Adle-Biasette, Homa; Dobyns, William B; Siebert, Joseph R; Russo, Rosa; Millen, Kathleen J
2017-01-16
FOXC1 loss contributes to Dandy-Walker malformation (DWM), a common human cerebellar malformation. Previously, we found that complete Foxc1 loss leads to aberrations in proliferation, neuronal differentiation and migration in the embryonic mouse cerebellum (Haldipur et al., 2014). We now demonstrate that hypomorphic Foxc1 mutant mice have granule and Purkinje cell abnormalities causing subsequent disruptions in postnatal cerebellar foliation and lamination. Particularly striking is the presence of a partially formed posterior lobule which echoes the posterior vermis DW 'tail sign' observed in human imaging studies. Lineage tracing experiments in Foxc1 mutant mouse cerebella indicate that aberrant migration of granule cell progenitors destined to form the posterior-most lobule causes this unique phenotype. Analyses of rare human del chr 6p25 fetal cerebella demonstrate extensive phenotypic overlap with our Foxc1 mutant mouse models, validating our DWM models and demonstrating that many key mechanisms controlling cerebellar development are likely conserved between mouse and human.
Generation and characterization of Lhx9 – GFPCreERT2 knock-in mouse line
Xie, Xiaoling; Deng, Min; Gan, Lin
2014-01-01
Summary LHX9 is a LIM-homeodomain transcription factor essential for the development of gonads, spinal cord interneurons, and thalamic neurons to name a few. We recently reported the expression of LHX9 in retinal amacrine cells during development. In this study, we generated an Lhx9 - GFPCreERT2 (GCE) knock-in mouse line by knocking-in a GCE cassette at the Lhx9 locus, thus inactivating endogenous Lhx9. Lhx9GCE/+ mice were viable, fertile, and displayed no overt phenotypical characteristics. Lhx9GCE/GCE mice were all phenotypically female, smaller in size, viable, but infertile. The specificity and efficacy of the Lhx9-GCE mouse line was verified by crossing it to a Rosa26 - tdTomato reporter mouse line, which reveals the Cre recombinase activities in retinal amacrine cells, developing limbs, testis, hippocampal neurons, thalamic neurons, and cerebellar neurons. Taken together, the Lhx9-GCE mouse line could serve as a beneficial tool for lineage tracing and gene manipulation experiments. PMID:25112520
NASA Astrophysics Data System (ADS)
Mojiri, Ahmad; Stanley, Cameron; Rosengarten, Gary
2015-09-01
Hybrid photovoltaic/thermal (PV-T) solar collectors are capable of delivering heat and electricity concurrently. Implementing such receivers in linear concentrators for high temperature applications need special considerations such as thermal decoupling of the photovoltaic (pv) cells from the thermal receiver. Spectral beam splitting of concentrated light provides an option for achieving this purpose. In this paper we introduce a relatively simple hybrid receiver configuration that spectrally splits the light between a high temperature thermal fluid and silicon pv cells using volumetric light filtering by semi-conductor doped glass and propylene glycol. We analysed the optical performance of this device theoretically using ray tracing and experimentally through the construction and testing of a full scale prototype. The receiver was mounted on a commercial parabolic trough concentrator in an outdoor experiment. The prototype receiver delivered heat and electricity at total thermal efficiency of 44% and electrical efficiency of 3.9% measured relative to the total beam energy incident on the primary mirror.
Genome-Wide RNAi Ionomics Screen Reveals New Genes and Regulation of Human Trace Element Metabolism
Malinouski, Mikalai; Hasan, Nesrin M.; Zhang, Yan; Seravalli, Javier; Lin, Jie; Avanesov, Andrei; Lutsenko, Svetlana; Gladyshev, Vadim N.
2017-01-01
Trace elements are essential for human metabolism and dysregulation of their homeostasis is associated with numerous disorders. Here we characterize mechanisms that regulate trace elements in human cells by designing and performing a genome-wide high-throughput siRNA/ionomics screen, and examining top hits in cellular and biochemical assays. The screen reveals high stability of the ionomes, especially the zinc ionome, and yields known regulators and novel candidates. We further uncover fundamental differences in the regulation of different trace elements. Specifically, selenium levels are controlled through the selenocysteine machinery and expression of abundant selenoproteins; copper balance is affected by lipid metabolism and requires machinery involved in protein trafficking and posttranslational modifications; and the iron levels are influenced by iron import and expression of the iron/heme-containing enzymes. Our approach can be applied to a variety of disease models and/or nutritional conditions, and the generated dataset opens new directions for studies of human trace element metabolism. PMID:24522796
Wang, W.-X.; Fisher, N.S.; Luoma, S. N.
1995-01-01
Pulse-chase feeding and multi-labeled radiotracer techniques were employed to measure the assimilation of 6 trace elements (110mAg, 241Am, 109Cd, 57Co, 75Se and 65Zn) from ingested diatoms in the mussel Mytilus edulis feeding at different rates (0.1, 0.49 and 1.5 mg dry wt h-1). Uniformly radiolabeled diatoms Thalassiosira pseudonana were fed to mussels for 0.5 h, and the behavior of the radiotracers in individual mussels was followed for 96 h in a depuration seawater system. Assimilation efficiency (AE) of each element declined with increasing ingestion rate and increased with gut passage time. The importance of extracellular digestion relative to intracellular digestion increased with ingestion activity, which, when coupled with a decline in AE, suggested that extracellular digestion is less efficient in metal absorption. Zn assimilation was most affected by ingestion rate, suggesting that AE may play a role in the physiological regulation of this metal in M. edulis. In an experiment to simulate the effects of an acidic gut, lowered pH (5.5) enhanced the release of elements from intact diatom cells, especially at low particle concentration. These results indicate that both feeding components of the mussel (i.e. gut passage time, digestive partitioning) and metal chemistry (i.e. metal release at lowered pH within the bivalve gut) are responsible for the difference in the assimilation of trace metals at different food quantities observed in mussels.
Galeazzi, Juan M.; Navajas, Joaquín; Mender, Bedeho M. W.; Quian Quiroga, Rodrigo; Minini, Loredana; Stringer, Simon M.
2016-01-01
ABSTRACT Neurons have been found in the primate brain that respond to objects in specific locations in hand-centered coordinates. A key theoretical challenge is to explain how such hand-centered neuronal responses may develop through visual experience. In this paper we show how hand-centered visual receptive fields can develop using an artificial neural network model, VisNet, of the primate visual system when driven by gaze changes recorded from human test subjects as they completed a jigsaw. A camera mounted on the head captured images of the hand and jigsaw, while eye movements were recorded using an eye-tracking device. This combination of data allowed us to reconstruct the retinal images seen as humans undertook the jigsaw task. These retinal images were then fed into the neural network model during self-organization of its synaptic connectivity using a biologically plausible trace learning rule. A trace learning mechanism encourages neurons in the model to learn to respond to input images that tend to occur in close temporal proximity. In the data recorded from human subjects, we found that the participant’s gaze often shifted through a sequence of locations around a fixed spatial configuration of the hand and one of the jigsaw pieces. In this case, trace learning should bind these retinal images together onto the same subset of output neurons. The simulation results consequently confirmed that some cells learned to respond selectively to the hand and a jigsaw piece in a fixed spatial configuration across different retinal views. PMID:27253452
Galeazzi, Juan M; Navajas, Joaquín; Mender, Bedeho M W; Quian Quiroga, Rodrigo; Minini, Loredana; Stringer, Simon M
2016-01-01
Neurons have been found in the primate brain that respond to objects in specific locations in hand-centered coordinates. A key theoretical challenge is to explain how such hand-centered neuronal responses may develop through visual experience. In this paper we show how hand-centered visual receptive fields can develop using an artificial neural network model, VisNet, of the primate visual system when driven by gaze changes recorded from human test subjects as they completed a jigsaw. A camera mounted on the head captured images of the hand and jigsaw, while eye movements were recorded using an eye-tracking device. This combination of data allowed us to reconstruct the retinal images seen as humans undertook the jigsaw task. These retinal images were then fed into the neural network model during self-organization of its synaptic connectivity using a biologically plausible trace learning rule. A trace learning mechanism encourages neurons in the model to learn to respond to input images that tend to occur in close temporal proximity. In the data recorded from human subjects, we found that the participant's gaze often shifted through a sequence of locations around a fixed spatial configuration of the hand and one of the jigsaw pieces. In this case, trace learning should bind these retinal images together onto the same subset of output neurons. The simulation results consequently confirmed that some cells learned to respond selectively to the hand and a jigsaw piece in a fixed spatial configuration across different retinal views.
Wingenter, Oliver W; Haase, Karl B; Strutton, Peter; Friederich, Gernot; Meinardi, Simone; Blake, Donald R; Rowland, F Sherwood
2004-06-08
Oceanic iron (Fe) fertilization experiments have advanced the understanding of how Fe regulates biological productivity and air-sea carbon dioxide (CO(2)) exchange. However, little is known about the production and consumption of halocarbons and other gases as a result of Fe addition. Besides metabolizing inorganic carbon, marine microorganisms produce and consume many other trace gases. Several of these gases, which individually impact global climate, stratospheric ozone concentration, or local photochemistry, have not been previously quantified during an Fe-enrichment experiment. We describe results for selected dissolved trace gases including methane (CH(4)), isoprene (C(5)H(8)), methyl bromide (CH(3)Br), dimethyl sulfide, and oxygen (O(2)), which increased subsequent to Fe fertilization, and the associated decreases in concentrations of carbon monoxide (CO), methyl iodide (CH(3)I), and CO(2) observed during the Southern Ocean Iron Enrichment Experiments.
Footprints of "experiment" in early Arabic optics.
Kheirandish, Elaheh
2009-01-01
This study traces the early developments of the concept of experiment with a view of extending the subject in both content and approach. It extends the content of the subject slightly backward, prior to the methodological breakthroughs of the Optics of Ibn al-Haytham (Alhazen or Alhacen, d. ca. 1040), which are credited as a "significant landmark in the history of experimental science." And it extends the approach to the subject slightly forward, from the premise that early science was "largely carried out in books," to a close examination of the books through which the footprints of'experiment' may be traced. The point of departure is the Optics of Ahmad ibn 'Isă, a revealing text for the early developments of concepts such as 'demonstration' and 'experiment', and one through which some modern discussions are examined and extended with reference to this and other historical sources.
Temporal patterns of inputs to cerebellum necessary and sufficient for trace eyelid conditioning.
Kalmbach, Brian E; Ohyama, Tatsuya; Mauk, Michael D
2010-08-01
Trace eyelid conditioning is a form of associative learning that requires several forebrain structures and cerebellum. Previous work suggests that at least two conditioned stimulus (CS)-driven signals are available to the cerebellum via mossy fiber inputs during trace conditioning: one driven by and terminating with the tone and a second driven by medial prefrontal cortex (mPFC) that persists through the stimulus-free trace interval to overlap in time with the unconditioned stimulus (US). We used electric stimulation of mossy fibers to determine whether this pattern of dual inputs is necessary and sufficient for cerebellar learning to express normal trace eyelid responses. We find that presenting the cerebellum with one input that mimics persistent activity observed in mPFC and the lateral pontine nuclei during trace eyelid conditioning and another that mimics tone-elicited mossy fiber activity is sufficient to produce responses whose properties quantitatively match trace eyelid responses using a tone. Probe trials with each input delivered separately provide evidence that the cerebellum learns to respond to the mPFC-like input (that overlaps with the US) and learns to suppress responding to the tone-like input (that does not). This contributes to precisely timed responses and the well-documented influence of tone offset on the timing of trace responses. Computer simulations suggest that the underlying cerebellar mechanisms involve activation of different subsets of granule cells during the tone and during the stimulus-free trace interval. These results indicate that tone-driven and mPFC-like inputs are necessary and sufficient for the cerebellum to learn well-timed trace conditioned responses.
Mutant p53 establishes targetable tumor dependency by promoting unscheduled replication
Singh, Shilpa; Vaughan, Catherine A.; Frum, Rebecca A.; Grossman, Steven R.; Deb, Sumitra
2017-01-01
Gain-of-function (GOF) p53 mutations are observed frequently in most intractable human cancers and establish dependency for tumor maintenance and progression. While some of the genes induced by GOF p53 have been implicated in more rapid cell proliferation compared with p53-null cancer cells, the mechanism for dependency of tumor growth on mutant p53 is unknown. This report reveals a therapeutically targetable mechanism for GOF p53 dependency. We have shown that GOF p53 increases DNA replication origin firing, stabilizes replication forks, and promotes micronuclei formation, thus facilitating the proliferation of cells with genomic abnormalities. In contrast, absence or depletion of GOF p53 leads to decreased origin firing and a higher frequency of fork collapse in isogenic cells, explaining their poorer proliferation rate. Following genome-wide analyses utilizing ChIP-Seq and RNA-Seq, GOF p53–induced origin firing, micronuclei formation, and fork protection were traced to the ability of GOF p53 to transactivate cyclin A and CHK1. Highlighting the therapeutic potential of CHK1’s role in GOF p53 dependency, experiments in cell culture and mouse xenografts demonstrated that inhibition of CHK1 selectively blocked proliferation of cells and tumors expressing GOF p53. Our data suggest the possibility that checkpoint inhibitors could efficiently and selectively target cancers expressing GOF p53 alleles. PMID:28394262
Design of a Single-Cell Positioning Controller Using Electroosmotic Flow and Image Processing
Ay, Chyung; Young, Chao-Wang; Chen, Jhong-Yin
2013-01-01
The objective of the current research was not only to provide a fast and automatic positioning platform for single cells, but also improved biomolecular manipulation techniques. In this study, an automatic platform for cell positioning using electroosmotic flow and image processing technology was designed. The platform was developed using a PCI image acquisition interface card for capturing images from a microscope and then transferring them to a computer using human-machine interface software. This software was designed by the Laboratory Virtual Instrument Engineering Workbench, a graphical language for finding cell positions and viewing the driving trace, and the fuzzy logic method for controlling the voltage or time of an electric field. After experiments on real human leukemic cells (U-937), the success of the cell positioning rate achieved by controlling the voltage factor reaches 100% within 5 s. A greater precision is obtained when controlling the time factor, whereby the success rate reaches 100% within 28 s. Advantages in both high speed and high precision are attained if these two voltage and time control methods are combined. The control speed with the combined method is about 5.18 times greater than that achieved by the time method, and the control precision with the combined method is more than five times greater than that achieved by the voltage method. PMID:23698272
Design and testing of a uniformly solar energy TIR-R concentration lenses for HCPV systems.
Shen, S C; Chang, S J; Yeh, C Y; Teng, P C
2013-11-04
In this paper, total internal reflection-refraction (TIR-R) concentration (U-TIR-R-C) lens module were designed for uniformity using the energy configuration method to eliminate hot spots on the surface of solar cell and increase conversion efficiency. The design of most current solar concentrators emphasizes the high-power concentration of solar energy, however neglects the conversion inefficiency resulting from hot spots generated by uneven distributions of solar energy concentrated on solar cells. The energy configuration method proposed in this study employs the concept of ray tracing to uniformly distribute solar energy to solar cells through a U-TIR-R-C lens module. The U-TIR-R-C lens module adopted in this study possessed a 76-mm diameter, a 41-mm thickness, concentration ratio of 1134 Suns, 82.6% optical efficiency, and 94.7% uniformity. The experiments demonstrated that the U-TIR-R-C lens module reduced the core temperature of the solar cell from 108 °C to 69 °C and the overall temperature difference from 45 °C to 10 °C, and effectively relative increased the conversion efficiency by approximately 3.8%. Therefore, the U-TIR-R-C lens module designed can effectively concentrate a large area of sunlight onto a small solar cell, and the concentrated solar energy can be evenly distributed in the solar cell to achieve uniform irradiance and effectively eliminate hot spots.
Yu, Xiaoxiao; Chen, Beibei; He, Man; Wang, Han; Hu, Bin
2018-03-01
The quantification of trace Cd and Se in cells incubated with CdSe quantum dots (QDs) is critical to investigate the cytotoxicity of CdSe QDs. In this work, a miniaturized platform, namely chip-based magnetic solid phase microextraction (MSPME) packing with sulfhydryl group functionalized magnetic nanoparticles, was fabricated and combined with inductively coupled plasma mass spectrometry (ICP-MS) for the determination of trace Cd and Se in cells. Under the optimized conditions, the limits of detection (LOD) of the developed chip-based MSPME-ICP-MS system are 2.2 and 21ngL -1 for Cd and Se, respectively. The proposed method is applied successfully to the analysis of total and released small molecular fraction of Cd and Se in Human hepatocellular carcinoma cells (HepG2 cells) incubated with CdSe QDs, and the recoveries for the spiked samples are in the range of 86.0-109%. This method shows great promise to analyze cell samples and the obtained results are instructive to explore the cytotoxicity mechanism of CdSe QDs in cells. Copyright © 2017 Elsevier B.V. All rights reserved.
Academic Odyssey: The Training of an African Sociologist.
ERIC Educational Resources Information Center
Woldemikael, Tekle M.
1988-01-01
Traces the author's experiences in a Eurocentric graduate training program relative to institutional pressures, cultural conflict, interactionism, and U.S. racism. Concludes that experiences caused an intellectual shift to viewpoint of a marginal Black sociologist who focuses upon the experiences and history of persons of African descent in the…
Lin, Yi-Zhen; Ou, Da-Liang; Chang, Hsin-Yuan; Lin, Wei-Yu; Hsu, Chiun
2017-01-01
The family of microRNAs (miRNAs) not only plays an important role in gene regulation but is also useful for the diagnosis of diseases. A reliable method with high sensitivity may allow researchers to detect slight fluctuations in ultra-trace amounts of miRNA. In this study, we propose a sensitive imaging method for the direct probing of miR-10b (miR-10b-3p, also called miR-10b*) and its target (HOXD10 mRNA) in fixed cells based on the specific recognition of molecular beacons combined with highly inclined and laminated optical sheet (HILO) fluorescence microscopy. The designed dye-quencher-labelled molecular beacons offer excellent efficiencies of fluorescence resonance energy transfer that allow us to detect miRNA and the target mRNA simultaneously in hepatocellular carcinoma cells using HILO fluorescence microscopy. Not only can the basal trace amount of miRNA be observed in each individual cell, but the obtained images also indicate that this method is useful for monitoring the fluctuations in ultra-trace amounts of miRNA when the cells are transfected with a miRNA precursor or a miRNA inhibitor (anti-miR). Furthermore, a reasonable causal relation between the miR-10b and HOXD10 expression levels was observed in miR-10b* precursor-transfected cells and miR-10b* inhibitor-transfected cells. The trends of the miRNA alterations obtained using HILO microscopy completely matched the RT-qPCR data and showed remarkable reproducibility (the coefficient of variation [CV] = 0.86%) and sensitivity (<1.0 fM). This proposed imaging method appears to be useful for the simultaneous visualisation of ultra-trace amounts of miRNA and target mRNA and excludes the procedures for RNA extraction and amplification. Therefore, the visualisation of miRNA and the target mRNA should facilitate the exploration of the functions of ultra-trace amounts of miRNA in fixed cells in biological studies and may serve as a powerful tool for diagnoses based on circulating cancer cells. PMID:28989695
From New Haven to Santa Clara: A Historical Perspective on the Milgram Obedience Experiments
ERIC Educational Resources Information Center
Blass, Thomas
2009-01-01
This article traces the history of obedience experiments that have used the Milgram paradigm. It begins with Stanley Milgram's graduate education, showing how some aspects of that experience laid the groundwork for the obedience experiments. It then identifies three factors that led Milgram to study obedience. The underlying principles or messages…
Yu, Qing Cissy; Song, Wenqian; Lai, Dengwen; Zeng, Yi Arial
2017-08-03
Endothelial cells (ECs) are the fundamental building blocks of the vascular architecture and mediate vascular growth and remodeling to ensure proper vessel development and homeostasis. However, studies on endothelial lineage hierarchy remain elusive due to the lack of tools to gain access as well as to directly evaluate their behavior in vivo. To address this shortcoming, a new tissue model to study angiogenesis using the mammary fat pad has been developed. The mammary gland develops mostly in the postnatal stages, including puberty and pregnancy, during which robust epithelium proliferation is accompanied by extensive vascular remodeling. Mammary fat pads provide space, matrix, and rich angiogenic stimuli from the growing mammary epithelium. Furthermore, mammary fat pads are located outside the peritoneal cavity, making them an easily accessible grafting site for assessing the angiogenic potential of exogenous cells. This work also describes an efficient tracing approach using fluorescent reporter mice to specifically label the targeted population of vascular endothelial stem cells (VESCs) in vivo. This lineage tracing method, coupled with subsequent tissue whole-mount microscopy, enable the direct visualization of targeted cells and their descendants, through which the proliferation capability can be quantified and the differentiation commitment can be fate-mapped. Using these methods, a population of bipotent protein C receptor (Procr) expressing VESCs has recently been identified in multiple vascular systems. Procr + VESCs, giving rise to both new ECs and pericytes, actively contribute to angiogenesis during development, homeostasis, and injury repair. Overall, this manuscript describes a new mammary fat pad transplantation and in vivo lineage tracing techniques that can be used to evaluate the stem cell properties of VESCs.
JÄNICKE, MARTINA; RENISCH, BJÖRN; HAMMERSCHMIDT, MATTHIAS
2012-01-01
Grainyhead/CP2 transcription factor family members are widely conserved among the animal kingdom and have been implicated in different developmental processes. Thus far, nothing has been known about their roles in zebrafish. Here we identify seven zebrafish grainyhead-like (grhl) / cp2 genes, with focus on grhl1, which is expressed in the periderm and in epidermal ionocyte progenitors, but downregulated when ionocytes differentiate. In addition, expression was detected in other “non-keratinocyte” cell types of the epidermis, such as pvalb8-expressing cells, which according to our lineage tracing experiments are derived from the same pool of progenitor cells like keratinocytes and ionocytes. Antisense morpholino oligonucleotide-based loss-of-function analysis revealed that grhl1 is dispensable for the development and function of all investigated epidermal cell types, but required as a negative regulator of its own transcription during ionocyte differentiation. Knockdown of the transcription factor Foxi3a, which is expressed in a subset of the grhl1 population, caused a loss of ionocytes and a corresponding increase in the number of pvalb8-expressing cells, while leaving the number of grhl1-positive cells unaltered. We propose that grhl1 is a novel common marker of all or most “non-keratinocyte” epidermal progenitors, and that the sub-functionalisation of these cells is regulated by differential positive and negative effects of Foxi3 factors. PMID:19757382
NASA Astrophysics Data System (ADS)
Woei Leow, Shin; Corrado, Carley; Osborn, Melissa; Isaacson, Michael; Alers, Glenn; Carter, Sue A.
2013-06-01
Luminescent solar concentrators (LSC) collect ambient light from a broad range of angles and concentrate the captured light onto photovoltaic (PV) cells. LSCs with front-facing cells collect direct and indirect sunlight ensuring a gain factor greater than one. The flexible placement and percentage coverage of PV cells on the LSC panel allow for layout adjustments to be made in order to balance re-absorption losses and the level of light concentration desired. A weighted Monte Carlo ray tracing program was developed to study the transport of photons and loss mechanisms in the LSC to aid in design optimization. The program imports measured absorption/emission spectra of an organic luminescent dye (LR305), the transmission coefficient, and refractive index of acrylic as parameters that describe the system. Simulations suggest that for LR305, 8-10 cm of luminescent material surrounding the PV cell yields the highest increase in power gain per unit area of LSC added, thereby determining the ideal spacing between PV cells in the panel. For rectangular PV cells, results indicate that for each centimeter of PV cell width, an additional increase of 0.15 mm to the waveguide thickness is required to efficiently transport photon collected by the LSC to the PV cell with minimal loss.
Silica-coated titania and zirconia colloids for subsurface transport field experiments
Ryan, Joseph N.; Elimelech, Menachem; Baeseman, Jenny L.; Magelky, Robin D.
2000-01-01
Silica-coated titania (TiO2) and zirconia (ZrO2) colloids were synthesized in two sizes to provide easily traced mineral colloids for subsurface transport experiments. Electrophoretic mobility measurements showed that coating with silica imparted surface properties similar to pure silica to the titania and zirconia colloids. Measurements of steady electrophoretic mobility and size (by dynamic light scattering) over a 90-day period showed that the silica-coated colloids were stable to aggregation and loss of coating. A natural gradient field experiment conducted in an iron oxide-coated sand and gravel aquifer also showed that the surface properties of the silica-coated colloids were similar. Colloid transport was traced at μg L-1 concentrations by inductively coupled plasma-atomic emission spectroscopy measurement of Ti and Zr in acidified samples.
1969-09-01
uses a COMMON block of 663 cells . PLOTT works from a table of data, and assigns to each trace (up to 6 traces) a preset character. Once yoor data is...ALFA 000777 1 ~L P M A LP A U VU I 00) EL4 ’ I’LAC 4-43 FLIER slU.-r t;IINk fLIE" (5,LP.KONUA IlEI 335 50C k k LJ~C I~ N’.12 5 ExTEN, REVISION of...FLAG, PLACLD IN CumM"O. 753 C N3.0,0 AS RUN OAIE (IF BLANK* TAKEN FROM SYSTEM C DATE CELL ) ?SS C e1-63 13 EXECUTION TIME LIMIT IN MINUTES (IF ?b6 C
Adriaenssens, Alice; Lam, Brian Yee Hong; Billing, Lawrence; Skeffington, Katie; Sewing, Sabine
2015-01-01
The stomach epithelium contains a myriad of enteroendocrine cells that modulate a range of physiological functions, including postprandial secretion of regulatory peptides, gastric motility, and nutrient absorption. Somatostatin (SST)-producing D-cells are present in the oxyntic and pyloric regions of the stomach, and provide a tonic inhibitory tone that regulates activity of neighboring enteroendocrine cells and gastric acid secretion. Cellular mechanisms underlying the effects of regulatory factors on gastric D-cells are poorly defined due to problems in identifying primary D-cells, and uncertainty remains about which stimuli influence D-cells directly. In this study, we introduce a transgenic mouse line, SST-Cre, which upon crossing with Cre reporter strains, facilitates the identification and purification of gastric D-cells, or cell-specific expression of genetically encoded calcium indicators. Populations of D-cells from the gastric antrum and corpus were isolated and analyzed by RNA sequencing and quantitative RT-PCR. The expression of hormones, hormone receptors, neurotransmitter receptors, and nutrient receptors was quantified. Pyy, Gipr, Chrm4, Calcrl, Taar1, and Casr were identified as genes that are highly enriched in D-cells compared with SST-negative cells. Hormone secretion assays performed in mixed gastric epithelial cultures confirmed that SST secretion is regulated by incretin hormones, cholecystokinin, acetylcholine, vasoactive intestinal polypeptide, calcitonin gene-related polypeptide, oligopetides, and trace amines. Cholecystokinin and oligopeptides elicited increases in intracellular calcium in single-cell imaging experiments performed using cultured D-cells. Our data provide the first transcriptomic analysis and functional characterization of gastric D-cells, and identify regulatory pathways that underlie the direct detection of stimuli by this cell type. PMID:26241122
NASA Astrophysics Data System (ADS)
Wang, W.; Finlayson-Pitts, B. J.
2003-01-01
The ICP AES experiment reported here is suitable for use in a junior- or senior-level undergraduate instrumental analysis laboratory. The objective of this experiment is to analyze trace metals present in cigarette tobacco, the cigarette filter, and the ash obtained when the cigarette is burned. Two different brands of cigarettes, one with and one without a filter, were used. The filter was analyzed before and after smoke was drawn through it. The trace metals were extracted using concentrated nitric acid at room temperature and at 100 °C respectively, to test the extraction efficiency. Some tobacco samples were spiked with ZnCl2 and FeCl3 to assess the efficiency of the recovery. Zinc and iron are shown to be present in tobacco, filter, and ash, while chromium was above the detection limit only in the ash. These metals are concentrated in the ash compared to the tobacco by factors of ˜4 (Zn), 12 17 (Fe), and ≥ 2 (Cr). If sufficient laboratory time is available, this experiment could be paired with one using atomic absorption (AA) to demonstrate the advantages and disadvantages of ICP when compared to AA.
Antoniotti, M; Park, F; Policriti, A; Ugel, N; Mishra, B
2003-01-01
The analysis of large amounts of data, produced as (numerical) traces of in vivo, in vitro and in silico experiments, has become a central activity for many biologists and biochemists. Recent advances in the mathematical modeling and computation of biochemical systems have moreover increased the prominence of in silico experiments; such experiments typically involve the simulation of sets of Differential Algebraic Equations (DAE), e.g., Generalized Mass Action systems (GMA) and S-systems. In this paper we reason about the necessary theoretical and pragmatic foundations for a query and simulation system capable of analyzing large amounts of such trace data. To this end, we propose to combine in a novel way several well-known tools from numerical analysis (approximation theory), temporal logic and verification, and visualization. The result is a preliminary prototype system: simpathica/xssys. When dealing with simulation data simpathica/xssys exploits the special structure of the underlying DAE, and reduces the search space in an efficient way so as to facilitate any queries about the traces. The proposed system is designed to give the user possibility to systematically analyze and simultaneously query different possible timed evolutions of the modeled system.
NASA Astrophysics Data System (ADS)
González-García, Diego; Zezza, Angela; Behrens, Harald; Vetere, Francesco; Petrelli, Maurizio; Morgavi, Daniele; Perugini, Diego
2016-04-01
New melt injection into a shallow magma chamber is regarded as one of the potential triggers for explosive volcanic eruptions. Chemical diffusion occurring between the two mixing melts is a time-dependent process, and thus has the potential to shed light on the timescales involved in magma mixing events leading to an eruption. In order to achieve this, a complete database of diffusion coefficients in natural melts is a necessary prerequisite. We have carried out a set of 12 diffusion couple experiments in order to determine diffusion coefficients (D) of major and trace elements in two natural silicate melts. Two end-members from the Vulcano island (Aeolian archipelago, Italy) have been chosen for the experiments: a shoshonite (Vulcanello lava platform) and a rhyolitic obsidian (Pietre Cotte lava flow, La Fossa cone). Glasses from each end-member with added water contents of 0 wt%, 1 wt% and 2 wt% were produced in an Internally Heated Pressure Vessel (IHPV). Two glass cylinders with similar water content but different base composition are inserted in Au-Pd capsules and experiments are run in the IHPV at 1200° C with pressure from 0.5 to 3 kbar. Experiment capsules are rapidly quenched and analyzed by FTIR, EPMA and LA-ICP-MS for H2O, major and trace elements, respectively, along 2 mm linear profiles extending across the interface. A Boltzmann-Matano approach is used to obtain concentration-dependent diffusivities. The obtained concentration-distance profiles are asymmetric and extend deeper into the shoshonite relative to the rhyolite, indicating that diffusion is slower in the latter. Results show that diffusivities are notably accelerated by the presence of H2O in the melt. Experiments performed by using water-free glass show diffusivities one order of magnitude lower compared to glasses containing up to 2 wt% H2O. The effect of pressure, in the investigated range, is negligible and falls within measurement error. Among major elements, Si and Ti are the slowest diffusing components, while Na is the fastest. Uphill diffusion minima are observed in Al, Na and some trace elements (Y, Nb, Pb). In contrast to other trace elements, light REE show prominent minima next to the interface between the two melts, with the minimum depth diminishing towards HREE.
Rabies Tracing of Birthdated Dentate Granule Cells in Rat Temporal Lobe Epilepsy
Du, Xi; Zhang, Helen; Parent, Jack M.
2017-01-01
Objective To understand how monosynaptic inputs onto adult-born dentate granule cells (DGCs) are altered in experimental mesial temporal lobe epilepsy (mTLE) and whether their integration differs from early-born DGCs that are mature at the time of epileptogenesis. Methods A dual-virus tracing strategy combining retroviral birthdating with rabies virus-mediated putative retrograde trans-synaptic tracing was used to identify and compare presynaptic inputs onto adult- and early-born DGCs in the rat pilocarpine model of mTLE. Results Our results demonstrate that hilar ectopic DGCs preferentially synapse onto adult-born DGCs after pilocarpine-induced status epilepticus (SE) while normotopic DGCs synapse onto both adult- and early-born DGCs. We also find that parvalbumin+ and somatostatin+ interneuron inputs are greatly diminished onto early-born DGCs after SE. However, somatostatin+ interneuron inputs onto adult-born DGCs are maintained, likely due to preferential sprouting. Intriguingly, CA3 pyramidal cell backprojections that specifically target adult-born DGCs arise in the epileptic brain, while axons of interneurons and pyramidal cells in CA1 appear to sprout across the hippocampal fissure to preferentially synapse onto early-born DGCs. Interpretation These data support the presence of substantial hippocampal circuit remodeling after an epileptogenic insult that generates prominent excitatory monosynaptic inputs, both local recurrent and widespread feedback loops, involving DGCs. Both adult- and early-born DGCs are targets of new inputs from other DGCs as well as from CA3 and CA1 pyramidal cells after pilocarpine-treatment, changes that likely contribute to epileptogenesis in experimental mTLE. PMID:28470680
ERIC Educational Resources Information Center
Fields, Lanny; Doran, Erica; Marroquin, Michael
2009-01-01
Three experiments identified factors that did and did not enhance the formation of two-node four-member equivalence classes when training and testing were conducted with trials presented in a trace stimulus pairing two-response (SP2R) format. All trials contained two separately presented stimuli. Half of the trials, called within-class trials,…
Parallel Symmetric Eigenvalue Problem Solvers
2015-05-01
tutoring, and mentoring experience as an undergraduate. Last but not least, I thank my family for their love and support. v TABLE OF CONTENTS Page LIST...34 4.6.2 Choice of the Ritz shifts . . . . . . . . . . . . . . . . . . . . 38 4.7 Relationship between TraceMin and...which are determined by the Ritz values of the matrix pencil. We conclude with a discussion of the relationship between TraceMin and simultaneous
LBA-ECO TG-07 Soil Trace Gas Flux and Root Mortality, Tapajos National Forest
R.K. Varner; M.M. Keller
2009-01-01
This data set reports the results of an experiment that tested the short-term effects of root mortality on the soil-atmosphere fluxes of nitrous oxide, nitric oxide, methane, and carbon dioxide in a tropical evergreen forest. Weekly trace gas fluxes are provided for treatment and control plots on sand and clay tropical forest soils in two comma separated ASCII files....
ERIC Educational Resources Information Center
Miller, A. Kate
2015-01-01
This study reports on a sentence processing experiment in second language (L2) French that looks for evidence of trace reactivation at clause edge and in the canonical object position in indirect object cleft sentences with complex embedding and cyclic movement. Reaction time (RT) asymmetries were examined among low (n = 20) and high (n = 20)…
ERIC Educational Resources Information Center
Giachero, Marcelo; Bustos, Silvia G.; Calfa, Gaston; Molina, Victor A.
2013-01-01
The present study investigates the fear memory resulting from the interaction of a stressful experience and the retrieval of an established fear memory trace. Such a combination enhanced both fear expression and fear retention in adult Wistar rats. Likewise, midazolam intra-basolateral amygdala (BLA) infusion prior to stress attenuated the…
Evolutionary Consequence of a Trade-Off between Growth and Maintenance along with Ribosomal Damages.
Ying, Bei-Wen; Honda, Tomoya; Tsuru, Saburo; Seno, Shigeto; Matsuda, Hideo; Kazuta, Yasuaki; Yomo, Tetsuya
2015-01-01
Microorganisms in nature are constantly subjected to a limited availability of resources and experience repeated starvation and nutrition. Therefore, microbial life may evolve for both growth fitness and sustainability. By contrast, experimental evolution, as a powerful approach to investigate microbial evolutionary strategies, often targets the increased growth fitness in controlled, steady-state conditions. Here, we address evolutionary changes balanced between growth and maintenance while taking nutritional fluctuations into account. We performed a 290-day-long evolution experiment with a histidine-requiring Escherichia coli strain that encountered repeated histidine-rich and histidine-starved conditions. The cells that experienced seven rounds of starvation and re-feed grew more sustainably under prolonged starvation but dramatically lost growth fitness under rich conditions. The improved sustainability arose from the evolved capability to use a trace amount of histidine for cell propagation. The reduced growth rate was attributed to mutations genetically disturbing the translation machinery, that is, the ribosome, ultimately slowing protein translation. This study provides the experimental demonstration of slow growth accompanied by an enhanced affinity to resources as an evolutionary adaptation to oscillated environments and verifies that it is possible to evolve for reduced growth fitness. Growth economics favored for population increase under extreme resource limitations is most likely a common survival strategy adopted by natural microbes.
McGeown, Sarah P; Gray, Eleanor A; Robinson, Jamey L; Dewhurst, Stephen A
2014-06-01
Two experiments investigated the cognitive skills that underlie children's susceptibility to semantic and phonological false memories in the Deese/Roediger-McDermott procedure (Deese, 1959; Roediger & McDermott, 1995). In Experiment 1, performance on the Verbal Similarities subtest of the British Ability Scales (BAS) II (Elliott, Smith, & McCulloch, 1997) predicted correct and false recall of semantic lures. In Experiment 2, performance on the Yopp-Singer Test of Phonemic Segmentation (Yopp, 1988) did not predict correct recall, but inversely predicted the false recall of phonological lures. Auditory short-term memory was a negative predictor of false recall in Experiment 1, but not in Experiment 2. The findings are discussed in terms of the formation of gist and verbatim traces as proposed by fuzzy trace theory (Reyna & Brainerd, 1998) and the increasing automaticity of associations as proposed by associative activation theory (Howe, Wimmer, Gagnon, & Plumpton, 2009). Copyright © 2014 Elsevier B.V. All rights reserved.
Huertas, Marco A; Schwettmann, Sarah E; Shouval, Harel Z
2016-01-01
The ability to maximize reward and avoid punishment is essential for animal survival. Reinforcement learning (RL) refers to the algorithms used by biological or artificial systems to learn how to maximize reward or avoid negative outcomes based on past experiences. While RL is also important in machine learning, the types of mechanistic constraints encountered by biological machinery might be different than those for artificial systems. Two major problems encountered by RL are how to relate a stimulus with a reinforcing signal that is delayed in time (temporal credit assignment), and how to stop learning once the target behaviors are attained (stopping rule). To address the first problem synaptic eligibility traces were introduced, bridging the temporal gap between a stimulus and its reward. Although, these were mere theoretical constructs, recent experiments have provided evidence of their existence. These experiments also reveal that the presence of specific neuromodulators converts the traces into changes in synaptic efficacy. A mechanistic implementation of the stopping rule usually assumes the inhibition of the reward nucleus; however, recent experimental results have shown that learning terminates at the appropriate network state even in setups where the reward nucleus cannot be inhibited. In an effort to describe a learning rule that solves the temporal credit assignment problem and implements a biologically plausible stopping rule, we proposed a model based on two separate synaptic eligibility traces, one for long-term potentiation (LTP) and one for long-term depression (LTD), each obeying different dynamics and having different effective magnitudes. The model has been shown to successfully generate stable learning in recurrent networks. Although, the model assumes the presence of a single neuromodulator, evidence indicates that there are different neuromodulators for expressing the different traces. What could be the role of different neuromodulators for expressing the LTP and LTD traces? Here we expand on our previous model to include several neuromodulators, and illustrate through various examples how different these contribute to learning reward-timing within a wide set of training paradigms and propose further roles that multiple neuromodulators can play in encoding additional information of the rewarding signal.
Visuomotor learning in cerebellar patients.
Timmann, D; Shimansky, Y; Larson, P S; Wunderlich, D A; Stelmach, G E; Bloedel, J R
1996-11-01
The aim of the present study was to demonstrate that patients with pathology affecting substantial regions of the cerebellum can improve their performance in a series of two-dimensional tracing tasks, thus supporting the view that this type of motor behavior can be acquired even when the integrity of this structure is compromised. Eight patients with chronic, isolated cerebellar lesions and eight age- and sex-matched healthy controls were tested. Three patients had mild, five had moderate upper limb ataxia. The experiment was divided into two parts. In the first, subjects traced an irregularly shaped outline over 20 consecutive trials ('Trace 1' task). Next, subjects were asked to redraw the object without any underlying template as a guide ('Memory 1' task). In the second part of the study, subjects were asked to trace a different, irregularly shaped outline over 20 consecutive trials ('Trace 2' task). Next, they were required to redraw it by memory with its axis rotated 90 degrees ('Memory 2' task). In each of the memory tasks the template was placed over the drawn image after each trial and shown to the subjects. The error of performance was determined by calculating three different measurements, each focused on different aspects of the task. Based on these measurements, the cerebellar patients showed improvement in both memory tasks. In the 'Memory 1' task the calculated error decreased significantly for the patients with mild ataxia. In the 'Memory 2' task all cerebellar patients improved their performance substantially enough to reduce significantly the magnitude of all three error measurements. The experiments demonstrate that patients with cerebellar lesions are capable of improving substantially their performance of a complex motor task involving the recall of memorized shapes and the visuomotor control of a tracing movement.
Interaction of nickel-based SOFC anodes with trace contaminants from coal-derived synthesis gas
NASA Astrophysics Data System (ADS)
Hackett, Gregory Allen
New and efficient methods of producing electrical energy from natural resources have become an important topic for researchers. Integrated gasification and fuel cell (IGFC) systems offer a fuel-flexible, high-efficiency method of energy generation. Specifically, in coal gasification processes, coal can be changed into a high-quality gaseous fuel suitable for feeding solid oxide fuel cells (SOFCs). However, trace species found in coal synthesis gas (syngas) may have a deleterious effect on the performance of nickel-based SOFC anodes. Generally, the cost of removing these species down to parts per million (ppm) levels is high. The purpose of this research is to determine the highest amount of contaminant that results in a low rate (˜1% per 1000 h) of cell performance degradation, allowing the SOFC to produce usable power for 40,000 hours. The cell performance degradation rate was determined for benzene, naphthalene, and mercury-doped syngas based on species concentration. Experimental data are fitted with degradation models to predict cell lifetime behavior. From these results, the minimum coal syngas cleanup required for these trace materials is determined. It is found that for a final cell voltage of 0.6 V, naphthalene and benzene must be cleaned to 360 ppm and less than 150 ppm, respectively. No additional cleaning is required for mercury beyond established environmental standards. Additionally, a detailed attack and recovery mechanism is proposed for the hydrocarbon species and their interaction with the fuel cell. This mechanism is proposed by considering the type of degradation models predicted and how carbon would interact with the Ni-YSZ anode to justify those models. The mechanism postulates that carbon is diffusing into the nickel structure, creating a metal solution. Once the nickel is saturated, the carbon begins to deposit on the nickel surface, reducing the electrode active area. The formation of metal solutions and the deposition of carbon results in reduced cell productivity.
Huang, Miaojun; Li, Tianjie; Pan, Ting; Zhao, Naru; Yao, Yongchang; Zhai, Zhichen; Zhou, Jiaan; Du, Chang; Wang, Yingjun
2016-10-01
Yeast cells have controllable biosorption on metallic ions during metabolism. However, few studies were dedicated to using yeast-regulated biomimetic mineralization process to control the strontium-doped positions in calcium phosphate microcapsules. In this study, the yeast cells were allowed to pre-adsorb strontium ions metabolically and then served as sacrificing template for the precipitation and calcination of mineral shell. The pre-adsorption enabled the microorganism to enrich of strontium ions into the inner part of the microcapsules, which ensured a slow-release profile of the trace element from the microcapsule. The co-culture with human marrow stromal cells showed that gene expressions of alkaline phosphatase and Collagen-I were promoted. The promotion of osteogenic differentiation was further confirmed in the 3D culture of cell-material complexes. The strategy using living microorganism as 'smart doping apparatus' to control incorporation of trace element into calcium phosphate paved a pathway to new functional materials for hard tissue regeneration.
Cell type-specific long-range connections of basal forebrain circuit.
Do, Johnny Phong; Xu, Min; Lee, Seung-Hee; Chang, Wei-Cheng; Zhang, Siyu; Chung, Shinjae; Yung, Tyler J; Fan, Jiang Lan; Miyamichi, Kazunari; Luo, Liqun; Dan, Yang
2016-09-19
The basal forebrain (BF) plays key roles in multiple brain functions, including sleep-wake regulation, attention, and learning/memory, but the long-range connections mediating these functions remain poorly characterized. Here we performed whole-brain mapping of both inputs and outputs of four BF cell types - cholinergic, glutamatergic, and parvalbumin-positive (PV+) and somatostatin-positive (SOM+) GABAergic neurons - in the mouse brain. Using rabies virus -mediated monosynaptic retrograde tracing to label the inputs and adeno-associated virus to trace axonal projections, we identified numerous brain areas connected to the BF. The inputs to different cell types were qualitatively similar, but the output projections showed marked differences. The connections to glutamatergic and SOM+ neurons were strongly reciprocal, while those to cholinergic and PV+ neurons were more unidirectional. These results reveal the long-range wiring diagram of the BF circuit with highly convergent inputs and divergent outputs and point to both functional commonality and specialization of different BF cell types.
Exploring dynamics of molybdate in living animal cells by a genetically encoded FRET nanosensor.
Nakanishi, Yoichi; Iida, Syuntaro; Ueoka-Nakanishi, Hanayo; Niimi, Tomoaki; Tomioka, Rie; Maeshima, Masayoshi
2013-01-01
Molybdenum (Mo) is an essential trace element for almost all living organisms including animals. Mo is used as a catalytic center of molybdo-enzymes for oxidation/reduction reactions of carbon, nitrogen, and sulfur metabolism. Whilst living cells are known to import inorganic molybdate oxyanion from the surrounding environment, the in vivo dynamics of cytosolic molybdate remain poorly understood as no appropriate indicator is available for this trace anion. We here describe a genetically encoded Förester-resonance-energy-transfer (FRET)-based nanosensor composed of CFP, YFP and the bacterial molybdate-sensor protein ModE. The nanosensor MolyProbe containing an optimized peptide-linker responded to nanomolar-range molybdate selectively, and increased YFP:CFP fluorescence intensity ratio by up to 109%. By introduction of the nanosensor, we have been able to successfully demonstrate the real-time dynamics of molybdate in living animal cells. Furthermore, time course analyses of the dynamics suggest that novel oxalate-sensitive- and sulfate-resistant- transporter(s) uptake molybdate in a model culture cell.
Sun, Yanjun; Nguyen, Amanda; Nguyen, Joseph; Le, Luc; Saur, Dieter; Choi, Jiwon; Callaway, Edward M.; Xu, Xiangmin
2014-01-01
Summary We applied a new Cre-dependent, genetically modified rabies-based tracing system to map direct synaptic connections to CA1 excitatory and inhibitory neuron types in mouse hippocampus. We found common inputs to excitatory and inhibitory CA1 neurons from CA3, CA2, entorhinal cortex and the medial septum (MS), and unexpectedly also from the subiculum. Excitatory CA1 neurons receive inputs from both cholinergic and GABAergic MS neurons while inhibitory CA1 neurons receive a great majority of input from GABAergic MS neurons; both cell types also receive weaker input from glutamatergic MS neurons. Comparisons of inputs to CA1 PV+ interneurons versus SOM+ interneurons showed similar strengths of input from the subiculum, but PV+ interneurons receive much stronger input than SOM+ neurons from CA3, entorhinal cortex and MS. Differential input from CA3 to specific CA1 cell types was also demonstrated functionally using laser scanning photostimulation and whole cell recordings. PMID:24656815
NASA Astrophysics Data System (ADS)
Wiche, Oliver; Székely, Balazs; Moschner, Christin; Heilmeier, Hermann
2015-04-01
In recent studies root-soil interactions of white lupine (Lupinus albus L.) have drawn special attention to researchers due to its particularly high potential to increase bioavailability of phosphorous (P) and trace nutrients in soils. In mixed cultures, white lupine has the ability to mobilize P and trace nutrients in soil in excess of its own need and make this excess available for other intercropped companion species. While improved acquisition of P and improved yield parameters have mostly been documented in cereal-lupine intercrops, compared to sole crops, only a few recent studies have evidenced similar effects for trace elements e.g. Fe, Zn and Mn. In this preliminary study we tried to obtain more information about the mobilization of trace elements due to intercropping under field conditions. We hypothesize, that processes that lead to a better acquisition of trace nutrients might also affect other trace elements what could be useful for phytoremediation and phytomining research. Here we report the results of a semi-field experiment were we investigated the effects of an intercropping of white lupine with oat (Avena sativa L.) on the concentrations of trace metals in shoots of oat. We investigated the effects on 12 trace elements, including 4 elements with relevance for plant nutrition (P, Fe, Mn, Zn) and 8 trace elements, belonging to the group of metalloids, lanthanides and actinides with high relevance in phytoremediation (Cd, Pb Th, U) and phytomining research (Sc, La, Nd, Ge). The experiment was carried out on a semi-field lysimer at the off-site soil recycling and remediation center in Hirschfeld (Saxony, Germany). To test the intercropping-dependent mobilization of trace metals in soil and enhanced uptake of elements by oat, white lupine and oat were cultivated on 20 plots (4 m² each) in monocultures and mixed cultures and two different white lupin /oat-ratios (11% and 33%, respectively) applying various treatments. The geometrical arrangement of plots was randomized and every treatment was fivefold replicated. Soil solution was collected weekly with plastic suction cups. Concentrations of trace metals in shoots of oat and soil solution were measured with ICP-MS. As a result, we found that both, concentrations of trace elements in oat plants, as well as the mobility of P and trace metals in soil solution was increased by an intercropping with white lupine. Mixed culture of oat with 11% white lupin significantly increased the concentrations of the trace nutrients Fe, Mn and Zn, as well as the concentrations of the trace metals Pb, La, Nd, Sc, Th and U in tissues of oat. Surprisingly, mixed cultures with 33 % white lupin did not significantly affect trace metal concentrations in oat, what might be the consequence of an increasing competition of roots of white lupin and oat for nutrients and trace metals. In conclusion we found that mixed cultures of white lupin with cereals might be a powerful tool for enhanced phytoremediation and phytomining. However, processes involved in the physiochemical mechanism of element uptake as affected by the oat/white lupin co-cultivation remain unknown and further studies on this topic are planned. These studies have been carried out in the framework of the PhytoGerm project, financed by the Federal Ministry of Education and Research, Germany. The authors are grateful to students and laboratory assistants contributing in the field work and sample preparation.
Hoshino, Osamu
2015-06-01
Perception of supraliminal stimuli might in general be reflected in bursts of action potentials (spikes), and their memory traces could be formed through spike-timing-dependent plasticity (STDP). Memory traces for subliminal stimuli might be formed in a different manner, because subliminal stimulation evokes a fraction (but not a burst) of spikes. Simulations of a cortical neural network model showed that a subliminal stimulus that was too brief (10 msec) to perceive transiently (more than about 500 msec) depolarized stimulus-relevant principal cells and hyperpolarized stimulus-irrelevant principal cells in a subthreshold manner. This led to a small increase or decrease in ongoing-spontaneous spiking activity frequency (less than 1 Hz). Synaptic modification based on STDP during this period effectively enhanced relevant synaptic weights, by which subliminal learning was improved. GABA transporters on GABAergic interneurons modulated local levels of ambient GABA. Ambient GABA molecules acted on extrasynaptic receptors, provided principal cells with tonic inhibitory currents, and contributed to achieving the subthreshold neuronal state. We suggest that ongoing-spontaneous synaptic alteration through STDP following subliminal stimulation may be a possible neuronal mechanism for leaving its memory trace in cortical circuitry. Regulation of local ambient GABA levels by transporter-mediated GABA import and export may be crucial for subliminal learning.
Mobilization of Trace Metals in an Experimental Carbon Sequestration Scenario
NASA Astrophysics Data System (ADS)
Marcon, V.; Kaszuba, J. P.
2012-12-01
Mobilizing trace metals with injection of supercritical CO2 into deep saline aquifers is a concern for geologic carbon sequestration. The potential for leakage from these systems requires an understanding of how injection reservoirs interact with the overlying potable aquifers. Hydrothermal experiments were performed to evaluate metal mobilization and mechanisms of release in a carbonate storage reservoir and at the caprock-reservoir boundary. Experiments react synthetic Desert Creek limestone and/or Gothic Shale, formations in the Paradox Basin, Utah, with brine that is close to equilibrium with these rocks. A reaction temperature of 1600C accelerates the reaction kinetics without changing in-situ water-rock reactions. The experiments were allowed to reach steady state before injecting CO2. Changes in major and trace element water chemistry, dissolved carbon and sulfide, and pH were tracked throughout the experiments. CO2 injection decreases the pH by 1 to 2 units; concomitant mineral dissolution produces elevated Ba, Cu, Fe, Pb, and Zn concentrations in the brine. Concentrations subsequently decrease to approximately steady state values after 120-330 hours, likely due to mineral precipitation as seen in SEM images and predicted by geochemical modeling. In experiments that emulate the caprock-reservoir boundary, final Fe (0.7ppb), an element of secondary concern for the EPA, and Pb (0.05ppb) concentrations exceed EPA limits, whereas Ba (0.140ppb), Cu (48ppb), and Zn (433ppb) values remain below EPA limits. In experiments that simulate deeper reservoir conditions, away from the caprock boundary, final Fe (3.5ppb) and Pb (0.017ppb) values indicate less mobilization than seen at the caprock-reservoir boundary, but values still exceed EPA limits. Barium concentrations always remain below the EPA limit of 2ppb, but are more readily mobilized in experiments replicating deeper reservoir conditions. In both systems, transition elements Cd, Cr, Cu, Pb and Zn behave in a similar manner, increasing in concentration with injection but continually decreasing after about 830 hours until termination of the experiment. SEM images and geochemical models indicate initial dissolution of all rocks and minerals, re-precipitation of Ca-Mg-Fe carbonates and Fe-sulfides, and precipitation of anhydrite in both systems. Calcite dissolves more readily than dolomite in these experiments, but re-precipitates in veins on dolomite. If brines leak from a storage reservoir and mix with a potable aquifer, the experimental results suggest that Ba, Cu, and Zn will not be contaminants of concern. Pb, Fe and As (still under consideration) initially exceed the EPA threshold and may require careful attention in a sequestration scenario. However, experimentally observed trends of decreasing trace metal concentration suggest that these metals could become less of a concern during the life of a carbon repository. Finally, the caprock plays an active role in trace metal mobilization in the system. The caprock provides a source of metals, although subsequent precipitation may remove metals from solution.
Trace element diffusion and kinetic fractionation in wet rhyolitic melt
NASA Astrophysics Data System (ADS)
Holycross, Megan E.; Watson, E. Bruce
2018-07-01
Piston-cylinder experiments were run to determine the chemical diffusivities of 21 trace elements (Sc, V, Y, Zr, Nb, La, Ce, Pr, Nd, Sm, Eu, Gd, Tb, Dy, Ho, Er, Yb, Lu, Hf, Th and U) in hydrous rhyolitic melts at 1 GPa pressure and temperatures from 850 to 1250 °C. Diffusion couple glasses were doped with trace elements in low concentrations to characterize the diffusivities of all cations in a single experiment. Laser ablation ICP-MS was used to evaluate the trace element concentration gradients that developed in the silicate glasses. All calculated diffusion coefficients correspond to the temperature dependence D = D0exp(-Ea/RT). Rhyolite liquids contained either ∼4.1 wt% or ∼6.2 wt% dissolved H2O; separate Arrhenius relationships are produced for each melt composition. Trace element diffusivities in the melt with 6.2 wt% H2O are roughly two times higher than those in the less hydrous melt. Calculated trace element diffusion coefficients cover nearly two orders of magnitude at a given temperature. The high field strength elements are the slowest diffusers, followed by the transition metals and heavy rare earth elements. The light rare earth elements have the fastest diffusion rates in hydrous rhyolitic melt. The measured diffusion coefficients range down to values sufficiently low to preclude diffusive homogenization over geochemically realistic time scales in some cases. The substantial differences in the diffusivities of individual cations may result in fractionated trace element signatures in rhyolite melt pockets. A simple model is used to explore the potential for kinetic fractionation of REE during growth of an apatite crystal in a diffusive boundary layer locally saturated in P2O5. The faster-diffusing light REE are more efficiently transported away from the crystal interface than the slower-moving heavy REE. Diffusion effects will enrich the melt boundary layer in slow-moving HREE relative to the faster LREE. The kinetic fractionation of REE in the melt growth medium will result in a precipitated apatite crystal with a disequilibrium trace element composition.
Laser-induced breakdown spectroscopy for specimen analysis
Kumar, Akshaya; Yu-Yueh, Fang; Burgess, Shane C.; Singh, Jagdish P.
2006-08-15
The present invention is directed to an apparatus, a system and a method for detecting the presence or absence of trace elements in a biological sample using Laser-Induced Breakdown Spectroscopy. The trace elements are used to develop a signature profile which is analyzed directly or compared with the known profile of a standard. In one aspect of the invention, the apparatus, system and method are used to detect malignant cancer cells in vivo.
A miniature microcontroller curve tracing circuit for space flight testing transistors.
Prokop, N; Greer, L; Krasowski, M; Flatico, J; Spina, D
2015-02-01
This paper describes a novel miniature microcontroller based curve tracing circuit, which was designed to monitor the environmental effects on Silicon Carbide Junction Field Effect Transistor (SiC JFET) device performance, while exposed to the low earth orbit environment onboard the International Space Station (ISS) as a resident experiment on the 7th Materials on the International Space Station Experiment (MISSE7). Specifically, the microcontroller circuit was designed to operate autonomously and was flown on the external structure of the ISS for over a year. This curve tracing circuit is capable of measuring current vs. voltage (I-V) characteristics of transistors and diodes. The circuit is current limited for low current devices and is specifically designed to test high temperature, high drain-to-source resistance SiC JFETs. The results of each I-V data set are transmitted serially to an external telemetered communication interface. This paper discusses the circuit architecture, its design, and presents example results.
Metabolic measures of male southern toads (Bufo terrestris) exposed to coal combustion waste
DOE Office of Scientific and Technical Information (OSTI.GOV)
Ward, C.K.; Appel, A.G.; Mendonca, M.T.
2006-03-15
Southern toads (Bufo terrestris) are found in coal fly ash collection basins associated with coal-burning electrical power plants. These basins contain large amounts of trace metals and organisms found in these basins are known to accumulate large quantities of metals. Studies on a variety of organisms exposed to trace metals found that they experience a significant increase in standard metabolic rate. We experimentally exposed southern toads to metal-contaminated sediment and food and measured changes in standard and exercise metabolic rates as well as changes in body, liver and muscle mass, blood glucose, and corticosterone. We found that toads exposed tomore » trace metal contamination gained significantly less mass (18.3%) than control toads (31.3%) when food was limited and experienced significantly decreased RQ after exercise. However, contaminated toads did not experience changes in standard or exercise metabolic rates, plasma glucose levels, and hepatic or muscle percentage indices whether food was limited or not.« less
Takahashi, Y; Endo, Y; Kusaka-Kikushima, A; Nakamaura, S; Nakazawa, Y; Ogi, T; Uryu, M; Tsuji, G; Furue, M; Moriwaki, S
2017-07-01
A certain relationship between XPA gene mutations and the severity of symptoms has been observed in patients with xeroderma pigmentosum group A (XP-A). Patients with mutations within the DNA-binding domain usually exhibit severe symptoms, whereas splicing mutations in the same domain sometimes cause very mild symptoms. This inconsistency can be explained by a small amount of functional XPA protein produced from normally spliced transcripts. We herein report the case of an adult Japanese patient with XP-A with unusually mild symptoms. We identified a homozygous c.529G>A mutation in exon 4 of the XPA gene, which resulted in aberrant splicing with a 29-bp deletion in exon 4 causing a frameshift. Intact mRNA was observable, but a Western blot analysis failed to detect any normal XPA protein. We therefore evaluated the DNA repair capacity in normal cells in which the XPA expression was artificially diminished. The repair capacity was still present in cells with trace levels of the XPA protein. The repair capacity of the cells derived from our patient with mild symptoms was poor by comparison, but still significant compared with that of the cells derived from a patient with XP-A with severe symptoms. These results provide strong evidence that a trace level of XPA protein can still exert a relatively strong repair capacity, resulting in only a mild phenotype. © 2016 British Association of Dermatologists.
Pupil old/new effects reflect stimulus encoding and decoding in short-term memory.
Brocher, Andreas; Graf, Tim
2016-12-01
We conducted five pupil old/new experiments to examine whether pupil old/new effects can be linked to familiarity and/or recollection processes of recognition memory. In Experiments 1-3, we elicited robust pupil old/new effects for legal words and pseudowords (Experiment 1), positive and negative words (Experiment 2), and low-frequency and high-frequency words (Experiment 3). Importantly, unlike for old/new effects in ERPs, we failed to find any effects of long-term memory representations on pupil old/new effects. In Experiment 4, using the words and pseudowords from Experiment 1, participants made lexical decisions instead of old/new decisions. Pupil old/new effects were restricted to legal words. Additionally requiring participants to make speeded responses (Experiment 5) led to a complete absence of old/new effects. Taken together, these data suggest that pupil old/new effects do not map onto familiarity and recollection processes of recognition memory. They rather seem to reflect strength of memory traces in short-term memory, with little influence of long-term memory representations. Crucially, weakening the memory trace through manipulations in the experimental task significantly reduces pupil/old new effects. © 2016 Society for Psychophysiological Research.
NASA Astrophysics Data System (ADS)
Huamán Bustamante, Samuel G.; Cavalcanti Pacheco, Marco A.; Lazo Lazo, Juan G.
2018-07-01
The method we propose in this paper seeks to estimate interface displacements among strata related with reflection seismic events, in comparison to the interfaces at other reference points. To do so, we search for reflection events in the reference point of a second seismic trace taken from the same 3D survey and close to a well. However, the nature of the seismic data introduces uncertainty in the results. Therefore, we perform an uncertainty analysis using the standard deviation results from several experiments with cross-correlation of signals. To estimate the displacements of events in depth between two seismic traces, we create a synthetic seismic trace with an empirical wavelet and the sonic log of the well, close to the second seismic trace. Then, we relate the events of the seismic traces to the depth of the sonic log. Finally, we test the method with data from the Namorado Field in Brazil. The results show that the accuracy of the event estimated depth depends on the results of parallel cross-correlation, primarily those from the procedures used in the integration of seismic data with data from the well. The proposed approach can correctly identify several similar events in two seismic traces without requiring all seismic traces between two distant points of interest to correlate strata in the subsurface.
Xiu, Junshan; Dong, Lili; Qin, Hua; Liu, Yunyan; Yu, Jin
2016-12-01
The detection limit of trace metals in liquids has been improved greatly by laser-induced breakdown spectroscopy (LIBS) using solid substrate. A paper substrate and a metallic substrate were used as a solid substrate for the detection of trace metals in aqueous solutions and viscous liquids (lubricating oils) respectively. The matrix effect on quantitative analysis of trace metals in two types of liquids was investigated. For trace metals in aqueous solutions using paper substrate, the calibration curves established for pure solutions and mixed solutions samples presented large variation on both the slope and the intercept for the Cu, Cd, and Cr. The matrix effects among the different elements in mixed solutions were observed. However, good agreement was obtained between the measured and known values in real wastewater. For trace metals in lubricating oils, the matrix effect between the different oils is relatively small and reasonably negligible under the conditions of our experiment. A universal calibration curve can be established for trace metals in different types of oils. The two approaches are verified that it is possible to develop a feasible and sensitive method with accuracy results for rapid detection of trace metals in industrial wastewater and viscous liquids by laser-induced breakdown spectroscopy. © The Author(s) 2016.
Experiences with hypercube operating system instrumentation
NASA Technical Reports Server (NTRS)
Reed, Daniel A.; Rudolph, David C.
1989-01-01
The difficulties in conceptualizing the interactions among a large number of processors make it difficult both to identify the sources of inefficiencies and to determine how a parallel program could be made more efficient. This paper describes an instrumentation system that can trace the execution of distributed memory parallel programs by recording the occurrence of parallel program events. The resulting event traces can be used to compile summary statistics that provide a global view of program performance. In addition, visualization tools permit the graphic display of event traces. Visual presentation of performance data is particularly useful, indeed, necessary for large-scale parallel computers; the enormous volume of performance data mandates visual display.
NASA Astrophysics Data System (ADS)
Horn, Sabrina; Herrmann, Hartmut
2013-04-01
Mineral dust constitutes one of the largest mass fractions of natural aerosol. Its emission is estimated between 800 - 2000 Tg/a]. The dust is emitted through sand and dust storms in the arid regions of our planet, in particular by the great desserts such as the Sahara. The complex chemical composition of mineral dust is similar to crust material, because the dust is produced by soil erosion. The main components of mineral dust are SiO2 and Al2O3, whereas the active oxides (Fe2O3, TiO2) show some variety in content due to the dust source region. Mineral dust particles can be transported over several 1000 km and during its transport the surface can be changed by the uptake of water vapor and trace gases. On such modified surfaces homo- and heterogeneous reactions can occur. Trace gas uptakes play an important role in atmospheric chemistry as sinks or sources for several gaseous species. Hence, it is necessary to study these reactions. Among several experimental setups, the Knudsen cell is one of the promising tools to study reactive uptakes from the gas phase and the release of products formed by dust surface-mediated reactions. The Knudsen cell, implemented by Golden et al. in 1975, is a high vacuum flow reactor operating under molecular flow conditions, i.e., gas-wall collisions are highly preferred over gas-gas collisions. Despite several Knudsen cell studies examining the reaction between different traces gases and model dust surfaces constituted of not more than a few components, no measurements utilizing collected ambient mineral dust are done so far. For a better understanding of the chemistry on mineral dust surfaces gas uptake measurements will be done with a Knudsen cell. The first measurements are done with isopropanol on TiO2. Afterwards there are studies with different substrates like, Al2O3 (α- and γ-phase), FeO2, Arizona test dust, air collected mineral dust from the Cap Verde islands. In the beginning SO2, NO2 and HNO3 will be used.
Lavery, Danielle L; Martinez, Pierre; Gay, Laura J; Cereser, Biancastella; Novelli, Marco R; Rodriguez-Justo, Manuel; Meijer, Sybren L; Graham, Trevor A; McDonald, Stuart A C; Wright, Nicholas A; Jansen, Marnix
2016-06-01
Barrett's oesophagus commonly presents as a patchwork of columnar metaplasia with and without goblet cells in the distal oesophagus. The presence of metaplastic columnar epithelium with goblet cells on oesophageal biopsy is a marker of cancer progression risk, but it is unclear whether clonal expansion and progression in Barrett's oesophagus is exclusive to columnar epithelium with goblet cells. We developed a novel method to trace the clonal ancestry of an oesophageal adenocarcinoma across an entire Barrett's segment. Clonal expansions in Barrett's mucosa were identified using cytochrome c oxidase enzyme histochemistry. Somatic mutations were identified through mitochondrial DNA sequencing and single gland whole exome sequencing. By tracing the clonal origin of an oesophageal adenocarcinoma across an entire Barrett's segment through a combination of histopathological spatial mapping and clonal ordering, we find that this cancer developed from a premalignant clonal expansion in non-dysplastic ('cardia-type') columnar metaplasia without goblet cells. Our data demonstrate the premalignant potential of metaplastic columnar epithelium without goblet cells in the context of Barrett's oesophagus. Published by the BMJ Publishing Group Limited. For permission to use (where not already granted under a licence) please go to http://www.bmj.com/company/products-services/rights-and-licensing/
Bershetyn, Anna; Odeny, Thomas A; Lyamuya, Rita; Nakiwogga-Muwanga, Alice; Diero, Lameck; Bwana, Mwebesa; Braitstein, Paula; Somi, Geoffrey; Kambugu, Andrew; Bukusi, Elizabeth; Hartogensis, Wendy; Glidden, David V; Wools-Kaloustian, Kara; Yiannoutsos, Constantin; Martin, Jeffrey; Geng, Elvin H
2017-06-01
The effect of tracing human immunodeficiency virus (HIV)-infected patients who are lost to follow-up (LTFU) on reengagement has not been rigorously assessed. We carried out an ex post analysis of a surveillance study in which LTFU patients were randomly selected for tracing to identify the effect of tracing on reengagement. We evaluated HIV-infected adults on antiretroviral therapy who were LTFU (>90 days late for last visit) at 14 clinics in Uganda, Kenya, and Tanzania. A random sample of LTFU patients was selected for tracing by peer health workers. We assessed the effect of selection for tracing using Kaplan-Meier estimates of reengagement among all patients as well as the subset of LTFU patients who were alive, contacted in person by the tracer, and out of care. Of 5781 eligible patients, 991 (17%) were randomly selected for tracing. One year after selection for tracing, 13.3% (95% confidence interval [CI], 11.1%-15.3%) of those selected for tracing returned compared with 10.0% (95% CI, 9.1%-10.8%) of those not randomly selected, an adjusted risk difference of 3.0% (95% CI, .7%-5.3%). Among patients found to be alive, personally contacted, and out of care, tracing increased the absolute probability of return at 1 year by 22% (95% CI, 7.1%-36.2%). The effect of tracing on rate of return to clinic decayed with a half-life of 7.0 days after tracing (95% CI, 2.6 %-12.9%). Tracing interventions increase reengagement, but developing methods for targeting LTFU patients most likely to benefit can make this practice more efficient. © The Author 2017. Published by Oxford University Press for the Infectious Diseases Society of America. All rights reserved. For permissions, e-mail: journals.permissions@oup.com.
Chen, Ting; Jin, Yiying; Qiu, Xiaopeng; Chen, Xin
2015-03-01
Using laboratory experiments, the authors investigated the impact of dry-heat and moist-heat treatment processes on hazardous trace elements (As, Hg, Cd, Cr, and Pb) in food waste and explored their distribution patterns for three waste components: oil, aqueous, and solid components. The results indicated that an insignificant reduction of hazardous trace elements in heat-treated waste-0.61-14.29% after moist-heat treatment and 4.53-12.25% after dry-heat treatment-and a significant reduction in hazardous trace elements (except for Hg without external addition) after centrifugal dehydration (P < 0.5). Moreover, after heat treatment, over 90% of the hazardous trace elements in the waste were detected in the aqueous and solid components, whereas only a trace amount of hazardous trace elements was detected in the oil component (<0.01%). In addition, results indicated that heat treatment process did not significantly reduce the concentration of hazardous trace elements in food waste, but the separation process for solid and aqueous components, such as centrifugal dehydration, could reduce the risk considerably. Finally, combined with the separation technology for solid and liquid components, dry-heat treatment is superior to moist-heat treatment on the removal of external water-soluble ionic hazardous trace elements. An insignificant reduction of hazardous trace elements in heat-treated waste showed that heat treatment does not reduce trace elements contamination in food waste considerably, whereas the separation process for solid and aqueous components, such as centrifugal dehydration, could reduce the risk significantly. Moreover, combined with the separation technology for solid and liquid components, dry-heat treatment is superior to moist-heat treatment for the removal of external water-soluble ionic hazardous trace elements, by exploring distribution patterns of trace elements in three waste components: oil, aqueous, and solid components.
Hu, Li; Wang, Juan; Zhou, Xin; Xiong, Zehuan; Zhao, Jiajia; Yu, Ran; Huang, Fang; Zhang, Handong; Chen, Lili
2016-09-12
Prolonged healing and scar formation are two major challenges in the treatment of soft tissue trauma. Adipose mesenchymal stem cells (ASCs) play an important role in tissue regeneration, and recent studies have suggested that exosomes secreted by stem cells may contribute to paracrine signaling. In this study, we investigated the roles of ASCs-derived exosomes (ASCs-Exos) in cutaneous wound healing. We found that ASCs-Exos could be taken up and internalized by fibroblasts to stimulate cell migration, proliferation and collagen synthesis in a dose-dependent manner, with increased genes expression of N-cadherin, cyclin-1, PCNA and collagen I, III. In vivo tracing experiments demonstrated that ASCs-Exos can be recruited to soft tissue wound area in a mouse skin incision model and significantly accelerated cutaneous wound healing. Histological analysis showed increased collagen I and III production by systemic administration of exosomes in the early stage of wound healing, while in the late stage, exosomes might inhibit collagen expression to reduce scar formation. Collectively, our findings indicate that ASCs-Exos can facilitate cutaneous wound healing via optimizing the characteristics of fibroblasts. Our results provide a new perspective and therapeutic strategy for the use of ASCs-Exos in soft tissue repair.
Active learning of neuron morphology for accurate automated tracing of neurites
Gala, Rohan; Chapeton, Julio; Jitesh, Jayant; Bhavsar, Chintan; Stepanyants, Armen
2014-01-01
Automating the process of neurite tracing from light microscopy stacks of images is essential for large-scale or high-throughput quantitative studies of neural circuits. While the general layout of labeled neurites can be captured by many automated tracing algorithms, it is often not possible to differentiate reliably between the processes belonging to different cells. The reason is that some neurites in the stack may appear broken due to imperfect labeling, while others may appear fused due to the limited resolution of optical microscopy. Trained neuroanatomists routinely resolve such topological ambiguities during manual tracing tasks by combining information about distances between branches, branch orientations, intensities, calibers, tortuosities, colors, as well as the presence of spines or boutons. Likewise, to evaluate different topological scenarios automatically, we developed a machine learning approach that combines many of the above mentioned features. A specifically designed confidence measure was used to actively train the algorithm during user-assisted tracing procedure. Active learning significantly reduces the training time and makes it possible to obtain less than 1% generalization error rates by providing few training examples. To evaluate the overall performance of the algorithm a number of image stacks were reconstructed automatically, as well as manually by several trained users, making it possible to compare the automated traces to the baseline inter-user variability. Several geometrical and topological features of the traces were selected for the comparisons. These features include the total trace length, the total numbers of branch and terminal points, the affinity of corresponding traces, and the distances between corresponding branch and terminal points. Our results show that when the density of labeled neurites is sufficiently low, automated traces are not significantly different from manual reconstructions obtained by trained users. PMID:24904306
How Does the Sparse Memory “Engram” Neurons Encode the Memory of a Spatial–Temporal Event?
Guan, Ji-Song; Jiang, Jun; Xie, Hong; Liu, Kai-Yuan
2016-01-01
Episodic memory in human brain is not a fixed 2-D picture but a highly dynamic movie serial, integrating information at both the temporal and the spatial domains. Recent studies in neuroscience reveal that memory storage and recall are closely related to the activities in discrete memory engram (trace) neurons within the dentate gyrus region of hippocampus and the layer 2/3 of neocortex. More strikingly, optogenetic reactivation of those memory trace neurons is able to trigger the recall of naturally encoded memory. It is still unknown how the discrete memory traces encode and reactivate the memory. Considering a particular memory normally represents a natural event, which consists of information at both the temporal and spatial domains, it is unknown how the discrete trace neurons could reconstitute such enriched information in the brain. Furthermore, as the optogenetic-stimuli induced recall of memory did not depend on firing pattern of the memory traces, it is most likely that the spatial activation pattern, but not the temporal activation pattern of the discrete memory trace neurons encodes the memory in the brain. How does the neural circuit convert the activities in the spatial domain into the temporal domain to reconstitute memory of a natural event? By reviewing the literature, here we present how the memory engram (trace) neurons are selected and consolidated in the brain. Then, we will discuss the main challenges in the memory trace theory. In the end, we will provide a plausible model of memory trace cell network, underlying the conversion of neural activities between the spatial domain and the temporal domain. We will also discuss on how the activation of sparse memory trace neurons might trigger the replay of neural activities in specific temporal patterns. PMID:27601979
How Does the Sparse Memory "Engram" Neurons Encode the Memory of a Spatial-Temporal Event?
Guan, Ji-Song; Jiang, Jun; Xie, Hong; Liu, Kai-Yuan
2016-01-01
Episodic memory in human brain is not a fixed 2-D picture but a highly dynamic movie serial, integrating information at both the temporal and the spatial domains. Recent studies in neuroscience reveal that memory storage and recall are closely related to the activities in discrete memory engram (trace) neurons within the dentate gyrus region of hippocampus and the layer 2/3 of neocortex. More strikingly, optogenetic reactivation of those memory trace neurons is able to trigger the recall of naturally encoded memory. It is still unknown how the discrete memory traces encode and reactivate the memory. Considering a particular memory normally represents a natural event, which consists of information at both the temporal and spatial domains, it is unknown how the discrete trace neurons could reconstitute such enriched information in the brain. Furthermore, as the optogenetic-stimuli induced recall of memory did not depend on firing pattern of the memory traces, it is most likely that the spatial activation pattern, but not the temporal activation pattern of the discrete memory trace neurons encodes the memory in the brain. How does the neural circuit convert the activities in the spatial domain into the temporal domain to reconstitute memory of a natural event? By reviewing the literature, here we present how the memory engram (trace) neurons are selected and consolidated in the brain. Then, we will discuss the main challenges in the memory trace theory. In the end, we will provide a plausible model of memory trace cell network, underlying the conversion of neural activities between the spatial domain and the temporal domain. We will also discuss on how the activation of sparse memory trace neurons might trigger the replay of neural activities in specific temporal patterns.
Jia-En Zhang; Jiayu Yu; Ying Ouyang; Huaqin Xu
2014-01-01
Acid rain is one of the most serious ecological and environmental problems worldwide. This study investigated the impacts of simulated acid rain (SAR) upon leaching of trace metals and aluminum (Al) from a soil. Soil pot leaching experiments were performed to investigate the impacts of SAR at five different pH levels (or treatments) over a 34-day period upon the...
Internal Versus External DSLs for Trace Analysis: Extended Abstract
NASA Technical Reports Server (NTRS)
Barringer, Howard; Havelund, Klaus
2011-01-01
This tutorial explores the design and implementation issues arising in the development of domain-specific languages for trace analysis. It introduces the audience to the general concepts underlying such special-purpose languages building upon the authors' own experiences in developing both external domain specific languages and systems, such as EAGLE, HAWK, RULER and LOGSCOPE, and the more recent internal domain-specific language and system TRACECONTRACT within the SCALA language.
The Representation of Knowledge in Image Understanding.
1985-03-01
for memory traces? It’s been quite a few years since Lashley wrote his famous paper "Search for the Engram ." That paper pinpointed the fundamental...34 engram hunters," mostly empirical neurobiologists guided by their own often powerful working hypotheses, ensued and has continued that search with...progress. What is an Engram ? (Some Informal Considerations) Memory, viewed as traces of experience, is necessarily massively . distributed. As an example
Li, Guangjian; Huang, Yunchao; Liu, Yongjun; Guo, Lv; Zhou, Yongchun; Yang, Kun; Chen, Ying; Zhao, Guangqiang; Lei, Yujie
2012-10-01
China's Xuan Wei County in Yunnan Province have the world's highest incidence of lung cancer in nonsmoking women-20 times higher than the rest of China. Previous studies showed, this high lung cancer incidence may be associated with the silica particles embedded in the production combustion from the C1 coal. The aim of this study is to separate the silica particles from production combustion from the C1 bituminous coal in Xuan Wei County of Yunnan Province, and study in vitro toxicity of naturally occurring silica particles on BEAS-2B. ①Separating the silica particles from combustion products of C1 bituminous coal by physical method, observing the morphology by Scanning Electron Microscope, analysis elements by SEM-EDX, observed the single particle morphology by Transmission Electron Microscope, analyed its particle size distribution by Laser particle size analyzer, the surface area of silica particles were determined by BET nitrogen adsorption analysis; ②Cell viability of the experimental group (silica; naturally occurring), control group (silica; industrial produced and crystalline silica) was detected by assay used the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) method, and the reactive oxygen species (ROS), lactate dehydrogenase (LDH) were determined after 24 h-72 h exposed to these particles. ①The physical method can separate silica particles from production combustion from the C1 bituminous coal, which have different size, and from 30 nm to 120 nm particles accounted for 86.8%, different morphology, irregular surface area and containing trace of aluminum, calcium and iron and other elements; ②Under the same concentration, the experiment group have higher toxicity on BEAS-2B than control groups. Physical method can separate silica particles from production combustion from the C1 bituminous coal and not change the original morphology and containing trace; ②Naturally occurring silica nanoparticles have irregular morphology, surface area, and containing complex trace elements may has greater toxicity than the silica nanoparticle of industrial produced and crystalline silica.
Ray tracing: Experience at SRC
DOE Office of Scientific and Technical Information (OSTI.GOV)
Severson, M.
1996-09-01
SHADOW [B. Lai and F. Cerrina, Nucl. Instrum. Methods A {bold 246}, 337 (1986)] is the primary ray-tracing program used at SRC. Ray tracing provides a tremendous amount of information regarding beamline layout, mirror sizes, resolution, alignment tolerances, and beam size at various locations. It also provides a way to check the beamline design for errors. Two recent designs have been ray traced extensively: an undulator-based, 4-meter, normal-incidence monochromator (NIM) [R. Reininger, M.C. Severson, R.W.C. Hansen, W.R. Winter, M.A. Green, and W.S. Trzeciak, Rev. Sci. Instrum. {bold 66}, 2194 (1995)] and an undulator-based, plane-grating monochromator (PGM) [R. Reininger, S.L. Crossley,more » M.A. Lagergren, M.C. Severson, and R.W.C. Hansen, Nucl. Instrum. Methods A {bold 347}, 304 (1994)]. {copyright} {ital 1996 American Institute of Physics.}« less
Pardo, Tania; Martínez-Fernández, Domingo; Clemente, Rafael; Walker, David J; Bernal, M Pilar
2014-01-01
The applicability of a mature compost as a soil amendment to promote the growth of native species for the phytorestoration of a mine-affected soil from a semi-arid area (SE Spain), contaminated with trace elements (As, Cd, Cu, Mn, Pb and Zn), was evaluated in a 2-year field experiment. The effects of an inorganic fertiliser were also determined for comparison. Bituminaria bituminosa was the selected native plant since it is a leguminous species adapted to the particular local pedoclimatic conditions. Compost addition increased total organic-C concentrations in soil with respect to the control and fertiliser treatments, maintained elevated available P concentrations throughout the duration of the experiment and stimulated soil microbial biomass, while trace elements extractability in the soil was rather low due to the calcareous nature of the soil and almost unaltered in the different treatments. Tissue concentrations of P and K in B. bituminosa increased after the addition of compost, associated with growth stimulation. Leaf Cu concentration was also increased by the amendments, although overall the trace elements concentrations can be considered non-toxic. In addition, the spontaneous colonisation of the plots by a total of 29 species of 15 different families at the end of the experiment produced a greater vegetation cover, especially in plots amended with compost. Therefore, the use of compost as a soil amendment appears to be useful for the promotion of a vegetation cover and the phytostabilisation of moderately contaminated soils under semi-arid conditions.
An Efficient Format for Nearly Constant-Time Access to Arbitrary Time Intervals in Large Trace Files
Chan, Anthony; Gropp, William; Lusk, Ewing
2008-01-01
A powerful method to aid in understanding the performance of parallel applications uses log or trace files containing time-stamped events and states (pairs of events). These trace files can be very large, often hundreds or even thousands of megabytes. Because of the cost of accessing and displaying such files, other methods are often used that reduce the size of the tracefiles at the cost of sacrificing detail or other information. This paper describes a hierarchical trace file format that provides for display of an arbitrary time window in a time independent of the total size of the file and roughlymore » proportional to the number of events within the time window. This format eliminates the need to sacrifice data to achieve a smaller trace file size (since storage is inexpensive, it is necessary only to make efficient use of bandwidth to that storage). The format can be used to organize a trace file or to create a separate file of annotations that may be used with conventional trace files. We present an analysis of the time to access all of the events relevant to an interval of time and we describe experiments demonstrating the performance of this file format.« less
Both trace and delay conditioned eyeblink responding can be dissociated from outcome expectancy.
Weidemann, Gabrielle; Broderick, Joshua; Lovibond, Peter F; Mitchell, Christopher J
2012-01-01
Squire and colleagues have proposed that trace and delay eyeblink conditioning are fundamentally different kinds of learning: trace conditioning requires acquisition of a conscious declarative memory for the stimulus contingencies whereas delay conditioning does not. Declarative memory in trace conditioning is thought to generate conditioned responding through the activation of a conscious expectancy for when the unconditioned stimulus (US) is going to occur. Perruchet (1985) has previously shown that in a 50% partial reinforcement design it is possible to dissociate single cue delay eyeblink conditioning from conscious expectancy for the US by examining performance over runs of reinforced and nonreinforced trials. Clark, Manns, and Squire (2001) claim that this dissociation does not occur in trace eyeblink conditioning. In the present experiment we examined the Perruchet effect for short, moderate, and long trace intervals (600, 1000, and 1400 ms) and for the equivalent interstimulus intervals (ISIs) in a delay conditioning procedure. We found evidence for a dissociation of eyeblink CRs and US expectancy over runs regardless of whether there was a delay or a trace arrangement of cues. The reasons for the Perruchet effect are still unclear, but the present data suggest that it does not depend on a separate nondeclarative system of the type proposed by Squire and colleagues. (c) 2012 APA, all rights reserved.
Convective exosome-tracing microfluidics for analysis of cell-non-autonomous neurogenesis.
Oh, Hyun Jeong; Shin, Yoojin; Chung, Seok; Hwang, Do Won; Lee, Dong Soo
2017-01-01
The effective role of exosome delivering neurogenic microRNA (miRNA) enables to induce efficient differentiation process during neurogenesis. The microfludic system capable of visualizing the exosomal behavior such as secretion, migration, and uptake of individual exosomes can be used as a robust technique to understand the exosome-mediated change of cellular behavior. Here, we developed the exosome-tracing microfluidic system to visualize exosomal transport carrying the neurogenic miRNA from leading to neighboring cells, and found a new mode of exosome-mediated cell-non-autonomous neurogenesis. The miR-193a facilitated neurogenesis in F11 cells by blocking proliferation-related target genes. In addition to time-lapse live-cell imaging using microfluidics visualized the convective transport of exosomes from differentiated to undifferentiated cells. Individual exosomes containing miR-193a from differentiated donor cells were taken up by undifferentiated cells to lead them to neurogenesis. Induction of anti-miR-193a was sufficient to block neurogenesis in F11 cells. Inhibition of the exosomal production by manumycin-A and treatment of anti-miR-193a in the differentiated donor cells failed to induce neurogenesis in undifferentiated recipient cells. These findings indicate that exosomes of neural progenitors and neurogenic miRNA within these exosomes propagate cell-non-autonomous differentiation to neighboring progenitors, to delineate the roles of exosome mediating neurogenesis of population of homologous neural progenitor cells. Copyright © 2016 Elsevier Ltd. All rights reserved.
Lineage tracing of cells involved in atherosclerosis.
Albarrán-Juárez, Julián; Kaur, Harmandeep; Grimm, Myriam; Offermanns, Stefan; Wettschureck, Nina
2016-08-01
Despite the clinical importance of atherosclerosis, the origin of cells within atherosclerotic plaques is not fully understood. Due to the lack of a definitive lineage-tracing strategy, previous studies have provided controversial results about the origin of cells expressing smooth muscle and macrophage markers in atherosclerosis. We here aim to identify the origin of vascular smooth muscle (SM) cells and macrophages within atherosclerosis lesions. We combined a genetic fate mapping approach with single cell expression analysis in a murine model of atherosclerosis. We found that 16% of CD68-positive plaque macrophage-like cells were derived from mature SM cells and not from myeloid sources, whereas 31% of αSMA-positive smooth muscle-like cells in plaques were not SM-derived. Further analysis at the single cell level showed that SM-derived CD68(+) cells expressed higher levels of inflammatory markers such as cyclooxygenase 2 (Ptgs2, p = 0.02), and vascular cell adhesion molecule (Vcam1, p = 0.05), as well as increased mRNA levels of genes related to matrix synthesis such as Col1a2 (p = 0.01) and Fn1 (p = 0.04), than non SM-derived CD68(+) cells. These results demonstrate that smooth muscle cells within atherosclerotic lesions can switch to a macrophage-like phenotype characterized by higher expression of inflammatory and synthetic markers genes that may further contribute to plaque progression. Copyright © 2016 The Authors. Published by Elsevier Ireland Ltd.. All rights reserved.
Rao, S V Rama; Prakash, B; Raju, M V L N; Panda, A K; Kumari, R K; Reddy, E Pradeep Kumar
2016-08-01
Two experiments were conducted to study the effect of supplementing organic forms of zinc (Zn), selenium (Se) and chromium (Cr) on performance, anti-oxidant activities and immune responses in broiler chickens from 1 to 21 days of age, which were reared in cyclic heat-stressed condition under tropical summer in open-sided poultry house. A total of 200 (experiment I) and 450-day-old (experiment II) broiler male chicks (Cobb 400) were randomly distributed in stainless steel battery brooders (610 mm × 762 mm × 475 mm) at the rate of five birds per pen. A maize-soybean meal-based control diet (CD) containing recommended (Vencobb 400, Broiler Management Guide) concentrations of inorganic trace minerals and other nutrients was prepared. The CD was supplemented individually with organic form of selenium (Se, 0.30 mg/kg), chromium (Cr, 2 mg/kg) and zinc (Zn, 40 mg/kg) in experiment I. In experiment II, two concentrations of each Zn (20 and 40 mg/kg), Se (0.15 and 0.30 mg/kg) and Cr (1 and 2 mg/kg) were supplemented to the basal diet in 2 × 2 × 2 factorial design. A group without supplementing inorganic trace minerals was maintained as control group in both experiments. Each diet was allotted randomly to ten replicates in both experiments and fed ad libitum from 1 to 21 days of age. At 19th day of age, blood samples were collected for estimation of anti-oxidant and immune responses. Supplementation of Se, Cr and Zn increased (P < 0.05) body mass gain (BMG) and feed intake compared to those fed the CD in experiment I. The feed efficiency (FE) in Cr-fed group was higher (P < 0.05) compared to the CD-fed group. Se or Cr supplementation reduced lipid peroxidation (LP) compared to broilers fed the CD. In experiment II, BMG was not affected (P > 0.05) by the interaction between levels of Zn, Se and Cr in broiler diet. The FE improved (P < 0.05) with supplementation of the trace minerals tested at both concentrations except in group fed 40 mg Zn, 0.5 mg Se and 1 mg Cr/kg. Reduction in lipid peroxidation (LP, P < 0.05) and increased (P < 0.05) activity of superoxide dismutase were observed in broiler fed organic Zn, Se and Cr compared to the CD-fed group. The dietary concentrations of Zn, Se and Cr did not influence (P > 0.05) the immune responses (Newcastle disease titre and cell-mediated immune response to phytohaemagglutinin-P) in both the experiments. Based on the results, it is concluded that supplementation of organic form of Se, Cr and Zn (0.30, 2 and 40 mg/kg, respectively) either alone or in combination significantly improved performance and anti-oxidant responses (reduced LP and increased superoxide dismutase) in commercial broiler chicks (21 days of age) reared in cyclic heat stress conditions in open-sided poultry house during summer.
Visualizing speciation in artificial cichlid fish.
Clement, Ross
2006-01-01
The Cichlid Speciation Project (CSP) is an ALife simulation system for investigating open problems in the speciation of African cichlid fish. The CSP can be used to perform a wide range of experiments that show that speciation is a natural consequence of certain biological systems. A visualization system capable of extracting the history of speciation from low-level trace data and creating a phylogenetic tree has been implemented. Unlike previous approaches, this visualization system presents a concrete trace of speciation, rather than a summary of low-level information from which the viewer can make subjective decisions on how speciation progressed. The phylogenetic trees are a more objective visualization of speciation, and enable automated collection and summarization of the results of experiments. The visualization system is used to create a phylogenetic tree from an experiment that models sympatric speciation.
Cell-adhesion molecules in memory formation.
Schmidt, R
1995-01-23
After learning events the CNS of higher organisms selects, which acquired informations are permanently stored as a memory trace. This period of memory consolidation is susceptible to interference by biochemical inhibitors of transcription and translation. Ependymin is a specific CNS glycoprotein functionally involved in memory consolidation in goldfish: after active shock-avoidance conditioning ependymin mRNA is rapidly induced in meningeal fibroblasts followed by enhanced synthesis and secretion of several closely related forms of the protein. Intracranial injections of anti-ependymin antisera or antisense oligodeoxynucleotides interfere specifically with memory consolidation, indicating that only de novo synthesized ependymin molecules are involved. Ependymin is capable of directing the growth of central axons in vitro and participates in neuronal regeneration in situ, presumably by its HNK-1 cell-adhesion epitope. Experiments reviewed in this article suggest a model that involves two regulation mechanisms for the function of ependymin in behavioural plasticity: while hormones appear to determine, how much of this cell adhesion molecule is synthesized after learning, local changes of metal cation concentrations in the micro-environment of activated neurons may polymerize ependymin at those synapses, that have to be consolidated to improve their efficacy for future use.
Metalizing Solar Cells by Selective Electroplating
NASA Technical Reports Server (NTRS)
Dutta, S.; Palaschak, P. A.
1986-01-01
Contact patterns traced by laser scanning. Conductor paths deposited on silicon solar-cell wafers by laser irradiation followed by electroplating. Laser-assisted metalization technique offers better resolution and lower contact resistance than does conventional metalization by screen printing. At the same time, less expensive than metalization with masks and photolithography.
NASA Astrophysics Data System (ADS)
Lloyd, Charlotte; Michaelides, Katerina; Evershed, Richard; Chadwick, David; Dungait, Jennifer
2010-05-01
We explore the use of organic biomarkers as tracers for different components of livestock-derived organic matter (LD-OM) at two different spatial scales. We conducted six small-scale rainfall simulation experiments on a 30 × 30 × 30 cm soil lysimeter, following an application of bovine slurry at a rate of 5 l m-2. Throughout the experiment timed samples of leachate from the base of the lysimeter were collected, then soil cores were taken following the rainfall simulation. These samples were analysed in order to identify the most suitable biomarkers to trace dissolved and sediment-bound LD-OM respectively. The results showed that ammonium was an important tracer compound for dissolved LD-OM, along with other key low molecular weight compounds such as carbohydrates and amino acids. Analysis of the soil cores confirmed that compounds 5-β sigmastanol and 5-β epistigmastanol (5-β stanols) could be used very effectively to trace the sediment-bound and colloidal component of LD-OM. These specific organic compounds, which are identifiable by GC/MS analysis, only occur due to biohydrogenation of plant sterols in a ruminant gut, providing a unique opportunity to trace bovine faecal matter via sediment pathways. These tracers were then applied to a larger 3-D hillslope system by using University of Bristol's TRACE (Test Rig for Advancing Connectivity Experiments) facility. TRACE is a large-scale dual axis soil-slope measuring 6 m long × 2.5 m wide × 0.3 m deep accompanied by a 6-nozzle rainfall simulator. In these experiments slurry was only applied to the top 1 m section of the hillslope, in order to trace how the LD-OM was transported in the soil system. The slope allows the collection of leachate from the soil surface, from lateral through-flow and infiltrated water which reached the soil base (indicating deeper pathways). This enabled the distinction between LD-OM transported via different hydrological pathways. Soil cores were also taken across the soil surface and analysed for 5-β stanols, this allowed the spatial distribution of LD-OM to be determined following the rainfall event. The results showed that not only is LD-OM transported on the surface of the hillslope via overland flow, but the dissolved component infiltrates through the soil profile and is transported via deeper hydrological flowpaths. 5-β stanol analysis showed that soil erosion processes were extremely important, as LD-OM was found downslope of the application area and in eroded material lost from the base of the experimental hillslope. These experiments provided new insights into how LD-OM interacts with the soil-water system and allows quantification of the contamination risk posed. This is important as 90 million tonnes of LD-OM is applied to land annually in the UK. It is well known that there is a potential for contamination of water courses by nitrate, ammonium and other faecal-derived pollutants such as E. Coli through runoff from treated land. Pollution from LD-OM has now been shown to extend to the contamination of subsurface pathways and potentially groundwater resources.
Liu, Yina; Thornton, Daniel C O; Bianchi, Thomas S; Arnold, William A; Shields, Michael R; Chen, Jie; Yvon-Lewis, Shari A
2015-03-17
Brominated very short-lived substances (BrVSLS), such as bromoform, are important trace gases for stratospheric ozone chemistry. These naturally derived trace gases are formed via bromoperoxidase-mediated halogenation of dissolved organic matter (DOM) in seawater. Information on DOM type in relation to the observed BrVSLS concentrations in seawater, however, is scarce. We examined the sensitivity of BrVSLS production in relation to the presence of specific DOM moieties. A total of 28 model DOM compounds in artificial seawater were treated with vanadium bromoperoxidase (V-BrPO). Our results show a clear dependence of BrVSLS production on DOM type. In general, molecules that comprise a large fraction of the bulk DOM pool did not noticeably affect BrVSLS production. Only specific cell metabolites and humic acid appeared to significantly enhance BrVSLS production. Amino acids and lignin phenols suppressed enzyme-mediated BrVSLS production and may instead have formed halogenated nonvolatile molecules. Dibromomethane production was not observed in any experiments, suggesting it is not produced by the same pathway as the other BrVSLS. Our results suggest that regional differences in DOM composition may explain the observed BrVSLS concentration variability in the global ocean. Ultimately, BrVSLS production and concentrations are likely affected by DOM composition, reactivity, and cycling in the ocean.
Spectral radiation analyses of the GOES solar illuminated hexagonal cell scan mirror back
NASA Technical Reports Server (NTRS)
Fantano, Louis G.
1993-01-01
A ray tracing analytical tool has been developed for the simulation of spectral radiation exchange in complex systems. Algorithms are used to account for heat source spectral energy, surface directional radiation properties, and surface spectral absorptivity properties. This tool has been used to calculate the effective solar absorptivity of the geostationary operational environmental satellites (GOES) scan mirror in the calibration position. The development and design of Sounder and Imager instruments on board GOES is reviewed and the problem of calculating the effective solar absorptivity associated with the GOES hexagonal cell configuration is presented. The analytical methodology based on the Monte Carlo ray tracing technique is described and results are presented and verified by experimental measurements for selected solar incidence angles.
2007-02-27
intervals and amplitude of sIPSCs, in baseline conditions and during !-Methyl- 5- HT application (same cell as in the top trace). (C) Up trace: effects of...Methyl-5- HT (15 #M) on sIPSCs recorded from a BLA pyramidal cell of a stressed rat (holding potential is -70 mV); the effect of !-Methyl-5- HT (15...dysfunctions in rats. Chapter 2–Figure 1— Effect of 5- HT on sIPSCs in the basolateral amygdala. Chapter 2-Figure 2— The 5-HT2 receptor agonist !-Methyl
NASA Astrophysics Data System (ADS)
Washenfelder, R. A.; Attwood, A. R.; Flores, J. M.; Rudich, Y.; Brown, S. S.
2015-09-01
Formaldehyde (CH2O) is the most abundant aldehyde in the atmosphere, and strongly affects photochemistry through its photolysis. We describe simultaneous measurements of CH2O and nitrogen dioxide (NO2) using broadband cavity enhanced spectroscopy in the ultraviolet spectral region. The light source consists of a continuous-wave diode laser focused into a Xenon bulb to produce a plasma that emits high-intensity, broadband light. The plasma discharge is optically filtered and coupled into a 1 m optical cavity. The reflectivity of the cavity mirrors is 0.99933 ± 0.00003 (670 ppm loss) at 338 nm, as determined from the known Rayleigh scattering of He and zero air. This mirror reflectivity corresponds to an effective path length of 1.49 km within the 1 m cell. We measure the cavity output over the 315-350 nm spectral region using a grating monochromator and charge-coupled device (CCD) array detector. We use published reference spectra with spectral fitting software to simultaneously retrieve CH2O and NO2 concentrations. Independent measurements of NO2 standard additions by broadband cavity enhanced spectroscopy and cavity ringdown spectroscopy agree within 2 % (slope for linear fit = 0.98 ± 0.03 with r2 = 0.998). Standard additions of CH2O measured by broadband cavity enhanced spectroscopy and calculated based on flow dilution are also well-correlated, with r2 = 0.9998. During constant, mixed additions of NO2 and CH2O, the 30 s measurement precisions (1σ) of the current configuration were 140 and 210 pptv, respectively. The current 1-min detection limit for extinction measurements at 315-350 nm provides sufficient sensitivity for measurement of trace gases in laboratory experiments and ground-based field experiments. Additionally, the instrument provides highly accurate, spectroscopically-based trace gas detection that may complement higher precision techniques based on non-absolute detection methods. In addition to trace gases, this approach will be appropriate for measurements of aerosol extinction in ambient air, and this spectral region is important for characterizing the strong ultraviolet absorption by brown carbon aerosol.
Davis, Shannon W; Keisler, Jessica L; Pérez-Millán, María I; Schade, Vanessa; Camper, Sally A
2016-04-01
Mutations in PROP1, the most common known cause of combined pituitary hormone deficiency in humans, can result in the progressive loss of all hormones of the pituitary anterior lobe. In mice, Prop1 mutations result in the failure to initiate transcription of Pou1f1 (also known as Pit1) and lack somatotropins, lactotropins, and thyrotropins. The basis for this species difference is unknown. We hypothesized that Prop1 is expressed in a progenitor cell that can develop into all anterior lobe cell types, and not just the somatotropes, thyrotropes, and lactotropes, which are collectively known as the PIT1 lineage. To test this idea, we produced a transgenic Prop1-cre mouse line and conducted lineage-tracing experiments of Prop1-expressing cells. The results reveal that all hormone-secreting cell types of both the anterior and intermediate lobes are descended from Prop1-expressing progenitors. The Prop1-cre mice also provide a valuable genetic reagent with a unique spatial and temporal expression for generating tissue-specific gene rearrangements early in pituitary gland development. We also determined that the minimal essential sequences for reliable Prop1 expression lie within 10 kilobases of the mouse gene and demonstrated that human PROP1 can substitute functionally for mouse Prop1. These studies enhance our understanding of the pathophysiology of disease in patients with PROP1 mutations.
High sensitivity detection of trace gases at atmospheric pressure using tunable diode lasers
NASA Technical Reports Server (NTRS)
Reid, J.; Sinclair, R. L.; Grant, W. B.; Menzies, R. T.
1985-01-01
A detailed study of the detection of trace gases at atmospheric pressure using tunable diode lasers is described. The influence of multipass cells, retroreflectors and topographical targets is examined. The minimum detectable infrared absorption ranges from 0.1 percent for a pathlength of 1.2 km to 0.01 percent over short pathlengths. The factors which limit this sensitivity are discussed, and the techniques are illustrated by monitoring atmospehric CO2 and CH4.
Photoacoustic Spectroscopy with Quantum Cascade Lasers for Trace Gas Detection
Elia, Angela; Di Franco, Cinzia; Lugarà, Pietro Mario; Scamarcio, Gaetano
2006-01-01
Various applications, such as pollution monitoring, toxic-gas detection, non invasive medical diagnostics and industrial process control, require sensitive and selective detection of gas traces with concentrations in the parts in 109 (ppb) and sub-ppb range. The recent development of quantum-cascade lasers (QCLs) has given a new aspect to infrared laser-based trace gas sensors. In particular, single mode distributed feedback QCLs are attractive spectroscopic sources because of their excellent properties in terms of narrow linewidth, average power and room temperature operation. In combination with these laser sources, photoacoustic spectroscopy offers the advantage of high sensitivity and selectivity, compact sensor platform, fast time-response and user friendly operation. This paper reports recent developments on quantum cascade laser-based photoacoustic spectroscopy for trace gas detection. In particular, different applications of a photoacoustic trace gas sensor employing a longitudinal resonant cell with a detection limit on the order of hundred ppb of ozone and ammonia are discussed. We also report two QC laser-based photoacoustic sensors for the detection of nitric oxide, for environmental pollution monitoring and medical diagnostics, and hexamethyldisilazane, for applications in semiconductor manufacturing process.
Lipids of Pseudomonas aeruginosa Cells Grown on Hydrocarbons and on Trypticase Soy Broth1
Edmonds, Paul; Cooney, J. J.
1969-01-01
Lipids were extracted from cells of Pseudomonas aeruginosa grown on a pure hydrocarbon (tridecane), mixed hydrocarbons (JP-4 jet fuel), and on Trypticase Soy Broth. Total lipids produced from each substrate represented from 7.1 to 8.2% of cellular dry weight, of which 5.0 to 6.4% were obtained before cellular hydrolysis (free lipids) and 1.7 to 2.0% were extracted after cellular hydrolysis (bound lipids). Free lipids from cells grown on each medium were separated into four fractions by thin-layer chromatography. All fractions were present in cells from each type of medium, and the “neutral fraction” constituted the largest fraction. The fatty acid composition of free lipids was determined by gas-liquid chromatography. Cells grown on each medium contained saturated and unsaturated C14 to C20 fatty acids. Trace amounts of C13 fatty acids were found in tridecane-grown cells. Saturated C16 and C18 were the major acids present in all cells. Quantitative differences were found in fatty acids produced on the three media, but specific correlations between substrate carbon sources and fatty acid content of cells were not evident. Tridecane-grown cells contained only traces of C13 acid and small amounts of C15 and C17 acids, suggesting that the organism's fatty acids were derived from de novo synthesis rather than by direct incorporation of the hydrocarbon. PMID:4976464
Chen, Peng; Wang, Zhuyuan; Zong, Shenfei; Zhu, Dan; Chen, Hui; Zhang, Yizhi; Wu, Lei; Cui, Yiping
2016-01-15
We fabricate a multifunctional nanocarrier based on multi-walled carbon nanotubes (MWCNTs) decorated with gold/silver core-shell nanoparticles (Au@Ag NPs) and fluorescein isothiocyanate (FITC) for tracking the intracellular drug release process. In the demonstrated nanocarrier, the Au@Ag NPs adsorbed on the surface of MWCNTs were labeled with the pH-dependent SERS reporter 4-Mercaptobenzoic acid (4MBA) for SERS based pH sensing. FITC was conjugated on MWCNTs to provide fluorescence signal for tracing the MWCNTs. Fluorescent doxorubicin (DOX) was used as the model drug which can be loaded onto MWCNTs via π-π stacking and released from the MWCNTs under acidic condition. By detecting the SERS spectrum of 4MBA, the pH value around the nanocarrier could be monitored. Besides, by tracing the fluorescence of FITC and DOX, we can also investigate the drug release process in cells. Experimental results show that the proposed nanocarrier retained a well pH-sensitive performance in living cells, and the DOX detached from MWCNTs inside the lysosomes and entered into the cytoplasm with the MWCNTs being left in lysosomes. To further investigate the drug release dynamics, 2-D color-gradient pH mapping were plotted, which were calculated from the SERS spectra of 4MBA. The detailed release process and carrier distribution have been recorded as environmental pH changes during cell endocytosis. Furthermore, we also confirmed that the proposed nanocarrier has a good biocompatibility. It indicates that the designed nanocarrier have a great potential in intraceable drug delivery, cancer cells imaging and pH monitoring. Copyright © 2015 Elsevier B.V. All rights reserved.
Li, J; Xiong, K H; Li, Y Q; Kaneko, T; Mizuno, N
2000-06-01
Neurons of the mesencephalic trigeminal nucleus (MTN) are considered to be homologous to mechanosensitive neurons in the sensory ganglia. The sites of origin of serotonin (5HT)-immunoreactive axons on neuronal cell bodies in the MTN were studied in the rat by combining immunofluorescence histochemical techniques with retrograde tracing of Fluoro-Gold (FG) and anterograde tracing of biotin-conjugated dextran amine (BDA). The tracing studies, which were combined with multiple-labeling immunohistochemistry and confocal microscopy, indicated that 5HT-immunoreactive axon terminals on the cell bodies of MTN neurons originated from the medullary raphe nuclei, such as the nucleus raphes magmus (RMg), alpha part of the nucleus reticularis gigantocellularis (GiA) and nucleus raphes obscurus (ROb), as well as from the mesopontine raphe nuclei, such as the nucleus raphes dorsalis (DR), nucleus raphes pontis (PnR) and nucleus raphes medianus (MnR); mainly from the RMg, GiA and DR, and additionally from the ROb, PnR and MnR. The cell bodies in close apposition to 5HT-immunoreactive axon terminals were found through the whole rostrocaudal extent of the MTN. Electron microscopically a number of axon terminals that were labeled with BDA injected into the raphe nuclei were confirmed to be in asymmetric synaptic contact with the cell bodies of MTN neurons. It was also indicated that substance P existed in some of the 5HT-containing axosomatic terminals arising from the ROb, RMg and GiA. The present results indicated that proprioceptive sensory signals from the muscle spindles or periodontal ligament might be modulated at the level of the primary afferent cell bodies in the MTN by 5HT-containing axons from the mesopontine and medullary raphe nuclei including the GiA.
Histological Methods for ex vivo Axon Tracing: A Systematic Review
Heilingoetter, Cassandra L.; Jensen, Matthew B.
2016-01-01
Objectives Axon tracers provide crucial insight into the development, connectivity, and function of neural pathways. A tracer can be characterized as a substance that allows for the visualization of a neuronal pathway. Axon tracers have previously been used exclusively with in vivo studies; however, newer methods of axon tracing can be applied to ex vivo studies. Ex vivo studies involve the examination of cells or tissues retrieved from an organism. These post mortem methods of axon tracing offer several advantages, such as reaching inaccessible tissues and avoiding survival surgeries. Methods In order to evaluate the quality of the ex vivo tracing methods, we performed a systematic review of various experimental and comparison studies to discern the optimal method of axon tracing. Results The most prominent methods for ex vivo tracing involve enzymatic techniques or various dyes. It appears that there are a variety of techniques and conditions that tend to give better fluorescent character, clarity, and distance traveled in the neuronal pathway. We found direct comparison studies that looked at variables such as the type of tracer, time required, effect of temperature, and presence of calcium, however, there are other variables that have not been compared directly. Discussion We conclude there are a variety of promising tracing methods available depending on the experimental goals of the researcher, however, more direct comparison studies are needed to affirm the optimal method. PMID:27098542
Histological methods for ex vivo axon tracing: A systematic review.
Heilingoetter, Cassandra L; Jensen, Matthew B
2016-07-01
Axon tracers provide crucial insight into the development, connectivity, and function of neural pathways. A tracer can be characterized as a substance that allows for the visualization of a neuronal pathway. Axon tracers have previously been used exclusively with in vivo studies; however, newer methods of axon tracing can be applied to ex vivo studies. Ex vivo studies involve the examination of cells or tissues retrieved from an organism. These post mortem methods of axon tracing offer several advantages, such as reaching inaccessible tissues and avoiding survival surgeries. In order to evaluate the quality of the ex vivo tracing methods, we performed a systematic review of various experimental and comparison studies to discern the optimal method of axon tracing. The most prominent methods for ex vivo tracing involve enzymatic techniques or various dyes. It appears that there are a variety of techniques and conditions that tend to give better fluorescent character, clarity, and distance traveled in the neuronal pathway. We found direct comparison studies that looked at variables such as the type of tracer, time required, effect of temperature, and presence of calcium, however, there are other variables that have not been compared directly. We conclude there are a variety of promising tracing methods available depending on the experimental goals of the researcher, however, more direct comparison studies are needed to affirm the optimal method.
ERIC Educational Resources Information Center
White, Peter A.
2012-01-01
Forces are experienced in actions on objects. The mechanoreceptor system is stimulated by proximal forces in interactions with objects, and experiences of force occur in a context of information yielded by other sensory modalities, principally vision. These experiences are registered and stored as episodic traces in the brain. These stored…
Fluorescent proteins such as eGFP lead to catalytic oxidative stress in cells.
Ganini, Douglas; Leinisch, Fabian; Kumar, Ashutosh; Jiang, JinJie; Tokar, Erik J; Malone, Christine C; Petrovich, Robert M; Mason, Ronald P
2017-08-01
Fluorescent proteins are an important tool that has become omnipresent in life sciences research. They are frequently used for localization of proteins and monitoring of cells [1,2]. Green fluorescent protein (GFP) was the first and has been the most used fluorescent protein. Enhanced GFP (eGFP) was optimized from wild-type GFP for increased fluorescence yield and improved expression in mammalian systems [3]. Many GFP-like fluorescent proteins have been discovered, optimized or created, such as the red fluorescent protein TagRFP [4]. Fluorescent proteins are expressed colorless and immature and, for eGFP, the conversion to the fluorescent form, mature, is known to produce one equivalent of hydrogen peroxide (H 2 O 2 ) per molecule of chromophore [5,6]. Even though it has been proposed that this process is non-catalytic and generates nontoxic levels of H 2 O 2 [6], this study investigates the role of fluorescent proteins in generating free radicals and inducing oxidative stress in biological systems. Immature eGFP and TagRFP catalytically generate the free radical superoxide anion (O 2 •- ) and H 2 O 2 in the presence of NADH. Generation of the free radical O 2 •- and H 2 O 2 by eGFP in the presence of NADH affects the gene expression of cells. Many biological pathways are altered, such as a decrease in HIF1α stabilization and activity. The biological pathways altered by eGFP are known to be implicated in the pathophysiology of many diseases associated with oxidative stress; therefore, it is critical that such experiments using fluorescent proteins are validated with alternative methodologies and the results are carefully interpreted. Since cells inevitably experience oxidative stress when fluorescent proteins are expressed, the use of this tool for cell labeling and in vivo cell tracing also requires validation using alternative methodologies. Published by Elsevier B.V.
Guo, Liang; Lichten, Louis A.; Ryu, Moon-Suhn; Liuzzi, Juan P.; Wang, Fudi; Cousins, Robert J.
2010-01-01
The exocrine pancreas plays an important role in endogenous zinc loss by regulating excretion into the intestinal tract and hence influences the dietary zinc requirement. The present experiments show that the zinc transporter ZnT2 (Slc30a2) is localized to the zymogen granules and that dietary zinc restriction in mice decreased the zinc concentration of zymogen granules and ZnT2 expression. Excess zinc given orally increased ZnT2 expression and was associated with increased pancreatic zinc accumulation. Rat AR42J acinar cells when induced into a secretory phenotype, using the glucocorticoid analog dexamethasone (DEX), exhibited increased ZnT2 expression and labile zinc as measured with a fluorophore. DEX administrated to mice also induced ZnT2 expression that accompanied a reduction of the pancreatic zinc content. ZnT2 promoter analyses identified elements required for responsiveness to zinc and DEX. Zinc regulation was traced to a MRE located downstream from the ZnT2 transcription start site. Responsiveness to DEX is produced by two upstream STAT5 binding sites that require the glucocorticoid receptor for activation. ZnT2 knockdown in the AR42J cells using siRNA resulted in increased cytoplasmic zinc and decreased zymogen granule zinc that further demonstrated that ZnT2 may mediate the sequestration of zinc into zymogen granules. We conclude, based upon experiments with intact mice and pancreatic acinar cells in culture, that ZnT2 participates in zinc transport into pancreatic zymogen granules through a glucocorticoid pathway requiring glucocorticoid receptor and STAT5, and zinc-regulated signaling pathways requiring MTF-1. The ZnT2 transporter appears to function in a physiologically responsive manner involving entero-pancreatic zinc trafficking. PMID:20133611
Openness to Experience as a Basic Dimension of Personality.
ERIC Educational Resources Information Center
McCrae, Robert R.
This paper opens by describing research since 1975 (McCrae and Costa) on a set of related traits that identified as aspects of Openness to Experience. The historic roots of the concept of Openness to Experience are traced. Data are provided on the convergent and discriminant validity of the six Revised NEO-Personality Inventory facets of Fantasy,…
Rei, Mariana; Tavares, Sara; Pinto, Pedro; Machado, Ana P; Monteiro, Sofia; Costa, Antónia; Costa-Santos, Cristina; Bernardes, João; Ayres-De-Campos, Diogo
2016-10-01
Visual analysis of cardiotocographic (CTG) tracings has been shown to be prone to poor intra- and interobserver agreement when several interpretation guidelines are used, and this may have an important impact on the technology's performance. The aim of this study was to evaluate agreement in CTG interpretation using the new 2015 FIGO guidelines on intrapartum fetal monitoring. A pre-existing database of intrapartum CTG tracings was used to sequentially select 151 cases acquired with a fetal electrode, with duration exceeding 60minutes, and signal loss less than 15%. These tracings were presented to six clinicians, three with more than 5 years' experience in the labor ward, and three with 5 or less years' experience. Observers were asked to evaluate tracings independently, to assess basic CTG features: baseline, variability, accelerations, decelerations, sinusoidal pattern, tachysystole, and to classify each tracing as normal, suspicious or pathologic, according to the 2015 FIGO guidelines on intrapartum fetal monitoring. Agreement between observers was evaluated using the proportions of agreement (Pa), with 95% confidence intervals (95%CI). A good interobserver agreement was found in the evaluation of most CTG features, but not bradycardia, reduced variability, saltatory pattern, absence of accelerations and absence of decelerations. For baseline classification Pa was 0.85 [0.82-0.90], for variability 0.82 [0.78-0.85], for accelerations 0.72 [0.68-0.75], for tachysystole 0.77 [0.74-0.81], for decelerations 0.92 [0.90-0.95], for variable decelerations 0.62 [0.58-0.65], for late decelerations 0.63 [0.59-0.66], for repetitive decelerations 0.73 [0.69-0.78], and for prolonged decelerations 0.81 [0.77-0.85]. For overall CTG classification, Pa were 0.60 [0.56-0.64], for classification as normal 0.67 [0.61-0.72], for suspicious 0.54 [0.48-0.60] and for pathologic 0.59 [0.51-0.66]. No differences in agreement according to the level of expertise were observed, except in the identification of accelerations, where it was better in the more experienced group. A good interobserver agreement was found in evaluation of most CTG features and in overall tracing classification. Results were better than those reported in previous studies evaluating agreement in overall tracing classification. Observer experience did not appear to play a role in agreement. Copyright © 2016 Elsevier Ireland Ltd. All rights reserved.
Star-shaped feeding traces produced by echiuran worms on the deep-sea floor of the Bay of Bengal
NASA Astrophysics Data System (ADS)
Ohta, Suguru
1984-12-01
Many star-shaped foraging traces were observed in bottom photographs of the deep-sea floor taken in the Bay of Bengal between the depths of 5025 and 2635 m. They were classified into 10 types according to their dimensions, aspect ratios (length/width) of their spokes, features of the central structure, and possible production mechanisms. The proboscis of a deep-sea bonellid echiuran worm was photographed at a depth of 2635 m in the act of producing one of the star-shaped foraging traces. On the basis of photographic observations and observations of shallow-water forms, several types of the feeding traces can be ascribed to the foraging of deep-sea echiuran worms on surface detritus. At least four types of the star-shaped trace are probably produced by deep-sea bonellid worms, and a linear correlation could be found between the aspect ratios of the spokes and maximum number of spokes around the central hole. A geometrical model experiment stimulating the feeding behavior of a bonellid worm suggested simple behavioral principles which afford maximum utilization of the surface area around a central hole with least expenditure of energy. The prediction of the maximum number of spokes for a given aspect of spokes by the model experiment agreed well with those observed, both utilizing about 76% of the fresh sediment surface within the span of the probiscis around a central hole. This efficient feeding pattern may have adaptive value in deep-sea environments such as the central part of the Bay of Bengal, where energy input is limited.
Mid-Frequency Reverberation Measurements with Full Companion Environmental Support
2014-12-30
acoustic modeling is based on measured stratification and observed wave amplitudes on the New Jersey shelf during the SWARM experiment.3 Ray tracing is...wave model then gives quantitative results for the clutter. 2. Swarm NLIW model and ray tracing Nonlinear internal waves are very common on the...receiver in order to give quantitative clutter to reverberation. To picture the mechanism, a set of rays was launched from a source at range zero and
Visual Information-Processing in the Perception of Features and Objects
1989-01-05
or nodes in a semantic memory network, whereas recall and recognition depend on separate episodic memory traces. In our experiment, we used the same...problem for the account in terms of the separation of episodic from semantic memory , since no pre- existing representations of our line patterns were... semantic memory : amnesic patients were thought to have lost the ability to lay down (or retrieve) episodic traces of autobiographical events, but had
Trinity Bay Study: Dye tracing experiments
NASA Technical Reports Server (NTRS)
Ward, G. H., Jr.
1972-01-01
An analysis of the heat balance and temperature distribution within Trinity Bay near Galveston, Texas is presented. The effects of tidal currents, wind driven circulations, and large volume inflows are examined. Emphasis is placed on the effects of turbulent diffusion and local shears in currents. The technique of dye tracing to determine the parameters characterizing dispersion is described. Aerial photographs and maps are provided to show the flow conditions existing at different times and seasons.
Models and Measurements for Multi-Layer Displays
2006-07-26
measurements. The observed statistical variation in the data results from laser speckle. No systematic uncertainties, which are expected to be less...difference metric. There are also some powerful statistical techniques to deal with this type of experiment, although it would take a lot of time to...hTraceWidth,vTraceWidth] in 10s of micrometers % Transitor sixe is vector : [hTransistorSize,vTransistorSize] in 10s of micrometers %Image is plotted if
DOE Office of Scientific and Technical Information (OSTI.GOV)
Pastorok, R.; Schoof, R.; LaTier, A.
1995-12-31
At mining and smelting sites, the bioavailability of waste-related trace elements to terrestrial wildlife is limited by mineralogy of the waste material and the geochemistry of the waste-soil mixture. For example, encapsulation of trace elements in inert mineral matrices limits the assimilation of particle-associated trace elements that are ingested by wildlife. The bioavailability of arsenic, cadmium, copper, lead, silver, and zinc at mining and smelting sites in Oklahoma and Montana was evaluated based on analysis of waste material, soil chemistry, and concentrations of trace elements in whole-body samples of key food web species. Concentrations of trace elements were generally elevatedmore » relative to reference area values for selected species of vegetation, insects, spiders, and small mammals. Soil-to-tissue bioconcentration factors derived from field data at these sites were generally low (< 1), with the exception of cadmium in vegetation. For all of the trace elements evaluated, wildlife exposure models indicate that the potential for transfer of contaminants to wildlife species of public concern and high trophic-level predators is limited. Moreover, laboratory feeding experiments conducted with cadmium and lead indicate that the assimilation of waste-related trace elements by mammals is relatively low (24--47 percent for lead in blood and bone; 22--44 percent for cadmium in kidney). The relatively low bioavailability of trace elements at mining and smelting sites should be considered when estimating exposure of ecological receptors and when deriving soil cleanup criteria based on measured or modeled ecological risk.« less
Trivedi, Mehul A; Coover, Gary D
2006-04-03
Pavlovian delay conditioning, in which a conditioned stimulus (CS) and unconditioned stimulus (US) co-terminate, is thought to reflect non-declarative memory. In contrast, trace conditioning, in which the CS and US are temporally separate, is thought to reflect declarative memory. Hippocampal lesions impair acquisition and expression of trace conditioning measured by the conditioned freezing and eyeblink responses, while having little effect on the acquisition of delay conditioning. Recent evidence suggests that lesions of the ventral hippocampus (VH) impair conditioned fear under conditions in which dorsal hippocampal (DH) lesions have little effect. In the present study, we examined the time-course of fear expression after delay and trace conditioning using the fear-potentiated startle (FPS) reflex, and the effects of pre- and post-training lesions to the VH and DH on trace-conditioned FPS. We found that both delay- and trace-conditioned rats displayed significant FPS near the end of the CS relative to the unpaired control group. In contrast, trace-conditioned rats displayed significant FPS throughout the duration of the trace interval, whereas FPS decayed rapidly to baseline after CS offset in delay-conditioned rats. In experiment 2, both DH and VH lesions were found to significantly reduce the overall magnitude of FPS compared to the control group, however, no differences were found between the DH and VH groups. These findings support a role for both the DH and VH in trace fear conditioning, and suggest that the greater effect of VH lesions on conditioned fear might be specific to certain measures of fear.
NASA Astrophysics Data System (ADS)
Tan, Y. S.; Ang, A.; Gomes, H. R.; Goes, J. I.
2016-12-01
Noctiluca scintillans, a species of mixotrophic dinoflagellate, has been proliferating in the Arabian Sea for the past 20 years in winter when convective mixing brings up nutrient-rich waters from deeper depths. It is believed that the unique mixotrophic nature of this organism, where it is able to feed on other phytoplankton, as well as photosynthesize by hosting a large endosymbiotic population of Pedinomonas noctilucae, gives it a greater competitive advantage over other phytoplankton. It is believed that this physiological plasticity of Noctiluca is responsible for the species shift from a diatom dominated ecosystem of the past in the Arabian sea, to the current widespread blooms of Noctiluca. Here we have attempted to examine how Noctiluca are able to sustain themselves over an extended period when nutrients from winter convective mixing are completely exhausted in the euphotic zone. In this study we test the hypotheses, that new inputs of nutrients from anthropogenic sources such as sewage and land-based nutrient runoff as well as from thick aeolian dust storms from the nearby regions such as Oman and Iran allow Noctiluca to survive and grow from winter into the spring inter-monsoon season. A recently cultured strain of Arabian Sea green Noctiluca grown in media amended with dust collected from Oman and Iran, different trace metals (Iron, Selenium and Cobalt) as well as nutrients (Nitrate, Ammonium and Urea) has been used to test our hypotheses. This is the first time such experiments have been conducted in the laboratory as previously no Noctiluca culture survived beyond 6 months. Growth of Noctiluca was measured in terms of cell count and the photosynthetic efficiency of cells using the fluorescence induction and relaxation technique which deduces the quantum efficiency of photochemistry in PSII. Moreover, we also measured the ammonia content in the cells which has long been suspected to provide buoyancy to cells and/or serve as a nitrogenous nutrient to the green endosymbionts that Noctiluca harbors.
Polybrominated Diphenyl Ethers in Dryer Lint: An Advanced Analysis Laboratory
ERIC Educational Resources Information Center
Thompson, Robert Q.
2008-01-01
An advanced analytical chemistry laboratory experiment is described that involves environmental analysis and gas chromatography-mass spectrometry. Students analyze lint from clothes dryers for traces of flame retardant chemicals, polybrominated diphenylethers (PBDEs), compounds receiving much attention recently. In a typical experiment, ng/g…
Experiment to Characterize Aircraft Volatile Aerosol and Trace-Species Emissions (EXCAVATE)
NASA Technical Reports Server (NTRS)
Anderson, B. E.; Branham, H.-S.; Hudgins, C. H.; Plant, J. V.; Ballenthin, J. O.; Miller, T. M.; Viggiano, A. A.; Blake, D. R.; Boudries, H.; Canagaratna, M.
2005-01-01
The Experiment to Characterize Aircraft Volatile and Trace Species Emissions (EXCAVATE) was conducted at Langley Research Center (LaRC) in January 2002 and focused upon assaying the production of aerosols and aerosol precursors by a modern commercial aircraft, the Langley B757, during ground-based operation. Remaining uncertainty in the postcombustion fate of jet fuel sulfur contaminants, the need for data to test new theories of particle formation and growth within engine exhaust plumes, and the need for observations to develop air quality models for predicting pollution levels in airport terminal areas were the primary factors motivating the experiment. NASA's Atmospheric Effects of Aviation Project (AEAP) and the Ultra Effect Engine Technology (UEET) Program sponsored the experiment which had the specific objectives of determining ion densities; the fraction of fuel S converted from S(IV) to S(VI); the concentration and speciation of volatile aerosols and black carbon; and gas-phase concentrations of long-chain hydrocarbon and PAH species, all as functions of engine power, fuel composition, and plume age.
Qiang, Hong; Lang, Dong-Li; Li, Yu-You
2012-01-01
The effect of trace metals on the mesophilic methane fermentation of high-solid food waste was investigated using both batch and continuous experiments. The continuous experiment was conducted by using a CSTR-type reactor with three run. During the first run, the HRT of the reactor was stepwise decreased from 100 days to 30 days. From operation day 50, the reactor efficiency deteriorated due to the lack of trace metals. The batch experiment showed that iron, cobalt, and nickel combinations had a significant effect on food waste. According to the results of the batch experiment, a combination of iron, cobalt, and nickel was added into the CSTR reactor by two different methods at run II, and III. Based on experimental results and theoretical calculations, the most suitable values of Fe/COD, Co/COD, and Ni/COD in the substrate were identified as 200, 6.0, and 5.7 mg/kg COD, respectively. Copyright © 2011 Elsevier Ltd. All rights reserved.
Morimoto, Y; Durante, W; Lancaster, D G; Klattenhoff, J; Tittel, F K
2001-01-01
Carbon monoxide (CO) has been implicated as a biological messenger molecule analogous to nitric oxide. A compact gas sensor based on a midinfrared laser absorption spectroscopy was developed for direct and real-time measurement of trace levels (in approximate pmol) of CO release by vascular cells. The midinfrared light is generated by difference frequency mixing of two nearinfrared lasers in a nonlinear optical crystal. A strong infrared absorption line of CO (4.61 microm) is chosen for convenient CO detection without interference from other gas species. The generation of CO from cultured vascular smooth muscle cells was detected every 20 s without any chemical modification to the CO. The sensitivity of the sensor reached 6.9 pmol CO. CO synthesis was measured from untreated control cells (0.25 nmol per 10(7) cells/h), sodium nitroprusside-treated cells (0.29 nmol per 10(7) cells/h), and hemin-treated cells (0.49 nmol per 10(7) cells/h). The sensor also detected decreases in CO production after the addition of the heme oxygenase (HO) inhibitor tin protoporphyrin-IX (from 0.49 to 0.02 nmol per 10(7) cells/h) and increases after the administration of the HO substrate hemin (from 0.27 to 0.64 nmol per 10(7) cells/h). These results demonstrate that midinfrared laser absorption spectroscopy is a useful technique for the noninvasive and real-time detection of trace levels of CO from biological tissues.
NASA Technical Reports Server (NTRS)
Morimoto, Y.; Durante, W.; Lancaster, D. G.; Klattenhoff, J.; Tittel, F. K.
2001-01-01
Carbon monoxide (CO) has been implicated as a biological messenger molecule analogous to nitric oxide. A compact gas sensor based on a midinfrared laser absorption spectroscopy was developed for direct and real-time measurement of trace levels (in approximate pmol) of CO release by vascular cells. The midinfrared light is generated by difference frequency mixing of two nearinfrared lasers in a nonlinear optical crystal. A strong infrared absorption line of CO (4.61 microm) is chosen for convenient CO detection without interference from other gas species. The generation of CO from cultured vascular smooth muscle cells was detected every 20 s without any chemical modification to the CO. The sensitivity of the sensor reached 6.9 pmol CO. CO synthesis was measured from untreated control cells (0.25 nmol per 10(7) cells/h), sodium nitroprusside-treated cells (0.29 nmol per 10(7) cells/h), and hemin-treated cells (0.49 nmol per 10(7) cells/h). The sensor also detected decreases in CO production after the addition of the heme oxygenase (HO) inhibitor tin protoporphyrin-IX (from 0.49 to 0.02 nmol per 10(7) cells/h) and increases after the administration of the HO substrate hemin (from 0.27 to 0.64 nmol per 10(7) cells/h). These results demonstrate that midinfrared laser absorption spectroscopy is a useful technique for the noninvasive and real-time detection of trace levels of CO from biological tissues.
Chow, Cynthia L.; Guo, Weixiang; Trivedi, Parul; Zhao, Xinyu; Gubbels, Samuel P.
2015-01-01
Hair cells in the adult mammalian cochlea cannot spontaneously regenerate after damage resulting in the permanency of hearing loss. Stem cells have been found to be present in the cochlea of young rodents; however, there has been little evidence for their existence into adulthood. We used nestin-CreERT2/tdTomato-reporter mice to trace the lineage of putative nestin-expressing cells and their progeny in the cochleae of adult mice. Nestin, an intermediate filament found in neural progenitor cells during early development and adulthood, is regarded as a multi-potent and neural stem cell marker. Other investigators have reported its presence in postnatal and young adult rodents; however, there are discrepancies amongst these reports. Using lineage tracing, we documented a robust population of tdTomato-expressing cells and evaluated these cells at a series of adult time points. Upon activation of the nestin promoter, tdTomato was observed just below and medial to the inner hair cell layer. All cells co-localized with the stem cell and cochlear-supporting-cell marker Sox2 as well as the supporting cell and Schwann cell marker Sox10; however, they did not co-localize with the Schwann cell marker Krox20, spiral ganglion marker NF200, or GFAP-expressing supporting cell marker. The cellular identity of this unique population of tdTomato-expressing cells in the adult cochlea of nestin-CreERT2/tdTomato mice remains unclear however these cells may represent a type of supporting cell on the neural aspect of the inner hair cell layer. PMID:25611038
Seress, L
1998-06-01
Jean Piaget's "stage theory" suggests that cognitive development proceeds in discrete steps, among which the first is the sensorimotor period that occupies the first two years. In recent years it became clear that an intact and mature hippocampus is necessary for memory formation both in experimental animals and in human. In the present experiments the perinatal morphological development of the human hippocampus was studied to describe structural changes that may correlate with the developmental changes of intellectual growth. Our results suggest that cell formation in the human hippocampus terminates several weeks before birth, but immature cells migrate to their final positions through the first six postnatal months. The newborn hippocampus contains all cell types and cell layers that are characteristic for the adult hippocampus. However, changes of the light microscopic features of the postsynaptic target neurons of hippocampal granule cells indicate that connections between granule cells and their target neurons are immature at birth and develop through an extended period of time that may last for three years. Since this neuronal connection is the first link in the chain of the main hippocampal synaptic circuitry, it may be suggested that human hippocampus is functionally impaired at birth. This period of light microscopic morphological maturation correlates well with the time period of Piaget's first stage of cognitive development. It can also be suggested that the prolonged postnatal development of some neuronal circuitries in the human hippocampus may be responsible for the psychological phenomenon of "infantile amnesia", that is the lack of memory traces from the early postnatal period.
Parabrachial complex links pain transmission to descending pain modulation.
Roeder, Zachary; Chen, QiLiang; Davis, Sophia; Carlson, Jonathan D; Tupone, Domenico; Heinricher, Mary M
2016-12-01
The rostral ventromedial medulla (RVM) has a well-documented role in pain modulation and exerts antinociceptive and pronociceptive influences mediated by 2 distinct classes of neurons, OFF-cells and ON-cells. OFF-cells are defined by a sudden pause in firing in response to nociceptive inputs, whereas ON-cells are characterized by a "burst" of activity. Although these reflex-related changes in ON- and OFF-cell firing are critical to their pain-modulating function, the pathways mediating these responses have not been identified. The present experiments were designed to test the hypothesis that nociceptive input to the RVM is relayed through the parabrachial complex (PB). In electrophysiological studies, ON- and OFF-cells were recorded in the RVM of lightly anesthetized male rats before and after an infusion of lidocaine or muscimol into PB. The ON-cell burst and OFF-cell pause evoked by noxious heat or mechanical probing were substantially attenuated by inactivation of the lateral, but not medial, parabrachial area. Retrograde tracing studies showed that neurons projecting to the RVM were scattered throughout PB. Few of these neurons expressed calcitonin gene-related peptide, suggesting that the RVM projection from PB is distinct from that to the amygdala. These data show that a substantial component of "bottom-up" nociceptive drive to RVM pain-modulating neurons is relayed through the PB. While the PB is well known as an important relay for ascending nociceptive information, its functional connection with the RVM allows the spinoparabrachial pathway to access descending control systems as part of a recurrent circuit.
DNA profiling of trace DNA recovered from bedding.
Petricevic, Susan F; Bright, Jo-Anne; Cockerton, Sarah L
2006-05-25
Trace DNA is often detected on handled items and worn clothing examined in forensic laboratories. In this study, the potential transfer of trace DNA to bedding by normal contact, when an individual sleeps in a bed, is examined. Volunteers slept one night on a new, lower bed sheet in their own bed and one night in a bed foreign to them. Samples from the sheets were collected and analysed by DNA profiling. The results indicate that the DNA profile of an individual can be obtained from bedding after one night of sleeping in a bed. The DNA profile of the owner of the bed could also be detected in the foreign bed experiments. Since mixed DNA profiles can be obtained from trace DNA on bedding, caution should be exercised when drawing conclusions from DNA profiling results obtained from such samples. This transfer may have important repercussions in sexual assault investigations.
Extinct Life on Mars: Looking for Traces of Viruses Instead of Bacteria
NASA Astrophysics Data System (ADS)
Ksanfomality, Leonid
The finds in the ALH 84001 meteorite were reported as a sign of traces and fossils of ancient Martian primitive life. Despite many experiments, the issue has not been settled. A study of geochemical data suggests that simple inorganic processes sometimes offer a more plausible explanation of the suspicious structures found in the ALH 84001 meteorite that had been discussed by many authors. A photo of certain micro structures got in the paper of N. Yushkin (Fig. 1) proves the point. (Figure: The inorganic structures in granites.) It is known that viruses possess an ability to survive under very severe external conditions. The Martian biogenic activity (if any) could leave its virus' sign. Thus it could make a sense to look for viruses or even DNA traces both in the body of the ALH 84001 meteorite and on Mars in future space missions instead of traces of the extinct bacteria.
Liu, Rongjun; Zhang, Liangliang; Chen, Yunyun; Huang, Zirong; Huang, Yong; Zhao, Shulin
2018-04-03
The superoxide anion (O 2 •- ) and hydroxyl radical ( • OH) are important reactive oxygen species (ROS) used as biomarkers in physiological and pathological processes. ROS generation is closely related to the development of a variety of inflammatory diseases. However, the changes of ROS are difficult to ascertain with in situ tracing of the inflammation process by real-time monitoring, owing to the short half-lives of ROS and high tissue autofluorescence in vivo. Here we developed a new near-infrared (NIR) ratiometric fluorescence imaging approach by using a Förster resonance energy transfer (FRET)-based ratiometric fluorescent nanoprobe for real-time monitoring of O 2 •- and • OH generation and also by using in situ tracing of the inflammation process in vivo. The proposed nanoprobe was composed of PEG functionalized GQDs as the energy donor connecting to hydroIR783, serving as both the O 2 •- / • OH recognizing ligand and the energy acceptor. The nanoprobe not only exhibited a fast response to O 2 •- and • OH but also presented good biocomapatibility as well as a high photostability and signal-to-noise ratio. We have demonstrated that the proposed NIR ratiometric fluorescent nanoprobe can monitor the changes of O 2 •- and • OH in living RAW 264.7 cells via a drug mediating inflammation model and further realized visual monitoring of the change of O 2 •- and • OH in mice for in situ tracing of the inflammation process. Our design may provide a new paradigm for long-term and real-time imaging applications for in vivo tracing of the pathological process related to the inflammatory diseases.
DOE Office of Scientific and Technical Information (OSTI.GOV)
Robertson, J.D.; Blanchard, L.J.; Srikantapura, S.
1996-12-31
The minor- and trace-element content of coal is of great interest because of the potentially hazardous impact on human health and the environment resulting from their release during coal combustion. Of the one billion tons of coal mined annually in the United States, 85-90% is consumed by coal-fired power plants. Potentially toxic elements present at concentrations as low as a few egg can be released in large quantities from combustion of this magnitude. Of special concern are those trace elements that occur naturally in coal which have been designated as potential hazardous air pollutants (HAPs) in the 1990 Amendments tomore » the Clean Air Act. The principle objective of this work was to investigate a combination of physical and chemical coal cleaning techniques to remove 90 percent of HAP trace elements at 90 percent combustibles recovery from Kentucky No. 9 coal. Samples of this coal were first subjected to physical separation by flotation in a Denver cell. The float fraction from the Denver cell was then used as feed material for hydrothermal leaching tests in which the efficacy of dilute alkali (NaOH) and acid (HNO{sub 3}) solutions at various temperatures and pressures was investigated. The combined column flotation and mild chemical cleaning strategy removed 60-80% of trace elements with greater than 85, recovery of combustibles from very finely ground (-325 mesh) coal. The elemental composition of the samples generated at each stage was determined using particle induced X-ray emission (PIXE) analysis. PIXE is a rapid, instrumental technique that, in principle, is capable of analyzing all elements from sodium through uranium with sensitivities as low as 1 {mu}g/g.« less
Pardo, Tania; Bes, Cleménce; Bernal, Maria Pilar; Clemente, Rafael
2016-11-01
Tailings are considered one of the most relevant sources of contamination associated with mining activities. Phytostabilization of mine spoils may need the application of the adequate combination of amendments to facilitate plant establishment and reduce their environmental impact. Two pot experiments were set up to assess the capability of 2 inorganic materials (calcium carbonate and a red mud derivate, ViroBind TM ), alone or in combination with organic amendments, for the stabilization of highly acidic trace element-contaminated mine tailings using Atriplex halimus. The effects of the treatments on tailings and porewater physico-chemical properties and trace-element accumulation by the plants, as well as the processes governing trace elements speciation and solubility in soil solution and their bioavailability were modeled. The application of the amendments increased tailings pH and decreased (>99%) trace elements solubility in porewater, but also changed the speciation of soluble Cd, Cu, and Pb. All the treatments made A. halimus growth in the tailings possible; organic amendments increased plant biomass and nutritional status, and reduced trace-element accumulation in the plants. Tailings amendments modified trace-element speciation in porewater (favoring the formation of chlorides and/or organo-metallic forms) and their solubility and plant uptake, which were found to be mainly governed by tailing/porewater pH, electrical conductivity, and organic carbon content, as well as soluble/available trace-element concentrations. Environ Toxicol Chem 2016;35:2874-2884. © 2016 SETAC. © 2016 SETAC.
Measurements of trace contaminants in closed-type plant cultivation chambers
NASA Astrophysics Data System (ADS)
Tani, A.; Kiyota, M.; Aiga, I.; Nitta, K.; Tako, Y.; Ashida, A.; Otsubo, K.; Saito, T.
Trace contaminants generated in closed facilities can cause abnormal plant growth. We present measurement data of trace contaminants released from soils, plants, and construction materials. We mainly used two closed chambers, a Closed-type Plant and Mushroom Cultivation Chamber (PMCC) and Closed-type Plant Cultivation Equipment (CPCE). Although trace gas budgets from soils obtained in this experiment are only one example, the results indicate that the budgets of trace gases, as well as CO_2 and O_2, change greatly with the degree of soil maturation and are dependent on the kind of substances in the soil. Both in the PMCC and in the CPCE, trace gases such as dioctyl phthalate (DOP), dibutyl phthalate (DBP), toluene and xylene were detected. These gases seemed to be released from various materials used in the construction of these chambers. The degree of increase in these trace gas levels was dependent on the relationship between chamber capacity and plant quantity. Results of trace gas measurement in the PMCC, in which lettuce and shiitake mushroom were cultivated, showed that ethylene was released both from lettuce and from the mushroom culture bed. The release rates were about 90 ng bed^-1 h^-1 for the shiitake mushroom culture bed (volume is 1700 cm^3) and 4.1 ~ 17.3 ng dm^-2h^-1 (leaf area basis) for lettuce. Higher ethylene release rates per plant and per unit leaf area were observed in mature plants than in young plants.
Verification technology of remote sensing camera satellite imaging simulation based on ray tracing
NASA Astrophysics Data System (ADS)
Gu, Qiongqiong; Chen, Xiaomei; Yang, Deyun
2017-08-01
Remote sensing satellite camera imaging simulation technology is broadly used to evaluate the satellite imaging quality and to test the data application system. But the simulation precision is hard to examine. In this paper, we propose an experimental simulation verification method, which is based on the test parameter variation comparison. According to the simulation model based on ray-tracing, the experiment is to verify the model precision by changing the types of devices, which are corresponding the parameters of the model. The experimental results show that the similarity between the imaging model based on ray tracing and the experimental image is 91.4%, which can simulate the remote sensing satellite imaging system very well.
Graded levels of Pax2a and Pax8 regulate cell differentiation during sensory placode formation.
McCarroll, Matthew N; Lewis, Zachary R; Culbertson, Maya Deza; Martin, Benjamin L; Kimelman, David; Nechiporuk, Alex V
2012-08-01
Pax gene haploinsufficiency causes a variety of congenital defects. Renal-coloboma syndrome, resulting from mutations in Pax2, is characterized by kidney hypoplasia, optic nerve malformation, and hearing loss. Although this underscores the importance of Pax gene dosage in normal development, how differential levels of these transcriptional regulators affect cell differentiation and tissue morphogenesis is still poorly understood. We show that differential levels of zebrafish Pax2a and Pax8 modulate commitment and behavior in cells that eventually contribute to the otic vesicle and epibranchial placodes. Initially, a subset of epibranchial placode precursors lie lateral to otic precursors within a single Pax2a/8-positive domain; these cells subsequently move to segregate into distinct placodes. Using lineage-tracing and ablation analyses, we show that cells in the Pax2a/8+ domain become biased towards certain fates at the beginning of somitogenesis. Experiments involving either Pax2a overexpression or partial, combinatorial Pax2a and Pax8 loss of function reveal that high levels of Pax favor otic differentiation whereas low levels increase cell numbers in epibranchial ganglia. In addition, the Fgf and Wnt signaling pathways control Pax2a expression: Fgf is necessary to induce Pax2a, whereas Wnt instructs the high levels of Pax2a that favor otic differentiation. Our studies reveal the importance of Pax levels during sensory placode formation and provide a mechanism by which these levels are controlled.
Graded levels of Pax2a and Pax8 regulate cell differentiation during sensory placode formation
McCarroll, Matthew N.; Lewis, Zachary R.; Culbertson, Maya Deza; Martin, Benjamin L.; Kimelman, David; Nechiporuk, Alex V.
2012-01-01
Pax gene haploinsufficiency causes a variety of congenital defects. Renal-coloboma syndrome, resulting from mutations in Pax2, is characterized by kidney hypoplasia, optic nerve malformation, and hearing loss. Although this underscores the importance of Pax gene dosage in normal development, how differential levels of these transcriptional regulators affect cell differentiation and tissue morphogenesis is still poorly understood. We show that differential levels of zebrafish Pax2a and Pax8 modulate commitment and behavior in cells that eventually contribute to the otic vesicle and epibranchial placodes. Initially, a subset of epibranchial placode precursors lie lateral to otic precursors within a single Pax2a/8-positive domain; these cells subsequently move to segregate into distinct placodes. Using lineage-tracing and ablation analyses, we show that cells in the Pax2a/8+ domain become biased towards certain fates at the beginning of somitogenesis. Experiments involving either Pax2a overexpression or partial, combinatorial Pax2a and Pax8 loss of function reveal that high levels of Pax favor otic differentiation whereas low levels increase cell numbers in epibranchial ganglia. In addition, the Fgf and Wnt signaling pathways control Pax2a expression: Fgf is necessary to induce Pax2a, whereas Wnt instructs the high levels of Pax2a that favor otic differentiation. Our studies reveal the importance of Pax levels during sensory placode formation and provide a mechanism by which these levels are controlled. PMID:22745314
Effect of silicon on trace element partitioning in iron-bearing metallic melts
NASA Astrophysics Data System (ADS)
Chabot, Nancy L.; Safko, Trevor M.; McDonough, William F.
2010-08-01
Despite the fact that Si is considered a potentially important metalloid in planetary systems, little is known about the effect of Si in metallic melts on trace element partitioning behavior. Previous studies have established the effects of S, C, and P, nonmetals, through solid metal/liquid metal experiments in the corresponding Fe binary systems, but the Fe-Si system is not appropriate for similar experiments because of the high solubility of Si in solid metal. In this work, we present the results from 0.1MPa experiments with two coexisting immiscible metallic liquids in the Fe-S-Si system. By leveraging the extensive available knowledge about the effect of S on trace element partitioning behavior, we explore the effect of Si. Results for 22 trace elements are presented. Strong Si avoidance behavior is demonstrated by As, Au, Ga, Ge, Sb, Sn, and Zn. Iridium, Os, Pt, Re, Ru, and W exhibit weak Si avoidance tendencies. Silicon appears to have no significant effect on the partitioning behaviors of Ag, Co, Cu, Cr, Ni, Pd, and V, all of which had similar partition coefficients over a wide range of Si liquid concentrations from Si-free to 13 wt%. The only elements in our experiments to show evidence of a potentially weak attraction to Si were Mo and Rh. Applications of the newly determined effects of Si to problems in planetary science indicate that (1) The elements Ni, Co, Mo, and W, which are commonly used in planetary differentiation models, are minimally affected by the presence of Si in the metal, especially in comparison to other effects such as from oxygen fugacity. 2) Reduced enstatite-rich meteorites may record a chemical signature due to Si in the metallic melts during partial melting, and if so, elements identified by this study as having strong Si avoidance may offer unique insight into unraveling the history of these meteorites.
ACS experiment for atmospheric studies on "ExoMars-2016" Orbiter
NASA Astrophysics Data System (ADS)
Korablev, O. I.; Montmessin, F.; Fedorova, A. A.; Ignatiev, N. I.; Shakun, A. V.; Trokhimovskiy, A. V.; Grigoriev, A. V.; Anufreichik, K. A.; Kozlova, T. O.
2015-12-01
ACS is a set of spectrometers for atmospheric studies (Atmospheric Chemistry Suite). It is one of the Russian instruments for the Trace Gas Orbiter (TGO) of the Russian-European "ExoMars" program. The purpose of the experiment is to study the Martian atmosphere by means of two observations regimes: sensitive trace gases measurements in solar occultations and by monitoring the atmospheric state during nadir observations. The experiment will allow us to approach global problems of Mars research such as current volcanism, and the modern climate status and its evolution. Also, the experiment is intended to solve the mystery of methane presence in the Martian atmosphere. Spectrometers of the ACS set cover the spectral range from the near IR-range (0.7 μm) to the thermal IR-range (17 μm) with spectral resolution λ/Δλ reaching 50000. The ACS instrument consists of three independent IR spectrometers and an electronics module, all integrated in a single unit with common mechanical, electrical and thermal interfaces. The article gives an overview of scientific tasks and presents the concept of the experiment.
Trace element distribution in the rat cerebellum
NASA Astrophysics Data System (ADS)
Kwiatek, W. M.; Long, G. J.; Pounds, J. G.; Reuhl, K. R.; Hanson, A. L.; Jones, K. W.
1990-04-01
Spatial distributions and concentrations of trace elements (TE) in the brain are important because TE perform catalytic and structural functions in enzymes which regulate brain function and development. We have investigated the distributions of TE in rat cerebellum. Structures were sectioned and analyzed by the Synchrotron Radiation Induced X-ray Emission (SRIXE) method using the NSLS X-26 white-light microprobe facility. Advantages important for TE analysis of biological specimens with X-ray microscopy include short time of measurement, high brightness and flux, good spatial resolution, multielemental detection, good sensitivity, and nondestructive irradiation. Trace elements were measured in thin rat brain sections of 20 μm thickness. The analyses were performed on sample volumes as small as 0.2 nl with Minimum Detectable Limits (MDL) of 50 ppb wet weight for Fe, 100 ppb wet weight for Cu, and Zn, and 1 ppm wet weight for Pb. The distribution of TE in the molecular cell layer, granule cell layer and fiber tract of rat cerebella was investigated. Both point analyses and two-dimensional semiquantitative mapping of the TE distribution in a section were used. All analyzed elements were observed in each structure of the cerebellum except mercury which was not observed in granule cell layer or fiber tract. This approach permits an exacting correlation of the TE distribution in complex structure with the diet, toxic elements, and functional status of the animal.
Application of a broadly tunable SG-DBR QCL for multi-species trace gas spectroscopy.
Diba, Abdou S; Xie, Feng; Gross, Barry; Hughes, Lawrence C; Zah, Chung-en; Moshary, Fred
2015-10-19
Feasibility of using a mid-Infrared tunable sampled-grating distributed Bragg reflectors quantum cascade laser for high resolution multicomponent trace gas spectroscopy is demonstrated. By controlling the driving currents to the front and back sections of the laser, we were able to tune a pulsed 4.55 µm laser over a frequency range a of 30 cm(-1) with high resolution, accuracy and repeatability. The laser was applied to absorption spectroscopy of ambient and reduced pressure (150 Torr) air in a 205 meters multi-pass Herriott cell, and by using standard LSQ fitting to a spectral database of these trace gases (HITRAN), the concentrations of nitrous oxide, carbon monoxide, and water vapor were retrieved.
A Multi-Scale Study on the Role of Trace Metals on Physiological and Pathological Mineralization
NASA Astrophysics Data System (ADS)
Rammelkamp, Derek
The work in this thesis provides mulit-scale contributions towards understanding the effects of trace metals on the pathological mineralization process relating to both the development of healthy bone tissue, the diseased state of osteoporosis, and microcalcifications which develop in breast cancers. A protein level study was performed on ECM protein fibronectin, which plays a role in cell adhesion. The protein studies showed zinc interactions with fibronectin and its fragment, anastellin, to influence protein structure. Zinc is also shown to decrease cell migration in vitro, which may be influenced by changes in fibronectin ECM structure. The effects of osteoporosis on micronutrient composition in vivo were examined using the technique of x-ray fluorescence (XRF) in an ovariectomized rat model. Compared to healthy bone, subtle difference are observed in zinc and iron in osteoporotic rat bones, showing micronutrients may play an important role in healthy bone regulation. Effects of micronutrient zinc was used to inhibit microcalcification formation in breast cancers. Microcalcifications have been linked malignancy of breast cancers, but the process of microcalcification formation has yet to be well understood. In this work, exogenous zinc is used to inhibit microcalcification formation, and metastatic potential in both a 2D and 3D spheroid environment. A novel in vitro self-assembled three dimensional multi-cellular tumor spheroid (MCTS) model for the study of breast cancer microcalcifications was developed for this experiment. A MCTS model for studying breast cancer microcalcifications has potential to be used in drug discovery, or for basic research applications studying mechanisms of microcalcification formation, which are still not fully understood. Taken together this study uses a multi-scale approach to gain a better understanding of micronutrients involved in pathological mineralization.
Novel Combinatory Approaches to Repair Visual System after Optic Nerve Damage
2014-09-01
Intravitreal Viral Treatment Outcome Measures PTEN/SOCS3f/f AAV-GFP (Control) RGC staining and axon tracing PTEN/SOCS3f/f AAV-Cre “ “ CHOP-/- AAV-XBP1...survival and axon regeneration using immunohistochemistry and axon tracing methods used in our previous studies. At 4 and 8 weeks after injury, retinas...the cell body. After peripheral nerve injury, phosphorylated (i.e., active) STAT3 is detected in the injured axons and later in the soma (Lee, Neitzel
Regular Biology Students Learn Like AP Students with SUN
ERIC Educational Resources Information Center
Batiza, Ann; Luo, Wen; Zhang, Bo; Gruhl, Mary; Nelson, David; Hoelzer, Mark; Ning, Ling; Roberts, Marisa; Knopp, Jonathan; Harrington, Tom; LaFlamme, Donna; Haasch, Mary Anne; Vogt, Gina; Goodsell, David; Marcey, David
2016-01-01
The SUN approach to biological energy transfer education is fundamentally different from past practices that trace chemical and energy inputs and outputs. The SUN approach uses a hydrogen fuel cell to convince learners that electrons can move from one substance to another based on differential attraction. With a hydrogen fuel cell, learners can…
Ye, Da-Wei; Liu, Cheng; Tian, Xue-Bi; Xiang, Hong-Bing
2014-01-01
To determine the spinal innervation and neuronal connections is important for studying gastric carbohydrate metabolism and motor responses. Neurons involved in the efferent control of the stomach were identified following visualization of pseudorabies virus (PRV)-614 retrograde tracing. PRV-614 was injected into the ventral stomach wall in 13 adult C57BL/6J strain male mice. On the fifth day postinjection, animals were humanely sacrificed, and spinal cords were removed and sectioned, and processed for PRV visualization. The virus injected into the ventral stomach wall was specifically transported to the thoracic spinal cord. At 5 d after injection of the PRV-614, stomach enlargement and tissue edema were found, and PRV-614 positive cells were found in the intermediolateral cell column, the intercalates nucleus or the central autonomic nucleus of spinal cord segments T3 to L1, and major PRV-614 labeled cells were focused in the T6-10 segment. Our results revealed neuroanatomical circuits between stomach and the spinal intermediolateral cell column neurons.
ERIC Educational Resources Information Center
Hodgson, Ted; Andersen, Lyle; Robison-Cox, Jim; Jones, Clain
2004-01-01
Water quality experiments, especially the use of macroinvertebrates as indicators of water quality, offer an ideal context for connecting statistics and science. In the STAR program for secondary students and teachers, water quality experiments were also used as a context for teaching statistics. In this article, we trace one activity that uses…
High Resolution Numerical Simulations and Diagnostic Studies Atmospheric Transport During TRACE-A
NASA Technical Reports Server (NTRS)
Krishnamurti, T. N.; Fuelberg, Henry
1995-01-01
The enclosed publications constitute our final report. These are publications completed in referred journals. The completed work includes: Meteorological conditions associated with vertical distributions of aerosols off the west coast of Africa. TRACE-A trajectory intercomparison 2. Isentropic and kinematic methods. Chemical characteristics of tropospheric air over the tropical South Atlantic Ocean: Relationship to trajectory history. Passive Tracer Transport Relevant to the TRACE-A Experiment. The meteorological environment of the tropospheric ozone maximum over the tropical south Atlantic Ocean. : Influence of a middle-latitude cyclone on tropospheric ozone distributions during a period of TRACE-A. All of this work provided the meteorological background for the Transport of Atmospheric Chemistry near the Equator (TRACE-A) project sponsored by Dr,, Joe McNeal of NASA. Our principal findings are that bio-mass burning sources from Africa and South America did contribute to the accumulation of tropospheric ozone over the tropical Atlantic Ocean. Other findings relate to circulations, advections of Ozone and other plausible sources for the TOMS based ozone maximum over the Atlantic Ocean.
Khan, Naeem; Choi, Ji Yeon; Nho, Eun Yeong; Jamila, Nargis; Habte, Girum; Hong, Joon Ho; Hwang, In Min; Kim, Kyong Su
2014-09-01
This study aimed at analyzing the concentrations of 23 minor and trace elements in aromatic spices by inductively coupled plasma-mass spectrometry (ICP-MS), after wet digestion by microwave system. The analytical method was validated by linearity, detection limits, precision, accuracy and recovery experiments, obtaining satisfactory values in all cases. Results indicated the presence of variable amounts of both minor and trace elements in the selected aromatic spices. Manganese was high in cinnamon (879.8 μg/g) followed by cardamom (758.1 μg/g) and clove (649.9 μg/g), strontium and zinc were high in ajwain (489.9 μg/g and 84.95 μg/g, respectively), while copper was high in mango powder (77.68 μg/g). On the whole some of the minor and essential trace elements were found to have good nutritional contribution in accordance to RDA. The levels of toxic trace elements, including As, Cd, and Pb were very low and did not found to pose any threat to consumers. Copyright © 2014 Elsevier Ltd. All rights reserved.
NASA Technical Reports Server (NTRS)
Gerrard, Andrew J.; Kane, Timothy J.; Eckermann, Stephen D.; Thayer, Jeffrey P.
2004-01-01
We conducted gravity wave ray-tracing experiments within an atmospheric region centered near the ARCLITE lidar system at Sondrestrom, Greenland (67N, 310 deg E), in efforts to understand lidar observations of both upper stratospheric gravity wave activity and mesospheric clouds during August 1996 and the summer of 2001. The ray model was used to trace gravity waves through realistic three-dimensional daily-varying background atmospheres in the region, based on forecasts and analyses in the troposphere and stratosphere and climatologies higher up. Reverse ray tracing based on upper stratospheric lidar observations at Sondrestrom was also used to try to objectively identify wave source regions in the troposphere. A source spectrum specified by reverse ray tracing experiments in early August 1996 (when atmospheric flow patterns produced enhanced transmission of waves into the upper stratosphere) yielded model results throughout the remainder of August 1996 that agreed best with the lidar observations. The model also simulated increased vertical group propagation of waves between 40 km and 80 km due to intensifying mean easterlies, which allowed many of the gravity waves observed at 40 km over Sondrestrom to propagate quasi-vertically from 40-80 km and then interact with any mesospheric clouds at 80 km near Sondrestrom, supporting earlier experimentally-inferred correlations between upper stratospheric gravity wave activity and mesospheric cloud backscatter from Sondrestrom lidar observations. A pilot experiment of real-time runs with the model in 2001 using weather forecast data as a low-level background produced less agreement with lidar observations. We believe this is due to limitations in our specified tropospheric source spectrum, the use of climatological winds and temperatures in the upper stratosphere and mesosphere, and missing lidar data from important time periods.
Forensic Excavation of Rock Masses: A Technique to Investigate Discontinuity Persistence
NASA Astrophysics Data System (ADS)
Shang, J.; Hencher, S. R.; West, L. J.; Handley, K.
2017-11-01
True persistence of rock discontinuities (areas with insignificant tensile strength) is an important factor controlling the engineering behaviour of fractured rock masses, but is extremely difficult to quantify using current geological survey methodologies, even where there is good rock exposure. Trace length as measured in the field or using remote measurement devices is actually only broadly indicative of persistence for rock engineering practice and numerical modelling. Visible traces of discontinuities are treated as if they were open fractures within rock mass classifications, despite many such traces being non-persistent and actually retaining considerable strength. The common assumption of 100% persistence, based on trace length, is generally extremely conservative in terms of strength and stiffness, but not always so and may lead to a wrong prediction of failure mechanism or of excavatability. Assuming full persistence would give hopelessly incorrect predictions of hydraulic conductivity. A new technique termed forensic excavation of rock masses is introduced, as a procedure for directly investigating discontinuity persistence. This technique involves non-explosive excavation of rock masses by injecting an expansive chemical splitter along incipient discontinuities. On expansion, the splitter causes the incipient traces to open as true joints. Experiments are described in which near-planar rock discontinuities, through siltstone and sandstone, were opened up by injecting the splitter into holes drilled along the lines of visible traces of the discontinuities in the laboratory and in the field. Once exposed the surfaces were examined to investigate the pre-existing persistence characteristics of the incipient discontinuities. One conclusion from this study is that visible trace length of a discontinuity can be a poor indicator of true persistence (defined for a fracture area with negligible tensile strength). An observation from this series of experiments was that freshly failed surfaces through pre-existing rock bridges were relatively rough compared to sections of pre-existing weaker areas of geologically developed (though still incipient) discontinuities. Fractographic features such as hackle and rib marks were typical of the freshly broken rock bridges, whereas opened-up areas of incipient discontinuity were smoother. Schmidt hammer rebound values were generally higher for the rock bridge areas, probably reflecting their lower degree of chemical and physical weathering.
Tactual discrimination of softness.
Srinivasan, M A; LaMotte, R H
1995-01-01
1. We investigated the ability of humans to tactually discriminate the softness of objects, using novel elastic objects with deformable and rigid surfaces. For objects with deformable surfaces, we cast transparent rubber specimens with variable compliances. For objects with rigid surfaces ("spring cells") we fabricated telescoping hollow cylinders with the inner cylinder supported by several springs. To measure the human discriminability and to isolate the associated information-processing mechanisms, we performed psychophysical experiments under three conditions: 1) active touch with the normal finger, where both tactile and kinesthetic information was available to the subject: 2) active touch with local cutaneous anesthesia, so that only kinesthetic information was available; and 3) passive touch, where a computer-controlled mechanical stimulator brought down the compliant specimens onto the passive fingerpad of the subject, who therefore had only tactile information. 2. We first characterized the mechanical behavior of the human fingerpad and the test objects by determining the relationship between the depth and force of indentation during constant-velocity indentations by a rigid probe. The fingerpad exhibited a pronounced nonlinear behavior in the indentation depth versus force trace such that compliance, as indicated by the local slope of the trace, decreased with increases in indentation depth. The traces for all the rubber specimens were approximately linear, indicating a constant but distinct value of compliance for each specimen. The fingerpad was more compliant than each of the rubber specimens. 3. All the human subjects showed excellent softness discriminability in ranking the rubber specimens by active touch, and the subjective perception of softness correlated one-to-one with the objectively measured compliance. The ability of subjects to discriminate the compliance of spring cells was consistently poorer compared with that of the rubber specimens. 4. For pairwise discrimination of a selected set of rubber specimens, kinesthetic information alone was insufficient. However, tactile information alone was sufficient, even when the velocities and forces of specimen application were randomized. In contrast, for discriminating pairs of spring cells, tactile information alone was insufficient, and both tactile and kinesthetic information were found to be necessary. 5. The differences in the sufficiency of tactile information for the discrimination of the two types of objects can be explained by the mechanics of contact of the fingerpad and its effect on tactile information. For objects with deformable surfaces, the spatial pressure distribution within the contact region depends on both the force applied and the specimen compliance.(ABSTRACT TRUNCATED AT 250 WORDS)
ERIC Educational Resources Information Center
Beaulieu, Lionel J.; Barfield, Melissa
This study examines the link between human capital endowments of Southern workers and their labor force experiences over time. Using a national longitudinal survey, the experiences of 4,566 individuals who left high school in 1982 were traced through 1992. Findings show similar patterns of educational attainment between women and men, but African…
Milgram's Obedience to Authority experiments: origins and early evolution.
Russell, Nestar John Charles
2011-03-01
Stanley Milgram's Obedience to Authority experiments remain one of the most inspired contributions in the field of social psychology. Although Milgram undertook more than 20 experimental variations, his most (in)famous result was the first official trial run - the remote condition and its 65% completion rate. Drawing on many unpublished documents from Milgram's personal archive at Yale University, this article traces the historical origins and early evolution of the obedience experiments. Part 1 presents the previous experiences that led to Milgram's conception of his rudimentary research idea and then details the role of his intuition in its refinement. Part 2 traces the conversion of Milgram's evolving idea into a reality, paying particular attention to his application of the exploratory method of discovery during several pilot studies. Both parts illuminate Milgram's ad hoc introduction of various manipulative techniques and subtle tension-resolving refinements. The procedural adjustments continued until Milgram was confident that the first official experiment would produce a high completion rate, a result contrary to expectations of people's behaviour. Showing how Milgram conceived of, then arrived at, this first official result is important because the insights gained may help others to determine theoretically why so many participants completed this experiment. ©2010 The British Psychological Society.
NASA Technical Reports Server (NTRS)
Gregory, Gerald L.; Scott, A. Donald, Jr.
1995-01-01
This compendium describes aircraft data that are available from NASA's Transport and Atmospheric Chemistry near the Equator - Atlantic (TRACE-A) conducted in September/October 1992. The broad objectives of TRACE-A were to study chemical processes and long-range transport associated with South American and African continental outflow during periods of widespread vegetation burning, and to understand the ozone enhancements observed from satellite data measured over the southern tropical Atlantic Ocean during the September/October time period. Flight experiments were conducted from Brazil, South Africa, Namibia, and the Ascension Island. This document provides a representation of aircraft data that are available from NASA Langley's Distributed Active Archive Center (DAAC). The data format of time series and altitude profile plots is not intended to support original analyses, but to assist the reader in identifying data that are of interest. This compendium is for only the NASA aircraft data. The DAAC data base includes numerous supporting data-meteorological products, results from surface studies, satellite observations, and data from sonde releases.
Measurement of OH, H2SO4, MSA, and HNO3 Aboard the P-3B Aircraft
NASA Technical Reports Server (NTRS)
Eisele, F. L.
2003-01-01
This paper addresses the measurement of OH, H2SO4, MSA, and HNO3 aboard the P-3B aircraft under the following headings: 1) Performance Report; 2) Highlights of OH, H2SO4, and MSA Measurements Made Aboard the NASA P-3B During TRACE-P; 3) Development and characteristics of an airborne-based instrument used to measure nitric acid during the NASA TRACE-P field experiment.
NASA Technical Reports Server (NTRS)
Farmer, Crofton B.; Raper, Odell F.
1987-01-01
The ATMOS (Atmospheric Trace Molecule Spectroscopy) experiment has the broad purpose of investigating the physical structure, chemistry, and dynamics of the upper atmosphere through the study of the distributions of the neutral minor and trace constituents and their seasonal and long-term variations. The technique used is high-resolution infrared absorption spectroscopy using the Sun as the radiation source, observing the changes in the transmission of the atmosphere as the line-of-sight from the Sun to the spacecraft penetrates the atmosphere close to the Earth's limb at sunrise and sunset. During these periods, interferograms are generated at the rate of one each second which yield, when transformed, high resolution spectra covering the 2.2 to 16 micron region of the infrared. Twenty such occultations were recorded during the Spacelab 3 flight, which have produced concentration profiles for a large number of minor and trace upper atmospheric species in both the Northern and Southern Hemispheres. Several of these species have not previously been observed in spectroscopic data. The data reduction and analysis procedures used following the flight are discussed; a number of examples of the spectra obtained are shown, and a bar graph of the species detected thus far in the analysis is given which shows the altitude ranges for which concentration profiles were retrieved.
Immune cells tracing using quantum dots
NASA Astrophysics Data System (ADS)
Hoshino, Akiyoshi; Fujioka, Kouki; Kawamura, Yuki I.; Toyama-Sorimachi, Noriko; Yasuhara, Masato; Dohi, Taeko; Yamamoto, Kenji
2006-02-01
Fluorescent nanoparticles, such as nanocrystal quantum dots (QDs), have potential to be applied to molecular biology and bioimaging, since some nanocrystals emit higher and longer lasting fluorescence than conventional organic probes do. Here we report an example of labeling immune cells by QDs. We collected splenic CD4 + T-lymphocyte and peritoneal macrophages from mice. Then cells were labeled with QDs. QDs are incorporated into the T-lymphocyte and macrophages immediately after addition and located in the cytoplasm via endocytosis pathway. The fluorescence of QDs held in the endosomes was easily detected for more than a week. In addition, T-lymphocytes labeled with QDs were stable and cell proliferation or cytokine production including IL-2 and IFN-γ was not affected. When QD-labeled T-lymphocytes were adoptively transferred intravenously to mice, they remained in the peripheral blood and spleen up to a week. Using QD-labeled peritoneal macrophages, we studied cell traffic during inflammation on viscera in peritoneum cavity. QD-labeled macrophages were transplanted into the peritoneum of the mouse, and colitis was induced by intracolonic injection of a hapten, trinitrobenzensulfonic acid. With the aid of stong signals of QDs, we found that macrophage accumuled on the inflammation site of the colon. These results suggested that fluorescent probes of QDs might be useful as bioimaging tools for tracing target cells in vivo.
Fu, Dan; Yu, Yong; Folick, Andrew; Currie, Erin; Farese, Robert V; Tsai, Tsung-Huang; Xie, Xiaoliang Sunney; Wang, Meng C
2014-06-18
Metabolic fingerprinting provides valuable information on the physiopathological states of cells and tissues. Traditional imaging mass spectrometry and magnetic resonance imaging are unable to probe the spatial-temporal dynamics of metabolites at the subcellular level due to either lack of spatial resolution or inability to perform live cell imaging. Here we report a complementary metabolic imaging technique that is based on hyperspectral stimulated Raman scattering (hsSRS). We demonstrated the use of hsSRS imaging in quantifying two major neutral lipids: cholesteryl ester and triacylglycerol in cells and tissues. Our imaging results revealed previously unknown changes of lipid composition associated with obesity and steatohepatitis. We further used stable-isotope labeling to trace the metabolic dynamics of fatty acids in live cells and live Caenorhabditis elegans with hsSRS imaging. We found that unsaturated fatty acid has preferential uptake into lipid storage while saturated fatty acid exhibits toxicity in hepatic cells. Simultaneous metabolic fingerprinting of deuterium-labeled saturated and unsaturated fatty acids in living C. elegans revealed that there is a lack of interaction between the two, unlike previously hypothesized. Our findings provide new approaches for metabolic tracing of neutral lipids and their precursors in living cells and organisms, and could potentially serve as a general approach for metabolic fingerprinting of other metabolites.
Composition of commercial media used for human embryo culture.
Morbeck, Dean E; Krisher, Rebecca L; Herrick, Jason R; Baumann, Nikola A; Matern, Dietrich; Moyer, Thomas
2014-09-01
To determine the composition of commercially available culture media and test whether differences in composition are biologically relevant in a murine model. Experimental laboratory study. University-based laboratory. Cryopreserved hybrid mouse one-cell embryos were used in experiments. Amino acid, organic acid, ions, and metal content were determined for two different lots of media from Cook, In Vitro Care, Origio, Sage, Vitrolife, Irvine CSC, and Global. To determine whether differences in the composition of these media are biologically relevant, mouse one-cell embryos were thawed and cultured for 120 hours in each culture media at 5% and 20% oxygen in the presence or absence of protein in an EmbryoScope time-lapse incubator. The compositions of seven culture media were analyzed for concentrations of 39 individual amino acids, organic acids, ions, and elements. Blastocyst rates and cell cycle timings were calculated at 96 hours of culture, and the experiments were repeated in triplicate. Of the 39 analytes, concentrations of glucose, lactate, pyruvate, amino acids, phosphate, calcium, and magnesium were present in variable concentrations, likely reflecting differences in the interpretation of animal studies. Essential trace elements, such as copper and zinc, were not detected. Mouse embryos failed to develop in one culture medium and were differentially affected by oxygen in two other media. Culture media composition varies widely, with differences in pyruvate, lactate, and amino acids especially notable. Blastocyst development was culture media dependent and showed an interaction with oxygen concentration and presence of protein. Copyright © 2014 American Society for Reproductive Medicine. Published by Elsevier Inc. All rights reserved.
Neurophysiological bases of exponential sensory decay and top-down memory retrieval: a model.
Zylberberg, Ariel; Dehaene, Stanislas; Mindlin, Gabriel B; Sigman, Mariano
2009-01-01
Behavioral observations suggest that multiple sensory elements can be maintained for a short time, forming a perceptual buffer which fades after a few hundred milliseconds. Only a subset of this perceptual buffer can be accessed under top-down control and broadcasted to working memory and consciousness. In turn, single-cell studies in awake-behaving monkeys have identified two distinct waves of response to a sensory stimulus: a first transient response largely determined by stimulus properties and a second wave dependent on behavioral relevance, context and learning. Here we propose a simple biophysical scheme which bridges these observations and establishes concrete predictions for neurophsyiological experiments in which the temporal interval between stimulus presentation and top-down allocation is controlled experimentally. Inspired in single-cell observations, the model involves a first transient response and a second stage of amplification and retrieval, which are implemented biophysically by distinct operational modes of the same circuit, regulated by external currents. We explicitly investigated the neuronal dynamics, the memory trace of a presented stimulus and the probability of correct retrieval, when these two stages were bracketed by a temporal gap. The model predicts correctly the dependence of performance with response times in interference experiments suggesting that sensory buffering does not require a specific dedicated mechanism and establishing a direct link between biophysical manipulations and behavioral observations leading to concrete predictions.
Haldipur, Parthiv; Dang, Derek; Aldinger, Kimberly A; Janson, Olivia K; Guimiot, Fabien; Adle-Biasette, Homa; Dobyns, William B; Siebert, Joseph R; Russo, Rosa; Millen, Kathleen J
2017-01-01
FOXC1 loss contributes to Dandy-Walker malformation (DWM), a common human cerebellar malformation. Previously, we found that complete Foxc1 loss leads to aberrations in proliferation, neuronal differentiation and migration in the embryonic mouse cerebellum (Haldipur et al., 2014). We now demonstrate that hypomorphic Foxc1 mutant mice have granule and Purkinje cell abnormalities causing subsequent disruptions in postnatal cerebellar foliation and lamination. Particularly striking is the presence of a partially formed posterior lobule which echoes the posterior vermis DW 'tail sign' observed in human imaging studies. Lineage tracing experiments in Foxc1 mutant mouse cerebella indicate that aberrant migration of granule cell progenitors destined to form the posterior-most lobule causes this unique phenotype. Analyses of rare human del chr 6p25 fetal cerebella demonstrate extensive phenotypic overlap with our Foxc1 mutant mouse models, validating our DWM models and demonstrating that many key mechanisms controlling cerebellar development are likely conserved between mouse and human. DOI: http://dx.doi.org/10.7554/eLife.20898.001 PMID:28092268
Xu, Jin; Cui, Jiaxi; Del Campo, Aranzazu; Shin, Chong Hyun
2016-01-01
The liver and pancreas originate from overlapping embryonic regions, and single-cell lineage tracing in zebrafish has shown that Bone morphogenetic protein 2b (Bmp2b) signaling is essential for determining the fate of bipotential hepatopancreatic progenitors towards the liver or pancreas. Despite its pivotal role, the gene regulatory networks functioning downstream of Bmp2b signaling in this process are poorly understood. We have identified four and a half LIM domains 1b (fhl1b), which is primarily expressed in the prospective liver anlage, as a novel target of Bmp2b signaling. fhl1b depletion compromised liver specification and enhanced induction of pancreatic cells from endodermal progenitors. Conversely, overexpression of fhl1b favored liver specification and inhibited induction of pancreatic cells. By single-cell lineage tracing, we showed that fhl1b depletion led lateral endodermal cells, destined to become liver cells, to become pancreatic cells. Reversely, when fhl1b was overexpressed, medially located endodermal cells, fated to differentiate into pancreatic and intestinal cells, contributed to the liver by directly or indirectly modulating the discrete levels of pdx1 expression in endodermal progenitors. Moreover, loss of fhl1b increased the regenerative capacity of β-cells by increasing pdx1 and neurod expression in the hepatopancreatic ductal system. Altogether, these data reveal novel and critical functions of Fhl1b in the hepatic versus pancreatic fate decision and in β-cell regeneration. PMID:26845333
The Simplest Demonstration on Acoustic Beats
ERIC Educational Resources Information Center
Ganci, Alessio; Ganci, Salvatore
2015-01-01
The classical demonstration experiment on acoustic beats using two signal generators and a dual trace oscilloscope is an important ingredient in teaching the subject. This short laboratory note aims to point out what may be the simplest demonstrative experiment on acoustic beats to carry out in a classroom without employing any lab apparatus.
"It's All Human Error!": When a School Science Experiment Fails
ERIC Educational Resources Information Center
Viechnicki, Gail Brendel; Kuipers, Joel
2006-01-01
This paper traces the sophisticated negotiations to re-inscribe the authority of Nature when a school science experiment fails during the enactment of a highly rated science curriculum unit. Drawing on transcriptions from classroom videotapes, we identify and describe four primary patterns of interaction that characterize this process, arguing…
ERIC Educational Resources Information Center
Kehoe, E. James
2006-01-01
The rabbit nictitating membrane (NM) response underwent successive stages of acquisition and extinction training in both delay (Experiment 1) and trace (Experiment 2) classical conditioning. In both cases, successive acquisitions became progressively faster, although the largest, most reliable acceleration occurred between the first and second…
Tracing Attention and the Activation Flow of Spoken Word Planning Using Eye Movements
ERIC Educational Resources Information Center
Roelofs, Ardi
2008-01-01
The flow of activation from concepts to phonological forms within the word production system was examined in 3 experiments. In Experiment 1, participants named pictures while ignoring superimposed distractor pictures that were semantically related, phonologically related, or unrelated. Eye movements and naming latencies were recorded. The…
Promoting Student Metacognition
ERIC Educational Resources Information Center
Tanner, Kimberly D.
2012-01-01
The importance of metacognition in the process of learning is an old idea that can be traced from Socrates' questioning methods to Dewey's twentieth-century stance that one learns more from reflecting on one's experiences than from the actual experiences themselves (Dewey, 1933). What is more recent is the coining of the term "metacognition" and…
Pyo, Dongjin; Hahn, Jong Hoon
2009-01-01
Routine monitoring of microcystin in natural waters is difficult because the concentration of the toxin is usually lower than the detection limits. As a more sensitive detection method for microcystin, we developed a microchip based enzyme-linked immunosorbent assay (ELISA) based on monoclonal antibodies. New monoclonal antibodies against the microcystin leucine-arginine variant (MCLR), a cyclic peptide toxin of the freshwater cyanobacterium Microcystis aeruginosa, were prepared from cloned hybridoma cell lines. We used keyhole limpet hemocyanin(KLH)-conjugated MCLR as an immunogen for the production of mouse monoclonal antibody. The immunization, cell fusion, and screening of hybridoma cells producing anti-MCLR antibody were conducted. Since the ELISA test was highly sensitive, the newly developed microchip based ELISA can be suitable for the trace analysis of cyanobacterial hepatotoxins, microcystins in water. The linear responses of monoclonal antibodies with different concentrations of microcystin LR were established between 0.025 and 0.3 ng/mL.
NASA Technical Reports Server (NTRS)
Vinatier, S.; Bezard, B.; Anderson, C. M.; Coustenis, A.; Teanby, N.
2012-01-01
Titan's northern spring equinox occurred in August 2009. General Circulation Models (e.g. Lebonnois et al., 2012) predict strong modifications of the global circulation in this period, with formation of two circulation cells instead of the pole-to-pole cell that occurred during northern winter. This winter single cell, which had its descending branch at the north pole, was at the origin of the enrichment of molecular abundances and high stratopause temperatures observed by Cassini/CIRS at high northern latitudes (e.g. Achterberg et al., 2011, Coustenis et al., 2010, Teanby et al., 2008, Vinatier et al., 2010). The predicted dynamical seasonal variations after the equinox have strong impact on the spatial distributions of trace gas, temperature and aerosol abundances. We will present here an analysis of CIRS limb-geometry datasets acquired in 2010 and 2011 that we used to monitor the seasonal evolution of the vertical profiles of temperature, molecular (C2H2, C2H6, HCN, ..) and aerosol abundances.
Guernet, Alexis; Mungamuri, Sathish Kumar; Cartier, Dorthe; Sachidanandam, Ravi; Jayaprakash, Anitha; Adriouch, Sahil; Vezain, Myriam; Charbonnier, Françoise; Rohkin, Guy; Coutant, Sophie; Yao, Shen; Ainani, Hassan; Alexandre, David; Tournier, Isabelle; Boyer, Olivier; Aaronson, Stuart A; Anouar, Youssef; Grumolato, Luca
2016-08-04
Intratumor genetic heterogeneity underlies the ability of tumors to evolve and adapt to different environmental conditions. Using CRISPR/Cas9 technology and specific DNA barcodes, we devised a strategy to recapitulate and trace the emergence of subpopulations of cancer cells containing a mutation of interest. We used this approach to model different mechanisms of lung cancer cell resistance to EGFR inhibitors and to assess effects of combined drug therapies. By overcoming intrinsic limitations of current approaches, CRISPR-barcoding also enables investigation of most types of genetic modifications, including repair of oncogenic driver mutations. Finally, we used highly complex barcodes inserted at a specific genome location as a means of simultaneously tracing the fates of many thousands of genetically labeled cancer cells. CRISPR-barcoding is a straightforward and highly flexible method that should greatly facilitate the functional investigation of specific mutations, in a context that closely mimics the complexity of cancer. Copyright © 2016 Elsevier Inc. All rights reserved.
Organic carbon, and not copper, controls denitrification in oxygen minimum zones of the ocean
NASA Astrophysics Data System (ADS)
Ward, Bess B.; Tuit, Caroline B.; Jayakumar, Amal; Rich, Jeremy J.; Moffett, James; Naqvi, S. Wajih A.
2008-12-01
Incubation experiments under trace metal clean conditions and ambient oxygen concentrations were used to investigate the response of microbial assemblages in oxygen minimum zones (OMZs) to additions of organic carbon and copper, two factors that might be expected to limit denitrification in the ocean. In the OMZs of the Eastern Tropical North and South Pacific, denitrification appeared to be limited by organic carbon; exponential cell growth and rapid nitrate and nitrite depletion occurred upon the addition of small amounts of carbon, but copper had no effect. In the OMZ of the Arabian Sea, neither carbon nor copper appeared to be limiting. We hypothesize that denitrification is variable in time and space in the OMZs in ways that may be predictable based on links to the episodic supply of organic substrates from overlying productive surface waters.
Simultaneous cryo X-ray ptychographic and fluorescence microscopy of green algae
DOE Office of Scientific and Technical Information (OSTI.GOV)
Deng, Junjing; Vine, David J.; Chen, Si
Trace metals play important roles in normal and in disease-causing biological functions. X-ray fluorescence microscopy reveals trace elements with no dependence on binding affinities (unlike with visible light fluorophores) and with improved sensitivity relative to electron probes. However, X-ray fluorescence is not very sensitive for showing the light elements that comprise the majority of cellular material. Here we show that X-ray ptychography can be combined with fluorescence to image both cellular structure and trace element distribution in frozen-hydrated cells at cryogenic temperatures, with high structural and chemical fidelity. Ptychographic reconstruction algorithms deliver phase and absorption contrast images at a resolutionmore » beyond that of the illuminating lens or beam size. Using 5.2-keV X-rays, we have obtained sub-30-nm resolution structural images and ~90-nm-resolution fluorescence images of several elements in frozen-hydrated green algae. Finally, this combined approach offers a way to study the role of trace elements in their structural context.« less
Simultaneous cryo X-ray ptychographic and fluorescence microscopy of green algae
Deng, Junjing; Vine, David J.; Chen, Si; ...
2015-02-24
Trace metals play important roles in normal and in disease-causing biological functions. X-ray fluorescence microscopy reveals trace elements with no dependence on binding affinities (unlike with visible light fluorophores) and with improved sensitivity relative to electron probes. However, X-ray fluorescence is not very sensitive for showing the light elements that comprise the majority of cellular material. Here we show that X-ray ptychography can be combined with fluorescence to image both cellular structure and trace element distribution in frozen-hydrated cells at cryogenic temperatures, with high structural and chemical fidelity. Ptychographic reconstruction algorithms deliver phase and absorption contrast images at a resolutionmore » beyond that of the illuminating lens or beam size. Using 5.2-keV X-rays, we have obtained sub-30-nm resolution structural images and ~90-nm-resolution fluorescence images of several elements in frozen-hydrated green algae. Finally, this combined approach offers a way to study the role of trace elements in their structural context.« less
Simultaneous cryo X-ray ptychographic and fluorescence microscopy of green algae
DOE Office of Scientific and Technical Information (OSTI.GOV)
Deng, Junjing; Vine, David J.; Chen, Si
Trace metals play important roles in normal and in disease-causing biological functions. X-ray fluorescence microscopy reveals trace elements with no dependence on binding affinities (unlike with visible light fluorophores) and with improved sensitivity relative to electron probes. However, X-ray fluorescence is not very sensitive for showing the light elements that comprise the majority of cellular material. Here we show that X-ray ptychography can be combined with fluorescence to image both cellular structure and trace element distribution in frozen-hydrated cells at cryogenic temperatures, with high structural and chemical fidelity. Ptychographic reconstruction algorithms deliver phase and absorption contrast images at a resolutionmore » beyond that of the illuminating lens or beam size. Using 5.2-keV X-rays, we have obtained sub-30-nm resolution structural images and similar to 90-nm-resolution fluorescence images of several elements in frozen-hydrated green algae. This combined approach offers a way to study the role of trace elements in their structural context.« less
Dempsey, William P.; Fraser, Scott E.; Pantazis, Periklis
2012-01-01
Background Elucidating the complex cell dynamics (divisions, movement, morphological changes, etc.) underlying embryonic development and adult tissue regeneration requires an efficient means to track cells with high fidelity in space and time. To satisfy this criterion, we developed a transgenic zebrafish line, called PhOTO, that allows photoconvertible optical tracking of nuclear and membrane dynamics in vivo. Methodology PhOTO zebrafish ubiquitously express targeted blue fluorescent protein (FP) Cerulean and photoconvertible FP Dendra2 fusions, allowing for instantaneous, precise targeting and tracking of any number of cells using Dendra2 photoconversion while simultaneously monitoring global cell behavior and morphology. Expression persists through adulthood, making the PhOTO zebrafish an excellent tool for studying tissue regeneration: after tail fin amputation and photoconversion of a ∼100µm stripe along the cut area, marked differences seen in how cells contribute to the new tissue give detailed insight into the dynamic process of regeneration. Photoconverted cells that contributed to the regenerate were separated into three distinct populations corresponding to the extent of cell division 7 days after amputation, and a subset of cells that divided the least were organized into an evenly spaced, linear orientation along the length of the newly regenerating fin. Conclusions/Significance PhOTO zebrafish have wide applicability for lineage tracing at the systems-level in the early embryo as well as in the adult, making them ideal candidate tools for future research in development, traumatic injury and regeneration, cancer progression, and stem cell behavior. PMID:22431986
Analysis of transferred fragrance and its forensic implications.
Gherghel, Simona; Morgan, Ruth M; Blackman, Christopher S; Karu, Kersti; Parkin, Ivan P
2016-12-01
Perfumes are widely used by many people in developed countries, and a large number of both men and women wear perfumes on a daily basis. Analysis of perfume trace materials from clothing is not commonly employed within forensic casework, yet as a form of trace evidence it has the potential to provide valuable intelligence. In order to appreciate the value of trace evidence there is a fundamental need for an evidence base that can both offer insight into how a trace material behaves under different scenarios and activities, and from which inferences can be made. With this purpose a gas chromatography-mass spectrometry method for trace analysis of perfumes was developed. This paper presents two different series of experiments that investigate the dynamics of perfume transfer as a factor of perfume ageing time, and as a factor of perfume contact time. Empirical data showed that both perfume ageing time, and perfume contact time play a key role in the number of perfume components transferred. These studies have implication for forensic protocols, specifically for perfume trace evidence collection, analysis, interpretation, and presentation, and there is potentially great value in analysing perfumes from clothing exhibits in forensic enquiries that involve close contact between individuals, such as sexual assaults. Copyright © 2016 The Authors. Published by Elsevier Ireland Ltd.. All rights reserved.
Fgf10-positive cells represent a progenitor cell population during lung development and postnatally
El Agha, Elie; Herold, Susanne; Alam, Denise Al; Quantius, Jennifer; MacKenzie, BreAnne; Carraro, Gianni; Moiseenko, Alena; Chao, Cho-Ming; Minoo, Parviz; Seeger, Werner; Bellusci, Saverio
2014-01-01
The lung mesenchyme consists of a widely heterogeneous population of cells that play crucial roles during development and homeostasis after birth. These cells belong to myogenic, adipogenic, chondrogenic, neuronal and other lineages. Yet, no clear hierarchy for these lineages has been established. We have previously generated a novel Fgf10iCre knock-in mouse line that allows lineage tracing of Fgf10-positive cells during development and postnatally. Using these mice, we hereby demonstrate the presence of two waves of Fgf10 expression during embryonic lung development: the first wave, comprising Fgf10-positive cells residing in the submesothelial mesenchyme at early pseudoglandular stage (as well as their descendants); and the second wave, comprising Fgf10-positive cells from late pseudoglandular stage (as well as their descendants). Our lineage-tracing data reveal that the first wave contributes to the formation of parabronchial and vascular smooth muscle cells as well as lipofibroblasts at later developmental stages, whereas the second wave does not give rise to smooth muscle cells but to lipofibroblasts as well as an Nkx2.1- E-Cad- Epcam+ Pro-Spc+ lineage that requires further in-depth analysis. During alveologenesis, Fgf10-positive cells give rise to lipofibroblasts rather than alveolar myofibroblasts, and during adult life, a subpopulation of Fgf10-expressing cells represents a pool of resident mesenchymal stromal (stem) cells (MSCs) (Cd45- Cd31- Sca-1+). Taken together, we show for the first time that Fgf10-expressing cells represent a pool of mesenchymal progenitors in the embryonic and postnatal lung. Our findings suggest that Fgf10-positive cells could be useful for developing stem cell-based therapies for treating interstitial lung diseases. PMID:24353064
Yamamoto, Kurumi; Ishimaru, Yoshiro; Ohmoto, Makoto; Matsumoto, Ichiro; Asakura, Tomiko; Abe, Keiko
2011-01-01
Polycystic kidney disease 1-like 3 (Pkd1l3) is expressed specifically in sour-sensing type III taste cells that have synaptic contacts with afferent nerve fibers in circumvallate and foliate papillae located in the posterior region of the tongue, though not in fungiform papillae or the palate. To visualize the gustatory neural pathways that originate from type III taste cells in circumvallate and foliate papillae, we established transgenic mouse lines that express the transneuronal tracer wheat germ agglutinin (WGA) under the control of the mouse Pkd1l3 gene promoter/enhancer. The WGA transgene was accurately expressed in Pkd1l3-expressing type III taste cells in circumvallate and foliate papillae. Punctate WGA protein signals appeared to be detected specifically in type III taste cells but not in other types of taste cells. WGA protein was transferred primarily to a subset of neurons located in close proximity to the glossopharyngeal nerve bundles in the nodose/petrosal ganglion. WGA signals were also observed in a small population of neurons in the geniculate ganglion. This result demonstrates the anatomical connection between taste receptor cells in the foliate papillae and the chorda tympani nerves. WGA protein was further conveyed to neurons in a rostro-central subdivision of the nucleus of the solitary tract. These findings demonstrate that the approximately 10 kb 5’-flanking region of the mouse Pkd1l3 gene functions as a type III taste cell-specific promoter/enhancer. In addition, experiments using the pkd1l3-WGA transgenic mice reveal a sour gustatory pathway that originates from taste receptor cells in the posterior region of the tongue. PMID:21883212
The existence of adult pancreatic progenitor cells has been debated. While some favor the concept of facultative progenitors involved in homeostasis and repair, neither a location nor markers for such cells have been defined. Using genetic lineage tracing, we show that Doublecortin-like kinase-1 (Dclk1) labels a rare population of long-lived, quiescent pancreatic cells. In vitro, Dclk1+ cells proliferate readily and sustain pancreatic organoid growth. In vivo, Dclk1+ cells are necessary for pancreatic regeneration following injury and chronic inflammation.
ROLE OF NEUROTRANSMITTERS AND PROTEIN SYNTHESIS IN SHORT- AND LONG-TERM MEMORY
DOE Office of Scientific and Technical Information (OSTI.GOV)
Bennett, E.L.; Rosenzweig, M.R.; Flood, J.F.
1978-10-01
Anisomycin is an effective inhibitor of cerebral protein synthesis in mice and is also an effective amnestic agent for both passive and active behavioral tasks. From use of anisomycin in combination with a variety of stimulant and depressant drugs, we conclude that the level of arousal following acquisition plays an important role in determining the duration and the rate of the biosynthetic phase of memory formation. While we have interpreted the experiments with anisomycin as evidence for an essential role of protein in memory storage, others have suggested that side effects of inhibitors of protein synthesis on catecholamine metabolism aremore » the main cause of amnesia. Several experiments were therefore done to compare the effects of anisemycin and catecholamine inhibitors on memory. We conclude that anisomycin's principal amnestic mechanism does not involve inhibition of the catecholamine system. The results strengthen our conclusion that protein synthesis is an essential component for longterm memory trace formation. Also, it is suggested that proteins synthesized in the neuronal cell body are used, in conjunction with other molecules, to produce permanent and semi-permanent anatomical changes.« less
Multi -omics and metabolic modelling pipelines: challenges and tools for systems microbiology.
Fondi, Marco; Liò, Pietro
2015-02-01
Integrated -omics approaches are quickly spreading across microbiology research labs, leading to (i) the possibility of detecting previously hidden features of microbial cells like multi-scale spatial organization and (ii) tracing molecular components across multiple cellular functional states. This promises to reduce the knowledge gap between genotype and phenotype and poses new challenges for computational microbiologists. We underline how the capability to unravel the complexity of microbial life will strongly depend on the integration of the huge and diverse amount of information that can be derived today from -omics experiments. In this work, we present opportunities and challenges of multi -omics data integration in current systems biology pipelines. We here discuss which layers of biological information are important for biotechnological and clinical purposes, with a special focus on bacterial metabolism and modelling procedures. A general review of the most recent computational tools for performing large-scale datasets integration is also presented, together with a possible framework to guide the design of systems biology experiments by microbiologists. Copyright © 2015. Published by Elsevier GmbH.
Arni, Caroline
2008-01-01
As an experimental medical practice artificial insemination in humans dates back to the end of the 18th century. Efforts intensified in the second half of the 19th century, when, especially in France, the number of reports in scientific publications increased and the topic became the subject of heated debates. I trace this emergence of reproductive medicine avant la lettre by reviewing the relevant medical publications. Hereby, I focus on how experiments in artificial insemination presupposed a new conceptualisation of procreation which detaches procreation from the doings of human actors by reducing it to the fusion of germ cells. This "biologisation" of procreation entailed a series of irritations with regard to the determination of "natural" paternity and the impact and relevance of the procreative act's nature. These irritations are dealt with in a novel that appears in Paris in 1884, entitled The Man-Maker. This novel thus attests to the cultural uncertainties that went along with the emergence of what laid the epistemological ground for the reproductive medicine to come.
NASA Astrophysics Data System (ADS)
Patterson, B. M.; Shackleton, M.; Furness, A. J.; Bekele, E.; Pearce, J.; Linge, K. L.; Busetti, F.; Spadek, T.; Toze, S.
2011-03-01
The fate of nine trace organic compounds was evaluated during a 12 month large-scale laboratory column experiment. The columns were packed with aquifer sediment and evaluated under natural aerobic and artificial anaerobic geochemical conditions, to assess the potential for natural attenuation of these compounds during aquifer passage associated with managed aquifer recharge (MAR). The nine trace organic compounds were bisphenol A (BPA), 17β-estradiol (E2), 17α-ethynylestradiol (EE2), N-nitrosodimethylamine (NDMA), N-nitrosomorpholine (NMOR), carbamazepine, oxazepam, iohexol and iodipamide. In the low organic carbon content Spearwood sediment, all trace organics were non-retarded with retardation coefficients between 1.0 and 1.2, indicating that these compounds would travel at near groundwater velocities within the aquifer. The natural aerobic geochemical conditions provided a suitable environment for the rapid degradation for BPA, E2, iohexol (half life < 1 day). Lag-times for the start of degradation of these compounds ranged from < 15 to 30 days. While iodipamide was persistent under aerobic conditions, artificial reductive geochemical conditions promoted via the addition of ethanol, resulted in rapid degradation (half life < 1 days). Pharmaceuticals (carbamazepine and oxazepam) and disinfection by-products (NDMA and NMOR) did not degrade under either aerobic or anaerobic aquifer geochemical conditions (half life > 50 days). Field-based validation experiments with carbamazepine and oxazepam also showed no degradation. If persistent trace organics are present in recycled waters at concentrations in excess of their intended use, natural attenuation during aquifer passage alone may not result in extracted water meeting regulatory requirements. Additional pre treatment of the recycled water would therefore be required.
Single-cell computational analysis of light harvesting in a flat-panel photo-bioreactor.
Loomba, Varun; Huber, Gregor; von Lieres, Eric
2018-01-01
Flat-panel photo-bioreactors (PBRs) are customarily applied for investigating growth of microalgae. Optimal design and operation of such reactors is still a challenge due to complex non-linear combinations of various impact factors, particularly hydrodynamics, light irradiation, and cell metabolism. A detailed analysis of single-cell light reception can lead to novel insights into the complex interactions of light exposure and algae movement in the reactor. The combined impacts of hydrodynamics and light irradiation on algae cultivation in a flat-panel PBR were studied by tracing the light exposure of individual cells over time. Hydrodynamics and turbulent mixing in this air-sparged bioreactor were simulated using the Eulerian approach for the liquid phase and a slip model for the gas phase velocity profiles. The liquid velocity was then used for tracing single cells and their light exposure, using light intensity profiles obtained from solving the radiative transfer equation at different wavelengths. The residence times of algae cells in defined dark and light zones of the PBR were statistically analyzed for different algal concentrations and sparging rates. The results indicate poor mixing caused by the reactor design which can be only partially improved by increased sparging rates. The results provide important information for optimizing algal biomass productivity by improving bioreactor design and operation and can further be utilized for an in-depth analysis of algal growth by using advanced models of cell metabolism.
Infant word recognition: Insights from TRACE simulations☆
Mayor, Julien; Plunkett, Kim
2014-01-01
The TRACE model of speech perception (McClelland & Elman, 1986) is used to simulate results from the infant word recognition literature, to provide a unified, theoretical framework for interpreting these findings. In a first set of simulations, we demonstrate how TRACE can reconcile apparently conflicting findings suggesting, on the one hand, that consonants play a pre-eminent role in lexical acquisition (Nespor, Peña & Mehler, 2003; Nazzi, 2005), and on the other, that there is a symmetry in infant sensitivity to vowel and consonant mispronunciations of familiar words (Mani & Plunkett, 2007). In a second series of simulations, we use TRACE to simulate infants’ graded sensitivity to mispronunciations of familiar words as reported by White and Morgan (2008). An unexpected outcome is that TRACE fails to demonstrate graded sensitivity for White and Morgan’s stimuli unless the inhibitory parameters in TRACE are substantially reduced. We explore the ramifications of this finding for theories of lexical development. Finally, TRACE mimics the impact of phonological neighbourhoods on early word learning reported by Swingley and Aslin (2007). TRACE offers an alternative explanation of these findings in terms of mispronunciations of lexical items rather than imputing word learning to infants. Together these simulations provide an evaluation of Developmental (Jusczyk, 1993) and Familiarity (Metsala, 1999) accounts of word recognition by infants and young children. The findings point to a role for both theoretical approaches whereby vocabulary structure and content constrain infant word recognition in an experience-dependent fashion, and highlight the continuity in the processes and representations involved in lexical development during the second year of life. PMID:24493907
Infant word recognition: Insights from TRACE simulations.
Mayor, Julien; Plunkett, Kim
2014-02-01
The TRACE model of speech perception (McClelland & Elman, 1986) is used to simulate results from the infant word recognition literature, to provide a unified, theoretical framework for interpreting these findings. In a first set of simulations, we demonstrate how TRACE can reconcile apparently conflicting findings suggesting, on the one hand, that consonants play a pre-eminent role in lexical acquisition (Nespor, Peña & Mehler, 2003; Nazzi, 2005), and on the other, that there is a symmetry in infant sensitivity to vowel and consonant mispronunciations of familiar words (Mani & Plunkett, 2007). In a second series of simulations, we use TRACE to simulate infants' graded sensitivity to mispronunciations of familiar words as reported by White and Morgan (2008). An unexpected outcome is that TRACE fails to demonstrate graded sensitivity for White and Morgan's stimuli unless the inhibitory parameters in TRACE are substantially reduced. We explore the ramifications of this finding for theories of lexical development. Finally, TRACE mimics the impact of phonological neighbourhoods on early word learning reported by Swingley and Aslin (2007). TRACE offers an alternative explanation of these findings in terms of mispronunciations of lexical items rather than imputing word learning to infants. Together these simulations provide an evaluation of Developmental (Jusczyk, 1993) and Familiarity (Metsala, 1999) accounts of word recognition by infants and young children. The findings point to a role for both theoretical approaches whereby vocabulary structure and content constrain infant word recognition in an experience-dependent fashion, and highlight the continuity in the processes and representations involved in lexical development during the second year of life.
A finite volume method for trace element diffusion and partitioning during crystal growth
NASA Astrophysics Data System (ADS)
Hesse, Marc A.
2012-09-01
A finite volume method on a uniform grid is presented to compute the polythermal diffusion and partitioning of a trace element during the growth of a porphyroblast crystal in a uniform matrix and in linear, cylindrical and spherical geometry. The motion of the crystal-matrix interface and the thermal evolution are prescribed functions of time. The motion of the interface is discretized and it advances from one cell boundary to next as the prescribed interface position passes the cell center. The appropriate conditions for the flux across the crystal-matrix interface are derived from discrete mass conservation. Numerical results are benchmarked against steady and transient analytic solutions for isothermal diffusion with partitioning and growth. Two applications illustrate the ability of the model to reproduce observed rare-earth element patterns in garnets (Skora et al., 2006) and water concentration profiles around spherulites in obsidian (Watkins et al., 2009). Simulations with diffusion inside the growing crystal show complex concentration evolutions for trace elements with high diffusion coefficients, such as argon or hydrogen, but demonstrate that rare-earth element concentrations in typical metamorphic garnets are not affected by intracrystalline diffusion.
Use of diluted urine for cultivation of Chlorella vulgaris.
Jaatinen, Sanna; Lakaniemi, Aino-Maija; Rintala, Jukka
2016-01-01
Our aim was to study the biomass growth of microalga Chlorella vulgaris using diluted human urine as a sole nutrient source. Batch cultivations (21 days) were conducted in five different urine dilutions (1:25-1:300), in 1:100-diluted urine as such and with added trace elements, and as a reference, in artificial growth medium. The highest biomass density was obtained in 1:100-diluted urine with and without additional trace elements (0.73 and 0.60 g L(-1), respectively). Similar biomass growth trends and densities were obtained with 1:25- and 1:300-diluted urine (0.52 vs. 0.48 gVSS L(-1)) indicating that urine at dilution 1:25 can be used to cultivate microalgal based biomass. Interestingly, even 1:300-diluted urine contained sufficiently nutrients and trace elements to support biomass growth. Biomass production was similar despite pH-variation from < 5 to 9 in different incubations indicating robustness of the biomass growth. Ammonium formation did not inhibit overall biomass growth. At the beginning of cultivation, the majority of the biomass consisted of living algal cells, while towards the end, their share decreased and the estimated share of bacteria and cell debris increased.