Sample records for slice cultures developmental

  1. The mouse cerebellar cortex in organotypic slice cultures: an in vitro model to analyze the consequences of mutations and pathologies on neuronal survival, development, and function.

    PubMed

    Lonchamp, Etienne; Dupont, Jean-Luc; Beekenkamp, Huguette; Poulain, Bernard; Bossu, Jean-Louis

    2006-01-01

    Thin acute slices and dissociated cell cultures taken from different parts of the brain have been widely used to examine the function of the nervous system, neuron-specific interactions, and neuronal development (specifically, neurobiology, neuropharmacology, and neurotoxicology studies). Here, we focus on an alternative in vitro model: brain-slice cultures in roller tubes, initially introduced by Beat Gähwiler for studies with rats, that we have recently adapted for studies of mouse cerebellum. Cultured cerebellar slices afford many of the advantages of dissociated cultures of neurons and thin acute slices. Organotypic slice cultures were established from newborn or 10-15-day-old mice. After 3-4 weeks in culture, the slices flattened to form a cell monolayer. The main types of cerebellar neurons could be identified with immunostaining techniques, while their electrophysiological properties could be easily characterized with the patch-clamp recording technique. When slices were taken from newborn mice and cultured for 3 weeks, aspects of the cerebellar development were displayed. A functional neuronal network was established despite the absence of mossy and climbing fibers, which are the two excitatory afferent projections to the cerebellum. When slices were made from 10-15-day-old mice, which are at a developmental stage when cerebellum organization is almost established, the structure and neuronal pathways were intact after 3-4 weeks in culture. These unique characteristics make organotypic slice cultures of mouse cerebellar cortex a valuable model for analyzing the consequences of gene mutations that profoundly alter neuronal function and compromise postnatal survival.

  2. Developmental excitatory-to-inhibitory GABA polarity switch is delayed in Ts65Dn mice, a genetic model of Down syndrome.

    PubMed

    Lysenko, Larisa V; Kim, Jeesun; Madamba, Francisco; Tyrtyshnaia, Anna A; Ruparelia, Aarti; Kleschevnikov, Alexander M

    2018-07-01

    Down syndrome (DS) is the most frequent genetic cause of developmental abnormalities leading to intellectual disability. One notable phenomenon affecting the formation of nascent neural circuits during late developmental periods is developmental switch of GABA action from depolarizing to hyperpolarizing mode. We examined properties of this switch in DS using primary cultures and acute hippocampal slices from Ts65Dn mice, a genetic model of DS. Cultures of DIV3-DIV13 Ts65Dn and control normosomic (2 N) neurons were loaded with FURA-2 AM, and GABA action was assessed using local applications. In 2 N cultures, the number of GABA-activated cells dropped from ~100% to 20% between postnatal days 3-13 (P3-P13) reflecting the switch in GABA action polarity. In Ts65Dn cultures, the timing of this switch was delayed by 2-3 days. Next, microelectrode recordings of multi-unit activity (MUA) were performed in CA3 slices during bath application of the GABA A agonist isoguvacine. MUA frequency was increased in P8-P12 and reduced in P14-P22 slices reflecting the switch of GABA action from excitatory to inhibitory mode. The timing of this switch was delayed in Ts65Dn by approximately 2 days. Finally, frequency of giant depolarizing potentials (GDPs), a form of primordial neural activity, was significantly increased in slices from Ts65Dn pups at P12 and P14. These experimental evidences show that GABA action polarity switch is delayed in Ts65Dn model of DS, and that these changes lead to a delay in maturation of nascent neural circuits. These alterations may affect properties of neural circuits in adult animals and, therefore, represent a prospective target for pharmacotherapy of cognitive impairment in DS. Copyright © 2018 Elsevier Inc. All rights reserved.

  3. Free-floating adult human brain-derived slice cultures as a model to study the neuronal impact of Alzheimer's disease-associated Aβ oligomers.

    PubMed

    Mendes, Niele D; Fernandes, Artur; Almeida, Glaucia M; Santos, Luis E; Selles, Maria Clara; Lyra-Silva, Natalia; Machado, Carla M; Horta-Júnior, José A C; Louzada, Paulo R; De Felice, Fernanda G; Alvez-Leon, Soniza; Marcondes, Jorge; Assirati, João Alberto; Matias, Caio M; Klein, William L; Garcia-Cairasco, Norberto; Ferreira, Sergio T; Neder, Luciano; Sebollela, Adriano

    2018-05-31

    Slice cultures have been prepared from several organs. With respect to the brain, advantages of slice cultures over dissociated cell cultures include maintenance of the cytoarchitecture and neuronal connectivity. Slice cultures from adult human brain have been reported and constitute a promising method to study neurological diseases. Despite this potential, few studies have characterized in detail cell survival and function along time in short-term, free-floating cultures. We used tissue from adult human brain cortex from patients undergoing temporal lobectomy to prepare 200 μm-thick slices. Along the period in culture, we evaluated neuronal survival, histological modifications, and neurotransmitter release. The toxicity of Alzheimer's-associated Aβ oligomers (AβOs) to cultured slices was also analyzed. Neurons in human brain slices remain viable and neurochemically active for at least four days in vitro, which allowed detection of binding of AβOs. We further found that slices exposed to AβOs presented elevated levels of hyperphosphorylated Tau, a hallmark of Alzheimer's disease. Although slice cultures from adult human brain have been previously prepared, this is the first report to analyze cell viability and neuronal activity in short-term free-floating cultures as a function of days in vitro. Once surgical tissue is available, the current protocol is easy to perform and produces functional slices from adult human brain. These slice cultures may represent a preferred model for translational studies of neurodegenerative disorders when long term culturing in not required, as in investigations on AβO neurotoxicity. Copyright © 2018 Elsevier B.V. All rights reserved.

  4. Rat brain sagittal organotypic slice cultures as an ex vivo dopamine cell loss system.

    PubMed

    McCaughey-Chapman, Amy; Connor, Bronwen

    2017-02-01

    Organotypic brain slice cultures are a useful tool to study neurological function as they provide a more complex, 3-dimensional system than standard 2-dimensional in vitro cell cultures. Building on a previously developed mouse brain slice culture protocol, we have developed a rat sagittal brain slice culture system as an ex vivo model of dopamine cell loss. We show that rat brain organotypic slice cultures remain viable for up to 6 weeks in culture. Using Fluoro-Gold axonal tracing, we demonstrate that the slice 3-dimensional cytoarchitecture is maintained over a 4 week culturing period, with particular focus on the nigrostriatal pathway. Treatment of the cultures with 6-hydroxydopamine and desipramine induces a progressive loss of Fluoro-Gold-positive nigral cells with a sustained loss of tyrosine hydroxylase-positive nigral cells. This recapitulates the pattern of dopaminergic degeneration observed in the rat partial 6-hydroxydopamine lesion model and, most importantly, the progressive pathology of Parkinson's disease. Our slice culture platform provides an advance over other systems, as we demonstrate for the first time 3-dimensional cytoarchitecture maintenance of rat nigrostriatal sagittal slices for up to 6 weeks. Our ex vivo organotypic slice culture system provides a long term cellular platform to model Parkinson's disease, allowing for the elucidation of mechanisms involved in dopaminergic neuron degeneration and the capability to study cellular integration and plasticity ex vivo. Copyright © 2017 Elsevier B.V. All rights reserved.

  5. Trajectory Analysis Unveils Reelin's Role in the Directed Migration of Granule Cells in the Dentate Gyrus.

    PubMed

    Wang, Shaobo; Brunne, Bianka; Zhao, Shanting; Chai, Xuejun; Li, Jiawei; Lau, Jeremie; Failla, Antonio Virgilio; Zobiak, Bernd; Sibbe, Mirjam; Westbrook, Gary L; Lutz, David; Frotscher, Michael

    2018-01-03

    Reelin controls neuronal migration and layer formation. Previous studies in reeler mice deficient in Reelin focused on the result of the developmental process in fixed tissue sections. It has remained unclear whether Reelin affects the migratory process, migration directionality, or migrating neurons guided by the radial glial scaffold. Moreover, Reelin has been regarded as an attractive signal because newly generated neurons migrate toward the Reelin-containing marginal zone. Conversely, Reelin might be a stop signal because migrating neurons in reeler , but not in wild-type mice, invade the marginal zone. Here, we monitored the migration of newly generated proopiomelanocortin-EGFP -expressing dentate granule cells in slice cultures from reeler , reeler -like mutants and wild-type mice of either sex using real-time microscopy. We discovered that not the actual migratory process and migratory speed, but migration directionality of the granule cells is controlled by Reelin. While wild-type granule cells migrated toward the marginal zone of the dentate gyrus, neurons in cultures from reeler and reeler -like mutants migrated randomly in all directions as revealed by vector analyses of migratory trajectories. Moreover, live imaging of granule cells in reeler slices cocultured to wild-type dentate gyrus showed that the reeler neurons changed their directions and migrated toward the Reelin-containing marginal zone of the wild-type culture, thus forming a compact granule cell layer. In contrast, directed migration was not observed when Reelin was ubiquitously present in the medium of reeler slices. These results indicate that topographically administered Reelin controls the formation of a granule cell layer. SIGNIFICANCE STATEMENT Neuronal migration and the various factors controlling its onset, speed, directionality, and arrest are poorly understood. Slice cultures offer a unique model to study the migration of individual neurons in an almost natural environment. In the present study, we took advantage of the expression of proopiomelanocortin-EGFP by newly generated, migrating granule cells to analyze their migratory trajectories in hippocampal slice cultures from wild-type mice and mutants deficient in Reelin signaling. We show that the compartmentalized presence of Reelin is essential for the directionality, but not the actual migratory process or speed, of migrating granule cells leading to their characteristic lamination in the dentate gyrus. Copyright © 2018 the authors 0270-6474/18/380137-12$15.00/0.

  6. Organotypic Slice Cultures for Studies of Postnatal Neurogenesis

    PubMed Central

    Mosa, Adam J.; Wang, Sabrina; Tan, Yao Fang; Wojtowicz, J. Martin

    2015-01-01

    Here we describe a technique for studying hippocampal postnatal neurogenesis in the rodent brain using the organotypic slice culture technique. This method maintains the characteristic topographical morphology of the hippocampus while allowing direct application of pharmacological agents to the developing hippocampal dentate gyrus. Additionally, slice cultures can be maintained for up to 4 weeks and thus, allow one to study the maturation process of newborn granule neurons. Slice cultures allow for efficient pharmacological manipulation of hippocampal slices while excluding complex variables such as uncertainties related to the deep anatomic location of the hippocampus as well as the blood brain barrier. For these reasons, we sought to optimize organotypic slice cultures specifically for postnatal neurogenesis research. PMID:25867138

  7. The Utility of Thin Slice Ratings for Predicting Language Growth in Children with Autism Spectrum Disorder

    ERIC Educational Resources Information Center

    Walton, Katherine M.; Ingersoll, Brooke R.

    2016-01-01

    Literature on "Thin Slice" ratings indicates that a number of personality characteristics and behaviors can be accurately predicted by ratings of very short segments (<5?min) of behavior. This study examined the utility of Thin Slice ratings of young children with autism spectrum disorder for predicting developmental skills and…

  8. A simple method for multiday imaging of slice cultures.

    PubMed

    Seidl, Armin H; Rubel, Edwin W

    2010-01-01

    The organotypic slice culture (Stoppini et al. A simple method for organotypic cultures of nervous tissue. 1991;37:173-182) has become the method of choice to answer a variety of questions in neuroscience. For many experiments, however, it would be beneficial to image or manipulate a slice culture repeatedly, for example, over the course of many days. We prepared organotypic slice cultures of the auditory brainstem of P3 and P4 mice and kept them in vitro for up to 4 weeks. Single cells in the auditory brainstem were transfected with plasmids expressing fluorescent proteins by way of electroporation (Haas et al. Single-cell electroporation for gene transfer in vivo. 2001;29:583-591). The culture was then placed in a chamber perfused with oxygenated ACSF and the labeled cell imaged with an inverted wide-field microscope repeatedly for multiple days, recording several time-points per day, before returning the slice to the incubator. We describe a simple method to image a slice culture preparation during the course of multiple days and over many continuous hours, without noticeable damage to the tissue or photobleaching. Our method uses a simple, inexpensive custom-built insulator constructed around the microscope to maintain controlled temperature and uses a perfusion chamber as used for in vitro slice recordings. (c) 2009 Wiley-Liss, Inc.

  9. Organotypic slice cultures containing the preBötzinger complex generate respiratory-like rhythms

    PubMed Central

    Phillips, Wiktor S.; Herly, Mikkel; Del Negro, Christopher A.

    2015-01-01

    Study of acute brain stem slice preparations in vitro has advanced our understanding of the cellular and synaptic mechanisms of respiratory rhythm generation, but their inherent limitations preclude long-term manipulation and recording experiments. In the current study, we have developed an organotypic slice culture preparation containing the preBötzinger complex (preBötC), the core inspiratory rhythm generator of the ventrolateral brain stem. We measured bilateral synchronous network oscillations, using calcium-sensitive fluorescent dyes, in both ventrolateral (presumably the preBötC) and dorsomedial regions of slice cultures at 7–43 days in vitro. These calcium oscillations appear to be driven by periodic bursts of inspiratory neuronal activity, because whole cell recordings from ventrolateral neurons in culture revealed inspiratory-like drive potentials, and no oscillatory activity was detected from glial fibrillary associated protein-expressing astrocytes in cultures. Acute slices showed a burst frequency of 10.9 ± 4.2 bursts/min, which was not different from that of brain stem slice cultures (13.7 ± 10.6 bursts/min). However, slice cocultures that include two cerebellar explants placed along the dorsolateral border of the brainstem displayed up to 193% faster burst frequency (22.4 ± 8.3 bursts/min) and higher signal amplitude (340%) compared with acute slices. We conclude that preBötC-containing slice cultures retain inspiratory-like rhythmic function and therefore may facilitate lines of experimentation that involve extended incubation (e.g., genetic transfection or chronic drug exposure) while simultaneously being amenable to imaging and electrophysiology at cellular, synaptic, and network levels. PMID:26655824

  10. Metabolism of 2,2′,3,3′,6,6′-Hexachlorobiphenyl (PCB 136) Atropisomers in Tissue Slices from Phenobarbital or Dexamethasone-Induced Rats is Sex-Dependent

    PubMed Central

    Wu, Xianai; Kania-Korwel, Izabela; Chen, Hao; Stamou, Marianna; Dammanahalli, Karigowda J.; Duffel, Michael; Lein, Pamela J.; Lehmler, Hans-Joachim

    2013-01-01

    Chiral polychlorinated biphenyls (PCBs) such as PCB 136 enantioselectively sensitize the ryanodine receptor (RyR). In light of recent evidence that PCBs cause developmental neurotoxicity via RyR-dependent mechanisms, this suggests that enantioselective PCB metabolism may influence the developmental neurotoxicity of chiral PCBs. However, enantioselective disposition of PCBs has not been fully characterized.The effect of sex and cytochrome P450 (P450) enzyme induction on the enantioselective metabolism of PCB 136 was studied using liver tissue slices prepared from naïve control (CTL), phenobarbital (PB; CYP2B inducer) or dexamethasone (DEX; CYP3A inducer) pretreated adult Sprague-Dawley rats. PCB 136 metabolism was also examined in hippocampal slices derived from untreated rat pups.In liver tissue slices, hydroxylated PCB (OH-PCB) profiles depended on sex and inducer pretreatment, and OH-PCB levels followed the rank orders male > female and PB > DEX > CTL. In contrast, the enantiomeric enrichment of PCB 136 and its metabolites was independent of sex and inducer pretreatment. Only small amounts of PCB 136 partitioned into hippocampal tissue slices and no OH-PCB metabolites were detected.Our results suggest that enantioselective metabolism, sex and induction status of P450 enzymes in the liver may modulate the neurotoxic outcomes of developmental exposure to chiral PCBs. PMID:23581876

  11. Developmental imaging: the avian embryo hatches to the challenge.

    PubMed

    Kulesa, Paul M; McKinney, Mary C; McLennan, Rebecca

    2013-06-01

    The avian embryo provides a multifaceted model to study developmental mechanisms because of its accessibility to microsurgery, fluorescence cell labeling, in vivo imaging, and molecular manipulation. Early two-dimensional planar growth of the avian embryo mimics human development and provides unique access to complex cell migration patterns using light microscopy. Later developmental events continue to permit access to both light and other imaging modalities, making the avian embryo an excellent model for developmental imaging. For example, significant insights into cell and tissue behaviors within the primitive streak, craniofacial region, and cardiovascular and peripheral nervous systems have come from avian embryo studies. In this review, we provide an update to recent advances in embryo and tissue slice culture and imaging, fluorescence cell labeling, and gene profiling. We focus on how technical advances in the chick and quail provide a clearer understanding of how embryonic cell dynamics are beautifully choreographed in space and time to sculpt cells into functioning structures. We summarize how these technical advances help us to better understand basic developmental mechanisms that may lead to clinical research into human birth defects and tissue repair. Copyright © 2013 Wiley Periodicals, Inc.

  12. Acute Hippocampal Slice Preparation and Hippocampal Slice Cultures

    PubMed Central

    Lein, Pamela J.; Barnhart, Christopher D.; Pessah, Isaac N.

    2012-01-01

    A major advantage of hippocampal slice preparations is that the cytoarchitecture and synaptic circuits of the hippocampus are largely retained. In neurotoxicology research, organotypic hippocampal slices have mostly been used as acute ex vivo preparations for investigating the effects of neurotoxic chemicals on synaptic function. More recently, hippocampal slice cultures, which can be maintained for several weeks to several months in vitro, have been employed to study how neurotoxic chemicals influence the structural and functional plasticity in hippocampal neurons. This chapter provides protocols for preparing hippocampal slices to be used acutely for electrophysiological measurements using glass microelectrodes or microelectrode arrays or to be cultured for morphometric assessments of individual neurons labeled using biolistics. PMID:21815062

  13. Three-dimensional slice cultures from murine fetal gut for investigations of the enteric nervous system.

    PubMed

    Metzger, Marco; Bareiss, Petra M; Nikolov, Ivan; Skutella, Thomas; Just, Lothar

    2007-01-01

    Three-dimensional intestinal cultures offer new possibilities for the examination of growth potential, analysis of time specific gene expression, and spatial cellular arrangement of enteric nervous system in an organotypical environment. We present an easy to produce in vitro model of the enteric nervous system for analysis and manipulation of cellular differentiation processes. Slice cultures of murine fetal colon were cultured on membrane inserts for up to 2 weeks without loss of autonomous contractility. After slice preparation, cultured tissue reorganized within the first days in vitro. Afterward, the culture possessed more than 35 cell layers, including high prismatic epithelial cells, smooth muscle cells, glial cells, and neurons analyzed by immunohistochemistry. The contraction frequency of intestinal slice culture could be modulated by the neurotransmitter serotonin and the sodium channel blocker tetrodotoxin. Coculture experiments with cultured neurospheres isolated from enhanced green fluorescent protein (eGFP) transgenic mice demonstrated that differentiating eGFP-positive neurons were integrated into the intestinal tissue culture. This slice culture model of enteric nervous system proved to be useful for studying cell-cell interactions, cellular signaling, and cell differentiation processes in a three-dimensional cell arrangement.

  14. Organotypic hippocampal slice culture from the adult mouse brain: a versatile tool for translational neuropsychopharmacology.

    PubMed

    Kim, Hyunjeong; Kim, Eosu; Park, Minsun; Lee, Eun; Namkoong, Kee

    2013-03-05

    One of the most significant barriers towards translational neuropsychiatry would be an unavailability of living brain tissues. Although organotypic brain tissue culture could be a useful alternative enabling observation of temporal changes induced by various drugs in living brain tissues, a proper method to establish a stable organotypic brain slice culture system using adult (rather than neonatal) hippocampus has been still elusive. In this study, we evaluated our simple method using the serum-free culture medium for successful adult organotypic hippocampal slice culture. Several tens of hippocampal slices from a single adult mouse (3-5 months old) were cultured in serum-free versus serum-containing conventional culture medium for 30 days and underwent various experiments to validate the effects of the existence of serum in the culture medium. Neither the excessive regression of neuronal viability nor metabolic deficiency was observed in the serum-free medium culture in contrast to the serum-containing medium culture. Despite such viability, newly generated immature neurons were scarcely detected in the serum-free culture, suggesting that the original neurons in the brain slice persist rather than being replaced by neurogenesis. Key structural features of in vivo neural tissue constituting astrocytes, neural processes, and pre- and post-synapses were also well preserved in the serum-free culture. In conclusion, using the serum-free culture medium, the adult hippocampal slice culture system will serve as a promising ex vivo tool for various fields of neuroscience, especially for studies on aging-related neuropsychiatric disorders or for high throughput screening of potential agents working against such disorders. Copyright © 2012 Elsevier Inc. All rights reserved.

  15. Characterization of cortical neuronal and glial alterations during culture of organotypic whole brain slices from neonatal and mature mice.

    PubMed

    Staal, Jerome A; Alexander, Samuel R; Liu, Yao; Dickson, Tracey D; Vickers, James C

    2011-01-01

    Organotypic brain slice culturing techniques are extensively used in a wide range of experimental procedures and are particularly useful in providing mechanistic insights into neurological disorders or injury. The cellular and morphological alterations associated with hippocampal brain slice cultures has been well established, however, the neuronal response of mouse cortical neurons to culture is not well documented. In the current study, we compared the cell viability, as well as phenotypic and protein expression changes in cortical neurons, in whole brain slice cultures from mouse neonates (P4-6), adolescent animals (P25-28) and mature adults (P50+). Cultures were prepared using the membrane interface method. Propidium iodide labeling of nuclei (due to compromised cell membrane) and AlamarBlue™ (cell respiration) analysis demonstrated that neonatal tissue was significantly less vulnerable to long-term culture in comparison to the more mature brain tissues. Cultures from P6 animals showed a significant increase in the expression of synaptic markers and a decrease in growth-associated proteins over the entire culture period. However, morphological analysis of organotypic brain slices cultured from neonatal tissue demonstrated that there were substantial changes to neuronal and glial organization within the neocortex, with a distinct loss of cytoarchitectural stratification and increased GFAP expression (p<0.05). Additionally, cultures from neonatal tissue had no glial limitans and, after 14 DIV, displayed substantial cellular protrusions from slice edges, including cells that expressed both glial and neuronal markers. In summary, we present a substantial evaluation of the viability and morphological changes that occur in the neocortex of whole brain tissue cultures, from different ages, over an extended period of culture.

  16. Development and characterization of an ex-vivo brain slice culture model of chronic wasting disease

    USDA-ARS?s Scientific Manuscript database

    Prion diseases have long incubation times in vivo, therefore, modeling the diseases ex-vivo will advance the development of rationale-based therapeutic strategies. An organotypic slice culture assay (POSCA) was recently developed for scrapie prions by inoculating mouse cerebellar brain slices with R...

  17. Effects of neural stem cell media on hypoxic injury in rat hippocampal slice cultures.

    PubMed

    Lee, Na Mi; Chae, Soo Ahn; Lee, Hong Jun

    2017-12-15

    Neonatal hypoxic-ischemic brain injuries cause serious neurological sequelae, yet there is currently no effective treatment for them. We hypothesized that neurotrophic factors released into the medium by stem cells could supply hypoxia-damaged organotypic hippocampal slice cultures with regenerative abilities. We prepared organotypic slice cultures of the hippocampus of 7-day-old Sprague-Dawley rats based on the modified Stoppini method; slices were cultured for 14days in vitro using either Gahwiler's medium (G-medium) or stem cell-conditioned medium (S-medium) as culture medium. At day 14 in vitro, hippocampal slice cultures were exposed to 95% N 2 and 5% CO 2 for 3h to induce hypoxic damage, the extent of which was then measured using propidium iodide fluorescence and immunohistochemistry images. We performed dot blotting to estimate neurotrophic/growth factor levels in the G- and S-media. Organotypic hippocampal slices cultured using S-medium after hypoxic injury were significantly less damaged than those cultured using G-medium. GLUT1, NGF, GDNF, VEGF, GCSF, and IGF2 levels were higher in S-medium than in G-medium, whereas FGF1, HIF, and MCP3 levels were not significantly different between media. In conclusion, we found that stem cell-conditioned medium had a neuroprotective effect against hypoxic injury, and that, of the various neurotrophic factors in S-medium, NGF, GDNF, and VEGF can contribute to neuroprotection. Copyright © 2017 Elsevier B.V. All rights reserved.

  18. Physiological temperature during brain slicing enhances the quality of acute slice preparations

    PubMed Central

    Huang, Shiwei; Uusisaari, Marylka Y.

    2013-01-01

    We demonstrate that brain dissection and slicing using solutions warmed to near-physiological temperature (~ +34°C), greatly enhance slice quality without affecting intrinsic electrophysiological properties of the neurons. Improved slice quality is seen not only when using young (<1 month), but also mature (>2.5 month) mice. This allows easy in vitro patch-clamp experimentation using adult deep cerebellar nuclear slices, which until now have been considered very difficult. As proof of the concept, we compare intrinsic properties of cerebellar nuclear neurons in juvenile (<1 month) and adult (up to 7 months) mice, and confirm that no significant developmental changes occur after the fourth postnatal week. The enhanced quality of brain slices from old animals facilitates experimentation on age-related disorders as well as optogenetic studies requiring long transfection periods. PMID:23630465

  19. Revealing 3D Ultrastructure and Morphology of Stem Cell Spheroids by Electron Microscopy.

    PubMed

    Jaros, Josef; Petrov, Michal; Tesarova, Marketa; Hampl, Ales

    2017-01-01

    Cell culture methods have been developed in efforts to produce biologically relevant systems for developmental and disease modeling, and appropriate analytical tools are essential. Knowledge of ultrastructural characteristics represents the basis to reveal in situ the cellular morphology, cell-cell interactions, organelle distribution, niches in which cells reside, and many more. The traditional method for 3D visualization of ultrastructural components, serial sectioning using transmission electron microscopy (TEM), is very labor-intensive due to contentious TEM slice preparation and subsequent image processing of the whole collection. In this chapter, we present serial block-face scanning electron microscopy, together with complex methodology for spheroid formation, contrasting of cellular compartments, image processing, and 3D visualization. The described technique is effective for detailed morphological analysis of stem cell spheroids, organoids, as well as organotypic cell cultures.

  20. Regulation of Axolotl (Ambystoma mexicanum) Limb Blastema Cell Proliferation by Nerves and BMP2 in Organotypic Slice Culture.

    PubMed

    Lehrberg, Jeffrey; Gardiner, David M

    2015-01-01

    We have modified and optimized the technique of organotypic slice culture in order to study the mechanisms regulating growth and pattern formation in regenerating axolotl limb blastemas. Blastema cells maintain many of the behaviors that are characteristic of blastemas in vivo when cultured as slices in vitro, including rates of proliferation that are comparable to what has been reported in vivo. Because the blastema slices can be cultured in basal medium without fetal bovine serum, it was possible to test the response of blastema cells to signaling molecules present in serum, as well as those produced by nerves. We also were able to investigate the response of blastema cells to experimentally regulated changes in BMP signaling. Blastema cells responded to all of these signals by increasing the rate of proliferation and the level of expression of the blastema marker gene, Prrx-1. The organotypic slice culture model provides the opportunity to identify and characterize the spatial and temporal co-regulation of pathways in order to induce and enhance a regenerative response.

  1. Regulation of Axolotl (Ambystoma mexicanum) Limb Blastema Cell Proliferation by Nerves and BMP2 in Organotypic Slice Culture

    PubMed Central

    Lehrberg, Jeffrey; Gardiner, David M.

    2015-01-01

    We have modified and optimized the technique of organotypic slice culture in order to study the mechanisms regulating growth and pattern formation in regenerating axolotl limb blastemas. Blastema cells maintain many of the behaviors that are characteristic of blastemas in vivo when cultured as slices in vitro, including rates of proliferation that are comparable to what has been reported in vivo. Because the blastema slices can be cultured in basal medium without fetal bovine serum, it was possible to test the response of blastema cells to signaling molecules present in serum, as well as those produced by nerves. We also were able to investigate the response of blastema cells to experimentally regulated changes in BMP signaling. Blastema cells responded to all of these signals by increasing the rate of proliferation and the level of expression of the blastema marker gene, Prrx-1. The organotypic slice culture model provides the opportunity to identify and characterize the spatial and temporal co-regulation of pathways in order to induce and enhance a regenerative response. PMID:25923915

  2. Mouse pancreas tissue slice culture facilitates long-term studies of exocrine and endocrine cell physiology in situ.

    PubMed

    Marciniak, Anja; Selck, Claudia; Friedrich, Betty; Speier, Stephan

    2013-01-01

    Studies on pancreatic cell physiology rely on the investigation of exocrine and endocrine cells in vitro. Particularly, in the case of the exocrine tissue these studies have suffered from a reduced functional viability of acinar cells in culture. As a result not only investigations on dispersed acinar cells and isolated acini were limited in their potential, but also prolonged studies on pancreatic exocrine and endocrine cells in an intact pancreatic tissue environment were unfeasible. To overcome these limitations, we aimed to establish a pancreas tissue slice culture platform to allow long-term studies on exocrine and endocrine cells in the intact pancreatic environment. Mouse pancreas tissue slice morphology was assessed to determine optimal long-term culture settings for intact pancreatic tissue. Utilizing optimized culture conditions, cell specificity and function of exocrine acinar cells and endocrine beta cells were characterized over a culture period of 7 days. We found pancreas tissue slices cultured under optimized conditions to have intact tissue specific morphology for the entire culture period. Amylase positive intact acini were present at all time points of culture and acinar cells displayed a typical strong cell polarity. Amylase release from pancreas tissue slices decreased during culture, but maintained the characteristic bell-shaped dose-response curve to increasing caerulein concentrations and a ca. 4-fold maximal over basal release. Additionally, endocrine beta cell viability and function was well preserved until the end of the observation period. Our results show that the tissue slice culture platform provides unprecedented maintenance of pancreatic tissue specific morphology and function over a culture period for at least 4 days and in part even up to 1 week. This analytical advancement now allows mid -to long-term studies on the cell biology of pancreatic disorder pathogenesis and therapy in an intact surrounding in situ.

  3. Mouse Pancreas Tissue Slice Culture Facilitates Long-Term Studies of Exocrine and Endocrine Cell Physiology in situ

    PubMed Central

    Marciniak, Anja; Selck, Claudia; Friedrich, Betty; Speier, Stephan

    2013-01-01

    Studies on pancreatic cell physiology rely on the investigation of exocrine and endocrine cells in vitro. Particularly, in the case of the exocrine tissue these studies have suffered from a reduced functional viability of acinar cells in culture. As a result not only investigations on dispersed acinar cells and isolated acini were limited in their potential, but also prolonged studies on pancreatic exocrine and endocrine cells in an intact pancreatic tissue environment were unfeasible. To overcome these limitations, we aimed to establish a pancreas tissue slice culture platform to allow long-term studies on exocrine and endocrine cells in the intact pancreatic environment. Mouse pancreas tissue slice morphology was assessed to determine optimal long-term culture settings for intact pancreatic tissue. Utilizing optimized culture conditions, cell specificity and function of exocrine acinar cells and endocrine beta cells were characterized over a culture period of 7 days. We found pancreas tissue slices cultured under optimized conditions to have intact tissue specific morphology for the entire culture period. Amylase positive intact acini were present at all time points of culture and acinar cells displayed a typical strong cell polarity. Amylase release from pancreas tissue slices decreased during culture, but maintained the characteristic bell-shaped dose-response curve to increasing caerulein concentrations and a ca. 4-fold maximal over basal release. Additionally, endocrine beta cell viability and function was well preserved until the end of the observation period. Our results show that the tissue slice culture platform provides unprecedented maintenance of pancreatic tissue specific morphology and function over a culture period for at least 4 days and in part even up to 1 week. This analytical advancement now allows mid -to long-term studies on the cell biology of pancreatic disorder pathogenesis and therapy in an intact surrounding in situ. PMID:24223842

  4. Organotypic slice cultures of human gastric and esophagogastric junction cancer.

    PubMed

    Koerfer, Justus; Kallendrusch, Sonja; Merz, Felicitas; Wittekind, Christian; Kubick, Christoph; Kassahun, Woubet T; Schumacher, Guido; Moebius, Christian; Gaßler, Nikolaus; Schopow, Nikolas; Geister, Daniela; Wiechmann, Volker; Weimann, Arved; Eckmann, Christian; Aigner, Achim; Bechmann, Ingo; Lordick, Florian

    2016-07-01

    Gastric and esophagogastric junction cancers are heterogeneous and aggressive tumors with an unpredictable response to cytotoxic treatment. New methods allowing for the analysis of drug resistance are needed. Here, we describe a novel technique by which human tumor specimens can be cultured ex vivo, preserving parts of the natural cancer microenvironment. Using a tissue chopper, fresh surgical tissue samples were cut in 400 μm slices and cultivated in 6-well plates for up to 6 days. The slices were processed for routine histopathology and immunohistochemistry. Cytokeratin stains (CK8, AE1/3) were applied for determining tumor cellularity, Ki-67 for proliferation, and cleaved caspase-3 staining for apoptosis. The slices were analyzed under naive conditions and following 2-4 days in vitro exposure to 5-FU and cisplatin. The slice culture technology allowed for a good preservation of tissue morphology and tumor cell integrity during the culture period. After chemotherapy exposure, a loss of tumor cellularity and an increase in apoptosis were observed. Drug sensitivity of the tumors could be assessed. Organotypic slice cultures of gastric and esophagogastric junction cancers were successfully established. Cytotoxic drug effects could be monitored. They may be used to examine mechanisms of drug resistance in human tissue and may provide a unique and powerful ex vivo platform for the prediction of treatment response. © 2016 The Authors. Cancer Medicine published by John Wiley & Sons Ltd.

  5. Ammonia-induced brain swelling and neurotoxicity in an organotypic slice model

    PubMed Central

    Back, Adam; Tupper, Kelsey Y.; Bai, Tao; Chiranand, Paulpoj; Goldenberg, Fernando D.; Frank, Jeffrey I.; Brorson, James R.

    2013-01-01

    Objectives Acute liver failure produces cerebral dysfunction and edema, mediated in part by elevated ammonia concentrations, often leading to coma and death. The pathophysiology of cerebral edema in acute liver failure is incompletely understood. In vitro models of the cerebral effects of acute liver failure have predominately consisted of dissociated astrocyte cultures or acute brain slices. We describe a stable long-term culture model incorporating both neural and glial elements in a three-dimensional tissue structure offering significant advantages to the study of astrocytic-neuronal interactions in the pathophysiology of cerebral edema and dysfunction in acute liver failure. Methods We utilized chronic organotypic slice cultures from mouse forebrain, applying ammonium acetate in iso-osmolar fashion for 72 hours. Imaging of slice thickness to assess for tissue swelling was accomplished in living slices with optical coherence tomography, and confocal microscopy of fluorescence immunochemical and histochemical staining served to assess astrocyte and neuronal numbers, morphology, and volume in the fixed brain slices. Results Ammonia exposure at 1–10 mM produced swelling of immunochemically-identified astrocytes, and at 10 mM resulted in macroscopic tissue swelling, with slice thickness increasing by about 30%. Astrocytes were unchanged in number. In contrast, 10 mM ammonia treatment severely disrupted neuronal morphology and reduced neuronal survival at 72 hours by one-half. Discussion Elevated ammonia produces astrocytic swelling, tissue swelling, and neuronal toxicity in cerebral tissues. Ammonia-treated organotypic brain slice cultures provide an in vitro model of cerebral effects of conditions relevant to acute liver failure, applicable to pathophysiological investigations. PMID:22196764

  6. Short-term field stimulation mimics synaptic maturation of hippocampal synapses

    PubMed Central

    Bagley, Elena E; Westbrook, Gary L

    2012-01-01

    Many aspects of synaptic transmission are modified during development, reflecting not only the consequence of developmental programmes of gene expression, but also the effects of ongoing neural activity. We investigated the role of synaptic activity in the maturation of Schaffer collateral (SC)–CA1 synapses using sustained low frequency field stimulation of acute brain slices. Between postnatal days 4–6 and 14–16, mouse SC–CA1 synapses in naïve slices showed a developmental decrease in the probability of transmitter release (Pr) and an increase in the contribution of GluN2A (NR2A) subunits to the NMDA receptor-mediated excitatory postsynaptic current (EPSC). Surprisingly, these developmental changes could be mimicked by short term (4 h) in vitro synaptic activity in slices taken from postnatal days (PND) 4–6 mice. However, different activity levels were required to alter release probability compared to the NMDA receptor subunit composition. Spontaneous synaptic activity was sufficient to alter the NMDA receptor subunit composition, but sustained low-frequency field stimulation of the brain slice (0.1 Hz, 4 h) was necessary to reduce release probability, as assessed 1 h following the cessation of stimulation. The protein synthesis inhibitor anisomycin blocked the effect of field stimulation on release probability. These results indicate that features of mature excitatory synapses can be rapidly induced in immature neurons. The activity dependence of the Pr and NMDA receptor subunit composition serves as a sensitive indicator of prior neural activity, and provides dual mechanisms for homeostatic control of excitatory synaptic efficacy. PMID:22351628

  7. Short-term field stimulation mimics synaptic maturation of hippocampal synapses.

    PubMed

    Bagley, Elena E; Westbrook, Gary L

    2012-04-01

    Many aspects of synaptic transmission are modified during development, reflecting not only the consequence of developmental programmes of gene expression, but also the effects of ongoing neural activity. We investigated the role of synaptic activity in the maturation of Schaffer collateral (SC)-CA1 synapses using sustained low frequency field stimulation of acute brain slices. Between postnatal days 4-6 and 14-16, mouse SC-CA1 synapses in naïve slices showed a developmental decrease in the probability of transmitter release (P(r)) and an increase in the contribution of GluN2A (NR2A) subunits to the NMDA receptor-mediated excitatory postsynaptic current (EPSC). Surprisingly, these developmental changes could be mimicked by short term (4 h) in vitro synaptic activity in slices taken from postnatal days (PND) 4-6 mice. However, different activity levels were required to alter release probability compared to the NMDA receptor subunit composition. Spontaneous synaptic activity was sufficient to alter the NMDA receptor subunit composition, but sustained low-frequency field stimulation of the brain slice (0.1 Hz, 4 h) was necessary to reduce release probability, as assessed 1 h following the cessation of stimulation. The protein synthesis inhibitor anisomycin blocked the effect of field stimulation on release probability. These results indicate that features of mature excitatory synapses can be rapidly induced in immature neurons. The activity dependence of the P(r) and NMDA receptor subunit composition serves as a sensitive indicator of prior neural activity, and provides dual mechanisms for homeostatic control of excitatory synaptic efficacy.

  8. Discovery of Hyperpolarized Molecular Imaging Biomarkers in a Novel Prostate Tissue Slice Culture Model

    DTIC Science & Technology

    2013-06-01

    start of tissue 2D culture , and 3 and 20 hours post culturing of the TSC’s in the 3D bioreactor (left) and corresponding representative tissue... culture tissue slice 13P spectrum obtained after 20 hours in the 5mm 3D MR compatible tissue culture bioreactor . 5 TSC’s with ≈ 90% viability...hours (≈98%) of being in 3D culture within the MR compatible bioreactor . • We demonstrated that the increased flux from pyruvate to lactate in the

  9. Inhibition Of Prostate Cancer Skeletal Metastases By Targeting Cathepsin K

    DTIC Science & Technology

    2010-02-01

    synthetic calcium phosphate thin films coated on to the culture vessels and on dentin slices in 96-wells plate. Soluble RANKL (50ng/ml) and MCSF (10ng/ml... dentin slices or synthetic calcium phosphate thin films are shown. Left panels without a frame: BD Biocate osteologic bone cell culture system, right...the 24-wells of BD osteologic bone cell culture system that consist of sub-micro synthetic calcium phosphate thin films coated onto the culture

  10. Electrophysiology of Hypothalamic Magnocellular Neurons In vitro: A Rhythmic Drive in Organotypic Cultures and Acute Slices.

    PubMed

    Israel, Jean-Marc; Oliet, Stéphane H; Ciofi, Philippe

    2016-01-01

    Hypothalamic neurohormones are released in a pulsatile manner. The mechanisms of this pulsatility remain poorly understood and several hypotheses are available, depending upon the neuroendocrine system considered. Among these systems, hypothalamo-neurohypophyseal magnocellular neurons have been early-considered models, as they typically display an electrical activity consisting of bursts of action potentials that is optimal for the release of boluses of the neurohormones oxytocin and vasopressin. The cellular mechanisms underlying this bursting behavior have been studied in vitro, using either acute slices of the adult hypothalamus, or organotypic cultures of neonatal hypothalamic tissue. We have recently proposed, from experiments in organotypic cultures, that specific central pattern generator networks, upstream of magnocellular neurons, determine their bursting activity. Here, we have tested whether a similar hypothesis can be derived from in vitro experiments in acute slices of the adult hypothalamus. To this aim we have screened our electrophysiological recordings of the magnocellular neurons, previously obtained from acute slices, with an analysis of autocorrelation of action potentials to detect a rhythmic drive as we recently did for organotypic cultures. This confirmed that the bursting behavior of magnocellular neurons is governed by central pattern generator networks whose rhythmic drive, and thus probably integrity, is however less satisfactorily preserved in the acute slices from adult brains.

  11. Inhibition of Prostate Cancer Skeletal Metastases by Targeting Cathepsin K

    DTIC Science & Technology

    2009-05-01

    micro synthetic calcium phosphate thin films coated onto the culture vessels. As a parallel study, a 96-well plate which contained dentin slice...bone resorption in vitro. (A) Representative images of resorption pits on dentin slices or synthetic calcium phosphate thin films are shown. Left...Osteologic Bone cell culture system (BD Bioscience) that consist of sub-micro synthetic calcium phosphate thin films coated on to the culture vessels and

  12. Effect of Noopept on Dynamics of Intracellular Calcium in Neurons of Cultured Rat Hippocampal Slices.

    PubMed

    Kolbaev, S N; Aleksandrova, O P; Sharonova, I N; Skrebitsky, V G

    2018-01-01

    A neuroprotective and nootropic drug Noopept increased the frequency of spontaneous calcium transients in neurons of CA1 radial layer in cultured rat hippocampal slices. In contrast, the drug exerted no significant effect on intracellular calcium concentration and its dynamics in neurons of hippocampal CA1 pyramidal layer.

  13. Essential role of the cAMP-cAMP response-element binding protein pathway in opiate-induced homeostatic adaptations of locus coeruleus neurons.

    PubMed

    Cao, Jun-Li; Vialou, Vincent F; Lobo, Mary Kay; Robison, Alfred J; Neve, Rachael L; Cooper, Donald C; Nestler, Eric J; Han, Ming-Hu

    2010-09-28

    Excessive inhibition of brain neurons in primary or slice cultures can induce homeostatic intrinsic plasticity, but the functional role and underlying molecular mechanisms of such plasticity are poorly understood. Here, we developed an ex vivo locus coeruleus (LC) slice culture system and successfully recapitulated the opiate-induced homeostatic adaptation in electrical activity of LC neurons seen in vivo. We investigated the mechanisms underlying this adaptation in LC slice cultures by use of viral-mediated gene transfer and genetic mutant mice. We found that short-term morphine treatment of slice cultures almost completely abolished the firing of LC neurons, whereas chronic morphine treatment increased LC neuronal excitability as revealed during withdrawal. This increased excitability was mediated by direct activation of opioid receptors and up-regulation of the cAMP pathway and accompanied by increased cAMP response-element binding protein (CREB) activity. Overexpression of a dominant negative CREB mutant blocked the increase in LC excitability induced by morphine- or cAMP-pathway activation. Knockdown of CREB in slice cultures from floxed CREB mice similarly decreased LC excitability. Furthermore, the ability of morphine or CREB overexpression to up-regulate LC firing was blocked by knockout of the CREB target adenylyl cyclase 8. Together, these findings provide direct evidence that prolonged exposure to morphine induces homeostatic plasticity intrinsic to LC neurons, involving up-regulation of the cAMP-CREB signaling pathway, which then enhances LC neuronal excitability.

  14. Ruminant organotypic brain-slice cultures as a model for the investigation of CNS listeriosis

    PubMed Central

    Guldimann, Claudia; Lejeune, Beatrice; Hofer, Sandra; Leib, Stephen L; Frey, Joachim; Zurbriggen, Andreas; Seuberlich, Torsten; Oevermann, Anna

    2012-01-01

    Central nervous system (CNS) infections in ruminant livestock, such as listeriosis, are of major concern for veterinary and public health. To date, no host-specific in vitro models for ruminant CNS infections are available. Here, we established and evaluated the suitability of organotypic brain-slices of ruminant origin as in vitro model to study mechanisms of Listeria monocytogenes CNS infection. Ruminants are frequently affected by fatal listeric rhombencephalitis that closely resembles the same condition occurring in humans. Better insight into host–pathogen interactions in ruminants is therefore of interest, not only from a veterinary but also from a public health perspective. Brains were obtained at the slaughterhouse, and hippocampal and cerebellar brain-slices were cultured up to 49 days. Viability as well as the composition of cell populations was assessed weekly. Viable neurons, astrocytes, microglia and oligodendrocytes were observed up to 49 days in vitro. Slice cultures were infected with L. monocytogenes, and infection kinetics were monitored. Infected brain cells were identified by double immunofluorescence, and results were compared to natural cases of listeric rhombencephalitis. Similar to the natural infection, infected brain-slices showed focal replication of L. monocytogenes and bacteria were predominantly observed in microglia, but also in astrocytes, and associated with axons. These results demonstrate that organotypic brain-slice cultures of bovine origin survive for extended periods and can be infected easily with L. monocytogenes. Therefore, they are a suitable model to study aspects of host–pathogen interaction in listeric encephalitis and potentially in other neuroinfectious diseases. PMID:22804762

  15. Modeling activity and target-dependent developmental cell death of mouse retinal ganglion cells ex vivo.

    PubMed

    Voyatzis, Sylvie; Muzerelle, Aude; Gaspar, Patricia; Nicol, Xavier

    2012-01-01

    Programmed cell death is widespread during the development of the central nervous system and serves multiple purposes including the establishment of neural connections. In the mouse retina a substantial reduction of retinal ganglion cells (RGCs) occurs during the first postnatal week, coinciding with the formation of retinotopic maps in the superior colliculus (SC). We previously established a retino-collicular culture preparation which recapitulates the progressive topographic ordering of RGC projections during early post-natal life. Here, we questioned whether this model could also be suitable to examine the mechanisms underlying developmental cell death of RGCs. Brn3a was used as a marker of the RGCs. A developmental decline in the number of Brn3a-immunolabelled neurons was found in the retinal explant with a timing that paralleled that observed in vivo. In contrast, the density of photoreceptors or of starburst amacrine cells increased, mimicking the evolution of these cell populations in vivo. Blockade of neural activity with tetrodotoxin increased the number of surviving Brn3a-labelled neurons in the retinal explant, as did the increase in target availability when one retinal explant was confronted with 2 or 4 collicular slices. Thus, this ex vivo model reproduces the developmental reduction of RGCs and recapitulates its regulation by neural activity and target availability. It therefore offers a simple way to analyze developmental cell death in this classic system. Using this model, we show that ephrin-A signaling does not participate to the regulation of the Brn3a population size in the retina, indicating that eprhin-A-mediated elimination of exuberant projections does not involve developmental cell death.

  16. Adaptation of Microplate-based Respirometry for Hippocampal Slices and Analysis of Respiratory Capacity

    PubMed Central

    Schuh, Rosemary A.; Clerc, Pascaline; Hwang, Hyehyun; Mehrabian, Zara; Bittman, Kevin; Chen, Hegang; Polster, Brian M.

    2011-01-01

    Multiple neurodegenerative disorders are associated with altered mitochondrial bioenergetics. Although mitochondrial O2 consumption is frequently measured in isolated mitochondria, isolated synaptic nerve terminals (synaptosomes), or cultured cells, the absence of mature brain circuitry is a remaining limitation. Here we describe the development of a method that adapts the Seahorse Extracellular Flux Analyzer (XF24) for the microplate-based measurement of hippocampal slice O2 consumption. As a first evaluation of the technique, we compared whole slice bioenergetics to previous measurements made with synaptosomes or cultured neurons. We found that mitochondrial respiratory capacity and O2 consumption coupled to ATP synthesis could be estimated in cultured or acute hippocampal slices with preserved neural architecture. Mouse organotypic hippocampal slices oxidizing glucose displayed mitochondrial O2 consumption that was well-coupled, as determined by the sensitivity to the ATP synthase inhibitor oligomycin. However stimulation of respiration by uncoupler was modest (<120% of basal respiration) compared to previous measurements in cells or synaptosomes, although enhanced slightly (to ~150% of basal respiration) by the acute addition of the mitochondrial complex I-linked substrate pyruvate. These findings suggest a high basal utilization of respiratory capacity in slices and a limitation of glucose-derived substrate for maximal respiration. The improved throughput of microplate-based hippocampal respirometry over traditional O2 electrode-based methods is conducive to neuroprotective drug screening. When coupled with cell type-specific pharmacology or genetic manipulations, the ability to efficiently measure O2 consumption from whole slices should advance our understanding of mitochondrial roles in physiology and neuropathology. PMID:21520220

  17. Low level postnatal methylmercury exposure in vivo alters developmental forms of short-term synaptic plasticity in the visual cortex of rat

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

    Dasari, Sameera; Yuan, Yukun, E-mail: yuanyuku@msu.ed

    2009-11-01

    Methylmercury (MeHg) has been previously shown to affect neurotransmitter release. Short-term synaptic plasticity (STP) is primarily related to changes in the probability of neurotransmitter release. To determine if MeHg affects STP development, we examined STP forms in the visual cortex of rat following in vivo MeHg exposure. Neonatal rats received 0 (0.9% NaCl), 0.75 or 1.5 mg/kg/day MeHg subcutaneously for 15 or 30 days beginning on postnatal day 5, after which visual cortical slices were prepared for field potential recordings. In slices prepared from rats treated with vehicle, field excitatory postsynaptic potentials (fEPSPs) evoked by paired-pulse stimulation at 20-200 msmore » inter-stimulus intervals showed a depression (PPD) of the second fEPSP (fEPSP2). PPD was also seen in slices prepared from rats after 15 day treatment with 0.75 or 1.5 mg/kg/day MeHg. However, longer duration treatment (30 days) with either dose of MeHg resulted in paired-pulse facilitation (PPF) of fEPSP2 in the majority of slices examined. PPF remained observable in slices prepared from animals in which MeHg exposure had been terminated for 30 days after completion of the initial 30 day MeHg treatment, whereas slices from control animals still showed PPD. MeHg did not cause any frequency- or region-preferential effect on STP. Manipulations of [Ca{sup 2+}]{sub e} or application of the GABA{sub A} receptor antagonist bicuculline could alter the strength and polarity of MeHg-induced changes in STP. Thus, these data suggest that low level postnatal MeHg exposure interferes with the developmental transformation of STP in the visual cortex, which is a long-lasting effect.« less

  18. In Situ Activation of Antigen-Specific CD8+ T Cells in the Presence of Antigen in Organotypic Brain Slices1

    PubMed Central

    Ling, Changying; Verbny, Yakov I.; Banks, Matthew I.; Sandor, Matyas; Fabry, Zsuzsanna

    2012-01-01

    The activation of Ag-specific T cells locally in the CNS could potentially contribute to the development of immune-mediated brain diseases. We addressed whether Ag-specific T cells could be stimulated in the CNS in the absence of peripheral lymphoid tissues by analyzing Ag-specific T cell responses in organotypic brain slice cultures. Organotypic brain slice cultures were established 1 h after intracerebral OVA Ag microinjection. We showed that when OVA-specific CD8+ T cells were added to Ag-containing brain slices, these cells became activated and migrated into the brain to the sites of their specific Ags. This activation of OVA-specific T cells was abrogated by the deletion of CD11c+ cells from the brain slices of the donor mice. These data suggest that brain-resident CD11c+ cells stimulate Ag-specific naive CD8+ T cells locally in the CNS and may contribute to immune responses in the brain. PMID:18523307

  19. Establishment and Characterization of a Tumor Stem Cell-Based Glioblastoma Invasion Model.

    PubMed

    Jensen, Stine Skov; Meyer, Morten; Petterson, Stine Asferg; Halle, Bo; Rosager, Ann Mari; Aaberg-Jessen, Charlotte; Thomassen, Mads; Burton, Mark; Kruse, Torben A; Kristensen, Bjarne Winther

    2016-01-01

    Glioblastoma is the most frequent and malignant brain tumor. Recurrence is inevitable and most likely connected to tumor invasion and presence of therapy resistant stem-like tumor cells. The aim was therefore to establish and characterize a three-dimensional in vivo-like in vitro model taking invasion and tumor stemness into account. Glioblastoma stem cell-like containing spheroid (GSS) cultures derived from three different patients were established and characterized. The spheroids were implanted in vitro into rat brain slice cultures grown in stem cell medium and in vivo into brains of immuno-compromised mice. Invasion was followed in the slice cultures by confocal time-lapse microscopy. Using immunohistochemistry, we compared tumor cell invasion as well as expression of proliferation and stem cell markers between the models. We observed a pronounced invasion into brain slice cultures both by confocal time-lapse microscopy and immunohistochemistry. This invasion closely resembled the invasion in vivo. The Ki-67 proliferation indexes in spheroids implanted into brain slices were lower than in free-floating spheroids. The expression of stem cell markers varied between free-floating spheroids, spheroids implanted into brain slices and tumors in vivo. The established invasion model kept in stem cell medium closely mimics tumor cell invasion into the brain in vivo preserving also to some extent the expression of stem cell markers. The model is feasible and robust and we suggest the model as an in vivo-like model with a great potential in glioma studies and drug discovery.

  20. A Device for Long-Term Perfusion, Imaging, and Electrical Interfacing of Brain Tissue In vitro

    PubMed Central

    Killian, Nathaniel J.; Vernekar, Varadraj N.; Potter, Steve M.; Vukasinovic, Jelena

    2016-01-01

    Distributed microelectrode array (MEA) recordings from consistent, viable, ≥500 μm thick tissue preparations over time periods from days to weeks may aid in studying a wide range of problems in neurobiology that require in vivo-like organotypic morphology. Existing tools for electrically interfacing with organotypic slices do not address necrosis that inevitably occurs within thick slices with limited diffusion of nutrients and gas, and limited removal of waste. We developed an integrated device that enables long-term maintenance of thick, functionally active, brain tissue models using interstitial perfusion and distributed recordings from thick sections of explanted tissue on a perforated multi-electrode array. This novel device allows for automated culturing, in situ imaging, and extracellular multi-electrode interfacing with brain slices, 3-D cell cultures, and potentially other tissue culture models. The device is economical, easy to assemble, and integrable with standard electrophysiology tools. We found that convective perfusion through the culture thickness provided a functional benefit to the preparations as firing rates were generally higher in perfused cultures compared to their respective unperfused controls. This work is a step toward the development of integrated tools for days-long experiments with more consistent, healthier, thicker, and functionally more active tissue cultures with built-in distributed electrophysiological recording and stimulation functionality. The results may be useful for the study of normal processes, pathological conditions, and drug screening strategies currently hindered by the limitations of acute (a few hours long) brain slice preparations. PMID:27065793

  1. Significant glial alterations in response to iron loading in a novel organotypic hippocampal slice culture model

    PubMed Central

    Healy, Sinead; McMahon, Jill; Owens, Peter; FitzGerald, Una

    2016-01-01

    Aberrant iron deposition in the brain is associated with neurodegenerative disorders including Multiple Sclerosis, Alzheimer’s disease and Parkinson’s disease. To study the collective response to iron loading, we have used hippocampal organotypic slices as a platform to develop a novel ex vivo model of iron accumulation. We demonstrated differential uptake and toxicity of iron after 12 h exposure to 10 μM ferrous ammonium sulphate, ferric citrate or ferrocene. Having established the supremacy of ferrocene in this model, the cultures were then loaded with 0.1–100 μM ferrocene for 12 h. One μM ferrocene exposure produced the maximal 1.6-fold increase in iron compared with vehicle. This was accompanied by a 1.4-fold increase in ferritin transcripts and mild toxicity. Using dual-immunohistochemistry, we detected ferritin in oligodendrocytes, microglia, but rarely in astrocytes and never in neurons in iron-loaded slice cultures. Moreover, iron loading led to a 15% loss of olig2-positive cells and a 16% increase in number and greater activation of microglia compared with vehicle. However, there was no appreciable effect of iron loading on astrocytes. In what we believe is a significant advance on traditional mono- or dual-cultures, our novel ex vivo slice-culture model allows characterization of the collective response of brain cells to iron-loading. PMID:27808258

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

    Hiser, C.; McIntosh, L.

    The rise in alternative respiratory capacity upon aging of potato (Solanum tuberosum) tuber slices is correlated with changes in mitochondrial membrane protein composition and a requirement for cytoplasmic protein synthesis. However, the lack of an antibody specific to the alternative oxidase has, until recently, prevented examination of the alternative oxidase protein(s) itself. We have employed a monoclonal antibody raised against the Sauromatum guttatum alternative oxidase to investigate developmental changes in the alternative pathway of aging potato slice mitochondria and to characterize the potato alternative oxidase by one- and two-dimensional gel electrophoresis. The relative levels of a 36 kilodalton protein parallelmore » the rise in alternative path capacity. A plausible interpretation is that this alternative oxidase protein is synthesized de novo during aging of potato slices.« less

  3. Preparation of organotypic brain slice cultures for the study of Alzheimer’s disease

    PubMed Central

    Croft, Cara L.; Noble, Wendy

    2018-01-01

    Alzheimer's disease, the most common cause of dementia, is a progressive neurodegenerative disorder characterised by amyloid-beta deposits in extracellular plaques, intracellular neurofibrillary tangles of aggregated tau, synaptic dysfunction and neuronal death. There are no cures for AD and current medications only alleviate some disease symptoms. Transgenic rodent models to study Alzheimer’s mimic features of human disease such as age-dependent accumulation of abnormal beta-amyloid and tau, synaptic dysfunction, cognitive deficits and neurodegeneration. These models have proven vital for improving our understanding of the molecular mechanisms underlying AD and for identifying promising therapeutic approaches. However, modelling neurodegenerative disease in animals commonly involves aging animals until they develop harmful phenotypes, often coupled with invasive procedures. In vivo studies are also resource, labour, time and cost intensive. We have developed a novel organotypic brain slice culture model to study Alzheimer’ disease which brings the potential of substantially reducing the number of rodents used in dementia research from an estimated 20,000 per year. We obtain 36 brain slices from each mouse pup, considerably reducing the numbers of animals required to investigate multiple stages of disease. This tractable model also allows the opportunity to modulate multiple pathways in tissues from a single animal. We believe that this model will most benefit dementia researchers in the academic and drug discovery sectors. We validated the slice culture model against aged mice, showing that the molecular phenotype closely mimics that displayed in vivo, albeit in an accelerated timescale. We showed beneficial outcomes following treatment of slices with agents previously shown to have therapeutic effects in vivo, and we also identified new mechanisms of action of other compounds. Thus, organotypic brain slice cultures from transgenic mouse models expressing Alzheimer’s disease-related genes may provide a valid and sensitive replacement for in vivo studies that do not involve behavioural analysis. PMID:29904599

  4. FGF-2 signal promotes proliferation of cerebellar progenitor cells and their oligodendrocytic differentiation at early postnatal stage

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

    Naruse, Masae; Shibasaki, Koji; Ishizaki, Yasuki, E-mail: yasukiishizaki@gunma-u.ac.jp

    The origins and developmental regulation of cerebellar oligodendrocytes are largely unknown, although some hypotheses of embryonic origins have been suggested. Neural stem cells exist in the white matter of postnatal cerebellum, but it is unclear whether these neural stem cells generate oligodendrocytes at postnatal stages. We previously showed that cerebellar progenitor cells, including neural stem cells, widely express CD44 at around postnatal day 3. In the present study, we showed that CD44-positive cells prepared from the postnatal day 3 cerebellum gave rise to neurospheres, while CD44-negative cells prepared from the same cerebellum did not. These neurospheres differentiated mainly into oligodendrocytesmore » and astrocytes, suggesting that CD44-positive neural stem/progenitor cells might generate oligodendrocytes in postnatal cerebellum. We cultured CD44-positive cells from the postnatal day 3 cerebellum in the presence of signaling molecules known as mitogens or inductive differentiation factors for oligodendrocyte progenitor cells. Of these, only FGF-2 promoted survival and proliferation of CD44-positive cells, and these cells differentiated into O4+ oligodendrocytes. Furthermore, we examined the effect of FGF-2 on cerebellar oligodendrocyte development ex vivo. FGF-2 enhanced proliferation of oligodendrocyte progenitor cells and increased the number of O4+ and CC1+ oligodendrocytes in slice cultures. These results suggest that CD44-positive cells might be a source of cerebellar oligodendrocytes and that FGF-2 plays important roles in their development at an early postnatal stage. - Highlights: • CD44 is expressed in cerebellar neural stem/progenitor cells at postnatal day 3 (P3). • FGF-2 promoted proliferation of CD44-positive progenitor cells from P3 cerebellum. • FGF-2 promoted oligodendrocytic differentiation of CD44-positive progenitor cells. • FGF-2 increased the number of oligodendrocytes in P3 cerebellar slice culture.« less

  5. Voltage-gated calcium channel autoimmune cerebellar degeneration

    PubMed Central

    McKasson, Marilyn; Clawson, Susan A.; Hill, Kenneth E.; Wood, Blair; Carlson, Noel; Bromberg, Mark; Greenlee, John E.

    2016-01-01

    Objectives: To describe response to treatment in a patient with autoantibodies against voltage-gated calcium channels (VGCCs) who presented with autoimmune cerebellar degeneration and subsequently developed Lambert-Eaton myasthenic syndrome (LEMS), and to study the effect of the patient's autoantibodies on Purkinje cells in rat cerebellar slice cultures. Methods: Case report and study of rat cerebellar slice cultures incubated with patient VGCC autoantibodies. Results: A 53-year-old man developed progressive incoordination with ataxic speech. Laboratory evaluation revealed VGCC autoantibodies without other antineuronal autoantibodies. Whole-body PET scans 6 and 12 months after presentation detected no malignancy. The patient improved significantly with IV immunoglobulin G (IgG), prednisone, and mycophenolate mofetil, but worsened after IV IgG was halted secondary to aseptic meningitis. He subsequently developed weakness with electrodiagnostic evidence of LEMS. The patient's IgG bound to Purkinje cells in rat cerebellar slice cultures, followed by neuronal death. Reactivity of the patient's autoantibodies with VGCCs was confirmed by blocking studies with defined VGCC antibodies. Conclusions: Autoimmune cerebellar degeneration associated with VGCC autoantibodies may precede onset of LEMS and may improve with immunosuppressive treatment. Binding of anti-VGCC antibodies to Purkinje cells in cerebellar slice cultures may be followed by cell death. Patients with anti-VGCC autoantibodies may be at risk of irreversible neurologic injury over time, and treatment should be initiated early. PMID:27088118

  6. PROPYLTHIOURACIL (PTU)-INDUCED HYPOTHYROIDISM: EFFECTS ON SYNAPTIC TRANSMISSION AND LONG TERM POTENTIATION IN HIPPOCAMPAL SLICES.

    EPA Science Inventory

    Concern has been raised over endocrine effects of some classes of environmental chemicals. Severe hypothyroidism during critical periods of brain developmental leads to alterations in hippocampal structure, learning deficits, yet neurophysiological properties of the hippocampus...

  7. Marked synergism between mutant SOD1 and glutamate transport inhibition in the induction of motor neuronal degeneration in spinal cord slice cultures.

    PubMed

    Yin, Hong Z; Weiss, John H

    2012-04-11

    Loss of astrocytic glutamate transport capacity in ALS spinal cord supports an excitotoxic contribution to motor neuron (MN) damage in the disease, and dominant gain of function mutations in Cu/Zn superoxide dismutase (SOD1) cause certain familial forms of ALS. We have used organotypic slice cultures from wild type and G93A SOD1 mutant rat spinal cords to examine interactions between excitotoxicity and the presence of mutant SOD1 in the induction of MN degeneration. Slice cultures were prepared from 1 week old pups, and after an additional week in vitro, some were exposed to either a low level (30 μM) of the glutamate uptake inhibitor, trans-pyrrolidine-2,4-dicarboxylic acid (PDC) for 3 weeks, or a higher level (50 μM) for 48 h, followed by histochemical labeling to assess MN injury. In wild type animals these exposures caused relatively little MN degeneration. Similarly, little MN degeneration was seen in slices from SOD1 mutant animals that were not exposed to PDC. However, addition of PDC to SOD1 mutant slices resulted in substantial MN injury, which was markedly attenuated by a Ca2+ permeable AMPA-type (Ca-AMPA) glutamate channel blocker, or by a nitric oxide synthase antagonist. These observations illustrate the utility of the organotypic culture model for the investigation of intracellular interactions underlying MN degeneration in ALS, and support the hypothesis that activation of Ca-AMPA channels on MNs provides a metabolic burden that synergizes with deleterious effects of mutant SOD1 in the induction of MN injury. Copyright © 2012 Elsevier B.V. All rights reserved.

  8. A co-culture system with an organotypic lung slice and an immortal alveolar macrophage cell line to quantify silica-induced inflammation.

    PubMed

    Hofmann, Falk; Bläsche, Robert; Kasper, Michael; Barth, Kathrin

    2015-01-01

    There is growing evidence that amorphous silica nanoparticles cause toxic effects on lung cells in vivo as well as in vitro and induce inflammatory processes. The phagocytosis of silica by alveolar macrophages potentiates these effects. To understand the underlying molecular mechanisms of silica toxicity, we applied a co-culture system including the immortal alveolar epithelial mouse cell line E10 and the macrophage cell line AMJ2-C11. In parallel we exposed precision-cut lung slices (lacking any blood cells as well as residual alveolar macrophages) of wild type and P2rx7-/- mice with or without AMJ2-C11 cells to silica nanoparticles. Exposure of E10 cells as well as slices of wild type mice resulted in an increase of typical alveolar epithelial type 1 cell proteins like T1α, caveolin-1 and -2 and PKC-β1, whereas the co-culture with AMJ2-C11 showed mostly a slightly lesser increase of these proteins. In P2rx7-/- mice most of these proteins were slightly decreased. ELISA analysis of the supernatant of wild type and P2rx7-/- mice precision-cut lung slices showed decreased amounts of IL-6 and TNF-α when incubated with nano-silica. Our findings indicate that alveolar macrophages influence the early inflammation of the lung and also that cell damaging reagents e.g. silica have a smaller impact on P2rx7-/- mice than on wild type mice. The co-culture system with an organotypic lung slice is a useful tool to study the role of alveolar macrophages during lung injury at the organoid level.

  9. Protocols and characterization data for 2D, 3D, and slice-based tumor models from the PREDECT project.

    PubMed

    de Hoogt, Ronald; Estrada, Marta F; Vidic, Suzana; Davies, Emma J; Osswald, Annika; Barbier, Michael; Santo, Vítor E; Gjerde, Kjersti; van Zoggel, Hanneke J A A; Blom, Sami; Dong, Meng; Närhi, Katja; Boghaert, Erwin; Brito, Catarina; Chong, Yolanda; Sommergruber, Wolfgang; van der Kuip, Heiko; van Weerden, Wytske M; Verschuren, Emmy W; Hickman, John; Graeser, Ralph

    2017-11-21

    Two-dimensional (2D) culture of cancer cells in vitro does not recapitulate the three-dimensional (3D) architecture, heterogeneity and complexity of human tumors. More representative models are required that better reflect key aspects of tumor biology. These are essential studies of cancer biology and immunology as well as for target validation and drug discovery. The Innovative Medicines Initiative (IMI) consortium PREDECT (www.predect.eu) characterized in vitro models of three solid tumor types with the goal to capture elements of tumor complexity and heterogeneity. 2D culture and 3D mono- and stromal co-cultures of increasing complexity, and precision-cut tumor slice models were established. Robust protocols for the generation of these platforms are described. Tissue microarrays were prepared from all the models, permitting immunohistochemical analysis of individual cells, capturing heterogeneity. 3D cultures were also characterized using image analysis. Detailed step-by-step protocols, exemplary datasets from the 2D, 3D, and slice models, and refined analytical methods were established and are presented.

  10. Protocols and characterization data for 2D, 3D, and slice-based tumor models from the PREDECT project

    PubMed Central

    de Hoogt, Ronald; Estrada, Marta F.; Vidic, Suzana; Davies, Emma J.; Osswald, Annika; Barbier, Michael; Santo, Vítor E.; Gjerde, Kjersti; van Zoggel, Hanneke J. A. A.; Blom, Sami; Dong, Meng; Närhi, Katja; Boghaert, Erwin; Brito, Catarina; Chong, Yolanda; Sommergruber, Wolfgang; van der Kuip, Heiko; van Weerden, Wytske M.; Verschuren, Emmy W.; Hickman, John; Graeser, Ralph

    2017-01-01

    Two-dimensional (2D) culture of cancer cells in vitro does not recapitulate the three-dimensional (3D) architecture, heterogeneity and complexity of human tumors. More representative models are required that better reflect key aspects of tumor biology. These are essential studies of cancer biology and immunology as well as for target validation and drug discovery. The Innovative Medicines Initiative (IMI) consortium PREDECT (www.predect.eu) characterized in vitro models of three solid tumor types with the goal to capture elements of tumor complexity and heterogeneity. 2D culture and 3D mono- and stromal co-cultures of increasing complexity, and precision-cut tumor slice models were established. Robust protocols for the generation of these platforms are described. Tissue microarrays were prepared from all the models, permitting immunohistochemical analysis of individual cells, capturing heterogeneity. 3D cultures were also characterized using image analysis. Detailed step-by-step protocols, exemplary datasets from the 2D, 3D, and slice models, and refined analytical methods were established and are presented. PMID:29160867

  11. Kynurenine 3-mono-oxygenase inhibitors attenuate post-ischemic neuronal death in organotypic hippocampal slice cultures.

    PubMed

    Carpenedo, Raffaella; Meli, Elena; Peruginelli, Fiamma; Pellegrini-Giampietro, Domenico E; Moroni, Flavio

    2002-09-01

    Kynurenine 3-mono-oxygenase (KMO) inhibitors reduce 3-hydroxykynurenine (3-HK) and quinolinic acid (QUIN) neosynthesis and facilitate kynurenine metabolism towards kynurenic acid (KYNA) formation. They also reduce tissue damage in models of focal or transient global cerebral ischemia in vivo. We used organotypic hippocampal slice cultures exposed to oxygen and glucose deprivation (OGD) to investigate KMO mechanism(s) of neuroprotective activity. Exposure of the slices to 30 min of OGD caused CA1 pyramidal cell death and significantly decreased the amount of KYNA released in the incubation medium. The KMO inhibitors (m-nitrobenzoyl)-alanine (30-100 micro m) or 3,4-dimethoxy-[-N-4-(nitrophenyl)thiazol-2yl]-benzenesulfonamide (1-10 micro m) reduced post-ischemic neuronal death and increased KYNA concentrations in slice incubation media. The maximal concentration of KYNA detected in the incubation media of slices treated with KMO inhibitors was approximately 50 nm and was too low to efficiently interact with alpha7 nicotinic acetylcholine receptors or with the glycineb site of N-methyl-d-aspartate (NMDA) receptors. On the other hand, the addition of either 3-HK or QUIN (1-10 micro m) to OGD-exposed hippocampal slices prevented the neuroprotective activity of KMO inhibitors. Our results suggest that KMO inhibitors reduce the neuronal death found in the CA1 region of organotypic hippocampal slices exposed to 30 min of OGD by decreasing the local synthesis of 3-HK and QUIN.

  12. Precision cut lung slices as an efficient tool for in vitro lung physio-pharmacotoxicology studies.

    PubMed

    Morin, Jean-Paul; Baste, Jean-Marc; Gay, Arnaud; Crochemore, Clément; Corbière, Cécile; Monteil, Christelle

    2013-01-01

    1.We review the specific approaches for lung tissue slices preparation and incubation systems and the research application fields in which lung slices proved to be a very efficient alternative to animal experimentation for biomechanical, physiological, pharmacological and toxicological approaches. 2.Focus is made on air-liquid interface dynamic organ culture systems that allow direct tissue exposure to complex aerosol and that best mimic in vivo lung tissue physiology. 3.A compilation of research applications in the fields of vascular and airway reactivity, mucociliary transport, polyamine transport, xenobiotic biotransformation, chemicals toxicology and complex aerosols supports the concept that precision cut lung slices are a very efficient tool maintaining highly differentiated functions similar to in vivo lung organ when kept under dynamic organ culture. They also have been successfully used for lung gene transfer efficiency assessment, for lung viral infection efficiency assessment, for studies of tissue preservation media and tissue post-conditioning to optimize lung tissue viability before grafting. 4.Taken all together, the reviewed studies point to a great interest for precision cut lung slices as an efficient and valuable alternative to in vivo lung organ experimentation.

  13. Trafficking of astrocytic vesicles in hippocampal slices

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

    Potokar, Maja; Kreft, Marko; Celica Biomedical Center, Technology Park 24, 1000 Ljubljana

    2009-12-25

    The increasingly appreciated role of astrocytes in neurophysiology dictates a thorough understanding of the mechanisms underlying the communication between astrocytes and neurons. In particular, the uptake and release of signaling substances into/from astrocytes is considered as crucial. The release of different gliotransmitters involves regulated exocytosis, consisting of the fusion between the vesicle and the plasma membranes. After fusion with the plasma membrane vesicles may be retrieved into the cytoplasm and may continue to recycle. To study the mobility implicated in the retrieval of secretory vesicles, these structures have been previously efficiently and specifically labeled in cultured astrocytes, by exposing livemore » cells to primary and secondary antibodies. Since the vesicle labeling and the vesicle mobility properties may be an artifact of cell culture conditions, we here asked whether the retrieving exocytotic vesicles can be labeled in brain tissue slices and whether their mobility differs to that observed in cell cultures. We labeled astrocytic vesicles and recorded their mobility with two-photon microscopy in hippocampal slices from transgenic mice with fluorescently tagged astrocytes (GFP mice) and in wild-type mice with astrocytes labeled by Fluo4 fluorescence indicator. Glutamatergic vesicles and peptidergic granules were labeled by the anti-vesicular glutamate transporter 1 (vGlut1) and anti-atrial natriuretic peptide (ANP) antibodies, respectively. We report that the vesicle mobility parameters (velocity, maximal displacement and track length) recorded in astrocytes from tissue slices are similar to those reported previously in cultured astrocytes.« less

  14. Long Term Synaptic Plasticity and Learning in Neuronal Networks

    DTIC Science & Technology

    1989-01-14

    Videomicroscopy and synaptic physiology of cultured hippocampal slices. Soc, Neurosci. Abstr. 14:246, 1988. Griffith, W.H., Brown, T.H. and Johnston, D...Chapman, P.F., Chang, V., and Brown, T.H. . Videomicroscopy of acute brain slices from hippocampus and amygdala. Brain Res. Bull, 21: 373-383, 1988

  15. Through Two Lenses: A Cultural-Developmental Approach to Moral Psychology

    ERIC Educational Resources Information Center

    Jensen, Lene Arnett

    2008-01-01

    This paper proposes a cultural-developmental approach to moral psychology. The approach builds on and synthesizes findings from different research traditions, including the cognitive-developmental, domain, two orientations, three ethics, and moral identity traditions. The paper introduces a conception termed a "cultural-developmental template."…

  16. N-methyl-D-aspartate neurotoxicity in hippocampal slices: protection by aniracetam.

    PubMed

    Pizzi, M; Consolandi, O; Memo, M; Spano, P

    1995-03-14

    Aniracetam, a drug known to elicit cognition enhancing properties in both animals and humans, was found to counteract the neurotoxicity induced by excitatory amino acids in primary cultures of cerebellar neurons. We report here that aniracetam prevents the neurotoxic effect induced by N-methyl-D-aspartate (NMDA) in rat hippocampal slices. Time-course experiments showed that the aniracetam-induced neuroprotection does not require preincubation of the slices with the drug. Maximal effective concentration of aniracetam was 10 microM. Since the NMDA-mediated cell death in hippocampal slices is considered a valuable experimental model of ischemia, these results suggest a possible novel therapeutic application for aniracetam.

  17. Gender-Fair and Gender-Congruent Practices for Young Children's Naturalist Intelligence: From the Perspective of Developmentally and Culturally Appropriate Practice (DCAP).

    ERIC Educational Resources Information Center

    Hyun, Eunsook

    Developmentally and Culturally Appropriate Practice (DCAP) is a culturally congruent and critical pedagogy that serves as a framework for early childhood education for all individuals. This paper examines young children's gender differences in learning and their gender-oriented culture and promotes developmentally and culturally appropriate…

  18. Early increase and late decrease of purkinje cell dendritic spine density in prion-infected organotypic mouse cerebellar cultures.

    PubMed

    Campeau, Jody L; Wu, Gengshu; Bell, John R; Rasmussen, Jay; Sim, Valerie L

    2013-01-01

    Prion diseases are infectious neurodegenerative diseases associated with the accumulation of protease-resistant prion protein, neuronal loss, spongiform change and astrogliosis. In the mouse model, the loss of dendritic spines is one of the earliest pathological changes observed in vivo, occurring 4-5 weeks after the first detection of protease-resistant prion protein in the brain. While there are cell culture models of prion infection, most do not recapitulate the neuropathology seen in vivo. Only the recently developed prion organotypic slice culture assay has been reported to undergo neuronal loss and the development of some aspects of prion pathology, namely small vacuolar degeneration and tubulovesicular bodies. Given the rapid replication of prions in this system, with protease-resistant prion protein detectable by 21 days, we investigated whether the dendritic spine loss and altered dendritic morphology seen in prion disease might also develop within the lifetime of this culture system. Indeed, six weeks after first detection of protease-resistant prion protein in tga20 mouse cerebellar slice cultures infected with RML prion strain, we found a statistically significant loss of Purkinje cell dendritic spines and altered dendritic morphology in infected cultures, analogous to that seen in vivo. In addition, we found a transient but statistically significant increase in Purkinje cell dendritic spine density during infection, at the time when protease-resistant prion protein was first detectable in culture. Our findings support the use of this slice culture system as one which recapitulates prion disease pathology and one which may facilitate study of the earliest stages of prion disease pathogenesis.

  19. Blast waves from detonated military explosive reduce GluR1 and synaptophysin levels in hippocampal slice cultures✩

    PubMed Central

    Smith, Marquitta; Piehler, Thuvan; Benjamin, Richard; Farizatto, Karen L.; Pait, Morgan C.; Almeida, Michael F.; Ghukasyan, Vladimir V.; Bahr, Ben A.

    2017-01-01

    Explosives create shockwaves that cause blast-induced neurotrauma, one of the most common types of traumatic brain injury (TBI) linked to military service. Blast-induced TBIs are often associated with reduced cognitive and behavioral functions due to a variety of factors. To study the direct effects of military explosive blasts on brain tissue, we removed systemic factors by utilizing rat hippocampal slice cultures. The long-term slice cultures were briefly sealed air-tight in serum-free medium, lowered into a 37 °C water-filled tank, and small 1.7-gram assemblies of cyclotrimethylene trinitramine (RDX) were detonated 15 cm outside the tank, creating a distinct shockwave recorded at the culture plate position. Compared to control mock-treated groups of slices that received equal submerge time, 1–3 blast impacts caused a dose-dependent reduction in the AMPA receptor subunit GluR1. While only a small reduction was found in hippocampal slices exposed to a single RDX blast and harvested 1–2 days later, slices that received two consecutive RDX blasts 4 min apart exhibited a 26–40% reduction in GluR1, and the receptor subunit was further reduced by 64–72% after three consecutive blasts. Such loss correlated with increased levels of HDAC2, a histone deacetylase implicated in stress-induced reduction of glutamatergic transmission. No evidence of synaptic marker recovery was found at 72 h post-blast. The presynaptic marker synaptophysin was found to have similar susceptibility as GluR1 to the multiple explosive detonations. In contrast to the synaptic protein reductions, actin levels were unchanged, spectrin breakdown was not detected, and Fluoro-Jade B staining found no indication of degenerating neurons in slices exposed to three RDX blasts, suggesting that small, sub-lethal explosives are capable of producing selective alterations to synaptic integrity. Together, these results indicate that blast waves from military explosive cause signs of synaptic compromise without producing severe neurodegeneration, perhaps explaining the cognitive and behavioral changes in those blast-induced TBI sufferers that have no detectable neuropathology. PMID:27720798

  20. Tissue culture on a chip: Developmental biology applications of self-organized capillary networks in microfluidic devices.

    PubMed

    Miura, Takashi; Yokokawa, Ryuji

    2016-08-01

    Organ culture systems are used to elucidate the mechanisms of pattern formation in developmental biology. Various organ culture techniques have been used, but the lack of microcirculation in such cultures impedes the long-term maintenance of larger tissues. Recent advances in microfluidic devices now enable us to utilize self-organized perfusable capillary networks in organ cultures. In this review, we will overview past approaches to organ culture and current technical advances in microfluidic devices, and discuss possible applications of microfluidics towards the study of developmental biology. © 2016 Japanese Society of Developmental Biologists.

  1. Effect of embryo density on in vitro developmental characteristics of bovine preimplantative embryos with respect to micro and macroenvironments.

    PubMed

    Hoelker, M; Rings, F; Lund, Q; Phatsara, C; Schellander, K; Tesfaye, D

    2010-10-01

    To overcome developmental problems as a consequence of single embryo culture, the Well of the Well (WOW) culture system has been developed. In this study, we aimed to examine the effect of embryo densities with respect to both microenvironment and macroenvironment on developmental rates and embryo quality to get a deeper insight into developmentally important mechanisms. WOW diameter and depth significantly affected developmental rates (p < 0.05). WOWs with diameter of 500 μm reached significantly higher blastocyst rates (32.5 vs 21.1% vs 20.3%) compared to embryos cultured in WOWs of 300 μm diameter or plain cultured controls. Embryos cultured in WOWs with 700 μm depth reached significant higher developmental rates compared with embryos cultured in WOWs of 300 μm depth and control embryos (30.6 vs 22.6% vs 20.3%). Correlation of the embryo per WOW volume with developmental rates was higher (r(2) = 0.92, p = 0.0004) than correlation of WOW diameter or WOW depth with developmental rates. However, the embryo per WOW volume did not affect differential cell counts. An embryo per culture dish volume of 1 : 30 μl was identified to be optimal when the embryo per WOW volume was 1 : 0.27 μl increasing developmental rates up to the level of mass embryo production. Giving the opportunity to track each embryo over the complete culture period while keeping high developmental rates with normal mitotic dynamics, the results of this work will provide benefit for the single culture of embryos in human assisted reproduction, mammalian embryos with high economic interest as well as for scientific purpose. © 2009 Blackwell Verlag GmbH.

  2. Long-term, repeated dose in vitro neurotoxicity of the glutamate receptor antagonist L-AP3, demonstrated in rat hippocampal slice cultures by using continuous propidium iodide incubation.

    PubMed

    Kristensen, Bjarne W; Blaabjerg, Morten; Noraberg, Jens; Zimmer, Jens

    2007-05-01

    Most in vitro models are only used to assess short-term effects of test compounds. However, as demonstrated here, hippocampal slice cultures can be used for long-term studies. The test compound used was the metabotropic glutamate receptor antagonist, L(+)-2-amino-3-phosphonopropionic acid (L-AP3), which is known to be toxic in vivo after subchronic, but not acute, administration. Degenerative effects were monitored by measuring the cellular uptake of propidium iodide (PI; continuously present in the medium) and lactate dehydrogenase (LDH) leakage, and by using a panel of histological stains. Hippocampal slices, derived from 2-3 day old rats and grown for 3 weeks, were subsequently exposed for the next 3 weeks to 0, 10 or 100microM L-AP3, with PI (2microM) in the culture medium. Exposure to 100microM L-AP3 induced severe toxicity after 4-6 days, shown by massive PI uptake, LDH leakage, changes in MAP2 and GFAP immunostaining, and in Nissl and Timm staining. In contrast, 10microM L-AP3 did not induce detectable neuronal degeneration. Treatment with the NMDA receptor antagonist, MK-801, or the AMPA/KA receptor antagonist NBQX, together with 100microM L-AP3, reduced neurodegeneration down to close to control values. It is concluded that continuous incubation of hippocampal slice cultures with PI is technically feasible for use in studies of inducible neuronal degeneration over time.

  3. Follicle dynamics: visualization and analysis of follicle growth and maturation using murine ovarian tissue culture.

    PubMed

    Murase, Tomohiko; Iwase, Akira; Komatsu, Kouji; Bayasula; Nakamura, Tomoko; Osuka, Satoko; Takikawa, Sachiko; Goto, Maki; Kotani, Tomomi; Kikkawa, Fumitaka

    2018-02-01

    To visualize and analyze follicle development in ovarian tissue culture using physiological concentrations of follicle-stimulating hormone (FSH) and luteinizing hormone (LH) in order to establish an ovarian tissue culture system that enables efficient in vitro growth of follicles. Ovarian tissues from 4-week-old female ICR mice were sliced and cultured. Images of ovarian tissues in culture were obtained at 24-h or 30-min intervals by using a microscope. The area of each follicle observed in the ovarian tissue slices was tracked and analyzed in association with oocyte maturation. We were able to track the development of each follicle using this culture system. Follicle growth was associated with oocyte maturation. Meiotically matured oocytes (MII) were obtained from 33% of all follicles investigated. Approximately, a quarter of follicles (24%) did not grow and resulted in atresia. Follicle dynamics were successfully visualized and analyzed in murine ovarian tissue culture. We were able to obtain mature oocytes from the fully grown follicles in vitro. This culture system would be helpful for efficient in vitro culturing of ovarian tissues.

  4. A brain slice bath for physiology and compound microscopy, with dual-sided perifusion.

    PubMed

    Heyward, P M

    2010-12-01

    Contemporary in vitro brain slice studies can employ compound microscopes to identify individual neurons or their processes for physiological recording or imaging. This requires that the bath used to maintain the tissue fits within the working distances of a water-dipping objective and microscope condenser. A common means of achieving this is to maintain thin tissue slices on the glass floor of a recording bath, exposing only one surface of the tissue to oxygenated bathing medium. Emerging evidence suggests that physiology can be compromised by this approach. Flowing medium past both sides of submerged brain slices is optimal, but recording baths utilizing this principle are not readily available for use on compound microscopes. This paper describes a tissue bath designed specifically for microscopy and physiological recording, in which temperature-controlled medium flows past both sides of the slices. A particular feature of this design is the use of concentric mesh rings to support and transport the live tissue without mechanical disturbance. The design is also easily adapted for use with thin acute slices, cultured slices, and acutely dispersed or cultured cells maintained either on cover slips or placed directly on the floor of the bath. The low profile of the bath provides a low angle of approach for electrodes, and allows use of standard condensers, nosepieces and water-dipping objective lenses. If visualization of individual neurons is not required, the bath can be mounted on a simple stand and used with a dissecting microscope. Heating is integral to the bath, and any temperature controller capable of driving a resistive load can be used. The bath is robust, readily constructed and requires minimal maintenance. Full construction and operation details are given. © 2010 The Author Journal of Microscopy © 2010 The Royal Microscopical Society.

  5. Perfusion cell culture decreases process and product heterogeneity in a head-to-head comparison with fed-batch.

    PubMed

    Walther, Jason; Lu, Jiuyi; Hollenbach, Myles; Yu, Marcella; Hwang, Chris; McLarty, Jean; Brower, Kevin

    2018-05-30

    In this study, we compared the impacts of fed-batch and perfusion platforms on process and product attributes for IgG1- and IgG4-producing cell lines. A "plug-and-play" approach was applied to both platforms at bench scale, using commercially available basal and feed media, a standard feed strategy for fed-batch, and ATF filtration for perfusion. Product concentration in fed-batch was 2.5 times greater than perfusion, while average productivity in perfusion was 7.5 times greater than fed-batch. PCA revealed more variability in the cell environment and metabolism during the fed-batch run. LDH measurements showed that exposure of product to cell lysate was 7-10 times greater in fed-batch. Product analysis shows larger abundances of neutral species in perfusion, likely due to decreased bioreactor residence times and extracellular exposure. The IgG1 perfusion product also had higher purity and lower half-antibody. Glycosylation was similar across both culture modes. The first perfusion harvest slice for both product types showed different glycosylation than subsequent harvests, suggesting that product quality lags behind metabolism. In conclusion, process and product data indicate that intra-lot heterogeneity is decreased in perfusion cultures. Additional data and discussion is required to understand the developmental, clinical and commercial implications, and in what situations increased uniformity would be beneficial. This article is protected by copyright. All rights reserved.

  6. Action potentials reliably invade axonal arbors of rat neocortical neurons

    PubMed Central

    Cox, Charles L.; Denk, Winfried; Tank, David W.; Svoboda, Karel

    2000-01-01

    Neocortical pyramidal neurons have extensive axonal arborizations that make thousands of synapses. Action potentials can invade these arbors and cause calcium influx that is required for neurotransmitter release and excitation of postsynaptic targets. Thus, the regulation of action potential invasion in axonal branches might shape the spread of excitation in cortical neural networks. To measure the reliability and extent of action potential invasion into axonal arbors, we have used two-photon excitation laser scanning microscopy to directly image action-potential-mediated calcium influx in single varicosities of layer 2/3 pyramidal neurons in acute brain slices. Our data show that single action potentials or bursts of action potentials reliably invade axonal arbors over a range of developmental ages (postnatal 10–24 days) and temperatures (24°C-30°C). Hyperpolarizing current steps preceding action potential initiation, protocols that had previously been observed to produce failures of action potential propagation in cultured preparations, were ineffective in modulating the spread of action potentials in acute slices. Our data show that action potentials reliably invade the axonal arbors of neocortical pyramidal neurons. Failures in synaptic transmission must therefore originate downstream of action potential invasion. We also explored the function of modulators that inhibit presynaptic calcium influx. Consistent with previous studies, we find that adenosine reduces action-potential-mediated calcium influx in presynaptic terminals. This reduction was observed in all terminals tested, suggesting that some modulatory systems are expressed homogeneously in most terminals of the same neuron. PMID:10931955

  7. Effects of endogenous pyrogen and prostaglandin E2 on hypothalamic neurons in rat brain slices.

    PubMed

    Watanabe, T; Morimoto, A; Murakami, N

    1987-06-01

    We investigated the effects of endogenous pyrogen and prostaglandin E2 (PGE2) on the preoptic and anterior hypothalamic (POAH) neurons using brain slice preparations from the rat. Partially purified endogenous pyrogen did not change the activities of most of the neurons in the POAH region when applied locally through a micropipette attached to the recording electrode in proximity to the neurons. This indicates that partially purified endogenous pyrogen does not act directly on the neuronal activity in the POAH region. The partially purified endogenous pyrogen, applied into a culture chamber containing a brain slice, facilitated the activities in 24% of the total neurons tested, regardless of the thermal specificity of the neurons. Moreover, PGE2 added to the culture chamber facilitated 48% of the warm-responsive, 33% of the cold-responsive, and 29% of the thermally insensitive neurons. The direction of change in neuronal activity induced by partially purified endogenous pyrogen appears to be almost the same as that induced by PGE2 when these substances were applied by perfusion to the same neuron in the culture chamber. These results suggest that partially purified pyrogen applied to the perfusate of the culture chamber stimulates some constituents of brain tissue to synthesize and release prostaglandin, which in turn affects the neuronal activity of the POAH region.

  8. Essential Role of Culture in Developmental Psychology

    ERIC Educational Resources Information Center

    Miller, Joan G.

    2005-01-01

    This chapter argues for the essential role of culture in forming the basic constructs and theories of developmental psychology. The case is made for the need to overcome the cultural insularity of core developmental concepts and methods in order to create a psychology that is more truly universal.

  9. A roadmap for the integration of culture into developmental psychopathology.

    PubMed

    Causadias, José M

    2013-11-01

    In this paper, I propose a roadmap for the integration of culture in developmental psychopathology. This integration is pressing because culture continues to be somewhat disconnected from theory, research, training, and interventions in developmental psychopathology, thus limiting our understanding of the epigenesis of mental health. I argue that in order to successfully integrate culture into developmental psychopathology, it is crucial to (a) study cultural development, (b) consider both individual-level and social-level cultural processes, (c) examine the interplay between culture and biology, and (d) promote improved and direct cultural assessment. I provide evidence in support of each of these guidelines, present alternative conceptual frameworks, and suggest new lines of research. Hopefully, that these directions will contribute to the emerging field of cultural development and psychopathology, which focuses on the elucidation of the cultural processes that initiate, maintain, or derail trajectories of normal and abnormal behavior.

  10. Toward A Fail-Safe Air Force Culture: Creating a Resilient Future While Avoiding Past Mistakes

    DTIC Science & Technology

    2012-10-01

    process often uses the “ Swiss cheese ” model to evaluate accidents. The image of holes in the protective cheese layers (proactive and reactive measures...minimize the number and size of holes in each slice of cheese . More importantly, however, a HRO’s 16 focus is on “the process of the slices lining up as

  11. Shock Wave Treatment Protects From Neuronal Degeneration via a Toll-Like Receptor 3 Dependent Mechanism: Implications of a First-Ever Causal Treatment for Ischemic Spinal Cord Injury.

    PubMed

    Lobenwein, Daniela; Tepeköylü, Can; Kozaryn, Radoslaw; Pechriggl, Elisabeth J; Bitsche, Mario; Graber, Michael; Fritsch, Helga; Semsroth, Severin; Stefanova, Nadia; Paulus, Patrick; Czerny, Martin; Grimm, Michael; Holfeld, Johannes

    2015-10-27

    Paraplegia following spinal cord ischemia represents a devastating complication of both aortic surgery and endovascular aortic repair. Shock wave treatment was shown to induce angiogenesis and regeneration in ischemic tissue by modulation of early inflammatory response via Toll-like receptor (TLR) 3 signaling. In preclinical and clinical studies, shock wave treatment had a favorable effect on ischemic myocardium. We hypothesized that shock wave treatment also may have a beneficial effect on spinal cord ischemia. A spinal cord ischemia model in mice and spinal slice cultures ex vivo were performed. Treatment groups received immediate shock wave therapy, which resulted in decreased neuronal degeneration and improved motor function. In spinal slice cultures, the activation of TLR3 could be observed. Shock wave effects were abolished in spinal slice cultures from TLR3(-/-) mice, whereas the effect was still present in TLR4(-/-) mice. TLR4 protein was found to be downregulated parallel to TLR3 signaling. Shock wave-treated animals showed significantly better functional outcome and survival. The protective effect on neurons could be reproduced in human spinal slices. Shock wave treatment protects from neuronal degeneration via TLR3 signaling and subsequent TLR4 downregulation. Consequently, it represents a promising treatment option for the devastating complication of spinal cord ischemia after aortic repair. © 2015 The Authors. Published on behalf of the American Heart Association, Inc., by Wiley Blackwell.

  12. OXIDATION OF POLYCHLORINATED BIPHENYLS BY LIVER TISSUE SLICES FROM PHENOBARBITAL-PRETREATED MICE IS CONGENER-SPECIFIC AND ATROPSELECTIVE

    PubMed Central

    Wu, Xianai; Duffel, Michael; Lehmler, Hans-Joachim

    2013-01-01

    Mouse models are powerful tools to study the developmental neurotoxicity of polychlorinated biphenyls (PCBs); however, studies of the oxidation of chiral PCB congeners to potentially neurotoxic hydroxylated metabolites (OH-PCBs) in mice have not been reported. Here we investigate the atropselective oxidation of chiral PCB 91 (2,2',3,4',6-pentachlorobiphenyl), PCB 95 (2,2',3,5',6-pentachlorobiphenyl), PCB 132 (2,2',3,3',4,6'-hexachlorobiphenyl), PCB 136 (2,2',3,3',6,6'-hexachlorobiphenyl) and PCB 149 (2,2',3,4',5',6-hexachlorobiphenyl) to OH-PCBs in liver tissue slices prepared from female mice. The metabolite profile of PCB 136 typically followed the rank order 5-OH-PCB > 4-OH-PCB > 4,5-OH-PCB, and metabolite levels increased with PCB concentration and incubation time. A similar OH-PCB profile was observed with the other PCB congeners, with 5-OH-PCB:4-OH-PCB ratios ranging from 2 to 12. More 5-OH-PCB 136 was formed in liver tissue slices obtained from animals pretreated with phenobarbital (P450 2B inducer) or, to a lesser extent, dexamethasone (P450 2B and 3A enzyme inducer) compared to tissue slices prepared from vehicle-pretreated animals. The apparent rate of 5-OH-PCBs formation followed the approximate rank order PCB 149 > PCB 91 > PCB 132 ~ PCB 136 > PCB 95. Atropselective gas chromatography revealed a congener-specific atropisomeric enrichment of major OH-PCB metabolites. Comparison of our results with published OH-PCB patterns and chiral signatures (i.e., the direction and extent of the atropisomeric enrichment) from rat liver microsomal revealed drastic differences between both species, especially following induction of P450 2B enzymes. These species differences in the metabolism of chiral PCBs should be considered in developmental neurotoxicity studies of PCBs. PMID:24107130

  13. Culture and Development: A Systematic Relationship.

    PubMed

    Keller, Heidi

    2017-09-01

    This article argues that the relationships between culture and development are differential and systematic. Therefore the presentation of the Western middle-class developmental pathway in textbooks as universal is grossly neglecting the reality and the psychologies of the majority of the world' s population. First, the conception of culture as the representation of environmental conditions is presented. The level of formal education acts as organizer of social milieus that define different learning environments for children. Mainly two developmental pathways are portrayed: the Western middle-class trajectory and the traditional farmer childhood. Different developmental principles are highlighted, demonstrating systematic cultural differences in the development of a conception of the self: developmental dynamics as exemplified in early mother infant interactions, the timing of developmental milestones emphasizing cultural precocities in motor development and self-recognition, developmental gestalts in different attachment relationships and precursors and consequences demonstrating that different, sometimes contradictory behavioral patterns have the same developmental consequences with the examples of empathy development and autobiographical memory. It is argued that evaluating the development in one pathway with the principles and standards of the other is unscientific and unethical. The recognition of different developmental pathways is a necessity for basic science and a moral obligation for the applied fields.

  14. Ex-vivo imaging of excised tissue using vital dyes and confocal microscopy

    PubMed Central

    Johnson, Simon; Rabinovitch, Peter

    2012-01-01

    Vital dyes routinely used for staining cultured cells can also be used to stain and image live tissue slices ex-vivo. Staining tissue with vital dyes allows researchers to collect structural and functional data simultaneously and can be used for qualitative or quantitative fluorescent image collection. The protocols presented here are useful for structural and functional analysis of viable properties of cells in intact tissue slices, allowing for the collection of data in a structurally relevant environment. With these protocols, vital dyes can be applied as a research tool to disease processes and properties of tissue not amenable to cell culture based studies. PMID:22752953

  15. The concentration-dependent effect of progesterone on follicle growth in the mouse ovary.

    PubMed

    Komatsu, Kouji; Masubuchi, Satoru

    2017-06-21

    Follicle growth in the mammalian ovary is coordinately controlled by multiple factors to sustain periodic ovulation. In this study, we investigated the role of progesterone on follicle growth in the mouse ovary. As the concentration of progesterone changes during the estrus cycle, we cultured the sliced mouse ovary in a medium containing 10 ng/ml, 100 ng/ml, and 1 μg/ml progesterone. Progesterone promoted the growth of primordial to primary follicles at 100 ng/ml, while it suppressed the growth of secondary follicles at 1 μg/ml. Follicles at other developmental stages in the cultured ovary were unaffected with different concentrations of progesterone. The number of ovulated oocytes increased in the medium containing 100 ng/ml progesterone but decreased in the presence of 1 μg/ml progesterone. Follicles expressed two types of progesterone receptors, progesterone receptor (PGR) and PGR membrane component 1 (PGRMC1). While PGR shows transient expression on granulosa cells of Graafian follicles, PGRMC1 expresses in granulosa cells of developing follicles. These results suggest that progesterone controls the growth of developing follicles through PGRMC1. Our study shows that the effect of progesterone on ovulation and follicle growth in mouse ovary is dependent on the concentration of progesterone and the follicle stage.

  16. Rosiglitazone Suppresses In Vitro Seizures in Hippocampal Slice by Inhibiting Presynaptic Glutamate Release in a Model of Temporal Lobe Epilepsy.

    PubMed

    Wong, Shi-Bing; Cheng, Sin-Jhong; Hung, Wei-Chen; Lee, Wang-Tso; Min, Ming-Yuan

    2015-01-01

    Peroxisomal proliferator-activated receptor gamma (PPARγ) is a nuclear hormone receptor whose agonist, rosiglitazone has a neuroprotective effect to hippocampal neurons in pilocarpine-induced seizures. Hippocampal slice preparations treated in Mg2+ free medium can induce ictal and interictal-like epileptiform discharges, which is regarded as an in vitro model of N-methyl-D-aspartate (NMDA) receptor-mediated temporal lobe epilepsy (TLE). We applied rosiglitazone in hippocampal slices treated in Mg2+ free medium. The effects of rosiglitazone on hippocampal CA1-Schaffer collateral synaptic transmission were tested. We also examined the neuroprotective effect of rosiglitazone toward NMDA excitotoxicity on cultured hippocampal slices. Application of 10 μM rosiglitazone significantly suppressed amplitude and frequency of epileptiform discharges in CA1 neurons. Pretreatment with the PPARγ antagonist GW9662 did not block the effect of rosiglitazone on suppressing discharge frequency, but reverse the effect on suppressing discharge amplitude. Application of rosiglitazone suppressed synaptic transmission in the CA1-Schaffer collateral pathway. By miniature excitatory-potential synaptic current (mEPSC) analysis, rosiglitazone significantly suppressed presynaptic neurotransmitter release. This phenomenon can be reversed by pretreating PPARγ antagonist GW9662. Also, rosiglitazone protected cultured hippocampal slices from NMDA-induced excitotoxicity. The protective effect of 10 μM rosiglitazone was partially antagonized by concomitant high dose GW9662 treatment, indicating that this effect is partially mediated by PPARγ receptors. In conclusion, rosiglitazone suppressed NMDA receptor-mediated epileptiform discharges by inhibition of presynaptic neurotransmitter release. Rosiglitazone protected hippocampal slice from NMDA excitotoxicity partially by PPARγ activation. We suggest that rosiglitazone could be a potential agent to treat patients with TLE.

  17. Precision-cut liver slices of Atlantic cod (Gadus morhua): an in vitro system for studying the effects of environmental contaminants.

    PubMed

    Eide, M; Karlsen, O A; Kryvi, H; Olsvik, P A; Goksøyr, A

    2014-08-01

    The Atlantic cod (Gadus morhua) is an economically important species commonly consumed by humans. The widespread distribution of cod in the North Atlantic Ocean makes it vulnerable to effluents from human activities, such as coastal industries and offshore petroleum exploration. It has been demonstrated that many effluents have adverse effects on cod reproduction and health, e.g. by disrupting endocrine signaling pathways. The liver, expressing important components of the biotransformation and the endocrine system, is one of the main target organs. Thus, reliable and reproducible in vitro systems of the liver are important for studying effects of environmental contaminants. The aim of this study was to investigate precision-cut liver slices (PCLS) as an alternative in vitro system for toxicological studies of the Atlantic cod liver. Slices of 8 mm in diameter and 250 μm thickness were prepared and cultivated from immature cod. Several analyses to measure the liver slice viability were performed: enzyme assays, histology, and morphometric analysis, all confirming cell viability for up to 72 h in culture. The liver slices were also exposed to two well-known model environmental contaminants, β-naphthoflavone (BNF) and 17α-ethynylestradiol (EE2), representing established agonists for the aryl hydrocarbon receptor (AHR) and the estrogen receptor (ER), respectively. The results showed increased transcription of the target genes cytochrome P450 1A (CYP1A) and vitellogenin (VTG), both well-established biomarkers for exposure of fish to the selected compounds. In conclusion, PCLS is a promising in vitro system for toxicological studies of cod liver cells. The liver slices are viable in culture for several days and respond to environmental contaminants in a dose- and time-specific manner. Copyright © 2013 Elsevier B.V. All rights reserved.

  18. Correlation between in vivo and in vitro pulmonary responses to jet propulsion fuel-8 using precision-cut lung slices and a dynamic organ culture system.

    PubMed

    Hays, Allison M; Lantz, R Clark; Witten, Mark L

    2003-01-01

    In tissue slice models, interactions between the heterogeneous cell types comprising the lung parenchyma are maintained thus providing a controlled system for the study of pulmonary toxicology in vitro. However, validation of the model in vitro system must be affirmed. Previous reports, in in vivo systems, have demonstrated that Clara cells and alveolar type II cells are the targets following inhalation of JP-8 jet fuel. We have utilized the lung slice model to determine if cellular targets are similar following in vitro exposure to JP-8. Agar-filled adult rat lung explants were cored and precision cut, using the Brende/Vitron tissue slicer. Slices were cultured on titanium screens located as half-cylinders in cylindrical Teflon cradles that were loaded into standard scintillation vials and incubated at 37 degrees C. Slices were exposed to JP-8 jet fuel (0.5 mg/ml, 1.0 mg/ml, and 1.5 mg/ml in medium) for up to 24 hours. We determined ATP content using a luciferin-luciferase bioluminescent assay. No significant difference was found between the JP-8 jet fuel doses or time points, when compared to controls. Results were correlated with structural alterations following aerosol inhalation of JP-8. As a general observation, ultrastructural evaluation of alveolar type cells revealed an apparent increase in the number and size of surfactant secreting lamellar bodies that was JP-8 jet fuel-dose dependent. These results are similar to those observed following aerosol inhalation exposure. Thus, the lung tissue slice model appears to mimic in vivo effects of JP-8 and therefore is a useful model system for studying the mechanisms of lunginjury following JP-8 exposure.

  19. Tumor Slice Culture: A New Avatar in Personalized Oncology

    DTIC Science & Technology

    2017-09-01

    of the Army position, policy or decision unless so designated by other documentation. REPORT DOCUMENTATION PAGE Form Approved OMB No. 0704-0188...significantly correlated with pathologic assessment of tumor viability/necrosis. We continue to optimize the conditions for TSC by manipulating growth...following exposure to treatments. We are in the process of assessing the pathologic outcome using H&E staining of the treated slices and correlating the

  20. Tumor Slice Culture: A New Avatar in Personalized Oncology

    DTIC Science & Technology

    2017-09-01

    the Army position, policy or decision unless so designated by other documentation. REPORT DOCUMENTATION PAGE Form Approved OMB No. 0704-0188 Public... correlated with pathologic assessment of tumor viability/necrosis. We continue to optimize the conditions for TSC by manipulating growth factors...exposure to treatments. We are in the process of assessing the pathologic outcome using H&E staining of the treated slices and correlating the results

  1. Slice cultures of the imprinting-relevant forebrain area MNH of the domestic chick: quantitative characterization of neuronal morphology.

    PubMed

    Hofmann, H; Braun, K

    1995-05-26

    The persistence of morphological features of neurons in slice cultures of the imprinting-relevant forebrain area MNH (mediorostral neostriatum and hyperstriatum ventrale) of the domestic chick was analysed at 7, 14, 21 and 28 days in vitro. After having been explanted and kept in culture the neurons in vitro have larger soma areas, longer and more extensively branched dendritic trees and lower spine frequencies compared to the neurons in vivo. During the analyzed culturing period, the parameters soma area, total and mean dendritic length, number of dendrites, number of dendritic nodes per dendrite and per neuron as well as the spine densities in different dendritic segments showed no significant differences between early and late periods. Highly correlated in every age group were the total dendritic length and the number of dendritic nodes per neuron, indicating regular ramification during dendritic growth. Since these morphological parameters remain stable during the first 4 weeks in vitro, this culture system may provide a suitable model to investigate experimentally induced morphological changes.

  2. Investigating Functional Regeneration in Organotypic Spinal Cord Co-cultures Grown on Multi-electrode Arrays.

    PubMed

    Heidemann, Martina; Streit, Jürg; Tscherter, Anne

    2015-09-23

    Adult higher vertebrates have a limited potential to recover from spinal cord injury. Recently, evidence emerged that propriospinal connections are a promising target for intervention to improve functional regeneration. So far, no in vitro model exists that grants the possibility to examine functional recovery of propriospinal fibers. Therefore, a representative model that is based on two organotypic spinal cord sections of embryonic rat, cultured next to each other on multi-electrode arrays (MEAs) was developed. These slices grow and, within a few days in vitro, fuse along the sides facing each other. The design of the used MEAs permits the performance of lesions with a scalpel blade through this fusion site without inflicting damage on the MEAs. The slices show spontaneous activity, usually organized in network activity bursts, and spatial and temporal activity parameters such as the location of burst origins, speed and direction of their propagation and latencies between bursts can be characterized. Using these features, it is also possible to assess functional connection of the slices by calculating the amount of synchronized bursts between the two sides. Furthermore, the slices can be morphologically analyzed by performing immunohistochemical stainings after the recordings. Several advantages of the used techniques are combined in this model: the slices largely preserve the original tissue architecture with intact local synaptic circuitry, the tissue is easily and repeatedly accessible and neuronal activity can be detected simultaneously and non-invasively in a large number of spots at high temporal resolution. These features allow the investigation of functional regeneration of intraspinal connections in isolation in vitro in a sophisticated and efficient way.

  3. The use of three-dimensional printing to produce in vitro slice chambers

    PubMed Central

    Hyde, James; MacNicol, Melanie; Odle, Angela; Garcia-Rill, Edgar

    2014-01-01

    Background In recent years, 3D printing technology has become inexpensive and simple enough that any lab can own and use one of these printers. New Method We explored the potential use of 3D printers for quickly and easily producing in vitro slice chambers for patch clamp electrophysiology. Slice chambers were produced using five available plastics: ABS, PLA, Nylon 618, Nylon 680, and T-glase. These “lab-made” chambers were also made using stereolithography through a professional printing service (Shapeways). This study measured intrinsic membrane properties of neurons in the brain stem pedunculopontine nucleus (PPN) and layer V pyramidal neurons in retrosplenial cortex. Results Nylon 680 and T-glase significantly hyperpolarized PPN neurons. ABS increased input resistance, decreased action potential amplitude, and increased firing frequency in pyramidal cortical neurons. To test long term exposure to each plastic, human neuroblastoma SHSY5Y cell cultures were exposed to each plastic for 1 week. ABS decreased cell counts while Nylon 618 and Shapeways plastics eliminated cells. Primary mouse pituitary cultures were also tested for 24-hour exposure. ABS decreased cell counts while Nylon 618 and Shapeways plastics decreased cell counts. Comparison to Existing Methods Chambers can be quickly and inexpensively printed in the lab. ABS, PLA, Nylon 680, and T-glase plastics would suffice for many experiments instead of commercially produced slice chambers. Conclusions While these technologies are still in their infancy, they represent a powerful addition to the lab environment. With careful selection of print material, slice chambers can be quickly and inexpensively manufactured in the lab. PMID:25251556

  4. Group III mGluR regulation of synaptic transmission at the SC-CA1 synapse is developmentally regulated

    PubMed Central

    Ayala, Jennifer E.; Niswender, Colleen M.; Luo, Qingwei; Banko, Jessica L.; Conn, P. Jeffrey

    2008-01-01

    Summary Group III metabotropic glutamate receptors (mGluRs) reduce synaptic transmission at the Schaffer collateral-CA1 (SC-CA1) synapse in rats by a presynaptic mechanism. Previous studies show that low concentrations of the group III-selective agonist, L-AP4, reduce synaptic transmission in slices from neonatal but not adult rats, whereas high micromolar concentrations reduce transmission in both age groups. L-AP4 activates mGluRs 4 and 8 at much lower concentrations than those required to activate mGluR7, suggesting that the group III mGluR subtype modulating transmission is a high affinity receptor in neonates and a low affinity receptor in adults. The previous lack of subtype selective ligands has made it difficult to test this hypothesis. We have measured fEPSPs in the presence of novel subtype selective agents to address this question. We show that the effects of L-AP4 can be blocked by LY341495 in both neonates and adults, verifying that these effects are mediated by mGluRs. In addition, the selective mGluR8 agonist, DCPG, has a significant effect in slices from neonatal rats but does not reduce synaptic transmission in adult slices. The mGluR4 selective allosteric potentiator, PHCCC, is unable to potentiate the L-AP4-induced effects at either age. Taken together, our data suggest that group III mGluRs regulate transmission at the SC-CA1 synapse throughout development but there is a developmental regulation of the subtypes involved so that that both mGluR8 serves this role in neonates but not adults whereas mGluR7 is involved in regulating transmission at this synapse in throughout postnatal development. PMID:18255102

  5. Discovery of Hyperpolarized Molecular Imaging Biomarkers in a Novel Prostate Tissue Slice Culture Model

    DTIC Science & Technology

    2013-06-01

    research is to optimize an MRS-compatible, 3D Tissue Culture Bioreactor for use with primary human prostate tissue cultures (TSCs) and use it to...Tissue Culture Bioreactor ” to be submitted to the Journal Magnetic Resonance in Medicine. CONCLUSIONS: We have engineered a robust MR compatible 3D ...loss of structure, function or metabolism within a NMR compatible 3-D tissue culture bioreactor , and that magnetic resonance spectroscopy studies of

  6. Using Popular Culture in Developmental Writing

    ERIC Educational Resources Information Center

    Barnes, Sharon L.

    2006-01-01

    Using popular culture in my developmental writing course has prompted me to reconsider what it means to create successful developmental writing assignments. Having slipped into the questionable habit of assuming that removing complexity makes an assignment appropriate for developing writers, I pared down a fairly open-ended "media…

  7. Mesenchymal stem cells support neuronal fiber growth in an organotypic brain slice co-culture model.

    PubMed

    Sygnecka, Katja; Heider, Andreas; Scherf, Nico; Alt, Rüdiger; Franke, Heike; Heine, Claudia

    2015-04-01

    Mesenchymal stem cells (MSCs) have been identified as promising candidates for neuroregenerative cell therapies. However, the impact of different isolation procedures on the functional and regenerative characteristics of MSC populations has not been studied thoroughly. To quantify these differences, we directly compared classically isolated bulk bone marrow-derived MSCs (bulk BM-MSCs) to the subpopulation Sca-1(+)Lin(-)CD45(-)-derived MSCs(-) (SL45-MSCs), isolated by fluorescence-activated cell sorting from bulk BM-cell suspensions. Both populations were analyzed with respect to functional readouts, that are, frequency of fibroblast colony forming units (CFU-f), general morphology, and expression of stem cell markers. The SL45-MSC population is characterized by greater morphological homogeneity, higher CFU-f frequency, and significantly increased nestin expression compared with bulk BM-MSCs. We further quantified the potential of both cell populations to enhance neuronal fiber growth, using an ex vivo model of organotypic brain slice co-cultures of the mesocortical dopaminergic projection system. The MSC populations were cultivated underneath the slice co-cultures without direct contact using a transwell system. After cultivation, the fiber density in the border region between the two brain slices was quantified. While both populations significantly enhanced fiber outgrowth as compared with controls, purified SL45-MSCs stimulated fiber growth to a larger degree. Subsequently, we analyzed the expression of different growth factors in both cell populations. The results show a significantly higher expression of brain-derived neurotrophic factor (BDNF) and basic fibroblast growth factor in the SL45-MSCs population. Altogether, we conclude that MSC preparations enriched for primary MSCs promote neuronal regeneration and axonal regrowth, more effectively than bulk BM-MSCs, an effect that may be mediated by a higher BDNF secretion.

  8. Hyoid bone development: An assessment of optimal CT scanner parameters and 3D volume rendering techniques

    PubMed Central

    Cotter, Meghan M.; Whyms, Brian J.; Kelly, Michael P.; Doherty, Benjamin M.; Gentry, Lindell R.; Bersu, Edward T.; Vorperian, Houri K.

    2015-01-01

    The hyoid bone anchors and supports the vocal tract. Its complex shape is best studied in three dimensions, but it is difficult to capture on computed tomography (CT) images and three-dimensional volume renderings. The goal of this study was to determine the optimal CT scanning and rendering parameters to accurately measure the growth and developmental anatomy of the hyoid and to determine whether it is feasible and necessary to use these parameters in the measurement of hyoids from in vivo CT scans. Direct linear and volumetric measurements of skeletonized hyoid bone specimens were compared to corresponding CT images to determine the most accurate scanning parameters and three-dimensional rendering techniques. A pilot study was undertaken using in vivo scans from a retrospective CT database to determine feasibility of quantifying hyoid growth. Scanning parameters and rendering technique affected accuracy of measurements. Most linear CT measurements were within 10% of direct measurements; however, volume was overestimated when CT scans were acquired with a slice thickness greater than 1.25 mm. Slice-by-slice thresholding of hyoid images decreased volume overestimation. The pilot study revealed that the linear measurements tested correlate with age. A fine-tuned rendering approach applied to small slice thickness CT scans produces the most accurate measurements of hyoid bones. However, linear measurements can be accurately assessed from in vivo CT scans at a larger slice thickness. Such findings imply that investigation into the growth and development of the hyoid bone, and the vocal tract as a whole, can now be performed using these techniques. PMID:25810349

  9. Hyoid Bone Development: An Assessment Of Optimal CT Scanner Parameters and Three-Dimensional Volume Rendering Techniques.

    PubMed

    Cotter, Meghan M; Whyms, Brian J; Kelly, Michael P; Doherty, Benjamin M; Gentry, Lindell R; Bersu, Edward T; Vorperian, Houri K

    2015-08-01

    The hyoid bone anchors and supports the vocal tract. Its complex shape is best studied in three dimensions, but it is difficult to capture on computed tomography (CT) images and three-dimensional volume renderings. The goal of this study was to determine the optimal CT scanning and rendering parameters to accurately measure the growth and developmental anatomy of the hyoid and to determine whether it is feasible and necessary to use these parameters in the measurement of hyoids from in vivo CT scans. Direct linear and volumetric measurements of skeletonized hyoid bone specimens were compared with corresponding CT images to determine the most accurate scanning parameters and three-dimensional rendering techniques. A pilot study was undertaken using in vivo scans from a retrospective CT database to determine feasibility of quantifying hyoid growth. Scanning parameters and rendering technique affected accuracy of measurements. Most linear CT measurements were within 10% of direct measurements; however, volume was overestimated when CT scans were acquired with a slice thickness greater than 1.25 mm. Slice-by-slice thresholding of hyoid images decreased volume overestimation. The pilot study revealed that the linear measurements tested correlate with age. A fine-tuned rendering approach applied to small slice thickness CT scans produces the most accurate measurements of hyoid bones. However, linear measurements can be accurately assessed from in vivo CT scans at a larger slice thickness. Such findings imply that investigation into the growth and development of the hyoid bone, and the vocal tract as a whole, can now be performed using these techniques. © 2015 Wiley Periodicals, Inc.

  10. Parenting Practices in Cultural Context: An Ecological Perspective

    ERIC Educational Resources Information Center

    Zarnegar, Zohreh

    2015-01-01

    Despite general consensus that parenting practices influence the developmental processes of children, many questions about the impacts of parenting practices on child development within the cultural context remain unanswered. This article presents how cultural templates influence parenting practices and developmental processes of young children.…

  11. Ecological Human Brain and Young Children's "Naturalist Intelligence" from the Perspective of Developmentally and Culturally Appropriate Practice (DCAP).

    ERIC Educational Resources Information Center

    Hyun, Eunsook

    Based on the view that young children have a different intellectual culture from adults' in the way they know and understand nature, this paper explores ecological human brain development, children's intellectual culture of naturalist intelligence, and developmentally and culturally congruent curricula for young children. The paper discusses the…

  12. The Environmental Neurotoxicant PCB 95 Promotes Synaptogenesis via Ryanodine Receptor-Dependent miR132 Upregulation

    PubMed Central

    Lesiak, Adam; Zhu, Mingyan; Chen, Hao; Appleyard, Suzanne M.; Impey, Soren; Wayman, Gary A.

    2014-01-01

    Non–dioxin-like (NDL) polychlorinated biphenyls (PCBs) are widespread environmental contaminants linked to neuropsychological dysfunction in children. NDL PCBs increase spontaneous Ca2+ oscillations in neurons by stabilizing ryanodine receptor (RyR) calcium release channels in the open configuration, which results in CREB-dependent dendritic outgrowth. In this study, we address the question of whether activation of CREB by NDL PCBs also triggers dendritic spine formation. Nanomolar concentrations of PCB 95, a NDL congener with potent RyR activity, significantly increased spine density and the frequency of miniature EPSCs in primary dissociated rat hippocampal cultures coincident with upregulation of miR132. Inhibition of RyR, CREB, or miR132 as well as expression of a mutant p250GAP cDNA construct that is not suppressed by miR132 blocked PCB 95 effects on spines and miniature EPSCs. PCB 95 also induced spine formation via RyR- and miR132-dependent mechanisms in hippocampal slice cultures. These data demonstrate a novel mechanism of PCB developmental neurotoxicity whereby RyR sensitization modulates spine formation and synaptogenesis via CREB-mediated miR132 upregulation, which in turn suppresses the translation of p250GAP, a negative regulator of synaptogenesis. In light of recent evidence implicating miR132 dysregulation in Rett syndrome and schizophrenia, these findings identify NDL PCBs as potential environmental risk factors for neurodevelopmental disorders. PMID:24431430

  13. Antibody uptake into neurons occurs primarily via clathrin-dependent Fcγ receptor endocytosis and is a prerequisite for acute tau protein clearance.

    PubMed

    Congdon, Erin E; Gu, Jiaping; Sait, Hameetha B R; Sigurdsson, Einar M

    2013-12-06

    Tau immunotherapy is effective in transgenic mice, but the mechanisms of Tau clearance are not well known. To this end, Tau antibody uptake was analyzed in brain slice cultures and primary neurons. Internalization was rapid (<1 h), saturable, and substantial compared with control mouse IgG. Furthermore, temperature reduction to 4 °C, an excess of unlabeled mouse IgG, or an excess of Tau antibodies reduced uptake in slices by 63, 41, and 62%, respectively (p = 0.002, 0.04, and 0.005). Uptake strongly correlated with total and insoluble Tau levels (r(2) = 0.77 and 0.87 and p = 0.002 and 0.0002), suggesting that Tau aggregates influence antibody internalization and/or retention within neurons. Inhibiting phagocytosis did not reduce uptake in slices or neuronal cultures, indicating limited microglial involvement. In contrast, clathrin-specific inhibitors reduced uptake in neurons (≤ 78%, p < 0.0001) and slices (≤ 35%, p = 0.03), demonstrating receptor-mediated endocytosis as the primary uptake pathway. Fluid phase endocytosis accounted for the remainder of antibody uptake in primary neurons, based on co-staining with internalized dextran. The receptor-mediated uptake is to a large extent via low affinity FcγII/III receptors and can be blocked in slices (43%, p = 0.04) and neurons (53%, p = 0.008) with an antibody against these receptors. Importantly, antibody internalization appears to be necessary for Tau reduction in primary neurons. Overall, these findings clarify that Tau antibody uptake is primarily receptor-mediated, that these antibodies are mainly found in neurons with Tau aggregates, and that their intracellular interaction leads to clearance of Tau pathology, all of which have major implications for therapeutic development of this approach.

  14. De Novo Synthesized Estradiol Protects against Methylmercury-Induced Neurotoxicity in Cultured Rat Hippocampal Slices

    PubMed Central

    Ishihara, Yasuhiro; Komatsu, Shota; Munetsuna, Eiji; Onizaki, Masahiro; Ishida, Atsuhiko; Kawato, Suguru; Mukuda, Takao

    2013-01-01

    Background Estrogen, a class of female sex steroids, is neuroprotective. Estrogen is synthesized in specific areas of the brain. There is a possibility that the de novo synthesized estrogen exerts protective effect in brain, although direct evidence for the neuroprotective function of brain-synthesized estrogen has not been clearly demonstrated. Methylmercury (MeHg) is a neurotoxin that induces neuronal degeneration in the central nervous system. The neurotoxicity of MeHg is region-specific, and the molecular mechanisms for the selective neurotoxicity are not well defined. In this study, the protective effect of de novo synthesized 17β-estradiol on MeHg-induced neurotoxicity in rat hippocampus was examined. Methodology/Principal Findings Neurotoxic effect of MeHg on hippocampal organotypic slice culture was quantified by propidium iodide fluorescence imaging. Twenty-four-hour treatment of the slices with MeHg caused cell death in a dose-dependent manner. The toxicity of MeHg was attenuated by pre-treatment with exogenously added estradiol. The slices de novo synthesized estradiol. The estradiol synthesis was not affected by treatment with 1 µM MeHg. The toxicity of MeHg was enhanced by inhibition of de novo estradiol synthesis, and the enhancement of toxicity was recovered by the addition of exogenous estradiol. The neuroprotective effect of estradiol was inhibited by an estrogen receptor (ER) antagonist, and mimicked by pre-treatment of the slices with agonists for ERα and ERβ, indicating the neuroprotective effect was mediated by ERs. Conclusions/Significance Hippocampus de novo synthesized estradiol protected hippocampal cells from MeHg-induced neurotoxicity via ERα- and ERβ-mediated pathways. The self-protective function of de novo synthesized estradiol might be one of the possible mechanisms for the selective sensitivity of the brain to MeHg toxicity. PMID:23405170

  15. Baicalin increases developmental competence of mouse embryos in vitro by inhibiting cellular apoptosis and modulating HSP70 and DNMT expression

    PubMed Central

    QI, Xiaonan; LI, Huatao; CONG, Xia; WANG, Xin; JIANG, Zhongling; CAO, Rongfeng; TIAN, Wenru

    2016-01-01

    Scutellaria baicalensis has been effectively used in Chinese traditional medicine to prevent miscarriages. However, little information is available on its mechanism of action. This study is designed specifically to reveal how baicalin, the main effective ingredient of S. baicalensis, improves developmental competence of embryos in vitro, using the mouse as a model. Mouse pronuclear embryos were cultured in KSOM medium supplemented with (0, 2, 4 and 8 μg/ml) baicalin. The results demonstrated that in vitro culture conditions significantly decreased the blastocyst developmental rate and blastocyst quality, possibly due to increased cellular stress and apoptosis. Baicalin (4 µg/ml) significantly increased 2- and 4-cell cleavage rates, morula developmental rate, and blastocyst developmental rate and cell number of in vitro-cultured mouse embryos. Moreover, baicalin increased the expression of Gja1, Cdh1, Bcl-2, and Dnmt3a genes, decreased the expression of Dnmt1 gene, and decreased cellular stress and apoptosis as it decreased the expression of HSP70, CASP3, and BAX and increased BCL-2 expression in blastocysts cultured in vitro. In conclusion, baicalin improves developmental competence of in vitro-cultured mouse embryos through inhibition of cellular apoptosis and HSP70 expression, and improvement of DNA methylation. PMID:27478062

  16. A new extracellular multirecording system for electrophysiological studies: application to hippocampal organotypic cultures.

    PubMed

    Stoppini, L; Duport, S; Corrèges, P

    1997-03-01

    The present paper describes a new multirecording device which performs continuous electrophysiological studies on organotypic cultures. This device is formed by a card (Physiocard) carrying the culture which is inserted into an electronic module. Electrical activities are recorded by an array of 30 biocompatible microelectrodes which are adjusted into close contact with the upper surface of the slice culture. The microelectrode array is integrated into the card enabling electrical stimulation and recording of neurons over periods ranging from several hours to a few days outside a Faraday cage. Neuronal responses are recorded and analyzed by a dedicated electronic and acquisition chain. A perfusion chamber is contained in the card, allowing continuous perfusion in sterile conditions. Electrophysiological extracellular recordings and some drugs' effects obtained with this system in hippocampal slice cultures were identical to conventional electrophysiological set-up results with tetrodotoxin, bicuculline, kainate, dexamethasone and NBQX. The Physiocard system allows new insights for studies on nervous tissue and allows sophisticated approaches to be used quicker and more easily. It could be used for various neurophysiological studies or screening tests such as neural network mapping, nervous recovery, epilepsy, neurotoxicity or neuropharmacology.

  17. Functional Stem Cell Integration into Neural Networks Assessed by Organotypic Slice Cultures.

    PubMed

    Forsberg, David; Thonabulsombat, Charoensri; Jäderstad, Johan; Jäderstad, Linda Maria; Olivius, Petri; Herlenius, Eric

    2017-08-14

    Re-formation or preservation of functional, electrically active neural networks has been proffered as one of the goals of stem cell-mediated neural therapeutics. A primary issue for a cell therapy approach is the formation of functional contacts between the implanted cells and the host tissue. Therefore, it is of fundamental interest to establish protocols that allow us to delineate a detailed time course of grafted stem cell survival, migration, differentiation, integration, and functional interaction with the host. One option for in vitro studies is to examine the integration of exogenous stem cells into an existing active neural network in ex vivo organotypic cultures. Organotypic cultures leave the structural integrity essentially intact while still allowing the microenvironment to be carefully controlled. This allows detailed studies over time of cellular responses and cell-cell interactions, which are not readily performed in vivo. This unit describes procedures for using organotypic slice cultures as ex vivo model systems for studying neural stem cell and embryonic stem cell engraftment and communication with CNS host tissue. © 2017 by John Wiley & Sons, Inc. Copyright © 2017 John Wiley & Sons, Inc.

  18. A fast stimulability screening protocol for neuronal cultures on microelectrode arrays.

    PubMed

    Kapucu, Fikret E; Tanskanen, Jarno M A; Yuan, Yuting; Hyttinen, Jari A K

    2015-01-01

    Microelectrode arrays (MEAs) are used to study the electrical activity in brain slices and neuronal cultures. MEA experiments for the analysis of electrical stimulation responses require the tissue or culture to be prone to stimulation. For brain slices, potential stimulation sites may be directly visible in microscope, in which case the determination of stimulability at those locations is sufficient. In unstructured neuronal cultures, potential stimulation sites may not be known a priori, and spatial stimulability screening should be performed. Considering, e.g., 59 microelectrode sites, each to be stimulated several times, may result in long screening times, unacceptable with a MEA system without an integrated CO2 incubator, or in high stimulation effects on the networks. Here, we describe an implementation of a fast stimulation protocol employing pseudorandom stimulation site switching aiming at alleviating the network effects of the stimulability screening. In this paper, we show the usability of the proposed protocol by first detecting stimulable locations and subsequently apply repeated stimulation on the identified potentially stimulable locations to observe an exemplary neuronal pathway.

  19. Application of Real-Time Fluorescent PCR for Quantitative Assessment of Neospora caninum Infections in Organotypic Slice Cultures of Rat Central Nervous System Tissue

    PubMed Central

    Müller, Norbert; Vonlaufen, Nathalie; Gianinazzi, Christian; Leib, Stephen L.; Hemphill, Andrew

    2002-01-01

    The previously described Nc5-specific PCR test for the diagnosis of Neospora caninum infections was used to develop a quantitative PCR assay which allows the determination of infection intensities within different experimental and diagnostic sample groups. The quantitative PCR was performed by using a dual fluorescent hybridization probe system and the LightCycler Instrument for online detection of amplified DNA. This assay was successfully applied for demonstrating the parasite proliferation kinetics in organotypic slice cultures of rat brain which were infected in vitro with N. caninum tachyzoites. This PCR-based method of parasite quantitation with organotypic brain tissue samples can be regarded as a novel ex vivo approach for exploring different aspects of cerebral N. caninum infection. PMID:11773124

  20. Conversion of Synthetic Aβ to In Vivo Active Seeds and Amyloid Plaque Formation in a Hippocampal Slice Culture Model.

    PubMed

    Novotny, Renata; Langer, Franziska; Mahler, Jasmin; Skodras, Angelos; Vlachos, Andreas; Wegenast-Braun, Bettina M; Kaeser, Stephan A; Neher, Jonas J; Eisele, Yvonne S; Pietrowski, Marie J; Nilsson, K Peter R; Deller, Thomas; Staufenbiel, Matthias; Heimrich, Bernd; Jucker, Mathias

    2016-05-04

    The aggregation of amyloid-β peptide (Aβ) in brain is an early event and hallmark of Alzheimer's disease (AD). We combined the advantages of in vitro and in vivo approaches to study cerebral β-amyloidosis by establishing a long-term hippocampal slice culture (HSC) model. While no Aβ deposition was noted in untreated HSCs of postnatal Aβ precursor protein transgenic (APP tg) mice, Aβ deposition emerged in HSCs when cultures were treated once with brain extract from aged APP tg mice and the culture medium was continuously supplemented with synthetic Aβ. Seeded Aβ deposition was also observed under the same conditions in HSCs derived from wild-type or App-null mice but in no comparable way when HSCs were fixed before cultivation. Both the nature of the brain extract and the synthetic Aβ species determined the conformational characteristics of HSC Aβ deposition. HSC Aβ deposits induced a microglia response, spine loss, and neuritic dystrophy but no obvious neuron loss. Remarkably, in contrast to in vitro aggregated synthetic Aβ, homogenates of Aβ deposits containing HSCs induced cerebral β-amyloidosis upon intracerebral inoculation into young APP tg mice. Our results demonstrate that a living cellular environment promotes the seeded conversion of synthetic Aβ into a potent in vivo seeding-active form. In this study, we report the seeded induction of Aβ aggregation and deposition in long-term hippocampal slice cultures. Remarkably, we find that the biological activities of the largely synthetic Aβ aggregates in the culture are very similar to those observed in vivo This observation is the first to show that potent in vivo seeding-active Aβ aggregates can be obtained by seeded conversion of synthetic Aβ in a living (wild-type) cellular environment. Copyright © 2016 the authors 0270-6474/16/365084-10$15.00/0.

  1. Gravity Vector Changes Induce Alterations in Nervous and Testicular Cells in Cultures and in Testis Slices

    NASA Astrophysics Data System (ADS)

    Uva, B.; Strollo, F.; Ricci, F.; Masini, M. A.

    Cultured astrocytes, neurons and testicular cells (myoid, germ, Sertoli, Leydig cells) as well as rat testes and testes'slices, were subjected to modeled microgravity using a three dimensional Random Positioning Machine (10-6G) for 5min, 30min, 1h, 24h and 32h. Parallel cell cultures and tissues were submitted to hypergravity using an hyperfuge (2.5G) for the same period of time. At the end of the rotations the cultures and tissues were fixed, the tissue was sectioned (5 micron). All the specimens were processed for immunohistochemical identification of microtubules, mitochondria, 3 hydroxysteroid dehydrogenase, 17 hydroxysteroid dehydrogenase, caspase 7, heat shock proteins and identification of DNA fragmentation. At 5min at modeled microgravity and hypergravity, the histology of the cells in culture and the tissues was altered, microtubules and mitochondria were disorganized. Numerous cells underwent apoptosis. Immunostaining for enzymes involved in ion transmembrane transport, as Na+/K+ATPase and cotransporter proteins, and in steroidogenesis diminished or was abolished. At 1h in modeled microgravity or hypergravity, HSPs were expressed and ion transport enzymes as well as steroidogenic enzymes were again immunostainable. These data show that microgravity and hypergravity cause only transient alterations, and tissues and cells in cultures are able to adapt to different gravity conditions.

  2. An integrative review of ethnic and cultural variation in socialization and children's self-regulation.

    PubMed

    LeCuyer, Elizabeth A; Zhang, Yi

    2015-04-01

    To examine the evidence for cross-cultural variation in socialization and children's normative self-regulation, based on a contextual-developmental perspective. Nurses and healthcare workers in multi-cultural societies must understand diversity in socializing influences (including parenting) and in children's behaviour. A contextual-developmental perspective implies that normative cultural and ethnic values will influence socializing processes and behaviour, which in turn will influence children's self-regulation. Integrative review. Studies were located using five major search engines from 1990-2011. Domains of a contextual-developmental perspective and a comprehensive definition of self-regulation assisted the generation of search terms. Selected studies compared at least two ethnic or cultural groups and addressed contextual-developmental domains: (1) culturally specific social values, beliefs, or attitudes; (2) socializing behaviours; and (3) children's normative self-regulation. Eleven studies about children's self-regulation were found to have data consistent with a contextual-developmental perspective. Studies used descriptive correlational or comparative designs with primarily convenience sampling; eight confirmed stated hypotheses, three were exploratory. Findings across studies evidenced coherent patterns of sociocultural influence on children's attention, compliance, delay of gratification, effortful control and executive function. A contextual-developmental perspective provided a useful perspective to examine normative differences in values, socializing behaviours and children's self-regulation. This perspective and these findings are expected to guide future research, to assist nurses and healthcare providers to understand diversity in parenting and children's behaviour. © 2014 John Wiley & Sons Ltd.

  3. Developmental Idealism: The Cultural Foundations of World Development Programs

    PubMed Central

    Thornton, Arland; Dorius, Shawn F.; Swindle, Jeffrey

    2015-01-01

    This paper extends theory and research concerning cultural models of development beyond family and demographic matters to a broad range of additional factors, including government, education, human rights, daily social conventions, and religion. Developmental idealism is a cultural model—a set of beliefs and values—that identifies the appropriate goals of development and the ends for achieving these goals. It includes beliefs about positive cause and effect relationships among such factors as economic growth, educational achievement, health, and political governance, as well as strong values regarding many attributes, including economic growth, education, small families, gender equality, and democratic governance. This cultural model has spread from its origins among the elites of northwest Europe to elites and ordinary people throughout the world. Developmental idealism has become so entrenched in local, national, and global social institutions that it has now achieved a taken-for-granted status among many national elites, academics, development practitioners, and ordinary people around the world. We argue that developmental idealism culture has been a fundamental force behind many cultural clashes within and between societies, and continues to be an important cause of much global social change. We suggest that developmental idealism should be included as a causal factor in theories of human behavior and social change. PMID:26457325

  4. Dentin barrier test with transfected bovine pulp-derived cells.

    PubMed

    Schmalz, G; Schuster, U; Thonemann, B; Barth, M; Esterbauer, S

    2001-02-01

    Growth kinetics of SV40 large T-antigen-transfected bovine pulp-derived cells on dentin were investigated. These cells were used in a dentin barrier test device, and the system was evaluated by testing a set of dental filling materials. Cells (120 cells/mm2) were seeded on dentin slices and incubated for up to 21 days. Cell proliferation was recorded using MTT assay. For cytotoxicity tests 3500 cells/mm2 were seeded on dentin discs, which were then incorporated into the dentin barrier test device. After 72 h preincubation test materials were applied. After a 24 h exposure with or without perfusion of the pulpal part of the test device, cell survival was evaluated using MTT assay. The cells revealed similar growth kinetics on dentin slices and on tissue culture plates. In cytotoxicity tests the cells were more sensitive toward the test materials than previously used three-dimensional cultures of human foreskin fibroblasts and as anticipated from clinical experience. Further improvement is expected by using three-dimensional cultures of pulp-derived cells.

  5. The Constructive Marginal of "Moby-Dick": Ishmael and the Developmental Model of Intercultural Sensitivity

    ERIC Educational Resources Information Center

    Morgan, Jeff

    2011-01-01

    Cultural sensitivity theory is the study of how individuals relate to cultural difference. Using literature to help students prepare for study abroad, instructors could analyze character and trace behavior through a model of cultural sensitivity. Milton J. Bennett has developed such an instrument, The Developmental Model of Intercultural…

  6. A new co-ultramicronized composite including palmitoylethanolamide and luteolin to prevent neuroinflammation in spinal cord injury.

    PubMed

    Paterniti, Irene; Impellizzeri, Daniela; Di Paola, Rosanna; Navarra, Michele; Cuzzocrea, Salvatore; Esposito, Emanuela

    2013-07-23

    It has recently been demonstrated that palmitoylethanolamide (PEA), an endogenous lipid amide belonging to the N-acylethanolamine family, exerts neuroprotection in central nervous system (CNS) pathologies. In recent studies, we have demonstrated that treatment with PEA significantly reduced inflammatory secondary events associated with spinal cord injury (SCI). Since oxidative stress is considered to play an important role in neuroinflammatory disorders, in the present work we studied a new composite, a formulation including PEA and the antioxidant compound luteolin (Lut), subjected to an ultramicronization process, co-ultraPEALut. We investigated the effect of co-ultraPEALut (in the respective fixed doses of 10:1 in mass) in both an ex vivo organotypic spinal cord culture model and an in vivo model of SCI. For the organotypic cultures, spinal cords were prepared from mice at postnatal day 6 and were cut into transverse slices of 400 μm thickness to generate the lumbar organotypic slice cultures. After 7 days of culturing, the slices were mechanically injured onto the center of the slice and the co-ultraPEALut was applied at different concentrations (0.00009, 0.0009 and 0.009 g/l) 1 hour before damage. For in vivo studies, SCI was induced in mice through spinal cord compression by the application of vascular clips (force of 24 g) to the dura via a four-level T5 to T8 laminectomy, and co-ultraPEALut (1 mg/kg ip) was administered at 1 and 6 hours after SCI. At 24 hours after SCI, mice were sacrificed and the spinal cords were collected for further evaluation. Additional animals were treated similarly and sacrificed 10 days after SCI. Pretreatment with co-ultraPEALut significantly reduced cyclooxygenase-2 (COX-2) and inducible nitric oxide synthase (iNOS) expression in a concentration-dependent manner, restored neuronal nitric oxide synthase (nNOS) expression at all three tested concentrations, and protected cells by cell death (MTT assay) in spinal cord organotypic cultures. Moreover, we demonstrated in vivo that co-ultraPEALut 1 mg/kg reduced the severity of trauma induced by compression and improved the motor activity evaluated at 10 days post-injury. The present study demonstrates that the protective effect of PEA on SCI-associated neuroinflammation could be improved by co-ultramicronization with Lut possibly due to its antioxidant properties.

  7. A new co-ultramicronized composite including palmitoylethanolamide and luteolin to prevent neuroinflammation in spinal cord injury

    PubMed Central

    2013-01-01

    Background It has recently been demonstrated that palmitoylethanolamide (PEA), an endogenous lipid amide belonging to the N-acylethanolamine family, exerts neuroprotection in central nervous system (CNS) pathologies. In recent studies, we have demonstrated that treatment with PEA significantly reduced inflammatory secondary events associated with spinal cord injury (SCI). Since oxidative stress is considered to play an important role in neuroinflammatory disorders, in the present work we studied a new composite, a formulation including PEA and the antioxidant compound luteolin (Lut), subjected to an ultramicronization process, co-ultraPEALut. We investigated the effect of co-ultraPEALut (in the respective fixed doses of 10:1 in mass) in both an ex vivo organotypic spinal cord culture model and an in vivo model of SCI. Methods For the organotypic cultures, spinal cords were prepared from mice at postnatal day 6 and were cut into transverse slices of 400 μm thickness to generate the lumbar organotypic slice cultures. After 7 days of culturing, the slices were mechanically injured onto the center of the slice and the co-ultraPEALut was applied at different concentrations (0.00009, 0.0009 and 0.009 g/l) 1 hour before damage. For in vivo studies, SCI was induced in mice through spinal cord compression by the application of vascular clips (force of 24 g) to the dura via a four-level T5 to T8 laminectomy, and co-ultraPEALut (1 mg/kg ip) was administered at 1 and 6 hours after SCI. At 24 hours after SCI, mice were sacrificed and the spinal cords were collected for further evaluation. Additional animals were treated similarly and sacrificed 10 days after SCI. Results Pretreatment with co-ultraPEALut significantly reduced cyclooxygenase-2 (COX-2) and inducible nitric oxide synthase (iNOS) expression in a concentration-dependent manner, restored neuronal nitric oxide synthase (nNOS) expression at all three tested concentrations, and protected cells by cell death (MTT assay) in spinal cord organotypic cultures. Moreover, we demonstrated in vivo that co-ultraPEALut 1 mg/kg reduced the severity of trauma induced by compression and improved the motor activity evaluated at 10 days post-injury. Conclusion The present study demonstrates that the protective effect of PEA on SCI-associated neuroinflammation could be improved by co-ultramicronization with Lut possibly due to its antioxidant properties. PMID:23880066

  8. Neuronal uptake of anti-Hu antibody, but not anti-Ri antibody, leads to cell death in brain slice cultures.

    PubMed

    Greenlee, John E; Clawson, Susan A; Hill, Kenneth E; Wood, Blair; Clardy, Stacey L; Tsunoda, Ikuo; Jaskowski, Troy D; Carlson, Noel G

    2014-09-17

    Anti-Hu and anti-Ri antibodies are paraneoplastic immunoglobulin (Ig)G autoantibodies which recognize cytoplasmic and nuclear antigens present in all neurons. Although both antibodies produce similar immunohistological labeling, they recognize different neuronal proteins. Both antibodies are associated with syndromes of central nervous system dysfunction. However, the neurological deficits associated with anti-Hu antibody are associated with neuronal death and are usually irreversible, whereas neurological deficits in patients with anti-Ri antibody may diminish following tumor removal or immunosuppression. To study the effect of anti-Hu and anti-Ri antibodies on neurons, we incubated rat hippocampal and cerebellar slice cultures with anti-Hu or anti-Ri sera from multiple patients. Cultures were evaluated in real time for neuronal antibody uptake and during prolonged incubation for neuronal death. To test the specificity of anti-Hu antibody cytotoxic effect, anti-Hu serum IgG was incubated with rat brain slice cultures prior to and after adsorption with its target Hu antigen, HuD. We demonstrated that: 1) both anti-Hu and anti-Ri antibodies were rapidly taken up by neurons throughout both cerebellum and hippocampus; 2) antibody uptake occurred in living neurons and was not an artifact of antibody diffusion into dead cells; 3) intracellular binding of anti-Hu antibody produced neuronal cell death, whereas uptake of anti-Ri antibody did not affect cell viability during the period of study; and 4) adsorption of anti-Hu antisera against HuD greatly reduced intraneuronal IgG accumulation and abolished cytotoxicity, confirming specificity of antibody-mediated neuronal death. Both anti-Hu and anti-Ri antibodies were readily taken up by viable neurons in slice cultures, but the two antibodies differed markedly in terms of their effects on neuronal viability. The ability of anti-Hu antibodies to cause neuronal death could account for the irreversible nature of paraneoplastic neurological deficits in patients with this antibody response. Our results raise questions as to whether anti-Ri antibody might initially induce reversible neuronal dysfunction, rather than causing cell death. The ability of IgG antibodies to access and react with intracellular neuronal proteins could have implications for other autoimmune diseases involving the central nervous system.

  9. Long-Term Tissue Culture of Adult Brain and Spleen Slices on Nanostructured Scaffolds.

    PubMed

    Kallendrusch, Sonja; Merz, Felicitas; Bechmann, Ingo; Mayr, Stefan G; Zink, Mareike

    2017-05-01

    Long-term tissue culture of adult mammalian organs is a highly promising approach to bridge the gap between single cell cultures and animal experiments, and bears the potential to reduce in vivo studies. Novel biomimetic materials open up new possibilities to maintain the complex tissue structure in vitro; however, survival times of adult tissues ex vivo are still limited to a few days with established state-of-the-art techniques. Here, it is demonstrated that TiO 2 nanotube scaffolds with specific tissue-tailored characteristics can serve as superior substrates for long-term adult brain and spleen tissue culture. High viability of the explants for at least two weeks is achieved and compared to tissues cultured on standard polytetrafluoroethylene (PTFE) membranes. Histological and immunohistochemical staining and live imaging are used to investigate tissue condition after 5 and 14 d in vitro, while environmental scanning electron microscopy qualifies the interaction with the underlying scaffold. In contrast to tissues cultured on PTFE membranes, enhanced tissue morphology is detected in spleen slices, as well as minor cell death in neuronal tissue, both cultured on nanotube scaffolds. This novel biomimetic tissue model will prove to be useful to address fundamental biological and medical questions from tissue regeneration up to tumor progression and therapeutic approaches. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  10. Cultural pathways through universal development.

    PubMed

    Greenfield, Patricia M; Keller, Heidi; Fuligni, Andrew; Maynard, Ashley

    2003-01-01

    We focus our review on three universal tasks of human development: relationship formation, knowledge acquisition, and the balance between autonomy and relatedness at adolescence. We present evidence that each task can be addressed through two deeply different cultural pathways through development: the pathways of independence and interdependence. Whereas core theories in developmental psychology are universalistic in their intentions, they in fact presuppose the independent pathway of development. Because the independent pathway is therefore well-known in psychology, we focus a large part of our review on empirically documenting the alternative, interdependent pathway for each developmental task. We also present three theoretical approaches to culture and development: the ecocultural, the sociohistorical, and the cultural values approach. We argue that an understanding of cultural pathways through human development requires all three approaches. We review evidence linking values (cultural values approach), ecological conditions (ecocultural approach), and socialization practices (sociohistorical approach) to cultural pathways through universal developmental tasks.

  11. Incubation of curing brines for the production of ready-to-eat, uncured, no-nitrite-or-nitrate-added, ground, cooked and sliced ham.

    PubMed

    Krause, B L; Sebranek, J G; Rust, R E; Mendonca, A

    2011-12-01

    Salt concentration, vegetable juice powder (VJP) concentration and temperature were investigated to determine necessary conditions for incubation of curing brines including VJP and a starter culture containing Staphylococcus carnosus prior to production of naturally cured, no-nitrate/nitrite-added meat products. Subsequently, incubated brines were utilized to produce no-nitrate/nitrite-added sliced ham in which quality characteristics and residual nitrite concentrations were measured to determine feasibility of brine incubation for nitrate conversion prior to injection. Two ham treatments (one with VJP and starter culture; one with pre-converted VJP) and a nitrite-added control were used. No differences (P>0.05) were found for color in the VJP treatments. Control sliced ham was redder after 42 days of storage, retaining significantly (P<0.05) greater a* (redness) than either of the VJP treatments. Residual nitrite concentration was greater (P<0.05) in the control hams during the first week of storage. While the nitrite-added control retained greater red color and initially had more residual nitrite than the VJP treatments, the two VJP treatments did not differ from each other. Copyright © 2011 Elsevier Ltd. All rights reserved.

  12. Alkaloid fraction of Uncaria rhynchophylla protects against N-methyl-D-aspartate-induced apoptosis in rat hippocampal slices.

    PubMed

    Lee, Jongseok; Son, Dongwook; Lee, Pyeongjae; Kim, Sun-Yeou; Kim, Hocheol; Kim, Chang-Ju; Lim, Eunhee

    2003-09-04

    Uncaria rhynchophylla is a medicinal herb which has sedative and anticonvulsive effects and has been applied in the treatment of epilepsy in Oriental medicine. In this study, the effect of alkaloid fraction of U. rhynchophylla against N-methyl-D-aspartate (NMDA)-induced neuronal cell death was investigated. Pretreatment with an alkaloid fraction of U. rhynchophylla for 1 h decreased the degree of neuronal damage induced by NMDA exposure in cultured hippocampal slices and also inhibited NMDA-induced enhanced expressions of apoptosis-related genes such as c-jun, p53, and bax. In the present study, the alkaloid fraction of U. rhynchophylla was shown to have a protective property against NMDA-induced cytotoxicity by suppressing the NMDA-induced apoptosis in rat hippocampal slices.

  13. Effectiveness of a developmental curricular design to graduate culturally competent health practitioners.

    PubMed

    Boggis, Debra

    2012-01-01

    With the goal to facilitate cultural competency development of students enrolled in graduate-level health professional education, this study examined the effectiveness of a curricular program guided by the Intercultural Developmental Continuum (IDC) as measured by the Intercultural Developmental Inventory (IDI). The IDI was administered to 17 occupational therapy (OT) students and a control group of 25 non-OT health professional students upon matriculation into their respective programs of graduate study and again upon completion of 3 years of study. OT students participated in a cultural curricular design guided by the IDC, while the control group participated in cultural study not guided by the IDC. Though OT students did not show a significant change in overall developmental orientation mean scores from pre-test to post-test (t = 0.847, p = 0.41), the results indicate that the designed intercultural curriculum increased intercultural competence among those OT students who began their program with the monocultural mindset of polarization (an "us vs. them" evaluative viewpoint) and moved to the interculturally transitional mindset of minimization (recognizing cultural commonalities and elimination of the "us vs. them" mindset). The control group showed a significant decrease in developmental orientation mean scores at post-test (t = 6.1, p < 0.001). No significant group or group by baseline interaction effects were found when comparing overall post-developmental scores adjusting for baseline (F = 2.4, p = 0.131). The curriculum design as guided by the IDC, though it did not significantly increase overall cultural competency of OT students, appears to have mitigated a decrease in competence. Results suggest that the cultural challenges that students face appear to be considerable and, without targeted, integrated intercultural preparation, can overwhelm new health professionals' intercultural capability.

  14. Hyperpolarized 13C MR Markers of Renal Tumor Aggressiveness

    DTIC Science & Technology

    2014-10-01

    as a biomarker of tumor aggressiveness in a MR compatible 3D cell and tissue culture bioreactor ” to be presented at the ISMRM Workshop on Magnetic... Cell Carcinoma, Hyperpolarized 13C MR, Sub-renal capsule, patient derived tissue slice cultures , bioreactor 3. OVERALL PROJECT SUMMARY: Aim...grade from high grade RCCs using human TSCs cultured in a bioreactor . Aim 2:Identify HP 13C metabolic markers that discriminate low grade from

  15. Assessing the Development of Cross-Cultural Competence in Soldiers

    DTIC Science & Technology

    2010-11-01

    five stages of CQ development based on models from developmental psychology including Piaget’s Model of Cognitive Development ( Piaget , 1985) and...and competence development , the Stage Model of Cognitive Skill Acquisition (Ross et al., 2005), and the Bennett Developmental Model of... developmental stages of proficiency and expertise (see the Stage Model of Cognitive Development ). At this level, cross-cultural competence is highly refined and

  16. Mechanical preconditioning enables electrophysiologic coupling of skeletal myoblast cells to myocardium

    PubMed Central

    Treskes, Philipp; Cowan, Douglas B.; Stamm, Christof; Rubach, Martin; Adelmann, Roland; Wittwer, Thorsten; Wahlers, Thorsten

    2015-01-01

    Objective The effect of mechanical preconditioning on skeletal myoblasts in engineered tissue constructs was investigated to resolve issues associated with conduction block between skeletal myoblast cells and cardiomyocytes. Methods Murine skeletal myoblasts were used to generate engineered tissue constructs with or without application of mechanical strain. After in vitro myotube formation, engineered tissue constructs were co-cultured for 6 days with viable embryonic heart slices. With the use of sharp electrodes, electrical coupling between engineered tissue constructs and embryonic heart slices was assessed in the presence or absence of pharmacologic agents. Results The isolation and expansion procedure for skeletal myoblasts resulted in high yields of homogeneously desmin-positive (97.1% ± 0.1%) cells. Mechanical strain was exerted on myotubes within engineered tissue constructs during gelation of the matrix, generating preconditioned engineered tissue constructs. Electrical coupling between preconditioned engineered tissue constructs and embryonic heart slices was observed; however, no coupling was apparent when engineered tissue constructs were not subjected to mechanical strain. Coupling of cells from engineered tissue constructs to cells in embryonic heart slices showed slower conduction velocities than myocardial cells with the embryonic heart slices (preconditioned engineered tissue constructs vs embryonic heart slices: 0.04 ± 0.02 ms vs 0.10 ± 0.05 ms, P = .011), lower stimulation frequencies (preconditioned engineered tissue constructs vs maximum embryonic heart slices: 4.82 ± 1.42 Hz vs 10.58 ± 1.56 Hz; P = .0009), and higher sensitivities to the gap junction inhibitor (preconditioned engineered tissue constructs vs embryonic heart slices: 0.22 ± 0.07 mmol/L vs 0.93 ± 0.15 mmol/L; P = .0004). Conclusions We have generated skeletal myoblast–based transplantable grafts that electrically couple to myocardium. PMID:22980065

  17. Universal, Developmental, and Variable Aspects of Young Children's Play: A Cross-Cultural Comparison of Pretending at Home.

    ERIC Educational Resources Information Center

    Haight, Wendy L.; Wang, Xiao-lei; Fung, Heidi Han-tih; Williams, Kimberley; Mintz, Judith

    1999-01-01

    This study used longitudinal data from five Irish American families and nine Chinese families in Taiwan, in conjunction with cultural psychology research evidence, to propose universal, culturally variable, and developmental dimensions of children's pretend play. Findings raise the theoretical issue of how universal and variable dimensions of…

  18. Music listening in families and peer groups: benefits for young people's social cohesion and emotional well-being across four cultures

    PubMed Central

    Boer, Diana; Abubakar, Amina

    2014-01-01

    Families are central to the social and emotional development of youth, and most families engage in musical activities together, such as listening to music or talking about their favorite songs. However, empirical evidence of the positive effects of musical family rituals on social cohesion and emotional well-being is scarce. Furthermore, the role of culture in the shaping of musical family rituals and their psychological benefits has been neglected entirely. This paper investigates musical rituals in families and in peer groups (as an important secondary socialization context) in two traditional/collectivistic and two secular/individualistic cultures, and across two developmental stages (adolescence vs. young adulthood). Based on cross-sectional data from 760 young people in Kenya, the Philippines, New Zealand, and Germany, our study revealed that across cultures music listening in families and in peer groups contributes to family and peer cohesion, respectively. Furthermore, the direct contribution of music in peer groups on well-being appears across cultural contexts, whereas musical family rituals affect emotional well-being in more traditional/collectivistic contexts. Developmental analyses show that musical family rituals are consistently and strongly related to family cohesion across developmental stages, whereas musical rituals in peer groups appear more dependent on the developmental stage (in interaction with culture). Contributing to developmental as well as cross-cultural psychology, this research elucidated musical rituals and their positive effects on the emotional and social development of young people across cultures. The implications for future research and family interventions are discussed. PMID:24847296

  19. Music listening in families and peer groups: benefits for young people's social cohesion and emotional well-being across four cultures.

    PubMed

    Boer, Diana; Abubakar, Amina

    2014-01-01

    Families are central to the social and emotional development of youth, and most families engage in musical activities together, such as listening to music or talking about their favorite songs. However, empirical evidence of the positive effects of musical family rituals on social cohesion and emotional well-being is scarce. Furthermore, the role of culture in the shaping of musical family rituals and their psychological benefits has been neglected entirely. This paper investigates musical rituals in families and in peer groups (as an important secondary socialization context) in two traditional/collectivistic and two secular/individualistic cultures, and across two developmental stages (adolescence vs. young adulthood). Based on cross-sectional data from 760 young people in Kenya, the Philippines, New Zealand, and Germany, our study revealed that across cultures music listening in families and in peer groups contributes to family and peer cohesion, respectively. Furthermore, the direct contribution of music in peer groups on well-being appears across cultural contexts, whereas musical family rituals affect emotional well-being in more traditional/collectivistic contexts. Developmental analyses show that musical family rituals are consistently and strongly related to family cohesion across developmental stages, whereas musical rituals in peer groups appear more dependent on the developmental stage (in interaction with culture). Contributing to developmental as well as cross-cultural psychology, this research elucidated musical rituals and their positive effects on the emotional and social development of young people across cultures. The implications for future research and family interventions are discussed.

  20. Oxygen Glucose Deprivation in Rat Hippocampal Slice Cultures Results in Alterations in Carnitine Homeostasis and Mitochondrial Dysfunction

    PubMed Central

    Rau, Thomas F.; Lu, Qing; Sharma, Shruti; Sun, Xutong; Leary, Gregory; Beckman, Matthew L.; Hou, Yali; Wainwright, Mark S.; Kavanaugh, Michael; Poulsen, David J.; Black, Stephen M.

    2012-01-01

    Mitochondrial dysfunction characterized by depolarization of mitochondrial membranes and the initiation of mitochondrial-mediated apoptosis are pathological responses to hypoxia-ischemia (HI) in the neonatal brain. Carnitine metabolism directly supports mitochondrial metabolism by shuttling long chain fatty acids across the inner mitochondrial membrane for beta-oxidation. Our previous studies have shown that HI disrupts carnitine homeostasis in neonatal rats and that L-carnitine can be neuroprotective. Thus, this study was undertaken to elucidate the molecular mechanisms by which HI alters carnitine metabolism and to begin to elucidate the mechanism underlying the neuroprotective effect of L-carnitine (LCAR) supplementation. Utilizing neonatal rat hippocampal slice cultures we found that oxygen glucose deprivation (OGD) decreased the levels of free carnitines (FC) and increased the acylcarnitine (AC): FC ratio. These changes in carnitine homeostasis correlated with decreases in the protein levels of carnitine palmitoyl transferase (CPT) 1 and 2. LCAR supplementation prevented the decrease in CPT1 and CPT2, enhanced both FC and the AC∶FC ratio and increased slice culture metabolic viability, the mitochondrial membrane potential prior to OGD and prevented the subsequent loss of neurons during later stages of reperfusion through a reduction in apoptotic cell death. Finally, we found that LCAR supplementation preserved the structural integrity and synaptic transmission within the hippocampus after OGD. Thus, we conclude that LCAR supplementation preserves the key enzymes responsible for maintaining carnitine homeostasis and preserves both cell viability and synaptic transmission after OGD. PMID:22984394

  1. Conserved pattern of tangential neuronal migration during forebrain development.

    PubMed

    Métin, Christine; Alvarez, Chantal; Moudoux, David; Vitalis, Tania; Pieau, Claude; Molnár, Zoltán

    2007-08-01

    Origin, timing and direction of neuronal migration during brain development determine the distinct organization of adult structures. Changes in these processes might have driven the evolution of the forebrain in vertebrates. GABAergic neurons originate from the ganglionic eminence in mammals and migrate tangentially to the cortex. We are interested in differences and similarities in tangential migration patterns across corresponding telencephalic territories in mammals and reptiles. Using morphological criteria and expression patterns of Darpp-32, Tbr1, Nkx2.1 and Pax6 genes, we show in slice cultures of turtle embryos that early cohorts of tangentially migrating cells are released from the medial ganglionic eminence between stages 14 and 18. Additional populations migrate tangentially from the dorsal subpallium. Large cohorts of tangentially migrating neurons originate ventral to the dorsal ventricular ridge at stage 14 and from the lateral ganglionic eminence from stage 15. Release of GABAergic cells from these regions was investigated further in explant cultures. Tangential migration in turtle proceeds in a fashion similar to mammals. In chimeric slice culture and in ovo graft experiments, the tangentially migrating cells behaved according to the host environment - turtle cells responded to the available cues in mouse slices and mouse cells assumed characteristic migratory routes in turtle brains, indicating highly conserved embryonic signals between these distant species. Our study contributes to the evaluation of theories on the origin of the dorsal cortex and indicates that tangential migration is universal in mammals and sauropsids.

  2. Airway mechanics and methods used to visualize smooth muscle dynamics in vitro.

    PubMed

    Cooper, P R; McParland, B E; Mitchell, H W; Noble, P B; Politi, A Z; Ressmeyer, A R; West, A R

    2009-10-01

    Contraction of airway smooth muscle (ASM) is regulated by the physiological, structural and mechanical environment in the lung. We review two in vitro techniques, lung slices and airway segment preparations, that enable in situ ASM contraction and airway narrowing to be visualized. Lung slices and airway segment approaches bridge a gap between cell culture and isolated ASM, and whole animal studies. Imaging techniques enable key upstream events involved in airway narrowing, such as ASM cell signalling and structural and mechanical events impinging on ASM, to be investigated.

  3. Role of LIGHT in the pathogenesis of joint destruction in rheumatoid arthritis

    PubMed Central

    Sabokbar, Afsie; Afrough, Sara; Mahoney, David J; Uchihara, Yoshinobu; Swales, Catherine; Athanasou, Nicholas A

    2017-01-01

    AIM To characterise the role of substitutes for receptor-activator nuclear factor kappa-B ligand (RANKL) in rheumatoid arthritis (RA) joint destruction. METHODS Synovial fluid (SF) macrophages isolated from the knee joint of RA patients were incubated with 25 ng/mL macrophage-colony stimulating factor (M-CSF) and 50 ng/mL LIGHT (lymphotoxin-like, exhibits inducible expression and competes with herpes simplex virus glycoprotein D for herpes virus entry mediator, a receptor expressed by T lymphocytes) in the presence and absence of 25 ng/mL RANKL and 100 ng/mL osteoprotegerin (OPG) on glass coverslips and dentine slices. Osteoclastogenesis was assessed by the formation of multinucleated cells (MNCs) expressing tartrate-resistant acid phosphatase (TRAP) on coverslips and the extent of lacunar resorption pit formation on dentine slices. The concentration of LIGHT in RA and osteoarthritis (OA) synovial fluid was measured by an enzyme-linked immunosorbent assay (ELISA) and the expression of LIGHT in RA and OA synovium was determined by immunohistochemistry using an indirect immunoperoxidase technique. RESULTS In cultures of RA SF macrophages treated with LIGHT and M-CSF, there was significant formation of TRAP + MNCs on coverslips and extensive lacunar resorption pit formation on dentine slices. SF-macrophage-osteoclast differentiation was not inhibited by the addition of OPG, a decoy receptor for RANKL. Resorption pits were smaller and less confluent than in RANKL-treated cultures but the overall percentage area of the dentine slice resorbed was comparable in LIGHT- and RANKL-treated cultures. LIGHT significantly stimulated RANKL-induced lacunar resorption compared with RA SF macrophages treated with either RANKL or LIGHT alone. LIGHT was strongly expressed by synovial lining cells, subintimal macrophages and endothelial cells in RA synovium and the concentration of LIGHT was much higher in RA compared with OA SF. CONCLUSION LIGHT is highly expressed in RA synovium and SF, stimulates RANKL-independent/dependent osteoclastogenesis from SF macrophages and may contribute to marginal erosion formation. PMID:28589079

  4. DEVELOPMENTAL TOXICITY OF DI- AND TETRACHLOROETHANE AND DICHLOROPROPANE IN EMBRYO CULTURE

    EPA Science Inventory

    DEVELOPMENTAL TOXICITY OF DI- AND TETRACHLOROETHANE AND DICHLOROPROPANE IN EMBRYO CULTURE. JE Andrews, H Nichols, and ES Hunter. Reproductive Toxicology Division, NHEERL, USEPA, RTP, NC.

    Disinfection of drinking water with chlorine results in numerous chlorinated byprodu...

  5. Hippocampal Dendritic Spines Are Segregated Depending on Their Actin Polymerization

    PubMed Central

    Domínguez-Iturza, Nuria; Calvo, María; Benoist, Marion; Esteban, José Antonio; Morales, Miguel

    2016-01-01

    Dendritic spines are mushroom-shaped protrusions of the postsynaptic membrane. Spines receive the majority of glutamatergic synaptic inputs. Their morphology, dynamics, and density have been related to synaptic plasticity and learning. The main determinant of spine shape is filamentous actin. Using FRAP, we have reexamined the actin dynamics of individual spines from pyramidal hippocampal neurons, both in cultures and in hippocampal organotypic slices. Our results indicate that, in cultures, the actin mobile fraction is independently regulated at the individual spine level, and mobile fraction values do not correlate with either age or distance from the soma. The most significant factor regulating actin mobile fraction was the presence of astrocytes in the culture substrate. Spines from neurons growing in the virtual absence of astrocytes have a more stable actin cytoskeleton, while spines from neurons growing in close contact with astrocytes show a more dynamic cytoskeleton. According to their recovery time, spines were distributed into two populations with slower and faster recovery times, while spines from slice cultures were grouped into one population. Finally, employing fast lineal acquisition protocols, we confirmed the existence of loci with high polymerization rates within the spine. PMID:26881098

  6. Human Cerebrospinal Fluid Promotes Neuronal Viability and Activity of Hippocampal Neuronal Circuits In Vitro

    PubMed Central

    Perez-Alcazar, Marta; Culley, Georgia; Lyckenvik, Tim; Mobarrez, Kristoffer; Bjorefeldt, Andreas; Wasling, Pontus; Seth, Henrik; Asztely, Frederik; Harrer, Andrea; Iglseder, Bernhard; Aigner, Ludwig; Hanse, Eric; Illes, Sebastian

    2016-01-01

    For decades it has been hypothesized that molecules within the cerebrospinal fluid (CSF) diffuse into the brain parenchyma and influence the function of neurons. However, the functional consequences of CSF on neuronal circuits are largely unexplored and unknown. A major reason for this is the absence of appropriate neuronal in vitro model systems, and it is uncertain if neurons cultured in pure CSF survive and preserve electrophysiological functionality in vitro. In this article, we present an approach to address how human CSF (hCSF) influences neuronal circuits in vitro. We validate our approach by comparing the morphology, viability, and electrophysiological function of single neurons and at the network level in rat organotypic slice and primary neuronal cultures cultivated either in hCSF or in defined standard culture media. Our results demonstrate that rodent hippocampal slices and primary neurons cultured in hCSF maintain neuronal morphology and preserve synaptic transmission. Importantly, we show that hCSF increases neuronal viability and the number of electrophysiologically active neurons in comparison to the culture media. In summary, our data indicate that hCSF represents a physiological environment for neurons in vitro and a superior culture condition compared to the defined standard media. Moreover, this experimental approach paves the way to assess the functional consequences of CSF on neuronal circuits as well as suggesting a novel strategy for central nervous system (CNS) disease modeling. PMID:26973467

  7. The Effect of Culture Methods and Serum Supplementation on Developmental Competence of Bovine Embryos Cultured In Vitro

    USDA-ARS?s Scientific Manuscript database

    The objective of this study was to compare the developmental competence of bovine in vitro fertilized embryos in three different culture methods; microdrop method (50 µl of medium under mineral oil in petri dishes) compared to tube methods (1 ml of medium in tubes) with or without oil overlay, and t...

  8. NCT and Culture-Conscious Developmental Science

    ERIC Educational Resources Information Center

    Downing-Wilson, Deborah; Pelaprat, Etienne; Rosero, Ivan; Vadeboncoeur, Jennifer; Packer, Martin; Cole, Michael

    2013-01-01

    The authors share the belief that there is great potential for developmental science in bringing the ideas of Niche Construction Theory (NCT), as developed in evolutionary biology, into conversation with Vygotskian-inspired theories such as cultural-historical and activity theories, distributed cognition, and embodied cognition, although from…

  9. Camp Adventure: Bringing A Slice of America to Military Dependents Overseas.

    ERIC Educational Resources Information Center

    Edginton, Christopher R.; Little, Sandra L.

    1988-01-01

    Looks at the mission of Camp Adventure, a U.S. cultural contact summer camp program contracted between the Department of Defense and the University of Oregon for the children of military personnel overseas. (RWB)

  10. Aminochrome decreases NGF, GDNF and induces neuroinflammation in organotypic midbrain slice cultures.

    PubMed

    de Araújo, Fillipe M; Ferreira, Rafael S; Souza, Cleide S; Dos Santos, Cleonice Creusa; Rodrigues, Tácio L R S; E Silva, Juliana Helena C; Gasparotto, Juciano; Gelain, Daniel Pens; El-Bachá, Ramon S; D Costa, Maria de Fátima; Fonseca, José Claudio M; Segura-Aguilar, Juan; Costa, Silvia L; Silva, Victor Diogenes A

    2018-05-01

    Recent evidence shows that aminochrome induces glial activation related to neuroinflammation. This dopamine derived molecule induces formation and stabilization of alpha-synuclein oligomers, mitochondria dysfunction, oxidative stress, dysfunction of proteasomal and lysosomal systems, endoplasmic reticulum stress and disruption of the microtubule network, but until now there has been no evidence of effects on production of cytokines and neurotrophic factors, that are mechanisms involved in neuronal loss in Parkinson's disease (PD). This study examines the potential role of aminochrome on the regulation of NGF, GDNF, TNF-α and IL-1β production and microglial activation in organotypic midbrain slice cultures from P8 - P9 Wistar rats. We demonstrated aminochrome (25 μM, for 24 h) induced reduction of GFAP expression, reduction of NGF and GDNF mRNA levels, morphological changes in Iba1 + cells, and increase of both TNF-α, IL-1β mRNA and protein levels. Moreover, aminochrome (25 μM, for 48 h) induced morphological changes in the edge of slices and reduction of TH expression. These results demonstrate neuroinflammation, as well as negative regulation of neurotrophic factors (GDNF and NGF), may be involved in aminochrome-induced neurodegeneration, and they contribute to a better understanding of PD pathogenesis. Copyright © 2018 Elsevier B.V. All rights reserved.

  11. Music cognition: a developmental perspective.

    PubMed

    Stalinski, Stephanie M; Schellenberg, E Glenn

    2012-10-01

    Although music is universal, there is a great deal of cultural variability in music structures. Nevertheless, some aspects of music processing generalize across cultures, whereas others rely heavily on the listening environment. Here, we discuss the development of musical knowledge, focusing on four themes: (a) capabilities that are present early in development; (b) culture-general and culture-specific aspects of pitch and rhythm processing; (c) age-related changes in pitch perception; and (d) developmental changes in how listeners perceive emotion in music. Copyright © 2012 Cognitive Science Society, Inc.

  12. Multiple and Inseparable: Conceptualizing the Development of Independence and Interdependence

    ERIC Educational Resources Information Center

    Raeff, Catherine

    2006-01-01

    Based on the position that cultural ideologies shape child development, many developmental analyses have focused on analyzing cultural conceptions of independence and interdependence. Less attention has been paid to charting the developmental sequences of children's independent and interdependent behavior that are ostensibly shaped by cultural…

  13. To Encounter, to Build the World and to Become a Human Being. Advocating for a Material-Cultural Turn in Developmental Psychology.

    PubMed

    Moro, Christiane

    2016-12-01

    Why have material world of daily life and material objects in their conventional features or to say it in other words, why have the mundane world and mundane objects, in which the human beings live and children come to, encounter, experience and develop through, received so little attention from psychologists thus remaining a blind spot in mainstream developmental psychology? Certainly the object has not been totally forgotten (e.g. Piaget's constructivist paradigm) but it has been considered as theoretically determined by the categories of understanding (cf. Kant), and considered as a key to understanding the world in its physical properties by the infant. But the material world and the material objects that are used for everyday purposes (i.e. pragmatically) belonging to material culture, have been totally neglected by developmental psychologists. Reacting to the Kantian agenda of developmental psychology but also to heterodox non developmentalist thinkers such as Gibson who is a growing source of inspiration for developmental psychologists today, we challenge the taken-for-granted mundane world, arguing for the importance of material objects related to material culture in psychological development during the prelinguistic period. On the basis of recent research in early development grounded in the Vygotskian paradigm, we discuss this issue through Marxist Anthropology, Material Culture Studies and Phenomenology. As a consequence we advocate for a material-cultural turn in psychological development in order to place the issue of material world and material objects in their pragmatic and semiotic features on the agenda of developmental psychology.

  14. Cultural variations in global versus local processing: a developmental perspective.

    PubMed

    Oishi, Shigehiro; Jaswal, Vikram K; Lillard, Angeline S; Mizokawa, Ai; Hitokoto, Hidefumi; Tsutsui, Yoshiro

    2014-12-01

    We conducted 3 studies to explore cultural differences in global versus local processing and their developmental trajectories. In Study 1 (N = 363), we found that Japanese college students were less globally oriented in their processing than American or Argentine participants. We replicated this effect in Study 2 (N = 1,843) using a nationally representative sample of Japanese and American adults ages 20 to 69, and found further that adults in both cultures became more globally oriented with age. In Study 3 (N = 133), we investigated the developmental course of the cultural difference using Japanese and American children, and found it was evident by 4 years of age. Cultural variations in global versus local processing emerge by early childhood, and remain throughout adulthood. At the same time, both Japanese and Americans become increasingly global processors with age. PsycINFO Database Record (c) 2014 APA, all rights reserved.

  15. Efficient embryonic culture method for the Japanese striped snake, Elaphe quadrivirgata, and its early developmental stages.

    PubMed

    Matsubara, Yoshiyuki; Sakai, Atsushi; Kuroiwa, Atsushi; Suzuki, Takayuki

    2014-10-01

    The morphogenesis of snake embryos is an elusive yet fascinating research target for developmental biologists. However, few data exist on development of early snake embryo due to limited availability of pregnant snakes, and the need to harvest early stage embryos directly from pregnant snakes before oviposition without knowing the date of fertilization. We established an ex vivo culture method for early snake embryos using the Japanese striped snake, Elaphe quadrivirgata. This method, which we named "sausage-style (SS) culture", allows us to harvest snake embryos at specific stages for each experiment. Using this SS culture system, we calculated somite formation rate at early stages before oviposition. The average somite formation rate between 6/7 and 12/13 somite stages was 145.9 min, between 60/70 and 80/91 somite stages 42.4 min, and between 113-115 and 126/127 somite stages 71 min. Thus, somite formation rate that we observed during early snake embryogenesis was changed over time. We also describe a developmental staging series for E. quadrivirgata. This is the first report of a developmental series of early snake embryogenesis prior to oviposition by full-color images with high-resolution. We propose that the SS culture system is an easy method for treating early snake embryos ex vivo. © 2014 The Authors Development, Growth & Differentiation © 2014 Japanese Society of Developmental Biologists.

  16. Basic procedures for epigenetic analysis in plant cell and tissue culture.

    PubMed

    Rodríguez, José L; Pascual, Jesús; Viejo, Marcos; Valledor, Luis; Meijón, Mónica; Hasbún, Rodrigo; Yrei, Norma Yague; Santamaría, María E; Pérez, Marta; Fernández Fraga, Mario; Berdasco, María; Rodríguez Fernández, Roberto; Cañal, María J

    2012-01-01

    In vitro culture is one of the most studied techniques, and it is used to study many developmental processes, especially in forestry species, because of growth timing and easy manipulation. Epigenetics has been shown as an important influence on many research analyses such as cancer in mammals and developmental processes in plants such as flowering, but regarding in vitro culture, techniques to study DNA methylation or chromatin modifications were mainly limited to identify somaclonal variation of the micropropagated material. Because in vitro culture is not only a way to generate plant material but also a bunch of differentially induced developmental processes, an approach of techniques and some research carried out to study the different changes regarding DNA methylation and chromatin and translational modifications that take place during these processes is reviewed.

  17. Beyond Borders: Poetry Slicing through Steel Gates and Barbed Wires

    ERIC Educational Resources Information Center

    Jocson, Korina M.

    2004-01-01

    Exchange of poems at the 2nd Annual San Quentin/Patten College poetry slam with the prisoners is reported to be an event, which was extraordinaire. It was an opportunity to understand the hidden popular culture.

  18. Slicing for Biology.

    ERIC Educational Resources Information Center

    Ekstrom, James

    2001-01-01

    Advocates using computer imaging technology to assist students in doing projects in which determining density is important. Students can study quantitative comparisons of masses, lengths, and widths using computer software. Includes figures displaying computer images of shells, yeast cultures, and the Aral Sea. (SAH)

  19. "Ganando Confianza": Research Focus Groups with Immigrant Mexican Mothers

    ERIC Educational Resources Information Center

    Hausmann-Stabile, Carolina; Zayas, Luis H.; Runes, Sandra; Abenis-Cintron, Anna; Calzada, Esther

    2011-01-01

    Immigrant families with children with developmental disabilities must be served using culturally sensitive approaches to service and research to maximize treatment benefits. In an effort to better understand cultural issues relevant to the provision of parenting programs for immigrant Mexican mothers of children with developmental disabilities, we…

  20. In Search of Theory: The Study of "Ethnic Groups" in Developmental Psychology.

    ERIC Educational Resources Information Center

    Gjerde, Per F.; Onishi, Miyoko

    2000-01-01

    Discusses the conceptual status and uses of ethnic groups in developmental psychology. Discusses problems with the primordialist position and the influence of nationalism in defining culture. Argues that culture and ethnicity as shared and located within a bounded population is an increasingly outmoded notion. Maintains that developmental…

  1. Cultural Variations in Global versus Local Processing: A Developmental Perspective

    ERIC Educational Resources Information Center

    Oishi, Shigehiro; Jaswal, Vikram K.; Lillard, Angeline S.; Mizokawa, Ai; Hitokoto, Hidefumi; Tsutsui, Yoshiro

    2014-01-01

    We conducted 3 studies to explore cultural differences in global versus local processing and their developmental trajectories. In Study 1 ("N" = 363), we found that Japanese college students were less globally oriented in their processing than American or Argentine participants. We replicated this effect in Study 2 ("N" =…

  2. The (Be)Comings and Goings of "Developmental Disabilities": The Cultural Politics of "Impairment"

    ERIC Educational Resources Information Center

    Goodley, Dan; Roets, Griet

    2008-01-01

    Disability should be a concern for those interested in analysing and subverting the cultural politics of education. In this paper we address this concern through connecting critical analyses of "developmental disabilities" (formerly "mental retardation"), disability studies and poststructuralism. We target normative constructions of "developmental…

  3. Targeting neurotransmitter receptors with nanoparticles in vivo allows single-molecule tracking in acute brain slices

    NASA Astrophysics Data System (ADS)

    Varela, Juan A.; Dupuis, Julien P.; Etchepare, Laetitia; Espana, Agnès; Cognet, Laurent; Groc, Laurent

    2016-03-01

    Single-molecule imaging has changed the way we understand many biological mechanisms, particularly in neurobiology, by shedding light on intricate molecular events down to the nanoscale. However, current single-molecule studies in neuroscience have been limited to cultured neurons or organotypic slices, leaving as an open question the existence of fast receptor diffusion in intact brain tissue. Here, for the first time, we targeted dopamine receptors in vivo with functionalized quantum dots and were able to perform single-molecule tracking in acute rat brain slices. We propose a novel delocalized and non-inflammatory way of delivering nanoparticles (NPs) in vivo to the brain, which allowed us to label and track genetically engineered surface dopamine receptors in neocortical neurons, revealing inherent behaviour and receptor activity regulations. We thus propose a NP-based platform for single-molecule studies in the living brain, opening new avenues of research in physiological and pathological animal models.

  4. Variation is the universal: making cultural evolution work in developmental psychology.

    PubMed

    Kline, Michelle Ann; Shamsudheen, Rubeena; Broesch, Tanya

    2018-04-05

    Culture is a human universal, yet it is a source of variation in human psychology, behaviour and development. Developmental researchers are now expanding the geographical scope of research to include populations beyond relatively wealthy Western communities. However, culture and context still play a secondary role in the theoretical grounding of developmental psychology research, far too often. In this paper, we highlight four false assumptions that are common in psychology, and that detract from the quality of both standard and cross-cultural research in development. These assumptions are: (i) the universality assumption , that empirical uniformity is evidence for universality, while any variation is evidence for culturally derived variation; (ii) the Western centrality assumption , that Western populations represent a normal and/or healthy standard against which development in all societies can be compared; (iii) the deficit assumption , that population-level differences in developmental timing or outcomes are necessarily due to something lacking among non-Western populations; and (iv) the equivalency assumption , that using identical research methods will necessarily produce equivalent and externally valid data, across disparate cultural contexts. For each assumption, we draw on cultural evolutionary theory to critique and replace the assumption with a theoretically grounded approach to culture in development. We support these suggestions with positive examples drawn from research in development. Finally, we conclude with a call for researchers to take reasonable steps towards more fully incorporating culture and context into studies of development, by expanding their participant pools in strategic ways. This will lead to a more inclusive and therefore more accurate description of human development.This article is part of the theme issue 'Bridging cultural gaps: interdisciplinary studies in human cultural evolution'. © 2018 The Author(s).

  5. Localizing Protein in 3D Neural Stem Cell Culture: a Hybrid Visualization Methodology

    PubMed Central

    Fai, Stephen; Bennett, Steffany A.L.

    2010-01-01

    The importance of 3-dimensional (3D) topography in influencing neural stem and progenitor cell (NPC) phenotype is widely acknowledged yet challenging to study. When dissociated from embryonic or post-natal brain, single NPCs will proliferate in suspension to form neurospheres. Daughter cells within these cultures spontaneously adopt distinct developmental lineages (neurons, oligodendrocytes, and astrocytes) over the course of expansion despite being exposed to the same extracellular milieu. This progression recapitulates many of the stages observed over the course of neurogenesis and gliogenesis in post-natal brain and is often used to study basic NPC biology within a controlled environment. Assessing the full impact of 3D topography and cellular positioning within these cultures on NPC fate is, however, difficult. To localize target proteins and identify NPC lineages by immunocytochemistry, free-floating neurospheres must be plated on a substrate or serially sectioned. This processing is required to ensure equivalent cell permeabilization and antibody access throughout the sphere. As a result, 2D epifluorescent images of cryosections or confocal reconstructions of 3D Z-stacks can only provide spatial information about cell position within discrete physical or digital 3D slices and do not visualize cellular position in the intact sphere. Here, to reiterate the topography of the neurosphere culture and permit spatial analysis of protein expression throughout the entire culture, we present a protocol for isolation, expansion, and serial sectioning of post-natal hippocampal neurospheres suitable for epifluorescent or confocal immunodetection of target proteins. Connexin29 (Cx29) is analyzed as an example. Next, using a hybrid of graphic editing and 3D modelling softwares rigorously applied to maintain biological detail, we describe how to re-assemble the 3D structural positioning of these images and digitally map labelled cells within the complete neurosphere. This methodology enables visualization and analysis of the cellular position of target proteins and cells throughout the entire 3D culture topography and will facilitate a more detailed analysis of the spatial relationships between cells over the course of neurogenesis and gliogenesis in vitro. Both Imbeault and Valenzuela contributed equally and should be considered joint first authors. PMID:21258319

  6. An organotypic slice culture model of chronic white matter injury with maturation arrest of oligodendrocyte progenitors.

    PubMed

    Dean, Justin M; Riddle, Art; Maire, Jennifer; Hansen, Kelly D; Preston, Marnie; Barnes, Anthony P; Sherman, Larry S; Back, Stephen A

    2011-07-05

    CNS myelination disturbances commonly occur in chronic white matter lesions in neurodevelopmental and adult neurological disorders. Recent studies support that myelination failure can involve a disrupted cellular repair mechanism where oligodendrocyte (OL) progenitor cells (OPCs) proliferate in lesions with diffuse astrogliosis, but fail to fully differentiate to mature myelinating OLs. There are no in vitro models that reproduce these features of myelination failure. Forebrain coronal slices from postnatal day (P) 0.5/1 rat pups were cultured for 1, 5, or 9 days in vitro (DIV). Slices rapidly exhibited diffuse astrogliosis and accumulation of the extracellular matrix glycosaminoglycan hyaluronan (HA), an inhibitor of OPC differentiation and re-myelination. At 1 DIV ~1.5% of Olig2+ OLs displayed caspase-3 activation, which increased to ~11.5% by 9 DIV. At 1 DIV the density of PDGFRα+ and PDGFRα+/Ki67+ OPCs were significantly elevated compared to 0 DIV (P < 0.01). Despite this proliferative response, at 9 DIV ~60% of white matter OLs were late progenitors (preOLs), compared to ~7% in the postnatal day 10 rat (P < 0.0001), consistent with preOL maturation arrest. Addition of HA to slices significantly decreased the density of MBP+ OLs at 9 DIV compared to controls (217 ± 16 vs. 328 ± 17 cells/mm2, respectively; P = 0.0003), supporting an inhibitory role of HA in OL lineage progression in chronic lesions. Diffuse white matter astrogliosis and early OPC proliferation with impaired OL maturation were reproduced in this model of myelination failure. This system may be used to define mechanisms of OPC maturation arrest and myelination failure related to astrogliosis and HA accumulation.

  7. Beyond Race: Examining the Facets of Multiracial Identity through a Life-Span Developmental Lens

    ERIC Educational Resources Information Center

    Jackson, Kelly F.

    2009-01-01

    Using a social work developmental lens, this qualitative study explored some of the numerous social and environmental factors that shape a multiracial individual's cultural identity. Results from transcript analysis portray the cultural identity of multiracial persons as significantly influenced by (1) personal experiences of racism and…

  8. Fostering Culturally and Developmentally Responsive Teaching through Improvisational Practice

    ERIC Educational Resources Information Center

    Graue, Elizabeth; Whyte, Kristin; Delaney, Kate Kresin

    2014-01-01

    In this article we explore an effort to rethink curricular decision-making with a group of public pre-K teachers working in a context of curriculum escalation and commitment to play-based pedagogy. Through a professional development program designed to support developmentally and culturally responsive early mathematics, we examine how teachers…

  9. Mexican American Adolescent Couples Communicating about Conflict: An Integrated Developmental and Cultural Perspective

    ERIC Educational Resources Information Center

    Rueda, Heidi Adams; Williams, Lela Rankin

    2016-01-01

    Using observational methods on a small sample of committed Mexican American couples (N = 10, ages 15-17, M length of relationship = 26.5 months), we describe and categorize developmental and cultural communication patterns concerning the negotiation of conflict issues. Videotaped dyadic interactions were transcribed and qualitatively coded using…

  10. Culture and Developmental Trajectories: A Discussion on Contemporary Theoretical Models

    ERIC Educational Resources Information Center

    de Carvalho, Rafael Vera Cruz; Seidl-de-Moura, Maria Lucia; Martins, Gabriela Dal Forno; Vieira, Mauro Luís

    2014-01-01

    This paper aims to describe, compare and discuss the theoretical models proposed by Patricia Greenfield, Çigdem Kagitçibasi and Heidi Keller. Their models have the common goal of understanding the developmental trajectories of self based on dimensions of autonomy and relatedness that are structured according to specific cultural and environmental…

  11. Developmental roots of episodic memory.

    PubMed

    Nelson, Katherine

    2018-01-01

    Two arguments imply that Mahr & Csibra's (M&C's) functional theory is insufficient as an explanation of episodic memory: (1) The developmental course supports a different social cultural division of episodic and semantic memory, and (2) the existence of long-term autobiographical memory is not explained in the functional theory but can be seen in a broader cultural framework.

  12. Cross-Cultural Adaptation of Developmental Criteria for Young Children: A Preliminary Psychometric Study

    ERIC Educational Resources Information Center

    Yunus, Faridah

    2014-01-01

    Authentic assessment approach applies naturalistic observation method to gather and analyse data about children's development that are socio-culturally appropriate to plan for individual teaching and learning needs. This article discusses the process of adapting an authentic developmental instrument for children of 3-6 years old. The instrument…

  13. NCT and Developmental Psychology: A Welcome Rapprochement

    ERIC Educational Resources Information Center

    Gauvain, Mary

    2013-01-01

    For over 50 years, developmental psychologists have conducted research around the world to understand the relation between culture and cognition. In fact, psychologists have been interested in this topic for over a century. In the late 1800s, Wundt introduced "Elements of Folk Psychology," the study of how culture becomes part of higher…

  14. Team Creativity: The Effects of Perceived Learning Culture, Developmental Feedback and Team Cohesion

    ERIC Educational Resources Information Center

    Joo, Baek-Kyoo; Song, Ji Hoon; Lim, Doo Hun; Yoon, Seung Won

    2012-01-01

    This study investigates the influence of perceived learning culture, developmental feedback and team cohesion on team creativity. The results showed that the demographic variables, the three antecedents and their interactions explained 41 per cent of variance in team creativity. Team creativity was positively correlated with a higher level of…

  15. Brain Slice Staining and Preparation for Three-Dimensional Super-Resolution Microscopy

    PubMed Central

    German, Christopher L.; Gudheti, Manasa V.; Fleckenstein, Annette E.; Jorgensen, Erik M.

    2018-01-01

    Localization microscopy techniques – such as photoactivation localization microscopy (PALM), fluorescent PALM (FPALM), ground state depletion (GSD), and stochastic optical reconstruction microscopy (STORM) – provide the highest precision for single molecule localization currently available. However, localization microscopy has been largely limited to cell cultures due to the difficulties that arise in imaging thicker tissue sections. Sample fixation and antibody staining, background fluorescence, fluorophore photoinstability, light scattering in thick sections, and sample movement create significant challenges for imaging intact tissue. We have developed a sample preparation and image acquisition protocol to address these challenges in rat brain slices. The sample preparation combined multiple fixation steps, saponin permeabilization, and tissue clarification. Together, these preserve intracellular structures, promote antibody penetration, reduce background fluorescence and light scattering, and allow acquisition of images deep in a 30 μm thick slice. Image acquisition challenges were resolved by overlaying samples with a permeable agarose pad and custom-built stainless steel imaging adapter, and sealing the imaging chamber. This approach kept slices flat, immobile, bathed in imaging buffer, and prevented buffer oxidation during imaging. Using this protocol, we consistently obtained single molecule localizations of synaptic vesicle and active zone proteins in three-dimensions within individual synaptic terminals of the striatum in rat brain slices. These techniques may be easily adapted to the preparation and imaging of other tissues, substantially broadening the application of super-resolution imaging. PMID:28924666

  16. Linking social change and developmental change: shifting pathways of human development.

    PubMed

    Greenfield, Patricia M

    2009-03-01

    P. M. Greenfield's new theory of social change and human development aims to show how changing sociodemographic ecologies alter cultural values and learning environments and thereby shift developmental pathways. Worldwide sociodemographic trends include movement from rural residence, informal education at home, subsistence economy, and low-technology environments to urban residence, formal schooling, commerce, and high-technology environments. The former ecology is summarized by the German term Gemeinschaft ("community") and the latter by the German term Gesellschaft ("society"; Tönnies, 1887/1957). A review of empirical research demonstrates that, through adaptive processes, movement of any ecological variable in a Gesellschaft direction shifts cultural values in an individualistic direction and developmental pathways toward more independent social behavior and more abstract cognition--to give a few examples of the myriad behaviors that respond to these sociodemographic changes. In contrast, the (much less frequent) movement of any ecological variable in a Gemeinschaft direction is predicted to move cultural values and developmental pathways in the opposite direction. In conclusion, sociocultural environments are not static either in the developed or the developing world and therefore must be treated dynamically in developmental research.

  17. Antibacterial effects of Nd:YAG laser irradiation within root canal dentin.

    PubMed

    Klinke, T; Klimm, W; Gutknecht, N

    1997-02-01

    The microbial flora of the root canal dentin can cause failures in the conventional treatment of infected root canals if it cannot be sufficiently removed by preparation and chemical disinfection of the root canal. The aim of this study is to examine the bactericidal effects of neodymium:yttriumaluminum garnet (Nd:YAG) laser irradiation in the depth of the root canal dentin. Following sterilization, longitudinal section dentin slices of different thicknesses (100-1000 microns) were inoculated on one side with 4 microliters of a Streptococcus mutans suspension. The opposite sides of the dentin slices were then irradiated four times for 10-20 sec (according to the sample area) using the Nd:YAG laser at a setting of 1.5 W, 15 pps with a 200 microns glass fiber from an angle of about 5 degrees. The bacteria were then removed from the dentin using vibration and plated out on culture dishes that were selective for Streptococcus mutans. When compared with untreated control slices, counting of the colonies revealed a highly significant elimination of bacteria for all thicknesses following laser irradiation. Although the intensity of the laser irradiation decreased after penetration of a 1000-micron dentin slice, the bactericidal mode of action was still effective.

  18. A cross-cultural investigation of inhibitory control, generative fluency, and anxiety symptoms in Romanian and Russian preschoolers.

    PubMed

    Cheie, Lavinia; Veraksa, Aleksander; Zinchenko, Yuri; Gorovaya, Alexandra; Visu-Petra, Laura

    2015-01-01

    The current study focused on the early development of inhibitory control in 5- to 7-year-old children attending kindergarten in two Eastern-European countries, Romania and Russia. These two countries share many aspects of child-rearing and educational practices, previously documented to influence the development of inhibitory control. Using the Lurian-based developmental approach offered by the Developmental Neuropsychological Assessment battery, the study aimed to contribute to cross-cultural developmental neuropsychology by exploring (a) early interrelationships between subcomponents of inhibitory control (response suppression and attention control) and generative fluency (verbal and figural) in these two cultures, as well as (b) the predictive value of external factors (culture and maternal education) and individual differences (age, gender, nonverbal intelligence, trait anxiety) on inhibitory control and fluency outcomes in children from both countries. First, findings in both culture samples suggest that even at this young age, the construct of inhibitory control cannot be considered a unitary entity. Second, differences in maternal education were not predictive of either inhibitory control or fluency scores. However, children's attention control performance varied as a function of culture, and the direction of these cultural effects differed by whether the target outcome involved performance accuracy versus efficiency as an output. Findings also confirmed the previously documented intensive developmental improvement in preschoolers' inhibitory control during this period, influencing measures of response suppression and particularly attention control. Finally, the results further stress the importance of individual differences effects in trait anxiety on attention control efficiency across cultures.

  19. Focal macromolecule delivery in neuronal tissue using simultaneous pressure ejection and local electroporation

    PubMed Central

    Barker, Matthew; Billups, Brian; Hamann, Martine

    2009-01-01

    Electroporation creates transient pores in the plasma membrane to introduce macromolecules within a cell or cell population. Generally, electrical pulses are delivered between two electrodes separated from each other, making electroporation less likely to be localised. We have developed a new device combining local pressure ejection with local electroporation through a double-barrelled glass micropipette to transfer impermeable macromolecules in brain slices or in cultured HEK293 cells. The design achieves better targeting of the site of pressure ejection with that of electroporation. With this technique, we have been able to limit the delivery of propidium iodide or dextran amine within areas of 100–200 μm diameter. We confirm that local electroporation is transient and show that when combined with pressure ejection, it allows local transfection of EGFP plasmids within HEK293 cells or within cerebellar and hippocampal slice cultures. We further show that local electroporation is less damaging when compared to global electroporation using two separate electrodes. Focal delivery of dextran amine dyes within trapezoid body fibres allowed tracing axonal tracts within brainstem slices, enabling the study of identified calyx of Held presynaptic terminals in living brain tissue. This labelling method can be used to target small nuclei in neuronal tissue and is generally applicable to the study of functional synaptic connectivity, or live axonal tracing in a variety of brain areas. PMID:19014970

  20. Agonist-induced glycogenolysis in rabbit retinal slices and cultures.

    PubMed Central

    Ghazi, H.; Osborne, N. N.

    1989-01-01

    1. The effects of different putative retinal transmitters and/or modulators on glycogenolysis in rabbit retinal slices and in retinal Müller cell cultures were examined. 2. Incubation of rabbit retinal slices or primary retinal cultures (either 3-5 day-old or 25-30 day-old) in a buffer solution containing [3H]-glucose resulted in the accumulation of newly synthesized [3H]-glycogen. 3. Noradrenaline (NA), isoprenaline, vasoactive intestinal peptide (VIP), 5-hydroxytryptamine (5-HT) and 8-hydroxy-dipropylaminetetralin (8-OH-DPAT) stimulated the hydrolysis of this newly formed 3H-polymer. The potency order of maximal stimulations was: VIP greater than NA greater than isoprenaline greater than 5-HT greater than 8-OH-DPAT. 4. The putative retinal transmitters, dopamine, gamma-aminobutyric acid (GABA), glycine and taurine and the muscarinic agonist carbachol (CCh) had no effect on [3H]-glycogen content. 5. The glycogenolytic effects of NA/isoprenaline and 5-HT/8-OH-DPAT appear to be mediated by beta-adrenoceptors and 5-HT1 receptors (possibly 5-HT1A), respectively while the VIP-induced response involved another receptor subtype. 6. Agonists which mediated [3H]-glycogen hydrolysis also stimulated an increase in adenosine 3':5'-cyclic monophosphate (cyclic AMP) formation. Both responses are blocked to a similar extent by the same antagonists and so are probably mediated via the same receptor subtypes. Moreover, dibutyryl cyclic AMP (db cyclic AMP) promoted tritiated glycogen breakdown in the three retinal preparations. 7. Not all receptors linked to cyclic AMP production however promote glycogenolysis. Dopamine and apomorphine stimulated cyclic AMP formation via D1-receptors without influencing glycogenolysis. These receptors are exclusively associated with neurones. PMID:2568145

  1. Blood cell oxidative stress precedes hemolysis in whole blood-liver slice co-cultures of rat, dog, and human tissues

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

    Vickers, Alison E.M., E-mail: vickers_alison@allergan.co; Sinclair, John R.; Fisher, Robyn L.

    A novel in vitro model to investigate time-dependent and concentration-dependent responses in blood cells and hemolytic events is studied for rat, dog, and human tissues. Whole blood is co-cultured with a precision-cut liver slice. Methimazole (MMI) was selected as a reference compound, since metabolism of its imidazole thione moiety is linked with hematologic disorders and hepatotoxicity. An oxidative stress response occurred in all three species, marked by a decline in blood GSH levels by 24 h that progressed, and preceded hemolysis, which occurred at high MMI concentrations in the presence of a liver slice with rat (>= 1000 muM atmore » 48 h) and human tissues (>= 1000 muM at 48 h, >= 750 muM at 72 h) but not dog. Human blood-only cultures exhibited a decline of GSH levels but minimal to no hemolysis. The up-regulation of liver genes for heme degradation (Hmox1 and Prdx1), iron cellular transport (Slc40a1), and GSH synthesis and utilization (mGST1 and Gclc) were early markers of the oxidative stress response. The up-regulation of the Kupffer cell lectin Lgals3 gene expression indicated a response to damaged red blood cells, and Hp (haptoglobin) up-regulation is indicative of increased hemoglobin uptake. Up-regulation of liver IL-6 and IL-8 gene expression suggested an activation of an inflammatory response by liver endothelial cells. In summary, MMI exposure led to an oxidative stress response in blood cells, and an up-regulation of liver genes involved with oxidative stress and heme homeostasis, which was clearly separate and preceded frank hemolysis.« less

  2. Chronic pharmacological blockade of the Na+ /Ca2+ exchanger modulates the growth and development of the Purkinje cell dendritic arbor in mouse cerebellar slice cultures.

    PubMed

    Sherkhane, Pradeep; Kapfhammer, Josef P

    2017-09-01

    The Na + /Ca 2+ exchanger (NCX) is a bidirectional plasma membrane antiporter involved in Ca 2+ homeostasis in eukaryotes. NCX has three isoforms, NCX1-3, and all of them are expressed in the cerebellum. Immunostaining on cerebellar slice cultures indicates that NCX is widely expressed in the cerebellum, including expression in Purkinje cells. The pharmacological blockade of the forward mode of NCX (Ca 2+ efflux mode) by bepridil moderately inhibited growth and development of Purkinje cell dendritic arbor in cerebellar slice cultures. However, the blockade of the reverse mode (Ca 2+ influx mode) by KB-R7943 severely reduced the dendritic arbor and induced a morphological change with thickened distal dendrites. The effect of KB-R7943 on dendritic growth was unrelated to the activity of voltage-gated calcium channels and was also apparent in the absence of bioelectrical activity indicating that it was mediated by NCX expressed in Purkinje cells. We have used additional NCX inhibitors including CB-DMB, ORM-10103, SEA0400, YM-244769, and SN-6 which have higher specificity for NCX isoforms and target either the forward, reverse, or both modes. These inhibitors caused a strong dendritic reduction similar to that seen with KB-R7943, but did not elicit thickening of distal dendrites. Our findings indicate that disturbance of the NCX-dependent calcium transport in Purkinje cells induces a reduction of dendritic arbor, which is presumably caused by changes in the calcium handling, and underline the importance of the calcium equilibrium for the dendritic development in cerebellar Purkinje cells. © 2017 Federation of European Neuroscience Societies and John Wiley & Sons Ltd.

  3. Hyperpolarized 13C MR Markers of Renal Tumor Aggressiveness

    DTIC Science & Technology

    2013-10-01

    slice cultures , bioreactor 16. SECURITY CLASSIFICATION OF: 17. LIMITATION OF ABSTRACT 18. NUMBER OF PAGES 19a. NAME OF RESPONSIBLE PERSON...tumors from RCCs, and low grade from high grade RCCs using human TSCs cultured in a bioreactor . Aim 2:Identify HP 13C metabolic markers that...and cells over 48 hours using fluorescent dyes incubated with TSCs 1 Aim 1: Ex vivo bioreactor experiments We first acquired phosphorous-31 (31P

  4. Dimensions of Conflict for Youth: A Cross-Cultural View. Seminar Series No. 5.

    ERIC Educational Resources Information Center

    Belbas, Nancy, Ed.

    This document presents, in two parts, ideas and information which were gathered from eight monthly seminars. Part 1, "Developmental and Cross-Cultural Patterns of Youth," examines what is known about developmental norms regarding youth in general in respect to their attitudes at different ages towards rules and authority. It is found that in the…

  5. MPX-004 and MPX-007: New Pharmacological Tools to Study the Physiology of NMDA Receptors Containing the GluN2A Subunit.

    PubMed

    Volkmann, Robert A; Fanger, Christopher M; Anderson, David R; Sirivolu, Venkata Ramana; Paschetto, Kathy; Gordon, Earl; Virginio, Caterina; Gleyzes, Melanie; Buisson, Bruno; Steidl, Esther; Mierau, Susanna B; Fagiolini, Michela; Menniti, Frank S

    2016-01-01

    GluN2A is the most abundant of the GluN2 NMDA receptor subunits in the mammalian CNS. Physiological and genetic evidence implicate GluN2A-containing receptors in susceptibility to autism, schizophrenia, childhood epilepsy and neurodevelopmental disorders such as Rett Syndrome. However, GluN2A-selective pharmacological probes to explore the therapeutic potential of targeting these receptors have been lacking. Here we disclose a novel series of pyrazine-containing GluN2A antagonists exemplified by MPX-004 (5-(((3-chloro-4-fluorophenyl)sulfonamido)methyl)-N-((2-methylthiazol-5-yl)methyl)pyrazine-2-carboxamide) and MPX-007 (5-(((3-fluoro-4-fluorophenyl)sulfonamido)methyl)-N-((2-methylthiazol-5-yl)methyl)methylpyrazine-2-carboxamide). MPX-004 and MPX-007 inhibit GluN2A-containing NMDA receptors expressed in HEK cells with IC50s of 79 nM and 27 nM, respectively. In contrast, at concentrations that completely inhibited GluN2A activity these compounds have no inhibitory effect on GluN2B or GluN2D receptor-mediated responses in similar HEK cell-based assays. Potency and selectivity were confirmed in electrophysiology assays in Xenopus oocytes expressing GluN2A-D receptor subtypes. Maximal concentrations of MPX-004 and MPX-007 inhibited ~30% of the whole-cell current in rat pyramidal neurons in primary culture and MPX-004 inhibited ~60% of the total NMDA receptor-mediated EPSP in rat hippocampal slices. GluN2A-selectivity at native receptors was confirmed by the finding that MPX-004 had no inhibitory effect on NMDA receptor mediated synaptic currents in cortical slices from GRIN2A knock out mice. Thus, MPX-004 and MPX-007 offer highly selective pharmacological tools to probe GluN2A physiology and involvement in neuropsychiatric and developmental disorders.

  6. The Electrophysiological MEMS Device with Micro Channel Array for Cellular Network Analysis

    NASA Astrophysics Data System (ADS)

    Tonomura, Wataru; Kurashima, Toshiaki; Takayama, Yuzo; Moriguchi, Hiroyuki; Jimbo, Yasuhiko; Konishi, Satoshi

    This paper describes a new type of MCA (Micro Channel Array) for simultaneous multipoint measurement of cellular network. Presented MCA employing the measurement principles of the patch-clamp technique is designed for advanced neural network analysis which has been studied by co-authors using 64ch MEA (Micro Electrode Arrays) system. First of all, sucking and clamping of cells through channels of developed MCA is expected to improve electrophysiological signal detections. Electrophysiological sensing electrodes integrated around individual channels of MCA by using MEMS (Micro Electro Mechanical System) technologies are electrically isolated for simultaneous multipoint measurement. In this study, we tested the developed MCA using the non-cultured rat's cerebral cortical slice and the hippocampal neurons. We could measure the spontaneous action potential of the slice simultaneously at multiple points and culture the neurons on developed MCA. Herein, we describe the experimental results together with the design and fabrication of the electrophysiological MEMS device with MCA for cellular network analysis.

  7. Fast two-photon imaging of subcellular voltage dynamics in neuronal tissue with genetically encoded indicators.

    PubMed

    Chamberland, Simon; Yang, Helen H; Pan, Michael M; Evans, Stephen W; Guan, Sihui; Chavarha, Mariya; Yang, Ying; Salesse, Charleen; Wu, Haodi; Wu, Joseph C; Clandinin, Thomas R; Toth, Katalin; Lin, Michael Z; St-Pierre, François

    2017-07-27

    Monitoring voltage dynamics in defined neurons deep in the brain is critical for unraveling the function of neuronal circuits but is challenging due to the limited performance of existing tools. In particular, while genetically encoded voltage indicators have shown promise for optical detection of voltage transients, many indicators exhibit low sensitivity when imaged under two-photon illumination. Previous studies thus fell short of visualizing voltage dynamics in individual neurons in single trials. Here, we report ASAP2s, a novel voltage indicator with improved sensitivity. By imaging ASAP2s using random-access multi-photon microscopy, we demonstrate robust single-trial detection of action potentials in organotypic slice cultures. We also show that ASAP2s enables two-photon imaging of graded potentials in organotypic slice cultures and in Drosophila . These results demonstrate that the combination of ASAP2s and fast two-photon imaging methods enables detection of neural electrical activity with subcellular spatial resolution and millisecond-timescale precision.

  8. Electroosmotic perfusion of tissue: sampling the extracellular space and quantitative assessment of membrane-bound enzyme activity in organotypic hippocampal slice cultures.

    PubMed

    Ou, Yangguang; Wu, Juanfang; Sandberg, Mats; Weber, Stephen G

    2014-10-01

    This review covers recent advances in sampling fluid from the extracellular space of brain tissue by electroosmosis (EO). Two techniques, EO sampling with a single fused-silica capillary and EO push-pull perfusion, have been developed. These tools were used to investigate the function of membrane-bound enzymes with outward-facing active sites, or ectoenzymes, in modulating the activity of the neuropeptides leu-enkephalin and galanin in organotypic-hippocampal-slice cultures (OHSCs). In addition, the approach was used to determine the endogenous concentration of a thiol, cysteamine, in OHSCs. We have also investigated the degradation of coenzyme A in the extracellular space. The approach provides information on ectoenzyme activity, including Michaelis constants, in tissue, which, as far as we are aware, has not been done before. On the basis of computational evidence, EO push-pull perfusion can distinguish ectoenzyme activity with a ~100 μm spatial resolution, which is important for studies of enzyme kinetics in adjacent regions of the rat hippocampus.

  9. Ketone bodies do not directly alter excitatory or inhibitory hippocampal synaptic transmission.

    PubMed

    Thio, L L; Wong, M; Yamada, K A

    2000-01-25

    To determine the effect of the ketone bodies beta-hydroxybutyrate (betaHB) and acetoacetate (AA) on excitatory and inhibitory neurotransmission in the mammalian CNS. The ketogenic diet is presumed to be an effective anticonvulsant regimen for some children with medically intractable seizures. However, its mechanism of action remains a mystery. According to one hypothesis, ketone bodies have anticonvulsant properties. The authors examined the effect of betaHB and AA on excitatory and inhibitory synaptic transmission in rat hippocampal-entorhinal cortex slices and cultured hippocampal neurons. In cultured neurons, their effect was also directly assayed on postsynaptic receptor properties. Finally, their ability to prevent spontaneous seizures was determined in a hippocampal-entorhinal cortex slice model. betaHB and AA did not alter synaptic transmission in these models. The anticonvulsant properties of the ketogenic diet do not result from a direct effect of ketone bodies on the primary voltage and ligand gated ion channels mediating excitatory or inhibitory neurotransmission in the hippocampus.

  10. Developing culturally responsive approaches with Southeast Asian American families experiencing developmental disabilities.

    PubMed

    Baker, Dian L; Miller, Elizabeth; Dang, Michelle T; Yaangh, Chiem-Seng; Hansen, Robin L

    2010-12-01

    Southeast Asian American families are underrepresented among recipients of special education and social services for people with developmental disabilities. Our aims were to use a community-based participatory research approach to examine Hmong and Mien families' perceptions of developmental disabilities and understand barriers to and facilitators of service provision among families experiencing developmental disabilities. We describe here a case study of a successful attempt to engage marginalized and underserved communities to understand their needs to improve access and services for persons with developmental disabilities. We conducted 2 focus groups with 11 key informants and 1 focus group with 10 family members of persons with developmental disabilities, as well as in-depth interviews with 3 shamans. Using a thematic analysis approach, we coded notes and transcripts to assess community members' understanding of developmental disabilities, experiences negotiating educational and health care systems, and barriers to high-quality care. A predominant theme was the perception that reliance on governmental support services is not appropriate. Common barriers identified included lack of accurate information, language difficulties, lack of trust, and limited outreach. These perceptions and barriers, combined with limited access to services, interfere with community acceptance and use of available support services. Despite these barriers, participants indicated that with education, outreach, and culturally responsive support, families would likely accept services. Community-based participatory methods are effective for eliciting root causes of health inequities in marginalized communities. Outreach to community-based organizations and an inclusive research practice identified social and cultural reasons for low service uptake and provided a pathway for the community to improve services for persons with developmental disabilities.

  11. A Cross-Cultural Comparison of Childhood Developmental Disorders between Schoolchildren in the Netherlands and Greece

    ERIC Educational Resources Information Center

    Manti, Eirini; Scholte, Evert M.; Van Berckelaer-Onnes, Ina A.

    2009-01-01

    The scope of the present study was first to evaluate the cross-cultural reliability and validity of the Social Emotional Questionnaire (SEQ) and second to estimate and compare the prevalence rates of childhood developmental and psychiatric disorders in the general population of young children in the Netherlands and Greece. To this end, the…

  12. Developmental and Cross-Cultural Differences in the Cooperative and Competitive Behavior of Young Children.

    ERIC Educational Resources Information Center

    Madsen, Millard C.

    An experimental task with accompanying apparatus was developed for use in the study of developmental and cultural differences in the cooperative-competitive behavior of children in a small Mexican town and in California. Two groups of 20 Mexican children (aged 7-8 and 10-11), from an elementary school in a town in Baja, California, Mexico, were…

  13. Culture and the Trajectories of Developmental Pathology: Insights from Control and Information Theories.

    PubMed

    Wallace, Rodrick

    2018-06-01

    Cognition in living entities-and their social groupings or institutional artifacts-is necessarily as complicated as their embedding environments, which, for humans, includes a particularly rich cultural milieu. The asymptotic limit theorems of information and control theories permit construction of a new class of empirical 'regression-like' statistical models for cognitive developmental processes, their dynamics, and modes of dysfunction. Such models may, as have their simpler analogs, prove useful in the study and re-mediation of cognitive failure at and across the scales and levels of organization that constitute and drive the phenomena of life. These new models particularly focus on the roles of sociocultural environment and stress, in a large sense, as both trigger for the failure of the regulation of bio-cognition and as 'riverbanks' determining the channels of pathology, with implications across life-course developmental trajectories. We examine the effects of an embedding cultural milieu and its socioeconomic implementations using the 'lenses' of metabolic optimization, control system theory, and an extension of symmetry-breaking appropriate to information systems. A central implication is that most, if not all, human developmental disorders are fundamentally culture-bound syndromes. This has deep implications for both individual treatment and public health policy.

  14. Brain stem slice conditioned medium contains endogenous BDNF and GDNF that affect neural crest boundary cap cells in co-culture.

    PubMed

    Kaiser, Andreas; Kale, Ajay; Novozhilova, Ekaterina; Siratirakun, Piyaporn; Aquino, Jorge B; Thonabulsombat, Charoensri; Ernfors, Patrik; Olivius, Petri

    2014-05-30

    Conditioned medium (CM), made by collecting medium after a few days in cell culture and then re-using it to further stimulate other cells, is a known experimental concept since the 1950s. Our group has explored this technique to stimulate the performance of cells in culture in general, and to evaluate stem- and progenitor cell aptitude for auditory nerve repair enhancement in particular. As compared to other mediums, all primary endpoints in our published experimental settings have weighed in favor of conditioned culture medium, where we have shown that conditioned culture medium has a stimulatory effect on cell survival. In order to explore the reasons for this improved survival we set out to analyze the conditioned culture medium. We utilized ELISA kits to investigate whether brain stem (BS) slice CM contains any significant amounts of brain-derived neurotrophic factor (BDNF) and glial cell derived neurotrophic factor (GDNF). We further looked for a donor cell with progenitor characteristics that would be receptive to BDNF and GDNF. We chose the well-documented boundary cap (BC) progenitor cells to be tested in our in vitro co-culture setting together with cochlear nucleus (CN) of the BS. The results show that BS CM contains BDNF and GDNF and that survival of BC cells, as well as BC cell differentiation into neurons, were enhanced when BS CM were used. Altogether, we conclude that BC cells transplanted into a BDNF and GDNF rich environment could be suitable for treatment of a traumatized or degenerated auditory nerve. Copyright © 2014 Elsevier B.V. All rights reserved.

  15. Developmental milestones among Aboriginal children in Canada

    PubMed Central

    Findlay, Leanne; Kohen, Dafna; Miller, Anton

    2014-01-01

    BACKGROUND: Windows of achievement provide age ranges for the attainment of early developmental skills. Group-specific research is warranted given that development may be influenced by social or cultural factors. OBJECTIVES: To examine developmental milestones for Inuit, Métis and off-reserve First Nation children in Canada, based on developmental domains collected from the 2006 Aboriginal Children’s Survey. Sociodemographic and health predictors of risk for developmental delay were also examined. RESULTS: The ranges in which children achieve certain developmental milestones are presented. Gross motor and self-help skills were found to be achieved earlier (across the three Aboriginal groups), whereas language skills were achieved slightly later than in Canadian children in general. Furthermore, health factors (eg, low birth weight, chronic health conditions) were associated with late achievement of developmental outcomes even when sociodemographic characteristics were considered. CONCLUSIONS: Findings suggest that the timing of milestone achievement may differ for Aboriginal children, highlighting the importance of establishing culturally specific norms and standards rather than relying on those derived from general populations. This information may be useful for practitioners and parents interested in identifying the age ranges for development, as well as age ranges indicating potential for developmental risk and opportunities for early intervention among Aboriginal children. PMID:24855426

  16. Preliminary Study of Realistic Blast Impact on Cultured Brain Slices

    DTIC Science & Technology

    2015-04-01

    and/or multiple impacts in water. 3. Experimental Setup 3.1 The Aquarium Setup A 30.5-cm by 34.5- × 65-cm water-filled polymethylmethacrylate ...sodium bicarbonate PAGE polyacrylamide gel electrophoresis PMMA polymethylmethacrylate RDECOM U.S. Army Research Development and Engineering Command

  17. In vitro studies on the putative function of N-acetylaspartate as an osmoregulator.

    PubMed

    Tranberg, Mattias; Abbas, Abdul-Karim; Sandberg, Mats

    2007-07-01

    Efflux and tissue content of N-acetylaspartate (NAA) and amino acids were evaluated from cultured and acutely prepared hippocampal slices in response to changes in osmolarity. The osmoregulator taurine, but not NAA, was lost from both types of slices after moderate reductions in extracellular osmolarity (-60 mOsm) for 10-48 h. Hypoosmotic shock (-166 mOsm) for 5 min resulted in unselective efflux of several amino acids from acutely prepared slices. Notably, the efflux of taurine, but not NAA, was prominent also after the shock. Efflux of NAA was markedly enhanced by NMDA and high K(+), in particular after the stimulation period. The high K(+)-mediated efflux was decreased by high extracellular osmolarity and a NMDA-receptor antagonist. The results indicate that NAA efflux can be induced by a sudden non-physiological decrease in extracellular osmolarity but not by prolonged more moderate changes in osmolarity. The mechanisms behind the efflux of NAA by high K(+) are complex and may involve both swelling and activation of NMDA-receptors.

  18. Deconstructing brain-derived neurotrophic factor actions in adult brain circuits to bridge an existing informational gap in neuro-cell biology.

    PubMed

    Bowling, Heather; Bhattacharya, Aditi; Klann, Eric; Chao, Moses V

    2016-03-01

    Brain-derived neurotrophic factor (BDNF) plays an important role in neurodevelopment, synaptic plasticity, learning and memory, and in preventing neurodegeneration. Despite decades of investigations into downstream signaling cascades and changes in cellular processes, the mechanisms of how BDNF reshapes circuits in vivo remain unclear. This informational gap partly arises from the fact that the bulk of studies into the molecular actions of BDNF have been performed in dissociated neuronal cultures, while the majority of studies on synaptic plasticity, learning and memory were performed in acute brain slices or in vivo. A recent study by Bowling-Bhattacharya et al., measured the proteomic changes in acute adult hippocampal slices following treatment and reported changes in proteins of neuronal and non-neuronal origin that may in concert modulate synaptic release and secretion in the slice. In this paper, we place these findings into the context of existing literature and discuss how they impact our understanding of how BDNF can reshape the brain.

  19. Neuronal network imaging in acute slices using Ca2+ sensitive bioluminescent reporter.

    PubMed

    Tricoire, Ludovic; Lambolez, Bertrand

    2014-01-01

    Genetically encoded indicators are valuable tools to study intracellular signaling cascades in real time using fluorescent or bioluminescent imaging techniques. Imaging of Ca(2+) indicators is widely used to record transient intracellular Ca(2+) increases associated with bioelectrical activity. The natural bioluminescent Ca(2+) sensor aequorin has been historically the first Ca(2+) indicator used to address biological questions. Aequorin imaging offers several advantages over fluorescent reporters: it is virtually devoid of background signal; it does not require light excitation and interferes little with intracellular processes. Genetically encoded sensors such as aequorin are commonly used in dissociated cultured cells; however it becomes more challenging to express them in differentiated intact specimen such as brain tissue. Here we describe a method to express a GFP-aequorin (GA) fusion protein in pyramidal cells of neocortical acute slices using recombinant Sindbis virus. This technique allows expressing GA in several hundreds of neurons on the same slice and to perform the bioluminescence recording of Ca(2+) transients in single neurons or multiple neurons simultaneously.

  20. Bidirectional Signaling of Neuregulin-2 Mediates Formation of GABAergic Synapses and Maturation of Glutamatergic Synapses in Newborn Granule Cells of Postnatal Hippocampus.

    PubMed

    Lee, Kyu-Hee; Lee, Hyunsu; Yang, Che Ho; Ko, Jeong-Soon; Park, Chang-Hwan; Woo, Ran-Sook; Kim, Joo Yeon; Sun, Woong; Kim, Joung-Hun; Ho, Won-Kyung; Lee, Suk-Ho

    2015-12-16

    Expression of neuregulin-2 (NRG2) is intense in a few regions of the adult brain where neurogenesis persists; however, little is understood about its role in developments of newborn neurons. To study the role of NRG2 in synaptogenesis at different developmental stages, newborn granule cells in rat hippocampal slice cultures were labeled with retrovirus encoding tetracycline-inducible microRNA targeting NRG2 and treated with doxycycline (Dox) at the fourth or seventh postinfection day (dpi). The developmental increase of GABAergic postsynaptic currents (GPSCs) was suppressed by the early Dox treatment (4 dpi), but not by late treatment (7 dpi). The late Dox treatment was used to study the effect of NRG2 depletion specific to excitatory synaptogenesis. The Dox effect on EPSCs emerged 4 d after the impairment in dendritic outgrowth became evident (10 dpi). Notably, Dox treatment abolished the developmental increases of AMPA-receptor mediated EPSCs and the AMPA/NMDA ratio, indicating impaired maturation of glutamatergic synapses. In contrast to GPSCs, Dox effects on EPSCs and dendritic growth were independent of ErbB4 and rescued by concurrent overexpression of NRG2 intracellular domain. These results suggest that forward signaling of NRG2 mediates GABAergic synaptogenesis and its reverse signaling contributes to dendritic outgrowth and maturation of glutamatergic synapses. The hippocampal dentate gyrus is one of special brain regions where neurogenesis persists throughout adulthood. Synaptogenesis is a critical step for newborn neurons to be integrated into preexisting neural network. Because neuregulin-2 (NRG2), a growth factor, is intensely expressed in these regions, we investigated whether it plays a role in synaptogenesis and dendritic growth. We found that NRG2 has dual roles in the development of newborn neurons. For GABAergic synaptogenesis, the extracellular domain of NRG2 acts as a ligand for a receptor on GABAergic neurons. In contrast, its intracellular domain was essential for dendritic outgrowth and glutamatergic synapse maturation. These results imply that NRG2 may play a critical role in network integration of newborn neurons. Copyright © 2015 the authors 0270-6474/15/3516480-15$15.00/0.

  1. Developmental Transition of Motherhood: Treating Postpartum Depression Using a Feminist Approach

    ERIC Educational Resources Information Center

    Davis-Gage, Darcie; Kettmann, Julie Jenks; Moel, Joy

    2010-01-01

    During the developmental lifeline for women, some individuals are affected by postpartum depression. This article describes the treatment of a Latina woman experiencing postpartum depression. The authors illustrate the feminist approach using counseling interventions that incorporate the client's developmental level, cultural background, and…

  2. Discerning cultural identification from a thinly sliced behavioral sample.

    PubMed

    Hamamura, Takeshi; Li, Liman Man Wai

    2012-12-01

    This research examined whether individual differences in cultural identification can be discerned at zero acquaintance. This issue was examined in Hong Kong, where the idiosyncrasy of cultural identification is a salient social-psychological issue. The participants were able to perceive accurately the targets' identification with Western culture from a video clip and from a still image. Findings also indicated that a stereotype of Western cultural identity (i.e., extraversion and particular hairstyle) facilitated these perceptions. Specifically, (a) the participants with a stronger stereotype were more accurate in perceiving Western cultural identification, (b) the targets who were experimentally manipulated to appear extraverted were rated as more strongly identifying with Western culture, and (c) the participants relatively unfamiliar with these stereotypes did not correctly perceive Western cultural identification. Implications of these findings on research on multiculturalism are also discussed.

  3. Why developmental psychology is incomplete without comparative and cross-cultural perspectives.

    PubMed

    Nielsen, Mark; Haun, Daniel

    2016-01-19

    As a discipline, developmental psychology has a long history of relying on animal models and data collected among distinct cultural groups to enrich and inform theories of the ways social and cognitive processes unfold through the lifespan. However, approaches that draw together developmental, cross-cultural and comparative perspectives remain rare. The need for such an approach is reflected in the papers by Heyes (2015 Phil. Trans. R. Soc. B 371, 20150069. (doi:10.1098/rstb.2015.0069)), Schmelz & Call (2015 Phil. Trans. R. Soc. B 371, 20150067. (doi:10.1098/rstb.2015.0067)) and Keller (2015 Phil. Trans. R. Soc. B 371, 20150070. (doi:10.1098/rstb.2015.0070)) in this theme issue. Here, we incorporate these papers into a review of recent research endeavours covering a range of core aspects of social cognition, including social learning, cooperation and collaboration, prosociality, and theory of mind. In so doing, we aim to highlight how input from comparative and cross-cultural empiricism has altered our perspectives of human development and, in particular, led to a deeper understanding of the evolution of the human cultural mind. © 2015 The Author(s).

  4. Why developmental psychology is incomplete without comparative and cross-cultural perspectives

    PubMed Central

    Nielsen, Mark; Haun, Daniel

    2016-01-01

    As a discipline, developmental psychology has a long history of relying on animal models and data collected among distinct cultural groups to enrich and inform theories of the ways social and cognitive processes unfold through the lifespan. However, approaches that draw together developmental, cross-cultural and comparative perspectives remain rare. The need for such an approach is reflected in the papers by Heyes (2015 Phil. Trans. R. Soc. B 371, 20150069. (doi:10.1098/rstb.2015.0069)), Schmelz & Call (2015 Phil. Trans. R. Soc. B 371, 20150067. (doi:10.1098/rstb.2015.0067)) and Keller (2015 Phil. Trans. R. Soc. B 371, 20150070. (doi:10.1098/rstb.2015.0070)) in this theme issue. Here, we incorporate these papers into a review of recent research endeavours covering a range of core aspects of social cognition, including social learning, cooperation and collaboration, prosociality, and theory of mind. In so doing, we aim to highlight how input from comparative and cross-cultural empiricism has altered our perspectives of human development and, in particular, led to a deeper understanding of the evolution of the human cultural mind. PMID:26644590

  5. The influence of developmental, education, and mentorship experiences on career paths in cultural psychiatry.

    PubMed

    Boehnlein, James K

    2011-04-01

    Career paths in cultural psychiatry and other areas of medicine often are influenced by a combination of developmental, academic and training experiences. One of the satisfactions for the cultural psychiatrist is the opportunity to integrate lifelong interests that are not central to medical training, such as anthropology, philosophy, history or geography into one's daily work and career path, and this is not necessarily just applicable to clinical work or research. A background in social sciences can be very helpful in navigating the political and interpersonal challenges of working in an academic medical center or designing effective education programs across the career spectrum in medicine. This article traces the development of my career in academic cultural psychiatry and illustrates the way my clinical and academic interests were influenced both by early developmental and educational experiences and by positive career experiences as a clinician, teacher and researcher in the context of effective mentorship and professional peer relationships. The future of cultural psychiatry is exciting and its continued growth will be dependent on effective nurturance of young physicians who have a broad vision of the important place of cultural psychiatry and how it influences medicine, mental health, and society at large.

  6. Assessing archetypes of organizational culture based on the Competing Values Framework: the experimental use of the framework in Japanese neonatal intensive care units

    PubMed Central

    Sasaki, Hatoko; Yonemoto, Naohiro; Mori, Rintaro; Nishida, Toshihiko; Kusuda, Satoshi; Nakayama, Takeo

    2017-01-01

    Abstract Objective To assess organizational culture in neonatal intensive care units (NICUs) in Japan. Design Cross-sectional survey of organizational culture. Setting Forty NICUs across Japan. Participants Physicians and nurses who worked in NICUs (n = 2006). Main Outcome Measures The Competing Values Framework (CVF) was used to assess the organizational culture of the study population. The 20-item CVF was divided into four culture archetypes: Group, Developmental, Hierarchical and Rational. We calculated geometric means (gmean) and 95% bootstrap confidence intervals of the individual dimensions by unit and occupation. The median number of staff, beds, physicians’ work hours and work engagement were also calculated to examine the differences by culture archetypes. Results Group (gmean = 34.6) and Hierarchical (gmean = 31.7) culture archetypes were higher than Developmental (gmean = 16.3) and Rational (gmean = 17.4) among physicians as a whole. Hierarchical (gmean = 36.3) was the highest followed by Group (gmean = 25.8), Developmental (gmean = 16.3) and Rational (gmean = 21.7) among nurses as a whole. Units with dominant Hierarchical culture had a slightly higher number of physicians (median = 7) than dominant Group culture (median = 6). Units with dominant Group culture had a higher number of beds (median = 12) than dominant Hierarchical culture (median = 9) among physicians. Nurses from units with a dominant Group culture (median = 2.8) had slightly higher work engagement compared with those in units with a dominant Hierarchical culture (median = 2.6). Conclusions Our findings revealed that organizational culture in NICUs varies depending on occupation and group size. Group and Hierarchical cultures predominated in Japanese NICUs. Assessing organizational culture will provide insights into the perceptions of unit values to improve quality of care. PMID:28371865

  7. Assessing archetypes of organizational culture based on the Competing Values Framework: the experimental use of the framework in Japanese neonatal intensive care units.

    PubMed

    Sasaki, Hatoko; Yonemoto, Naohiro; Mori, Rintaro; Nishida, Toshihiko; Kusuda, Satoshi; Nakayama, Takeo

    2017-06-01

    To assess organizational culture in neonatal intensive care units (NICUs) in Japan. Cross-sectional survey of organizational culture. Forty NICUs across Japan. Physicians and nurses who worked in NICUs (n = 2006). The Competing Values Framework (CVF) was used to assess the organizational culture of the study population. The 20-item CVF was divided into four culture archetypes: Group, Developmental, Hierarchical and Rational. We calculated geometric means (gmean) and 95% bootstrap confidence intervals of the individual dimensions by unit and occupation. The median number of staff, beds, physicians' work hours and work engagement were also calculated to examine the differences by culture archetypes. Group (gmean = 34.6) and Hierarchical (gmean = 31.7) culture archetypes were higher than Developmental (gmean = 16.3) and Rational (gmean = 17.4) among physicians as a whole. Hierarchical (gmean = 36.3) was the highest followed by Group (gmean = 25.8), Developmental (gmean = 16.3) and Rational (gmean = 21.7) among nurses as a whole. Units with dominant Hierarchical culture had a slightly higher number of physicians (median = 7) than dominant Group culture (median = 6). Units with dominant Group culture had a higher number of beds (median = 12) than dominant Hierarchical culture (median = 9) among physicians. Nurses from units with a dominant Group culture (median = 2.8) had slightly higher work engagement compared with those in units with a dominant Hierarchical culture (median = 2.6). Our findings revealed that organizational culture in NICUs varies depending on occupation and group size. Group and Hierarchical cultures predominated in Japanese NICUs. Assessing organizational culture will provide insights into the perceptions of unit values to improve quality of care. © The Author 2017. Published by Oxford University Press in association with the International Society for Quality in Health Care

  8. The Ultimate Developmental Task in Adolescent Literature.

    ERIC Educational Resources Information Center

    Abrahamson, Richard F.

    The significance of adolescent literature has been judged on the basis of the developmental tasks encountered by the main character. One writer has identified eight developmental tasks that teenagers must undertake as they move toward adulthood: discovering one's sex role in our culture, developing relationships with peers, achieving an easy…

  9. Integrated organotypic slice cultures and RT-QuIC (OSCAR) assay: implications for translational discovery in protein misfolding diseases

    USDA-ARS?s Scientific Manuscript database

    Protein misfolding is a key pathological event in neurodegenerative diseases like prion diseases, synucleinopathies, and tauopathies that are collectively termed protein misfolding disorders (PMD). Prions are a prototypic model to study protein aggregation biology and therapeutic development. Attemp...

  10. REGULATION OF BRAIN-DERIVED NEUROTROPHIC FACTOR MESSENGER RNA LEVELS IN AVIAN HYPOTHALAMIC SLICE CULTURES. (R825294)

    EPA Science Inventory

    Mechanisms regulating the expression of brain-derived neurotrophic factor, a member of the neurotrophin family, have been extensively studied in the rat cerebral cortex, hippocampus and cerebellum. In contrast, little is known regarding the regulation of this growth factor in ...

  11. EFFECT OF PHENYTOIN ON GENE EXPRESSION, OXIDATIVE DAMAGE AND CELL VIABILITY OF CULTURED HUMAN FETAL LIVER SLICES. (R827441)

    EPA Science Inventory

    The perspectives, information and conclusions conveyed in research project abstracts, progress reports, final reports, journal abstracts and journal publications convey the viewpoints of the principal investigator and may not represent the views and policies of ORD and EPA. Concl...

  12. European-French Cross-Cultural Adaptation of the Developmental Coordination Disorder Questionnaire and Pretest in French-Speaking Switzerland.

    PubMed

    Ray-Kaeser, Sylvie; Satink, Ton; Andresen, Mette; Martini, Rose; Thommen, Evelyne; Bertrand, Anne Martine

    2015-05-01

    The Developmental Coordination Disorder Questionnaire (DCDQ'07) is a Canadian-English instrument recommended for screening children aged 5 to 15 years who are at risk for developmental coordination disorder. While a Canadian-French version of the DCDQ'07 presently exists, a European-French version does not. To produce a cross-cultural adaptation of the DCDQ'07 for use in areas of Europe where French is spoken and to test its cultural relevance in French-speaking Switzerland. Cross-cultural adaptation was done using established guidelines. Cultural relevance was analyzed with cognitive interviews of thirteen parents of children aged 5.0 to 14.6 years (mean age: 8.5 years, SD = 3.4), using think-aloud and probing techniques. Cultural and linguistic differences were noted between the European-French, the Canadian-French, and the original versions of the DCDQ'07. Despite correct translation and expert committee review, cognitive interviews revealed that certain items of the European-French version were unclear or misinterpreted and further modifications were needed. After rewording items as a result of the outcomes of the cognitive interview, the European-French version of the DCDQ'07 is culturally appropriate for use in French-speaking Switzerland. Further studies are necessary to determine its psychometric properties.

  13. Study on tissue culture for Gelidium seedling

    NASA Astrophysics Data System (ADS)

    Pei, Lu-Qing; Luo, Qi-Jun; Fei, Zhi-Qing; Ma, Bin

    1996-06-01

    As seedling culture is a crucial factor for successful cultivation of Gelidium, the authors researched tissue culture technology for producing seedlings. The morphogeny and experimental ecology were observed and studied fully in 2 5 mm isolated tissue fragments. Regeneration, appearance of branching creepers and attaching structure and new erect seedlings production and development were studied. Fragments were sown on bamboo slice and vinylon rope. The seedlings were cultured 20 30 days indoor, then cultured in the sea, where the density of erect seedlings was 3 19 seedlings/cm2, growth rate was 3.84% day. The frond arising from seedlings directly was up to 10 cm per year. The ecological conditions for regenerated seedlings are similar to the natural ones. The regenerated seedlings are suitable for raft culture in various sea areas.

  14. Universal, developmental, and variable aspects of young children's play: a cross-cultural comparison of pretending at home.

    PubMed

    Haight, W L; Wang, X L; Fung, H H; Williams, K; Mintz, J

    1999-01-01

    Using longitudinal data from five Irish American families in the United States and nine Chinese families in Taiwan, in conjunction with an emerging body of evidence in the cultural psychology literature, we propose universal, culturally variable, and developmental dimensions of young children's pretend play. Possible universal dimensions include the use of objects, and the predominantly social nature of pretend play. Developmental dimensions include increases in the proportion of social pretend play initiated by the child, the proportion of partner initiations elaborated upon by the child, and caregivers' use of pretend play initiations to serve other, nonplay social functions. Culturally variable dimensions include the centrality of objects, the participation of specific play partners, the extent of child initiations of social pretend play with caregivers, the various functions of social pretend play in interaction, and specific themes. These findings raise the theoretical issue of how universal and variable dimensions of pretend play interact in specific communities to create distinctive development pathways.

  15. AN ASSESSMENT OF CULTURAL VALUES AND RESIDENT-CENTERED CULTURE CHANGE IN US NURSING FACILITIES

    PubMed Central

    Banaszak-Holl, Jane; Castle, Nicholas G.; Lin, Michael; Spreitzer, Gretchen

    2012-01-01

    Background Culture Change initiatives propose to improve care by addressing the lack of managerial supports and prevalent stressful work environments in the industry; however, little is known about how Culture Change facilities differ from facilities in the industry that have not chosen to affiliate with the resident-centered care movements. Purpose To evaluate representation of organizational culture values within a random sample of U.S. nursing home facilities using the Competing Values Framework (CVF) and to determine whether organizational values are related to membership in resident-centered Culture Change initiatives. Design and Methods We collected reports of cultural values using a well-established CVF instrument in a random survey of facility administrators and directors of nursing within all states. We received responses from 57% of the facilities that were mailed the survey. Directors of nursing and administrators did not vary significantly in their reports of culture and facility measures combine their responses. Findings Nursing facilities favored market-focused cultural values on average and developmental values, key to innovation, were the least common across all nursing homes. Approximately 17% of facilities reported all cultural values were strong within their facilities. Only high developmental cultural values were linked to participation in culture change initiatives. Culture Change facilities were not different from non-Culture Change facilities in the promotion of employee focus as organizational culture, as is emphasized in group culture values. Likewise, Culture Change facilities were also not any more likely to have hierarchical or market foci than non-Culture Change facilities. Practice Implications Our results counter the argument that Culture Change facilities have a stronger internal employee focus than facilities more generally but does show that Culture Change facilities report stronger developmental cultures than non-Culture Change facilities, which indicates a potential to be innovative in their strategies. Facilities are culturally ready to become resident-centered and may face other barriers to adopting these practices. PMID:22936002

  16. An assessment of cultural values and resident-centered culture change in U.S. nursing facilities.

    PubMed

    Banaszak-Holl, Jane; Castle, Nicholas G; Lin, Michael; Spreitzer, Gretchen

    2013-01-01

    Culture change initiatives propose to improve care by addressing the lack of managerial supports and prevalent stressful work environments in the industry; however, little is known about how culture change facilities differ from facilities in the industry that have not chosen to affiliate with the resident-centered care movements. The aim of this study was to evaluate representation of organizational culture values within a random sample of U.S. nursing home facilities using the competing values framework and to determine whether organizational values are related to membership in resident-centered culture change initiatives. We collected reports of cultural values using a well-established competing values framework instrument in a random survey of facility administrators and directors of nursing within all states. We received responses from 57% of the facilities that were mailed the survey. Directors of nursing and administrators did not differ significantly in their reports of culture and facility measures combined their responses. Nursing facilities favored market-focused cultural values on average, and developmental values, key to innovation, were the least common across all nursing homes. Approximately 17% of the facilities reported that all cultural values were strong within their facilities. Only high developmental cultural values were linked to participation in culture change initiatives. Culture change facilities were not different from non-culture change facilities in the promotion of employee focus as organizational culture, as emphasized in group culture values. Likewise, culture change facilities were also not more likely to have hierarchical or market foci than non-culture change facilities. Our results counter the argument that culture change facilities have a stronger internal employee focus than facilities more generally but do show that culture change facilities report stronger developmental cultures than non-culture change facilities, which indicates a potential to be innovative in their strategies. Facilities are culturally ready to become resident centered and may face other barriers to adopting these practices.

  17. Priming 3D cultures of human mesenchymal stromal cells toward cartilage formation via developmental pathways.

    PubMed

    Centola, Matteo; Tonnarelli, Beatrice; Schären, Stefan; Glaser, Nicolas; Barbero, Andrea; Martin, Ivan

    2013-11-01

    The field of regenerative medicine has increasingly recognized the importance to be inspired by developmental processes to identify signaling pathways crucial for 3D organogenesis and tissue regeneration. Here, we aimed at recapitulating the first events occurring during limb development (ie, cell condensation and expansion of an undifferentiated mesenchymal cell population) to prime 3D cultures of human bone marrow-derived mesenchymal stromal/stem cells (hBM-MSC) toward the chondrogenic route. Based on embryonic development studies, we hypothesized that Wnt3a and fibroblast growth factor 2 (FGF2) induce hBM-MSC to proliferate in 3D culture as an undifferentiated pool of progenitors (defined by clonogenic capacity and expression of typical markers), retaining chondrogenic potential upon induction by suitable morphogens. hBM-MSC were responsive to Wnt signaling in 3D pellet culture, as assessed by significant upregulation of main target genes and increase of unphosphorylated β-catenin levels. Wnt3a was able to induce a five-fold increase in the number of proliferating hBM-MSC (6.4% vs. 1.3% in the vehicle condition), although total DNA content of the 3D construct was decreasing over time. Preconditioning with Wnt3a improved transforming growth factor-β1 mediated chondrogenesis (30% more glycosaminoglycans/cell in average). In contrast to developmental and 2D MSC culture models, FGF2 antagonized the Wnt-mediated effects. Interestingly, the CD146⁺ subpopulation was found to be more responsive to Wnt3a. The presented data indicate a possible strategy to prime 3D cultures of hBM-MSC by invoking a "developmental engineering" approach. The study also identifies some opportunities and challenges to cross-fertilize skeletal development models and 3D hBM-MSC culture systems.

  18. Systems analysis of a maize leaf developmental gradient redefines the current C4 model and provides candidates for regulation.

    PubMed

    Pick, Thea R; Bräutigam, Andrea; Schlüter, Urte; Denton, Alisandra K; Colmsee, Christian; Scholz, Uwe; Fahnenstich, Holger; Pieruschka, Roland; Rascher, Uwe; Sonnewald, Uwe; Weber, Andreas P M

    2011-12-01

    We systematically analyzed a developmental gradient of the third maize (Zea mays) leaf from the point of emergence into the light to the tip in 10 continuous leaf slices to study organ development and physiological and biochemical functions. Transcriptome analysis, oxygen sensitivity of photosynthesis, and photosynthetic rate measurements showed that the maize leaf undergoes a sink-to-source transition without an intermediate phase of C(3) photosynthesis or operation of a photorespiratory carbon pump. Metabolome and transcriptome analysis, chlorophyll and protein measurements, as well as dry weight determination, showed continuous gradients for all analyzed items. The absence of binary on-off switches and regulons pointed to a morphogradient along the leaf as the determining factor of developmental stage. Analysis of transcription factors for differential expression along the leaf gradient defined a list of putative regulators orchestrating the sink-to-source transition and establishment of C(4) photosynthesis. Finally, transcriptome and metabolome analysis, as well as enzyme activity measurements, and absolute quantification of selected metabolites revised the current model of maize C(4) photosynthesis. All data sets are included within the publication to serve as a resource for maize leaf systems biology.

  19. Systems Analysis of a Maize Leaf Developmental Gradient Redefines the Current C4 Model and Provides Candidates for Regulation[W][OA

    PubMed Central

    Pick, Thea R.; Bräutigam, Andrea; Schlüter, Urte; Denton, Alisandra K.; Colmsee, Christian; Scholz, Uwe; Fahnenstich, Holger; Pieruschka, Roland; Rascher, Uwe; Sonnewald, Uwe; Weber, Andreas P.M.

    2011-01-01

    We systematically analyzed a developmental gradient of the third maize (Zea mays) leaf from the point of emergence into the light to the tip in 10 continuous leaf slices to study organ development and physiological and biochemical functions. Transcriptome analysis, oxygen sensitivity of photosynthesis, and photosynthetic rate measurements showed that the maize leaf undergoes a sink-to-source transition without an intermediate phase of C3 photosynthesis or operation of a photorespiratory carbon pump. Metabolome and transcriptome analysis, chlorophyll and protein measurements, as well as dry weight determination, showed continuous gradients for all analyzed items. The absence of binary on–off switches and regulons pointed to a morphogradient along the leaf as the determining factor of developmental stage. Analysis of transcription factors for differential expression along the leaf gradient defined a list of putative regulators orchestrating the sink-to-source transition and establishment of C4 photosynthesis. Finally, transcriptome and metabolome analysis, as well as enzyme activity measurements, and absolute quantification of selected metabolites revised the current model of maize C4 photosynthesis. All data sets are included within the publication to serve as a resource for maize leaf systems biology. PMID:22186372

  20. Stimulation of brain glucose uptake by cannabinoid CB2 receptors and its therapeutic potential in Alzheimer's disease.

    PubMed

    Köfalvi, Attila; Lemos, Cristina; Martín-Moreno, Ana M; Pinheiro, Bárbara S; García-García, Luis; Pozo, Miguel A; Valério-Fernandes, Ângela; Beleza, Rui O; Agostinho, Paula; Rodrigues, Ricardo J; Pasquaré, Susana J; Cunha, Rodrigo A; de Ceballos, María L

    2016-11-01

    Cannabinoid CB2 receptors (CB2Rs) are emerging as important therapeutic targets in brain disorders that typically involve neurometabolic alterations. We here addressed the possible role of CB2Rs in the regulation of glucose uptake in the mouse brain. To that aim, we have undertaken 1) measurement of (3)H-deoxyglucose uptake in cultured cortical astrocytes and neurons and in acute hippocampal slices; 2) real-time visualization of fluorescently labeled deoxyglucose uptake in superfused hippocampal slices; and 3) in vivo PET imaging of cerebral (18)F-fluorodeoxyglucose uptake. We now show that both selective (JWH133 and GP1a) as well as non-selective (WIN55212-2) CB2R agonists, but not the CB1R-selective agonist, ACEA, stimulate glucose uptake, in a manner that is sensitive to the CB2R-selective antagonist, AM630. Glucose uptake is stimulated in astrocytes and neurons in culture, in acute hippocampal slices, in different brain areas of young adult male C57Bl/6j and CD-1 mice, as well as in middle-aged C57Bl/6j mice. Among the endocannabinoid metabolizing enzymes, the selective inhibition of COX-2, rather than that of FAAH, MAGL or α,βDH6/12, also stimulates the uptake of glucose in hippocampal slices of middle-aged mice, an effect that was again prevented by AM630. However, we found the levels of the endocannabinoid, anandamide reduced in the hippocampus of TgAPP-2576 mice (a model of β-amyloidosis), and likely as a consequence, COX-2 inhibition failed to stimulate glucose uptake in these mice. Together, these results reveal a novel general glucoregulatory role for CB2Rs in the brain, raising therapeutic interest in CB2R agonists as nootropic agents. Copyright © 2016 The Authors. Published by Elsevier Ltd.. All rights reserved.

  1. The Experiences and Perspectives of Japanese Substitute Caregivers and Maltreated Children: A Cultural-Developmental Approach to Child Welfare Practice

    ERIC Educational Resources Information Center

    Bamba, Sachiko

    2010-01-01

    This article describes the experiences and perspectives of child welfare workers and maltreated children living in Japanese state care. Japanese adults emphasize supporting children's emotional well-being and empowerment through developmentally and ecologically focused socialization strategies. One developmental goal articulated by caregivers of…

  2. The Arts as a Venue for Developmental Science: Realizing a Latent Opportunity

    ERIC Educational Resources Information Center

    Goldstein, Thalia R.; Lerner, Matthew D.; Winner, Ellen

    2017-01-01

    Children in all cultures readily engage in artistic activities, yet the arts (dance, drama, drawing, and music) have traditionally been marginal topics in the discipline of developmental science. We argue that developmental psychologists cannot afford to ignore such naturalistic activities that involve so many basic phenomena--attention,…

  3. Parent socialization effects in different cultures: significance of directive parenting.

    PubMed

    Sorkhabi, Nadia

    2012-06-01

    In this article, the controversy of divergent findings in research on parental socialization effects in different cultures is addressed. Three explanations intended to address divergent findings of socialization effects in different cultures, as advanced by researchers who emphasize cultural differences, are discussed. These include cultural differences in socialization values and goals of parents, parental emotional and cognitive characteristics associated with parenting styles, and adolescents' interpretations or evaluations of their parents' parenting styles. The empirical evidence for and against each of these arguments is examined and an alternative paradigm for understanding and empirical study of developmental outcomes associated with parenting styles in different cultures is suggested. Baumrind's directive parenting style is presented as an alternative to the authoritarian parenting style in understanding the positive developmental effects associated with "strict" parenting in cultures said to have a collectivist orientation. Directions for research on the three explanations are mentioned.

  4. Process-product dynamics: the role of Otherness in cultural cultivation.

    PubMed

    Lyra, Maria C D P

    2014-06-01

    Carriere (2013) presents a stimulating perspective on the cultural phenomena aiming to recover the role of the external products of culture to imbalance the currently popular emphasis on subject's process of cultivation highlighted by semiotic developmental cultural psychology. The excessive focus on subject's internal processes dismissing a better consideration of products of culture and the compelling objective realities of other dimensions of culture are pointed out. By this way the author's proposes a better dialogue with others perspectives on (cross)cultural psychology. These arguments are analyzed through a closer consideration of I-Other perennial movement. A dialogical view of process-product dynamics is then proposed. The role of Otherness--the one that (partially)shares and the one as witness, approving or disapproving subject's products of cultivation--is discussed through the analysis of a concrete episode of the cultivation of the subject. It is concluded that a semiotic developmental cultural psychology and (cross) cultural psychology have different objects of knowledge comprising distinct interests and research fields.

  5. Discovery of Hyperpolarized Molecular Imaging Biomarkers in a Novel Prostate Tissue Slice Culture Model

    DTIC Science & Technology

    2011-06-01

    we tested was the addition ofN- acetylcysteine (NAC) to the medium. NAC protects cells from oxidative damage, and it is possible that our inability...of tissue procurement and spectral studies • Obtained preliminary data that an anti-oxidant, N- acetylcysteine , might extend longevity of TSCs

  6. Cultural Differences in Sleeping Practices--Helping Early Childhood Educators Understand.

    ERIC Educational Resources Information Center

    Gonzalez-Mena, Janet; Bhavnagri, Navaz Peshotan

    2001-01-01

    Discusses cultural differences in sleeping practices, focusing on how child caregivers can provide developmentally appropriate and culturally sensitive care. Describes co-sleeping as an accepted practice in many cultures with several benefits. Discusses the role of cultural values, beliefs, priorities, and goals and the importance of…

  7. Fast two-photon imaging of subcellular voltage dynamics in neuronal tissue with genetically encoded indicators

    PubMed Central

    Chamberland, Simon; Yang, Helen H; Pan, Michael M; Evans, Stephen W; Guan, Sihui; Chavarha, Mariya; Yang, Ying; Salesse, Charleen; Wu, Haodi; Wu, Joseph C; Clandinin, Thomas R; Toth, Katalin; Lin, Michael Z; St-Pierre, François

    2017-01-01

    Monitoring voltage dynamics in defined neurons deep in the brain is critical for unraveling the function of neuronal circuits but is challenging due to the limited performance of existing tools. In particular, while genetically encoded voltage indicators have shown promise for optical detection of voltage transients, many indicators exhibit low sensitivity when imaged under two-photon illumination. Previous studies thus fell short of visualizing voltage dynamics in individual neurons in single trials. Here, we report ASAP2s, a novel voltage indicator with improved sensitivity. By imaging ASAP2s using random-access multi-photon microscopy, we demonstrate robust single-trial detection of action potentials in organotypic slice cultures. We also show that ASAP2s enables two-photon imaging of graded potentials in organotypic slice cultures and in Drosophila. These results demonstrate that the combination of ASAP2s and fast two-photon imaging methods enables detection of neural electrical activity with subcellular spatial resolution and millisecond-timescale precision. DOI: http://dx.doi.org/10.7554/eLife.25690.001 PMID:28749338

  8. Electroosmotic perfusion of tissue: sampling the extracellular space and quantitative assessment of membrane-bound enzyme activity in organotypic hippocampal slice cultures

    PubMed Central

    Ou, Yangguang; Wu, Juanfang; Sandberg, Mats

    2014-01-01

    This review covers recent advances in sampling fluid from the extracellular space of brain tissue by electroosmosis (EO). Two techniques, EO sampling with a single fused-silica capillary and EO push–pull perfusion, have been developed. These tools were used to investigate the function of membrane-bound enzymes with outward-facing active sites, or ectoenzymes, in modulating the activity of the neuropeptides leu-enkephalin and galanin in organotypic-hippocampal-slice cultures (OHSCs). In addition, the approach was used to determine the endogenous concentration of a thiol, cysteamine, in OHSCs. We have also investigated the degradation of coenzyme A in the extracellular space. The approach provides information on ectoenzyme activity, including Michaelis constants, in tissue, which, as far as we are aware, has not been done before. On the basis of computational evidence, EO push–pull perfusion can distinguish ectoenzyme activity with a ~100 µm spatial resolution, which is important for studies of enzyme kinetics in adjacent regions of the rat hippocampus. PMID:25168111

  9. Ganando Confianza: Research Focus Groups with Immigrant Mexican Mothers

    PubMed Central

    Hausmann-Stabile, Carolina; Zayas, Luis H.; Runes, Sandra; Abenis-Cintron, Anna; Calzada, Esther

    2014-01-01

    Immigrant families with children with developmental disabilities must be served using culturally sensitive approaches to service and research to maximize treatment benefits. In an effort to better understand cultural issues relevant to the provision of parenting programs for immigrant Mexican mothers of children with developmental disabilities, we conducted sustained focus groups through which we could learn more about our participants and thereby improve services. This paper reports on the challenges and lessons learned from these groups. We characterize the key lessons as (a) recruitment and retention is more than agreement to participate; (b) confidentiality is not just a word but an activity; (c) the complicated nature of language; (d) cultural norms shape the group process; (e) appreciating the value of taking time; and (f) gender issues and group interaction. Service providers and researchers who work with Mexican families may benefit from our experiences as they promote and develop programs and projects in the developmental disabilities field. PMID:25674353

  10. Ganando Confianza: Research Focus Groups with Immigrant Mexican Mothers.

    PubMed

    Hausmann-Stabile, Carolina; Zayas, Luis H; Runes, Sandra; Abenis-Cintron, Anna; Calzada, Esther

    2011-03-01

    Immigrant families with children with developmental disabilities must be served using culturally sensitive approaches to service and research to maximize treatment benefits. In an effort to better understand cultural issues relevant to the provision of parenting programs for immigrant Mexican mothers of children with developmental disabilities, we conducted sustained focus groups through which we could learn more about our participants and thereby improve services. This paper reports on the challenges and lessons learned from these groups. We characterize the key lessons as (a) recruitment and retention is more than agreement to participate; (b) confidentiality is not just a word but an activity; (c) the complicated nature of language; (d) cultural norms shape the group process; (e) appreciating the value of taking time; and (f) gender issues and group interaction. Service providers and researchers who work with Mexican families may benefit from our experiences as they promote and develop programs and projects in the developmental disabilities field.

  11. The Efficacy of Arabic Version of the Developmental Assessment of Young Children Second Edition (DAYC-2) Scale in Detecting Developmental Delay among Jordanian Children Aged Birth to 71 Months

    ERIC Educational Resources Information Center

    Saleh, Rawan M. Abu; Smadi, Jamil M.

    2017-01-01

    This study aimed to assess the efficacy of the developmental assessment of young children second edition (DAYC-2) Scale in detecting Developmental Delay among Jordanian children aged birth to 71 months. Firstly, the scale was translated and reviewed for language and cultural appropriateness. Secondly, the Arabic Jordanian version of the scale was…

  12. Endogenous Nod-Factor-Like Signal Molecules Promote Early Somatic Embryo Development in Norway Spruce1

    PubMed Central

    Dyachok, Julia V.; Wiweger, Malgorzata; Kenne, Lennart; von Arnold, Sara

    2002-01-01

    Embryogenic cultures of Norway spruce (Picea abies) are composed of pro-embryogenic masses (PEMs) and somatic embryos of various developmental stages. Auxin is important for PEM formation and proliferation. In this report we show that depletion of auxin blocks PEM development and causes large-scale cell death. Extracts of the media conditioned by embryogenic cultures stimulate development of PEM aggregates in auxin-deficient cultures. Partial characterization of the conditioning factor shows that it is a lipophilic, low-molecular-weight molecule, which is sensitive to chitinase and contains GlcNAc residues. On the basis of this information, we propose that the factor is a lipophilic chitin oligosaccharide (LCO). The amount of LCO correlates to the developmental stages of PEMs and embryos, with the highest level in the media conditioned by developmentally blocked cultures. LCO is not present in nonembryogenic cultures. Cell death, induced by withdrawal of auxin, is suppressed by extra supply of endogenous LCO or Nod factor from Rhizobium sp. NGR234. The effect can be mimicked by a chitotetraose or chitinase from Streptomyces griseus. Taken together, our data suggest that endogenous LCO acts as a signal molecule stimulating PEM and early embryo development in Norway spruce. PMID:11842156

  13. Novel engineered tendon-fibrocartilage-bone composite with cyclic tension for rotator cuff repair.

    PubMed

    Liu, Qian; Hatta, Taku; Qi, Jun; Liu, Haoyu; Thoreson, Andrew R; Amadio, Peter C; Moran, Steven L; Steinmann, Scott P; Gingery, Anne; Zhao, Chunfeng

    2018-05-15

    Surgical repair of rotator cuff tears presents a significant clinical challenge with high failure rates and inferior functional outcomes. Graft augmentation improves repair outcomes, however currently available grafting materials have limitations. While cell-seeded decellularized tendon slices may facilitate cell infiltration, promote tendon incorporation and preserve original mechanical strength, the unique fibrocartilage zone is yet to be successfully reestablished. In this study, we investigated the biological and mechanical properties of an engineered tendon-fibrocartilage-bone composite (TFBC) with cyclic tension (3% strain, 0.2 Hz). Decellularized TFBCs seeded with bone marrow-derived mesenchymal stem cell (BMSCs) sheets and subjected to mechanical stimulation for up to 7 days, were characterized by histology, immunohistochemistry, scanning electron microscopy, mechanical testing, and transcriptional regulation. The decellularized TFBC maintained native enthesis structure and properties. Mechanically stimulated TFBC-BMSC constructs displayed increased cell migration after 7 days of culture compared to static groups. The seeded cell sheet not only integrated well with tendon scaffold but also distributed homogeneously and aligned to the direction of stretch under dynamic culture. Developmental genes were regulated including, scleraxis which was significantly upregulated with mechanical stimulation. The Young's modulus of the cell-seeded constructs was significantly higher compared to the non-cell-seeded controls. In conclusion, the results of this study reveal that the TFBC-BMSC composite provides an ideal multilayer construct for cell seeding and growth, with mechanical preconditioning further enhances cell penetration and differentiation. The BMSC cell sheet revitalized TFBC in conjunction with mechanical stimulation could serve as a novel and primed biological patch to improve rotator cuff repair. This article is protected by copyright. All rights reserved.

  14. Cell dynamics in cervical loop epithelium during transition from crown to root: implications for Hertwig's epithelial root sheath formation.

    PubMed

    Sakano, M; Otsu, K; Fujiwara, N; Fukumoto, S; Yamada, A; Harada, H

    2013-04-01

    Some clinical cases of hypoplastic tooth root are congenital. Because the formation of Hertwig's epithelial root sheath (HERS) is an important event for root development and growth, we have considered that understanding the HERS developmental mechanism contributes to elucidate the causal factors of the disease. To find integrant factors and phenomenon for HERS development and growth, we studied the proliferation and mobility of the cervical loop (CL). We observed the cell movement of CL by the DiI labeling and organ culture system. To examine cell proliferation, we carried out immunostaining of CL and HERS using anti-Ki67 antibody. Cell motility in CL was observed by tooth germ slice organ culture using green fluorescent protein mouse. We also examined the expression of paxillin associated with cell movement. Imaging using DiI labeling showed that, at the apex of CL, the epithelium elongated in tandem with the growth of outer enamel epithelium (OEE). Cell proliferation assay using Ki67 immunostaining showed that OEE divided more actively than inner enamel epithelium (IEE) at the onset of HERS formation. Live imaging suggested that mobility of the OEE and cells in the apex of CL were more active than in IEE. The expression of paxillin was observed strongly in OEE and the apex of CL. The more active growth and movement of OEE cells contributed to HERS formation after reduction of the growth of IEE. The expression pattern of paxillin was involved in the active movement of OEE and HERS. The results will contribute to understand the HERS formation mechanism and elucidate the cause of anomaly root. © 2012 John Wiley & Sons A/S.

  15. Cross-Cultural Adaptation of a Developmental Assessment for Arabic-Speaking Children with Visual Impairment

    ERIC Educational Resources Information Center

    Macrine, Sheila L.; Heji, Hayat; Sabri, Amel; Dalton, Sara

    2015-01-01

    Developmental screening has become an established component of child health programs in many developed countries. The research objective of this project was to translate and adapt a developmental assessment (Oregon Project Skills Inventory) for use with young children with visual impairments who speak Arabic. The study was prompted by the lack of…

  16. Development of REM sleep drive and clinical implications

    PubMed Central

    Kobayashi, T.; Good, C.; Mamiya, K.; Skinner, R.D.; Garcia-Rill, E.

    2015-01-01

    REM sleep in the human declines from about 50% of total sleep time (~8 hours) in the newborn to about 15% of total sleep time (~1 hour) in the adult, and this decrease takes place mainly between birth and the end of puberty. We hypothesize that, if this developmental decrease in REM drive does not occur, lifelong increases in REM sleep drive may ensue. In the rat, the developmental decrease in REM sleep occurs between 10 and 30 days after birth, declining from over 70% of total sleep time in the newborn to the adult level of about 15% of sleep time during this period. Rats aged 12–21 days were anaesthetized with Ketamine, decapitated and brainstem slices cut for intracellular recordings. We found that excitatory responses of pedunculopontine nucleus (PPN) neurons to NMDA decrease, while responses to kainic acid increase, over this critical period. Serotonergic type 1 agonists have increasing inhibitory responses, while serotonergic type 2 agonists do not change, during this developmental period. The results suggest that, as PPN neurons develop, they are increasingly activated by kainic acid and increasingly inhibited by serotonergic type 1 receptors. These processes may be related to the developmental decrease in REM sleep. Developmental disturbances in each of these systems could induce differential increases in REM sleep drive, accounting for the post-pubertal onset of a number of different disorders manifesting increases in REM sleep drive. Examination of modulation by PPN projections to ascending and descending targets revealed the presence of common signals modulating both ascending arousal-related functions and descending postural/locomotor-related functions. PMID:14527968

  17. Developmental change in the contribution of voltage-gated Ca(2+) channels to the pacemaking of deep cerebellar nuclei neurons.

    PubMed

    Alviña, K; Tara, E; Khodakhah, K

    2016-05-13

    The activity of the deep cerebellar nuclei (DCN) neurons conveys the bulk of the output of the cerebellum. To generate these motor signals, DCN neurons integrate synaptic inputs with their own spontaneous activity. We have previously reported that N-type voltage-gated Ca(2+) channels modulate the spontaneous activity of the majority of juvenile DCN neurons in vitro. Specifically, pharmacologically blocking N-type Ca(2+) channels increases their firing rate causing DCN cells to burst. Adult DCN neurons however, behaved differently. To further investigate this change, we have studied here the effect of cadmium on the firing rate of DCN neurons in acute cerebellar slices obtained from adult (>2 months old) or juvenile (12-21 days old) rats and mice. Strikingly, and in contrast to juvenile DCN cells, cadmium did not affect the pacemaking of adult DCN cells. The activity of Purkinje cells (PCs) however was transformed into high-frequency bursting, regardless the age. Further, we questioned whether these findings could be due to an artifact associated with the added difficulty of preparing adult DCN slices. Hence we proceeded to examine the spontaneous activity of DCN neurons in anesthetized juvenile and adult rats and mice in vivo. When cadmium was injected into the DCN in vivo no significant change in firing rate was observed, conversely to most juvenile DCN neurons which showed high-frequency bursts after cadmium injection. In these same animals, PCs pacemaking showed no developmental difference. Thus our results demonstrate a remarkable age-dependent functional modification in the regulation of DCN neurons pacemaking. Copyright © 2016 IBRO. Published by Elsevier Ltd. All rights reserved.

  18. Promoting Cultural Relativism in Counselors through the Cultural De-Centering Model

    ERIC Educational Resources Information Center

    McAuliffe, Garrett J.; Milliken, Tammi F.

    2009-01-01

    Counselors who are culturally encapsulated are likely to create client mistrust and to misinterpret clients' cultural norms. This article presents the Cultural De-Centering Model (CDCM) as a constructive-developmental method for helping future counselors to be less ethnocentric in their work. The goal of the CDCM is to increase counselors'…

  19. Lipopolysaccharide does not alter small airway reactivity in mouse lung slices.

    PubMed

    Donovan, Chantal; Royce, Simon G; Vlahos, Ross; Bourke, Jane E

    2015-01-01

    The bacterial endotoxin, lipopolysaccharide (LPS) has been associated with occupational airway diseases with asthma-like symptoms and in acute exacerbations of COPD. The direct and indirect effects of LPS on small airway reactivity have not been fully elucidated. We tested the hypothesis that both in vitro and in vivo LPS treatment would increase contraction and impair relaxation of mouse small airways. Lung slices were prepared from naïve Balb/C mice and cultured in the absence or presence of LPS (10 μg/ml) for up to 48 h for measurement of TNFα levels in conditioned media. Alternatively, mice were challenged with PBS or LPS in vivo once a day for 4 days for preparation of lung slices or for harvest of lungs for Q-PCR analysis of gene expression of pro-inflammatory cytokines and receptors involved in airway contraction. Reactivity of small airways to contractile agonists, methacholine and serotonin, and bronchodilator agents, salbutamol, isoprenaline and rosiglitazone, were assessed using phase-contrast microscopy. In vitro LPS treatment of slices increased TNFα release 6-fold but did not alter contraction or relaxation to any agonists tested. In vivo LPS treatment increased lung gene expression of TNFα, IL-1β and ryanodine receptor isoform 2 more than 5-fold. However there were no changes in reactivity in lung slices from these mice, even when also incubated with LPS ex vivo. Despite evidence of LPS-induced inflammation, neither airway hyperresponsiveness or impaired dilator reactivity were evident. The increase in ryanodine receptor isoform 2, known to regulate calcium signaling in vascular smooth muscle, warrants investigation. Since LPS failed to elicit changes in small airway reactivity in mouse lung slices following in vitro or in vivo treatment, alternative approaches are required to define the potential contribution of this endotoxin to altered small airway reactivity in human lung diseases.

  20. Lipopolysaccharide Does Not Alter Small Airway Reactivity in Mouse Lung Slices

    PubMed Central

    Donovan, Chantal; Royce, Simon G.; Vlahos, Ross; Bourke, Jane E.

    2015-01-01

    The bacterial endotoxin, lipopolysaccharide (LPS) has been associated with occupational airway diseases with asthma-like symptoms and in acute exacerbations of COPD. The direct and indirect effects of LPS on small airway reactivity have not been fully elucidated. We tested the hypothesis that both in vitro and in vivo LPS treatment would increase contraction and impair relaxation of mouse small airways. Lung slices were prepared from naïve Balb/C mice and cultured in the absence or presence of LPS (10 μg/ml) for up to 48 h for measurement of TNFα levels in conditioned media. Alternatively, mice were challenged with PBS or LPS in vivo once a day for 4 days for preparation of lung slices or for harvest of lungs for Q-PCR analysis of gene expression of pro-inflammatory cytokines and receptors involved in airway contraction. Reactivity of small airways to contractile agonists, methacholine and serotonin, and bronchodilator agents, salbutamol, isoprenaline and rosiglitazone, were assessed using phase-contrast microscopy. In vitro LPS treatment of slices increased TNFα release 6-fold but did not alter contraction or relaxation to any agonists tested. In vivo LPS treatment increased lung gene expression of TNFα, IL-1β and ryanodine receptor isoform 2 more than 5-fold. However there were no changes in reactivity in lung slices from these mice, even when also incubated with LPS ex vivo. Despite evidence of LPS-induced inflammation, neither airway hyperresponsiveness or impaired dilator reactivity were evident. The increase in ryanodine receptor isoform 2, known to regulate calcium signaling in vascular smooth muscle, warrants investigation. Since LPS failed to elicit changes in small airway reactivity in mouse lung slices following in vitro or in vivo treatment, alternative approaches are required to define the potential contribution of this endotoxin to altered small airway reactivity in human lung diseases. PMID:25822969

  1. The Impact of Cultural Diversity on Special Education Provision in the United States

    ERIC Educational Resources Information Center

    Palawat, Manisara; May, Michael E.

    2012-01-01

    Cultural background influences one's understanding of intellectual and/or developmental disabilities. More specifically, the cultural perspectives of parents and special education professionals may affect decision-making in providing appropriate services for children with disabilities. Therefore, cultural distinctions may present a unique…

  2. Cultural and Contextual Issues in Exemplar Research

    ERIC Educational Resources Information Center

    King, Pamela Ebstyne; Oakes Mueller, Ross A.; Furrow, James

    2013-01-01

    This chapter specifically addresses how exemplar methods are especially relevant to examining cultural and contextual issues. Cross-cultural, cultural, and indigenous psychologies are discussed in order to highlight how studying actual exemplars in their unique and complex developmental contexts has the potential to identify themes that either…

  3. Excitatory action of GABA on immature neurons is not due to absence of ketone bodies metabolites or other energy substrates.

    PubMed

    Ben-Ari, Yehezkel; Tyzio, Roman; Nehlig, Astrid

    2011-09-01

    Brain slices incubated with glucose have provided most of our knowledge on cellular, synaptic, and network driven mechanisms. It has been recently suggested that γ-aminobutyric acid (GABA) excites neonatal neurons in conventional glucose-perfused slices but not when ketone bodies metabolites, pyruvate, and/or lactate are added, suggesting that the excitatory actions of GABA are due to energy deprivation when glucose is the sole energy source. In this article, we review the vast number of studies that show that slices are not energy deprived in glucose-containing medium, and that addition of other energy substrates at physiologic concentrations does not alter the excitatory actions of GABA on neonatal neurons. In contrast, lactate, like other weak acids, can produce an intracellular acidification that will cause a reduction of intracellular chloride and a shift of GABA actions. The effects of high concentrations of lactate, and particularly of pyruvate (4-5 mm), as used are relevant primarily to pathologic conditions; these concentrations not being found in the brain in normal "control" conditions. Slices in glucose-containing medium may not be ideal, but additional energy substrates neither correspond to physiologic conditions nor alter GABA actions. In keeping with extensive observations in a wide range of animal species and brain structures, GABA depolarizes immature neurons and the reduction of the intracellular concentration of chloride ([Cl(-)](i)) is a basic property of brain maturation that has been preserved throughout evolution. In addition, this developmental sequence has important clinical implications, notably concerning the higher incidence of seizures early in life and their long-lasting deleterious sequels. Immature neurons have difficulties exporting chloride that accumulates during seizures, leading to permanent increase of [Cl(-)](i) that converts the inhibitory actions of GABA to excitatory and hampers the efficacy of GABA-acting antiepileptic drugs. Wiley Periodicals, Inc. © 2011 International League Against Epilepsy.

  4. The Ontogeny of Cultural Learning.

    PubMed

    Legare, Cristine H; Harris, Paul L

    2016-05-01

    Developmental research has the potential to address some of the critical gaps in our scientific understanding of the role played by cultural learning in ontogenetic outcomes. The goal of this special section was to gather together leading examples of research on cultural learning across a variety of social contexts and caregiving settings. Although the field of developmental psychology continues to struggle with the persistent problem of oversampling U.S. and Western European populations, we argue that the articles in this special section add to the growing evidence that children everywhere draw on a repertoire of cultural learning strategies that optimize their acquisition of the specific practices, beliefs, and values of their communities. We also identify future directions and outline best practices for the conduct of research on cultural learning. © 2016 The Authors. Child Development © 2016 Society for Research in Child Development, Inc.

  5. Roots from distinct plant developmental stages are capable of rapidly selecting their own microbiome without the influence of environmental and soil edaphic factors

    USDA-ARS?s Scientific Manuscript database

    Soil microbes live in close association with plants and are crucial for plant health and fitness. Recent literature revealed that specific microbes were cultured at distinct developmental stages of Arabidopsis. It is not clear how fast the roots, depending on their developmental stage, can alter the...

  6. Cultural values and immigrant entrepreneurship: the Chinese in Singapore.

    PubMed

    Chan, K B; Chiang, S N

    1994-01-01

    "It is the intent of this paper to examine the interrelationships between early socialisation into core Chinese cultural values, international migration and Chinese immigrant entrepreneurship.... It is through a developmental socialisation process by which [cultural] values are articulated in family and kin network dynamics that social organisations begin to develop and define what is popularly understood as the 'Chinese way of doing business'. We argue that among the overseas Chinese, this way of doing business must be viewed historically and developmentally, as it is intimately intertwined with transmigration experiences and their consequences in shaping values necessary for the emergence and development of entrepreneurship." (SUMMARY IN FRE AND SPA) excerpt

  7. Evaluation of culture-based techniques and 454 pyrosequencing for the analysis of fungal diversity in potting media and organic fertilizers.

    PubMed

    Al-Sadi, A M; Al-Mazroui, S S; Phillips, A J L

    2015-08-01

    Potting media and organic fertilizers (OFs) are commonly used in agricultural systems. However, there is a lack of studies on the efficiency of culture-based techniques in assessing the level of fungal diversity in these products. A study was conducted to investigate the efficiency of seven culture-based techniques and pyrosequencing for characterizing fungal diversity in potting media and OFs. Fungal diversity was evaluated using serial dilution, direct plating and baiting with carrot slices, potato slices, radish seeds, cucumber seeds and cucumber cotyledons. Identity of all the isolates was confirmed on the basis of the internal transcribed spacer region of the ribosomal RNA (ITS rRNA) sequence data. The direct plating technique was found to be superior over other culture-based techniques in the number of fungal species detected. It was also found to be simple and the least time consuming technique. Comparing the efficiency of direct plating with 454 pyrosequencing revealed that pyrosequencing detected 12 and 15 times more fungal species from potting media and OFs respectively. Analysis revealed that there were differences between potting media and OFs in the dominant phyla, classes, orders, families, genera and species detected. Zygomycota (52%) and Chytridiomycota (60%) were the predominant phyla in potting media and OFs respectively. The superiority of pyrosequencing over cultural methods could be related to the ability to detect obligate fungi, slow growing fungi and fungi that exist at low population densities. The evaluated methods in this study, especially direct plating and pyrosequencing, may be used as tools to help detect and reduce movement of unwanted fungi between countries and regions. © 2015 The Society for Applied Microbiology.

  8. Two cell circuits of oriented adult hippocampal neurons on self-assembled monolayers for use in the study of neuronal communication in a defined system.

    PubMed

    Edwards, Darin; Stancescu, Maria; Molnar, Peter; Hickman, James J

    2013-08-21

    In this study, we demonstrate the directed formation of small circuits of electrically active, synaptically connected neurons derived from the hippocampus of adult rats through the use of engineered chemically modified culture surfaces that orient the polarity of the neuronal processes. Although synaptogenesis, synaptic communication, synaptic plasticity, and brain disease pathophysiology can be studied using brain slice or dissociated embryonic neuronal culture systems, the complex elements found in neuronal synapses makes specific studies difficult in these random cultures. The study of synaptic transmission in mature adult neurons and factors affecting synaptic transmission are generally studied in organotypic cultures, in brain slices, or in vivo. However, engineered neuronal networks would allow these studies to be performed instead on simple functional neuronal circuits derived from adult brain tissue. Photolithographic patterned self-assembled monolayers (SAMs) were used to create the two-cell "bidirectional polarity" circuit patterns. This pattern consisted of a cell permissive SAM, N-1[3-(trimethoxysilyl)propyl] diethylenetriamine (DETA), and was composed of two 25 μm somal adhesion sites connected with 5 μm lines acting as surface cues for guided axonal and dendritic regeneration. Surrounding the DETA pattern was a background of a non-cell-permissive poly(ethylene glycol) (PEG) SAM. Adult hippocampal neurons were first cultured on coverslips coated with DETA monolayers and were later passaged onto the PEG-DETA bidirectional polarity patterns in serum-free medium. These neurons followed surface cues, attaching and regenerating only along the DETA substrate to form small engineered neuronal circuits. These circuits were stable for more than 21 days in vitro (DIV), during which synaptic connectivity was evaluated using basic electrophysiological methods.

  9. Organotypic cultures as tools for testing neuroactive drugs - link between in-vitro and in-vivo experiments.

    PubMed

    Drexler, B; Hentschke, H; Antkowiak, B; Grasshoff, C

    2010-01-01

    The development of neuroactive drugs is a time consuming procedure. Candidate drugs must be run through a battery of tests, including receptor studies and behavioural tests on animals. As a rule, numerous substances with promising properties as assessed in receptor studies must be eliminated from the development pipeline in advanced test phases because of unforeseen problems like intolerable side-effects or unsatisfactory performance in the whole organism. Clearly, test systems of intermediate complexity would alleviate this inefficiency. In this review, we propose cultured organotypic brain slices as model systems that could bridge the 'interpolation gap' between receptors and the brain, with a focus on the development of new general anaesthetics with lesser side effects. General anaesthesia is based on the modulation of neurotransmitter receptors and other conductances located on neurons in diverse brain regions, including cerebral cortex and spinal cord. It is well known that different components of general anaesthesia, e.g. hypnosis and immobility, are produced by the depression of neuronal activity in distinct brain regions. The ventral horn of the spinal cord is an important structure for the induction of immobility. Thus, the potentially immobilizing effects of a newly designed drug can be estimated from its depressant effect on neuronal network activity in cultured spinal slices. A drug's sedative and hypnotic potential can be examined in cortical cultures. Combined with genetically engineered mice, this approach can point to receptor subtypes most relevant to the drug's intended net effect and in return can help in the design of more selective drugs. In conclusion, the use of organotypic cultures permits predictions of neuroactive properties of newly designed drugs on an intermediate level, and should therefore open up avenues for a more creative and economic drug development process.

  10. The Importance of Culture for Developmental Science

    ERIC Educational Resources Information Center

    Keller, Heidi

    2012-01-01

    In this essay, it is argued that a general understanding of human development needs a unified framework based on evolutionary theorizing and cross-cultural and cultural anthropological approaches. An eco-social model of development has been proposed that defines cultural milieus as adaptations to specific socio-demographic contexts. Ontogenetic…

  11. Mexican-Origin Youth's Cultural Orientations and Adjustment: Changes from Early to Late Adolescence

    ERIC Educational Resources Information Center

    Updegraff, Kimberly A.; Umana-Taylor, Adriana J.; McHale, Susan M.; Wheeler, Lorey A.; Perez-Brena, Norma J.

    2012-01-01

    Drawing from developmental and cultural adaptation perspectives and using a longitudinal design, this study examined: (a) mean-level changes in Mexican-origin adolescents' cultural orientations and adjustment from early to late adolescence and (b) bidirectional associations between cultural orientations and adjustment using a cross-lag panel…

  12. An Introductory Undergraduate Course Covering Animal Cell Culture Techniques

    ERIC Educational Resources Information Center

    Mozdziak, Paul E.; Petitte, James N.; Carson, Susan D.

    2004-01-01

    Animal cell culture is a core laboratory technique in many molecular biology, developmental biology, and biotechnology laboratories. Cell culture is a relatively old technique that has been sparingly taught at the undergraduate level. The traditional methodology for acquiring cell culture training has been through trial and error, instruction when…

  13. Trajectories of Mexican American and Mainstream Cultural Values Among Mexican American Adolescents

    PubMed Central

    Knight, George P.; Basilio, Camille D.; Cham, Heining; Gonzales, Nancy A.; Liu, Yu; Umaña-Taylor, Adriana J.

    2013-01-01

    Mexican Americans are one of the largest and fastest growing ethnic groups in the United States, yet we have limited knowledge regarding changes (i.e., developmental trajectories) in cultural orientation based upon their exposure to the Mexican American and mainstream cultures. We examined the parallel trajectories of Mexican American and mainstream cultural values in a sample of 749 Mexican American adolescents (49% female) across assessments during the fifth grade (approximately 11 years of age), the seventh grade (approximately 13 years of age) and the tenth grade (approximately 16 years of age). We expected that these values would change over this developmental period and this longitudinal approach is more appropriate than the often used median split classification to identify distinct types of acculturation. We found four distinct acculturation trajectory groups: two trajectory groups that were increasing slightly with age in the endorsement of mainstream cultural values, one of which was relatively stable in Mexican American cultural values while the other was declining in their endorsement of these values; and two trajectory groups that were declining substantially with age in their endorsement of mainstream cultural values, one of which was also declining in Mexican American cultural values and the other which was stable in these values. These four trajectory groups differed in expected ways on a number of theoretically related cultural variables, but were not highly consistent with the median split classifications. The findings highlight the need to utilize longitudinal data to examine the developmental changes of Mexican American individual’s adaptation to the ethnic and mainstream culture in order to understand more fully the processes of acculturation and enculturation. PMID:23877194

  14. Trajectories of Mexican American and mainstream cultural values among Mexican American adolescents.

    PubMed

    Knight, George P; Basilio, Camille D; Cham, Heining; Gonzales, Nancy A; Liu, Yu; Umaña-Taylor, Adriana J

    2014-12-01

    Mexican Americans are one of the largest and fastest growing ethnic groups in the United States, yet we have limited knowledge regarding changes (i.e., developmental trajectories) in cultural orientation based upon their exposure to the Mexican American and mainstream cultures. We examined the parallel trajectories of Mexican American and mainstream cultural values in a sample of 749 Mexican American adolescents (49 % female) across assessments during the fifth grade (approximately 11 years of age), the seventh grade (approximately 13 years of age) and the tenth grade (approximately 16 years of age). We expected that these values would change over this developmental period and this longitudinal approach is more appropriate than the often used median split classification to identify distinct types of acculturation. We found four distinct acculturation trajectory groups: two trajectory groups that were increasing slightly with age in the endorsement of mainstream cultural values, one of which was relatively stable in Mexican American cultural values while the other was declining in their endorsement of these values; and two trajectory groups that were declining substantially with age in their endorsement of mainstream cultural values, one of which was also declining in Mexican American cultural values and the other which was stable in these values. These four trajectory groups differed in expected ways on a number of theoretically related cultural variables, but were not highly consistent with the median split classifications. The findings highlight the need to utilize longitudinal data to examine the developmental changes of Mexican American individual's adaptation to the ethnic and mainstream culture in order to understand more fully the processes of acculturation and enculturation.

  15. Survival, migration, and differentiation of Sox1-GFP embryonic stem cells in coculture with an auditory brainstem slice preparation.

    PubMed

    Glavaski-Joksimovic, Aleksandra; Thonabulsombat, Charoensri; Wendt, Malin; Eriksson, Mikael; Palmgren, Björn; Jonsson, Anna; Olivius, Petri

    2008-03-01

    The poor regeneration capability of the mammalian hearing organ has initiated different approaches to enhance its functionality after injury. To evaluate a potential neuronal repair paradigm in the inner ear and cochlear nerve we have previously used embryonic neuronal tissue and stem cells for implantation in vivo and in vitro. At present, we have used in vitro techniques to study the survival and differentiation of Sox1-green fluorescent protein (GFP) mouse embryonic stem (ES) cells as a monoculture or as a coculture with rat auditory brainstem slices. For the coculture, 300 microm-thick brainstem slices encompassing the cochlear nucleus and cochlear nerve were prepared from postnatal SD rats. The slices were propagated using the membrane interface method and the cochlear nuclei were prelabeled with DiI. After some days in culture a suspension of Sox1 cells was deposited next to the brainstem slice. Following deposition Sox1 cells migrated toward the brainstem and onto the cochlear nucleus. GFP was not detectable in undifferentiated ES cells but became evident during neural differentiation. Up to 2 weeks after transplantation the cocultures were fixed. The undifferentiated cells were evaluated with antibodies against progenitor cells whereas the differentiated cells were determined with neuronal and glial markers. The morphological and immunohistochemical data indicated that Sox1 cells in monoculture differentiated into a higher percentage of glial cells than neurons. However, when a coculture was used a significantly lower percentage of Sox1 cells differentiated into glial cells. The results demonstrate that a coculture of Sox1 cells and auditory brainstem present a useful model to study stem cell differentiation.

  16. Subcellular TSC22D4 localization in cerebellum granule neurons of the mouse depends on development and differentiation.

    PubMed

    Canterini, Sonia; Bosco, Adriana; Carletti, Valentina; Fuso, Andrea; Curci, Armando; Mangia, Franco; Fiorenza, Maria Teresa

    2012-03-01

    We previously demonstrated that TSC22D4, a protein encoded by the TGF-β1-activated gene Tsc22d4 (Thg-1pit) and highly expressed in postnatal and adult mouse cerebellum with multiple post-translationally modified protein forms, moves to nucleus when in vitro differentiated cerebellum granule neurons (CGNs) are committed to apoptosis by hyperpolarizing KCl concentrations in the culture medium. We have now studied TSC22D4 cytoplasmic/nuclear localization in CGNs and Purkinje cells: (1) during CGN differentiation/maturation in vivo, (2) during CGN differentiation in vitro, and (3) by in vitro culturing ex vivo cerebellum slices under conditions favoring/inhibiting CGN/Purkinje cell differentiation. We show that TSC22D4 displays both nuclear and cytoplasmic localizations in undifferentiated, early postnatal cerebellum CGNs, irrespectively of CGN proliferation/migration from external to internal granule cell layer, and that it specifically accumulates in the somatodendritic and synaptic compartments when CGNs mature, as indicated by TSC22D4 abundance at the level of adult cerebellum glomeruli and apparent lack in CGN nuclei. These features were also observed in cerebellum slices cultured in vitro under conditions favoring/inhibiting CGN/Purkinje cell differentiation. In vitro TSC22D4 silencing with siRNAs blocked CGN differentiation and inhibited neurite elongation in N1E-115 neuroblastoma cells, pinpointing the relevance of this protein to CGN differentiation.

  17. Blockade of P2X7 receptors or pannexin-1 channels similarly attenuates postischemic damage.

    PubMed

    Cisneros-Mejorado, Abraham; Gottlieb, Miroslav; Cavaliere, Fabio; Magnus, Tim; Koch-Nolte, Friederich; Scemes, Eliana; Pérez-Samartín, Alberto; Matute, Carlos

    2015-05-01

    The role of P2X7 receptors and pannexin-1 channels in ischemic damage remains controversial. Here, we analyzed their contribution to postanoxic depolarization after ischemia in cultured neurons and in brain slices. We observed that pharmacological blockade of P2X7 receptors or pannexin-1 channels delayed the onset of postanoxic currents and reduced their slope, and that simultaneous inhibition did not further enhance the effects of blocking either one. These results were confirmed in acute cortical slices from P2X7 and pannexin-1 knockout mice. Oxygen-glucose deprivation in cortical organotypic cultures caused neuronal death that was reduced with P2X7 and pannexin-1 blockers as well as in organotypic cultures derived from mice lacking P2X7 and pannexin 1. Subsequently, we used transient middle cerebral artery occlusion to monitor the neuroprotective effect of those drugs in vivo. We found that P2X7 and pannexin-1 antagonists, and their ablation in knockout mice, substantially attenuated the motor symptoms and reduced the infarct volume to ~50% of that in vehicle-treated or wild-type animals. These results show that P2X7 receptors and pannexin-1 channels are major mediators of postanoxic depolarization in neurons and of brain damage after ischemia, and that they operate in the same deleterious signaling cascade leading to neuronal and tissue demise.

  18. P2 receptor-stimulation influences axonal outgrowth in the developing hippocampus in vitro.

    PubMed

    Heine, C; Heimrich, B; Vogt, J; Wegner, A; Illes, P; Franke, Heike

    2006-01-01

    Extracellular ATP might act as a trophic factor on growing axons during development of the CNS via P2 receptors. In the present study the postnatal presence of selected P2 receptor subtypes was analyzed and their putative trophic capacity in entorhino-hippocampal slice co-cultures of mouse brain was tested. The effect of the P2 receptor ligands 2-methylthioadenosine-5'-triphosphate (P2X/Y receptor agonist) and pyridoxalphosphate-6-azophenyl-2',4'-disulphonic acid (P2X/Y receptor antagonist) on axonal growth and fiber density of biocytin-labeled hippocampal projections was compared both with untreated cultures and with cultures treated with artificial cerebrospinal fluid. After 10 days in vitro, double immunofluorescence labeling revealed the expression of P2X(1), P2X(2), P2X(4) as well as P2Y(1) and P2Y(2) receptors in the examined regions of entorhinal fiber termination. Further, quantitative analysis of identified biocytin-traced entorhinal fibers showed a significant increase in fiber density in the dentate gyrus after incubation of the slices with the P2 receptor agonist 2-methylthioadenosine-5'-triphosphate. This neurite outgrowth promoting effect was completely abolished by the P2 receptor antagonist pyridoxalphosphate-6-azophenyl-2',4'-disulphonic acid. Our in vitro data indicate that ATP via its P2X and P2Y receptors can shape hippocampal connectivity during development.

  19. Large-scale oscillatory calcium waves in the immature cortex.

    PubMed

    Garaschuk, O; Linn, J; Eilers, J; Konnerth, A

    2000-05-01

    Two-photon imaging of large neuronal networks in cortical slices of newborn rats revealed synchronized oscillations in intracellular Ca2+ concentration. These spontaneous Ca2+ waves usually started in the posterior cortex and propagated slowly (2.1 mm per second) toward its anterior end. Ca2+ waves were associated with field-potential changes and required activation of AMPA and NMDA receptors. Although GABAA receptors were not involved in wave initiation, the developmental transition of GABAergic transmission from depolarizing to hyperpolarizing (around postnatal day 7) stopped the oscillatory activity. Thus we identified a type of large-scale Ca2+ wave that may regulate long-distance wiring in the immature cortex.

  20. Culture-Shock and Reverse-Culture Shock: Implications for Juniors Abroad and Seniors at Home.

    ERIC Educational Resources Information Center

    Hogan, John T.

    Thousands of college seniors who have returned from their junior year abroad may be enduring "reverse culture shock" or "reentry crisis." Social psychology and sociology, in the form of "sojourn research," has derived a developmental, stage specific model of culture shock and reverse culture shock, similar to the grieving process identified by…

  1. Mastering Developmental Transitions in Immigrant Adolescents: The Longitudinal Interplay of Family Functioning, Developmental and Acculturative Tasks

    ERIC Educational Resources Information Center

    Reitz, Anne K.; Motti-Stefanidi, Frosso; Asendorpf, Jens B.

    2014-01-01

    Immigrant youth differ in their adaptation, which is judged on the basis of how well they deal with developmental and acculturative tasks. While immigrant adolescents are faced with the realities of 2 different cultures, they also have to master age-salient tasks, such as self-efficacy and identity development. To get a better insight into the…

  2. Entorhinal Denervation Induces Homeostatic Synaptic Scaling of Excitatory Postsynapses of Dentate Granule Cells in Mouse Organotypic Slice Cultures

    PubMed Central

    Vlachos, Andreas; Becker, Denise; Jedlicka, Peter; Winkels, Raphael; Roeper, Jochen; Deller, Thomas

    2012-01-01

    Denervation-induced changes in excitatory synaptic strength were studied following entorhinal deafferentation of hippocampal granule cells in mature (≥3 weeks old) mouse organotypic entorhino-hippocampal slice cultures. Whole-cell patch-clamp recordings revealed an increase in excitatory synaptic strength in response to denervation during the first week after denervation. By the end of the second week synaptic strength had returned to baseline. Because these adaptations occurred in response to the loss of excitatory afferents, they appeared to be in line with a homeostatic adjustment of excitatory synaptic strength. To test whether denervation-induced changes in synaptic strength exploit similar mechanisms as homeostatic synaptic scaling following pharmacological activity blockade, we treated denervated cultures at 2 days post lesion for 2 days with tetrodotoxin. In these cultures, the effects of denervation and activity blockade were not additive, suggesting that similar mechanisms are involved. Finally, we investigated whether entorhinal denervation, which removes afferents from the distal dendrites of granule cells while leaving the associational afferents to the proximal dendrites of granule cells intact, results in a global or a local up-scaling of granule cell synapses. By using computational modeling and local electrical stimulations in Strontium (Sr2+)-containing bath solution, we found evidence for a lamina-specific increase in excitatory synaptic strength in the denervated outer molecular layer at 3–4 days post lesion. Taken together, our data show that entorhinal denervation results in homeostatic functional changes of excitatory postsynapses of denervated dentate granule cells in vitro. PMID:22403720

  3. The role of Nedd4-1 WW domains in binding and regulating human organic anion transporter 1.

    PubMed

    Xu, Da; Wang, Haoxun; Gardner, Carol; Pan, Zui; Zhang, Ping L; Zhang, Jinghui; You, Guofeng

    2016-08-01

    Human organic anion transporter 1 (hOAT1), expressed at the basolateral membrane of kidney proximal tubule cells, mediates the active renal secretion of a diverse array of clinically important drugs, including anti-human immunodeficiency virus therapeutics, antitumor drugs, antibiotics, antihypertensives, and anti-inflammatories. We have previously demonstrated that posttranslational modification of hOAT1 by ubiquitination is an important mechanism for the regulation of this transporter. The present study aimed at identifying the ubiquitin ligase for hOAT1 and its mechanism of action. We showed that overexpression of neural precursor cell expressed, developmentally downregulated (Nedd)4-1, an E3 ubiquitin ligase, enhanced hOAT1 ubiquitination, decreased hOAT1 expression at the cell surface, and inhibited hOAT1 transport activity. In contrast, overexpression of the ubiquitin ligase-dead mutant Nedd4-1/C867S was without effects on hOAT1. Furthermore, knockdown of endogenously expressed Nedd4-1 by Nedd4-1-specific small interfering RNA reduced hOAT1 ubiquitination. Immunoprecipitation experiments in cultured cells and rat kidney slices and immunofluorescence experiments in rat kidney slices showed that there was a physical interaction between OAT1 and Nedd4-1. Nedd4-1 contains four protein-protein interacting WW domains. When these WW domains were inactivated by mutating two amino acid residues in each of the four WW domains (Mut-WW1: V210W/H212G, Mut-WW2: V367W/H369G, Mut-WW3: I440W/H442G, and Mut-WW4: I492W/H494G, respectively), only Mut-WW2 and Mut-WW3 significantly lost their ability to bind and to ubiquitinate hOAT1. As a result, Mut-WW2 and Mut-WW3 were unable to suppress hOAT1-mediated transport as effectively as wild-type Nedd4-1. In conclusion, this is the first demonstration that Nedd4-1 regulates hOAT1 ubiquitination, expression, and transport activity through its WW2 and WW3 domains. Copyright © 2016 the American Physiological Society.

  4. The role of Nedd4-1 WW domains in binding and regulating human organic anion transporter 1

    PubMed Central

    Xu, Da; Wang, Haoxun; Gardner, Carol; Pan, Zui; Zhang, Ping L.; Zhang, Jinghui

    2016-01-01

    Human organic anion transporter 1 (hOAT1), expressed at the basolateral membrane of kidney proximal tubule cells, mediates the active renal secretion of a diverse array of clinically important drugs, including anti-human immunodeficiency virus therapeutics, antitumor drugs, antibiotics, antihypertensives, and anti-inflammatories. We have previously demonstrated that posttranslational modification of hOAT1 by ubiquitination is an important mechanism for the regulation of this transporter. The present study aimed at identifying the ubiquitin ligase for hOAT1 and its mechanism of action. We showed that overexpression of neural precursor cell expressed, developmentally downregulated (Nedd)4-1, an E3 ubiquitin ligase, enhanced hOAT1 ubiquitination, decreased hOAT1 expression at the cell surface, and inhibited hOAT1 transport activity. In contrast, overexpression of the ubiquitin ligase-dead mutant Nedd4-1/C867S was without effects on hOAT1. Furthermore, knockdown of endogenously expressed Nedd4-1 by Nedd4-1-specific small interfering RNA reduced hOAT1 ubiquitination. Immunoprecipitation experiments in cultured cells and rat kidney slices and immunofluorescence experiments in rat kidney slices showed that there was a physical interaction between OAT1 and Nedd4-1. Nedd4-1 contains four protein-protein interacting WW domains. When these WW domains were inactivated by mutating two amino acid residues in each of the four WW domains (Mut-WW1: V210W/H212G, Mut-WW2: V367W/H369G, Mut-WW3: I440W/H442G, and Mut-WW4: I492W/H494G, respectively), only Mut-WW2 and Mut-WW3 significantly lost their ability to bind and to ubiquitinate hOAT1. As a result, Mut-WW2 and Mut-WW3 were unable to suppress hOAT1-mediated transport as effectively as wild-type Nedd4-1. In conclusion, this is the first demonstration that Nedd4-1 regulates hOAT1 ubiquitination, expression, and transport activity through its WW2 and WW3 domains. PMID:27226107

  5. Depression risks in mothers of children with developmental disabilities: a cross-cultural comparison of Brazil, Colombia, Malaysia and Thailand.

    PubMed

    Osada, Hirokazu; Coelho de Amorim, Annibal; Velosa, Andrea; Wan, Wong Poh; Lotrakul, Panpimol; Hara, Hitoshi

    2013-06-01

    Compared with US or European countries, there are fewer mental health services for mothers of children with developmental disabilities in Latin American and/or Southeast Asian countries. To explore the risk of depression in mothers of children with developmental disabilities in countries with a lack of mental health professionals, we conducted cross-cultural comparisons for four countries: Brazil, Colombia, Malaysia and Thailand. Using the CES-D, we compared the participants' depressive symptoms, by which we also estimated the probability of morbid depression. In every country, participants tended to show depressive symptoms. In the CES-D total scores and the numbers of mothers who were observed to have a high level of depressive symptoms, there were significant differences among countries (F = 4.36, p = .006; χ2 = 10.3, p = .015). Considering cultural models, we could apply evidence-based intervention to depressive mothers of children, and conduct intervention and treatment for those mothers and evaluate ways of providing better mental health services to these individuals.

  6. A Developmental Model of Cross-Cultural Competence at the Tactical Level

    DTIC Science & Technology

    2010-11-01

    components of 3C and describe how 3C develops in Soldiers. Five components of 3C were identified: Cultural Maturity , Cognitive Flexibility, Cultural...a result of the data analysis: Cultural Maturity , Cognitive Flexibility, Cultural Knowledge, Cultural Acuity, and Interpersonal Skills. These five...create regressions in the 3C development process. In short, KSAAs mature interdependently and simultaneously. Thus, development and transitions across

  7. Endogenous neurotrophin-3 regulates short-term plasticity at lateral perforant path-granule cell synapses.

    PubMed

    Kokaia, M; Asztely, F; Olofsdotter, K; Sindreu, C B; Kullmann, D M; Lindvall, O

    1998-11-01

    In the adult brain, neurotrophin-3 (NT-3) is mainly localized in dentate granule cells, and its expression is decreased by various stimuli, e.g., seizure activity. We have examined the role of endogenous NT-3 for excitatory synaptic transmission at lateral perforant path-dentate granule cell synapses using hippocampal slices from NT-3 knock-out (+/-) and wild-type (+/+) mice. Paired-pulse facilitation (PPF) and also short-term synaptic plasticity induced by a brief, high-frequency train of afferent stimulation were reduced, but the expression of long-term potentiation was not affected in the NT-3+/- mice. Incubation of the slices with recombinant NT-3 reversed the deficit in PPF through a mechanism requiring de novo protein synthesis, implying that the impaired short-term plasticity does not result from a developmental alteration. No changes of overall presynaptic release probability, measured by the progressive block of NMDA receptor-mediated synaptic currents by MK-801, or desensitization of AMPA receptors were detected. Because NT-3 expression is reduced after focal seizures, impaired short-term facilitation may represent a protective response that limits the propagation of epileptiform activity from the entorhinal cortex to the hippocampus.

  8. Screening for Developmental Neurotoxicants using In Vitro "Brain on a Chip" Cultures

    EPA Science Inventory

    Currently there are thousands of chemicals in the environment that have not been screened for their potential to cause developmental neurotoxicity (DNT). The use of microelectrode array (MEA) technology allows for simultaneous extracellular measurement of action potential (spike)...

  9. The symphony of the damned: racial discourse, complex political emergencies and humanitarian aid.

    PubMed

    Duffield, M

    1996-09-01

    This paper concerns the manner in which the West is responding to protracted political crises beyond its borders. It examines the conceptual world-view that aid agencies bring to complex emergencies and which shapes action. The paper provides an analysis of developmentalism. That is, the currently dominant idea of development which is an adapted form of multiculturalism. It is based on the empowerment of cultural differences and the relativisation of progress. As a variant of multiculturalism, developmentalism is part of Western racial discourse. In terms of understanding conflict, it establishes a mirror-image relationship with new rascist ideas premised on cultural pluralism inevitably leading to social breakdown, violence and anarchy. To the contrary, with its functional view of social harmony, libertine developmentalism claims that even unresolved political crisis constitutes a development opportunity. Developmentalism, like culturalism generally, is incapable of analysing power. It therefore cannot understand the effects and significance of its own organisational forms. Moreover, since the absence of power translates into operational neutrality in a war zone, it is also unable to analyse the nature of new political formations emerging in the global periphery. That is, the so-called weak or failed states, warlords and so on. This functional ignorance has allowed a widespread incorporation of humanitarian aid into the fabric of political violence. Developmentalism is an essential underpinning for the growing organisational accommodation to ongoing conflict and eroding standards of justice and accountability.

  10. Tumor Slice Culture: A New Avatar in Personalized Oncology

    DTIC Science & Technology

    2017-08-01

    official Department of the Army position, policy or decision unless so designated by other documentation. REPORT DOCUMENTATION PAGE Form Approved OMB No...sensitivity and to correlate the results with clinical and molecular data. 15. SUBJECT TERMS 16. SECURITY CLASSIFICATION OF: 17. LIMITATION OF ABSTRACT...differences in pre-operative treatments. Indeed, the viability scores significantly correlated with pathologic assessment of tumor viability/necrosis

  11. Cultural Artifacts as Scaffolds for Genre Development.

    ERIC Educational Resources Information Center

    Kamberelis, George; Bovino, Thomas D.

    1999-01-01

    Shows that children in the primary grades possessed considerable working knowledge of the cultural conventions of narrative genres but much less working knowledge of the cultural conventions of informational genres. Reveals grade-related developmental differences for some dimensions of linguistic and textual organization. Shows that cultural…

  12. Counseling Third Culture Kids.

    ERIC Educational Resources Information Center

    Barringer, Carolyn Fox

    Third Culture Kids (TCKs) represent a group of youth who have lived overseas with their families for business, service, or missionary work. The implications of living in multiple cultures, especially during the developmental and formative years of youth, warrant investigation. This study informs the US counseling community about the…

  13. Radicinols and radicinin phytotoxins produced by Alternaria radicina on carrots.

    PubMed

    Solfrizzo, Michele; Vitti, Carolina; De Girolamo, Annalisa; Visconti, Angelo; Logrieco, Antonio; Fanizzi, Francesco P

    2004-06-02

    The phytotoxin epi-radicinol, a diastereomer of radicinol, was isolated from large cultures of Alternaria radicina grown on carrot slices and identified by GC-MS, LC-MS, (1)H NMR, and (13)C NMR. Four strains of A. radicina isolated from rotted carrot produced epi-radicinol as the major metabolite (up to 39414 microg/g) together with radicinol (up to 2423 microg/g), and, to a lesser extent, radicinin when cultured on carrot slices, whereas on rice they mainly produced radicinin (2486-53800 microg/g). Radicinin and epi-radicinol reduced root elongation of germinating carrot seeds at concentrations of 10-20 microg/mL. Carrot samples naturally infected by A. radicina contained detectable quantities of epi-radicinol also in combination with lower levels of radicinin or radicinol. Accumulation of radicinols and radicinin in stored carrots, either naturally contaminated or artificially inoculated with A. radicina, was stimulated by successive temperature rises from 1 to 10 degrees C and from 10 to 20 degrees C, reaching maximum levels of 60 microg/g epi-radicinol and 26 microg/g radicinin. This is the first report on the production of radicinols by A. radicina and its natural occurrence in carrots in association with radicinin.

  14. Electroosmotic Sampling. Application to Determination of Ectopeptidase Activity in Organotypic Hippocampal Slice Cultures

    PubMed Central

    Xu, Hongjuan; Guy, Yifat; Hamsher, Amy; Shi, Guoyue; Sandberg, Mats; Weber, Stephen G.

    2010-01-01

    We hypothesize that peptide-containing solutions pulled through tissue should reveal the presence and activity of peptidases in the tissue. Using the natural ζ-potential in the organotypic hippocampal slice culture (OHSC), physiological fluids can be pulled through the tissue with an electric field. The hydrolysis of the peptides present in the fluid drawn through the tissue can be determined using capillary HPLC with electrochemical detection of the biuret complexes of the peptides following a postcolumn reaction. We have characterized this new sampling method by measuring the flow rate, examining the use of internal standards, and examining cell death caused by sampling. The sampling flow rate ranges from 60 to 150 nL/min with a 150 μm (ID) sampling capillary with an electric field (at the tip of the capillary) from 30 to 60 V/cm. Cell death can be negligible with controlled sampling conditions. Using this sampling approach, we have electroosmotically pulled Leu-enkephalin through OHSCs to identify ectopeptidase activity in the CA3 region. These studies show that a bestatin-sensitive aminopeptidase may be critical for the hydrolysis of exogenous Leu-enkephalin, a neuropeptide present in the CA3 region of OHSCs. PMID:20669992

  15. A study of cryogenic tissue-engineered liver slices in calcium alginate gel for drug testing.

    PubMed

    Chen, Ruomeng; Wang, Bo; Liu, Yaxiong; Lin, Rong; He, Jiankang; Li, Dichen

    2018-06-01

    To address issues such as transportation and the time-consuming nature of tissue-engineered liver for use as an effective drug metabolism and toxicity testing model, "ready-to-use" cryogenic tissue-engineered liver needs to be studied. The research developed a cryogenic tissue-engineered liver slice (TELS), which comprised of HepG2 cells and calcium alginate gel. Cell viability and liver-specific functions were examined after different cryopreservation and recovery culture times. Then, cryogenic TELSs were used as a drug-testing model and treated with Gefitinib. Cryogenic TELSs were stored at -80 °C to ensure high cell viability. During recovery in culture, the cells in the cryogenic TELS were evenly distributed, massively proliferated, and then formed spheroid-like aggregates from day 1 to day 13. The liver-specific functions in the cryogenic TELS were closely related to cryopreservation time and cell proliferation. As a reproducible drug-testing model, the cryogenic TELS showed an obvious drug reaction after treatment with the Gefitinib. The present study shows that the cryopreservation techniques can be used in drug-testing models. Copyright © 2018 Elsevier Inc. All rights reserved.

  16. Polythiophenes Inhibit Prion Propagation by Stabilizing Prion Protein (PrP) Aggregates*

    PubMed Central

    Margalith, Ilan; Suter, Carlo; Ballmer, Boris; Schwarz, Petra; Tiberi, Cinzia; Sonati, Tiziana; Falsig, Jeppe; Nyström, Sofie; Hammarström, Per; Åslund, Andreas; Nilsson, K. Peter R.; Yam, Alice; Whitters, Eric; Hornemann, Simone; Aguzzi, Adriano

    2012-01-01

    Luminescent conjugated polymers (LCPs) interact with ordered protein aggregates and sensitively detect amyloids of many different proteins, suggesting that they may possess antiprion properties. Here, we show that a variety of anionic, cationic, and zwitterionic LCPs reduced the infectivity of prion-containing brain homogenates and of prion-infected cerebellar organotypic cultured slices and decreased the amount of scrapie isoform of PrPC (PrPSc) oligomers that could be captured in an avidity assay. Paradoxically, treatment enhanced the resistance of PrPSc to proteolysis, triggered the compaction, and enhanced the resistance to proteolysis of recombinant mouse PrP(23–231) fibers. These results suggest that LCPs act as antiprion agents by transitioning PrP aggregates into structures with reduced frangibility. Moreover, ELISA on cerebellar organotypic cultured slices and in vitro conversion assays with mouse PrP(23–231) indicated that poly(thiophene-3-acetic acid) may additionally interfere with the generation of PrPSc by stabilizing the conformation of PrPC or of a transition intermediate. Therefore, LCPs represent a novel class of antiprion agents whose mode of action appears to rely on hyperstabilization, rather than destabilization, of PrPSc deposits. PMID:22493452

  17. Microbiota of regular sodium and sodium-reduced ready-to-eat meat products obtained from the retail market.

    PubMed

    Miller, Petr; Liu, Xiaoji; McMullen, Lynn M

    2015-02-01

    The aim of this study was to assess the influence of sodium content on the microbiota on the surface of ready-to-eat (RTE) meat products purchased from the retail market in Canada. Products, including sliced and sausage-type deli meats, were analysed with culture-dependent and culture-independent methods. Bacteria were identified from 23 brands of products from different meat processors with claims of sodium content ranging from 390 to 1200 mg per 100 g of product. Out of 150 bacterial isolates, the most common were identified as Leuconostoc gelidum, Carnobacterium maltaromaticum, Brochothrix thermosphacta, and Leuconostoc gasicomitatum. Vacuum-packaged RTE deli sliced meat products had the largest population of bacteria. Leuconostocci were the most common isolates in this group of products, while carnobacteria were prevalent on products with moderate loads of bacteria. A higher incidence of carnobacteria and lower incidence of B. thermosphacta were detected on sodium-reduced products. Simpson's and Shannon-Wiener indices showed that low sodium products (25%-50% less sodium) had an overall higher bacterial diversity. This was also observed when individual low sodium products were compared with their regular sodium counterpart.

  18. Characterization of the in vitro propagation of epileptiform electrophysiological activity in organotypic hippocampal slice cultures coupled to 3D microelectrode arrays.

    PubMed

    Pisciotta, Marzia; Morgavi, Giovanna; Jahnsen, Henrik

    2010-10-28

    Dynamic aspects of the propagation of epileptiform activity have so far received little attention. With the aim of providing new insights about the spatial features of the propagation of epileptic seizures in the nervous system, we studied in vitro the initiation and propagation of traveling epileptiform waves of electrophysiological activity in the hippocampus by means of substrate three-dimensional microelectrode arrays (MEAs) for extracellular measurements. Pharmacologically disinhibited hippocampal slices spontaneously generate epileptiform bursts mostly originating in CA3 and propagating to CA1. Our study specifically addressed the activity-dependent changes of the propagation of traveling electrophysiological waves in organotypic hippocampal slices during epileptiform discharge and in particular our question is: what happens to the epileptic signals during their propagation through the slice? Multichannel data analysis enabled us to quantify an activity-dependent increase in the propagation velocity of spontaneous bursts. Moreover, through the evaluation of the coherence of the signals, it was possible to point out that only the lower-frequency components (<95Hz) of the electrical activity are completely coherent with respect to the activity originating in the CA3, while components at higher frequencies lose the coherence, possibly suggesting that the cellular mechanism mediating propagation of electrophysiological activity becomes ineffective for those firing rates exceeding an upper bound or that some noise of neuronal origin was added to the signal during propagation. Copyright © 2010 Elsevier B.V. All rights reserved.

  19. Glucose regulates hypothalamic long-chain fatty acid metabolism via AMP-activated kinase (AMPK) in neurons and astrocytes.

    PubMed

    Taïb, Bouchra; Bouyakdan, Khalil; Hryhorczuk, Cécile; Rodaros, Demetra; Fulton, Stephanie; Alquier, Thierry

    2013-12-27

    Hypothalamic controls of energy balance rely on the detection of circulating nutrients such as glucose and long-chain fatty acids (LCFA) by the mediobasal hypothalamus (MBH). LCFA metabolism in the MBH plays a key role in the control of food intake and glucose homeostasis, yet it is not known if glucose regulates LCFA oxidation and esterification in the MBH and, if so, which hypothalamic cell type(s) and intracellular signaling mechanisms are involved. The aim of this study was to determine the impact of glucose on LCFA metabolism, assess the role of AMP-activated Kinase (AMPK), and to establish if changes in LCFA metabolism and its regulation by glucose vary as a function of the kind of LCFA, cell type, and brain region. We show that glucose inhibits palmitate oxidation via AMPK in hypothalamic neuronal cell lines, primary hypothalamic astrocyte cultures, and MBH slices ex vivo but not in cortical astrocytes and slice preparations. In contrast, oleate oxidation was not affected by glucose or AMPK inhibition in MBH slices. In addition, our results show that glucose increases palmitate, but not oleate, esterification into neutral lipids in neurons and MBH slices but not in hypothalamic astrocytes. These findings reveal for the first time the metabolic fate of different LCFA in the MBH, demonstrate AMPK-dependent glucose regulation of LCFA oxidation in both astrocytes and neurons, and establish metabolic coupling of glucose and LCFA as a distinguishing feature of hypothalamic nuclei critical for the control of energy balance.

  20. Live imaging of mitosis in the developing mouse embryonic cortex.

    PubMed

    Pilaz, Louis-Jan; Silver, Debra L

    2014-06-04

    Although of short duration, mitosis is a complex and dynamic multi-step process fundamental for development of organs including the brain. In the developing cerebral cortex, abnormal mitosis of neural progenitors can cause defects in brain size and function. Hence, there is a critical need for tools to understand the mechanisms of neural progenitor mitosis. Cortical development in rodents is an outstanding model for studying this process. Neural progenitor mitosis is commonly examined in fixed brain sections. This protocol will describe in detail an approach for live imaging of mitosis in ex vivo embryonic brain slices. We will describe the critical steps for this procedure, which include: brain extraction, brain embedding, vibratome sectioning of brain slices, staining and culturing of slices, and time-lapse imaging. We will then demonstrate and describe in detail how to perform post-acquisition analysis of mitosis. We include representative results from this assay using the vital dye Syto11, transgenic mice (histone H2B-EGFP and centrin-EGFP), and in utero electroporation (mCherry-α-tubulin). We will discuss how this procedure can be best optimized and how it can be modified for study of genetic regulation of mitosis. Live imaging of mitosis in brain slices is a flexible approach to assess the impact of age, anatomy, and genetic perturbation in a controlled environment, and to generate a large amount of data with high temporal and spatial resolution. Hence this protocol will complement existing tools for analysis of neural progenitor mitosis.

  1. In vivo Postnatal Electroporation and Time-lapse Imaging of Neuroblast Migration in Mouse Acute Brain Slices

    PubMed Central

    Oudin, Madeleine Julie; Doherty, Patrick; Lalli, Giovanna

    2013-01-01

    The subventricular zone (SVZ) is one of the main neurogenic niches in the postnatal brain. Here, neural progenitors proliferate and give rise to neuroblasts able to move along the rostral migratory stream (RMS) towards the olfactory bulb (OB). This long-distance migration is required for the subsequent maturation of newborn neurons in the OB, but the molecular mechanisms regulating this process are still unclear. Investigating the signaling pathways controlling neuroblast motility may not only help understand a fundamental step in neurogenesis, but also have therapeutic regenerative potential, given the ability of these neuroblasts to target brain sites affected by injury, stroke, or degeneration. In this manuscript we describe a detailed protocol for in vivo postnatal electroporation and subsequent time-lapse imaging of neuroblast migration in the mouse RMS. Postnatal electroporation can efficiently transfect SVZ progenitor cells, which in turn generate neuroblasts migrating along the RMS. Using confocal spinning disk time-lapse microscopy on acute brain slice cultures, neuroblast migration can be monitored in an environment closely resembling the in vivo condition. Moreover, neuroblast motility can be tracked and quantitatively analyzed. As an example, we describe how to use in vivo postnatal electroporation of a GFP-expressing plasmid to label and visualize neuroblasts migrating along the RMS. Electroporation of shRNA or CRE recombinase-expressing plasmids in conditional knockout mice employing the LoxP system can also be used to target genes of interest. Pharmacological manipulation of acute brain slice cultures can be performed to investigate the role of different signaling molecules in neuroblast migration. By coupling in vivo electroporation with time-lapse imaging, we hope to understand the molecular mechanisms controlling neuroblast motility and contribute to the development of novel approaches to promote brain repair. PMID:24326479

  2. Regulation of Brain-Derived Neurotrophic Factor Exocytosis and Gamma-Aminobutyric Acidergic Interneuron Synapse by the Schizophrenia Susceptibility Gene Dysbindin-1.

    PubMed

    Yuan, Qiang; Yang, Feng; Xiao, Yixin; Tan, Shawn; Husain, Nilofer; Ren, Ming; Hu, Zhonghua; Martinowich, Keri; Ng, Julia S; Kim, Paul J; Han, Weiping; Nagata, Koh-Ichi; Weinberger, Daniel R; Je, H Shawn

    2016-08-15

    Genetic variations in dystrobrevin binding protein 1 (DTNBP1 or dysbindin-1) have been implicated as risk factors in the pathogenesis of schizophrenia. The encoded protein dysbindin-1 functions in the regulation of synaptic activity and synapse development. Intriguingly, a loss of function mutation in Dtnbp1 in mice disrupted both glutamatergic and gamma-aminobutyric acidergic transmission in the cerebral cortex; pyramidal neurons displayed enhanced excitability due to reductions in inhibitory synaptic inputs. However, the mechanism by which reduced dysbindin-1 activity causes inhibitory synaptic deficits remains unknown. We investigated the role of dysbindin-1 in the exocytosis of brain-derived neurotrophic factor (BDNF) from cortical excitatory neurons, organotypic brain slices, and acute slices from dysbindin-1 mutant mice and determined how this change in BDNF exocytosis transsynaptically affected the number of inhibitory synapses formed on excitatory neurons via whole-cell recordings, immunohistochemistry, and live-cell imaging using total internal reflection fluorescence microscopy. A decrease in dysbindin-1 reduces the exocytosis of BDNF from cortical excitatory neurons, and this reduction in BDNF exocytosis transsynaptically resulted in reduced inhibitory synapse numbers formed on excitatory neurons. Furthermore, application of exogenous BDNF rescued the inhibitory synaptic deficits caused by the reduced dysbindin-1 level in both cultured cortical neurons and slice cultures. Taken together, our results demonstrate that these two genes linked to risk for schizophrenia (BDNF and dysbindin-1) function together to regulate interneuron development and cortical network activity. This evidence supports the investigation of the association between dysbindin-1 and BDNF in humans with schizophrenia. Copyright © 2016 Society of Biological Psychiatry. Published by Elsevier Inc. All rights reserved.

  3. Prolonged Mek1/2 suppression impairs the developmental potential of embryonic stem cells

    PubMed Central

    Choi, Jiho; Huebner, Aaron J; Clement, Kendell; Walsh, Ryan M; Savol, Andrej; Lin, Kaixuan; Gu, Hongcang; Di Stefano, Bruno; Brumbaugh, Justin; Kim, Sang-Yong; Sharif, Jafar; Rose, Christopher M.; Mohammad, Arman; Odajima, Junko; Charron, Jean; Shioda, Toshi; Gnirke, Andreas; Gygi, Steven; Koseki, Haruhiko; Sadreyev, Ruslan I.; Xiao, Andrew; Meissner, Alexander; Hochedlinger, Konrad

    2018-01-01

    Concomitant activation of the Wnt pathway and suppression of Mapk signaling by two small molecules in the presence of LIF (2i/L) induces a naïve state in mouse embryonic stem cells (ESCs) that resembles the inner cell mass (ICM) of the pre-implantation embryo1. Since the ICM exists only transiently in vivo, it remains unclear how sustained propagation of naïve ESCs in vitro affects their stability and functionality. Here we show that prolonged culture of male ESCs in 2i/L results in irreversible epigenetic and genomic changes that impair their developmental potential. Additionally, we find that female ESCs cultured in conventional serum/LIF (S/L) media phenocopy male ESCs cultured in 2i/L. Mechanistically, we demonstrate that Mek1/2 inhibition is predominantly responsible for these effects, in part through downregulation of DNA methyltransferases and their associated cofactors. Finally, we show that replacement of the Mek1/2 inhibitor with a Src inhibitor preserves the epigenetic and genomic integrity as well as developmental potential of ESCs. Taken together, our data suggest that, while short-term suppression of Mek1/2 in ESCs helps maintain an ICM-like epigenetic state, prolonged suppression results in irreversible changes that compromise their developmental potential. PMID:28746311

  4. Psychopathology, adversity, and creativity: diversifying experiences in the development of eminent African Americans.

    PubMed

    Damian, Rodica Ioana; Simonton, Dean Keith

    2015-04-01

    Symptoms associated with mental illness have been hypothesized to relate to creative achievement because they act as diversifying experiences. However, this theory has only been tested on predominantly majority-culture samples. Do tendencies toward mental illness still predict eminent creativity when they coexist with other diversifying experiences, such as early parental death, minority-status, or poverty? These alternative diversifying experiences can be collectively referred to as examples of developmental adversity. This conjecture was tested on a significant sample of 291 eminent African Americans who, by the nature of their status as long-term minorities, would experience more developmental adversity. Replicating majority-culture patterns, African American artists showed higher mental illness rates than African American scientists. Yet the absolute percentages were significantly lower for the African Americans, regardless of profession. Furthermore, mental illness predicted higher eminence levels only for the African American artists, an effect that diminished when controlling for developmental adversity. Because the latter predicted eminence for both artists and scientists, the "madness-to-genius" link probably represents just 1 of several routes by which diversifying experiences can influence eminence. The same developmental ends can be attained by different means. This inference warrants further research using other eminent creators emerging from minority culture populations. (c) 2015 APA, all rights reserved).

  5. A Simplified Method for Three-Dimensional (3-D) Ovarian Tissue Culture Yielding Oocytes Competent to Produce Full-Term Offspring in Mice

    PubMed Central

    Higuchi, Carolyn M.; Maeda, Yuuki; Horiuchi, Toshitaka; Yamazaki, Yukiko

    2015-01-01

    In vitro growth of follicles is a promising technology to generate large quantities of competent oocytes from immature follicles and could expand the potential of assisted reproductive technologies (ART). Isolated follicle culture is currently the primary method used to develop and mature follicles in vitro. However, this procedure typically requires complicated, time-consuming procedures, as well as destruction of the normal ovarian microenvironment. Here we describe a simplified 3-D ovarian culture system that can be used to mature multilayered secondary follicles into antral follicles, generating developmentally competent oocytes in vitro. Ovaries recovered from mice at 14 days of age were cut into 8 pieces and placed onto a thick Matrigel drop (3-D culture) for 10 days of culture. As a control, ovarian pieces were cultured on a membrane filter without any Matrigel drop (Membrane culture). We also evaluated the effect of activin A treatment on follicle growth within the ovarian pieces with or without Matrigel support. Thus we tested four different culture conditions: C (Membrane/activin-), A (Membrane/activin+), M (Matrigel/activin-), and M+A (Matrigel/activin+). We found that the cultured follicles and oocytes steadily increased in size regardless of the culture condition used. However, antral cavity formation occurred only in the follicles grown in the 3-D culture system (M, M+A). Following ovarian tissue culture, full-grown GV oocytes were isolated from the larger follicles to evaluate their developmental competence by subjecting them to in vitro maturation (IVM) and in vitro fertilization (IVF). Maturation and fertilization rates were higher using oocytes grown in 3-D culture (M, M+A) than with those grown in membrane culture (C, A). In particular, activin A treatment further improved 3-D culture (M+A) success. Following IVF, two-cell embryos were transferred to recipients to generate full-term offspring. In summary, this simple and easy 3-D ovarian culture system using a Matrigel drop and activin A supplementation (M+A) provides optimal and convenient conditions to support growth of developmentally competent oocytes in vitro. PMID:26571501

  6. Time-lapse imaging of neuroblast migration in acute slices of the adult mouse forebrain.

    PubMed

    Khlghatyan, Jivan; Saghatelyan, Armen

    2012-09-12

    There is a substantial body of evidence indicating that new functional neurons are constitutively generated from an endogenous pool of neural stem cells in restricted areas of the adult mammalian brain. Newborn neuroblasts from the subventricular zone (SVZ) migrate along the rostral migratory stream (RMS) to their final destination in the olfactory bulb (OB). In the RMS, neuroblasts migrate tangentially in chains ensheathed by astrocytic processes using blood vessels as a structural support and a source of molecular factors required for migration. In the OB, neuroblasts detach from the chains and migrate radially into the different bulbar layers where they differentiate into interneurons and integrate into the existing network. In this manuscript we describe the procedure for monitoring cell migration in acute slices of the rodent brain. The use of acute slices allows the assessment of cell migration in the microenvironment that closely resembling to in vivo conditions and in brain regions that are difficult to access for in vivo imaging. In addition, it avoids long culturing condition as in the case of organotypic and cell cultures that may eventually alter the migration properties of the cells. Neuronal precursors in acute slices can be visualized using DIC optics or fluorescent proteins. Viral labeling of neuronal precursors in the SVZ, grafting neuroblasts from reporter mice into the SVZ of wild-type mice, and using transgenic mice that express fluorescent protein in neuroblasts are all suitable methods for visualizing neuroblasts and following their migration. The later method, however, does not allow individual cells to be tracked for long periods of time because of the high density of labeled cells. We used a wide-field fluorescent upright microscope equipped with a CCD camera to achieve a relatively rapid acquisition interval (one image every 15 or 30 sec) to reliably identify the stationary and migratory phases. A precise identification of the duration of the stationary and migratory phases is crucial for the unambiguous interpretation of results. We also performed multiple z-step acquisitions to monitor neuroblasts migration in 3D. Wide-field fluorescent imaging has been used extensively to visualize neuronal migration. Here, we describe detailed protocol for labeling neuroblasts, performing real-time video-imaging of neuroblast migration in acute slices of the adult mouse forebrain, and analyzing cell migration. While the described protocol exemplified the migration of neuroblasts in the adult RMS, it can also be used to follow cell migration in embryonic and early postnatal brains.

  7. Silicon chip with capacitors and transistors for interfacing organotypic brain slice of rat hippocampus.

    PubMed

    Hutzler, Michael; Fromherz, Peter

    2004-04-01

    Probing projections between brain areas and their modulation by synaptic potentiation requires dense arrays of contacts for noninvasive electrical stimulation and recording. Semiconductor technology is able to provide planar arrays with high spatial resolution to be used with planar neuronal structures such as organotypic brain slices. To address basic methodical issues we developed a silicon chip with simple arrays of insulated capacitors and field-effect transistors for stimulation of neuronal activity and recording of evoked field potentials. Brain slices from rat hippocampus were cultured on that substrate. We achieved local stimulation of the CA3 region by applying defined voltage pulses to the chip capacitors. Recording of resulting local field potentials in the CA1 region was accomplished with transistors. The relationship between stimulation and recording was rationalized by a sheet conductor model. By combining a row of capacitors with a row of transistors we determined a simple stimulus-response matrix from CA3 to CA1. Possible contributions of inhomogeneities of synaptic projection, of tissue structure and of neuroelectronic interfacing were considered. The study provides the basis for a development of semiconductor chips with high spatial resolution that are required for long-term studies of topographic mapping.

  8. Developmental Social Cognitive Neuroscience: Insights from Deafness

    ERIC Educational Resources Information Center

    Corina, David; Singleton, Jenny

    2009-01-01

    The condition of deafness presents a developmental context that provides insight into the biological, cultural, and linguistic factors underlying the development of neural systems that impact social cognition. Studies of visual attention, behavioral regulation, language development, and face and human action perception are discussed. Visually…

  9. Moral Judgment Development across Cultures: Revisiting Kohlberg's Universality Claims

    ERIC Educational Resources Information Center

    Gibbs, John C.; Basinger, Karen S.; Grime, Rebecca L.; Snarey, John R.

    2007-01-01

    This article revisits Kohlberg's cognitive developmental claims that stages of moral judgment, facilitative processes of social perspective-taking, and moral values are commonly identifiable across cultures. Snarey [Snarey, J. (1985). "The cross-cultural universality of social-moral development: A critical review of Kohlbergian research."…

  10. Parents' Goals for Children: The Dynamic Coexistence of Individualism and Collectivism in Cultures and Individuals

    ERIC Educational Resources Information Center

    Tamis-LeMonda, Catherine S.; Way, Niobe; Hughes, Diane; Yoshikawa, Hirokazu; Kalman, Ronit Kahana; Niwa, Erika Y.

    2008-01-01

    Current scholarship on the cultural value systems of individualism and collectivism, and the associated developmental goals of autonomy and relatedness, has moved beyond grand divide theories to emphasize variation within individuals and cultures. We present a theoretical model on the dynamic coexistence of cultural value systems (at the macro…

  11. Perceived Parental Psychological Control and Adolescent Depressive Experiences: A Cross-Cultural Study with Belgian and South-Korean Adolescents

    ERIC Educational Resources Information Center

    Soenens, Bart; Park, Seong-Yeon; Vansteenkiste, Maarten; Mouratidis, Athanasios

    2012-01-01

    In recent research on psychologically controlling parenting, debate has arisen about the cross-cultural relevance of this construct, with some scholars arguing that the developmental outcomes of psychological control are culture-bound and others arguing that the detrimental effects of psychological control generalize across cultures. This study…

  12. Third Culture Kid Identification and Transitional Programs, Best Practices for School Administrators and Educators

    ERIC Educational Resources Information Center

    Simpson, Danielle S.

    2017-01-01

    This dissertation encapsulates a qualitative study that explores the identification and educational experiences of Third Culture Kids living abroad. A Third Culture Kid (TCK) is an individual who has spent a significant part of their developmental years outside of their parents' culture and country of nationality. This study specifically examines…

  13. Neighborhood and school ethnic structuring and cultural adaptations among Mexican-origin adolescents.

    PubMed

    White, Rebecca M B; Updegraff, Kimberly A; Umaña-Taylor, Adriana J; Zeiders, Katharine H; Perez-Brena, Norma; Burleson, Elizabeth

    2017-03-01

    The ethnic and racial structuring of U.S. neighborhoods may have important implications for developmental competencies during adolescence, including the development of heritage and mainstream cultural orientations. In particular, living in highly concentrated Latino neighborhoods during early adolescence-which channels adolescents into related school environments-may promote retention of the ethnic or heritage culture, but it also may constrain adaptation to the mainstream U.S. culture. We tested these hypotheses longitudinally in a sample of 246 Mexican origin adolescents (50.8% girls) and their parents. Data were collected 4 times over 8 years, with adolescents averaging 12.5 (SD = .58) to 19.6 (SD = .66) years of age across the period of the study. Latino ethnic concentration in early adolescents' neighborhoods promoted the retention of Mexican cultural orientations; Latino ethnic concentration in middle schools undermined the development of mainstream U.S cultural orientations. Findings are discussed in terms of integrating cultural-developmental theory with mainstream neighborhood theory to improve understandings of neighborhood and school ethnic concentration effects on adolescent development. (PsycINFO Database Record (c) 2017 APA, all rights reserved).

  14. Neighborhood and School Ethnic Structuring and Cultural Adaptations Among Mexican-Origin Adolescents

    PubMed Central

    White, Rebecca M. B.; Updegraff, Kimberly A.; Umaña-Taylor, Adriana J.; Zeiders, Katharine H.; Perez-Brena, Norma; Burleson, Elizabeth

    2016-01-01

    The ethnic and racial structuring of U.S. neighborhoods may have important implications for developmental competencies during adolescence, including the development of heritage and mainstream cultural orientations. In particular, living in highly concentrated Latino neighborhoods during early adolescence – which channels adolescents into related school environments – may promote retention of the ethnic or heritage culture, but it also may constrain adaptation to the mainstream U.S. culture. We tested these hypotheses longitudinally in a sample of 246 Mexican origin adolescents (50.8% girls) and their parents. Data were collected four times over eight years, with adolescents averaging 12.5 (SD = .58) to 19.6 (SD = .66) years of age across the period of the study. Latino ethnic concentration in early adolescents' neighborhoods promoted the retention of Mexican cultural orientations; Latino ethnic concentration in middle schools undermined the development of mainstream U.S cultural orientations. Findings are discussed in terms of integrating cultural-developmental theory with mainstream neighborhood theory to improve understandings of neighborhood and school ethnic concentration effects on adolescent development. PMID:27936822

  15. Difference or disorder? Cultural issues in understanding neurodevelopmental disorders.

    PubMed

    Norbury, Courtenay Frazier; Sparks, Alison

    2013-01-01

    Developmental disorders, such as autism spectrum disorder and specific language impairment, are biologically based disorders that currently rely on behaviorally defined criteria for diagnosis and treatment. Specific behaviors that are included in diagnostic frameworks and the point at which individual differences in behavior constitute abnormality are largely arbitrary decisions. Such decisions are therefore likely to be strongly influenced by cultural values and expectations. This is evident in the dramatically different prevalence rates of autism spectrum disorder across countries and across different ethnic groups within the same country. In this article, we critically evaluate the understanding of developmental disorders from a cultural perspective. We specifically consider the challenges of applying diagnostic methods across cultural contexts, the influence of cultural values and expectations on the identification and treatment of children with suspected disorders, and how cross-cultural studies can help to refine cognitive theories of disorder that have been derived exclusively from Western North American and European investigations. Our review synthesizes clinical, cultural, and theoretical work in this area, highlighting potential universals of disorder and concluding with recommendations for future research and practice.

  16. Ballistic delivery of dyes for structural and functional studies of the nervous system

    PubMed Central

    Gan, Wen-Biao; Grutzendler, Jaime; Wong, Rachel O.; Lichtman, Jeff W.

    2010-01-01

    This chapter describes a detail protocol for rapid labeling of cells in a variety of preparations by means of particle-mediated ballistic (gene gun) delivery of fluorescent dyes. This method has been used for rapid labeling of cells with either lipid or water-soluble dyes in a variety of preparations. In particular, carbocyanine lipophilic dyes such as DiI have been used to obtain Golgi-like labeling of neurons and glia in fixed and live cell cultures, brain slices, as well as fixed post-mortem human brain. Water-soluble calcium indicators such as calcium green-1 dextran have been used to image calcium dynamics in living brain slices and retinal explants. This ballistic labeling technique is thus useful for studying the structure and function of neurons and glia in both living and fixed specimens. PMID:20147144

  17. Developmental Dyscalculia and Low Numeracy in Chinese Children

    ERIC Educational Resources Information Center

    Chan, Winnie Wai Lan; Au, Terry K.; Tang, Joey

    2013-01-01

    Children struggle with mathematics for different reasons. Developmental dyscalculia and low numeracy--two kinds of mathematical difficulties--may have their roots, respectively, in poor understanding of exact non-symbolic numerosities and of symbolic numerals. This study was the first to explore whether Chinese children, despite cultural and…

  18. Integrating Academic Information into Developmental Writing Courses.

    ERIC Educational Resources Information Center

    Troiano, Edna M.; Draus, Julia

    In 1983, Charles County Community College (CCCC) initiated a project to infuse academic information from the sciences, humanities, and social sciences into developmental courses. The reorganization assignments related to 27 topics that promoted academic examination and cultural literacy while at the same time drawing from students' own…

  19. Linking Social Change and Developmental Change: Shifting Pathways of Human Development

    ERIC Educational Resources Information Center

    Greenfield, Patricia M.

    2009-01-01

    P. M. Greenfield's new theory of social change and human development aims to show how changing sociodemographic ecologies alter cultural values and learning environments and thereby shift developmental pathways. Worldwide sociodemographic trends include movement from rural residence, informal education at home, subsistence economy, and…

  20. Navigating Cultural Worlds and Negotiating Identities: A Conceptual Model

    ERIC Educational Resources Information Center

    Mistry, Jayanthi; Wu, Jean

    2010-01-01

    For children from culturally and linguistically diverse backgrounds the ability to maintain flexible identities and integrate multiple facets of self is a crucial developmental task. We present a conceptual model for the development of expertise in navigating across cultures, delineating how community characteristics interact with family and…

  1. Use and perception of the environment: cultural and developmental processes

    Treesearch

    Martin M. Chemers; Irwin Altman

    1977-01-01

    This paper presents a "social systems" orientation for integrating the diverse aspects of environment, culture, and individual behavior. It suggests that a wide range of variables, including the physical environment, cultural and social processes, environmental perceptions and cognitions, behavior, and products of behavior, are connected in a complex,...

  2. Cultural Adaptation of a Preventive Program for Ultra-Orthodox Preschool Boys

    ERIC Educational Resources Information Center

    Gilboa, Yafit

    2016-01-01

    Cultural factors significantly influence the effectiveness of pediatric screening that enables the prevention of developmental disturbances. The formulation of intervention programs must match the needs of the child, his or her family, and educators. Recognizing the importance of creating an intervention program accessible to the culture of the…

  3. Patterns of Cultural Adjustment among Young Migrants to Australia

    ERIC Educational Resources Information Center

    Sonderegger, Robi; Barrett, Paula M.

    2004-01-01

    In response to appeals for empirical data on culture-specific differences and developmental pathways of acculturative stress among young migrants and refugees, the present study examines the cultural adjustment patterns of ethnically diverse migrants to Australia. Two hundred and seventy three primary and high school students (comprised of…

  4. The rat whole embryo culture assay using the Dysmorphology Score system.

    PubMed

    Zhang, Cindy; Panzica-Kelly, Julie; Augustine-Rauch, Karen

    2013-01-01

    The rat whole embryo culture (WEC) system has been used extensively for characterizing teratogenic properties of test chemicals. In this chapter, we describe the methodology for culturing rat embryos as well as a new morphological score system, the Dysmorphology Score (DMS) system for assessing morphology of mid gestation (gestational day 11) rat embryos. In contrast to the developmental stage focused scoring associated with the Brown and Fabro score system, this new score system assesses the respective degree of severity of dysmorphology, which delineates normal from abnormal morphology of specific embryonic structures and organ systems. This score system generates an approach that allows rapid identification and quantification of adverse developmental findings, making it conducive for characterization of compounds for teratogenic properties and screening activities.

  5. Anti-Yo antibody uptake and interaction with its intracellular target antigen causes Purkinje cell death in rat cerebellar slice cultures: a possible mechanism for paraneoplastic cerebellar degeneration in humans with gynecological or breast cancers.

    PubMed

    Greenlee, John E; Clawson, Susan A; Hill, Kenneth E; Wood, Blair; Clardy, Stacey L; Tsunoda, Ikuo; Carlson, Noel G

    2015-01-01

    Anti-Yo antibodies are immunoglobulin G (IgG) autoantibodies reactive with a 62 kDa Purkinje cell cytoplasmic protein. These antibodies are closely associated with paraneoplastic cerebellar degeneration in the setting of gynecological and breast malignancies. We have previously demonstrated that incubation of rat cerebellar slice cultures with patient sera and cerebrospinal fluid containing anti-Yo antibodies resulted in Purkinje cell death. The present study addressed three fundamental questions regarding the role of anti-Yo antibodies in disease pathogenesis: 1) Whether the Purkinje cell cytotoxicity required binding of anti-Yo antibody to its intraneuronal 62 kDa target antigen; 2) whether Purkinje cell death might be initiated by antibody-dependent cellular cytotoxicity rather than intracellular antibody binding; and 3) whether Purkinje cell death might simply be a more general result of intracellular antibody accumulation, rather than of specific antibody-antigen interaction. In our study, incubation of rat cerebellar slice cultures with anti-Yo IgG resulted in intracellular antibody binding, and cell death. Infiltration of the Purkinje cell layer by cells of macrophage/microglia lineage was not observed until extensive cell death was already present. Adsorption of anti-Yo IgG with its 62 kDa target antigen abolished both antibody accumulation and cytotoxicity. Antibodies to other intracellular Purkinje cell proteins were also taken up by Purkinje cells and accumulated intracellularly; these included calbindin, calmodulin, PCP-2, and patient anti-Purkinje cell antibodies not reactive with the 62 kDa Yo antigen. However, intracellular accumulation of these antibodies did not affect Purkinje cell viability. The present study is the first to demonstrate that anti-Yo antibodies cause Purkinje cell death by binding to the intracellular 62 kDa Yo antigen. Anti-Yo antibody cytotoxicity did not involve other antibodies or factors present in patient serum and was not initiated by brain mononuclear cells. Purkinje cell death was not simply due to intraneuronal antibody accumulation.

  6. Anti-Yo Antibody Uptake and Interaction with Its Intracellular Target Antigen Causes Purkinje Cell Death in Rat Cerebellar Slice Cultures: A Possible Mechanism for Paraneoplastic Cerebellar Degeneration in Humans with Gynecological or Breast Cancers

    PubMed Central

    Greenlee, John E.; Clawson, Susan A.; Hill, Kenneth E.; Wood, Blair; Clardy, Stacey L.; Tsunoda, Ikuo; Carlson, Noel G.

    2015-01-01

    Anti-Yo antibodies are immunoglobulin G (IgG) autoantibodies reactive with a 62 kDa Purkinje cell cytoplasmic protein. These antibodies are closely associated with paraneoplastic cerebellar degeneration in the setting of gynecological and breast malignancies. We have previously demonstrated that incubation of rat cerebellar slice cultures with patient sera and cerebrospinal fluid containing anti-Yo antibodies resulted in Purkinje cell death. The present study addressed three fundamental questions regarding the role of anti-Yo antibodies in disease pathogenesis: 1) Whether the Purkinje cell cytotoxicity required binding of anti-Yo antibody to its intraneuronal 62 kDa target antigen; 2) whether Purkinje cell death might be initiated by antibody-dependent cellular cytotoxicity rather than intracellular antibody binding; and 3) whether Purkinje cell death might simply be a more general result of intracellular antibody accumulation, rather than of specific antibody-antigen interaction. In our study, incubation of rat cerebellar slice cultures with anti-Yo IgG resulted in intracellular antibody binding, and cell death. Infiltration of the Purkinje cell layer by cells of macrophage/microglia lineage was not observed until extensive cell death was already present. Adsorption of anti-Yo IgG with its 62 kDa target antigen abolished both antibody accumulation and cytotoxicity. Antibodies to other intracellular Purkinje cell proteins were also taken up by Purkinje cells and accumulated intracellularly; these included calbindin, calmodulin, PCP-2, and patient anti-Purkinje cell antibodies not reactive with the 62 kDa Yo antigen. However, intracellular accumulation of these antibodies did not affect Purkinje cell viability. The present study is the first to demonstrate that anti-Yo antibodies cause Purkinje cell death by binding to the intracellular 62 kDa Yo antigen. Anti-Yo antibody cytotoxicity did not involve other antibodies or factors present in patient serum and was not initiated by brain mononuclear cells. Purkinje cell death was not simply due to intraneuronal antibody accumulation. PMID:25885452

  7. Activation of Intrinsic Immune Responses and Microglial Phagocytosis in an Ex Vivo Spinal Cord Slice Culture Model of West Nile Virus Infection

    PubMed Central

    Quick, Eamon D.; Leser, J. Smith; Tyler, Kenneth L.

    2014-01-01

    ABSTRACT West Nile virus (WNV) is a neurotropic flavivirus that causes significant neuroinvasive disease involving the brain and/or spinal cord. Experimental mouse models of WNV infection have established the importance of innate and adaptive immune responses in controlling the extent and severity of central nervous system (CNS) disease. However, differentiating between immune responses that are intrinsic to the CNS and those that are dependent on infiltrating inflammatory cells has proven difficult. We used a murine ex vivo spinal cord slice culture (SCSC) model to determine the innate immune processes specific to the CNS during WNV infections. By 7 days after ex vivo infection of SCSCs, the majority of neurons and a substantial percentage of astrocytes were infected with WNV, resulting in apoptotic cell death and astrogliosis. Microglia, the resident immune cells of the CNS, were activated by WNV infection, as exemplified by their amoeboid morphology, the development of filopodia and lamellipodia, and phagocytosis of WNV-infected cells and debris. Microglial cell activation was concomitant with increased expression of proinflammatory cytokines and chemokines, including CXCL10, CXCL1, CCL5, CCL3, CCL2, tumor necrosis factor alpha (TNF-α), TNF-related apoptosis-inducing ligand (TRAIL), and interleukin-6 (IL-6). The application of minocycline, an inhibitor of neuroinflammation, altered the WNV-induced proinflammatory cytokine/chemokine expression profile, with inhibited production of CCL5, CCL2, and IL-6. Our findings establish that CNS-resident cells have the capacity to initiate a robust innate immune response against WNV infection in the absence of infiltrating inflammatory cells and systemic immune responses. IMPORTANCE There are no specific treatments of proven efficacy available for WNV neuroinvasive disease. A better understanding of the pathogenesis of WNV CNS infection is crucial for the rational development of novel therapies. Development of a spinal cord slice culture (SCSC) model facilitates the study of WNV pathogenesis and allows investigation of the intrinsic immune responses of the CNS. Our studies demonstrate that robust CNS innate immune responses, including microglial activation and proinflammatory cytokine/chemokine production, develop independently of contributions from the peripheral immune system and CNS-infiltrating inflammatory cells. PMID:25165111

  8. A View of Developmental Education and the "Disadvantaged" in a National Perspective: Implications for International Education

    ERIC Educational Resources Information Center

    Wilson, Thomasyne Lightfoote

    1972-01-01

    After defining developmental education as referring to processes that expand the physical, emotional, educational, artistic, scientific, and political spheres of diverse ethnic and cultural communities within a nation, the author discusses the case of the Loma people of Liberia. (JM)

  9. The Developmental Model of Intercultural Sensitivity: A Tool for Understanding Principals' Cultural Competence

    ERIC Educational Resources Information Center

    Hernandez, Frank; Kose, Brad W.

    2012-01-01

    Principals' understanding and skills pertaining to diversity are important in leading diverse schools and preparing all students for a democratic and multicultural society. Although educational leadership scholars have theorized about exemplary leadership of and for diversity, a developmental perspective on principals' diversity or cultural…

  10. Supporting Children and Adolescents with Deployed Caregivers: A Structured Group Approach for School Counselors

    ERIC Educational Resources Information Center

    Rush, Christina Mitchell; Akos, Patrick

    2007-01-01

    Children and adolescents of deployed soldiers require preventive and sometimes remedial service from school counselors, because deployment taxes coping skills and exacerbates normal developmental challenges. This article reviews the cultural and developmental context for children with deployed caregivers and outlines a ten session, hybrid…

  11. Diversity & Developmentally Appropriate Practices: Challenges for Early Childhood Education.

    ERIC Educational Resources Information Center

    Mallory, Bruce L., Ed.; New, Rebecca S., Ed.

    The current conceptualization of what is appropriate in early childhood education is overly narrow in its interpretation of the role of the teacher, and with respect to variations in cultural and developmental diversity. Based on this observation, this book discusses various issues surrounding diversity, inclusion, and appropriate early…

  12. Perception of Individualism and Collectivism in Dutch Society: A Developmental Approach

    ERIC Educational Resources Information Center

    Oppenheimer, Louis

    2004-01-01

    Within Triandis's (1994) theoretical framework, two studies are reported that deal with the developmental course for subjective perceptions of cultural dimensions in Dutch society (i.e., vertical and horizontal individualism and collectivism). While perceptions of society are always subjectively determined, the perceived dimensions that are…

  13. Treatment Strategies for Hispanic Developmentally Disabled Clients.

    ERIC Educational Resources Information Center

    Omdahl, Doris

    Professionals who work with developmentally disabled Hispanic clients must enhance their own ability to work with ethnic minorities and become thoroughly familiar with the culture of the people they serve. To encourage disabled Hispanic individuals to apply for services, agencies can employ strategies such as bilingual capability, use of…

  14. Schools as Developmental Contexts during Adolescence

    ERIC Educational Resources Information Center

    Eccles, Jacquelynne S.; Roeser, Robert W.

    2011-01-01

    Considerable strides have been made in the past decade in recognizing the centrality of the cultural context of schooling to adolescent development. In this review, adopting a developmental systems conceptualization of schooling, we focus on selected new research findings from the past decade regarding how (a) teachers, curricular tasks, and…

  15. Cultural and Cognitive Development in Short-Term Study Abroad: Illuminating the 360 Experience

    ERIC Educational Resources Information Center

    Burton, Susan

    2012-01-01

    The three articles in this dissertation investigate leading others through developmental opportunities by facilitating their engagement in intercultural challenges. Specifically the research explores the meaning followers make of developmental challenges during short-term study abroad experiences and encounters with diversity. Data in the form of…

  16. Getting Back to the Woods: Familial Perspectives on Culture and Preschoolers' Acquisition of Self-Regulation and Emotion Regulation

    ERIC Educational Resources Information Center

    Boyer, Wanda

    2013-01-01

    Discourse on culture is vital to early childhood educators' understanding of the young child in various socio-cultural experiences in family and community settings. In this article, the author will present a contemporary definition of culture. This article will then discuss the developmental constructs of self-regulation and emotion regulation and…

  17. Temporal variation and interaction between nutritional and developmental instability in prehistoric Japanese populations.

    PubMed

    Hoover, Kara C; Matsumura, Hirofumi

    2008-12-01

    We examined nutritional and developmental instability in prehistoric Japan, using data from 49 individuals across 13 archaeological sites. Hypoplasia incidence was used as a measure of nutritional stress, and fluctuating asymmetry (of upper facial breath, orbital breadth, and orbital height) as an indirect assessment of developmental instability. Abundant resources due to a stable climate during the Middle Jomon (5,000-3,000 BP) encouraged population growth, which led to regional cultural homogeneity and complexity. A population crash on Honshu in the Late/Final Jomon (roughly 4,000-2,000 BP) led to regionally divergent subsistence economies and settlement patterns. We find that the nutritional stress was consistent between periods, but developmental instability (DI) decreased in the Late/Final Jomon. While the DI values were not statistically significant, the higher values for Middle Jomon may result from sedentism, social stratification, and differential access to resources. On Hokkaido, Jomon culture persisted until the Okhotsk period (1,000-600 BP), marked by the arrival of immigrants from Sakhalin. Nutritional stress was consistent between Middle and Late/Final Jomon, but DI increased in the Late/Final. Nutritional and developmental instability decreased from Late/Final to Okhotsk, suggesting a positive immigrant effect. We expected to find an association between stress markers due to the synergistic relationship between nutrition and pathology. The data support this hypothesis, but only one finding was statistically significant. While high critical values from small sample sizes place limits on the significance of our results, we find that the impact of environmental and cultural change to prehistoric Japanese populations was minimal. (c) 2008 Wiley-Liss, Inc.

  18. Enuresis: An Historical, Cultural, and Contemporary Account of Etiology and Treatment

    ERIC Educational Resources Information Center

    McDonald, James E.; Trepper, Terry

    1977-01-01

    The authors discuss the cultural relativism of enuresis and the subsequent notion that urinary incontinence may not be a disease or psychopathy, but, rather, a problem associated with social expectations and developmental delays. (Author)

  19. Review: role of carbon sources for in vitro plant growth and development.

    PubMed

    Yaseen, Mehwish; Ahmad, Touqeer; Sablok, Gaurav; Standardi, Alvaro; Hafiz, Ishfaq Ahmad

    2013-04-01

    In vitro plant cells, tissues and organ cultures are not fully autotrophic establishing a need for carbohydrates in culture media to maintain the osmotic potential, as well as to serve as energy and carbon sources for developmental processes including shoot proliferation, root induction as well as emission, embryogenesis and organogenesis, which are highly energy demanding developmental processes in plant biology. A variety of carbon sources (both reducing and non-reducing) are used in culture media depending upon genotypes and specific stages of growth. However, sucrose is most widely used as a major transport-sugar in the phloem sap of many plants. In micropropagation systems, morphogenetic potential of plant tissues can greatly be manipulated by varying type and concentration of carbon sources. The present article reviews the past and current findings on carbon sources and their sustainable utilization for in vitro plant tissue culture to achieve better growth rate and development.

  20. Cultural Influences on Perceptions of Health, Illness, and Disability: A Review and Focus on Autism

    ERIC Educational Resources Information Center

    Ravindran, Neeraja; Myers, Barbara J.

    2012-01-01

    This conceptual paper considers the role of culture in shaping family, professional, and community understanding of developmental disabilities and their treatments. The meanings of health, illness, and disability vary greatly across cultures and across time. We use Bronfenbrenner's ecological model to provide a theoretical framework for examining…

  1. Cross-Cultural Psychology's Challenges to Our Ideas of Children and Development.

    ERIC Educational Resources Information Center

    American Psychologist, 1979

    1979-01-01

    The Laboratory of Comparative Human Cognition has found that, if cross-cultural psychology has mounted a challenge to developmental psychology, it is because it has forced recognition of the fact that no context of observation, including the laboratory, is culturally neutral. Settings for behavior are embedded in larger systems of social…

  2. Traditional Media and Indigenous Culture: Rethinking Developmental Relationships.

    ERIC Educational Resources Information Center

    Roe, Robert C.

    Over 60 years of American anthropological and sociological research writings have erected a concept of culture that restricts its role in planned change models to one of active or tacit resistance to change. As an alternative, this paper suggests that the substitution of a contemporary, dynamic conceptualization of culture as developed by…

  3. The Cultural Context of Infancy. Volume 1: Biology, Culture, and Infant Development.

    ERIC Educational Resources Information Center

    Nugent, J. Kevin, Ed.; And Others

    Noting that patterns of childrearing are highly variable across the human species, this book explores the developmental processes of infancy over a wide range of cultural and social environments. By presenting multiple alternative examples of the context of infant development, the book attempts to stimulate continued discussion on the ways in…

  4. Parenting Beliefs, Behaviors, and Parent-Child Relations: A Cross-Cultural Perspective

    ERIC Educational Resources Information Center

    Rubin, Kenneth H., Ed.; Chung, Ock Boon, Ed.

    2006-01-01

    The purpose of this book, is to present a rather simple argument. Parents' thoughts about childrearing and the ways in which they interact with children to achieve particular parenting or developmental goals, are culturally determined. Within any culture, children are shaped by the physical and social settings within which they live, culturally…

  5. Effect of Culture Conditions on Viability of Mouse and Rat Embryos Developed in Vitro

    PubMed Central

    Popova, Elena; Bader, Michael; Krivokharchenko, Alexander

    2011-01-01

    Currently in vitro culture of mouse preimplantation embryos has become a very important technique to investigate different mechanisms of early embryogenesis. However, there is a big difference in the preimplantation development between mammalian species. Despite close relatedness to mice, in vitro cultivation of rat preimplantation embryos is still delicate and needs further investigation and optimizations. In this study we have compared the in vitro developmental potential of mouse and rat embryos cultured at different culture conditions in parallel experiments. Interestingly, mouse zygotes developed in vitro until blastocyst stage even in inadequate medium without any phosphates and with low osmolarity which was formulated especially for cultivation of rat embryos. Rat parthenotes and zygotes developed in M16 medium formulated for mouse embryos only till 2-cell stage and further development is blocked completely at this stage. Moreover, developmental ability of rat embryos in vitro was significantly lower in comparison with mouse even in special rat mR1ECM medium. Mouse and rat embryos at 2-cell stage obtained in vivo developed until blastocyst stages significantly more efficiently compared to zygotes. Culture of mouse zygotes in glass capillaries resulted in a significantly higher rate of morula and blastocyst development compared with dishes. The Well-of-the-Well system resulted in a significant improvement when compared with dishes for the culture of rat zygotes only until morula stage. Reduced oxygen tension increased the developmental rate of rat but not mouse zygotes until blastocyst stage. This study demonstrates that development of early preimplantation embryos is altered by different culture conditions and show strong differences even between two related species such as mice and rats. Therefore, for understanding the fundamental mechanisms of early mammalian development it is very important to use embryos of various species. PMID:24710194

  6. The Formative Method for Adapting Psychotherapy (FMAP): A community-based developmental approach to culturally adapting therapy

    PubMed Central

    Hwang, Wei-Chin

    2010-01-01

    How do we culturally adapt psychotherapy for ethnic minorities? Although there has been growing interest in doing so, few therapy adaptation frameworks have been developed. The majority of these frameworks take a top-down theoretical approach to adapting psychotherapy. The purpose of this paper is to introduce a community-based developmental approach to modifying psychotherapy for ethnic minorities. The Formative Method for Adapting Psychotherapy (FMAP) is a bottom-up approach that involves collaborating with consumers to generate and support ideas for therapy adaptation. It involves 5-phases that target developing, testing, and reformulating therapy modifications. These phases include: (a) generating knowledge and collaborating with stakeholders (b) integrating generated information with theory and empirical and clinical knowledge, (c) reviewing the initial culturally adapted clinical intervention with stakeholders and revising the culturally adapted intervention, (d) testing the culturally adapted intervention, and (e) finalizing the culturally adapted intervention. Application of the FMAP is illustrated using examples from a study adapting psychotherapy for Chinese Americans, but can also be readily applied to modify therapy for other ethnic groups. PMID:20625458

  7. Prolonged Mek1/2 suppression impairs the developmental potential of embryonic stem cells.

    PubMed

    Choi, Jiho; Huebner, Aaron J; Clement, Kendell; Walsh, Ryan M; Savol, Andrej; Lin, Kaixuan; Gu, Hongcang; Di Stefano, Bruno; Brumbaugh, Justin; Kim, Sang-Yong; Sharif, Jafar; Rose, Christopher M; Mohammad, Arman; Odajima, Junko; Charron, Jean; Shioda, Toshi; Gnirke, Andreas; Gygi, Steven; Koseki, Haruhiko; Sadreyev, Ruslan I; Xiao, Andrew; Meissner, Alexander; Hochedlinger, Konrad

    2017-08-10

    Concomitant activation of the Wnt pathway and suppression of Mapk signalling by two small molecule inhibitors (2i) in the presence of leukaemia inhibitory factor (LIF) (hereafter termed 2i/L) induces a naive state in mouse embryonic stem (ES) cells that resembles the inner cell mass (ICM) of the pre-implantation embryo. Since the ICM exists only transiently in vivo, it remains unclear how sustained propagation of naive ES cells in vitro affects their stability and functionality. Here we show that prolonged culture of male mouse ES cells in 2i/L results in irreversible epigenetic and genomic changes that impair their developmental potential. Furthermore, we find that female ES cells cultured in conventional serum plus LIF medium phenocopy male ES cells cultured in 2i/L. Mechanistically, we demonstrate that the inhibition of Mek1/2 is predominantly responsible for these effects, in part through the downregulation of DNA methyltransferases and their cofactors. Finally, we show that replacement of the Mek1/2 inhibitor with a Src inhibitor preserves the epigenetic and genomic integrity as well as the developmental potential of ES cells. Taken together, our data suggest that, although short-term suppression of Mek1/2 in ES cells helps to maintain an ICM-like epigenetic state, prolonged suppression results in irreversible changes that compromise their developmental potential.

  8. Glucose Regulates Hypothalamic Long-chain Fatty Acid Metabolism via AMP-activated Kinase (AMPK) in Neurons and Astrocytes*

    PubMed Central

    Taïb, Bouchra; Bouyakdan, Khalil; Hryhorczuk, Cécile; Rodaros, Demetra; Fulton, Stephanie; Alquier, Thierry

    2013-01-01

    Hypothalamic controls of energy balance rely on the detection of circulating nutrients such as glucose and long-chain fatty acids (LCFA) by the mediobasal hypothalamus (MBH). LCFA metabolism in the MBH plays a key role in the control of food intake and glucose homeostasis, yet it is not known if glucose regulates LCFA oxidation and esterification in the MBH and, if so, which hypothalamic cell type(s) and intracellular signaling mechanisms are involved. The aim of this study was to determine the impact of glucose on LCFA metabolism, assess the role of AMP-activated Kinase (AMPK), and to establish if changes in LCFA metabolism and its regulation by glucose vary as a function of the kind of LCFA, cell type, and brain region. We show that glucose inhibits palmitate oxidation via AMPK in hypothalamic neuronal cell lines, primary hypothalamic astrocyte cultures, and MBH slices ex vivo but not in cortical astrocytes and slice preparations. In contrast, oleate oxidation was not affected by glucose or AMPK inhibition in MBH slices. In addition, our results show that glucose increases palmitate, but not oleate, esterification into neutral lipids in neurons and MBH slices but not in hypothalamic astrocytes. These findings reveal for the first time the metabolic fate of different LCFA in the MBH, demonstrate AMPK-dependent glucose regulation of LCFA oxidation in both astrocytes and neurons, and establish metabolic coupling of glucose and LCFA as a distinguishing feature of hypothalamic nuclei critical for the control of energy balance. PMID:24240094

  9. AN ORGANOTYPIC UNIAXIAL STRAIN MODEL USING MICROFLUIDICS

    PubMed Central

    Dollé, Jean-Pierre; Morrison, Barclay; Schloss, Rene R.; Yarmush, Martin L.

    2012-01-01

    Traumatic brain injuries are the leading cause of disability each year in the US. The most common and devastating consequence is the stretching of axons caused by shear deformation that occurs during rotational acceleration of the brain during injury. The injury effects on axonal molecular and functional events are not fully characterized. We have developed a strain injury model that maintains the three dimensional cell architecture and neuronal networks found in vivo with the ability to visualize individual axons and their response to a mechanical injury. The advantage of this model is that it can apply uniaxial strains to axons that make functional connections between two organotypic slices and injury responses can be observed in real-time and over long term. This uniaxial strain model was designed to be capable of applying an array of mechanical strains at various rates of strain, thus replicating a range of modes of axonal injury. Long term culture, preservation of slice and cell orientation, and slice-slice connection on the device was demonstrated. The device has the ability to strain either individual axons or bundles of axons through the control of microchannel dimensions. The fidelity of the model was verified by observing characteristic responses to various strain injuries which included axonal beading, delayed elastic effects and breakdown in microtubules. Microtubule breakdown was shown to be dependent on the degree of the applied strain field, where maximal breakdown was observed at peak strain and minimal breakdown is observed at low strain. This strain injury model could be a powerful tool in assessing strain injury effects on functional axonal connections. PMID:23233120

  10. Redox regulation of epithelial sodium channels examined in alveolar type 1 and 2 cells patch-clamped in lung slice tissue.

    PubMed

    Helms, My N; Jain, Lucky; Self, Julie L; Eaton, Douglas C

    2008-08-15

    The alveolar surface of the lung is lined by alveolar type 1 (AT1) and type 2 (AT2) cells. Using single channel patch clamp analysis in lung slice preparations, we are able to uniquely study AT1 and AT2 cells separately from intact lung. We report for the first time the Na+ transport properties of type 2 cells accessed in live lung tissue (as we have done in type 1 cells). Type 2 cells in lung tissue slices express both highly selective cation and nonselective cation channels with average conductances of 8.8 +/- 3.2 and 22.5 +/- 6.3 picosiemens, respectively. Anion channels with 10-picosiemen conductance are also present in the apical membrane of type 2 cells. Our lung slice studies importantly verify the use of cultured cell model systems commonly used in lung epithelial sodium channel (ENaC) studies. Furthermore, we identify novel functional differences between the cells that make up the alveolar epithelium. One important difference is that exposure to the nitric oxide (NO) donor, PAPA-NONOate (1.5 microm), significantly decreases average ENaC NPo in type 2 cells (from 1.38 +/- 0.26 to 0.82 +/- 0.16; p < 0.05 and n = 18) but failed to alter ENaC activity in alveolar type 1 cells. Elevating endogenous superoxide (O2.) levels with Ethiolat, a superoxide dismutase inhibitor, prevented NO inhibition of ENaC activity in type 2 cells, supporting the novel hypothesis that O2. and NO signaling plays an important role in maintaining lung fluid balance.

  11. Optimized heterologous transfection of viable adult organotypic brain slices using an enhanced gene gun

    PubMed Central

    2013-01-01

    Background Organotypic brain slices (OTBS) are an excellent experimental compromise between the facility of working with cell cultures and the biological relevance of using animal models where anatomical, morphological, and cellular function of specific brain regions can be maintained. The biological characteristics of OTBS can subsequently be examined under well-defined conditions. They do, however, have a number of limitations; most brain slices are derived from neonatal animals, as it is difficult to properly prepare and maintain adult OTBS. There are ample problems with tissue integrity as OTBS are delicate and frequently become damaged during the preparative stages. Notwithstanding these obstacles, the introduced exogenous proteins into both neuronal cells, and cells imbedded within tissues, have been consistently difficult to achieve. Results Following the ex vivo extraction of adult mouse brains, mounted inside a medium-agarose matrix, we have exploited a precise slicing procedure using a custom built vibroslicer. To transfect these slices we used an improved biolistic transfection method using a custom made low-pressure barrel and novel DNA-coated nanoparticles (40 nm), which are drastically smaller than traditional microparticles. These nanoparticles also minimize tissue damage as seen by a significant reduction in lactate dehydrogenase activity as well as propidium iodide (PI) and dUTP labelling compared to larger traditional gold particles used on these OTBS. Furthermore, following EYFP exogene delivery by gene gun, the 40 nm treated OTBS displayed a significantly larger number of viable NeuN and EYFP positive cells. These OTBS expressed the exogenous proteins for many weeks. Conclusions Our described methodology of producing OTBS, which results in better reproducibility with less tissue damage, permits the exploitation of mature fully formed adult brains for advanced neurobiological studies. The novel 40 nm particles are ideal for the viable biolistic transfection of OTBS by reducing tissue stress while maintaining long term exogene expression. PMID:24354851

  12. Single-molecule RNA detection at depth by hybridization chain reaction and tissue hydrogel embedding and clearing.

    PubMed

    Shah, Sheel; Lubeck, Eric; Schwarzkopf, Maayan; He, Ting-Fang; Greenbaum, Alon; Sohn, Chang Ho; Lignell, Antti; Choi, Harry M T; Gradinaru, Viviana; Pierce, Niles A; Cai, Long

    2016-08-01

    Accurate and robust detection of mRNA molecules in thick tissue samples can reveal gene expression patterns in single cells within their native environment. Preserving spatial relationships while accessing the transcriptome of selected cells is a crucial feature for advancing many biological areas - from developmental biology to neuroscience. However, because of the high autofluorescence background of many tissue samples, it is difficult to detect single-molecule fluorescence in situ hybridization (smFISH) signals robustly in opaque thick samples. Here, we draw on principles from the emerging discipline of dynamic nucleic acid nanotechnology to develop a robust method for multi-color, multi-RNA imaging in deep tissues using single-molecule hybridization chain reaction (smHCR). Using this approach, single transcripts can be imaged using epifluorescence, confocal or selective plane illumination microscopy (SPIM) depending on the imaging depth required. We show that smHCR has high sensitivity in detecting mRNAs in cell culture and whole-mount zebrafish embryos, and that combined with SPIM and PACT (passive CLARITY technique) tissue hydrogel embedding and clearing, smHCR can detect single mRNAs deep within thick (0.5 mm) brain slices. By simultaneously achieving ∼20-fold signal amplification and diffraction-limited spatial resolution, smHCR offers a robust and versatile approach for detecting single mRNAs in situ, including in thick tissues where high background undermines the performance of unamplified smFISH. © 2016. Published by The Company of Biologists Ltd.

  13. Acceptance of Disability and Hwa-Byung among Korean Mothers of Children with Developmental Disabilities

    ERIC Educational Resources Information Center

    Kim, Eunha; Hwang, Jowon; Park, Sukyoung

    2018-01-01

    This study examined how child problem behaviour could be related to maternal Hwa-Byung (HB; Korean culture syndrome, meaning "anger illness") among Korean mothers of children with developmental disabilities. Acceptance of disabilities and parenting stress were tested as mediators for the relationship between child problem behaviour and…

  14. Cultural Responsivity in Clinical Psychology Graduate Students: A Developmental Approach to the Prediction of Learning

    ERIC Educational Resources Information Center

    Berrin, Sebastian Everett

    2010-01-01

    This study used a mixed-method approach to examine students' experiences in multicultural training and their opinions about various aspects of their course(s). A developmental model of learning was employed to analyze results. More specifically, this study explored the relationship between clinical psychology doctoral students' self-reported…

  15. Depression in China: Integrating Developmental Psychopathology and Cultural-Clinical Psychology

    ERIC Educational Resources Information Center

    Ryder, Andrew G.; Sun, Jiahong; Zhu, Xiongzhao; Yao, Shuqiao; Chentsova-Dutton, Yulia E.

    2012-01-01

    With a starting point in John Abela's groundbreaking developmental psychopathology research on adolescent depression in China, we aimed to review the state of the literature on Chinese depression across the lifespan. We began with Dr. Abela's published studies relevant to depression in China and our own research with adults before turning to the…

  16. Maternal Perceptions of Care-Giving of Children with Developmental Disabilities in the United Arab Emirates

    ERIC Educational Resources Information Center

    Crabtree, Sara Ashencaen

    2007-01-01

    Background: This qualitative study discusses family care-giving of children with developmental disabilities in the United Arab Emirates. Findings were reviewed in terms of the influence of culture on gender norms governing parental behaviour and aspirations. Method: An ethnographic approach is used in which parent participants were interviewed…

  17. Lifespan Development of Neuromodulation of Adaptive Control and Motivation as an Ontogenetic Mechanism for Developmental Niche Construction

    ERIC Educational Resources Information Center

    Li, Shu-Chen

    2013-01-01

    Instead of viewing organisms and individuals as passive recipients of their biological, ecological, and cultural inheritances, the developmental niche construction theory and the biocultural co-construction framework both emphasize that the individual's agency plays a key role in regulating how environmental and sociocontextual influences may…

  18. Socialization Ambiguity in Samoan Adolescents: A Model for Human Development and Stress in the Context of Culture Change

    ERIC Educational Resources Information Center

    McDade, Thomas W.; Worthman, Carol M.

    2004-01-01

    Rapid globalization is forcing youth around the world to confront new developmental challenges, and conceptual models are needed that can capture this experience and its developmental implications. Exposure to nontraditional lifestyles opens up new socialization opportunities and raises the possibility of stress-inducing dissonance between…

  19. Gender and Cultural Diversity Bias in Developmental Textbooks.

    ERIC Educational Resources Information Center

    Conti, Nancy E.; Kimmel, Ellen B.

    This paper reports the results of a content-analysis for the treatment of gender and diversity in the 11 top-selling lifespan developmental textbooks. The purposes were to measure the amount of information provided on aspects of development specific to Caucasian females and females of Color and to evaluate qualitatively the incorporation of the…

  20. Effects of cilostamide and/or forskolin on the meiotic resumption and development competence of growing ovine oocytes selected by brilliant cresyl blue staining.

    PubMed

    Azari-Dolatabad, Nima; Rahmani, H R; Hajian, M; Ostadhosseini, S; Hosseini, S M; Nasr-Esfahani, M H

    2016-05-01

    The relevance of low developmental competence of in vitro-matured oocyte to the incomplete/delayed cytoplasmic maturation, and the heterogeneity of retrieved oocytes is well established in several species. A short phase of prematuration culture was used to allow better oocyte cytoplasmic maturation. The preselection of growing and fully grown oocytes has been proposed to improve developmental competency. This study investigated the effects of phosphodiesterase type 3-specific inhibitor, cilostamide, and adenylate cyclase activator, forskolin, on the resumption of meiosis and developmental competence of growing ovine oocytes selected by brilliant cresyl blue (BCB) staining. Results indicate that cilostamide, forskolin, and their combination significantly (P < 0.05) increased the percentage of growing (BCB-) oocytes maintained at the germinal vesicle stage. However, only forskolin significantly (P < 0.05) increased the yield and quality of blastocysts derived from BCB- oocytes compared with non-BCB-treated oocytes. We conclude that a short prematuration culture with forskolin may improve the in vitro developmental competency of growing oocytes in ovine. Copyright © 2016 Elsevier Inc. All rights reserved.

  1. From big data to deep insight in developmental science.

    PubMed

    Gilmore, Rick O

    2016-01-01

    The use of the term 'big data' has grown substantially over the past several decades and is now widespread. In this review, I ask what makes data 'big' and what implications the size, density, or complexity of datasets have for the science of human development. A survey of existing datasets illustrates how existing large, complex, multilevel, and multimeasure data can reveal the complexities of developmental processes. At the same time, significant technical, policy, ethics, transparency, cultural, and conceptual issues associated with the use of big data must be addressed. Most big developmental science data are currently hard to find and cumbersome to access, the field lacks a culture of data sharing, and there is no consensus about who owns or should control research data. But, these barriers are dissolving. Developmental researchers are finding new ways to collect, manage, store, share, and enable others to reuse data. This promises a future in which big data can lead to deeper insights about some of the most profound questions in behavioral science. © 2016 The Authors. WIREs Cognitive Science published by Wiley Periodicals, Inc.

  2. Long-Term Growth of Moss in Microfluidic Devices Enables Subcellular Studies in Development.

    PubMed

    Bascom, Carlisle S; Wu, Shu-Zon; Nelson, Katherine; Oakey, John; Bezanilla, Magdalena

    2016-09-01

    Key developmental processes that occur on the subcellular and cellular level or occur in occluded tissues are difficult to access, let alone image and analyze. Recently, culturing living samples within polydimethylsiloxane (PDMS) microfluidic devices has facilitated the study of hard-to-reach developmental events. Here, we show that an early diverging land plant, Physcomitrella patens, can be continuously cultured within PDMS microfluidic chambers. Because the PDMS chambers are bonded to a coverslip, it is possible to image P. patens development at high resolution over long time periods. Using PDMS chambers, we report that wild-type protonemal tissue grows at the same rate as previously reported for growth on solid medium. Using long-term imaging, we highlight key developmental events, demonstrate compatibility with high-resolution confocal microscopy, and obtain growth rates for a slow-growing mutant. By coupling the powerful genetic tools available to P. patens with long-term growth and imaging provided by PDMS microfluidic chambers, we demonstrate the capability to study cellular and subcellular developmental events in plants directly and in real time. © 2016 American Society of Plant Biologists. All rights reserved.

  3. Simple Estimation of the Endolymphatic Volume Ratio after Intravenous Administration of a Single-dose of Gadolinium Contrast

    PubMed Central

    NAGANAWA, Shinji; KANOU, Mai; OHASHI, Toshio; KUNO, Kayao; SONE, Michihiko

    2016-01-01

    Purpose: To evaluate the feasibility of a simple estimation for the endolymphatic volume ratio (endolymph volume/total lymph volume = %ELvolume) from an area ratio obtained from only one slice (%EL1slice) or from three slices (%EL3slices). The %ELvolume, calculated from a time-consuming measurement on all magnetic resonance (MR) slices, was compared to the %EL1slice and the %EL3slices. Methods: In 40 ears of 20 patients with a clinical suspicion of endolymphatic hydrops, MR imaging was performed 4 hours after intravenous administration of a single dose of gadolinium-based contrast material (IV-SD-GBCM). Using previously reported HYDROPS2-Mi2 MR imaging, the %ELvolume values in the cochlea and the vestibule were measured separately by two observers. The correlations between the %EL1slice or the %EL3slices and the %ELvolume values were evaluated. Results: A strong linear correlation was observed between the %ELvolume and the %EL3slices or the %EL1slice in the cochlea. The Pearson correlation coefficient (r) was 0.968 (3 slices) and 0.965 (1 slice) for observer A, and 0.968 (3 slices) and 0.964 (1 slice) for observer B (P < 0.001, for all). A strong linear correlation was also observed between the %ELvolume and the %EL3slices or the %EL1slice in the vestibule. The Pearson correlation coefficient (r) was 0.980 (3 slices) and 0.953 (1 slice) for observer A, and 0.979 (3 slices) and 0.952 (1 slice) for observer B (P < 0.001, for all). The high intra-class correlation coefficients (0.991–0.997) between the endolymph volume ratios by two observers were observed in both the cochlea and the vestibule for values of the %ELvolume, the %EL3slices and the %EL1slice. Conclusion: The %ELvolume might be easily estimated from the %EL3slices or the %EL1slice. PMID:27001396

  4. Simple Estimation of the Endolymphatic Volume Ratio after Intravenous Administration of a Single-dose of Gadolinium Contrast.

    PubMed

    Naganawa, Shinji; Kanou, Mai; Ohashi, Toshio; Kuno, Kayao; Sone, Michihiko

    2016-10-11

    To evaluate the feasibility of a simple estimation for the endolymphatic volume ratio (endolymph volume/total lymph volume = %EL volume ) from an area ratio obtained from only one slice (%EL 1slice ) or from three slices (%EL 3slices ). The %EL volume, calculated from a time-consuming measurement on all magnetic resonance (MR) slices, was compared to the %EL 1slice and the %EL 3slices . In 40 ears of 20 patients with a clinical suspicion of endolymphatic hydrops, MR imaging was performed 4 hours after intravenous administration of a single dose of gadolinium-based contrast material (IV-SD-GBCM). Using previously reported HYDROPS2-Mi2 MR imaging, the %EL volume values in the cochlea and the vestibule were measured separately by two observers. The correlations between the %EL 1slice or the %EL 3slices and the %EL volume values were evaluated. A strong linear correlation was observed between the %EL volume and the %EL 3slices or the %EL 1slice in the cochlea. The Pearson correlation coefficient (r) was 0.968 (3 slices) and 0.965 (1 slice) for observer A, and 0.968 (3 slices) and 0.964 (1 slice) for observer B (P < 0.001, for all). A strong linear correlation was also observed between the %EL volume and the %EL 3slices or the %EL 1slice in the vestibule. The Pearson correlation coefficient (r) was 0.980 (3 slices) and 0.953 (1 slice) for observer A, and 0.979 (3 slices) and 0.952 (1 slice) for observer B (P < 0.001, for all). The high intra-class correlation coefficients (0.991-0.997) between the endolymph volume ratios by two observers were observed in both the cochlea and the vestibule for values of the %EL volume , the %EL 3slices and the %EL 1slice . The %EL volume might be easily estimated from the %EL 3slices or the %EL 1slice .

  5. Developmental consequences of early parenting experiences: self-recognition and self-regulation in three cultural communities.

    PubMed

    Keller, Heidi; Yovsi, Relindis; Borke, Joern; Kärtner, Joscha; Jensen, Henning; Papaligoura, Zaira

    2004-01-01

    This study relates parenting of 3-month-old children to children's self-recognition and self-regulation at 18 to 20 months. As hypothesized, observational data revealed differences in the sociocultural orientations of the 3 cultural samples' parenting styles and in toddlers' development of self-recognition and self-regulation. Children of Cameroonian Nso farmers who experience a proximal parenting style develop self-regulation earlier, children of Greek urban middle-class families who experience a distal parenting style develop self-recognition earlier, and children of Costa Rican middle-class families who experience aspects of both distal and proximal parenting styles fall between the other 2 groups on both self-regulation and self-recognition. Results are discussed with respect to their implications for culturally informed developmental pathways.

  6. A 3D Fibroblast-Epithelium Co-culture Model for Understanding Microenvironmental Role in Branching Morphogenesis of the Mammary Gland.

    PubMed

    Koledova, Zuzana; Lu, Pengfei

    2017-01-01

    The mammary gland consists of numerous tissue compartments, including mammary epithelium, an array of stromal cells, and the extracellular matrix (ECM). Bidirectional interactions between the epithelium and its surrounding stroma are essential for proper mammary gland development and homeostasis, whereas their deregulation leads to developmental abnormalities and cancer. To study the relationships between the epithelium and the stroma, development of models that could recapitulate essential aspects of these interacting systems in vitro has become necessary. Here we describe a three-dimensional (3D) co-culture assay and show that the addition of fibroblasts to mammary organoid cultures promotes the epithelium to undergo branching morphogenesis, thus allowing the role of the stromal microenvironment to be examined in this essential developmental process.

  7. BID Mediates Oxygen-Glucose Deprivation-Induced Neuronal Injury in Organotypic Hippocampal Slice Cultures and Modulates Tissue Inflammation in a Transient Focal Cerebral Ischemia Model without Changing Lesion Volume

    PubMed Central

    Martin, Nellie Anne; Bonner, Helena; Elkjær, Maria Louise; D’Orsi, Beatrice; Chen, Gang; König, Hans Georg; Svensson, Martina; Deierborg, Tomas; Pfeiffer, Shona; Prehn, Jochen H.; Lambertsen, Kate Lykke

    2016-01-01

    The BH3 interacting-domain death agonist (BID) is a pro-apoptotic protein involved in death receptor-induced and mitochondria-mediated apoptosis. Recently, it has also been suggested that BID is involved in the regulation of inflammatory responses in the central nervous system. We found that BID deficiency protected organotypic hippocampal slice cultures in vitro from neuronal injury induced by oxygen-glucose deprivation. In vivo, BID-knockout (KO) mice and wild type (WT) mice were subjected to 60 min of transient middle cerebral artery occlusion (tMCAO) to induce focal cerebral ischemia, and allowed to recover for 24 h. Infarct volumes and functional outcome were assessed and the inflammatory response was evaluated using immunofluorescence, Western blotting, quantitative PCR (qPCR) and Mesoscale multiplex analysis. We observed no difference in the infarct volume or neurological outcome between BID-KO and WT mice. The inflammatory response was reduced by BID deficiency as indicated by a change in microglial/leukocyte response. In conclusion, our data suggest that BID deficiency is neuroprotective in an in vitro model and modulates the inflammatory response to focal cerebral ischemia in vivo. However, this is not translated into a robust neuroprotection in vivo. PMID:26869884

  8. The Microtubule-Associated Protein Doublecortin-Like Regulates the Transport of the Glucocorticoid Receptor in Neuronal Progenitor Cells

    PubMed Central

    Fitzsimons, Carlos P.; Ahmed, Suaad; Wittevrongel, Christiaan F. W.; Schouten, Theo G.; Dijkmans, Thomas F.; Scheenen, Wim J. J. M.; Schaaf, Marcel J. M.; Ronald de Kloet, E.; Vreugdenhil, Erno

    2008-01-01

    In neuronal cells, activated glucocorticoid receptor (GR) translocates to the nucleus guided by the cytoskeleton. However, the detailed mechanisms underlying GR translocation remain unclear. Using gain and loss of function studies, we report here for the first time that the microtubule-associated protein doublecortin-like (DCL) controls GR translocation to the nucleus. DCL overexpression in COS-1 cells, neuroblastoma cells, and rat hippocampus organotypic slice cultures impaired GR translocation and decreased GR-dependent transcriptional activity, measured by a specific reporter gene assay, in COS-1 cells. Moreover, DCL and GR directly interact on microtubule bundles formed by DCL overexpression. A C-terminal truncated DCL with conserved microtubule-bundling activity did not influence GR translocation. In N1E-115 mouse neuroblastoma cells and neuronal progenitor cells in rat hippocampus organotypic slice cultures, laser-scanning confocal microscopy showed colabeling of endogenously expressed DCL and GR. In these systems, RNA-interference-mediated DCL knockdown hampered GR translocation. Thus, we conclude that DCL expression is tightly regulated to adequately control GR transport. Because DCL is primarily expressed in neuronal progenitor cells, our results introduce this microtubule-associated protein as a new modulator of GR signaling in this cell type and suggest the existence of cell-specific mechanisms regulating GR translocation to the nucleus. PMID:17975023

  9. Time-lapse imaging reveals highly dynamic structural maturation of postnatally born dentate granule cells in organotypic entorhino-hippocampal slice cultures

    PubMed Central

    Radic, Tijana; Jungenitz, Tassilo; Singer, Mathias; Beining, Marcel; Cuntz, Hermann; Vlachos, Andreas; Deller, Thomas; Schwarzacher, Stephan W.

    2017-01-01

    Neurogenesis of hippocampal granule cells (GCs) persists throughout mammalian life and is important for learning and memory. How newborn GCs differentiate and mature into an existing circuit during this time period is not yet fully understood. We established a method to visualize postnatally generated GCs in organotypic entorhino-hippocampal slice cultures (OTCs) using retroviral (RV) GFP-labeling and performed time-lapse imaging to study their morphological development in vitro. Using anterograde tracing we could, furthermore, demonstrate that the postnatally generated GCs in OTCs, similar to adult born GCs, grow into an existing entorhino-dentate circuitry. RV-labeled GCs were identified and individual cells were followed for up to four weeks post injection. Postnatally born GCs exhibited highly dynamic structural changes, including dendritic growth spurts but also retraction of dendrites and phases of dendritic stabilization. In contrast, older, presumably prenatally born GCs labeled with an adeno-associated virus (AAV), were far less dynamic. We propose that the high degree of structural flexibility seen in our preparations is necessary for the integration of newborn granule cells into an already existing neuronal circuit of the dentate gyrus in which they have to compete for entorhinal input with cells generated and integrated earlier. PMID:28256620

  10. Industrial application of an antilisterial strain of Lactobacillus sakei as a protective culture and its effect on the sensory acceptability of cooked, sliced, vacuum-packaged meats.

    PubMed

    Bredholt, S; Nesbakken, T; Holck, A

    2001-06-15

    The application of a protective lactic acid bacterium (LAB) during the commercial production of cooked meat products is described. The LAB, a strain of Lactobacillus sakei, was previously isolated from cooked ham and inhibited growth of Listeria monocytogenes and Escherichia coli O157:H7 in this product. L. sakei was applied to the cooked products at a concentration of 10(5)-10(6) cfu/g immediately before slicing and vacuum-packaging using a hand-operated spraying bottle. The LAB strain inhibited growth of 10(3) cfu/g of a cocktail of three rifampicin resistant mutant L. monocytogenes strains both at 8 degrees C and 4 degrees C. Consumer acceptance tests of cooked ham and of servelat sausage, a Norwegian non-fermented cooked meat sausage, showed that control and inoculated products were equally acceptable. The products were still acceptable after storage for 28 days at 4 degrees C and, after opening the packages, for a further 5 days at 4 degrees C. The findings presented here confirm that the L. sakei strain is suitable for use as a protective culture and may technically easily be implemented in the commercial production of cooked meat products.

  11. Thin-Slice Measurement of Wisdom

    PubMed Central

    Hu, Chao S.; Ferrari, Michel; Wang, Qiandong; Woodruff, Earl

    2017-01-01

    Objective Measurement of Wisdom within a short period of time is vital for both the public interest (e.g., understanding a presidential election) and research (e.g., testing factors that facilitate wisdom development). A measurement of emotion associated with wisdom would be especially informative; therefore, a novel Thin-Slice measurement of wisdom was developed based on the Berlin Paradigm. For about 2 min, participants imagined the lens of a camera as the eyes of their friend/teacher whom they advised about a life dilemma. Verbal response and facial expression were both recorded by a camera: verbal responses were then rated on both the Berlin Wisdom criteria and newly developed Chinese wisdom criteria; facial expressions were analyzed by the software iMotion FACET module. Results showed acceptable inter-rater and inter-item reliability for this novel paradigm. Moreover, both wisdom ratings were not significantly correlated with Social desirability, and the Berlin wisdom rating was significantly negatively correlated with Neuroticism; feeling of surprise was significantly positively correlated with both wisdom criteria ratings. Our results provide the first evidence of this Thin-slice Wisdom Paradigm’s reliability, its immunity to social desirability, and its validity for assessing candidates’ wisdom within a short timeframe. Although still awaiting further development, this novel Paradigm contributes to an emerging Universal Wisdom Paradigm applicable across cultures. PMID:28861016

  12. Bovine oocyte meiotic inhibition before in vitro maturation and its value to in vitro embryo production: does it improve developmental competence?

    PubMed

    Bilodeau-Goeseels, S

    2012-08-01

    The efficiency of bovine in vitro embryo production has remained low despite extensive effort to understand the effects of culture conditions, media composition and supplementation. As bovine oocytes resume meiosis spontaneously when cultured, it was hypothesized that preventing meiosis in vitro before in vitro maturation (IVM) and in vitro fertilization (IVF) would allow more oocytes to acquire developmental competence. This article reviews some of the factors involved in meiotic arrest as well as the effects of meiotic inhibition before IVM on bovine oocytes developmental competence following IVF. Follicular components and cAMP-elevating agents can delay or inhibit meiosis in various proportions of oocytes; however, few studies have examined their effects on development following IVM and IVF because they are not practical (follicular components) or have a transient effect on meiosis (cAMP-elevating agents). Protein synthesis or phosphorylation inhibition prevented meiosis in high percentages of oocytes; however, these non-specific inhibitions led to lower developmental competence compared with non-arrested oocytes. Maturation promoting factor (MPF) inhibition with specific inhibitors has been examined in several studies. Despite faster maturation following removal from inhibition and some structural damage to the oocytes, MPF inhibition generally led to blastocyst rates similar to control, non-arrested oocytes. Future work will involve evaluating the effects on arrested oocytes of molecules that can improve developmental competence in non-arrested oocytes. It is also anticipated that new IVM systems that take into consideration new knowledge of the mechanisms involved in the control of meiosis will be developed. Moreover, global gene expression analysis studies will also provide clues to the culture conditions required for optimal expression of developmental competence. © Her Majesty the Queen in Right of Canada 2011. Reproduced with the permission of the Minister of Agriculture and Agri-Food Canada.

  13. Performance of South African children on the Communication and Symbolic Behavior Scales-Developmental Profile (CSBS DP).

    PubMed

    Chambers, Nola; Stronach, Sheri T; Wetherby, Amy M

    2016-05-01

    Substantial development in social communication skills occurs in the first two years of life. Growth should be evident in sharing emotion and eye gaze; rate of communication, communicating for a variety of functions; using gestures, sounds and words; understanding language, and using functional and pretend actions with objects in play. A delay in these early social communication skills may be the first sign of a developmental delay in young children in nearly all categories of disabilities-including specific language impairment, autism spectrum disorder, HIV/AIDS, lack of environmental stimulation or institutionalization, and global developmental delays-and early detection of these delays is critical for enrolment in appropriate early intervention services. No standardized tests of early social communication skills exist for very young children in South Africa (SA). An existing evaluation tool that has the potential to be culturally fair for children from cultural backgrounds different to the standardization group is the Communication and Symbolic Behaviour Scales-Developmental Profile (CSBS DP). This study aimed to document the performance of a group of English-speaking SA children ranging in age from 12 to 24 months on the CSBS DP and to compare this performance with the original standardization sample. Sixty-seven English-speaking SA children from a range of cultural and linguistic backgrounds were assessed on the CSBS DP Behaviour Sample. Group scores were compared with the original standardization sample using inferential statistics. The results provide preliminary support for the suitability and validity of the face-to-face Behaviour Sample as a measure of early social communication skills in this sample of English-speaking SA children from a range of cultural groups between 12 and 24 months of age. While further research in the SA population is needed, these findings are a first step towards validating a culturally appropriate measure for early detection of social communication delays in a sample of SA toddlers. © 2015 Royal College of Speech and Language Therapists.

  14. Over a century of neuron culture: from the hanging drop to microfluidic devices.

    PubMed

    Millet, Larry J; Gillette, Martha U

    2012-12-01

    The brain is the most intricate, energetically active, and plastic organ in the body. These features extend to its cellular elements, the neurons and glia. Understanding neurons, or nerve cells, at the cellular and molecular levels is the cornerstone of modern neuroscience. The complexities of neuron structure and function require unusual methods of culture to determine how aberrations in or between cells give rise to brain dysfunction and disease. Here we review the methods that have emerged over the past century for culturing neurons in vitro, from the landmark finding by Harrison (1910) - that neurons can be cultured outside the body - to studies utilizing culture vessels, micro-islands, Campenot and brain slice chambers, and microfluidic technologies. We conclude with future prospects for neuronal culture and considerations for advancement. We anticipate that continued innovation in culture methods will enhance design capabilities for temporal control of media and reagents (chemotemporal control) within sub-cellular environments of three-dimensional fluidic spaces (microfluidic devices) and materials (e.g., hydrogels). They will enable new insights into the complexities of neuronal development and pathology.

  15. Over a Century of Neuron Culture: From the Hanging Drop to Microfluidic Devices

    PubMed Central

    Millet, Larry J.; Gillette, Martha U.

    2012-01-01

    The brain is the most intricate, energetically active, and plastic organ in the body. These features extend to its cellular elements, the neurons and glia. Understanding neurons, or nerve cells, at the cellular and molecular levels is the cornerstone of modern neuroscience. The complexities of neuron structure and function require unusual methods of culture to determine how aberrations in or between cells give rise to brain dysfunction and disease. Here we review the methods that have emerged over the past century for culturing neurons in vitro, from the landmark finding by Harrison (1910) — that neurons can be cultured outside the body — to studies utilizing culture vessels, micro-islands, Campenot and brain slice chambers, and microfluidic technologies. We conclude with future prospects for neuronal culture and considerations for advancement. We anticipate that continued innovation in culture methods will enhance design capabilities for temporal control of media and reagents (chemotemporal control) within sub-cellular environments of three-dimensional fluidic spaces (microfluidic devices) and materials (e.g., hydrogels). They will enable new insights into the complexities of neuronal development and pathology. PMID:23239951

  16. Zscan4 restores the developmental potency of embryonic stem cells

    PubMed Central

    Amano, Tomokazu; Hirata, Tetsuya; Falco, Geppino; Monti, Manuela; Sharova, Lioudmila V.; Amano, Misa; Sheer, Sarah; Hoang, Hien G.; Piao, Yulan; Stagg, Carole A.; Yamamizu, Kohei; Akiyama, Tomohiko; Ko, Minoru S.H.

    2013-01-01

    The developmental potency of mouse embryonic stem (ES) cells, which is the ability to contribute to a whole embryo is known to deteriorate during long-term cell culture. Previously we have shown that ES cells oscillate between Zscan4- and Zscan4+ states, and the transient activation of Zscan4 is required for the maintenance of telomeres and genome stability of ES cells. Here we show that increasing the frequency of Zscan4 activation in mouse ES cells restores and maintains their developmental potency in long-term cell culture. Injection of a single ES cell with such increased potency into a tetraploid blastocyst gives rise to an entire embryo with a higher success rate. These results not only provide a means to rejuvenate ES cells by manipulating Zscan4 expression, but also indicate the active roles of Zscan4 in the long-term maintenance of ES cell potency. PMID:23739662

  17. Bringing together local culture and rural development: findings from Ireland, Pennsylvania and Alaska

    Treesearch

    M.A. Brennan; Courtney G. Flint; A.E. Luloff

    2009-01-01

    The developmental trajectories of communities are routinely explained by reference to economic history, human capital deficits, or the structure of local labour markets. The role of local culture in understanding community development or in interpreting empirical research has received less attention. We believe culture plays an important independent role in shaping...

  18. Child Development in Cultural Contexts: Implications of Cultural Psychology for Early Childhood Teacher Education

    ERIC Educational Resources Information Center

    Lee, Kyunghwa; Johnson, Amy S.

    2007-01-01

    In this article we argue that early childhood educators, under the influence of last century's grand universal theories of child development, have not been attentive enough to the centrality of culture in children's development. We discuss how the exploration of contemporary developmental perspectives is critical to the field and illustrate…

  19. Dose–response analysis of phthalate effects on gene expression in rat whole embryo culture

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

    Robinson, Joshua F.; Department of Toxicogenomics, Maastricht University, Maastricht; Verhoef, Aart

    2012-10-01

    The rat postimplantation whole embryo culture (WEC) model serves as a potential screening tool for developmental toxicity. In this model, cultured rat embryos are exposed during early embryogenesis and evaluated for morphological effects. The integration of molecular-based markers may lead to improved objectivity, sensitivity and predictability of WEC in assessing developmental toxic properties of compounds. In this study, we investigated the concentration-dependent effects of two phthalates differing in potency, mono(2-ethylhexyl) phthalate (MEHP) and monomethyl phthalate (MMP, less toxic), on the transcriptome in WEC to examine gene expression in relation with dysmorphogenesis. MEHP was more potent than MMP in inducing genemore » expression changes as well as changes on morphology. MEHP induced significant enrichment of cholesterol/lipid/steroid (CLS) metabolism and apoptosis pathways which was associated with developmental toxicity. Regulation of genes within CLS metabolism pathways represented the most sensitive markers of MEHP exposure, more sensitive than classical morphological endpoints. As shown in direct comparisons with toxicogenomic in vivo studies, alterations in the regulation of CLS metabolism pathways has been previously identified to be associated with developmental toxicity due to phthalate exposure in utero. Our results support the application of WEC as a model to examine relative phthalate potency through gene expression and morphological responses. Additionally, our results further define the applicability domain of the WEC model for developmental toxicological investigations. -- Highlights: ► We examine the effect of two phthalates on gene expression and morphology in WEC. ► MEHP is more potent than MMP in inducing gene expression changes and dysmorphogenesis. ► MEHP significantly disrupts cholesterol metabolism pathways in a dose-dependent manner. ► Specific phthalate-related mechanisms in WEC are relevant to mechanisms in vivo.« less

  20. A Vygotskian Approach to Heterogeneous Communication and Multi/Cultural Literacy: Commentary on David Kellogg's "Taking Uptaking up, or, a Deconstructionist "Ontology of Difference" and a Developmental One"

    ERIC Educational Resources Information Center

    Hwang, SungWon

    2009-01-01

    In this commentary, I review Kellogg's comments on a recent editorial in the journal "Mind, Culture, and Activity" (Roth, 2008). Concerning Kellogg's code-switching model for learning language, I present and exemplify a dialectic problem of multi/cultural literacy: the first articulation that crosses the boundaries of cultures and languages…

  1. Environment, Biology, and Culture: Implications for Adolescent Development.

    ERIC Educational Resources Information Center

    Zahn-Waxler, Carolyn

    1996-01-01

    Introduces this special theme issue examining the roles of socialization, biology, and culture as they affect adaptive and maladaptive developmental outcomes. Problems of adolescence addressed include antisocial behavior, depressive symptoms, substance abuse, low achievement, and eating problems. Considers factors implicated in successful…

  2. Ventral tegmental area/substantia nigra and prefrontal cortex rodent organotypic brain slices as an integrated model to study the cellular changes induced by oxygen/glucose deprivation and reperfusion: effect of neuroprotective agents.

    PubMed

    Colombo, Laura; Parravicini, Chiara; Lecca, Davide; Dossi, Elena; Heine, Claudia; Cimino, Mauro; Wanke, Enzo; Illes, Peter; Franke, Heike; Abbracchio, Maria P

    2014-01-01

    Unveiling the roles of distinct cell types in brain response to insults is a partially unsolved challenge and a key issue for new neuroreparative approaches. In vivo models are not able to dissect the contribution of residential microglia and infiltrating blood-borne monocytes/macrophages, which are fundamentally undistinguishable; conversely, cultured cells lack original tissue anatomical and functional complexity, which profoundly alters reactivity. Here, we tested whether rodent organotypic co-cultures from mesencephalic ventral tegmental area/substantia nigra and prefrontal cortex (VTA/SN-PFC) represent a suitable model to study changes induced by oxygen/glucose deprivation and reperfusion (OGD/R). OGD/R induced cytotoxicity to both VTA/SN and PFC slices, with higher VTA/SN susceptibility. Neurons were highly affected, with astrocytes and oligodendrocytes undergoing very mild damage. Marked reactive astrogliosis was also evident. Notably, OGD/R triggered the activation of CD68-expressing microglia and increased expression of Ym1 and Arg1, two markers of "alternatively" activated beneficial microglia. Treatment with two well-known neuroprotective drugs, the anticonvulsant agent valproic acid and the purinergic P2-antagonist PPADS, prevented neuronal damage. Thus, VTA/SN-PFC cultures are an integrated model to investigate OGD/R-induced effects on distinct cells and easily screen neuroprotective agents. The model is particularly adequate to dissect the microglia phenotypic shift in the lack of a functional vascular compartment. Copyright © 2014 Elsevier Ltd. All rights reserved.

  3. Teaching the Standards Is Developmentally Appropriate Practice: Strategies for Incorporating the Sociopolitical Dimension of DAP in Early Childhood Teaching

    ERIC Educational Resources Information Center

    Goldstein, Lisa S.

    2008-01-01

    Many early childhood practitioners in the U.S. are experiencing tension between their desire to offer students developmentally appropriate learning experiences and their obligation to teach the academic knowledge and skills mandated by their states. However, careful examination of the DAP guidelines' definition of culturally appropriate practice…

  4. Interventions for Children with Pervasive Developmental Disorders in Low and Middle Income Countries

    ERIC Educational Resources Information Center

    Hastings, Richard P.; Robertson, Janet; Yasamy, M. T.

    2012-01-01

    Background: Although interventions for children with pervasive developmental disorders (PDD) have been the focus of research effort and evidence reviews in many Western countries, this evidence has not been assessed in the context of low- and middle-income (LAMI) countries especially in terms of the fit with different cultures and resources.…

  5. Developmentally Appropriate Technology in Early Childhood (DATEC) in Botswana: In-Service Teachers' Perspectives

    ERIC Educational Resources Information Center

    Bose, Kabita

    2009-01-01

    Developmentally Appropriate Technology in Early Childhood (DATEC) aims to identify the most appropriate applications of Information and Communication Technology to support the development of children under eight years of age. Botswana has a unique spread of population density and deep-rooted socio-cultural values. There is a need to address the…

  6. Was Pre-Modern Man a Child? The Quintessence of the Psychometric and Developmental Approaches

    ERIC Educational Resources Information Center

    Oesterdiekhoff, Georg W.

    2012-01-01

    The essay integrates the psychometric intelligence approach with the cognitive-developmental approach or the stage theory erected by Piaget and his disciples. The latter led to Piagetian Cross-Cultural Psychology and the accumulation of an immense body of data. It shows that different IQ levels are indicative of the peculiar stages of cognitive…

  7. Cross-Cultural Adaptation of a Pre-School Screening Instrument: Comparison of Korean and US Populations

    ERIC Educational Resources Information Center

    Heo, K. H.; Squires, J.; Yovanoff, P.

    2008-01-01

    Background: Accurate and efficient developmental screening measures are critical for early identification of developmental problems; however, few reliable and valid tests are available in Korea as well as other countries outside the USA. The Ages and Stages Questionnaires (ASQ) was chosen for study with young children in Korea. Methods: The ASQ…

  8. Cross-Cultural Validity of the Social Communication Questionnaire for Adults with Intellectual Developmental Disorder

    ERIC Educational Resources Information Center

    Sappok, Tanja; Brooks, Whitney; Heinrich, Manuel; McCarthy, Jane; Underwood, Lisa

    2017-01-01

    Diagnosing Autism Spectrum Disorders (ASD) is important throughout the lifespan. The objective was to investigate the transcultural diagnostic validity of the Social Communication Questionnaire (SCQ) in a clinical sample of 451 adults with Intellectual Developmental Disorder (IDD) with and without ASD in Germany, the U.S.A. and Great Britain.…

  9. Child Development and Cultural Diversity.

    ERIC Educational Resources Information Center

    Caldwell, Bettye M.

    1983-01-01

    Each child is to some extent like all children, to some extent like some children, and to some extent like no other child. There are at least three sets of universals that characterize children: (1) they have the same needs and rights; (2) they go through the same developmental stages; and (3) they have essentially the same developmental goals,…

  10. Large-scale gene expression profiling data for the model moss Physcomitrella patens aid understanding of developmental progression, culture and stress conditions.

    PubMed

    Hiss, Manuel; Laule, Oliver; Meskauskiene, Rasa M; Arif, Muhammad A; Decker, Eva L; Erxleben, Anika; Frank, Wolfgang; Hanke, Sebastian T; Lang, Daniel; Martin, Anja; Neu, Christina; Reski, Ralf; Richardt, Sandra; Schallenberg-Rüdinger, Mareike; Szövényi, Peter; Tiko, Theodhor; Wiedemann, Gertrud; Wolf, Luise; Zimmermann, Philip; Rensing, Stefan A

    2014-08-01

    The moss Physcomitrella patens is an important model organism for studying plant evolution, development, physiology and biotechnology. Here we have generated microarray gene expression data covering the principal developmental stages, culture forms and some environmental/stress conditions. Example analyses of developmental stages and growth conditions as well as abiotic stress treatments demonstrate that (i) growth stage is dominant over culture conditions, (ii) liquid culture is not stressful for the plant, (iii) low pH might aid protoplastation by reduced expression of cell wall structure genes, (iv) largely the same gene pool mediates response to dehydration and rehydration, and (v) AP2/EREBP transcription factors play important roles in stress response reactions. With regard to the AP2 gene family, phylogenetic analysis and comparison with Arabidopsis thaliana shows commonalities as well as uniquely expressed family members under drought, light perturbations and protoplastation. Gene expression profiles for P. patens are available for the scientific community via the easy-to-use tool at https://www.genevestigator.com. By providing large-scale expression profiles, the usability of this model organism is further enhanced, for example by enabling selection of control genes for quantitative real-time PCR. Now, gene expression levels across a broad range of conditions can be accessed online for P. patens. © 2014 The Authors The Plant Journal © 2014 John Wiley & Sons Ltd.

  11. In vitro culture medium (IVC) supplementation with sericin improves developmental competence of ovine zygotes.

    PubMed

    Aghaz, Faranak; Hajarian, Hadi; KaramiShabankareh, Hamed

    2016-03-01

    This study was carried out to investigate the effects of supplementation of potassium simplex optimized medium (KSOM-aa) with various sericin concentrations (0, 0.1, 0.5, 1 and 2.5%) on ovine zygotes. The results indicate that the supplementation of oocyte in vitro culture medium with optimal concentration of sericin (0.1 and 0.5%) may have beneficial effects on developmental competence of in vitro-derived ovine embryos. Copyright © 2015 Society for Biology of Reproduction & the Institute of Animal Reproduction and Food Research of Polish Academy of Sciences in Olsztyn. Published by Elsevier Urban & Partner Sp. z o.o. All rights reserved.

  12. A tribute to Dr. Gordon Hisashi Sato (December 24, 1927-March 31, 2017).

    PubMed

    Sato, J Denry; Okamoto, Tetsuji; Barnes, David; Hayashi, Jun; Serrero, Ginette; McKeehan, Wallace L

    2018-03-01

    Gordon H. Sato, an innovator in mammalian tissue culture and integrated cellular physiology, passed away in 2017. In tribute to Dr. Sato, In Vitro Cellular and Developmental Biology-Animal presents a collection of invited remembrances from six colleagues whose associations with Dr. Sato spanned more than 40 years. Dr. Sato was a past president of the Tissue Culture Association (now the Society for In Vitro Biology), editor-in-chief of In Vitro Cellular and Developmental Biology (1987-1991), and the recipient of the lifetime achievement award from the Society for In Vitro Biology (2002). He was elected to the US National Academy of Sciences in 1984.

  13. Cross-cultural similarities and differences in person-body reasoning: experimental evidence from the United Kingdom and Brazilian Amazon.

    PubMed

    Cohen, Emma; Burdett, Emily; Knight, Nicola; Barrett, Justin

    2011-01-01

    We report the results of a cross-cultural investigation of person-body reasoning in the United Kingdom and northern Brazilian Amazon (Marajó Island). The study provides evidence that directly bears upon divergent theoretical claims in cognitive psychology and anthropology, respectively, on the cognitive origins and cross-cultural incidence of mind-body dualism. In a novel reasoning task, we found that participants across the two sample populations parsed a wide range of capacities similarly in terms of the capacities' perceived anchoring to bodily function. Patterns of reasoning concerning the respective roles of physical and biological properties in sustaining various capacities did vary between sample populations, however. Further, the data challenge prior ad-hoc categorizations in the empirical literature on the developmental origins of and cognitive constraints on psycho-physical reasoning (e.g., in afterlife concepts). We suggest cross-culturally validated categories of "Body Dependent" and "Body Independent" items for future developmental and cross-cultural research in this emerging area. Copyright © 2011 Cognitive Science Society, Inc.

  14. Psychological essentialism among Korean children and adults: a modified replication study.

    PubMed

    Kim, Geunyoung

    2013-01-01

    Developmental psychologists have shown interest in the development of psychological essentialism among children; that is, a belief that certain psychological characteristics (such as personality) are relatively stable and unchanging. Although previous studies have shown that children are essentialistic about human traits, and the coherence among various essentialism dimensions increases with age, moderating cultural factors in the development of essentialism, especially among Asian children, have received little attention. Using the methodologies of Gelman, Heyman, and Legare (2007), levels of psychological essentialism among Korean children and adults were measured, and compared with the original US data. Results demonstrated cross-cultural similarity in the development of coherence in essentialistic thought, and difference in the level of essentialism among adult participants. The present findings imply that different cultural values between Asians and Westerners can play a role in the developmental trajectory of psychological essentialism.

  15. Music, empathy and cultural understanding: The need for developmental research. Comment on "Music, empathy and cultural understanding" by E. Clarke et al.

    NASA Astrophysics Data System (ADS)

    Rabinowitch, Tal-Chen

    2015-12-01

    Clarke, DeNora and Vuoskoski have carried out a formidable task of preparing a profound and encompassing review [3] that brings together two highly complex and multifaceted concepts, empathy and music, as well as a specific aspect of empathy that is highly relevant to society, cultural understanding. They have done an extraordinary service in synthesizing the growing, but still highly fragmented body of work in this area. At the heart of this review lies an intricate model that the authors develop, which accounts for a variety of mechanisms and cognitive processes underlying musical empathic engagement. In what follows I would like to first point out what I think is unique about this model. Then, I will briefly describe the need for including in any such model a developmental angle.

  16. Confucian virtues and Chinese adolescent development: a conceptual review.

    PubMed

    Shek, Daniel T L; Yu, Lu; Fu, Xiao

    2013-01-01

    Despite the fact that different Chinese communities have already undergone industrialization and urbanization, Confucian virtues are still regarded as developmental ideals in Chinese culture. Unfortunately, while Confucian virtues are commonly discussed under Chinese philosophies, they are rarely examined in the context of developmental research. In this paper, several key Confucian virtues are discussed, including loyalty ("zhong"), filial piety ("xiao"), benevolence ("ren"), affection ("ai"), trustworthiness ("xin"), righteousness ("yi"), harmony ("he"), peace ("ping"), propriety ("li"), wisdom ("zhi"), integrity ("lian") and shame ("chi"). These Chinese traditional virtues are also linked to the concepts of character strengths and positive youth development constructs highlighted in Western culture. It is argued that Confucian virtues provide an indigenous conceptual framework to understand character strengths and positive youth development in Chinese culture. Furthermore, when service leadership is considered in Chinese contexts, these virtues should be regarded as important cornerstones.

  17. Integration of culture and biology in human development.

    PubMed

    Mistry, Jayanthi

    2013-01-01

    The challenge of integrating biology and culture is addressed in this chapter by emphasizing human development as involving mutually constitutive, embodied, and epigenetic processes. Heuristically rich constructs extrapolated from cultural psychology and developmental science, such as embodiment, action, and activity, are presented as promising approaches to the integration of cultural and biology in human development. These theoretical notions are applied to frame the nascent field of cultural neuroscience as representing this integration of culture and biology. Current empirical research in cultural neuroscience is then synthesized to illustrate emerging trends in this body of literature that examine the integration of biology and culture.

  18. Cognitive consistency and math-gender stereotypes in Singaporean children.

    PubMed

    Cvencek, Dario; Meltzoff, Andrew N; Kapur, Manu

    2014-01-01

    In social psychology, cognitive consistency is a powerful principle for organizing psychological concepts. There have been few tests of cognitive consistency in children and no research about cognitive consistency in children from Asian cultures, who pose an interesting developmental case. A sample of 172 Singaporean elementary school children completed implicit and explicit measures of math-gender stereotype (male=math), gender identity (me=male), and math self-concept (me=math). Results showed strong evidence for cognitive consistency; the strength of children's math-gender stereotypes, together with their gender identity, significantly predicted their math self-concepts. Cognitive consistency may be culturally universal and a key mechanism for developmental change in social cognition. We also discovered that Singaporean children's math-gender stereotypes increased as a function of age and that boys identified with math more strongly than did girls despite Singaporean girls' excelling in math. The results reveal both cultural universals and cultural variation in developing social cognition. Copyright © 2013 Elsevier Inc. All rights reserved.

  19. Genetically Targeted All-Optical Electrophysiology with a Transgenic Cre-Dependent Optopatch Mouse

    PubMed Central

    Lou, Shan; Adam, Yoav; Weinstein, Eli N.; Williams, Erika; Williams, Katherine; Parot, Vicente; Kavokine, Nikita; Liberles, Stephen; Madisen, Linda; Zeng, Hongkui

    2016-01-01

    Recent advances in optogenetics have enabled simultaneous optical perturbation and optical readout of membrane potential in diverse cell types. Here, we develop and characterize a Cre-dependent transgenic Optopatch2 mouse line that we call Floxopatch. The animals expressed a blue-shifted channelrhodopsin, CheRiff, and a near infrared Archaerhodopsin-derived voltage indicator, QuasAr2, via targeted knock-in at the rosa26 locus. In Optopatch-expressing animals, we tested for overall health, genetically targeted expression, and function of the optogenetic components. In offspring of Floxopatch mice crossed with a variety of Cre driver lines, we observed spontaneous and optically evoked activity in vitro in acute brain slices and in vivo in somatosensory ganglia. Cell-type-specific expression allowed classification and characterization of neuronal subtypes based on their firing patterns. The Floxopatch mouse line is a useful tool for fast and sensitive characterization of neural activity in genetically specified cell types in intact tissue. SIGNIFICANCE STATEMENT Optical recordings of neural activity offer the promise of rapid and spatially resolved mapping of neural function. Calcium imaging has been widely applied in this mode, but is insensitive to the details of action potential waveforms and subthreshold events. Simultaneous optical perturbation and optical readout of single-cell electrical activity (“Optopatch”) has been demonstrated in cultured neurons and in organotypic brain slices, but not in acute brain slices or in vivo. Here, we describe a transgenic mouse in which expression of Optopatch constructs is controlled by the Cre-recombinase enzyme. This animal enables fast and robust optical measurements of single-cell electrical excitability in acute brain slices and in somatosensory ganglia in vivo, opening the door to rapid optical mapping of neuronal excitability. PMID:27798186

  20. Comparison of the mouse Embryonic Stem cell Test, the rat Whole Embryo Culture and the Zebrafish Embryotoxicity Test as alternative methods for developmental toxicity testing of six 1,2,4-triazoles

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

    Jong, Esther de, E-mail: Esther.de.Jong@rivm.nl; Laboratory for Health Protection Research, National Institute for Public Health and the Environment; Barenys, Marta

    2011-06-01

    The relatively high experimental animal use in developmental toxicity testing has stimulated the search for alternatives that are less animal intensive. Three widely studied alternative assays are the mouse Embryonic Stem cell Test (EST), the Zebrafish Embryotoxicity Test (ZET) and the rat postimplantation Whole Embryo Culture (WEC). The goal of this study was to determine their efficacy in assessing the relative developmental toxicity of six 1,2,4-triazole compounds, flusilazole, hexaconazole, cyproconazole, triadimefon, myclobutanil and triticonazole. For this purpose, we analyzed effects and relative potencies of the compounds in and among the alternative assays and compared the findings to their known inmore » vivo developmental toxicity. Triazoles are antifungal agents used in agriculture and medicine, some of which are known to induce craniofacial and limb abnormalities in rodents. The WEC showed a general pattern of teratogenic effects, typical of exposure to triazoles, mainly consisting of reduction and fusion of the first and second branchial arches, which are in accordance with the craniofacial malformations reported after in vivo exposure. In the EST all triazole compounds inhibited cardiomyocyte differentiation concentration-dependently. Overall, the ZET gave the best correlation with the relative in vivo developmental toxicities of the tested compounds, closely followed by the EST. The relative potencies observed in the WEC showed the lowest correlation with the in vivo developmental toxicity data. These differences in the efficacy between the test systems might be due to differences in compound kinetics, in developmental stages represented and in the relative complexity of the alternative assays.« less

  1. Comparison of the mouse Embryonic Stem cell Test, the rat Whole Embryo Culture and the Zebrafish Embryotoxicity Test as alternative methods for developmental toxicity testing of six 1,2,4-triazoles.

    PubMed

    de Jong, Esther; Barenys, Marta; Hermsen, Sanne A B; Verhoef, Aart; Ossendorp, Bernadette C; Bessems, Jos G M; Piersma, Aldert H

    2011-06-01

    The relatively high experimental animal use in developmental toxicity testing has stimulated the search for alternatives that are less animal intensive. Three widely studied alternative assays are the mouse Embryonic Stem cell Test (EST), the Zebrafish Embryotoxicity Test (ZET) and the rat postimplantation Whole Embryo Culture (WEC). The goal of this study was to determine their efficacy in assessing the relative developmental toxicity of six 1,2,4-triazole compounds,(1) flusilazole, hexaconazole, cyproconazole, triadimefon, myclobutanil and triticonazole. For this purpose, we analyzed effects and relative potencies of the compounds in and among the alternative assays and compared the findings to their known in vivo developmental toxicity. Triazoles are antifungal agents used in agriculture and medicine, some of which are known to induce craniofacial and limb abnormalities in rodents. The WEC showed a general pattern of teratogenic effects, typical of exposure to triazoles, mainly consisting of reduction and fusion of the first and second branchial arches, which are in accordance with the craniofacial malformations reported after in vivo exposure. In the EST all triazole compounds inhibited cardiomyocyte differentiation concentration-dependently. Overall, the ZET gave the best correlation with the relative in vivo developmental toxicities of the tested compounds, closely followed by the EST. The relative potencies observed in the WEC showed the lowest correlation with the in vivo developmental toxicity data. These differences in the efficacy between the test systems might be due to differences in compound kinetics, in developmental stages represented and in the relative complexity of the alternative assays. Copyright © 2011 Elsevier Inc. All rights reserved.

  2. Integration of technologies for hepatic tissue engineering.

    PubMed

    Nahmias, Yaakov; Berthiaume, Francois; Yarmush, Martin L

    2007-01-01

    The liver is the largest internal organ in the body, responsible for over 500 metabolic, regulatory, and immune functions. Loss of liver function leads to liver failure which causes over 25,000 deaths/year in the United States. Efforts in the field of hepatic tissue engineering include the design of bioartificial liver systems to prolong patient's lives during liver failure, for drug toxicity screening and for the study of liver regeneration, ischemia/reperfusion injury, fibrosis, viral infection, and inflammation. This chapter will overview the current state-of-the-art in hepatology including isolated perfused liver, culture of liver slices and tissue explants, hepatocyte culture on collagen "sandwich" and spheroids, coculture of hepatocytes with non-parenchymal cells, and the integration of these culture techniques with microfluidics and reactor design. This work will discuss the role of oxygen and medium composition in hepatocyte culture and present promising new technologies for hepatocyte proliferation and function. We will also discuss liver development, architecture, and function as they relate to these culture techniques. Finally, we will review current opportunities and major challenges in integrating cell culture, bioreactor design, and microtechnology to develop new systems for novel applications.

  3. Clostridium perfringens epsilon toxin targets granule cells in the mouse cerebellum and stimulates glutamate release.

    PubMed

    Lonchamp, Etienne; Dupont, Jean-Luc; Wioland, Laetitia; Courjaret, Raphaël; Mbebi-Liegeois, Corinne; Jover, Emmanuel; Doussau, Frédéric; Popoff, Michel R; Bossu, Jean-Louis; de Barry, Jean; Poulain, Bernard

    2010-09-30

    Epsilon toxin (ET) produced by C. perfringens types B and D is a highly potent pore-forming toxin. ET-intoxicated animals express severe neurological disorders that are thought to result from the formation of vasogenic brain edemas and indirect neuronal excitotoxicity. The cerebellum is a predilection site for ET damage. ET has been proposed to bind to glial cells such as astrocytes and oligodendrocytes. However, the possibility that ET binds and attacks the neurons remains an open question. Using specific anti-ET mouse polyclonal antibodies and mouse brain slices preincubated with ET, we found that several brain structures were labeled, the cerebellum being a prominent one. In cerebellar slices, we analyzed the co-staining of ET with specific cell markers, and found that ET binds to the cell body of granule cells, oligodendrocytes, but not astrocytes or nerve endings. Identification of granule cells as neuronal ET targets was confirmed by the observation that ET induced intracellular Ca(2+) rises and glutamate release in primary cultures of granule cells. In cultured cerebellar slices, whole cell patch-clamp recordings of synaptic currents in Purkinje cells revealed that ET greatly stimulates both spontaneous excitatory and inhibitory activities. However, pharmacological dissection of these effects indicated that they were only a result of an increased granule cell firing activity and did not involve a direct action of the toxin on glutamatergic nerve terminals or inhibitory interneurons. Patch-clamp recordings of granule cell somata showed that ET causes a decrease in neuronal membrane resistance associated with pore-opening and depolarization of the neuronal membrane, which subsequently lead to the firing of the neuronal network and stimulation of glutamate release. This work demonstrates that a subset of neurons can be directly targeted by ET, suggesting that part of ET-induced neuronal damage observed in neuronal tissue is due to a direct effect of ET on neurons.

  4. Clostridium perfringens Epsilon Toxin Targets Granule Cells in the Mouse Cerebellum and Stimulates Glutamate Release

    PubMed Central

    Lonchamp, Etienne; Dupont, Jean-Luc; Wioland, Laetitia; Courjaret, Raphaël; Mbebi-Liegeois, Corinne; Jover, Emmanuel; Doussau, Frédéric; Popoff, Michel R.; Bossu, Jean-Louis; de Barry, Jean; Poulain, Bernard

    2010-01-01

    Epsilon toxin (ET) produced by C. perfringens types B and D is a highly potent pore-forming toxin. ET-intoxicated animals express severe neurological disorders that are thought to result from the formation of vasogenic brain edemas and indirect neuronal excitotoxicity. The cerebellum is a predilection site for ET damage. ET has been proposed to bind to glial cells such as astrocytes and oligodendrocytes. However, the possibility that ET binds and attacks the neurons remains an open question. Using specific anti-ET mouse polyclonal antibodies and mouse brain slices preincubated with ET, we found that several brain structures were labeled, the cerebellum being a prominent one. In cerebellar slices, we analyzed the co-staining of ET with specific cell markers, and found that ET binds to the cell body of granule cells, oligodendrocytes, but not astrocytes or nerve endings. Identification of granule cells as neuronal ET targets was confirmed by the observation that ET induced intracellular Ca2+ rises and glutamate release in primary cultures of granule cells. In cultured cerebellar slices, whole cell patch-clamp recordings of synaptic currents in Purkinje cells revealed that ET greatly stimulates both spontaneous excitatory and inhibitory activities. However, pharmacological dissection of these effects indicated that they were only a result of an increased granule cell firing activity and did not involve a direct action of the toxin on glutamatergic nerve terminals or inhibitory interneurons. Patch-clamp recordings of granule cell somata showed that ET causes a decrease in neuronal membrane resistance associated with pore-opening and depolarization of the neuronal membrane, which subsequently lead to the firing of the neuronal network and stimulation of glutamate release. This work demonstrates that a subset of neurons can be directly targeted by ET, suggesting that part of ET-induced neuronal damage observed in neuronal tissue is due to a direct effect of ET on neurons. PMID:20941361

  5. Propidium iodide staining: a new application in fluorescence microscopy for analysis of cytoarchitecture in adult and developing rodent brain.

    PubMed

    Hezel, Marcus; Ebrahimi, Fahim; Koch, Marco; Dehghani, Faramarz

    2012-10-01

    Immunohistochemical visualization of antigens in specimen has evolved to an indispensable technique in biomedical research for investigations of cell morphology and pathology both in bright field and fluorescence microscopy. While there are couple of staining methods that reveal entire cytoarchitecture in bright field microscopy such as Nissl or hemalaun-eosin, there are still limitations in visualizations of cytoarchitecture in fluorescence microscopy. The present study reports a simple staining method that provides the required illustration of cell allocations and cellular composition in fluorescence microscopy in adult and in developing rodent central nervous system using the fluorophore propidium iodide (PI, 5μg/mL). PI is a well-accepted marker for degenerating cells when applied prior to fixation (pre-fixation PI staining). Here, PI was added to the sections after the fixation (post-fixation PI staining). This revised labeling procedure led to similar cytoarchitectural staining patterns in fluorescence microscopy as observed with hemalaun in bright field microscopy. This finding was proven in organotypic hippocampal slice cultures (OHSC) and brain sections obtained from different postnatal developmental stages. Excitotoxically lesioned OHSC subjected to pre-fixation PI staining merely showed brightly labeled condensed nuclei of degenerating neurons. In contrast, post-fixation PI staining additionally revealed extensive labeling of neuronal cell bodies and glial cells within the OHSC, thus allowing visualization of stratification of neuronal layers and cell morphology. Furthermore, post-fixation PI staining was combined with NeuN, calbindin, calretinin, glial fibrillary acidic protein or Griffonia simplicifolia isolectin B4 (IB(4)) in post natal (p1 and p9) and adult rats. In early post-natal brain sections almost all mentioned cellular markers led to an incomplete staining of the native cell organization and resulted in an inaccurate estimation of cell morphology when compared to adult brains. In contrast, post-fixation PI staining allowed investigation of the whole cytoarchitecture independent of the developmental stage. Taken together, post-fixation PI staining provides a detailed insight in the morphology of both developing and adult brain tissues in fluorescence microscopy. Copyright © 2012 Elsevier Ltd. All rights reserved.

  6. Not a Stage! A Critical Re-Conception of Young Adolescent Education. Adolescent Cultures, School and Society. Volume 60

    ERIC Educational Resources Information Center

    Vagle, Mark D., Ed.

    2012-01-01

    "Not a Stage!" is written for teachers, students, and scholars interested in the academic, social, and emotional needs of young adolescents. It is unique because it actively resists basing the practice, research, and theory of young adolescent education on developmentalism and the developmental stage of young adolescence. The purpose of…

  7. From big data to deep insight in developmental science

    PubMed Central

    2016-01-01

    The use of the term ‘big data’ has grown substantially over the past several decades and is now widespread. In this review, I ask what makes data ‘big’ and what implications the size, density, or complexity of datasets have for the science of human development. A survey of existing datasets illustrates how existing large, complex, multilevel, and multimeasure data can reveal the complexities of developmental processes. At the same time, significant technical, policy, ethics, transparency, cultural, and conceptual issues associated with the use of big data must be addressed. Most big developmental science data are currently hard to find and cumbersome to access, the field lacks a culture of data sharing, and there is no consensus about who owns or should control research data. But, these barriers are dissolving. Developmental researchers are finding new ways to collect, manage, store, share, and enable others to reuse data. This promises a future in which big data can lead to deeper insights about some of the most profound questions in behavioral science. WIREs Cogn Sci 2016, 7:112–126. doi: 10.1002/wcs.1379 For further resources related to this article, please visit the WIREs website. PMID:26805777

  8. Developmental anatomy of blueberry (Vaccinium corymbosum L. ‘Aurora’) shoot regeneration

    USDA-ARS?s Scientific Manuscript database

    The culture of Vaccinium corymbosum L. ’Aurora’ leaves on regeneration medium results in the regeneration of adventitious shoots. We present anatomical evidence that these new shoot apices are directly regenerated from the cultured blades. Mounds of densely staining cells, which formed from epidermi...

  9. Neighborhood and School Ethnic Structuring and Cultural Adaptations among Mexican-Origin Adolescents

    ERIC Educational Resources Information Center

    White, Rebecca M. B.; Updegraff, Kimberly A.; Umaña-Taylor, Adriana J.; Zeiders, Katharine H.; Perez-Brena, Norma; Burleson, Elizabeth

    2017-01-01

    The ethnic and racial structuring of U.S. neighborhoods may have important implications for developmental competencies during adolescence, including the development of heritage and mainstream cultural orientations. In particular, living in highly concentrated Latino neighborhoods during early adolescence--which channels adolescents into related…

  10. Growth differences and competition between Listeria monocytogenes strains determine their predominance on ham slices and lead to bias during selective enrichment with the ISO protocol.

    PubMed

    Zilelidou, Evangelia; Manthou, Evanthia; Skandamis, Panagiotis

    2016-10-17

    Listeria monocytogenes strains are widespread in the environment where they live well mixed, often resulting in multiple strains contaminating a single food sample. The occurrence of different strains in the same food might trigger strain competition, contributing to uneven growth of strains in food and to bias during selective procedures. We tested the growth of seven L. monocytogenes strains (C5, 6179, ScottA, PL24, PL25, PL26, PL27) on ham slices and on nutrient-rich agar at 10°C, singly and in combinations. Strains were made resistant to different antibiotics for their selective enumeration. In addition, growth of single strains (axenic culture) and competition between strains in xenic cultures of two strains was evaluated in enrichment broth and on selective agar. According to ISO 11290-1:1996/Amd 1:2004 standard protocol for detection of L. monocytogenes, two enrichment steps both followed by streaking on ALOA were performed. Strain cultures were directly added in the enrichment broth or used to inoculate minced beef and sliced hams which were then mixed with enrichment broth. 180-360 colonies were used to determine the relative percentage of each strain recovered on plates per enrichment step. The data showed a significant impact of co-cultivation on the growth of six out of seven strains on ham and a bias towards certain strains during selective enrichment. Competition was manifested by: (i) cessation of growth for the outcompeted strain when the dominant strain reached stationary phase, (ii) reduction of growth rates or (iii) total suppression of growth (both on ham and in enrichment broth or ALOA). Outgrowth of strains by their competitors on ALOA resulted in limited to no recovery, with the outcompeting strain accounting for up to 100% of the total recovered colonies. The observed bias was associated with the enrichment conditions (i.e. food type added to the enrichment broth) and the strain-combination. The outcome of growth competition on food or nonselective agar surface did not necessarily coincide with the results of competition during enrichment. The results show that certain strains present in foods may be missed during classical detection due to strain competition and such likelihood should be taken into consideration when resolving a listeriosis outbreak. Copyright © 2016 Elsevier B.V. All rights reserved.

  11. Culture medium composition affects the gene expression pattern and in vitro development potential of bovine somatic cell nuclear transfer (SCNT) embryos.

    PubMed

    Arias, María E; Ross, Pablo J; Felmer, Ricardo N

    2013-01-01

    Different culture systems have been studied that support development of somatic cell nuclear transfer (SCNT) embryos up to the blastocyst stage. However, the use of sequential and two-step culture systems has been less studied. The objective of the present study was to examine the developmental potential and quality of bovine SCNT embryos cultured in different two-step culture media based on KSOM, SOF and the macromolecules FBS and BSA (K-K/FBS, K-S/BSA and K-K/BSA, respectively). No differences were observed in the cleavage rate for any of the culture systems. However, there was a significant difference (P<0.01) in the rate of blastocyst development, with the K-K/ FBS culture system yielding a higher rate of blastocysts (28%) compared to other treatments (18 and 15%, for K-S/BSA and K-K/BSA, respectively). Although quality of embryos, as assessed by the total number of cells, was not different, the apoptosis index was significantly affected in the sequential culture system (K-S/BSA). Gene expression analysis showed alterations of DNMT1, IGF2, LIF, and PRDX6 genes in embryos cultured in K-S/FBS and of SOD2 in embryos cultured in K-K/BSA. In conclusion, we demonstrated that culture medium may affect not only the developmental potential of SCNT embryos but also, more importantly, the gene expression pattern and apoptotic index, presenting the possibility to manipulate the culture medium composition to modulate global gene expression and improve the overall efficiency of this technique.

  12. Freezing effect on bread appearance evaluated by digital imaging

    NASA Astrophysics Data System (ADS)

    Zayas, Inna Y.

    1999-01-01

    In marketing channels, bread is sometimes delivered in a frozen sate for distribution. Changes occur in physical dimensions, crumb grain and appearance of slices. Ten loaves, twelve bread slices per loaf were scanned for digital image analysis and then frozen in a commercial refrigerator. The bread slices were stored for four weeks scanned again, permitted to thaw and scanned a third time. Image features were extracted, to determine shape, size and image texture of the slices. Different thresholds of grey levels were set to detect changes that occurred in crumb, images were binarized at these settings. The number of pixels falling into these gray level settings were determined for each slice. Image texture features of subimages of each slice were calculated to quantify slice crumb grain. The image features of the slice size showed shrinking of bread slices, as a results of freezing and storage, although shape of slices did not change markedly. Visible crumb texture changes occurred and these changes were depicted by changes in image texture features. Image texture features showed that slice crumb changed differently at the center of a slice compared to a peripheral area close to the crust. Image texture and slice features were sufficient for discrimination of slices before and after freezing and after thawing.

  13. Theory of mind and executive function in Chinese preschool children.

    PubMed

    Duh, Shinchieh; Paik, Jae H; Miller, Patricia H; Gluck, Stephanie C; Li, Hui; Himelfarb, Igor

    2016-04-01

    Cross-cultural research on children's theory of mind (ToM) understanding has raised questions about its developmental sequence and relationship with executive function (EF). The current study examined how ToM develops (using the tasks from Wellman & Liu, 2004) in relation to 2 EF skills (conflict inhibition, working memory) in 997 Chinese preschoolers (ages 3, 4, 5) in Chengdu, China. Compared with prior research with other Chinese and non-Chinese children, some general patterns in development were replicated in this sample. However, the children showed culture-specific reversals in the developmental sequence of ToM. For example, Chengdu children performed differently on the 2 false-belief tasks that were thought to be equivalent. Furthermore, conflict inhibition as well as working memory uniquely predicted ToM performance. We discuss the issues of ToM development as they relate to test items and cross-cultural--and subcultural--differences. (c) 2016 APA, all rights reserved).

  14. Understanding Chinese Immigrant and European American Mothers' Expressions of Warmth

    ERIC Educational Resources Information Center

    Cheah, Charissa S. L.; Li, Jin; Zhou, Nan; Yamamoto, Yoko; Leung, Christy Y. Y.

    2015-01-01

    Maternal warmth, the quality of the affectional bond between mothers and their children, has been found to be consistently associated with children's positive developmental outcomes in Western cultures. However, researchers debate the potential differences in the cultural meanings of maternal warmth, particularly between Chinese and European…

  15. Psychometric Examination of a Spanish Translation of a Developmental Screening Instrument

    ERIC Educational Resources Information Center

    Pomés, Maria; Squires, Jane; Yovanoff, Paul

    2016-01-01

    Immigrant populations are growing and permanently changing the demographic profile of the United States. Diverse cultural and linguistic backgrounds are imposing demands and challenges upon agencies serving young children and families. Culturally sensitive assessments are not always available for these populations, and psychometric properties of…

  16. Target Article with Commentaries: Developmental Niche Construction

    ERIC Educational Resources Information Center

    Flynn, Emma G.; Laland, Kevin N.; Kendal, Rachel L.; Kendal, Jeremy R.

    2013-01-01

    Niche construction is the modification of components of the environment through an organism's activities. Humans modify their environments mainly through ontogenetic and cultural processes, and it is this reliance on learning, plasticity and culture that lends human niche construction a special potency. In this paper we aim to facilitate…

  17. When Does Social Learning Become Cultural Learning?

    ERIC Educational Resources Information Center

    Heyes, Cecilia

    2017-01-01

    Developmental research on selective social learning, or "social learning strategies", is currently a rich source of information about when children copy behaviour, and who they prefer to copy. It also has the potential to tell us when and how human social learning becomes cultural learning; i.e. mediated by psychological mechanisms that…

  18. Using Naming Practices in the Developmental English Classroom

    ERIC Educational Resources Information Center

    Behrens, Susan

    2010-01-01

    Linguistic behavior often reveals cultural practices. In fact, sociolinguistics is dedicated to researching how language use intersects with cultural and social identity (e.g., Coupland. and Jaworski, 2009). One aspect of language use that sociolinguistics focuses on is naming practices. The study of such practices is called onomastics. While…

  19. The Development of Generosity and Moral Cognition across Five Cultures

    ERIC Educational Resources Information Center

    Cowell, Jason M.; Lee, Kang; Malcolm-Smith, Susan; Selcuk, Bilge; Zhou, Xinyue; Decety, Jean

    2017-01-01

    Morality is an evolved aspect of human nature, yet is heavily influenced by cultural environment. This developmental study adopted an integrative approach by combining measures of socioeconomic status (SES), executive function, affective sharing, empathic concern, theory of mind, and moral judgment in predicting sharing behavior in children (N =…

  20. Comparative sensitivity of human and rat neural cultures to chemical-induced inhibition of neurite outgrowth

    EPA Science Inventory

    There is a need for rapid, efficient and cost effective alternatives to traditional in vivo developmental neurotoxicity testing. In vitro cell culture models can recapitulate many of the key cellular processes of nervous system development, including neurite outgrowth, and may be...

  1. ASSESSMENT OF CHEMICAL EFFECTS ON NEURITE OUTGROWTH, NEURONAL POLARIZATION AND SYNAPTOGENESIS IN RAT CORTICAL NEURONS USING HIGH CONTENT IMAGE ANALYSIS

    EPA Science Inventory

    There is a need for efficient, cost-effective methods for screening and prioritization of potential developmental neurotoxicants. One approach uses in vitro cell culture models that can recapitulate the critical processes of nervous system development. In vitro, primary cultures ...

  2. Childhood Gender Identity...Disorder? Developmental, Cultural, and Diagnostic Concerns

    ERIC Educational Resources Information Center

    Dragowski, Eliza A.; Scharron-del Rio, Maria R.; Sandigorsky, Amy L.

    2011-01-01

    Childhood gender identity development is reviewed in the context of biological, environmental, cultural, and diagnostic factors. With the upcoming 5th revision of the "Diagnostic and Statistical Manual of Mental Disorders," the authors offer a critical consideration of childhood gender identity disorder, along with proposed diagnostic changes.…

  3. Musical Acculturation through Primary School Activities during Japanese Colonial Rule of Korea (1910-1945)

    ERIC Educational Resources Information Center

    Kim, Jeong Ha

    2014-01-01

    Global colonialism and continuing post-colonial influences caused widespread cultural change at the interface of different cultures. Musical acculturation can be observed in most colonised countries. Some pro-colonialists apologetically allege that through colonisation the colonised territories would receive developmental aid and economical…

  4. Rapid Single-Cell Electroporation for Labeling Organotypic Cultures

    DTIC Science & Technology

    2010-06-01

    carry out SCE on the tissue slice under visual control from the user. When the user wishes to transfect a new compound , the entire micromanipulator...a new compound by a perpendicular long-travel linear stage (H). Each sub-system is discussed below in detail including development and choice of...stage where a cleaning bath is moved in. The tip is inserted into the bath and cleaned/rinsed (E) before sampling a new compound and repeating (A). Af D

  5. Combating School Bullying through Developmental Guidance for Positive Youth Development and Promoting Harmonious School Culture

    PubMed Central

    Hui, Eadaoin K. P.; Tsang, Sandra K. M.; Law, Bella C. M.

    2011-01-01

    Bullying and violence, which can bring detrimental effects, are situations which young people have to face in their process of development. Though school bullying has been a spreading and explicit problem in Hong Kong schools, most of the programs or guidelines dealing with the problem lack citywide, recognized initiatives and the effectiveness of these programs is unknown due to the lack of evaluation. The present paper discusses preventing school bullying from a developmental guidance perspective, using the positive youth development paradigm and promoting the values of harmony and forgiveness at the whole-school level to cultivate a harmonious school culture as a way of combating school bullying. PMID:22194662

  6. Combating school bullying through developmental guidance for positive youth development and promoting harmonious school culture.

    PubMed

    Hui, Eadaoin K P; Tsang, Sandra K M; Law, Bella C M

    2011-01-01

    Bullying and violence, which can bring detrimental effects, are situations which young people have to face in their process of development. Though school bullying has been a spreading and explicit problem in Hong Kong schools, most of the programs or guidelines dealing with the problem lack citywide, recognized initiatives and the effectiveness of these programs is unknown due to the lack of evaluation. The present paper discusses preventing school bullying from a developmental guidance perspective, using the positive youth development paradigm and promoting the values of harmony and forgiveness at the whole-school level to cultivate a harmonious school culture as a way of combating school bullying.

  7. Perceptions of societal developmental hierarchies in Europe and beyond: A Bulgarian Perspective

    PubMed Central

    Melegh, Attila; Thornton, Arland; Philipov, Dimiter; Young-DeMarco, Linda

    2012-01-01

    We examine how ordinary citizens in Bulgaria view the developmental levels of European countries and certain states outside of Europe. Our research is motivated by the understanding that scholars and policy makers have for centuries used developmental hierarchies to characterize countries and that this perception of differential development has shaped interactions among different groups, countries and regions. We expect that views of such developmental hierarchies and models have great potential for influencing demographic and family behavior and political and cultural identities of ordinary people. Using data from a 2009 survey in Bulgaria we document that developmental hierarchies are widely perceived in Bulgaria, but are distributed differentially by age, education, and degree of urbanization. We also consider internal mechanisms underlying this hierarchical understanding of development and how hierarchical understandings may be related to national identities. PMID:23807821

  8. Precision-cut liver slices to investigate responsiveness of deep-sea fish to contaminants at high pressure.

    PubMed

    Lemaire, Benjamin; Debier, Cathy; Calderon, Pedro Buc; Thomé, Jean Pierre; Stegeman, John; Mork, Jarle; Rees, Jean François

    2012-09-18

    While deep-sea fish accumulate high levels of persistent organic pollutants (POPs), the toxicity associated with this contamination remains unknown. Indeed, the recurrent collection of moribund individuals precludes experimental studies to investigate POP effects in this fauna. We show that precision-cut liver slices (PCLS), an in vitro tool commonly used in human and rodent toxicology, can overcome such limitation. This technology was applied to individuals of the deep-sea grenadier Coryphaenoides rupestris directly upon retrieval from 530-m depth in Trondheimsfjord (Norway). PCLS remained viable and functional for 15 h when maintained in an appropriate culture media at 4 °C. This allowed experimental exposure of liver slices to the model POP 3-methylcholanthrene (3-MC; 25 μM) at levels of hydrostatic pressure mimicking shallow (0.1 megapascal or MPa) and deep-sea (5-15 MPa; representative of 500-1500 m depth) environments. As in shallow water fish, 3-MC induced the transcription of the detoxification enzyme cytochrome P4501A (CYP1A; a biomarker of exposure to POPs). This induction was diminished at elevated pressure, suggesting a limited responsiveness of C. rupestris toward POPs in its native environment. This very first in vitro toxicological investigation on a deep-sea fish opens the route for understanding pollutants effects in this highly exposed fauna.

  9. Contamination of knives and graters by bacterial foodborne pathogens during slicing and grating of produce.

    PubMed

    Erickson, Marilyn C; Liao, Jean; Cannon, Jennifer L; Ortega, Ynes R

    2015-12-01

    Poor hygiene and improper food preparation practices in consumers' homes have previously been demonstrated as contributing to foodborne diseases. To address potential cross-contamination by kitchen utensils in the home, a series of studies was conducted to determine the extent to which the use of a knife or grater on fresh produce would lead to the utensil's contamination with Escherichia coli O157:H7 or Salmonella enterica. When shredding inoculated carrots (ca. 5.3 log CFU/carrot), all graters became contaminated and the number of E. coli O157:H7 present on the utensil was significantly greater than Salmonella (p < 0.05). Contamination of knives after slicing inoculated produce (4.9-5.4 log CFU/produce item) could only be detected by enrichment culture. After slicing tomatoes, honeydew melons, strawberries, cucumbers, and cantaloupes, the average prevalence of knife contamination by the two pathogens was 43%, 17%, 15%, 7%, and 3%, respectively. No significant increase in the incidence or level of contamination occurred on the utensils when residues were present (p > 0.05); however, subsequent contamination of 7 produce items processed with the contaminated utensils did occur. These results highlight the necessity of proper sanitization of these utensils when used in preparation of raw produce. Copyright © 2015 Elsevier Ltd. All rights reserved.

  10. Studies on the neuroprotective action of kynurenine mono-oxygenase inhibitors in post-ischemic brain damage.

    PubMed

    Moroni, Flavio; Carpenedo, Raffaella; Cozzi, Andrea; Meli, Elena; Chiarugi, Alberto; Pellegrini-Giampietro, Domenico E

    2003-01-01

    Kynurenine 3-mono-oxygenase (KMO) inhibitors facilitate kynurenic acid (KYNA) neosynthesis and reduce the formation of 3OH-kynurenine (3-HK) and quinolinic acid (QUIN). They also attenuate post-ischemic brain damage and decrease glutamate (Glu) content in brain extracellular spaces. To investigate KMO mechanism(s) of neuroprotection, we performed experiments in gerbils subjected to bilateral carotid occlusion and in organotypic rat hippocampal slice cultures exposed to oxygen and glucose deprivation (OGD). In gerbils, direct application of KYNA (100 nM, through reverse microdialysis in the hippocampus) completely prevented the increase in Glu output induced by transient (5 min) occlusion of the carotids. In rat hippocampal slices exposed for 30 min to OGD, KMO inhibitors (m-nitrobenzoyl)-alanine (mNBA, 30-100 microM) or 3,4-dimethoxy-[-N-4-(nitrophenyl)thiazol-2yl]-benzenesulfonamide (Ro 61-8048, 1-10 microM) reduced post-ischemic neuronal death and increased KYNA concentrations in the incubation medium. KYNA may antagonize glycineb or alpha7 nicotinic acetylcholine receptors but the concentrations in the incubation medium never reached values that could efficiently antagonize receptor function. On the contrary, 3-HK (1-10 microM) added to slices exposed to OGD in the presence of KMO inhibitors completely prevented the neuroprotective effects of the inhibitors. Our findings suggest that KMO inhibitors reduce OGD-induced pyramidal cell death by decreasing 3-HK (and possibly QUIN) synthesis.

  11. Live dynamic imaging and analysis of developmental cardiac defects in mouse models with optical coherence tomography

    NASA Astrophysics Data System (ADS)

    Lopez, Andrew L.; Wang, Shang; Garcia, Monica; Valladolid, Christian; Larin, Kirill V.; Larina, Irina V.

    2015-03-01

    Understanding mouse embryonic development is an invaluable resource for our interpretation of normal human embryology and congenital defects. Our research focuses on developing methods for live imaging and dynamic characterization of early embryonic development in mouse models of human diseases. Using multidisciplinary methods: optical coherence tomography (OCT), live mouse embryo manipulations and static embryo culture, molecular biology, advanced image processing and computational modeling we aim to understand developmental processes. We have developed an OCT based approach to image live early mouse embryos (E8.5 - E9.5) cultured on an imaging stage and visualize developmental events with a spatial resolution of a few micrometers (less than the size of an individual cell) and a frame rate of up to hundreds of frames per second and reconstruct cardiodynamics in 4D (3D+time). We are now using these methods to study how specific embryonic lethal mutations affect cardiac morphology and function during early development.

  12. Alignment of governance and senior executive perceptions of culture.

    PubMed

    Looi, Evelyn Suk Yi; Greatbanks, Richard; Everett, André M

    2016-09-19

    Purpose The purpose of this paper is to examine the alignment of perceived organizational culture between Health Board chairs and Board members with that of their respective senior executive teams. It compares the degree of alignment between these two groups, and analyses them against District Health Board (DHB) performance using the New Zealand Ministry of Health's national measure "shorter stays in Emergency Departments." Design/methodology/approach Primary survey data were collected across eight DHBs using a modified version of the Organizational Culture Assessment Instrument and utilizes a sample of both higher and lower performing DHBs as defined by the "shorter stays" measure. Findings Many health organizations cite Ovseiko and Buchan's (2012) preferred culture as an ideal model. However, this study's findings indicate that most DHBs scored higher than the preferred score for "Hierarchical" and "Rational" cultures, and lower for "Clan" and "Developmental" cultures, and therefore calls into question the validity of this organizational profile as the "preferred" cultural state. Research limitations/implications This research considers perceived organizational culture from the perspective of the Board members and their respective senior executive teams. It uses a relatively small sample size and excludes potential interactions of national culture. Practical implications The findings indicate that healthcare organizations should de-emphasize the dominant "Hierarchical" and "Rational" cultures, and promote "Clan" and "Developmental" cultures within their organizations as a means of potentially improving healthcare performance. Originality/value Organizational culture has been highlighted as a major component of performance within healthcare organizations, yet very few studies examine how organizational culture is perceived by governance and executive groups. This study empirically counters prevailing knowledge regarding the most appropriate organizational cultures for healthcare organizations.

  13. Neuroprotection by JM-20 against oxygen-glucose deprivation in rat hippocampal slices: Involvement of the Akt/GSK-3β pathway.

    PubMed

    Ramírez-Sánchez, Jeney; Simões Pires, Elisa Nicoloso; Nuñez-Figueredo, Yanier; Pardo-Andreu, Gilberto L; Fonseca-Fonseca, Luis Arturo; Ruiz-Reyes, Alberto; Ochoa-Rodríguez, Estael; Verdecia-Reyes, Yamila; Delgado-Hernández, René; Souza, Diogo O; Salbego, Christianne

    2015-11-01

    Cerebral ischemia is the third most common cause of death and a major cause of disability worldwide. Beyond a shortage of essential metabolites, ischemia triggers many interconnected pathophysiological events, including excitotoxicity, oxidative stress, inflammation and apoptosis. Here, we investigated the neuroprotective mechanisms of JM-20, a novel synthetic molecule, focusing on the phosphoinositide-3-kinase (PI3K)/Akt survival pathway and glial cell response as potential targets of JM-20. For this purpose, we used organotypic hippocampal slice cultures exposed to oxygen-glucose deprivation (OGD) to achieve ischemic/reperfusion damage in vitro. Treatment with JM-20 at 0.1 and 10 μM reduced PI incorporation (indicative of cell death) after OGD. OGD decreased the phosphorylation of Akt (pro-survival) and GSK 3β (pro-apoptotic), resulting in respective inhibition and activation of these proteins. Treatment with JM20 prevented the reduced phosphorylation of these proteins after OGD, representing a shift from pro-apoptotic to pro-survival signaling. The OGD-induced activation of caspase-3 was also attenuated by JM-20 treatment at 10 μM. Moreover, in cultures treated with JM-20 and exposed to OGD conditioning, we observed a decrease in activated microglia, as well as a decrease in interleukin (IL)-1β, IL-6 and tumor necrosis factor (TNF)-α release into the culture medium, while the level of the anti-inflammatory IL-10 increased. GFAP immunostaining and IB4 labeling showed that JM-20 treatment significantly augmented GFAP immunoreactivity after OGD, when compared with cultures exposed to OGD only, suggesting the activation of astroglial cells. Our results confirm that JM-20 has a strong neuroprotective effect against ischemic injury and suggest that the mechanisms involved in this effect may include the modulation of reactive astrogliosis, as well as neuroinflammation and the anti-apoptotic cell signaling pathway. Copyright © 2015 Elsevier Ltd. All rights reserved.

  14. Induction of Endothelial Phenotype From Wharton's Jelly-Derived MSCs and Comparison of Their Vasoprotective and Neuroprotective Potential With Primary WJ-MSCs in CA1 Hippocampal Region Ex Vivo.

    PubMed

    Obtulowicz, Patrycja; Lech, Wioletta; Strojek, Lukasz; Sarnowska, Anna; Domanska-Janik, Krystyna

    2016-01-01

    Ischemic stroke results in violent impairment of tissue homeostasis leading to severe perturbation within the neurovascular unit (NVU) during the recovery period. The aim of this study was to assess the potential of mesenchymal stem cells (MSCs) originating from Wharton's jelly (WJ) to differentiate into functionally competent cells of endothelial lineage (WJ-EPCs). The protective effect(s) of either primary WJ-MSCs or induced WJ-EPCs was investigated and compared after oxygen-glucose deprivation (OGD) of hippocampal organotypic slices (OHC) in the indirect coculture model. WJ-MSCs, primed in EGM-2 (Lonza commercial medium) under 5% O2, acquired cobblestone endothelial-like morphology, formed capillary-like structures and actively took up DiI-Ac-LDL. Both cell types (WJ-MSCs and WJ-EPCs) were positive for CD73, CD90, CD105, VEGFR-2, and VEGF, but only endothelial-like culture expressed vWF and PECAM-1 markers at significant levels. In the presence of either WJ-MSCs or WJ-EPCs in the compartment below OGD-injured slices, cell death and vascular atrophy in the hypoxia-sensitive CA1 region were substantially decreased. This suggests that a paracrine mechanism may mediate WJ-MSC- and WJ-EPC-dependent protection. Thus, finally, we estimated secretion of the neuro/angio/immunomodulatory molecules IL-6, TGF-β1, and VEGF by these cell cultures. We have found that release of TGF-β1 and IL-6 was TLR ligand [LPS and Poly(I:C)] concentration dependent and stronger in WJ-EPC than WJ-MSC cultures. Simultaneously, the uneven pattern of TLR receptors and modulatory cytokine gene expression was confirmed also on qRT-PCR level, but no significant differences were noticed between WJ-EPC and primary WJ-MSC cultures.

  15. Partners | Office of Cancer Clinical Proteomics Research

    Cancer.gov

    Developmental Studies Hybridoma Bank at the University of Iowa NCI’s OCCPR works closely with The University of Iowa's Developmental Studies Hybridoma Bank (DSHB) that distributes all hybridomas and monoclonal antibodies from NCI's Clinical Proteomic Technologies for Cancer initiative (CPTC). DSHB supplies researchers with monoclonal antibodies, which may be ordered as tissue culture supernatants, ascites, or concentrate; selected hybridomas are also available as frozen or growing cells.

  16. Cross-Cultural Issues in Parent-Professional Interactions: A Qualitative Study of Perceptions of Asian American Mothers of Children with Developmental Disabilities

    ERIC Educational Resources Information Center

    Jegatheesan, Brinda

    2009-01-01

    This study investigated the perspectives of 23 first-generation Asian American mothers of children with developmental disabilities. The intent was to explore the working relationships between the mothers and professionals in health care and special education in the United States. The participants in this study were from China, Taiwan, Vietnam,…

  17. Towards New Cultures of Learning: Personal Learning Environments as a Developmental Perspective for Improving Higher Education Language Courses

    ERIC Educational Resources Information Center

    Laakkonen, Ilona; Taalas, Peppi

    2015-01-01

    This article provides readers with an understanding of the concept of the personal learning environment (PLE). It suggests that PLEs can be used in two complementary ways: as a developmental lens for integrating ICT and creating new pedagogical practices and digital literacies for academic language learning, and as a context in which learners can…

  18. Living without Restraint: One Parent's Reflections and Recommendations for Supporting At-Risk Individuals with Developmental Disabilities

    ERIC Educational Resources Information Center

    Beaudoin, Wilfred; Moore, Adam

    2018-01-01

    In the past several years, there has been an important movement to reduce the utilization of restraint for individuals with developmental disabilities. Legislatures, local and national, are taking on the task of shaping the way that our culture supports people who, up until now, have been often treated in a punitive manner rather than truly…

  19. Current economic and sensitivity analysis for ID slicing of 4 inch and 6 inch diameter silicon ingots for photovoltaic applications

    NASA Technical Reports Server (NTRS)

    Roberts, E. G.; Johnson, C. M.

    1982-01-01

    The economics and sensitivities of slicing large diameter silicon ingots for photovoltaic applications were examined. Current economics and slicing add on cost sensitivities are calculated using variable parameters for blade life, slicing yield, and slice cutting speed. It is indicated that cutting speed has the biggest impact on slicing add on cost, followed by slicing yield, and by blade life as the blade life increases.

  20. Targeting of astrocytic glucose metabolism by beta-hydroxybutyrate.

    PubMed

    Valdebenito, Rocío; Ruminot, Iván; Garrido-Gerter, Pamela; Fernández-Moncada, Ignacio; Forero-Quintero, Linda; Alegría, Karin; Becker, Holger M; Deitmer, Joachim W; Barros, L Felipe

    2016-10-01

    The effectiveness of ketogenic diets and intermittent fasting against neurological disorders has brought interest to the effects of ketone bodies on brain cells. These compounds are known to modify the metabolism of neurons, but little is known about their effect on astrocytes, cells that control the supply of glucose to neurons and also modulate neuronal excitability through the glycolytic production of lactate. Here we have used genetically-encoded Förster Resonance Energy Transfer nanosensors for glucose, pyruvate and ATP to characterize astrocytic energy metabolism at cellular resolution. Our results show that the ketone body beta-hydroxybutyrate strongly inhibited astrocytic glucose consumption in mouse astrocytes in mixed cultures, in organotypic hippocampal slices and in acute hippocampal slices prepared from ketotic mice, while blunting the stimulation of glycolysis by physiological and pathophysiological stimuli. The inhibition of glycolysis was paralleled by an increased ability of astrocytic mitochondria to metabolize pyruvate. These results support the emerging notion that astrocytes contribute to the neuroprotective effect of ketone bodies. © The Author(s) 2015.

  1. Family support and intergenerational caregiving: report from the State of the Science in Aging with Developmental Disabilities Conference.

    PubMed

    Heller, Tamar; Factor, Alan

    2008-07-01

    This article summarizes the proceedings of the Family Support and Intergenerational Caregiving work group that was part of the "State of the Science in Aging with Developmental Disabilities: Charting Lifespan Trajectories and Supportive Environments for Healthy Living" symposium. The aim was to provide a research and policy agenda that would result in a better social, health, and economic well being for adults with intellectual and developmental disabilities and family caregivers. Key issues addressed were (1) demographic challenges; (2) long-term outcomes, poverty and cultural minority status; (3) interdependent roles of family members; and (4) family support policies and practices. The group identified the following major gaps in knowledge that need to be included in a future research agenda: data on the demography of families, including information on cultural minorities and families living in poverty; research on the interdependent roles of various family members in providing support to each other, including siblings, grandparents, and the person with disabilities; and development and assessment of models for providing supports to families and that bridge the aging and disability networks. The outcomes of this recommended research agenda will result in progressive policies and culturally sensitive supports that will improve the health, social, and economic well-being of adults with I/DD and their families.

  2. Dissociation of Per1 and Bmal1 circadian rhythms in the suprachiasmatic nucleus in parallel with behavioral outputs

    PubMed Central

    Ono, Daisuke; Honma, Sato; Nakajima, Yoshihiro; Kuroda, Shigeru; Enoki, Ryosuke; Honma, Ken-ichi

    2017-01-01

    The temporal order of physiology and behavior in mammals is primarily regulated by the circadian pacemaker located in the hypothalamic suprachiasmatic nucleus (SCN). Taking advantage of bioluminescence reporters, we monitored the circadian rhythms of the expression of clock genes Per1 and Bmal1 in the SCN of freely moving mice and found that the rate of phase shifts induced by a single light pulse was different in the two rhythms. The Per1-luc rhythm was phase-delayed instantaneously by the light presented at the subjective evening in parallel with the activity onset of behavioral rhythm, whereas the Bmal1-ELuc rhythm was phase-delayed gradually, similar to the activity offset. The dissociation was confirmed in cultured SCN slices of mice carrying both Per1-luc and Bmal1-ELuc reporters. The two rhythms in a single SCN slice showed significantly different periods in a long-term (3 wk) culture and were internally desynchronized. Regional specificity in the SCN was not detected for the period of Per1-luc and Bmal1-ELuc rhythms. Furthermore, neither is synchronized with circadian intracellular Ca2+ rhythms monitored by a calcium indicator, GCaMP6s, or with firing rhythms monitored on a multielectrode array dish, although the coupling between the circadian firing and Ca2+ rhythms persisted during culture. These findings indicate that the expressions of two key clock genes, Per1 and Bmal1, in the SCN are regulated in such a way that they may adopt different phases and free-running periods relative to each other and are respectively associated with the expression of activity onset and offset. PMID:28416676

  3. Heat shock protein 70 in the rat nasal cavity: localisation and response to hyperthermia.

    PubMed

    Simpson, Sharon A; Alexander, David J; Reed, Celia J

    2004-06-01

    Heat shock proteins (HSPs) are a group of proteins that are rapidly induced in response to physiological stress, including hyperthermia and exposure to toxicants. Thus they may provide a useful index of toxicity in in vitro systems for screening for toxicity. We have recently developed a rat nasal explant system for investigating upper respiratory tract toxicity, and the aims of this study were to localise HSP70 within the rat nasal cavity and to characterise its response to hyperthermia. Constitutively, HSP70 was found to be predominantly localised to the sustentacular cells, basal cells and Bowman's glands of the olfactory epithelium (OE), with the most intense immunohistochemical staining at levels 3 and 4 of the posterior of the rat nasal cavity. Ethmoturbinates (ETs) and liver slices were exposed to heat shock (37 degrees and 43 degrees C, respectively) for 45 min and then returned to normal culture temperatures (31 degrees and 37 degrees C, respectively) for 24 h. In ETs, HSP72 was maximally induced 4-fold at 4 h after heat shock, and levels then returned to those of control tissue. ATP concentrations were markedly decreased up to 4 h after heat shock and then returned to control levels. In contrast, HSP72 levels in liver slices increased and ATP levels decreased steadily throughout the 24 h culture period. ETs were also able to withstand a 45-min heat shock at 43 degrees C, that is 12 degrees C above normal culture temperature. Incubation of ETs with cycloheximide prior to heat shock reduced the ability of the OE to recover from heat shock at 37 degrees C. Thus the OE of the rat nasal cavity expresses HSP72, and this protein appears to play an important role in the ability of the tissue to withstand hyperthermia.

  4. Assessment of the Effects of Endocrine Disrupting Compounds on the Development of Vertebrate Neural Network Function Using Multi-electrode Arrays.

    PubMed

    Sanchez, Karla R; Mersha, Mahlet D; Dhillon, Harbinder S; Temburni, Murali K

    2018-04-26

    Bis-phenols, such as bis-phenol A (BPA) and bis-phenol-S (BPS), are polymerizing agents widely used in the production of plastics and numerous everyday products. They are classified as endocrine disrupting compounds (EDC) with estradiol-like properties. Long-term exposure to EDCs, even at low doses, has been linked with various health defects including cancer, behavioral disorders, and infertility, with greater vulnerability during early developmental periods. To study the effects of BPA on the development of neuronal function, we used an in vitro neuronal network derived from the early chick embryonic brain as a model. We found that exposure to BPA affected the development of network activity, specifically spiking activity and synchronization. A change in network activity is the crucial link between the molecular target of a drug or compound and its effect on behavioral outcome. Multi-electrode arrays are increasingly becoming useful tools to study the effects of drugs on network activity in vitro. There are several systems available in the market and, although there are variations in the number of electrodes, the type and quality of the electrode array and the analysis software, the basic underlying principles, and the data obtained is the same across the different systems. Although currently limited to analysis of two-dimensional in vitro cultures, these MEA systems are being improved to enable in vivo network activity in brain slices. Here, we provide a detailed protocol for embryonic exposure and recording neuronal network activity and synchrony, along with representative results.

  5. Visualization of local Ca2+ dynamics with genetically encoded bioluminescent reporters.

    PubMed

    Rogers, Kelly L; Stinnakre, Jacques; Agulhon, Cendra; Jublot, Delphine; Shorte, Spencer L; Kremer, Eric J; Brûlet, Philippe

    2005-02-01

    Measurements of local Ca2+ signalling at different developmental stages and/or in specific cell types is important for understanding aspects of brain functioning. The use of light excitation in fluorescence imaging can cause phototoxicity, photobleaching and auto-fluorescence. In contrast, bioluminescence does not require the input of radiative energy and can therefore be measured over long periods, with very high temporal resolution. Aequorin is a genetically encoded Ca(2+)-sensitive bioluminescent protein, however, its low quantum yield prevents dynamic measurements of Ca2+ responses in single cells. To overcome this limitation, we recently reported the bi-functional Ca2+ reporter gene, GFP-aequorin (GA), which was developed specifically to improve the light output and stability of aequorin chimeras [V. Baubet, et al., (2000) PNAS, 97, 7260-7265]. In the current study, we have genetically targeted GA to different microdomains important in synaptic transmission, including to the mitochondrial matrix, endoplasmic reticulum, synaptic vesicles and to the postsynaptic density. We demonstrate that these reporters enable 'real-time' measurements of subcellular Ca2+ changes in single mammalian neurons using bioluminescence. The high signal-to-noise ratio of these reporters is also important in that it affords the visualization of Ca2+ dynamics in cell-cell communication in neuronal cultures and tissue slices. Further, we demonstrate the utility of this approach in ex-vivo preparations of mammalian retina, a paradigm in which external light input should be controlled. This represents a novel molecular imaging approach for non-invasive monitoring of local Ca2+ dynamics and cellular communication in tissue or whole animal studies.

  6. Argonaute identity defines the length of mature mammalian microRNAs.

    PubMed

    Juvvuna, Prasanna Kumar; Khandelia, Piyush; Lee, Li Ming; Makeyev, Eugene V

    2012-08-01

    MicroRNAs (miRNAs) are 19- to 25-nt-long non-coding RNAs that regulate gene expression by base-pairing with target mRNAs and reducing their stability or translational efficiency. Mammalian miRNAs function in association with four closely related Argonaute proteins, AGO1-4. All four proteins contain the PAZ and the MID domains interacting with the miRNA 3' and 5' termini, respectively, as well as the PIWI domain comprising an mRNA 'slicing' activity in the case of AGO2 but not AGO1, AGO3 and AGO4. However, the slicing mode of the miRNA-programmed AGO2 is rarely realized in vivo and the four Argonautes are thought to play largely overlapping roles in the mammalian miRNA pathway. Here, we show that the average length of many miRNAs is diminished during nervous system development as a result of progressive shortening of the miRNA 3' ends. We link this modification with an increase in the fractional abundance of Ago2 in the adult brain and identify a specific structural motif within the PAZ domain that enables efficient trimming of miRNAs associated with this but not the other three Argonautes. Taken together, our data suggest that mammalian Argonautes may define the length and possibly biological activity of mature mammalian miRNAs in a developmentally controlled manner.

  7. Child and Ancient Man: How to Define Their Commonalities and Differences.

    PubMed

    Oesterdiekhoff, Georg W

    2016-09-01

    Developmental psychology is not only a psychology of development from childhood to old age but a psychology of human development in world history. Eighty years of cross-cultural empirical research findings indicate that the adolescent stage of formal operations evolved late in history and is not a universal development of adult humans across cultures and history. Correspondingly, preoperational or concrete operational stages describe adult psychological stages in past or premodern cultures, as Jean Piaget and some of his followers have mentioned. Developmental psychology is likewise a historical or anthropological psychology capable of describing humans in premodern cultures. The article develops a general anthropological or psychological theory answering the many questions that arise from the correspondences between (modern) children and ancient adults. On this psychological basis, the new structural genetic theory program is capable of explaining, better than previous approaches, the history of humankind from prehistory through ancient to modern societies, the history of economy, society, culture, religion, philosophy, sciences, morals, and everyday life. The accomplishment of this task was once demanded of some classical founders of psychology, sociology, history, and ethnology but was largely avoided by the postwar generations of authors for political and ideological reasons.

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

    PubMed

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

    2013-05-01

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

  9. Insights in spatio-temporal characterization of human fetal neural stem cells.

    PubMed

    Martín-Ibáñez, Raquel; Guardia, Inés; Pardo, Mónica; Herranz, Cristina; Zietlow, Rike; Vinh, Ngoc-Nga; Rosser, Anne; Canals, Josep M

    2017-05-01

    Primary human fetal cells have been used in clinical trials of cell replacement therapy for the treatment of neurodegenerative disorders such as Huntington's disease (HD). However, human fetal primary cells are scarce and difficult to work with and so a renewable source of cells is sought. Human fetal neural stem cells (hfNSCs) can be generated from human fetal tissue, but little is known about the differences between hfNSCs obtained from different developmental stages and brain areas. In the present work we characterized hfNSCs, grown as neurospheres, obtained from three developmental stages: 4-5, 6-7 and 8-9weeks post conception (wpc) and four brain areas: forebrain, cortex, whole ganglionic eminence (WGE) and cerebellum. We observed that, as fetal brain development proceeds, the number of neural precursors is diminished and post-mitotic cells are increased. In turn, primary cells obtained from older embryos are more sensitive to the dissociation process, their viability is diminished and they present lower proliferation ratios compared to younger embryos. However, independently of the developmental stage of derivation proliferation ratios were very low in all cases. Improvements in the expansion rates were achieved by mechanical, instead of enzymatic, dissociation of neurospheres but not by changes in the seeding densities. Regardless of the developmental stage, neurosphere cultures presented large variability in the viability and proliferation rates during the initial 3-4 passages, but stabilized achieving significant expansion rates at passage 5 to 6. This was true also for all brain regions except cerebellar derived cultures that did not expand. Interestingly, the brain region of hfNSC derivation influences the expansion potential, being forebrain, cortex and WGE derived cells the most expandable compared to cerebellar. Short term expansion partially compromised the regional identity of cortical but not WGE cultures. Nevertheless, both expanded cultures were multipotent and kept the ability to differentiate to region specific mature neuronal phenotypes. Copyright © 2017 Elsevier Inc. All rights reserved.

  10. Spaceflight Activates Protein Kinase C Alpha Signaling and Modifies the Developmental Stage of Human Neonatal Cardiovascular Progenitor Cells.

    PubMed

    Baio, Jonathan; Martinez, Aida F; Bailey, Leonard; Hasaniya, Nahidh; Pecaut, Michael J; Kearns-Jonker, Mary

    2018-02-12

    Spaceflight impacts cardiovascular function in astronauts; however, its impact on cardiac development and the stem cells that form the basis for cardiac repair is unknown. Accordingly, further research is needed to uncover the potential relevance of such changes to human health. Using simulated microgravity (SMG) generated by two-dimensional clinorotation and culture aboard the International Space Station (ISS), we assessed the effects of mechanical unloading on human neonatal cardiovascular progenitor cell (CPC) developmental properties and signaling. Following 6-7 days of SMG and 12 days of ISS culture, we analyzed changes in gene expression. Both environments induced the expression of genes that are typically associated with an earlier state of cardiovascular development. To understand the mechanism by which such changes occurred, we assessed the expression of mechanosensitive small RhoGTPases in SMG-cultured CPCs and observed decreased levels of RHOA and CDC42. Given the effect of these molecules on intracellular calcium levels, we evaluated changes in noncanonical Wnt/calcium signaling. After 6-7 days under SMG, CPCs exhibited elevated levels of WNT5A and PRKCA. Similarly, ISS-cultured CPCs exhibited elevated levels of calcium handling and signaling genes, which corresponded to protein kinase C alpha (PKCα), a calcium-dependent protein kinase, activation after 30 days. Akt was activated, whereas phosphorylated extracellular signal-regulated kinase levels were unchanged. To explore the effect of calcium induction in neonatal CPCs, we activated PKCα using hWnt5a treatment on Earth. Subsequently, early cardiovascular developmental marker levels were elevated. Transcripts induced by SMG and hWnt5a-treatment are expressed within the sinoatrial node, which may represent embryonic myocardium maintained in its primitive state. Calcium signaling is sensitive to mechanical unloading and directs CPC developmental properties. Further research both in space and on Earth may help refine the use of CPCs in stem cell-based therapies and highlight the molecular events of development.

  11. Comparison of culture media, simplate, and petrifilm for enumeration of yeasts and molds in food.

    PubMed

    Taniwaki, M H; Silva, N; Banhe, A A; Iamanaka, B T

    2001-10-01

    The efficacy of three culture media, dichloran rose bengal chloramphenicol (DRBC), dichloran 18% glycerol agar (DG18), and potato dextrose agar (PDA) supplemented with two antibiotics, were compared with the Simplate and Petrifilm techniques for mold and yeast enumeration. The following foods were analyzed: corn meal, wheat flour, cassava flour, bread crumbs, whole meal, sliced bread, ground peanuts, mozzarella cheese, grated parmesan cheese, cheese rolls, orange juice, pineapple pulp, pineapple cake, and mushroom in conserve. Correlation coefficients of DRBC versus PDA and DG18 for recovering total mold and yeast counts from the composite of 14 foods indicated that the three media were generally equivalent. Correlation coefficients for Petrifilm versus culture media were acceptable, although not as good as between culture media. Correlation coefficients of Simplate versus DRBC, DG18, PDA, and Petrifilm for recovering total yeasts and molds from a composite of 11 foods demonstrated that there was no equivalence between the counts obtained by Simplate and other culture media and Petrifilm, with significant differences observed for the most foods analyzed.

  12. Ras Family GTPases Control Growth of Astrocyte Processes

    PubMed Central

    Kalman, Daniel; Gomperts, Stephen N.; Hardy, Stephen; Kitamura, Marina; Bishop, J. Michael

    1999-01-01

    Astrocytes in neuron-free cultures typically lack processes, although they are highly process-bearing in vivo. We show that basic fibroblast growth factor (bFGF) induces cultured astrocytes to grow processes and that Ras family GTPases mediate these morphological changes. Activated alleles of rac1 and rhoA blocked and reversed bFGF effects when introduced into astrocytes in dissociated culture and in brain slices using recombinant adenoviruses. By contrast, dominant negative (DN) alleles of both GTPases mimicked bFGF effects. A DN allele of Ha-ras blocked bFGF effects but not those of Rac1-DN or RhoA-DN. Our results show that bFGF acting through c-Ha-Ras inhibits endogenous Rac1 and RhoA GTPases thereby triggering astrocyte process growth, and they provide evidence for the regulation of this cascade in vivo by a yet undetermined neuron-derived factor. PMID:10233170

  13. Cross-cultural validity of standardized motor development screening and assessment tools: a systematic review.

    PubMed

    Mendonça, Bianca; Sargent, Barbara; Fetters, Linda

    2016-12-01

    To investigate whether standardized motor development screening and assessment tools that are used to evaluate motor abilities of children aged 0 to 2 years are valid in cultures other than those in which the normative sample was established. This was a systematic review in which six databases were searched. Studies were selected based on inclusion/exclusion criteria and appraised for evidence level and quality. Study variables were extracted. Twenty-three studies representing six motor development screening and assessment tools in 16 cultural contexts met the inclusion criteria: Alberta Infant Motor Scale (n=7), Ages and Stages Questionnaire, 3rd edition (n=2), Bayley Scales of Infant and Toddler Development, 3rd edition (n=8), Denver Developmental Screening Test, 2nd edition (n=4), Harris Infant Neuromotor Test (n=1), and Peabody Developmental Motor Scales, 2nd edition (n=1). Thirteen studies found significant differences between the cultural context and normative sample. Two studies established reliability and/or validity of standardized motor development assessments in high-risk infants from different cultural contexts. Five studies established new population norms. Eight studies described the cross-cultural adaptation of a standardized motor development assessment. Standardized motor development assessments have limited validity in cultures other than that in which the normative sample was established. Their use can result in under- or over-referral for services. © 2016 Mac Keith Press.

  14. A cross-cultural comparison of the developmental evolution of expertise in diabetes self-management.

    PubMed

    Shimizu, Yasuko; Paterson, Barbara L

    2007-11-01

    The authors compare the findings of two research studies, one conducted in Japan and the other in Canada, about the developmental evolution of self-management of diabetes. In this article, the authors identify the similarities and differences that exist in the research data, proposing that the differences are situated in the different cultural perspectives of self-management that exist in both countries. Researchers have acknowledged that self-management has cultural dimensions. Despite this, however, there are few studies that have provided a cross-cultural comparison of the experience of self-management among different cultural groups. The authors conducted a critical comparative analysis of two models of developing expertise in diabetes self-management. The review included an analysis of the cultural meanings of the various terms and the underlying assumptions of both models. The models shared many similarities; however, their differences were identified, such as the meaning and interpretation of various words or experiences, and shaped by the culturally bound perspectives of self and health. The findings serve as a caution to imposing ethnocentric views and interpretations in diabetes care. In addition, they remind us about the importance of asking people with diabetes about what they understand, desire and understand. The findings challenge nurses to reflect on how the development of self-management of diabetes in various national contexts is influenced by health care practices that focus on control or harmony.

  15. Multicultural and Anti-Racist Education: The Developmental Rationale and Practical Implications.

    ERIC Educational Resources Information Center

    Mock, Karen R.

    This paper examines the meaning of multicultural education as a means of insuring that students feel comfortable in more than one cultural context. It outlines stages of child development and theories of learning behavior, especially as they relate to the development of cultural and racial attitudes. This paper examines behavioral development in…

  16. The Spirit of Culture: Applying Cultural Competency to Strength-Based Youth Development.

    ERIC Educational Resources Information Center

    Lucero, Maria Guajardo

    Applying the research-based developmental assets list of 40 positive relationships, experiences, and values as a framework for positive youth development provides communities with a set of factors associated with increased healthy behaviors and fewer high-risk behaviors. Beginning with a question regarding the applicability of this model for youth…

  17. Transforming Education Research through Open Video Data Sharing

    ERIC Educational Resources Information Center

    Gilmore, Rick O.; Adolph, Karen E.; Millman, David S.; Gordon, Andrew

    2016-01-01

    Open data sharing promises to accelerate the pace of discovery in the developmental and learning sciences, but significant technical, policy, and cultural barriers have limited its adoption. As a result, most research on learning and development remains shrouded in a culture of isolation. Data sharing is the rare exception (Gilmore, 2016). Many…

  18. Identifying developmental toxicity pathways for a subset of ToxCast chemicals using human embryonic stem cells and metabolomics

    EPA Science Inventory

    Metabolomics analysis was performed on the supernatant of human embryonic stem (hES) cell cultures exposed to a blinded subset of 11 chemicals selected from the chemical library of EPA's ToxCast™ chemical screening and prioritization research project. Metabolites from hES cultur...

  19. Development of the Other-Race Effect during Infancy: Evidence toward Universality?

    ERIC Educational Resources Information Center

    Kelly, David J.; Liu, Shaoying; Lee, Kang; Quinn, Paul C.; Pascalis, Olivier; Slater, Alan M.; Ge, Liezhong

    2009-01-01

    The other-race effect in face processing develops within the first year of life in Caucasian infants. It is currently unknown whether the developmental trajectory observed in Caucasian infants can be extended to other cultures. This is an important issue to investigate because recent findings from cross-cultural psychology have suggested that…

  20. College Students' Life Experiences in Korea and in the U.S.

    ERIC Educational Resources Information Center

    Kang, Sunwoo; Youn, Gahyun; Stilwell, William E.

    2014-01-01

    Even though college students may be faced with developmental life tasks that might lead them to cognitive and emotional growth, little is known about how college students' life experiences are related to their cultural values. Understanding college students' life across culture requires both exploratory and confirmatory approaches to examine (1)…

  1. Socio-Cultural Theories of Cognitive Development: Implications for Teaching Theory in the Visual Arts.

    ERIC Educational Resources Information Center

    Fielding, Rob

    1989-01-01

    Explicates the socio-cultural developmental theories of Vygotsky and Feuerstein which advocate teacher mediated learning in order to stimulate and accelerate development. Implications for art education include the need for the teacher to become involved in the enculturation of the child into the thinking processes and conceptual organization of…

  2. Trade-Offs in the Study of Culture and Development: Theories, Methods, and Values.

    ERIC Educational Resources Information Center

    Rothbaum, Fred; Pott, Martha; Azuma, Hiroshi; Miyake, Kazuo; Weisz, John

    2000-01-01

    Notes that commentators unanimously support Rothbaum et al.'s general orientation to culture and development and their developmental pathways. Views commentators' suggestions as relating to trade-offs: between theories that highlight generalization or exceptions; between methods that rely on one-, two-, or multiculture studies; and between values…

  3. Sex and Age Differences in Achievement Goal Orientations in Turkish Adolescents

    ERIC Educational Resources Information Center

    Sahin, Ertugrul; Topkaya, Nursel; Kürkçü, Recep

    2016-01-01

    Culture plays an important role in the achievement goal orientations of students, which may vary as they progress through their lifespan. However, research examining achievement goal orientations in the Turkish cultural context is scarce. Based on contextual and developmental theories, the aim of this study was to examine sex and age differences…

  4. Preserving Guam's Culture with Culturally Responsive Children's Stories

    ERIC Educational Resources Information Center

    Jackson, Marilyn Malloy; Heath, Melissa Allen

    2017-01-01

    Regardless of where schools are located, teachers face the challenge of teaching and mentoring children, not only in academic achievement but in social emotional development. When faced with challenges, young children are especially vulnerable because they lack the life experience and developmental maturity to adequately cope. Relying on the lead…

  5. CAKES (Cultural Awareness and Knowledge Exchange Scheme): A Holistic and Inclusive Approach to Supporting International Students

    ERIC Educational Resources Information Center

    McKay, Jane; O'Neill, Deborah; Petrakieva, Lina

    2018-01-01

    Transition support for international students has traditionally adopted deficit models which attempt to "fix" assumed academic literacy problems. This study explores a more culturally inclusive initiative which supported international students at a UK university in a holistic and developmental way. The initiative was delivered across an…

  6. Supporting Transitions: Cultural Connections for Adults with Autism Spectrum Disorders

    ERIC Educational Resources Information Center

    Greenberg, Aliza; Levinsky-Raskin, Sheri

    2017-01-01

    Museums are well-positioned to engage adults with autism and other developmental differences both as visitors and employees. This article recounts the Museum Access Consortium's process to design and implement the project Supporting Transitions: Cultural Connections for People with Autism with the goal of calling attention to a large opportunity…

  7. Changing Views of Culture and Behavior: An Interview with Robert Serpell

    ERIC Educational Resources Information Center

    Lockett, Charles T.

    2003-01-01

    Charles Lockett is an Assistant Professor in the School of Psychology at James Madison University, where he teaches developmental psychology as well as advanced topic courses in cultural psychology and the psychology of race and racism. A graduate of Howard University, Lockett credits Howard's Preparing Future Faculty Fellowship Program for his…

  8. Investigating the Mechanisms Underlying Neuronal Death in Ischemia Using In Vitro Oxygen-Glucose Deprivation: Potential Involvement of Protein SUMOylation

    PubMed Central

    CIMAROSTI, HELENA; HENLEY, JEREMY M.

    2012-01-01

    It is well established that brain ischemia can cause neuronal death via different signaling cascades. The relative importance and interrelationships between these pathways, however, remain poorly understood. Here is presented an overview of studies using oxygen-glucose deprivation of organotypic hippocampal slice cultures to investigate the molecular mechanisms involved in ischemia. The culturing techniques, setup of the oxygen-glucose deprivation model, and analytical tools are reviewed. The authors focus on SUMOylation, a posttranslational protein modification that has recently been implicated in ischemia from whole animal studies as an example of how these powerful tools can be applied and could be of interest to investigate the molecular pathways underlying ischemic cell death. PMID:19029060

  9. Towards a Fail-Safe Air Force Culture: Creating a Resilient Future While Avoiding Past Mistakes

    DTIC Science & Technology

    2011-02-16

    for either preventing catastrophic failures or in the event they occur. The Air Force Safety process often uses the ― Swiss Cheese ‖ model to...evaluate accidents. The image of holes in the protective cheese layers (proactive and reactive measures) lining up in such a way as to allow an accident is... cheese . More importantly, however, a HRO‘s focus is on ―the process of the slices lining up as each moment where one hole aligns with another

  10. Endogenous Bone Regeneration Is Dependent Upon a Dynamic Oxygen Event

    DTIC Science & Technology

    2014-01-01

    endosteum and nail epithelium (Fig. 2G ). By DPA 21 hypoxyprobe signaling is still apparent in osteoblasts in the newly formed trabecular bone, but to a...affiliated with cells that surround the vascular lumen (Fig. 2G ’). This signal within the distal tip of P3 expands at DPA 21, associated...vivo (Fig. 3G -I). Digit slices were cultured at 21% oxygen and 5% CO2. Bone formation was assessed using Alizarin red staining. Bone morphology is

  11. Differential Alterations in Excitatory and Inhibitory Networks Involving Dentate Granule Cells Following Chronic Treatment with Distinct Classes of NMDAR Antagonists in Hippocampal Slice Cultures

    DTIC Science & Technology

    2010-03-08

    1992; Jung and McNaughton, 1993); (2) low incidence of recurrent excitatory synapses between granule cells (Molnar and Nadler, 1999; Okazaki et al...neurons, dentate granule cells have a relatively more negative resting membrane potential and exhibit low-frequency firing (Staley et al., 1992; Jung ...inhibition plays a dual role in brain function and possibly seizure occurrence through balancing excitation and synchronizing neuronal firing. An

  12. Brief Report: "Quick and (Not So) Dirty" Assessment of Change in Autism--Cross-Cultural Reliability of the Developmental Disabilities CGAS and the OSU Autism CGI

    ERIC Educational Resources Information Center

    Choque Olsson, Nora; Bölte, Sven

    2014-01-01

    There are few evaluated economic tools to assess change in autism. This study examined the inter-rater reliability of the Developmental Disabilities Children's Global Assessment Scale (DD-CGAS), and the OSU Autism Clinical Global Impression (OSU Autism CGI) in a European setting. Using these scales, 16 clinicians with multidisciplinary…

  13. Recent Insights Into Molecular Mechanisms of Propofol-Induced Developmental Neurotoxicity: Implications for the Protective Strategies.

    PubMed

    Bosnjak, Zeljko J; Logan, Sarah; Liu, Yanan; Bai, Xiaowen

    2016-11-01

    Mounting evidence has demonstrated that general anesthetics could induce developmental neurotoxicity, including acute widespread neuronal cell death, followed by long-term memory and learning abnormalities. Propofol is a commonly used intravenous anesthetic agent for the induction and maintenance of anesthesia and procedural and critical care sedation in children. Compared with other anesthetic drugs, little information is available on its potential contributions to neurotoxicity. Growing evidence from multiple experimental models showed a similar neurotoxic effect of propofol as observed in other anesthetic drugs, raising serious concerns regarding pediatric propofol anesthesia. The aim of this review is to summarize the current findings of propofol-induced developmental neurotoxicity. We first present the evidence of neurotoxicity from animal models, animal cell culture, and human stem cell-derived neuron culture studies. We then discuss the mechanism of propofol-induced developmental neurotoxicity, such as increased cell death in neurons and oligodendrocytes, dysregulation of neurogenesis, abnormal dendritic development, and decreases in neurotrophic factor expression. Recent findings of complex mechanisms of propofol action, including alterations in microRNAs and mitochondrial fission, are discussed as well. An understanding of the toxic effect of propofol and the underlying mechanisms may help to develop effective novel protective or therapeutic strategies for avoiding the neurotoxicity in the developing human brain.

  14. A dysmorphology score system for assessing embryo abnormalities in rat whole embryo culture.

    PubMed

    Zhang, Cindy X; Danberry, Tracy; Jacobs, Mary Ann; Augustine-Rauch, Karen

    2010-12-01

    The rodent whole embryo culture (WEC) system is a well-established model for characterizing developmental toxicity of test compounds and conducting mechanistic studies. Laboratories have taken various approaches in describing type and severity of developmental findings of organogenesis-stage rodent embryos, but the Brown and Fabro morphological score system is commonly used as a quantitative approach. The associated score criteria is based upon developmental stage and growth parameters, where a series of embryonic structures are assessed and assigned respective scores relative to their gestational stage, with a Total Morphological Score (TMS) assigned to the embryo. This score system is beneficial because it assesses a series of stage-specific anatomical landmarks, facilitating harmonized evaluation across laboratories. Although the TMS provides a quantitative approach to assess growth and determine developmental delay, it is limited to its ability to identify and/or delineate subtle or structure-specific abnormalities. Because of this, the TMS may not be sufficiently sensitive for identifying compounds that induce structure or organ-selective effects. This study describes a distinct morphological score system called the "Dysmorphology Score System (DMS system)" that has been developed for assessing gestation day 11 (approximately 20-26 somite stage) rat embryos using numerical scores to differentiate normal from abnormal morphology and define the respective severity of dysmorphology of specific embryonic structures and organ systems. This method can also be used in scoring mouse embryos of the equivalent developmental stage. The DMS system enhances capabilities to rank-order compounds based upon teratogenic potency, conduct structure- relationships of chemicals, and develop statistical prediction models to support abbreviated developmental toxicity screens. © 2010 Wiley-Liss, Inc.

  15. Developmental Competence and Epigenetic Profile of Porcine Embryos Produced by Two Different Cloning Methods.

    PubMed

    Liu, Ying; Lucas-Hahn, Andrea; Petersen, Bjoern; Li, Rong; Hermann, Doris; Hassel, Petra; Ziegler, Maren; Larsen, Knud; Niemann, Heiner; Callesen, Henrik

    2017-06-01

    The "Dolly" based cloning (classical nuclear transfer, [CNT]) and the handmade cloning (HMC) are methods that are nowadays routinely used for somatic cloning of large domestic species. Both cloning protocols share several similarities, but differ with regard to the required in vitro culture, which in turn results in different time intervals until embryo transfer. It is not yet known whether the differences between cloned embryos from the two protocols are due to the cloning methods themselves or the in vitro culture, as some studies have shown detrimental effects of in vitro culture on conventionally produced embryos. The goal of this study was to unravel putative differences between two cloning methods, with regard to developmental competence, expression profile of a panel of developmentally important genes and epigenetic profile of porcine cloned embryos produced by either CNT or HMC, either with (D5 or D6) or without (D0) in vitro culture. Embryos cloned by these two methods had a similar morphological appearance on D0, but displayed different cleavage rates and different quality of blastocysts, with HMC embryos showing higher blastocyst rates (HMC vs. CNT: 35% vs. 10%, p < 0.05) and cell numbers per blastocyst (HMC vs. CNT: 31 vs. 23 on D5 and 42 vs. 18 on D6, p < 0.05) compared to CNT embryos. With regard to histone acetylation and gene expression, CNT and HMC derived cloned embryos were similar on D0, but differed on D6. In conclusion, both cloning methods and the in vitro culture may affect porcine embryo development and epigenetic profile. The two cloning methods essentially produce embryos of similar quality on D0 and after 5 days in vitro culture, but thereafter both histone acetylation and gene expression differ between the two types of cloned embryos.

  16. Proteomic Analysis of Grape Berry Cell Cultures Reveals that Developmentally Regulated Ripening Related Processes Can Be Studied Using Cultured Cells

    PubMed Central

    Sharathchandra, Ramaschandra G.; Stander, Charmaine; Jacobson, Dan; Ndimba, Bongani; Vivier, Melané A.

    2011-01-01

    Background This work describes a proteomics profiling method, optimized and applied to berry cell suspensions to evaluate organ-specific cultures as a platform to study grape berry ripening. Variations in berry ripening within a cluster(s) on a vine and in a vineyard are a major impediment towards complete understanding of the functional processes that control ripening, specifically when a characterized and homogenous sample is required. Berry cell suspensions could overcome some of these problems, but their suitability as a model system for berry development and ripening needs to be established first. Methodology/Principal Findings In this study we report on the proteomic evaluation of the cytosolic proteins obtained from synchronized cell suspension cultures that were established from callus lines originating from green, véraison and ripe Vitis vinifera berry explants. The proteins were separated using liquid phase IEF in a Microrotofor cell and SDS PAGE. This method proved superior to gel-based 2DE. Principal component analysis confirmed that biological and technical repeats grouped tightly and importantly, showed that the proteomes of berry cultures originating from the different growth/ripening stages were distinct. A total of twenty six common bands were selected after band matching between different growth stages and twenty two of these bands were positively identified. Thirty two % of the identified proteins are currently annotated as hypothetical. The differential expression profile of the identified proteins, when compared with published literature on grape berry ripening, suggested common trends in terms of relative abundance in the different developmental stages between real berries and cell suspensions. Conclusions The advantages of having suspension cultures that accurately mimic specific developmental stages are profound and could significantly contribute to the study of the intricate regulatory and signaling networks responsible for berry development and ripening. PMID:21379583

  17. Attachment-past and present. But what about the future?

    PubMed

    Keller, Heidi

    2008-12-01

    This essay acknowledges the seminal contribution of the founders of attachment theory and research as a paradigm shift in understanding the importance of the socioemotional foundation of developmental processes. However, it is surprising that attachment theory has been treated as a closed system by its students with remarkable resistance to change and further development of theory and methodology. Especially three dimensions are identified that would help to advance attachment theory as a valid conception also for the future. The context specificity of evolutionary theorizing, the different socialization goals and parenting strategies across cultures and the new insights in infant development need to be taken into consideration in order to develop a valid theory of socioemotional development as the culturally informed solution of universal developmental tasks.

  18. A shape-based statistical method to retrieve 2D TRUS-MR slice correspondence for prostate biopsy

    NASA Astrophysics Data System (ADS)

    Mitra, Jhimli; Srikantha, Abhilash; Sidibé, Désiré; Martí, Robert; Oliver, Arnau; Lladó, Xavier; Ghose, Soumya; Vilanova, Joan C.; Comet, Josep; Meriaudeau, Fabrice

    2012-02-01

    This paper presents a method based on shape-context and statistical measures to match interventional 2D Trans Rectal Ultrasound (TRUS) slice during prostate biopsy to a 2D Magnetic Resonance (MR) slice of a pre-acquired prostate volume. Accurate biopsy tissue sampling requires translation of the MR slice information on the TRUS guided biopsy slice. However, this translation or fusion requires the knowledge of the spatial position of the TRUS slice and this is only possible with the use of an electro-magnetic (EM) tracker attached to the TRUS probe. Since, the use of EM tracker is not common in clinical practice and 3D TRUS is not used during biopsy, we propose to perform an analysis based on shape and information theory to reach close enough to the actual MR slice as validated by experts. The Bhattacharyya distance is used to find point correspondences between shape-context representations of the prostate contours. Thereafter, Chi-square distance is used to find out those MR slices where the prostates closely match with that of the TRUS slice. Normalized Mutual Information (NMI) values of the TRUS slice with each of the axial MR slices are computed after rigid alignment and consecutively a strategic elimination based on a set of rules between the Chi-square distances and the NMI leads to the required MR slice. We validated our method for TRUS axial slices of 15 patients, of which 11 results matched at least one experts validation and the remaining 4 are at most one slice away from the expert validations.

  19. Organotypic Cultures from the Adult CNS: A Novel Model to Study Demyelination and Remyelination Ex Vivo.

    PubMed

    Tan, Glaiza A; Furber, Kendra L; Thangaraj, Merlin P; Sobchishin, LaRhonda; Doucette, J Ronald; Nazarali, Adil J

    2018-01-01

    Experimental models of multiple sclerosis (MS) have significantly advanced our understanding of pathophysiology and therapeutic interventions. Although in vivo rodent models are considered to most closely represent the complex cellular and molecular disease states of the human central nervous system (CNS), these can be costly to maintain and require long timelines. Organotypic slice cultures maintain the cytotypic organization observed in the intact CNS, yet provide many of the experimental advantages of in vitro cell culture models. Cerebellar organotypic cultures have proven useful for studying myelination and remyelination, but this model has only been established using early postnatal tissue. This young brain tissue allows for neuro development ex vivo to mimic the 'mature' CNS; however, there are many differences between postnatal and adult organotypic cultures. This may be particularly relevant to MS, as a major barrier to myelin regeneration is age. This paper describes a modified protocol to study demyelination and remyelination in adult cerebellar tissue, which has been used to demonstrate neuroprotection with omega-3 fatty acids. Thus, adult cerebellar organotypic cultures provide a novel ex vivo platform for screening potential therapies in myelin degeneration and repair.

  20. Early Studies on Protoplast Isolation of Ludisia discolor, A Wild Orchid

    PubMed Central

    Poobathy, Ranjetta; Zakaria, Rahmad; Hamzah, Syed Mohd. Edzham Syed; Subramaniam, Sreeramanan

    2016-01-01

    The terrestrial Ludisia discolor, also referred to as the jewel orchid is prized for the quality of its leaves. L. discolor is known as a medicinal herb and is touted for its heat- and pathogen-resisting qualities. L. discolor is valuable in the production of both flavonoids and anthocyanins, antioxidants that are exalted in the health industry. Plant cell cultures have emerged as alternative sources of anthocyanin production. Plant protoplast cultures are used frequently in transient gene expression studies and in the establishment of callus and cell suspension cultures. Benefits of plant protoplast system include similarity to cells found in plant tissues, reproduction under controlled conditions, and prevention of masking of stress responses to previous handling techniques. A study was conducted to assess the amenability of the stem and leaves of L. discolor to protoplast isolation. The stem and leaf segments were weighed, sliced into thin layers, immersed in a digestion medium, washed and then cultured onto a recovery medium. Results indicated that the production of plant protoplasts from L. discolor may be viewed as an alternative in the generation of cell cultures and ultimately in the production of anthocyanins from the cell cultures. PMID:27965736

  1. Early Studies on Protoplast Isolation of Ludisia discolor, A Wild Orchid.

    PubMed

    Poobathy, Ranjetta; Zakaria, Rahmad; Hamzah, Syed Mohd Edzham Syed; Subramaniam, Sreeramanan

    2016-11-01

    The terrestrial Ludisia discolor , also referred to as the jewel orchid is prized for the quality of its leaves. L. discolor is known as a medicinal herb and is touted for its heat- and pathogen-resisting qualities. L. discolor is valuable in the production of both flavonoids and anthocyanins, antioxidants that are exalted in the health industry. Plant cell cultures have emerged as alternative sources of anthocyanin production. Plant protoplast cultures are used frequently in transient gene expression studies and in the establishment of callus and cell suspension cultures. Benefits of plant protoplast system include similarity to cells found in plant tissues, reproduction under controlled conditions, and prevention of masking of stress responses to previous handling techniques. A study was conducted to assess the amenability of the stem and leaves of L. discolor to protoplast isolation. The stem and leaf segments were weighed, sliced into thin layers, immersed in a digestion medium, washed and then cultured onto a recovery medium. Results indicated that the production of plant protoplasts from L. discolor may be viewed as an alternative in the generation of cell cultures and ultimately in the production of anthocyanins from the cell cultures.

  2. Slice regular functions of several Clifford variables

    NASA Astrophysics Data System (ADS)

    Ghiloni, R.; Perotti, A.

    2012-11-01

    We introduce a class of slice regular functions of several Clifford variables. Our approach to the definition of slice functions is based on the concept of stem functions of several variables and on the introduction on real Clifford algebras of a family of commuting complex structures. The class of slice regular functions include, in particular, the family of (ordered) polynomials in several Clifford variables. We prove some basic properties of slice and slice regular functions and give examples to illustrate this function theory. In particular, we give integral representation formulas for slice regular functions and a Hartogs type extension result.

  3. Modular electronics packaging system

    NASA Technical Reports Server (NTRS)

    Hunter, Don J. (Inventor)

    2001-01-01

    A modular electronics packaging system includes multiple packaging slices that are mounted horizontally to a base structure. The slices interlock to provide added structural support. Each packaging slice includes a rigid and thermally conductive housing having four side walls that together form a cavity to house an electronic circuit. The chamber is enclosed on one end by an end wall, or web, that isolates the electronic circuit from a circuit in an adjacent packaging slice. The web also provides a thermal path between the electronic circuit and the base structure. Each slice also includes a mounting bracket that connects the packaging slice to the base structure. Four guide pins protrude from the slice into four corresponding receptacles in an adjacent slice. A locking element, such as a set screw, protrudes into each receptacle and interlocks with the corresponding guide pin. A conduit is formed in the slice to allow electrical connection to the electronic circuit.

  4. Culture of bovine embryos on a polydimethylsiloxane (PDMS) microwell plate.

    PubMed

    Akagi, Satoshi; Hosoe, Misa; Matsukawa, Kazutsugu; Ichikawa, Akihiko; Tanikawa, Tamio; Takahashi, Seiya

    2010-08-01

    We fabricated a polydimethylsiloxane (PDMS)-based microwell plate (PDMS-MP) containing 100 microwells with a rounded bottom and examined whether it can be used for culture of individual in vitro fertilized (IVF) embryos or parthenogenetically activated zona-free embryos in cattle. In Experiment 1, we examined the in vitro developmental ability of IVF embryos cultured individually on PDMS-MP. After IVF, 20 embryos were transferred into 100 microl drops on PDMS-MP and cultured individually in each well of PDMS-MP (PDMS group). After 7 days of culture, the embryos in the PDMS group developed to the blastocyst stage at the same rate of those in the control group cultured in a group of 20 embryos without PDMS-MP. There were no differences in total number of cells and the ratio of inner cell mass to total cells between the PDMS and control groups. In Experiment 2, we examined the in vitro developmental ability of parthenogenetically activated zona-free bovine embryos cultured individually on PDMS-MP. The zona-free embryos were cultured individually in each well of a PDMS-MP or in each well produced by pressing a darning needle onto the bottom of a culture dish (WOW group). After 7 days of culture, the blastocyst formation rate and cell number of blastocysts in the PDMS group did not differ from those of the zona-intact embryos in the control group. Also, there were no differences in the blastocyst formation rate and cell number of blastocysts between the WOW and PDMS groups. These results suggest that the culture system using PDMS-MP is useful for individual embryos or zona-free embryos in cattle.

  5. Enhanced excitatory input to melanin concentrating hormone neurons during developmental period of high food intake is mediated by GABA.

    PubMed

    Li, Ying; van den Pol, Anthony N

    2009-12-02

    In contrast to the local axons of GABA neurons of the cortex and hippocampus, lateral hypothalamic neurons containing melanin concentrating hormone (MCH) and GABA send long axons throughout the brain and play key roles in energy homeostasis and mental status. In adults, MCH neurons maintain a hyperpolarized membrane potential and most of the synaptic input is inhibitory. In contrast, we found that developing MCH neurons received substantially more excitatory synaptic input. Based on gramicidin-perforated patch recordings in hypothalamic slices from MCH-green fluorescent protein transgenic mice, we found that GABA was the primary excitatory synaptic transmitter in embryonic and neonatal ages up to postnatal day 10. Surprisingly, glutamate assumed only a minor excitatory role, if any. GABA plays a complex role in developing MCH neurons, with its actions conditionally dependent on a number of factors. GABA depolarization could lead to an increase in spikes either independently or in summation with other depolarizing stimuli, or alternately, depending on the relative timing of other depolarizing events, could lead to shunting inhibition. The developmental shift from depolarizing to hyperpolarizing occurred later in the dendrites than in the cell body. Early GABA depolarization was based on a Cl(-)-dependent inward current. An interesting secondary depolarization in mature neurons that followed an initial hyperpolarization was based on a bicarbonate mechanism. Thus during the early developmental period when food consumption is high, MCH neurons are more depolarized than in the adult, and an increased level of excitatory synaptic input to these orexigenic cells is mediated by GABA.

  6. Enhanced excitatory input to MCH neurons during developmental period of high food intake is mediated by GABA

    PubMed Central

    Li, Ying; van den Pol, Anthony N.

    2010-01-01

    In contrast to the local axons of GABA neurons of the cortex and hippocampus, lateral hypothalamic neurons containing melanin concentrating hormone (MCH) and GABA send long axons throughout the brain and play key roles in energy homeostasis and mental status. In adults, MCH neurons maintain a hyperpolarized membrane potential and most of the synaptic input is inhibitory. In contrast, we found that developing MCH neurons received substantially more excitatory synaptic input. Based on gramicidicin-perforated patch recordings in hypothalamic slices from MCH-GFP transgenic mice, we found that GABA was the primary excitatory synaptic transmitter in embryonic and neonatal ages up to postnatal day 10. Surprisingly, glutamate assumed only a minor excitatory role, if any. GABA plays a complex role in developing MCH neurons, with its actions conditionally dependent on a number of factors. GABA depolarization could lead to an increase in spikes either independently or in summation with other depolarizing stimuli, or alternately, depending on the relative timing of other depolarizing events, could lead to shunting inhibition. The developmental shift from depolarizing to hyperpolarizing occurred later in the dendrites than in the cell body. Early GABA depolarization was based on a Cl− dependent inward current. An interesting secondary depolarization in mature neurons that followed an initial hyperpolarization was based on a bicarbonate mechanism. Thus during the early developmental period when food consumption is high, MCH neurons are more depolarized than in the adult, and an increased level of excitatory synaptic input to these orexigenic cells is mediated by GABA. PMID:19955372

  7. Developmental Changes in Short-Term Plasticity at the Rat Calyx of Held Synapse

    PubMed Central

    Crins, Tom T. H.; Rusu, Silviu I.; Rodríguez-Contreras, Adrian; Borst, J. Gerard G.

    2015-01-01

    The calyx of Held synapse of the medial nucleus of the trapezoid body functions as a relay synapse in the auditory brainstem. In vivo recordings have shown that this synapse displays low release probability and that the average size of synaptic potentials does not depend on recent history. We used a ventral approach to make in vivo extracellular recordings from the calyx of Held synapse in rats aged postnatal day 4 (P4) to P29 to study the developmental changes that allow this synapse to function as a relay. Between P4 and P8, we observed evidence for the presence of large short-term depression, which was counteracted by short-term facilitation at short intervals. Major changes occurred in the last few days before the onset of hearing for air-borne sounds, which happened at P13. The bursting pattern changed into a primary-like pattern, the amount of depression and facilitation decreased strongly, and the decay of facilitation became much faster. Whereas short-term plasticity was the most important cause of variability in the size of the synaptic potentials in immature animals, its role became minor around hearing onset and afterward. Similar developmental changes were observed during stimulation experiments both in brain slices and in vivo following cochlear ablation. Our data suggest that the strong reduction in release probability and the speedup of the decay of synaptic facilitation that happen just before hearing onset are important events in the transformation of the calyx of Held synapse into an auditory relay synapse. PMID:21832200

  8. Slice profile effects in 2D slice-selective MRI of hyperpolarized nuclei.

    PubMed

    Deppe, Martin H; Teh, Kevin; Parra-Robles, Juan; Lee, Kuan J; Wild, Jim M

    2010-02-01

    This work explores slice profile effects in 2D slice-selective gradient-echo MRI of hyperpolarized nuclei. Two different sequences were investigated: a Spoiled Gradient Echo sequence with variable flip angle (SPGR-VFA) and a balanced Steady-State Free Precession (SSFP) sequence. It is shown that in SPGR-VFA the distribution of flip angles across the slice present in any realistically shaped radiofrequency (RF) pulse leads to large excess signal from the slice edges in later RF views, which results in an undesired non-constant total transverse magnetization, potentially exceeding the initial value by almost 300% for the last RF pulse. A method to reduce this unwanted effect is demonstrated, based on dynamic scaling of the slice selection gradient. SSFP sequences with small to moderate flip angles (<40 degrees ) are also shown to preserve the slice profile better than the most commonly used SPGR sequence with constant flip angle (SPGR-CFA). For higher flip angles, the slice profile in SSFP evolves in a manner similar to SPGR-CFA, with depletion of polarization in the center of the slice. Copyright 2009 Elsevier Inc. All rights reserved.

  9. Globalisation and Localisation Enmeshed--Towards a Framework for the Development of Guidance Curriculum in Hong Kong

    ERIC Educational Resources Information Center

    Luk-Fong, Pattie Yuk Yee

    2005-01-01

    Hong Kong's 150 years of colonisation by the British is a classic example of how globalisation comes into direct contact with a local Chinese culture and creates cultural hybridities. The paper presents a framework for developing a hybrid guidance curriculum, drawing together western traditions of developmental contextualism in guidance and…

  10. Using Mixed Methods Research to Examine the Benefits of Culturally Relevant Instruction on Latino Students' Writing Skills

    ERIC Educational Resources Information Center

    Murphy, Joel P.; Murphy, Shirley A.

    2016-01-01

    A convergent mixed methods research design addressed the extent of benefit obtained from reading culturally inclusive prompts (i.e., four brief essays written by Latino authors) to improve essay writing in a developmental (pre-college) English course. Participants were 45 Latino students who provided quantitative data. Chi square analysis showed…

  11. Parent-Child Socialization in Diverse Cultures. Annual Advances in Applied Developmental Psychology, Volume 5.

    ERIC Educational Resources Information Center

    Roopnarine, Jaipaul L., Ed.; Carter, D. Bruce, Ed.

    This collection of essays addresses the role of culture in the functioning of families and the socialization of children. Following an introduction by Irving Sigel, the 15 essays are: (1) "Parent-Child Interactions in Urban Indian Families in New Delhi: Are They Changing?" (Jaipaul Roopnarine and Ziarat Hossain); (2) "Chinese…

  12. Title VII Enhancement Project for Compartiendo Culturas/Sharing Cultures 1995-96. Research Report on Educational Programs.

    ERIC Educational Resources Information Center

    Houston Independent School District, TX. Dept. of Research and Evaluation.

    Compartiendo Culturas/Sharing Cultures, a Title VII Two-Way Developmental Bilingual Education Program at the Gary L. Herod Elementary School in the Houston Independent School District (Texas) was designed to end the isolation typically experienced by language minority students in traditional bilingual education and to provide language majority…

  13. The Effectiveness of Culturally-Based Social Stories to Increase Appropriate Behaviors of Children with Developmental Delays

    ERIC Educational Resources Information Center

    Hsu, Ning; Hammond, Helen; Ingalls, Lawrence

    2012-01-01

    The needs of students from culturally and linguistically diverse (CLD) backgrounds have become a growing concern in United States. As an even greater challenge, educators are looking at effective interventions to provide appropriate education for CLD students with disabilities. The Social Story technique is a practical behavioral intervention…

  14. The Volunteer and the Sannyasin: Archetypes of Retirement in America and India

    ERIC Educational Resources Information Center

    Savishinsky, Joel

    2004-01-01

    Cultures differ in their models of the life course, including the number and the content of the stages through which people are expected to pass. Among those societies that recognize a period of retirement, the developmental tasks and opportunities that older individuals face are shaped by cultural ideas about morality, spirituality, passion, and…

  15. Methodologies in Cultural-Historical Activity Theory: The Example of School-Based Development

    ERIC Educational Resources Information Center

    Postholm, May Britt

    2015-01-01

    Background and purpose: Relatively little research has been conducted on methodology within Cultural-Historical Activity Theory (CHAT). CHAT is mainly used as a framework for developmental processes. The purpose of this article is to discuss both focuses for research and research questions within CHAT and to outline methodologies that can be used…

  16. Symposium on Early Childhood Education (Urbana-Champaign, Illinois, November 9-10, 1997). Abstracts.

    ERIC Educational Resources Information Center

    Illinois Univ., Champaign. Coll. of Education.

    This book compiles abstracts of presentations from a symposium held in honor of Dr. Bernard Spodek, a leading scholar in early childhood education, on the occasion of his retirement. Topics addressed are as follows: (1) culture in early childhood curriculum; (2) developmental and cultural appropriateness; (3) kindergarten in Japan; (4) pressure…

  17. The Impact of Cultural Competence on the Moral Development of Student Affairs Professionals

    ERIC Educational Resources Information Center

    Cuyjet, Michael J.; Duncan, Angela D.

    2013-01-01

    As student affairs professionals, we are expected to meet students where they are developmentally and challenge and support them as they learn, develop, and grow into productive citizens. Since our institutions, and society as a whole, are becoming increasingly diverse, it is imperative that we acknowledge the numerous ways cultural background…

  18. A Qualitative Exploration of the Repatriation Experiences of US Third Culture Kids in College

    ERIC Educational Resources Information Center

    Smith, Virginia J.; Kearney, Kerri S.

    2016-01-01

    US third culture kids (TCKs), or those who spend a significant amount of their developmental years in other countries, often repatriate for college. However, the TCK population is typically invisible on US campuses, and understanding of their journeys is lacking. Using as lenses Erikson's stages of development (1963, 1968, 1997) and the…

  19. Finding the Authentic Self in a Communal Culture: Developmental Goals in Emerging Adulthood

    ERIC Educational Resources Information Center

    Scharf, Miri; Mayseless, Ofra

    2010-01-01

    Finding and cultivating a sense of authentic self is an important life goal for emerging adults. In collectivist cultures, youngsters might need to distance themselves to find and discover their authentic selves separate of the expectations of society and significant others. Creating an autonomous time bubble that focuses on the present allows…

  20. Developmental Consequences of Early Parenting Experiences: Self-Recognition and Self-Regulation in Three Cultural Communities

    ERIC Educational Resources Information Center

    Keller, Heidi; Yovsi, Relindis; Borke, Joern; Krtner, Joscha; Jensen, Henning; Papaligoura, Zaira

    2004-01-01

    This study relates parenting of 3-month-old children to children's self-recognition and self-regulation at 18 to 20 months. As hypothesized, observational data revealed differences in the sociocultural orientations of the 3 cultural samples' parenting styles and in toddlers' development of self-recognition and self-regulation. Children of…

  1. Developmentally Appropriate New Media Literacies: Supporting Cultural Competencies and Social Skills in Early Childhood Education

    ERIC Educational Resources Information Center

    Alper, Meryl

    2013-01-01

    Young children explore their world through manipulatives, playing with "technology" that may or may not be digital. To this end, I offer an exploration into how the existing framework of the New Media Literacies (NMLs) paradigm set forth by Henry Jenkins (2006) in "Confronting the Challenges of Participatory Culture: Media Education…

  2. From reassurance to irrelevance: adolescent psychology and homosexuality in America.

    PubMed

    Spurlock, J C

    2002-02-01

    American psychology by the 1920s contained a greater capacity for viewing some homosexual experiences as normal than most current historical literature suggests. Developmental psychologists agreed with psychiatrists that adult homosexuality was pathological, but they also agreed that adolescent sexual development included a homosexual phase. Until the late 1960s, developmental texts reassured parents and teachers that homosexual behavior among adolescents was transitory and quite normal. The psychiatric view of homosexuality as pathology came under attack after the middle of the century and eventually was abandoned. The developmental concern with a transitory homosexual phase disappeared gradually. This trend in psychology suggests underlying social and cultural changes.

  3. Primary cultures of astrocytes from fetal bovine brain.

    PubMed

    Ballarin, Cristina; Peruffo, Antonella

    2012-01-01

    We describe here a method to obtain primary cell cultures from the cerebral cortex and the hypothalamus of bovine fetuses. We report how tissue origin, developmental stages, and culture medium conditions influence cell differentiation and the prevalence of glial cells vs. neurons. We compare explants from early, middle, and late stages of development and two different fetal calf serum concentrations (1 and 10%) to identify the best conditions to obtain and grow viable astrocytes in culture. In addition, we describe how to cryopreserve and obtain viable cortical astrocytes from frozen fetal bovine brain samples.

  4. Cross-Cultural Strategies for Improving the Teaching, Training, and Mentoring Skills of Military Transition Team Advisors

    DTIC Science & Technology

    2010-04-01

    teaching— both in the classroom as well as field-based and individualized contexts. The developmental psychologist Jean Piaget is credited with the phrase...sponsor new theory and research in intercultural development . Bennett has been active in the intercultural field since 1967. Bennett and Hammer are well...known for their work with the Developmental Model of Intercultural Sensitivity (DMIS) and the Intercultural Development Inventory (IDI).    4

  5. Modeling the growth of Lactobacillus viridescens under non-isothermal conditions in vacuum-packed sliced ham.

    PubMed

    Silva, Nathália Buss da; Longhi, Daniel Angelo; Martins, Wiaslan Figueiredo; Laurindo, João Borges; Aragão, Gláucia Maria Falcão de; Carciofi, Bruno Augusto Mattar

    2017-01-02

    Lactic acid bacteria (LAB) are responsible for spoiling vacuum-packed meat products, such as ham. Since the temperature is the main factor affecting the microbial dynamic, the use of mathematical models describing the microbial behavior into a non-isothermal environment can be very useful for predicting food shelf life. In this study, the growth of Lactobacillus viridescens was measured in vacuum-packed sliced ham under non-isothermal conditions, and the predictive ability of primary (Baranyi and Roberts, 1994) and secondary (Square Root) models were assessed using parameters estimated in MRS culture medium under isothermal conditions (between 4 and 30°C). Fresh ham piece was sterilized, sliced, inoculated, vacuum-packed, and stored in a temperature-controlled incubator at five different non-isothermal conditions (between 4 and 25°C) and one isothermal condition (8°C). The mathematical models obtained in MRS medium were assessed by comparing predicted values with L. viridescens growth data in vacuum-packed ham. Its predictive ability was assessed through statistical indexes, with good results (bias factor between 0.95 and 1.03; accuracy factor between 1.04 and 1.07, and RMSE between 0.76 and 1.33), especially in increasing temperature, which predictions were safe. The model parameters obtained from isothermal growth data in MRS medium enabled to estimate the shelf life of a commercial ham under non-isothermal conditions in the temperature range analyzed. Copyright © 2016 Elsevier B.V. All rights reserved.

  6. Xenon Protects against Blast-Induced Traumatic Brain Injury in an In Vitro Model.

    PubMed

    Campos-Pires, Rita; Koziakova, Mariia; Yonis, Amina; Pau, Ashni; Macdonald, Warren; Harris, Katie; Edge, Christopher J; Franks, Nicholas P; Mahoney, Peter F; Dickinson, Robert

    2018-04-15

    The aim of this study was to evaluate the neuroprotective efficacy of the inert gas xenon as a treatment for patients with blast-induced traumatic brain injury in an in vitro laboratory model. We developed a novel blast traumatic brain injury model using C57BL/6N mouse organotypic hippocampal brain-slice cultures exposed to a single shockwave, with the resulting injury quantified using propidium iodide fluorescence. A shock tube blast generator was used to simulate open field explosive blast shockwaves, modeled by the Friedlander waveform. Exposure to blast shockwave resulted in significant (p < 0.01) injury that increased with peak-overpressure and impulse of the shockwave, and which exhibited a secondary injury development up to 72 h after trauma. Blast-induced propidium iodide fluorescence overlapped with cleaved caspase-3 immunofluorescence, indicating that shock-wave-induced cell death involves apoptosis. Xenon (50% atm) applied 1 h after blast exposure reduced injury 24 h (p < 0.01), 48 h (p < 0.05), and 72 h (p < 0.001) later, compared with untreated control injury. Xenon-treated injured slices were not significantly different from uninjured sham slices at 24 h and 72 h. We demonstrate for the first time that xenon treatment after blast traumatic brain injury reduces initial injury and prevents subsequent injury development in vitro. Our findings support the idea that xenon may be a potential first-line treatment for those with blast-induced traumatic brain injury.

  7. Higher transport and metabolism of glucose in astrocytes compared with neurons: a multiphoton study of hippocampal and cerebellar tissue slices.

    PubMed

    Jakoby, Patrick; Schmidt, Elke; Ruminot, Iván; Gutiérrez, Robin; Barros, L Felipe; Deitmer, Joachim W

    2014-01-01

    Glucose is the most important energy substrate for the brain, and its cellular distribution is a subject of great current interest. We have employed fluorescent glucose probes, the 2-deoxy-D-glucose derivates 6- and 2-([N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl) amino]-2-deoxy-D-glucose) (2-NBDG), to measure transport and metabolism of glucose in acute slices of mouse hippocampus and cerebellum. In the hippocampus, 6-NBDG, which is not metabolized and hence indicates glucose transport, was taken up faster in astrocyte-rich layers (Stratum radiatum [S.r.], Stratum oriens [S.o.]) than in pyramidal cells. Metabolizable 2-NBDG showed larger signals in S.r. and S.o. than in Stratum pyramidale, suggesting faster glucose utilization rate in the astrocyte versus the neuronal compartment. Similarly, we found higher uptake and temperature-sensitive metabolism of 2-NBDG in Bergmann glia when compared with adjacent Purkinje neurons of cerebellar slices. A comparison between 6-NBDG transport and glucose transport in cultured cells using a fluorescence resonance energy transfer nanosensor showed that relative to glucose, 6-NBDG is transported better by neurons than by astrocytes. These results indicate that the preferential transport and metabolism of glucose by glial cells versus neurons proposed for the hippocampus and cerebellum by ourselves (in vitro) and for the barrel cortex by Chuquet et al. (in vivo) is more pronounced than anticipated.

  8. Attenuation of Lipopolysaccharide-Induced Lung Vascular Stiffening by Lipoxin Reduces Lung Inflammation

    PubMed Central

    Meng, Fanyong; Mambetsariev, Isa; Tian, Yufeng; Beckham, Yvonne; Meliton, Angelo; Leff, Alan; Gardel, Margaret L.; Allen, Michael J.; Birukov, Konstantin G.

    2015-01-01

    Reversible changes in lung microstructure accompany lung inflammation, although alterations in tissue micromechanics and their impact on inflammation remain unknown. This study investigated changes in extracellular matrix (ECM) remodeling and tissue stiffness in a model of LPS-induced inflammation and examined the role of lipoxin analog 15-epi-lipoxin A4 (eLXA4) in the reduction of stiffness-dependent exacerbation of the inflammatory process. Atomic force microscopy measurements of live lung slices were used to directly measure local tissue stiffness changes induced by intratracheal injection of LPS. Effects of LPS on ECM properties and inflammatory response were evaluated in an animal model of LPS-induced lung injury, live lung tissue slices, and pulmonary endothelial cell (EC) culture. In vivo, LPS increased perivascular stiffness in lung slices monitored by atomic force microscopy and stimulated expression of ECM proteins fibronectin, collagen I, and ECM crosslinker enzyme, lysyl oxidase. Increased stiffness and ECM remodeling escalated LPS-induced VCAM1 and ICAM1 expression and IL-8 production by lung ECs. Stiffness-dependent exacerbation of inflammatory signaling was confirmed in pulmonary ECs grown on substrates with high and low stiffness. eLXA4 inhibited LPS-increased stiffness in lung cross sections, attenuated stiffness-dependent enhancement of EC inflammatory activation, and restored lung compliance in vivo. This study shows that increased local vascular stiffness exacerbates lung inflammation. Attenuation of local stiffening of lung vasculature represents a novel mechanism of lipoxin antiinflammatory action. PMID:24992633

  9. Early group bias in the Faroe Islands: Cultural variation in children's group-based reasoning.

    PubMed

    Schug, Mariah G; Shusterman, Anna; Barth, Hilary; Patalano, Andrea L

    2016-01-01

    Recent developmental research demonstrates that group bias emerges early in childhood. However, little is known about the extent to which bias in minimal (i.e., arbitrarily assigned) groups varies with children's environment and experience, and whether such bias is universal across cultures. In this study, the development of group bias was investigated using a minimal groups paradigm with 46 four- to six-year-olds from the Faroe Islands. Children observed in-group and out-group members exhibiting varying degrees of prosocial behaviour (egalitarian or stingy sharing). Children did not prefer their in-group in the pretest, but a pro-in-group and anti-out-group sentiment emerged in both conditions in the posttest. Faroese children's response patterns differ from those of American children [Schug, M. G., Shusterman, A., Barth, H., & Patalano, A. L. (2013). Minimal-group membership influences children's responses to novel experience with group members. Developmental Science, 16(1), 47-55], suggesting that intergroup bias shows cultural variation even in a minimal groups context.

  10. Recapitulating cortical development with organoid culture in vitro and modeling abnormal spindle-like (ASPM related primary) microcephaly disease.

    PubMed

    Li, Rui; Sun, Le; Fang, Ai; Li, Peng; Wu, Qian; Wang, Xiaoqun

    2017-11-01

    The development of a cerebral organoid culture in vitro offers an opportunity to generate human brain-like organs to investigate mechanisms of human disease that are specific to the neurogenesis of radial glial (RG) and outer radial glial (oRG) cells in the ventricular zone (VZ) and subventricular zone (SVZ) of the developing neocortex. Modeling neuronal progenitors and the organization that produces mature subcortical neuron subtypes during early stages of development is essential for studying human brain developmental diseases. Several previous efforts have shown to grow neural organoid in culture dishes successfully, however we demonstrate a new paradigm that recapitulates neocortical development process with VZ, OSVZ formation and the lamination organization of cortical layer structure. In addition, using patient-specific induced pluripotent stem cells (iPSCs) with dysfunction of the Aspm gene from a primary microcephaly patient, we demonstrate neurogenesis defects result in defective neuronal activity in patient organoids, suggesting a new strategy to study human developmental diseases in central nerve system.

  11. Reproducible and controllable light induction of in vitro fruiting of the white-rot basidiomycete Pleurotus ostreatus.

    PubMed

    Arjona, Davinia; Aragón, Carlos; Aguilera, José Antonio; Ramírez, Lucía; Pisabarro, Antonio G

    2009-05-01

    Fruiting is a crucial developmental process in basidiomycetes yet the genetic and molecular factors that control it are not yet fully understood. The search for fruiting inducers is of major relevance for both basic research and for their use in industrial applications. In this paper, an efficient and reproducible protocol for controlled fruiting induction of Pleurotus ostreatus growing on synthetic medium is described. The protocol is based on the control of light intensity and photoperiod and permits the life cycle for this fungus to be completed in less than two weeks. The fruiting bodies produced by this method release fertile spores after 4-5 d of culture. Our results indicate that fruiting induction is solely dependent on the illumination regime and that it occurs long before the available nutrients are depleted in the culture. This protocol will greatly facilitate molecular and developmental biology research in this fungus as it avoids the need for complex culture media based on lignocellulosic materials or the use of chemical inducers.

  12. Relationship Between Child and Maternal Sleep: A Developmental and Cross-Cultural Comparison.

    PubMed

    Mindell, Jodi A; Sadeh, Avi; Kwon, Robert; Goh, Daniel Y T

    2015-08-01

    The aim of this study was to assess the relationship between young children's sleep and maternal sleep from both a developmental and a cross-cultural perspective. Mothers of 10,085 young children completed the Brief Infant/Child Sleep Questionnaire and the Pittsburgh Sleep Quality Index. Overall, there were significant relationships between maternal and child sleep for bedtime, waketime, number of night wakings, and total nighttime sleep time across ages and cultures, although these relationships were stronger with younger children than preschool-aged children. Mothers report that their child's sleep pattern significantly impacts their sleep and daytime function, and they do not feel confident in managing their child's sleep pattern. Thus, interventions to improve children's sleep and develop good sleep habits, especially in early childhood, are likely to improve the quality of life of the whole family. © The Author 2015. Published by Oxford University Press on behalf of the Society of Pediatric Psychology. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  13. A constructivist theoretical proposition of cultural competence development in nursing.

    PubMed

    Blanchet Garneau, Amélie; Pepin, Jacinthe

    2015-11-01

    Cultural competence development in healthcare professions is considered an essential condition to promote quality and equity in healthcare. Even if cultural competence has been recognized as continuous, evolutionary, dynamic, and developmental by most researchers, current models of cultural competence fail to present developmental levels of this competence. These models have also been criticized for their essentialist perspective of culture and their limited application to competency-based approach programs. To our knowledge, there have been no published studies, from a constructivist perspective, of the processes involved in the development of cultural competence among nurses and undergraduate student nurses. The purpose of this study was to develop a theoretical proposition of cultural competence development in nursing from a constructivist perspective. We used a grounded theory design to study cultural competence development among nurses and student nurses in a healthcare center located in a culturally diverse urban area. Data collection involved participant observation and semi-structured interviews with 24 participants (13 nurses and 11 students) working in three community health settings. The core category, 'learning to bring the different realities together to provide effective care in a culturally diverse context', was constructed using inductive qualitative data analysis. This core category encompasses three dimensions of cultural competence: 'building a relationship with the other', 'working outside the usual practice framework', and 'reinventing practice in action.' The resulting model describes the concurrent evolution of these three dimensions at three different levels of cultural competence development. This study reveals that clinical experience and interactions between students or nurses and their environment both contribute significantly to cultural competence development. The resulting theoretical proposition of cultural competence development could be used not only to guide initial and continuing nursing education, but also to help redefine quality of care in a culturally diverse context. Copyright © 2015 Elsevier Ltd. All rights reserved.

  14. Sub-micron opto-chemical probes for studying living neurons

    NASA Astrophysics Data System (ADS)

    Hossein-Zadeh, M.; Delgado, J.; Schweizer, F.; Lieberman, R.

    2017-02-01

    We have fabricated sub-micron opto-chemical probes for pH, oxygen and calcium monitoring and demonstrated their application in intracellular and extracellular monitoring of neurons (cortical neuronal cultures and acute hippocampal slices). Using these probes, we have measured extracellular pH in the stratum radiatum of the CA1 region of mouse hippocampus upon stimulation of presynaptic Schaffer collateral axons. Synaptic transmission was monitored using standard electrophysiological techniques. We find that the local pH transiently changes in response to synaptic stimulation. In addition, the geometry of the functionalized region on the probe combined with high sensitivity imaging enables simultaneous monitoring of spatially adjacent but distinct compartments. As proof of concept we impaled cultured neurons with the probe measured calcium and pH inside as well as directly outside of neurons as we changed the pH and calcium concentration in the physiological solution in the perfusion chamber. As such these probes can be used to study the impact of the environment on both cellular and extra-cellular space. Additionally as the chemical properties of the surrounding medium can be controlled and monitored with high precision, these probes enable differential measurement of the target parameter referenced to a stable bath. This approach eliminates the uncertainties associated with non-chemical fluctuations in the fluorescent emission and result in a self-calibrated opto-chemical probe. We have also demonstrated multifunctional probes that are capable of measuring up to three parameters in the extracellular space in brain slices.

  15. Bis-enoxacin Inhibits Bone Resorption and Orthodontic Tooth Movement

    PubMed Central

    Toro, E.J.; Zuo, J.; Guiterrez, A.; La Rosa, R.L.; Gawron, A.J.; Bradaschia-Correa, V.; Arana-Chavez, V.; Dolce, C.; Rivera, M.F.; Kesavalu, L.; Bhattacharyya, I.; Neubert, J.K.; Holliday, L.S.

    2013-01-01

    Enoxacin inhibits binding between the B-subunit of vacuolar H+-ATPase (V-ATPase) and microfilaments, and also between osteoclast formation and bone resorption in vitro. We hypothesized that a bisphosphonate derivative of enoxacin, bis-enoxacin (BE), which was previously studied as a bone-directed antibiotic, might have similar activities. BE shared a number of characteristics with enoxacin: It blocked binding between the recombinant B-subunit and microfilaments and inhibited osteoclastogenesis in cell culture with IC50s of about 10 µM in each case. BE did not alter the relative expression levels of various osteoclast-specific proteins. Even though tartrate-resistant acid phosphatase 5b was expressed, proteolytic activation of the latent pro-enzyme was inhibited. However, unlike enoxacin, BE stimulated caspase-3 activity. BE bound to bone slices and inhibited bone resorption by osteoclasts on BE-coated bone slices in cell culture. BE reduced the amount of orthodontic tooth movement achieved in rats after 28 days. Analysis of these data suggests that BE is a novel anti-resorptive molecule that is active both in vitro and in vivo and may have clinical uses. Abbreviations: BE, bis-enoxacin; V-ATPase, vacuolar H+-ATPase; TRAP, tartrate-resistant acid phosphatase; αMEM D10, minimal essential media, alpha modification with 10% fetal bovine serum; SDS-PAGE, sodium dodecyl sulfate-polyacrylamide gel electrophoresis; RANKL, receptor activator of nuclear factor kappa B-ligand; NFATc1, nuclear factor of activated T-cells; ADAM, a disintegrin and metalloprotease domain; OTM, orthodontic tooth movement. PMID:23958763

  16. Lipopolysaccharide-induced dopaminergic cell death in rat midbrain slice cultures: role of inducible nitric oxide synthase and protection by indomethacin.

    PubMed

    Shibata, Haruki; Katsuki, Hiroshi; Nishiwaki, Mayumi; Kume, Toshiaki; Kaneko, Shuji; Akaike, Akinori

    2003-09-01

    Glial cell activation associated with inflammatory reaction may contribute to pathogenic processes of neurodegenerative disorders, through production of several cytotoxic molecules. We investigated the consequences of glial activation by interferon-gamma (IFN-gamma)/lipopolysaccharide (LPS) in rat midbrain slice cultures. Application of IFN-gamma followed by LPS caused dopaminergic cell death and accompanying increases in nitrite production and lactate dehydrogenase release. Aminoguanidine, an inhibitor of inducible nitric oxide synthase (iNOS), or SB203580, an inhibitor of p38 mitogen-activated protein kinase, prevented dopaminergic cell loss as well as nitrite production. SB203580 also suppressed expression of iNOS and cyclooxygenase-2 (COX-2) induced by IFN-gamma/LPS. A COX inhibitor indomethacin protected dopaminergic neurons from IFN-gamma/LPS-induced injury, whereas selective COX-2 inhibitors such as NS-398 and nimesulide did not. Notably, indomethacin was able to attenuate neurotoxicity of a nitric oxide (NO) donor. Neutralizing antibodies against tumour necrosis factor-alpha and interleukin-1beta did not inhibit dopaminergic cell death caused by IFN-gamma/LPS, although combined application of these antibodies blocked lactate dehydrogenase release and decrease in the number of non-dopaminergic neurons. These results indicate that iNOS-derived NO plays a crucial role in IFN-gamma/LPS-induced dopaminergic cell death, and that indomethacin exerts protective effect by mechanisms probably related to NO neurotoxicity rather than through COX inhibition.

  17. Extracellular caspase-6 drives murine inflammatory pain via microglial TNF-α secretion

    PubMed Central

    Berta, Temugin; Park, Chul-Kyu; Xu, Zhen-Zhong; Xie, Ruo-Gang; Liu, Tong; Lü, Ning; Liu, Yen-Chin; Ji, Ru-Rong

    2014-01-01

    Increasing evidence indicates that the pathogenesis of neuropathic pain is mediated through spinal cord microglia activation. The intracellular protease caspase-6 (CASP6) is known to regulate neuronal apoptosis and axonal degeneration; however, the contribution of microglia and CASP6 in modulating synaptic transmission and pain is unclear. Here, we found that CASP6 is expressed specifically in C-fiber axonal terminals in the superficial spinal cord dorsal horn. Animals exposed to intraplantar formalin or bradykinin injection exhibited CASP6 activation in the dorsal horn. Casp6-null mice had normal baseline pain, but impaired inflammatory pain responses. Furthermore, formalin-induced second-phase pain was suppressed by spinal injection of CASP6 inhibitor or CASP6-neutralizing antibody, as well as perisciatic nerve injection of CASP6 siRNA. Recombinant CASP6 (rCASP6) induced marked TNF-α release in microglial cultures, and most microglia within the spinal cord expressed Tnfa. Spinal injection of rCASP6 elicited TNF-α production and microglia-dependent pain hypersensitivity. Evaluation of excitatory postsynaptic currents (EPSCs) revealed that rCASP6 rapidly increased synaptic transmission in spinal cord slices via TNF-α release. Interestingly, the microglial inhibitor minocycline suppressed rCASP6 but not TNF-α–induced synaptic potentiation. Finally, rCASP6-activated microglial culture medium increased EPSCs in spinal cord slices via TNF-α. Together, these data suggest that CASP6 released from axonal terminals regulates microglial TNF-α secretion, synaptic plasticity, and inflammatory pain. PMID:24531553

  18. Family Life and Developmental Idealism in Yazd, Iran

    PubMed Central

    Abbasi-Shavazi, Mohammad Jalal; Askari-Nodoushan, Abbas

    2012-01-01

    BACKGROUND This paper is motivated by the theory that developmental idealism has been disseminated globally and has become an international force for family and demographic change. Developmental idealism is a set of cultural beliefs and values about development and how development relates to family and demographic behavior. It holds that modern societies are causal forces producing modern families, that modern families help to produce modern societies, and that modern family change is to be expected. OBJECTIVE We examine the extent to which developmental idealism has been disseminated in Iran. We also investigate predictors of the dissemination of developmental idealism. METHODS We use survey data collected in 2007 from a sample of women in Yazd, a city in Iran. We examine the distribution of developmental idealism in the sample and the multivariate predictors of developmental idealism. RESULTS We find considerable support for the expectation that many elements of developmental idealism have been widely disseminated. Statistically significant majorities associate development with particular family attributes, believe that development causes change in families, believe that fertility reductions and age-at-marriage increases help foster development, and perceive family trends in Iran headed toward modernity. As predicted, parental education, respondent education, and income affect adherence to developmental idealism. CONCLUSIONS Developmental idealism has been widely disseminated in Yazd, Iran and is related to social and demographic factors in predicted ways. COMMENTS Although our data come from only one city, we expect that developmental idealism has been widely distributed in Iran, with important implications for family and demographic behavior. PMID:22942772

  19. Murine precision-cut liver slices (PCLS): a new tool for studying tumor microenvironments and cell signaling ex vivo.

    PubMed

    Koch, Alexandra; Saran, Shashank; Tran, Doan Duy Hai; Klebba-Färber, Sabine; Thiesler, Hauke; Sewald, Katherina; Schindler, Susann; Braun, Armin; Klopfleisch, Robert; Tamura, Teruko

    2014-11-07

    One of the most insidious characteristics of cancer is its spread to and ability to compromise distant organs via the complex process of metastasis. Communication between cancer cells and organ-resident cells via cytokines/chemokines and direct cell-cell contacts are key steps for survival, proliferation and invasion of metastasized cancer cells in organs. Precision-cut liver slices (PCLS) are considered to closely reflect the in vivo situation and are potentially useful for studying the interaction of cancer cells with liver-resident cells as well as being a potentially useful tool for screening anti-cancer reagents. Application of the PCLS technique in the field of cancer research however, has not yet been well developed. We established the mouse PCLS system using perfluorodecalin (PFD) as an artificial oxygen carrier. Using this system we show that the adherence of green fluorescent protein (GFP) labeled MDA-MB-231 (highly invasive) cells to liver tissue in the PCLS was 5-fold greater than that of SK-BR-3 (less invasive) cells. In addition, we generated PCLS from THOC5, a member of transcription/export complex (TREX), knockout (KO) mice. The PCLS still expressed Gapdh or Albumin mRNAs at normal levels, while several chemokine/growth factor or metalloprotease genes, such as Cxcl12, Pdgfa, Tgfb, Wnt11, and Mmp1a genes were downregulated more than 2-fold. Interestingly, adhesion of cancer cells to THOC5 KO liver slices was far less (greater than 80% reduction) than to wild-type liver slices. Mouse PCLS cultures in the presence of PFD may serve as a useful tool for screening local adherence and invasiveness of individual cancer cells, since single cells can be observed. This method may also prove useful for identification of genes in liver-resident cells that support cancer invasion by using PCLS from transgenic liver.

  20. The topology of large-scale structure. VI - Slices of the universe

    NASA Astrophysics Data System (ADS)

    Park, Changbom; Gott, J. R., III; Melott, Adrian L.; Karachentsev, I. D.

    1992-03-01

    Results of an investigation of the topology of large-scale structure in two observed slices of the universe are presented. Both slices pass through the Coma cluster and their depths are 100 and 230/h Mpc. The present topology study shows that the largest void in the CfA slice is divided into two smaller voids by a statistically significant line of galaxies. The topology of toy models like the white noise and bubble models is shown to be inconsistent with that of the observed slices. A large N-body simulation was made of the biased cloud dark matter model and the slices are simulated by matching them in selection functions and boundary conditions. The genus curves for these simulated slices are spongelike and have a small shift in the direction of a meatball topology like those of observed slices.

  1. The topology of large-scale structure. VI - Slices of the universe

    NASA Technical Reports Server (NTRS)

    Park, Changbom; Gott, J. R., III; Melott, Adrian L.; Karachentsev, I. D.

    1992-01-01

    Results of an investigation of the topology of large-scale structure in two observed slices of the universe are presented. Both slices pass through the Coma cluster and their depths are 100 and 230/h Mpc. The present topology study shows that the largest void in the CfA slice is divided into two smaller voids by a statistically significant line of galaxies. The topology of toy models like the white noise and bubble models is shown to be inconsistent with that of the observed slices. A large N-body simulation was made of the biased cloud dark matter model and the slices are simulated by matching them in selection functions and boundary conditions. The genus curves for these simulated slices are spongelike and have a small shift in the direction of a meatball topology like those of observed slices.

  2. Comparison of 640-Slice Multidetector Computed Tomography Versus 32-Slice MDCT for Imaging of the Osteo-odonto-keratoprosthesis Lamina.

    PubMed

    Norris, Joseph M; Kishikova, Lyudmila; Avadhanam, Venkata S; Koumellis, Panos; Francis, Ian S; Liu, Christopher S C

    2015-08-01

    To investigate the efficacy of 640-slice multidetector computed tomography (MDCT) for detecting osteo-odonto laminar resorption in the osteo-odonto-keratoprosthesis (OOKP) compared with the current standard 32-slice MDCT. Explanted OOKP laminae and bone-dentine fragments were scanned using 640-slice MDCT (Aquilion ONE; Toshiba) and 32-slice MDCT (LightSpeed Pro32; GE Healthcare). Pertinent comparisons including image quality, radiation dose, and scanning parameters were made. Benefits of 640-slice MDCT over 32-slice MDCT were shown. Key comparisons of 640-slice MDCT versus 32-slice MDCT included the following: percentage difference and correlation coefficient between radiological and anatomical measurements, 1.35% versus 3.67% and 0.9961 versus 0.9882, respectively; dose-length product, 63.50 versus 70.26; rotation time, 0.175 seconds versus 1.000 seconds; and detector coverage width, 16 cm versus 2 cm. Resorption of the osteo-odonto lamina after OOKP surgery can result in potentially sight-threatening complications, hence it warrants regular monitoring and timely intervention. MDCT remains the gold standard for radiological assessment of laminar resorption, which facilitates detection of subtle laminar changes earlier than the onset of clinical signs, thus indicating when preemptive measures can be taken. The 640-slice MDCT exhibits several advantages over traditional 32-slice MDCT. However, such benefits may not offset cost implications, except in rare cases, such as in young patients who might undergo years of radiation exposure.

  3. Age-specific function of α5β1 integrin in microglial migration during early colonization of the developing mouse cortex.

    PubMed

    Smolders, Sophie Marie-Thérèse; Swinnen, Nina; Kessels, Sofie; Arnauts, Kaline; Smolders, Silke; Le Bras, Barbara; Rigo, Jean-Michel; Legendre, Pascal; Brône, Bert

    2017-07-01

    Microglia, the immune cells of the central nervous system, take part in brain development and homeostasis. They derive from primitive myeloid progenitors that originate in the yolk sac and colonize the brain mainly through intensive migration. During development, microglial migration speed declines which suggests that their interaction with the microenvironment changes. However, the matrix-cell interactions allowing dispersion within the parenchyma are unknown. Therefore, we aimed to better characterize the migration behavior and to assess the role of matrix-integrin interactions during microglial migration in the embryonic brain ex vivo. We focused on microglia-fibronectin interactions mediated through the fibronectin receptor α5β1 integrin because in vitro work indirectly suggested a role for this ligand-receptor pair. Using 2-photon time-lapse microscopy on acute ex vivo embryonic brain slices, we found that migration occurs in a saltatory pattern and is developmentally regulated. Most importantly, there is an age-specific function of the α5β1 integrin during microglial cortex colonization. At embryonic day (E) 13.5, α5β1 facilitates migration while from E15.5, it inhibits migration. These results indicate a developmentally regulated function of α5β1 integrin in microglial migration during colonization of the embryonic brain. © 2017 Wiley Periodicals, Inc.

  4. Longitudinal and Integrative Tests of Family Stress Model Effects on Mexican-Origin Adolescents

    PubMed Central

    White, Rebecca M. B.; Liu, Yu; Nair, Rajni L.; Tein, Jenn-Yun

    2015-01-01

    The family stress model represents a common framework through which to examine the effects of environmental stressors on adolescent adjustment. The model suggests that economic and neighborhood stressors influence youth adjustment via disruptions to parenting. Incorporating integrative developmental theory, we examined the degree to which parents’ cultural value orientations mitigated the effects of stressors on parenting disruptions and the degree to which environmental adversity qualified the effect of parenting on adolescent adjustment. We tested the hypothesized Integrative Family Stress Model longitudinally in a sample of mother-youth dyads (N = 749) and father-youth dyads (N = 467) from Mexican origin families, across three times points spanning early to middle adolescence. Providing the first longitudinal evidence of family stress mediated effects, mothers’ perceptions of economic pressure were associated with increases in adolescent externalizing symptoms five years later via intermediate increases in harsh parenting. The remaining findings supported the notion that integrative developmental theory can inform family stress model hypothesis testing that is culturally and contextually relevant for wide range of diverse families and youth. For example, fathers’ perceptions of economic pressure and neighborhood danger had important implications for adolescent internalizing, via reductions in paternal warmth, but only at certain levels of neighborhood adversity. Mothers’ familism value orientations mitigated the effects of economic pressure on maternal warmth, protecting their adolescents from experiencing developmental costs associated with environmental stressors. Results are discussed in terms of identifying how integrative developmental theory intersects with the family stress model to set diverse youth on different developmental pathways. PMID:25751100

  5. Longitudinal and integrative tests of family stress model effects on Mexican origin adolescents.

    PubMed

    White, Rebecca M B; Liu, Yu; Nair, Rajni L; Tein, Jenn-Yun

    2015-05-01

    The family stress model represents a common framework through which to examine the effects of environmental stressors on adolescent adjustment. The model suggests that economic and neighborhood stressors influence youth adjustment via disruptions to parenting. Incorporating integrative developmental theory, we examined the degree to which parents' cultural value orientations mitigated the effects of stressors on parenting disruptions and the degree to which environmental adversity qualified the effect of parenting on adolescent adjustment. We tested the hypothesized integrative family stress model longitudinally in a sample of mother-youth dyads (N = 749) and father-youth dyads (N = 467) from Mexican origin families, across 3 times points spanning early to middle adolescence. Providing the first longitudinal evidence of family stress mediated effects, mothers' perceptions of economic pressure were associated with increases in adolescent externalizing symptoms 5 years later via intermediate increases in harsh parenting. The remaining findings supported the notion that integrative developmental theory can inform family stress model hypothesis testing that is culturally and contextually relevant for a wide range of diverse families and youth. For example, fathers' perceptions of economic pressure and neighborhood danger had important implications for adolescent internalizing, via reductions in paternal warmth, but only at certain levels of neighborhood adversity. Mothers' familism value orientations mitigated the effects of economic pressure on maternal warmth, protecting their adolescents from experiencing developmental costs associated with environmental stressors. Results are discussed in terms of identifying how integrative developmental theory intersects with the family stress model to set diverse youth on different developmental pathways. (c) 2015 APA, all rights reserved).

  6. Slicing of silicon into sheet material: Silicon sheet growth development for the large area silicon sheet task of the low cost silicon solar array project

    NASA Technical Reports Server (NTRS)

    Holden, S. C.

    1976-01-01

    Multiblade slurry sawing is used to slice 10 cm diameter silicon ingots into wafers 0.024 cm thick using 0.050 cm of silicon per slice (0.026 cm kerf loss). Total slicing time is less than twenty hours, and 143 slices are produced simultaneously. Productivity (slice area per hour per blade) is shown as a function or blade load and thickness, and abrasive size. Finer abrasive slurries cause a reduction in slice productivity, and thin blades cause a reduction of wafer accuracy. Sawing induced surface damage is found to extend 18 microns into the wafer.

  7. Precision-cut liver slices as a model for the early onset of liver fibrosis to test antifibrotic drugs

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

    Westra, Inge M.; Oosterhuis, Dorenda; Groothuis, Geny M.M.

    Induction of fibrosis during prolonged culture of precision-cut liver slices (PCLS) was reported. In this study, the use of rat PCLS was investigated to further characterize the mechanism of early onset of fibrosis in this model and the effects of antifibrotic compounds. Rat PCLS were incubated for 48 h, viability was assessed by ATP and gene expression of PDGF-B and TGF-β1 and the fibrosis markers Hsp47, αSma and Pcol1A1 and collagen1 protein expressions were determined. The effects of the antifibrotic drugs imatinib, sorafenib and sunitinib, PDGF-pathway inhibitors, and perindopril, valproic acid, rosmarinic acid, tetrandrine and pirfenidone, TGFβ-pathway inhibitors, were determined.more » After 48 h of incubation, viability of the PCLS was maintained and gene expression of PDGF-B was increased while TGF-β1 was not changed. Hsp47, αSma and Pcol1A1 gene expressions were significantly elevated in PCLS after 48 h, which was further increased by PDGF-BB and TGF-β1. The increased gene expression of fibrosis markers was inhibited by all three PDGF-inhibitors, while TGFβ-inhibitors showed marginal effects. The protein expression of collagen 1 was inhibited by imatinib, perindopril, tetrandrine and pirfenidone. In conclusion, the increased gene expression of PDGF-B and the down-regulation of fibrosis markers by PDGF-pathway inhibitors, together with the absence of elevated TGF-β1 gene expression and the limited effect of the TGFβ-pathway inhibitors, indicated the predominance of the PDGF pathway in the early onset of fibrosis in PCLS. PCLS appear a useful model for research of the early onset of fibrosis and for testing of antifibrotic drugs acting on the PDGF pathway. - Highlights: • During culture, fibrosis markers increased in precision-cut liver slices (PCLS). • Gene expression of PDGF-β was increased, while TGFβ was not changed in rat PCLS. • PDGF-pathway inhibitors down-regulated this increase of fibrosis markers. • TGFβ-pathway inhibitors had only minor effects on fibrosis markers. • Rat PCLS can be used to study the early onset of fibrosis.« less

  8. Thin slices of child personality: Perceptual, situational, and behavioral contributions.

    PubMed

    Tackett, Jennifer L; Herzhoff, Kathrin; Kushner, Shauna C; Rule, Nicholas

    2016-01-01

    The present study examined whether thin-slice ratings of child personality serve as a resource-efficient and theoretically valid measurement of child personality traits. We extended theoretical work on the observability, perceptual accuracy, and situational consistency of childhood personality traits by examining intersource and interjudge agreement, cross-situational consistency, and convergent, divergent, and predictive validity of thin-slice ratings. Forty-five unacquainted independent coders rated 326 children's (ages 8-12) personality in 1 of 15 thin-slice behavioral scenarios (i.e., 3 raters per slice, for over 14,000 independent thin-slice ratings). Mothers, fathers, and children rated children's personality, psychopathology, and competence. We found robust evidence for correlations between thin-slice and mother/father ratings of child personality, within- and across-task consistency of thin-slice ratings, and convergent and divergent validity with psychopathology and competence. Surprisingly, thin-slice ratings were more consistent across situations in this child sample than previously found for adults. Taken together, these results suggest that thin slices are a valid and reliable measure to assess child personality, offering a useful method of measurement beyond questionnaires, helping to address novel questions of personality perception and consistency in childhood. (c) 2016 APA, all rights reserved).

  9. High-order multiband encoding in the heart.

    PubMed

    Cunningham, Charles H; Wright, Graham A; Wood, Michael L

    2002-10-01

    Spatial encoding with multiband selective excitation (e.g., Hadamard encoding) has been restricted to a small number of slices because the RF pulse becomes unacceptably long when more than about eight slices are encoded. In this work, techniques to shorten multiband RF pulses, and thus allow larger numbers of slices, are investigated. A method for applying the techniques while retaining the capability of adaptive slice thickness is outlined. A tradeoff between slice thickness and pulse duration is shown. Simulations and experiments with the shortened pulses confirmed that motion-induced excitation profile blurring and phase accrual were reduced. The connection between gradient hardware limitations, slice thickness, and flow sensitivity is shown. Excitation profiles for encoding 32 contiguous slices of 1-mm thickness were measured experimentally, and the artifact resulting from errors in timing of RF pulse relative to gradient was investigated. A multiband technique for imaging 32 contiguous 2-mm slices, with adaptive slice thickness, was developed and demonstrated for coronary artery imaging in healthy subjects. With the ability to image high numbers of contiguous slices, using relatively short (1-2 ms) RF pulses, multiband encoding has been advanced further toward practical application. Copyright 2002 Wiley-Liss, Inc.

  10. Comparative Analysis of Human and Rodent Brain Primary Neuronal Culture Spontaneous Activity Using Micro-Electrode Array Technology.

    PubMed

    Napoli, Alessandro; Obeid, Iyad

    2016-03-01

    Electrical activity in embryonic brain tissue has typically been studied using Micro Electrode Array (MEA) technology to make dozens of simultaneous recordings from dissociated neuronal cultures, brain stem cell progenitors, or brain slices from fetal rodents. Although these rodent neuronal primary culture electrical properties are mostly investigated, it has not been yet established to what extent the electrical characteristics of rodent brain neuronal cultures can be generalized to those of humans. A direct comparison of spontaneous spiking activity between rodent and human primary neurons grown under the same in vitro conditions using MEA technology has never been carried out before and will be described in the present study. Human and rodent dissociated fetal brain neuronal cultures were established in-vitro by culturing on a glass grid of 60 planar microelectrodes neurons under identical conditions. Three different cultures of human neurons were produced from tissue sourced from a single aborted fetus (at 16-18 gestational weeks) and these were compared with seven different cultures of embryonic rat neurons (at 18 gestational days) originally isolated from a single rat. The results show that the human and rodent cultures behaved significantly differently. Whereas the rodent cultures demonstrated robust spontaneous activation and network activity after only 10 days, the human cultures required nearly 40 days to achieve a substantially weaker level of electrical function. These results suggest that rat neuron preparations may yield inferences that do not necessarily transfer to humans. © 2015 Wiley Periodicals, Inc.

  11. Child and Adolescent Use of Mobile Phones: An Unparalleled Complex Developmental Phenomenon.

    PubMed

    Yan, Zheng

    2018-01-01

    This article addresses why children's use of mobile phones is an unparalleled complex developmental phenomenon in hopes of providing a broad context for this special section. It first outlines mobile phones as a sophisticated personalized and multifunction technology. Then it presents mobile phone use by children as an unparalleled complex developmental phenomenon on the basis of its four behavioral elements, two mobile cultures, and two developmental processes. It further illustrates the existing knowledge about children's mobile phones use that has been accumulated over the past 23 years and highlights 12 most studied topics, especially distracted driving and radiation exposure. It concludes with three types of scientific contributions made by the 12 articles in the special section. © 2017 The Author. Child Development © 2017 Society for Research in Child Development, Inc.

  12. RETROSPECTIVE DETECTION OF INTERLEAVED SLICE ACQUISITION PARAMETERS FROM FMRI DATA

    PubMed Central

    Parker, David; Rotival, Georges; Laine, Andrew; Razlighi, Qolamreza R.

    2015-01-01

    To minimize slice excitation leakage to adjacent slices, interleaved slice acquisition is nowadays performed regularly in fMRI scanners. In interleaved slice acquisition, the number of slices skipped between two consecutive slice acquisitions is often referred to as the ‘interleave parameter’; the loss of this parameter can be catastrophic for the analysis of fMRI data. In this article we present a method to retrospectively detect the interleave parameter and the axis in which it is applied. Our method relies on the smoothness of the temporal-distance correlation function, which becomes disrupted along the axis on which interleaved slice acquisition is applied. We examined this method on simulated and real data in the presence of fMRI artifacts such as physiological noise, motion, etc. We also examined the reliability of this method in detecting different types of interleave parameters and demonstrated an accuracy of about 94% in more than 1000 real fMRI scans. PMID:26161244

  13. Pre-Service Teachers and Study Abroad: A Reflective, Experiential Sojourn to Increase Intercultural Competence and Translate the Experience into Culturally Relevant Pedagogy

    ERIC Educational Resources Information Center

    Roller, Kathleen Marie

    2012-01-01

    This study used a randomized experimental, mixed methods approach to examine whether a stand-apart course curriculum based on experiential learning theory, Developmental Model of Intercultural Sensitivity (DMIS) theory (Bennett, 1993) and culturally relevant pedagogy influenced students' intercultural competence within the context of a study…

  14. Assessing Autistic Traits in a Taiwan Preschool Population: Cross-Cultural Validation of the Social Responsiveness Scale (SRS)

    ERIC Educational Resources Information Center

    Wang, Jessica; Lee, Li-Ching; Chen, Ying-Sheue; Hsu, Ju-Wei

    2012-01-01

    The cross-cultural validity of the Mandarin-adaptation of the social responsiveness scale (SRS) was examined in a sample of N = 307 participants in Taiwan, 140 typically developing and 167 with clinically-diagnosed developmental disorders. This scale is an autism assessment tool that provides a quantitative rather than categorical measure of…

  15. Virtual Journey Coupled with Face-to-Face Exchange: Enhancing the Cultural Sensitivity and Competence of Graduate Students

    ERIC Educational Resources Information Center

    Ellenwood, Audrey E.; Snyders, Frederik J. A.

    2010-01-01

    This study attempted to enhance cultural sensitivity for graduate students at an American and a South African university, using a six-week online List Serv, email buddy exchange, and two-week face-to-face experience. After the course was over, results of the intercultural developmental inventory, using "t"-tests for related samples,…

  16. Adapting a Developmental Screening Measure: Exploring the Effects of Language and Culture on a Parent-Completed SocialEmotional Screening Test

    ERIC Educational Resources Information Center

    Chen, Chieh-Yu; Chen, Ching-I; Squires, Jane; Bian, Xiaoyan; Heo, Kay H.; Filgueiras, Alberto; Kalinina, Svetlana; Samarina, Larissa; Ermolaeva, Evgeniya; Xie, Huichao; Yu, Ting-Ying; Wu, Pei-Fang; Landeira-Fernandez, Jesus

    2017-01-01

    Ages & Stages Questionnaires: Social-Emotional (ASQ:SE) is a widely used screening instrument for detecting social-emotional difficulties in infants and young children. To use a screening instrument across cultures and countries, it is necessary to identify potential item-level biases and ensure item equivalence. This study investigated the…

  17. Identity Development and Future Orientation in Immigrant Adolescents and Young Adults: A Narrative View of Cultural Transitions From Ethiopia to Israel.

    PubMed

    Flum, Hanoch; Buzukashvili, Tamara

    2018-06-01

    This paper examines a major aspect of identity development in the context of cultural transition. Following Eriksonian psychosocial and sociocultural perspectives, it investigates self-continuity and identity integration in light of inherent discontinuity among young immigrants. More specifically, this examination draws on three distinct narrative studies, within the framework of Dynamic Narrative Approach, with first- and second-generation adolescents and young adult Ethiopian immigrants to Israel. Their negotiations of identity, with a focus on their narrative construction of past, present, and future across life domains (education, career, military service, family), are illustrated in this article in a variety of developmental paths. Dynamics of reciprocity between early life experiences and future orientation are revealed in the narratives. A capacity to connect cultural resources in the past with challenges in the new culture is identified as a key. By processing them and bringing them up-to-date, meaning becomes relevant to current experiences and developmental challenges. Across the three distinct studies, a variety of exploratory activities and relational qualities are found to facilitate or impede the reconstruction and integration of identity. © 2018 Wiley Periodicals, Inc.

  18. Developmental potential of murine germinal vesicle stage cumulus-oocyte complexes following exposure to dimethylsulphoxide or cryopreservation: loss of membrane integrity of cumulus cells after thawing.

    PubMed

    Ruppert-Lingham, C J; Paynter, S J; Godfrey, J; Fuller, B J; Shaw, R W

    2003-02-01

    Cumulus cells of the cumulus-oocyte complex (COC) are important in oocyte maturation. Thus, in preserving immature oocytes it is prudent to also preserve their associated cumulus cells. The survival and function of oocytes and their associated cumulus cells was assessed following cryopreservation or exposure to cryoprotectant without freezing. Immature COCs were collected from mice primed with pregnant mare's serum. COCs were either slow-cooled or exposed to 1.5 mol/l dimethylsulphoxide without freezing. Treated and fresh COCs were stained for membrane integrity or, after in-vitro maturation and IVF, were assessed for developmental capability. Development of cumulus-denuded fresh oocytes, as well as denuded and frozen-thawed oocytes co-cultured with fresh cumulus cells, was assessed. Slow-cooled oocytes had significantly reduced coverage by intact cumulus cells compared with fresh COCs. Cumulus cell association and developmental capability were not substantially affected by exposure to cryoprotectant without freezing. Denuded fresh oocytes and cryopreserved COCs had decreased developmental potential that was not overcome by co-culture with fresh cumulus cells. Loss of association between oocyte and cumulus cells was induced by cryopreservation, but not by treatment with cryoprotectant alone. The data indicate that direct physical contact between cumulus cells and the oocyte, throughout maturation, improves subsequent embryo development.

  19. Can multi-slice or navigator-gated R2* MRI replace single-slice breath-hold acquisition for hepatic iron quantification?

    PubMed

    Loeffler, Ralf B; McCarville, M Beth; Wagstaff, Anne W; Smeltzer, Matthew P; Krafft, Axel J; Song, Ruitian; Hankins, Jane S; Hillenbrand, Claudia M

    2017-01-01

    Liver R2* values calculated from multi-gradient echo (mGRE) magnetic resonance images (MRI) are strongly correlated with hepatic iron concentration (HIC) as shown in several independently derived biopsy calibration studies. These calibrations were established for axial single-slice breath-hold imaging at the location of the portal vein. Scanning in multi-slice mode makes the exam more efficient, since whole-liver coverage can be achieved with two breath-holds and the optimal slice can be selected afterward. Navigator echoes remove the need for breath-holds and allow use in sedated patients. To evaluate if the existing biopsy calibrations can be applied to multi-slice and navigator-controlled mGRE imaging in children with hepatic iron overload, by testing if there is a bias-free correlation between single-slice R2* and multi-slice or multi-slice navigator controlled R2*. This study included MRI data from 71 patients with transfusional iron overload, who received an MRI exam to estimate HIC using gradient echo sequences. Patient scans contained 2 or 3 of the following imaging methods used for analysis: single-slice images (n = 71), multi-slice images (n = 69) and navigator-controlled images (n = 17). Small and large blood corrected region of interests were selected on axial images of the liver to obtain R2* values for all data sets. Bland-Altman and linear regression analysis were used to compare R2* values from single-slice images to those of multi-slice images and navigator-controlled images. Bland-Altman analysis showed that all imaging method comparisons were strongly associated with each other and had high correlation coefficients (0.98 ≤ r ≤ 1.00) with P-values ≤0.0001. Linear regression yielded slopes that were close to 1. We found that navigator-gated or breath-held multi-slice R2* MRI for HIC determination measures R2* values comparable to the biopsy-validated single-slice, single breath-hold scan. We conclude that these three R2* methods can be interchangeably used in existing R2*-HIC calibrations.

  20. Cultural relativism: maintenance of genomic imprints in pluripotent stem cell culture systems.

    PubMed

    Greenberg, Maxim Vc; Bourc'his, Déborah

    2015-04-01

    Pluripotent stem cells (PSCs) in culture have become a widely used model for studying events occurring during mammalian development; they also present an exciting avenue for therapeutics. However, compared to their in vivo counterparts, cultured PSC derivatives have unique properties, and it is well established that their epigenome is sensitive to medium composition. Here we review the specific effects on genomic imprints in various PSC types and culture systems. Imprinted gene regulation is developmentally important, and imprinting defects have been associated with several human diseases. Therefore, imprint abnormalities in PSCs may have considerable consequences for downstream applications. Copyright © 2015 Elsevier Ltd. All rights reserved.

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