Sample records for conditioned medium cm

  1. Prospect of stem cell conditioned medium in regenerative medicine.

    PubMed

    Pawitan, Jeanne Adiwinata

    2014-01-01

    Stem cell-derived conditioned medium has a promising prospect to be produced as pharmaceuticals for regenerative medicine. To investigate various methods to obtain stem cell-derived conditioned medium (CM) to get an insight into their prospect of application in various diseases. Systematic review using keywords "stem cell" and "conditioned medium" or "secretome" and "therapy." Data concerning treated conditions/diseases, type of cell that was cultured, medium and supplements to culture the cells, culture condition, CM processing, growth factors and other secretions that were analyzed, method of application, and outcome were noted, grouped, tabulated, and analyzed. Most of CM using studies showed good results. However, the various CM, even when they were derived from the same kind of cells, were produced by different condition, that is, from different passage, culture medium, and culture condition. The growth factor yields of the various types of cells were available in some studies, and the cell number that was needed to produce CM for one application could be computed. Various stem cell-derived conditioned media were tested on various diseases and mostly showed good results. However, standardized methods of production and validations of their use need to be conducted.

  2. Conditioned Medium from Placental Mesenchymal Stem Cells Reduces Oxidative Stress during the Cryopreservation of Ex Vivo Expanded Umbilical Cord Blood Cells.

    PubMed

    Kadekar, Darshana; Rangole, Sonal; Kale, Vaijayanti; Limaye, Lalita

    2016-01-01

    The limited cell dose in umbilical cord blood (UCB) necessitates ex vivo expansion of UCB. Further, the effective cryopreservation of these expanded cells is important in widening their use in the clinics. During cryopreservation, cells experience oxidative stress due to the generation of reactive oxygen species (ROS). Conditioned medium from mesenchymal stem cells (MSCs-CM) has been shown to alleviate the oxidative stress during wound healing, Alzheimer's disease and ischemic disease. This premise prompted us to investigate the influence of MSCs-CM during cryopreservation of expanded UCB cells. CM-was collected from cord/placental MSCs(C-MSCs-CM, P-MSC-CM). UCB CD34+cells were expanded as suspension cultures in serum free medium containing cytokines for 10 days. Cells were frozen with/without C-MSCs-CM and or P-MSCs-CM in the conventional freezing medium containing 20%FCS +10%DMSO using a programmable freezer and stored in liquid nitrogen. Upon revival, cells frozen with MSCs-CM were found to be superior to cells frozen in conventional medium in terms of viability, CD34+content and clonogenecity. Priming of revived cells for 48 hrs with MSCs-CM further improved their transplantation ability, as compared to those cultured without MSCs-CM. P-MSCs-CM radically reduced the oxidative stress in cryopreserved cells, resulting in better post thaw functionality of CD34+ cells than with C-MSCs-CM. The observed cryoprotective effect of MSCs-CM was primarily due to anti-oxidative and anti-apoptotic properties of the MSCs-CM and not because of the exosomes secreted by them. Our data suggest that MSCs-CM can serve as a valuable additive to the freezing or the priming medium for expanded UCB cells, which would increase their clinical applicability.

  3. Conditioned medium of adipose-derived stromal cell culture in three-dimensional bioreactors for enhanced wound healing.

    PubMed

    Kwon, Sun Hyun; Bhang, Suk Ho; Jang, Hyeon-Ki; Rhim, Taiyoun; Kim, Byung-Soo

    2015-03-01

    It was previously shown that human adipose-derived stromal cell (hADSC)-conditioned medium (CM) promotes wound healing. An essential part of the wound healing process is neovascularization in the wound bed. We hypothesized that CM prepared from hADSCs cultured as spheroids in three-dimensional suspension bioreactors (spheroid CM) would contain much higher concentrations of angiogenic growth factors secreted by hADSCs, induce a higher extent of neovascularization in the wound bed, and improve wound healing as compared with CM prepared by conventional monolayer culture (monolayer CM). The concentrations of angiogenic growth factors (i.e., vascular endothelial growth factor, basic fibroblast growth factor, and hepatocyte growth factor) in spheroid CM were 20- to 145-fold higher than those in monolayer CM. Either fresh medium, monolayer CM, or spheroid CM was administered to full-thickness wounds created on the dorsal aspects of athymic mice. The monolayer CM promoted wound healing as compared with fresh medium or no treatment. Importantly, wound closure was faster, and dermal and epidermal regeneration was improved in the spheroid CM-treated mice compared with that in the monolayer CM-treated mice. The improved wound healing by spheroid CM may be attributed, at least in part, to enhanced neovascularization in the wound beds. The spheroid-based CM approach showed potential as a therapy for skin wound repair. Copyright © 2015 Elsevier Inc. All rights reserved.

  4. Conditioned Medium from Placental Mesenchymal Stem Cells Reduces Oxidative Stress during the Cryopreservation of Ex Vivo Expanded Umbilical Cord Blood Cells

    PubMed Central

    Kadekar, Darshana; Rangole, Sonal; Kale, Vaijayanti; Limaye, Lalita

    2016-01-01

    Background The limited cell dose in umbilical cord blood (UCB) necessitates ex vivo expansion of UCB. Further, the effective cryopreservation of these expanded cells is important in widening their use in the clinics. During cryopreservation, cells experience oxidative stress due to the generation of reactive oxygen species (ROS). Conditioned medium from mesenchymal stem cells (MSCs-CM) has been shown to alleviate the oxidative stress during wound healing, Alzheimer’s disease and ischemic disease. This premise prompted us to investigate the influence of MSCs-CM during cryopreservation of expanded UCB cells. Methodology/Principle findings CM-was collected from cord/placental MSCs(C-MSCs-CM, P-MSC-CM). UCB CD34+cells were expanded as suspension cultures in serum free medium containing cytokines for 10 days. Cells were frozen with/without C-MSCs-CM and or P-MSCs-CM in the conventional freezing medium containing 20%FCS +10%DMSO using a programmable freezer and stored in liquid nitrogen. Upon revival, cells frozen with MSCs-CM were found to be superior to cells frozen in conventional medium in terms of viability, CD34+content and clonogenecity. Priming of revived cells for 48 hrs with MSCs-CM further improved their transplantation ability, as compared to those cultured without MSCs-CM. P-MSCs-CM radically reduced the oxidative stress in cryopreserved cells, resulting in better post thaw functionality of CD34+ cells than with C-MSCs-CM. The observed cryoprotective effect of MSCs-CM was primarily due to anti-oxidative and anti-apoptotic properties of the MSCs-CM and not because of the exosomes secreted by them. Conclusions/Significance Our data suggest that MSCs-CM can serve as a valuable additive to the freezing or the priming medium for expanded UCB cells, which would increase their clinical applicability. PMID:27780236

  5. 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.

  6. Conditioned medium: a new alternative for cryopreservation of equine umbilical cord mesenchymal stem cells.

    PubMed

    Maia, Leandro; Dias, Marianne Camargos; de Moraes, Carolina Nogueira; de Paula Freitas-Dell'Aqua, Camila; da Mota, Ligia S L Silveira; Santiloni, Valquíria; da Cruz Landim-Alvarenga, Fernanda

    2017-03-01

    Cryopreservation is a feasible alternative to maintaining several cell lines, particularly for immediate therapeutic use, transportation of samples, and implementation of new in vitro studies. This work parts from the hypothesis that the medium of cryopreservation composed by 90% of conditioned medium (CM) supports cryopreservation of equine umbilical cord intervascular matrix mesenchymal stem cells (UCIM-MSCs), allowing the maintenance of the biological properties for the establishment of cell banks intended for therapeutic use and in vitro studies. Thus, we evaluated the viability, apoptosis/necrosis rates, immunophenotypic profile (IP), chromosomal stability, clonicity, and differentiation potential of UCIM-MSCs cryopreserved with four different mediums (with FBS: M1, M3, M4 and without FBS: M2). After 3 months of cryopreservation, samples were thawed and analyzed. The potential of differentiation in the mesodermal lineages, clonicity, and the chromosomal stability were maintained after cryopreservation of UCIM-MSCs with medium containing FBS. Changes (P < 0.05) at IP for some markers were observed at cells cryopreserved with medium M1-M3. Only the UCIM-MSCs cryopreserved with the CM (M4) had similar viability post-thaw (P = 0.23) when compared with fresh cells. We proved the hypothesis that the medium of cryopreservation containing CM supports the cryopreservation of UCIM-MSCs, at the experimental conditions, being the medium that better maintains the biological characteristics observed at fresh cells. Thus, future studies of UCIM-MSCs secretome should be conducted to better understand the beneficial and protective effects of the CM during the freezing process. © 2017 International Federation for Cell Biology.

  7. Adult hippocampus derived soluble factors induce a neuronal-like phenotype in mesenchymal stem cells.

    PubMed

    Rivera, Francisco J; Sierralta, Walter D; Minguell, Jose J; Aigner, Ludwig

    2006-10-02

    Bone marrow-derived mesenchymal stem cells (MSCs) are not restricted in their differentiation fate to cells of the mesenchymal lineage. They acquire a neural phenotype in vitro and in vivo after transplantation in the central nervous system. Here we investigated whether soluble factors derived from different brain regions are sufficient to induce a neuronal phenotype in MSCs. We incubated bone marrow-derived MSCs in conditioned medium (CM) derived from adult hippocampus (HCM), cortex (CoCM) or cerebellum (CeCM) and analyzed the cellular morphology and the expression of neuronal and glial markers. In contrast to muscle derived conditioned medium, which served as control, conditioned medium derived from the different brain regions induced a neuronal morphology and the expression of the neuronal markers GAP-43 and neurofilaments in MSCs. Hippocampus derived conditioned medium had the strongest activity. It was independent of NGF or BDNF; and it was restricted to the neuronal differentiation fate, since no induction of the astroglial marker GFAP was observed. The work indicates that soluble factors present in the brain are sufficient to induce a neuronal phenotype in MSCs.

  8. Delayed Administration of Bone Marrow Mesenchymal Stem Cell Conditioned Medium Significantly Improves Outcome After Retinal Ischemia in Rats

    PubMed Central

    Dreixler, John C.; Poston, Jacqueline N.; Balyasnikova, Irina; Shaikh, Afzhal R.; Tupper, Kelsey Y.; Conway, Sineadh; Boddapati, Venkat; Marcet, Marcus M.; Lesniak, Maciej S.; Roth, Steven

    2014-01-01

    Purpose. Delayed treatment after ischemia is often unsatisfactory. We hypothesized that injection of bone marrow stem cell (BMSC) conditioned medium after ischemia could rescue ischemic retina, and in this study we characterized the functional and histological outcomes and mechanisms of this neuroprotection. Methods. Retinal ischemia was produced in adult Wistar rats by increasing intraocular pressure for 55 minutes. Conditioned medium (CM) from rat BMSCs or unconditioned medium (uCM) was injected into the vitreous 24 hours after the end of ischemia. Recovery was assessed 7 days after ischemia using electroretinography, at which time we euthanized the animals and then prepared 4-μm-thick paraffin-embedded retinal sections. TUNEL and Western blot were used to identify apoptotic cells and apoptosis-related gene expression 24 hours after injections; that is, 48 hours after ischemia. Protein content in CM versus uCM was studied using tandem mass spectrometry, and bioinformatics methods were used to model protein interactions. Results. Intravitreal injection of CM 24 hours after ischemia significantly improved retinal function and attenuated cell loss in the retinal ganglion cell layer. CM attenuated postischemic apoptosis and apoptosis-related gene expression. By spectral counting, 19 proteins that met stringent identification criteria were increased in the CM compared to uCM; the majority were extracellular matrix proteins that mapped into an interactional network together with other proteins involved in cell growth and adhesion. Conclusions. By restoring retinal function, attenuating apoptosis, and preventing retinal cell loss after ischemia, CM is a robust means of delayed postischemic intervention. We identified some potential candidate proteins for this effect. PMID:24699381

  9. Adipose stem cells can secrete angiogenic factors that inhibit hyaline cartilage regeneration.

    PubMed

    Lee, Christopher Sd; Burnsed, Olivia A; Raghuram, Vineeth; Kalisvaart, Jonathan; Boyan, Barbara D; Schwartz, Zvi

    2012-08-24

    Adipose stem cells (ASCs) secrete many trophic factors that can stimulate tissue repair, including angiogenic factors, but little is known about how ASCs and their secreted factors influence cartilage regeneration. Therefore, the aim of this study was to determine the effects ASC-secreted factors have in repairing chondral defects. ASCs isolated from male Sprague Dawley rats were cultured in monolayer or alginate microbeads supplemented with growth (GM) or chondrogenic medium (CM). Subsequent co-culture, conditioned media, and in vivo cartilage defect studies were performed. ASC monolayers and microbeads cultured in CM had decreased FGF-2 gene expression and VEGF-A secretion compared to ASCs cultured in GM. Chondrocytes co-cultured with GM-cultured ASCs for 7 days had decreased mRNAs for col2, comp, and runx2. Chondrocytes treated for 12 or 24 hours with conditioned medium from GM-cultured ASCs had reduced sox9, acan, and col2 mRNAs; reduced proliferation and proteoglycan synthesis; and increased apoptosis. ASC-conditioned medium also increased endothelial cell tube lengthening whereas conditioned medium from CM-cultured ASCs had no effect. Treating ASCs with CM reduced or abolished these deleterious effects while adding a neutralizing antibody for VEGF-A eliminated ASC-conditioned medium induced chondrocyte apoptosis and restored proteoglycan synthesis. FGF-2 also mitigated the deleterious effects VEGF-A had on chondrocyte apoptosis and phenotype. When GM-grown ASC pellets were implanted in 1 mm non-critical hyaline cartilage defects in vivo, cartilage regeneration was inhibited as evaluated by radiographic and equilibrium partitioning of an ionic contrast agent via microCT imaging. Histology revealed that defects with GM-cultured ASCs had no tissue ingrowth from the edges of the defect whereas empty defects and defects with CM-grown ASCs had similar amounts of neocartilage formation. ASCs must be treated to reduce the secretion of VEGF-A and other factors that inhibit cartilage regeneration, which can significantly influence how ASCs are used for repairing hyaline cartilage.

  10. Conditioned medium from the three-dimensional culture of human umbilical cord perivascular cells accelerate the migration and proliferation of human keratinocyte and fibroblast.

    PubMed

    Kim, Min Ho; Wu, Wen Hao; Choi, Jee Hyun; Kim, Ji Hyun; Hong, Seok-Ho; Jun, Jin Hyun; Ko, Yong; Lee, Jong Hun

    Previous studies have reported that the conditioned medium (CM) of bone marrow-mesenchymal stem cells (BM-MSCs) stimulate the migration and proliferation of cell types involved in the wound healing process. However, these studies only show MSC-CM effects that were obtained using a two-dimensional (2D) culture. Recently, a three-dimensional (3D) culture has been considered to be a more physiologically appropriate system than the 2D culture. In addition, it has been shown that the procurement of BM-MSC is invasive, and other sources of MSC are thus being explored. Recently, perivascular cells (PVCs) have been considered as an alternative source of cells for dermal wound healing. Therefore, in this study, a PVC-conditioned medium (CM) was collected from a 3D culture (PVC-CM-3D) using highly porous polystyrene-based membranes and compared with PVC-CM from a 2D culture (PVC-CM-2D) to investigate the effects on the migration and proliferation of human keratinocytes and fibroblasts. Moreover, the PVC-CM components from the 2D and 3D cultures were identified using 2D gel electrophoresis. The migrations of the keratinocytes cells and fibroblasts were significantly higher with PVC-CM-3D than with the 2D culture; similarly, the proliferation of keratinocytes was also highly stimulated by PVC-CM-3D. Proteomic analyses of the PVC-CM revealed that type I collagen was highly expressed in the 3D-culture system. Microtubule-actin cross-linked factor 1 (KIAA0465), nebulin-related anchoring protein, and thioredoxin were specifically expressed only in PVC-CM-3D. In addition, more EVs could be isolated from the PVC-CM-3D, and EVs were found to stimulate keratinocyte migration. Taken together, 3D-culture using a polystyrene scaffold is demonstrated to be a better system for providing better physiological conditions; therefore, PVC-CM-3D could be a promising option for skin-wound healing.

  11. Chondrogenically primed tonsil-derived mesenchymal stem cells encapsulated in riboflavin-induced photocrosslinking collagen-hyaluronic acid hydrogel for meniscus tissue repairs.

    PubMed

    Koh, Rachel H; Jin, Yinji; Kang, Byung-Jae; Hwang, Nathaniel S

    2017-04-15

    Current meniscus tissue repairing strategies involve partial or total meniscectomy, followed by allograft transplantation or synthetic material implantation. However, allografts and synthetic implants have major drawbacks such as the limited supply of grafts and lack of integration into host tissue, respectively. In this study, we investigated the effects of conditioned medium (CM) from meniscal fibrochondrocytes and TGF-β3 on tonsil-derived mesenchymal stem cells (T-MSCs) for meniscus tissue engineering. CM-expanded T-MSCs were encapsulated in riboflavin-induced photocrosslinked collagen-hyaluronic acid (COL-RF-HA) hydrogels and cultured in chondrogenic medium containing TGF-β3. In vitro results indicate that CM-expanded cells followed by TGF-β3 exposure stimulated the expression of fibrocartilage-related genes (COL2, SOX9, ACAN, COL1) and production of extracellular matrix components. Histological assessment of in vitro and subcutaneously implanted in vivo constructs demonstrated that CM-expanded cells followed by TGF-β3 exposure resulted in highest cell proliferation, GAG accumulation, and collagen deposition. Furthermore, when implanted into meniscus defect model, CM treatment amplified the potential of TGF-β3 and induced complete regeneration. Conditioned medium derived from chondrocytes have been reported to effectively prime mesenchymal stem cells toward chondrogenic lineage. Type I collagen is the main component of meniscus extracellular matrix and hyaluronic acid is known to promote meniscus regeneration. In this manuscript, we investigated the effects of conditioned medium (CM) and transforming growth factor-β3 (TGF-β3) on tonsil-derived mesenchymal stem cells (T-MSCs) encapsulated in riboflavin-induced photocrosslinked collagen-hyaluronic acid (COL-RF-HA) hydrogel. We employed a novel source of conditioned medium, derived from meniscal fibrochondrocytes. Our in vitro and in vivo results collectively illustrate that CM-expanded cells followed by TGF-β3 exposure have the best potential for meniscus regeneration. This manuscript highlights a novel stem cell commitment strategy combined with biomaterials designs for meniscus regeneration. Copyright © 2017 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.

  12. Mesenchymal Stem Cell-Conditioned Medium Reduces Disease Severity and Immune Responses in Inflammatory Arthritis.

    PubMed

    Kay, Alasdair G; Long, Grace; Tyler, George; Stefan, Andrei; Broadfoot, Stephen J; Piccinini, Anna M; Middleton, Jim; Kehoe, Oksana

    2017-12-21

    We evaluated the therapeutic potential of mesenchymal stem cell-conditioned medium (CM-MSC) as an alternative to cell therapy in an antigen-induced model of arthritis (AIA). Disease severity and cartilage loss were evaluated by histopathological analysis of arthritic knee joints and immunostaining of aggrecan neoepitopes. Cell proliferation was assessed for activated and naïve CD4+ T cells from healthy mice following culture with CM-MSC or co-culture with MSCs. T cell polarization was analysed in CD4+ T cells isolated from spleens and lymph nodes of arthritic mice treated with CM-MSC or MSCs. CM-MSC treatment significantly reduced knee-joint swelling, histopathological signs of AIA, cartilage loss and suppressed TNFα induction. Proliferation of CD4+ cells from spleens of healthy mice was not affected by CM-MSC but reduced when cells were co-cultured with MSCs. In the presence of CM-MSC or MSCs, increases in IL-10 concentration were observed in culture medium. Finally, CD4+ T cells from arthritic mice treated with CM-MSC showed increases in FOXP3 and IL-4 expression and positively affected the Treg:Th17 balance in the tissue. CM-MSC treatment reduces cartilage damage and suppresses immune responses by reducing aggrecan cleavage, enhancing Treg function and adjusting the Treg:Th17 ratio. CM-MSC may provide an effective cell-free therapy for inflammatory arthritis.

  13. Three-dimensional spheroid cell culture of umbilical cord tissue-derived mesenchymal stromal cells leads to enhanced paracrine induction of wound healing.

    PubMed

    Santos, Jorge M; Camões, Sérgio P; Filipe, Elysse; Cipriano, Madalena; Barcia, Rita N; Filipe, Mariana; Teixeira, Mariana; Simões, Sandra; Gaspar, Manuela; Mosqueira, Diogo; Nascimento, Diana S; Pinto-do-Ó, Perpétua; Cruz, Pedro; Cruz, Helder; Castro, Matilde; Miranda, Joana P

    2015-05-09

    The secretion of trophic factors by mesenchymal stromal cells has gained increased interest given the benefits it may bring to the treatment of a variety of traumatic injuries such as skin wounds. Herein, we report on a three-dimensional culture-based method to improve the paracrine activity of a specific population of umbilical cord tissue-derived mesenchymal stromal cells (UCX®) towards the application of conditioned medium for the treatment of cutaneous wounds. A UCX® three-dimensional culture model was developed and characterized with respect to spheroid formation, cell phenotype and cell viability. The secretion by UCX® spheroids of extracellular matrix proteins and trophic factors involved in the wound-healing process was analysed. The skin regenerative potential of UCX® three-dimensional culture-derived conditioned medium (CM3D) was also assessed in vitro and in vivo against UCX® two-dimensional culture-derived conditioned medium (CM2D) using scratch and tubulogenesis assays and a rat wound splinting model, respectively. UCX® spheroids kept in our three-dimensional system remained viable and multipotent and secreted considerable amounts of vascular endothelial growth factor A, which was undetected in two-dimensional cultures, and higher amounts of matrix metalloproteinase-2, matrix metalloproteinase-9, hepatocyte growth factor, transforming growth factor β1, granulocyte-colony stimulating factor, fibroblast growth factor 2 and interleukin-6, when compared to CM2D. Furthermore, CM3D significantly enhanced elastin production and migration of keratinocytes and fibroblasts in vitro. In turn, tubulogenesis assays revealed increased capillary maturation in the presence of CM3D, as seen by a significant increase in capillary thickness and length when compared to CM2D, and increased branching points and capillary number when compared to basal medium. Finally, CM3D-treated wounds presented signs of faster and better resolution when compared to untreated and CM2D-treated wounds in vivo. Although CM2D proved to be beneficial, CM3D-treated wounds revealed a completely regenerated tissue by day 14 after excisions, with a more mature vascular system already showing glands and hair follicles. This work unravels an important alternative to the use of cells in the final formulation of advanced therapy medicinal products by providing a proof of concept that a reproducible system for the production of UCX®-conditioned medium can be used to prime a secretome for eventual clinical applications.

  14. Effect of photobiomodulation on viability and proliferation of stem cells from exfoliated deciduous teeth under different nutritional conditions

    NASA Astrophysics Data System (ADS)

    Morato de Souza, Letícia; Guilherme Roque Rinco, Ugo; Aparecida Tavares Aguiar, Daniela; Aparecido de Almeida Junior, Luciano; Cosme-Silva, Leopoldo; Marchini Oliveira, Thais; Teixeira Marques, Nádia Carolina; Thiemy Sakai, Vivien

    2018-02-01

    This study aimed to evaluate the effect of different doses of low-level laser irradiation on the viability and proliferation of stem cells from exfoliated deciduous teeth (SHED) cultured under nutritional deficit (cellular stress) or regular nutritional conditions. SHED underwent irradiation by a red laser between 1.2 and 6.2 J cm-2. Prior to the irradiation, all groups received culture medium (MEMα, Eagle’s minimum essential medium alpha modification) supplemented with 1% of fetal bovine serum (FBS) for 1 h. After the irradiation, cells received MEMα supplemented with 10% of FBS (regular nutrition) or 1% of FBS (nutritional deficit). Cell viability and proliferation were respectively determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and crystal violet assays 6 and 24 h after irradiation (P  <  0.05). At 24 h, SHED under nutritional deficit showed lower viability and proliferation after 1.2 J cm-2 irradiation. All of the irradiated groups revealed significantly higher viability and proliferation in SHED maintained under nutritional deficit than in regular nutritional conditions, except in the 3.7 and 6.2 J cm-2 groups by MTT assay. In the crystal violet assay, SHED irradiated with 1.2 J cm-2 showed no difference between the different nutritional conditions. Decrease of FBS concentration in the culture medium seems to enhance the sensitivity of SHED to the effects of photobiomodulation therapy. Nutritional stress conditions improved cell viability and proliferation of SHED after laser irradiation, except for 1.2 J cm-2.

  15. Effects of conditioned medium from LL-37 treated adipose stem cells on human fibroblast migration.

    PubMed

    Yang, Eun-Jung; Bang, Sa-Ik

    2017-07-01

    Adipose stem cell-conditioned medium may promote human dermal fibroblast (HDF) proliferation and migration by activating paracrine peptides during the re-epithelization phase of wound healing. Human antimicrobial peptide LL-37 is upregulated in the skin epithelium as part of the normal response to injury. The effects of conditioned medium (CM) from LL-37 treated adipose stem cells (ASCs) on cutaneous wound healing, including the mediation of fibroblast migration, remain to be elucidated, therefore the aim of the present study was to determine how ASCs would react to an LL-37-rich microenvironment and if CM from LL-37 treated ASCs may influence the migration of HDFs. The present study conducted migration assays with HDFs treated with CM from LL-37 treated ASCs. Expression of CXC chemokine receptor 4 (CXCR4), which controls the recruitment of HDFs, was analyzed at the mRNA and protein levels. To further characterize the stimulatory effects of LL-37 on ASCs, the expression of stromal cell-derived factor-1α (SDF-1α), a CXC chemokine, was investigated. CM from LL-37-treated ASCs induced migration of HDFs in a time- and dose-dependent manner, with a maximum difference in migration observed 24 h following stimulation with LL-37 at a concentration of 10 µg/ml. The HDF migration and the expression of CXCR4 in fibroblasts was markedly increased upon treatment with CM from LL-37-treated ASCs compared with CM from untreated ASCs. SDF-1α expression was markedly increased in CM from LL-37 treated ASCs. It was additionally observed that SDF-1α blockade significantly reduced HDF migration. These findings suggest the feasibility of CM from LL-37-treated ASCs as a potential therapeutic for human dermal fibroblast migration.

  16. Progranulin, a Major Secreted Protein of Mouse Adipose-Derived Stem Cells, Inhibits Light-Induced Retinal Degeneration

    PubMed Central

    Tsuruma, Kazuhiro; Yamauchi, Mika; Sugitani, Sou; Otsuka, Tomohiro; Ohno, Yuta; Nagahara, Yuki; Ikegame, Yuka; Shimazawa, Masamitsu; Yoshimura, Shinichi; Iwama, Toru

    2014-01-01

    Adipose tissue stromal vascular fraction contains mesenchymal stem cells, which show protective effects when administered to damaged tissues, mainly through secreted trophic factors. We examined the protective effects of adipose-derived stem cells (ASCs) and ASC-conditioned medium (ASC-CM) against retinal damage and identified the neuroprotective factors in ASC-CM. ASCs and mature adipocytes were isolated from mouse subcutaneous tissue. ASCs were injected intravitreally in a mouse model of light-induced retinal damage, and ASC injection recovered retinal function as measured by electroretinogram and inhibited outer nuclear layer, thinning, without engraftment of ASCs. ASC-CM and mature adipocyte-conditioned medium were collected after 72 hours of culture. In vitro, H2O2- and light-induced cell death was reduced in a photoreceptor cell line with ASC-CM but not with mature adipocyte-conditioned medium. In vivo, light-induced photoreceptor damage was evaluated by measurement of outer nuclear layer thickness at 5 days after light exposure and by electroretinogram recording. ASC-CM significantly inhibited photoreceptor degeneration and retinal dysfunction after light exposure. Progranulin was identified as a major secreted protein of ASCs that showed protective effects against retinal damage in vitro and in vivo. Furthermore, progranulin phosphorylated extracellular signal-regulated kinase, cAMP response element binding protein, and hepatocyte growth factor receptor, and protein kinase C signaling pathways were involved in the protective effects of progranulin. These findings suggest that ASC-CM and progranulin have neuroprotective effects in the light-induced retinal-damage model. Progranulin may be a potential target for the treatment of the degenerative diseases of the retina. PMID:24233842

  17. Progranulin, a major secreted protein of mouse adipose-derived stem cells, inhibits light-induced retinal degeneration.

    PubMed

    Tsuruma, Kazuhiro; Yamauchi, Mika; Sugitani, Sou; Otsuka, Tomohiro; Ohno, Yuta; Nagahara, Yuki; Ikegame, Yuka; Shimazawa, Masamitsu; Yoshimura, Shinichi; Iwama, Toru; Hara, Hideaki

    2014-01-01

    Adipose tissue stromal vascular fraction contains mesenchymal stem cells, which show protective effects when administered to damaged tissues, mainly through secreted trophic factors. We examined the protective effects of adipose-derived stem cells (ASCs) and ASC-conditioned medium (ASC-CM) against retinal damage and identified the neuroprotective factors in ASC-CM. ASCs and mature adipocytes were isolated from mouse subcutaneous tissue. ASCs were injected intravitreally in a mouse model of light-induced retinal damage, and ASC injection recovered retinal function as measured by electroretinogram and inhibited outer nuclear layer, thinning, without engraftment of ASCs. ASC-CM and mature adipocyte-conditioned medium were collected after 72 hours of culture. In vitro, H2O2- and light-induced cell death was reduced in a photoreceptor cell line with ASC-CM but not with mature adipocyte-conditioned medium. In vivo, light-induced photoreceptor damage was evaluated by measurement of outer nuclear layer thickness at 5 days after light exposure and by electroretinogram recording. ASC-CM significantly inhibited photoreceptor degeneration and retinal dysfunction after light exposure. Progranulin was identified as a major secreted protein of ASCs that showed protective effects against retinal damage in vitro and in vivo. Furthermore, progranulin phosphorylated extracellular signal-regulated kinase, cAMP response element binding protein, and hepatocyte growth factor receptor, and protein kinase C signaling pathways were involved in the protective effects of progranulin. These findings suggest that ASC-CM and progranulin have neuroprotective effects in the light-induced retinal-damage model. Progranulin may be a potential target for the treatment of the degenerative diseases of the retina.

  18. Conditioned Medium from Early-Outgrowth Bone Marrow Cells Is Retinal Protective in Experimental Model of Diabetes

    PubMed Central

    Duarte, Diego A.; Papadimitriou, Alexandros; Gilbert, Richard E.; Thai, Kerri; Zhang, Yanling; Rosales, Mariana A. B.; Lopes de Faria, José B.; Lopes de Faria, Jacqueline M.

    2016-01-01

    Bone marrow-derived cells were demonstrated to improve organ function, but the lack of cell retention within injured organs suggests that the protective effects are due to factors released by the cells. Herein, we tested cell therapy using early outgrowth cells (EOCs) or their conditioned media (CM) to protect the retina of diabetic animal models (type 1 and type 2) and assessed the mechanisms by in vitro study. Control and diabetic (db/db) mice (8 weeks of age) were randomized to receive a unique intravenous injection of 5×105EOCs or 0.25 ml thrice weekly tail-vein injections of 10x concentrated CM and Wystar Kyoto rats rendered diabetic were randomized to receive 0.50 ml thrice weekly tail-vein injections of 10x concentrated CM. Four weeks later, the animals were euthanized and the eyes were enucleated. Rat retinal Müller cells (rMCs) were exposed for 24 h to high glucose (HG), combined or not with EOC-conditioned medium (EOC-CM) from db/m EOC cultures. Diabetic animals showed increase in diabetic retinopathy (DR) and oxidative damage markers; the treatment with EOCs or CM infusions significantly reduced this damage and re-established the retinal function. In rMCs exposed to diabetic milieu conditions (HG), the presence of EOC-CM reduced reactive oxygen species production by modulating the NADPH-oxidase 4 system, thus upregulating SIRT1 activity and deacetylating Lys-310-p65-NFκB, decreasing GFAP and VEGF expressions. The antioxidant capacity of EOC-CM led to the prevention of carbonylation and nitrosylation posttranslational modifications on the SIRT1 molecule, preserving its activity. The pivotal role of SIRT1 on the mode of action of EOCs or their CM was also demonstrated on diabetic retina. These findings suggest that EOCs are effective as a form of systemic delivery for preventing the early molecular markers of DR and its conditioned medium is equally protective revealing a novel possibility for cell-free therapy for the treatment of DR. PMID:26836609

  19. Orion Landing Simulation Eight Soil Model Comparison

    NASA Technical Reports Server (NTRS)

    Mark, Stephen D.

    2009-01-01

    LS-DYNA finite element simulations of a rigid Orion Crew Module (CM) were used to investigate the CM impact behavior on eight different soil models. Ten different landing conditions, characterized by the combination of CM vertical and horizontal velocity, hang angle, and roll angle were simulated on the eight different soils. The CM center of gravity accelerations, pitch angle, kinetic energy, and soil contact forces were the outputs of interest. The simulation results are presented, with comparisons of the CM behavior on the different soils. The soils analyzed in this study can be roughly categorized as soft, medium, or hard, according to the CM accelerations that occur when landing on them. The soft group is comprised of the Carson Sink Wet soil and the Kennedy Space Center (KSC) Low Density Dry Sand. The medium group includes Carson Sink Dry, the KSC High Density In-Situ Moisture Sand and High Density Flooded Sand, and Cuddeback B. The hard soils are Cuddeback A and the Gantry Unwashed Sand. The softer soils were found to produce lower peak accelerations, have more stable pitch behavior, and to be less sensitive to the landing conditions. This investigation found that the Cuddeback A soil produced the highest peak accelerations and worst stability conditions, and that the best landing performance was achieved on the KSC Low Density Dry Sand.

  20. Novel application of stem cell-derived factors for periodontal regeneration

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

    Inukai, Takeharu, E-mail: t-inukai@med.nagoya-u.ac.jp; Katagiri, Wataru, E-mail: w-kat@med.nagoya-u.ac.jp; Yoshimi, Ryoko, E-mail: lianzi@med.nagoya-u.ac.jp

    Highlights: Black-Right-Pointing-Pointer Mesenchymal stem cells (MSCs) secrete a variety of cytokines. Black-Right-Pointing-Pointer Cytokines were detected in conditioned medium from cultured MSCs (MSC-CM). Black-Right-Pointing-Pointer MSC-CM enhanced activation of dog MSCs and periodontal ligament cells. Black-Right-Pointing-Pointer MSC-CM significantly promoted alveolar bone and cementum regeneration. Black-Right-Pointing-Pointer Multiple cytokines contained in MSC-CM promote periodontal regeneration. -- Abstract: The effect of conditioned medium from cultured mesenchymal stem cells (MSC-CM) on periodontal regeneration was evaluated. In vitro, MSC-CM stimulated migration and proliferation of dog MSCs (dMSCs) and dog periodontal ligament cells (dPDLCs). Cytokines such as insulin-like growth factor, vascular endothelial growth factor, transforming growth factor-{beta}1, andmore » hepatocyte growth factor were detected in MSC-CM. In vivo, one-wall critical-size, intrabony periodontal defects were surgically created in the mandible of dogs. Dogs with these defects were divided into three groups that received MSC-CM, PBS, or no implants. Absorbable atelo-collagen sponges (TERUPLUG Registered-Sign ) were used as a scaffold material. Based on radiographic and histological observation 4 weeks after transplantation, the defect sites in the MSC-CM group displayed significantly greater alveolar bone and cementum regeneration than the other groups. These findings suggest that MSC-CM enhanced periodontal regeneration due to multiple cytokines contained in MSC-CM.« less

  1. Basic FGF Support of Human Embryonic Stem Cell Self-Renewal

    PubMed Central

    Levenstein, Mark E.; Ludwig, Tenneille E.; Xu, Ren-He; Llanas, Rachel A.; VanDenHeuvel-Kramer, Kaitlyn; Manning, Daisy; Thomson, James A.

    2015-01-01

    Human embryonic stem (ES) cells have most commonly been cultured in the presence of basic FGF (FGF2) either on fibroblast feeder layers or in fibroblast-conditioned medium. Recently, it has been reported that elevated concentrations of FGF2 permit the culture of human ES cells in the absence of fibroblasts or fibroblast-conditioned medium. Here we compare the ability of unconditioned medium (UM) supplemented with 4, 24, 40, 80, 100 and 250 ng/ml FGF2 to sustain low-density human ES cell cultures through multiple passages. In these stringent culture conditions, 4, 24, and 40 ng/ml FGF2 failed to sustain human ES cells through three passages, but 100 ng/ml sustained human ES cells with an effectiveness comparable to conditioned medium (CM). Two human ES cell lines (H1 and H9) were maintained for up to 164 population doublings (7 and 4 months) in UM supplemented with 100 ng/ml FGF2. After prolonged culture the cells formed teratomas when injected into SCID-beige mice, and expressed markers characteristic of undifferentiated human ES cells. We also demonstrate that FGF2 is degraded more rapidly in UM than in CM, partly explaining the need for higher concentrations of FGF2 in UM. These results further facilitate the large-scale, routine culture of human ES cells, and suggest that fibroblasts and fibroblast-conditioned medium sustain human ES cells in part by stabilizing FGF signaling above a critical threshold. PMID:16282444

  2. EDA-containing fibronectin increases proliferation of embryonic stem cells.

    PubMed

    Losino, Noelia; Waisman, Ariel; Solari, Claudia; Luzzani, Carlos; Espinosa, Darío Fernández; Sassone, Alina; Muro, Andrés F; Miriuka, Santiago; Sevlever, Gustavo; Barañao, Lino; Guberman, Alejandra

    2013-01-01

    Embryonic stem cells (ESC) need a set of specific factors to be propagated. They can also grow in conditioned medium (CM) derived from a bovine granulosa cell line BGC (BGC-CM), a medium that not only preserves their main features but also increases ESC´s proliferation rate. The mitogenic properties of this medium were previously reported, ascribing this effect to an alternative spliced generated fibronectin isoform that contains the extra domain A (FN EDA(+)). Here, we investigated if the FN EDA(+) isoform increased proliferation of mouse and human ES cells. We analyzed cell proliferation using conditioned media produced by different mouse embryonic fibroblast (MEF) lines genetically engineered to express FN constitutively including or excluding the EDA domain (FN EDA(-)), and in media supplemented with recombinant peptides containing or not the EDA. We found that the presence of EDA in the medium increased mouse and human ESC's proliferation rate. Here we showed for the first time that this FN isoform enhances ESC's proliferation. These findings suggest a possible conserved behavior for regulation of ES cells proliferation by this FN isoform and could contribute to improve their culturing conditions both for research and cell therapy.

  3. EDA-Containing Fibronectin Increases Proliferation of Embryonic Stem Cells

    PubMed Central

    Losino, Noelia; Waisman, Ariel; Solari, Claudia; Luzzani, Carlos; Espinosa, Darío Fernández; Sassone, Alina; Muro, Andrés F.; Miriuka, Santiago; Sevlever, Gustavo; Barañao, Lino; Guberman, Alejandra

    2013-01-01

    Embryonic stem cells (ESC) need a set of specific factors to be propagated. They can also grow in conditioned medium (CM) derived from a bovine granulosa cell line BGC (BGC-CM), a medium that not only preserves their main features but also increases ESC´s proliferation rate. The mitogenic properties of this medium were previously reported, ascribing this effect to an alternative spliced generated fibronectin isoform that contains the extra domain A (FN EDA+). Here, we investigated if the FN EDA+ isoform increased proliferation of mouse and human ES cells. We analyzed cell proliferation using conditioned media produced by different mouse embryonic fibroblast (MEF) lines genetically engineered to express FN constitutively including or excluding the EDA domain (FN EDA-), and in media supplemented with recombinant peptides containing or not the EDA. We found that the presence of EDA in the medium increased mouse and human ESC’s proliferation rate. Here we showed for the first time that this FN isoform enhances ESC’s proliferation. These findings suggest a possible conserved behavior for regulation of ES cells proliferation by this FN isoform and could contribute to improve their culturing conditions both for research and cell therapy. PMID:24244705

  4. Importance of dosimetry protocol for cell irradiation on a low X-rays facility and consequences for the biological response.

    PubMed

    Dos Santos, Morgane; Paget, Vincent; Ben Kacem, Mariam; Trompier, François; Benadjaoud, Mohamed Amine; François, Agnès; Guipaud, Olivier; Benderitter, Marc; Milliat, Fabien

    2018-06-01

    The main objective of radiobiology is to establish links between doses and radiation-induced biological effects. In this context, well-defined dosimetry protocols are crucial to the determination of experimental protocols. This work proposes a new dosimetry protocol for cell irradiation in a SARRP and shows the importance of the modification of some parameters defined in dosimetry protocol for physical dose and biological outcomes. Once all parameters of the configuration were defined, dosimetry measurements with ionization chambers and EBT3 films were performed to evaluate the dose rate and the attenuation due to the cell culture medium. To evaluate the influence of changes in cell culture volume and/or additional filtration, 6-well plates containing EBT3 films with water were used to determine the impact on the physical dose at 80 kV. Then, experiments with the same irradiation conditions were performed by replacing EBT3 films by HUVECs. The biological response was assessed using clonogenic assay. Using a 0.15 mm copper filter lead to a variation of +1% using medium thickness of 0.104 cm to -8% using a medium thickness of 0.936 cm on the physical dose compare to the reference condition (0.313 cm). For the 1 mm aluminum filter, a variation of +8 to -40% for the same medium thickness conditions has been observed. Cells irradiated in the same conditions showed significant differences in survival fraction, corroborating the effects of dosimetric changes on physical dose. This work shows the importance of dosimetry in radiobiology studies and the need of an accurate description of the dosimetry protocol used for irradiation.

  5. Conditioned media from differentiating craniofacial bone marrow stromal cells influence mineralization and proliferation in periodontal ligament stem cells.

    PubMed

    Jin, Zhenyu; Feng, Yuan; Liu, Hongwei

    2016-10-01

    Previous reports have mainly focused on the behavioral responses of human periodontal ligament stem cells (hPDLSCs) in interaction with tibia bone marrow stromal cells (BMSCs). However, there is little study on the biologic features of hPDLSCs under the induction of maxilla BMSCs (M-BMSCs) at different phases of osteogenic differentiation. We hypothesized that M-BMSCs undergoing osteogenic differentiation acted on the proliferation, differentiation, and bone-forming capacity of hPDLSCs. In this paper, primary hPDLSCs and human M-BMSCs (hM-BMSCs) were expanded in vitro. After screening of surface markers for characterization, hPDLSCs were cocultured with different phases of differentiating hM-BMSCs. Cell proliferation and alkaline phosphatase activity were examined, and mineralization-associated markers such as osteocalcin and runt-related transcription factor 2 of hPDLSCs in coculture with uninduced/osteoinduced hM-BMSCs were evaluated. hPDLSCs in hM-BMSCs-conditioned medium (hM-BMSCs-CM) group showed a reduction in proliferation compared with untreated hPDLSCs, while osteoinduced hM-BMSCs for 10 day-conditioned medium (hM-BMSCs-CM-10ds) and osteoinduced hM-BMSCs for 15 day-conditioned medium (hM-BMSCs-CM-15ds) enhance the proliferation of hPDLSCs. hM-BMSCs of separate differentiation stages temporarily inhibited osteogenesis of hPDLSCs in the early days. Upon extending time periods, uninduced/osteoinduced hM-BMSCs markedly enhanced osteogenesis of hPDLSCs to different degrees. The transplantation results showed hM-BMSCs-CM-15ds treatment promoted tissue regeneration to generate cementum/periodontal ligament-like structure characterized by hard-tissue formation. This research supported the notion that hM-BMSCs triggered osteogenesis of hPDLSCs suggesting important implications for periodontal engineering.

  6. TRLFS study on the complexation of Cm(III) and Eu(III) with SO3-Ph-BTBP.

    PubMed

    Wagner, Christoph; Müllich, Udo; Geist, Andreas; Panak, Petra J

    2015-10-21

    The complexation of Cm(iii) and Eu(iii) with a water soluble BTBP (sodium 3,3',3'',3'''-([2,2'-bipyridine]-6,6'-diylbis(1,2,4-triazine-3,5,6-triyl))tetrabenzenesulfonate, SO3-Ph-BTBP) is studied using time resolved laser fluorescence spectroscopy. For the complexation of Cm(iii) the influence of the medium (10(-3) M HClO4→ 0.5 M HNO3) is investigated in detail revealing important impacts of the applied medium (pH, ionic strength, anions) on the speciation and conditional stability constants. SO3-Ph-BTBP forms 1 : 2 complexes with Cm(iii) and Eu(iii). The conditional stability constants of [Cm(SO3-Ph-BTBP)2](5-) and [Eu(SO3-Ph-BTBP)2](5-) in 0.5 M HNO3 are determined to be log β02 = 7.3 ± 0.3 and log β02 = 5.4 ± 0.5, respectively. The difference of 1.9 orders of magnitude is in line with hydrophobic BT(B)P type ligands and shows that the selectivity is not affected by tuning the hydrophilicity using SO3-Ph-side chains.

  7. CD147 deficiency blocks IL-8 secretion and inhibits lung cancer-induced osteoclastogenesis

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

    Wang, Hongkai; Zhuo, Yunyun; Hu, Xu

    2015-03-06

    Bone is a frequent target of lung cancer metastasis, which is associated with significant morbidity and poor prognosis; however, the molecular basis of this process is still unknown. This study investigated the role of extracellular matrix metalloproteinase inducer (also known as cluster of differentiation (CD)147) in osteoclastogenesis resulting from bone metastasis, based on the enrichment of this glycoprotein on the surface of many malignant bone tumors. RNA interference was used to silence CD147 expression in A549 human lung cancer cells. Compared with conditioned medium (CM) from control cells (A549-CM), CM from CD147-deficient cells (A549-si-CM) suppressed receptor activator of nuclear factormore » κB ligand-stimulated osteoclastogenesis in RAW 264.7 cells and bone marrow-derived macrophages. The mRNA levels of osteoclast-specific genes such as tartrate-resistant acid phosphatase, calcitonin receptor, and cathepsin K were also reduced in the presence of A549-si-CM. CD147 knockdown in A549 cells decreased interleukin (IL)-8mRNA and protein expression. IL-8 is present in large amounts in A549-CM and mimicked its inductive effect on osteoclastogenesis; this was reversed by depletion of IL-8 from the medium. Taken together, these results indicate that CD147 promotes lung cancer-induced osteoclastogenesis by modulating IL-8 secretion, and suggest that CD147 is a potential therapeutic target for cancer-associated bone resorption in lung cancer patients. - Highlights: • Bone loss frequently results from lung cancer metastasis. • Cluster of differentiation (CD)147 was depleted in A549 lung adenocarcinoma cells. • RAW 264.7 cell osteoclastogenesis was blocked by medium from CD147-deficient cells. • Interleukin (IL)-8 level was reduced in the conditioned medium. • Osteoclastogenesis induced by lung tumor cells requires CD147-mediated IL-8 release.« less

  8. Conditioned medium from the stem cells of human dental pulp improves cognitive function in a mouse model of Alzheimer's disease.

    PubMed

    Mita, Tsuneyuki; Furukawa-Hibi, Yoko; Takeuchi, Hideyuki; Hattori, Hisashi; Yamada, Kiyofumi; Hibi, Hideharu; Ueda, Minoru; Yamamoto, Akihito

    2015-10-15

    Alzheimer's disease (AD) is a progressive, neurodegenerative disease characterized by a decline in cognitive abilities and the appearance of β-amyloid plaques in the brain. Although the pathogenic mechanisms associated with AD are not fully understood, activated microglia releasing various neurotoxic factors, including pro-inflammatory cytokines and oxidative stress mediators, appear to play major roles. Here, we investigated the therapeutic benefits of a serum-free conditioned medium (CM) derived from the stem cells of human exfoliated deciduous teeth (SHEDs) in a mouse model of AD. The intranasal administration of SHEDs in these mice resulted in substantially improved cognitive function. SHED-CM contained factors involved in multiple neuroregenerative mechanisms, such as neuroprotection, axonal elongation, neurotransmission, the suppression of inflammation, and microglial regulation. Notably, SHED-CM attenuated the pro-inflammatory responses induced by β-amyloid plaques, and generated an anti-inflammatory/tissue-regenerating environment, which was accompanied by the induction of anti-inflammatory M2-like microglia. Our data suggest that SHED-CM may provide significant therapeutic benefits for AD. Copyright © 2015 Elsevier B.V. All rights reserved.

  9. In vitro culture of bovine embryos in murine ES cell conditioned media negatively affects expression of pluripotency-related markers OCT4, SOX2 and SSEA1.

    PubMed

    Oliveira, C S; de Souza, M M; Saraiva, N Z; Tetzner, T A D; Lima, M R; Lopes, F L; Garcia, J M

    2012-06-01

    Despite extensive efforts, establishment of bovine embryonic stem (ES) cell lines has not been successful. We hypothesized that culture conditions for in vitro-produced (IVP) embryos, the most used source of inner cell mass (ICM) to obtain ES cells, might affect their undifferentiated state. Therefore, the aim of this work was to improve pluripotency of IVP blastocysts to produce suitable ICM for further culturing. We tested KSR and foetal calf serum (FCS) supplements in SOF medium and ES cell conditioned medium (CM) on IVC (groups: KSR, KSR CM, FCS and FCS CM). Cleavage and blastocyst rates were similar between all groups. Also, embryonic quality, assessed by apoptosis rates (TUNEL assay), total cell number and ICM percentage did not differ between experimental groups. However, expression of pluripotency-related markers was affected. We detected down-regulation of OCT3/4, SOX2 and SSEA1 in ICM of FCS CM blastocysts (p < 0.05). SOX2 gene expression revealed lower levels (p < 0.05) on KSR CM blastocysts and a remarkable variation in SOX2 mRNA levels on FCS-supplemented blastocysts. In conclusion, pluripotency-related markers tend to decrease after supplementation with ES cell CM, suggesting different mechanisms regulating mouse and bovine pluripotency. KSR supplementation did not differ from FCS, but FCS replacement by KSR may produce blastocysts with stable SOX2 gene expression levels. © 2011 Blackwell Verlag GmbH.

  10. Hair-growth-promoting effect of conditioned medium of high integrin α6 and low CD 71 (α6bri/CD71dim) positive keratinocyte cells.

    PubMed

    Won, Chong Hyun; Jeong, Yun-Mi; Kang, Sangjin; Koo, Tae-Sung; Park, So-Hyun; Park, Ki-Young; Sung, Young-Kwan; Sung, Jong-Hyuk

    2015-02-19

    Keratinocyte stem/progenitor cells (KSCs) reside in the bulge region of the hair follicles and may be involved in hair growth. Hair follicle dermal papilla cells (HFDPCs) and outer root sheath (ORS) cells were treated with conditioned medium (CM) of KSCs. Moreover, the effects of KSC-CM on hair growth were examined ex vivo and in vivo. A human growth factor chip array and RT-PCR were employed to identify enriched proteins in KSC-CM as compared with CM from keratinocytes. KSC-CM significantly increased the proliferation of HFDPCs and ORS cells, and increased the S-phase of the cell cycle in HFDPCs. KSC-CM led to the phosphorylation of ATK and ERK1/2 in both cell types. After subcutaneous injection of KSC-CM in C3H/HeN mice, a significant increase in hair growth and increased proliferation of hair matrix keratinocytes ex vivo was observed. We identified six proteins enriched in KSC-CM (amphiregulin, insulin-like growth factor binding protein-2, insulin-like growth factor binding protein-5, granulocyte macrophage-colony stimulating factor, Platelet-derived growth factor-AA, and vascular endothelial growth factor). A growth-factor cocktail that contains these six recombinant growth factors significantly increased the proliferation of HFDPCs and ORS cells and enhanced the hair growth of mouse models. These results collectively indicate that KSC-CM has the potential to increase hair growth via the proliferative capacity of HFDPCs and ORS cells.

  11. Oxaloacetate and adipose stromal cells-conditional medium synergistically protected potassium/serum deprivation-induced neuronal apoptosis.

    PubMed

    Liu, Qingpeng; Zhao, Gang; Zhou, Changwei; Farlow, Martin R; Du, Yansheng; Xu, Guangxu; Gu, Huiying

    2017-01-01

    Adipose stromal cells conditioned media (ASC-CM) protect neurons in a variety of neuronal death models including potassium/serum deprivation-induced neuronal apoptosis. In this study, we found that ASC-CM contained glutamate oxaloacetate transaminase and its substrate, oxaloacetate (OAA) directly protected cerebellar granule neurons (CGN) from apoptosis induced by serum and potassium deprivation. Additionally, OAA inhibited serum and potassium deprivation-induced caspase 3 activation. ASC-CM and OAA in combination had a synergistic neuroprotective effect. Clearly, different from ASC-CM-induced neuroprotection, OAA-induced neuroprotection was Akt- independent but JNK-dependent. These data establish a mechanistic basis supporting that the application of ASC-CM for neuroprotective treatments could be significantly enhanced by addition of OAA. Copyright © 2016 Elsevier Inc. All rights reserved.

  12. Simulations on false gain in recombination-pumped soft-X-ray lasers

    NASA Astrophysics Data System (ADS)

    Ozaki, T.; Kuroda, H.

    1997-10-01

    Numerical investigations are performed on false gain due to axial plasma expansion, which is expected to be important in initial proof-of-principle studies of recombination-pumped soft-X-ray lasers with extended capabilities. Modelling calculations of experiments with slab boron nitride targets reveal large false gain coefficients approaching 20 cm-1 in the case of plasmas with short active medium lengths. The false gain in the case of fiber targets is found to be of equal magnitude to that for slabs in the case of plasmas with less than 0.1 cm active medium lengths. Calculations for slab targets predict that adopting a tolerance of ǃ cm-1 for gain will severely restrict the time and the active medium length of the plasma that can be used for error-free observations, while those for fiber targets are found to be considerably relaxed. The effects of false gain in the 54.2 + Na Balmer ! laser is also investigated, again revealing the importance of this phenomena under optimum gain conditions.

  13. [Periodontal ligament regeneration using apical tooth germ cell-conditioned medium induced periodontal ligament cells sheet between dental tube and ceramic biologic bone].

    PubMed

    Ning, Huiying; Liu, Hongwei

    2011-08-01

    The purpose of this study was to establish an indirect co-culture system of rat apical tooth germ-conditioned medium (APTG-CM) and periodontal ligament cells (PDLCs). PDLCs were isolated and cultured through the method of enzyme-digestion. Vimentin and cytokeratin(CK) were used to demonstrate the cells' mesenchymal derivation. Co-culture system of APTG-CM and PDLCs for 28 days, osteocalcin (OCN), collagen type I (COL I) and bone sialoprotein (BSP) were detected in PDLCs by immunocytochemistry. Morphological changes were observed by inverted microscope. With building a transplant by dental tube, periodontal ligament cell sheet and ceramic biologic bone (CBB) in vitro, then, the combinations of dental tube and PDLCs incubated by APTG-CM were implanted subcutaneously into athymic mice for 8 weeks. This study demonstrated that cellular cementum-like tissue formed along the dentin surface and CBB, with fibrous tissue adjacent or inserted into CBB in vivo. PDLCs were grown better in the CBB than in dentin tubes. And the vertical fibers can't embed in the control. PDLCs, embedded within this APTG-CM, exhibite several phenotypic characteristics of cementoblast lineages. Thereby it contributes to the main processes of periodontal tissue regeneration with rat APTG-CM.

  14. Hair-Growth-Promoting Effect of Conditioned Medium of High Integrin α6 and Low CD 71 (α6bri/CD71dim) Positive Keratinocyte Cells

    PubMed Central

    Won, Chong Hyun; Jeong, Yun-Mi; Kang, Sangjin; Koo, Tae-Sung; Park, So-Hyun; Park, Ki-Young; Sung, Young-Kwan; Sung, Jong-Hyuk

    2015-01-01

    Keratinocyte stem/progenitor cells (KSCs) reside in the bulge region of the hair follicles and may be involved in hair growth. Hair follicle dermal papilla cells (HFDPCs) and outer root sheath (ORS) cells were treated with conditioned medium (CM) of KSCs. Moreover, the effects of KSC-CM on hair growth were examined ex vivo and in vivo. A human growth factor chip array and RT-PCR were employed to identify enriched proteins in KSC-CM as compared with CM from keratinocytes. KSC-CM significantly increased the proliferation of HFDPCs and ORS cells, and increased the S-phase of the cell cycle in HFDPCs. KSC-CM led to the phosphorylation of ATK and ERK1/2 in both cell types. After subcutaneous injection of KSC-CM in C3H/HeN mice, a significant increase in hair growth and increased proliferation of hair matrix keratinocytes ex vivo was observed. We identified six proteins enriched in KSC-CM (amphiregulin, insulin-like growth factor binding protein-2, insulin-like growth factor binding protein-5, granulocyte macrophage-colony stimulating factor, Platelet-derived growth factor-AA, and vascular endothelial growth factor). A growth-factor cocktail that contains these six recombinant growth factors significantly increased the proliferation of HFDPCs and ORS cells and enhanced the hair growth of mouse models. These results collectively indicate that KSC-CM has the potential to increase hair growth via the proliferative capacity of HFDPCs and ORS cells. PMID:25706512

  15. Short-term preconditioning enhances the therapeutic potential of adipose-derived stromal/stem cell-conditioned medium in cisplatin-induced acute kidney injury.

    PubMed

    Overath, Jürgen M; Gauer, Stefan; Obermüller, Nicholas; Schubert, Ralf; Schäfer, Richard; Geiger, Helmut; Baer, Patrick C

    2016-03-15

    The development of new strategies to preserve renal function after acute kidney injury (AKI) is necessary due to limited clinical intervention options. The organ-protective effects of mesenchymal stromal/stem cells (MSCs) and their conditioned medium (CM) have been investigated demonstrating that both separately promoted tubular recovery and ameliorated the outcome of AKI. Nevertheless, strategies to optimise the regenerative potential of both are highly needed. Here we investigated the effects of CM from adipose-derived MSCs (ASCs) preincubated in a hypoxic environment (Hyp). Protective factors were investigated by PCR analysis and a protein array in vitro. The expression of 64 of the 308 proteins assayed was found to be more than two-fold increased after Hyp. CM of Hyp-pretreated ASCs (pCM) was used to enhance regeneration in a mouse model of cisplatin-induced AKI (cisAKI). Renal function was assessed by measurements of markers for AKI and serum cytokine levels. The pCM significantly ameliorated serum creatinine and neutrophil gelatinase-associated lipocalin values, and also the levels of inflammatory cytokines IL-1β and IL-6 in the serum of mice with AKI. Our work clearly showed that a Hyp preconditioning significantly increases the release of protective factors in ASCs and enhances the therapeutic effects of CM in cisAKI in mice. Copyright © 2016 Elsevier Inc. All rights reserved.

  16. Conditioned medium of dental pulp cells stimulated by Chinese propolis show neuroprotection and neurite extension in vitro.

    PubMed

    Kudo, Daichi; Inden, Masatoshi; Sekine, Shin-Ichiro; Tamaoki, Naritaka; Iida, Kazuki; Naito, Eiji; Watanabe, Kazuhiro; Kamishina, Hiroaki; Shibata, Toshiyuki; Hozumi, Isao

    2015-03-04

    The purpose of this study was to clarify the effect of Chinese propolis on the expression level of neurotrophic factors in dental pulp cells (DPCs). We also investigated that the effects of the conditioned medium (CM) of DPCs stimulated by the propolis against oxidative and endoplasmic reticulum (ER) stresses in human neuroblastoma SH-SY5Y cells, and on neurite extensions in rat adrenal pheochromocytoma PC12 cells. To investigate the effect of the propolis on the levels of neurotrophic factors in DPCs, we performed a qRT-PCR experiment. As results, NGF, but not BDNF and NT-3, in DPCs was significantly elevated by the propolis in a concentration-dependent manner. H2O2-induced cell death was significantly inhibited by the treatment with the CM of DPCs. In addition, the treatment with the propolis-stimulated CM of DPCs had a more protective effect than that with the CM of DPCs. We also examine the effect of the propolis-stimulated CM of DPCs against a tunicamycin-induced ER stress. The treatment with the propolis-stimulated CM as well as the CM of DPCs significantly inhibited tunicamycin-induced cell death. Moreover, the treatment with the propolis-stimulated CM of DPCs significantly induced neurite outgrowth from PC12 cells than that with the CM of DPCs. These results suggest that the CM of DPCs as well as DPCs will be an efficient source of new treatments for neurodegenerative diseases and that the propolis promote the advantage of the CM of DPCs via producing neurotrophic factors. Copyright © 2015 Elsevier Ireland Ltd. All rights reserved.

  17. Anti-inflammatory effect of conditioned medium from human uterine cervical stem cells in uveitis.

    PubMed

    Bermudez, Maria A; Sendon-Lago, Juan; Seoane, Samuel; Eiro, Noemi; Gonzalez, Francisco; Saa, Jorge; Vizoso, Francisco; Perez-Fernandez, Roman

    2016-08-01

    The aim of the present study was to evaluate the effect of conditioned medium from human uterine cervical stem cells (CM-hUCESCs) in uveitis. To do that, uveitis was induced in rats after footpad injection of Escherichia coli lipopolysaccaride (LPS). Human retinal pigment epithelial (ARPE-19) cells after LPS challenge were used to test anti-inflammatory effect of CM-hUCESCs 'ìn vitro'. Real-time PCR was used to evaluate mRNA expression levels of the pro-inflammatory cytokines interkeukin-6, interkeukin-8, macrophage inflammatory protein-1 alpha, tumor necrosis factor alpha, and the anti-inflammatory interkeukin-10. Leucocytes from aqueous humor (AqH) were quantified in a Neubauer chamber, and eye histopathological analysis was done with hematoxylin-eosin staining. Additionally, using a human cytokine antibody array we evaluated CM-hUCESCs to determine mediating proteins. Results showed that administration of CM-hUCESCs significantly reduced LPS-induced pro-inflammatory cytokines both 'in vitro' and 'in vivo', and decreased leucocytes in AqH and ocular tissues. High levels of cytokines with anti-inflammatory effects were found in CM-hUCESCs, suggesting a possible role of these factors in reducing intraocular inflammation. In summary, treatment with CM-hUCESCs significantly reduces inflammation in uveitis. Our data indicate that CM-hUCESCs could be regarded as a potential therapeutic agent for patients suffering from ocular inflammation. Copyright © 2016 Elsevier Ltd. All rights reserved.

  18. [Lactobacillus rhamnosus GG conditioned medium prevents E. coli meningitis by inhibiting nuclear factor-κB pathway].

    PubMed

    Zeng, Qing; He, Xiao-Long; Xiao, Han-Sheng; DU, Lei; Li, Yu-Jing; Chen, Le-Cheng; Tian, Hui-Wen; Huang, Sheng-He; Cao, Hong

    2017-01-20

    To investigate whether Lactobacillus rhamnosus GG conditioned medium(LGG-CM)has preventive effect against E. coli K1-induced neuropathogenicity in vitro by inhibiting nuclear factor-κB (NF-κB) signaling pathway. An in vitro blood-brain barrier (BBB) model was constructed using human brain microvascular endothelial cells (HBMECs). The effect of LGG-CM on E. coli-actived NF-κB signaling pathway was assayed using Western blotting. Invasion assay and polymorphonuclear leukocyte (PMN) transmigration assay were performed to explore whether LGG-CM could inhibit E. coli invasion and PMN transmigration across the BBB in vitro. The expressions of ZO-1 and CD44 were detected using Western blotting and immunofluorescence. The changes of trans-epithelial electric resistance (TEER) and bacterial translocation were determined to evaluate the BBB permeability. Pre-treament with LGG-CM inhibited E. coli-activated NF-κB signaling pathway in HBMECs and decreased the invasion of E. coli K1 and transmigration of PMN. Western blotting showed that LGG-CM could alleviate E. coli-induced up-regulation of CD44 and down-regulation of ZO-1 expressions in HBMECs. In addition, pre-treatment with LGG-CM alleviated E. coli K1-induced reduction of TEER and suppressed bacterial translocation across the BBB in vitro. LGG-CM can block E. coli-induced activation of NF-κB signaling pathway and thereby prevents E. coli K1-induced neuropathogenicity by decreasing E. coli K1 invasion rates and PMN transmigration.

  19. Conditioned Medium from Periodontal Ligament Stem Cells Enhances Periodontal Regeneration.

    PubMed

    Nagata, Mizuki; Iwasaki, Kengo; Akazawa, Keiko; Komaki, Motohiro; Yokoyama, Naoki; Izumi, Yuichi; Morita, Ikuo

    2017-05-01

    Periodontal disease is one of the most common infectious diseases in adults and is characterized by the destruction of tooth-supporting tissues. Mesenchymal stem cells (MSCs) comprise the mesoderm-originating stem cell population, which has been studied and used for cell therapy. However, because of the lower rate of cell survival after MSC transplantation in various disease models, paracrine functions of MSCs have been receiving increased attention as a regenerative mechanism. The aim of this study was to investigate the regenerative potential of transplanted conditioned medium (CM) obtained from cultured periodontal ligament stem cells (PDLSCs), the adult stem cell population in tooth-supporting tissues, using a rat periodontal defect model. Cell-free CM was collected from PDLSCs and fibroblasts, using ultrafiltration and transplanted into surgically created periodontal defects. Protein content of CM was examined by antibody arrays. Formation of new periodontal tissues was analyzed using microcomputed tomography and histological sections. PDLSC-CM transplantation enhanced periodontal tissue regeneration in a concentration-dependent manner, whereas fibroblast-CM did not show any regenerative function. Proteomic analysis revealed that extracellular matrix proteins, enzymes, angiogenic factors, growth factors and cytokines were contained in PDLSC-CM. Furthermore, PDLSC-CM transplantation resulted in the decreased mRNA level of tumor necrosis factor-α (TNF-α) in healing periodontal tissues. In addition, we found that PDLSC-CM suppressed the mRNA level of TNF-α in the monocyte/macrophage cell line, RAW cells, stimulated with IFN-γ. Our findings suggested that PDLSC-CM enhanced periodontal regeneration by suppressing the inflammatory response through TNF-α production, and transplantation of PDLSC-CM could be a novel approach for periodontal regenerative therapy.

  20. Conditioned Medium from Periodontal Ligament Stem Cells Enhances Periodontal Regeneration

    PubMed Central

    Nagata, Mizuki; Akazawa, Keiko; Komaki, Motohiro; Yokoyama, Naoki; Izumi, Yuichi; Morita, Ikuo

    2017-01-01

    Periodontal disease is one of the most common infectious diseases in adults and is characterized by the destruction of tooth-supporting tissues. Mesenchymal stem cells (MSCs) comprise the mesoderm-originating stem cell population, which has been studied and used for cell therapy. However, because of the lower rate of cell survival after MSC transplantation in various disease models, paracrine functions of MSCs have been receiving increased attention as a regenerative mechanism. The aim of this study was to investigate the regenerative potential of transplanted conditioned medium (CM) obtained from cultured periodontal ligament stem cells (PDLSCs), the adult stem cell population in tooth-supporting tissues, using a rat periodontal defect model. Cell-free CM was collected from PDLSCs and fibroblasts, using ultrafiltration and transplanted into surgically created periodontal defects. Protein content of CM was examined by antibody arrays. Formation of new periodontal tissues was analyzed using microcomputed tomography and histological sections. PDLSC-CM transplantation enhanced periodontal tissue regeneration in a concentration-dependent manner, whereas fibroblast-CM did not show any regenerative function. Proteomic analysis revealed that extracellular matrix proteins, enzymes, angiogenic factors, growth factors and cytokines were contained in PDLSC-CM. Furthermore, PDLSC-CM transplantation resulted in the decreased mRNA level of tumor necrosis factor-α (TNF-α) in healing periodontal tissues. In addition, we found that PDLSC-CM suppressed the mRNA level of TNF-α in the monocyte/macrophage cell line, RAW cells, stimulated with IFN-γ. Our findings suggested that PDLSC-CM enhanced periodontal regeneration by suppressing the inflammatory response through TNF-α production, and transplantation of PDLSC-CM could be a novel approach for periodontal regenerative therapy. PMID:28027709

  1. Endothelial cell differentiation into capillary-like structures in response to tumour cell conditioned medium: a modified chemotaxis chamber assay.

    PubMed

    Garrido, T; Riese, H H; Aracil, M; Pérez-Aranda, A

    1995-04-01

    We have developed a modified chemotaxis chamber assay in which bovine aortic endothelial (BAE) cells degrade Matrigel basement membrane and migrate and form capillary-like structures on type I collagen. This capillary formation occurs in the presence of conditioned media from highly metastatic tumour cell lines, such as B16F10 murine melanoma or MDA-MD-231 human breast adenocarcinoma, but not in the presence of conditioned medium (CM) from the less invasive B16F0 cell line. Replacement of tumour cell CM by 10 ng ml-1 basic fibroblast growth factor (bFGF) also results in capillary-like structure formation by BAE cells. An anti-bFGF antibody blocks this effect, showing that bFGF is one of the factors responsible for the angiogenic response induced by B16F10 CM in our assay. Addition of an anti-laminin antibody reduces significantly the formation of capillary-like structures, probably by blocking the attachment of BAE cells to laminin present in Matrigel. The anti-angiogenic compound suramin inhibits in a dose-dependent manner (complete inhibition with 100 microM suramin) the migration and differentiation of BAE cells on type I collagen in response to B16F10 CM. This assay represents a new model system to study tumour-induced angiogenesis in vitro.

  2. Corneal epithelial wound healing and bactericidal effect of conditioned medium from human uterine cervical stem cells.

    PubMed

    Bermudez, Maria A; Sendon-Lago, Juan; Eiro, Noemi; Treviño, Mercedes; Gonzalez, Francisco; Yebra-Pimentel, Eva; Giraldez, Maria Jesus; Macia, Manuel; Lamelas, Maria Luz; Saa, Jorge; Vizoso, Francisco; Perez-Fernandez, Roman

    2015-01-22

    To evaluate the effect of conditioned medium from human uterine cervical stem cells (CM-hUCESCs) on corneal epithelial healing in a rat model of dry eye after alkaline corneal epithelial ulcer. We also tested the bactericidal effect of CM-hUCESCs. Dry eye was induced in rats by extraocular lacrimal gland excision, and corneal ulcers were produced using NaOH. Corneal histologic evaluation was made with hematoxylin-eosin (H&E) staining. Real-time PCR was used to evaluate mRNA expression levels of proinflammatory cytokines. We also studied the bactericidal effect of CM-hUCESCs in vitro and on infected corneal contact lenses (CLs) using Escherichia coli and Staphylococcus epidermidis bacteria. In addition, in order to investigate proteins from CM-hUCESCs that could mediate these effects, we carried out a human cytokine antibody array. After injury, dry eyes treated with CM-hUCESCs significantly improved epithelial regeneration and showed reduced corneal macrophage inflammatory protein-1 alpha (MIP-1α) and TNF-α mRNA expression as compared to untreated eyes and eyes treated with culture medium or sodium hyaluronate ophthalmic drops. In addition, we found in CM-hUCESCs high levels of proteins, such as tissue inhibitors of metalloproteinases 1 and 2, fibroblast growth factor 6 and 7, urokinase receptor, and hepatocyte growth factor, that could mediate these effects. In vitro, CM-hUCESCs showed a clear bactericidal effect on both E. coli and S. epidermidis and CLs infected with S. epidermidis. Analyses of CM-hUCESCs showed elevated levels of proteins that could be involved in the bactericidal effect, such as the chemokine (C-X-C motif) ligands 1, 6, 8, 10, and the chemokine (C-C motif) ligands 5 and 20. Treatment with CM-hUCESCs improved wound healing of alkali-injured corneas and showed a strong bactericidal effect on CLs. Patients using CLs and suffering from dry eye, allergies induced by commercial solutions, or small corneal injuries could benefit from this treatment. Copyright 2015 The Association for Research in Vision and Ophthalmology, Inc.

  3. Secretome of Differentiated PC12 Cells Enhances Neuronal Differentiation in Human Mesenchymal Stem Cells Via NGF-Like Mechanism.

    PubMed

    Srivastava, A; Singh, S; Pandey, A; Kumar, D; Rajpurohit, C S; Khanna, V K; Pant, A B

    2018-03-12

    The secretome-mediated responses over cellular physiology are well documented. Stem cells have been ruling the field of secretomics and its role in regenerative medicine since the past few years. However, the mechanistic aspects of secretome-mediated responses and the role of other cells in this area remain somewhat elusive. Here, we investigate the effects of secretome-enriched conditioned medium (CM) of neuronally differentiated PC12 cells on the neuronal differentiation of human mesenchymal stem cells (hMSCs). The exposure to CM at a ratio of 1:1 (CM: conditioned medium of PC12 cells) led to neuronal induction in hMSCs. This neuronal induction was compared with a parallel group of cells exposed to nerve growth factor (NGF). There was a marked increase in neurite length and expression of neuronal markers (β-III tubulin, neurofilament-M (NF-M), synaptophysin, NeuN in exposed hMSCs). Experimental group co-exposed to NGF and CM showed an additive response via MAPK signaling and directed the cells particularly towards cholinergic lineage. The ability of CM to enhance the neuronal properties of stem cells could aid in their rapid differentiation into neuronal subtypes in case of stem cell transplantation for neuronal injuries, thus broadening the scope of non-stem cell-based applications in the area of secretomics.

  4. Simulation of cosmic irradiation conditions in thick target arrangements

    NASA Technical Reports Server (NTRS)

    Theis, S.; Englert, P.; Reedy, R. C.; Arnold, J. R.

    1986-01-01

    One approach to simulate 2-pi irradiation conditions of planetary surfaces which has been widely applied in the past are bombardments of so called thick targets. A very large thick target was exposed recently to 2.1 GeV protons at the Bevatron-Bevalac in Berkeley. In a 100x100x180 cm steel-surrounded granodiorite target radioactive medium and high energy spallation products of the incident primary and of secondary particles were analyzed along the beam axis down to depths of 140 g/cm(2) in targets such as Cu, Ni, Co, Fe, T, Si, SiO2 and Al. Activities of these nuclides were exclusively determined via instrumental gamma-ray spectroscopy. Relative yields of neutron capture and spallation products induced in Co and Cu targets during the thick target bombardment are shown as a function of depth. The majority of the medium energy products such as Co-58 from Co targets exhibit a maximum at shallow depths of 40-60 g/cm(2) and then decrease exponentially. In a comparable 600 MeV proton bombarded thick target such a slight maximum for medium energy products was not observed. Rather, Co-58 activities in Co decreased steadily with the highest activity at the surface. The activities of the n-capture product Co-60 increase steadily starting at the surface. This indicates the rapidly growing flux of low energy neutrons within the target.

  5. Feeder & basic fibroblast growth factor-free culture of human embryonic stem cells: Role of conditioned medium from immortalized human feeders.

    PubMed

    Teotia, Pooja; Sharma, Shilpa; Airan, Balram; Mohanty, Sujata

    2016-12-01

    Human embryonic stem cell (hESC) lines are commonly maintained on inactivated feeder cells, in the medium supplemented with basic fibroblast growth factor (bFGF). However, limited availability of feeder cells in culture, and the high cost of growth factors limit their use in scalable expansion of hESC cultures for clinical application. Here, we describe an efficient and cost-effective feeder and bFGF-free culture of hESCs using conditioned medium (CM) from immortalized feeder cells. KIND-1 hESC cell line was cultured in CM, collected from primary mouse embryonic fibroblast, human foreskin fibroblast (HFF) and immortalized HFF (I-HFF). Pluripotency of KIND-1 hESC cell line was confirmed by expression of genes, proteins and cell surface markers. In culture, these cells retained normal morphology, expressed all cell surface markers, could differentiate to embryoid bodies upon culture in vitro. Furthermore, I-HFF feeder cells without supplementation of bFGF released ample amount of endogenous bFGF to maintain stemness of hESC cells. The study results described the use of CM from immortalized feeder cells as a consistent source and an efficient, inexpensive feeder-free culture system for the maintenance of hESCs. Moreover, it was possible to maintain hESCs without exogenous supplementation of bFGF. Thus, the study could be extended to scalable expansion of hESC cultures for therapeutic purposes.

  6. Electrochemical Oxidation of Resorcinol in Aqueous Medium Using Boron-Doped Diamond Anode: Reaction Kinetics and Process Optimization with Response Surface Methodology

    PubMed Central

    Körbahti, Bahadır K.; Demirbüken, Pelin

    2017-01-01

    Electrochemical oxidation of resorcinol in aqueous medium using boron-doped diamond anode (BDD) was investigated in a batch electrochemical reactor in the presence of Na2SO4 supporting electrolyte. The effect of process parameters such as resorcinol concentration (100–500 g/L), current density (2–10 mA/cm2), Na2SO4 concentration (0–20 g/L), and reaction temperature (25–45°C) was analyzed on electrochemical oxidation using response surface methodology (RSM). The optimum operating conditions were determined as 300 mg/L resorcinol concentration, 8 mA/cm2 current density, 12 g/L Na2SO4 concentration, and 34°C reaction temperature. One hundred percent of resorcinol removal and 89% COD removal were obtained in 120 min reaction time at response surface optimized conditions. These results confirmed that the electrochemical mineralization of resorcinol was successfully accomplished using BDD anode depending on the process conditions, however the formation of intermediates and by-products were further oxidized at much lower rate. The reaction kinetics were evaluated at optimum conditions and the reaction order of electrochemical oxidation of resorcinol in aqueous medium using BDD anode was determined as 1 based on COD concentration with the activation energy of 5.32 kJ/mol that was supported a diffusion-controlled reaction. PMID:29082225

  7. Stromal Fibroblasts from the Interface Zone of Triple Negative Breast Carcinomas Induced Epithelial-Mesenchymal Transition and its Inhibition by Emodin

    PubMed Central

    Wang, Hao-Yu; Hung, Chao-Ming; Lin, Ying-Chao; Ho, Chi-Tang; Way, Tzong-Der

    2017-01-01

    “Triple negative breast cancer” (TNBC) is associated with a higher rate and earlier time of recurrence and worse prognosis after recurrence. In this study, we aimed to examine the crosstalk between fibroblasts and TNBC cells. The fibroblasts were isolated from TNBC patients’ tissue in tumor burden zones, distal normal zones and interface zones. The fibroblasts were indicated as cancer-associated fibroblasts (CAFs), normal zone fibroblasts (NFs) and interface zone fibroblasts (INFs). Our study found that INFs grew significantly faster than NFs and CAFs in vitro. The epithelial BT20 cells cultured with the conditioned medium of INFs (INFs-CM) and CAFs (CAFs-CM) showed more spindle-like shape and cell scattering than cultured with the conditioned medium of NFs (NFs-CM). These results indicated that factors secreted by INFs-CM or CAFs-CM could induce the epithelial-mesenchymal transition (EMT) phenotype in BT20 cells. Using an in vitro co-culture model, INFs or CAFs induced EMT and promoted cancer cell migration in BT20 cells. Interestingly, we found that emodin inhibited INFs-CM or CAFs-CM-induced EMT programming and phenotype in BT20 cells. Previous studies reported that CAFs and INFs-secreted TGF-β promoted human breast cancer cell proliferation, here; our results indicated that TGF-β initiated EMT in BT20 cells. Pretreatment with emodin significantly suppressed the TGF-β-induced EMT and cell migration in BT20 cells. These results suggest that emodin may be used as a novel agent for the treatment of TNBC. PMID:28060811

  8. Hair-growth stimulation by conditioned medium from vitamin D3-activated preadipocytes in C57BL/6 mice.

    PubMed

    Jung, Min Kyung; Ha, Soogyeong; Huh, Scarlett Yoona; Park, Seung Beom; Kim, Sangyoon; Yang, Yoolhee; Kim, Daejin; Hur, Dae Young; Jeong, Hyuk; Bang, Sa Ik; Park, Hyunjeong; Cho, Daeho

    2015-05-01

    Recently, immature adipocyte lineage cells have been suggested as a potential hair-growth stimulator. Diverse studies have been attempted to find methods for the preconditioning of immature adipocyte lineage cells. The present study investigates the effect of conditioned medium (CM) from vitamin D3 (Vd3) pre-activated preadipocytes on hair-growth ability. To test the effect of CM from Vd3 pre-activated preadipocytes on hair-growth efficiency in mice, we compared the differences in hair regenerated after injecting CM from mouse preadipocytes pre-activated with or without Vd3. Next, to determine the regulating factors, the VEGF level was measured by ELISA and angiogenesis level was evaluated by IHC. Finally, the signaling mechanism was investigated by inhibitor kinase assay and western blotting. The CM from Vd3 pre-activated preadipocyte injection markedly promoted the ability of hair regeneration in mice. The VEGF levels were increased by Vd3 treatment in vitro and the CM from Vd3 pre-activated preadipocytes significantly increased the angiogenesis in vivo, suggesting the involvement of angiognensis in the hair regeneration induced by CM from pre-activated preadipocytes. In signaling study, Vd3-enhanced VEGF production was reduced by an ERK1/2 inhibitor and the level of ERK1/2 phosphorylation was increased by treatment with Vd3. This has been the first report on CM from Vd3 pre-activated preadipocyte displaying stimulatory effects on hair growth via the enhancement of angiogenesis in a hairless-induced C57BL/6 mice. Copyright © 2015 Elsevier Inc. All rights reserved.

  9. Conditioned Medium Derived from Neural Progenitor Cells Induces Long-term Post-ischemic Neuroprotection, Sustained Neurological Recovery, Neurogenesis, and Angiogenesis.

    PubMed

    Doeppner, Thorsten R; Traut, Viktorija; Heidenreich, Alexander; Kaltwasser, Britta; Bosche, Bert; Bähr, Mathias; Hermann, Dirk M

    2017-03-01

    Adult neural progenitor cells (NPCs) induce post-ischemic long-term neuroprotection and brain remodeling by releasing of survival- and plasticity-promoting mediators. To evaluate whether secreted factors may mimic neuroprotective and restorative effects of NPCs, we exposed male C57BL6 mice to focal cerebral ischemia and intravenously applied conditioned medium (CM) derived from subventricular zone NPCs. CM dose-dependently reduced infarct volume and brain leukocyte infiltration after 48 h when delivered up to 12 h after focal cerebral ischemia. Neuroprotection persisted in the post-acute stroke phase yielding enhanced neurological recovery that lasted throughout the 28-day observation period. Increased Bcl-2, phosphorylated Akt and phosphorylated STAT-3 abundance, and reduced caspase-3 activity and Bax abundance were noted in ischemic brains of CM-treated mice at 48 h post-stroke, indicative of enhanced cell survival signaling. Long-term neuroprotection was associated with increased brain glial cell line-derived neurotrophic factor (GDNF) and vascular endothelial growth factor (VEGF) concentrations at 28 days resulting in increased neurogenesis and angiogenesis. The observation that NPC-derived CM induces sustained neuroprotection and neurological recovery suggests that cell transplantation may be dispensable when secreted factors are instead administered.

  10. Therapy with mesenchymal stromal cells or conditioned medium reverse cardiac alterations in a high-fat diet-induced obesity model.

    PubMed

    Daltro, P S; Barreto, B C; Silva, P G; Neto, P Chenaud; Sousa Filho, P H F; Santana Neta, D; Carvalho, G B; Silva, D N; Paredes, B D; de Alcantara, A C; Freitas, L A R; Couto, R D; Santos, R R; Souza, B S F; Soares, M B P; Macambira, S G

    2017-10-01

    Obesity is associated with numerous cardiac complications, including arrhythmias, cardiac fibrosis, remodeling and heart failure. Here we evaluated the therapeutic potential of mesenchymal stromal cells (MSCs) and their conditioned medium (CM) to treat cardiac complications in a mouse model of high-fat diet (HFD)-induced obesity. After obesity induction and HFD withdrawal, obese mice were treated with MSCs, CM or vehicle. Cardiac function was assessed using electrocardiography, echocardiography and treadmill test. Body weight and biochemical parameters were evaluated. Cardiac tissue was used for real time (RT)-polymerase chain reaction (PCR) and histopathologic analysis. Characterization of CM by protein array showed the presence of different cytokines and growth factors, including chemokines, osteopontin, cystatin C, Serpin E1 and Gas 6. HFD-fed mice presented cardiac arrhythmias, altered cardiac gene expression and fibrosis reflected in physical exercise incapacity associated with obesity and diabetes. Administration of MSCs or CM improved arrhythmias and exercise capacity. This functional improvement correlated with normalization of GATA4 gene expression in the hearts of MSC- or CM-treated mice. The gene expression of connexin 43, troponin I, adiponectin, transforming growth factor (TGF) β, peroxisome proliferator activated receptor gamma (PPARγ), insulin-like growth factor 1 (IGF-1), matrix metalloproteinase-9 (MMP9) and tissue inhibitor of metalloproteinases 1 (TIMP1) were significantly reduced in MSCs, but not in CM-treated mice. Moreover, MSC or CM administration reduced the intensity of cardiac fibrosis. Our results suggest that MSCs and CM have a recovery effect on cardiac disturbances due to obesity and corroborate to the paracrine action of MSCs in heart disease models. Copyright © 2017 International Society for Cellular Therapy. Published by Elsevier Inc. All rights reserved.

  11. Human umbilical cord stem cell conditioned medium versus serum-free culture medium in the treatment of cryopreserved human ovarian tissues in in-vitro culture: a randomized controlled trial.

    PubMed

    Jia, Yingxian; Shi, Xiaohan; Xie, Yidong; Xie, Xiaochuan; Wang, Yan; Li, Shangwei

    2017-06-24

    To reduce young female fertility loss, the in-vitro culture of cryopreserved ovarian cortical tissues (OCTs) is considered an effective approach without delaying treatment and undergoing stimulation medicine. However, ischemic damage and follicular loss during the in-vitro culture of OCTs are major technical challenges. Human umbilical cord stem cells (HUMSCs) and their conditioned medium (HUMSC-CM) have been considered to be potential resources for regeneration medicine because they secrete cytokines and enhance cell survival and function. The aim of this study was to determine whether HUMSC-CM improves the development of frozen-thawed in-vitro cultured ovarian tissues compared with a serum-free culture medium (SF-CM). The thawed OCTs (n = 68) were cultivated in HUMSC-CM and SF-CM in vitro for 8 days, and the ovarian tissues were processed and analyzed by a classical histological evaluation. The microvessel density (MVD) and apotosis detection during in-vitro culture of OCTs were also performed. A significant difference in the rate of morphologically normal primordial follicles in the HUMSC-CM group was observed compared to that in the SF-CM group (group C) from days 2 to 4 (day 2: group B 58.0 ± 2.45% vs group C 32.0 ± 5.83%, p = 0.002; day 3: group B 55.5 ± 4.20% vs group C 21.0 ± 9.80%, p = 0.048; day 4: group B 52.0 ± 4.08% vs group C 21.5 ± 8.19%, p = 0.019). The microvessel density (MVD) detection showed a time-dependent increase and peaked on day 4. There was a significant difference between groups B (49.33 ± 0.58) and C (24.33 ± 3.79) (p = 0.036). The percentage of apoptotic follicles in group B was lower than that in group C on day 1 (13.75 ± 2.50% vs 27.0 ± 10.10%, p = 0.003), day 5 (11.75 ± 1.50% vs 51.0 ± 10.5%, p = 0.019) and day 7 (15.0 ± 5.10% vs 46.5 ± 21.75%, p = 0.018). These data have provided the first experimental evidence of the effect of HUMSC-CM on frozen-thawed OCTs in vitro. The results showed that the HUMSC-CM group provided a better protecting effect on the in-vitro culture of the cryopreserved OCTs compared to the SF-CM group.

  12. Micropropagation, Micromorphological Studies, and In Vitro Flowering in Rungia pectinata L.

    PubMed

    Shekhawat, Mahipal S; Manokari, M; Ravindran, C P

    2016-01-01

    A tissue culture protocol was developed for an important medicinal plant Rungia pectinata L. in the present study. Nodal shoots were used as explants and surface-sterilized with 0.1% HgCl2 solution. Murashige and Skoog (MS) medium was used to establish the cultures of R. pectinata. The bud break was reported on MS medium supplemented with 1.0 mg L(-1) 6-benzylaminopurine (BAP). About 98% response was observed with this media combination and maximum 3.2 shoots per explant with 4.3 cm length were recorded. The shoots were further multiplied using MS medium augmented with 0.5 mg L(-1) each of BAP and kinetin (Kin) + 0.1 mg L(-1) indole-3 acetic acid (IAA). Maximum 13.2 shoots per explant with 5.2 cm length were observed. All the shoots were rooted (4.9 roots per shoot with 3.5 cm length) on half strength MS medium fortified with 2.0 mg L(-1) indole-3 butyric acid (IBA). In vitro flowering was induced from the shoots on half strength MS medium supplemented with same concentrations and combinations of growth regulators used for shoot multiplication under 12/12 hr light/dark photoperiod. The plantlets were hardened in the greenhouse for two months and finally transferred to the field. The foliar micromorphological studies revealed the developmental changes in stomata, vein density, and trichomes during the culture of shoots under in vitro conditions.

  13. Micropropagation, Micromorphological Studies, and In Vitro Flowering in Rungia pectinata L.

    PubMed Central

    Shekhawat, Mahipal S.; Manokari, M.; Ravindran, C. P.

    2016-01-01

    A tissue culture protocol was developed for an important medicinal plant Rungia pectinata L. in the present study. Nodal shoots were used as explants and surface-sterilized with 0.1% HgCl2 solution. Murashige and Skoog (MS) medium was used to establish the cultures of R. pectinata. The bud break was reported on MS medium supplemented with 1.0 mg L−1 6-benzylaminopurine (BAP). About 98% response was observed with this media combination and maximum 3.2 shoots per explant with 4.3 cm length were recorded. The shoots were further multiplied using MS medium augmented with 0.5 mg L−1 each of BAP and kinetin (Kin) + 0.1 mg L−1 indole-3 acetic acid (IAA). Maximum 13.2 shoots per explant with 5.2 cm length were observed. All the shoots were rooted (4.9 roots per shoot with 3.5 cm length) on half strength MS medium fortified with 2.0 mg L−1 indole-3 butyric acid (IBA). In vitro flowering was induced from the shoots on half strength MS medium supplemented with same concentrations and combinations of growth regulators used for shoot multiplication under 12/12 hr light/dark photoperiod. The plantlets were hardened in the greenhouse for two months and finally transferred to the field. The foliar micromorphological studies revealed the developmental changes in stomata, vein density, and trichomes during the culture of shoots under in vitro conditions. PMID:27242948

  14. In vitro transdentinal effect of low-level laser therapy

    NASA Astrophysics Data System (ADS)

    Oliveira, C. F.; Basso, F. G.; dos Reis, R. I.; Parreiras-e-Silva, L. T.; Lins, E. C.; Kurachi, C.; Hebling, J.; Bagnato, V. S.; de Souza Costa, C. A.

    2013-05-01

    Low-level laser therapy (LLLT) has been used for the treatment of dentinal hypersensitivity. However, the specific LLL dose and the response mechanisms of these cells to transdentinal irradiation have not yet been demonstrated. Therefore, this study evaluated the transdentinal effects of different LLL doses on stressed odontoblast-like pulp cells MDPC-23 seeded onto the pulpal side of dentin discs obtained from human third molars. The discs were placed in devices simulating in vitro pulp chambers and the whole set was placed in 24-well plates containing plain culture medium (DMEM). After 24 h incubation, the culture medium was replaced by fresh DMEM supplemented with either 5% (simulating a nutritional stress condition) or 10% fetal bovine serum (FBS). The cells were irradiated with doses of 15 and 25 J cm-2 every 24 h, totaling three applications over three consecutive days. The cells in the control groups were removed from the incubator for the same times as used in their respective experimental groups for irradiation, though without activating the laser source (sham irradiation). After 72 h of the last active or sham irradiation, the cells were evaluated with respect to succinic dehydrogenase (SDH) enzyme production (MTT assay), total protein (TP) expression, alkaline phosphatase (ALP) synthesis, reverse transcriptase polymerase chain reaction (RT-PCR) for collagen type 1 (Col-I) and ALP, and morphology (SEM). For both tests, significantly higher values were obtained for the 25 J cm-2 dose. Regarding SDH production, supplementation of the culture medium with 5% FBS provided better results. For TP and ALP expression, the 25 J cm-2 presented higher values, especially for the 5% FBS concentration (Mann-Whitney p < 0.05). Under the tested conditions, near infrared laser irradiation at 25 J cm-2 caused transdentinal biostimulation of odontoblast-like MDPC-23 cells.

  15. Human Umbilical Cord-Derived Mesenchymal Stem Cells Conditioned Medium Attenuate Interstitial Fibrosis and Stimulate the Repair of Tubular Epithelial Cells in an Irreversible Model of Unilateral Ureteral Obstruction.

    PubMed

    Liu, Bo; Ding, Feng-Xia; Liu, Yang; Xiong, Geng; Lin, Tao; He, Da-Wei; Zhang, Yuan-Yuan; Zhang, De-Ying; Wei, Guang-Hui

    2017-07-01

    The growing number of patients suffering from chronic renal disease (CKD) is a challenge for the development of innovative therapies. Researchers have studied the therapeutic effects of cell therapy in acute kidney injury (AKI). However, the therapeutical effect of conditional medium (CM) in the CKD models have been rarely reported. Here, we examined the effects of umbilical cord derived-mesenchymal stem cells (hUC-MSCs) CM on renal fibrosis in a rat model of unilateral ureteral obstruction (UUO). We randomly divided the animals into three groups: sham-operated, UUO, UUO + CM. CM was administered via the left renal artery after total ligation of the left ureter. Rats were killed after 14 days of obstruction. Histological changes and oxidative stress parameters were assessed. Western blotting and immunohistochemistry analysis were used to measure epithelial-mesenchymal transition (EMT) markers, including epithelial cadherin (E-cadherin), α-smooth muscle actin (α-SMA), tumor necrosis factor-α (TNF-α), Collagen-I, and transforming growth factor β1 (TGF-β1). Proliferation and apoptosis of renal tubular epithelial cells (RTEs) were also measured. HucMSC-CM significantly reduced the levels of malondialdehyde (MDA) and reactive oxygen species (ROS), and increased the activity of glutathione (GSH) induced by UUO. Moreover, CM significantly reduced the expression of TGF-β1, α-SMA, TNF-α and Collagen-I in UUO kidney, promoted the proliferation of RTEs and inhibited its apoptosis. In addition, the increased expression of E-cadherin also reflects the effective improvement of renal interstitial fibrosis. This study shows that CM protects UUO-induced kidney damage and therefore could be a potential tool to prevent CKD progression. This article is protected by copyright. All rights reserved.

  16. Notochordal cell conditioned medium stimulates mesenchymal stem cell differentiation toward a young nucleus pulposus phenotype

    PubMed Central

    2010-01-01

    Introduction Mesenchymal stem cells (MSCs) offer promise for intervertebral disc (IVD) repair and regeneration because they are easily isolated and expanded, and can differentiate into several mesenchymal tissues. Notochordal (NC) cells contribute to IVD development, incorporate into the nucleus pulposus (NP), and stimulate mature disc cells. However, there have been no studies investigating the effects of NC cells on adult stem cell differentiation. The premise of this study is that IVD regeneration is more similar to IVD development than to IVD maintenance, and we hypothesize that soluble factors from NC cells differentiate MSCs to a phenotype characteristic of nucleus pulposus (NP) cells during development. The eventual clinical goal would be to isolate or chemically/recombinantly produce the active agent to induce the therapeutic effects, and to use it as either an injectable therapy for early intervention on disc disease, or in developing appropriately pre-differentiated MSC cells in a tissue engineered NP construct. Methods Human MSCs from bone marrow were expanded and pelleted to form high-density cultures. MSC pellets were exposed to either control medium (CM), chondrogenic medium (CM with dexamethasone and transforming growth factor, (TGF)-β3) or notochordal cell conditioned medium (NCCM). NCCM was prepared from NC cells maintained in serum free medium for four days. After seven days culture, MSC pellets were analyzed for appearance, biochemical composition (glycosaminoglycans and DNA), and gene expression profile (sox-9, collagen types-II and III, laminin-β1 and TIMP1(tissue inhibitor of metalloproteinases-1)). Results Significantly higher glycosaminoglycan accumulation was seen in NCCM treated pellets than in CM or TGFβ groups. With NCCM treatment, increased gene expression of collagen III, and a trend of increasing expression of laminin-β1 and decreased expression of sox-9 and collagen II relative to the TGFβ group was observed. Conclusions Together, results suggest NCCM stimulates mesenchymal stem cell differentiation toward a potentially NP-like phenotype with some characteristics of the developing IVD. PMID:20565707

  17. Lysophosphatidic acid-induced ADAM12 expression mediates human adipose tissue-derived mesenchymal stem cell-stimulated tumor growth.

    PubMed

    Do, Eun Kyoung; Kim, Young Mi; Heo, Soon Chul; Kwon, Yang Woo; Shin, Sang Hun; Suh, Dong-Soo; Kim, Ki-Hyung; Yoon, Man-Soo; Kim, Jae Ho

    2012-11-01

    Lysophosphatidic acid (LPA) is involved in mesenchymal stem cell-stimulated tumor growth in vivo. However, the molecular mechanism by which mesenchymal stem cells promote tumorigenesis remains elusive. In the present study, we demonstrate that conditioned medium from A549 human lung adenocarcinoma cells (A549 CM) induced the expression of ADAM12, a disintegrin and metalloproteases family member, in human adipose tissue-derived mesenchymal stem cells (hASCs). A549 CM-stimulated ADAM12 expression was abrogated by pretreatment of hASCs with the LPA receptor 1 inhibitor Ki16425 or by small interfering RNA-mediated silencing of LPA receptor 1, suggesting a key role for the LPA-LPA receptor 1 signaling axis in A549 CM-stimulated ADAM12 expression. Silencing of ADAM12 expression using small interfering RNA or short hairpin RNA abrogated LPA-induced expression of both α-smooth muscle actin, a marker of carcinoma-associated fibroblasts, and ADAM12 in hASCs. Using a xenograft transplantation model of A549 cells, we demonstrated that silencing of ADAM12 inhibited the hASC-stimulated in vivo growth of A549 xenograft tumors and the differentiation of transplanted hASCs to α-smooth muscle actin-positive carcinoma-associated fibroblasts. LPA-conditioned medium from hASCs induced the adhesion of A549 cells and silencing of ADAM12 inhibited LPA-induced expression of extracellular matrix proteins, periostin and βig-h3, in hASCs and LPA-conditioned medium-stimulated adhesion of A549 cells. These results suggest a pivotal role for LPA-stimulated ADAM12 expression in tumor growth and the differentiation of hASCs to carcinoma-associated fibroblasts expressing α-smooth muscle actin, periostin, and βig-h3. Copyright © 2012 Elsevier Ltd. All rights reserved.

  18. Multifaceted Therapeutic Benefits of Factors Derived From Dental Pulp Stem Cells for Mouse Liver Fibrosis.

    PubMed

    Hirata, Marina; Ishigami, Masatoshi; Matsushita, Yoshihiro; Ito, Takanori; Hattori, Hisashi; Hibi, Hideharu; Goto, Hidemi; Ueda, Minoru; Yamamoto, Akihito

    2016-10-01

    : Chronic liver injury from various causes often results in liver fibrosis (LF). Although the liver possesses endogenous tissue-repairing activities, these can be overcome by sustained inflammation and excessive fibrotic scar formation. Advanced LF leads to irreversible cirrhosis and subsequent liver failure and/or hepatic cancer. Here, using the mouse carbon tetrachloride (CCl 4 )-induced LF model, we showed that a single intravenous administration of stem cells derived from human exfoliated deciduous teeth (SHEDs) or of SHED-derived serum-free conditioned medium (SHED-CM) resulted in fibrotic scar resolution. SHED-CM suppressed the gene expression of proinflammatory mediators, such as TNF-α, IL-1β, and iNOS, and eliminated activated hepatic stellate cells by inducing their apoptosis, but protected parenchymal hepatocytes from undergoing apoptosis. In addition, SHED-CM induced tissue-repairing macrophages that expressed high levels of the profibrinolytic factor, matrix metalloproteinase 13. Furthermore, SHED-CM suppressed the CCl 4 -induced apoptosis of primary cultured hepatocytes. SHED-CM contained a high level of hepatocyte growth factor (HGF). Notably, HGF-depleted SHED-CM (dHGF-CM) did not suppress the proinflammatory response or resolve fibrotic scarring. Furthermore, SHED-CM, but not dHGF-CM, inhibited CCl 4 -induced hepatocyte apoptosis. These results suggest that HGF plays a central role in the SHED-CM-mediated resolution of LF. Taken together, our findings suggest that SHED-CM provides multifaceted therapeutic benefits for the treatment of LF. This study demonstrated that a single intravenous administration of stem cells from human exfoliated deciduous teeth (SHEDs) or of the serum-free conditioned medium (CM) derived from SHEDs markedly improved mouse liver fibrosis (LF). SHED-CM suppressed chronic inflammation, eliminated activated hepatic stellate cells by inducing their apoptosis, protected hepatocytes from undergoing apoptosis, and induced differentiation of tissue-repairing macrophages expressing high levels of the profibrinolytic factor matrix metalloproteinase 13. Furthermore, hepatocyte growth factor played a central role in the SHED-CM-mediated resolution of LF. This is the first report demonstrating the multifaceted therapeutic benefits of secreted factors derived from SHEDs for LF. ©AlphaMed Press.

  19. Mesenchymal stem cell-conditioned medium accelerates skin wound healing: an in vitro study of fibroblast and keratinocyte scratch assays.

    PubMed

    Walter, M N M; Wright, K T; Fuller, H R; MacNeil, S; Johnson, W E B

    2010-04-15

    We have used in vitro scratch assays to examine the relative contribution of dermal fibroblasts and keratinocytes in the wound repair process and to test the influence of mesenchymal stem cell (MSC) secreted factors on both skin cell types. Scratch assays were established using single cell and co-cultures of L929 fibroblasts and HaCaT keratinocytes, with wound closure monitored via time-lapse microscopy. Both in serum supplemented and serum free conditions, wound closure was faster in L929 fibroblast than HaCaT keratinocyte scratch assays, and in co-culture the L929 fibroblasts lead the way in closing the scratches. MSC-CM generated under serum free conditions significantly enhanced the wound closure rate of both skin cell types separately and in co-culture, whereas conditioned medium from L929 or HaCaT cultures had no significant effect. This enhancement of wound closure in the presence of MSC-CM was due to accelerated cell migration rather than increased cell proliferation. A number of wound healing mediators were identified in MSC-CM, including TGF-beta1, the chemokines IL-6, IL-8, MCP-1 and RANTES, and collagen type I, fibronectin, SPARC and IGFBP-7. This study suggests that the trophic activity of MSC may play a role in skin wound closure by affecting both dermal fibroblast and keratinocyte migration, along with a contribution to the formation of extracellular matrix. Copyright 2010 Elsevier Inc. All rights reserved.

  20. Comparison of fibroblast cell regeneration in three different concentrations of Wharton’s Jelly mesenchymal stem cells conditioned medium (WJMSCs-CM)

    NASA Astrophysics Data System (ADS)

    Untoro, E. G.; Asrianti, D.; Usman, M.; Meidyawati, R.; Margono, A.

    2017-08-01

    Wharton’s Jelly-derived mesenchymal stem cells (WJMSCs) have gained interest as an alternative source of stem cells for regenerative medicine. Although many studies have characterized Wharton’s Jelly biologically, the effects of different concentrations in a cultured medium have not yet been compared. Damaged fibroblasts, the primary components of irreversible dental pulpitis, irreversibly impair the ability to regenerate and lead to the disruption of extracellular matrix. This study was performed to evaluate the potency of three WJMSCs-CM concentrations in improving serum-starved fibroblasts. Fibroblasts were cultivated in five passages, and divided into four groups. The first group (the control group) consisted of fibroblast cells that had been treated using starvation methods. The other groups (the treatment groups) were treated with various concentration of WJMSCs-CM (50%, 25% and 12.5%). Proliferative ability was evaluated using a cell count method and analyzed with a one-way ANOVA. Cultivation of serum-starved fibroblasts produced significantly higher cell counts in 12.5% WJMSCs-CM compared to the 50% group. It can be concluded that 12.5% WJMSCs-CM is the most efficient concentration for fibroblast proliferation.

  1. HS 1603+3820 and its Warm Absorber

    NASA Astrophysics Data System (ADS)

    Nikołajuk, M.; Różańska, A.; Czerny, B.; Dobrzycki, A.

    2009-07-01

    We use photoionization codes CLOUDY and TITAN to obtain physical conditions in the absorbing medium close to the nucleus of a distant quasar (z = 2.54) HS 1603+3820. We found that the total column density of this Warm Absorber is 2 x 1022 cm-2. Due to the softness of the quasars spectrum the modelling allowed us also to determine uniquely the volume hydrogen density of this warm gas (n = 1010 cm-3) which combined with the other quasar parameters leads to a distance determination to the Warm Absorber from the central source which is ~ 1.5 x 1016 cm.

  2. Mixed-species biofilm formation by lactic acid bacteria and rice wine yeasts.

    PubMed

    Kawarai, Taketo; Furukawa, Soichi; Ogihara, Hirokazu; Yamasaki, Makari

    2007-07-01

    We found that species combinations such as Lactobacillus casei subsp. rhamnosus IFO3831 and Saccharomyces cerevisiae Kyokai-10 can form a mixed-species biofilm in coculture. Moreover, the Kyokai-10 yeast strain can form a biofilm in monoculture in the presence of conditioned medium (CM) from L. casei IFO3831. The active substance(s) in bacterial CM is heat sensitive and has a molecular mass of between 3 and 5 kDa. In biofilms from cocultures or CM monocultures, yeast cells had a distinct morphology, with many hill-like protrusions on the cell surface.

  3. Conditioned medium from persistently RSV-infected macrophages alters transcriptional profile and inflammatory response of non-infected macrophages.

    PubMed

    Rivera-Toledo, Evelyn; Salido-Guadarrama, Iván; Rodríguez-Dorantes, Mauricio; Torres-González, Laura; Santiago-Olivares, Carlos; Gómez, Beatriz

    2017-02-15

    Cells susceptible to persistent viral infections undergo important changes in their biological functions as a consequence of the expression of viral gene products that are capable of altering the gene expression profile of the host cell. Previously, we reported that persistence of the RSV genome in a mouse macrophage cell line induces important alterations in cell homeostasis, including constitutive expression of IFN-β and other pro-inflammatory cytokines. Here, we postulated that changes in the homeostasis of non-infected macrophages could be induced by soluble factors secreted by persistently RSV- infected macrophages. To test this hypothesis, non-infected mouse macrophages were treated with conditioned medium (CM) collected from cultures of persistently RSV-infected macrophages. Total RNA was extracted and a microarray-based gene expression analysis was performed. Non-infected macrophages, treated under similar conditions with CM obtained from cultures of non-infected macrophages, were used as a control to establish differential gene expression between the two conditions. Results showed that CM from the persistently RSV-infected cultures altered expression of a total of 95 genes in non-infected macrophages, resulting in an antiviral gene-transcription profile along with inhibition of the inflammatory response, since some inflammatory genes were down-regulated, including Nlrp3 and Il-1 β, both related to the inflammasome pathway. However, down-regulation of Nlrp3 and Il-1 β was reversible upon acute RSV infection. Additionally, we observed that the inflammatory response, evaluated by secreted IL-1 β, a final product of the inflammasome activity, was enhanced during acute RSV infection in macrophages treated with CM from persistently RSV-infected cultures, compared to that in macrophages treated with the control CM. This suggests that soluble factors secreted during RSV persistence may induce an exacerbated inflammatory response in non-infected cells. Copyright © 2017 Elsevier B.V. All rights reserved.

  4. SU-E-T-791: Validation of a Determinant Based Photon Transport Solver in Dose Perturbed By Diverse Media

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

    Kuo, H; Tome, W; Yaparpalvi, R

    Purpose: To validate a determinant based photon transport solver in dose imparted within different transition zone between different medium. Methods: Thickness (.2cm,.5cm, 1cm, 3cm) from various materials (Air - density=0.0012g/cm3, Cork-0.19g/cm3, Lung-0.26g/cm3, Bone-1.85g/cm3, Aluminum (Al)-2.7g/cm3, Titanium (Ti)-4.42g/cm3, Iron (Fe)-8g/cm3) were sandwiched by 10cm solid water. 6MV were used to study the calculation difference between a superposition photon beam model (AAA) and the determinant based Boltzmann photon transport solver (XB) at the upstream (I) and downstream boarder (II) of the medium, within the medium (III), and at far distance downstream away from medium (IV). Calculation was validated with available thickness ofmore » Air, Cork, Lung, Al, Ti and Fe. Results are presented as the ratio of the dose at the point with medium perturbation to the same point dose without perturbation. Results: Zone I showed different backscatter enhancement from high-density materials within the 5mm of the upstream border. AAA showed no backscatter at all, XB showed good agreement beyond 1mm upstream (1.18 vs 1.14, 1.09 vs 1.10, and 1.04 vs 1.05 for Fe, Ti, and Fe, respectively). Zone II showed a re-buildup after exiting high-density medium and Air but no build up for density close to water in both of the measurement and XB. AAA yielded the opposite results in Zone II. XB and AAA showed in Zone III very different absorption in high density medium and the Air. XB and measurement had high concordance regarding photon attenuation in Zone IV. AAA showed less agreement especially when the medium was Air or Fe. Conclusion: XB compared well with measurement in regions 1mm away from the interface. Planning using XB should be beneficial for External Beam Planning in situations with large air cavity, very low lung density, compact bone, and any kind of metal implant.« less

  5. Nearfield of a piston source of ultrasound in an absorbing medium.

    PubMed

    Nyborg, W L; Steele, R B

    1985-11-01

    Approximate expressions are discussed which are applicable for acoustic quantities in the vicinity of a plane piston source of ultrasound which radiates into an absorbing medium. A particularly useful approach for nearfield calculations combines an expression valid near the axis with another, given by Pierce [Acoustics, An Introduction to Its Physical Principles and Applications (McGraw-Hill, New York, 1981), Chap. 5], which is valid elsewhere. This approach gives reasonable accuracy at relatively low computational cost. Computed plots are presented, showing spatial distributions of the square of the pressure amplitude. Most of the plots are for a source diameter of 1.2 cm, a frequency of 3 MHz, and an absorption coefficient of 0.15 Np/cm; these are representative of conditions for medical applications of ultrasound.

  6. Transplantation of Human Umbilical Cord Blood-Derived Mesenchymal Stem Cells or Their Conditioned Medium Prevents Bone Loss in Ovariectomized Nude Mice

    PubMed Central

    An, Jee Hyun; Park, Hyojung; Song, Jung Ah; Ki, Kyung Ho; Yang, Jae-Yeon; Choi, Hyung Jin; Cho, Sun Wook; Kim, Sang Wan; Kim, Seong Yeon; Yoo, Jeong Joon; Baek, Wook-Young; Kim, Jung-Eun; Choi, Soo Jin; Oh, Wonil

    2013-01-01

    Umbilical cord blood (UCB) has recently been recognized as a new source of mesenchymal stem cells (MSCs) for use in stem cell therapy. We studied the effects of systemic injection of human UCB-MSCs and their conditioned medium (CM) on ovariectomy (OVX)-induced bone loss in nude mice. Ten-week-old female nude mice were divided into six groups: Sham-operated mice treated with vehicle (Sham-Vehicle), OVX mice subjected to UCB-MSCs (OVX-MSC), or human dermal fibroblast (OVX-DFB) transplantation, OVX mice treated with UCB-MSC CM (OVX-CM), zoledronate (OVX-Zol), or vehicle (OVX-Vehicle). Although the OVX-Vehicle group exhibited significantly less bone mineral density (BMD) gain compared with the Sham-Vehicle group, transplantation of hUCB-MSCs (OVX-MSC group) has effectively prevented OVX-induced bone mass attenuation. Notably, the OVX-CM group also showed BMD preservation comparable to the OVX-MSC group. In addition, microcomputed tomography analysis demonstrated improved trabecular parameters in both the OVX-MSC and OVX-CM groups compared to the OVX-Vehicle or OVX-DFB group. Histomorphometric analysis showed increased bone formation parameters, accompanied by increased serum procollagen type-I N-telopeptide levels in OVX-MSC and OVX-CM mice. However, cell-trafficking analysis failed to demonstrate engraftment of MSCs in bone tissue 48 h after cell infusion. In vitro, hUCB-MSC CM increased alkaline phosphatase (ALP) activity in human bone marrow-derived MSCs and mRNA expression of collagen type 1, Runx2, osterix, and ALP in C3H10T1/2 cells. Furthermore, hUCB-MSC CM significantly increased survival of osteocyte-like MLO-Y4 cells, while it inhibited osteoclastic differentiation. To summarize, transplantation of hUCB-MSCs could effectively prevent OVX-mediated bone loss in nude mice, which appears to be mediated by a paracrine mechanism rather than direct engraftment of the MSCs. PMID:23215868

  7. Morphological and Functional Attenuation of Degeneration of Peripheral Neurons by Mesenchymal Stem Cell-Conditioned Medium in Spinocerebellar Ataxia Type 1-Knock-in Mice.

    PubMed

    Suto, Nana; Mieda, Tokue; Iizuka, Akira; Nakamura, Kazuhiro; Hirai, Hirokazu

    2016-08-01

    Spinocerebellar ataxia type 1 (SCA1) is caused by the ataxin-1 protein (ATXN1) with an abnormally expanded polyglutamine tract and is characterized by progressive neurodegeneration. We previously showed that intrathecal injection of mesenchymal stem cells (MSCs) during the nonsymptomatic stage mitigates the degeneration of the peripheral nervous system (PNS) neurons in SCA1-knock-in (SCA1-KI) mice. We tested in this study whether the therapeutic effects of MSCs in SCA1-KI mice could be reproduced with MSC-releasing factor(s). To test the effects of MSC-releasing factor(s), we used MSC-conditioned medium (MSC-CM). MSC-CM was intrathecally and/or intravenously injected into young SCA1-KI mice, and the therapeutic effects were assessed in the PNS at later ages using immunostaining, electrophysiology, and behavioral tests. MSC-CM attenuated the degeneration of axons and myelin of spinal motor neurons. Consequently, the injected SCA1-KI mice exhibited smaller reductions in nerve conduction velocity in spinal motor neurons and reduced motor incoordination than the untreated mice. These results suggest that factors released from MSC mitigate the morphological and functional abnormalities in the PNS that are observed in SCA1-KI mice in a paracrine manner. © 2016 John Wiley & Sons Ltd.

  8. Pro-Inflammatory cytokines increases hepatocellular carcinoma cells thermotolerance: Evidence of how local inflammation may negatively impact radiofrequency ablation local control rates

    PubMed Central

    Douglas, Wade G.; Wang, Yangping; Gibbs, John F.; Tracy, Erin; Kuvshinoff, Boris; Huntoon, Kristin; Baumann, Heinz

    2008-01-01

    Background Hepatocellular carcinomas (HCC) associated with inflammation that undergo radiofrequency ablation (RFA) appear to have poorer local control rates. Little is known of how mediators of inflammation influence HCC cellular thermotolerance which in part is mediated by heat shock protein 70 (HSP 70). This study determines how inflammatory mediators effect cellular thermotolerance and provides insight into how associated inflammation may impact HCC RFA local control rates. Methods HepG2 cell lines were cultured in control medium (CM) or CM containing conditioned medium of endotoxin-activated macrophage (CMM). Serial dilutions of CMM established microenvironments approximating low, medium and high CMM. All groups underwent a heat shock challenge (HSC) at 45° C for 10 minutes. Western blot, northern blot, densometric analysis, along with Thymidine and clonagenic assays determined how inflammation influenced multiple biologic endpoints. Results Cells cultured in low CMM, expressed significantly more HSP 70 RNA and protein compared to control cells after HSC. The cells also had a higher proliferative and survival rate after HSC compared to control cells. Medium CMM cultured cells had no significant difference in HSP 70 RNA and protein production or proliferation and survival rates after HSC, compared to CM cultured cells. AT high CMM the inhibitory effects of inflammatory mediators prevailed, all the measured endpoints were significantly less compared to CM cultured cells. Conclusions This study demonstrates that inflammation can alter the responsiveness of HCC cells to a HSC in a dose dependent manner. This study supports the clinical observation that HCC associated with chronic inflammation have worse RFA local control rates. PMID:18262552

  9. Biomimetic evaluation of β tricalcium phosphate prepared by hot isostatic pressing

    PubMed Central

    Mateescu, Mihaela; Rguitti, Emmanuelle; Ponche, Arnaud; Descamps, Michel; Anselme, Karine

    2012-01-01

    Two types of completely densified β-TCP tablets were synthesized from a stoichiometric β-TCP powder. The first ones (TCP) were conventionally sintered, while the second ones (TCP-T) were sintered and treated by hot isostatic process (HIP). The HIP produced completely densified materials with relative densities greater than 99.9% and a transparent appearance of tablets. Samples were immersed in culture medium with (CM) or without serum (NCM) in static and dynamic conditions for a biomimetic evaluation. Similarly, SaOs-2 cells were cultured on samples in a static or dynamic flow perfusion system. The results of surface transformation in absence of cells showed that the dynamic condition increased the speed of calcium phosphate precipitations compared with the static condition. The morphology of precipitates was different with nature of tablets. The immersion in CM did impede this precipitation. XPS analysis of TCP-T tablets showed the presence of hydroxyapatite (HA) precipitates after incubation in NCM while octacalcium phosphate (OCP) precipitates were formed after incubation in CM. The analysis of the response of SaOs-2 cells on surfaces showed that the two types of materials are biocompatible. However, the dynamic mode of culture stimulated the differentiation of cells. Finally, it appears that the HIP treatment of TCP produces highly densified and transparent samples that display a good in vitro biocompatibility in static and dynamic culture conditions. Moreover, an interesting result of this work is the relationship between the presence of proteins in the immersion medium and the quality of precipitates formed on hipped TCP surface. PMID:23507861

  10. Mesenchymal stem cell-conditioned medium accelerates skin wound healing: An in vitro study of fibroblast and keratinocyte scratch assays

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

    Walter, M.N.M.; School of Life and Health Science, Aston University, Aston Triangle, Birmingham, B4 7EJ; Wright, K.T.

    2010-04-15

    We have used in vitro scratch assays to examine the relative contribution of dermal fibroblasts and keratinocytes in the wound repair process and to test the influence of mesenchymal stem cell (MSC) secreted factors on both skin cell types. Scratch assays were established using single cell and co-cultures of L929 fibroblasts and HaCaT keratinocytes, with wound closure monitored via time-lapse microscopy. Both in serum supplemented and serum free conditions, wound closure was faster in L929 fibroblast than HaCaT keratinocyte scratch assays, and in co-culture the L929 fibroblasts lead the way in closing the scratches. MSC-CM generated under serum free conditionsmore » significantly enhanced the wound closure rate of both skin cell types separately and in co-culture, whereas conditioned medium from L929 or HaCaT cultures had no significant effect. This enhancement of wound closure in the presence of MSC-CM was due to accelerated cell migration rather than increased cell proliferation. A number of wound healing mediators were identified in MSC-CM, including TGF-{beta}1, the chemokines IL-6, IL-8, MCP-1 and RANTES, and collagen type I, fibronectin, SPARC and IGFBP-7. This study suggests that the trophic activity of MSC may play a role in skin wound closure by affecting both dermal fibroblast and keratinocyte migration, along with a contribution to the formation of extracellular matrix.« less

  11. Acceleration of diabetic wound healing with adipose-derived stem cells, endothelial-differentiated stem cells, and topical conditioned medium therapy in a swine model.

    PubMed

    Irons, Robin F; Cahill, Kevin W; Rattigan, Deviney A; Marcotte, Joseph H; Fromer, Marc W; Chang, Shaohua; Zhang, Ping; Behling, Eric M; Behling, Kathryn C; Caputo, Francis J

    2018-05-09

    The purpose of our study was to investigate the effect of adipose-derived stem cells (ASCs), endothelial-differentiated ASCs (EC/ASCs), and various conditioned media (CM) on wound healing in a diabetic swine model. We hypothesized that ASC-based therapies would accelerate wound healing. Diabetes was induced in four Yorkshire swine through intravenous injection of streptozotocin. ASCs were harvested from flank fat and cultured in either M199 or EGM-2 medium. A duplicate series of seven full-thickness dorsal wounds were surgically created on each swine. The wounds in the cellular treatment group underwent injection of low-dose or high-dose ASCs or EC/ASCs on day 0, with a repeat injection of one half of the initial dose on day 15. Wounds assigned to the topical CM therapy were covered with 2 mL of either serum-free M199 primed by ASCs or human umbilical vein endothelial cells every 3 days. Wounds were assessed at day 0, 10, 15, 20, and 28. The swine were sacrificed on day 28. ImageJ software was used to evaluate the percentage of wound healing. The wounded skin underwent histologic, reverse transcription polymerase chain reaction, and enzyme-linked immunosorbent assay examinations to evaluate markers of angiogenesis and inflammation. We found an increase in the percentage of wound closure rates in cell-based treatments and topical therapies at various points compared with the untreated control wounds (P < .05). The results from the histologic, messenger RNA, and protein analyses suggested the treated wounds displayed increased angiogenesis and a diminished inflammatory response. Cellular therapy with ASCs, EC/ASCs, and topical CM accelerated diabetic wound healing in the swine model. Enhanced angiogenesis and immunomodulation might be key contributors to this process. The purpose of the present study was to translate the known beneficial effects of adipose-derived stem cells and associated conditioned medium therapy on diabetic wound healing to a large animal model. We demonstrated that stem cell and conditioned medium therapy significantly accelerate gross wound healing in diabetic swine, with data suggesting this might result from a decreased inflammatory response and increased angiogenesis. Copyright © 2018 Society for Vascular Surgery. Published by Elsevier Inc. All rights reserved.

  12. Carbofuran biodegradation in brackish groundwater and its effect on the hydraulic properties of the porous medium

    NASA Astrophysics Data System (ADS)

    Amiaz, Yanai; Ronen, Zeev; Adar, Eilon; Weisbrod, Noam

    2015-04-01

    A chalk fractured aquitard beneath an industrial site is subjected to intense contamination due to percolation of contaminants from the different facilities operating at the site. In order to reduce further contamination, draining trenches were excavated and filled with coarse gravel (3-4 cm in diameter) forming a porous medium, to which the contaminated groundwater discharges from the fractures surrounding the trenches. This research is aimed at establishing a biodegrading process of high efficiency and performance within the draining trenches. The research includes both field and laboratory experiments. An experimental setup of five columns (50 cm length and 4.5 cm in diameter) was constructed under highly controlled conditions. Over the course of the experiments, the columns were filled with different particle sizes and placed in a temperature controlled chamber. Filtered groundwater (0.2 µm) from the site groundwater, enriched by a model contaminant carbofuran (CRF), was injected to the columns; as two of the columns were inoculated by CRF degrading microorganisms native in the site's groundwater, two columns were inoculated by CRF degrading bacteria from the external environment, and one column was used as a control. During the experiment, measurements were taken from different locations along each column. These include: (a) CRF concentration and (b) hydraulic pressure and solution viscosity (in order to obtain the changes in permeability). A tracer test using uranine was carried out in parallel, in order to obtain the changes in hydraulic parameters. Correlating CRF concentration variations to changes of hydraulic parameters enable the deduction due to the effect that biological activity (under different temperature regimes) has on the hydraulic properties of the porous medium and its effect on the process of contaminant groundwater bodies' remediation. Preliminary results suggest that although biodegradation occurs, microbial activity has minor effect on the hydraulic properties of the porous medium under the explored conditions.

  13. Exosomes of human placenta-derived mesenchymal stem cells stimulate angiogenesis.

    PubMed

    Komaki, Motohiro; Numata, Yuri; Morioka, Chikako; Honda, Izumi; Tooi, Masayuki; Yokoyama, Naoki; Ayame, Hirohito; Iwasaki, Kengo; Taki, Atsuko; Oshima, Noriko; Morita, Ikuo

    2017-10-03

    The therapeutic potential of mesenchymal stem cells (MSCs) may be attributed partly to humoral factors such as growth factors, cytokines, and chemokines. Human term placental tissue-derived MSCs (PlaMSCs), or conditioned medium left over from cultures of these cells, have been reported to enhance angiogenesis. Recently, the exosome, which can transport a diverse suite of macromolecules, has gained attention as a novel intercellular communication tool. However, the potential role of the exosome in PlaMSC therapeutic action is not well understood. The purpose of this study was to evaluate PlaMSC-derived exosome angiogenesis promotion in vitro and in vivo. MSCs were isolated from human term placental tissue by enzymatic digestion. Conditioned medium was collected after 48-h incubation in serum-free medium (PlaMSC-CM). Angiogenic factors present in PlaMSC-CM were screened by a growth factor array. Exosomes were prepared by ultracentrifugation of PlaMSC-CM, and confirmed by transmission electron microscopy, dynamic light scattering, and western blot analyses. The proangiogenic activity of PlaMSC-derived exosomes (PlaMSC-exo) was assessed using an endothelial tube formation assay, a cell migration assay, and reverse transcription-PCR analysis. The in-vivo angiogenic activity of PlaMSC-exo was evaluated using a murine auricle ischemic injury model. PlaMSC-CM contained both angiogenic and angiostatic factors, which enhanced endothelial tube formation. PlaMSC-exo were incorporated into endothelial cells; these exosomes stimulated both endothelial tube formation and migration, and enhanced angiogenesis-related gene expression. Laser Doppler blood flow analysis showed that PlaMSC-exo infusion also enhanced angiogenesis in an in-vivo murine auricle ischemic injury model. PlaMSC-exo enhanced angiogenesis in vitro and in vivo, suggesting that exosomes play a role in the proangiogenic activity of PlaMSCs. PlaMSC-exo may be a novel therapeutic approach for treating ischemic diseases.

  14. The neuroprotective effect of rat adipose tissue-derived mesenchymal stem cell-conditioned medium on cortical neurons using an in vitro model of SCI inflammation.

    PubMed

    Szekiova, Eva; Slovinska, Lucia; Blasko, Juraj; Plsikova, Jana; Cizkova, Dasa

    2018-04-01

    Objectives In this study, a new approach was used with an in vitro model in which neural cells were exposed to conditioned media from the injured spinal cord (SCI-CM) mimicking a local inflammatory microenvironment . Subsequently, the neuroprotective effect of rat adipose tissue-derived msesenchymal stem cell-conditioned media (ATMSC-CM) was investigated through a cell-free based therapy, which was used to treat cortical neurons and astrocytes under inflammation. Methods Primary cell cultures isolated from postnatal day (P6) Wistar rat brain cortex were exposed to SCI-CM derived from the central lesion, rostral and caudal segments of injured spinal cord. After 48 h incubation, the SCI-CM was replaced and primary cultures were cultivated either in DMEM media alone or in ATMSC-CM for 72 h. The impact of ATMSC-CM on the viability of neurons and astrocytes was assessed using a CyQUANT® Direct Cell Proliferation Assay Kit as well as immunocytochemistry analysis. Results Immunocytochemical analysis revealed significant decrease in the number of MAP2 positive neurons exposed to SCI-CM compared to Control. Protection by ATMSC-CM was associated with increased survival of neurons compared to primary culture cultivated in DMEM media alone. The ATMSC-CM effect on astrocytes was more variable and without any significant impact. Conclusion The results demonstrate that SCI-CM mimicking inflammation can reduce cortical neuron survival, and subsequent exposure to ATMSC-CM can stabilize the neuronal population most likely via released neuroprotective and trophic factors. In addition, astrogliosis was not affected by ATMSC-CM.

  15. Vaginal Lactobacillus Inhibits HIV-1 Replication in Human Tissues Ex Vivo

    PubMed Central

    Ñahui Palomino, Rogers A.; Zicari, Sonia; Vanpouille, Christophe; Vitali, Beatrice; Margolis, Leonid

    2017-01-01

    Lactobacillus species, which dominate vaginal microbiota of healthy reproductive-age women, lower the risks of sexually transmitted infections, including the risk of human immunodeficiency virus (HIV) acquisition. The exact mechanisms of this protection remain to be understood. Here, we investigated these mechanisms in the context of human cervico-vaginal and lymphoid tissues ex vivo. We found that all six Lactobacillus strains tested in these systems significantly suppressed HIV type-1 (HIV-1) infection. We identified at least three factors that mediated this suppression: (i) Acidification of the medium. The pH of the undiluted medium conditioned by lactobacilli was between 3.8 and 4.6. Acidification of the culture medium with hydrochloric acid (HCl) to this pH in control experiments was sufficient to abrogate HIV-1 replication. However, the pH of the Lactobacillus-conditioned medium (CM) diluted fivefold, which reached ∼6.9, was also suppressive for HIV-1 infection, while in control experiments HIV-1 infection was not abrogated when the pH of the medium was brought to 6.9 through the use of HCl. This suggested the existence of other factors responsible for HIV-1 inhibition by lactobacilli. (ii) Lactic acid. There was a correlation between the concentration of lactic acid in the Lactobacillus-CM and its ability to suppress HIV-1 infection in human tissues ex vivo. Addition of lactic acid isomers D and L to tissue culture medium at the concentration that corresponded to their amount released by lactobacilli resulted in HIV-1 inhibition. Isomer L was produced in higher quantities than isomer D and was mostly responsible for HIV-1 inhibition. These results indicate that lactic acid, in particular its L-isomer, inhibits HIV-1 independently of lowering of the pH. (iii) Virucidal effect. Incubation of HIV-1 in Lactobacillus-CM significantly suppressed viral infectivity for human tissues ex vivo. Finally, lactobacilli adsorb HIV-1, serving as a sink decreasing the number of free virions. In summary, we found that lactobacilli inhibit HIV-1 replication in human tissue ex vivo by multiple mechanisms. Further studies are needed to evaluate the potential of altering the spectra of vaginal microbiota as an effective strategy to enhance vaginal health. Human tissues ex vivo may serve as a test system for these strategies. PMID:28579980

  16. Two Bioactive Molecular Weight Fractions of a Conditioned Medium Enhance RPE Cell Survival on Age-Related Macular Degeneration and Aged Bruch's Membrane

    PubMed Central

    Sugino, Ilene K.; Sun, Qian; Springer, Carola; Cheewatrakoolpong, Noounanong; Liu, Tong; Li, Hong; Zarbin, Marco A.

    2016-01-01

    Purpose To characterize molecular weight fractions of bovine corneal endothelial cell conditioned medium (CM) supporting retinal pigment epithelium (RPE) cell survival on aged and age-related macular degeneration (AMD) Bruch's membrane. Methods CM was subject to size separation using centrifugal filters. Retentate and filtrate fractions were tested for bioactivity by analyzing RPE survival on submacular Bruch's membrane of aged and AMD donor eyes and behavior on collagen I-coated tissue culture wells. Protein and peptide composition of active fractions was determined by mass spectrometry. Results Two bioactive fractions, 3-kDa filtrate and a 10-50–kDa fraction, were necessary for RPE survival on aged and AMD Bruch's membrane. The 3-kDa filtrate, but not the 10-50–kDa fraction, supported RPE growth on collagen 1‐coated tissue culture plates. Mass spectrometry of the 10-50–kDa fraction identified 175 extracellular proteins, including growth factors and extracellular matrix molecules. Transforming growth factor (TGF)β-2 was identified as unique to active CM. Peptides representing 29 unique proteins were identified in the 3-KDa filtrate. Conclusions These results indicate there is a minimum of two bioactive molecules in CM, one found in the 3-kDa filtrate and one in the 10-50–kDa fraction, and that bioactive molecules in both fractions must be present to ensure RPE survival on Bruch's membrane. Mass spectrometry analysis suggested proteins to test in future studies to identify proteins that may contribute to CM bioactivity. Translational Relevance Results of this study are the first steps in development of an adjunct to cell-based therapy to ensure cell transplant survival and functionality in AMD patients. PMID:26933521

  17. Human neuroblastoma growth inhibitory factor (h-NGIF), derived from human astrocytoma conditioned medium, has neurotrophic properties.

    PubMed

    Eksioglu, Y Z; Iida, J; Asai, K; Ueki, T; Nakanishi, K; Isobe, I; Yamagata, K; Kato, T

    1994-05-02

    Investigations on the general characteristics of human astrocytoma cell line NAC-1 revealed neuroblastoma growth inhibitory activity in conditioned medium. Neuroblastoma growth inhibitory factor (NGIF) was partially purified by Econo Q, Econo CM, and Superose 12 column chromatography. The protein is weakly basic with an estimated M(r) of 120,000, possibly having an M(r) 60,000 dimeric structure. NGIF inhibits the growth of human neuroblastoma cell lines but has no effect on morphology nor does it produce any change in the growth of human glioblastoma cell lines. Interestingly, NGIF appears to promote survival and neurite outgrowth of embryonal rat cortical neurons. These neurotrophic properties suggest a role for NGIF in the development of the nervous system.

  18. Differentiation of Mesenchymal Stem Cells from Human Umbilical Cord Tissue into Odontoblast-Like Cells Using the Conditioned Medium of Tooth Germ Cells In Vitro

    PubMed Central

    Li, Tian Xia; Yuan, Jie; Chen, Yan; Pan, Li Jie; Song, Chun; Bi, Liang Jia; Jiao, Xiao Hui

    2013-01-01

    The easily accessible mesenchymal stem cells in the Wharton's jelly of human umbilical cord tissue (hUCMSCs) have excellent proliferation and differentiation potential, but it remains unclear whether hUCMSCs can differentiate into odontoblasts. In this study, mesenchymal stem cells were isolated from the Wharton's jelly of human umbilical cord tissue using the simple method of tissue blocks culture attachment. UCMSC surface marker expression was then evaluated for the isolated cells using flow cytometry. The third-passage hUCMSCs induced by conditioned medium from developing tooth germ cells (TGC-CM) displayed high alkaline phosphatase (ALP) levels (P < 0.001), an enhanced ability to proliferate (P < 0.05), and the presence of mineralized nodules. These effects were not observed in cells treated with regular medium. After induction of hUCMSCs, the results of reverse transcriptional polymerase chain reaction (PCR) indicated that the dentin sialophosphoprotein (DSPP) and dentin matrix protein 1 (DMP1) genes were significantly tested. Additionally, dentin sialoprotein (DSP) and DMP1 demonstrated significant levels of staining in an immunofluorescence analysis. In contrast, the control cells failed to display the characteristics of odontoblasts. Taken together, these results suggest that hUCMSCs can be induced to differentiate into odontoblast-like cells with TGC-CM and provide a novel strategy for tooth regeneration research. PMID:23762828

  19. Oxalobacter formigenes–Derived Bioactive Factors Stimulate Oxalate Transport by Intestinal Epithelial Cells

    PubMed Central

    Arvans, Donna; Jung, Yong-Chul; Antonopoulos, Dionysios; Koval, Jason; Granja, Ignacio; Bashir, Mohamed; Karrar, Eltayeb; Roy-Chowdhury, Jayanta; Musch, Mark; Asplin, John; Chang, Eugene

    2017-01-01

    Hyperoxaluria is a major risk factor for kidney stones and has no specific therapy, although Oxalobacter formigenes colonization is associated with reduced stone risk. O. formigenes interacts with colonic epithelium and induces colonic oxalate secretion, thereby reducing urinary oxalate excretion, via an unknown secretagogue. The difficulties in sustaining O. formigenes colonization underscore the need to identify the derived factors inducing colonic oxalate secretion. We therefore evaluated the effects of O. formigenes culture conditioned medium (CM) on apical 14C-oxalate uptake by human intestinal Caco-2-BBE cells. Compared with control medium, O. formigenes CM significantly stimulated oxalate uptake (>2.4-fold), whereas CM from Lactobacillus acidophilus did not. Treating the O. formigenes CM with heat or pepsin completely abolished this bioactivity, and selective ultrafiltration of the CM revealed that the O. formigenes–derived factors have molecular masses of 10–30 kDa. Treatment with the protein kinase A inhibitor H89 or the anion exchange inhibitor 4,4'-diisothiocyano-2,2'-stilbenedisulfonic acid completely blocked the CM-induced oxalate transport. Knockdown of the oxalate transporter SLC26A6 also significantly restricted the induction of oxalate transport by CM. In a mouse model of primary hyperoxaluria type 1, rectal administration of O. formigenes CM significantly reduced (>32.5%) urinary oxalate excretion and stimulated (>42%) distal colonic oxalate secretion. We conclude that O. formigenes–derived bioactive factors stimulate oxalate transport in intestinal cells through mechanisms including PKA activation. The reduction in urinary oxalate excretion in hyperoxaluric mice treated with O. formigenes CM reflects the in vivo retention of biologic activity and the therapeutic potential of these factors. PMID:27738124

  20. Macrophage conditioned medium induced cellular network formation in MCF-7 cells through enhanced tunneling nanotube formation and tunneling nanotube mediated release of viable cytoplasmic fragments

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

    Patheja, Pooja, E-mail: pooja.patheja8@gmail.com; Homi Bhabha National Institute, Training School Complex, Anushaktinagar, Mumbai 400094, Maharashtra; Sahu, Khageswar

    Infiltrating macrophages in tumor microenvironment, through their secreted cytokines and growth factors, regulate several processes of cancer progression such as cancer cell survival, proliferation, invasion, metastasis and angiogenesis. Recently, intercellular cytoplasmic bridges between cancer cells referred as tunneling nanotubes (TNTs) have been recognized as novel mode of intercellular communication between cancer cells. In this study, we investigated the effect of inflammatory mediators present in conditioned medium derived from macrophages on the formation of TNTs in breast adenocarcinoma cells MCF-7. Results show that treatment with macrophage conditioned medium (MφCM) not only enhanced TNT formation between cells but also stimulated the releasemore » of independently migrating viable cytoplasmic fragments, referred to as microplasts, from MCF-7 cells. Time lapse microscopy revealed that microplasts were released from parent cancer cells in extracellular space through formation of TNT-like structures. Mitochondria, vesicles and cytoplasm could be transferred from parent cell body to microplasts through connecting TNTs. The microplasts could also be resorbed into the parent cell body by retraction of the connecting TNTs. Microplast formation inhibited in presence cell migration inhibitor, cytochalasin-B. Notably by utilizing migratory machinery within microplasts, distantly located MCF-7 cells formed several TNT based intercellular connections, leading to formation of physically connected network of cells. Together, these results demonstrate novel role of TNTs in microplast formation, novel modes of TNT formation mediated by microplasts and stimulatory effect of MφCM on cellular network formation in MCF-7 cells mediated through enhanced TNT and microplast formation.« less

  1. Polyacrylamide medium for the electrophoretic separation of biomolecules

    DOEpatents

    Madabhushi, Ramakrishna S.; Gammon, Stuart A.

    2003-11-11

    A polyacryalmide medium for the electrophoretic separation of biomolecules. The polyacryalmide medium comprises high molecular weight polyacrylamides (PAAm) having a viscosity average molecular weight (M.sub.v) of about 675-725 kDa were synthesized by conventional red-ox polymerization technique. Using this separation medium, capillary electrophoresis of BigDye DNA sequencing standard was performed. A single base resolution of .about.725 bases was achieved in .about.60 minute in a non-covalently coated capillary of 50 .mu.m i.d., 40 cm effective length, and a filed of 160 V/cm at 40.degree. C. The resolution achieved with this formulation to separate DNA under identical conditions is much superior (725 bases vs. 625 bases) and faster (60 min. vs. 75 min.) to the commercially available PAAm, such as supplied by Amersham. The formulation method employed here to synthesize PAAm is straight-forward, simple and does not require cumbersome methods such as emulsion polymerizaiton in order to achieve very high molecular weights. Also, the formulation here does not require separation of PAAm from the reaction mixture prior to reconstituting the polymer to a final concentration. Furthermore, the formulation here is prepared from a single average mol. wt. PAAm as opposed to the mixture of two different average mo. wt. PAAm previously required to achieve high resolution.

  2. Protection of individual ash trees from emerald ash borer (Coleoptera: Buprestidae) with basal soil applications of imidacloprid.

    PubMed

    Smitley, D R; Rebek, E J; Royalty, R N; Davis, T W; Newhouse, K F

    2010-02-01

    We conducted field trials at five different locations over a period of 6 yr to investigate the efficacy of imidacloprid applied each spring as a basal soil drench for protection against emerald ash borer, Agrilus planipennis Fairmaire (Coleoptera: Buprestidae). Canopy thinning and emerald ash borer larval density were used to evaluate efficacy for 3-4 yr at each location while treatments continued. Test sites included small urban trees (5-15 cm diameter at breast height [dbh]), medium to large (15-65 cm dbh) trees at golf courses, and medium to large street trees. Annual basal drenches with imidacloprid gave complete protection of small ash trees for three years. At three sites where the size of trees ranged from 23 to 37 cm dbh, we successfully protected all ash trees beginning the test with <60% canopy thinning. Regression analysis of data from two sites reveals that tree size explains 46% of the variation in efficacy of imidacloprid drenches. The smallest trees (<30 cm dbh) remained in excellent condition for 3 yr, whereas most of the largest trees (>38 cm dbh) declined to a weakened state and undesirable appearance. The five-fold increase in trunk and branch surface area of ash trees as the tree dbh doubles may account for reduced efficacy on larger trees, and suggests a need to increase treatment rates for larger trees.

  3. The Alcohol Dehydrogenase Gene Family in Melon (Cucumis melo L.): Bioinformatic Analysis and Expression Patterns

    PubMed Central

    Jin, Yazhong; Zhang, Chong; Liu, Wei; Tang, Yufan; Qi, Hongyan; Chen, Hao; Cao, Songxiao

    2016-01-01

    Alcohol dehydrogenases (ADH), encoded by multigene family in plants, play a critical role in plant growth, development, adaptation, fruit ripening and aroma production. Thirteen ADH genes were identified in melon genome, including 12 ADHs and one formaldehyde dehydrogenease (FDH), designated CmADH1-12 and CmFDH1, in which CmADH1 and CmADH2 have been isolated in Cantaloupe. ADH genes shared a lower identity with each other at the protein level and had different intron-exon structure at nucleotide level. No typical signal peptides were found in all CmADHs, and CmADH proteins might locate in the cytoplasm. The phylogenetic tree revealed that 13 ADH genes were divided into three groups respectively, namely long-, medium-, and short-chain ADH subfamily, and CmADH1,3-11, which belongs to the medium-chain ADH subfamily, fell into six medium-chain ADH subgroups. CmADH12 may belong to the long-chain ADH subfamily, while CmFDH1 may be a Class III ADH and serve as an ancestral ADH in melon. Expression profiling revealed that CmADH1, CmADH2, CmADH10 and CmFDH1 were moderately or strongly expressed in different vegetative tissues and fruit at medium and late developmental stages, while CmADH8 and CmADH12 were highly expressed in fruit after 20 days. CmADH3 showed preferential expression in young tissues. CmADH4 only had slight expression in root. Promoter analysis revealed several motifs of CmADH genes involved in the gene expression modulated by various hormones, and the response pattern of CmADH genes to ABA, IAA and ethylene were different. These CmADHs were divided into ethylene-sensitive and –insensitive groups, and the functions of CmADHs were discussed. PMID:27242871

  4. Neural stem cells inhibit melanin production by activation of Wnt inhibitors.

    PubMed

    Hwang, Insik; Park, Ju-Hwang; Park, Hang-Soo; Choi, Kyung-Ah; Seol, Ki-Cheon; Oh, Seung-Ick; Kang, Seongman; Hong, Sunghoi

    2013-12-01

    Melanin for skin pigmentation is synthesized from tyrosine via an enzymatic cascade that is controlled by tyrosinase (TYR), tyrosinase-related protein 1 (TRP1), and dopachrome tautomerase/tyrosinase related protein 2 (Dct/TRP2), which are the targets of microphthalmia-associated transcription factor (MITF). MITF is a master regulator of pigmentation and a target of β-catenin in Wnt/β-catenin signaling during melanocyte differentiation. Stem cells have been used in skin pigmentation studies, but the mechanisms were not determined for the conditioned medium (CM)-mediated effects. In this study, the inhibition and mechanisms of melanin synthesis were elucidated in B16 melanoma cells and UV-B irradiated C57/BL-6 mice that were treated with human neural stem cell-conditioned medium (NSC-CM). B16-F10 melanoma cells (1.5×10(4)cells/well) and the shaved dorsal skin of mice were pretreated with various amount (5, 10, 20, 50, and 100%) of NSC-CM. Melanin contents and TYR activity were measured by a Spectramax spectrophotometer. The expression of TYR, TRP1, Dct/TRP2, MITF, β-catenin and Wnt inhibitors were evaluated by RT-PCR and western blot. The dorsal skin samples were analyzed by immunofluorescence with various antibodies and compared with that control of tissues. Marked decreases were evident in melanin content and TYR, TRP1, DCT/TRP2, MITF, and β-catenin expression in B16 cells and C57/BL-6 mice. NSC-CM negatively regulated Wnt/β-catenin signaling by decreasing the expression of β-catenin protein, which resulted from robust expression of Wnt inhibitors Dickkopf-1 (DKK1) and secreted frizzled-related protein 2 (sFRP2). These results demonstrate that NSC-CM suppresses melanin production in vitro and in vivo, suggesting that factors in NSC-CM may play an important role in deregulation of epidermal melanogenesis. Copyright © 2013 Japanese Society for Investigative Dermatology. All rights reserved.

  5. Light intensity and N/P nutrient affect the accumulation of lipid and unsaturated fatty acids by Chlorella sp.

    PubMed

    Guo, Xiaoyi; Su, Gaomin; Li, Zheng; Chang, Jingyu; Zeng, Xianhai; Sun, Yong; Lu, Yinghua; Lin, Lu

    2015-09-01

    In this study, different light intensities (80, 160, 240 and 320 μmol/m(2) s) and various mediums including control medium (CM), N/P rich medium (NPM), N rich medium (NM), and P rich medium (PM) were applied for cultivation of Chlorella sp. It was revealed that cultivation of Chlorella sp. in CM under the light intensity of 320 μmol/m(2) s led to a lipid content up to 30% enhancement, which was higher than the results of other cases. A rather high unsaturated fatty acid (UFA) content of 7.5% and unsaturated fatty acid/total fatty acid (UFA/TFA) ratio of 0.73 were obtained under 320 μmol/m(2) s in CM, indicating that the CM-320 system was applicable for the generation of UFA. Moreover, Chlorella sp. cultivated in PM under 320 μmol/m(2) s provided higher TFA content (7.3%), which was appropriate for biofuel production. Copyright © 2015 Elsevier Ltd. All rights reserved.

  6. The Effect of Conditioned Media of Adipose-Derived Stem Cells on Wound Healing after Ablative Fractional Carbon Dioxide Laser Resurfacing

    PubMed Central

    Zhou, Bing-Rong; Xu, Yang; Guo, Shi-Lei; Xu, Yan; Wang, Ying; Zhu, Fen; Wu, Di; Yin, Zhi-Qiang; Luo, Dan

    2013-01-01

    Objective. To evaluate the benefits of conditioned medium of Adipose-derived stem cells (ADSC-CM) on wound healing after fractional carbon dioxide laser resurfacing (FxCR) on human skin. Materials and Methods. Nineteen subjects were treated with FxCR on the bilateral inner arms. ADSC-CM was applied on FxCR site of one randomly selected arm. Transepidermal water loss (TEWL), skin color, and gross-elasticity of FxCR site on both arms were measured. Skin samples were taken by biopsy from three subjects 3 weeks after treatment for histopathological manifestations and mRNA expressions of procollagen types I and III, elastin genes were noted. Results. The index of erythema, melanin, and TEWL of the ADSC-CM-treated skin were significantly lower than those of the control side. The mRNA expression of type III procollagen in ADSC-CM-treated group at 3 weeks posttreatment was 2.6 times of that of the control group. Conclusion. Application of allograft ADSC-CM is an effective method for enhancing wound healing after FxCR, by reducing transient adverse effects such as erythema, hyperpigmentation, and increased TEWL. PMID:24381938

  7. Conditioned medium from bone marrow-derived mesenchymal stem cells inhibits vascular calcification through blockade of the BMP2-Smad1/5/8 signaling pathway.

    PubMed

    Wang, Shuangshuang; Hu, Siwang; Wang, Jian; Liu, Yahui; Zhao, Ruochi; Tong, Maoqing; Cui, Hanbin; Wu, Nan; Chen, Xiaomin

    2018-06-13

    Arterial calcification is associated with cardiovascular disease as a complication of advanced atherosclerosis and is a significant contributor to cardiovascular morbidity and mortality. Osteoblastic differentiation of vascular smooth muscle cells (VSMCs) plays an important role in arterial calcification and is characterized by cellular necrosis, inflammation, and lipoprotein and phospholipid complexes, especially in atherosclerotic calcification. The conditioned medium from bone marrow-derived mesenchymal stem cells (MSC-CM) is well known as a rich source of autologous cytokines and is universally used for tissue regeneration in current clinical medicine. Here, we demonstrate that MSC-CM inhibits beta-glycerophosphate (β-GP)-induced vascular calcification through blockade of the bone morphogenetic protein-2 (BMP2)-Smad1/5/8 signaling pathway. VSMC calcification was induced by β-GP followed by treatment with MSC-CM. Mineral deposition was assessed by Alizarin Red S staining. Intracellular calcium content was determined colorimetrically by the o-cresolphthalein complexone method and alkaline phosphatase (ALP) activity was measured by the para-nitrophenyl phosphate method. Expression of BMP2, BMPR1A, BMPR1B, BMPR2, msh homeobox 2 (Msx2), Runt-related transcription factor 2 (Runx2), and osteocalcin (OC), representative osteoblastic markers, was assessed using real-time polymerase chain reaction analysis while the protein expression of BMP2, Runx2, and phosphorylated Smad1/5/8 was detected by western blot analysis. Our data demonstrated that MSC-CM inhibits osteoblastic differentiation and mineralization of VSMCs as evidenced by decreased calcium content, ALP activity, and decreased expression of BMP-2, Runx2, Msx2, and OC. MSC-CM suppressed the expression of phosphorylated Smad1/5/8 and the β-GP-induced translocation from the cytoplasm to the nucleus. Further study demonstrated that human recombinant BMP-2 overcame the suppression of VSMC calcification by MSC-CM. MSC-CM may act as a novel therapy for VSMC calcification by mediating the BMP2-Smad1/5/8 signaling pathway.

  8. Macrophage conditioned medium induced cellular network formation in MCF-7 cells through enhanced tunneling nanotube formation and tunneling nanotube mediated release of viable cytoplasmic fragments.

    PubMed

    Patheja, Pooja; Sahu, Khageswar

    2017-06-15

    Infiltrating macrophages in tumor microenvironment, through their secreted cytokines and growth factors, regulate several processes of cancer progression such as cancer cell survival, proliferation, invasion, metastasis and angiogenesis. Recently, intercellular cytoplasmic bridges between cancer cells referred as tunneling nanotubes (TNTs) have been recognized as novel mode of intercellular communication between cancer cells. In this study, we investigated the effect of inflammatory mediators present in conditioned medium derived from macrophages on the formation of TNTs in breast adenocarcinoma cells MCF-7. Results show that treatment with macrophage conditioned medium (MɸCM) not only enhanced TNT formation between cells but also stimulated the release of independently migrating viable cytoplasmic fragments, referred to as microplasts, from MCF-7 cells. Time lapse microscopy revealed that microplasts were released from parent cancer cells in extracellular space through formation of TNT-like structures. Mitochondria, vesicles and cytoplasm could be transferred from parent cell body to microplasts through connecting TNTs. The microplasts could also be resorbed into the parent cell body by retraction of the connecting TNTs. Microplast formation inhibited in presence cell migration inhibitor, cytochalasin-B. Notably by utilizing migratory machinery within microplasts, distantly located MCF-7 cells formed several TNT based intercellular connections, leading to formation of physically connected network of cells. Together, these results demonstrate novel role of TNTs in microplast formation, novel modes of TNT formation mediated by microplasts and stimulatory effect of MɸCM on cellular network formation in MCF-7 cells mediated through enhanced TNT and microplast formation. Copyright © 2017 Elsevier Inc. All rights reserved.

  9. Wnt/β-Catenin Pathway Regulates Cementogenic Differentiation of Adipose Tissue-Deprived Stem Cells in Dental Follicle Cell-Conditioned Medium

    PubMed Central

    Nie, Xin; Zhang, Bo; Zhou, Xia; Deng, Manjing

    2014-01-01

    The formation and attachment of new cementum is crucial for periodontium regeneration. Tissue engineering is currently explored to achieve complete, reliable and reproducible regeneration of the periodontium. The capacity of multipotency and self-renewal makes adipose tissue-deprived stem cells (ADSCs) an excellent cell source for tissue regeneration and repair. After rat ADSCs were cultured in dental follicle cell-conditioned medium (DFC-CM) supplemented with DKK-1, an inhibitor of the Wnt pathway, followed by 7 days of induction, they exhibited several phenotypic characteristics of cementoblast lineages, as indicated by upregulated expression levels of CAP, ALP, BSP and OPN mRNA, and accelerated expression of BSP and CAP proteins. The Wnt/β-catenin signaling pathway controls differentiation of stem cells by regulating the expression of target genes. Cementoblasts share phenotypical features with osteoblasts. In this study, we demonstrated that culturing ADSCs in DFC-CM supplemented with DKK-1 results in inhibition of β-catenin nuclear translocation and down-regulates TCF-4 and LEF-1 mRNA expression levels. We also found that DKK-1 could promote cementogenic differentiation of ADSCs, which was evident by the up-regulation of CAP, ALP, BSP and OPN gene expressions. On the other hand, culturing ADSCs in DFC-CM supplemented with 100 ng/mL Wnt3a, which activates the Wnt/β-catenin pathway, abrogated this effect. Taken together, our study indicates that the Wnt/β-catenin signaling pathway plays an important role in regulating cementogenic differentiation of ADSCs cultured in DFC-CM. These results raise the possibility of using ADSCs for periodontal regeneration by modifying the Wnt/β-catenin pathway. PMID:24806734

  10. Conditioned medium from LS 174T goblet cells treated with oxyresveratrol strengthens tight junctions in Caco-2 cells.

    PubMed

    Hwang, Dahyun; Jo, HyunA; Hwang, Seonwook; Kim, Jeong-Keun; Kim, In-Ho; Lim, Young-Hee

    2017-01-01

    Strengthening of intestinal tight junctions provides an effective barrier from the external environment. Goblet cell-derived trefoil factor 3 (TFF3) increases transepithelial resistance by upregulating the expression of tight junction proteins. Oxyresveratrol (OXY) is a hydroxyl-substituted stilbene found in the roots, leaves, stems, and fruit of many plants and known to have various biological activities. In this study, we investigated the strengthening effect of OXY on intestinal tight junctions through stimulation of TFF production in goblet cells. We prepared conditioned medium from LS 174T goblet cells treated with OXY (GCO-CM) and investigated the effect of GCO-CM on strengthening tight junctions of Caco-2 cells. The mRNA and protein expression levels of major tight junction components (claudin-1, occludin, and ZO-1) were measured by quantitative real-time PCR and western blotting, respectively. Transepithelial electric resistance (TEER) was measured using an ohm/V meter. Monolayer permeability was evaluated by paracellular transport of fluorescein isothiocyanate-dextran. OXY showed a strong antioxidant activity. It significantly increased the expression level of TFF3 in LS 174T goblet cells. GCO-CM prepared by treatment with 2.5, 5, and 10μg/ml OXY did not show cytotoxicity in Caco-2 cells. GCO-CM increased the mRNA and protein expression levels of claudin-1, occludin, and ZO-1. It also significantly increased tight junction integrity and reduced permeability in a dose-dependent manner. OXY stimulates the expression of TFF3 in goblet cells, which might increase the integrity of the intestinal tight junction barrier. Copyright © 2016 Elsevier Masson SAS. All rights reserved.

  11. Processes of interaction of laser radiation with porous transparent materials during their ablation

    NASA Astrophysics Data System (ADS)

    Osipov, V. V.; Lisenkov, V. V.; Platonov, V. V.; Tikhonov, E. V.

    2018-03-01

    The effect of ytterbium fibre laser radiation with λ = 1.07 μm on the pressed micropowders of transparent oxides and fluorides (CaF2, Y2O3, Al2O3, YSZ, etc.) having low absorption coefficients is theoretically studied. Using the numerical modelling, it is established that the radiation scattering in the medium of particles 0.5–4.6 μm in diameter leads to the concentration of radiation in the local regions of the medium, with the intensity level exceeding the intensity of the incident radiation (0.46 MW cm-2) by many times. It is shown that with the growth of the refractive index n of the material from 1.38 (MgF2) to 2.12 (YSZ) the intensity of radiation in the region of the strongest local maximum increases from 3.8 to 31 MW cm-2. In our opinion, this factor can be dominant in determining the possibility of achieving the ablation of such a medium by means of the radiation having an intensity of no greater than 1 MW cm-2. This hypothesis is experimentally checked by the example of preparing a nanopowder of CaF2 (n = 1.43) and 1% Nd : Y2O3 (n = 1.91) by evaporating the pressed target with the porosity 50% by the continuous-wave radiation of the ytterbium fibre laser with the power 600 W and the intensity 0.4 MW cm-2. Inside the CaF2 target, the scattered radiation intensity is smaller than the optical breakdown threshold, and the target is not evaporated. Under the same conditions, the target made of higher-melting 1% Nd : Y2O3 evaporates well, and the efficiency of the nanopowder production amounts to 23 g h-1.

  12. Excitation of Molecular Hydrogen in the Orion Bar Photodissociation Region from a Deep Near-infrared IGRINS Spectrum

    NASA Astrophysics Data System (ADS)

    Kaplan, Kyle F.; Dinerstein, Harriet L.; Oh, Heeyoung; Mace, Gregory N.; Kim, Hwihyun; Sokal, Kimberly R.; Pavel, Michael D.; Lee, Sungho; Pak, Soojong; Park, Chan; Sok Oh, Jae; Jaffe, Daniel T.

    2017-04-01

    We present a deep near-infrared spectrum of the Orion Bar Photodissociation Region (PDR) taken with the Immersion Grating INfrared Spectrometer (IGRINS) on the 2.7 m telescope at the McDonald Observatory. IGRINS has high spectral resolution (R˜ {{45,000}}) and instantaneous broad wavelength coverage (1.45-2.45 μm), enabling us to detect 87 emission lines from rovibrationally excited molecular hydrogen (H2) that arise from transitions out of 69 upper rovibration levels of the electronic ground state. These levels cover a large range of rotational and vibrational quantum numbers and excitation energies, making them excellent probes of the excitation mechanisms of H2 and physical conditions within the PDR. The Orion Bar PDR is thought to consist of cooler high density clumps or filaments (T=50{--}250 K, {n}H={10}5{--}{10}7 cm-3) embedded in a warmer lower density medium (T=250{--}1000 K, {n}H={10}4{--}{10}5 cm-3). We fit a grid of constant temperature and density Cloudy models, which recreate the observed H2 level populations well, to constrain the temperature to a range of 600-650 K and the density to {n}H=2.5× {10}3{--}{10}4 cm-3. The best-fit model gives T = 625 K and {n}H=5× {10}3 cm-3. This well-constrained warm temperature is consistent with kinetic temperatures found by other studies for the Orion Bar’s lower density medium. However, the range of densities well fit by the model grid is marginally lower than those reported by other studies. We could be observing lower density gas than the surrounding medium, or perhaps a density-sensitive parameter in our models is not properly estimated.

  13. Mesenchymal stem cells protect podocytes from apoptosis induced by high glucose via secretion of epithelial growth factor

    PubMed Central

    2013-01-01

    Introduction The apoptosis and subsequent injury of podocytes plays a pathogenic role in diabetic nephropathy (DN). Mesenchymal stem cells (MSCs) are promising therapeutic cells for preventing apoptosis and reducing cellular injury. Our previous study found that MSCs could protect kidneys from diabetes-induced injury without obvious engraftment. So we evaluated the effects of human adipose-derived MSCs (hAd-MSCs) on podocytic apoptosis and injury induced by high glucose (HG) and the underlying mechanisms. Methods We used flow cytometry, Western blot and confocal fluorescence microscopy to study podocytic apoptosis and injury induced by HG at 24 hours, 48 hours, and 72 hours in the presence or absence of MSC-conditioned medium (CM). An antibody-based cytokine array was used to identify the mediating factor, which was verified by adding the neutralizing antibody (NtAb) to block its function or adding the recombinant cytokine to the medium to induce its function. Results hAd-MSC-CM reduced podocytic apoptosis in a dose-dependent manner, decreased the expression of podocytic cleaved caspase-3, and prevented the reduced expression and maintained the normal arrangement of podocytic synaptopodin and nephrin. However, human embryonic lung cell (Wi38)-CM failed to ameliorate podocytic apoptosis or injury. Twelve cytokines with concentration ratios (MSC-CM/Wi38-CM) >10-fold were identified. Epithelial growth factor (EGF) was singled out for its known ability to prevent apoptosis. Recombinant human EGF (rhEGF) prevented podocytic apoptosis and injury similarly to hAd-MSC-CM but, upon blockade of EGF, the beneficial effect of hAd-MSC-CM decreased dramatically. Conclusions hAd-MSCs prevent podocytic apoptosis and injury induced by HG, mainly through secreting soluble EG. PMID:24004644

  14. Mesenchymal stem cells protect podocytes from apoptosis induced by high glucose via secretion of epithelial growth factor.

    PubMed

    Li, Diangeng; Wang, Nan; Zhang, Li; Hanyu, Zhu; Xueyuan, Bai; Fu, Bo; Shaoyuan, Cui; Zhang, Weiguang; Xuefeng, Sun; Li, Rongshan; Chen, Xiangmei

    2013-01-01

    The apoptosis and subsequent injury of podocytes plays a pathogenic role in diabetic nephropathy (DN). Mesenchymal stem cells (MSCs) are promising therapeutic cells for preventing apoptosis and reducing cellular injury. Our previous study found that MSCs could protect kidneys from diabetes-induced injury without obvious engraftment. So we evaluated the effects of human adipose-derived MSCs (hAd-MSCs) on podocytic apoptosis and injury induced by high glucose (HG) and the underlying mechanisms. We used flow cytometry, Western blot and confocal fluorescence microscopy to study podocytic apoptosis and injury induced by HG at 24 hours, 48 hours, and 72 hours in the presence or absence of MSC-conditioned medium (CM). An antibody-based cytokine array was used to identify the mediating factor, which was verified by adding the neutralizing antibody (NtAb) to block its function or adding the recombinant cytokine to the medium to induce its function. hAd-MSC-CM reduced podocytic apoptosis in a dose-dependent manner, decreased the expression of podocytic cleaved caspase-3, and prevented the reduced expression and maintained the normal arrangement of podocytic synaptopodin and nephrin. However, human embryonic lung cell (Wi38)-CM failed to ameliorate podocytic apoptosis or injury. Twelve cytokines with concentration ratios (MSC-CM/Wi38-CM) >10-fold were identified. Epithelial growth factor (EGF) was singled out for its known ability to prevent apoptosis. Recombinant human EGF (rhEGF) prevented podocytic apoptosis and injury similarly to hAd-MSC-CM but, upon blockade of EGF, the beneficial effect of hAd-MSC-CM decreased dramatically. hAd-MSCs prevent podocytic apoptosis and injury induced by HG, mainly through secreting soluble EG.

  15. Mapping low-frequency carbon radio recombination lines towards Cassiopeia A at 340, 148, 54, and 43 MHz

    NASA Astrophysics Data System (ADS)

    Salas, P.; Oonk, J. B. R.; van Weeren, R. J.; Wolfire, M. G.; Emig, K. L.; Toribio, M. C.; Röttgering, H. J. A.; Tielens, A. G. G. M.

    2018-04-01

    Quantitative understanding of the interstellar medium requires knowledge of its physical conditions. Low-frequency carbon radio recombination lines (CRRLs) trace cold interstellar gas and can be used to determine its physical conditions (e.g. electron temperature and density). In this work, we present spatially resolved observations of the low-frequency (≤390 MHz) CRRLs centred around C268α, C357α, C494α, and C539α towards Cassiopeia A on scales of ≤1.2 pc. We compare the spatial distribution of CRRLs with other interstellar medium tracers. This comparison reveals a spatial offset between the peak of the CRRLs and other tracers, which is very characteristic for photodissociation regions and that we take as evidence for CRRLs being preferentially detected from the surfaces of molecular clouds. Using the CRRLs, we constrain the gas electron temperature and density. These constraints on the gas conditions suggest variations of less than a factor of 2 in pressure over ˜1 pc scales, and an average hydrogen density of 200-470 cm-3. From the electron temperature and density maps, we also constrain the ionized carbon emission measure, column density, and path length. Based on these, the hydrogen column density is larger than 1022 cm-2, with a peak of ˜4 × 1022 cm-2 towards the south of Cassiopeia A. Towards the southern peak, the line-of-sight length is ˜40 pc over a ˜2 pc wide structure, which implies that the gas is a thin surface layer on a large (molecular) cloud that is only partially intersected by Cassiopeia A. These observations highlight the utility of CRRLs as tracers of low-density extended H I and CO-dark gas halo's around molecular clouds.

  16. Surface Molecules Released by Trypanosoma cruzi Metacyclic Forms Downregulate Host Cell Invasion

    PubMed Central

    Clemente, Tatiana Mordente; Cortez, Cristian; Novaes, Antônio da Silva; Yoshida, Nobuko

    2016-01-01

    Background The question whether metacylic trypomastigote (MT) forms of different T. cruzi strains differentially release surface molecules, and how they affect host cell invasion, remains to be fully clarified. We addressed that question using T. cruzi strains that differ widely in the ability to invade cells. Methodology/Principal Findings Metacyclic forms were incubated at 37°C for 1 h in complete D10 medium or in nutrient-deprived PBS containing Ca2+ and Mg2+ (PBS++). The conditioned medium (CM), collected after parasite centrifugation, was used for cell invasion assays and Western blot analysis, using monoclonal antibodies directed to gp82 and gp90, the MT surface molecules that promote and negatively regulate invasion, respectively. CM of poorly invasive G strain (G-CM) contained high amounts of gp90 and gp82, either in vesicles or as soluble molecules. CM of highly invasive CL strain (CL-CM) contained gp90 and gp82 at very low levels. HeLa cells were incubated for 1 h with CL strain MT in D10, in absence or in the presence of G-CM or CL-CM. Parasite invasion was significantly inhibited by G-CM, but not by CL-CM. As G strain MT invasion rate in D10 is very low, assays with this strain were performed in PBS++, which induces invasion-promoting lysosome-spreading. G-CM, but not CL-CM, significantly inhibited G strain internalization, effect that was counteracted by preincubating G-CM with an anti-gp90 monoclonal antibody or anti-gp82 polyclonal antibody that do not recognize live MT. G strain CM generated in PBS++ contained much lower amounts of gp90 and gp82 as compared to CM produced in D10, and exhibited lower inhibitory effect on host cell invasion. Conclusion/Significance Our data suggest that the surface molecules spontaneously released by MT impair parasite-host cell interaction, gp82 presumably competing with the molecule expressed on MT surface for the host cell receptor, and gp90 further contributing to down modulate invasion. PMID:27483135

  17. Hypoxic Conditioned Medium From Human Adipose-Derived Stem Cells Promotes Mouse Liver Regeneration Through JAK/STAT3 Signaling

    PubMed Central

    Lee, Sang Chul; Jeong, Hye Jin; Lee, Sang Kuon

    2016-01-01

    Adipose-derived stem cells (ASCs) mainly exert their function by secreting materials that are collectively termed the secretome. Despite recent attention to the secretome as an alternative to stem cell therapy, the culture conditions for generating optimal secretome contents have not been determined. Therefore, we investigated the role of hypoxic-conditioned media (HCM) from ASCs. Normoxic-conditioned media (NCM) and HCM were obtained after culturing ASCs in 20% O2 or 1% O2 for 24 hours, respectively. Subsequently, partially hepatectomized mice were infused with saline, control medium, NCM, or HCM, and then sera and liver specimens were obtained for analyses. Hypoxia (1% O2) significantly increased mRNA expression of mediators from ASCs, including interleukin-6 (IL-6), tumor necrosis factor α (TNF-α), hepatocyte growth factor (HGF), and vascular endothelial growth factor (VEGF). HCM infusion significantly increased the number of Ki67-positive cells in the liver (p < .05). HCM infusion significantly increased phospho-signal transducer and activator of transcription 3 (STAT3) and decreased suppressor of cytokine signaling 3 (SOCS3) expression in the liver (p < .05). To determine the role of IL-6 in liver regeneration, we then performed IL-6 RNA interference study. Conditioned media (CM) obtained from ASCs, which were transfected with either siIL-6 or siControl, were administered to partially hepatectomized mice. The siIL-6 CM groups exhibited lower liver proliferation (Ki67-positive cells) and markers of regeneration (protein expression of proliferating cell nuclear antigen, p-STAT3, HGF, and VEGF and liver weights) than the siControl CM groups (p < .05). Taken together, hypoxic preconditioning of ASCs increased expression of mediators promoting anti-inflammatory and regenerative responses. The liver regenerative effects of HCM appear to be mediated by persistent and uninhibited expression of STAT3 in the liver, which results from decreased expression of SOCS3. Significance In this study, it was found that treatment with the medium from hypoxic-preconditioned adipose-derived stem cells (ASCs) increased the viability of hepatotoxic hepatocytes and enhance liver regeneration in partially hepatectomized mice. In addition, the researchers first revealed that the hepatoprotective effects of hypoxic-conditioned media are mediated by persistent and uninhibited expression of signal transducer and activator of transcription 3 in the liver, which result from a decreased expression of suppressor of cytokine signaling 3. Therefore, the hypoxic preconditioning of ASCs is expected to play a crucial role in regenerative medicine by optimizing the production of a highly effective secretome from ASCs. PMID:27102647

  18. Differences in cytocompatibility, dynamics of the oxide layers' formation, and nickel release between superelastic and thermo-activated nickel-titanium archwires.

    PubMed

    Čolić, Miodrag; Tomić, Sergej; Rudolf, Rebeka; Marković, Evgenija; Šćepan, Ivana

    2016-08-01

    Superelastic (SE) and thermo-activated (TA) nickel-titanium (NiTi) archwires are used in everyday orthodontic practice, based on their acceptable biocompatibility and well-defined shape memory properties. However, the differences in their surface microstructure and cytotoxicity have not been clearly defined, and the standard cytotoxicity tests are too robust to detect small differences in the cytotoxicity of these alloys, all of which can lead to unexpected adverse reactions in some patients. Therefore, we tested the hypothesis that the differences in manufacture and microstructure of commercially available SE and TA archwires may influence their biocompatibility. The archwires were studied as-received and after conditioning for 24 h or 35 days in a cell culture medium under static conditions. All of the tested archwires, including their conditioned medium (CM), were non-cytotoxic for L929 cells, but Rematitan SE (both as received and conditioned) induced the apoptosis of rat thymocytes in a direct contact. In contrast, TruFlex SE and Equire TA increased the proliferation of thymocytes. The cytotoxic effect of Rematitan SE correlated with the higher release of Ni ions in CM, higher concentration of surface Ni and an increased oxygen layer thickness after the conditioning. In conclusion, the apoptosis assay on rat thymocytes, in contrast to the less sensitive standard assay on L929 cells, revealed that Rematitan SE was less cytocompatible compared to other archwires and the effect was most probably associated with a higher exposition of the cells to Ni on the surface of the archwire, due to the formation of unstable oxide layer.

  19. Impact of operating conditions on performance of a novel gas double-dynamic solid-state fermentation bioreactor (GDSFB).

    PubMed

    Chen, Hongzhang; Li, Yanjun; Xu, Fujian

    2013-11-01

    A self-designed novel solid-state fermentation (SSF) bioreactor named "gas double-dynamic solid-state fermentation bioreactor (GDSFB)" showed great success in processes for the production of several valuable products. For the present study, a simple GDSFB (2 L in volume) was designed to investigate the impact of exhaust time on SSF performance. Both air pressure and vent aperture significantly influenced the exhaust time. The production of cellulase by Penicillium decumbens JUA10 was studied in this bioreactor. When the vent aperture was maintained at 0.2 cm, the highest FPA activity of 17.2 IU/g dry solid-state medium was obtained at an air pressure of 0.2 MPa (gauge pressure). When the air pressure was maintained at 0.2 MPa, a vent aperture of 0.3 cm gave the highest FPA activity of 18.0 IU/g dry solid-state medium. Further analysis revealed that the exhaust time was a crucial indicator of good performance in GDSFB.

  20. Human Uterine Cervical Stromal Stem Cells (hUCESCs): Why and How they Exert their Antitumor Activity

    PubMed Central

    SCHNEIDER, JOSÉ; EIRÓ, NOEMÍ; PÉREZ-FERNÁNDEZ, ROMÁN; MARTÍNEZ-ORDÓÑEZ, ANXO; VIZOSO, FRANCISCO

    2016-01-01

    Our research team has recently isolated and characterized a new stromal stem cell line (hUCESCs) obtained from cytological smears, as routinely performed for cervical cancer screening. We have, furthermore, described that both hUCESCs directly, as well as the secretome contained in the conditioned medium used for growing them (hUCESCs-CM) have potent antitumoral, anti-inflammatory, antibiotic, antimycotic and re-epitheliasation-enhancing properties. The scientific explanation our team proposes for these pleiotropic effects are directly related to the site of origin of hUCESCs, the human cervical transition zone, which has unique features that biologically justify the different actions of hUCESCs and hUCESCs-CM. We, herein, expose our working theory for the biological activity of hUCESCs and hUCESCs-CM. PMID:27566652

  1. Iron-Binding Protein Degradation by Cysteine Proteases of Naegleria fowleri.

    PubMed

    Martínez-Castillo, Moisés; Ramírez-Rico, Gerardo; Serrano-Luna, Jesús; Shibayama, Mineko

    2015-01-01

    Naegleria fowleri causes acute and fulminant primary amoebic meningoencephalitis. This microorganism invades its host by penetrating the olfactory mucosa and then traveling up the mesaxonal spaces and crossing the cribriform plate; finally, the trophozoites invade the olfactory bulbs. During its invasion, the protozoan obtains nutrients such as proteins, lipids, carbohydrates, and cationic ions (e.g., iron, calcium, and sodium) from the host. However, the mechanism by which these ions are obtained, particularly iron, is poorly understood. In the present study, we evaluated the ability of N. fowleri to degrade iron-binding proteins, including hololactoferrin, transferrin, ferritin, and hemoglobin. Zymography assays were performed for each substrate under physiological conditions (pH 7 at 37°C) employing conditioned medium (CM) and total crude extracts (TCEs) of N. fowleri. Different degradation patterns with CM were observed for hololactoferrin, transferrin, and hemoglobin; however, CM did not cause ferritin degradation. In contrast, the TCEs degraded only hololactoferrin and transferrin. Inhibition assays revealed that cysteine proteases were involved in this process. Based on these results, we suggest that CM and TCEs of N. fowleri degrade iron-binding proteins by employing cysteine proteases, which enables the parasite to obtain iron to survive while invading the central nervous system.

  2. Iron-Binding Protein Degradation by Cysteine Proteases of Naegleria fowleri

    PubMed Central

    Ramírez-Rico, Gerardo; Serrano-Luna, Jesús; Shibayama, Mineko

    2015-01-01

    Naegleria fowleri causes acute and fulminant primary amoebic meningoencephalitis. This microorganism invades its host by penetrating the olfactory mucosa and then traveling up the mesaxonal spaces and crossing the cribriform plate; finally, the trophozoites invade the olfactory bulbs. During its invasion, the protozoan obtains nutrients such as proteins, lipids, carbohydrates, and cationic ions (e.g., iron, calcium, and sodium) from the host. However, the mechanism by which these ions are obtained, particularly iron, is poorly understood. In the present study, we evaluated the ability of N. fowleri to degrade iron-binding proteins, including hololactoferrin, transferrin, ferritin, and hemoglobin. Zymography assays were performed for each substrate under physiological conditions (pH 7 at 37°C) employing conditioned medium (CM) and total crude extracts (TCEs) of N. fowleri. Different degradation patterns with CM were observed for hololactoferrin, transferrin, and hemoglobin; however, CM did not cause ferritin degradation. In contrast, the TCEs degraded only hololactoferrin and transferrin. Inhibition assays revealed that cysteine proteases were involved in this process. Based on these results, we suggest that CM and TCEs of N. fowleri degrade iron-binding proteins by employing cysteine proteases, which enables the parasite to obtain iron to survive while invading the central nervous system. PMID:26090408

  3. Human Wharton's jelly stem cell conditioned medium and cell-free lysate inhibit human osteosarcoma and mammary carcinoma cell growth in vitro and in xenograft mice.

    PubMed

    Gauthaman, Kalamegam; Fong, Chui-Yee; Arularasu, Suganya; Subramanian, Arjunan; Biswas, Arijit; Choolani, Mahesh; Bongso, Ariff

    2013-02-01

    Human Wharton's jelly stem cells (hWJSCs) were shown to inhibit the growth of human mammary carcinomas. It is not known whether cell-free secretions or lysates of hWJSCs do the same on different cancers. They may be less controversial than cells to regulatory bodies for clinical application. We examined the influence of hWJSC conditioned medium (hWJSC-CM) and cell-free lysate (hWJSC-CL) on two osteosarcoma cell lines (MG-63, SKES-1) in vitro and on human mammary carcinomas in immunodeficient mice. When exposed to hWJSC-CL, increased vacuolations in MG-63 and increased membrane fragmentation in SKES-1 cells were observed, with greater cell death in SKES-1. Exposure of SKES-1 and MG-63 cells to hWJSC-CL showed significant decreases in cell proliferation of 46.48 ± 6.66% and 24.32 ± 5.67% respectively compared to controls. MG-63 and SKES-1 cells were annexin V-FITC positive and SKES-1 TUNEL positive following treatment with hWJSC-CM and hWJSC-CL. MG-63 cells were positive and SKES-1 cells negative for anti-BECLIN-1 and anti-LC3B following treatment with hWJSC-CM and hWJSC-CL. RT-PCR showed that the pro-apoptotic BAX gene and the autophagy-related ATG-5 and BECLIN-1 genes were up-regulated while the anti-apoptotic BCL2 and SURVIVIN genes were down-regulated in MG-63 and SKES-1 cells treated with hWJSC-CM and hWJSC-CL. Injections of hWJSCs and hWJSC-CM into mammary carcinomas in immunodeficient mice resulted in decreased tumor sizes and weights of 24.86 ± 6.05% to 37.03 ± 5.91% and 47.14 ± 7.36% to 55.09 ± 5.87% respectively at 6 weeks compared to controls. hWJSC-CM and hWJSC-CL inhibit mammary carcinoma and osteosarcoma cells via apoptosis and autophagy. Copyright © 2012 Wiley Periodicals, Inc.

  4. NAA-Induced Direct Organogenesis from Female Immature Inflorescence Explants of Date Palm.

    PubMed

    Khierallah, Hussam S M; Bader, Saleh M; Al-Khafaji, Makki A

    2017-01-01

    Micropropagation has great potential for the multiplication of female and male date palms of commercially grown cultivars by using inflorescences. This approach is simple, convenient, and much faster than the conventional method of using shoot-tip explants. We describe here a stepwise micropropagation procedure using inflorescence explants of Iraqi date palm cultivar Maktoom. Cultured explants were derived from 0.5-cm-long spike segments excised from 8 to 10-cm-long spathes. About 70% formed adventitious buds on Murashige and Skoog (MS) medium supplemented with 2 mg/L naphthalene acetic acid (NAA), 4 mg/L benzylaminopurine (BAP), and 40 g/L sucrose and maintained in the dark for 16 weeks before transferring to normal light conditions. The best multiplication rate was achieved with 3 mg/L 2ip and 2 mg/L; for shoot elongation, the best medium is MS containing 0.5 mg/L BAP, 0.5 mg/L 2ip, and 1 mg/L GA 3 . Well-developed shoots were cultured for rooting in half MS medium amended with 1 mg/L NAA and 45 g/L sucrose. Plantlets with well-developed roots were successfully hardened in the greenhouse. Inflorescence explants proved to be a promising alternative explant source for micropropagation of date palm cultivars.

  5. Increased viability of odontoblast-like cells subjected to low-level laser irradiation

    NASA Astrophysics Data System (ADS)

    Oliveira, C. F.; Basso, F. G.; Lins, E. C.; Kurachi, C.; Hebling, J.; Bagnato, V. S.; de Souza Costa, C. A.

    2010-07-01

    Studies have shown that the increase of cell metabolism depends on the low level laser therapy (LLLT) parameters used to irradiate the cells. However, the optimal laser dose to up-regulate pulp cell activity remains unknown. Consequently, the aim of this study was to evaluate the metabolic response of odontoblast-like cells (MDPC-23) exposed to different LLLT doses. Cells at 20000 cells/cm2 were seeded in 24-well plates using plain culture medium (DMEM) and were incubated in a humidified incubator with 5% CO2 at 37°C. After 24 h, the culture medium was replaced by fresh DMEM supplemented with 5% (stress by nutritional deficit) or 10% fetal bovine serum (FBS). The cells were exposed to different laser doses from a near infrared diode laser prototype designed to provide a uniform irradiation of the wells. The experimental groups were: G1: 1.5 J/cm2 + 5% FBS; G2: 1.5 J/cm2 + 10% FBS; G3: 5 J/cm2 + 5% FBS; G4: 5 J/cm2 + 10% FBS; G5: 19 J/cm2 + 5% FBS; G6: 19 J/cm2 + 10% FBS. LLLT was performed in 3 consecutive irradiation cycles with a 24-hour interval. Non-irradiated cells cultured in DMEM supplemented with either 5 or 10% FBS served as control groups. The analysis of the metabolic response was performed by the MTT assay 3 h after the last irradiation. G1 presented an increase in SDH enzyme activity and differed significantly (Mann-Whitney test, p < 0.05) from the other groups. Analysis by scanning electron microscopy showed normal cell morphology in all groups. Under the tested conditions, LLLT stimulated the metabolic activity of MDPC-23 cultured in DMEM supplemented with 5% FBS and exposed to a laser dose of 1.5 J/cm2. These findings are relevant for further studies on the action of near infrared lasers on cells with odontoblast phenotype.

  6. SU-E-T-577: Commissioning of a Deterministic Algorithm for External Photon Beams

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

    Zhu, T; Finlay, J; Mesina, C

    Purpose: We report commissioning results for a deterministic algorithm for external photon beam treatment planning. A deterministic algorithm solves the radiation transport equations directly using a finite difference method, thus improve the accuracy of dose calculation, particularly under heterogeneous conditions with results similar to that of Monte Carlo (MC) simulation. Methods: Commissioning data for photon energies 6 – 15 MV includes the percentage depth dose (PDD) measured at SSD = 90 cm and output ratio in water (Spc), both normalized to 10 cm depth, for field sizes between 2 and 40 cm and depths between 0 and 40 cm. Off-axismore » ratio (OAR) for the same set of field sizes was used at 5 depths (dmax, 5, 10, 20, 30 cm). The final model was compared with the commissioning data as well as additional benchmark data. The benchmark data includes dose per MU determined for 17 points for SSD between 80 and 110 cm, depth between 5 and 20 cm, and lateral offset of up to 16.5 cm. Relative comparisons were made in a heterogeneous phantom made of cork and solid water. Results: Compared to the commissioning beam data, the agreement are generally better than 2% with large errors (up to 13%) observed in the buildup regions of the FDD and penumbra regions of the OAR profiles. The overall mean standard deviation is 0.04% when all data are taken into account. Compared to the benchmark data, the agreements are generally better than 2%. Relative comparison in heterogeneous phantom is in general better than 4%. Conclusion: A commercial deterministic algorithm was commissioned for megavoltage photon beams. In a homogeneous medium, the agreement between the algorithm and measurement at the benchmark points is generally better than 2%. The dose accuracy for a deterministic algorithm is better than a convolution algorithm in heterogeneous medium.« less

  7. An Index-Mismatch Scattering Approach to Optical Limiting

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

    Exarhos, Gregory J.; Ferris, Kim F.; Windisch, Charles F.

    A densely packed bed of alkaline earth fluoride particles percolated by a fluid medium has been investigated as a potential index-matched optical limiter in the spirit of a Christiansen-Shelyubskii filter. Marked optical limiting was observed through this transparent medium under conditions where the focused second-harmonic output of a Q-swtiched Nd: YAG laser was on the order of about 1 J/cm2. An open-aperture Z-scan technique was used to quantify the limiting behavior. In this case, the mechanism of optical limiting is thought to be a nonlinear shift in the fluid index of refraction, resulting in an index mismatch between the disparatemore » phases at high laser fluence.« less

  8. Impacts, recovery rates, and treatment options for spilled oil in marshes.

    PubMed

    Michel, Jacqueline; Rutherford, Nicolle

    2014-05-15

    In a review of the literature on impacts of spilled oil on marshes, 32 oil spills and field experiments were identified with sufficient data to generate recovery curves and identify influencing factors controlling the rate of recovery. For many spills, recovery occurred within 1-2 growing seasons, even in the absence of any treatment. Recovery was longest for spills with the following conditions: Cold climate; sheltered settings; thick oil on the marsh surface; light refined products with heavy loading; oils that formed persistent thick residues; and intensive treatment. Recovery was shortest for spills with the following conditions: Warm climate; light to heavy oiling of the vegetation only; medium crude oils; and less-intensive treatment. Recommendations are made for treatment based on the following oiling conditions: Free-floating oil on the water in the marsh; thicker oil (>0.5 cm) on marsh surface; thinner oil (<0.5 cm) on marsh surface; heavy oil loading on vegetation; and light to moderate oil loading on vegetation. Copyright © 2014 Elsevier Ltd. All rights reserved.

  9. Environmental impacts of different crop rotations in terms of soil compaction.

    PubMed

    Götze, Philipp; Rücknagel, Jan; Jacobs, Anna; Märländer, Bernward; Koch, Heinz-Josef; Christen, Olaf

    2016-10-01

    Avoiding soil compaction caused by agricultural management is a key aim of sustainable land management, and the soil compaction risk should be considered when assessing the environmental impacts of land use systems. Therefore this project compares different crop rotations in terms of soil structure and the soil compaction risk. It is based on a field trial in Germany, in which the crop rotations (i) silage maize (SM) monoculture, (ii) catch crop mustard (Mu)_sugar beet (SB)-winter wheat (WW)-WW, (iii) Mu_SM-WW-WW and (iv) SB-WW-Mu_SM are established since 2010. Based on the cultivation dates, the operation specific soil compaction risks and the soil compaction risk of the entire crop rotations are modelled at two soil depths (20 and 35 cm). To this end, based on assumptions of the equipment currently used in practice by a model farm, two scenarios are modelled (100 and 50% hopper load for SB and WW harvest). In addition, after one complete rotation, in 2013 and in 2014, the physical soil parameters saturated hydraulic conductivity (kS) and air capacity (AC) were determined at soil depths 2-8, 12-18, 22-28 and 32-38 cm in order to quantify the soil structure. At both soil depths, the modelled soil compaction risks for the crop rotations including SB (Mu_SB-WW-WW, SB-WW-Mu_SM) are higher (20 cm: medium to very high risks; 35 cm: no to medium risks) than for those without SB (SM monoculture, Mu_SM-WW-WW; 20 cm: medium risks; 35 cm: no to low risks). This increased soil compaction risk is largely influenced by the SB harvest in years where soil water content is high. Halving the hopper load and adjusting the tyre inflation pressure reduces the soil compaction risk for the crop rotation as a whole. Under these conditions, there are no to low soil compaction risks for all variants in the subsoil (soil depth 35 cm). Soil structure is mainly influenced in the topsoil (2-8 cm) related to the cultivation of Mu as a catch crop and WW as a preceding crop. Concerning kS, Mu_SB-WW-WW (240 cm d(-1)) and Mu_SM-WW-WW (196 cm d(-1)) displayed significantly higher values than the SM monoculture (67 cm d(-1)), indicating better structural stability and infiltration capacity. At other soil depths, and for the parameter AC, there are no systematic differences in soil structure between the variants. Under the circumstances described, all crop rotations investigated are not associated with environmental impacts caused by soil compaction. Copyright © 2016 Elsevier Ltd. All rights reserved.

  10. Hypoxia-inducible Factor-2α-dependent Hypoxic Induction of Wnt10b Expression in Adipogenic Cells*

    PubMed Central

    Park, Young-Kwon; Park, Bongju; Lee, Seongyeol; Choi, Kang; Moon, Yunwon; Park, Hyunsung

    2013-01-01

    Adipocyte hyperplasia and hypertrophy in obesity can lead to many changes in adipose tissue, such as hypoxia, metabolic dysregulation, and enhanced secretion of cytokines. In this study, hypoxia increased the expression of Wnt10b in both human and mouse adipogenic cells, but not in hypoxia-inducible factor (HIF)-2α-deficient adipogenic cells. Chromatin immunoprecipitation analysis revealed that HIF-2α, but not HIF-1α, bound to the Wnt10b enhancer region as well as upstream of the Wnt1 gene, which is encoded by an antisense strand of the Wnt10b gene. Hypoxia-conditioned medium (H-CM) induced phosphorylation of lipoprotein-receptor-related protein 6 as well as β-catenin-dependent gene expression in normoxic cells, which suggests that H-CM contains canonical Wnt signals. Furthermore, adipogenesis of both human mesenchymal stem cells and mouse preadipocytes was inhibited by H-CM even under normoxic conditions. These results suggest that O2 concentration gradients influence the formation of Wnt ligand gradients, which are involved in the regulation of pluripotency, cell proliferation, and cell differentiation. PMID:23900840

  11. Mosquito density, biting rate and cage size effects on repellent tests.

    PubMed

    Barnard, D R; Posey, K H; Smith, D; Schreck, C E

    1998-01-01

    Mosquito biting rates and the mean duration of protection (in hours) from bites (MDPB) of Aedes aegypti and Anopheles quadrimaculatus, using the repellent 'deet' (N,N-diethyl-3-methylbenzamide) on a 50 cm2 area of healthy human skin, were observed in small (27 l), medium (approximately 65 l) and large (125 l) cages containing low, medium or high densities of mosquitoes: respectively, 640, 128 or 49 cm3 of cage volume per female. At the initial treatment rate of approximately 0.4 microliter/cm2 (1 ml of 25% deet in ethanol on 650 cm2 of skin), the MDPB for deet against Ae. aegypti ranged from 4.5 to 6.5 h and was significantly less (5.0 +/- 0.8 h) in large cages compared with medium (6.2 +/- 0.9 h) and small (6.2 +/- 0.8 h) cages, regardless of the density. Against An. quadrimaculatus the MDPB for deet 0.4 microliter/cm2 was 1.5-8.0 h, less in small (3.7 +/- 2.3 h) and large (2.2 +/- 1.1 h) cages at medium (3.7 +/- 2.3 h) and high (2.5 +/- 1.7 h) mosquito densities, and was longest in medium cages (6.2 +/- 2.6 h) at low mosquito densities (5.8 +/- 2.8 h). With equinoxial photoperiodicity (light on 06.00-18.00 hours) the biting rate was influenced by the time of observation (08.00, 12.00, 16.00 hours) for Ae. aegypti but not for An. quadrimaculatus. For both species, the biting rate was inversely proportional to mosquito density and the MDPB. The shortest MDPBs were obtained in large cages with high densities of mosquitoes and longest protection times occurred in medium sized cages with low mosquito densities.

  12. Optical imaging of objects in turbid medium with ultrashort pulses

    NASA Astrophysics Data System (ADS)

    Wang, Chih-Yu; Sun, Chia-Wei; Yang, Chih Chung; Kiang, Yean-Woei; Lin, Chii-Wann

    2000-07-01

    Photons are seriously scattered when entering turbid medium; this the images of objects hidden in turbid medium can not be obtained by just collecting the transmitted photons. Early-arriving photons, which are also called ballistic or snake protons, are much less scattered when passing through turbid medium, and contains more image information than the late-arriving ones. Therefore, objects embedded in turbid medium can be imaged by gathering the ballistic and snake photons. In the present research we try to recover images of objects in turbid medium by simultaneously time-gate and polarization-gate to obtain the snake photons. An Argon-pumped Ti-Sapphire laser with 100fs pulses was employed as a light source. A streak camera with a 2ps temporal resolution was used to extract the ballistic and snake photons. Two pieces of lean swine meat, measured 4mmX3mm and 5xxX4mm, respectively, were placed in a 10cmX10cmX3cm acrylic tank, which was full of diluted milk. A pair of polarizer and an analyzer was used to extract the light that keeps polarization unchanged. The combination of time gating and polarization gating resulted in good images of objects hidden in turbid medium.

  13. Integration of Andrographis paniculata as Potential Medicinal Plant in Chir Pine (Pinus roxburghii Sarg.) Plantation of North-Western Himalaya

    PubMed Central

    Sanwal, Chandra Shekher; Bhardwaj, S. D.

    2016-01-01

    The integration of Andrographis paniculata under Pinus roxburghii (Chir pine) plantation has been studied to evaluate the growth and yield for its economic viability and conservation. It was grown on three topographical aspects, namely, northern, north-western, and western, at a spacing of 30 cm × 30 cm, followed by three tillage depths, namely, minimum (0 cm), medium (up to 10 cm), and deep (up to 15 cm) tillage. The growth parameters, namely, plant height and number of branches per plant, were recorded as significantly higher on western aspect and lowest on northern aspect except for leaf area index which was found nonsignificant. However under all tillage practices all the growth parameters in both understorey and open conditions were found to be nonsignificant except for plant height which was found to be significantly highest under deep tillage and lowest under minimum tillage. The study of net returns for Andrographis paniculata revealed that it had positive average annual returns even in understorey conditions which indicate its possible economic viability under integration of Chir pine plantations. Hence net returns can be enhanced by integrating Andrographis paniculata and this silvimedicinal system can be suggested which will help utilizing an unutilized part of land and increase total productivity from such lands besides conservation of the A. paniculata in situ. PMID:27563482

  14. Optimizing micropropagation of drought resistant Pyrus boissieriana Buhse.

    PubMed

    Zakavi, Maryam; Askari, Hossein; Irvani, Neda

    2016-10-01

    The present study concentrated on introducing a micropropagation protocol for a drought resistant genotype from Pyrus boissieriana , which is the second most naturally widespread pear species in Iran with proper physiological and medicinal properties. Proliferating microshoot cultures were obtained by placing nodal segments on MS medium supplemented with BAP and IBA or NAA. The highest number of shoots (27 shoots per explant) were obtained with 1.5 mg l -1 BAP and 0.05 mg l -1 IBA, but this combination did not produce shoots of desirable length (>1.7 cm). Combination of 1.75 mg l -1 BAP and 0.07 mg l -1 IBA was the best for the shoot multiplication in P. boissieriana with a sufficient number of shoot production (22.33 shoots per explant) and relatively more appropriate shoot length. The larger and greenish leaves were obtained when PG was added to the best multiplication treatment. Microshoot elongation was carried out in 1/2 and 1/4 MS medium containing 50-100 mg l -1 PG with different concentrations of IBA or NAA at intervals of 30-60 days. Significant increase in shoot length was detected after 45-60 days of culture in the presence of PG. The highest shoot length (8 cm) was recorded on 1/2 MS medium supplemented with 0.5 mg l -1 IBA and 100 mg l -1 PG. GA 3 negatively affected number and length of shoots and generally caused generation of red leaves. The highest percentage of root induction (100%) and root length (9 cm) were obtained on 1/6 strength MS medium supplemented with 0.005 mg l -1 IBA. All plantlets were hardened when transferred to ex vitro conditions through a period of 25-30 days. The results suggest axillary shoot proliferation of P. boissieriana could successfully be employed for propagation of candidate drought resistant seedling.

  15. Purification and culture of adult rat dorsal root ganglia neurons.

    PubMed

    Delree, P; Leprince, P; Schoenen, J; Moonen, G

    1989-06-01

    To study the trophic requirements of adult rat dorsal root ganglia neurons (DRG) in vitro, we developed a purification procedure that yields highly enriched neuronal cultures. Forty to fifty ganglia are dissected from the spinal column of an adult rat. After enzymatic and mechanical dissociation of the ganglia, myelin debris are eliminated by centrifugation on a Percoll gradient. The resulting cell suspension is layered onto a nylon mesh with a pore size of 10 microns. Most of the neurons, the diameter of which ranged from 17 microns to greater than 100 microns, are retained on the upper surface of the sieve; most of the non-neuronal cells with a caliber of less than 10 microns after trypsinization go through it. Recovery of neurons is achieved by reversing the mesh onto a Petri dish containing culture medium. Neurons to non-neurons ratio is 1 to 10 in the initial cell suspension and 1 to 1 after separation. When these purified neurons are seeded at a density of 3,000 neurons/cm2 in 6 mm polyornithine-laminin (PORN-LAM) coated wells, neuronal survival (assessed by the ability to extend neurites), measured after 48 hr of culture, is very low (from 0 to 16%). Addition of nerve growth factor (NGF) does not improve neuronal survival. However, when neurons are cultured in the presence of medium conditioned (CM) by astrocytes or Schwann cells, 60-80% of the seeded, dye-excluding neurons survive. So, purified adult DRG neurons require for their short-term survival and regeneration in culture, a trophic support that is present in conditioned medium from PNS or CNS glia.(ABSTRACT TRUNCATED AT 250 WORDS)

  16. Detection of the Galactic Warm Neutral Medium in HI 21cm absorption

    NASA Astrophysics Data System (ADS)

    Patra, Narendra Nath; Kanekar, Nissim; Chengalur, Jayaram N.; Roy, Nirupam

    2018-05-01

    We report a deep Giant Metrewave Radio Telescope (GMRT) search for Galactic HI 21cm absorption towards the quasar B0438-436, yielding the detection of wide, weak HI 21cm absorption, with a velocity-integrated HI 21cm optical depth of 0.0188 ± 0.0036 km s-1. Comparing this with the HI column density measured in the Parkes Galactic All-Sky Survey gives a column density-weighted harmonic mean spin temperature of 3760 ± 365 K, one of the highest measured in the Galaxy. This is consistent with most of the HI along the sightline arising in the stable warm neutral medium (WNM). The low peak HI 21cm optical depth towards B0438-436 implies negligible self-absorption, allowing a multi-Gaussian joint decomposition of the HI 21cm absorption and emission spectra. This yields a gas kinetic temperature of T_k ≤ (4910 ± 1900) K, and a spin temperature of T_s = (1000 ± 345) K for the gas that gives rise to the HI 21cm absorption. Our data are consistent with the HI 21cm absorption arising from either the stable WNM, with T_s ≪ T_k, T_k ≈ 5000 K, and little penetration of the background Lyman-α radiation field into the neutral hydrogen, or from the unstable neutral medium, with T_s ≈ T_k ≈ 1000K.

  17. Analysis of alkaline exchange membrane fuel cells performance at different operating conditions using DC and AC methods

    NASA Astrophysics Data System (ADS)

    Reshetenko, Tatyana; Odgaard, Madeleine; Schlueter, Debbie; Serov, Alexey

    2018-01-01

    Membrane electrode assemblies (MEAs) for anion exchange membrane fuel cells (AEMFCs) were manufactured from commercial materials: Pt/C catalyst, A201 AEM and AS4 ionomer by using an industrial mass-production digital printing method. The MEA designs selected are close to those recommended by US Department of Energy, including low loading of platinum on the cathode side (0.2 mg cm-2). Polarization curves and electrochemical impedance spectroscopy (EIS) were applied for MEA evaluation in fuel cell conditions with variation of gas humidification and oxygen partial pressure (air vs oxygen). The typical impedance curves recorded at H2/O2 gas configuration consist of high- and medium-frequency arcs responsible for hydrogen oxidation and oxygen reduction, respectively. Operation with air as a cathode feed gas resulted in a decrease in AEMFC performance due to possible CO2 poisoning and mass transfer losses. At the same time, EIS demonstrated formation of a low frequency loop due to diffusion limitations. Despite the low loading of platinum on the cathode (0.2 mg cm-2), a peak power density of ∼330 mW cm-2 was achieved (at 50/50% of RH on anode and cathode), which is substantially higher performance than for AEMFC MEAs tested at similar conditions.

  18. Human Uterine Cervical Stromal Stem Cells (hUCESCs): Why and How they Exert their Antitumor Activity.

    PubMed

    Schneider, José; Eiró, Noemí; Pérez-Fernández, Román; Martínez-Ordóñez, Anxo; Vizoso, Francisco

    Our research team has recently isolated and characterized a new stromal stem cell line (hUCESCs) obtained from cytological smears, as routinely performed for cervical cancer screening. We have, furthermore, described that both hUCESCs directly, as well as the secretome contained in the conditioned medium used for growing them (hUCESCs-CM) have potent antitumoral, anti-inflammatory, antibiotic, antimycotic and re-epitheliasation-enhancing properties. The scientific explanation our team proposes for these pleiotropic effects are directly related to the site of origin of hUCESCs, the human cervical transition zone, which has unique features that biologically justify the different actions of hUCESCs and hUCESCs-CM. We, herein, expose our working theory for the biological activity of hUCESCs and hUCESCs-CM. Copyright© 2016, International Institute of Anticancer Research (Dr. John G. Delinasios), All rights reserved.

  19. Hyperactive α-amylase production by Aspergillus oryzae IFO 30103 in a new bioreactor.

    PubMed

    Dey, T B; Banerjee, R

    2012-02-01

      To improve the α-amylase production in solid-state fermentation (SSF) condition utilizing a new bioreactor (NB) system.   In NB system, 20 g of wheat bran moistened with liquid medium in 1:1 ratio (w/v) was taken on the tray present inside the upper vessel and an additional 80 ml medium was supplemented into the lower vessel. Oxygen uptake rate was improved by supplying compressed air that lifted the liquid medium into the upper vessel and touched the substrate bed. This condition probably facilitated the heat transfer to liquid medium, reduce water loss and catabolite repression. With 1% glucose supplementation, maximum α-amylase activity of 22 317 Ugds(-1) was produced by Aspergillus oryzae IFO 30103 within a very short incubation period (48 h) at 2-cm bed height with air flow rate of 0·1 l min(-1)  g(-1) wheat bran at 32°C and initial medium pH of 6.   Within a short incubation period, significantly high α-amylase activity was obtained and it is higher than those reported to date at bioreactor scale operating with a fungal strain.   The reactor is novel and can overcome some of the major problems associated with SSF process. A. oryzae IFO 30103 is reported as the best fungal source for α-amylase production. © 2011 The Authors. Letters in Applied Microbiology © 2011 The Society for Applied Microbiology.

  20. Corrosive and cytotoxic properties of compact specimens and microparticles of Ni-Cr dental alloy.

    PubMed

    Ristic, Ljubisa; Vucevic, Dragana; Radovic, Ljubica; Djordjevic, Snezana; Nikacevic, Milutin; Colic, Miodrag

    2014-04-01

    Nickel-chromium (Ni-Cr) dental alloys have been widely used in prosthodontic practice, but there is a permanent concern about their biocompatibility due to the release of metal ions. This is especially important when Ni-Cr metal microparticles are incorporated into gingival tissue during prosthodontic procedures. Therefore, the aim of this study was to examine and compare the corrosion and cytotoxic properties of compact specimens and microparticles of Ni-Cr dental alloy. Ni-Cr alloy, Remanium CSe bars (4 mm diameter), were made by the standard casting method and then cut into 0.5-mm-thick disks. Metal particles were obtained by scraping the bars using a diamond instrument for crown preparation. The microstructure was observed by an optical microscope. Quantitative determination and morphological and dimensional characterization of metal particles were carried out by a scanning electron microscope and Leica Application Suite software for image analysis. Corrosion was studied by conditioning the alloy specimens in the RPMI 1640 medium, containing 10% fetal calf serum in an incubator with 5% CO2 for 72 hours at 37°C. Inductively coupled plasma-optical emission spectrometry was used to assess metal ion release. The cytotoxity of conditioning medium (CM) was investigated on L929 cells using an MTT test. One-way ANOVA was used for statistical analysis. After casting, the microstructure of the Remanium CSe compact specimen composed of Ni, Cr, Mo, Si, Fe, Al, and Co had a typical dendritic structure. Alloy microparticles had an irregular shape with a wide size range: from less than 1 μm to more than 100 μm. The release of metal ions, especially Ni and Mo from microparticles, was significantly higher, compared to the compact alloy specimen. The CM prepared from compact alloy was not cytotoxic at any tested dilutions, whereas CM from alloy microparticles showed dose-dependent cytotoxicity (90% CM and 45% CM versus control; p < 0.005). Ni-Cr microparticles showed less corrosion resistance and lower biocompatibility than compact alloy. This could affect health on long-term exposure, especially in sensitized individuals. © 2013 by the American College of Prosthodontists.

  1. Unveiling the Diffuse, Neutral Interstellar Medium: Absorption Spectroscopy of Galactic Hydrogen

    NASA Astrophysics Data System (ADS)

    Murray, Claire Elizabeth

    The formation of stars and evolution of galaxies depends on the cycle of interstellar matter between supernova-expelled plasma and molecule-rich gas. At the center of this cycle is multiphase neutral hydrogen (HI), whose physical conditions provide key ingredients to theoretical models. However, constraints for HI properties require measurements of gas emission and absorption which have been severely limited by previous observational capabilities. In this thesis, I present the largest survey of Galactic HI absorption ever undertaken with the Karl G. Jansky Very Large Array (VLA). The survey, 21 cm Spectral Line Observations of Neutral Gas with the VLA (21-SPONGE), is a statistical study of HI in all phases using direct absorption measurements. Leveraging novel calibration techniques, I demonstrate the capability of the VLA to detect a significant sample of 21 cm absorption lines from warm, diffuse HI. To maximize observational sensitivity, I stack the 21-SPONGE spectra and detect a pervasive signature of the warm neutral medium in absorption. The inferred excitation (or spin) temperature is consistent with existing estimates, yet higher than predictions from theoretical models of collisional HI excitation. This suggests that radiative feedback via resonant scattering of Lyalpha photons, known as the Wouthuysen-Field effect, is influential with important implications for cosmological 21 cm observations. Next, I compare 21-SPONGE with synthetic HI spectra from 3D numerical simulations using a new, objective decomposition and radiative transfer tool. I quantify the recovery of HI structures and their properties by Gaussian-fitted 21 cm spectral lines for the first time. I find that 21 cm absorption line shapes are sensitive to simulated physics, and demonstrate that my analysis method is a powerful tool for diagnosing neutral ISM conditions. Finally, I compare properties inferred from synthetic spectra with "true" simulation results to construct a bias correction function for estimating HI properties. I apply this correction to the mass distribution of HI as a function of temperature from 21-SPONGE, and find a significant fraction of thermally unstable gas. This confirms that non-steady radiative and dynamical processes, such as turbulence and supernovae, have a strong influence on the thermodynamic state of the ISM.

  2. The efficacy of topical human amniotic membrane-mesenchymal stem cell-conditioned medium (hAMMSC-CM) and a mixture of topical hAMMSC-CM + vitamin C and hAMMSC-CM + vitamin E on chronic plantar ulcers in leprosy:a randomized control trial.

    PubMed

    Prakoeswa, C R S; Natallya, F R; Harnindya, D; Thohiroh, A; Oktaviyanti, R N; Pratiwi, K D; Rubianti, M A; Yogatri, B; Primasari, P I; Herwanto, N; Alinda, M D; Kusumaputra, B H; Astari, L; Listiawan, M Y; Agusni, I; Rantam, F A

    2018-05-10

    Healing of chronic plantar ulcers in leprosy (CPUL) typically takes a long time due to impaired neurological function, thereby reducing the levels of growth factors and cytokines. Cytokines can be found in metabolite products from amniotic membrane stem cells. Chronic ulcers are frequently characterized by high levels of reactive oxygen species. Vitamin E (α-tocopherol) is widely used in skin lesions, owing to its antioxidant and anti-inflammatory properties. Vitamin C also has antioxidant, anti-inflammatory, and collagen synthesis properties which are useful in wound healing. Herein, we compared the effects of topical human amniotic membrane-mesenchymal stem cell-conditioned medium (hAMMSC-CM) alone and with vitamins C and E on healing of CPUL. In this randomized controlled trial, topical agents were applied every 3 days for up to 8 weeks. Ulcer size, side-effects, and possible complications were monitored weekly. Healing percentage increased each week in all groups. Mean difference in ulcer size was highest in the hAMMSC-CM + vitamin E group, implying better progress of wound healing. There were no side-effects or complications. hAMMSC-CM + vitamin E is best for healing of CPUL.

  3. IL-1β produced by aggressive breast cancer cells is one of the factors that dictate their interactions with mesenchymal stem cells through chemokine production.

    PubMed

    Escobar, Pauline; Bouclier, Céline; Serret, Julien; Bièche, Ivan; Brigitte, Madly; Caicedo, Andres; Sanchez, Elodie; Vacher, Sophie; Vignais, Marie-Luce; Bourin, Philippe; Geneviève, David; Molina, Franck; Jorgensen, Christian; Lazennec, Gwendal

    2015-10-06

    The aim of this work was to understand whether the nature of breast cancer cells could modify the nature of the dialog of mesenchymal stem cells (MSCs) with cancer cells. By treating MSCs with the conditioned medium of metastatic Estrogen-receptor (ER)-negative MDA-MB-231, or non-metastatic ER-positive MCF-7 breast cancer cells, we observed that a number of chemokines were produced at higher levels by MSCs treated with MDA-MB-231 conditioned medium (CM). MDA-MB-231 cells were able to induce NF-κB signaling in MSC cells. This was shown by the use of a NF-kB chemical inhibitor or an IκB dominant negative mutant, nuclear translocation of p65 and induction of NF-κB signature. Our results suggest that MDA-MB-231 cells exert their effects on MSCs through the secretion of IL-1β, that activates MSCs and induces the same chemokines as the MDA-MB-231CM. In addition, inhibition of IL-1β secretion in the MDA-MB-231 cells reduces the induced production of a panel of chemokines by MSCs, as well the motility of MDA-MB-231 cells. Our data suggest that aggressive breast cancer cells secrete IL-1β, which increases the production of chemokines by MSCs.

  4. IL-1β produced by aggressive breast cancer cells is one of the factors that dictate their interactions with mesenchymal stem cells through chemokine production

    PubMed Central

    Serret, Julien; Bièche, Ivan; Brigitte, Madly; Caicedo, Andres; Sanchez, Elodie; Vacher, Sophie; Vignais, Marie-Luce; Bourin, Philippe; Geneviève, David; Molina, Franck; Jorgensen, Christian; Lazennec, Gwendal

    2015-01-01

    The aim of this work was to understand whether the nature of breast cancer cells could modify the nature of the dialog of mesenchymal stem cells (MSCs) with cancer cells. By treating MSCs with the conditioned medium of metastatic Estrogen-receptor (ER)-negative MDA-MB-231, or non-metastatic ER-positive MCF-7 breast cancer cells, we observed that a number of chemokines were produced at higher levels by MSCs treated with MDA-MB-231 conditioned medium (CM). MDA-MB-231 cells were able to induce NF-κB signaling in MSC cells. This was shown by the use of a NF-kB chemical inhibitor or an IκB dominant negative mutant, nuclear translocation of p65 and induction of NF-κB signature. Our results suggest that MDA-MB-231 cells exert their effects on MSCs through the secretion of IL-1β, that activates MSCs and induces the same chemokines as the MDA-MB-231CM. In addition, inhibition of IL-1β secretion in the MDA-MB-231 cells reduces the induced production of a panel of chemokines by MSCs, as well the motility of MDA-MB-231 cells. Our data suggest that aggressive breast cancer cells secrete IL-1β, which increases the production of chemokines by MSCs. PMID:26362269

  5. Direct Electric Current Treatment under Physiologic Saline Conditions Kills Staphylococcus epidermidis Biofilms via Electrolytic Generation of Hypochlorous Acid

    PubMed Central

    Sandvik, Elizabeth L.; McLeod, Bruce R.; Parker, Albert E.; Stewart, Philip S.

    2013-01-01

    The purpose of this study was to investigate the mechanism by which a direct electrical current reduced the viability of Staphylococcus epidermidis biofilms in conjunction with ciprofloxacin at physiologic saline conditions meant to approximate those in an infected artificial joint. Biofilms grown in CDC biofilm reactors were exposed to current for 24 hours in 1/10th strength tryptic soy broth containing 9 g/L total NaCl. Dose-dependent log reductions up to 6.7 log10 CFU/cm2 were observed with the application of direct current at all four levels (0.7 to 1.8 mA/cm2) both in the presence and absence of ciprofloxacin. There were no significant differences in log reductions for wells with ciprofloxacin compared to those without at the same current levels. When current exposures were repeated without biofilm or organics in the medium, significant generation of free chlorine was measured. Free chlorine doses equivalent to the 24 hour endpoint concentration for each current level were shown to mimic killing achieved by current application. Current exposure (1.8 mA/cm2) in medium lacking chloride and amended with sulfate, nitrate, or phosphate as alternative electrolytes produced diminished kills of 3, 2, and 0 log reduction, respectively. Direct current also killed Pseudomonas aeruginosa biofilms when NaCl was present. Together these results indicate that electrolysis reactions generating hypochlorous acid from chloride are likely a main contributor to the efficacy of direct current application. A physiologically relevant NaCl concentration is thus a critical parameter in experimental design if direct current is to be investigated for in vivo medical applications. PMID:23390518

  6. Direct electric current treatment under physiologic saline conditions kills Staphylococcus epidermidis biofilms via electrolytic generation of hypochlorous acid.

    PubMed

    Sandvik, Elizabeth L; McLeod, Bruce R; Parker, Albert E; Stewart, Philip S

    2013-01-01

    The purpose of this study was to investigate the mechanism by which a direct electrical current reduced the viability of Staphylococcus epidermidis biofilms in conjunction with ciprofloxacin at physiologic saline conditions meant to approximate those in an infected artificial joint. Biofilms grown in CDC biofilm reactors were exposed to current for 24 hours in 1/10(th) strength tryptic soy broth containing 9 g/L total NaCl. Dose-dependent log reductions up to 6.7 log(10) CFU/cm(2) were observed with the application of direct current at all four levels (0.7 to 1.8 mA/cm(2)) both in the presence and absence of ciprofloxacin. There were no significant differences in log reductions for wells with ciprofloxacin compared to those without at the same current levels. When current exposures were repeated without biofilm or organics in the medium, significant generation of free chlorine was measured. Free chlorine doses equivalent to the 24 hour endpoint concentration for each current level were shown to mimic killing achieved by current application. Current exposure (1.8 mA/cm(2)) in medium lacking chloride and amended with sulfate, nitrate, or phosphate as alternative electrolytes produced diminished kills of 3, 2, and 0 log reduction, respectively. Direct current also killed Pseudomonas aeruginosa biofilms when NaCl was present. Together these results indicate that electrolysis reactions generating hypochlorous acid from chloride are likely a main contributor to the efficacy of direct current application. A physiologically relevant NaCl concentration is thus a critical parameter in experimental design if direct current is to be investigated for in vivo medical applications.

  7. Differences and effects of medium and large adult cuffs on blood pressure readings in individuals with muscular arms.

    PubMed

    Fonseca-Reyes, Salvador; Fajardo-Flores, Ismael; Montes-Casillas, Mayra; Forsyth-Macquarrie, Avril

    2009-08-01

    This study analyzed systolic and diastolic blood pressure (SBP and DBP) reading differences in individuals with muscular arms, using medium and large adult cuffs. Resting blood pressures (BPs) were measured in bodybuilders competing at the Mexican National Bodybuilding and Fitness Championship. The means of two bilateral simultaneous arm BP measurements were obtained using two different cuff sizes, 12 cm (medium adult) and 16 cm (large adult). A total of 193 bodybuilders completed the measurements. With an arm circumference greater than 33 cm, the SBP and DBP average taken with the medium adult cuff was higher than that obtained with the large adult cuff, 8.2+/-10.6 and 1.6+/-7.4 mmHg, respectively; however, a significant difference was observed only with the SBP. With the medium adult cuff, 48 of 144 individuals showed SBP at least 140 mmHg, whereas with the large adult cuff there were only 17 of 144 individuals. In those participants with an arm circumference less than 33 cm, the BP was nonsignificantly lower with the large cuff, -4.24+/-9.2 and -2.24+/-5.4 mmHg for the SBP and DBP, respectively. Incorrect cuffing of a muscular arm leads to significant errors in the measurement of SBP and DBP, similar to that observed in miscuffing of nonmuscular arms.

  8. Effect of torrefaction process on the coconut shell energy content for solid fuel

    NASA Astrophysics Data System (ADS)

    Irawan, Anton; Latifah Upe, S.; Meity Dwi I., P.

    2017-03-01

    Indonesia was one of largest coconut producers in the world with an average coconut production of 3 million tons per year and an estimated coconut shell waste were produced 360 thousand tons per year. Certainly, Coconut shell produced in large numbers require initial processing to be saved in the long term with stabilized quality. Quality of coconut shell can be maintained by changing the characteristics of the properties of coconut shell from easily absorbed water (hydrophilic) to difficult absorbed water (hydrophobia) as well as reduce the smoke of burning through torrefaction. Torrefaction technology carried out the biomass at a temperature of 200-300°C. The goal of this research was to observe the effect of operating conditions of torrefaction and the size of a coconut shell to the quality of coconut shell as a solid fuel which had high quality and low environmental impact. The variables in this study was the size of coconut shell (1.5 cm, 3 cm, and 4 cm), temperature (250°C, 300°C and 350°C) and torrefaction holding time (15, 30, and 45 minutes). Fresh coconut shell will be analyzed using proximate, ultimate analysis, and calorific value to know the initial condition. Torrefaction product will also be analyzed by proximate analysis and heating value. The highest calorific value was obtained on the size of coconut shell medium (3 cm) with operating conditions at a temperature of 350°C and torrefaction holding time 30 minutes at 7635 kcal /kg with the increasing percentage in calorific value 40.76%, fixed carbon 82.73%, and the volatile matter content 10.88%. But that condition of the torrefaction product has produced the low mass yield around 31%. The optimum conditions were at temperature 250°C, torrefaction holding time 30 minutes, and coconut shell size 1.5 cm.

  9. 78 FR 50346 - Atlantic Highly Migratory Species; Atlantic Bluefin Tuna Fisheries

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-08-19

    ... large medium or giant BFT to three large medium or giant BFT for the September, October through November... retention limit of one large medium or giant BFT (measuring 73 inches (185 cm) curved fork length (CFL) or... one large medium or giant BFT as follows: Two large medium or giant BFT for January (76 FR 76900...

  10. 3D Printed Structures Filled with Carbon Fibers and Functionalized with Mesenchymal Stem Cell Conditioned Media as In Vitro Cell Niches for Promoting Chondrogenesis.

    PubMed

    García-Ruíz, Josefa Predestinación; Díaz Lantada, Andrés

    2017-12-24

    In this study, we present a novel approach towards the straightforward, rapid, and low-cost development of biomimetic composite scaffolds for tissue engineering strategies. The system is based on the additive manufacture of a computer-designed lattice structure or framework, into which carbon fibers are subsequently knitted or incorporated. The 3D-printed lattice structure acts as support and the knitted carbon fibers perform as driving elements for promoting cell colonization of the three-dimensional construct. A human mesenchymal stem cell (h-MSC) conditioned medium (CM) is also used for improving the scaffold's response and promoting cell adhesion, proliferation, and viability. Cell culture results-in which scaffolds become buried in collagen type II-provide relevant information regarding the viability of the composite scaffolds used and the prospective applications of the proposed approach. In fact, the advanced composite scaffold developed, together with the conditioned medium functionalization, constitutes a biomimetic stem cell niche with clear potential, not just for tendon and ligament repair, but also for cartilage and endochondral bone formation and regeneration strategies.

  11. In vitro cultivation of anisakis simplex: pepsin increases survival and moulting from fourth larval to adult stage.

    PubMed

    Iglesias, L; Valero, A; Benítez, R; Adroher, F J

    2001-09-01

    This paper describes the in vitro cultivation of the 3rd-larval stage (L3) of Anisakis simplex to adulthood in a much simpler and easier to prepare medium than those described to date. The adult males obtained are between 3.8 and 6.5 cm long and the females between 4.5 and 8.0 cm. Some individually cultivated females laid eggs which had an average size of 44.4 x 50.5 microm. The culture conditions were as follows: medium RPMI-1640 supplemented with 20% heat-inactivated fetal bovine serum and 1% commercial pepsin, at pH 4.0 and a temperature of 37 degrees C, and in air atmosphere with 5% CO2. The pepsin was found to be the key to the success of the culture. The average survival of the worms in the culture increased from 50 to 88 days, due to the fact that the survival of the adults practically doubled (increasing by 1.9 times). Furthermore, the number of worms that completed the 4th moulting (M4) increased by 4.2 times, from 22.9 to 95.6%. This culture medium may facilitate, due to its simplicity, the study of anisakids, or at least of A. simplex, constituting another step towards achieving a complete in vitro life-cycle for these parasites.

  12. Establishment of an in vitro micropropagation protocol for Boscia senegalensis (Pers.) Lam. ex Poir.

    PubMed Central

    Khalafalla, Mutasim M.; Daffalla, Hussien M.; Abdellatef, Eltayb; Agabna, Elsadig; El-Shemy, Hany A.

    2011-01-01

    This report describes in vitro micropropagation of Boscia senegalensis, so-called famine foods, that helped the people in Darfur and Kordofan, Sudan survive during the 1984–1985 famine. Four types of explants prepared from green mature zygotic embryos were cultured on Murashige and Skoog (MS) medium augmented with 1–5 mg/L 6-benzyladenine (BA). The highest number of shoots per explant (14.3±0.9) was achieved on MS medium supplemented with 3 mg/L BA, while the highest shoot length [(3.5±0.4) cm] was obtained with 1 mg/L BA. The shoot cluster, when subcultured to its same medium, significantly increased the rate of shoot multiplication by the end of the third subculture. The maximum mean number of shoots per explant (86.5±3.6) was produced after three multiplication cycles on 3 mg/L BA-supplemented medium. In vitro induced shoots were excised and rooted on half strength MS medium fortified with 0.25 mg/L indole-3-butyric acid (IBA) to obtain complete plantlets. B. senegalensis-regenerated plantlets obtained in vitro for the first time, were hardened and 95% survived under greenhouse conditions. PMID:21462387

  13. Establishment of an in vitro micropropagation protocol for Boscia senegalensis (Pers.) Lam. ex Poir.

    PubMed

    Khalafalla, Mutasim M; Daffalla, Hussien M; Abdellatef, Eltayb; Agabna, Elsadig; El-Shemy, Hany A

    2011-04-01

    This report describes in vitro micropropagation of Boscia senegalensis, so-called famine foods, that helped the people in Darfur and Kordofan, Sudan survive during the 1984-1985 famine. Four types of explants prepared from green mature zygotic embryos were cultured on Murashige and Skoog (MS) medium augmented with 1-5 mg/L 6-benzyladenine (BA). The highest number of shoots per explant (14.3±0.9) was achieved on MS medium supplemented with 3 mg/L BA, while the highest shoot length [(3.5±0.4) cm] was obtained with 1 mg/L BA. The shoot cluster, when subcultured to its same medium, significantly increased the rate of shoot multiplication by the end of the third subculture. The maximum mean number of shoots per explant (86.5±3.6) was produced after three multiplication cycles on 3 mg/L BA-supplemented medium. In vitro induced shoots were excised and rooted on half strength MS medium fortified with 0.25 mg/L indole-3-butyric acid (IBA) to obtain complete plantlets. B. senegalensis-regenerated plantlets obtained in vitro for the first time, were hardened and 95% survived under greenhouse conditions.

  14. Abscisic Acid Participates in the Control of Cell Cycle Initiation Through Heme Homeostasis in the Unicellular Red Alga Cyanidioschyzon merolae.

    PubMed

    Kobayashi, Yuki; Ando, Hiroyuki; Hanaoka, Mitsumasa; Tanaka, Kan

    2016-05-01

    ABA is a phytohormone that is synthesized in response to abiotic stresses and other environmental changes, inducing various physiological responses. While ABA has been found in unicellular photosynthetic organisms, such as cyanobacteria and eukaryotic algae, its function in these organisms is poorly understood. Here, we found that ABA accumulated in the unicellular red alga Cyanidioschyzon merolae under conditions of salt stress and that the cell cycle G1/S transition was inhibited when ABA was added to the culture medium. A gene encoding heme-scavenging tryptophan-rich sensory protein-related protein (CmTSPO; CMS231C) was positively regulated by ABA, as in Arabidopsis, and CmTSPO bound heme in vitro. The intracellular content of total heme was increased by addition of ABA, but unfettered heme decreased, presumably due to scavenging by CmTSPO. The inhibition of DNA replication by ABA was negated by addition of heme to the culture medium. Thus, we propose a regulatory role for ABA and heme in algal cell cycle initiation. Finally, we found that a C. merolae mutant that is defective in ABA production was more susceptible to salt stress, indicating the importance of ABA to stress resistance in red algae. © The Author 2016. Published by Oxford University Press on behalf of Japanese Society of Plant Physiologists. All rights reserved. For permissions, please email: journals.permissions@oup.com.

  15. Soda-Anthraquinone Durian (Durio Zibethinus Murr.) Rind Linerboard and Corrugated Medium Paper: A Preliminary Test

    NASA Astrophysics Data System (ADS)

    Rizal Masrol, Shaiful; Irwan Ibrahim, Mohd Halim; Adnan, Sharmiza; Mubarak Sa'adon, Amir; Ika Sukarno, Khairil; Fadrol Hisham Yusoff, Mohd

    2017-08-01

    A preliminary test was conducted to investigate the characteristics of linerboard and corrugated medium paper made from durian rind waste. Naturally dried durian rinds were pulped according to Soda-Anthraquinone (Soda-AQ) pulping process with a condition of 20% active alkali, 0.1% AQ, 7:1 liquor to material ratio, 120 minutes cooking time and 170°C cooking temperature. The linerboard and corrugated medium paper with a basis weight of 120 gsm were prepared and evaluated according to Malaysian International Organization for Standardization (MS ISO) and Technical Association of the Pulp and Paper Industry (TAPPI). The results indicate that the characteristics of durian rind linerboard are comparable with other wood or non-wood based paper and current commercial paper. However, low CMT value for corrugated medium and water absorptiveness quality for linerboard could be improved in future. Based on the bulk density (0.672 g/cm3), burst index (3.12 kPa.m2/g) and RCT (2.00 N.m2/g), the durian rind has shown a good potential and suitable as an alternative raw material source for linerboard industry.

  16. Self-renewal of human embryonic stem cells requires insulin-like growth factor-1 receptor and ERBB2 receptor signaling

    PubMed Central

    Wang, Linlin; Schulz, Thomas C.; Sherrer, Eric S.; Dauphin, Derek S.; Shin, Soojung; Nelson, Angelique M.; Ware, Carol B.; Zhan, Mei; Song, Chao-Zhong; Chen, Xiaoji; Brimble, Sandii N.; McLean, Amanda; Galeano, Maria J.; Uhl, Elizabeth W.; D'Amour, Kevin A.; Chesnut, Jonathan D.; Rao, Mahendra S.

    2007-01-01

    Despite progress in developing defined conditions for human embryonic stem cell (hESC) cultures, little is known about the cell-surface receptors that are activated under conditions supportive of hESC self-renewal. A simultaneous interrogation of 42 receptor tyrosine kinases (RTKs) in hESCs following stimulation with mouse embryonic fibroblast (MEF) conditioned medium (CM) revealed rapid and prominent tyrosine phosphorylation of insulin receptor (IR) and insulin-like growth factor-1 receptor (IGF1R); less prominent tyrosine phosphorylation of epidermal growth factor receptor (EGFR) family members, including ERBB2 and ERBB3; and trace phosphorylation of fibroblast growth factor receptors. Intense IGF1R and IR phosphorylation occurred in the absence of MEF conditioning (NCM) and was attributable to high concentrations of insulin in the proprietary KnockOut Serum Replacer (KSR). Inhibition of IGF1R using a blocking antibody or lentivirus-delivered shRNA reduced hESC self-renewal and promoted differentiation, while disruption of ERBB2 signaling with the selective inhibitor AG825 severely inhibited hESC proliferation and promoted apoptosis. A simple defined medium containing an IGF1 analog, heregulin-1β (a ligand for ERBB2/ERBB3), fibroblast growth factor-2 (FGF2), and activin A supported long-term growth of multiple hESC lines. These studies identify previously unappreciated RTKs that support hESC proliferation and self-renewal, and provide a rationally designed medium for the growth and maintenance of pluripotent hESCs. PMID:17761519

  17. Prolonged swimming performance of northern squawfish

    USGS Publications Warehouse

    Mesa, Matthew G.; Olson, Todd M.

    1993-01-01

    We determined the prolonged swimming performance of two size-classes of northern squawfish Ptychocheilus oregonensis at 12 and 18°C. The percentage of fish fatigued was positively related to water velocity and best described by an exponential model. At 12°C, the velocity at which 50% of the fish fatigued (FV50) was estimated to be 2.91 fork lengths per second (FL/s; 100 cm/s) for medium-sized fish (30–39 cm) and 2.45 FL/s (104 cm/s) for large fish (40–49 cm). At 18°C, estimated FV50 was 3.12 FL/s (107 cm/s) for medium fish and 2.65 FL/s (112 cm/s) for large fish. Rate of change in percent fatigue was affected by fish size and water temperature. Large fish fatigued at a higher rate than medium-sized fish; all fish fatigued faster at 12 than at 18°C. The mean times to fatigue at velocities of 102–115 cm/s ranged from 14 to 28 min and were not affected by fish size or water temperature. Our results indicate that water velocities from 100 to 130 cm/s may exclude or reduce predation by northern squawfish around juvenile salmonid bypass outfalls at Columbia River dams, at least during certain times of the year. We recommend that construction or modification of juvenile salmonid bypass facilities place the outfall in an area of high water velocity and distant from eddies, submerged cover, and littoral areas.

  18. Breakup and fusion cross sections of the 6Li nucleus with targets of mass A = 58, 144 and 208

    NASA Astrophysics Data System (ADS)

    Mukeru, B.; Rampho, G. J.; Lekala, M. L.

    2018-04-01

    We use the continuum discretized coupled channels method to investigate the effects of continuum-continuum coupling on the breakup and fusion cross sections of the weakly bound 6Li nucleus with the 58Ni, 144Sm and 208Pb nuclear targets. The cross sections were analyzed at incident energies E cm below, close to and above the Coulomb barrier V B. We found that for the medium and heavy targets, the breakup cross sections are enhanced at energies below the Coulomb barrier (E cm/V B ≤ 0.8) owing to these couplings. For the lighter target, relatively small enhancement of the breakup cross sections appear at energies well below the barrier (E cm/V B ≤ 0.6). At energies E cm/V B > 0.8 for medium and heavy targets, and E cm/V B > 0.6 for the light target, the continuum-continuum couplings substantially suppress the breakup cross sections. On the other hand, the fusion cross sections are enhanced at energies E cm/V B < 1.4, E cm/V B < 1.2 and E cm/V B < 0.8 for the light, medium and heavy target, respectively. The enhancement decreases as the target mass increases. Above the indicated respective energies, these couplings suppress the fusion cross sections. We also compared the breakup and fusion cross sections, and found that below the barrier, the breakup cross sections are more dominant regardless of whether continuum-continuum couplings are included.

  19. Probing the Intergalactic Medium with Ly α and 21 cm Fluctuations

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

    Heneka, Caroline; Cooray, Asantha; Feng, Chang

    2017-10-10

    We study 21 cm and Ly α fluctuations, as well as H α , while distinguishing between Ly α emission of galactic, diffuse, and scattered intergalactic medium (IGM) origin. Cross-correlation information about the state of the IGM is obtained, testing neutral versus ionized medium cases with different tracers in a seminumerical simulation setup. In order to pave the way toward constraints on reionization history and modeling beyond power spectrum information, we explore parameter dependencies of the cross-power signal between 21 cm and Ly α , which displays a characteristic morphology and a turnover from negative to positive correlation at scalesmore » of a couple Mpc{sup −1}. In a proof of concept for the extraction of further information on the state of the IGM using different tracers, we demonstrate the use of the 21 cm and H α cross-correlation signal to determine the relative strength of galactic and IGM emission in Ly α . We conclude by showing the detectability of the 21 cm and Ly α cross-correlation signal over more than one decade in scale at high signal-to-noise ratio for upcoming probes like SKA and the proposed all-sky intensity mapping satellites SPHEREx and CDIM, while also including the Ly α damping tail and 21 cm foreground avoidance in the modeling.« less

  20. Dosimetric evaluation of IMRT plan for homogenous and inhomogeneous medium using AAPM TG-119 protocol

    NASA Astrophysics Data System (ADS)

    Fatimah, L. A. N.; Wibowo, W. E.; Pawiro, S. A.

    2017-05-01

    The American Association of Physicists in Medicine (AAPM) TG-119 protocol has been applied for dose verification in IMRT technique. However, some criteria in the protocol need to be verified for inhomogeneous medium and small volume targets. Hence, the purpose of this study was to verify the assessment criteria of dose verification in AAPM TG-119 for inhomogeneous medium and small volume targets. The work has been conducted by dose verification for homogeneous (phantom A) and inhomogeneous phantoms (phantom B and C) on two geometrical targets: C-shape and circular targets. The targets were simulated using 7 static dMLC IMRT fields at two different depths of 5 g/cm2 and 10 g/cm2. The dose optimisation and calculation were done by using Pinnacle3 for 6 MV photons beam. The planning objectives were set according to AAPM TG-119 parameters. The plan analysis was conducted by Conformity Index and Homogeneity Index. The point dose measurements were conducted with Exradin A16, Semiflex 0.125cc, and Gafchromic EBT3. The plan results show that CI for C-shape target is in the range of 0.710-0.999 at 10 g/cm2 depth and 0.691-1.613 at 5 g/cm2. In addition, HI for C-shape and circular were in the range of 6.3%-58.7% and 5.4%-87.1% for 10 g/cm2 depth. The measurement results show that the dose measurement at inhomogeneous medium and small volume targets are much lower than the criteria in AAPM TG-119. In conclusion, the criteria in the AAPM TG-119 cannot be fully implemented for inhomogeneous medium and small volume targets.

  1. Volatile metabolite production of spoilage micro-organisms on a mixed-lettuce agar during storage at 7 degrees C in air and low oxygen atmosphere.

    PubMed

    Ragaert, P; Devlieghere, F; Devuyst, E; Dewulf, J; Van Langenhove, H; Debevere, J

    2006-11-01

    This paper describes the volatile metabolite production of spoilage bacteria (Pantoea agglomerans and Rahnella aquatilis) and spoilage yeasts (Pichia fermentans and Cryptococcus laurentii), previously isolated from mixed lettuce, on a simulation medium of shredded mixed lettuce (mixed-lettuce agar) both under air conditions and modified atmosphere (MA)-conditions at 7 degrees C. These latter conditions simulated the equilibrium modified atmosphere packaging, which is used to extend the shelf-life of shredded mixed lettuce. Besides volatile metabolites, organic acid metabolites and consumption of sugars were measured. Microbiological growth on the mixed-lettuce agar resulted in metabolite production and consumption of sugars. Bacteria and yeasts produced a range of volatile organic compounds both under air conditions and MA-conditions: ethanol, ethyl acetate, 2-methyl-1-propanol, 2-methyl-1-butanol, 3-methyl-1-butanol, 2,3-butanedione, 3-methyl-1-pentanol, 1-butanol and 1-hexanol. Under MA-conditions, 2-methyl-1-butanol, 3-methyl-1-butanol and ethanol were the first compounds that were detected in the headspace as being produced by the inoculated micro-organisms. In the case of the yeast P. fermentans, production of these compounds was detected from a count of 5.0+/-0.1 log cfu/cm(2) with a fast increase when exceeding 6.0-6.5 log cfu/cm(2). Unlike P. fermentans, the yeast C. laurentii showed a slow metabolism under MA-conditions, compared to air conditions. In the case of the bacteria, production of 2-methyl-1-butanol and 3-methyl-1-butanol was detected starting from a count of 6.7+/-0.1 log cfu/cm(2) in the case of R. aquatilis and from a count of 7.1+/-0.4 log cfu/cm(2) in the case of P. agglomerans with a fast increase when exceeding 8 log cfu/cm(2). No production of ethanol by the bacteria under MA-conditions was detected in contradiction to air conditions. It could be concluded that, if these counts are reached on the cut surfaces of shredded mixed lettuce which are simulated by the mixed-lettuce agar, sensorial quality of shredded mixed lettuce could be influenced by the microbiological production of metabolites.

  2. Effect of recombinant bovine somatotropin (rbST) on cytoplasmic maturation of bovine oocytes and their developmental competence in vitro.

    PubMed

    Kuzmina, Tatjana I; Alm, Hannelore; Denisenko, Vitaly; Tuchscherer, Armin; Kanitz, Wilhelm; Torner, Helmut

    2007-04-01

    The objectives of this study were to evaluate the effects of recombinant bovine somatotropin (rbST) on the nuclear and cytoplasmic maturation of bovine oocytes and their further developmental competence to blastocysts in vitro. We analyzed the mitochondrial activity and concentration of intracellular stored calcium ([Ca(2+)](is)) in matured oocytes and the morphology and chromatin status of produced embryos after in vitro fertilization. Cumulus-oocyte complexes were incubated in TCM 199 containing 10% fetal calf serum (control medium 1: CM 1) or 10% estrus cow serum (control medium 2: CM 2). The culture medium of the treatment groups was modified by supplementation of the control medium with 10 ng/ml rbST (CM 1A and CM 2A), 10(6)/ml granulosa cells (CM 1B and CM 2B), or 10 ng/ml rbST plus 10(6)/ml granulosa cells (CM 1C and CM 2C). No differences were observed in the percentages of oocytes reaching metaphase II between the groups. However, the proportion of blastocysts was highest in treatment groups CM 1C and CM 2C (P<0.05). The type of serum did not alter the positive effect of rbST on the developmental competence of embryos. The fluorescence intensity of metabolically active mitochondria measured by intensity per oocyte (Em 570) after MitoTracker CMTM Ros Orange labeling was significantly increased in oocytes matured in the presence of 10 ng/ml rbST and granulosa cells (309.21 vs. 119.97 microA; P<0.01). In parallel, the concentration of [Ca(2+)](is) in oocytes, determined using fluorophore chlortetracycline, was significantly decreased (0.85 +/- 0.02 vs. 0.97 +/- 0.03 AU; P<0.05). Based on these results, we concluded that rbST, in interaction with granulosa cells stimulates the oxidative activity of ooplasmic mitochondria and decreases the content of [Ca(2+)](is) in oocytes. These facts support the hypothesis that somatotropin influences the developmental competence of bovine oocytes during maturation in vitro, and this effect can be modulated by granulosa cells.

  3. The efficacy of conditioned media of adipose-derived stem cells combined with ablative carbon dioxide fractional resurfacing for atrophic acne scars and skin rejuvenation.

    PubMed

    Zhou, Bing-Rong; Zhang, Ting; Bin Jameel, Afzaal Ahmed; Xu, Yang; Xu, Yan; Guo, Shi-Lei; Wang, Ying; Permatasari, Felicia; Luo, Dan

    2016-06-01

    To evaluate the effects of conditioned medium of adipose-derived stem cells (ADSC-CM) on efficacy and side effects after fractional carbon dioxide laser resurfacing (FxCR) when treating subjects with facial atrophic acne scars or with skin rejuvenation needs. Twenty-two subjects were enrolled in the study and divided into two groups. Nine subjects were included in skin rejuvenation group and thirteen subjects were included in acne scar group, and all subjects underwent three sessions of FxCR. ADSC-CM was applied on FxCR site of one randomly selected face side. Evaluations were done at baseline, 1 week after first treatment, and 1 month after each treatment. The outcome assessments included subjective satisfaction scale; blinded clinical assessment; and the biophysical parameters of roughness, elasticity, skin hydration, transepidermal water loss (TEWL), and the erythema and melanin index. Biopsies taken from one subject in skin rejuvenation group were analyzed using hematoxylin and eosin, Masson's Trichrome, and Gomori's aldehyde fuchsin staining. ADSC-CM combined with FxCR increased subject satisfaction, elasticity, skin hydration, and skin elasticity and decreased TEWL, roughness, and the melanin index in both acne scars and skin rejuvenation groups. Histologic analysis showed that ADSC-CM increased dermal collagen density, elastin density, and arranged them in order. ADSC-CM with FxCR is a good combination therapy for treating atrophic acne scars and skin rejuvenation. JSPH2012-082 - Registered 14 Feb 2012.

  4. Variation of Spirulina maxima biomass production in different depths of urea-used culture medium

    PubMed Central

    Affan, Md-Abu; Lee, Dae-Won; Al-Harbi, Salim Marzoog; Kim, Han-Jun; Abdulwassi, Najah Ibrahim; Heo, Soo-Jin; Oh, Chulhong; Park, Heung-Sik; Ma, Chae Woo; Lee, Hyeon-Yong; Kang, Do-Hyung

    2015-01-01

    Fewer studies have assessed the outdoor cultivation of Spirulina maxima compared with S. platensis, although the protein content of S. maxima is higher than S. platensis. Spirulina growth medium requires an increased amount of NaHCO3, Na2CO3, and NaNO3, which increases the production cost. Therefore, the current study used a low-cost but high-efficiency biomass production medium (Medium M-19) after testing 33 different media. The medium depth of 25 cm (group A) was sub-divided into A1 (50% cover with a black curtain (PolyMax, 12 oz ultra-blackout), A2 (25% cover), and A3 (no cover). Similarly the medium depths of 30 and 35 cm were categorized as groups B (B1, B2, and B3) and C (C1, C2, and C3), respectively, and the effects of depth and surface light availability on growth and biomass production were assessed. The highest biomass production was 2.05 g L-1 in group A2, which was significantly higher (p < 0.05) than that in all other groups and sub-groups. Spirulina maxima died in B1 and C1 on the fifth day of culture. The biochemical composition of the biomass obtained from A2 cultures, including protein, carbohydrate, lipid, moisture, and ash, was 56.59%, 14.42%, 0.94%, 5.03%, and 23.02%, respectively. Therefore, S. maxima could be grown outdoors with the highest efficiency in urea-enriched medium at a 25-cm medium depth with 25% surface cover or uncovered. PMID:26691456

  5. Variation of Spirulina maxima biomass production in different depths of urea-used culture medium.

    PubMed

    Affan, Md-Abu; Lee, Dae-Won; Al-Harbi, Salim Marzoog; Kim, Han-Jun; Abdulwassi, Najah Ibrahim; Heo, Soo-Jin; Oh, Chulhong; Park, Heung-Sik; Ma, Chae Woo; Lee, Hyeon-Yong; Kang, Do-Hyung

    2015-01-01

    Fewer studies have assessed the outdoor cultivation of Spirulina maxima compared with S. platensis, although the protein content of S. maxima is higher than S. platensis. Spirulina growth medium requires an increased amount of NaHCO3, Na2CO3, and NaNO3, which increases the production cost. Therefore, the current study used a low-cost but high-efficiency biomass production medium (Medium M-19) after testing 33 different media. The medium depth of 25 cm (group A) was sub-divided into A1 (50% cover with a black curtain (PolyMax, 12 oz ultra-blackout), A2 (25% cover), and A3 (no cover). Similarly the medium depths of 30 and 35 cm were categorized as groups B (B1, B2, and B3) and C (C1, C2, and C3), respectively, and the effects of depth and surface light availability on growth and biomass production were assessed. The highest biomass production was 2.05 g L-1 in group A2, which was significantly higher (p < 0.05) than that in all other groups and sub-groups. Spirulina maxima died in B1 and C1 on the fifth day of culture. The biochemical composition of the biomass obtained from A2 cultures, including protein, carbohydrate, lipid, moisture, and ash, was 56.59%, 14.42%, 0.94%, 5.03%, and 23.02%, respectively. Therefore, S. maxima could be grown outdoors with the highest efficiency in urea-enriched medium at a 25-cm medium depth with 25% surface cover or uncovered.

  6. Inbuilt potential of YEM medium and its constituents to generate Ag/Ag₂O nanoparticles.

    PubMed

    Yamal, G; Sharmila, P; Rao, K S; Pardha-Saradhi, P

    2013-01-01

    We discovered that Yeast Extract Mannitol (YEM) medium possessed immense potential to generate silver nanoparticles from AgNO3 upon autoclaving, which was evident from (i) alteration in color of the medium; (ii) peak at ∼410 nm in UV-Vis spectrum due to surface plasmon resonance specific to silver nanoparticles; and (iii) TEM investigations. TEM coupled with EDX confirmed that distinct nanoparticles were composed of silver. Yeast extract and mannitol were key components of YEM medium responsible for the formation of nanoparticles. PXRD analysis indicated crystalline geometry and Ag/Ag2O phases in nanoparticles generated with YEM medium, yeast extract and mannitol. Our investigations also revealed that both mannitol and yeast extract possessed potential to convert ∼80% of silver ions in 0.5 mM AgNO3 to nanoparticles, on autoclaving for 30 min at 121°C under a pressure of 1.06 kg/cm(2). Addition of filter sterilized AgNO3 under ambient conditions to pre-autoclaved YEM medium and yeast extract brought about color change due to the formation of silver nanoparticles, but required prolonged duration. In general, even after 72 h intensity of color was significantly less than that recorded following autoclaving. Silver nanoparticles formed at room temperature were more heterogeneous compared to that obtained upon autoclaving. In summary, our findings demonstrated that (i) YEM medium and its constituents promote synthesis of silver nanoparticles; and (ii) autoclaving enhances rapid synthesis of silver nanoparticles by YEM medium, yeast extract and mannitol.

  7. Aspect of ECMWF downscaled Regional Climate Modeling in simulating Indian summer monsoon rainfall and dependencies on lateral boundary conditions

    NASA Astrophysics Data System (ADS)

    Ghosh, Soumik; Bhatla, R.; Mall, R. K.; Srivastava, Prashant K.; Sahai, A. K.

    2018-03-01

    Climate model faces considerable difficulties in simulating the rainfall characteristics of southwest summer monsoon. In this study, the dynamical downscaling of European Centre for Medium-Range Weather Forecast's (ECMWF's) ERA-Interim (EIN15) has been utilized for the simulation of Indian summer monsoon (ISM) through the Regional Climate Model version 4.3 (RegCM-4.3) over the South Asia Co-Ordinated Regional Climate Downscaling EXperiment (CORDEX) domain. The complexities of model simulation over a particular terrain are generally influenced by factors such as complex topography, coastal boundary, and lack of unbiased initial and lateral boundary conditions. In order to overcome some of these limitations, the RegCM-4.3 is employed for simulating the rainfall characteristics over the complex topographical conditions. For reliable rainfall simulation, implementations of numerous lower boundary conditions are forced in the RegCM-4.3 with specific horizontal grid resolution of 50 km over South Asia CORDEX domain. The analysis is considered for 30 years of climatological simulation of rainfall, outgoing longwave radiation (OLR), mean sea level pressure (MSLP), and wind with different vertical levels over the specified region. The dependency of model simulation with the forcing of EIN15 initial and lateral boundary conditions is used to understand the impact of simulated rainfall characteristics during different phases of summer monsoon. The results obtained from this study are used to evaluate the activity of initial conditions of zonal wind circulation speed, which causes an increase in the uncertainty of regional model output over the region under investigation. Further, the results showed that the EIN15 zonal wind circulation lacks sufficient speed over the specified region in a particular time, which was carried forward by the RegCM output and leads to a disrupted regional simulation in the climate model.

  8. Glioma-secreted soluble factors stimulate microglial activation: The role of interleukin-1β and tumor necrosis factor-α.

    PubMed

    Hwang, Ji-Sun; Jung, Eun-Hye; Kwon, Mi-Youn; Han, Inn-Oc

    2016-09-15

    We aimed to elucidate the effect of soluble factors secreted by glioma on microglial activation. Conditioned medium (CM) from glioma cells, CRT-MG and C6, significantly induced nitric oxide (NO) production and stimulated the mRNA expression of inducible NO synthase (iNOS), interleukin (IL)-1beta, IL-6, tumor necrosis factor-alpha (TNF-α) and cyclooxygenase 2 (COX-2) in BV2 cells. Glioma CM stimulated p38 mitogen-activated protein kinase (MAPK) phosphorylation, and a p38 MAPK inhibitor, SB203580, suppressed CM-induced NO production in BV2 cells. In addition, CM stimulated nuclear factor-kappaB (NF-κB) DNA binding and transcriptional activity, which was repressed by SB203580. Gliomas displayed higher mRNA expression and release of TNF-α and IL-1β than primary astrocyte cells. Neutralization of TNF-α and IL-1β in C6-CM using a neutralizing antibody inhibited NO/iNOS expression in BV-2 cells. These results indicate potential contribution of diffusible tumor-derived factors to regulate microglial activation and subsequent tumor microenvironment. Copyright © 2016. Published by Elsevier B.V.

  9. Anomalous behavior in temporal evolution of ripple wavelength under medium energy Ar{sup +}-ion bombardment on Si: A case of initial wavelength selection

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

    Garg, Sandeep Kumar; Inter-University Accelerator Centre, Aruna Asaf Ali Marg, New Delhi 110 067; Cuerno, Rodolfo

    We have studied the early stage dynamics of ripple patterns on Si surfaces, in the fluence range of 1–3 × 10{sup 18} ions cm{sup −2}, as induced by medium energy Ar{sup +}-ion irradiation at room temperature. Under our experimental conditions, the ripple evolution is found to be in the linear regime, while a clear decreasing trend in the ripple wavelength is observed up to a certain time (fluence). Numerical simulations of a continuum model of ion-sputtered surfaces suggest that this anomalous behavior is due to the relaxation of the surface features of the experimental pristine surface during the initial stage of patternmore » formation. The observation of this hitherto unobserved behavior of the ripple wavelength seems to have been enabled by the use of medium energy ions, where the ripple wavelengths are found to be order(s) of magnitude larger than those at lower ion energies.« less

  10. Dust control by air-blocking shelves and dust collector-to-bailing airflow ratios for a surface mine drill shroud

    PubMed Central

    Zheng, Y.; Reed, W.R.; Potts, J.D.; Li, M.; Rider, J.P.

    2018-01-01

    The National Institute for Occupational Safety and Health (NIOSH) recently developed a series of validated models utilizing computational fluid dynamics (CFD) to study the effects of air-blocking shelves on airflows and respirable dust distribution associated with medium-sized surface blasthole drill shrouds as part of a dry dust collector system. Using validated CFD models, three different air-blocking shelves were included in the present study: a 15.2-cm (6-in.)-wide shelf; a 7.6-cm (3-in.)-wide shelf; and a 7.6-cm (3-in.)-wide shelf at four different shelf heights. In addition, the dust-collector-to-bailing airflow ratios of 1.75:1, 1.5:1, 1.25:1 and 1:1 were evaluated for the 15.2-cm (6-in.)-wide air-blocking shelf. This paper describes the methodology used to develop the CFD models. The effects of air-blocking shelves and dust collector-to-bailing airflow ratios were identified by the study, and problem regions were revealed under certain conditions.

  11. Localization of a small change in a multiple scattering environment without modeling of the actual medium.

    PubMed

    Rakotonarivo, S T; Walker, S C; Kuperman, W A; Roux, P

    2011-12-01

    A method to actively localize a small perturbation in a multiple scattering medium using a collection of remote acoustic sensors is presented. The approach requires only minimal modeling and no knowledge of the scatterer distribution and properties of the scattering medium and the perturbation. The medium is ensonified before and after a perturbation is introduced. The coherent difference between the measured signals then reveals all field components that have interacted with the perturbation. A simple single scatter filter (that ignores the presence of the medium scatterers) is matched to the earliest change of the coherent difference to localize the perturbation. Using a multi-source/receiver laboratory setup in air, the technique has been successfully tested with experimental data at frequencies varying from 30 to 60 kHz (wavelength ranging from 0.5 to 1 cm) for cm-scale scatterers in a scattering medium with a size two to five times bigger than its transport mean free path. © 2011 Acoustical Society of America

  12. Nutritional stress enhances cell viability of odontoblast-like cells subjected to low level laser irradiation

    NASA Astrophysics Data System (ADS)

    Tagliani, M. M.; Oliveira, C. F.; Lins, E. M. M.; Kurachi, C.; Hebling, J.; Bagnato, V. S.; de Souza Costa, C. A.

    2010-03-01

    In spite of knowing that cells under stress are biostimulated by low level laser (LLL) irradiation, the ideal condition of stress to different cell lines has not yet been established. Consequently, the aim of the present in vitro study was to evaluate the effects of a defined parameter of LLL irradiation applied on stressed odontoblast-like pulp cells (MDPC-23). The cells were seeded (12500 cells/cm2) in wells of 24-well plates using complete culture medium (DMEM) and incubated for 24 hours. Then, the DMEM was replaced by a new medium with low concentrations (nutritional stress condition) of fetal bovine serum (FBS) giving rise to the following experimental groups: G1: 2% FBS; G2: 5% FBS; and G3: 10% FBS. The cells were irradiated three times with LLL in specific parameters (808±3 nm, 100 mW, 1.5 J/cm2) every 24 hours. No irradiation was carried out in groups G4 (2% FBS-Control), G5 (5% FBS-Control), and G6 (10% FBS-Control). For all groups, the cell metabolism (MTT assay) and morphology (SEM) was evaluated. The experimental groups showed enhanced cell metabolism and normal cell morphology regardless of FBS concentration. A slight increase in the cell metabolism was observed only in group G2. It was concluded that cell nutritional stress caused by reducing the concentration of FBS to 5% is the most suitable method to assess the biostimulation of LLL irradiated MDPC-23 cells.

  13. Tracing Thermal Creep Through Granular Media

    NASA Astrophysics Data System (ADS)

    Steinpilz, Tobias; Teiser, Jens; Koester, Marc; Schywek, Mathias; Wurm, Gerhard

    2017-08-01

    A temperature gradient within a granular medium at low ambient pressure drives a gas flow through the medium by thermal creep. We measured the resulting air flow for a sample of glass beads with particle diameters between 290 μ m and 420 μ m for random close packing. Ambient pressure was varied between 1 Pa and 1000 Pa. The gas flow was quantified by means of tracer particles during parabolic flights. The flow varies systematically with pressure between 0.2 cm/s and 6 cm/s. The measured flow velocities are in quantitative agreement to model calculations that treat the granular medium as a collection of linear capillaries.

  14. An in vitro study of magnetic particle targeting in small blood vessels

    NASA Astrophysics Data System (ADS)

    Udrea, Laura Elena; Strachan, Norval J. C.; Bădescu, Vasile; Rotariu, Ovidiu

    2006-10-01

    The magnetic guidance and capture of particles inside the human body, via the circulatory system, is a novel method for the targeted delivery of drugs. This experimental study confirms in vitro that a dipolar capturing device, based on high-energy magnets with an active space of 8.7 cm × 10 cm × 10 cm, retains colloidal magnetic particles (MPs) (<30 nm) injected in the capillary tubes, where flow velocities are comparable to that encountered in the capillary beds of tumours (<0.5 cm s-1). The build-up of the deposition of the MPs was investigated using video imaging techniques that enabled continuous monitoring of the blocking of the vessel whilst simultaneously recording the colloid's flow rate. The parameters of practical importance (length of MP deposit, time of capillary blocking) were estimated and were found to be dependent on the initial fluid velocity, the MP concentration and the distance between the capillary tube and the polar magnetic pieces. Although the tube used in this experiment is larger (diameter = 0.75 mm, length = 100 mm) than that of real capillaries (diameter = 0.01 mm, length ~1.5 mm), the flow velocities chosen were similar to those encountered in the capillary beds of tumours and the length/diameter ratio was approximately equal (133 for the present set-up, 100-150 for real capillaries). In these circumstances and using the same magnetic field conditions (intensity, gradient) and MPs, there is close similarity with magnetic capture in a microscopic capillary system. Moreover, the macroscopic system permits analysis of the distribution of MPs in the active magnetic space, and consequently the maximum targetable volume. This study revealed that the capture of particles within the active space was strongly influenced by the gradient of the magnetic field and the flow velocity. Thus, when the magnetic field gradient had medium values (0.1-0.3 T cm-1) and the fluid velocity was small (0.15 cm s-1), the particles were captured in small, compact and stable deposits (L < 4 cm) and the time necessary for blocking of the capillary was <150 s. Doubling the value for the flow velocity did not influence significantly either the length of MP deposits nor the blocking time. However, lower gradients (<0.1 T cm-1) and larger velocities (0.3-0.9 cm s-1) result in the formation of larger deposits (4 cm < L < 10 cm) that are unstable at the beginning of the capture process. These large deposits do become stable given sufficient time for the deposition process to take place in conjunction with a decrease in the flow rate. As a consequence, the time necessary for blocking of the capillary increased up to 450 s. Decreasing the MP concentration from 0.02 g cm-3 to 0.005 g cm-3 decreased the deposit lengths by approximately 20% and doubled the values of the blocking time. The maximum targetable volume obtained by the present method is ~350 cm3, which corresponds to medium-sized tumours. The capillary vessels were blocked only for the situation that occurs for microcirculation within a tumour. This reduces the concentration of MPs trapped within the normal tissues, which occurs when using particles of micrometre size. This work showed the potential of using colloidal MPs and dipolar magnetic devices for treatment of human patients, when the affected sites are positioned at medium distances from the surface of the body (e.g. head, neck, breast, hands and legs).

  15. Influence of Silica Nanoparticles on Antioxidant Potential of Bacillus subtilis IMV B-7023

    NASA Astrophysics Data System (ADS)

    Skorochod, Iryna O.; Roy, Alla O.; Kurdish, Ivan K.

    2016-03-01

    It was found that if introduced into a nutrient medium of 0.05-1 g/L nano-SiO2, the oxidant activity (OA) of the culture medium (CM) of bacilli increased by 43.2-60.1 % and the antioxidant activity (AA) decreased by 4.5-11.8 %. SiO2 nanoparticles had different effects on antiradical activity (ARA) of the CM of Bacillus subtilis IMV B-7023. In particular, nano-SiO2 had no significant effect on the ability of the CM of bacilli to inactivate the 2.2-diphenyl-1-picrylhydrazyl (DPPH·) free radical. However, for the content of the nanomaterial of 0.01-1 g/L decreased hydroxyl radical scavenging in the CM of B. subtilis IMV B-7023 on 7.2-17.6 % compared with a control. Low doses of silica nanoparticles stimulated the reducing power of the CM of bacteria and then highly suppressed it.

  16. 76 FR 76900 - Atlantic Highly Migratory Species; Atlantic Bluefin Tuna Fisheries

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-12-09

    ... retention limit of one large medium or giant BFT (measuring 73 inches (185 cm) CFL) or greater per vessel... level of one large medium or giant BFT as follows: Two large medium or giant BFT for January (75 FR 79309, December 20, 2010); three large medium or giant BFT for June through August (76 FR 32086, June 3...

  17. 75 FR 79309 - Atlantic Highly Migratory Species; Atlantic Bluefin Tuna Fisheries

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-12-20

    ... limit of one large medium or giant BFT (measuring 73 inches (185 cm) CFL) or greater per vessel per day... adjusted the General category limit from the default level of one large medium or giant BFT as follows: Two large medium or giant BFT for January (74 FR 68709, December 29, 2009), and three large medium or giant...

  18. Non-thermal plasma modified growth and differentiation process of Capsicum annuum PP805 Godiva in in vitro conditions

    NASA Astrophysics Data System (ADS)

    Safari, Nasrin; Iranbakhsh, Alireza; Ardebili, Zahra Oraghi

    2017-05-01

    With the aim of evaluating the possible impacts of cold plasma on the structure and growth pattern of Capsicum annuum, the current study was carried out. The seeds were exposed to an argon-derived plasma (0.84 W cm-2 surface power densities) for 0, 1 or 2 minutes. Plasma-treated seeds were grown in the Murashige and Skoog (MS) medium or MS medium supplemented with BA and IAA. The presence of purple stems was recorded in plasma-treated plants grown in the medium supplemented with hormones. The recorded morphological differences were dependent on the exposure time of plasma treatments and/or the presence of hormones in the culture media. Plasma treatment of 1 minute had an improving effect on the shoot and root lengths as well as total leaf area, whereas plasma treatment of 2 minutes had an adverse effect. In contrast to the 1 minute treatment, plasma treatment of 2 minutes significantly impaired growth and hence reduced the total biomass. Alterations in stem diameter and differences in tissue patterns (especially in the vascular system) occurred, and were mainly dependent on the plasma exposure time and/or the presence of hormones. This is a first report on the effects of cold plasma on plant growth in in vitro conditions.

  19. Microfacies of mappable Archean biomats, Moodies Group, Barberton Greenstone Belt, South Africa

    NASA Astrophysics Data System (ADS)

    Gamper, Antonia; Heubeck, Christoph; Ohnemueller, Frank; Walsh, Maud

    2010-05-01

    The ca. 3.22 Ga-old Moodies Group, Barberton Mountain Land, South Africa, arguably includes the world's oldest regionally mappable biofacies. There, abundant smooth, wavy, domal or cuspate, interwoven or parallel-stratified laminae of isotopically light kerogen (Noffke et al. 2006) in shallow-water or coastal environments show a microtopography of several cm, deform cohesively, trap and bind grains, and were surficially rapidly silicified. In order to investigate the microfacies and habitat of these extensive biomats, we measured stratigraphic sections, sampled for petrography and composition, and documented sedimentary structures throughout. Seven stratigraphic sections allow the reconstruction of a coastal depositional system with an thickness of approx. 240 m along an > 11 km long outcrop belt. The system can be subdivided in (from base to top deepening) terrestrial coastal, low-angle shoreline, subtidal and shoreface facies. Biomats are most densely (mm- to cm-) spaced in the shoreface unit whereas they are least common in the basal terrestrial unit in which single-pebble trains and thin gravel conglomerates occur. Biomats (mean 4 mm thick) reach their greatest individual thickness (up to 0,8 cm) and dominate the spectrum of sedimentary structures in the subtidal unit where they form black, internally laminated chert bands. Most chert bands overlie lenses of elongate, well-sorted, coarse-grained sandstone but are in turn sharply overlain by medium- and fine-grained sandstone, suggesting cyclic current activity. Clustered or regularly spaced (sub-)vertical fluid escape structures penetrate and ductily deform densely spaced interwoven biomats. They occur most widely in the shoreface facies, show a mean height of 49 cm, are commonly offset horizontally, and reach up to 230 cm. The margins of several shallow (max. 1 m deep) and up to 8 m wide channels erosively truncate wrinkled biomats of the terrestrial coastal facies. Channel fill includes dominant medium- to coarse-grained sand and subordinate palm-sized planar (apparently brittle) biomat fragments mixed with granule and pebble lag at the channel base. Channel dimensions and erosivity may suggest a subaerial setting and imply that biomat growth either predated a temporary base-level drop or grew on land, presumably in a flat coastal setting. The micromorphology, large lateral extent, and depositional architecture of the biomats indicate a well-developed, adaptable, resistant microbial ecosystem along a medium-energy coast and offer a remarkable window in the conditions under which early life on Earth developed.

  20. Dicke’s Superradiance in Astrophysics. I. The 21 cm Line

    NASA Astrophysics Data System (ADS)

    Rajabi, Fereshteh; Houde, Martin

    2016-08-01

    We have applied the concept of superradiance introduced by Dicke in 1954 to astrophysics by extending the corresponding analysis to the magnetic dipole interaction characterizing the atomic hydrogen 21 cm line. Although it is unlikely that superradiance could take place in thermally relaxed regions and that the lack of observational evidence of masers for this transition reduces the probability of detecting superradiance, in situations where the conditions necessary for superradiance are met (close atomic spacing, high velocity coherence, population inversion, and long dephasing timescales compared to those related to coherent behavior), our results suggest that relatively low levels of population inversion over short astronomical length-scales (e.g., as compared to those required for maser amplification) can lead to the cooperative behavior required for superradiance in the interstellar medium. Given the results of our analysis, we expect the observational properties of 21 cm superradiance to be characterized by the emission of high-intensity, spatially compact, burst-like features potentially taking place over short periods ranging from minutes to days.

  1. Modulation of cathepsin G expression in severe atopic dermatitis following medium-dose UVA1 phototherapy

    PubMed Central

    Breuckmann, Frank; von Kobyletzki, Gregor; Avermaete, Annelies; Kreuter, Alexander; Altmeyer, Peter; Gambichler, Thilo

    2002-01-01

    Background During the last decade, medium-dose UVA1 phototherapy (50 J/cm2) has achieved great value within the treatment of severe atopic dermatitis (AD). The purpose of our study was to investigate to what extent UVA1 irradiation is able to modulate the status of protease activity by the use of a monoclonal antibody labeling cathepsin G. Methods In order to further elucidate the mechanisms by which medium-dose UVA1 irradiation leads to an improvement of skin status in patients with AD, biopsy specimens from 15 patients before and after treatment were analyzed immunohistochemically for proteolytic activation. Results Compared to lesional skin of patients with AD before UVA1 irradiation, the number of cells positive for cathepsin G within the dermal infiltrate decreased significantly after treatment. The decrease of cathepsin G+ cells was closely linked to a substantial clinical improvement in skin condition. Conclusions In summary, our findings demonstrated that medium-dose UVA1 irradiation leads to a modulation of the expression of cathepsin G in the dermal inflammatory infiltrate in patients with severe AD. Cathepsin G may attack laminin, proteoglycans, collagen I and insoluble fibronectin, to provoke proinflammatory events, to degrade the basement membrane, to destroy the tissue inhibitor of metalloproteinases and to increase the endothelial permeability. Therefore, its down-regulation by UVA1 phototherapy may induce the reduction of skin inflammation as well as improvement of the skin condition. PMID:12204095

  2. Investigation of groundwater in parts of Ndokwa District in Nigeria using geophysical logging and electrical resistivity methods: Implications for groundwater exploration

    NASA Astrophysics Data System (ADS)

    Anomohanran, Ochuko; Ofomola, Merrious Oviri; Okocha, Fredrick Ogochukwu

    2017-05-01

    Groundwater study involving the application of geophysical logging and vertical electrical sounding (VES) methods was carried out in parts of Ndokwa area of Delta State, Nigeria. The objective was to delineate the geological situation and the groundwater condition of the area. The geophysical logging of a drilled well and thirty VESs of the Schlumberger configuration were executed in this study using the Abem SAS 1000/4000 Terrameter. The result of the lithological study from the drilled well showed that the subsurface formation consist of lateritic topsoil, very fine sand, clayey fine sand, fine and medium grain sand, coarse sand, medium coarse sand and very coarse sand. The interpretation of the vertical electrical sounding data using a combination of curve matching and Win Resist computer iteration showed a close correlation with the well record. The result revealed the presence of four geoelectric layers with the aquifer identified to be in the fourth layer and having resistivity which ranged from 480 to 11,904 Ωm, while the depth ranged between 17.8 and 38.8 m. The analysis of the geophysical logging revealed that the average value of the electrical conductivity and the total dissolved solid of the groundwater in the aquifer were obtained as 229 μS/cm and 149 mg/cm3 respectively. These results indicate that the groundwater is within the permissible limit set by the Standard Organization of Nigeria for potable water which is 1000 μS/cm for electrical conductivity and 500 mg/cm3 for total dissolved solid. The fourth layer was therefore identified as the potential non conductive zone suitable for groundwater development in the study area.

  3. Wireless power transmission for biomedical implants: The role of near-zero threshold CMOS rectifiers.

    PubMed

    Mohammadi, Ali; Redoute, Jean-Michel; Yuce, Mehmet R

    2015-01-01

    Biomedical implants require an electronic power conditioning circuitry to provide a stable electrical power supply. The efficiency of wireless power transmission is strongly dependent on the power conditioning circuitry specifically the rectifier. A cross-connected CMOS bridge rectifier is implemented to demonstrate the impact of thresholds of rectifiers on wireless power transfer. The performance of the proposed rectifier is experimentally compared with a conventional Schottky diode full wave rectifier over 9 cm distance of air and tissue medium between the transmitter and receiver. The output voltage generated by the CMOS rectifier across a 1 KΩ resistive load is around twice as much as the Schottky rectifier.

  4. Validation of the Microlife WatchBP Home blood pressure device in pregnancy for medium and large arm circumferences.

    PubMed

    Clark, Katherine; Snowball, Olivia; Nzelu, Diane; Kay, Polly; Kametas, Nikos A

    2018-06-01

    The Microlife WatchBP Home automated blood pressure device was assessed for accuracy in pregnant women of medium (<32 cm) and large (≥32 cm) arm circumference. The British Hypertension Society validation protocol was modified for the purpose of this study to include women with arm circumference of less than 32 cm (N=51) and greater than or equal to 32 cm (N=46) as two separate arms. The device achieved an overall A/A grade for medium arm circumference and B/A grade for large arm circumference. The mean±SD device-observer difference was 1.7±6.2 and -0.4±4.4 for systolic and diastolic blood pressure, respectively, for medium arm circumference and 3.0±8.5 and 1.5±5.1, respectively, for large arm circumference. When all women with pre-eclampsia from both groups were pooled (N=23), the device achieved an overall grade of A/A with mean differences of 2.1±7.2 for systolic blood pressure and 1.0±5.6 for diastolic blood pressure. The Microlife WatchBP Home automated blood pressure device can be recommended for use in pregnant women of all gestations, including those with pre-eclampsia. However, caution is needed for women with large arm circumferences.

  5. Corticotrophin-releasing hormone and corticosterone impair development of preimplantation embryos by inducing oviductal cell apoptosis via activating the Fas system: an in vitro study.

    PubMed

    Tan, Xiu-Wen; Ji, Chang-Li; Zheng, Liang-Liang; Zhang, Jie; Yuan, Hong-Jie; Gong, Shuai; Zhu, Jiang; Tan, Jing-He

    2017-08-01

    What are the mechanisms by which corticotrophin-releasing hormone (CRH) and corticosterone impair the development of preimplantation embryos in the oviduct. CRH and corticosterone do not affect preimplantation embryos directly, but impair their development indirectly by triggering apoptosis of oviductal epithelial cells (OECs) through activation of the Fas system. Studies report that stress impairs embryo development with facilitated secretion of CRH and glucocorticoids. Although an in vivo study demonstrated that preimplantation stress impaired embryo development in conjunction with oviductal apoptosis and activation of the Fas system, whether CRH or glucocorticoids damage embryos directly or indirectly by way of oviductal cells remains to be clarified. Mice of Kunming strain, the generalized lymphoproliferative disorder (gld) mice with a germline mutation F273L in Fas ligand in a C57BL/6J genomic background and the wild-type C57BL/6J mice were used. Female mice were used 8-10 weeks after birth. While some female mice were killed 48 h after being injected with equine CG to collect oviducts and prepare OECs, others were killed to recover zygotes after mating with males following superovulation with eCG and hCG. The zygotes obtained were cultured with or without CRH or corticosterone (CRH/Cort) either in Chatot-Ziomek-Bavister (CZB) medium with or without OECs or in conditioned medium (CM) conditioned with OECs pretreated or not with CRH/Cort. Preimplantation development, levels of redox potential and apoptosis, and expression of CRH receptor 1 (CRHR1), glucocorticoid receptor (GR), Fas and 11β-hydroxysteroid dehydrogenase (HSD) were observed in embryos recovered at different times of in vitro culture. After culture of OECs with or without CRH/Cort, levels of redox potential and apoptosis, mRNA and protein expression of growth factors, and protein expression of CRHR1, GR and Fas were examined in OECs and the level of FasL was measured in CM. The gld mice were used to confirm a role for the Fas system in triggering apoptosis of embryos and oviducts. This study showed that blastocyst development was unaffected when mouse zygotes were cultured in CZB medium containing various concentrations of CRH/Cort but was impaired when embryos were cultured with CRH/Cort plus OECs or in CM conditioned with OECs pretreated with CRH/Cort (treatment CM). Culture in treatment-CM induced oxidative stress and apoptosis in embryos. Preimplantation embryos expressed GR and Fas at all stages and CRHR1 at the blastocyst stage only. Mouse 4-cell embryos and blastocysts expressed HSD2 but not HSD1. Culture of OECs with CRH/Cort increased their oxidative stress, apoptosis, CRHR1, Fas and FasL while decreasing their GR and growth factors. Blastocyst development in treatment-CM conditioned with OECs from gld mice harboring FasL mutations was superior to treatment-CM conditioned with wild-type mouse OECs. The results suggest that CRH/Cort impairs embryo development indirectly by inducing oviductal apoptosis via activating the Fas system. The insensitivity of preimplantation embryos to CRH and corticosterone is due to, respectively, a lack of CRHR and the exclusive expression of HSD2 that inactivate corticosterone. Not applicable. Although significant, the conclusions were drawn from limited results obtained using mice and thus they need further verification in other species. For example, bovine embryos express both HSD1 and HSD2 at all the preimplantation stages whereas mouse preimplantation embryos express HSD2 exclusively without HSD1. The data are important for our understanding of the mechanisms by which stress affects female reproduction in both human and animals, as early stages of pregnancy are considered more vulnerable to stress than the late stages. This study was supported by grants from the National Basic Research Program of China (Nos. 2014CB138503 and 2012CB944403), the China National Natural Science Foundation (Nos. 31272444 and 30972096) and the Animal breeding improvement program of Shandong Province. All authors declare that their participation in the study did not involve factual or potential conflicts of interests. © The Author 2017. Published by Oxford University Press on behalf of the European Society of Human Reproduction and Embryology. All rights reserved. For Permissions, please e-mail: journals.permissions@oup.com

  6. Caveolin-1 down-regulation is required for Wnt5a-Frizzled 2 signalling in Ha-RasV12 -induced cell transformation.

    PubMed

    Lin, Hsiu-Kuan; Lin, Hsi-Hui; Chiou, Yu-Wei; Wu, Ching-Lung; Chiu, Wen-Tai; Tang, Ming-Jer

    2018-05-01

    Caveolin-1 (Cav1) is down-regulated during MK4 (MDCK cells harbouring inducible Ha-Ras V12 gene) transformation by Ha-Ras V12 . Cav1 overexpression abrogates the Ha-Ras V12 -driven transformation of MK4 cells; however, the targeted down-regulation of Cav1 is not sufficient to mimic this transformation. Cav1-silenced cells, including MK4/shCav1 cells and MDCK/shCav1 cells, showed an increased cell area and discontinuous junction-related proteins staining. Cellular and mechanical transformations were completed when MDCK/shCav1 cells were treated with medium conditioned by MK4 cells treated with IPTG (MK4+I-CM) but not with medium conditioned by MK4 cells. Nanoparticle tracking analysis showed that Ha-Ras V12 -inducing MK4 cells increased exosome-like microvesicles release compared with their normal counterparts. The cellular and mechanical transformation activities of MK4+I-CM were abolished after heat treatment and exosome depletion and were copied by exosomes derived from MK4+I-CM (MK4+I-EXs). Wnt5a, a downstream product of Ha-Ras V12 , was markedly secreted by MK4+I-CM and MK4+I-EXs. Suppression of Wnt5a expression and secretion using the porcupine inhibitor C59 or Wnt5a siRNA inhibited the Ha-Ras V12 - and MK4+I-CM-induced transformation of MK4 cells and MDCK/shCav1 cells, respectively. Cav1 down-regulation, either by Ha-Ras V12 or targeted shRNA, increased frizzled-2 (Fzd2) protein levels without affecting its mRNA levels, suggesting a novel role of Cav1 in negatively regulating Fzd2 expression. Additionally, silencing Cav1 facilitated the internalization of MK4+I-EXs in MDCK cells. These data suggest that Cav1-dependent repression of Fzd2 and exosome uptake is potentially relevant to its antitransformation activity, which hinders the activation of Ha-Ras V12 -Wnt5a-Stat3 pathway. Altogether, these results suggest that both decreasing Cav1 and increasing exosomal Wnt5a must be implemented during Ha-Ras V12 -driven cell transformation. © 2018 The Authors. Journal of Cellular and Molecular Medicine published by John Wiley & Sons Ltd and Foundation for Cellular and Molecular Medicine.

  7. Absorption and emission of single attosecond light pulses in an autoionizing gaseous medium dressed by a time-delayed control field

    NASA Astrophysics Data System (ADS)

    Chu, Wei-Chun; Lin, C. D.

    2013-01-01

    An extreme ultraviolet (EUV) single attosecond pulse passing through a laser-dressed dense gas is studied theoretically. The weak EUV pulse pumps the helium gas from the ground state to the 2s2p(1P) autoionizing state, which is coupled to the 2s2(1S) autoionizing state by a femtosecond infrared laser with the intensity in the order of 1012 W/cm2. The simulation shows how the transient absorption and emission of the EUV are modified by the coupling laser. A simple analytical expression for the atomic response derived for δ-function pulses reveals the strong modification of the Fano lineshape in the spectra, where these features are quite universal and remain valid for realistic pulse conditions. We further account for the propagation of pulses in the medium and show that the EUV signal at the atomic resonance can be enhanced in the gaseous medium by more than 50% for specifically adjusted laser parameters, and that this enhancement persists as the EUV propagates in the gaseous medium. Our result demonstrates the high-level control of nonlinear optical effects that are achievable with attosecond pulses.

  8. Characteristics of Navy Medium-Weight High-Impact Shock Machine

    DTIC Science & Technology

    1951-09-14

    Ar 2-4~ -C 0 ofoL "- oili 0jit -1 b. V’ A . -- c- MC a y - w CmLUA ~ E~8.9. - flb. A 9 Er IL v II - W43 4P " It IIf ~NRL REPORT I CHARACTERISTICS OF...this machine under specification operation. A comnarison of datta is intended to correlate this shock to shipboard shock experienced in actual combat...table reversal are discussed, and it is shown that this secondsry shock can be the largest under certain condition. Theoretical equations of motion are

  9. Near-infrared absorption spectroscopy of interstellar hydrocarbon grains

    NASA Astrophysics Data System (ADS)

    Pendleton, Y. J.; Sandford, S. A.; Allamandola, L. J.; Tielens, A. G. G. M.; Sellgren, K.

    1994-12-01

    We present new 3600 - 2700/cm (2.8 - 3.7 micrometer) spectra of objects whose extinction is dominated by dust in the diffuse interstellar medium. The observations presented here augment an ongoing study of the organic component of the diffuse interstellar medium. These spectra contain a broad feature centered near 3300/cm (3.0 micrometers) and/or a feature with a more complex profile near 2950/cm (3.4 micrometers), the latter of which is attributed to saturated aliphatic hydrocarbons in interstellar grains and is the primary interest of this paper. As in our earlier work, the similarity of the absorption bands near 2950/cm (3.4 micrometers) along different lines of sight and the correlation of these features with interstellar extinction reveal that the carrier of this band lies in the dust in the diffuse interstellar medium (DISM). At least 2.5% of the cosmic carbon in the local interstellar medium and 4% toward the Galactic center is tied up in the carrier of the 2950/cm (3.4 micrometer) band. The spectral structure of the diffuse dust hydrocarbon C-H stretch absorption features is reasonably similar to UV photolyzed laboratory ice residues and is quite similar to the carbonaceous component of the Murchison meteorite. The similarity between the DISM and the meteoritic spectrum suggests that some of the interstellar material originally incorporated into the solar nebula may have survived relatively untouched in primitive solar system bodies. Comparisons of the DISM spectrum to hydrogenated amorphous carbon and quenched carbonaceous composite are also presented. The AV/tau ratio for the 2950/cm (3.4 micrometer) feature is lower toward the Galactic center than toward sources in the local solar neighborhood (approximately 150 for the Galactic center sources vs. approximately 250 for the local ISM sources). A similar trend has been observed previously for silicates in the diffuse medium by Roche & Aitken, suggesting that (1) the silicate and carbonaceous materials in the DISM may be physically correlated and (2) there is either dust compositional variation in the galaxy or galactic variation in the grain population density distribution. We also note a possible absorption feature near 3050/cm (3.28 micrometers), a wavelength position that is characteristic of polycyclic aromatic hydrocarbons (PAHs).

  10. Near-infrared absorption spectroscopy of interstellar hydrocarbon grains

    NASA Technical Reports Server (NTRS)

    Pendleton, Y. J.; Sandford, S. A.; Allamandola, L. J.; Tielens, A. G. G. M.; Sellgren, K.

    1994-01-01

    We present new 3600 - 2700/cm (2.8 - 3.7 micrometer) spectra of objects whose extinction is dominated by dust in the diffuse interstellar medium. The observations presented here augment an ongoing study of the organic component of the diffuse interstellar medium. These spectra contain a broad feature centered near 3300/cm (3.0 micrometers) and/or a feature with a more complex profile near 2950/cm (3.4 micrometers), the latter of which is attributed to saturated aliphatic hydrocarbons in interstellar grains and is the primary interest of this paper. As in our earlier work, the similarity of the absorption bands near 2950/cm (3.4 micrometers) along different lines of sight and the correlation of these features with interstellar extinction reveal that the carrier of this band lies in the dust in the diffuse interstellar medium (DISM). At least 2.5% of the cosmic carbon in the local interstellar medium and 4% toward the Galactic center is tied up in the carrier of the 2950/cm (3.4 micrometer) band. The spectral structure of the diffuse dust hydrocarbon C-H stretch absorption features is reasonably similar to UV photolyzed laboratory ice residues and is quite similar to the carbonaceous component of the Murchison meteorite. The similarity between the DISM and the meteoritic spectrum suggests that some of the interstellar material originally incorporated into the solar nebula may have survived relatively untouched in primitive solar system bodies. Comparisons of the DISM spectrum to hydrogenated amorphous carbon and quenched carbonaceous composite are also presented. The A(sub V)/tau ratio for the 2950/cm (3.4 micrometer) feature is lower toward the Galactic center than toward sources in the local solar neighborhood (approximately 150 for the Galactic center sources vs. approximately 250 for the local ISM sources). A similar trend has been observed previously for silicates in the diffuse medium by Roche & Aitken, suggesting that (1) the silicate and carbonaceous materials in the DISM may be physically correlated and (2) there is either dust compositional variation in the galaxy or galactic variation in the grain population density distribution. We also note a possible absorption feature near 3050/cm (3.28 micrometers), a wavelength position that is characteristic of polycyclic aromatic hydrocarbons (PAHs).

  11. Inbuilt Potential of YEM Medium and Its Constituents to Generate Ag/Ag2O Nanoparticles

    PubMed Central

    Yamal, G.; Sharmila, P.; Rao, K. S.; Pardha-Saradhi, P.

    2013-01-01

    We discovered that Yeast Extract Mannitol (YEM) medium possessed immense potential to generate silver nanoparticles from AgNO3 upon autoclaving, which was evident from (i) alteration in color of the medium; (ii) peak at ∼410 nm in UV-Vis spectrum due to surface plasmon resonance specific to silver nanoparticles; and (iii) TEM investigations. TEM coupled with EDX confirmed that distinct nanoparticles were composed of silver. Yeast extract and mannitol were key components of YEM medium responsible for the formation of nanoparticles. PXRD analysis indicated crystalline geometry and Ag/Ag2O phases in nanoparticles generated with YEM medium, yeast extract and mannitol. Our investigations also revealed that both mannitol and yeast extract possessed potential to convert ∼80% of silver ions in 0.5 mM AgNO3 to nanoparticles, on autoclaving for 30 min at 121°C under a pressure of 1.06 kg/cm2. Addition of filter sterilized AgNO3 under ambient conditions to pre-autoclaved YEM medium and yeast extract brought about color change due to the formation of silver nanoparticles, but required prolonged duration. In general, even after 72 h intensity of color was significantly less than that recorded following autoclaving. Silver nanoparticles formed at room temperature were more heterogeneous compared to that obtained upon autoclaving. In summary, our findings demonstrated that (i) YEM medium and its constituents promote synthesis of silver nanoparticles; and (ii) autoclaving enhances rapid synthesis of silver nanoparticles by YEM medium, yeast extract and mannitol. PMID:23626722

  12. Porous structures of polymer films prepared by spin coating with mixed solvents under humid condition.

    PubMed

    Park, Min Soo; Joo, Wonchul; Kim, Jin Kon

    2006-05-09

    We investigate the effects of interfacial energy between water and solvent as well as polymer concentration on the formation of porous structures of polymer films prepared by spin coating of cellulose acetate butyrate (CAB) in mixed solvent of tetrahydrofuran (THF) and chloroform under humid condition. The interfacial energy between water and the solvent was gradually changed by the addition of chloroform to the solvent. At a high polymer concentration (0.15 g/cm3 in THF), porous structures were limited only at the top surfaces of CAB films, regardless of interfacial energies, due to the high viscosity of the solution. At a medium concentration (approximately 0.08 g/cm3 in THF), CAB film had relatively uniform pores at the top surface and very small pores inside the film because of the mixing of the water droplets with THF solution. When chloroform was added to THF, pores at the inner CAB film had a comparable size with those at the top surface because of the reduced degree of the mixing between the water droplets and the mixed solvent. A further decrease in polymer concentration (0.05 g/cm3 in THF) caused the final films to have a two-layer porous structure, and the size of pores at each layer was almost the same.

  13. 78 FR 26709 - Atlantic Highly Migratory Species; Atlantic Bluefin Tuna Fisheries

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-05-08

    ... the Atlantic bluefin tuna (BFT) General category daily retention limit to three large medium or giant... medium or giant BFT (measuring 73 inches (185 cm) curved fork length (CFL) or greater) per vessel per day... 2012 fishing year, NMFS adjusted the General category limit from the default level of one large medium...

  14. Infrared spectroscopic examination of the interaction of urea with the naturally occurring zeolite clinoptilolite

    USGS Publications Warehouse

    Byler, D.M.; Gerasimowicz, W.V.; Stockette, V.M.; Eberl, D.D.

    1991-01-01

    Infrared spectroscopy has shown for the first time that the naturally occurring zeolite clinoptilolite can absorb urea, (NH2)2CO, under ambient conditions from either aqueous or ethanolic solutions. The two strongest NH stretching bands at 3441 and 3344 cm-1 in pure, solid urea shift to higher frequency (about 3504 and 3401 cm-1) after absorption. Two of the four urea bands in the 1800-1300 cm-1 range (at 1683 and 1467 cm-1) undergo marked downward shifts to about 1670 and 1445 cm-1. The other two bands show little change in frequency. The strong band at 1602 cm-1, however, diminishes in intensity to little more than an ill-defined shoulder on the 1626-cm-1 peak. When clinoptilolite is heated to 450 ??C and then treated with molten urea (ca. 140 ??C) for several minutes, and finally washed twice with ethanol to remove excess unreacted urea, further changes become apparent in the spectrum of the urea-treated clinoptilolite. The two NH stretching bands broaden without significant change in frequency. Two new bands appear in the midfrequency range at 1777 (weak) and 1719 (medium strong) cm-1. Of the four original midfrequency peaks, the one at 1602 cm-1 is now absent. Two others (1627 and 1440 cm-1) exhibit little change, while the fourth has broadened and shifted down to 1663 cm-1, where it appears as a shoulder on the band at 1627 cm-1. Both treatments clearly induce interaction between urea and the zeolite which seems to result in significant modifications in the nature of the hydrogen bonding of the substrate. ?? 1991.

  15. Extremely low-loss, dispersion flattened porous-core photonic crystal fiber for terahertz regime

    NASA Astrophysics Data System (ADS)

    Islam, Saiful; Islam, Mohammad Rakibul; Faisal, Mohammad; Arefin, Abu Sayeed Muhammad Shamsul; Rahman, Hasan; Sultana, Jakeya; Rana, Sohel

    2016-07-01

    A porous-core octagonal photonic crystal fiber (PC-OPCF) with ultralow effective material loss (EML), high core power fraction, and ultra flattened dispersion is proposed for terahertz (THz) wave propagation. At an operating frequency of 1 THz and core diameter of 345 μm, simulation results display an extremely low EML of 0.047 cm-1, 49.1% power transmission through core air holes, decreased confinement loss with the increase of frequency, and dispersion variation of 0.15 ps/THz/cm. In addition, the proposed PCF can successfully operate in single-mode condition. All the simulations are performed with finite-element modeling package, COMSOL v4.2. The design can be fabricated using a stacking and drilling method. Thus, the proposed fiber has the potential of being an effective transmission medium of broadband THz waves.

  16. Human Umbilical Cord Wharton's Jelly Stem Cell Conditioned Medium Induces Tumoricidal Effects on Lymphoma Cells Through Hydrogen Peroxide Mediation.

    PubMed

    Lin, Hao Daniel; Fong, Chui-Yee; Biswas, Arijit; Choolani, Mahesh; Bongso, Ariff

    2016-09-01

    Several groups have reported that human umbilical cord Wharton's jelly stem cells (hWJSCs) possess unique tumoricidal properties against many cancers. However, the exact mechanisms as to how hWJSCs inhibit tumor growth are not known. Recent evidence suggests that exposure of cancer cells to high hydrogen peroxide (H2 O2 ) levels from H2 O2 -releasing drugs causes their death. We therefore explored whether the tumoricidal effect of hWJSCs on lymphoma cells was mediated via H2 O2 . We first exposed lymphoma cells to six different molecular weight cut-off (MWCO) concentrates of hWJSC-conditioned medium (hWJSC-CM) (3, 5, 10, 30, 50, 100 kDa) for 48 h. Since, the 3 kDa-MWCO concentrate showed the greatest cell inhibition we then investigated whether the tumoricidal effect of the specific 3 kDa-MWCO concentrate on two different lymphoma cell lines (Ramos and Toledo) was mediated via accumulation of H2 O2 . We used a battery of assays (MTT, propidium iodide, mitochondria membrane potential, apoptosis, cell cycle, oxidative stress enzymes, hydrogen peroxide, mitochondrial superoxide, hydroxyl radical, peroxynitrile anion, and lipid peroxidation) to test this mechanism. The hWJSC-CM-3 kDa MWCO concentrate significantly decreased cell viability and mitochondrial membrane potential and increased cell death and apoptosis in both lymphoma cell lines. There were significant increases in superoxide dismutase with concomitant decreases in glutathione peroxidase, catalase, and thioredoxin peroxidase activities. H2 O2 levels, mitochondrial superoxide, hydroxyl radical, peroxynitrile anion, and lipid peroxidation were also significantly increased in both lymphoma cell lines. The results suggested that the hWJSC-CM-3 kDa MWCO concentrate regulates cellular H2 O2 leading to a tumoricidal effect and may thus be a promising anti-lymphoma agent. J. Cell. Biochem. 117: 2045-2055, 2016. © 2016 Wiley Periodicals, Inc. © 2016 Wiley Periodicals, Inc.

  17. Nutrient supplemented serum-free medium increases cardiomyogenesis efficiency of human pluripotent stem cells.

    PubMed

    Ting, Sherwin; Lecina, Marti; Chan, Yau-Chi; Tse, Hung Fat; Reuveny, Shaul; Oh, Steve Kw

    2013-07-26

    To development of an improved p38 MAPK inhibitor-based serum-free medium for embryoid body cardiomyocyte differentiation of human pluripotent stem cells. Human embryonic stem cells (hESC) differentiated to cardiomyocytes (CM) using a p38 MAPK inhibitor (SB203580) based serum-free medium (SB media). Nutrient supplements known to increase cell viability were added to SB medium. The ability of these supplements to improve cardiomyogenesis was evaluated by measurements of cell viability, total cell count, and the expression of cardiac markers via flow cytometry. An improved medium containing Soy hydrolysate (HySoy) and bovine serum albumin (BSA) (SupSB media) was developed and tested on 2 additional cell lines (H1 and Siu-hiPSC). Characterization of the cardiomyocytes was done by immunohistochemistry, electrophysiology and quantitative real-time reverse transcription-polymerase chain reaction. hESC cell line, HES-3, differentiating in SB medium for 16 d resulted in a cardiomyocyte yield of 0.07 ± 0.03 CM/hESC. A new medium (SupSB media) was developed with the addition of HySoy and BSA to SB medium. This medium resulted in 2.6 fold increase in cardiomyocyte yield (0.21 ± 0.08 CM/hESC). The robustness of SupSB medium was further demonstrated using two additional pluripotent cell lines (H1, hESC and Siu1, hiPSC), showing a 15 and 9 fold increase in cardiomyocyte yield respectively. The age (passage number) of the pluripotent cells did not affect the cardiomyocyte yields. Embryoid body (EB) cardiomyocytes formed in SupSB medium expressed canonical cardiac markers (sarcomeric α-actinin, myosin heavy chain and troponin-T) and demonstrated all three major phenotypes: nodal-, atrial- and ventricular-like. Electrophysiological characteristics (maximum diastolic potentials and action potential durations) of cardiomyocytes derived from SB and SupSB media were similar. The nutrient supplementation (HySoy and BSA) leads to increase in cell viability, cell yield and cardiac marker expression during cardiomyocyte differentiation, translating to an overall increase in cardiomyocyte yield.

  18. Percutaneous Radiofrequency Ablation with Multiple Electrodes for Medium-Sized Hepatocellular Carcinomas

    PubMed Central

    Lee, Jung; Yoon, Jung-Hwan; Lee, Jae Young; Kim, Se Hyung; Lee, Jeong Eun; Han, Joon Koo; Choi, Byung Ihn

    2012-01-01

    Objective To prospectively evaluate the safety and short-term therapeutic efficacy of switching monopolar radiofrequency ablation (RFA) with multiple electrodes to treat medium-sized (3.1-5.0 cm), hepatocellular carcinomas (HCC). Materials and Methods In this prospective study, 30 patients with single medium-sized HCCs (mean, 3.5 cm; range, 3.1-4.4 cm) were enrolled. The patients were treated under ultrasonographic guidance by percutaneous switching monopolar RFA with a multichannel RF generator and two or three internally cooled electrodes. Contrast-enhanced CT scans were obtained immediately after RFA, and the diameters and volume of the ablation zones were then measured. Follow-up CT scans were performed at the first month after ablation and every three months thereafter. Technical effectiveness, local progression and remote recurrence of HCCs were determined. Results There were no major immediate or periprocedural complications. However, there was one bile duct stricture during the follow-up period. Technical effectiveness was achieved in 29 of 30 patients (97%). The total ablation time of the procedures was 25.4 ± 8.9 minutes. The mean ablation volume was 73.8 ± 56.4 cm3 and the minimum diameter was 4.1 ± 7.3 cm. During the follow-up period (mean, 12.5 months), local tumor progression occurred in three of 29 patients (10%) with technical effectiveness, while new HCCs were detected in six of 29 patients (21%). Conclusion Switching monopolar RFA with multiple electrodes in order to achieve a sufficient ablation volume is safe and efficient. This method also showed relatively successful therapeutic effectiveness on short-term follow up for the treatment of medium-sized HCCs. PMID:22247634

  19. Peripheral Nerve Regeneration by Secretomes of Stem Cells from Human Exfoliated Deciduous Teeth.

    PubMed

    Sugimura-Wakayama, Yukiko; Katagiri, Wataru; Osugi, Masashi; Kawai, Takamasa; Ogata, Kenichi; Sakaguchi, Kohei; Hibi, Hideharu

    2015-11-15

    Peripheral nerve regeneration across nerve gaps is often suboptimal, with poor functional recovery. Stem cell transplantation-based regenerative therapy is a promising approach for axon regeneration and functional recovery of peripheral nerve injury; however, the mechanisms remain controversial and unclear. Recent studies suggest that transplanted stem cells promote tissue regeneration through a paracrine mechanism. We investigated the effects of conditioned media derived from stem cells from human exfoliated deciduous teeth (SHED-CM) on peripheral nerve regeneration. In vitro, SHED-CM-treated Schwann cells exhibited significantly increased proliferation, migration, and the expression of neuron-, extracellular matrix (ECM)-, and angiogenesis-related genes. SHED-CM stimulated neuritogenesis of dorsal root ganglia and increased cell viability. Similarly, SHED-CM enhanced tube formation in an angiogenesis assay. In vivo, a 10-mm rat sciatic nerve gap model was bridged by silicon conduits containing SHED-CM or serum-free Dulbecco's modified Eagle's medium. Light and electron microscopy confirmed that the number of myelinated axons and axon-to-fiber ratio (G-ratio) were significantly higher in the SHED-CM group at 12 weeks after nerve transection surgery. The sciatic functional index (SFI) and gastrocnemius (target muscle) wet weight ratio demonstrated functional recovery. Increased compound muscle action potentials and increased SFI in the SHED-CM group suggested sciatic nerve reinnervation of the target muscle and improved functional recovery. We also observed reduced muscle atrophy in the SHED-CM group. Thus, SHEDs may secrete various trophic factors that enhance peripheral nerve regeneration through multiple mechanisms. SHED-CM may therefore provide a novel therapy that creates a more desirable extracellular microenvironment for peripheral nerve regeneration.

  20. Potential therapeutic effect of the secretome from human uterine cervical stem cells against both cancer and stromal cells compared with adipose tissue stem cells.

    PubMed

    Eiró, Noemí; Sendon-Lago, Juan; Seoane, Samuel; Bermúdez, María A; Lamelas, Maria Luz; Garcia-Caballero, Tomás; Schneider, José; Perez-Fernandez, Roman; Vizoso, Francisco J

    2014-11-15

    Evidences indicate that tumor development and progression towards a malignant phenotype depend not only on cancer cells themselves, but are also deeply influenced by tumor stroma reactivity. The present study uses mesenchymal stem cells from normal human uterine cervix (hUCESCs), isolated by the minimally invasive method of routine Pap cervical smear, to study their effect on the three main cell types in a tumor: cancer cells, fibroblasts and macrophages. Administration of hUCESCs-conditioned medium (CM) to a highly invasive breast cancer MDA-MB-231 cell line and to human breast tumors with high cell proliferation rates had the effect of reducing cell proliferation, modifying the cell cycle, inducing apoptosis, and decreasing invasion. In a xenograft mouse tumor model, hUCESCs-CM reduced tumor growth and increased overall survival. In cancer-associated fibroblasts, administration of hUCESCs-CM resulted in reduced cell proliferation, greater apoptosis and decreased invasion. In addition, hUCESCs-CM inhibited and reverted macrophage differentiation. The analysis of hUCESCs-CM (fresh and lyophilized) suggests that a complex paracrine signaling network could be implicated in the anti-tumor potential of hUCESCs. In light of their anti-tumor potential, the easy cell isolation method, and the fact that lyophilization of their CM conserves original properties make hUCESCs good candidates for experimental or clinical applications in anticancer therapy.

  1. Influence of humidity on the initial emittable concentration of formaldehyde and hexaldehyde in building materials: experimental observation and correlation.

    PubMed

    Huang, Shaodan; Xiong, Jianyin; Cai, Chaorui; Xu, Wei; Zhang, Yinping

    2016-03-30

    Humidity is one of the main environmental factors affecting the emission rate and key parameters of formaldehyde and volatile organic compounds (VOCs) from building materials. Meanwhile, the initial emittable concentration (Cm,0) is proved to be the most sensitive key parameter to the emission behaviours. However, there is no report on the relationship between humidity and Cm,0. In this paper, Cm,0 of formaldehyde and hexaldehyde from a type of medium density fiberboard in absolute humidity (AH) range of 4.6-19.6 g/m(3) at 25 °C were tested by virtue of a C-history method. Experimental results indicate that Cm,0 is dramatically dependent on AH, increased by 10 and 2 times for formaldehyde and hexaldehyde when AH rising from 4.6 g/m(3) to 19.6 g/m(3). A linear relationship between the logarithm of Cm,0 and AH is obtained based on the measured results. In addition, a correlation characterizing the association of emission rate and AH is derived. The effectiveness of the correlation is verified with our experimental results as well as data from literature. With the correlations, the Cm,0 or emission rate different from the test AH conditions can be conveniently obtained. This study should be useful for predicting the emission characteristics of humidity changing scenarios and for source control.

  2. Structural differences between cell matrix and culture medium N-linked oligosaccharides on heavy proteochondroitin sulfate proteoglycan (PCS-H)

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

    Cioffi, L.; Conrad, H.E.

    1986-05-01

    Tibial chondrocytes were labeled metabolically with /sup 3/H-man and the PCS-H was isolated from the culture medium (CM) and the cell matrix (Ma) pools. Equal amounts of /sup 3/H were incorporated into the PCS-H of the CM and Ma pools. The PCS-H pools were digested with thermolysin, Chondroitinase, and then N-glycanase, and the N-linked oligosaccharides were chromatographed on Con-A Sepharose. The ratios of complex to high mannose oligosaccharides for the CM and Ma were 6.1 and 2.6, respectively. More than 60% of the complex CM N-linked oligosaccharides were charged species whereas only 40% of the Ma N-linked oligosaccharides were charged.more » The oligosaccharides were analyzed by HPLC. Both complex and high mannose oligosaccharides found in the PCS-H of the CM and Ma pools were mixtures of identical structures but the amounts of each structure in the two pools showed marked differences. These observations indicate that distinct PCS-H species are found in the CM and Ma pools.« less

  3. Metabolomic comparison between wild Ophiocordyceps sinensis and artificial cultured Cordyceps militaris.

    PubMed

    Chen, Lin; Liu, Yuetao; Guo, Qingfeng; Zheng, Qingxia; Zhang, Wancun

    2018-05-11

    A systematic study on the metabolome differences between wild Ophiocordyceps sinensis and artificial cultured Cordyceps militaris was conducted using liquid chromatography-mass spectrometry. Principal component analysis and orthogonal projection on latent structure-discriminant analysis results showed that C. militaris grown on solid rice medium (R-CM) and C. militaris grown on tussah pupa (T-CM) evidently separated and individually separated from wild O. sinensis, indicating metabolome difference among wild O. sinensis, R-CM and T-CM. The metabolome differences between R-CM and T-CM indicated that C. militaris could accommodate to culture medium by differential metabolic regulation. Hierarchical clustering analysis was further performed to cluster the differential metabolites and samples based on their metabolic similarity. The higher content of amino acids (pyroglutamic acid, glutamic acid, histidine, phenylalanine and arginine), unsaturated fatty acid (linolenic acid and linoleic acid), peptides, mannitol, adenosine and succinoadenosine in O. sinensis make it as an excellent choice as a traditional Chinese medicine for invigoration or nutritional supplementation. Similar compositions with O. sinensis and easy cultivation make artificially cultured C. militaris a possible alternative to O. sinensis. Copyright © 2018 John Wiley & Sons, Ltd.

  4. Influence of Silica Nanoparticles on Antioxidant Potential of Bacillus subtilis IMV B-7023.

    PubMed

    Skorochod, Iryna O; Roy, Alla O; Kurdish, Ivan K

    2016-12-01

    It was found that if introduced into a nutrient medium of 0.05-1 g/L nano-SiO2, the oxidant activity (OA) of the culture medium (CM) of bacilli increased by 43.2-60.1 % and the antioxidant activity (AA) decreased by 4.5-11.8 %. SiO2 nanoparticles had different effects on antiradical activity (ARA) of the CM of Bacillus subtilis IMV B-7023. In particular, nano-SiO2 had no significant effect on the ability of the CM of bacilli to inactivate the 2.2-diphenyl-1-picrylhydrazyl (DPPH·) free radical. However, for the content of the nanomaterial of 0.01-1 g/L decreased hydroxyl radical scavenging in the CM of B. subtilis IMV B-7023 on 7.2-17.6 % compared with a control. Low doses of silica nanoparticles stimulated the reducing power of the CM of bacteria and then highly suppressed it.

  5. 76 FR 52886 - Atlantic Highly Migratory Species; Atlantic Bluefin Tuna Fisheries

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-08-24

    ... be adjusted from one to three large medium or giant BFT for the September, October- November, and... medium or giant BFT (measuring 73 inches (185 cm) curved fork length or greater) per vessel per day/trip... increase or decrease the daily retention limit of large medium and giant BFT over a range of zero to a...

  6. Mesenchymal stem cells and their conditioned medium improve integration of purified induced pluripotent stem cell-derived cardiomyocyte clusters into myocardial tissue.

    PubMed

    Rubach, Martin; Adelmann, Roland; Haustein, Moritz; Drey, Florian; Pfannkuche, Kurt; Xiao, Bing; Koester, Annette; Udink ten Cate, Floris E A; Choi, Yeong-Hoon; Neef, Klaus; Fatima, Azra; Hannes, Tobias; Pillekamp, Frank; Hescheler, Juergen; Šarić, Tomo; Brockmeier, Konrad; Khalil, Markus

    2014-03-15

    Induced pluripotent stem cell-derived cardiomyocytes (iPS-CMs) might become therapeutically relevant to regenerate myocardial damage. Purified iPS-CMs exhibit poor functional integration into myocardial tissue. The aim of this study was to investigate whether murine mesenchymal stem cells (MSCs) or their conditioned medium (MScond) improves the integration of murine iPS-CMs into myocardial tissue. Vital or nonvital embryonic murine ventricular tissue slices were cocultured with purified clusters of iPS-CMs in combination with murine embryonic fibroblasts (MEFs), MSCs, or MScond. Morphological integration was assessed by visual scoring and functional integration by isometric force and field potential measurements. We observed a moderate morphological integration of iPS-CM clusters into vital, but a poor integration into nonvital, slices. MEFs and MSCs but not MScond improved morphological integration of CMs into nonvital slices and enabled purified iPS-CMs to confer force. Coculture of vital slices with iPS-CMs and MEFs or MSCs resulted in an improved electrical integration. A comparable improvement of electrical coupling was achieved with the cell-free MScond, indicating that soluble factors secreted by MSCs were involved in electrical coupling. We conclude that cells such as MSCs support the engraftment and adhesion of CMs, and confer force to noncontractile tissue. Furthermore, soluble factors secreted by MSCs mediate electrical coupling of purified iPS-CM clusters to myocardial tissue. These data suggest that MSCs may increase the functional engraftment and therapeutic efficacy of transplanted iPS-CMs into infarcted myocardium.

  7. Mesenchymal Stem Cells and Their Conditioned Medium Improve Integration of Purified Induced Pluripotent Stem Cell–Derived Cardiomyocyte Clusters into Myocardial Tissue

    PubMed Central

    Rubach, Martin; Adelmann, Roland; Haustein, Moritz; Drey, Florian; Pfannkuche, Kurt; Xiao, Bing; Koester, Annette; Udink ten Cate, Floris E.A.; Choi, Yeong-Hoon; Neef, Klaus; Fatima, Azra; Hannes, Tobias; Pillekamp, Frank; Hescheler, Juergen; Šarić, Tomo; Brockmeier, Konrad

    2014-01-01

    Induced pluripotent stem cell–derived cardiomyocytes (iPS-CMs) might become therapeutically relevant to regenerate myocardial damage. Purified iPS-CMs exhibit poor functional integration into myocardial tissue. The aim of this study was to investigate whether murine mesenchymal stem cells (MSCs) or their conditioned medium (MScond) improves the integration of murine iPS-CMs into myocardial tissue. Vital or nonvital embryonic murine ventricular tissue slices were cocultured with purified clusters of iPS-CMs in combination with murine embryonic fibroblasts (MEFs), MSCs, or MScond. Morphological integration was assessed by visual scoring and functional integration by isometric force and field potential measurements. We observed a moderate morphological integration of iPS-CM clusters into vital, but a poor integration into nonvital, slices. MEFs and MSCs but not MScond improved morphological integration of CMs into nonvital slices and enabled purified iPS-CMs to confer force. Coculture of vital slices with iPS-CMs and MEFs or MSCs resulted in an improved electrical integration. A comparable improvement of electrical coupling was achieved with the cell-free MScond, indicating that soluble factors secreted by MSCs were involved in electrical coupling. We conclude that cells such as MSCs support the engraftment and adhesion of CMs, and confer force to noncontractile tissue. Furthermore, soluble factors secreted by MSCs mediate electrical coupling of purified iPS-CM clusters to myocardial tissue. These data suggest that MSCs may increase the functional engraftment and therapeutic efficacy of transplanted iPS-CMs into infarcted myocardium. PMID:24219308

  8. Snowmobile impacts on snowpack physical and mechanical properties

    NASA Astrophysics Data System (ADS)

    Fassnacht, Steven R.; Heath, Jared T.; Venable, Niah B. H.; Elder, Kelly J.

    2018-03-01

    Snowmobile use is a popular form of winter recreation in Colorado, particularly on public lands. To examine the effects of differing levels of use on snowpack properties, experiments were performed at two different areas, Rabbit Ears Pass near Steamboat Springs and at Fraser Experimental Forest near Fraser, Colorado USA. Differences between no use and varying degrees of snowmobile use (low, medium and high) on shallow (the operational standard of 30 cm) and deeper snowpacks (120 cm) were quantified and statistically assessed using measurements of snow density, temperature, stratigraphy, hardness, and ram resistance from snow pit profiles. A simple model was explored that estimated snow density changes from snowmobile use based on experimental results. Snowpack property changes were more pronounced for thinner snow accumulations. When snowmobile use started in deeper snow conditions, there was less difference in density, hardness, and ram resistance compared to the control case of no snowmobile use. These results have implications for the management of snowmobile use in times and places of shallower snow conditions where underlying natural resources could be affected by denser and harder snowpacks.

  9. Application of potassium permanganate to spectrophotometric assay of metoclopramide hydrochloride in pharmaceuticals

    NASA Astrophysics Data System (ADS)

    Devi, O. Zenita; Basavaiah, K.; Vinay, K. B.

    2012-01-01

    Two simple, sensitive, and cost-effective spectrophotometric methods are described for the determination of metoclopramide hydrochloride (MCP) in pharmaceutical dosage forms. The methods are based on a redox reaction between MCP and KMnO4 in alkaline and acid media. Direct spectrophotometry (method A) involves treating MCP with permanganate in an NaOH medium and measuring a bluish green product at 610 nm. In indirect spectrophotometry (method B), MCP is treated with a fixed concentration of KMnO4 in an H2SO4 medium, and after a specified time, the unreacted KMnO4 is measured at 545 nm. Under optimum assay conditions, Beer's law is obeyed over the ranges of 0.75-12.0 and 2.5-30.0 g/ml for methods A and B, respectively. Molar absorptivity values are calculated to be 2.33•104 and 2.66•104 l/mol cm for methods A and B, respectively, and corresponding Sandell's sensitivity values are 0.015 and 0.013 g/cm2. Limits of detection (LOD) and quantification (LOQ) are also reported. The applicability of the developed methods was demonstrated by the determination of MCP in tablet and injection forms. The accuracy and reliability of the proposed methods were further ascertained by recovery studies via standard addition technique.

  10. Xanthan production on polyurethane foam and its enhancement by air pressure pulsation.

    PubMed

    Zhang, Zhi-guo; Chen, Hong-zhang

    2010-12-01

    In this study, we evaluated the feasibility of solid-state fermentation (SSF) on polyurethane foam (PUF) for xanthan production. The effects of air pressure pulsation (APP) on biomass accumulation and final xanthan concentration were also studied. Under suitable conditions (15% inoculum, 0.5-cm (side length) PUF cubes, 15 mL medium per gram cubes and 4.5 cm bed depth), the broth was dispersed on the PUF as a film. When the initial glucose concentration in the media was low (20 and 40 g L⁻¹), there was no significant difference between the final xanthan concentration in static SSF and submerged fermentation (SMF). When high initial glucose concentrations (60 and 80 g L⁻¹) were used, the final gum concentrations in SSF were much higher than those in SMF. When the APP technique was applied in xanthan production with a medium containing a high glucose concentration (80 g L⁻¹), the oxygen consumption rate of Xanthomonas campestris was significantly enhanced at the later stages of fermentation, and both the biomass and xanthan concentration were improved. The results indicated that SSF on PUF is suitable for xanthan preparation, especially when the initial glucose concentration ranged from 60 to 80 g L⁻¹. Those results also demonstrated that APP technology can be used to enhance xanthan yields.

  11. Factors secreted from dental pulp stem cells show multifaceted benefits for treating experimental rheumatoid arthritis.

    PubMed

    Ishikawa, Jun; Takahashi, Nobunori; Matsumoto, Takuya; Yoshioka, Yutaka; Yamamoto, Noriyuki; Nishikawa, Masaya; Hibi, Hideharu; Ishigro, Naoki; Ueda, Minoru; Furukawa, Koichi; Yamamoto, Akihito

    2016-02-01

    Rheumatoid arthritis (RA) is an autoimmune disease characterized by synovial hyperplasia and chronic inflammation, which lead to the progressive destruction of cartilage and bone in the joints. Numerous studies have reported that administrations of various types of MSCs improve arthritis symptoms in animal models, by paracrine mechanisms. However, the therapeutic effects of the secreted factors alone, without the cell graft, have been uncertain. Here, we show that a single intravenous administration of serum-free conditioned medium (CM) from human deciduous dental pulp stem cells (SHED-CM) into anti-collagen type II antibody-induced arthritis (CAIA), a mouse model of rheumatoid arthritis (RA), markedly improved the arthritis symptoms and joint destruction. The therapeutic efficacy of SHED-CM was associated with an induction of anti-inflammatory M2 macrophages in the CAIA joints and the abrogation of RANKL expression. SHED-CM specifically depleted of an M2 macrophage inducer, the secreted ectodomain of sialic acid-binding Ig-like lectin-9 (ED-Siglec-9), exhibited a reduced ability to induce M2-related gene expression and attenuate CAIA. SHED-CM also inhibited the RANKL-induced osteoclastogenesis in vitro. Collectively, our findings suggest that SHED-CM provides multifaceted therapeutic effects for treating CAIA, including the ED-Siglec-9-dependent induction of M2 macrophage polarization and inhibition of osteoclastogenesis. Thus, SHED-CM may represent a novel anti-inflammatory and reparative therapy for RA. Copyright © 2015 Elsevier Inc. All rights reserved.

  12. Predicting Transport of 3,5,6-Trichloro-2-Pyridinol Into Saliva Using a Combination Experimental and Computational Approach

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

    Smith, Jordan Ned; Carver, Zana A.; Weber, Thomas J.

    A combination experimental and computational approach was developed to predict chemical transport into saliva. A serous-acinar chemical transport assay was established to measure chemical transport with non-physiological (standard cell culture medium) and physiological (using surrogate plasma and saliva medium) conditions using 3,5,6-trichloro-2-pyridinol (TCPy) a metabolite of the pesticide chlorpyrifos. High levels of TCPy protein binding was observed in cell culture medium and rat plasma resulting in different TCPy transport behaviors in the two experimental conditions. In the non-physiological transport experiment, TCPy reached equilibrium at equivalent concentrations in apical and basolateral chambers. At higher TCPy doses, increased unbound TCPy was observed,more » and TCPy concentrations in apical and basolateral chambers reached equilibrium faster than lower doses, suggesting only unbound TCPy is able to cross the cellular monolayer. In the physiological experiment, TCPy transport was slower than non-physiological conditions, and equilibrium was achieved at different concentrations in apical and basolateral chambers at a comparable ratio (0.034) to what was previously measured in rats dosed with TCPy (saliva:blood ratio: 0.049). A cellular transport computational model was developed based on TCPy protein binding kinetics and accurately simulated all transport experiments using different permeability coefficients for the two experimental conditions (1.4 vs 0.4 cm/hr for non-physiological and physiological experiments, respectively). The computational model was integrated into a physiologically based pharmacokinetic (PBPK) model and accurately predicted TCPy concentrations in saliva of rats dosed with TCPy. Overall, this study demonstrates an approach to predict chemical transport in saliva potentially increasing the utility of salivary biomonitoring in the future.« less

  13. [Stability in association of the peripheral material with mitotic chromosomes].

    PubMed

    Kosykh, M I; Chentsov, Iu S

    2002-01-01

    The localization of nucleolar proteins (fibrillarin and B-23), and of the protein of interphase nuclear matrix (NMP-65) was studied in the perichromosomal material (CM) after of short hypotonic treatment (15% solution of Henks medium) on cultured pig embryonic kidney cells, followed by restoration of isotonic conditions. It is shown that during hypotonic shock the mitotic chromosomes demonstrate reversible swelling, but their periphery is bounded with a rim of PCM, containing antibodies to fibrillarin and NMP-65, but not to B-23. After returning the cells to the initial isotonic medium, all the three proteins can be detected again on the periphery of chromosomes. It suggests the existence of different stability in the association of free proteins with chromosome bodies. Besides, B-23 and fibrillarin could be visualized in residual nucleoli after a complete extraction of histones and DNA from nuclei.

  14. Raman Spectra of Nitrogen, Carbon Dioxide, and Hydrogen in a Methane Environment

    NASA Astrophysics Data System (ADS)

    Petrov, D. V.; Matrosov, I. I.; Sedinkin, D. O.; Zaripov, A. R.

    2018-01-01

    Changes in the Raman spectra of N2, H2, and CO2 are studied in the range of 200-3800 cm-1 depending on the concentration of surrounding CH4 molecules at a fixed medium pressure of 25 atm and temperature of 300 K. It has been found that changes in the spectral characteristics of purely rotational H2 lines in a CH4 medium are negligible, while the Q-branches of the v 1/2 v 2 Fermi dyad in CO2 become narrower and wavenumbers of its high-frequency component and v 1 band of N2 decrease. In addition, under these conditions, the ratio of intensities of the CO2 Fermi dyad Q-branch varies in proportion to the concentration of surrounding molecules of CH4. The obtained data will be used in diagnosing the composition of natural gas using Raman spectroscopy.

  15. Permeability of lipid bilayers to amino acids and phosphate

    NASA Technical Reports Server (NTRS)

    Chakrabarti, A. C.; Deamer, D. W.

    1992-01-01

    Permeability coefficients for amino acid classes, including neutral, polar, hydrophobic, and charged species, were measured and compared with values for other ionic solutes such as phosphate. The rates of efflux of glycine, lysine, phenylalanine, serine and tryptophan were determined after they were passively entrapped in large unilamellar vesicles (LUVs) composed of egg phosphatidylcholine (EPC) or dimyristoylphosphatidylcholine (DMPC). The following permeability coefficients were obtained for: glycine, 5.7 x 10(-12) cm s-1 (EPC), 2.0 x 10(-11) cm s-1 (DMPC); serine, 5.5 x 10(-12) cm s-1 (EPC), 1.6 x 10(-11) cm s-1 (DMPC); lysine, 5.1 x 10(-12) cm s-1 (EPC), 1.9 x 10(-11) cm s-1 (DMPC); tryptophan, 4.1 x 10(-10) cm s-1 (EPC); and phenylalanine, 2.5 x 10(-10) cm s-1 (EPC). Decreasing lipid chain length increased permeability slightly, while variations in pH had only minor effects on the permeability coefficients of the amino acids tested. Phosphate permeability was in the range of 10(-12)-10(-13) cm s-1 depending on the pH of the medium. The values for the polar and charged amino acids were surprisingly similar to those previously measured for monovalent cations such as sodium and potassium, which are in the range of 10(-12)-10(-13) cm s-1, depending on conditions and the lipid species used. This observation suggests that the permeation rates for the neutral, polar and charged amino acids are controlled by bilayer fluctuations and transient defects, rather than partition coefficients and Born energy barriers. The results are relevant to the permeation of certain peptides into lipid bilayers during protein translocation and membrane biogenesis.

  16. Studies on preparation of medium fat liquid dairy whitener from buffalo milk employing ultrafiltration process.

    PubMed

    Khatkar, Sunil Kumar; Gupta, Vijay Kumar; Khatkar, Anju Boora

    2014-09-01

    A study was conducted to develop good quality medium fat liquid dairy whitener from buffalo milk employing ultrafiltration (UF) process. The buffalo skim milk was UF concentrated to 4.05 to 4.18 (23.63 ± 0.30 % TS) fold and standardized to 10 % fat (on Dry Matter Basis) (i.e. formulation) and homogenized at 175.76 kg/cm(2). The addition of 0.4 % mixture of monosodium and disodium phosphate (2:1 w/w) improved the heat stability of homogenized formulation to an optimum of 66 min. The bland flavour of homogenized formulation with added 0.4 % mixture of monosodium phosphate and disodium phosphate (2:1 w/w) and 18 % sugar (on DMB) (i.e. medium fat liquid dairy whitener) was improved significantly (P < 0.01) with the addition of 0.2 % potassium chloride, but heat stability of medium fat liquid dairy whitener got reduced substantially (i.e. 19 min). With subsequent heat treatment to 85 °C for 5 min, heat stability of medium fat liquid dairy whitener got improved to reasonable level of 27 min. Whitening ability in terms of L* value of medium fat liquid dairy whitener in both tea and coffee was significantly (P < 0.01) better when homogenized at 175.76 kg/cm(2) vis-à-vis 140.61 kg/cm(2). Standardized medium fat liquid dairy whitener had significantly (P < 0.01) greater protein content (i.e. approximately 2.43 times) compared to market dairy whitener samples. At 2 % solids level, standardized medium fat liquid dairy whitener in tea/coffee fetched significantly (P < 0.01) better sensory attributes and instrumental whitening ability compared to market sample at 3 % solids level. There could be clear 33 % solids quantity saving in case of developed product compared to market dairy whitener sample.

  17. A new method to effectively and rapidly generate neurons from SH-SY5Y cells.

    PubMed

    Yang, HongNa; Wang, Jing; Sun, JinHua; Liu, XiaoDun; Duan, Wei-Ming; Qu, TingYu

    2016-01-01

    It is well known that neurons differentiated from SH-SY5Y cells can serve as cell models for neuroscience research; i.e., neurotoxicity and tolerance to morphine in vitro. To differentiate SH-SY5Y cells into neurons, RA (retinoic acid) is commonly used to produce the inductive effect. However, the percentage of neuronal cells produced from SH-SY5Y cells is low, either from the use of RA treatment alone or from the combined application of RA and other chemicals. In the current study, we used CM-hNSCs (conditioned medium of human neural stem cells) as the combinational inducer with RA to prompt neuronal differentiation of SH-SY5Y cells. We found that neuronal differentiation was improved and that neurons were greatly increased in the differentiated SH-SY5Y cells using a combined treatment of CM-hNSCs and RA compared to RA treatment alone. The neuronal percentage was higher than 80% (about 88%) on the 3rd day and about 91% on the 7th day examined after a combined treatment with CM-hNSCs and RA. Cell maturation and neurite growth of these neuronal cells were also improved. In addition, the use of CM-hNSCs inhibited the apoptosis of RA-treated SH-SY5Y cells in culture. We are the first to report the use of CM-hNSCs in combination with RA to induce neuronal differentiation of RA-treated SH-SY5Y cells. Our method can rapidly and effectively promote the neuronal production of SH-SY5Y cells in culture conditions. Copyright © 2015 Elsevier Ireland Ltd. All rights reserved.

  18. Paracrine-mediated osteoclastogenesis by the osteosarcoma MG63 cell line: is RANKL/RANK signalling really important?

    PubMed

    Costa-Rodrigues, J; Teixeira, C A; Fernandes, M H

    2011-08-01

    Although in the past little attention has been paid to the influence of osteosarcoma cells in osteoclast function, recent studies suggest a close relationship between osteosarcoma aggressiveness and osteoclastic activity. The present study addresses the paracrine effects of MG63 cells, a human osteosarcoma-derived cell line, on the differentiation of peripheral blood osteoclast precursor cells (PBMC). PBMC were cultured for 21 days in the presence of conditioned media from MG63 cell cultures (CM) collected at 48 h (CM_MG1), 7 days (CM_MG2) and 14 days (CM_MG3). MG63 cell cultures displayed the expression of ALP and BMP-2 and, also, the osteoclastogenic genes M-CSF and RANKL, although with a low expression of RANKL. PBMC cultures supplemented with CM presented an evident osteoclastogenic behavior, which was dependent on the culture period of the MG63 cells. The inductive effect appeared to be more relevant for the differentiation and activation genes, c-myc and c-src, and lower for genes associated with osteoclast function. In addition, PBMC cultures displayed increased functional parameters, including calcium phosphate resorbing activity. Assessment of the PBMC cultures in the presence of U0126, PDTC, and indomethacin suggested that in addition to MEK and NFkB pathways, other signaling mechanisms, probably not involving RANKL/RANK interaction, might be activated in the presence of conditioned medium from MG63. In conclusion, MG63 cell line appears to induce a significant paracrine-mediated osteoclastogenic response. Understanding the mechanisms underlying the interaction of osteosarcoma cells and osteoclasts may contribute to the development of new potential approaches in the treatment of such bone metabolic diseases.

  19. Follow-up treatment effects of contingency management and motivational interviewing on substance use: A meta-analysis.

    PubMed

    Sayegh, Caitlin S; Huey, Stanley J; Zara, Erica J; Jhaveri, Kinnari

    2017-06-01

    Motivation is an integral factor in substance use treatment and long-term recovery. However, it is unclear what role intrinsic and extrinsic motivation play across different treatment modalities. A meta-analysis (N = 84) was performed to estimate the pooled effect size of Motivational Interviewing (MI; primarily targeting intrinsic motivation) and contingency management (CM; primarily targeting extrinsic motivation) at different follow-up periods. Collapsed across all substance types, CM had a significant effect at 3-month follow-up, only. In contrast, MI had a significant effect at 6-month follow-up, only. CM had small and medium effects on multiple substances at 3-month follow-up (i.e., tobacco, marijuana, stimulants, polysubstances), but not at 6-month follow-up. MI had 1 significant medium effect at 3-month follow-up (i.e., marijuana), but several significant small effects at 6-month follow-up (i.e., alcohol, tobacco, polysubstances). This meta-analysis suggests that both CM and MI promote reductions in a range of substances, even several months after the intervention concludes. Further, these results provide some evidence that extrinsically focused CM may produce medium follow-up effects in the short run, but intrinsically focused MI may produce small but durable follow-up effects. However, this interpretation is complicated by the differences between the MI and CM studies that preclude statistical tests comparing effect sizes, and few studies assessed motivation itself. Future researchers should investigate how motivational dynamics impact lasting outcomes in substance use treatment. (PsycINFO Database Record (c) 2017 APA, all rights reserved).

  20. Specific yield - laboratory experiments showing the effect of time on column drainage

    USGS Publications Warehouse

    Prill, Robert C.; Johnson, A.I.; Morris, Donald Arthur

    1965-01-01

    The increasing use of ground water from many major aquifers in the United States has required a more thorough understanding of gravity drainage, or specific yield. This report describes one phase of specific yield research by the U.S. Geological Survey's Hydrologic Laboratory in cooperation with the California Department of Water Resources. An earlier phase of the research concentrated on the final distribution of moisture retained after drainage of saturated columns of porous media. This report presents the phase that concentrated on the distribution of moisture retained in similar columns after drainage for various periods of time. Five columns, about 4 cm in diameter by 170 cm long, were packed with homogenous sand of very fine, medium, and coarse sizes, and one column was packed with alternating layers of coarse and medium sand. The very fine materials were more uniform in size range than were the medium materials. As the saturated columns drained, tensiometers installed throughout the length recorded changes in moisture tension. The relation of tension to moisture content, determined for each of the materials, was then used to convert the tension readings to moisture content. Data were then available on the distribution of retained moisture for different periods of drainage from 1 to 148 hours. Data also are presented on the final distribution of moisture content by weight and volume and on the degree of saturation. The final zone of capillary saturation was approximately 12 cm for coarse sand, 13 cm for medium sand, and 52 cm for very fine sand. The data showed these zones were 92 to 100 percent saturated. Most of the outflow from the columns occurred in the earlier hours of drainage--90 percent in 1 hour for the coarse materials, 50 percent for the medium, and 60 percent for the very fine. Although the largest percentage of the specific yield was reached during the early hours of .drainage, this study amply demonstrates that a very long time would be required to reach drainage equilibrium. In the layered columns the middle (medium sand) layer functioned as a hanging water column accelerating the drainage of the overlying coarse-sand layer. After the middle layer started to drain, the moisture distribution as retained in all three layers showed trends similar to that obtained when the same materials were tested in homogenous columns.

  1. 75 FR 51182 - Atlantic Highly Migratory Species; Atlantic Bluefin Tuna Fisheries

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-08-19

    ... one to three large medium or giant BFT for the September, October- November, and December time periods... the daily retention limit of large medium and giant BFT over a range of zero to a maximum of three per... limit of one large medium or giant BFT (measuring 73 inches (185 cm) curved fork length (CFL) or greater...

  2. Amplification and gas-dynamic parameters of the active oxygen-iodine medium produced by an ejector nozzle unit

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

    Zagidullin, M V; Nikolaev, V D; Svistun, M I

    2001-08-31

    The gain, the temperature, and the absolute velocity of the supersonic active oxygen-iodine medium produced by an ejector nozzle unit were determined by the technique of high-resolution diode laser spectroscopy. The gain in the active medium is formed at less than 44 mm from the nozzle unit for an absolute flow velocity {nu} {approx} 600 m s{sup -1}. Upon dilution of oxygen by primary nitrogen in the ratio of 1 : 6.9, the gain of the active medium amounts to 7x10{sup -3} cm{sup -1}, the temperature of the active medium to 200 K, the absolute flow velocity to 580 mmore » s{sup -1}, and the pressure to 58 Torr. As the dilution is increased to 1 : 13.5, the gain reduces to 4.5x10{sup -3} cm{sup -1}, the temperature lowers to 180 K, the velocity of the active medium increases to 615 m s{sup -1}, and the pressure increases to 88 Torr. The increase in the initial content of water vapour in the oxygen flow results in an increase in the temperature and a decrease in the gain of the active medium. (active media)« less

  3. Grey water treatment in urban slums by a filtration system: optimisation of the filtration medium.

    PubMed

    Katukiza, A Y; Ronteltap, M; Niwagaba, C B; Kansiime, F; Lens, P N L

    2014-12-15

    Two uPVC columns (outer diameter 160 cm, internal diameter 14.6 cm and length 100 cm) were operated in parallel and in series to simulate grey water treatment by media based filtration at unsaturated conditions and constant hydraulic loading rates (HLR). Grey water from bathroom, laundry and kitchen activities was collected from 10 households in the Bwaise III slum in Kampala (Uganda) in separate containers, mixed in equal proportions followed by settling, prior to transferring the influent to the tanks. Column 1 was packed with lava rock to a depth of 60 cm, while column 2 was packed with lava rock (bottom 30 cm) and silica sand, which was later replaced by granular activated carbon (top 30 cm) to further investigate nutrient removal from grey water. Operating the two filter columns in series at a HLR of 20 cm/day resulted in a better effluent quality than at a higher (40 cm/day) HLR. The COD removal efficiencies by filter columns 1 and 2 in series amounted to 90% and 84% at HLR of 20 cm/day and 40 cm/day, respectively. TOC and DOC removal efficiency amounted to 77% and 71% at a HLR of 20 cm/day, but decreased to 72% and 67% at a HLR of 40 cm/day, respectively. The highest log removal of Escherichia coli, Salmonella sp. and total coliforms amounted to 3.68, 3.50 and 3.95 at a HLR of 20 cm/day respectively. The overall removal of pollutants increased with infiltration depth, with the highest pollutant removal efficiency occurring in the top 15 cm layer. Grey water pre-treatment followed by double filtration using coarse and fine media has the potential to reduce the grey water pollution load in slum areas by more than 60%. Copyright © 2014 Elsevier Ltd. All rights reserved.

  4. Micropropagation of Ajuga bracteosa, a medicinal herb.

    PubMed

    Kaul, Shivanee; Das, Sandip; Srivastava, P S

    2013-04-01

    For conservation and genetic transformation, a successful in vitro micropropagation protocol for Ajuga bracteosa, a medicinal herb has been established for the first time. MS medium supplemented with IAA (2 mg/L) and BA (5 mg/L) induced 100 % shoot regeneration with an average of 41.4 shoots of 8.4 cm per culture. Excised in vitro shoots when transferred to MS + IBA (0.5 mg/L) produced 20 roots/shoot of 20.2 cm average length in 100 % cultures. Of the three explants, leaf, petiole and root, leaf displayed quickest response followed by petiole while root was the slowest. Hardening of plantlets was achieved with 82 % survival. The hardened plants were maintained in pots with garden soil under controlled (Temp. 25 ± 2 °C) conditions. RAPD exhibited genetic fidelity with 100 % monomorphism in regenerants.

  5. Chemistry of nitrile anions in the interstellar medium

    NASA Astrophysics Data System (ADS)

    Carles, S.; Le Garrec, J.-L.; Guillemin, J.-C.; Biennier, L.

    2015-12-01

    Despite the extreme conditions of temperature (down to 10K) and density (down to 100 molecules/cm3), the giant molecular clouds and the circumstellar envelopes present a rich and complex chemistry. To date, more than 180 molecules have been detected in the InterStellar Medium (ISM) with a large abundance of nitriles (RC≡N). In addition, several anions have been recently observed in this medium: C4H¯, C6H¯, C8H¯, CN¯, C3N¯ and C5N¯. These last species should play a key role in the molecular growth towards complexity. To explore this hypothesis, their reactivity must be studied in the laboratory. The FALP-MS and the CRESU experimental apparatuses of the Rennes University are able to measure absolute rate coefficient of various chemical reactions, including the ion - molecule reactions, in gas phase at low temperature (from 300K for the FALP-MS down to 15K for the CRESU). Therefore, these experimental tools are particularly adapted to the kinetic studies of reactions potentially involved in the Interstellar Medium. One of the difficulties encountered in experiments with anions is their generation. We describe here the formation of the CN¯ and C3N¯ anions by dissociative electron attachment on the molecular precursors BrCN and BrC3N.

  6. DICKE’S SUPERRADIANCE IN ASTROPHYSICS. I. THE 21 cm LINE

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

    Rajabi, Fereshteh; Houde, Martin

    We have applied the concept of superradiance introduced by Dicke in 1954 to astrophysics by extending the corresponding analysis to the magnetic dipole interaction characterizing the atomic hydrogen 21 cm line. Although it is unlikely that superradiance could take place in thermally relaxed regions and that the lack of observational evidence of masers for this transition reduces the probability of detecting superradiance, in situations where the conditions necessary for superradiance are met (close atomic spacing, high velocity coherence, population inversion, and long dephasing timescales compared to those related to coherent behavior), our results suggest that relatively low levels of populationmore » inversion over short astronomical length-scales (e.g., as compared to those required for maser amplification) can lead to the cooperative behavior required for superradiance in the interstellar medium. Given the results of our analysis, we expect the observational properties of 21 cm superradiance to be characterized by the emission of high-intensity, spatially compact, burst-like features potentially taking place over short periods ranging from minutes to days.« less

  7. EMMPRIN Is Secreted by Human Uterine Epithelial Cells in Microvesicles and Stimulates Metalloproteinase Production by Human Uterine Fibroblast Cells

    PubMed Central

    Dayger, C. A.; Mehrotra, P.; Belton, R. J.; Nowak, R. A.

    2012-01-01

    Endometrial remodeling is a physiological process involved in the gynecological disease, endometriosis. Tissue remodeling is directed by uterine fibroblast production of matrix metalloproteinases (MMPs). Several MMPs are regulated directly by the protein extracellular matrix metalloproteinase inducer (EMMPRIN) and also by proinflammatory cytokines such as interleukin (IL)1-α/β. We hypothesized that human uterine epithelial cells (HESs) secrete intact EMMPRIN to stimulate MMPs. Microvesicles from HES cell-conditioned medium (CM) expressed intact EMMPRIN protein. Treatment of HES cells with estradiol or phorbyl 12-myristate-13-acetate increased the release of EMMPRIN-containing microvesicles. The HES CM stimulated MMP-1, -2, and -3 messenger RNA levels in human uterine fibroblasts (HUFs) and EMMPRIN immunodepletion from HES-cell concentrated CM reduced MMP stimulation (P < .05). Treatment of HUF cells with low concentrations of IL-1β/α stimulated MMP production (P < .05). These results indicate that HES cells regulate MMP production by HUF cells by secretion of EMMPRIN, in response to ovarian hormones, proinflammatory cytokines as well as activation of protein kinase C. PMID:22729071

  8. EMMPRIN is secreted by human uterine epithelial cells in microvesicles and stimulates metalloproteinase production by human uterine fibroblast cells.

    PubMed

    Braundmeier, A G; Dayger, C A; Mehrotra, P; Belton, R J; Nowak, R A

    2012-12-01

    Endometrial remodeling is a physiological process involved in the gynecological disease, endometriosis. Tissue remodeling is directed by uterine fibroblast production of matrix metalloproteinases (MMPs). Several MMPs are regulated directly by the protein extracellular matrix metalloproteinase inducer (EMMPRIN) and also by proinflammatory cytokines such as interleukin (IL)1-α/β. We hypothesized that human uterine epithelial cells (HESs) secrete intact EMMPRIN to stimulate MMPs. Microvesicles from HES cell-conditioned medium (CM) expressed intact EMMPRIN protein. Treatment of HES cells with estradiol or phorbyl 12-myristate-13-acetate increased the release of EMMPRIN-containing microvesicles. The HES CM stimulated MMP-1, -2, and -3 messenger RNA levels in human uterine fibroblasts (HUFs) and EMMPRIN immunodepletion from HES-cell concentrated CM reduced MMP stimulation (P < .05). Treatment of HUF cells with low concentrations of IL-1β/α stimulated MMP production (P < .05). These results indicate that HES cells regulate MMP production by HUF cells by secretion of EMMPRIN, in response to ovarian hormones, proinflammatory cytokines as well as activation of protein kinase C.

  9. The secretome of periodontal ligament stem cells from MS patients protects against EAE

    PubMed Central

    Rajan, Thangavelu Soundara; Giacoppo, Sabrina; Diomede, Francesca; Ballerini, Patrizia; Paolantonio, Michele; Marchisio, Marco; Piattelli, Adriano; Bramanti, Placido; Mazzon, Emanuela; Trubiani, Oriana

    2016-01-01

    Manipulation of stem cells or stem cells-derived secretome has emerged as a novel alternative therapeutic option for multiple sclerosis (MS). Here we show that human periodontal ligament stem cells (hPDLSCs)-derived conditioned medium (hPDLSCs-CM) and purified exosomes/microvesicles (hPDLSCs-EMVs) obtained from Relapsing Remitting (RR)-MS patients and healthy donors block experimental autoimmune encephalomyelitis (EAE), a mouse model of MS, by inducing anti-inflammatory and immunosuppressive effects in spinal cord and spleen, and reverse disease progression by restoring tissue integrity via remyelination in the spinal cord. We show that hPDLSCs-CM and hPDLSCs-EMVs reduce pro-inflammatory cytokines IL-17, IFN-γ, IL-1β, IL-6, TNF-α, and induce anti-inflammatory IL-10. In addition, apoptosis related STAT1, p53, Caspase 3, and Bax expressions were attenuated. Our findings unravel the immunosuppressive effects of hPDLSCs-CM and hPDLSCs-EMVs in EAE mice, and suggest simple alternative autologous source for patient-customized cell-free targeting treatment in MS patients. PMID:27924938

  10. A large bubble around the Crab Nebula

    NASA Technical Reports Server (NTRS)

    Romani, Roger W.; Reach, William T.; Koo, Bon Chul; Heiles, Carl

    1990-01-01

    IRAS and 21 cm observations of the interstellar medium around the Crab nebula show evidence of a large bubble surrounded by a partial shell. If located at the canonical 2 kpc distance of the Crab pulsar, the shell is estimated to have a radius of about 90 pc and to contain about 50,000 solar masses of swept-up gas. The way in which interior conditions of this bubble can have important implications for observations of the Crab are described, and the fashion in which presupernova evolution of the pulsar progenitor has affected its local environment is described.

  11. The Differentiation of Human Adipose-Derived Stem Cells towards a Urothelium-Like Phenotype In Vitro and the Dynamic Temporal Changes of Related Cytokines by Both Paracrine and Autocrine Signal Regulation

    PubMed Central

    Zhou, Zhe; Zhang, Ke; Zhou, Juan; Zhao, Yang; Wang, Zhong; Lu, Mu-jun

    2014-01-01

    Purpose To investigate the differentiation ability of human adipose-derived stem cells (ASCs) towards urothelium-like cells in vitro and the dynamic changes of related cytokines and cytokine receptors in the culture medium. Materials and Methods The ASCs were induced using both conditioned media (CM) and the transwell co-culture system with an immortalized urothelium cell line (SV-HUC-1,HUC) for 21 days. Protein and mRNA expression of the mature urothelium specific markers uroplakin-IA (UP-1A) and uroplakin-II (UP-II) were detected by immunofluorescence and quantitative real-time PCR, respectively. Array detection was used to screen 41 cytokines and receptors in the upper medium of urothelium, non-induced ASCs and urothelium-induced ASCs at three time points, early (12 hours), intermediate (7 days) and late (21 days). Results After induction for 7 days, the ASCs grown in both CM and transwell co-culture system expressed uroplakin-IA (13.54±2.00%; 17.28±1.84%) and uroplakin-II (19.49±1.73%; 13.98±1.47%). After induction for 21 days, ASCs grown in co-culture had significantly increased expression of uroplakin-IA (48.03±1.25%; 49.57±2.85%) and uroplakin-II (45.38±2.50%; 46.58±1.95%). In the upper medium of urothelium, 28 cytokines and 8 cytokine receptors had significantly higher expression than the counterpart of non-induced ASCs. After 7 days induction, the expression of 22 cytokines and 8 cytokine receptors was significantly elevated in the upper medium of induced ASCs compared to non-induced ASCs. At the early and intermediate time points, ASCs secreted high levels of relative cytokines and soluble receptors, but their expressions decreased significantly at the late time point. Conclusion The adipose-derived stem cells have the potential to be differentiated into urothelium-like cells in vitro by both CM and transwell co-culture system with mature urothelium. Numerous cytokines and receptors were involved in the differentiation process with dynamic temporal changes by both paracrine and autocrine signal regulation. Further studies should be carried out to determine the detailed mechanism of cytokines and receptors and to enhance the urothelium differentiation efficiency of ASCs. PMID:24752317

  12. The differentiation of human adipose-derived stem cells towards a urothelium-like phenotype in vitro and the dynamic temporal changes of related cytokines by both paracrine and autocrine signal regulation.

    PubMed

    Zhang, Ming; Xu, Ming-Xi; Zhou, Zhe; Zhang, Ke; Zhou, Juan; Zhao, Yang; Wang, Zhong; Lu, Mu-Jun

    2014-01-01

    To investigate the differentiation ability of human adipose-derived stem cells (ASCs) towards urothelium-like cells in vitro and the dynamic changes of related cytokines and cytokine receptors in the culture medium. The ASCs were induced using both conditioned media (CM) and the transwell co-culture system with an immortalized urothelium cell line (SV-HUC-1,HUC) for 21 days. Protein and mRNA expression of the mature urothelium specific markers uroplakin-IA (UP-1A) and uroplakin-II (UP-II) were detected by immunofluorescence and quantitative real-time PCR, respectively. Array detection was used to screen 41 cytokines and receptors in the upper medium of urothelium, non-induced ASCs and urothelium-induced ASCs at three time points, early (12 hours), intermediate (7 days) and late (21 days). After induction for 7 days, the ASCs grown in both CM and transwell co-culture system expressed uroplakin-IA (13.54±2.00%; 17.28±1.84%) and uroplakin-II (19.49±1.73%; 13.98±1.47%). After induction for 21 days, ASCs grown in co-culture had significantly increased expression of uroplakin-IA (48.03±1.25%; 49.57±2.85%) and uroplakin-II (45.38±2.50%; 46.58±1.95%). In the upper medium of urothelium, 28 cytokines and 8 cytokine receptors had significantly higher expression than the counterpart of non-induced ASCs. After 7 days induction, the expression of 22 cytokines and 8 cytokine receptors was significantly elevated in the upper medium of induced ASCs compared to non-induced ASCs. At the early and intermediate time points, ASCs secreted high levels of relative cytokines and soluble receptors, but their expressions decreased significantly at the late time point. The adipose-derived stem cells have the potential to be differentiated into urothelium-like cells in vitro by both CM and transwell co-culture system with mature urothelium. Numerous cytokines and receptors were involved in the differentiation process with dynamic temporal changes by both paracrine and autocrine signal regulation. Further studies should be carried out to determine the detailed mechanism of cytokines and receptors and to enhance the urothelium differentiation efficiency of ASCs.

  13. The structure of the Cepheus E protostellar outflow: The jet, the bowshock, and the cavity

    NASA Astrophysics Data System (ADS)

    Lefloch, B.; Gusdorf, A.; Codella, C.; Eislöffel, J.; Neri, R.; Gómez-Ruiz, A. I.; Güsten, R.; Leurini, S.; Risacher, C.; Benedettini, M.

    2015-09-01

    Context. Protostellar outflows are a crucial ingredient of the star-formation process. However, the physical conditions in the warm outflowing gas are still poorly known. Aims: We present a multi-transition, high spectral resolution CO study of the outflow of the intermediate-mass Class 0 protostar Cep E-mm. The goal is to determine the structure of the outflow and to constrain the physical conditions of the various components in order to understand the origin of the mass-loss phenomenon. Methods: We have observed the J = 12-11, J = 13-12, and J = 16-15 CO lines at high spectral resolution with SOFIA/GREAT and the J = 5-4, J = 9-8, and J = 14-13 CO lines with HIFI/Herschel towards the position of the terminal bowshock HH377 in the southern outflow lobe. These observations were complemented with maps of CO transitions obtained with the IRAM 30 m telescope (J = 1-0, 2-1), the Plateau de Bure interferometer (J = 2-1), and the James Clerk Maxwell Telescope (J = 3-2, 4-3). Results: We identify three main components in the protostellar outflow: the jet, the cavity, and the bowshock, with a typical size of 1.7″ × 21″, 4.5″, and 22″ × 10″, respectively. In the jet, the emission from the low-J CO lines is dominated by a gas layer at Tkin = 80-100 K, column density N(CO) = 9 × 1016 cm-2, and density n(H2) = (0.5-1) × 105 cm-3; the emission of the high-J CO lines arises from a warmer (Tkin = 400-750 K), denser (n(H2) = (0.5-1) × 106 cm-3), lower column density (N(CO) = 1.5 × 1016 cm-2) gas component. Similarly, in the outflow cavity, two components are detected: the emission of the low-J lines is dominated by a gas layer of column density N(CO) = 7 × 1017 cm-2 at Tkin = 55-85 K and density in the range (1-8) × 105 cm-3; the emission of the high-J lines is dominated by a hot, denser gas layer with Tkin = 500-1500K, n(H2) = (1-5) × 106 cm-3, and N(CO) = 6 × 1016 cm-2. A temperature gradient as a function of the velocity is found in the high-excitation gas component. In the terminal bowshock HH377, we detect gas of moderate excitation, with a temperature in the range Tkin ≈ 400-500 K, density n(H2) ≃ (1 -2) × 106 cm-3 and column density N(CO) = 1017 cm-2. The amounts of momentum carried away in the jet and in the entrained ambient medium are similar. Comparison with time-dependent shock models shows that the hot gas emission in the jet is well accounted for by a magnetized shock with an age of 220-740 yr propagating at 20-30 km s-1 in a medium of density n(H2) = (0.5-1) × 105 cm-3, consistent with that of the bulk material. Conclusions: The Cep E protostellar outflow appears to be a convincing case of jet bowshock driven outflow. Our observations trace the recent impact of the protostellar jet into the ambient cloud, produing a non-stationary magnetized shock, which drives the formation of an outflow cavity. Appendices are available in electronic form at http://www.aanda.org

  14. Dissolved oxygen concentration in the medium during cell culture: Defects and improvements.

    PubMed

    Zhang, Kuan; Zhao, Tong; Huang, Xin; He, Yunlin; Zhou, Yanzhao; Wu, Liying; Wu, Kuiwu; Fan, Ming; Zhu, Lingling

    2016-03-01

    In vitro cell culture has provided a useful model to study the effects of oxygen on cellular behavior. However, it remains unknown whether the in vitro operations themselves affect the medium oxygen levels and the living states of cells. In addition, a prevailing controversy is whether reactive oxygen species (ROS) production is induced by continuous hypoxia or reoxygenation. In this study, we have measured the effects of different types of cell culture containers and the oxygen environment where medium replacement takes place on the actual oxygen tension in the medium. We found that the deviations of oxygen concentrations in the medium are much greater in 25-cm(2) flasks than in 24-well plates and 35-mm dishes. The dissolved oxygen concentrations in the medium were increased after medium replacement in normoxia, but remained unchanged in glove boxes in which the oxygen tension remained at a low level (11.4, 5.7, and 0.5% O2 ). We also found that medium replacement in normoxia increased the number of ROS-positive cells and reduced the cell viability; meanwhile, medium replacement in a glove box did not produce the above effects. Therefore, we conclude that the use of 25-cm(2) flasks should be avoided and demonstrate that continuous hypoxia does not produce ROS, whereas the reoxygenation that occurs during the harvesting of cells leads to ROS and induces cell death. © 2015 International Federation for Cell Biology.

  15. 18 years' experience with high dose rate strontium-90 brachytherapy of small to medium sized posterior uveal melanoma.

    PubMed

    van Ginderdeuren, R; van Limbergen, E; Spileers, W

    2005-10-01

    To analyse local tumour control, radiation related complications, visual acuity, enucleation rate, and survival after brachytherapy of small to medium sized choroidal melanoma (CM) with a high dose rate (HDR) strontium-90 (Sr-90) applicator. From 1983 until 2000, 98 eyes with CM were treated with Sr-90 brachytherapy. The main outcome measures were actuarial rates of the patients' survival, ocular conservation rate, tumour regression, complication rates, and preservation of visual acuity. End point rates were estimated using Kaplan-Meier analysis. The median follow up time was 6.7 years (0.5-18.8 years). Actuarial melanoma free patient survival rate was 85% (SE 4.8%) after 18 years. Actuarial rate of ocular conservation and complete tumour regression was 90% (SE 3.8%) after 15 years. In 93% local tumour control was achieved, 88% showed a stable scar. Recurrence of the tumour on the border caused enucleation of six eyes (7%). In three cases (4%) retinal detachment was the end point. No cases of optic atrophy or of sight impairing retinopathy outside the treated area were found. Actuarial rate of preservation of visual acuity of 1/10 was 65% at 5 years and 45% at 15 years of follow up (SE 5.9% and 8.8%). Sr-90 brachytherapy is as effective as iodine or ruthenium brachytherapy for small to medium sized CM but causes fewer complications. The preservation of vision is better than with all other described radioisotopes. HDR Sr-90 brachytherapy can therefore safely be recommended for small to medium sized CM.

  16. 18 Years’ experience with high dose rate strontium-90 brachytherapy of small to medium sized posterior uveal melanoma

    PubMed Central

    van Ginderdeuren, R; van Limbergen, E; Spileers, W

    2005-01-01

    Aim: To analyse local tumour control, radiation related complications, visual acuity, enucleation rate, and survival after brachytherapy of small to medium sized choroidal melanoma (CM) with a high dose rate (HDR) strontium-90 (Sr-90) applicator. Methods: From 1983 until 2000, 98 eyes with CM were treated with Sr-90 brachytherapy. The main outcome measures were actuarial rates of the patients’ survival, ocular conservation rate, tumour regression, complication rates, and preservation of visual acuity. End point rates were estimated using Kaplan-Meier analysis. Results: The median follow up time was 6.7 years (0.5–18.8 years). Actuarial melanoma free patient survival rate was 85% (SE 4.8%) after 18 years. Actuarial rate of ocular conservation and complete tumour regression was 90% (SE 3.8%) after 15 years. In 93% local tumour control was achieved, 88% showed a stable scar. Recurrence of the tumour on the border caused enucleation of six eyes (7%). In three cases (4%) retinal detachment was the end point. No cases of optic atrophy or of sight impairing retinopathy outside the treated area were found. Actuarial rate of preservation of visual acuity of 1/10 was 65% at 5 years and 45% at 15 years of follow up (SE 5.9% and 8.8%). Conclusions: Sr-90 brachytherapy is as effective as iodine or ruthenium brachytherapy for small to medium sized CM but causes fewer complications. The preservation of vision is better than with all other described radioisotopes. HDR Sr-90 brachytherapy can therefore safely be recommended for small to medium sized CM. PMID:16170122

  17. Early tumor growth in metastatic organs influenced by the microenvironment is an important factor which provides organ specificity of colon cancer metastasis.

    PubMed

    Hara, Y; Ogata, Y; Shirouzu, K

    2000-12-01

    We have previously demonstrated that liver metastases in nude mice and lung metastases in nude rats occurred specifically, when KM12SM human colon carcinoma cells were inoculated orthotopically into the cecal wall of nude mice and rats. To clarify the relationship between the tumor growth potential in the metastatic organs and the metastatic organ preference in these two metastatic models, we have evaluated the in vitro cell growth activities affected by the organ conditioned medium (CM) from the liver and lung, and the in vivo growth activities of the ectopic implanted tumors in the liver and lung. The tumorigenicity of the ectopic implanted tumors was 100% in mouse liver, 33% in rat liver, 50% in mouse lung, and 75% in rat lung. The crude liver CM of the animals showed inhibitory activities for KM12SM cell growth in a dosage-dependent manner, and the crude lung CM stimulated KM12SM cell growth. The liver CM of nude mice inhibited the KM12SM cell growth more strongly compared with the CM of nude rats, and the lung CM of nude rats was more strongly stimulated compared with the CM of nude mice. The liver CM of nude mice had non-heparin binding factors, which stimulated or inhibited KM12SM cell growth, in a molecular weight range of 50 to 100 kDa. By contrast, the liver CM of nude rats showed no growth stimulating activity for KM12SM cells. These results suggest that the metastatic organ specificity of KM12SM cells may depend on the early tumor growth influenced by the microenvironment in metastatic organs.

  18. Non-cladding optical fiber is available for detecting blood or liquids.

    PubMed

    Takeuchi, Akihiro; Miwa, Tomohiro; Shirataka, Masuo; Sawada, Minoru; Imaizumi, Haruo; Sugibuchi, Hiroyuki; Ikeda, Noriaki

    2010-10-01

    Serious accidents during hemodialysis such as an undetected large amount of blood loss are often caused by venous needle dislodgement. A special plastic optical fiber with a low refractive index was developed for monitoring leakage in oil pipelines and in other industrial fields. To apply optical fiber as a bleeding sensor, we studied optical effects of soaking the fiber with liquids and blood in light-loss experimental settings. The non-cladding optical fiber that was used was the fluoropolymer, PFA fiber, JUNFLON™, 1 mm in diameter and 2 m in length. Light intensity was studied with an ordinary basic circuit with a light emitting source (880 nm) and photodiode set at both terminals of the fiber under certain conditions: bending the fiber, soaking with various mediums, or fixing the fiber with surgical tape. The soaking mediums were reverse osmosis (RO) water, physiological saline, glucose, porcine plasma, and porcine blood. The light intensities regressed to a decaying exponential function with the soaked length. The light intensity was not decreased at bending from 20 to 1 cm in diameter. The more the soaked length increased in all mediums, the more the light intensity decreased exponentially. The means of five estimated exponential decay constants were 0.050±0.006 standard deviation in RO water, 0.485±0.016 in physiological saline, 0.404±0.022 in 5% glucose, 0.503±0.038 in blood (Hct 40%), and 0.573±0.067 in plasma. The light intensity decreased from 5 V to about 1.5 V above 5 cm in the soaked length in mediums except for RO water and fixing with surgical tape. We confirmed that light intensity significantly and exponentially decreased with the increased length of the soaked fiber. This phenomena could ideally, clinically be applied to a bleed sensor.

  19. The influence of oxygen and hydrostatic pressure on articular chondrocytes and adherent bone marrow cells in vitro.

    PubMed

    Scherer, Katrin; Schünke, Michael; Sellckau, Roland; Hassenpflug, Joachim; Kurz, Bodo

    2004-01-01

    Tissue engineering of articular cartilage from chondrocytes or stem cells is considered to be a potential aspect in the treatment of cartilage defects. In order to optimize culture conditions the influence of low oxygen tension (5%) - single or in combination with intermittent hydrostatic pressure (HP: 30/2 min on/off loading; 0.2 MPa) - on the biosynthetic activity (sulfate and proline incorporation) of human osteoarthritic chondrocytes cultured on collagen I/III membranes was investigated. Additionally, chondrogenesis from high density or monolayer cultures of bovine adherent bone marrow cells (aBMC) with and without chondrogenic medium supplements (CM) was analyzed by RT-PCR (mRNA expression of aggrecan and collagen type II). We could show that low oxygen tension increases significantly the biosynthesis of collagen I/III membrane-associated chondrocytes and even higher under co-stimulation with HP. While there is no chondrogenesis in monolayer cultures, CM induces expression of cartilage matrix molecules in high density cultures of aBMC which is even increased under the influence of low oxygen tension. Both, low oxygen tension and HP without CM are alone not sufficient stimuli for chondrogenesis. It can be concluded that low oxygen tension and HP might be useful tools in cartilage tissue engineering and that these physico-chemical factors promote but do not induce chondrogenesis under the given conditions.

  20. Mesenchymal stem cells inhibit RANK-RANKL interactions between osteoclasts and Th17 cells via osteoprotegerin activity

    PubMed Central

    Cho, Kyung-Ah; Park, Minhwa; Kim, Yu-Hee; Ryu, Kyung-Ha; Woo, So-Youn

    2017-01-01

    Th17 cells play a critical role in several autoimmune diseases, including psoriasis and psoriatic arthritis (PsA). Psoriasis is a chronic inflammatory skin disease associated with systemic inflammation and comorbidities, such as PsA. PsA develops in nearly 70% of patients with psoriasis, and osteoclasts associated bone erosion is a hallmark of the disease. Thus far, the effect of Th17 cells on osteoclastogenesis via direct cell-to-cell interactions is less understood. In this study, we observed that Th17 cells directly promote osteoclast differentiation and maturation via expression of receptor activator of nuclear factor-κ β ligand (RANKL) in vitro. We investigated the impact of conditioned medium obtained from human palatine tonsil-derived mesenchymal stem cells (T-CM) on the interactions between osteoclasts and Th17 cells. T-CM effectively blunted the RANK-RANKL interaction between the osteoclast precursor cell line RAW 264.7 and Th17 cells via osteoprotegerin (OPG) activity. The frequency of tartrate-resistant acid phosphatase (TRAP)-positive cells in the bone marrow of an imiquimod (IMQ)-induced psoriasis mouse model was decreased following T-CM injection. Therefore, our data provide novel insight into the therapeutic potential of tonsil-derived mesenchymal stem cell-mediated therapy (via OPG production) for the treatment of pathophysiologic processes induced by osteoclasts under chronic inflammatory conditions such as psoriasis. PMID:29137353

  1. The Impact of Biochar on Bioretention Nitrogen Removal and Hydrologic Performance

    NASA Astrophysics Data System (ADS)

    Tian, J.; Jin, J.; Chiu, P.; Guo, M.; Imhoff, P. T.

    2016-12-01

    Poor nitrate removal and substantial land occupation are two factors that limit the application of bioretention facilities. Biochar was evaluated in this study as an amendment of bioretention media to enhance nitrogen removal from runoff as well as improve hydrologic performance. Two pilot-scale bioretention cells (91 cm dia., 1.2 m deep) were constructed in parallel, and both contained 20 cm saturation zone with coarse sand, 76 cm vadose zone with treatment medium, and 5 cm of triple-shredded wood mulch from bottom to top. Treatment medium in the control cell was a mixture of 88% sand, 8% clay, and 4% sawdust by mass, while the biochar cell amended 4% commercial biochar pyrolyzed from Southern Yellow Pine at 550°. Both cells were instrumented with soil moisture sensors, soil potential sensors and temperature sensors. A field infiltration test was conducted in each cell using a tension disc infiltrometer directly on the treatment media to obtain soil hydraulic parameters, then three 24-36-hour tracer tests containing bromide and nitrate pollutant were conducted over a five-month period. Influent, effluent and pore water were continuously sampled for bromide and nitrogen analysis during these tests. In addition, hydrologic performance of the two cells under various conditions of rainfall recurrence interval and duration were simulated using HYDRUS-1D after verification with tracer test data. Results showed that the biochar cell reduced NO3-N concentrations by 30.6-84.7%, while the control cell only reduced NO3-N by -6-43.5%, depending on the storm. Biochar amendment slightly increased the average pH of the vadose zone from 6.3 to 7.3, decreased the average dissolve oxygen content by 43%, and decreased the average oxidation-reduction potential from 22 mV to -115 mV, which contributed to the enhanced nitrate removal. Biochar-amended medium increased saturated hydraulic conductivity by 1.5 times, and increased cell residence time and water retention by 12.6% and 15% respectively during the tracer tests. For a 1-yr interval and 24-hr duration storm simulation, the biochar-amended cell could reduce overflow by 78%, extend the delay of the peak flow by two times and decrease the peak outflow rate by 35%, compared to the control cell.

  2. In vitro cultivation of Hysterothylacium aduncum (Nematoda: Anisakidae) from 3rd-stage larvae to egg-laying adults.

    PubMed

    Iglesias, L; Valero, A; Gálvez, L; Benítez, R; Adroher, F J

    2002-11-01

    This is the first demonstration of the in vitro development of the 3rd-stage larvae (L3) of Hysterothylacium aduncum to the adult. This was achieved in a semi-defined medium that is easy to prepare and to reproduce. The L3, collected from the peritoneal cavity of horse mackerel (Trachurus trachurus), were individually inoculated into RPMI-1640 medium +20% heat-inactivated fetal bovine serum (IFBS). It has been demonstrated that the optimum temperature for development is around 13 degrees C and is stimulated by the presence of 5% CO2 in the growth atmosphere, increasing the percentage moulting to the 4th larval stage (L4) by 1.9-fold (from 44 to 82%) and the average survival of the nematodes by 1.6 times (from 60 to 96 days). When the larvae were grown at different pHs, optimum development occurred at pH 4.0. Under these conditions, all the larvae moulted to the L4 and more than two-thirds transformed to the adult stage--in which 25-30% of the females laid eggs--and reached an average survival of over 4 months. When this medium was supplemented with 1% (w/v) of commercial pepsin, all the larvae reached the adult stage, at least 45% of the females oviposited, laying around 12-fold more eggs per female than in the medium without pepsin. The mean size of the eggs (non-fertilized) obtained was 56.8 x 47.6 microm. The mean length of the adult males obtained was between 3.2 and 5.2 cm and the females were between 3.0 and 6.5 cm. The adult specimens were morphologically identified as Hysterothylacium aducum aduncum. This culture medium (RPMI-1640+20% (v/v) IFBS+1 commercial pepsin, at pH 4.0, 13 degrees C and 5% CO2 in air) could facilitate the identification of at least some of the larvae of the genus Hysterothylacium--and perhaps other anisakids--for which the specific identification and the biological study of these parasites is often difficult.

  3. Cornus sericea

    Treesearch

    Paula M. Pijut

    2004-01-01

    Redosier dogwood is a multistemmed, deciduous, stoloniferous shrub that grows 1 to 3 m tall and 3 m wide or more, often forming dense thickets. Leaves are opposite, simple, ovate to oblonglanceolate, acuminate, rounded at the base, 5 to 12 cm long, 2.5 to 6.4 cm wide, with petioles 1.3 to 2.5 cm long (Dirr 1998, Gleason and Cronquist 1991). Leaves are medium to dark...

  4. Exendin-4 enhances the migration of adipose-derived stem cells to neonatal rat ventricular cardiomyocyte-derived conditioned medium via the phosphoinositide 3-kinase/Akt-stromal cell-derived factor-1α/CXC chemokine receptor 4 pathway

    PubMed Central

    ZHOU, HAO; YANG, JUNJIE; XIN, TING; ZHANG, TAO; HU, SHUNYIN; ZHOU, SHANSHAN; CHEN, GUANGHUI; CHEN, YUNDAI

    2015-01-01

    Adipose-derived stem cells (ADSCs) are considered a suitable source of cells for the repair of tissue following acute myocardial infarction (AMI); however, the transplantation efficiency of ADSCs remains low. Therefore, identification of an efficient method to enhance the migration of engrafted cells to the target site is required. The present study used exendin-4 (Ex-4), a glucagon-like peptide-1 receptor agonist, to optimize the migratory capacity of ADSCs. The aim was to determine the effect and mechanisms of Ex-4 on the migration of ADSCs to neonatal rat ventricular cardiomyocyte-derived conditioned medium (NRVC-CM). The ADSCs and cardiomyocytes were cultured in vitro. Following incubation of the ADSCs with Ex-4, cell proliferation was measured using an MTT assay and the expression levels of CXC chemokine receptor 4 (CXCR4) were investigated by reverse transctiption quantitative polymerase chain reaction (RT-qPCR), western blot analysis and flow cytometry. In addition, the expression levels of stromal cell-derived factor-1α (SDF-1α) were evaluated in the NRVC-CM treated with Ex-4 by ELISA, RT-qPCR and western blot analysis. The migration of the ADSCs to the NRVC-CM was examined using a Transwell assay. Changes in the protein expression levels of phosphorylated (p−)Akt were examined in the two types of cell by western blot analysis. The results suggested that Ex-4 promoted the proliferation and expression of CXCR4 in the ADSCs, increased the secretion of SDF-1α in the cardiomyocytes and increased the expression levels of p-Akt in both cells. However, the alterations to the SDF-1α/C XC R4 cascade in the cells were abrogated following pretreatment with LY-294002, a phosphoinositide 3-kinase(PI3K) inhibitor. Furthermore, a Transwell migration assay revealed marked translocation of the ADSCs through the membranes, towards the NRVC-CM, following treatment with Ex-4. However, these effects were reduced significantly by pretreatment of the cells with the SDF-1α/CXCR4 cascade antagonist, AMD3100, and the PI3K inhibitor, LY-294002. These results indicated that Ex-4 augmented the SDF-1α/CXCR4 cascade by activating the PI3K/Akt pathways in the ADSCs and NRVCs. Furthermore, enhancement of the PI3K/Akt-SDF-1α/CXCR4 pathway may be important in the migratory response of ADSCs to NRVC-CM in vitro. PMID:25625935

  5. The Effect of Secretory Factors of Adipose-Derived Stem Cells on Human Keratinocytes

    PubMed Central

    Moon, Kyoung Mi; Park, Ye-Hyoung; Lee, Jae Seol; Chae, Yong-Byung; Kim, Moon-Moo; Kim, Dong-Soo; Kim, Byung-Woo; Nam, Soo-Wan; Lee, Jong-Hwan

    2012-01-01

    The beneficial effects of adipose-derived stem cell conditioned medium (ADSC-CM) on skin regeneration have been reported. Although the mechanism of how ADSC-CM promotes skin regeneration is unclear, ADSC-CM contained various growth factors and it is an excellent raw material for skin treatment. ADSC-CM produced in a hypoxia condition of ADSC—in other words, Advanced Adipose-Derived Stem cell Protein Extract (AAPE)—has great merits for skin regeneration. In this study, human primary keratinocytes (HKs), which play fundamental roles in skin tissue, was used to examine how AAPE affects HK. HK proliferation was significantly higher in the experimental group (1.22 μg/mL) than in the control group. DNA gene chip demonstrated that AAPE in keratinocytes (p < 0.05) notably affected expression of 290 identified transcripts, which were associated with cell proliferation, cycle and migration. More keratinocyte wound healing and migration was shown in the experimental group (1.22 μg/mL). AAPE treatment significantly stimulated stress fiber formation, which was linked to the RhoA-ROCK pathway. We identified 48 protein spots in 2-D gel analysis and selected proteins were divided into 64% collagen components and 30% non-collagen components as shown by the MALDI-TOF analysis. Antibody array results contained growth factor/cytokine such as HGF, FGF-1, G-CSF, GM-CSF, IL-6, VEGF, and TGF-β3 differing from that shown by 2-D analysis. Conclusion: AAPE activates HK proliferation and migration. These results highlight the potential of the topical application of AAPE in the treatment of skin regeneration. PMID:22312315

  6. Potential therapeutic effect of the secretome from human uterine cervical stem cells against both cancer and stromal cells compared with adipose tissue stem cells

    PubMed Central

    Seoane, Samuel; Bermúdez, María A.; Lamelas, Maria Luz; Garcia-Caballero, Tomás; Schneider, José; Perez-Fernandez, Roman; Vizoso, Francisco J.

    2014-01-01

    Evidences indicate that tumor development and progression towards a malignant phenotype depend not only on cancer cells themselves, but are also deeply influenced by tumor stroma reactivity. The present study uses mesenchymal stem cells from normal human uterine cervix (hUCESCs), isolated by the minimally invasive method of routine Pap cervical smear, to study their effect on the three main cell types in a tumor: cancer cells, fibroblasts and macrophages. Administration of hUCESCs-conditioned medium (CM) to a highly invasive breast cancer MDA-MB-231 cell line and to human breast tumors with high cell proliferation rates had the effect of reducing cell proliferation, modifying the cell cycle, inducing apoptosis, and decreasing invasion. In a xenograft mouse tumor model, hUCESCs-CM reduced tumor growth and increased overall survival. In cancer-associated fibroblasts, administration of hUCESCs-CM resulted in reduced cell proliferation, greater apoptosis and decreased invasion. In addition, hUCESCs-CM inhibited and reverted macrophage differentiation. The analysis of hUCESCs-CM (fresh and lyophilized) suggests that a complex paracrine signaling network could be implicated in the anti-tumor potential of hUCESCs. In light of their anti-tumor potential, the easy cell isolation method, and the fact that lyophilization of their CM conserves original properties make hUCESCs good candidates for experimental or clinical applications in anticancer therapy. PMID:25296979

  7. Influence of humidity on the initial emittable concentration of formaldehyde and hexaldehyde in building materials: experimental observation and correlation

    PubMed Central

    Huang, Shaodan; Xiong, Jianyin; Cai, Chaorui; Xu, Wei; Zhang, Yinping

    2016-01-01

    Humidity is one of the main environmental factors affecting the emission rate and key parameters of formaldehyde and volatile organic compounds (VOCs) from building materials. Meanwhile, the initial emittable concentration (Cm,0) is proved to be the most sensitive key parameter to the emission behaviours. However, there is no report on the relationship between humidity and Cm,0. In this paper, Cm,0 of formaldehyde and hexaldehyde from a type of medium density fiberboard in absolute humidity (AH) range of 4.6–19.6 g/m3 at 25 °C were tested by virtue of a C-history method. Experimental results indicate that Cm,0 is dramatically dependent on AH, increased by 10 and 2 times for formaldehyde and hexaldehyde when AH rising from 4.6 g/m3 to 19.6 g/m3. A linear relationship between the logarithm of Cm,0 and AH is obtained based on the measured results. In addition, a correlation characterizing the association of emission rate and AH is derived. The effectiveness of the correlation is verified with our experimental results as well as data from literature. With the correlations, the Cm,0 or emission rate different from the test AH conditions can be conveniently obtained. This study should be useful for predicting the emission characteristics of humidity changing scenarios and for source control. PMID:27025353

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

    Misu, Masayasu; Ouji, Yukiteru, E-mail: oujix@naramed-u.ac.jp; Kawai, Norikazu

    In spite of the strong expression of Wnt-10b in melanomas, its role in melanoma cells has not been elucidated. In the present study, the biological effects of Wnt-10b on murine B16F10 (B16) melanoma cells were investigated using conditioned medium from Wnt-10b-producing COS cells (Wnt-CM). After 2 days of culture in the presence of Wnt-CM, proliferation of B16 melanoma cells was inhibited, whereas tyrosinase activity was increased. An in vitro wound healing assay demonstrated that migration of melanoma cells to the wound area was inhibited with the addition of Wnt-CM. Furthermore, evaluation of cellular senescence revealed prominent induction of SA-β-gal-positive senescent cellsmore » in cultures with Wnt-CM. Finally, the growth of B16 melanoma cell aggregates in collagen 3D-gel cultures was markedly suppressed in the presence of Wnt-CM. These results suggest that Wnt-10b represses tumor cell properties, such as proliferation and migration of B16 melanoma cells, driving them toward a more differentiated state along a melanocyte lineage. - Highlights: • Wnt-10b inhibited proliferation and migration of melanoma cells. • Wnt-10b induced tyrosinase activity and senescence of melanoma cells. • Wnt-10b suppressed growth of cell aggregates in collagen 3D-gel cultures. • Wnt-10b represses tumor cell properties, driving them toward a more differentiated state along a melanocyte lineage.« less

  9. Loss of proliferation and differentiation capacity of aged human periodontal ligament stem cells and rejuvenation by exposure to the young extrinsic environment.

    PubMed

    Zheng, Wei; Wang, Shi; Ma, Dandan; Tang, Liang; Duan, Yinzhong; Jin, Yan

    2009-09-01

    The application of periodontal ligament stem cells (PDLSCs) may be effective for periodontal regenerative therapy. As tissue regenerative potential may be negatively regulated by aging, whether aging and its microenvironment modify human PDLSCs remains a question. In this study, we compared the proliferation and differentiation capacity of PDLSCs obtained from young and aged donors. Then, we exposed aged PDLSCs to young periodontal ligament cell-conditioned medium (PLC-CM), and young PDLSCs were exposed to aged PLC-CM. Morphological appearance, colony-forming assay, cell cycle analysis, osteogenic and adipogenic induction media, gene expression of cementoblast phenotype, and in vivo differentiation capacities of PDLSCs were evaluated. PDLSCs obtained from aged donors exhibited decreased proliferation and differentiation capacity when compared with those from young donors. Young PLC-CM enhanced the proliferation and differentiation capacity of PDLSCs from aged donors. Aged PDLSCs induced by young PLC-CM showed enhanced tissue-regenerative capacity to produce cementum/periodontal ligament-like structures, whereas young PDLSCs induced by aged PLC-CM transplants mainly formed connective tissues. To our knowledge, this is the first study to mimic the developmental microenvironment of PDLSCs in vitro, and our data suggest that age influences the proliferation and differentiation potential of human PDLSCs, and that the activity of human PDLSCs can be modulated by the extrinsic microenvironment.

  10. SERPINE2 is a possible candidate promotor for lymph node metastasis in testicular cancer

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

    Nagahara, Akira; Nakayama, Masashi; Oka, Daizo

    2010-01-22

    Testicular germ cell tumors (TGCTs) commonly metastasize to the lymph node or lung. However, it remains unclear which genes are associated with TGCT metastasis. The aim of this study was to identify gene(s) that promoted human TGCT metastasis. We intraperitoneally administered conditioned medium (CM) from JKT-1, a cell-line from a human testicular seminoma, or JKT-HM, a JKT-1 cell sub-line with high metastatic potential, into mice with JKT-1 xenografts. Administration of CM from JKT-HM significantly promoted lymph node metastasis. A cDNA microarray analysis showed that JKT-HM cells highly expressed the Serpine peptidase inhibitor, clade E, member 2 (SERPINE2), which encodes amore » secreted protein. Administration of CM from SERPINE2-silenced JKT-HM cells inhibited lymph node metastasis in the xenograft model, compared with administration of CM from JKT-HM cells. There was no significant difference in xenograft volume. Moreover, administration of CM from SERPINE2-over-expressing JKT-1 was likely to promote lymph node metastasis in the xenograft model. There was no difference in the in vitro proliferation or migration of JKT-1 cells cultured with CM from JKT-HM cells, compared to that with CM from JKT-1. There was no promotion of proliferation or lymphangiogenesis in the xenografts, as measured by Ki-67 and LYVE-1 immunohistochemistry, respectively. Although we could not clarify how SERPINE2 promoted lymph node metastasis, it may be a promoter in the development of lymph node metastasis in the human seminoma cells in a mouse xenograft model.« less

  11. TGF-β signaling is required for maintenance of retinal ganglion cell differentiation and survival.

    PubMed

    Walshe, T E; Leach, L L; D'Amore, P A

    2011-08-25

    To determine the role of TGF-β1 in the maintenance of retinal ganglion cell line (RGC-5) differentiation and integrity. RGC-5 cells were differentiated in media conditioned by human non-pigmented ciliary epithelial cells (HNPE) for 4 days before treatment with TGF-β1 for 24 h. Cells were examined for morphological changes and harvested for western blot and real-time PCR analysis. For study of apoptosis, differentiated RGC-5 cells were grown in serum-free medium for 24 h in the presence or absence of TGF-β1 and collected for Annexin V/Propidium iodide FACs analysis. The role of MAPK pathways in TGF-β1-dependent signaling was determined by treatment with specific inhibitors of ERK, JNK and p38. Differentiation of RGC-5 cells in HNPE-conditioned media (CM) increased the neural cell markers, Brn-3c, NF-160, Thy1.2, Tau and PGP9.5. Treatment with TGF-β1 significantly increased the length of neurites extended by differentiated RGC-5s, concomitant with increased expression of NF-160 and PGP9.5, but not Brn-3c, Thy1.2 or Tau. TGF-β1 also decreased RGC-5 cell apoptosis in serum-free medium. p38 phosphorylation, but not smad2/3, JNK or ERK phosphorylation, was increased in TGF-β1 treated cells. Specific inhibition of p38 signaling reversed TGF-β1 induced neurite growth. These findings demonstrate the induction of RGC-5 cell differentiation by HNPE-derived CM and illustrate a role for TGF-β1 in maintaining RGC-5 cell survival and promoting neurite outgrowth through p38 MAPK. Copyright © 2011 IBRO. Published by Elsevier Ltd. All rights reserved.

  12. ORION’S VEIL: MAGNETIC FIELD STRENGTHS AND OTHER PROPERTIES OF A PDR IN FRONT OF THE TRAPEZIUM CLUSTER

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

    Troland, T. H.; Goss, W. M.; Brogan, C. L.

    2016-07-01

    We present an analysis of physical conditions in the Orion Veil, an atomic photon-dominated region (PDR) that lies just in front (≈2 pc) of the Trapezium stars of Orion. This region offers an unusual opportunity to study the properties of PDRs, including the magnetic field. We have obtained 21 cm H i and 18 cm (1665 and 1667 MHz) OH Zeeman effect data that yield images of the line-of-sight magnetic field strength B {sub los} in atomic and molecular regions of the Veil. We find B {sub los} ≈ −50 to −75 μ G in the atomic gas across muchmore » of the Veil (25″ resolution) and B {sub los} ≈ −350 μ G at one position in the molecular gas (40″ resolution). The Veil has two principal H i velocity components. Magnetic and kinematical data suggest a close connection between these components. They may represent gas on either side of a shock wave preceding a weak-D ionization front. Magnetic fields in the Veil H i components are 3–5 times stronger than they are elsewhere in the interstellar medium where N (H) and n (H) are comparable. The H i components are magnetically subcritical (magnetically dominated), like the cold neutral medium, although they are about 1 dex denser. Comparatively strong fields in the Veil H i components may have resulted from low-turbulence conditions in the diffuse gas that gave rise to OMC-1. Strong fields may also be related to magnetostatic equilibrium that has developed in the Veil since star formation. We also consider the location of the Orion-S molecular core, proposing a location behind the main Orion H{sup +} region.« less

  13. Large-volume iodinated contrast medium extravasation: low frequency and good outcome after conservative management in a single-centre cohort of more than 67,000 patients.

    PubMed

    Ko, Chih-Hsiang; Tay, Shee Yen; Chang, Hsiu-Chin; Chan, Wing P

    2018-06-12

    Our aim was to retrospectively investigate the frequency and outcome of large-volume iodinated contrast medium (CM) extravasation in our institution and to compare our management protocol to current practice. Institutional review board approval was obtained, and informed consent was waived because the study was retrospective. From January 2008 to September 2016, radiological examinations with intravenous non-ionic iodinated CM administration were performed in 67,129 patients. Contrast medium extravasation events on CT scans and intravenous pyelograms but not on angiograms were included. All data were collected prospectively and stratified according to age, injection method (manual vs auto-injection), prevention of extravasation by various means (including intercom alarm), management of extravasation (routine application of silver sulfadiazine ointment, clobetasol propionate cream, and damp gauze at room temperature), etc. RESULTS: The incidence of large-volume CM extravasation was very low (0.04% [27/67,129] overall; 0.03% related to manual injection [age range, 59-92 years; mean, 75.4 years], and 0.045% related to auto-injection [age range, 36-86 years; mean, 65.8 years]). The CM extravasation volume in majority of patients was 20-40 ml in 5 of 9 patients (55.6%) in the manual injection group and 14 of 18 (77.8%) in the auto-injection group. Swelling and pain were the most common symptoms. No patient developed severe signs or needed surgical intervention. Results show a very low incidence of large-volume CM extravasation without severe complications or sequelae. The casual effect between our protocols and good outcome cannot be scrutinised thoroughly because the study lacks a control group and is retrospective. • The incidence of large-volume contrast medium extravasation (≥20 ml) was 0.04%. • No patient needed surgical intervention, and most recovered within 7 days. • Each element of our management protocol contributed to good outcome.

  14. Fluid flow releases fibroblast growth factor-2 from human aortic smooth muscle cells

    NASA Technical Reports Server (NTRS)

    Rhoads, D. N.; Eskin, S. G.; McIntire, L. V.

    2000-01-01

    This study tested the hypothesis that fluid shear stress regulates the release of fibroblast growth factor (FGF)-2 from human aortic smooth muscle cells. FGF-2 is a potent mitogen that is involved in the response to vascular injury and is expressed in a wide variety of cell types. FGF-2 is found in the cytoplasm of cells and outside cells, where it associates with extracellular proteoglycans. To test the hypothesis that shear stress regulates FGF-2 release, cells were exposed to flow, and FGF-2 amounts were measured from the conditioned medium, pericellular fraction (extracted by heparin treatment), and cell lysate. Results from the present study show that after 15 minutes of shear stress at 25 dyne/cm(2) in a parallel-plate flow system, a small but significant fraction (17%) of the total FGF-2 was released from human aortic smooth muscle cells. FGF-2 levels in the circulating medium increased 10-fold over medium from static controls (P<0.01). A 50% increase in FGF-2 content versus control (P<0.01) was found in the pericellular fraction (extracted by heparin treatment). Furthermore, a significant decrease in FGF-2 was detected in the cell lysate, indicating that FGF-2 was released from inside the cell. Cell permeability studies with fluorescent dextran were performed to examine whether transient membrane disruption caused FGF-2 release. Flow cytometry detected a 50% increase in mean fluorescence of cells exposed to 25 dyne/cm(2) versus control cells. This indicates that the observed FGF-2 release from human aortic smooth muscle cells is likely due to transient membrane disruption on initiation of flow.

  15. Ultrafine particle emission characteristics of diesel engine by on-board and test bench measurement.

    PubMed

    Huang, Cheng; Lou, Diming; Hu, Zhiyuan; Tan, Piqiang; Yao, Di; Hu, Wei; Li, Peng; Ren, Jin; Chen, Changhong

    2012-01-01

    This study investigated the emission characteristics of ultrafine particles based on test bench and on-board measurements. The bench test results showed the ultrafine particle number concentration of the diesel engine to be in the range of (0.56-8.35) x 10(8) cm(-3). The on-board measurement results illustrated that the ultrafine particles were strongly correlated with changes in real-world driving cycles. The particle number concentration was down to 2.0 x 10(6) cm(-3) and 2.7 x 10(7) cm(-3) under decelerating and idling operations and as high as 5.0 x 10(8) cm(-3) under accelerating operation. It was also indicated that the particle number measured by the two methods increased with the growth of engine load at each engine speed in both cases. The particle number presented a "U" shaped distribution with changing speed at high engine load conditions, which implies that the particle number will reach its lowest level at medium engine speeds. The particle sizes of both measurements showed single mode distributions. The peak of particle size was located at about 50-80 nm in the accumulation mode particle range. Nucleation mode particles will significantly increase at low engine load operations like idling and decelerating caused by the high concentration of unburned organic compounds.

  16. Local application of periodontal ligament stromal cells promotes soft tissue regeneration.

    PubMed

    Baik, H S; Park, J; Lee, K J; Chung, C

    2014-09-01

    To test the potential stimulatory effect of local application of periodontal ligament (PDL) stromal cells on soft tissue regeneration. Fluorescently labeled PDL cells outgrown from extracted human premolars or phosphate-buffered saline were locally injected to the cutaneous wounds created on mice. Soft tissue regeneration was evaluated for 14 days using photographs and histomorphometry. PDL cell engraftment was tracked with confocal microscopy. To detect the paracrine effect of the PDL cells on soft tissue regeneration, PDL cell-conditioned medium (CM) was evaluated for the concentration of secretory factors, transforming growth factor-beta 1 (TGFβ1). The effect of PDL CM on the proliferation and migration of dermal fibroblast and keratinocyte was tested using MTT assay and migration assay. The application of PDL cells significantly promoted soft tissue regeneration compared with the application of PBS. Self-replicating PDL cells were engrafted into the hair follicles of the host tissue. Dermal fibroblast proliferation and keratinocyte migration were significantly enhanced by the treatment with PDL CM. Physiologically significant amount of TGFβ1 was secreted from PDL cells into the CM. Local injection of PDL cells promoted soft tissue regeneration in part by the enhancement of fibroblast proliferation and keratinocyte migration through a paracrine mechanism. © 2013 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

  17. Air sparging: Air-water mass transfer coefficients

    NASA Astrophysics Data System (ADS)

    Braida, Washington J.; Ong, Say Kee

    1998-12-01

    Experiments investigating the mass transfer of several dissolved volatile organic compounds (VOCs) across the air-water interface were conducted using a single-air- channel air-sparging system. Three different porous media were used in the study. Air velocities ranged from 0.2 cm s-1 to 2.5 cm s-1. The tortuosity factor for each porous medium and the air-water mass transfer coefficients were estimated by fitting experimental data to a one-dimensional diffusion model. The estimated mass transfer coefficients KG ranged from 1.79 × 10-3 cm min-1 to 3.85 × 10-2 cm min-1. The estimated lumped gas phase mass transfer coefficients KGa were found to be directly related to the air diffusivity of the VOC, air velocity, and particle size, and inversely related to the Henry's law constant of the VOCs. Of the four parameters investigated, the parameter that controlled or had a dominant effect on the lumped gas phase mass transfer coefficient was the air diffusivity of the VOC. Two empirical models were developed by correlating the Damkohler and the modified air phase Sherwood numbers with the air phase Peclet number, Henry's law constant, and the reduced mean particle size of porous media. The correlation developed in this study may be used to obtain better predictions of mass transfer fluxes for field conditions.

  18. Mesenchymal stem cell's secretome promotes selective enrichment of cancer stem-like cells with specific cytogenetic profile.

    PubMed

    Jiménez, Gema; Hackenberg, Michael; Catalina, Purificación; Boulaiz, Houria; Griñán-Lisón, Carmen; García, María Ángel; Perán, Macarena; López-Ruiz, Elena; Ramírez, Alberto; Morata-Tarifa, Cynthia; Carrasco, Esther; Aguilera, Margarita; Marchal, Juan Antonio

    2018-08-10

    Cancer stem cells (CSCs) are responsible for tumor initiation, metastasis and cancer recurrence, however the involvement of microenvironment is crucial. Here, we have analyzed how human mesenchymal stem cells (MSCs)-derived conditioned medium (CM) affect colon and melanoma CSCs enrichment and maintenance. Our results strongly suggest that the secretome of CM-MSCs selects and maintains subpopulations with high expression of CSCs markers and ALDH1 activity, low proliferation rates with G1 phase arrest, and notably retain in vivo these properties. Cytogenetic analyses indicated that CM-cultured cells contain alterations in chromosome 17 (17q25). Subsequent SKY-FISH analyses suggested that genes located in 17q25 might be involved in stem-cell maintenance. The characterization of secreted proteins present in CM-MSCs revealed that four cytokines and seven growth factors are directly linked to the CSCs enrichment reported in this study. Further analyses revealed that the combination of just IL6 and HGF is enough to provide cancer cells with better stemness properties. In conclusion, this study demonstrates how specific chromosomal alterations present in CSCs subpopulations might represent an advantage for their in vitro maintenance and in vivo stemness properties. Copyright © 2018 Elsevier B.V. All rights reserved.

  19. Factors Associated with Performance-based Physical Function of Older Veterans of the PLAAF: A Pilot Study

    PubMed Central

    Chen, Da-Wei; Jin, Yan-Bin; Liu, Wei; Du, Wen-Jin; Li, Hua-Jun; Chen, Jin-Wen; Xu, Wei

    2014-01-01

    [Purpose] This study investigated the factors associated with performance-based physical function of older veterans of the People’s Liberation Army Air Force of China (PLAAF). [Subjects and Methods] A cross-sectional survey of 146 older veterans of the PLAAF was carried out. Their physical function was measured using the Chinese Mini-Physical Performance Testing (CM-PPT). The demographics and health status (including physical measures, blood chemical tests, chronic diseases, and number of morbidities) were collected from health examination reports and computer records of case history. Cognition was measured using the Mini-Mental Status Examination (MMSE). [Results] In multiple linear regressions, age, MMSE, Parkinsonism, and chronic obstructive pulmonary disease were independently associated with CM-PPT, while previous stroke and albumin level reached borderline statistical significance. The association between the number of morbidities and CM-PPT was significant after adjustment for MMSE and demographics. The CM-PPT of low (0 or 1), medium (2 to 4) and high count (5 or more) morbidities were 11.3±3.9, 10.2±4.1, 6.1±3.8 respectively, and the difference among these three groups was significant. [Conclusion] Some modified conditions and the number of chronic diseases might be associated with the physical function of older veterans of the PLAAF. PMID:24648625

  20. Holocene sedimentation in the shallow nearshore zone off Nauset Inlet, Cape Cod, Massachusetts

    USGS Publications Warehouse

    Aubrey, D.G.; Twichell, D.C.; Pfirman, S.L.

    1982-01-01

    Present conditions and sedimentary evolution of the shallow offshore region near Nauset Inlet on Cape Cod, Massachusetts were clarified using high-resolution seismic-reflection profiles, sidescan-sonar records, surface grab samples and current meter measurements. The study area contains three provinces: (1) a nearshore province (shallower than 18 m) with a relatively steep slope (0.6°) and a cover of medium sand; (2) a northern offshore province covered with coarse sand, gravel, and boulders, interpreted to be glacial drift; and (3) a southern offshore province with a gentle seaward-dipping slope (0.3°) and a surface sediment of coarse sand. The glacial drift exposed in the northern offshore province can be traced southward under the coarse sand province. The overlying fill is comprised of either outwash sediment derived from the Pleistocene South Channel ice lobe to the east or Holocene-aged marine sediments eroded from seacliffs to the north. Latest Holocene sediment appears to be limited to the zone shoreward of 18 m where the medium sand occurs.Near-bottom mean flows (measured over two winter months in 10 m water depth) average 6 cm sec−1 to the south. Mean flows exceeded 20 cm sec−1approx. 23% of the time. Ninety percent of the flows exceeding 20 cm sec−1were directed to the south, reflecting the dominant atmospheric forcing during these winter months. Waves had an average variance of 650 cm2 with variance exceeding 5000 cm2, 3% of the time, indicating moderate wave activity.Present processes are actively reshaping the nearshore province, which is characterized by many east to northeast-trending shore-oblique channels that do not extend seaward of the 18-m contour. Coarse sand in the floors of these channels suggests they may be erosional features, and the presence of megaripples oriented perpendicular to the channel axes indicates active transport in these channels. Megaripple orientation and the current and wave regime of the study area support a rip-current origin for these channels.

  1. Modulation of chondrogenic differentiation of human mesenchymal stem cells in jellyfish collagen scaffolds by cell density and culture medium.

    PubMed

    Pustlauk, W; Paul, B; Brueggemeier, S; Gelinsky, M; Bernhardt, A

    2017-06-01

    Studies on tissue-engineering approaches for the regeneration of traumatized cartilage focus increasingly on multipotent human mesenchymal stem cells (hMSCs) as an alternative to autologous chondrocytes. The present study applied porous scaffolds made of collagen from the jellyfish Rhopilema esculentum for the in vitro chondrogenic differentiation of hMSCs. Culture conditions in those scaffolds differ from conditions in high-density pellet cultures, making a re-examination of these data necessary. We systematically investigated the influence of seeding density, basic culture media [Dulbecco's modified Eagle's medium (DMEM), α-minimum essential medium (α-MEM)] with varying glucose content and supplementation with fetal calf serum (FCS) or bovine serum albumin (BSA) on the chondrogenic differentiation of hMSCs. Gene expression analyses of selected markers for chondrogenic differentiation and hypertrophic development were conducted. Furthermore, the production of cartilage extracellular matrix (ECM) was analysed by quantification of sulphated glycosaminoglycan and collagen type II contents. The strongest upregulation of chondrogenic markers, along with the highest ECM deposition was observed in scaffolds seeded with 2.4 × 10 6 cells/cm 3 after cultivation in high-glucose DMEM and 0.125% BSA. Lower seeding densities compared to high-density pellet cultures were sufficient to induce in vitro chondrogenic differentiation of hMSCs in collagen scaffolds, which reduces the amount of cells required for the seeding of scaffolds and thus the monolayer expansion period. Furthermore, examination of the impact of FCS and α-MEM on chondrogenic MSC differentiation is an important prerequisite for the development of an osteochondral medium for simultaneous osteogenic and chondrogenic differentiation in biphasic scaffolds for osteochondral tissue regeneration. Copyright © 2015 John Wiley & Sons, Ltd. Copyright © 2015 John Wiley & Sons, Ltd.

  2. Investment appraisal of automatic milking and conventional milking technologies in a pasture-based dairy system.

    PubMed

    Shortall, J; Shalloo, L; Foley, C; Sleator, R D; O'Brien, B

    2016-09-01

    The successful integration of automatic milking (AM) systems and grazing has resulted in AM becoming a feasible alternative to conventional milking (CM) in pasture-based systems. The objective of this study was to identify the profitability of AM in a pasture-based system, relative to CM herringbone parlors with 2 different levels of automation, across 2 farm sizes, over a 10-yr period following initial investment. The scenarios which were evaluated were (1) a medium farm milking 70 cows twice daily, with 1 AM unit, a 12-unit CM medium-specification (MS) parlor and a 12-unit CM high-specification (HS) parlor, and (2) a large farm milking 140 cows twice daily with 2 AM units, a 20-unit CM MS parlor and a 20-unit CM HS parlor. A stochastic whole-farm budgetary simulation model combined capital investment costs and annual labor and maintenance costs for each investment scenario, with each scenario evaluated using multiple financial metrics, such as annual net profit, annual net cash flow, total discounted net profitability, total discounted net cash flow, and return on investment. The capital required for each investment was financed from borrowings at an interest rate of 5% and repaid over 10-yr, whereas milking equipment and building infrastructure were depreciated over 10 and 20 yr, respectively. A supporting labor audit (conducted on both AM and CM farms) showed a 36% reduction in labor demand associated with AM. However, despite this reduction in labor, MS CM technologies consistently achieved greater profitability, irrespective of farm size. The AM system achieved intermediate profitability at medium farm size; it was 0.5% less profitable than HS technology at the large farm size. The difference in profitability was greatest in the years after the initial investment. This study indicated that although milking with AM was less profitable than MS technologies, it was competitive when compared with a CM parlor of similar technology. Copyright © 2016 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

  3. Effect of cow manure and empty fruit bunches application treated with different fertilizers on growth and yield of chili (Capsicum annum)

    NASA Astrophysics Data System (ADS)

    Ghazali, Mohd Rashdan; Mutalib, Sahilah Abd.; Abdullah, Aminah

    2016-11-01

    Study on the comparison of cow manure (CM) and empty fruit bunches (EFB) compost application as planting medium was conducted using four different treatments of fertilizer (without fertilizer, chemical fertilizer, organic fertilizer, and both fertilizer) on growth and yield of chili (Capsicum annum). The experiment started on August until December 2014 which consisted of eight treatments and were laid in a completely randomized block design (CRBD) with three replications. Variety chili that was used was Cilibangi 3. The seed was planted inside the tray for one week and transferred into the polybag containing growth media consisted of soil, compost (CM or EFB compost) and sand with ratio 3:2:1. Treatments without fertilizer were acted as a control. Throughout the study, plant growth performance and yield were recorded. The highest height of the plants for CM compost was 100.8 cm using chemical fertilizer and have significant different between the groups. For EFB compost was 92.7 cm using also chemical fertilizer but no significant different between the groups. The highest fruits weight per plant for CM compost was 485.67 g treated with both fertilizers and for EFB compost was 420.17 g treated with chemical fertilizer. Analysis of variance (ANOVA) table stated that fruits weight per plant has significant different for both planting medium with the fertilizer treatment. For the highest total fruits per plant, CM compost recorded about average 55 fruits per plant using both fertilizers and EFB compost recorded around 45 fruit per plant using chemical fertilizer. There was significantly different for total fruits per plant for both planting medium with the fertilizer treatment according to the ANOVA table. For CM, the ripening time was around 102-112 days and for EFB compost was around 96-110 days. Thus, application of CM compost treated with both chemical and organic fertilizers demonstrated better growth and fruit yield. While EFB compost was better growth and fruit yield when treated with chemical fertilizer.

  4. Combination of the Auxins NAA, IBA, and IAA with GA3 Improves the Commercial Seed-Tuber Production of Potato (Solanum tuberosum L.) under In Vitro Conditions

    PubMed Central

    Kumlay, Ahmet Metin

    2014-01-01

    The study compared the effects of 1.0 × MS medium containing various concentrations of α-naphthaleneacetic acid (NAA), indole-3-acetic acid (IAA), and indole-3-butyric acid (IBA), alone or in combination with gibberellic acid (GA3) in micropropagation of three potato (Solanum tuberosum L.) cultivars Pasinler, Granola, and Caspar using binodal stem cuttings. The results testified improved regeneration on 1.0 × MS medium containing variants of NAA, IAA, and IBA plus GA3 on all cultivars. The minimum days to shoot induction on three cultivars ranged 4.25–5 d on 1.0 × MS medium containing 0.25 mg L−1  GA3 + 1 mg L−1 NAA. The longest shoots (11.8 cm), maximum number of nodes (13.50), and maximum number of leaves (11.00) were recorded on cv. Caspar on 1.0 × MS medium containing 1 mg L−1  NAA + 0.25 mg L−1 GA3. The minimum time to root induction (12.25 d) was noted on cv. Pasinler on the same medium. All of the regenerated shoots could be easily rooted. The results showed that the combined effect of various concentrations of NAA, IAA, and IBA plus GA3 was more pronounced compared to the auxins used alone. The results of this research are of significant importance for potato breeders. PMID:25028654

  5. Combination of the auxins NAA, IBA, and IAA with GA3 improves the commercial seed-tuber production of potato (Solanum tuberosum L.) under in vitro conditions.

    PubMed

    Kumlay, Ahmet Metin

    2014-01-01

    The study compared the effects of 1.0 × MS medium containing various concentrations of α-naphthaleneacetic acid (NAA), indole-3-acetic acid (IAA), and indole-3-butyric acid (IBA), alone or in combination with gibberellic acid (GA3) in micropropagation of three potato (Solanum tuberosum L.) cultivars Pasinler, Granola, and Caspar using binodal stem cuttings. The results testified improved regeneration on 1.0 × MS medium containing variants of NAA, IAA, and IBA plus GA3 on all cultivars. The minimum days to shoot induction on three cultivars ranged 4.25-5 d on 1.0 × MS medium containing 0.25 mg L(-1) GA3 + 1 mg L(-1) NAA. The longest shoots (11.8 cm), maximum number of nodes (13.50), and maximum number of leaves (11.00) were recorded on cv. Caspar on 1.0 × MS medium containing 1 mg L(-1) NAA + 0.25 mg L(-1) GA3. The minimum time to root induction (12.25 d) was noted on cv. Pasinler on the same medium. All of the regenerated shoots could be easily rooted. The results showed that the combined effect of various concentrations of NAA, IAA, and IBA plus GA3 was more pronounced compared to the auxins used alone. The results of this research are of significant importance for potato breeders.

  6. The Redshifted Hydrogen Balmer and Metastable He 1 Absorption Line System in Mini-FeLoBAL Quasar SDSS J112526.12+002901.3: A Parsec-scale Accretion Inflow?

    NASA Astrophysics Data System (ADS)

    Shi, Xi-Heng; Jiang, Peng; Wang, Hui-Yuan; Zhang, Shao-Hua; Ji, Tuo; Liu, Wen-Juan; Zhou, Hong-Yan

    2016-10-01

    The accretion of the interstellar medium onto central super-massive black holes is widely accepted as the source of the gigantic energy released by the active galactic nuclei. However, few pieces of observational evidence have been confirmed directly demonstrating the existence of the inflows. The absorption line system in the spectra of quasar SDSS J112526.12+002901.3 presents an interesting example in which the rarely detected hydrogen Balmer and metastable He I absorption lines are found redshifted to the quasar's rest frame along with the low-ionization metal absorption lines Mg II, Fe II, etc. The repeated SDSS spectroscopic observations suggest a transverse velocity smaller than the radial velocity. The motion of the absorbing medium is thus dominated by infall. The He I* lines present a powerful probe to the strength of ionizing flux, while the Balmer lines imply a dense environment. With the help of photoionization simulations, we find that the absorbing medium is exposed to the radiation with ionization parameter U ≈ 10-1.8, and the density is n({{H}})≈ {10}9 {{cm}}-3. Thus the absorbing medium is located ˜4 pc away from the central engine. According to the similarity in the distance and physical conditions between the absorbing medium and the torus, we strongly propose the absorption line system as a candidate for the accretion inflow, which originates in the inner surface of the torus.

  7. Defining the role of mesenchymal stromal cells on the regulation of matrix metalloproteinases in skeletal muscle cells

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

    Sassoli, Chiara; Nosi, Daniele; Tani, Alessia

    Recent studies indicate that mesenchymal stromal cell (MSC) transplantation improves healing of injured and diseased skeletal muscle, although the mechanisms of benefit are poorly understood. In the present study, we investigated whether MSCs and/or their trophic factors were able to regulate matrix metalloproteinase (MMP) expression and activity in different cells of the muscle tissue. MSCs in co-culture with C2C12 cells or their conditioned medium (MSC-CM) up-regulated MMP-2 and MMP-9 expression and function in the myoblastic cells; these effects were concomitant with the down-regulation of the tissue inhibitor of metalloproteinases (TIMP)-1 and -2 and with increased cell motility. In the singlemore » muscle fiber experiments, MSC-CM administration increased MMP-2/9 expression in Pax-7{sup +} satellite cells and stimulated their mobilization, differentiation and fusion. The anti-fibrotic properties of MSC-CM involved also the regulation of MMPs by skeletal fibroblasts and the inhibition of their differentiation into myofibroblasts. The treatment with SB-3CT, a potent MMP inhibitor, prevented in these cells, the decrease of α-smooth actin and type-I collagen expression induced by MSC-CM, suggesting that MSC-CM could attenuate the fibrogenic response through mechanisms mediated by MMPs. Our results indicate that growth factors and cytokines released by these cells may modulate the fibrotic response and improve the endogenous mechanisms of muscle repair/regeneration. - Highlights: • MSC-CM contains paracrine factors that up-regulate MMP expression and function in different skeletal muscle cells. • MSC-CM promotes myoblast and satellite cell migration, proliferation and differentiation. • MSC-CM negatively interferes with fibroblast-myoblast transition in primary skeletal fibroblasts. • Paracrine factors from MSCs modulate the fibrotic response and improve the endogenous mechanisms of muscle regeneration.« less

  8. Factors secreted from dental pulp stem cells show multifaceted benefits for treating acute lung injury in mice.

    PubMed

    Wakayama, Hirotaka; Hashimoto, Naozumi; Matsushita, Yoshihiro; Matsubara, Kohki; Yamamoto, Noriyuki; Hasegawa, Yoshinori; Ueda, Minoru; Yamamoto, Akihito

    2015-08-01

    Acute respiratory distress syndrome (ARDS) is a severe inflammatory disorder characterized by acute respiratory failure, resulting from severe, destructive lung inflammation and irreversible lung fibrosis. We evaluated the use of stem cells derived from human exfoliated deciduous teeth (SHEDs) or SHED-derived serum-free conditioned medium (SHED-CM) as treatments for bleomycin (BLM)-induced mice acute lung injury (ALI), exhibiting several pathogenic features associated with the human disease ARDS. Mice with BLM-induced ALI with or without SHED or SHED-CM treatment were examined for weight loss and survival. The lung tissue was characterized by histological and real-time quantitative polymerase chain reaction analysis. The effects of SHED-CM on macrophage differentiation in vitro were also assessed. A single intravenous administration of either SHEDs or SHED-CM attenuated the lung injury and weight loss in BLM-treated mice and improved their survival rate. Similar recovery levels were seen in the SHEDs and SHED-CM treatment groups, suggesting that SHED improves ALI by paracrine mechanisms. SHED-CM contained multiple therapeutic factors involved in lung-regenerative mechanisms. Importantly, SHED-CM attenuated the BLM-induced pro-inflammatory response and generated an anti-inflammatory/tissue-regenerating environment, accompanied by the induction of anti-inflammatory M2-like lung macrophages. Furthermore, SHED-CM promoted the in vitro differentiation of bone marrow-derived macrophages into M2-like cells, which expressed high levels of Arginase1, CD206 and Ym-1. Our results suggest that SHED-secreted factors provide multifaceted therapeutic effects, including a strong M2-inducing activity, for treating BLM-induced ALI. This work may open new avenues for research on stem cell-based ARDS therapies. Copyright © 2015 International Society for Cellular Therapy. Published by Elsevier Inc. All rights reserved.

  9. Neuroprotective Potential of Mesenchymal Stem Cell-Based Therapy in Acute Stages of TNBS-Induced Colitis in Guinea-Pigs

    PubMed Central

    Robinson, Ainsley M.; Miller, Sarah; Payne, Natalie; Boyd, Richard; Sakkal, Samy; Nurgali, Kulmira

    2015-01-01

    Background & Aims The therapeutic benefits of mesenchymal stem cells (MSCs), such as homing ability, multipotent differentiation capacity and secretion of soluble bioactive factors which exert neuroprotective, anti-inflammatory and immunomodulatory properties, have been attributed to attenuation of autoimmune, inflammatory and neurodegenerative disorders. In this study, we aimed to determine the earliest time point at which locally administered MSC-based therapies avert enteric neuronal loss and damage associated with intestinal inflammation in the guinea-pig model of colitis. Methods At 3 hours after induction of colitis by 2,4,6-trinitrobenzene-sulfonate (TNBS), guinea-pigs received either human bone marrow-derived MSCs, conditioned medium (CM), or unconditioned medium by enema into the colon. Colon tissues were collected 6, 24 and 72 hours after administration of TNBS. Effects on body weight, gross morphological damage, immune cell infiltration and myenteric neurons were evaluated. RT-PCR, flow cytometry and antibody array kit were used to identify neurotrophic and neuroprotective factors released by MSCs. Results MSC and CM treatments prevented body weight loss, reduced infiltration of leukocytes into the colon wall and the myenteric plexus, facilitated repair of damaged tissue and nerve fibers, averted myenteric neuronal loss, as well as changes in neuronal subpopulations. The neuroprotective effects of MSC and CM treatments were observed as early as 24 hours after induction of inflammation even though the inflammatory reaction at the level of the myenteric ganglia had not completely subsided. Substantial number of neurotrophic and neuroprotective factors released by MSCs was identified in their secretome. Conclusion MSC-based therapies applied at the acute stages of TNBS-induced colitis start exerting their neuroprotective effects towards enteric neurons by 24 hours post treatment. The neuroprotective efficacy of MSC-based therapies can be exerted independently to their anti-inflammatory effects. PMID:26397368

  10. Regenerative and reparative effects of human chorion-derived stem cell conditioned medium on photo-aged epidermal cells

    PubMed Central

    Li, Qiankun; Chen, Yan; Ma, Kui; Zhao, Along; Zhang, Cuiping; Fu, Xiaobing

    2016-01-01

    ABSTRACT Epidermal cells are an important regenerative source for skin wound healing. Aged epidermal cells have a low ability to renew themselves and repair skin injury. Ultraviolet (UV) radiation, particularly UVB, can cause photo-aging of the skin by suppressing the viability of human epidermal cells. A chorion-derived stem cell conditioned medium (CDSC-CNM) is thought to have regenerative properties. This study aimed to determine the regenerative effects of CDSC-CNM on UVB-induced photo-aged epidermal cells. Epidermal cells were passaged four times and irradiated with quantitative UVB, and non-irradiated cells served as a control group. Cells were then treated with different concentrations of CDSC-CNM. Compared to the non-irradiated group, the proliferation rates and migration rates of UVB-induced photo-aged epidermal cells significantly decreased (p < 0.05) with increasing intracellular radical oxygen species (ROS) generation and DNA damage. After treatment with CDSC-CNM, photo-aged epidermal cells significantly improved their viability, and their ROS generation and DNA damage decreased. The secretory factors in CDSC-CNM, including epidermal growth factor (EGF), transforming growth factor-β (TGF-β), interleukin (IL)-6, and IL-8 and the related signaling pathway protein levels, increased compared to the control medium (CM). The potential regenerative and reparative effects of CDSC-CNM indicate that it may be a candidate material for the treatment of prematurely aged skin. The functions of the secretory factors and the mechanisms of CDSC-CNM therapy deserve further attention. PMID:27097375

  11. Effects of crop load on distribution and utilization of 13C and 15N and fruit quality for dwarf apple trees.

    PubMed

    Ding, Ning; Chen, Qian; Zhu, Zhanling; Peng, Ling; Ge, Shunfeng; Jiang, Yuanmao

    2017-10-26

    In order to define the effects of fruit crop load on the distribution and utilization of carbon and nitrogen in dwarf apple trees, we conducted three crop load levels (High-crop load, 6 fruits per trunk cross-sectional area (cm 2 , TCA)), Medium-crop load (4 fruits cm -2 TCA), Low-crop load (2 fruits cm -2 TCA)) in 2014 and 2015. The results indicated that the 15 N derived from fertilizer (Ndff) values of fruits decreased with the reduction of crop load, but the Ndff values of annual branches, leaves and roots increased. The plant 15 N-urea utilization rates on Medium and Low-crop load were 1.12-1.35 times higher than the High-crop load. With the reduction of crop load, the distribution rate of 13 C and 15 N in fruits was gradually reduced, but in contrast, the distribution of 13 C and 15 N gradually increased in annual branches, leaves and roots. Compared with High-crop load, the Medium and Low-crop load significantly improved fruit quality p < 0.05. Hence, controlling fruit load effectively regulated the distribution of carbon and nitrogen in plants, improved the nitrogen utilization rate and fruit quality. The appropriate crop load level for mature M.26 interstocks apple orchards was deemed to be 4.0 fruits cm -2 TCA.

  12. Spectrophotometric determination of trace carbaryl in water and grain samples by inhibition of the rhodamine-B oxidation.

    PubMed

    Gupta, Nirja; Pillai, Ajai Kumar; Parmar, Prachi

    2015-03-15

    A novel, sensitive, selective and simple kinetic spectrophotometric method has been developed for determination of trace levels of carbaryl based on its inhibitory effect on the oxidation of rhodamine-B by chlorine and bromine released from reaction of potassium bromate with hydrochloric acid in micellar medium. A linear relationship was observed between the inhibitory effect and the concentration of the compound. The absorbance was monitored at the maximum wavelength of 555 nm. The effect of different parameters such as pH, temperature and concentration of rhodamine-B, potassium bromate and surfactant on the reaction were investigated and optimum conditions were established. Under the selected experimental conditions, carbaryl was determined in the range of 0.04-0.4 μg mL(-1). Sandell's sensitivity and molar absorptivity were found to be 0.00055 μg cm(-2) and 3.658×10(5) L mol(-1) cm(-1) respectively. The proposed method was applied satisfactorily for the determination of carbaryl in water and different grain samples. The results were compared with those obtained by reference method and were found to be in agreement. Copyright © 2014 Elsevier B.V. All rights reserved.

  13. Affective valence, stimulus attributes, and P300: color vs. black/white and normal vs. scrambled images.

    PubMed

    Cano, Maya E; Class, Quetzal A; Polich, John

    2009-01-01

    Pictures from the International Affective Picture System (IAPS) were selected to manipulate affective valence (unpleasant, neutral, pleasant) while keeping arousal level the same. The pictures were presented in an oddball paradigm, with a visual pattern used as the standard stimulus. Subjects pressed a button whenever a target was detected. Experiment 1 presented normal pictures in color and black/white. Control stimuli were constructed for both the color and black/white conditions by randomly rearranging 1 cm square fragments of each original picture to produce a "scrambled" image. Experiment 2 presented the same normal color pictures with large, medium, and small scrambled condition (2, 1, and 0.5 cm squares). The P300 event-related brain potential demonstrated larger amplitudes over frontal areas for positive compared to negative or neutral images for normal color pictures in both experiments. Attenuated and nonsignificant valence effects were obtained for black/white images. Scrambled stimuli in each study yielded no valence effects but demonstrated typical P300 topography that increased from frontal to parietal areas. The findings suggest that P300 amplitude is sensitive to affective picture valence in the absence of stimulus arousal differences, and that stimulus color contributes to ERP valence effects.

  14. In Vitro Impact of Conditioned Medium From Demineralized Freeze-Dried Bone on Human Umbilical Endothelial Cells.

    PubMed

    Harnik, Branko; Miron, Richard J; Buser, Daniel; Gruber, Reinhard

    2017-03-01

    Angiogenesis is essential for the consolidation of bone allografts. The underlying molecular mechanism, however, remains unclear. Soluble factors released from demineralized freeze-dried bone target mesenchymal cells; however, their effect on endothelial cells has not been investigated so far. The aim of the present study was therefore to examine the effect of conditioned medium from demineralized freeze-dried bone on human umbilical endothelial cells in vitro. Conditioned medium was first prepared from demineralized freeze-dried bone following 24 hours incubation at room temperature to produce demineralized bone conditioned media. Thereafter, conditioned medium was used to stimulate human umbilical vein endothelial cells in vitro by determining the cell response based on viability, proliferation, expression of apoptotic genes, a Boyden chamber to determine cell migration, and the formation of branches. The authors report here that conditioned medium decreased viability and proliferation of endothelial cells. Neither of the apoptotic marker genes was significantly altered when endothelial cells were exposed to conditioned medium. The Boyden chamber revealed that endothelial cells migrate toward conditioned medium. Moreover, conditioned medium moderately stimulated the formation of branches. These findings support the concept that conditioned medium from demineralized freeze-dried bone targets endothelial cells by decreasing their proliferation and enhancing their motility under these in vitro conditions.

  15. WISDOM GPR performance assessment in a cold artificial environment

    NASA Astrophysics Data System (ADS)

    Dechambre, M.; Ciarletti, V.; Biancheri-Astier, M.; Saintenoy, A.; Costard, F.; Hassen-Khodja, R.

    2012-04-01

    The WISDOM (Water Ice Subsurface Deposit Observation on Mars) GPR is one of the instruments that have been selected as part of the Pasteur payload of ESA's 2018 ExoMars Rover mission. WISDOM has been designed to obtain information about the nature of the subsurface along the rover path with the objective to explore the first ~ 3 m of the soil with a vertical resolution of a few centimetres. The sub-surface properties that can be addressed with WISDOM are variations in composition, texture, stratification (e.g., number, thickness and orientation of layers), the presence of unconformities and other structural characteristics (such as fractures and the deformation of strata). It is then essential to quantify the performances of WISDOM in controlled conditions, and several full polarimetric measurements have been carried out with the prototype in a cold artificial environment. The main objectives are the detection of different interface between homogeneous materials with WISDOM. The characterization of the material (porosity, % of water, dielectric properties, thickness and depth, temperature ...) is well-controlled. The cold room facility of IDES at Orsay (France) has been used, the ambient temperature ranged from -7° C to -10° C. A tank laying on the metallic floor (height: 0.5m, width: 0.80 m, length: 1.20m) in macrolon can contain liquid or frozen water or layers (dielectric contrasts) of home-maid permafrost (frozen saturated sand) with and without embedded objects or fractures. The temperature inside the medium (ice or permafrost) is controlled, the radar antennas are put on a sheet of polystyrene over the tank. Frequent measurements were performed (every 2cm) along a track from one side to the other side of the tank. The experimental conditions were: (1)dry cold sand (Fontainebleau sand) : porosity 35% density 2,67 (2) saturated wet sand : 35% of water (3) permafrost (frozen saturated sand) : 35% of ice content 1 layer: 3 consecutive experiments : 10cm dry sand ( 1) 10cm saturated sand (2) 10cm permafrost(3) 2 layers :previous 10cm permafrost in the bottom +3 consecutive experiments : 10cm dry sand ( 1) 10cm saturated sand (2) 10cm permafrost(3) . Basalt rocks and air fractures are or are not embedded in the layers Values of the permittivity of dry sand and permafrost were retrieve by two different ways. 1.Retrieval of the sand and permafrost permittivity from delay measurements knowing the layer thickness d ɛr = ct/2d = n2 2. Retrieval of the sand and permafrost permittivity from amplitude measurements knowing a calibration reference (reflection over a metallic plate), R is the Fresnel coefficient between the air and the medium. Aint/ = R = n- 1-,ɛ = 1+-R-= n2 Aair n +1 r 1- R Results : Internal layering is observed. The transition between dry sand and permafrost can be detected. The permittivity can be retrieved from delay or amplitude measurements as well from delay measurements : dry sand ɛr = 2.71 permafrost ɛr = 3.72 from amplitude measurements : dry sand ɛr = 2.73 permafrost ɛr = 3.35 Embedded objects are detected, Fracture and its orientation is detected.

  16. Adipocyte differentiation influences the proliferation and migration of normal and tumoral breast epithelial cells.

    PubMed

    Creydt, Virginia Pistone; Sacca, Paula Alejandra; Tesone, Amelia Julieta; Vidal, Luciano; Calvo, Juan Carlos

    2010-01-01

    Stromal tissue regulates the development and differentiation of breast epithelial cells, with adipocytes being the main stromal cell type. The aim of the present study was to evaluate the effect of adipocyte differentiation on proliferation and migration, as well as to assess the activity of heparanase and metalloproteinase-9 (MMP-9), in normal (NMuMG) and tumoral (LM3) murine breast epithelial cells. NMuMG and LM3 cells were grown on irradiated 3T3-L1 cells (stromal support, SS) at various degrees of differentiation [preadipocytes (preA), poorly differentiated adipocytes (pDA) and mature adipocytes (MA)] and/or were incubated in the presence of conditioned medium (CM) derived from each of these three types of differentiated cells. Cells grown on a plastic support or in fresh medium served as the controls. Cell proliferation was measured with a commercial colorimetric kit, and the motility of the epithelial cells was evaluated by means of a wound-healing assay. Heparanase activity was assessed by quantifying heparin degradation, and the expression of MMP-9 was determined using Western blotting. The results indicate that cell proliferation was increased after 24 and 48 h in the NMuMG and LM3 cells grown on preA, pDA and MA SS. In the NMuMG cells cultured on SS in the presence of all three types of CM, proliferation was enhanced. LM3 cell migration was increased in the presence of all three types of CM and in cells grown on preA SS. Heparanase activity was increased in the NMuMG cells incubated with all three types of CM, and in the LM3 cells incubated with the CM from pDA and MA. Both the NMuMG and LM3 cell lines presented basal expression of MMP-9; however, a significant increase in MMP-9 expression was observed in the LM3 cells incubated with each of the three types of CM. In conclusion, adipocyte differentiation influences normal and tumoral breast epithelial cell proliferation and migration. Heparanase and MMP-9 appear to be involved in this regulation. The experimental model presented in this study is in keeping with the characteristics of the physiological environment of breast epithelial cells, in terms of both the soluble and insoluble factors present and the stromal structure per se.

  17. Measurements of the global 21-cm signal from the Cosmic Dawn

    NASA Astrophysics Data System (ADS)

    Bernardi, Gianni

    2018-05-01

    The sky-averaged (global) 21-cm signal is a very promising probe of the Cosmic Dawn, when the first luminous sources were formed and started to shine in a substantially neutral intergalactic medium. I here report on the status and early result of the Large-Aperture Experiment to Detect the Dark Age that focuses on observations of the global 21-cm signal in the 16 <~ z <~ 30 range.

  18. The endothelial cell secretome as a novel treatment to prime adipose-derived stem cells for improved wound healing in diabetes.

    PubMed

    Fromer, Marc W; Chang, Shaohua; Hagaman, Ashleigh L R; Koko, Kiavash R; Nolan, Ryan S; Zhang, Ping; Brown, Spencer A; Carpenter, Jeffrey P; Caputo, Francis J

    2017-07-28

    Chronic wounds are a common surgical problem exacerbated by diabetes and ischemia. Although adipose-derived stem cells (ASCs) have shown promise as a wound healing therapy, their function and proliferation are hindered in diabetes. This study examines the ability of the human umbilical vein endothelial cell (HUVEC) secretome to reverse the deleterious effects of high glucose concentrations on ASCs through priming, thereby enhancing their ability to participate in angiogenesis and wound healing. Institutional review board-approved human ASCs were cultured in M199 medium with or without glucose (30 mmol/L). HUVEC were grown in 30 mmol/L glucose-containing M199 medium; the resulting conditioned medium (HUVEC-CM) was collected every 3 days and used to prime ASCs. An aliquot of HUVEC-CM was heated (85°C for 30 minutes) to produce thermal denaturation of protein. Viability, proliferation, and endothelial differentiation were measured by MTT assays, growth curves, and quantitative polymerase chain reaction, respectively. A Matrigel assay was used to assess the ability of primed ASCs to participate in capillary-like tube formation. An Institutional Animal Care and Use Committee-approved in vivo murine model of diabetic and ischemic hindlimbs was used to evaluate the angiogenic potential of primed stem cells. Human ASCs were cultured with either control M199 or HUVEC-CM. Mice were randomized to a control group, an unprimed ASC group, or a HUVEC-primed ASC group. Cellular therapies were injected into the ischemic muscle. Thirty days later, slides were made. Microvessels were counted by three blinded observers. MTT assays revealed that HUVEC-priming induced a 1.5 times increase in cell viability over diabetic controls. This promoting effect was lost with heated HUVEC-CM (P < .001), indicating that the active molecules are of protein origin. After 9 days, ASCs cultured in 30 mmol/L glucose solution showed a 14% reduction in growth from nondiabetic controls (P = .013) and exhibited atrophic morphology. Conversely, diabetic HUVEC-primed stem cells demonstrated a nearly four-fold increase in proliferation (P < .05) and took on a fusiform, endothelial-like phenotype. Polymerase chain reaction demonstrated enhanced expression of CD31 messenger RNA by 4.7-fold after 14 days in the HUVEC-primed group, and endothelial nitric oxide synthase messenger RNA messenger RNA was increased 20.1-fold from controls. Unlike unprimed controls, HUVEC-primed ASCs readily formed capillary-like tube networks on Matrigel. Diabetic mice that were injected with HUVEC-primed ASCs demonstrated greater vessel density than both controls (2.1-fold) and unprimed stem cell treatments (P < .001). HUVECs secrete protein factors that significantly increase proliferation and endothelial differentiation of ASCs under diabetic conditions. Injection of ischemic hindlimbs in diabetic mice with HUVEC-primed ASCs leads to enhanced angiogenesis. Copyright © 2017 Society for Vascular Surgery. Published by Elsevier Inc. All rights reserved.

  19. Laboratory and on-road evaluations of cabin air filters using number and surface area concentration monitors.

    PubMed

    Qi, Chaolong; Stanley, Nick; Pui, David Y H; Kuehn, Thomas H

    2008-06-01

    An automotive cabin air filter's effectiveness for removing airborne particles was determined both in a laboratory wind tunnel and in vehicle on-road tests. The most penetrating particle size for the test filter was approximately 350 nm, where the filtration efficiency was 22.9 and 17.4% at medium and high fan speeds, respectively. The filtration efficiency increased for smaller particles and was 43.9% for 100 nm and 72.0% for 20 nm particles at a medium fan speed. We determined the reduction in passenger exposure to particles while driving in freeway traffic caused by a vehicle ventilation system with a cabin air filter installed. Both particle number and surface area concentration measurements were made inside the cabin and in the surrounding air. At medium fan speed, the number and surface area concentration-based exposure reductions were 65.6 +/- 6.0% and 60.6 +/- 9.4%, respectively. To distinguish the exposure reduction contribution from the filter alone and the remainder of the ventilation system, we also performed tests with and without the filter in place using the surface area monitors. The ventilation system operating in the recirculation mode with the cabin air filter installed provided the maximum protection, reducing the cabin particle concentration exponentially over time and usually taking only 3 min to reach 10 microm2/cm3 (a typical office air condition) under medium fan speed.

  20. An interplanetary magnetic field ensemble at 1 AU

    NASA Technical Reports Server (NTRS)

    Matthaeus, W. H.; Goldstein, M. L.; King, J. H.

    1985-01-01

    A method for calculation ensemble averages from magnetic field data is described. A data set comprising approximately 16 months of nearly continuous ISEE-3 magnetic field data is used in this study. Individual subintervals of this data, ranging from 15 hours to 15.6 days comprise the ensemble. The sole condition for including each subinterval in the averages is the degree to which it represents a weakly time-stationary process. Averages obtained by this method are appropriate for a turbulence description of the interplanetary medium. The ensemble average correlation length obtained from all subintervals is found to be 4.9 x 10 to the 11th cm. The average value of the variances of the magnetic field components are in the approximate ratio 8:9:10, where the third component is the local mean field direction. The correlation lengths and variances are found to have a systematic variation with subinterval duration, reflecting the important role of low-frequency fluctuations in the interplanetary medium.

  1. Beam-beam interaction study of medium energy eRHIC

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

    Hao,Y.; Litvinenko, V. N.; Ptitsyn, V.

    Medium Energy eRHIC (MeRHIC), the first stage design of eRHIC, includes a multi-pass ERL that provides 4GeV high quality electron beam to collide with the ion beam of RHIC. It delivers a minimum luminosity of 10{sup 32} cm{sup -2}s{sup -1}. Beam-beam effects present one of major factors limiting the luminosity of colliders. In this paper, both beam-beam effects on the electron beam and the proton beam in MeRHIC are investigated. The beam-beam interaction can induce a head-tail type instability of the proton beam referred to as the kink instability. Thus, beam stability conditions should be established to avoid proton beammore » loss. Also, the electron beam transverse disruption by collisions has to be evaluated to ensure that the beam quality is good enough for the energy recovery pass. The relation of proton beam stability, electron disruption and consequential luminosity are carried out after thorough discussion.« less

  2. Effect of pulse electric fields (PEF) on accumulation of magnesium and zinc ions in Saccharomyces cerevisiae cells.

    PubMed

    Pankiewicz, Urszula; Sujka, Monika; Włodarczyk-Stasiak, Marzena; Mazurek, Artur; Jamroz, Jerzy

    2014-08-15

    Cultures of Saccharomyces cerevisiae were treated with PEF to improve simultaneous accumulation of magnesium and zinc ions in the biomass. The results showed that the ions concentration in the medium and their mutual interactions affect accumulation in cells. Increasing the concentration of one ion in the medium reduced the accumulation of the second one, in the control as well as in the cells treated with PEF. Under optimized conditions, that is, on 15 min exposure of the 20 h grown culture to PEF of 5.0 kV/cm and 20 μs pulse width, accumulation of magnesium and zinc in yeast biomass reached maximum levels of 2.85 and 11.41 mg/gd.m., respectively, To summarize, optimization of ion pair concentration and PEF parameters caused a 1.5 or 2-fold increase of magnesium and zinc accumulation, respectively, in S. cerevisiae. Copyright © 2014 Elsevier Ltd. All rights reserved.

  3. Multi-level multi-thermal-electron FDTD simulation of plasmonic interaction with semiconducting gain media: applications to plasmonic amplifiers and nano-lasers.

    PubMed

    Chen, X; Bhola, B; Huang, Y; Ho, S T

    2010-08-02

    Interactions between a semiconducting gain medium and confined plasmon-polaritons are studied using a multilevel multi-thermal-electron finite-difference time-domain (MLMTE-FDTD) simulator. We investigated the amplification of wave propagating in a plasmonic metal-semiconductor-metal (MSM) waveguide filled with semiconductor gain medium and obtained the conditions required to achieve net optical gain. The MSM gain waveguide is used to form a plasmonic semiconductor nano-ring laser(PSNRL) with an effective mode volume of 0.0071 microm3, which is about an order of magnitude smaller than the smallest demonstrated integrated photonic crystal based laser cavities. The simulation shows a lasing threshold current density of 1kA/cm2 for a 300 nm outer diameter ring cavity with 80 nm-wide ring. This current density can be realistically achieved in typical III-V semiconductor, which shows the experimental feasibility of the proposed PSNRL structure.

  4. Physiological considerations in applying laboratory-determined buoyant densities to predictions of bacterial and protozoan transport in groundwater: Results of in-situ and laboratory tests

    USGS Publications Warehouse

    Harvey, R.W.; Metge, D.W.; Kinner, N.; Mayberry, N.

    1997-01-01

    Buoyant densities were determined for groundwater bacteria and microflagellates (protozoa) from a sandy aquifer (Cape Cod, MA) using two methods: (1) density-gradient centrifugation (DGC) and (2) Stoke's law approximations using sedimentation rates observed during natural-gradient injection and recovery tests. The dwarf (average cell size, 0.3 ??m), unattached bacteria inhabiting a pristine zone just beneath the water table and a majority (~80%) of the morphologically diverse community of free- living bacteria inhabiting a 5-km-long plume of organically-contaminated groundwater had DGC-determined buoyant densities <1.019 g/cm3 before culturing. In the aquifer, sinking rates for the uncultured 2-??m size class of contaminant plume bacteria were comparable to that of the bromide tracer (1.9 x 10-3 M), also suggesting a low buoyant density. Culturing groundwater bacteria resulted in larger (0.8-1.3 ??m), less neutrally- buoyant (1.043-1.081 g/cm3) cells with potential sedimentation rates up to 64-fold higher than those predicted for the uncultured populations. Although sedimentation generally could be neglected in predicting subsurface transport for the community of free-living groundwater bacteria, it appeared to be important for the cultured isolates, at least until they readapt to aquifer conditions. Culturing-induced alterations in size of the contaminant-plume microflagellates (2-3 ??m) were ameliorated by using a lower nutrient, acidic (pH 5) porous growth medium. Buoyant densities of the cultured microflagellates were low, i.e., 1.024-1.034 g/cm3 (using the DGC assay) and 1.017-1.039 g/cm3 (estimated from in-situ sedimentation rates), suggesting good potential for subsurface transport under favorable conditions.

  5. Loss of the endothelial glycocalyx is associated with increased E-selectin mediated adhesion of lung tumour cells to the brain microvascular endothelium.

    PubMed

    Rai, Srijana; Nejadhamzeeigilani, Zaynab; Gutowski, Nicholas J; Whatmore, Jacqueline L

    2015-09-25

    Arrest of metastasising lung cancer cells to the brain microvasculature maybe mediated by interactions between ligands on circulating tumour cells and endothelial E-selectin adhesion molecules; a process likely to be regulated by the endothelial glycocalyx. Using human cerebral microvascular endothelial cells and non-small cell lung cancer (NSCLC) cell lines, we describe how factors secreted by NSCLC cells i.e. cystatin C, cathepsin L, insulin-like growth factor-binding protein 7 (IGFBP7), vascular endothelial growth factor (VEGF) and tumour necrosis factor-alpha (TNF-α), damage the glycocalyx and enhance initial contacts between lung tumour and cerebral endothelial cells. Endothelial cells were treated with tumour secreted-proteins or lung tumour conditioned medium (CM). Surface levels of E-selectin were quantified by ELISA. Adhesion of A549 and SK-MES-1 cells was examined under flow conditions (1 dyne/cm(2)). Alterations in the endothelial glycocalyx were quantified by binding of fluorescein isothiocyanate-linked wheat germ agglutinin (WGA-FITC). A549 and SK-MES-1 CM and secreted-proteins significantly enhanced endothelial surface E-selectin levels after 30 min and 4 h and tumour cell adhesion after 30 min, 4 and 24 h. Both coincided with significant glycocalyx degradation; A549 and SK-MES-1 CM removing 55 ± 12 % and 58 ± 18.7 % of WGA-FITC binding, respectively. Inhibition of E-selectin binding by monoclonal anti-E-selectin antibody completely attenuated tumour cell adhesion. These data suggest that metastasising lung cancer cells facilitate their own adhesion to the brain endothelium by secreting factors that damage the endothelial glycocalyx, resulting in exposure of the previously shielded adhesion molecules and engagement of the E-selectin-mediated adhesion axis.

  6. Cardiac Stem Cell Secretome Protects Cardiomyocytes from Hypoxic Injury Partly via Monocyte Chemotactic Protein-1-Dependent Mechanism

    PubMed Central

    Park, Chi-Yeon; Choi, Seung-Cheol; Kim, Jong-Ho; Choi, Ji-Hyun; Joo, Hyung Joon; Hong, Soon Jun; Lim, Do-Sun

    2016-01-01

    Cardiac stem cells (CSCs) were known to secrete diverse paracrine factors leading to functional improvement and beneficial left ventricular remodeling via activation of the endogenous pro-survival signaling pathway. However, little is known about the paracrine factors secreted by CSCs and their roles in cardiomyocyte survival during hypoxic condition mimicking the post-myocardial infarction environment. We established Sca-1+/CD31− human telomerase reverse transcriptase-immortalized CSCs (Sca-1+/CD31− CSCshTERT), evaluated their stem cell properties, and paracrine potential in cardiomyocyte survival during hypoxia-induced injury. Sca-1+/CD31− CSCshTERT sustained proliferation ability even after long-term culture exceeding 100 population doublings, and represented multi-differentiation potential into cardiomyogenic, endothelial, adipogenic, and osteogenic lineages. Dominant factors secreted from Sca-1+/CD31− CSCshTERT were EGF, TGF-β1, IGF-1, IGF-2, MCP-1, HGF R, and IL-6. Among these, MCP-1 was the most predominant factor in Sca-1+/CD31− CSCshTERT conditioned medium (CM). Sca-1+/CD31− CSCshTERT CM increased survival and reduced apoptosis of HL-1 cardiomyocytes during hypoxic injury. MCP-1 silencing in Sca-1+/CD31− CSCshTERT CM resulted in a significant reduction in cardiomyocyte apoptosis. We demonstrated that Sca-1+/CD31− CSCshTERT exhibited long-term proliferation capacity and multi-differentiation potential. Sca-1+/CD31− CSCshTERT CM protected cardiomyocytes from hypoxic injury partly via MCP-1-dependent mechanism. Thus, they are valuable sources for in vitro and in vivo studies in the cardiovascular field. PMID:27231894

  7. Interleukin-1beta may mediate insulin resistance in liver-derived cells in response to adipocyte inflammation.

    PubMed

    Nov, Ori; Kohl, Ayelet; Lewis, Eli C; Bashan, Nava; Dvir, Irit; Ben-Shlomo, Shani; Fishman, Sigal; Wueest, Stephan; Konrad, Daniel; Rudich, Assaf

    2010-09-01

    Central obesity is frequently associated with adipose tissue inflammation and hepatic insulin resistance. To identify potential individual mediators in this process, we used in vitro systems and assessed if insulin resistance in liver cells could be induced by secreted products from adipocytes preexposed to an inflammatory stimulus. Conditioned medium from 3T3-L1 adipocytes pretreated without (CM) or with TNFalpha (CM-TNFalpha) was used to treat Fao hepatoma cells. ELISAs were used to assess the concentration of several inflammatory mediators in CM-TNFalpha. CM-TNFalpha-treated Fao cells exhibited about 45% diminution in insulin-stimulated phosphorylation of insulin receptor, insulin receptor substrate proteins, protein kinase B, and glycogen synthase kinase-3 as compared with CM-treated cells, without changes in the total abundance of these protein. Insulin increased glycogenesis by 2-fold in CM-treated Fao cells but not in cells exposed to CM-TNFalpha. Expression of IL-1beta mRNA was elevated 3-fold in TNFalpha-treated adipocytes, and CM-TNFalpha had 10-fold higher concentrations of IL-1beta but not TNFalpha or IL-1alpha. IL-1beta directly induced insulin resistance in Fao, HepG2, and in primary rat hepatocytes. Moreover, when TNFalpha-induced secretion/production of IL-1beta from adipocytes was inhibited by the IL-1 converting enzyme (ICE-1) inhibitor II (Ac-YVAD-CMK), insulin resistance was prevented. Furthermore, liver-derived cells treated with IL-1 receptor antagonist were protected against insulin resistance induced by CM-TNFalpha. Finally, IL-1beta secretion from human omental fat explants correlated with body mass index (R(2) = 0.639, P < 0.01), and the resulting CM induced insulin resistance in HepG2 cells, inhibitable by IL-1 receptor antagonist. Our results suggest that adipocyte-derived IL-1beta may constitute a mediator in the perturbed cross talk between adipocytes and liver cells in response to adipose tissue inflammation.

  8. Bauhinia variegata L.

    Treesearch

    K.F Connor

    2002-01-01

    Bauhinia variegata is a small to medium-sized evergreen or deciduous tree that reaches 1.8 to 7.6 m in height and up to 20.3 cm in diameter. The species grows well on soils of medium fertility that are either droughty or moist; it is not tolerate of nutrient-poor sites. Although reproduction is abundant, B. variegata spreads...

  9. The Effect of Aggressive Corrosion Mediums on the Microstructure and Properties of Mild Steel

    NASA Astrophysics Data System (ADS)

    Araoyinbo, A. O.; Salleh, M. A. A. Mohd; Rahmat, A.; Azmi, A. I.; Rahim, W. M. F. Wan Abd; Achitei, D. C.; Jin, T. S.

    2018-06-01

    Mild steel is known to be one of the major construction materials and have been extensively used in most chemical and material industries due to its interesting properties which can be easily altered to suit various application areas. In this research, mild steel is exposed to different aggressive mediums in order to observe the effect of these interactions on its surface morphology and properties. The mild steel used was cut into dimensions of 7 cm length and width of 3 cm. The aggressive mediums used are 100 mls of aqueous solution of hydrochloric acid, sodium hydroxide (40 g/L), and sodium chloride (35 g/L) at room temperature. The characterizations performed are the hardness test with the Rockwell hardness tester, the surface morphology by optical microscope, surface roughness and the weight loss from the immersion test. It was observed that the hardness value and the weight loss for the different cut samples of mild steel immersed in the different aggressive mediums reduces with prolong exposure and severe pitting form of corrosion was present on its surface.

  10. Fabrication and characterization of fine ceramic based on alumina, bentonite, and glass bead

    NASA Astrophysics Data System (ADS)

    Sebayang, P.; Nurdina; Simbolon, S.; Kurniawan, C.; Yunus, M.; Setiadi, E. A.; Sitorus, Z.

    2018-03-01

    Fabrication of fine ceramics based on alumina, bentonite and glass bead has been carried out by powder metallurgy. The preparation of powder has been performed using High Energy Milling (HEM) with wet milling process and using toluene as medium for 2 hours. The powder milling result was dried in oven at 100 °C for 24 hours. After that, the powder was compacted into pellet by using hydraulic press with 80 kgf/cm2 pressure at room temperature. Then, the pellet was sintered at 900 °C for 4 hours. Materials characterization such as physical properties (true density, bulk density, porosity, and water absorption), average particle diameter, hardness, microstructure and phase were measured by Archimedes method, Particle Size Analyzer (PSA), Hardness Vickers (HV), Scanning Electron Microscope (SEM-EDX) and X-Ray Diffraction (XRD). From the result, the optimum condition is sample D (with addition of 30 wt.% γ-Al2O3) with sintering temperature of 900 °C for 4 hours. At this condition, these properties were measured: average particle diameter of 4.27 μm, true density of 2.32 g/cm3, porosity of 5.57%, water absorption of 2.46%, bulk density of 2.39 g/cm3, and hardness of 632 HV. The fine ceramic has four phases with albite (Al2NaO8Si3) and quartz (SiO2) as dominant phases and corundum (Al2O3) and nepheline (AlNaO4Si) as minor phases.

  11. JAK inhibition alleviates the cellular senescence-associated secretory phenotype and frailty in old age

    PubMed Central

    Xu, Ming; Tchkonia, Tamara; Ding, Husheng; Ogrodnik, Mikolaj; Lubbers, Ellen R.; Pirtskhalava, Tamar; White, Thomas A.; Johnson, Kurt O.; Stout, Michael B.; Mezera, Vojtech; Giorgadze, Nino; Jensen, Michael D.; LeBrasseur, Nathan K.; Kirkland, James L.

    2015-01-01

    Chronic, low grade, sterile inflammation frequently accompanies aging and age-related diseases. Cellular senescence is associated with the production of proinflammatory chemokines, cytokines, and extracellular matrix (ECM) remodeling proteases, which comprise the senescence-associated secretory phenotype (SASP). We found a higher burden of senescent cells in adipose tissue with aging. Senescent human primary preadipocytes as well as human umbilical vein endothelial cells (HUVECs) developed a SASP that could be suppressed by targeting the JAK pathway using RNAi or JAK inhibitors. Conditioned medium (CM) from senescent human preadipocytes induced macrophage migration in vitro and inflammation in healthy adipose tissue and preadipocytes. When the senescent cells from which CM was derived had been treated with JAK inhibitors, the resulting CM was much less proinflammatory. The administration of JAK inhibitor to aged mice for 10 wk alleviated both adipose tissue and systemic inflammation and enhanced physical function. Our findings are consistent with a possible contribution of senescent cells and the SASP to age-related inflammation and frailty. We speculate that SASP inhibition by JAK inhibitors may contribute to alleviating frailty. Targeting the JAK pathway holds promise for treating age-related dysfunction. PMID:26578790

  12. Shewanella frigidimarina microbial fuel cells and the influence of divalent cations on current output.

    PubMed

    Fitzgerald, Lisa A; Petersen, Emily R; Leary, Dagmar H; Nadeau, Lloyd J; Soto, Carissa M; Ray, Richard I; Little, Brenda J; Ringeisen, Bradley R; Johnson, Glenn R; Vora, Gary J; Biffinger, Justin C

    2013-02-15

    The genes involved in the proposed pathway for Shewanella extracellular electron transfer (EET) are highly conserved. While extensive studies involving EET from a fresh water Shewanella microbe (S. oneidensis MR-1) to soluble and insoluble electron acceptors have been published, only a few reports have examined EET from marine strains of Shewanella. Thus, Shewanella frigidimarina (an isolate from Antarctic Sea ice) was used within miniature microbial fuel cells (mini-MFC) to evaluate potential power output. During the course of this study several distinct differences were observed between S. oneidensis MR-1 and S. frigidimarina under comparable conditions. The maximum power density with S. frigidimarina was observed when the anolyte was half-strength marine broth (1/2 MB) (0.28 μW/cm(2)) compared to Luria-Bertani (LB) (0.07 μW/cm(2)) or a defined growth minimal medium (MM) (0.02 μW/cm(2)). The systematic modification of S. frigidimarina cultured in 1/2 MB and LB with divalent cations shows that a maximum current output can be generated independent of internal ionic ohmic losses and the presence of external mediators. Published by Elsevier B.V.

  13. AMP-activated kinase mediates adipose stem cell-stimulated neuritogenesis of PC12 cells.

    PubMed

    Tan, B; Luan, Z; Wei, X; He, Y; Wei, G; Johnstone, B H; Farlow, M; Du, Y

    2011-05-05

    Adipose tissue stroma contains a population of mesenchymal stem cells, which support repair of damaged tissues through the protective effects of secreted trophic factors. Neurotrophic factors, including nerve growth factor (NGF) have been identified in media collected from cultured adipose-derived stem cells (ASC). We previously demonstrated that administration of cell-free ASC conditioned medium (ASC-CM) at 24 h after injury reduced lesion volume and promoted functional recovery in a rat model of neonatal brain hypoxic-ischemic (HI) injury. The timing of administration well after the peak in neural cell apoptosis in the affected region suggests that regeneration of lost neurons is promoted by factors in ASC-CM. In this study, we determined which of the factors in ASC-CM could induce neurogenesis by testing the ability of the mixture, either whole or after inactivating specific components, to stimulate neurite outgrowth in vitro using the neurogenic cell line PC12. Neuritogenesis in PC12 cells treated with ASC-CM was observed at a level comparable to that observed with purified recombinant NGF. It was observed that NGF in ASC-CM was mainly responsible for inducing PC12 cell neuritogenesis. Interestingly, both ASC-CM and NGF induced PC12 cell neuritogenesis through activation of the AMP-activated kinase (AMPK) pathway which is the central protein involved in controlling many critical functions in response to changes in the cellular energy status. Pharmacological and genetic inhibition of AMPK activity greatly reduced neuritogenesis in PC12 cells. These results suggest that, in addition to possessing neuroprotective properties, ASC-CM mediates repair of damaged tissues through inducing neuronal differentiation via NGF-induced AMPK activation. Copyright © 2011 IBRO. Published by Elsevier Ltd. All rights reserved.

  14. In vitro effect of low-level laser on odontoblast-like cells

    NASA Astrophysics Data System (ADS)

    Oliveira, C. F.; Basso, F. G.; Lins, E. C.; Kurachi, C.; Hebling, J.; Bagnato, V. S.; de Souza Costa, C. A.

    2011-02-01

    The aim of this study was to evaluate the metabolism of odontoblast-like MDPC-23 cells subjected to direct LLL irradiation. The cells were seeded (20,000 cells/well) in 24-well plates and incubated for 24 hours at 37°C. After this period, the culture medium (DMEM) was replaced by fresh DMEM supplemented with 2 or 5% (stress induction by nutritional deficit) or 10% fetal bovine serum (FBS). The cells were exposed to laser doses of 2, 4, 10, 15 and 25 J/cm2 from a near infrared InGaAsP diode laser prototype (LASERTable; 780±3 nm, 40 mW). One control group (sham irradiation) was established for each experimental condition (laser dose x FBS supplementation). Three and 72 hours after the last irradiation, cells were analyzed with respect to metabolism, morphology, total protein expression and alkaline phosphatase (ALP) activity. Higher metabolism and total protein expression were observed 72 hours after the last irradiation at the doses of 15 and 25 J/cm2 (Mann-Whitney; p < 0.05). Higher ALP activity was obtained with 5% FBS when the cells were irradiated with doses of 2 and 10 J/cm2. For the dose of 25 J/cm2, the highest ALP activity was observed with 10% FBS. It was concluded that the LLLT parameters used in this study stimulated the metabolic activity of the MDPC-23 cells, especially at the doses of 15 and 25 J/cm2.

  15. Extended Renal Outcomes with Use of Iodixanol versus Iohexol after Coronary Angiography

    PubMed Central

    Horrigan, Mark; Mcintosh, Elizabeth; Bellomo, Rinaldo

    2014-01-01

    The impact of isoosmolar versus low-osmolar contrast media (CM) administration on contrast-induced acute kidney injury (CI-AKI) and extended renal dysfunction (ERD) is unclear. We retrospectively examined incidences of CI-AKI and ERD in patients who received iodixanol (isoosmolar) versus iohexol (low-osmolar) during angiography for cardiac indications. Of 713 patients, 560 (cohort A), 190 (cohort B), and 172 (cohort C) had serum creatinine monitored at 3 days, 30 days, and 6 months after angiography, respectively. 18% of cohort A developed CI-AKI, which was more common with iodixanol than iohexol (22% versus 13%, P = 0.006). However, patients given iodixanol were older with lower baseline estimated glomerular filtration rates (eGFR). On multivariate analysis, independent associations with higher CI-AKI risk include age >65 years, female gender, cardiac failure, ST-elevation myocardial infarction, intra-aortic balloon pump, and critical illness, but not CM type, higher CM load, or eGFR < 45 mL/min/1.73 m2. 32% of cohort B and 34% of cohort C had ERD at 30 days and 6 months, while 44% and 41% of subcohorts had ERD at 90 days and 1 year, respectively. CI-AKI, but not CM type, was associated with medium- and longer-term ERD, with 3-fold higher risk. Advanced age, emergent cardiac conditions, and critical illness are stronger predictors of CI-AKI, compared with CM-related factors. CI-AKI predicts longer-term ERD. PMID:25180184

  16. Persisting effects of armored military maneuvers on some soils of the Mojave Desert

    USGS Publications Warehouse

    Prose, D.V.

    1985-01-01

    Soil compaction and substrate modification produced during large-scale armored military maneuvers in the early 1940s were examined in 1981 at seven sites in California's eastern Mojave Desert Recording penetrometer measurements show that tracks left by a single pass of an M3 "medium" tank have average soil resistance values that are 50% greater than those of the surrounding untracked soil in the upper 20 cm At one site, measurements made along short segments of track that have been visually eliminated by erosion and deposition processes show a 73% increase in penetrometer resistance over adjacent, undisturbed soils Dirt roadways at three former base camp locations could not be penetrated below 5-10 cm because of extreme compaction Soil bulk density was not as sensitive an indicator of soil compaction as was penetrometer resistance Density values in the upper 10 cm of soil are not significantly different between tank tracks and undisturbed soils at most sites, and roadways at two base camps show an average increase in bulk density of only 12% over adjacent soils. Trench excavations across tank tracks show that physical modifications of the substrate can extend vertically beneath a track to a depth of 25 cm and outward from a track's edge to 50 cm These soil disturbances are probably major factors that encourage accelerated soil erosion throughout the manuever area and also retard or prevent the return of vegetation to pre-disturbance conditions ?? 1985 Springer-Verlag New York Inc.

  17. Gastric cancer tissue-derived mesenchymal stem cells impact peripheral blood mononuclear cells via disruption of Treg/Th17 balance to promote gastric cancer progression.

    PubMed

    Wang, Mei; Chen, Bin; Sun, Xiao-Xian; Zhao, Xiang-Dong; Zhao, Yuan-Yuan; Sun, Li; Xu, Chang-Gen; Shen, Bo; Su, Zhao-Liang; Xu, Wen-Rong; Zhu, Wei

    2017-12-01

    Gastric cancer tissue-derived mesenchymal stem cells (GC-MSCs) are important resident stromal cells in the tumor microenvironment (TME) and have been shown to play a key role in gastric cancer progression. Whether GC-MSCs exert a tumor-promoting function by affecting anti-tumor immunity is still unclear. In this study, we used GC-MSC conditioned medium (GC-MSC-CM) to pretreat peripheral blood mononuclear cells (PBMCs) from healthy donors. We found that GC-MSC-CM pretreatment markedly reversed the inhibitory effect of PBMCs on gastric cancer growth in vivo, but did not affect functions of PBMCs on gastric cancer cell proliferation, cell cycle and apoptosis in vitro. PBMCs pretreated with GC-MSC-CM significantly promoted gastric cancer migration and epithelial-mesenchymal transition in vitro and liver metastases in vivo. Flow cytometry analysis showed that GC-MSC-CM pretreatment increased the proportion of Treg cells and reduced that of Th17 cells in PBMCs. CFSE labeling and naïve CD4 + T cells differentiation analysis revealed that GC-MSC-CM disrupted the Treg/Th17 balance in PBMCs by suppressing Th17 cell proliferation and inducing differentiation of Treg cells. Overall, our collective results indicate that GC-MSCs impair the anti-tumor immune response of PBMCs through disruption of Treg/Th17 balance, thus providing new evidence that gastric cancer tissue-derived MSCs contribute to the immunosuppressive TME. Copyright © 2017 Elsevier Inc. All rights reserved.

  18. Further Evaluation of Scaling Methods for Rotorcraft Icing

    NASA Technical Reports Server (NTRS)

    Tsao, Jen-Ching; Kreeger, Richard E.

    2012-01-01

    The paper will present experimental results from two recent icing tests in the NASA Glenn Icing Research Tunnel (IRT). The first test, conducted in February 2009, was to evaluate the current recommended scaling methods for fixed wing on representative rotor airfoils at fixed angle of attack. For this test, scaling was based on the modified Ruff method with scale velocity determined by constant Weber number and water film Weber number. Models were un-swept NACA 0012 wing sections. The reference model had a chord of 91.4 cm and scale model had a chord of 35.6 cm. Reference tests were conducted with velocity of 100 kt (52 m/s), droplet medium volume diameter (MVD) 195 m, and stagnation-point freezing fractions of 0.3 and 0.5 at angle of attack of 5deg and 7deg . It was shown that good ice shape scaling was achieved with constant Weber number for NACA 0012 airfoils with angle of attack up to 7deg . The second test, completed in May 2010, was primarily focused on obtaining transient and steady-state iced aerodynamics, ice accretion and shedding, and thermal icing validation data from an oscillating airfoil section over some selected ranges of icing conditions and blade assembly operational configurations. The model used was a 38.1-cm chord Sikorsky SC2110 airfoil section installed on an airfoil test apparatus with oscillating capability in the IRT. For two test conditions, size and condition scaling were performed. It was shown that good ice shape scaling was achieved for SC2110 airfoil at dynamic pitching motion. The data obtained will be applicable for future main rotor blade and tail rotor blade applications.

  19. Best convective parameterization scheme within RegCM4 to downscale CMIP5 multi-model data for the CORDEX-MENA/Arab domain

    NASA Astrophysics Data System (ADS)

    Almazroui, Mansour; Islam, Md. Nazrul; Al-Khalaf, A. K.; Saeed, Fahad

    2016-05-01

    A suitable convective parameterization scheme within Regional Climate Model version 4.3.4 (RegCM4) developed by the Abdus Salam International Centre for Theoretical Physics, Trieste, Italy, is investigated through 12 sensitivity runs for the period 2000-2010. RegCM4 is driven with European Centre for Medium-Range Weather Forecasts (ECMWF) ERA-Interim 6-hourly boundary condition fields for the CORDEX-MENA/Arab domain. Besides ERA-Interim lateral boundary conditions data, the Climatic Research Unit (CRU) data is also used to assess the performance of RegCM4. Different statistical measures are taken into consideration in assessing model performance for 11 sub-domains throughout the analysis domain, out of which 7 (4) sub-domains give drier (wetter) conditions for the area of interest. There is no common best option for the simulation of both rainfall and temperature (with lowest bias); however, one option each for temperature and rainfall has been found to be superior among the 12 options investigated in this study. These best options for the two variables vary from region to region as well. Overall, RegCM4 simulates large pressure and water vapor values along with lower wind speeds compared to the driving fields, which are the key sources of bias in simulating rainfall and temperature. Based on the climatic characteristics of most of the Arab countries located within the study domain, the drier sub-domains are given priority in the selection of a suitable convective scheme, albeit with a compromise for both rainfall and temperature simulations. The most suitable option Grell over Land and Emanuel over Ocean in wet (GLEO wet) delivers a rainfall wet bias of 2.96 % and a temperature cold bias of 0.26 °C, compared to CRU data. An ensemble derived from all 12 runs provides unsatisfactory results for rainfall (28.92 %) and temperature (-0.54 °C) bias in the drier region because some options highly overestimate rainfall (reaching up to 200 %) and underestimate temperature (reaching up to -1.16 °C). Overall, a suitable option (GLEO wet) is recommended for downscaling the Coupled Model Intercomparison Project Phase 5 (CMIP5) multi-model database using RegCM4 for the CORDEX-MENA/Arab domain for its use in future climate change impact studies.

  20. Lesion size affects diagnostic performance of IOTA logistic regression models, IOTA simple rules and risk of malignancy index in discriminating between benign and malignant adnexal masses.

    PubMed

    Di Legge, A; Testa, A C; Ameye, L; Van Calster, B; Lissoni, A A; Leone, F P G; Savelli, L; Franchi, D; Czekierdowski, A; Trio, D; Van Holsbeke, C; Ferrazzi, E; Scambia, G; Timmerman, D; Valentin, L

    2012-09-01

    To estimate the ability to discriminate between benign and malignant adnexal masses of different size using: subjective assessment, two International Ovarian Tumor Analysis (IOTA) logistic regression models (LR1 and LR2), the IOTA simple rules and the risk of malignancy index (RMI). We used a multicenter IOTA database of 2445 patients with at least one adnexal mass, i.e. the database previously used to prospectively validate the diagnostic performance of LR1 and LR2. The masses were categorized into three subgroups according to their largest diameter: small tumors (diameter < 4 cm; n = 396), medium-sized tumors (diameter, 4-9.9 cm; n = 1457) and large tumors (diameter ≥ 10 cm, n = 592). Subjective assessment, LR1 and LR2, IOTA simple rules and the RMI were applied to each of the three groups. Sensitivity, specificity, positive and negative likelihood ratio (LR+, LR-), diagnostic odds ratio (DOR) and area under the receiver-operating characteristics curve (AUC) were used to describe diagnostic performance. A moving window technique was applied to estimate the effect of tumor size as a continuous variable on the AUC. The reference standard was the histological diagnosis of the surgically removed adnexal mass. The frequency of invasive malignancy was 10% in small tumors, 19% in medium-sized tumors and 40% in large tumors; 11% of the large tumors were borderline tumors vs 3% and 4%, respectively, of the small and medium-sized tumors. The type of benign histology also differed among the three subgroups. For all methods, sensitivity with regard to malignancy was lowest in small tumors (56-84% vs 67-93% in medium-sized tumors and 74-95% in large tumors) while specificity was lowest in large tumors (60-87%vs 83-95% in medium-sized tumors and 83-96% in small tumors ). The DOR and the AUC value were highest in medium-sized tumors and the AUC was largest in tumors with a largest diameter of 7-11 cm. Tumor size affects the performance of subjective assessment, LR1 and LR2, the IOTA simple rules and the RMI in discriminating correctly between benign and malignant adnexal masses. The likely explanation, at least in part, is the difference in histology among tumors of different size. Copyright © 2012 ISUOG. Published by John Wiley & Sons, Ltd.

  1. Rapid, quantitative analysis of ppm/ppb nicotine using surface-enhanced Raman scattering from polymer-encapsulated Ag nanoparticles (gel-colls).

    PubMed

    Bell, Steven E J; Sirimuthu, Narayana M S

    2004-11-01

    Rapid, quantitative SERS analysis of nicotine at ppm/ppb levels has been carried out using stable and inexpensive polymer-encapsulated Ag nanoparticles (gel-colls). The strongest nicotine band (1030 cm(-1)) was measured against d(5)-pyridine internal standard (974 cm(-1)) which was introduced during preparation of the stock gel-colls. Calibration plots of I(nic)/I(pyr) against the concentration of nicotine were non-linear but plotting I(nic)/I(pyr) against [nicotine](x)(x = 0.6-0.75, depending on the exact experimental conditions) gave linear calibrations over the range (0.1-10 ppm) with R(2) typically ca. 0.998. The RMS prediction error was found to be 0.10 ppm when the gel-colls were used for quantitative determination of unknown nicotine samples in 1-5 ppm level. The main advantages of the method are that the gel-colls constitute a highly stable and reproducible SERS medium that allows high throughput (50 sample h(-1)) measurements.

  2. Reduction of turbidity and chromium content of tannery wastewater by electrocoagulation process.

    PubMed

    2018-02-12

    The present study is carried out to remove the chromium and turbidity from tannery wastewater by the electrocoagulationprocess with aluminum electrodes. This experimental study is performed using a batch system. The applied pilot comprises a reactor containing two parallel metal electrodes (Al). The latter are connected as mono polar and a different potential is applied between them. Several working parameters, such as applied potential difference, electrolysis time, active electrode surface, inter-electrode distance and pH of the medium have been studied to achieve higher removal efficiency.The treatment achieved a maximum reduction of 99% for the turbidity and 93% for the chromium under the following conditions: a potential difference: 15V; electrodes surface: 45cm2, inter-electrode distance: 1cm; raw water pH (6.1) and a contact time of 90 min. Considering the obtained efficiency in the present study, electrocoagulation process has the potential to be utilized for the cost-effective removal of pollutants from wastewater.

  3. A simple spectrophotometric determination of meptyldinocap by its hydrolysis.

    PubMed

    Kurup, Sunita; Pillai, Ajai Kumar

    2013-01-01

    A simple spectrophotometric method is proposed for the determination of meptyldinocap (2,4-dinitro-6-octylphenyl crotonate). The method is based on the hydrolysis of meptyldinocap by hydroxylamine solution in alkaline medium to give 2,4-dinitro-6-octylphenol (2,4-DNOP), having maximum absorption at 380 nm. The reaction is found to be instantaneous in presence of ethanol. Beer's law is valid over the concentration range of 1.2-13 microg mL(-1) with molar absorptivity and Sandell's sensitivity of 3.22 x 10(6) L mol(-1) cm(-1) and 0.0001 microg cm(-2) respectively. The limit of detection and quantification were 0.0892 and 0.2703 microg mL(-1), respectively. The tolerance limits of interfering ions are discussed. All variables were studied in order to optimize the reaction conditions. The validity of the method was checked by its simultaneous determination in fruits and water samples and the results were statistically compared with those of a reference method by applying the Student's t-test and F-test.

  4. Interstellar H3O(+) and its relation to the O2 and H2O abundances

    NASA Astrophysics Data System (ADS)

    Phillips, T. G.; van Dishoeck, Ewine F.; Keene, Jocelyn

    1992-11-01

    An interstellar medium study of the three reasonably accessible low-lying submillimeter lines of the H3O(+) molecular ion at 396, 364, and 307 GHz is presented. An analysis of the H3O(+) line ratios shows that under high density (about 10 exp 6 - 10 exp 7/cu cm) and high-temperature (greater than about 50 K), the 396 GHz line is about a factor of two stronger than the 364 GHz line, with the 307 GHz line much weaker. For lower densities, the excitation of the 364 GHz line can be very sensitive to dust radiation pumping, and it is shown that this is the case in Sgr B2, resulting in the 364 GHz line being a factor of 2-3 stronger than the 396 GHz line. Under almost all conditions, the 307 GHz line is weak, the exception being for densities greater than about 10 exp 7/cu cm.

  5. The effect of TNFα secreted from macrophages activated by titanium particles on osteogenic activity regulated by WNT/BMP signaling in osteoprogenitor cells.

    PubMed

    Lee, Sang-Soo; Sharma, Ashish R; Choi, Byung-Soo; Jung, Jun-Sub; Chang, Jun-Dong; Park, Seonghun; Salvati, Eduardo A; Purdue, Edward P; Song, Dong-Keun; Nam, Ju-Suk

    2012-06-01

    Wear particles are the major cause of osteolysis associated with failure of implant following total joint replacement. During this pathologic process, activated macrophages mediate inflammatory responses to increase osteoclastogenesis, leading to enhanced bone resorption. In osteolysis caused by wear particles, osteoprogenitors present along with macrophages at the implant interface may play significant roles in bone regeneration and implant osteointegration. Although the direct effects of wear particles on osteoblasts have been addressed recently, the role of activated macrophages in regulation of osteogenic activity of osteoblasts has scarcely been studied. In the present study, we examined the molecular communication between macrophages and osteoprogenitor cells that may explain the effect of wear particles on impaired bone forming activity in inflammatory bone diseases. It has been demonstrated that conditioned medium of macrophages challenged with titanium particles (Ti CM) suppresses early and late differentiation markers of osteoprogenitors, including alkaline phosphatase (ALP) activity, collagen synthesis, matrix mineralization and expression of osteocalcin and Runx2. Moreover, bone forming signals such as WNT and BMP signaling pathways were inhibited by Ti CM. Interestingly, TNFα was identified as a predominant factor in Ti CM to suppress osteogenic activity as well as WNT and BMP signaling activity. Furthermore, Ti CM or TNFα induces the expression of sclerostin (SOST) which is able to inhibit WNT and BMP signaling pathways. It was determined that over-expression of SOST suppressed ALP activity, whereas the inhibition of SOST by siRNA partially restored the effect of Ti CM on ALP activity. This study highlights the role of activated macrophages in regulation of impaired osteogenic activity seen in inflammatory conditions and provides a potential mechanism for autocrine regulation of WNT and BMP signaling mediated by TNFα via induction of SOST in osteprogenitor cells. Copyright © 2012 Elsevier Ltd. All rights reserved.

  6. A Latest Glacial and Holocene Record From Medicine Lake, Siskiyou County, California: Preliminary Diatom, Pollen, and Sediment Data

    NASA Astrophysics Data System (ADS)

    Starratt, S. W.; Barron, J. A.; Kneeshaw, T.; Phillips, L.; Lowenstern, J.; Wanket, J. A.

    2002-12-01

    Medicine Lake is a small (165 ha), relatively shallow (average 7.3 m), medium- altitude (2,036 m) lake located within the summit caldera of Medicine Lake volcano, a dormant Quaternary shield volcano located in the southern Cascade Range. During September 1999 and 2000, high-resolution bathymetry, seismic-reflection profiles, and sediment cores were collected from the lake. Twenty six samples from core B100NC-1 (water depth 12.6 m; length 226 cm) were analyzed for physical properties, sediment grain size, diatoms, pollen, and total organic carbon (TOC). Using both 14C (AMS) dating and tephrochronology, the sediments at the bottom of the core are estimated to be 11,000 cal yr B.P., thus yielding an estimated average sedimentation rate of about 21 cm/1,000 yr. The lowermost part of the core (226 cm - ~200 cm) records the transition from glacial to interglacial conditions. During the period from about 11,000-7,200 cal yr B.P., lake level fluctuated between deeper oligotrophic conditions with a diatom flora dominated by Cyclotella spp. and shallower intervals with a diverse benthic flora. The relative low abundance (10-15%) of Abies (fir) pollen and relative high abundance (30-40%) of Artemesia (sagebrush) pollen in this interval suggest drier than present-day conditions. The lowest part of this interval (226 cm - 210 cm) is almost devoid of Cyclotella and may represent an ice-covered lake in which only a small benthic flora could exist around the margins of the lake where light penetration was the greatest. The sediments in this interval are relatively low in TOC and are dominated by glacial flour. From about 7,200 cal yr B.P. to the present, conditions have fluctuated between higher lake levels (three intervals) that are dominated by Cyclotella with a reduced number and diversity of benthic taxa, and lower lake levels (two intervals) during which the abundances of Cyclotella decrease to less than 10%. Relative values of Abies and Pinus (pine) pollen are higher during high lake levels, whereas pollen of aquatic taxa (primarily Isoetes [quillwort]) increases in significance at lower lake levels. Total organic carbon is higher during high stands and lower during low stands. Comparison with recently published multi-proxy studies of the Lake Tahoe-Truckee River-Pyramid Lake drainage system suggest that some of the changes in lake level observed at Medicine Lake between about 7,500-4,500 cal yr B.P. may be regional in nature, while fluctuations over the last 4,500 yr probably reflect conditions affecting only the local watershed.

  7. Prognostic Factors Affecting Rotator Cuff Healing After Arthroscopic Repair in Small to Medium-sized Tears.

    PubMed

    Park, Ji Soon; Park, Hyung Jun; Kim, Sae Hoon; Oh, Joo Han

    2015-10-01

    Small and medium-sized rotator cuff tears usually have good clinical and anatomic outcomes. However, healing failure still occurs in some cases. To evaluate prognostic factors for rotator cuff healing in patients with only small to medium-sized rotator cuff tears. Case-control study; Level of evidence, 3. Data were prospectively collected from 339 patients with small to medium-sized rotator cuff tears who underwent arthroscopic repair by a single surgeon between March 2004 and August 2012 and who underwent magnetic resonance imaging or computed tomographic arthrography at least 1 year after surgery. The mean age of the patients was 59.8 years (range, 39-80 years), and the mean follow-up time was 20.8 months (range, 12-66 months). The functional evaluation included the visual analog scale (VAS) for pain, American Shoulder and Elbow Surgeons score, Constant-Murley score, and Simple Shoulder Test. Postoperative VAS for pain and functional scores improved significantly compared with preoperative values (P < .001). Forty-five healing failures occurred (13.3%), and fatty degeneration of the infraspinatus muscle, tear size (anteroposterior dimension), and age were significant factors affecting rotator cuff healing (P < .001, = .018, and = .011, respectively) in multivariate logistic regression analysis. Grade II and higher infraspinatus fatty degeneration correlated with a higher failure rate. The failure rate was also significantly higher in patients with a tear >2 cm in size (34.2%) compared with patients with a tear ≤2 cm (10.6%) (P < .001). A receiver operating characteristic curve was used to determine the predictive cut-off value for the oldest age and the largest tear size for successful healing, which were calculated as 69 years and 2 cm, respectively, with a specificity of 90%. In small to medium-sized rotator cuff tears, grade II fatty degeneration of the infraspinatus muscle according to the Goutallier classification could be a reference point for successful healing, and anatomic outcomes might be better if repair is performed before the patient is 69 years old and the tear size exceeds 2 cm. © 2015 The Author(s).

  8. Laser biostimulation of articular cartilage: in vitro evaluation

    NASA Astrophysics Data System (ADS)

    Jia, Yali; Guo, Zhouyi; Yang, Xiaohong; Zeng, Chang-Chun

    2004-07-01

    In the orthopaedic field, the repair of ariticular cartilage is still a difficult problem, because of the physiological characters of cartilaginous tissues and chondrocytes. To find an effective method of stimulating their regeneration, this in vitro study focuses on the biostimulation of rabbit articular chondrocytes by low-power He-Ne laser. The articular chondrocytes isolated from the cartilage of the medial condyle of the femur of the rabbit were incubated in HamF12 medium. The second passage culture were spread on 24 petri dishes and were irradiated with laser at power density of 2 - 12 mW/cm2 for 6.5 minutes, corresponding to the energy density of 1-6 J/cm2. Laser treatment was performed three times at a 24-hour interval. After lasering, incubation was continued for 24 hours. Non-irradiated cells were kept under the same conditions as the irradiated ones. The cell proliferation activity was evaluated with a XTT colorimetric method. Irradiation of 4 - 6 J/cm2 revealed a considerably higher cell proliferation activity comparing to control cultures. Thereinto, the energy density of 4 and 5 J/cm2 remarkably increased cell growth (P<0.01). The present study showed that a particular laser irradiation stimulates articular chondrocytes proliferation. These findings might be clinically relevant, indicating that low-power laser irradiation treatment is likely to achieve the repair of articular cartilage in clinic.

  9. Hyporheic zone as a bioreactor: sediment heterogeneity influencing biogeochemical processes

    NASA Astrophysics Data System (ADS)

    Perujo, Nuria; Romani, Anna M.; Sanchez-Vila, Xavier

    2017-04-01

    Mediterranean fluvial systems are characterized by frequent periods of low flow or even drought. During low flow periods, water from wastewater treatment plants (WWTPs) is proportionally large in fluvial systems. River water might be vertically transported through the hyporheic zone, and then porous medium acts as a complementary treatment system since, as water infiltrates, a suite of biogeochemical processes occurs. Subsurface sediment heterogeneity plays an important role since it influences the interstitial fluxes of the medium and drives biomass growing, determining biogeochemical reactions. In this study, WWTP water was continuously infiltrated for 3 months through two porous medium tanks: one consisting of 40 cm of fine sediment (homogeneous); and another comprised of two layers of different grain size sediments (heterogeneous), 20 cm of coarse sediment in the upper part and 20 cm of fine one in the bottom. Several hydrological, physicochemical and biological parameters were measured periodically (weekly at the start of the experiment and biweekly at the end). Analysed parameters include dissolved nitrogen, phosphorus, organic carbon, and oxygen all measured at the surface, and at 5, 20 and 40 cm depth. Variations in hydraulic conductivity with time were evaluated. Sediment samples were also analysed at three depths (surface, 20 and 40 cm) to determine bacterial density, chlorophyll content, extracellular polymeric substances, and biofilm function (extracellular enzyme activities and carbon substrate utilization profiles). Preliminary results suggest hydraulic conductivity to be the main driver of the differences in the biogeochemical processes occurring in the subsurface. At the heterogeneous tank, a low nutrient reduction throughout the whole medium is measured. In this medium, high hydraulic conductivity allows for a large amount of infiltrating water, but with a small residence time. Since some biological processes are largely time-dependent, small water residence time results in low nutrient reduction. Moreover, high nitrification and low ammonium concentration in the interface of the two grain-size layers are measured, probably related to high dissolved oxygen concentration at the coarse-fine sediment interface, further promoting accumulation of bacteria and algae. In contrast, the homogeneous tank shows low dissolved oxygen values and high denitrification in depth which could be related to lower overall hydraulic conductivity, as compared to the heterogeneous tank. The preliminary analysis of our results indicates a key role of hydraulic conductivity on biogeochemical processes in the porous medium but, at the same time that it is strongly interacting with sediment grain-size distribution and the development of biofilm. The final scope of this study is to know the interactions between physicochemical and biological components in sediments in order to understand in detail the biogeochemical processes occurring.

  10. In Vitro Propagation of Sambong (Blumea balsamifera Linn.).

    PubMed

    Soriano, Thelma L; Cangao, Evangelina C

    2016-01-01

    Terminal shoot tips of sambong (Blumea balsamifera Linn.) are cultured to initiate and regenerate shoots on Murashige and Skoog (MS) medium containing 1.0 mg/L benzyl adenine (BA). After 1 month, shoots, usually 4.5 cm long are separated and subcultured for multiplication. Regenerated shoots, about 6 cm long are rooted on MS medium supplemented with 1.0 mg/L naphthalene acetic acid (NAA). Exposure of shoots to high humidity for the first 2 weeks and equal proportion (1:1:1) of sterile sand, compost, and coir dust as potting mix favors the development of whole sambong plants. Young shoots from in vitro-derived sambong plants could also be used for propagation.

  11. Interleukin-10 Contributes to Therapeutic Effect of Mesenchymal Stem Cells for Acute Liver Failure via Signal Transducer and Activator of Transcription 3 Signaling Pathway

    PubMed Central

    Ma, Hu-Cheng; Wang, Xin; Wu, Min-Na; Zhao, Xin; Yuan, Xian-Wen; Shi, Xiao-Lei

    2016-01-01

    Background: Mesenchymal stem cells (MSCs) transplantation has been proven to have therapeutic potential for acute liver failure (ALF). However, the mechanism remains controversial. Recently, modulation of inflammation by MSCs has been regarded as a crucial mechanism. The aim of the present study was to explore the soluble cytokines secreted by MSCs and their therapeutic effects in ALF. Methods: MSCs isolated from Sprague-Dawley rats were identified by fluorescence-activated cell sorting analysis. Conditioned medium derived from MSCs (MSCs-CM) was collected and analyzed by a cytokine microarray. MSCs and MSCs-CM were transplanted into rats with D-galactosamine-induced ALF. Liver function, survival rate, histology, and inflammatory factors were determined. Exogenous recombinant rat interleukin (IL)-10, anti-rat IL-10 antibody, and AG490 (signal transducer and activator of transcription 3 [STAT3] signaling pathway inhibitor) were administered to explore the therapeutic mechanism of MSCs-CM. Statistical analysis was performed with SPSS version 19.0, and all data were analyzed by the independent-sample t-test. Results: There are statistical differences of the survival curve between ALF+MSCs group and ALF+Dulbecco's modified Eagle's medium (DMEM) group, as well as ALF+MSCs-CM group and ALF+DMEM group (all P < 0.05). Serum alanine aminotransferase (ALT) level in the ALF+MSCs and ALF+MSCs-CM groups was lower than that in the ALF+DMEM group (865.53±52.80 vs. 1709.75±372.12 U/L and 964.72±414.59 vs. 1709.75±372.12 U/L, respectively, all P < 0.05); meanwhile, serum aspartate aminotransferase (AST) level in the ALF+MSCs and ALF+MSCs-CM groups was lower than that in the ALF+DMEM group (2440.83±511.94 vs. 4234.35±807.30 U/L and 2739.83±587.33 vs. 4234.35±807.30 U/L, respectively, all P < 0.05). Furthermore, MSCs or MSCs-CM treatment significantly reduced serum interferon-γ (IFN-γ), IL-1β, IL-6 levels and increased serum IL-10 level compared with DMEM (all P < 0.05). Proteome profile analysis of MSCs-CM indicated the presence of anti-inflammatory factors and IL-10 was the most distinct. Blocking of IL-10 confirmed the therapeutic significance of this cytokine. Phosphorylated STAT3 was upregulated after IL-10 infusion and inhibition of STAT3 by AG490 reversed the therapeutic effect of IL-10. Conclusions: The factors released by MSCs, especially IL-10, have the potential for therapeutic recovery of ALF, and the STAT3 signaling pathway may mediate the anti-inflammatory effect of IL-10. PMID:27064043

  12. Better Rooting Procedure to Enhance Survival Rate of Field Grown Malaysian Eksotika Papaya Transformed with 1-Aminocyclopropane-1-Carboxylic Acid Oxidase Gene

    PubMed Central

    Sekeli, Rogayah; Abdullah, Janna Ong; Namasivayam, Parameswari; Muda, Pauziah; Abu Bakar, Umi Kalsom

    2013-01-01

    A high survival rate for transformed papaya plants when transferred to the field is useful in the quest for improving the commercial quality traits. We report in this paper an improved rooting method for the production of transformed Malaysian Eksotika papaya with high survival rate when transferred to the field. Shoots were regenerated from embryogenic calli transformed with antisense and RNAi constructs of 1-aminocyclopropane-1-carboxylic acid oxidase (ACO) genes using the Agrobacterium tumefaciens-mediated transformation method. Regenerated transformed shoots, each measuring approximately 3-4 cm in height, were cultured in liquid half-strength Murashige and Skoog (MS) medium or sterile distilled water, and with either perlite or vermiculite supplementation. All the culturing processes were conducted either under sterile or nonsterile condition. The results showed that rooting under sterile condition was better. Shoots cultured in half-strength MS medium supplemented with vermiculite exhibited a 92.5% rooting efficiency while perlite showed 77.5%. The survival rate of the vermiculite-grown transformed papaya plantlets after transfer into soil, contained in polybags, was 94%, and the rate after transfer into the ground was 92%. Morpho-histological analyses revealed that the tap roots were more compact, which might have contributed to the high survival rates of the plantlets. PMID:25969786

  13. Better rooting procedure to enhance survival rate of field grown malaysian eksotika papaya transformed with 1-aminocyclopropane-1-carboxylic Acid oxidase gene.

    PubMed

    Sekeli, Rogayah; Abdullah, Janna Ong; Namasivayam, Parameswari; Muda, Pauziah; Abu Bakar, Umi Kalsom

    2013-01-01

    A high survival rate for transformed papaya plants when transferred to the field is useful in the quest for improving the commercial quality traits. We report in this paper an improved rooting method for the production of transformed Malaysian Eksotika papaya with high survival rate when transferred to the field. Shoots were regenerated from embryogenic calli transformed with antisense and RNAi constructs of 1-aminocyclopropane-1-carboxylic acid oxidase (ACO) genes using the Agrobacterium tumefaciens-mediated transformation method. Regenerated transformed shoots, each measuring approximately 3-4 cm in height, were cultured in liquid half-strength Murashige and Skoog (MS) medium or sterile distilled water, and with either perlite or vermiculite supplementation. All the culturing processes were conducted either under sterile or nonsterile condition. The results showed that rooting under sterile condition was better. Shoots cultured in half-strength MS medium supplemented with vermiculite exhibited a 92.5% rooting efficiency while perlite showed 77.5%. The survival rate of the vermiculite-grown transformed papaya plantlets after transfer into soil, contained in polybags, was 94%, and the rate after transfer into the ground was 92%. Morpho-histological analyses revealed that the tap roots were more compact, which might have contributed to the high survival rates of the plantlets.

  14. Intestinal bicarbonate secretion in Amphiuma measured by pH stat in vitro: relationship with metabolism and transport of sodium and chloride ions.

    PubMed Central

    Imon, M A; White, J F

    1981-01-01

    1. Isolated Amphiuma small intestine exposed on both surfaces to buffered or unbuffered media generated gradients of pH under short-circuited conditions consistent with secretion of HCO3(-). 2. When unbuffered mucosal medium was maintained at pH 7.4 by addition of acid, alkalinization of the mucosal medium occurred at a rate of 1-2 microequiv/hr cm2 under short-circuit conditions (Isc) and was reduced by anoxia, acetazolamide or removal of CO2. 3. The rate of HCO3(-) secretion (JHCO3(-)) was reduced at a mucosal pH above or below 7.4 and was proportional to serosal HCO3(-). 4. JHCO3(-) was reduced in Na+-free (choline) and Cl-free (SO4(2-) media and after exposure to the stilbene SITS. 5. The difference JHCO3(-)--Isc was consistent with net Cl- absorption. 6. The tissue resistance (Rt) was elevated upon exposure to serosal HCO3(-) and lowered by mucosal HCO3(-). 7. The intestinal mucosa exhibited carbonic anhydrase activity that was sensitive to ethoxazolamide. 8. It is concluded that HCO3(-) secretion is active, influenced by intracellular carbonic anhydrase activity and coupled to Cl- and possibly Na+ absorption. PMID:7310697

  15. Intestinal bicarbonate secretion in Amphiuma measured by pH stat in vitro: relationship with metabolism and transport of sodium and chloride ions.

    PubMed

    Imon, M A; White, J F

    1981-05-01

    1. Isolated Amphiuma small intestine exposed on both surfaces to buffered or unbuffered media generated gradients of pH under short-circuited conditions consistent with secretion of HCO3(-). 2. When unbuffered mucosal medium was maintained at pH 7.4 by addition of acid, alkalinization of the mucosal medium occurred at a rate of 1-2 microequiv/hr cm2 under short-circuit conditions (Isc) and was reduced by anoxia, acetazolamide or removal of CO2. 3. The rate of HCO3(-) secretion (JHCO3(-)) was reduced at a mucosal pH above or below 7.4 and was proportional to serosal HCO3(-). 4. JHCO3(-) was reduced in Na+-free (choline) and Cl-free (SO4(2-) media and after exposure to the stilbene SITS. 5. The difference JHCO3(-)--Isc was consistent with net Cl- absorption. 6. The tissue resistance (Rt) was elevated upon exposure to serosal HCO3(-) and lowered by mucosal HCO3(-). 7. The intestinal mucosa exhibited carbonic anhydrase activity that was sensitive to ethoxazolamide. 8. It is concluded that HCO3(-) secretion is active, influenced by intracellular carbonic anhydrase activity and coupled to Cl- and possibly Na+ absorption.

  16. TRANSVERSE ELECTRIC IMPEDANCE OF NITELLA

    PubMed Central

    Curtis, Howard J.; Cole, Kenneth S.

    1937-01-01

    Alternating current measurements have been taken on single Nitella cells over a frequency range from 30 to 2,500,000 cycles per second with the current flow perpendicular to the axis of the cell. The measuring cells were so constructed that electrolytes of any desired concentration could be circulated during the course of the measurements. The cellulose wall which surrounds the cell is found to play an important part in the interpretation of the results obtained. In a mature cell, this cellulose has a specific resistance of about 1000 ohm cm. which is independent of the medium in which the cell is suspended. The thickness of the wall is computed to be about 10 µ. The cell membrane is found to be virtually non-conducting, and to have a capacity of 0.94 µf./cm.2 ± 10 per cent and a phase angle of 80° ± 4°. The specific resistances of the sap were difficult to compute from data on living cells and were unsatisfactory because they were very much dependent upon the medium, while measurements on extracted sap gave 58 ohm cm. ± 8 per cent which was independent of the medium. There are indications that the chloroplasts have impedance properties similar to those of living cells. PMID:19873046

  17. Differential effects of the steaming time and frequency for manufactured red Liriope platyphylla on nerve growth factor secretion ability, nerve growth factor receptor signaling pathway and regulation of calcium concentration.

    PubMed

    Choi, Sun Il; Goo, Jun Seo; Kim, Ji Eun; Nam, So Hee; Hwang, In Sik; Lee, Hye Ryun; Lee, Young Ju; Son, Hong Joo; Lee, Hee Seob; Lee, Jong Sup; Kim, Hak Jin; Hwang, Dae Youn

    2012-11-01

    The herb Liriope platyphylla (LP) has been considered to have curative properties for diabetes, asthma and neurodegenerative disorders. To examine the effects of steaming time and frequency of manufactured red LP (RLP) on the nerve growth factor (NGF) secretion ability and NGF receptor signaling pathway, the NGF concentration, cell differentiation, NGF signaling pathway and calcium concentration were analyzed in neuronal cells treated with several types of LPs manufactured under different conditions. The maximum NGF secretion was observed in B35 cells treated with 50 µg/ml LP extract steamed for 9 h (9-SLP) and with two repeated steps (3 h steaming and 24 h air-dried) carried out 7 times (7-SALP). No significant changes in viability were detected in any of the cells treated with the various LPs, with the exception of 0-SLP and 0-SALP. In addition, PC12 cell differentiation was induced by treatment with the NGF-containing conditional medium (CM) collected from the RLP-treated cells. The levels of TrkA and extracellular signal-regulated kinase (ERK) phosphorylation in the high affinity NGF receptor signaling pathway were significantly higher in the cells treated with 3-SLP or 1-SALP/3-SALP CM compared with those treated with the vehicle CM. In the low affinity NGF receptor pathway, the expression levels of most components were higher in the 9-, 15- and 24-SALP CM-treated cells compared with the vehicle CM-treated cells. However, this level was significantly altered in cells treated with 3-SALP CM. Furthermore, an examination of the RLP function on calcium regulation revealed that only the LP- or RLP-treated cells exhibited changes in intracellular and extracellular calcium levels. RLP induced a significant decrease in the intracellular calcium levels and an increase in the extracellular calcium levels. These results suggest the possibility that steaming-processed LP may aid in the relief of neurodegenerative diseases through the NGF secretion ability and NGF signaling pathway.

  18. Extremely Low Frequency Electromagnetic Field from Convective Air Warming System on Temperature Selection and Distance.

    PubMed

    Cho, Kwang Rae; Kim, Myoung-Hun; Ko, Myoung Jin; Jung, Jae Wook; Lee, Ki Hwa; Park, Yei-Heum; Kim, Yong Han; Kim, Ki Hoon; Kim, Jin Soo

    2014-12-01

    Hypothermia generates potentially severe complications in operating or recovery room. Forced air warmer is effective to maintain body temperature. Extremely low frequency electromagnetic field (ELF-EMF) is harmful to human body and mainly produced by electronic equipment including convective air warming system. We investigated ELF-EMF from convective air warming device on various temperature selection and distance for guideline to protect medical personnel and patients. The intensity of ELF-EMF was measured as two-second interval for five minutes on various distance (0.1, 0.2, 0.3, 0.5 and 1meter) and temperature selection (high, medium, low and ambient). All of electrical devices were off including lamp, computer and air conditioner. Groups were compared using one-way ANOVA. P<0.05 was considered significant. Mean values of ELF-EMF on the distance of 30 cm were 18.63, 18.44, 18.23 and 17.92 milligauss (mG) respectively (high, medium, low and ambient temperature set). ELF-EMF of high temperature set was higher than data of medium, low and ambient set in all the distances. ELF-EMF from convective air warming system is higher in condition of more close location and higher temperature. ELF-EMF within thirty centimeters exceeds 2mG recommended by Swedish TCO guideline.

  19. Effect of air velocity and direction for indirect evaporative cooling in tropical area

    NASA Astrophysics Data System (ADS)

    Ayodha Ajiwiguna, Tri; Nugraha Rismi, Fadhlin; Ramdlan Kirom, Mukhammad

    2017-06-01

    In this research, experimental study of heat absorption rate caused by indirect evaporative cooling is performed by varying the velocity and direction of air. The ambient is at average temperature and relative humidity of 28.7 °C and 78% respectively. The experiment is conducted by attaching wet medium on the top of material reference plate with the dimension of 14 x 8 cm with 5 mm thickness. To get evaporative cooling effect, the air flow is directed to the wet medium with velocity from 1.6 m/s to 3.4 m/s with the increment of 0.2 m/s. The direction of air is set 0° (parallel), 45° (inclined), and 90° (perpendicular) to the wet medium surface. While the experiment is being performed, the air temperature, top and bottom of plate temperature are measured simultaneously after steady state condition is established. Based on the measurement result, heat absorption is calculated by analysing the heat conduction on the material reference. The result shows that the heat absorption rate is increased by higher velocity. Perpendicular direction of air flow results the highest cooling capacity compared with other direction. The maximum heat absorption rate is achieved at 13.9 Watt with 3.4 m/s velocity and perpendicular direction of air.

  20. SPH-4 U.S. Army Flight Helmet Performance 1972-1983

    DTIC Science & Technology

    1984-11-01

    epoxy resin and fiberglass cloth. 2. Liner -Energy-absorbing 1.3 cm thick expanded polystyrene with a density of 0.08 gm/cm 3 3. Suspension - With...thick epoxy or polyester resin and fiberglass cloth layup provided in small, medium, and large siaeso b. Liner- Energy-absorbing 1.3 cm thick expanded ... polystyrene -foam with density of .08 gm/cu3. c ’ pension- Provided by three leather-covered foam pads located at the front, crown, and rear of the

  1. Wharton's jelly mesenchymal stromal cells have contrasting effects on proliferation and phenotype of cancer stem cells from different subtypes of lung cancer

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

    Vulcano, Francesca, E-mail: francesca.vulcano@iss.it; Milazzo, Luisa, E-mail: luisa.milazzo@iss.it; Ciccarelli, Carmela, E-mail: carmela.ciccarelli@univaq.it

    Studies on the role of multipotent mesenchymal stromal cells (MSC) on tumor growth have reported both a tumor promoting and a suppressive effect. The aim of the present study was to determine the effect of MSC isolated from Wharton's jelly of umbilical cord (WJMSC) on lung cancer stem cells (LCSC) derived from human lung tumors: two adenocarcinomas (AC) and two squamous cell carcinomas (SCC). LCSC derived from SCC and AC expressed, to varying extents, the more relevant stem cell markers. The effect of WJMSC on LCSC was investigated in vitro using conditioned medium (WJ-CM): a proliferation increase in AC-LCSC wasmore » observed, with an increase in the ALDH+ and in the CD133+ cell population. By contrast, WJ-CM hampered the growth of SCC-LCSC, with an increase in the pre-G1 phase indicating the induction of apoptosis. Furthermore, the ALDH+ and CD133+ population was also reduced. In vivo, subcutaneous co-transplantation of AC-LCSC/WJMSC generated larger tumors than AC-LCSC alone, characterized by an increased percentage of CD133+ and CD166+ cells. By contrast, co-transplantation of WJMSC and SCC-LCSC did not affect the tumor size. Our results strongly suggest that WJMSC exert, both in vitro and in vivo, contrasting effects on LCSC derived from different lung tumor subtypes. - Highlights: • CM from WJMSC induces apoptosis of SCC-LCSC and reduction of ALDH+ and CD133+ cells. • Specificity of SCC-LCSC inhibition by WJ-CM is proved by the use of a CM from NHDF. • WJ-CM enhance AC-LCSC proliferation and increase CD133+ and ALDH+ cell fractions. • Coinjection of WJMSC with AC-LCSC increase tumor growth with SCC-LCSC has no effect.« less

  2. The secretome of endothelial progenitor cells promotes brain endothelial cell activity through PI3-kinase and MAP-kinase.

    PubMed

    Di Santo, Stefano; Seiler, Stefanie; Fuchs, Anna-Lena; Staudigl, Jennifer; Widmer, Hans Rudolf

    2014-01-01

    Angiogenesis and vascular remodelling are crucial events in tissue repair mechanisms promoted by cell transplantation. Current evidence underscores the importance of the soluble factors secreted by stem cells in tissue regeneration. In the present study we investigated the effects of paracrine factors derived from cultured endothelial progenitor cells (EPC) on rat brain endothelial cell properties and addressed the signaling pathways involved. Endothelial cells derived from rat brain (rBCEC4) were incubated with EPC-derived conditioned medium (EPC-CM). The angiogenic response of rBCEC4 to EPC-CM was assessed as effect on cell number, migration and tubular network formation. In addition, we have compared the outcome of the in vitro experiments with the effects on capillary sprouting from rat aortic rings. The specific PI3K/AKT inhibitor LY294002 and the MEK/ERK inhibitor PD98059 were used to study the involvement of these two signaling pathways in the transduction of the angiogenic effects of EPC-CM. Viable cell number, migration and tubule network formation were significantly augmented upon incubation with EPC-CM. Similar findings were observed for aortic ring outgrowth with significantly longer sprouts. The EPC-CM-induced activities were significantly reduced by the blockage of the PI3K/AKT and MEK/ERK signaling pathways. Similarly to the outcome of the rBCEC4 experiments, inhibition of the PI3K/AKT and MEK/ERK pathways significantly interfered with capillary sprouting induced by EPC-CM. The present study demonstrates that EPC-derived paracrine factors substantially promote the angiogenic response of brain microvascular endothelial cells. In addition, our findings identified the PI3K/AKT and MEK/ERK pathways to play a central role in mediating these effects.

  3. Guiding of High Laser Intensities in Long Plasma Channels

    NASA Astrophysics Data System (ADS)

    Levin, M.; Eisenmann, S.; Palchan, T.; Zigler, A.; Sugiyama, K.; Nakajima, K.; Kaganovich, D.; Hubbard, R. F.; Ting, A.; Gordon, D. F.; Sprangle, P.; Fraenkel, M.; Maman, S.; Henis, Z.

    Plasma channels have been widely used to guide intense laser pulses over many Rayleigh lengths. Using optimized segmented capillary discharges, we demonstrated guided propagation of ultra short (100 fs) high intensity (1016 W/cm-2, limited by the laser system) pulses over distances up to 12.6 cm and intensities above 1018W/cm2 for 1.5cm boron nitride capillary. Both radial and longitudinal density profiles of plasma channels were studied under various discharge conditions. A new diagnostic technique is presented in which the transport of a guided laser pulse at different delay times from the initiation of the discharge is sampled on a single discharge shot. Using external, 10 nsec Nd YAG laser of several tenths of milijoules to ignite polyethylene capillaries we have demonstrated channels of various length in density range of 1017 - 1019 cm-3 and up to 25% deep. The longitudinal profiles were found to be remarkably uniform in both short and long capillaries. The Boron Nitride capillary has provided a guiding medium that can withstand more than 1000 shots. Using these capillaries we have guided laser intensities above 1018W/cm2. The laser ignition of capillary discharge provided reliable almost jitter free approach. The concerns related to influence of relatively high current density flow through capillary on the injected electrons were studied extensively by us both theoretically and experimentally using a simple injection method. The method is based on the interaction of a high intensity laser pulse with a thin wire placed near capillary entrance. The influence of magnetic fields was found to be insignificant. Using this method we have studied transport of electrons though capillary discharge.

  4. In vitro characterization of human hair follicle dermal sheath mesenchymal stromal cells and their potential in enhancing diabetic wound healing.

    PubMed

    Ma, Dongrui; Kua, Jonah Ee Hsiang; Lim, Wee Keng; Lee, Seng Teik; Chua, Alvin Wen Choong

    2015-08-01

    Little is published on the characterization and therapeutic potential of human mesenchymal cells derived from hair follicle (HF) dermal sheath (DS). In this study, we isolated and characterized HF DS-mesenchymal stromal cells (DS-MSCs) with respect to the bone marrow mesenchymal stromal cells (BM-MSCs). We further tested if DS-MSC-conditioned medium (CM), like what was previously reported for BM-MSC CM, has superior wound-healing properties, in both in vitro and in vivo wound models compared with skin fibroblast CM. DS-MSCs were isolated from HF and cultured in vitro to assess long-term growth potential, colony-forming efficiency (CFE), expression of CD surface markers and differentiation potential. The cytokine expression of DS-MSC CM was determined through an antibody-based protein array analysis. The wound-healing effects of the CM were tested in vitro with the use of human cell cultures and in vivo with the use of a diabetic mouse wound model. In vitro results revealed that DS-MSCs have high growth capacity and CFE while displaying some phenotypes similar to BM-MSCs. DS-MSCs strongly expressed many surface markers expressed in BM-MSCs and could also differentiate into osteoblasts, chondrocytes and adipocytes. DS-MSCs secreted significantly higher proportions of paracrine factors such as interleukin-6 (IL-6), IL-8 and growth-related oncogene. DS-MSC-CM demonstrated enhanced wound-healing effects on human skin keratinocytes, fibroblasts and endothelial cells in vitro, and the wound-healing time in diabetic mice was found to be shorter, compared with vehicle controls. Human HF DS stromal cells demonstrated MSC-like properties and might be an alternative source for therapeutic use in wound healing. Copyright © 2015 International Society for Cellular Therapy. Published by Elsevier Inc. All rights reserved.

  5. Very high cell density perfusion of CHO cells anchored in a non-woven matrix-based bioreactor.

    PubMed

    Zhang, Ye; Stobbe, Per; Silvander, Christian Orrego; Chotteau, Véronique

    2015-11-10

    Recombinant Chinese Hamster Ovary (CHO) cells producing IgG monoclonal antibody were cultivated in a novel perfusion culture system CellTank, integrating the bioreactor and the cell retention function. In this system, the cells were harbored in a non-woven polyester matrix perfused by the culture medium and immersed in a reservoir. Although adapted to suspension, the CHO cells stayed entrapped in the matrix. The cell-free medium was efficiently circulated from the reservoir into- and through the matrix by a centrifugal pump placed at the bottom of the bioreactor resulting in highly homogenous concentrations of the nutrients and metabolites in the whole system as confirmed by measurements from different sampling locations. A real-time biomass sensor using the dielectric properties of living cells was used to measure the cell density. The performances of the CellTank were studied in three perfusion runs. A very high cell density measured as 200 pF/cm (where 1 pF/cm is equivalent to 1 × 10(6)viable cells/mL) was achieved at a perfusion rate of 10 reactor volumes per day (RV/day) in the first run. In the second run, the effect of cell growth arrest by hypothermia at temperatures lowered gradually from 37 °C to 29 °C was studied during 13 days at cell densities above 100 pF/cm. Finally a production run was performed at high cell densities, where a temperature shift to 31 °C was applied at cell density 100 pF/cm during a production period of 14 days in minimized feeding conditions. The IgG concentrations were comparable in the matrix and in the harvest line in all the runs, indicating no retention of the product of interest. The cell specific productivity was comparable or higher than in Erlenmeyer flask batch culture. During the production run, the final harvested IgG production was 35 times higher in the CellTank compared to a repeated batch culture in the same vessel volume during the same time period. Copyright © 2015 The Authors. Published by Elsevier B.V. All rights reserved.

  6. Successful Sampling Strategy Advances Laboratory Studies of NMR Logging in Unconsolidated Aquifers

    NASA Astrophysics Data System (ADS)

    Behroozmand, Ahmad A.; Knight, Rosemary; Müller-Petke, Mike; Auken, Esben; Barfod, Adrian A. S.; Ferré, Ty P. A.; Vilhelmsen, Troels N.; Johnson, Carole D.; Christiansen, Anders V.

    2017-11-01

    The nuclear magnetic resonance (NMR) technique has become popular in groundwater studies because it responds directly to the presence and mobility of water in a porous medium. There is a need to conduct laboratory experiments to aid in the development of NMR hydraulic conductivity models, as is typically done in the petroleum industry. However, the challenge has been obtaining high-quality laboratory samples from unconsolidated aquifers. At a study site in Denmark, we employed sonic drilling, which minimizes the disturbance of the surrounding material, and extracted twelve 7.6 cm diameter samples for laboratory measurements. We present a detailed comparison of the acquired laboratory and logging NMR data. The agreement observed between the laboratory and logging data suggests that the methodologies proposed in this study provide good conditions for studying NMR measurements of unconsolidated near-surface aquifers. Finally, we show how laboratory sample size and condition impact the NMR measurements.

  7. Studies on the resistance/reactivation of Giardia muris cysts and Cryptosporidium parvum oocysts exposed to medium-pressure ultraviolet radiation.

    PubMed

    Belosevic, M; Craik, S A; Stafford, J L; Neumann, N F; Kruithof, J; Smith, D W

    2001-10-16

    The ex vivo and in vivo reactivation of Giardia muris cysts and Cryptosporidium parvum oocysts after exposure to different doses of ultraviolet (UV) radiation was determined using animal infectivity. The infectivity of UV-treated parasites stored for 1-4 days (G. muris) or 1-17 days (C. parvum) at room temperature in the dark was similar to that of organisms administered immediately after UV treatment, indicating that the parasites did not reactivate ex vivo. In contrast, we observed in vivo reactivation of G. muris in three of seven independent animal infectivity experiments, when parasites were treated with relatively low doses of medium-pressure UV (<25 mJ/cm(2)). Our observations indicate that G. muris cysts and C. parvum oocysts exposed to medium-pressure UV doses of 60 mJ/cm(2) or higher did not exhibit resistance to and/or reactivation following treatment. This suggests that when appropriate doses of UV are used, significant and permanent inactivation of these parasites may be achieved.

  8. Defined Essential 8™ Medium and Vitronectin Efficiently Support Scalable Xeno-Free Expansion of Human Induced Pluripotent Stem Cells in Stirred Microcarrier Culture Systems

    PubMed Central

    Badenes, Sara M.; Fernandes, Tiago G.; Cordeiro, Cláudia S. M.; Boucher, Shayne; Kuninger, David; Vemuri, Mohan C.; Diogo, Maria Margarida; Cabral, Joaquim M. S.

    2016-01-01

    Human induced pluripotent stem (hiPS) cell culture using Essential 8™ xeno-free medium and the defined xeno-free matrix vitronectin was successfully implemented under adherent conditions. This matrix was able to support hiPS cell expansion either in coated plates or on polystyrene-coated microcarriers, while maintaining hiPS cell functionality and pluripotency. Importantly, scale-up of the microcarrier-based system was accomplished using a 50 mL spinner flask, under dynamic conditions. A three-level factorial design experiment was performed to identify optimal conditions in terms of a) initial cell density b) agitation speed, and c) to maximize cell yield in spinner flask cultures. A maximum cell yield of 3.5 is achieved by inoculating 55,000 cells/cm2 of microcarrier surface area and using 44 rpm, which generates a cell density of 1.4x106 cells/mL after 10 days of culture. After dynamic culture, hiPS cells maintained their typical morphology upon re-plating, exhibited pluripotency-associated marker expression as well as tri-lineage differentiation capability, which was verified by inducing their spontaneous differentiation through embryoid body formation, and subsequent downstream differentiation to specific lineages such as neural and cardiac fates was successfully accomplished. In conclusion, a scalable, robust and cost-effective xeno-free culture system was successfully developed and implemented for the scale-up production of hiPS cells. PMID:26999816

  9. Establishment of a medium-scale mosquito facility: tests on mass production cages for Aedes albopictus (Diptera: Culicidae).

    PubMed

    Zhang, Dongjing; Li, Yongjun; Sun, Qiang; Zheng, Xiaoying; Gilles, Jeremie R L; Yamada, Hanano; Wu, Zhongdao; Xi, Zhiyong; Wu, Yu

    2018-03-19

    Mass egg production is an important component of Aedes albopictus mosquito control programs, such as the sterile insect technique and incompatible insect technique, which requires the releases of large number of sterile males. Developing standard operating procedures and optimized cages for adult maintenance of Ae. albopictus can improve the mass rearing efficiency. Three different sex ratios of females to males with a total number of 4,000 mosquitoes were tested by evaluating the insemination rate, egg production (total number of eggs per cage), female fecundity and egg hatch rate in small cage (30 × 30 × 30 cm). Blood meals with adenosine triphosphate (ATP, 0.05 g/ml), cage structures (Big cage A: 90 × 30 × 30 cm; Big cage B: 90 × 30 × 50 cm or 90 × 50 × 30 cm) and rearing densities (12,000, 16,000 and 20,000 mosquitoes, corresponding to 0.9 cm 2 /mosquito, 0.675 cm 2 /mosquito and 0.54 cm 2 /mosquito, respectively) were also tested and evaluated on the basis of egg production, female fecundity and egg hatch rate. An adult rearing unit holding 15 of Big cage A with optimal egg production was designed to produce 10 million eggs per rearing cycle in a 1.8 m 2 space. Female to male ratios at 3:1 in small cages resulted in higher egg production but did not affect insemination rate, female fecundity and egg hatch rate. A concentration of 0.05 g/ml of ATP added to blood meals improved the blood-feeding frequency and thus increased the overall egg production per cage. Cage structures affected the egg production per cage, but not egg hatch rate. A medium rearing density at 0.675 cm 2 /mosquito (16,000 mosquitoes) resulted in higher egg production compared to both low and high densities. An adult rearing unit for Ae. albopictus on the basis of Big cage A has been developed with the capacity of producing 10 million eggs within 15 days. Our results have indicated that the adult rearing methods and adult maintenance unit are recommended for Ae. albopictus mass rearing in support of the establishment of a medium-sized mosquito factory.

  10. Microencapsulated equine mesenchymal stromal cells promote cutaneous wound healing in vitro.

    PubMed

    Bussche, Leen; Harman, Rebecca M; Syracuse, Bethany A; Plante, Eric L; Lu, Yen-Chun; Curtis, Theresa M; Ma, Minglin; Van de Walle, Gerlinde R

    2015-04-11

    The prevalence of impaired cutaneous wound healing is high and treatment is difficult and often ineffective, leading to negative social and economic impacts for our society. Innovative treatments to improve cutaneous wound healing by promoting complete tissue regeneration are therefore urgently needed. Mesenchymal stromal cells (MSCs) have been reported to provide paracrine signals that promote wound healing, but (i) how they exert their effects on target cells is unclear and (ii) a suitable delivery system to supply these MSC-derived secreted factors in a controlled and safe way is unavailable. The present study was designed to provide answers to these questions by using the horse as a translational model. Specifically, we aimed to (i) evaluate the in vitro effects of equine MSC-derived conditioned medium (CM), containing all factors secreted by MSCs, on equine dermal fibroblasts, a cell type critical for successful wound healing, and (ii) explore the potential of microencapsulated equine MSCs to deliver CM to wounded cells in vitro. MSCs were isolated from the peripheral blood of healthy horses. Equine dermal fibroblasts from the NBL-6 (horse dermal fibroblast cell) line were wounded in vitro, and cell migration and expression levels of genes involved in wound healing were evaluated after treatment with MSC-CM or NBL-6-CM. These assays were repeated by using the CM collected from MSCs encapsulated in core-shell hydrogel microcapsules. Our salient findings were that equine MSC-derived CM stimulated the migration of equine dermal fibroblasts and increased their expression level of genes that positively contribute to wound healing. In addition, we found that equine MSCs packaged in core-shell hydrogel microcapsules had similar effects on equine dermal fibroblast migration and gene expression, indicating that microencapsulation of MSCs does not interfere with the release of bioactive factors. Our results demonstrate that the use of CM from MSCs might be a promising new therapy for impaired cutaneous wounds and that encapsulation may be a suitable way to effectively deliver CM to wounded cells in vivo.

  11. Influence of Mesenchymal Stem Cells Conditioned Media on Proliferation of Urinary Tract Cancer Cell Lines and Their Sensitivity to Ciprofloxacin.

    PubMed

    Maj, Malgorzata; Bajek, Anna; Nalejska, Ewelina; Porowinska, Dorota; Kloskowski, Tomasz; Gackowska, Lidia; Drewa, Tomasz

    2017-06-01

    Mesenchymal stem cells (MSCs) are known to interact with cancer cells through direct cell-to-cell contact and secretion of paracrine factors, although their exact influence on tumor progression in vivo remains unclear. To better understand how fetal and adult stem cells affect tumors, we analyzed viability of human renal (786-0) and bladder (T24) carcinoma cell lines cultured in conditioned media harvested from amniotic fluid-derived stem cells (AFSCs) and adipose-derived stem cells (ASCs). Both media reduced metabolic activity of 786-0 cells, however, decreased viability of T24 cells was noted only after incubation with conditioned medium from ASCs. To test the hypothesis that MSCs-secreted factors might be involved in chemoresistance acquisition, we further analyzed influence of mesenchymal stem cell conditioned media (MSC-CM) on cancer cells sensitivity to ciprofloxacin, that is considered as potential candidate agent for urinary tract cancers treatment. Significantly increased resistance to tested drug indicates that MSCs may protect cancer cells from chemotherapy. J. Cell. Biochem. 118: 1361-1368, 2017. © 2016 Wiley Periodicals, Inc. © 2016 Wiley Periodicals, Inc.

  12. Role of Oxidative Stress in the Suppression of Immune Responses in Peripheral Blood Mononuclear Cells Exposed to Combustible Tobacco Product Preparation.

    PubMed

    Arimilli, Subhashini; Schmidt, Eckhardt; Damratoski, Brad E; Prasad, G L

    2017-10-01

    Cigarette smoking is a major risk factor for several human diseases. Chronic inflammation, resulting from increased oxidative stress, has been suggested as a mechanism that contributes to the increased susceptibility of smokers to cancer and microbial infections. We have previously shown that whole-smoke conditioned medium (WS-CM) and total particulate matter (TPM) prepared from Kentucky 3R4F reference cigarettes [collectively called as combustible tobacco product preparations (TPPs)] potently suppressed agonist-stimulated cytokine secretion and target cell killing in peripheral blood mononuclear cells (PBMCs). Here we have investigated the role of oxidative stress from TPPs, which alters inflammatory responses in vitro. Particularly, we investigated the mechanisms of WS-CM-induced suppression of select cytokine secretions in Toll-like receptor (TLR) agonist-stimulated cells and target cell killing by effector cells in PBMCs. Pretreatment with N-acetyl cysteine (NAC), a precursor of reduced glutathione and an established anti-oxidant, protected against DNA damage and cytotoxicity caused by exposure to WS-CM. Similarly, secretion of tumor necrosis factor (TNF), interleukin (IL)-6, and IL-8 in response to TLR-4 stimulation was restored by pretreatment with NAC. Target cell killing, a functional measure of cytolytic cells in PBMCs, is suppressed by WS-CM. Pretreatment with NAC restored the target cell killing in WS-CM treated PBMCs. This was accompanied by higher perforin levels in the effector cell populations. Collectively, these data suggest that reducing oxidative stress caused by cigarette smoke components restores select immune responses in this ex vivo model.

  13. Electrocoagulation of simulated reactive dyebath effluent with aluminum and stainless steel electrodes.

    PubMed

    Arslan-Alaton, Idil; Kabdaşli, Işik; Vardar, Burcu; Tünay, Olcay

    2009-05-30

    Reactive dyebath effluents are ideal candidates for electrocoagulation due to their intensive color, medium strength, recalcitrant COD and high electrolyte (NaCl) content. The present study focused on the treatability of simulated reactive dyebath effluent (COD(o)=300 mg/L; color in terms of absorbance values A(o,436)=0.532 cm(-1), A(o,525)=0.693 cm(-1) and A(o,620)=0.808 cm(-1)) employing electrocoagulation with aluminum and stainless steel electrodes. Optimization of critical operating parameters such as initial pH (pH(o) 3-11), applied current density (J(c)=22-87 mA/cm(2)) and electrolyte type (NaCl or Na(2)SO(4)) improved the overall treatment efficiencies resulting in effective decolorization (99% using stainless steel electrodes after 60 min, 95% using aluminum electrodes after 90 min electrocoagulation) and COD abatement (93% with stainless steel electrodes after 60 min, 86% with aluminum electrodes after 90 min of reaction time). Optimum electrocoagulation conditions were established as pH(o) 5 and J(c)=22 mA/cm(2) for both electrode materials. The COD and color removal efficiencies also depended on the electrolyte type. No in situ, surplus adsorbable organically bound halogens (AOX) formation associated with the use of NaCl as the electrolyte during electrocoagulation was detected. An economical evaluation was also carried out within the frame of the study. It was demonstrated that electrocoagulation of reactive dyebath effluent with aluminum and stainless steel electrodes was a considerably less electrical energy-intensive, alternative treatment method as compared with advanced chemical oxidation techniques.

  14. Supplementation of Farta sheep fed hay with graded levels of concentrate mix consisting of noug seed meal and rice bran.

    PubMed

    Asmare, Bimrew; Melaku, Solomon; Peters, Kurt J

    2010-10-01

    The study was carried out at Woreta, Ethiopia, to determine feed intake, digestibility, body weight (BW) change, and profitability of Farta sheep fed pasture hay alone or supplemented with graded levels of concentrate mix (CM) consisting of noug seed meal (NSM) and rice bran in 2:1 ratio. Twenty yearling intact male Farta sheep with BW of 16.9 +/- 1.68 kg (mean +/- SD) were used in randomized complete block design arranged into five blocks of four animals. The dietary treatments consisting of sole natural pasture hay (T1, control), hay +200 g of CM dry matter (DM) (T2, low), hay +300 g of CM DM (T3, medium), and hay +400 g of CM DM (T4, high) were randomly assigned to sheep within each block. Common salt and water were available to animals all the time. The supplements were offered twice daily in equal portions at 0800 and 1600 h. Supplementation with the CM increased (P < 0.001) DM, organic matter, crude protein, neutral detergent fiber, and acid detergent fiber intake and digestibility. The final BW and daily BW gains were higher (P < 0.001) for the supplemented compared to the control treatment. The study also revealed supplementation improved feed conversion efficiency and profitability. Among the supplemented treatments, the high level of supplementation resulted in better (P < 0.001) nutrient utilization, animal performance, and profitability. Thus, the high level of supplementation is recommended based on biological performance and profitability under conditions of this study.

  15. The GLC8 - A miniature low cost ring laser gyroscope

    NASA Astrophysics Data System (ADS)

    Godart, D.-F.; Peghaire, J.-P.

    SAGEM is enlarging its family of ring laser gyros (RLG) which already includes a triangular 32-cm path-length gyro and a square 16-cm path-length gyro, in order to meet the increasing demand for low cost, medium accuracy strap-down inertial measurement units for applications such as short- and medium-range tactical missiles as well as aided navigation systems for aircrafts and land vehicles. Based on the experience acquired in the past 13 years in the RLG field, and especially in mirror manufacturing, SAGEM developed the GLC8 which has a square 8-cm path length cavity, central piezoelectric dither. It incorporates two cathodes, a single anode, and is technologically designed to minimize production-costs while optimizing the performance to global device size ratio. This gyro is characterized by a bias and a scale-factor stability respectively better than 0.5 deg/h and 100 ppm (1 sigma), and has an operating lifetime compatible with the most demanding relevant applications and a high robustness to mechanical environments.

  16. Current status and future prospects of research and development operations in traditional and complementary and alternative medicine manufacturing small- and medium-sized enterprises: a 2014 company-based survey.

    PubMed

    Ahn, Miyoung; Park, Eun-Ji; Woo, Jong-Min

    2017-12-01

    Small- and medium-sized enterprises (SMEs) have played key roles in the economic growth and technical innovation of traditional and complementary and alternative medicine (T&CM). Research and development (R&D) are critical activities for industrial progress. This study aimed to characterize the current status of SME R&D activities and to explore manufacturers' perceptions of R&D expansion. Records of the distribution of T&CM SMEs and R&D resources detailed in the 2014 Statistics of Korea T&CM Industries survey, a previously conducted survey on the industrial status of the T&CM field, were reviewed. Data on the perceptions of R&D activities were investigated through a company-based survey covering 285 T&CM-manufacturing SMEs. Greater than 99% of the 13,636 T&CM manufacturers at the time of the study were SMEs employing less than 50 workers. Natural cosmetics manufacturing SMEs (NC SMEs) had the highest R&D expenditures. NC SMEs rely heavily on internal R&D operations, which may contribute to their strong need for R&D collaboration with public research institutions and expanded T&CM-promoted R&D programs. "Digestive system disorders" are the main target diseases for current herbal and dietary supplement manufacturing SMEs and herbal medicine manufacturing SMEs. These SMEs tend to view their own product-related business as a priority for future R&D investment. This study represents the first attempt to assess SME perceptions of R&D activities. The findings herein can inform the design of sustainable programs that support R&D by reducing the gaps between the perspectives of T&CM product makers and policymakers.

  17. Simulating the impact of X-ray heating during the cosmic dawn

    NASA Astrophysics Data System (ADS)

    Ross, Hannah E.; Dixon, Keri L.; Iliev, Ilian T.; Mellema, Garrelt

    2017-07-01

    Upcoming observations of the 21-cm signal from the epoch of reionization will soon provide the first direct detection of this era. This signal is influenced by many astrophysical effects, including long-range X-ray heating of the intergalactic gas. During the preceding cosmic dawn era, the impact of this heating on the 21-cm signal is particularly prominent, especially before spin temperature saturation. We present the largest volume (349 Mpc comoving = 244 h-1Mpc) full numerical radiative transfer simulations to date of this epoch which include the effects of helium and multifrequency heating, both with and without X-ray sources. We show that X-ray sources contribute significantly to early heating of the neutral intergalactic medium and, hence, to the corresponding 21-cm signal. The inclusion of hard, energetic radiation yields an earlier, extended transition from absorption to emission compared to the stellar-only case. The presence of X-ray sources decreases the absolute value of the mean 21-cm differential brightness temperature. These hard sources also significantly increase the 21-cm fluctuations compared to the common assumption of temperature saturation. The 21-cm differential brightness temperature power spectrum is initially boosted on large scales, before decreasing on all scales. Compared to the case of the cold, unheated intergalactic medium, the signal has lower rms fluctuations and increased non-Gaussianity, as measured by the skewness and kurtosis of the 21-cm probability distribution functions. Images of the 21-cm signal with resolution around 11 arcmin still show fluctuations well above the expected noise for deep integrations with the SKA1-Low, indicating that direct imaging of the X-ray heating epoch could be feasible.

  18. Suppression of Mg propagation into subsequent layers grown by MOCVD

    NASA Astrophysics Data System (ADS)

    Agarwal, Anchal; Tahhan, Maher; Mates, Tom; Keller, Stacia; Mishra, Umesh

    2017-01-01

    Low temperature (LT) flow modulation epitaxy (FME) or "pulsed" growth was successfully used to prevent magnesium from Metalorganic Chemical Vapor Deposition (MOCVD) grown p-GaN:Mg layers riding into subsequently deposited n-type layers. Mg concentration in the subsequent layers was lowered from ˜1 × 1018 cm-3 for a medium temperature growth at 950 °C to ˜1 × 1016 cm-3 for a low temperature growth at 700 °C via FME. The slope of the Mg concentration drop in the 700 °C FME sample was 20 nm/dec—the lowest ever demonstrated by MOCVD. For growth on Mg implanted GaN layers, the drop for a medium temperature regrowth at 950 °C was ˜10 nm/dec compared to >120 nm/dec for a high temperature regrowth at 1150 °C. This drop-rate obtained at 950 °C or lower was maintained even when the growth temperature in the following layers was raised to 1150 °C. A controlled silicon doping series using LT FME was also demonstrated with the lowest and highest achieved doping levels being 5 × 1016 cm-3 and 6 × 1019 cm-3, respectively.

  19. Production of oil palm empty fruit bunch compost for ornamental plant cultivation

    NASA Astrophysics Data System (ADS)

    Trisakti, B.; Mhardela, P.; Husaini, T.; Irvan; Daimon, H.

    2018-02-01

    The aim of this research was to produce the oil palm empty fruit bunch (EFB) compost for ornamental plant cultivation. EFB compost was produced by chopping fresh EFB into 1-3 cm pieces, inserting the pieces into basket composter (33 cm W × 28 cm L × 40 cm H), and adding activated liquid organic fertilizer (ALOF) until moisture content (MC) in the range of 55-65%. During composting, the compost pile was turned every 3 days and the MC was maintained at 55-65% range by adding the ALOF. The compost processed was then mixed with sand and rice husk with a ratio of 1:1:1; 1:3:1; 1:0:1 and was used as a potting medium for planting some valuable ornamental plants i.e. cactus (cactaceae), sansevieria, and anthurium. Composting was carried out for 40 days and the compost characteristic were pH 9.0; MC 52.59%; WHC 76%; CN ratio 12.15; N 1.96%; P 0.58%; and K 0.95%. The compost-sand-husk rice mixture can be used as a growing medium where the best ratio for cactus, sansevieria, and anthurium was 1:3:1; 1:1:1; and 1:0:1, respectively.

  20. Primary stability, insertion torque, and bone density of conical implants with internal hexagon: is there a relationship?

    PubMed

    Trisi, Paolo; Berardi, Davide; Paolantonio, Michele; Spoto, Giuseppe; D'Addona, Antonio; Perfetti, Giorgio

    2013-05-01

    Between implants and peri-implant bone, there should be a minimum gap, without micromotions over a threshold, which could cause resorption and fibrosis. The higher the implant insertion torque, the higher will be the initial stability. The aim was to evaluate in vitro the correlation between micromotions and insertion torque of implants in bone of different densities. The test was performed on bovine bone of hard, medium, and soft density: 150 implants were used, 10 for each torque (20, 35, 45, 70, and 100 N/cm). Samples were fixed on a loading device. On each sample, we applied a 25-N horizontal force. Insertion torque and micromotions are statistically correlated. In soft bone with an insertion force of 20 and 35 N/cm, the micromotion resulted significantly over the risk threshold, which was not found with an insertion force of 45 and 70 N/cm and in hard and medium bones with any insertion torque. The increase in insertion torque reduces the amount of micromotions between implant and bone. Therefore, the immediate loading may be considered a valid therapeutic choice, even in low-density bone, as long as at least 45 N/cm of insertion torque is reached.

  1. Dynamics of supernova driven superbubbles

    NASA Astrophysics Data System (ADS)

    Yadav, Naveen; Mukherjee, Dipanjan; Sharma, Prateek; Nath, Biman

    2015-08-01

    Energy injection by supernovae is believed to be one of the primary sources which powers the expansion of supershells. There is a qualitative difference between isolated supernovae (SNe) and overlapping SNe. For typical interstellar medium (ISM) conditions an isolated supernova loses most of the injected mechanical energy by 1 Myr. In contrast, for SNe going off in bubbles the radiative losses are much smaller. While the outer shock going off in the dense ISM (~1 cm-3) becomes radiative well before 1 Myr, there is a strong non-radiative termination shock that keeps the bubble over-pressured till the lifetime of the OB association (10s of Myr; Sharma et al. 2014). This has relevance for supernova feedback in galaxy formation simulations. In our previous 1-D treatment all the SNe were assumed to occur at the same location in space. It was found that a steady wind inside the bubble (Chevalier & Clegg 1985) can occur only if the number of SNe is large (>~104) and a supernova going off inside the bubble can thermalize within the termination shock. In the present work we study the effect of SNe separated in both space and time using 3-D hydrodynamic simulations with radiative cooling. If the separation between SNe is larger than the remnant’s radius at the time it becomes radiative, SNe are in the isolated regime. The explosion energy is deposited as thermal energy in a uniform, static interstellar medium (ISM) with temperature 104 K, corresponding to the warm neutral medium. The key parameters of our idealized setup are the ISM density (ngas), the number of SNe (N★) and the spatial separation between SNe (Rcl). The shock radius when it becomes radiative depends on the ISM density and number of SNe. We obtain the critical values of the key parameters (ngas, N★, Rcl) which lead to the formation of a superbubble. e.g., at least 103 SNe are required to maintain an over-pressured bubble at 20 Myr in an ISM with 1 cm-3 similarly 102 SNe going off within a region of 100 pc in an ISM with 1 cm-3 are effectively in the isolated regime. We also compare our simulation results with observational data on superbubbles.

  2. Comparative study on toxicity of ZnO and TiO2 nanoparticles on Artemia salina: effect of pre-UV-A and visible light irradiation.

    PubMed

    Bhuvaneshwari, M; Sagar, Bhawana; Doshi, Siddharth; Chandrasekaran, N; Mukherjee, Amitava

    2017-02-01

    This study evaluated the toxicity potential of ZnO and TiO 2 nanoparticles under pre-UV-A irradiation and visible light condition on Artemia salina. The nanoparticle suspension was prepared in seawater medium and exposed under pre-UV-A (0.23 mW/cm 2 ) and visible light (0.18 mW/cm 2 ) conditions. The aggregation profiles of both nanoparticles (NPs) and dissolution of ZnO NPs under both irradiation conditions at various kinetic intervals (1, 24, 48 h) were studied. The 48-h LC 50 values were found to be 27.62 and 71.63 mg/L for ZnO NPs and 117 and 120.9 mg/L for TiO 2 NPs under pre-UV-A and visible light conditions. ZnO NPs were found to be more toxic to A. salina as compared to TiO 2 NPs. The enhanced toxicity was observed under pre-UV-A-irradiated ZnO NPs, signifying its phototoxicity. Accumulation of ZnO and TiO 2 NPs into A. salina depends on the concentration of particles and type irradiations. Elimination of accumulated nanoparticles was also evident under both irradiation conditions. Other than ZnO NPs, the dissolved Zn 2+ also had a significant effect on toxicity and accumulation in A. salina. Increased catalase (CAT) activity in A. salina indicates the generation of oxidative stress due to NP interaction. Thus, this study provides an understanding of the toxicity of photoreactive ZnO and TiO 2 NPs as related to the effects of pre-UV-A and visible light irradiation.

  3. Raman spectroscopy analysis of differences in composition of spent culture media of in vitro cultured preimplantation embryos isolated from normal and fat mice dams.

    PubMed

    Fabian, Dušan; Kačmarová, Martina; Kubandová, Janka; Čikoš, Štefan; Koppel, Juraj

    2016-06-01

    The aim of the present study was to compare overall patterns of metabolic activity of in vitro cultured preimplantation embryos isolated from normal and fat mice dams by means of non-invasive profiling of spent culture media using Raman spectroscopy. To produce females with two different types of body condition (normal and fat), a previously established two-generation model was used, based on overfeeding of experimental mice during prenatal and early postnatal development. Embryos were isolated from spontaneously ovulating and naturally fertilized dams at the 2-cell stage of development and cultured to the blastocyst stage in synthetic oviductal medium KSOMaa. Embryos from fat mice (displaying significantly elevated body weight and fat) showed similar developmental capabilities in vitro as embryos isolated from normal control dams (displaying physiological body weight and fat). The results show that alterations in the composition of culture medium caused by the presence of developing mouse preimplantation embryos can be detected using Raman spectroscopy. Metabolic activity of embryos was reflected in evident changes in numerous band intensities in the 1620-1690cm(-1) (amide I) region and in the 1020-1140cm(-1) region of the Raman spectrum for KSOMaa. Moreover, multivariate analysis of spectral data proved that the composition of proteins and other organic compounds in spent samples obtained after the culture of embryos isolated from fat dams was different from that in spent samples obtained after the culture of embryos from control dams. This study demonstrates that metabolic activity of cultured preimplantation embryos might depend on the body condition of their donors. Copyright © 2016 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.

  4. Effects of intratracheal mesenchymal stromal cell therapy during recovery and resolution after ventilator-induced lung injury.

    PubMed

    Curley, Gerard F; Ansari, Bilal; Hayes, Mairead; Devaney, James; Masterson, Claire; Ryan, Aideen; Barry, Frank; O'Brien, Timothy; Toole, Daniel O'; Laffey, John G

    2013-04-01

    Mesenchymal stromal cells (MSCs) have been demonstrated to attenuate acute lung injury when delivered by intravenous or intratracheal routes. The authors aimed to determine the efficacy of and mechanism of action of intratracheal MSC therapy and to compare their efficacy in enhancing lung repair after ventilation-induced lung injury with intravenous MSC therapy. : After induction of anesthesia, rats were orotracheally intubated and subjected to ventilation-induced lung injury (respiratory rate 18(-1) min, P insp 35 cm H2O,) to produce severe lung injury. After recovery, animals were randomized to receive: (1) no therapy, n = 4; (2) intratracheal vehicle (phosphate-buffered saline, 300 µl, n = 8); (3) intratracheal fibroblasts (4 × 10 cells, n = 8); (4) intratracheal MSCs (4 × 10(6) cells, n = 8); (5) intratracheal conditioned medium (300 µl, n = 8); or (6) intravenous MSCs (4 × 10(6) cells, n = 4). The extent of recovery after acute lung injury and the inflammatory response was assessed after 48 h. Intratracheal MSC therapy enhanced repair after ventilation-induced lung injury, improving arterial oxygenation (mean ± SD, 146 ± 3.9 vs. 110.8 ± 21.5 mmHg), restoring lung compliance (1.04 ± 0.11 vs. 0.83 ± 0.06 ml · cm H2O(-1)), reducing total lung water, and decreasing lung inflammation and histologic injury compared with control. Intratracheal MSC therapy attenuated alveolar tumor necrosis factor-α (130 ± 43 vs. 488 ± 211 pg · ml(-1)) and interleukin-6 concentrations (138 ± 18 vs. 260 ± 82 pg · ml(-1)). The efficacy of intratracheal MSCs was comparable with intravenous MSC therapy. Intratracheal MSCs seemed to act via a paracine mechanism, with conditioned MSC medium also enhancing lung repair after injury. Intratracheal MSC therapy enhanced recovery after ventilation-induced lung injury via a paracrine mechanism, and was as effective as intravenous MSC therapy.

  5. Methods to Evaluate Cytotoxicity and Immunosuppression of Combustible Tobacco Product Preparations

    PubMed Central

    Arimilli, Subhashini; Damratoski, Brad E.; G.L., Prasad

    2015-01-01

    Among other pathophysiological changes, chronic exposure to cigarette smoke causes inflammation and immune suppression, which have been linked to increased susceptibility of smokers to microbial infections and tumor incidence. Ex vivo suppression of receptor-mediated immune responses in human peripheral blood mononuclear cells (PBMCs) treated with smoke constituents is an attractive approach to study mechanisms and evaluate the likely long-term effects of exposure to tobacco products. Here, we optimized methods to perform ex vivo assays using PBMCs stimulated by bacterial lipopolysaccharide, a Toll-like receptor-4 ligand. The effects of whole smoke-conditioned medium (WS-CM), a combustible tobacco product preparation (TPP), and nicotine were investigated on cytokine secretion and target cell killing by PBMCs in the ex vivo assays. We show that secreted cytokines IFN-γ, TNF, IL-10, IL-6, and IL-8 and intracellular cytokines IFN-γ, TNF-α, and MIP-1α were suppressed in WS-CM-exposed PBMCs. The cytolytic function of effector PBMCs, as determined by a K562 target cell killing assay was also reduced by exposure to WS-CM; nicotine was minimally effective in these assays. In summary, we present a set of improved assays to evaluate the effects of TPPs in ex vivo assays, and these methods could be readily adapted for testing other products of interest. PMID:25650834

  6. Methods to evaluate cytotoxicity and immunosuppression of combustible tobacco product preparations.

    PubMed

    Arimilli, Subhashini; Damratoski, Brad E; G L, Prasad

    2015-01-10

    Among other pathophysiological changes, chronic exposure to cigarette smoke causes inflammation and immune suppression, which have been linked to increased susceptibility of smokers to microbial infections and tumor incidence. Ex vivo suppression of receptor-mediated immune responses in human peripheral blood mononuclear cells (PBMCs) treated with smoke constituents is an attractive approach to study mechanisms and evaluate the likely long-term effects of exposure to tobacco products. Here, we optimized methods to perform ex vivo assays using PBMCs stimulated by bacterial lipopolysaccharide, a Toll-like receptor-4 ligand. The effects of whole smoke-conditioned medium (WS-CM), a combustible tobacco product preparation (TPP), and nicotine were investigated on cytokine secretion and target cell killing by PBMCs in the ex vivo assays. We show that secreted cytokines IFN-γ, TNF, IL-10, IL-6, and IL-8 and intracellular cytokines IFN-γ, TNF-α, and MIP-1α were suppressed in WS-CM-exposed PBMCs. The cytolytic function of effector PBMCs, as determined by a K562 target cell killing assay was also reduced by exposure to WS-CM; nicotine was minimally effective in these assays. In summary, we present a set of improved assays to evaluate the effects of TPPs in ex vivo assays, and these methods could be readily adapted for testing other products of interest.

  7. LPS-inducible factor(s) from activated macrophages mediates cytolysis of Naegleria fowleri amoebae

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

    Cleary, S.F.; Marciano-Cabral, F.

    1986-03-01

    Soluble cytolytic factors of macrophage origin have previously been described with respect to their tumoricidal activity. The purpose of this study was to investigate the mechanism and possible factor(s) responsible for cytolysis of the amoeba Naegleria fowleri by activated peritoneal macrophages from B6C3F1 mice. Macrophages or conditioned medium (CM) from macrophage cultures were incubated with /sup 3/H-Uridine labeled amoebae. Percent specific release of label served as an index of cytolysis. Bacille Calmette-Guerin (BCG) and Corynebacterium parvum macrophages demonstrated significant cytolysis of amoebae at 24 h with an effector to target ratio of 10:1. Treatment of macrophages with inhibitors of RNAmore » or protein synthesis blocked amoebicidal activity. Interposition of a 1 ..mu..m pore membrane between macrophages and amoebae inhibited killing. Inhibition in the presence of the membrane was overcome by stimulating the macrophages with LPS. CM from SPS-stimulated, but not unstimulated, cultures of activated macrophages was cytotoxic for amoebae. The activity was heat sensitive and was recovered from ammonium sulfate precipitation of the CM. Results indicate that amoebicidal activity is mediated by a protein(s) of macrophage origin induced by target cell contact or stimulation with LPS.« less

  8. Fabrication and Compressive Properties of Low to Medium Porosity Closed-Cell Porous Aluminum Using PMMA Space Holder Technique

    PubMed Central

    Jamal, Nur Ayuni; Tan, Ai Wen; Yusof, Farazila; Katsuyoshi, Kondoh; Hisashi, Imai; Singh, S.; Anuar, Hazleen

    2016-01-01

    In recent years, closed-cell porous Aluminum (Al) has drawn increasing attention, particularly in the applications requiring reduced weight and energy absorption capability such as in the automotive and aerospace industries. In the present work, porous Al with closed-cell structure was successfully fabricated by powder metallurgy technique using PMMA as a space holder. The effects of the amount of PMMA powder on the porosity, density, microstructure and compressive behaviors of the porous specimens were systematically evaluated. The results showed that closed-cell porous Al having different porosities (12%–32%) and densities (1.6478 g/cm3, 1.5125 g/cm3 and 1.305 g/cm3) could be produced by varying the amount of PMMA (20–30 wt %). Meanwhile, the compressive behavior results demonstrated that the plateau stress decreased and the energy absorption capacity increased with increasing amount of PMMA. However, the maximum energy absorption capacity was achieved in the closed-cell porous Al with the addition of 25 wt % PMMA. Therefore, fabrication of closed-cell porous Al using 25 wt % PMMA is considered as the optimal condition in the present study since the resultant closed-cell porous Al possessed good combinations of porosity, density and plateau stress, as well as energy absorption capacity. PMID:28773377

  9. Investigation of mitomycin-C-treated fibroblasts in 3-D collagen gel and conditioned medium for keratinocyte proliferation.

    PubMed

    Huang, Yi-Chau; Wang, Tzu-Wei; Sun, Jui-Sheng; Lin, Feng-Huei

    2006-03-01

    Fibroblasts produce a spectrum of necessary growth factors essential for growth and proliferation of a variety of cell types. In this study, the paracrine effect of mitomycin-C-treated fibroblasts with various densities in collagen gel for keratinocyte proliferation was investigated from which an optimum cell density and optimum conditioned medium would be determined to expand keratinocyte without further differentiation for skin equivalent tissue engineering. The optimum cell density in collagen feeder gel for optimum collected medium preparation will be determined by checking the level of keratinocyte growth factor and granulocyte macrophage colony-stimulating factor in conventional medium. The results showed that the cell density of 1 x 10(5) cells/gel in the feeder gel is better to produce optimum collected medium. The conditioned medium is prepared by mixing together the optimum collected medium and molecular cellular and developmental biology (MCDB) 153 medium in different ratios for keratinocyte growth. The keratinocyte viability will be measured by 3-(4,5-dimethyl-thiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay to determine the optimum conditioned medium. From the study, 67% conditioned medium was supposed as the better medium for keratinocyte proliferation. In this experiment, the optimum cell density in feeder gel to coculture with keratinocytes is also determined as 1 x 10(5) cells/gel. Keratin 10 (K10) and Terminal Deoxynucleotidyl Transferase Mediated dUTP Nick End Labeling stain will be used to check the cell differentiation and apoptosis, respectively. The results suggest that keratinocytes should not be cultured in postconfluent conditions due to undesired apoptosis and differentiation. The result of cell viability from passages to passages shows that the optimum feeder gel plays a more important role to the keratinocyte proliferation than that of optimum conditioned medium. Keratinocytes cultured with optimum feeder gel in 67% conditioned medium could effectively promote proliferation, inhibit apoptosis, and prevent differentiation. The combination of conditioned media and feeder gel to culture keratinocytes without external supplements can provide an inexpensive way for keratinocyte proliferation and construct an environment for real-time communication between the two cells. The results conclude that keratinocyte cultivation in feeder gel with modified medium should be feasible in the production of high quality keratinocytes for skin equivalents preparation.

  10. Multiple shoot induction from axillary bud cultures of the medicinal orchid, Dendrobium longicornu

    PubMed Central

    Dohling, Stadwelson; Kumaria, Suman; Tandon, Pramod

    2012-01-01

    Background and aims Dendrobium longicornu, commonly known as the ‘Long-horned Dendrobium’, is an endangered and medicinally important epiphytic orchid. Over-exploitation and habitat destruction seriously threaten this orchid in Northeast India. Our objective was to develop an efficient protocol for the mass propagation of D. longicornu using axillary bud segments. Methodology and principal results Axillary buds cultured in Murashige and Skoog semi-solid medium supplemented with α-naphthalene acetic acid (NAA), 2,4-dichlorophenoxy acetic acid (2,4-D) and 6-benzylaminopurine (BAP) readily developed into plantlets. These formed either directly from shoot buds or from intermediary protocorm-like bodies (PLBs). The maximum explant response (86.6 %) was obtained in medium supplemented with NAA at 30 µM, while the maximum number of shoots (4.42) and maximum bud-forming capacity (3.51) were observed in medium containing 15 µM BAP and 5 µM NAA in combination. Protocorm-like bodies were obtained when the medium contained 2,4-D. The maximum number of explants forming PLBs (41.48 %) was obtained in medium containing 15 µM BAP and 15 µM 2,4-D. Well-developed plantlets obtained after 20–25 weeks of culture were acclimatized and eventually transferred to the greenhouse. Over 60 % of these survived to form plants ∼3–4 cm tall after 90 days in glasshouse conditions using a substrate of crushed brick and charcoal, shredded bark and moss. Conclusions The method described can readily be used for the rapid and large-scale regeneration of D. longicornu. Its commercial adoption would reduce the collection of this medicinally important and increasingly rare orchid from the wild. PMID:23136638

  11. Optical measurements of absorption changes in two-layered diffusive media

    NASA Astrophysics Data System (ADS)

    Fabbri, Francesco; Sassaroli, Angelo; Henry, Michael E.; Fantini, Sergio

    2004-04-01

    We have used Monte Carlo simulations for a two-layered diffusive medium to investigate the effect of a superficial layer on the measurement of absorption variations from optical diffuse reflectance data processed by using: (a) a multidistance, frequency-domain method based on diffusion theory for a semi-infinite homogeneous medium; (b) a differential-pathlength-factor method based on a modified Lambert-Beer law for a homogeneous medium and (c) a two-distance, partial-pathlength method based on a modified Lambert-Beer law for a two-layered medium. Methods (a) and (b) lead to a single value for the absorption variation, whereas method (c) yields absorption variations for each layer. In the simulations, the optical coefficients of the medium were representative of those of biological tissue in the near-infrared. The thickness of the first layer was in the range 0.3-1.4 cm, and the source-detector distances were in the range 1-5 cm, which is typical of near-infrared diffuse reflectance measurements in tissue. The simulations have shown that (1) method (a) is mostly sensitive to absorption changes in the underlying layer, provided that the thickness of the superficial layer is ~0.6 cm or less; (2) method (b) is significantly affected by absorption changes in the superficial layer and (3) method (c) yields the absorption changes for both layers with a relatively good accuracy of ~4% for the superficial layer and ~10% for the underlying layer (provided that the absorption changes are less than 20-30% of the baseline value). We have applied all three methods of data analysis to near-infrared data collected on the forehead of a human subject during electroconvulsive therapy. Our results suggest that the multidistance method (a) and the two-distance partial-pathlength method (c) may better decouple the contributions to the optical signals that originate in deeper tissue (brain) from those that originate in more superficial tissue layers.

  12. PROBING X-RAY ABSORPTION AND OPTICAL EXTINCTION IN THE INTERSTELLAR MEDIUM USING CHANDRA OBSERVATIONS OF SUPERNOVA REMNANTS

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

    Foight, Dillon R.; Slane, Patrick O.; Güver, Tolga

    We present a comprehensive study of interstellar X-ray extinction using the extensive Chandra supernova remnant (SNR) archive and use our results to refine the empirical relation between the hydrogen column density and optical extinction. In our analysis, we make use of the large, uniform data sample to assess various systematic uncertainties in the measurement of the interstellar X-ray absorption. Specifically, we address systematic uncertainties that originate from (i) the emission models used to fit SNR spectra; (ii) the spatial variations within individual remnants; (iii) the physical conditions of the remnant such as composition, temperature, and non-equilibrium regions; and (iv) themore » model used for the absorption of X-rays in the interstellar medium. Using a Bayesian framework to quantify these systematic uncertainties, and combining the resulting hydrogen column density measurements with the measurements of optical extinction toward the same remnants, we find the empirical relation N {sub H} = (2.87 ± 0.12) × 10{sup 21} A {sub V} cm{sup 2}, which is significantly higher than the previous measurements.« less

  13. Effect of cytokinins on in vitro multiplication of Sophora tonkinensis

    PubMed Central

    Jana, Sonali; Sivanesan, Iyyakkannu; Jeong, Byoung Ryong

    2013-01-01

    Objective To determine the effects of different cytokinins at various concentrations on in vitro shoot multiplication of an important medicinal plant. Methods Nodal explants (1.5-2.0 cm) of Sophora tonkinensis were used. Multiple shoots were induced from nodal explants cultured on the Murashige and Skoog (MS) medium supplemented with 0.0, 0.5, 1.0, 2.0, 4.0, 8.0, or 16.0 µmol 2-isopentyladenine (2iP), N6 benzyladenine, kinetin or thiadiazuron. Results Among the four investigated cytokinins, 2iP showed the best response for shoot multiplication. Maximum shoot induction (75%) was achieved on the MS medium supplemented with 2.0 µmol 2iP, with a mean number of 5.0 shoots per explant. In comparison to other cytokinins tried, 2iP showed the highest shoot elongation with a mean shoot length of 4.8 cm. Root initiation was observed within 15 d within the transfer of shoots onto the MS basal medium, and the rooting percentage was 100% with a mean number of 5.4 roots per shoot and root length of 6.2 cm over a period of 4 weeks. The healthy plants, hardened and transferred to a greenhouse for proper acclimatization, exhibited 100% survival. Conclusions It can be summarized that 2iP is the optimal plant growth regulator for Sophora multiplication. PMID:23836310

  14. Floating cultivation of marine cyanobacteria using coal fly ash.

    PubMed

    Matsumoto, M; Yoshida, E; Takeyama, H; Matsunaga, T

    2000-01-01

    The aim of this study was to develop improved methodologies for bulk culturing of biotechnologically useful marine cyanobacteria in the open ocean. We have investigated the viability of using coal fly ash (CFA) blocks as the support medium in a novel floating culture system for marine micro-algae. The marine cyanobacterium Synechococcus sp. NKBG 040607 was found to adhere to floating CFA blocks in liquid culture medium. Maximum density of attached cells of 2.0 x 10(8) cells/cm2 was achieved using seawater. The marine cyanobacterium Synechococcus sp. NKBG 042902 weakly adhered to floating CFA blocks in BG-11 medium. Increasing the concentration of calcium ion in the culture medium enhanced adherence to CFA blocks.

  15. Sedimentary characteristics of the 2004 Indian Ocean tsunami in Ban Talae Nok, southwestern Ranong province, Thailand

    NASA Astrophysics Data System (ADS)

    Monecke, K.; Beitel, J.; Moran, K.; Moore, A.

    2006-12-01

    Ban Talae Nok, a village on the Andaman shoreline of Thailand, was hit by the December 26, 2004, tsunami with wave heights up to ~13 meters. Eyewitnesses reported the passage of four to five waves with the second being the largest, followed by the third and fourth waves. The tsunami flooded an area with open grassy fields, small cashew nut plantations and a wetland within a local swale. The wave stopped against hills ~500 m from the shoreline, where watermarks still indicate a flow depth of approximately 1 m. Erosion at the beach is marked by a ~30 cm high scarp cutting a former gravelly beach trail ~60 m inshore of the present shoreline. Deposition of tsunami sand started behind the former beach trail at 80 m inshore. The tsunami deposit changes significantly in thickness and composition along a flow parallel transect that was measured and sampled within this study. The most seaward deposit is about 10 cm thick and consists of three distinct layers that show internal as well as overall normal grading from coarse sand into fine sand. The coarse base contains gravels from the old beach trail and shell fragments. Locally, cross stratification is visible at the top. Farther landward the deposit thins and only one normally graded layer is visible. Behind a small ridge where the wetland begins, the tsunami sediments again reveal three normally graded layers with shell- rich, medium-coarse sand grading into brown-gray mud, probably eroded from the wetland. This deposit thins farther inland from ~30 cm to 8 cm and consists of only one layer. The thickest deposit along the transect is 125 cm thick and can be found in a low at the landward end of the wetland. It consists of normally graded coarse to fine sand with rip-up clasts at the base and climbing ripples in the middle of the deposit section. In the adjacent grassy field the deposit is up to 40 cm thick and consists of medium to coarse sand with shell fragments grading into fine to medium sands, which continue to the foot of the hills.

  16. Effects of UV-B Radiation and Periodic Desiccation on the Morphogenesis of the Edible Terrestrial Cyanobacterium Nostoc flagelliforme

    PubMed Central

    Feng, Yan-Na; Zhang, Zhong-Chun; Feng, Jun-Li

    2012-01-01

    The terrestrial cyanobacterium Nostoc flagelliforme Berk. et M. A. Curtis has been a popular food and herbal ingredient for hundreds of years. To meet great market demand and protect the local ecosystem, for decades researchers have tried to cultivate N. flagelliforme but have failed to get macroscopic filamentous thalli. In this study, single trichomes with 50 to 200 vegetative cells were induced from free-living cells by low light and used to investigate the morphogenesis of N. flagelliforme under low UV-B radiation and periodic desiccation. Low-fluence-rate UV-B (0.1 W m−2) did not inhibit trichome growth; however, it significantly increased the synthesis of extracellular polysaccharides and mycosporine-like amino acids and promoted sheath formation outside the trichomes. Under low UV-B radiation, single trichomes developed into filamentous thalli more than 1 cm long after 28 days of cultivation, most of which grew separately in liquid BG11 medium. With periodic desiccation treatment, the single trichomes formed flat or banded thalli that grew up to 2 cm long after 3 months on solid BG11 medium. When trichomes were cultivated on solid BG11 medium with alternate treatments of low UV-B and periodic desiccation, dark and scraggly filamentous thalli that grew up to about 3 cm in length after 40 days were obtained. In addition, the cultivation of trichomes on nitrogen-deficient solid BG11 medium (BG110) suggested that nitrogen availability could affect the color and lubricity of newly developed thalli. This study provides promising techniques for artificial cultivation of N. flagelliforme in the future. PMID:22865081

  17. Measurement of hydraulic characteristics of porous media used to grow plants in microgravity

    NASA Technical Reports Server (NTRS)

    Steinberg, Susan L.; Poritz, Darwin

    2005-01-01

    Understanding the effect of gravity on hydraulic properties of plant growth medium is essential for growing plants in space. The suitability of existing models to simulate hydraulic properties of porous medium is uncertain due to limited understanding of fundamental mechanisms controlling water and air transport in microgravity. The objective of this research was to characterize saturated and unsaturated hydraulic conductivity (K) of two particle-size distributions of baked ceramic aggregate using direct measurement techniques compatible with microgravity. Steady state (Method A) and instantaneous profile measurement (Method B) methods for K were used in a single experimental unit with horizontal flow through thin sections of porous medium providing an earth-based analog to microgravity. Comparison between methods was conducted using a crossover experimental design compatible with limited resources of space flight. Satiated (natural saturation) K ranged from 0.09 to 0.12 cm s-1 and 0.5 to >1 cm s-1 for 0.25- to 1- and 1- to 2-mm media, respectively. The K at the interaggregate/intraaggregate transition was approximately 10(-4) cm s-1 for both particle-size distributions. Significant differences in log(10)K due to method and porous medium were less than one order of magnitude and were attributed to variability in air entrapment. The van Genuchten/Mualem parametric models provided an adequate prediction of K of the interaggregate pore space, using residual water content for that pore space. The instantaneous profile method covers the range of water contents relevant to plant growth using fewer resources than Method A, all advantages for space flight where mass, volume, and astronaut time are limited.

  18. Desizing of Starch Containing Cotton Fabrics Using Near Atmospheric Pressure, Cold DC Plasma Treatment

    NASA Astrophysics Data System (ADS)

    Prasath, A.; Sivaram, S. S.; Vijay Anand, V. D.; Dhandapani, Saravanan

    2013-03-01

    An attempt has been made to desize the starch containing grey cotton fabrics using the DC plasma with oxygen as the gaseous medium. Process conditions of the plasma reactor were optimized in terms of distance between the plates (3.2 cm), applied voltage (600 V) and applied pressure (0.01 bar) to obtain maximum desizing efficiency. No discolouration was observed in the hot water extracts of the desized sample in presence of iodine though relatively higher solvent extractable impurities (4.53 %) were observed in the plasma desized samples compared to acid desized samples (3.38 %). Also, significant weight loss, improvements in plasma desized samples were observed than that of grey fabrics in terms of drop absorbency.

  19. [Ultraviolet spectrophotometric determination of tenoxicam using iodine solution as reagent].

    PubMed

    Mândrescu, Mariana; Spac, A F; Dorneanu, V

    2009-01-01

    For the tenoxicam determination (Nonsteroidal Antiinflammatory Drug - NSAID) it was developed a spectrophotometric method, after the coupling reaction of tenoxicam with iodine, in methanolic medium, with maximum of absorbance at 289 nm. The practical working conditions were established. In the 0.5 divided by 5.0 microg/mL range of tenoxicam concentration, were used 5 x 10(-3) M iodine solution and 0.5 N hydrocloric acid. The stability of product were evaluated for 30 minutes. The developed method was validated. The method showed a good linearity in the range of 0.5 divided 5.0 microg/mL (the correlation coefficient r = 0.9995). The detection limit (LD) was 0.14 microg/mL and the quantification limit (LQ) was 0.49 microg/mL. There were established the precison (RSD = 1.90%) and the accuracy-recovery in the range 97.27 divided by 102.56% with a mean recovery of 99.49%. The experimental results demonstrated a good sensibility. The specific absorptivity for this method is A1%(1 cm,289 nm) = 1770 much higher than tenoxicam in methanol (A1%(1 cm,360 nm) = 323).

  20. Effect of age on marrow macrophage number and function.

    PubMed

    Wang, C Q; Udupa, K B; Xiao, H; Lipschitz, D A

    1995-10-01

    Employing flow cytometry and a monoclonal antibody against the murine macrophage antigen, Mac-1, we found a significant increase in the number of marrow macrophages in aged mice. This was reflected as significant increase with age in the number of alpha-naphthyl acetate esterase positive cells, as well as in colony forming unit-macrophage (CFU-M) progenitor cells. Macrophages from the marrow of old mice generated significantly less tumor necrosis factor alpha (TNF alpha) than did macrophages from young mice, either spontaneously or when activated by granulocyte-macrophage colony stimulating factor (GM-CSF). Furthermore, conditioned medium (CM) derived from either marrow or peritoneal macrophages of old mice caused less suppression of burst forming unit-erythroid (BFU-E) colony growth than did CM obtained from young mice. Aging, therefore, is associated with an increase in the number of marrow macrophages that have an impaired ability to generate or release cytokines. The increase in macrophage number may reflect a compensation for their reduced function. Altered macrophage number and function may contribute to the age-related decline in hematopoietic reserve capacity.

  1. Involvement of Vascular Endothelial Growth Factor in Kaposi's Sarcoma Associated with Acquired Immunodeficiency Syndrome

    PubMed Central

    Sakurada, Shinsaku; Kato, Tetsuji; Mashiba, Kohichi; Mori, Shigeo

    1996-01-01

    To examine the role of vascular endothelial growth factor (VEGF) in the development of edema associated with Kaposi's sarcoma (KS) in acquired immunodeficiency syndrome (AIDS), we exploited animal model systems to detect the activity that induces vascular hyper‐permeability (VHP) using cultured AIDS‐KS spindle cells. Cultured AIDS‐KS spindle cells and conditioned medium (AIDS‐KS‐CM) that had been semi‐purified through a heparin affinity column were tested for the ability to induce VHP in animals. The AIDS‐KS spindle cells and AIDS‐KS‐CM induced VHP that was histamine‐independent. The VHP‐inducing activity was detected in the 0.5 M NaCl fraction from the heparin affinity column and was blocked by anti‐VEGF neutralizing antibody. In addition, the production of VEGF was demonstrated in fresh AIDS‐KS tissue as well as in cultured AIDS‐KS cells, while control cells were negative for VEGF production. From these observations, we concluded that AIDS‐KS cells produce a factor(s) that promotes VHP, and this factor could be VEGF. PMID:9045943

  2. A short peptide GPIGS promotes proliferation of hair bulb keratinocytes and accelerates hair regrowth in mice.

    PubMed

    Tsuruda, Akinori; Kawano, Yasuhiro; Maekawa, Takaaki; Oka, Syuichi

    2005-03-01

    The aim of this study was to discover a novel agent that promotes hair growth. We carried out a screening test in 298 types of conditioned medium (CM) from cultures of bacteria by using a hair bulb keratinocyte (HBK) growth assay. As a result, we found a HBK growth factor in the CM of Bacillus sp. M18. This HBK growth factor was purified by collecting biologically active fractions in three steps, including HP-20 batch processing, LH-20 chromatography and C18 reverse-phase high-pressure liquid chromatography, and identified as a short peptide GPIGS. GPIGS increased Akt phosphorylation in HBKs. Moreover, the GPIGS-stimulated HBK growth was inhibited by the treatment with LY294002, an inhibitor of phosphatidylinositol 3-kinase (PI-3K). These results suggest that GPIGS promotes HBK growth via the PI-3K/Akt pathway. In addition to in vitro tests, GPIGS was found to accelerate hair regrowth in telogen mice. Our results indicate that GPIGS is a potential agent to promote hair growth.

  3. Pressure shift coefficient measurements in an RF discharge for Ar 4s[3/2]2—5p[3/2]3 transition with the help of diodelaser absorption spectroscopy

    NASA Astrophysics Data System (ADS)

    Chernyshov, A. K.; Mikheyev, P. A.; Lunev, N. N.; Azyazov, V. N.

    2018-04-01

    Optically pumped all-rare-gas laser (OPRGL) with unique properties were recently proposed with a possibility to obtain the laser power on the order of hundreds of Watts from a cubic centimeter. To provide high laser efficiency, the pumping radiation has to match the absorption spectrum of the rare gas metastables. To meet this condition a reliable diagnostics of the key parameters of the active medium is required and knowledge of the broadening and shift coefficients for corresponding transitions of rare gases is necessary. In this paper, the diode-laser absorption spectroscopy was employed to determine the pressure shift coefficient for 811.5 nm Ar line. The value of obtained coefficient in pure argon reduced to 300 K is -(2.1 ± 0.1) × 10-10 s-1cm3. In the course of the study the pressure broadening coefficient was also evaluated and found to be (2.4 ± 0.5) × 10-10 s-1cm3.

  4. The relative impact of photoionizing radiation and stellar winds on different environments

    NASA Astrophysics Data System (ADS)

    Haid, S.; Walch, S.; Seifried, D.; Wünsch, R.; Dinnbier, F.; Naab, T.

    2018-05-01

    Photoionizing radiation and stellar winds from massive stars deposit energy and momentum into the interstellar medium (ISM). They might disperse the local ISM, change its turbulent multi-phase structure, and even regulate star formation. Ionizing radiation dominates the massive stars' energy output, but the relative effect of winds might change with stellar mass and the properties of the ambient ISM. We present simulations of the interaction of stellar winds and ionizing radiation of 12, 23, and 60 M⊙ stars within a cold neutral (CNM, n0 = 100 cm-3), warm neutral (WNM, n0 = 1, 10 cm-3) or warm ionized (WIM, n0 = 0.1 cm-3) medium. The FLASH simulations adopt the novel tree-based radiation transfer algorithm TREERAY. With the On-the-Spot approximation and a temperature-dependent recombination coefficient, it is coupled to a chemical network with radiative heating and cooling. In the homogeneous CNM, the total momentum injection ranges from 1.6× 104 to 4× 105 M⊙ km s-1 and is always dominated by the expansion of the ionized HII region. In the WIM, stellar winds dominate (2× 102 to 5× 103 M⊙ km s-1), while the input from radiation is small (˜ 102 M⊙ km s-1). The WNM (n0 = 1 cm-3) is a transition regime. Energetically, stellar winds couple more efficiently to the ISM (˜ 0.1 percent of wind luminosity) than radiation (< 0.001 percent of ionizing luminosity). For estimating the impact of massive stars, the strongly mass-dependent ratios of wind to ionizing luminosity and the properties of the ambient medium have to be considered.

  5. Experimental investigations of the effect the electrical conductivity of medium has on the performance of pH meters

    NASA Astrophysics Data System (ADS)

    Rodionov, A. K.; Karashchuk, S. A.

    2013-07-01

    Results from tests of pH meters carried out in ammonia media having low electric conductivity (less than 5.0 μS/cm) are presented. The check media for the tests were prepared in a special manner the use of which makes it possible to reproduce the pH value of solution with an error not exceeding ˜0.04pH in the range of electrical conductivities above 0.1 μS/cm. The instrument measurement error was determined at different electrical conductivities of medium. Different electrodes were tested, the majority of which were domestically produced ones intended for general industrial applications. Some results were also obtained for one dedicated electrode from a foreign manufacturer. The test results show that the instrument gives a biased pH value for such media. The bias has a random value, which nonetheless is stable in the majority of cases, depends on the electrical conductivity of medium being monitored, and may be quite essential for small electrical conductivities (0.5pH or more). A conclusion is drawn about the need to calibrate the instruments with respect to standard media having electrical conductivity close to that of the medium being monitored. Analytic relations characterizing the check media used fort tests (check solutions) are presented.

  6. [State of Fungal Lipases of Rhizopus microsporus, Penicillium sp. and Oospora lactis in Border Layers Water-Solid Phase and Factors Affecting Catalytic Properties of Enzymes].

    PubMed

    Khasanov, Kh T; Davranov, K; Rakhimov, M M

    2015-01-01

    We demonstrated that a change in the catalytic activity of fungal lipases synthesized by Rhizopus microsporus, Penicillium sp. and Oospora lactis and their ability to absorb on different sorbents depended on the nature of groups on the solid phase surface in the model systems water: lipid and water: solid phase. Thus, the stability of Penicillium sp. lipases increased 85% in the presence ofsorsilen or DEAE-cellulose, and 55% of their initial activity respectively was preserved. In the presence of silica gel and CM-cellulose, a decreased rate of lipid hydrolysis by Pseudomonas sp. enzymes was observed in water medium, and the hydrolysis rate increased by 2.4 and 1.5 times respectively in the presence of aminoaerosil and polykefamid. In an aqueous-alcohol medium, aminoaerosil and polykefamid decreased the rate of substrate hydrolysis by more than 30 times. The addition of aerosil to aqueous and aqueous-alcohol media resulted in an increase in the hydrolysis rate by 1.2-1.3 times. Sorsilen stabilized Penicillium sp. lipase activity at 40, 45, 50 and 55 degrees C. Either stabilization or inactivation of lipases was observed depending on the pH of the medium and the nature of chemical groups localized on the surface of solid phase. The synthetizing activity of lipases also changed depending on the conditions.

  7. Effect of the solution temperature in a singlet-oxygen generator on the formation of active medium in an ejector oxygen - iodine laser

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

    Zagidullin, M V; Nikolaev, V D; Svistun, M I

    2002-02-28

    The influence of the solution temperature in a singlet-oxygen generator on the formation of the active medium in the ejector oxygen - iodine laser is investigated. The following parameters of the active medium at the solution temperature -20{sup 0}C are obtained: the gain is 7.2 x 10{sup -3} cm{sup -1}, the Mach number is M=2, the temperature is 205 K, and the static pressure is 9.3 mmHg. As the solution temperature is increased to -4{sup 0}C, the gain decreases to 5 x 10{sup 3} cm{sup -1}, the Mach number decreases to 1.78, while the temperature and the static pressure increasemore » to 241 K and 10.7 mmHg, respectively. As the solution temperature increases from -20 to -4{sup 0}C, the losses in O{sub 2}({sup 1}{Delta}) increase by less than 20%, while the dissociation efficiency of molecular iodine decreases by less than 21%. (lasers, active media)« less

  8. Interaction of planetary nebulae with the interstellar medium

    NASA Technical Reports Server (NTRS)

    Borkowski, Kazimierz J.; Sarazin, Craig L.; Soker, Noam

    1990-01-01

    The interaction of a moving planetary nebula (PN) with the interstellar medium is considered. The PN shell is compressed first in the direction of the stellar motion. This produces a dipole asymmetry in the surface brightness of the nebula, typically at a nebular density of about 40/cu cm if the nebula is located in the Galactic plane. In the later stages of the interaction, this part of the shell is significantly decelerated with respect to the central star, and the PN becomes strongly asymmetric in shape. This distortion and the subsequent stripping of the nebular gas away from the central star typically occurs at a low nebular density of about 6/cu cm. The morphology of PNs with central stars whose proper motions exceed 0.015 arcsec/yr was examined, and it was found that many of the extended nebulae are interacting with the interstellar medium (ISM). The sample doubles the number of known PNs interacting with the ISM. The morphology of nearby PNs was examined, and a number of strongly asymmetric nebuale were found.

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

    Zagidullin, M V; Azyazov, V N; Malyshev, M S

    The kinetics of the processes occurring in an O{sub 2} – I{sub 2} – He – H{sub 2}O gas flow in which photodissociation of molecular iodine at a wavelength close to 500 nm and excitation of atomic iodine on the {sup 2}P{sub 1/2} – {sup 2}P{sub 3/2} transition by narrow-band radiation near 1315 nm are implemented successively has been analysed. It is shown that implementation of these processes allows one to form an oxygen – iodine medium with a high degree of dissociation of molecular iodine and a relative content of singlet oxygen O{sub 2}(a{sup 1}Δ) exceeding 10%. Having formedmore » a supersonic gas flow with a temperature ∼100 K from this medium, one can reach a small-signal gain of about 10{sup -2} cm{sup -1} on the {sup 2}P{sub 1/2} – {sup 2}P{sub 3/2} transition in iodine atoms. The specific power per unit flow cross section in the oxygen – iodine laser with this active medium may reach ∼100 W cm{sup -2}. (active media)« less

  10. Endometrial stem cells repair injured endometrium and induce angiogenesis via AKT and ERK pathways.

    PubMed

    Zhang, Yanling; Lin, Xiaona; Dai, Yongdong; Hu, Xiaoxiao; Zhu, Haiyan; Jiang, Yinshen; Zhang, Songying

    2016-11-01

    Intrauterine adhesions are common acquired endometrial syndromes secondary to endometrial injury, with limited effective therapies. Recently, several studies have reported that bone marrow stem cells (BMSCs) could repair injured endometrium in animal experiments. However, the role of stem cells in endometrial injury repair and its therapeutic mechanisms remain unclear. Here, we established mouse endometrial injury model and examined the benefit of human endometrial mesenchymal stem cells derived from menstrual blood (MenSCs) in restoration of injured endometrium. Injured endometrium exhibited significantly accelerated restoration at Day 7 after MenSCs transplantation, with increased endometrial thickness and microvessel density. Moreover, the fertility of mice with injured endometrium was improved, with higher conception rate (53.57% vs 14.29%, P = 0.014) and larger embryo number (3.1 ± 0.6 vs 0.9 ± 0.7, P = 0.030) in MenSCs group than control group, while no difference was found in undamaged horns between two groups. Conditioned medium from MenSCs (MenSCs-CM) could decrease H2O2-induced apoptosis of human umbilical vein endothelial cells (HUVECs) and promote proliferation, migration and angiogenesis. Angiogenesis effect of MenSCs-CM was also confirmed in Matrigel plug assay in mice. Furthermore, we discovered that MenSCs-CM could activate AKT and ERK pathways and induce the overexpression of eNOS, VEGFA, VEGFR1, VEGFR2 and TIE2 in HUVECs, which are critical in MenSCs-CM-induced angiogenesis. Angiogenesis induced by MenSCs-CM could be reversed by inhibitors of AKT and/or ERK. Taken together, we concluded that MenSCs could restore injured endometrium and improve the fertility of the endometrial injury mice, which was partially attributed to angiogenesis induced by MenSCs. © 2016 Society for Reproduction and Fertility.

  11. An in vitro method to test the safety and efficacy of low-level laser therapy (LLLT) in the healing of a canine skin model.

    PubMed

    Gagnon, Dominique; Gibson, Thomas W G; Singh, Ameet; zur Linden, Alex R; Kazienko, Jaimie E; LaMarre, Jonathan

    2016-04-08

    Low-level laser therapy (LLLT) has been used clinically as a treatment modality for a variety of medical conditions including wound-healing processes. It is an attractive and emerging method to enhance wound healing and improve clinical outcomes both in human and veterinary medicine. Despite the fact that the use of LLLT continues to gain in popularity, there is no universally accepted theory that defends all its cellular effects and beneficial biological processes in tissue repair. The present study was designed to evaluate the effect of LLLT on cellular migration and proliferation of cultured canine epidermal keratinocytes (CPEK) in an in vitro wound healing model. Keratinocyte migration and proliferation were assessed using a scratch migration assay and a proliferation assay, respectively. Fifteen independent replicates were performed for each assay. Canine epidermal keratinocyte cells exposed to LLLT with 0.1, 0.2, and 1.2 J/cm(2) migrated significantly more rapidly (p < 0.03) and showed significantly higher rates of proliferation (p < 0.0001) compared to non-irradiated cells cultured in the same medium and cells exposed to the higher energy dose of 10 J/cm(2). Irradiation with 10 J/cm(2) was characterized by decreased cellular migration and proliferation. These results revealed that LLLT has a measurable, dose-dependent effect on two different aspects of keratinocyte biology in vitro. In this in vitro wound-healing model, LLLT increased cellular migration and proliferation at doses of 0.1, 0.2, and 1.2 J/cm(2) while exposure to 10 J/cm(2) decreased cellular migration and proliferation. These data suggest that the beneficial effects of LLLT in vivo may be due, in part, to effects on keratinocyte behavior.

  12. Environmentally Acceptable Medium Caliber Ammunition Percussion Primers

    DTIC Science & Technology

    2007-10-31

    tetrazol-5-amino)-s-tetrazine or Bis-aminotetrazolyl-tetrazine CAD Cartridge Actuated Device CFR Code of Federal Regulations cm Centimeter cm3...approximately 120mg each, were placed on the Plexiglas slab . The piles were separated by a potassium chloride window estimated to transmit at least...for cartridge actuated device/propellant actuated device ( CAD /PAD) application. The NSWC-IH was working with the SDSMT and IMP in developing unique

  13. An Evaluation of Vegetated Roofing Technology: Application at Air Force Plant Four, Building 15

    DTIC Science & Technology

    2004-03-01

    layer of mineral wool , or recycled foam, or even installing a membrane that has water-absorbing crystals built-in. Adding more water retention...Sarnafil membrane has inherent root protection. Insulation: Eight cm thick hydroscopic mineral wool located under the waterproofing membrane...Drainage: Xero Drain, developed by Xeroflor. Growing medium: Four cm of mineral wool . This is a very lightweight material with excellent water

  14. Basic study on apoptosis induction into cancer cells U-937 and EL-4 by ultrasound exposure.

    PubMed

    Takeuchi, Shinichi; Udagawa, Yoshiko; Oku, Yumiko; Fujii, Takuma; Nishimura, Hiroyuki; Kawashima, Norimichi

    2006-12-22

    Recently, the low invasive cancer treatments with small aftereffects have been considered. We are studying on the suppression methods of cancer cell proliferation with ultrasound. Cancer cells of mouse T lymphoma (EL-4) have been used in our study. The human histitocytic lymphoma cells (U-937) was used in this time. The cancer cells were cultured in a culture medium of RPMI1640. The standing wave acoustic field was formed in a water tank of our ultrasound exposure system by a vibrating plate driven with a Langevine type transducer. The U-937 and EL-4 were exposed to ultrasound in the acoustic field with spatial average acoustic intensity of 350 mW/cm(2) at 150 kHz. The viable rate of EL-4 decreased with the lapse of culture time after ultrasound exposure. U-937 did not show the remarkable decrease tendency. The proliferation of U-937 which exposed to ultrasound with 700 mW/cm(2) was suppressed. It can be thought that apoptosis was induced in the cancer cells in this condition. We observed the morphological change on the U-937 exposed to ultrasound with this condition. The morphological changes by apoptosis like the shrink of cells, formation of apoptotic bodies etc. can be observed with an optical microscope and a phase contrast microscope.

  15. Sediment suspension and the dynamic mechanism during storms in the Yellow River Delta.

    PubMed

    Bian, Shuhua; Hu, Zjian; Liu, Jianqiang; Zhu, Zichen

    2016-12-01

    The suspension and hydrodynamic characteristics of the Yellow River Delta during storms were analyzed based on suspended samples obtained using automatic samplers during a storm event in the Yellow River Delta. Synchronous data for winds, waves, and tides were also collected from a nearby station. The results show that under wind speeds of 5-15 m/s and wave heights of 50-150 cm, the suspended content reached 5.7-49.6 kg/m 3 , which is 10-100 times higher than that under normal weather conditions. The medium diameter of suspended particles was 1.2-2.1 μm (8.9-9.7 Φ), which was approximately 1-2 Φ finer than that under normal weather conditions. During the early stages of the measurements, the sea level had risen by 50 cm owing to the storm, which was in addition to the tidal sea level change. We suggest that during the storms, the waves strengthened and the storm-induced sea level change, which was combined with tidal currents moving in the same direction, produced high-speed currents. This overcame the cohesive forces among the fine sediment particles and suspended a large amount of sediment. As a result, the suspended content increased markedly and the suspended particle size became finer. This explains the intense siltation and erosion of the Yellow River Delta during storms.

  16. Local-scale drivers of tree survival in a temperate forest.

    PubMed

    Wang, Xugao; Comita, Liza S; Hao, Zhanqing; Davies, Stuart J; Ye, Ji; Lin, Fei; Yuan, Zuoqiang

    2012-01-01

    Tree survival plays a central role in forest ecosystems. Although many factors such as tree size, abiotic and biotic neighborhoods have been proposed as being important in explaining patterns of tree survival, their contributions are still subject to debate. We used generalized linear mixed models to examine the relative importance of tree size, local abiotic conditions and the density and identity of neighbors on tree survival in an old-growth temperate forest in northeastern China at three levels (community, guild and species). Tree size and both abiotic and biotic neighborhood variables influenced tree survival under current forest conditions, but their relative importance varied dramatically within and among the community, guild and species levels. Of the variables tested, tree size was typically the most important predictor of tree survival, followed by biotic and then abiotic variables. The effect of tree size on survival varied from strongly positive for small trees (1-20 cm dbh) and medium trees (20-40 cm dbh), to slightly negative for large trees (>40 cm dbh). Among the biotic factors, we found strong evidence for negative density and frequency dependence in this temperate forest, as indicated by negative effects of both total basal area of neighbors and the frequency of conspecific neighbors. Among the abiotic factors tested, soil nutrients tended to be more important in affecting tree survival than topographic variables. Abiotic factors generally influenced survival for species with relatively high abundance, for individuals in smaller size classes and for shade-tolerant species. Our study demonstrates that the relative importance of variables driving patterns of tree survival differs greatly among size classes, species guilds and abundance classes in temperate forest, which can further understanding of forest dynamics and offer important insights into forest management.

  17. Oxygen Modulates Human Decidual Natural Killer Cell Surface Receptor Expression and Interactions with Trophoblasts1

    PubMed Central

    Wallace, Alison E.; Goulwara, Sonu S.; Whitley, Guy S.; Cartwright, Judith E.

    2014-01-01

    Decidual natural killer (dNK) cells have been shown to both promote and inhibit trophoblast behavior important for decidual remodeling in pregnancy and have a distinct phenotype compared to peripheral blood NK cells. We investigated whether different levels of oxygen tension, mimicking the physiological conditions of the decidua in early pregnancy, altered cell surface receptor expression and activity of dNK cells and their interactions with trophoblast. dNK cells were isolated from terminated first-trimester pregnancies and cultured in oxygen tensions of 3%, 10%, and 21% for 24 h. Cell surface receptor expression was examined by flow cytometry, and the effects of secreted factors in conditioned medium (CM) on the trophoblast cell line SGHPL-4 were assessed in vitro. SGHPL-4 cells treated with dNK cell CM incubated in oxygen tensions of 10% were significantly more invasive (P < 0.05) and formed endothelial-like networks to a greater extent (P < 0.05) than SGHPL-4 cells treated with dNK cell CM incubated in oxygen tensions of 3% or 21%. After 24 h, a lower percentage of dNK cells expressed CD56 at 21% oxygen (P < 0.05), and an increased percentage of dNK cells expressed NKG2D at 10% oxygen (P < 0.05) compared to other oxygen tensions, with large patient variation. This study demonstrates dNK cell phenotype and secreted factors are modulated by oxygen tension, which induces changes in trophoblast invasion and endovascular-like differentiation. Alterations in dNK cell surface receptor expression and secreted factors at different oxygen tensions may represent regulation of function within the decidua during the first trimester of pregnancy. PMID:25232021

  18. Local-Scale Drivers of Tree Survival in a Temperate Forest

    PubMed Central

    Wang, Xugao; Comita, Liza S.; Hao, Zhanqing; Davies, Stuart J.; Ye, Ji; Lin, Fei; Yuan, Zuoqiang

    2012-01-01

    Tree survival plays a central role in forest ecosystems. Although many factors such as tree size, abiotic and biotic neighborhoods have been proposed as being important in explaining patterns of tree survival, their contributions are still subject to debate. We used generalized linear mixed models to examine the relative importance of tree size, local abiotic conditions and the density and identity of neighbors on tree survival in an old-growth temperate forest in northeastern China at three levels (community, guild and species). Tree size and both abiotic and biotic neighborhood variables influenced tree survival under current forest conditions, but their relative importance varied dramatically within and among the community, guild and species levels. Of the variables tested, tree size was typically the most important predictor of tree survival, followed by biotic and then abiotic variables. The effect of tree size on survival varied from strongly positive for small trees (1–20 cm dbh) and medium trees (20–40 cm dbh), to slightly negative for large trees (>40 cm dbh). Among the biotic factors, we found strong evidence for negative density and frequency dependence in this temperate forest, as indicated by negative effects of both total basal area of neighbors and the frequency of conspecific neighbors. Among the abiotic factors tested, soil nutrients tended to be more important in affecting tree survival than topographic variables. Abiotic factors generally influenced survival for species with relatively high abundance, for individuals in smaller size classes and for shade-tolerant species. Our study demonstrates that the relative importance of variables driving patterns of tree survival differs greatly among size classes, species guilds and abundance classes in temperate forest, which can further understanding of forest dynamics and offer important insights into forest management. PMID:22347996

  19. Novel optical waveguides by in-depth controlled electronic damage with swift ions

    NASA Astrophysics Data System (ADS)

    Olivares, J.; García-Navarro, A.; Méndez, A.; Agulló-López, F.; García, G.; García-Cabañes, A.; Carrascosa, M.

    2007-04-01

    We review recent results on a novel method to modify crystalline dielectric materials and fabricate optical waveguides and integrated optics devices. It relies on irradiation with medium-mass high-energy ions (2-50 MeV) where the electronic stopping power is dominant over that one associated to nuclear collisions. By exploiting the processing capabilities of the method, novel optical structures can be achieved at moderate (1014 cm-2) and even low and ultralow (1012 cm-2) fluences. In particular, step-like waveguides with a high index jump Δn ∼ 0.1-0.2, guiding both ordinary and extraordinary modes, have been prepared with F and O ions (20 MeV) at moderate fluences. They present good non-linear and electrooptic perfomance and low losses. (1 dB/cm). Moreover, useful optical waveguiding has been also achieved at ultralow frequencies (isolated track regime), using Cl and Si ions (40-45 MeV). In this latter case, the individual amorphous nanotracks, whose radius increases with depth, create an effective optical medium causing optical trapping.

  20. Ps laser pulse induced stimulated Raman scattering of ammonium nitrate dissolved in water

    NASA Astrophysics Data System (ADS)

    Kumar, V. Rakesh; Kiran, P. Prem

    2018-04-01

    An intense picosecond laser pulse focused into a liquid medium generates a shock wave in the focal region. This shock wave while propagating into the medium varies the pressure and temperature of the liquid locally leading to the appearance of novel phases which are manifested by the appearance of Raman peaks. We present the phase changes of ammonium nitrate (AN) dissolved in water by studying the forward and backward stimulated Raman Scattering (FSRS and BSRS) signals due to propagation of 30 ps laser pulse induced shockwaves. The dominant peak corresponding to the NO3- symmetric stretching mode is observed with a Raman shift of 1045 cm-1 which represents phase IV of AN with an orthogonal crystalline structure. Apart from this peak, the dominant mode of liquid phase of water with a Raman shift of 3400 cm-1 and an ice VII peak at a Raman shift of 3050 cm-1 confirming the pressure of 10 GPa is observed. The effect of the concentration and input energy on the appearance of the phases will be presented.

  1. Thermal processing of bone: in vitro response of mesenchymal cells to bone-conditioned medium.

    PubMed

    Sawada, K; Caballé-Serrano, J; Schuldt Filho, G; Bosshardt, D D; Schaller, B; Buser, D; Gruber, R

    2015-08-01

    The autoclaving, pasteurization, and freezing of bone grafts to remove bacteria and viruses, and for preservation, respectively, is considered to alter biological properties during graft consolidation. Fresh bone grafts release paracrine-like signals that are considered to support tissue regeneration. However, the impact of the autoclaving, pasteurization, and freezing of bone grafts on paracrine signals remains unknown. Therefore, conditioned medium was prepared from porcine cortical bone chips that had undergone thermal processing. The biological properties of the bone-conditioned medium were assessed by examining the changes in expression of target genes in oral fibroblasts. The data showed that conditioned medium obtained from bone chips that had undergone pasteurization and freezing changed the expression of adrenomedullin, pentraxin 3, BTB/POZ domain-containing protein 11, interleukin 11, NADPH oxidase 4, and proteoglycan 4 by at least five-fold in oral fibroblasts. Bone-conditioned medium obtained from autoclaved bone chips, however, failed to change the expression of the respective genes. Also, when bone-conditioned medium was prepared from fresh bone chips, autoclaving blocked the capacity of bone-conditioned medium to modulate gene expression. These in vitro results suggest that pasteurization and freezing of bone grafts preserve the release of biologically active paracrine signals, but autoclaving does not. Copyright © 2015 International Association of Oral and Maxillofacial Surgeons. Published by Elsevier Ltd. All rights reserved.

  2. On the penetration of a hot diapir through a strongly temperature-dependent viscosity medium

    NASA Technical Reports Server (NTRS)

    Daly, S. F.; Raefsky, A.

    1985-01-01

    The ascent of a hot spherical body through a fluid with a strongly temperature-dependent viscosity has been studied using an axisymmetric finite element method. Numerical solutions range over Peclet numbers of 0.1 - 1000 from constant viscosity up to viscosity variations of 100,000. Both rigid and stress-free boundary conditions were applied at the surface of the sphere. The dependence of drag on viscosity variation was shown to have no dependence on the stress boundary condition except for a Stokes flow scaling factor. A Nusselt number parameterization based on the stress-free constant viscosity functional dependence on the Peclet number scaled by a parameter depending on the viscosity structure fits both stress-free and rigid boundary condition data above viscosity variations of 100. The temperature scale height was determined as a function of sphere radius. For the simple physical model studied in this paper pre-heating is required to reduce the ambient viscosity of the country rock to less than 10 to the 22nd sq cm/s in order for a 10 km diapir to penetrate a distance of several radii.

  3. Influence of climate variability on anchovy reproductive timing off northern Chile

    NASA Astrophysics Data System (ADS)

    Contreras-Reyes, Javier E.; Canales, T. Mariella; Rojas, Pablo M.

    2016-12-01

    We investigated the relationship between environmental variables and the Gonadosomatic Monthly Mean (GMM) index of anchovy (Engraulis ringens) to understand how the environment affects the dynamics of anchovy reproductive timing. The data examined corresponds to biological information collected from samples of the landings off northern Chile (18°21‧S, 24°00‧S) during the period 1990-2010. We used the Humboldt Current Index (HCI) and the Multivariate ENSO Index (MEI), which combine several physical-oceanographic factors in the Tropical and South Pacific regions. Using the GMM index, we studied the dynamics of anchovy reproductive timing at different intervals of length, specifically females with a length between 11.5 and 14 cm (medium class) and longer than 14 cm (large class). Seasonal Autoregressive Integrated Mobile Average (SARIMA) was used to predict missing observations. The trends of the environment and reproductive indexes were explored via the Breaks For Additive Season and Trend (BFAST) statistical technique and the relationship between these indexes via cross-correlation functions (CCF) analysis. Our results showed that the habitat of anchovy switched from cool to warm condition, which also influenced gonad development. This was revealed by two and three significant changes (breaks) in the trend of the HCI and MEI indexes, and two significant breaks in the GMM of each time series of anchovy females (medium and large). Negative cross-correlation between the MEI index and GMM of medium and large class females was found, indicating that as the environment gets warmer (positive value of MEI) a decrease in the reproductive activity of anchovy can be expected. Correlation between the MEI index and larger females was stronger than with medium females. Additionally, our results indicate that the GMM index of anchovy for both length classes reaches two maximums per year; the first from August to September and the second from December to January. The intensity (maximum GMM values at rise point) of reproductive activity was not equal though, with the August-September peak being the highest. We also discuss how the synchronicity between environment and reproductive timing, the negative correlation found between MEI and GMM indexes, and the two increases per year of anchovy GMM relate to previous studies. Based on these findings we propose ways to advance in the understanding of how anchovy synchronize gonad development with the environment.

  4. In vitro germination of zygotic embryos of hybrid BRS Manicoré (E. guineensis X E. oleifera).

    PubMed

    Bonetti, Keila A P; Quoirin, Marguerite; Quisen, Regina C; Lima, Suelen C S

    2016-01-01

    The interspecific oil palm hybrid BRS Manicoré (E. guineensis x E. oleifera) has superior agronomic characteristics. However, the germination rate is low (30%) and the process is slow when the seeds are sown in a conventional form. The purpose of this study was to optimize the in vitro germination of zygotic embryos of this hybrid comparing seed lots. The viability of zygotic embryos was evaluated by the tetrazolium test (0.075%) for 4 h. The embryos were cultured on MS and Y3 culture media, with and without the addition of NaH2PO4, as well as on MS, MS1/2 and N6 medium. In MS medium containing NaH2PO4, the germination rate was increased from 40 to 70% in comparison with the medium without sodium phosphate. The comparison between the culture media MS, MS 1/2, N6 and Y3 showed that 75% of zygotic embryos cultured in the Y3 medium formed whole plants (with roots and shoots defined), a higher percentage than embryos cultured on MS, MS 1/2 and N6 media (46, 35 and 17% respectively). In the same Y3 culture medium, the embryos were larger (36% ≥ 2 cm and 30% ≥ 5 cm) than in the other media. Results obtained by the tetrazolium test were similar to those of germination, showing the effect of the genotype of each seed lot. For the germination and development of plantlets it is essential to add NaH2PO4 to a culture medium containing no phosphate or with a low phosphate concentration.

  5. The Effects of ISM1 Medium on Embryo Quality and Outcomes of IVF/ICSI Cycles.

    PubMed

    Hassani, Fatemeh; Eftekhari-Yazdi, Poopak; Karimian, Leila; Rezazadeh Valojerdi, Mojtaba; Movaghar, Bahar; Fazel, Mohammad; Fouladi, Hamid Reza; Shabani, Fatemeh; Johansson, Lars

    2013-07-01

    The aim of this study is to investigate the effect of ISM1 culture medium on embryo development, quality and outcomes of in vitro fertilization/intracytoplasmic sperm injection (IVF/ICSI) cycles. This study compares culture medium commonly used in the laboratory setting for oocyte recovery and embryo development with a medium from MediCult. We have assessed the effects of these media on embryo development and newborn characteristics. In this prospective randomized study, fertilized oocytes from patients were randomly assigned to culture in ISM1 (MediCult, cycles: n=293) or routine lab culture medium (G-1TM v5; Vitrolife, cycles: n=290) according to the daily media schedule for oocyte retrieval. IVF or ICSI and embryo transfer were performed with either MediCult media or routine lab media. Embryo quality on days 2/3, cleavage, pregnancy and implantation rates, baby take home rate (BTHR), in addition to the weight and length of newborns were compared between groups. There were similar cleavage rates for ISM1 (86%) vs. G-1TM v5 (88%). We observed a significantly higher percentage of excellent embryos in ISM1 (42.7%) compared to G-1TM v5 (39%, p<0.05). Babies born after culture in ISM1 had both higher birth weight (3.03 kg) and length (48.8 cm) compared to G-1TM v5 babies that had a birth weight of 2.66 kg and a length of 46.0 cm (p<0.001 for both). This study suggests that ISM1 is a more effective culture medium in generating higher quality embryos, which may be reflected in the characteristics of babies at birth.

  6. The Effects of ISM1 Medium on Embryo Quality and Outcomes of IVF/ICSI Cycles

    PubMed Central

    Hassani, Fatemeh; Eftekhari-Yazdi, Poopak; Karimian, Leila; Rezazadeh Valojerdi, Mojtaba; Movaghar, Bahar; Fazel, Mohammad; Fouladi, Hamid Reza; Shabani, Fatemeh; Johansson, Lars

    2013-01-01

    Background: The aim of this study is to investigate the effect of ISM1 culture medium on embryo development, quality and outcomes of in vitro fertilization/intracytoplasmic sperm injection (IVF/ICSI) cycles. This study compares culture medium commonly used in the laboratory setting for oocyte recovery and embryo development with a medium from MediCult. We have assessed the effects of these media on embryo development and newborn characteristics. Materials and Methods: In this prospective randomized study, fertilized oocytes from patients were randomly assigned to culture in ISM1 (MediCult, cycles: n=293) or routine lab culture medium (G-1TM v5; Vitrolife, cycles: n=290) according to the daily media schedule for oocyte retrieval. IVF or ICSI and embryo transfer were performed with either MediCult media or routine lab media. Embryo quality on days 2/3, cleavage, pregnancy and implantation rates, baby take home rate (BTHR), in addition to the weight and length of newborns were compared between groups. Results: There were similar cleavage rates for ISM1 (86%) vs. G-1TM v5 (88%). We observed a significantly higher percentage of excellent embryos in ISM1 (42.7%) compared to G-1TM v5 (39%, p<0.05). Babies born after culture in ISM1 had both higher birth weight (3.03 kg) and length (48.8 cm) compared to G-1TM v5 babies that had a birth weight of 2.66 kg and a length of 46.0 cm (p<0.001 for both). Conclusion: This study suggests that ISM1 is a more effective culture medium in generating higher quality embryos, which may be reflected in the characteristics of babies at birth. PMID:24520472

  7. Identification of HgO/x/ species by matrix isolation spectroscopy

    NASA Technical Reports Server (NTRS)

    Butler, R.; Katz, S.; Snelson, A.; Stephens, J. B.

    1979-01-01

    The condensation of Hg atoms in an ozone-doped argon matrix gas with irradiation from a medium-pressure mercury arc lamp at 10 K resulted in the formation of several trapped HgO(x) species. Tentative identification of HgO has been made from absorption bands appearing in the visible and IR regions of the spectrum based on (O-16)2 and (O-18)2 isotope studies. The following spectroscopic constants were obtained: nu(00) = 14634 cm, nu(1) = 548 cm, and nu(11) = 676 cm. Some qualitative evidence was obtained for the existence of HgO2.

  8. Long-term culture of rat hippocampal neurons at low density in serum-free medium: combination of the sandwich culture technique with the three-dimensional nanofibrous hydrogel PuraMatrix.

    PubMed

    Kaneko, Ai; Sankai, Yoshiyuki

    2014-01-01

    The primary culture of neuronal cells plays an important role in neuroscience. There has long been a need for methods enabling the long-term culture of primary neurons at low density, in defined serum-free medium. However, the lower the cell density, the more difficult it is to maintain the cells in culture. Therefore, we aimed to develop a method for long-term culture of neurons at low density, in serum-free medium, without the need for a glial feeder layer. Here, we describe the work leading to our determination of a protocol for long-term (>2 months) primary culture of rat hippocampal neurons in serum-free medium at the low density of 3×10(4) cells/mL (8.9×10(3) cells/cm2) without a glial feeder layer. Neurons were cultured on a three-dimensional nanofibrous hydrogel, PuraMatrix, and sandwiched under a coverslip to reproduce the in vivo environment, including the three-dimensional extracellular matrix, low-oxygen conditions, and exposure to concentrated paracrine factors. We examined the effects of varying PuraMatrix concentrations, the timing and presence or absence of a coverslip, the timing of neuronal isolation from embryos, cell density at plating, medium components, and changing the medium or not on parameters such as developmental pattern, cell viability, neuronal ratio, and neurite length. Using our method of combining the sandwich culture technique with PuraMatrix in Neurobasal medium/B27/L-glutamine for primary neuron culture, we achieved longer neurites (≥3,000 µm), greater cell viability (≥30%) for 2 months, and uniform culture across the wells. We also achieved an average neuronal ratio of 97%, showing a nearly pure culture of neurons without astrocytes. Our method is considerably better than techniques for the primary culture of neurons, and eliminates the need for a glial feeder layer. It also exhibits continued support for axonal elongation and synaptic activity for long periods (>6 weeks).

  9. Bacterial lipoproteins and other factors released by Francisella tularensis modulate human neutrophil lifespan: Effects of a TLR1 SNP on apoptosis inhibition

    PubMed Central

    Kinkead, Lauren C.; Whitmore, Laura C.; McCracken, Jenna M.; Fletcher, Joshua R.; Ketelsen, Brandi B.; Kaufman, Justin W.; Jones, Bradley D.; Weiss, David S.; Barker, Jason H.

    2017-01-01

    Abstract Francisella tularensis infects several cell types including neutrophils, and aberrant neutrophil accumulation contributes to tissue destruction during tularaemia. We demonstrated previously that F. tularensis strains Schu S4 and live vaccine strain markedly delay human neutrophil apoptosis and thereby prolong cell lifespan, but the bacterial factors that mediate this aspect of virulence are undefined. Herein, we demonstrate that bacterial conditioned medium (CM) can delay apoptosis in the absence of direct infection. Biochemical analyses show that CM contained F. tularensis surface factors as well as outer membrane components. Our previous studies excluded roles for lipopolysaccharide and capsule in apoptosis inhibition, and current studies of [14C] acetate‐labelled bacteria argue against a role for other bacterial lipids in this process. At the same time, studies of isogenic mutants indicate that TolC and virulence factors whose expression requires FevR or MglA were also dispensable, demonstrating that apoptosis inhibition does not require Type I or Type VI secretion. Instead, we identified bacterial lipoproteins (BLPs) as active factors in CM. Additional studies of isolated BLPs demonstrated dose‐dependent neutrophil apoptosis inhibition via a TLR2‐dependent mechanism that is significantly influenced by a common polymorphism, rs5743618, in human TLR1. These data provide fundamental new insight into pathogen manipulation of neutrophil lifespan and BLP function. PMID:29063667

  10. Development of very compact soft X-ray lasers

    NASA Astrophysics Data System (ADS)

    Korobkin, Dmitriy Vladlenovich

    1999-10-01

    A powerful subpicosecond laser system, based on solid state (Ti:Al 2O3) front end and gas excimer (KrF*) amplifiers, has been developed. It is capable of producing 40-50 mJ pulses at 248 nm (285 fs pulse duration) with 2 Hz repetition rate. That radiation can be focused to intensities greater than 1017 W/cm2 and cause the optical field ionization of lithium. The system was used in experiments on gain generation in hydrogen-like Li III. Lasing action in hydrogen-like Li III 2-1 transition to ground state (13.5 nm) has been demonstrated for the first time. Gain G = 11 cm-1 in 5-mm long plasma column, created in LiF microcapillary, was measured. The observed gain-length product was GL = 5.5. In another set of experiments an inversion population between levels n = 3 and n = 2 in hydrogen-like BV was created using a low-energy compact laser system at 1 Hz repetition rate. Gain G = 17 cm-1 at 26.2 nm was measured in 3 mm long B2O3 microcapillary, which corresponds to GL = 5.1. The entire experimental setup can be fit on a single medium size optical table. Also propagation of laser radiation through plasma, created in microcapillaries has been investigated at various experimental conditions. Real-time plasma probing with low intensity HeNe laser beam has been performed.

  11. Mesenchymal Stem Cells Attenuate Cisplatin-Induced Nephrotoxicity in iNOS-Dependent Manner

    PubMed Central

    Simovic Markovic, Bojana; Gazdic, Marina; Arsenijevic, Aleksandar; Jovicic, Nemanja; Jeremic, Jovana; Djonov, Valentin; Arsenijevic, Nebojsa; Lukic, Miodrag L.

    2017-01-01

    Mesenchymal stem cells (MSCs) are, due to their immunomodulatory characteristics, utilized in therapy of immune-mediated diseases. We used murine model of cisplatin nephrotoxicity to explore the effects of MSCs on immune cells involved in the pathogenesis of this disease. Intraperitoneal application of MSCs significantly attenuated cisplatin nephrotoxicity, decreased inflammatory cytokines TNF-α and IL-17, and increased anti-inflammatory IL-10, IL-6, nitric oxide (NO), and kynurenine in sera of cisplatin-treated mice. MSC treatment significantly attenuated influx of leukocytes, macrophages, dendritic cells (DCs), neutrophils, CD4+ T helper (Th), and CD8+ cytotoxic T lymphocytes (CTLs) in damaged kidneys and attenuated the capacity of renal-infiltrated DCs, CD4+ Th, and CD8+ CTLs to produce TNF-α and IL-17. Similar effects were observed after intraperitoneal injection of MSC-conditioned medium (MSC-CM) indicating that MSCs exert their beneficial effects in paracrine manner. Inhibition of inducible nitric oxide synthase (iNOS) in MSC-CM resulted with increased number of TNF-α-producing DCs and IL-17-producing CTLs, decreased number of IL-10-producing tolerogenic DCs and regulatory CD4+FoxP3+ T cells, and completely diminished renoprotective effects of MSC-CM. In conclusion, MSCs, in iNOS-dependent manner, attenuated inflammation in cisplatin nephrotoxicity by reducing the influx and capacity of immune cells, particularly DCs and T lymphocytes, to produce inflammatory cytokines. PMID:28828008

  12. Surface Condensation of CO2 onto Kaolinite

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

    Schaef, Herbert T.; Glezakou, Vassiliki Alexandra; Owen, Antionette T.

    2014-02-11

    The fundamental adsorption behavior of gaseous and supercritical carbon dioxide (CO2) onto poorly crystalline kaolinite (KGa-2) at conditions relevant to geologic sequestration has been investigated using a quartz crystal microbalance (QCM) and density functional theory (DFT) methods. The QCM data indicated linear adsorption of CO2 (0-0.3 mmol CO2/g KGa-2) onto the kaolinite surface up through the gaseous state (0.186 g/cm3). However in the supercritical region, CO2 adsorption increases dramatically, reaching a peak (0.9-1.0 mmol CO2/g KGa-2) near 0.43 g/cm3, before declining rapidly to surface adsorption values equivalent or below gaseous CO2. This adsorption profile was not observed with He ormore » N2. Comparative density functional studies of CO2 interactions with kaolinite surface models rule out CO2 intercalation and confirm that surface adsorption is favored up to approximately 0.35 g/cm3 of CO2, showing distorted T-shaped CO2-CO2 clustering, typical of supercritical CO2 aggregation over the surface as the density increases. Beyond this point, the adsorption energy gain for any additional CO2 becomes less than the CO2 interaction energy (~0.2 eV) in the supercritical medium resulting in overall desorption of CO2 from the kaolinite surface.« less

  13. Inflammation increases NOTCH1 activity via MMP9 and is counteracted by Eicosapentaenoic Acid-free fatty acid in colon cancer cells

    PubMed Central

    Fazio, Chiara; Piazzi, Giulia; Vitaglione, Paola; Fogliano, Vincenzo; Munarini, Alessandra; Prossomariti, Anna; Milazzo, Maddalena; D’Angelo, Leonarda; Napolitano, Manuela; Chieco, Pasquale; Belluzzi, Andrea; Bazzoli, Franco; Ricciardiello, Luigi

    2016-01-01

    Aberrant NOTCH1 signalling is critically involved in multiple models of colorectal cancer (CRC) and a prominent role of NOTCH1 activity during inflammation has emerged. Epithelial to Mesenchymal Transition (EMT), a crucial event promoting malignant transformation, is regulated by inflammation and Metalloproteinase-9 (MMP9) plays an important role in this process. Eicosapentaenoic Acid (EPA), an omega-3 polyunsaturated fatty acid, was shown to prevent colonic tumors in different settings. We recently found that an extra-pure formulation of EPA as Free Fatty Acid (EPA-FFA) protects from colon cancer development in a mouse model of Colitis-Associated Cancer (CAC) through modulation of NOTCH1 signalling. In this study, we exposed colon cancer cells to an inflammatory stimulus represented by a cytokine-enriched Conditioned Medium (CM), obtained from THP1-differentiated macrophages. We found, for the first time, that CM strongly up-regulated NOTCH1 signalling and EMT markers, leading to increased invasiveness. Importantly, NOTCH1 signalling was dependent on MMP9 activity, upon CM exposure. We show that a non-cytotoxic pre-treatment with EPA-FFA antagonizes the effect of inflammation on NOTCH1 signalling, with reduction of MMP9 activity and invasiveness. In conclusion, our data suggest that, in CRC cells, inflammation induces NOTCH1 activity through MMP9 up-regulation and that this mechanism can be counteracted by EPA-FFA. PMID:26864323

  14. Tilted Magnetic Levitation Enables Measurement of the Complete Range of Densities of Materials with Low Magnetic Permeability.

    PubMed

    Nemiroski, Alex; Soh, Siowling; Kwok, Sen Wai; Yu, Hai-Dong; Whitesides, George M

    2016-02-03

    Magnetic levitation (MagLev) of diamagnetic or weakly paramagnetic materials suspended in a paramagnetic solution in a magnetic field gradient provides a simple method to measure the density of small samples of solids or liquids. One major limitation of this method, thus far, has been an inability to measure or manipulate materials outside of a narrow range of densities (0.8 g/cm(3) < ρ < 2.3 g/cm(3)) that are close in density to the suspending, aqueous medium. This paper explores a simple method-"tilted MagLev"-to increase the range of densities that can be levitated magnetically. Tilting the MagLev device relative to the gravitational vector enables the magnetic force to be decreased (relative to the magnetic force) along the axis of measurement. This approach enables many practical measurements over the entire range of densities observed in matter at ambient conditions-from air bubbles (ρ ≈ 0) to osmium and iridium (ρ ≈ 23 g/cm(3)). The ability to levitate, simultaneously, objects with a broad range of different densities provides an operationally simple method that may find application to forensic science (e.g., for identifying the composition of miscellaneous objects or powders), industrial manufacturing (e.g., for quality control of parts), or resource-limited settings (e.g., for identifying and separating small particles of metals and alloys).

  15. Identification of peptidases in highly pathogenic vs. weakly pathogenic Naegleria fowleri amebae.

    PubMed

    Vyas, Ishan K; Jamerson, Melissa; Cabral, Guy A; Marciano-Cabral, Francine

    2015-01-01

    Naegleria fowleri, a free-living ameba, is the causative agent of Primary Amebic Meningoencephalitis. Highly pathogenic mouse-passaged amebae (Mp) and weakly pathogenic axenically grown (Ax) N. fowleri were examined for peptidase activity. Zymography and azocasein peptidase activity assays demonstrated that Mp and Ax N. fowleri exhibited a similar peptidase pattern. Prominent for whole cell lysates, membranes and conditioned medium (CM) from Mp and Ax amebae was the presence of an activity band of approximately 58 kDa that was sensitive to E64, a cysteine peptidase inhibitor. However, axenically grown N. fowleri demonstrated a high level of this peptidase activity in membrane preparations. The inhibitor E64 also reduced peptidase activity in ameba-CM consistent with the presence of secreted cysteine peptidases. Exposure of Mp amebae to E64 reduced their migration through matrigel that was used as an extracellular matrix, suggesting a role for cysteine peptidases in invasion of the central nervous system (CNS). The collective results suggest that the profile of peptidases is not a discriminative marker for distinguishing Mp from Ax N. fowleri. However, the presence of a prominent level of activity for cysteine peptidases in N. fowleri membranes and CM, suggests that these enzymes may serve to facilitate passage of the amebae into the CNS. © 2014 The Author(s) Journal of Eukaryotic Microbiology © 2014 International Society of Protistologists.

  16. Inflammation increases NOTCH1 activity via MMP9 and is counteracted by Eicosapentaenoic Acid-free fatty acid in colon cancer cells.

    PubMed

    Fazio, Chiara; Piazzi, Giulia; Vitaglione, Paola; Fogliano, Vincenzo; Munarini, Alessandra; Prossomariti, Anna; Milazzo, Maddalena; D'Angelo, Leonarda; Napolitano, Manuela; Chieco, Pasquale; Belluzzi, Andrea; Bazzoli, Franco; Ricciardiello, Luigi

    2016-02-11

    Aberrant NOTCH1 signalling is critically involved in multiple models of colorectal cancer (CRC) and a prominent role of NOTCH1 activity during inflammation has emerged. Epithelial to Mesenchymal Transition (EMT), a crucial event promoting malignant transformation, is regulated by inflammation and Metalloproteinase-9 (MMP9) plays an important role in this process. Eicosapentaenoic Acid (EPA), an omega-3 polyunsaturated fatty acid, was shown to prevent colonic tumors in different settings. We recently found that an extra-pure formulation of EPA as Free Fatty Acid (EPA-FFA) protects from colon cancer development in a mouse model of Colitis-Associated Cancer (CAC) through modulation of NOTCH1 signalling. In this study, we exposed colon cancer cells to an inflammatory stimulus represented by a cytokine-enriched Conditioned Medium (CM), obtained from THP1-differentiated macrophages. We found, for the first time, that CM strongly up-regulated NOTCH1 signalling and EMT markers, leading to increased invasiveness. Importantly, NOTCH1 signalling was dependent on MMP9 activity, upon CM exposure. We show that a non-cytotoxic pre-treatment with EPA-FFA antagonizes the effect of inflammation on NOTCH1 signalling, with reduction of MMP9 activity and invasiveness. In conclusion, our data suggest that, in CRC cells, inflammation induces NOTCH1 activity through MMP9 up-regulation and that this mechanism can be counteracted by EPA-FFA.

  17. Image analysis method for the measurement of water saturation in a two-dimensional experimental flow tank

    NASA Astrophysics Data System (ADS)

    Belfort, Benjamin; Weill, Sylvain; Lehmann, François

    2017-04-01

    A novel, non-invasive imaging technique that determines 2D maps of water content in unsaturated porous media is presented. This method directly relates digitally measured intensities to the water content of the porous medium. This method requires the classical image analysis steps, i.e., normalization, filtering, background subtraction, scaling and calibration. The main advantages of this approach are that no calibration experiment is needed and that no tracer or dye is injected into the flow tank. The procedure enables effective processing of a large number of photographs and thus produces 2D water content maps at high temporal resolution. A drainage / imbibition experiment in a 2D flow tank with inner dimensions of 40 cm x 14 cm x 6 cm (L x W x D) is carried out to validate the methodology. The accuracy of the proposed approach is assessed using numerical simulations with a state-of-the-art computational code that solves the Richards. Comparison of the cumulative mass leaving and entering the flow tank and water content maps produced by the photographic measurement technique and the numerical simulations demonstrate the efficiency and high accuracy of the proposed method for investigating vadose zone flow processes. Application examples to a larger flow tank with various boundary conditions are finally presented to illustrate the potential of the methodology.

  18. SU-F-T-369: Validation of Monte-Carlo Beam Model for a Range of Small Fields in Heterogeneous Medium - A Measurement Based Study

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

    Karthikeyan, N; Bharathiya University, Coimbatore, Tamilnadu; Ganesh, KM

    Purpose: To validate the Monaco montecorlo beam model for a range of small field in the heterogeneous medium. Methods: A in-house phantom with three different medium of Foam, PMMA and derlin resembling the densities of lung, soft tissue, and bone was used for the study. The field sizes of 8, 16, 24, 32 and 48mm were studied for the validation of montecarlo algorithm using 0.01cc volume ionchamber and gafchromic films. The 6MV photon beam from Elekta Beam modulator was used with 100cm SAD setup. The outputs were measured at the depth of 5, 10 and 20mm in every second mediummore » with 3cm buildup of first medium for the interface of lung-bone, lung-soft tissue, soft tissue-bone, bone-lung and soft tissue-lung. Similarly, the 2D dose analysis with gamma criteria of 2%2mm were done at the same depths using gafchromic film. For all the measurements 10.4×10.4cm were taken as reference to which the other field sizes were compared. Monaco TPSv.3.20 was used to calculate the dose distribution for all the simulated measurement setups. Results: The average maximum difference among the field sizes of 8, 16, 24, 32 and 48mm at the depth of 5mm in second medium with the interface of lung-bone, lung-soft tissue, soft tissue-bone, bone-lung and soft tissue-lung were observed as 1.29±0.14%, 0.49±0.16%, 0.87±0.23%, 0.92±0.11%, 1.01±0.19% respectively. The minimum and maximum variation of dose among different materials for the smallest field size of 8mm were observed as 0.23% and 1.67% respectively. The 2D analysis showed the average gamma passing of 98.9±0.5%. The calculated two-tailed P-value were showed insignificance with values of 0.562 and 0.452 for both ionchamber and film measurements. Conclusion: The accuracy of dose calculation for the small fields in Monaco Montecarlo TPS algorithm was validated in different inhomogeneous medium and found the results were well correlated with measurement data.« less

  19. Mycelial mass production of fungi Duddingtonia flagrans and Monacrosporium thaumasium under different culture conditions

    PubMed Central

    2013-01-01

    Background Duddingtonia flagrans and Monacrosporium thaumasium are promising fungus species in veterinary biological control of gastrointestinal nematodes because of their production capacity of fungal structures (conidia and/or chlamydospores), growth efficiency in laboratory solid media and especially their predatory capacity. However, their large-scale production remains a challenge. This work aimed at evaluating the mycelial mass production of D. flagrans (AC001 and CG722) and M. thaumasium (NF34A) nematophagous fungi under different culture conditions. Results The results did not present significant differences (p > 0.05) in mycelia mass production between the isolates cultured under pH 4.0. Furthermore, after 168 hrs., the isolate CG722 presented a lower production of mycelial mass in medium CM (corn meal) (p < 0.05). Conclusion We therefore concluded the use of culture media SD (soy dextrose) and CG (corn grits) at pH values between 6.0 and 7.0 is suitable for high mycelial mass production of D. flagrans and M. thaumasium. PMID:23985336

  20. Benzene Formation on Interstellar Icy Mantles Containing Propargyl Alcohol

    NASA Astrophysics Data System (ADS)

    Sivaraman, B.; Mukherjee, R.; Subramanian, K. P.; Banerjee, S. B.

    2015-01-01

    Propargyl alcohol (CHCCH2OH) is a known stable isomer of the propenal (CH2CHCHO) molecule that was reported to be present in the interstellar medium (ISM). At astrochemical conditions in the laboratory, icy layers of propargyl alcohol grown at 85 K were irradiated by 2 keV electrons and probed by a Fourier Transform InfraRed spectrometer in the mid-infrared (IR) region, 4000-500 cm-1. Propargyl alcohol ice under astrochemical conditions was studied for the first time; therefore, IR spectra of reported amorphous (85 K) and crystalline (180 K) propargyl alcohol ices can be used to detect its presence in the ISM. Moreover, our experiments clearly show benzene (C6H6) formation to be the major product from propargyl alcohol irradiation, confirming the role of propargyl radicals (C3H3) formed from propargyl alcohol dissociation that was long expected based on theoretical modeling to effectively synthesize C6H6 in the interstellar icy mantles.

  1. Copper oxide assisted cysteine hierarchical structures for immunosensor application

    NASA Astrophysics Data System (ADS)

    Pandey, Chandra Mouli; Sumana, Gajjala; Tiwari, Ida

    2014-09-01

    The present work describes the promising electrochemical immunosensing strategy based on copper (II) assisted hierarchical cysteine structures (CuCys) varying from star to flower like morphology. The CuCys having average size of 10 μm have been synthesised using L-Cysteine as initial precursor in presence of copper oxide under environmentally friendly conditions in aqueous medium. To delineate the synthesis mechanism, detailed structural investigations have been carried out using characterization techniques such as X-ray diffraction, transmission electron microscopy, and Fourier transform infrared spectroscopy. The electrochemical behaviour of self-assembled CuCys on gold electrode shows surface controlled electrode reaction with an apparent electron transfer rate constant of 3.38 × 10-4 cm s-1. This innovative platform has been utilized to fabricate an immunosensor by covalently immobilizing monoclonal antibodies specific for Escherichia coli O157:H7 (E. coli). Under the optimal conditions, the fabricated immunosensor is found to be sensitive and specific for the detection of E. coli with a detection limit of 10 cfu/ml.

  2. Hypoxic-Preconditioned Bone Marrow Stem Cell Medium Significantly Improves Outcome After Retinal Ischemia in Rats

    PubMed Central

    Roth, Steven; Dreixler, John C.; Mathew, Biji; Balyasnikova, Irina; Mann, Jacob R.; Boddapati, Venkat; Xue, Lai; Lesniak, Maciej S.

    2016-01-01

    Purpose We have previously demonstrated the protective effect of bone marrow stem cell (BMSC)-conditioned medium in retinal ischemic injury. We hypothesized here that hypoxic preconditioning of stem cells significantly enhances the neuroprotective effect of the conditioned medium and thereby augments the protective effect in ischemic retina. Methods Rats were subjected to retinal ischemia by increasing intraocular pressure to 130 to 135 mm Hg for 55 minutes. Hypoxic-preconditioned, hypoxic unconditioned, or normoxic medium was injected into the vitreous 24 hours after ischemia ended. Recovery was assessed 7 days after injections by comparing electroretinography measurements, histologic examination, and apoptosis (TUNEL, terminal deoxynucleotidyl transferase–mediated dUTP nick end labeling assay). To compare proteins secreted into the medium in the groups and the effect of hypoxic exposure, we used rat cytokine arrays. Results Eyes injected with hypoxic BMSC–conditioned medium 24 hours after ischemia demonstrated significantly enhanced return of retinal function, decreased retinal ganglion cell layer loss, and attenuated apoptosis compared to those administered normoxic or hypoxic unconditioned medium. Hypoxic-preconditioned medium had 21 significantly increased protein levels compared to normoxic medium. Conclusions The medium from hypoxic-preconditioned BMSCs robustly restored retinal function and prevented cell loss after ischemia when injected 24 hours after ischemia. The protective effect was even more pronounced than in our previous studies of normoxic conditioned medium. Prosurvival signals triggered by the secretome may play a role in this neuroprotective effect. PMID:27367588

  3. Measured neutron and gamma spectra from californium-252 in a tissue-equivalent medium.

    PubMed

    Elson, H R; Stupar, T A; Shapiro, A; Kereiakes, J G

    1979-01-01

    A method of experimentally obtaining both neutron and gamma-ray spectra in a scattering medium is described. The method utilizes a liquid-organic scintillator (NE-213) coupled with a pulse-shape discrimination circuit. This allows the separation of the neutron-induced pulse-height data from the gamma-ray pulse-height data. Using mathematical unfolding techniques, the two sets of pulse-height data were transformed to obtain the neutron and gamma-ray energy spectra. A small spherical detector was designed and constructed to reduce the errors incurred by attempting spectral measurements in a scattering medium. Demonstration of the utility of the system to obtain the neutron and gamma-ray spectra in a scattering medium was performed by characterizing the neutron and gamma-ray spectra at various sites about a 3.7-microgram (1.5 cm active length) californium-252 source in a tissue-equivalent medium.

  4. Transplantation of Reprogrammed Autologous Stem Cells for Chronic Pain and Drug Abuse

    DTIC Science & Technology

    2013-10-01

    endogenous analgesic substances, including enkephalins, catecholamines, gamma aminobutyric acid , indolalkylamines, and other neuropeptides (7, 8). The...containing Dulbecco’s modified eagle medium /F12 (DMEM/F12, 1:1; Gibco, Grand Island, NY) supplemented with 10% (v/v) fetal bovine serum (FBS, Sigma...Cambridge, MA) at a concentration of 1x10 5 cells/cm 2 and cultured in 20 ml growth medium consisting of DMEM (Gibco, Grand Island, NY), an antibiotic

  5. Osteopontin-c mediates the upregulation of androgen responsive genes in LNCaP cells through PI3K/Akt and androgen receptor signaling.

    PubMed

    Tilli, Tatiana Martins; Ferreira, Luciana Bueno; Gimba, Etel Rodrigues Pereira

    2015-04-01

    Androgen receptor (AR) signaling is a key pathway modulating prostate cancer (PCa) progression. Several steps in this pathway have been investigated in order to propose novel treatment strategies for advanced PCa. Total osteopontin (OPN) has been described as a biomarker for PCa, in addition to its role in activating the progression of this tumor. Based on the known effects of the OPNc splice variant on PCa progression, the present study investigated whether this isoform can also modulate AR signaling. In order to test this, an in vitro model was used in which LNCaP cells were cultured in the presence of conditioned medium (CM) secreted by PCa cells overexpressing OPNc (OPNc-CM). The activation of AR signaling was evaluated by measuring the expression levels of AR-responsive genes (ARGs) using quantitative polymerase chain reaction and specific oligonucleotides. The data demonstrated that all nine tested ARGs ( Fgf8 , TMPRSS2 , Greb1 , Cdk2 , Ndrg1 , Cdk1 , Pmepa1 , Psa and Ar ) are significantly upregulated in response to OPNc-CM compared with LNCaP cells cultured in CM secreted by control cells transfected with empty expression vector. The specific involvement of OPNc was demonstrated by depleting OPNc from OPNc-CM using an anti-OPNc neutralizing antibody. In addition, by using a phosphoinositide 3-kinase (PI3K)-specific inhibitor and AR antagonists, such as flutamide and bicalutamide, it was also observed that upregulation of ARGs in response to OPNc-CM involves PI3K signaling and depends on the AR. In conclusion, these data indicated that OPNc is able to activate AR signaling through the PI3K pathway and the AR. These data further corroborate our previous data, revealing the OPNc splice variant to be a key molecule that is able to modulate key signaling pathways involved in PCa progression.

  6. Osteopontin-c mediates the upregulation of androgen responsive genes in LNCaP cells through PI3K/Akt and androgen receptor signaling

    PubMed Central

    TILLI, TATIANA MARTINS; FERREIRA, LUCIANA BUENO; GIMBA, ETEL RODRIGUES PEREIRA

    2015-01-01

    Androgen receptor (AR) signaling is a key pathway modulating prostate cancer (PCa) progression. Several steps in this pathway have been investigated in order to propose novel treatment strategies for advanced PCa. Total osteopontin (OPN) has been described as a biomarker for PCa, in addition to its role in activating the progression of this tumor. Based on the known effects of the OPNc splice variant on PCa progression, the present study investigated whether this isoform can also modulate AR signaling. In order to test this, an in vitro model was used in which LNCaP cells were cultured in the presence of conditioned medium (CM) secreted by PCa cells overexpressing OPNc (OPNc-CM). The activation of AR signaling was evaluated by measuring the expression levels of AR-responsive genes (ARGs) using quantitative polymerase chain reaction and specific oligonucleotides. The data demonstrated that all nine tested ARGs (Fgf8, TMPRSS2, Greb1, Cdk2, Ndrg1, Cdk1, Pmepa1, Psa and Ar) are significantly upregulated in response to OPNc-CM compared with LNCaP cells cultured in CM secreted by control cells transfected with empty expression vector. The specific involvement of OPNc was demonstrated by depleting OPNc from OPNc-CM using an anti-OPNc neutralizing antibody. In addition, by using a phosphoinositide 3-kinase (PI3K)-specific inhibitor and AR antagonists, such as flutamide and bicalutamide, it was also observed that upregulation of ARGs in response to OPNc-CM involves PI3K signaling and depends on the AR. In conclusion, these data indicated that OPNc is able to activate AR signaling through the PI3K pathway and the AR. These data further corroborate our previous data, revealing the OPNc splice variant to be a key molecule that is able to modulate key signaling pathways involved in PCa progression. PMID:25789054

  7. Accelerated differentiation of human induced pluripotent stem cells to blood-brain barrier endothelial cells.

    PubMed

    Hollmann, Emma K; Bailey, Amanda K; Potharazu, Archit V; Neely, M Diana; Bowman, Aaron B; Lippmann, Ethan S

    2017-04-13

    Due to their ability to limitlessly proliferate and specialize into almost any cell type, human induced pluripotent stem cells (iPSCs) offer an unprecedented opportunity to generate human brain microvascular endothelial cells (BMECs), which compose the blood-brain barrier (BBB), for research purposes. Unfortunately, the time, expense, and expertise required to differentiate iPSCs to purified BMECs precludes their widespread use. Here, we report the use of a defined medium that accelerates the differentiation of iPSCs to BMECs while achieving comparable performance to BMECs produced by established methods. Induced pluripotent stem cells were seeded at defined densities and differentiated to BMECs using defined medium termed E6. Resultant purified BMEC phenotypes were assessed through trans-endothelial electrical resistance (TEER), fluorescein permeability, and P-glycoprotein and MRP family efflux transporter activity. Expression of endothelial markers and their signature tight junction proteins were confirmed using immunocytochemistry. The influence of co-culture with astrocytes and pericytes on purified BMECs was assessed via TEER measurements. The robustness of the differentiation method was confirmed across independent iPSC lines. The use of E6 medium, coupled with updated culture methods, reduced the differentiation time of iPSCs to BMECs from thirteen to 8 days. E6-derived BMECs expressed GLUT-1, claudin-5, occludin, PECAM-1, and VE-cadherin and consistently achieved TEER values exceeding 2500 Ω × cm 2 across multiple iPSC lines, with a maximum TEER value of 4678 ± 49 Ω × cm 2 and fluorescein permeability below 1.95 × 10 -7 cm/s. E6-derived BMECs maintained TEER above 1000 Ω × cm 2 for a minimum of 8 days and showed no statistical difference in efflux transporter activity compared to BMECs differentiated by conventional means. The method was also found to support long-term stability of BMECs harboring biallelic PARK2 mutations associated with Parkinson's Disease. Finally, BMECs differentiated using E6 medium responded to inductive cues from astrocytes and pericytes and achieved a maximum TEER value of 6635 ± 315 Ω × cm 2 , which to our knowledge is the highest reported in vitro TEER value to date. Given the accelerated differentiation, equivalent performance, and reduced cost to produce BMECs, our updated methods should make iPSC-derived in vitro BBB models more accessible for a wide variety of applications.

  8. THE FRACTIONAL IONIZATION OF THE WARM NEUTRAL INTERSTELLAR MEDIUM

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

    Jenkins, Edward B., E-mail: ebj@astro.princeton.edu

    2013-02-10

    When the neutral interstellar medium is exposed to extreme-ultraviolet and soft X-ray radiation, the argon atoms in it are far more susceptible to being ionized than the hydrogen atoms. We make use of this fact to determine the level of ionization in the nearby warm neutral medium. By analyzing Far-Ultraviolet Spectroscopic Explorer observations of ultraviolet spectra of 44 hot subdwarf stars a few hundred parsecs away from the Sun, we can compare column densities of Ar I to those of O I, where the relative ionization of oxygen can be used as a proxy for that of hydrogen. The measuredmore » deficiency [Ar I/O I]=-0.427{+-}0.11 dex below the expectation for a fully neutral medium implies that the electron density n(e) Almost-Equal-To 0.04 cm{sup -3} if n(H) = 0.5 cm{sup -3}. This amount of ionization is considerably larger than what we expect from primary photoionizations resulting from cosmic rays, the diffuse X-ray background, and X-ray emitting sources within the medium, along with the additional ionizations caused by energetic secondary photoelectrons, Auger electrons, and photons from helium recombinations. We favor an explanation that bursts of radiation created by previous, nearby supernova remnants that have faded by now may have elevated the ionization, and the gas has not yet recombined to a quiescent level. A different alternative is that the low-energy portion of the soft X-ray background is poorly shielded by the H I because it is frothy and has internal pockets of very hot, X-ray emitting gases.« less

  9. Effect of low-level diode laser on proliferation and osteogenic differentiation of dental pulp stem cells

    NASA Astrophysics Data System (ADS)

    Tabatabaei, Fahimeh S.; Torshabi, Maryam; Mojahedi Nasab, Masoud; Khosraviani, Keikhosro; Khojasteh, Arash

    2015-09-01

    This study assessed the effect of low-level laser irradiation (LLLI) on the proliferation and osteogenic differentiation of dental pulp stem cells (DPSCs). DPSCs were exposed to 810 nm laser light (0.1, 0.2, or 0.3 J cm-2) for 7 d (60 s daily). The negative control group (cells in regular medium) and positive control group (cells in osteogenic medium (OM)) were not lased. One group of cells in OM was irradiated with laser operated at 0.2 J cm-2. Cell viability was evaluated at 24 h and one week after the last day of laser irradiation using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Osteogenic differentiation was assessed using real-time reverse transcriptase polymerase chain reaction (RT-PCR) and alizarin Red S staining. Cell proliferation was not affected by laser irradiation at 24 h except in one group (cells in OM exposed to laser at 0.2 J cm-2). However, one week after the last day of laser irradiation, it was significantly increased in groups exposed to laser at 0.1 or 0.2 J cm-2 and decreased in groups containing OM (P  <  0.05). Osteoblast marker expression was observed in groups containing OM. LLLI at 0.2 J cm-2 dramatically enhanced cell differentiation. Laser at 0.3 J cm-2 increased bone sialoprotein (BSP) and decreased alkaline phosphatase (ALP). Mineralized nodules were only observed in groups containing OM. Considering these findings, LLLI may be used as a novel approach for preconditioning of DPSCs in vitro prior to bone tissue engineering.

  10. Preculturing effect of thidiazuron on in vitro shoot multiplication and micropropagation round in Capparis decidua (Forsk.) an important multipurpose plant.

    PubMed

    Bukhari, Najat A W; Siddique, Iram; Perveen, Kahkashan

    2016-09-01

    An efficient protocol was developed for clonal multiplication of an important shrub: Capparis decidua (Forsk.) Edgew, through in vitro shoot induction and multiplication from nodal explants. Pretreatment of nodal explants in a liquid Murashige and Skoog (MS) medium augmented with various thidiazuron (TDZ) concentrations at relatively high levels (5-100 μM) for different time duration (4, 8, 12 and 16 d), proved a significant approach for in vitro shoot production. After an initial exposure time to TDZ, nodal explants were inoculated onto a MS basal medium devoid of TDZ for further induction and proliferation. The highest regeneration rate (85%), average number of shoots/explant (8.7 ± 0.22) and maximum shoot length (3.9 ± 0.33 cm) were obtained from the nodal explants exposed to 50 μM TDZ for 8 d. The nodal explants excised from the proliferated cultures of TDZ (50 μM) for 8 d were used as explants and showed an enhancement rate after next three round of in vitro propagation. Best results for rooting was obtained by ex vitro treatment of shoots with 200 μM indole-3-butyric acid (IBA) for 20 min. as it produced an average of 5.7 ± 0.41 roots per microshoot with 4.4 ± 0.39 cm root length in 84% shoots. Different planting substrates was tested for maximum survival of hardening off micropropagated plantlets and soilrite proved most effective than others as 97.1 ± 7.21 plantlets survived. All micropropagated plants grew well in natural conditions and showed similar morphology to the mother plant.

  11. Antibacterial Effect of Human Mesenchymal Stem Cells Is Mediated in Part from Secretion of the Antimicrobial Peptide LL-37

    PubMed Central

    Krasnodembskaya, Anna; Song, Yuanlin; Fang, Xiaohui; Gupta, Naveen; Serikov, Vladimir; Lee, Jae-Woo; Matthay, Michael A.

    2012-01-01

    Recent in vivo studies indicate that mesenchymal stem cells (MSCs) may have beneficial effects in the treatment of sepsis induced by bacterial infection. Administration of MSCs in these studies improved survival and enhanced bacterial clearance. The primary objective of this study was to test the hypothesis that human MSCs possessed intrinsic antimicrobial properties. We studied the effect of human MSCs derived from bone marrow on the bacterial growth of Gram-negative (Escherichia coli and Pseudomonas aeruginosa) and Gram-positive (Staphylococcus aureus) bacteria. MSCs as well as their conditioned medium (CM) demonstrated marked inhibition of bacterial growth in comparison with control medium or normal human lung fibroblasts (NHLF). Analysis of expression of major antimicrobial peptides indicated that one of the factors responsible for the antimicrobial activity of MSC CM against Gram-negative bacteria was the human cathelicidin antimicrobial peptide, hCAP-18/LL-37. Both m-RNA and protein expression data showed that the expression of LL-37 in MSCs increased after bacterial challenge. Using an in vivo mouse model of E. coli pneumonia, intratracheal administration of MSCs reduced bacterial growth (in colony-forming unit) in the lung homogenates and in the bronchoalveolar lavage (BAL) fluid, and administration of MSCs simultaneously with a neutralizing antibody to LL-37 resulted in a decrease in bacterial clearance. In addition, the BAL itself from MSC-treated mice had a greater antimicrobial activity in comparison with the BAL of phosphate buffered saline (PBS)-treated mice. Human bone marrow-derived MSCs possess direct antimicrobial activity, which is mediated in part by the secretion of human cathelicidin hCAP-18/LL-37. PMID:20945332

  12. Chromosomal studies on 2 mL of celomic fluid obtained during the fifth week of development in the timed-pregnant baboon model.

    PubMed

    Santolaya-Forgas, Joaquin; De Leon-Luis, Juan; Shen, Zhion; McCorquodale, Maureen

    2005-09-01

    To determine if chromosomal studies could be performed using 2 mL of celomicfluid obtained during the fifth postfertilization week in pregnant baboons. Nine ultrasound-guided celocenteses were performed. The initial 0.5 mL of celomic fluid was discarded to decrease maternal cell contamination. Approximately 2 mL of celomic fluid was then collected. The fluid was centrifuged and the supernatant removed to a final volume of 0.5 mL. The celomic fluid sample was placed in either a small plastic flaskette chamber slide with a mix of 0.5 mL celomic fluid, 1 mL of Amniomax, and 1 mL of usedfibroblast culture medium to spread on the entire surface (n=4), or a 3.5 x 1-cm plastic Petri dish with a 24 x 30-mm glass coverslip to keep the 0.5 mL celomic fluid mixed with 1 mL of Amniomax (Invitrogen, Carlsbad, California) within a 1 cm2 area (n=5). The medium was changed on day 5 and thereafter every second to third day. The cells were harvested when the number of cells appeared sufficient for chromosomal analysis. Standard chromosomal studies were possible in 5 of the 9 celomicfluid samples. Mean (+/-SD) celomic fluid volume used for culture was 1.85 +/- 0.3 mL. Mean (+/-SD) time to karyotype result was 18.8 +/- 1.8 days. The findings of this study suggest that there are living cells at 36-42 days of embryonic development in the extraembryonic celomic fluid of primates and that they can be cultured for chromosomal studies. However, significant improvements in understanding the biology of cells present at 5 weeks after fertilization in celomic fluid are needed to improve culture conditions.

  13. Automotive airborne brake wear debris nanoparticles and cytokinesis-block micronucleus assay in peripheral blood lymphocytes: A pilot study

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

    Kazimirova, Alena, E-mail: alena.kazimirova@szu.sk; Peikertova, Pavlina; IT4Innovations, VŠB – Technical University of Ostrava, 17. listopadu 15, 708 33 Ostrava-Poruba

    Motor vehicle exhaust and non-exhaust processes play a significant role in environmental pollution, as they are a source of the finest particulate matter. Emissions from non-exhaust processes include wear-products of brakes, tires, automotive hardware, road surface, and traffic signs, but still are paid little attention to. Automotive friction composites for brake pads are composite materials which may consist of potentially hazardous materials and there is a lack of information regarding the potential influence of the brake wear debris (BWD) on the environment, especially on human health. Thus, we focused our study on the genotoxicity of the airborne fraction of BWDmore » using a brake pad model representing an average low-metallic formulation available in the EU market. BWD was generated in the laboratory by a full-scale brake dynamometer and characterized by Raman microspectroscopy, scanning electron microscopy, and transmission electron microscopy showing that it contains nano-sized crystalline metal-based particles. Genotoxicity tested in human lymphocytes in different testing conditions showed an increase in frequencies of micronucleated binucleated cells (MNBNCs) exposed for 48 h to BWD nanoparticles (NPs) (with 10% of foetal calf serum in culture medium) compared with lymphocytes exposed to medium alone, statistically significant only at the concentration 3 µg/cm{sup 2} (p=0.032). - Highlights: • BWD was characterized by Raman microspectroscopy, scanning electron microscopy, and transmission electron microscopy. • Our result showed that BWD contains crystalline metal NPs. • Two different protocols for CBMN assay were used to study of genotoxicity of BWD. • We found significantly increased frequency of MNBNCs after 48 h exposure of BWD (with 10% of foetal calf serum in culture media) at the concentration 3 µg/cm{sup 2}.« less

  14. Locally distributed ground deformation in an area of potential phreatic eruption, Midagahara volcano, Japan, detected by single-look-based InSAR time series analysis

    NASA Astrophysics Data System (ADS)

    Kobayashi, Tomokazu

    2018-05-01

    Although it is difficult to monitor the spatial extent and temporal evolution of local and small-magnitude ground inflation, this information is vital to assess the potential for phreatic eruption. Herein, we demonstrate the detection of locally distributed ground deformation preceding the enhancement of geothermal activity in the Midagahara volcano, Japan, through the application of single-look-based interferometric synthetic aperture radar analysis. In the Jigoku-dani geothermal area, the ground deformation proceeded at a low speed of 4 cm/year at most with a spatial extent of 500 m in the east-west direction and 250 m in the north-south direction. The deformation can be recognized to progress from 2007, at the latest, to 2010, after which the geothermal activity increased, with the collapse of sulfur towers and the appearance of active fumaroles and boiling water on the ground surface. The most deformed area corresponds to the geothermal area with the highest activity observed on the ground surface. Assuming a sill opening model, the deformation source is estimated to be located at a depth of 50 m from the surface with a speed of 7 cm/year at most, which is consistent with the depth of the highly conductive medium inferred from magnetotelluric analyses. This may suggest that volcanic fluid and/or heat was injected into the fluid-rich medium from depth and caused the ground inflation. Our results demonstrate that high-spatial-resolution deformation data can be an effective tool to monitor subsurface pressure conditions with pinpoint spatial accuracy during the build-up to phreatic eruptions.

  15. In vitro propagation of the medicinal plant Ziziphora tenuior L. and evaluation of its antioxidant activity

    PubMed Central

    Dakah, Abdulkarim; Zaid, Salim; Suleiman, Mohamad; Abbas, Sami; Wink, Michael

    2014-01-01

    Ziziphora tenuior L. (Lamiaceae) is an aromatic herb used for its medicinal values against fungi, bacteria. Micropropagation can be used for large-scale multiplication of essential oil producing plants thus avoiding an overexploitation of natural resources. This work aims to develop a reliable protocol for the in vitro propagation of Z. tenuior, and to compare the antioxidant activity between in vitro propagated and wild plants. The explants were sterilized and cultured on MS medium containing different concentrations of growth regulators naphthalene acetic acid (NAA) or indole-3-butyric acid (IBA) with 0.5 mg/L of kinetin (Kin) callus formation was 70.2% after 45 days of incubation in dark on medium supplemented with 1.5 mg/L of NAA. After one month of callus culture on medium supplemented with 2 mg/L BA the shoot number was 5.12 and for the multiplication stage. The shoot number was 4.21 and length was 6.17 cm on medium supplemented with 1 mg/L Kin + 0.1 mg/L NAA. DPPH• reagent was used to test the antioxidant activity. The aqueous and methanol extracts of in vitro plants which were treated with 1.5 and 1 mg/L of kin plus 0.1 mg/L of NAA showed a strong DPPH• scavenging activity where IC50 was 0.307 and 0.369 mg/ml, respectively, while the IC50 of aqueous and methanol extracts of wild plants was 0.516 and 9.229 mg/ml, respectively. Our results suggested that plant growth regulators and in vitro culture conditions increased the antioxidant activity. PMID:25183942

  16. In vitro propagation of the medicinal plant Ziziphora tenuior L. and evaluation of its antioxidant activity.

    PubMed

    Dakah, Abdulkarim; Zaid, Salim; Suleiman, Mohamad; Abbas, Sami; Wink, Michael

    2014-09-01

    Ziziphora tenuior L. (Lamiaceae) is an aromatic herb used for its medicinal values against fungi, bacteria. Micropropagation can be used for large-scale multiplication of essential oil producing plants thus avoiding an overexploitation of natural resources. This work aims to develop a reliable protocol for the in vitro propagation of Z. tenuior, and to compare the antioxidant activity between in vitro propagated and wild plants. The explants were sterilized and cultured on MS medium containing different concentrations of growth regulators naphthalene acetic acid (NAA) or indole-3-butyric acid (IBA) with 0.5 mg/L of kinetin (Kin) callus formation was 70.2% after 45 days of incubation in dark on medium supplemented with 1.5 mg/L of NAA. After one month of callus culture on medium supplemented with 2 mg/L BA the shoot number was 5.12 and for the multiplication stage. The shoot number was 4.21 and length was 6.17 cm on medium supplemented with 1 mg/L Kin + 0.1 mg/L NAA. DPPH• reagent was used to test the antioxidant activity. The aqueous and methanol extracts of in vitro plants which were treated with 1.5 and 1 mg/L of kin plus 0.1 mg/L of NAA showed a strong DPPH• scavenging activity where IC50 was 0.307 and 0.369 mg/ml, respectively, while the IC50 of aqueous and methanol extracts of wild plants was 0.516 and 9.229 mg/ml, respectively. Our results suggested that plant growth regulators and in vitro culture conditions increased the antioxidant activity.

  17. Experimental Setup to Assess Blast and Penetration-Induced Secondary Debris in a Military Operations in Urban Terrain (MOUT) Environment

    DTIC Science & Technology

    2015-11-01

    Fig. 4 Collection pack with gelatin soap and synthetic gelatin ........................4 Fig. 5 Gelatin soap ...designed to hold 2 synthetic gelatin blocks or a combination of gelatin soap and synthetic gelatin for use as the collection medium (Fig. 4). Shelves...were constructed to support the synthetic gelatin and gelatin soap with dimensions of 25.4 cm wide × 25.4 cm deep. Each shelf was attached to the

  18. Packed Bed Bioreactor for the Isolation and Expansion of Placental-Derived Mesenchymal Stromal Cells

    PubMed Central

    Osiecki, Michael J.; Michl, Thomas D.; Kul Babur, Betul; Kabiri, Mahboubeh; Atkinson, Kerry; Lott, William B.; Griesser, Hans J.; Doran, Michael R.

    2015-01-01

    Large numbers of Mesenchymal stem/stromal cells (MSCs) are required for clinical relevant doses to treat a number of diseases. To economically manufacture these MSCs, an automated bioreactor system will be required. Herein we describe the development of a scalable closed-system, packed bed bioreactor suitable for large-scale MSCs expansion. The packed bed was formed from fused polystyrene pellets that were air plasma treated to endow them with a surface chemistry similar to traditional tissue culture plastic. The packed bed was encased within a gas permeable shell to decouple the medium nutrient supply and gas exchange. This enabled a significant reduction in medium flow rates, thus reducing shear and even facilitating single pass medium exchange. The system was optimised in a small-scale bioreactor format (160 cm2) with murine-derived green fluorescent protein-expressing MSCs, and then scaled-up to a 2800 cm2 format. We demonstrated that placental derived MSCs could be isolated directly within the bioreactor and subsequently expanded. Our results demonstrate that the closed system large-scale packed bed bioreactor is an effective and scalable tool for large-scale isolation and expansion of MSCs. PMID:26660475

  19. Presentation of a complex permittivity-meter with applications for sensing the moisture and salinity of a porous media.

    PubMed

    Chavanne, Xavier; Frangi, Jean-Pierre

    2014-08-26

    This paper describes a sensor dedicated to measuring the vertical profile of the complex permittivity and the temperature of any medium in which sensor electrodes are inserted. Potential applications are the estimate of the humidity and salinity in a porous medium, such as a soil. It consists of vertically-stacked capacitors along two conductive parallel cylinders of 5 cm in diameter and at a 10-cm distance to scan a significant volume of the medium (~1 L). It measures their admittances owing to a self-balanced impedance bridge operating at a frequency in the range of 1-20 MHz, possibly 30 MHz. Thanks to accurate design and electronic circuit theory-based modeling, the determination of the admittances takes into account all distortions due to lead and bridge electromagnetic effects inside the sensor when working at high frequencies. Calibration procedures and uncertainties are presented. The article also describes developments to make the present sensor autonomous on digital acquisition, basic data treatment and energy, as well as able to transfer stored data by a radio link. These steps in progress are prerequisites for a wireless network of sensors.

  20. Packed Bed Bioreactor for the Isolation and Expansion of Placental-Derived Mesenchymal Stromal Cells.

    PubMed

    Osiecki, Michael J; Michl, Thomas D; Kul Babur, Betul; Kabiri, Mahboubeh; Atkinson, Kerry; Lott, William B; Griesser, Hans J; Doran, Michael R

    2015-01-01

    Large numbers of Mesenchymal stem/stromal cells (MSCs) are required for clinical relevant doses to treat a number of diseases. To economically manufacture these MSCs, an automated bioreactor system will be required. Herein we describe the development of a scalable closed-system, packed bed bioreactor suitable for large-scale MSCs expansion. The packed bed was formed from fused polystyrene pellets that were air plasma treated to endow them with a surface chemistry similar to traditional tissue culture plastic. The packed bed was encased within a gas permeable shell to decouple the medium nutrient supply and gas exchange. This enabled a significant reduction in medium flow rates, thus reducing shear and even facilitating single pass medium exchange. The system was optimised in a small-scale bioreactor format (160 cm2) with murine-derived green fluorescent protein-expressing MSCs, and then scaled-up to a 2800 cm2 format. We demonstrated that placental derived MSCs could be isolated directly within the bioreactor and subsequently expanded. Our results demonstrate that the closed system large-scale packed bed bioreactor is an effective and scalable tool for large-scale isolation and expansion of MSCs.

  1. Presentation of a Complex Permittivity-Meter with Applications for Sensing the Moisture and Salinity of a Porous Media

    PubMed Central

    Chavanne, Xavier; Frangi, Jean-Pierre

    2014-01-01

    This paper describes a sensor dedicated to measuring the vertical profile of the complex permittivity and the temperature of any medium in which sensor electrodes are inserted. Potential applications are the estimate of the humidity and salinity in a porous medium, such as a soil. It consists of vertically-stacked capacitors along two conductive parallel cylinders of 5 cm in diameter and at a 10-cm distance to scan a significant volume of the medium (∼1 L). It measures their admittances owing to a self-balanced impedance bridge operating at a frequency in the range of 1–20 MHz, possibly 30 MHz. Thanks to accurate design and electronic circuit theory-based modeling, the determination of the admittances takes into account all distortions due to lead and bridge electromagnetic effects inside the sensor when working at high frequencies. Calibration procedures and uncertainties are presented. The article also describes developments to make the present sensor autonomous on digital acquisition, basic data treatment and energy, as well as able to transfer stored data by a radio link. These steps in progress are prerequisites for a wireless network of sensors. PMID:25162233

  2. Effect of adenine sulphate interaction on growth and development of shoot regeneration and inhibition of shoot tip necrosis under in vitro condition in adult Syzygium cumini L.--a multipurpose tree.

    PubMed

    Naaz, Afshan; Shahzad, Anwar; Anis, Mohammad

    2014-05-01

    An efficient method for cloning Syzygium cumini (above 40 years old) through mature nodal segments has been successfully developed and that could be exploited for large-scale production of this valuable multipurpose tree. Nodal segments from mature tree were taken as explants and cultured on MS basal medium with different cytokinins (BA, Kin, AdS). The application of BA proved to be the best responsive cytokinin for the induction of shoot buds and shoots, but the proliferated shoots exhibited slower and stunted growth accompanied with abscission of leaves and shoot tip necrosis (STN). The problem of leaf abscission and STN was considerably reduced by the application of an adjuvant, adenine sulphate (AdS) in the optimal medium which led to the production of a maximum of 14 shoots. Further improvement in shoot bud regeneration and improved growth pattern of the regenerating tissue was obtained on the media comprised of MS + BA (10 μM) + GA3 (2.5 μM). A total number of 15 shoots with mean shoot length of 5.9 cm was obtained. The healthy elongated shoots were then rooted on MS basal augmented with NAA (5 μM). The plantlets obtained were healthy and were successfully acclimatized and transferred under field condition with 70 % survival rate.

  3. The distribution of interstellar dust in the solar neighborhood

    NASA Technical Reports Server (NTRS)

    Gaustad, John E.; Van Buren, Dave

    1993-01-01

    We surveyed the IRAS data base at the positions of the 1808 O6-B9.5 stars in The Bright Star Catalog for extended objects with excess emission at 60 microns, indicating the presence of interstellar dust at the location of the star. Within 400 pc the filling factor of the interstellar medium, for dust clouds with a density greater than 0.5/cu cm is 14.6 + or - 2.4%. Above a density of 1.0/cu cm, the density distribution function appears to follow a power law index - 1.25. When the dust clouds are mapped onto the galactic plane, the sun appears to be located in a low-density region of the interstellar medium of width about 60 pc extending at least 500 pc in the direction of longitudes 80 deg - 260 deg, a feature we call the 'local trough'.

  4. Evidence of reversible bradycardia and arrhythmias caused by immunogenic proteins secreted by T. cruzi in isolated rat hearts.

    PubMed

    Rodríguez-Angulo, Héctor O; Toro-Mendoza, Jhoan; Marques, Juan A; Concepción, Juan L; Bonfante-Cabarcas, Rafael; Higuerey, Yoliver; Thomas, Luz E; Balzano-Nogueira, Leandro; López, José R; Mijares, Alfredo

    2015-02-01

    Chagas cardiomyopathy, caused by the protozoan Trypanosoma cruzi, is characterized by alterations in intracellular ion, heart failure and arrhythmias. Arrhythmias have been related to sudden death, even in asymptomatic patients, and their molecular mechanisms have not been fully elucidated. The aim of this study is to demonstrate the effect of proteins secreted by T. cruzi on healthy, isolated beating rat heart model under a non-damage-inducing protocol. We established a non-damage-inducing recirculation-reoxygenation model where ultrafiltrate fractions of conditioned medium control or conditioned infected medium were perfused at a standard flow rate and under partial oxygenation. Western blotting with chagasic patient serum was performed to determine the antigenicity of the conditioned infected medium fractions. We observed bradycardia, ventricular fibrillation and complete atrioventricular block in hearts during perfusion with >50 kDa conditioned infected culture medium. The preincubation of conditioned infected medium with chagasic serum abolished the bradycardia and arrhythmias. The proteins present in the conditioned infected culture medium of >50 kDa fractions were recognized by the chagasic patient sera associated with arrhythmias. These results suggest that proteins secreted by T. cruzi are involved in Chagas disease arrhythmias and may be a potential biomarker in chagasic patients.

  5. Evidence of Reversible Bradycardia and Arrhythmias Caused by Immunogenic Proteins Secreted by T. cruzi in Isolated Rat Hearts

    PubMed Central

    Rodríguez-Angulo, Héctor O.; Toro-Mendoza, Jhoan; Marques, Juan A.; Concepción, Juan L.; Bonfante-Cabarcas, Rafael; Higuerey, Yoliver; Thomas, Luz E.; Balzano-Nogueira, Leandro; López, José R.; Mijares, Alfredo

    2015-01-01

    Rationale Chagas cardiomyopathy, caused by the protozoan Trypanosoma cruzi, is characterized by alterations in intracellular ion, heart failure and arrhythmias. Arrhythmias have been related to sudden death, even in asymptomatic patients, and their molecular mechanisms have not been fully elucidated. Objective The aim of this study is to demonstrate the effect of proteins secreted by T. cruzi on healthy, isolated beating rat heart model under a non-damage-inducing protocol. Methods and Results We established a non-damage-inducing recirculation-reoxygenation model where ultrafiltrate fractions of conditioned medium control or conditioned infected medium were perfused at a standard flow rate and under partial oxygenation. Western blotting with chagasic patient serum was performed to determine the antigenicity of the conditioned infected medium fractions. We observed bradycardia, ventricular fibrillation and complete atrioventricular block in hearts during perfusion with >50 kDa conditioned infected culture medium. The preincubation of conditioned infected medium with chagasic serum abolished the bradycardia and arrhythmias. The proteins present in the conditioned infected culture medium of >50 kDa fractions were recognized by the chagasic patient sera associated with arrhythmias. Conclusions These results suggest that proteins secreted by T. cruzi are involved in Chagas disease arrhythmias and may be a potential biomarker in chagasic patients. PMID:25647069

  6. Tumor necrosis factor-inducible gene 6 promotes liver regeneration in mice with acute liver injury.

    PubMed

    Wang, Sihyung; Lee, Ji-Seon; Hyun, Jeongeun; Kim, Jieun; Kim, Seung U; Cha, Hyuk-Jin; Jung, Youngmi

    2015-03-11

    Tumor necrosis factor-inducible gene 6 protein (TSG-6), one of the cytokines released by human mesenchymal stem/stromal cells (hMSC), has an anti-inflammatory effect and alleviates several pathological conditions; however, the hepatoprotective potential of TSG-6 remains unclear. We investigated whether TSG-6 promoted liver regeneration in acute liver failure. The immortalized hMSC (B10) constitutively over-expressing TSG-6 or empty plasmid (NC: Negative Control) were established, and either TSG-6 or NC-conditioned medium (CM) was intraperitoneally injected into mice with acute liver damage caused by CCl4. Mice were sacrificed at 3 days post-CM treatment. Higher expression and the immunosuppressive activity of TSG-6 were observed in CM from TSG-6-hMSC. The obvious histomorphological liver injury and increased level of liver enzymes were shown in CCl4-treated mice with or without NC-CM, whereas those observations were markedly ameliorated in TSG-6-CM-treated mice with CCl4. Ki67-positive hepatocytic cells were accumulated in the liver of the CCl4+TSG-6 group. RNA analysis showed the decrease in both of inflammation markers, tnfα, il-1β, cxcl1 and cxcl2, and fibrotic markers, tgf-β1, α-sma and collagen α1, in the CCl4+TSG-6 group, compared to the CCl4 or the CCl4+NC group. Protein analysis confirmed the lower expression of TGF-β1 and α-SMA in the CCl4+TSG-6 than the CCl4 or the CCl4+NC group. Immunostaining for α-SMA also revealed the accumulation of the activated hepatic stellate cells in the livers of mice in the CCl4 and CCl4+NC groups, but not in the livers of mice from the CCl4+TSG-6 group. The cultured LX2 cells, human hepatic stellate cell line, in TSG-6-CM showed the reduced expression of fibrotic markers, tgf-β1, vimentin and collagen α1, whereas the addition of the TSG-6 antibody neutralized the inhibitory effect of TSG-6 on the activation of LX2 cells. In addition, cytoplasmic lipid drops, the marker of inactivated hepatic stellate cell, were detected in TSG-6-CM-cultured LX2 cells, only. The suppressed TSG-6 activity by TSG-6 antibody attenuated the restoration process in livers of TSG-6-CM-treated mice with CCl4. These results demonstrated that TSG-6 contributed to the liver regeneration by suppressing the activation of hepatic stellate cells in CCl4-treated mice, suggesting the therapeutic potential of TSG-6 for acute liver failure.

  7. Impact of Nutrient Restriction on the Structure of Listeria monocytogenes Biofilm Grown in a Microfluidic System.

    PubMed

    Cherifi, Tamazight; Jacques, Mario; Quessy, Sylvain; Fravalo, Philippe

    2017-01-01

    Biofilm formation by the pathogen Listeria monocytogenes is a major concern in food industries. The aim of this work was to elucidate the effect of nutrient limitation on both biofilm architecture and on the viability of the bacteria in microfluidic growth conditions. Biofilm formation by two L. monocytogenes strains was performed in a rich medium (BHI) and in a 10-fold diluted BHI (BHI/10) at 30°C for 24 h by using both static conditions and the microfluidic system Bioflux. In dynamic conditions, biofilms grown in rich and poor medium showed significant differences as well in structure and in the resulting biovolume. In BHI/10, biofilm was organized in a knitted network where cells formed long chains, whereas in the rich medium, the observed structure was homogeneous cellular multilayers. Biofilm biovolume production in BHI/10 was significantly higher than in BHI in these dynamic conditions. Interestingly, biovolume of dead cells in biofilms formed under limited nutrient conditions (BHI/10) was significantly higher than in biofilms formed in the BHI medium. In the other hand, in static conditions, biofilm is organized in a multilayer cells and dispersed cells in a rich medium BHI and poor medium BHI/10 respectively. There was significantly more biomass in the rich medium compared to BHI/10 but no difference was noted in the dead/damaged subpopulation showing how L. monocytogenes biofilm could be affected by the growth conditions. This work demonstrated that nutrient concentration affects biofilm structure and the proportion of dead cells in biofilms under microfluidic condition. Our study also showed that limited nutrients play an important role in the structural stability of L. monocytogenes biofilm by enhancing cell death and liberating extracellular DNA.

  8. Impact of Nutrient Restriction on the Structure of Listeria monocytogenes Biofilm Grown in a Microfluidic System

    PubMed Central

    Cherifi, Tamazight; Jacques, Mario; Quessy, Sylvain; Fravalo, Philippe

    2017-01-01

    Biofilm formation by the pathogen Listeria monocytogenes is a major concern in food industries. The aim of this work was to elucidate the effect of nutrient limitation on both biofilm architecture and on the viability of the bacteria in microfluidic growth conditions. Biofilm formation by two L. monocytogenes strains was performed in a rich medium (BHI) and in a 10-fold diluted BHI (BHI/10) at 30°C for 24 h by using both static conditions and the microfluidic system Bioflux. In dynamic conditions, biofilms grown in rich and poor medium showed significant differences as well in structure and in the resulting biovolume. In BHI/10, biofilm was organized in a knitted network where cells formed long chains, whereas in the rich medium, the observed structure was homogeneous cellular multilayers. Biofilm biovolume production in BHI/10 was significantly higher than in BHI in these dynamic conditions. Interestingly, biovolume of dead cells in biofilms formed under limited nutrient conditions (BHI/10) was significantly higher than in biofilms formed in the BHI medium. In the other hand, in static conditions, biofilm is organized in a multilayer cells and dispersed cells in a rich medium BHI and poor medium BHI/10 respectively. There was significantly more biomass in the rich medium compared to BHI/10 but no difference was noted in the dead/damaged subpopulation showing how L. monocytogenes biofilm could be affected by the growth conditions. This work demonstrated that nutrient concentration affects biofilm structure and the proportion of dead cells in biofilms under microfluidic condition. Our study also showed that limited nutrients play an important role in the structural stability of L. monocytogenes biofilm by enhancing cell death and liberating extracellular DNA. PMID:28567031

  9. A survey of interstellar HI from L alpha absorption measurements 2

    NASA Technical Reports Server (NTRS)

    Bohlin, R. C.; Savage, B. D.; Drake, J. F.

    1977-01-01

    The Copernicus satellite surveyed the spectral region near L alpha to obtain column densities of interstellar HI toward 100 stars. The distance to 10 stars exceeds 2 kpc and 34 stars lie beyond 1 kpc. Stars with color excess E(B-V) up to 0.5 mag are observed. The value of the mean ratio of total neutral hydrogen to color excess was found to equal 5.8 x 10 to the 21st power atoms per (sq cm x mag). For stars with accurate E(B-V), the deviations from this mean are generally less than a factor of 1.5. A notable exception is the dark cloud star, rho Oph. A reduction in visual reddening efficiency for the grains that are larger than normal in the rho Oph dark cloud probably explains this result. The conversion of atomic hydrogen into molecular form in dense clouds was observed in the gas to E(B-V) correlation plots. The best estimate for the mean total gas density for clouds and the intercloud medium, as a whole, in the solar neighborhood and in the plane of the galaxy is 1.15 atoms per cu. cm; those for the atomic gas and molecular gas alone are 0.86 atoms per cu cm and 0.143 molecules per cu cm respectively. For the intercloud medium, where molecular hydrogen is a negligible fraction of the total gas, atomic gas density was found to equal 0.16 atoms per cu cm with a Gaussian scale height perpendicular to the plane of about 350 pc, as derived from high latitude stars.

  10. An efficient and reproducible method for in vitro clonal multiplication of Rauvolfia tetraphylla L. and evaluation of genetic stability using DNA-based markers.

    PubMed

    Faisal, Mohammad; Alatar, Abdulrahman A; Ahmad, Naseem; Anis, Mohammad; Hegazy, Ahmad K

    2012-12-01

    An efficient protocol is described for the rapid in vitro clonal propagation of an endangered medicinal plant, Rauvolfia tetraphylla L., through high frequency shoot induction from nodal explants collected from young shoots of a field grown plant. Effects of growth regulators [6-benzyladenine (BA), kinetin (Kin) 2iP, or α-naphthalene acetic acid (NAA)], carbohydrates, different medium [Murashige and Skoog (MS), Woody Plant Medium (WPM), Gamborg medium (B5), Linsmier and Skoog medium (LS)], and various pH levels on in vitro morphogenesis were investigated. The highest frequency of shoot regeneration (90 %) and maximum number of shoot (35.4 ± 2.3) per explant were observed on WPM medium supplemented with 7.5 μM BA, 2.5 μM NAA, and 30 g/l sucrose at pH 5.8. Well-developed shoots, 4-5 cm in length, were successfully rooted ex vitro at 90 % by a 30-min pulse treatment with 150 μM IBA prior to their transfer in planting substrates. The survival rate of transplantation reached 90 % when transferred to field condition. Genetic stability of micropropagated plantlets was assessed and compared with mother plant using Random Amplified Polymorphic DNA and Inter Simple Sequence Repeats markers. No variation was observed in DNA fingerprinting patterns among the micropropagated plants, which were similar to that of the donor plant illustrating their genetic uniformity and clonal fidelity. This confirms that clonal propagation of this plant using axillary shoot buds can be used for commercial exploitation of the selected genotype where a high degree of fidelity is an essential prerequisite. The work contributed to a better in vitro regeneration and clonal mass multiplication of R. tetraphylla and to develop a strategy for the germplasm conservation of this endangered medicinal plant.

  11. Mesenchymal stem cell-conditioned medium triggers neuroinflammation and reactive species generation in organotypic cultures of rat hippocampus.

    PubMed

    Horn, Ana Paula; Bernardi, Andressa; Luiz Frozza, Rudimar; Grudzinski, Patrícia Bencke; Hoppe, Juliana Bender; de Souza, Luiz Fernando; Chagastelles, Pedro; de Souza Wyse, Angela Terezinha; Bernard, Elena Aida; Battastini, Ana Maria Oliveira; Campos, Maria Martha; Lenz, Guido; Nardi, Nance Beyer; Salbego, Christianne

    2011-07-01

    Cell therapy using bone marrow-derived mesenchymal stem cells (MSCs) seems to be a new alternative for the treatment of neurodegenerative diseases. Despite several promising results with their use, possible side effects are still unknown. In a previous work, we have shown that MSC-conditioned medium is toxic to hippocampal slice cultures and aggravates cell death induced by oxygen and glucose deprivation. In this work, we investigated whether the inflammatory response and/or reactive species formation could be involved in that toxicity. Rat organotypic hippocampal cultures were exposed for 24 h to conditioned medium from MSCs isolated from rat bone marrow. A marked glial activation was observed after exposure of cultures to MSC-conditioned medium, as evidenced by glial fibrillary acid protein (GFAP) and isolectin B(4) increase. Tumor necrosis factor-α and interleukin-6 levels were increased in the culture medium, and 2,7-dihydrodichlorofluorescein diacetate oxidation (indicating reactive species generation) and inducible nitric oxide synthase (iNOS) immunocontent were also higher after exposure of cultures to MSC-conditioned medium. Antioxidants (ascorbic acid and TROLOX(®)), N(ω)-nitro-l-arginine methyl ester hydrochloride, and anti-inflammatory drugs (indomethacin and dexamethasone) reduced cell death in hippocampal organotypic cultures after their exposure to MSC-conditioned medium. The results obtained here suggest that MSC-secreted factors trigger reactive species generation and neuroinflammation in organotypic cultures of hippocampus, introducing a note of caution in the use of these cells for neurological application.

  12. Vertical electric field stimulation of neural cells on porous amorphous carbon electrodes

    NASA Astrophysics Data System (ADS)

    Jain, Shilpee; Sharma, Ashutosh; Basu, Bikramjit

    2014-03-01

    We demonstrate the efficacy of amorphous macroporous carbon substrates as electrodes to stimulate neuronal cell proliferation in presence of external electric field. The electric field was applied perpendicular to carbon electrode, while growing mouse neuroblastoma (N2a) cells in vitro. The placement of the second electrode outside of the cell culture medium allows the investigation of cell response to electric field without the concurrent complexities of submerged electrodes such as potentially toxic electrode reactions, electro-kinetic flows and charge transfer (electrical current) in the cell medium. The macroporous carbon electrodes are uniquely characterized by a higher specific charge storage capacity (0.2 mC/cm2) and low impedance (3.3 k Ω at 1 kHz). When a uniform or a gradient electric field was applied perpendicular to the amorphous carbon substrate, it was found that the N2a cell viability and neurite length were higher at low electric field strengths (<= 2.5 V/cm) compared to that measured without an applied field (0 V/cm). Overall, the results of the present study unambiguously establish the uniform/gradient vertical electric field based culture protocol to stimulate neurite outgrowth and viability of nerve cells.

  13. Phototoxicity of argon laser irradiation on biofilms of Porphyromonas and Prevotella species.

    PubMed

    Henry, C A; Dyer, B; Wagner, M; Judy, M; Matthews, J L

    1996-07-01

    Species of Prevotella (Pr.) and Porphyromonas (Po.) and other microorganisms were cultivated as biofilms on agar medium and examined for their susceptibility to argon laser irradiation (continuous mode; wavelengths, 488-514 nm; fluences, 20-200 J cm(-2)). Fluences of 35 to 80 J cm(-2) inhibited biofilm growth in Po. endodontalis, Po. gingivalis, Pr. denticola, Pr. intermedia, Pr. melaninogenica and Pr. nigrescens. A fluence of 70 J cm(-2) did not affect biofilm growth in species of Bacillus, Candida, Enterobacter, Proteus, Pseudomonas, Staphylococcus and Streptococcus. The phototoxic effects of argon laser irradiation against Prevotella and Porphyromonas species were: (1) caused by the radiation alone; (2) modified by biofilm age; (3) dependent on the presence of atmospheric oxygen; (4) influenced by medium supplements of hemin, hemoglobin and blood; (5) greater when compared with other microbial species; (6) demonstrated without augmentation with an exogenous photosensitizer; and (7) apparently unrelated to the protoporphyrin content of the cells. Overall, these in vitro findings suggest that low doses of argon laser radiation may be effective in the treatment and/or prevention of clinical infections caused by biofilm-associated species of Prevotella or Porphyromonas.

  14. Chemistry in Magnetohydrodynamic Shock Waves in Diffuse Molecular Clouds

    NASA Astrophysics Data System (ADS)

    Peimbert, Antonio

    1998-09-01

    Absorption observations of the CH+ molecule with column densities of up to 1014 cm-2 in diffuse molecular clouds in many lines of sight are reviewed, and compared to the reddening and to abundances and velocity shifts of molecules like CH. Special attention is placed on the observations of the line of sight towards ς Ophiuchi where high quality observations of many chemical species are available. The problem of the required CH+ is described, and many formation mechanisms from the literature are reviewed, finding that none of them is particularly apt at describing the observations towards ς-Oph. Two fluid J-type shock models are studied as an alternative. The necessary conditions for their formation are discussed, and it is shown how they are expected to be present widely in the interstellar medium. Plane parallel numerical integrations, for the particular case in which the magnetic field is perpendicular to the shock velocity, are employed to study the region of phase-space of initial conditions that will produce 2 fluid shocks. A chemical network is developed and formation of key molecules like CH+, CH and OH, along with the excited roto-vibrational levels of H2, are studied under the shock dynamics. These models are then compared to the observations of the different lines of sight, showing they are capable of reproducing the features of the observations towards most of those clouds. An attempt to model the line of sight towards ς-Oph is done, finding that a shock with a shock speed vs = 9.0km/s going through a cloud with a density of nH = 14cm-3 with a magnetic field of B = 4.7μG does a reasonable job at satisfying most of the observations with the exception of the highest rotational excited states of molecular hydrogen for which observations are available. There is a small family of solutions capable of explaining the observed results which make specific predictions for the velocity profiles of the H2 lines of various excited levels. New observations with the Interstellar Medium Absorption Profile Spectrograph (IMAPS) camera would be useful in confirming or rejecting these models.

  15. GPER1-mediated IGFBP-1 induction modulates IGF-1-dependent signaling in tamoxifen-treated breast cancer cells.

    PubMed

    Vaziri-Gohar, Ali; Houston, Kevin D

    2016-02-15

    Tamoxifen, a selective estrogen receptor modulator, is a commonly prescribed adjuvant therapy for estrogen receptor-α (ERα)-positive breast cancer patients. To determine if extracellular factors contribute to the modulation of IGF-1 signaling after tamoxifen treatment, MCF-7 cells were treated with IGF-1 in conditioned medium (CM) obtained from 4-OHT-treated MCF-7 cells and the accumulation of phospho-Akt (S473) was measured. CM inhibited IGF-1-dependent cell signaling and suggesting the involvement of extracellular factors (ie. IGFBPs). A significant increase in IGFBP-1 mRNA and extracellular IGFBP-1 protein was observed in 4-OHT-treated MCF-7 cells. Knockdown experiments demonstrated that both GPER1 and CREB mediate IGFBP-1 induction. Furthermore, experiments showed that 4-OHT-dependent IGFBP-1 transcription is downstream of GPER1-activation in breast cancer cells. Additionally, neutralization and knockdown experiments demonstrated a role for IGFBP-1 in the observed inhibition of IGF-1 signaling. These results suggested that 4-OHT inhibits IGF-1 signaling via GPER1 and CREB mediated extracellular IGFBP-1 accumulation in breast cancer cells. Copyright © 2015 The Authors. Published by Elsevier Ireland Ltd.. All rights reserved.

  16. Fast and Accurate Hybrid Stream PCRTMSOLAR Radiative Transfer Model for Reflected Solar Spectrum Simulation in the Cloudy Atmosphere

    NASA Technical Reports Server (NTRS)

    Yang, Qiguang; Liu, Xu; Wu, Wan; Kizer, Susan; Baize, Rosemary R.

    2016-01-01

    A hybrid stream PCRTM-SOLAR model has been proposed for fast and accurate radiative transfer simulation. It calculates the reflected solar (RS) radiances with a fast coarse way and then, with the help of a pre-saved matrix, transforms the results to obtain the desired high accurate RS spectrum. The methodology has been demonstrated with the hybrid stream discrete ordinate (HSDO) radiative transfer (RT) model. The HSDO method calculates the monochromatic radiances using a 4-stream discrete ordinate method, where only a small number of monochromatic radiances are simulated with both 4-stream and a larger N-stream (N = 16) discrete ordinate RT algorithm. The accuracy of the obtained channel radiance is comparable to the result from N-stream moderate resolution atmospheric transmission version 5 (MODTRAN5). The root-mean-square errors are usually less than 5x10(exp -4) mW/sq cm/sr/cm. The computational speed is three to four-orders of magnitude faster than the medium speed correlated-k option MODTRAN5. This method is very efficient to simulate thousands of RS spectra under multi-layer clouds/aerosols and solar radiation conditions for climate change study and numerical weather prediction applications.

  17. Spectrophotometric determination of fluoxetine hydrochloride in bulk and in pharmaceutical formulations.

    PubMed

    Prabhakar, A H; Patel, V B; Giridhar, R

    1999-07-01

    Two new rapid, sensitive and economical spectrophotometric methods are described for the determination of fluoxetine hydrochloride in bulk and in pharmaceutical formulations. Both methods are based on the formation of a yellow ion-pair complex due to the action of methyl orange (MO) and thymol blue (TM) on fluoxetine in acidic (pH 4.0) and basic (pH 8.0) medium, respectively. Under optimised conditions they show an absorption maxima at 433 nm (MO) and 410 nm (TB), with molar absorptivities of 2.12 x 10(-4) and 4.207 x 10(-3) l mol(-1) cm(-1) and Sandell's Sensitivities of 1.64 x 10(-2) and 0.082 microg cm(-2) per 0.001 absorbance unit for MO and TB, respectively. The colour is stable for 5 min after extraction. In both cases Beer's Law is obeyed at 1-20 microg mol(-1) with MO and 4-24 microg mol(-1) with TB. The proposal method was successfully extended to pharmaceutical preparations capsules. The results obtained by both the agreement and E.P. (3rd edition) were in good agreement and statistical comparison by Student's t-test and variance ratio F-test showed no significant difference in the three methods.

  18. Production of Multiple Growth Factors by a Newly Established Human Thyroid Carcinoma Cell Line

    PubMed Central

    Yoshida, Yataro; Ohashi, Kensaku; Sano, Emiko; Kobayashi, Hisataka; Endo, Keigo; Naruto, Masanobu; Nakamura, Toru

    1992-01-01

    A multiple growth factor‐producing tumor cell line (NIM‐1) was newly established from a patient with thyroid cancer and remarkable neutrophilia. NIM‐1 cells also caused severe neutrophilia in nude mice bearing tumors. NIM‐1‐conditioned medium (NIM‐1CM) contained activities that supported not only granulocyte, macrophage and eosinophil colony formation of human bone marrow cells but also the growth of colony‐stimulating factor (CSF)‐dependent cell lines, NFS60‐KX and TF‐1. Northern blot hybridization analysis revealed the constitutive expression of granulocyte‐CSF (G‐CSF), granulocyte/macrophage‐CSF (GM‐CSF) and interleukin(IL)‐6 mRNAs in NIM‐1 cells. Enzyme‐linked immunosorbent assays (ELISA) using NIM‐1CM also confirmed the production of IL‐la and a small amount of IL‐1β besides G‐CSF, GM‐CSF and IL‐6 in NIM‐1 cells. In addition, unexpected production of IL‐11 in NIM‐1 cells was detected by northern blot hybridization analysis and by bioassay using an IL‐11‐dependent cell line. Therefore, NIM‐1 cell line is shown to produce multiple cytokines including potentially megakaryopoietic growth factors such as GM‐CSF, IL‐6 and IL‐11. PMID:1372885

  19. Epac1 increases migration of endothelial cells and melanoma cells via FGF2-mediated paracrine signaling

    PubMed Central

    Baljinnyam, Erdene; Umemura, Masanari; Chuang, Christine; De Lorenzo, Mariana S; Iwatsubo, Mizuka; Chen, Suzie; Goydos, James S; Ishikawa, Yoshihiro; Whitelock, John M; Iwatsubo, Kousaku

    2014-01-01

    Fibroblast growth factor (FGF2) regulates endothelial and melanoma cell migration. The binding of FGF2 to its receptor requires N-sulfated heparan sulfate (HS) glycosamine. We have previously reported that Epac1, an exchange protein activated by cAMP, increases N-sulfation of HS in melanoma. Therefore, we examined whether Epac1 regulates FGF2-mediated cell–cell communication. Conditioned medium (CM) of melanoma cells with abundant expression of Epac1 increased migration of human umbilical endothelial cells (HUVEC) and melanoma cells with poor expression of Epac1. CM-induced increase in migration was inhibited by antagonizing FGF2, by the removal of HS and by the knockdown of Epac1. In addition, knockdown of Epac1 suppressed the binding of FGF2 to FGF receptor in HUVEC, and in vivo angiogenesis in melanoma. Furthermore, knockdown of Epac1 reduced N-sulfation of HS chains attached to perlecan, a major secreted type of HS proteoglycan that mediates the binding of FGF2 to FGF receptor. These data suggested that Epac1 in melanoma cells regulates melanoma progression via the HS–FGF2-mediated cell–cell communication. PMID:24725364

  20. Neural Stem Cells and Its Derivatives as a New Material for Melanin Inhibition

    PubMed Central

    Hwang, Insik

    2017-01-01

    The pigment molecule, melanin, is produced from melanosomes of melanocytes through melanogenesis, which is a complex process involving a combination of chemical and enzymatically catalyzed reactions. The synthesis of melanin is primarily influenced by tyrosinase (TYR), which has attracted interest as a target molecule for the regulation of pigmentation or depigmentation in skin. Thus, direct inhibitors of TYR activity have been sought from various natural and synthetic materials. However, due to issues with these inhibitors, such as weak or permanent ability for depigmentation, allergy, irritant dermatitis and rapid oxidation, in vitro and in vivo, the development of new materials that inhibit melanin production is essential. A conditioned medium (CM) derived from stem cells contains many cell-secreted factors, such as cytokines, chemokines, growth factors and extracellular vesicles including exosomes. In addition, the secreted factors could negatively regulate melanin production through stimulation of a microenvironment of skin tissue in a paracrine manner, which allows the neural stem cell CM to be explored as a new material for skin depigmentation. In this review, we will summarize the current knowledge regulating depigmentation, and discuss the potential of neural stem cells and their derivatives, as a new material for skin depigmentation. PMID:29271951

  1. Influence of repeated screw tightening on bacterial leakage along the implant-abutment interface.

    PubMed

    do Nascimento, Cássio; Pedrazzi, Vinícius; Miani, Paola Kirsten; Moreira, Larissa Daher; de Albuquerque, Rubens Ferreira

    2009-12-01

    Bacterial penetration along the implant-abutment interface as a consequence of abutment screw loosening has been reported in a number of recent studies. The aim of this in vitro study was to investigate the influence of repeated tightening of the abutment screw on leakage of Streptococcus mutans along the interface between implants and pre-machined abutments. Twenty pre-machined abutments with a plastic sleeve were used. The abutment screws were tightened to 32 N cm in group 1 (n=10 - control) and to 32 N cm, loosened and re-tightened with the same torque twice in group 2 (n=10). The assemblies were completely immersed in 5 ml of Tryptic Soy Broth medium inoculated with S. mutans and incubated for 14 days. After this period, contamination of the implant internal threaded chamber was evaluated using the DNA Checkerboard method. Microorganisms were found on the internal surfaces of both groups evaluated. However, bacterial counts in group 2 were significantly higher than that in the control group (P<0.05). These results suggest that bacterial leakage between implants and abutments occurs even under unloaded conditions and at a higher intensity when the abutment screw is tightened and loosened repeatedly.

  2. SU-F-T-365: Clinical Commissioning of the Monaco Treatment Planning System for the Novalis Tx to Deliver VMAT, SRS and SBRT Treatments

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

    Adnani, N

    Purpose: To commission the Monaco Treatment Planning System for the Novalis Tx machine. Methods: The commissioning of Monte-Carlo (MC), Collapsed Cone (CC) and electron Monte-Carlo (eMC) beam models was performed through a series of measurements and calculations in medium and in water. In medium measurements relied Octavius 4D QA system with the 1000 SRS detector array for field sizes less than 4 cm × 4 cm and the 1500 detector array for larger field sizes. Heterogeneity corrections were validated using a custom built phantom. Prior to clinical implementation, an end to end testing of a Prostate and H&N VMAT plansmore » was performed. Results: Using a 0.5% uncertainty and 2 mm grid sizes, Tables I and II summarize the MC validation at 6 MV and 18 MV in both medium and water. Tables III and IV show similar comparisons for CC. Using the custom heterogeneity phantom setup of Figure 1 and IGRT guidance summarized in Figure 2, Table V lists the percent pass rate for a 2%, 2 mm gamma criteria at 6 and 18 MV for both MC and CC. The relationship between MC calculations settings of uncertainty and grid size and the gamma passing rate for a prostate and H&N case is shown in Table VI. Table VII lists the results of the eMC calculations compared to measured data for clinically available applicators and Table VIII for small field cutouts. Conclusion: MU calculations using MC are highly sensitive to uncertainty and grid size settings. The difference can be of the order of several per cents. MC is superior to CC for small fields and when using heterogeneity corrections, regardless of field size, making it more suitable for SRS, SBRT and VMAT deliveries. eMC showed good agreement with measurements down to 2 cm − 2 cm field size.« less

  3. [Spatial variation of soil phosphorus in flooded area of the Yellow River based on GIS and geo-statistical methods: A case study in Zhoukou City, Henan, China.

    PubMed

    Jia, Zhen Yu; Zhang, Jun Hua; Ding, Sheng Yan; Feng, Shu; Xiong, Xiao Bo; Liang, Guo Fu

    2016-04-22

    Soil phosphorus is an important indicator to measure the soil fertility, because the content of soil phosphorus has an important effect on physical and chemical properties of soil, plant growth, and microbial activity in soil. In this study, the soil samples collecting and indoor analysis were conducted in Zhoukou City located in the flooded area of the Yellow River. By using GIS combined with geo-statistics, we tried to analyze the spatial variability and content distribution of soil total phosphorus (TP) and soil available phosphorus (AP) in the study area. Results showed that TP and AP of both soil layers (0-20 cm and 20-40 cm) were rich, and the contents of TP and AP in surface layer (0-20 cm) were higher than in the second layer (20-40 cm). TP and AP of both soil layers exhibited variation at medium level, and AP had varied much higher than TP. TP of both layers showed medium degree of anisotropy which could be well modeled by the Gaussian model. TP in the surface layer showed strong spatial correlation, but that of the second layer had medium spatial correlation. AP of both layers had a weaker scope in anisotropy which could be simulated by linear model, and both soil layers showed weaker spatial correlations. TP of both soil layers showed a slowly rising change from southwest to northeast of the study area, while it gradually declined from northwest to southeast. AP in soil surface layer exhibited an increase tendency firstly and then decrease from southwest to the northeast, while it decreased firstly and then increased from southeast to the northwest. AP in the second soil layer had an opposite change in the southwest to the northeast, while it showed continuously increasing tendency from northwest to the southeast. The contents of TP and AP in the surface layer presented high grades and the second layer of TP belonged to medium grade, but the second layer of AP was in a lower grade. The artificial factors such as land use type, cropping system, irrigation and fertilization were the main factors influencing the distribution and spatial variation of soil phosphorus in this area.

  4. Early results from the Far Infrared Absolute Spectrophotometer (FIRAS)

    NASA Technical Reports Server (NTRS)

    Mather, J. C.; Cheng, E. S.; Shafer, R. A.; Eplee, R. E.; Isaacman, R. B.; Fixsen, D. J.; Read, S. M.; Meyer, S. S.; Weiss, R.; Wright, E. L.

    1991-01-01

    The Far Infrared Absolute Spectrophotometer (FIRAS) on the Cosmic Background Explorer (COBE) mapped 98 percent of the sky, 60 percent of it twice, before the liquid helium coolant was exhausted. The FIRAS covers the frequency region from 1 to 100/cm with a 7 deg angular resolution. The spectral resolution is 0.2/cm for frequencies less than 20/cm and 0.8/cm for higher frequencies. Preliminary results include: a limit on the deviations from a Planck curve of 1 percent of the peak brightness from 1 to 20/cm, a temperature of 2.735 +/- 0.06 K, a limit on the Comptonization parameter y of 0.001, on the chemical potential parameter mu of 0.01, a strong limit on the existence of a hot smooth intergalactic medium, and a confirmation that the dipole anisotropy spectrum is that of a Doppler shifted blackbody.

  5. Macrophage activation by factors released from acetaminophen-injured hepatocytes: Potential role of HMGB1

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

    Dragomir, Ana-Cristina; Laskin, Jeffrey D.; Laskin, Debra L., E-mail: laskin@eohsi.rutgers.edu

    2011-06-15

    Toxic doses of acetaminophen (AA) cause hepatocellular necrosis. Evidence suggests that activated macrophages contribute to the pathogenic process; however, the factors that activate these cells are unknown. In these studies, we assessed the role of mediators released from AA-injured hepatocytes in macrophage activation. Treatment of macrophages with conditioned medium (CM) collected 24 hr after treatment of mouse hepatocytes with 5 mM AA (CM-AA) resulted in increased production of reactive oxygen species (ROS). Macrophage expression of heme oxygenase-1 (HO-1) and catalase mRNA was also upregulated by CM-AA, as well as cyclooxygenase (COX)-2 and 12/15-lipoxygenase (LOX). CM-AA also upregulated expression of themore » proinflammatory chemokines, MIP-1{alpha} and MIP-2. The effects of CM-AA on expression of COX-2, MIP-1{alpha} and MIP-2 were inhibited by blockade of p44/42 MAP kinase, suggesting a biochemical mechanism mediating macrophage activation. Hepatocytes injured by AA were found to release HMGB1, a potent macrophage activator. This was inhibited by pretreatment of hepatocytes with ethyl pyruvate (EP), which blocks HMGB1 release. EP also blocked CM-AA induced ROS production and antioxidant expression, and reduced expression of COX-2, but not MIP-1{alpha} or MIP-2. These findings suggest that HMGB1 released by AA-injured hepatocytes contributes to macrophage activation. This is supported by our observation that expression of the HMGB1 receptor RAGE is upregulated in macrophages in response to CM-AA. These data indicate that AA-injured hepatocytes contribute to the inflammatory environment in the liver through the release of mediators such as HMGB1. Blocking HMGB1/RAGE may be a useful approach to limiting classical macrophage activation and AA-induced hepatotoxicity. - Research Highlights: > These studies analyze macrophage activation by mediators released from acetaminophen-damaged hepatocytes. > Factors released from acetaminophen-injured hepatocytes induce macrophage ROS production and expression of COX-2, chemokines, and RAGE. > Hepatocyte-mediated macrophage activation involves p44/42 MAP kinase signaling. > HMGB1 is released from acetaminophen-injured hepatocytes and contributes to macrophage activation.« less

  6. Evaluation of amniotic mesenchymal cell derivatives on cytokine production in equine alveolar macrophages: an in vitro approach to lung inflammation.

    PubMed

    Zucca, Enrica; Corsini, Emanuela; Galbiati, Valentina; Lange-Consiglio, Anna; Ferrucci, Francesco

    2016-09-20

    Data obtained in both animal models and clinical trials suggest that cell-based therapies represent a potential therapeutic strategy for lung repair and remodeling. Recently, new therapeutic approaches based on the use of stem cell derivatives (e.g., conditioned medium (CM) and microvesicles (MVs)) to regenerate tissues and improve their functions were proposed. The aim of this study was to investigate the immunomodulatory effects of equine amniotic mesenchymal cell derivatives on lipopolysaccharide (LPS)-induced cytokine production in equine alveolar macrophages, which may be beneficial in lung inflammatory disorders such as recurrent airway obstruction (RAO) in horses. RAO shares many features with human asthma, including an increased number of cells expressing mRNA for interleukin (IL)-4 and IL-5 and a decreased expression of IFN-γ in bronchoalveolar lavage fluid (BALF) of affected horses. The release of TNF-α, IL-6, and TGF-β1 at different time points (1, 24, 48, and 72 h) was measured in equine alveolar macrophages stimulated or not with LPS (10 and 100 ng/mL) in the presence or absence of 10 % CM or 50 × 10(6) MVs/mL. Cytokines were measured using commercially available ELISA kits. For multiple comparisons, analysis of variance was used with Tukey post-hoc test. Differences were considered significant at p ≤ 0.05. Significant modulatory effects of CM on LPS-induced TNF-α release at 24 h, and of both CM and MVs on TNF-α release at 48 h were observed. A trend toward a modulatory effect of both CM and MVs on the release of TGF-β and possibly IL-6 was visible over time. Results support the potential use of CM and MVs in lung regenerative medicine, especially in situations in which TGF-β may be detrimental, such as respiratory allergy. Further studies should evaluate the potential clinical applications of CM and MVs in equine lung diseases, such as RAO and other inflammatory disorders.

  7. Absorption by Spinning Dust: A Contaminant for High-redshift 21 cm Observations

    NASA Astrophysics Data System (ADS)

    Draine, B. T.; Miralda-Escudé, Jordi

    2018-05-01

    Spinning dust grains in front of the bright Galactic synchrotron background can produce a weak absorption signal that could affect measurements of high-redshift 21 cm absorption. At frequencies near 80 MHz where the Experiment to Detect the Global EoR Signature (EDGES) has reported 21 cm absorption at z≈ 17, absorption could be produced by interstellar nanoparticles with radii a≈ 50 \\mathringA in the cold interstellar medium (ISM), with rotational temperature T ≈ 50 K. Atmospheric aerosols could contribute additional absorption. The strength of the absorption depends on the abundance of such grains and on their dipole moments, which are uncertain. The breadth of the absorption spectrum of spinning dust limits its possible impact on measurement of a relatively narrow 21 cm absorption feature.

  8. An improved medium for the anaerobic growth of Paracoccus denitrificans Pd1222

    PubMed Central

    Hahnke, Stefanie M.; Moosmann, Philipp; Erb, Tobias J.; Strous, Marc

    2014-01-01

    Paracoccus denitrificans is a well studied model organism with respect to its aerobic and anaerobic respiratory enzymes. However, until now, the growth medium for this organism has not been optimized for anaerobic growth. In particular, the requirements of P. denitrificans for trace elements (TEs) are not well known. In the present study we aimed to improve growth rates of P. denitrificans Pd1222 on a defined medium under anoxic conditions. We designed media containing different combinations of TEs at various concentrations, and tested their performance against previously reported media. Our results suggest that growth rate and yield depend on the availability and concentration of TEs in the medium. A chelated TE solution was more suitable than an acidified TE solution. Highest growth rates were achieved with medium comprising the TEs iron, manganese, molybdenum, copper and zinc ranging from 0.1 to 9 μM. On this medium, P. denitrificans Pd1222 grew with a generation time of 4.4 h under anoxic conditions and 2.8 h under oxic conditions. Diauxic growth was clearly shown with respect to nitrate and nitrite reduction under anoxic conditions. PMID:24550891

  9. An improved medium for the anaerobic growth of Paracoccus denitrificans Pd1222.

    PubMed

    Hahnke, Stefanie M; Moosmann, Philipp; Erb, Tobias J; Strous, Marc

    2014-01-01

    Paracoccus denitrificans is a well studied model organism with respect to its aerobic and anaerobic respiratory enzymes. However, until now, the growth medium for this organism has not been optimized for anaerobic growth. In particular, the requirements of P. denitrificans for trace elements (TEs) are not well known. In the present study we aimed to improve growth rates of P. denitrificans Pd1222 on a defined medium under anoxic conditions. We designed media containing different combinations of TEs at various concentrations, and tested their performance against previously reported media. Our results suggest that growth rate and yield depend on the availability and concentration of TEs in the medium. A chelated TE solution was more suitable than an acidified TE solution. Highest growth rates were achieved with medium comprising the TEs iron, manganese, molybdenum, copper and zinc ranging from 0.1 to 9 μM. On this medium, P. denitrificans Pd1222 grew with a generation time of 4.4 h under anoxic conditions and 2.8 h under oxic conditions. Diauxic growth was clearly shown with respect to nitrate and nitrite reduction under anoxic conditions.

  10. The Role of Paracrine and Autocrine Signaling in the Early Phase of Adipogenic Differentiation of Adipose-derived Stem Cells

    PubMed Central

    Hemmingsen, Mette; Vedel, Søren; Skafte-Pedersen, Peder; Sabourin, David; Collas, Philippe; Bruus, Henrik; Dufva, Martin

    2013-01-01

    Introduction High cell density is known to enhance adipogenic differentiation of mesenchymal stem cells, suggesting secretion of signaling factors or cell-contact-mediated signaling. By employing microfluidic biochip technology, we have been able to separate these two processes and study the secretion pathways. Methods and results Adipogenic differentiation of human adipose-derived stem cells (ASCs) cultured in a microfluidic system was investigated under perfusion conditions with an adipogenic medium or an adipogenic medium supplemented with supernatant from differentiating ASCs (conditioned medium). Conditioned medium increased adipogenic differentiation compared to adipogenic medium with respect to accumulation of lipid-filled vacuoles and gene expression of key adipogenic markers (C/EBPα, C/EBPβ, C/EBPδ, PPARγ, LPL and adiponectin). The positive effects of conditioned medium were observed early in the differentiation process. Conclusions Using different cell densities and microfluidic perfusion cell cultures to suppress the effects of cell-released factors, we have demonstrated the significant role played by auto- or paracrine signaling in adipocyte differentiation. The cell-released factor(s) were shown to act in the recruitment phase of the differentiation process. PMID:23723991

  11. Medium factors on anaerobic production of rhamnolipids by Pseudomonas aeruginosa SG and a simplifying medium for in situ microbial enhanced oil recovery applications.

    PubMed

    Zhao, Feng; Zhou, Jidong; Han, Siqin; Ma, Fang; Zhang, Ying; Zhang, Jie

    2016-04-01

    Aerobic production of rhamnolipid by Pseudomonas aeruginosa was extensively studied. But effect of medium composition on anaerobic production of rhamnolipid by P. aeruginosa was unknown. A simplifying medium facilitating anaerobic production of rhamnolipid is urgently needed for in situ microbial enhanced oil recovery (MEOR). Medium factors affecting anaerobic production of rhamnolipid were investigated using P. aeruginosa SG (Genbank accession number KJ995745). Medium composition for anaerobic production of rhamnolipid by P. aeruginosa is different from that for aerobic production of rhamnolipid. Both hydrophobic substrate and organic nitrogen inhibited rhamnolipid production under anaerobic conditions. Glycerol and nitrate were the best carbon and nitrogen source. The commonly used N limitation under aerobic conditions was not conducive to rhamnolipid production under anaerobic conditions because the initial cell growth demanded enough nitrate for anaerobic respiration. But rhamnolipid was also fast accumulated under nitrogen starvation conditions. Sufficient phosphate was needed for anaerobic production of rhamnolipid. SO4(2-) and Mg(2+) are required for anaerobic production of rhamnolipid. Results will contribute to isolation bacteria strains which can anaerobically produce rhamnolipid and medium optimization for anaerobic production of rhamnolipid. Based on medium optimization by response surface methodology and ions composition of reservoir formation water, a simplifying medium containing 70.3 g/l glycerol, 5.25 g/l NaNO3, 5.49 g/l KH2PO4, 6.9 g/l K2HPO4·3H2O and 0.40 g/l MgSO4 was designed. Using the simplifying medium, 630 mg/l of rhamnolipid was produced by SG, and the anaerobic culture emulsified crude oil to EI24 = 82.5 %. The simplifying medium was promising for in situ MEOR applications.

  12. Burst Speed of Wild Fishes under High-Velocity Flow Conditions Using Stamina Tunnel with Natural Guidance System in River

    NASA Astrophysics Data System (ADS)

    Izumi, Mattashi; Yamamoto, Yasuyuki; Yataya, Kenichi; Kamiyama, Kohhei

    Swimming experiments were conducted on wild fishes in a natural guidance system stamina tunnel (cylindrical pipe) installed in a fishway of a local river under high-velocity flow conditions (tunnel flow velocity : 211 to 279 cm·s-1). In this study, the swimming characteristics of fishes were observed. The results show that (1) the swimming speeds of Tribolodon hakonensis (Japanese dace), Phoxinus lagowshi steindachneri (Japanese fat-minnow), Plecoglossus altivelis (Ayu), and Zacco platypus (Pale chub) were in proportion to their body length under identical water flow velocity conditions; (2) the maximum burst speed of Japanese dace and Japanese fat-minnow (measuring 4 to 6 cm in length) was 262 to 319 cm·s-1 under high flow velocity conditions (225 to 230 cm·s-1), while the maximum burst speed of Ayu and Pale chub (measuring 5 cm to 12 cm in length) was 308 to 355 cm·s-1 under high flow velocity conditions (264 to 273 cm·s-1) ; (3) the 50cm-maximum swimming speed of swimming fishes was 1.07 times faster than the pipe-swimming speed; (4) the faster the flow velocity, the shorter the swimming distance became.

  13. Development and characterization of a compact hand-held gamma probe system, SURGEOGUIDE, based on NEMA NU3-2004 standards

    NASA Astrophysics Data System (ADS)

    Kaviani, S.; Zeraatkar, N.; Sajedi, S.; Gorjizadeh, N.; Farahani, M. H.; Ghafarian, P.; El Fakhri, G.; Sabet, H.; Ay, M. R.

    2016-12-01

    Using an intra-operative gamma probe after periareolar or peritumoral injection of a radiotracer during surgery helps the surgeon to identify the sentinel, or first, nodal site of regional metastasis in clinically node-negative patients. The pathological analysis of this node can have an important influence on the treatment staging in various cancers. This paper reports the design and performance evaluation of a gamma probe recently developed in our department. The detector unit of this system consists of an 8 mm diameter and 10 mm thickness monolithic CsI(Tl) scintillator optically, coupled to a Silicon Photomultiplier (SiPM) with an active area of 6×6 mm2, and a single-hole collimator. The unit is shielded using tungsten. The system can operate in three different modes for Tc-99m, I-131, or F-18 isotopes. The following measurements were carried out to evaluate the performance of the probe: sensitivity in air and scatter medium, spatial resolution in scatter medium, angular resolution in scatter medium, and side and back shielding effectiveness. All experiments have been performed based on the NEMA NU3-2004 standard set up. The measured system sensitivities in air and scatter medium (water) are 1700 cps/MBq and 1770 cps/MBq, respectively, both measured at 3 cm from the collimator. The spatial resolution in the scatter medium is about 45 mm at 3 cm distance from the collimator. Also, the angular resolution of the probe is 74o FWHM. Finally, a shielding effectiveness of 99.5% is measured. The results show that the probe can potentially be used for sentinel lymph node localization during the surgery.

  14. [Stature of Chilean parents and children of different ethnicity and social vulnerability].

    PubMed

    Amigo, H; Erazo, M; Bustos, P

    2000-01-01

    To analyze and compare the heights of first-year school children and their parents, according to ethnic background and socioeconomic status. This is a cross-sectional study of indigenous and non-indigenous school children and their parents, belonging to three levels of social vulnerability: very high (poverty), medium, and very low. An indigenous school child was defined as any child having all four parental surnames of Mapuche origin; non-indigenous were those having Hispanic parental surname. Height was compared using Z scores, using WHO nutritional change reference values. Statistical analysis consisted in comparing differences of mean heights between parents and their children. Differences were assessed using Scheffe's method. Improvement in socioeconomic conditions was associated with increasing mean parental height (p < 0.001), except for indigenous mothers, who showed no height increase. Fathers from highly impoverished counties were 4 cm shorter than those living in very low vulnerability areas; height differences reached 2 cm among mothers (p < 0.001). Indigenous school children showed a positive height gradient with improving socioeconomic conditions (p < 0.001). This was not observed among non-indigenous children. When comparing parental height with children's height, children had a better height/age ratio than their parents (p < 0.01). This was specially evident among indigenous school children, who had on average 1.4 Z scores more than their parents. The upward height gradient related to improved social conditions, and the better height/age ratio seen in children in comparison to their parents, regardless their ethnic background and level of social vulnerability, is encouraging and suggests that interventions directed to the poorest groups, including the indigenous population, must be carried out.

  15. Induction of endothelial cell proliferation by angiogenic factors released by activated monocytes

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

    Pakala, Rajbabu; Watanabe, Takuya; Benedict, Claude R

    2002-06-01

    Introduction: Cell-cell interaction is an essential component of atherosclerotic plaque development. Activated monocytes appear to play a central role in the development of atherosclerosis, not only through foam cell formation but also via the production of various growth factors that induce proliferation of different cell types that are involved in the plaque development. Using serum free co-culture method, we determined the effect of monocytes on endothelial cell proliferation. Methods: Endothelial cell proliferation is determined by the amount of [{sup 3}H]thymidine incorporated in to the DNA. Basic fibroblast growth factor (b-FGF), vascular endothelial growth factor (VEGF) and interleukin-8 (IL-8) levels inmore » the conditioned medium were determined by ELISA. Results: Conditioned medium from unactivated monocytes partially inhibited endothelial cell proliferation, whereas conditioned medium from activated monocytes promoted endothelial cell proliferation. The mitogenic effect of conditioned medium derived from activated monocytes is due to the presence of b-FGF, VEGF and IL-8. Neutralizing antibodies against b-FGF, VEGF and IL-8 partially reversed the mitogenic effect of conditioned medium derived from activated monocytes. When b-FGF, VEGF and IL-8 were immunoprecipitated from conditioned medium derived from activated monocytes, it is less mitogenic to endothelial cells. Conclusion: Activated monocytes may play an important role in the development of atherosclerotic plaque by producing endothelial cell growth factors.« less

  16. wnt3a but not wnt11 supports self-renewal of embryonic stem cells

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

    Singla, Dinender K.; Schneider, David J.; LeWinter, Martin M.

    2006-06-30

    wnt proteins (wnts) promote both differentiation of midbrain dopaminergic cells and self-renewal of haematopoietic stem cells. Mouse embryonic stem (ES) cells can be maintained and self-renew on mouse feeder cell layers or in media containing leukemia inhibitory factor (LIF). However, the effects of wnts on ES cells self-renewal and differentiation are not clearly understood. In the present study, we found that conditioned medium prepared from L cells expressing wnt3a can replace feeder cell layers and medium containing LIF in maintaining ES cells in the proliferation without differentiation (self-renewal) state. By contrast, conditioned medium from NIH3T3 cells expressing wnt11 did not.more » Alkaline phosphatase staining and compact colony formation were used as criteria of cells being in the undifferentiated state. ES cells maintained in medium conditioned by Wnt3a expressing cells underwent freezing and thawing while maintaining properties seen with LIF maintained ES cells. Purified wnt3a did not maintain self-renewal of ES cells for prolonged intervals. Thus, other factors in the medium conditioned by wnt3a expressing cells may have contributed to maintenance of ES cells in a self-renewal state. Pluripotency of ES cells was determined with the use of embryoid bodies in vitro. PD98059, a MEK specific inhibitor, promoted the growth of undifferentiated ES cells maintained in conditioned medium from wnt3a expressing cells. By contrast, the P38 MAPK inhibitor SB230580 did not, suggesting a role for the MEK pathway in self-renewal and differentiation of ES cells maintained in the wnt3a cell conditioned medium. Thus, our results show that conditioned medium from wnt3a but not wnt11 expressing cells can maintain ES cells in self-renewal and in a pluripotent state.« less

  17. Regulating Ultrasound Cavitation in order to Induce Reproducible Sonoporation

    NASA Astrophysics Data System (ADS)

    Mestas, J.-L.; Alberti, L.; El Maalouf, J.; Béra, J.-C.; Gilles, B.

    2010-03-01

    Sonoporation would be linked to cavitation, which generally appears to be a non reproducible and unstationary phenomenon. In order to obtain an acceptable trade-off between cell mortality and transfection, a regulated cavitation generator based on an acoustical cavitation measurement was developed and tested. The medium to be sonicated is placed in a sample tray. This tray is immersed in in degassed water and positioned above the face of a flat ultrasonic transducer (frequency: 445 kHz; intensity range: 0.08-1.09 W/cm2). This technical configuration was admitted to be conducive to standing-wave generation through reflection at the air/medium interface in the well thus enhancing the cavitation phenomenon. Laterally to the transducer, a homemade hydrophone was oriented to receive the acoustical signal from the bubbles. From this spectral signal recorded at intervals of 5 ms, a cavitation index was calculated as the mean of the cavitation spectrum integration in a logarithmic scale, and the excitation power is automatically corrected. The device generates stable and reproducible cavitation level for a wide range of cavitation setpoint from stable cavitation condition up to full-developed inertial cavitation. For the ultrasound intensity range used, the time delay of the response is lower than 200 ms. The cavitation regulation device was evaluated in terms of chemical bubble collapse effect. Hydroxyl radical production was measured on terephthalic acid solutions. In open loop, the results present a great variability whatever the excitation power. On the contrary the closed loop allows a great reproducibility. This device was implemented for study of sonodynamic effect. The regulation provides more reproducible results independent of cell medium and experimental conditions (temperature, pressure). Other applications of this regulated cavitation device concern internalization of different particles (Quantum Dot) molecules (SiRNA) or plasmids (GFP, DsRed) into different types of cells (AT2, RL, LLC…). Preliminary results are presented.

  18. Human umbilical cord tissue-derived mesenchymal stromal cells attenuate remodeling after myocardial infarction by proangiogenic, antiapoptotic, and endogenous cell-activation mechanisms

    PubMed Central

    2014-01-01

    Introduction Among the plethora of cells under investigation to restore a functional myocardium, mesenchymal stromal cells (MSCs) have been granted considerable interest. However, whereas the beneficial effects of bone marrow MSCs (BM-MSCs) in the context of the diseased heart are widely reported, data are still scarce on MSCs from the umbilical cord matrix (UCM-MSCs). Herein we report on the effect of UCM-MSC transplantation to the infarcted murine heart, seconded by the dissection of the molecular mechanisms at play. Methods Human umbilical cord tissue-derived MSCs (UCX®), obtained by using a proprietary technology developed by ECBio, were delivered via intramyocardial injection to C57BL/6 females subjected to permanent ligation of the left descending coronary artery. Moreover, medium produced by cultured UCX® preconditioned under normoxia (CM) or hypoxia (CMH) was collected for subsequent in vitro assays. Results Evaluation of the effects upon intramyocardial transplantation shows that UCX® preserved cardiac function and attenuated cardiac remodeling subsequent to myocardial infarction (MI). UCX® further led to increased capillary density and decreased apoptosis in the injured tissue. In vitro, UCX®-conditioned medium displayed (a) proangiogenic activity by promoting the formation of capillary-like structures by human umbilical vein endothelial cells (HUVECs), and (b) antiapoptotic activity in HL-1 cardiomyocytes subjected to hypoxia. Moreover, in adult murine cardiac Sca-1+ progenitor cells (CPCs), conditioned medium enhanced mitogenic activity while activating a gene program characteristic of cardiomyogenic differentiation. Conclusions UCX® preserve cardiac function after intramyocardial transplantation in a MI murine model. The cardioprotective effects of UCX® were attributed to paracrine mechanisms that appear to enhance angiogenesis, limit the extent of the apoptosis, augment proliferation, and activate a pool of resident CPCs. Overall, these results suggest that UCX® should be considered an alternative cell source when designing new therapeutic approaches to treat MI. PMID:24411922

  19. The anti-apoptotic activity associated with phosphatidylinositol transfer protein alpha activates the MAPK and Akt/PKB pathway.

    PubMed

    Schenning, Martijn; Goedhart, Joachim; Gadella, Theodorus W J; Avram, Diana; Wirtz, Karel W A; Snoek, Gerry T

    2008-10-01

    The conditioned medium (CM) from mouse NIH3T3 fibroblast cells overexpressing phosphatidylinositol transfer protein alpha (PI-TPalpha; SPIalpha cells) demonstrates an increased anti-apoptotic activity compared with CM from wild type NIH3T3 (wtNIH3T3) cells. As previously shown, the anti-apoptotic activity acts by activating a G protein-coupled receptor, most probably a cannabinoid 1 (CB1)-like receptor as the activity was blocked by both pertussis toxin and rimonabant [M. Schenning, C.M. van Tiel, D. Van Manen, J.C. Stam, B.M. Gadella, K.W. Wirtz and G.T. Snoek, Phosphatidylinositol transfer protein alpha regulates growth and apoptosis of NIH3T3 cells: involvement of a cannabinoid 1-like receptor, J. Lipid Res. 45 (2004) 1555-1564]. The CB1 receptor appears to be expressed in mouse fibroblast cells, at levels in the order SPIalpha>wtNIH3T3>SPIbeta cells (i.e. wild type cells overexpressing PI-TPbeta). Upon incubation of SPIbeta cells with the PI-TPalpha-dependent anti-apoptotic factors, both the ERK/MAP kinase and the Akt/PKB pathway are activated in a CB1 receptor dependent manner as shown by Western blotting. In addition, activation of ERK2 was also shown by EYFP-ERK2 translocation to the nucleus, as visualized by confocal laser scanning microscopy. The subsequent activation of the anti-apoptotic transcription factor NF-kappaB is in line with the increased resistance towards UV-induced apoptosis. On the other hand, receptor activation by CM from SPIalpha cells was not linked to phospholipase C activation as the YFP-labelled C2-domain of protein kinase C was not translocated to the plasma membrane of SPIbeta cells as visualized by confocal laser scanning microscopy.

  20. The Interaction between Adult Cardiac Fibroblasts and Embryonic Stem Cell-Derived Cardiomyocytes Leads to Proarrhythmic Changes in In Vitro Cocultures

    PubMed Central

    Trieschmann, Jan; Bettin, Daniel; Haustein, Moritz; Köster, Annette; Molcanyi, Marek; Halbach, Marcel; Hanna, Mira; Fouad, Mariam; Brockmeier, Konrad; Hescheler, Jürgen; Pfannkuche, Kurt; Hannes, Tobias

    2016-01-01

    Transplantation of stem cell-derived cardiomyocytes is one of the most promising therapeutic approaches after myocardial infarction, as loss of cardiomyocytes is virtually irreversible by endogenous repair mechanisms. In myocardial scars, transplanted cardiomyocytes will be in immediate contact with cardiac fibroblasts. While it is well documented how the electrophysiology of neonatal cardiomyocytes is modulated by cardiac fibroblasts of the same developmental stage, it is unknown how adult cardiac fibroblasts (aCFs) affect the function of embryonic stem cell-derived cardiomyocytes (ESC-CMs). To investigate the effects of aCFs on ESC-CM electrophysiology, we performed extra- and intracellular recordings of murine aCF-ESC-CM cocultures. We observed that spontaneous beating behaviour was highly irregular in aCF-ESC-CM cocultures compared to cocultures with mesenchymal stem cells (coefficient of variation of the interspike interval: 40.5 ± 15.2% versus 9.3 ± 2.0%, p = 0.008) and that action potential amplitude and maximal upstroke velocity (V max) were reduced (amplitude: 52.3 ± 1.7 mV versus 65.1 ± 1.5 mV, V max: 7.0 ± 1.0 V/s versus 36.5 ± 5.3 V/s), while action potential duration (APD) was prolonged (APD50: 25.6 ± 1.0 ms versus 16.8 ± 1.9 ms, p < 0.001; APD90: 52.2 ± 1.5 ms versus 43.3 ± 3.3 ms, p < 0.01) compared to controls. Similar changes could be induced by aCF-conditioned medium. We conclude that the presence of aCFs changes automaticity and induces potentially proarrhythmic changes of ESC-CM electrophysiology. PMID:26880949

  1. The Interaction between Adult Cardiac Fibroblasts and Embryonic Stem Cell-Derived Cardiomyocytes Leads to Proarrhythmic Changes in In Vitro Cocultures.

    PubMed

    Trieschmann, Jan; Bettin, Daniel; Haustein, Moritz; Köster, Annette; Molcanyi, Marek; Halbach, Marcel; Hanna, Mira; Fouad, Mariam; Brockmeier, Konrad; Hescheler, Jürgen; Pfannkuche, Kurt; Hannes, Tobias

    2016-01-01

    Transplantation of stem cell-derived cardiomyocytes is one of the most promising therapeutic approaches after myocardial infarction, as loss of cardiomyocytes is virtually irreversible by endogenous repair mechanisms. In myocardial scars, transplanted cardiomyocytes will be in immediate contact with cardiac fibroblasts. While it is well documented how the electrophysiology of neonatal cardiomyocytes is modulated by cardiac fibroblasts of the same developmental stage, it is unknown how adult cardiac fibroblasts (aCFs) affect the function of embryonic stem cell-derived cardiomyocytes (ESC-CMs). To investigate the effects of aCFs on ESC-CM electrophysiology, we performed extra- and intracellular recordings of murine aCF-ESC-CM cocultures. We observed that spontaneous beating behaviour was highly irregular in aCF-ESC-CM cocultures compared to cocultures with mesenchymal stem cells (coefficient of variation of the interspike interval: 40.5 ± 15.2% versus 9.3 ± 2.0%, p = 0.008) and that action potential amplitude and maximal upstroke velocity (V max) were reduced (amplitude: 52.3 ± 1.7 mV versus 65.1 ± 1.5 mV, V max: 7.0 ± 1.0 V/s versus 36.5 ± 5.3 V/s), while action potential duration (APD) was prolonged (APD50: 25.6 ± 1.0 ms versus 16.8 ± 1.9 ms, p < 0.001; APD90: 52.2 ± 1.5 ms versus 43.3 ± 3.3 ms, p < 0.01) compared to controls. Similar changes could be induced by aCF-conditioned medium. We conclude that the presence of aCFs changes automaticity and induces potentially proarrhythmic changes of ESC-CM electrophysiology.

  2. Myostatin is a novel tumoral factor that induces cancer cachexia

    PubMed Central

    Lokireddy, Sudarsanareddy; Wijesoma, Isuru Wijerupage; Bonala, Sabeera; Wei, Meng; Sze, Siu Kwan; McFarlane, Craig; Kambadur, Ravi; Sharma, Mridula

    2012-01-01

    Humoral and tumoral factors collectively promote cancer-induced skeletal muscle wasting by increasing protein degradation. Although several humoral proteins, namely TNFα (tumour necrosis factor α) and IL (interleukin)-6, have been shown to induce skeletal muscle wasting, there is a lack of information regarding the tumoral factors that contribute to the atrophy of muscle during cancer cachexia. Therefore, in the present study, we have characterized the secretome of C26 colon cancer cells to identify the tumoral factors involved in cancer-induced skeletal muscle wasting. In the present study, we show that myostatin, a procachectic TGFβ (transforming growth factor β) superfamily member, is abundantly secreted by C26 cells. Consistent with myostatin signalling during cachexia, treating differentiated C2C12 myotubes with C26 CM (conditioned medium) resulted in myotubular atrophy due to the up-regulation of muscle-specific E3 ligases, atrogin-1 and MuRF1 (muscle RING-finger protein 1), and enhanced activity of the ubiquitin–proteasome pathway. Furthermore, the C26 CM also activated ActRIIB (activin receptor type II B)/Smad and NF-κB (nuclear factor κB) signalling, and reduced the activity of the IGF-I (insulin-like growth factor 1)/PI3K (phosphoinositide 3-kinase)/Akt pathway, three salient molecular features of myostatin action in skeletal muscles. Antagonists to myostatin prevented C26 CM-induced wasting in muscle cell cultures, further confirming that tumoral myostatin may be a key contributor in the pathogenesis of cancer cachexia. Finally, we show that treatment with C26 CM induced the autophagy–lysosome pathway and reduced the number of mitochondria in myotubes. These two previously unreported observations were recapitulated in skeletal muscles collected from C26 tumour-bearing mice. PMID:22621320

  3. Candida krusei isolated from fruit juices ultrafiltration membranes promotes colonization of Escherichia coli O157:H7 and Salmonella enterica on stainless steel surfaces.

    PubMed

    Tarifa, María Clara; Lozano, Jorge Enrique; Brugnoni, Lorena Inés

    2017-02-01

    To clarify the interactions between a common food spoilage yeast and two pathogenic bacteria involved in outbreaks associated with fruit juices, the present paper studies the effect of the interplay of Candida krusei, collected from UF membranes, with Escherichia coli O157:H7 and Salmonella enterica in the overall process of adhesion and colonization of abiotic surfaces. Two different cases were tested: a) co-adhesion by pathogenic bacteria and yeasts, and b) incorporation of bacteria to pre-adhered C. krusei cells. Cultures were made on stainless steel at 25°C using apple juice as culture medium. After 24 h of co-adhesion with C. krusei, both E. coli O157:H7 and S. enterica increased their counts 1.05 and 1.11 log CFU cm 2 , respectively. Similar increases were obtained when incorporating bacteria to pre-adhered cells of Candida. Nevertheless C. krusei counts decreased in both experimental conditions, in a) 0.40 log CFU cm 2 and 0.55 log CFU cm 2 when exposed to E. coli O157:H7 and S. enterica and in b) 0.18 and 0.68 log CFU cm 2 , respectively. This suggests that C. krusei, E. coli O157:H7, and S. enterica have a complex relationship involving physical and chemical interactions on food contact surfaces. This study supports the possibility that pathogen interactions with members of spoilage microbiota, such as C. krusei, might play an important role for the survival and dissemination of E. coli O157:H7 and Salmonella enterica in food-processing environments. Based on the data obtained from the present study, much more attention should be given to prevent the contamination of these pathogens in acidic drinks.

  4. Global trends in significant wave height and marine wind speed from the ERA-20CM

    NASA Astrophysics Data System (ADS)

    Aarnes, Ole Johan; Breivik, Øyvind

    2016-04-01

    The ERA-20CM is one of the latest additions to the ERA-series produced at the European Center for Medium-Range Weather Forecasts (ECMWF). This 10 member ensemble is generated with a version of the Integrated Forecast System (IFS), a coupled atmosphere-wave model. The model integration is run as a AMIP (Atmospheric Model Intercomparison Project) constrained by CMIP5 recommended radiative forcing and different realizations of sea-surface temperature (SST) and sea-ice cover (SIC) prescribed by the HadISST2 (Met Office Hadley Center). While the ERA-20CM is unable to reproduce the actual synoptic conditions, it is designed to offer a realistic statistical representation of the past climate, spanning the period 1899-2010. In this study we investigate global trends in significant wave height and marine wind speed based on ERA-20CM, using monthly mean data, upper percentiles and monthly/annual maxima. The aim of the study is to assess the quality of the trends and how these estimates are affected by different SST and SIC. Global trends are compared against corresponding estimates obtained with ERA-Interim (1979-2009), but also crosschecked against ERA-20C - an ECMWF pilot reanalysis of the 20th-century, known to most trustworthy in the Northern Hemisphere extratropics. Over the period 1900-2009, the 10 member ensemble yields trends mainly within +/- 5% per century. However, significant trends of opposite signs are found locally. Certain areas, like the eastern equatorial Pacific, highly affected by the El Niño Southern Oscillation, show stronger trends. In general, trends based on statistical quantities further into the tail of the distribution are found less reliable.

  5. Effect of spatial variability on solute velocity and dispersion in two soils of the Argentinian Pampas

    NASA Astrophysics Data System (ADS)

    Aparicio, Virginia; Costa, José; Domenech, Marisa; Castro Franco, Mauricio

    2013-04-01

    Predicting how solutes move through the unsaturated zone is essential to determine the potential risk of groundwater contamination (Costa et al., 1994). The estimation of the spatial variability of solute transport parameters, such as velocity and dispersion, enables a more accurate understanding of transport processes. Apparent electrical conductivity (ECa) has been used to characterize the spatial behavior of soil properties. The objective of this study was to characterize the spatial variability of soil transport parameters at field scale using ECa measurements. ECa measurements of 42 ha (Tres Arroyos) and 50 ha (Balcarce) farms were collected for the top 0-30 cm (ECa(s)) soil using the Veris® 3100. ECa maps were generated using geostatistical interpolation techniques. From these maps, three general areas were delineated, named high, medium, and low ECa zones. At each zone, three sub samples were collected. Soil samples were taken at 0-30 cm. Clay content and organic matter (OM) was analyzed. The transport assay was performed in the laboratory using undisturbed soil columns, under controlled conditions of T ° (22 ° C).Br- determinations were performed with a specific Br- electrode. The breakthrough curves were fitted using the model CXTFIT 2.1 (Toride et al., 1999) to estimate the transport parameters Velocity (V) and Dispersion (D). In this study we found no statistical significant differences for V and D between treatments. Also, there were no differences in V and D between sites. The average V and D value was 9.3 cm h-1 and 357.5 cm2 h-2, respectively. Despite finding statistically significant differences between treatments for the other measured physical and chemical properties, in our work it was not possible to detect the spatial variability of solute transport parameters.

  6. [Massive multiplication of coffee (Coffee arabica L. cv. Catimor) through embryogenic cell suspension culture].

    PubMed

    Flermoso-Gallardo, L; Menóndez-Yuffá, A

    2000-01-01

    Cell suspensions offer several advantages as a system for massive propagation because of the high rates of multiplication, the higher homogeneity in the culture conditions and the possibility of automatization. In this study, different experimental conditions were analyzed to establish embryogenic cell suspension cultures of coffee. The best conditions to establish the embryogenic cell suspension cultures of coffee were as follows: coffee leaf sections were cultivated during 12 weeks (Stage I) in a solid medium with the Murashige and Skoog salts, 2 mg/l kinetin and 0.5 mg/l 2,4-dichlorophenoxiacetic acid (medium 1). Under these conditions the explants formed a callus tissue that was transferred to a liquid medium containing 5 mg/l of 6-benzylamlno-purine (medium 2). After 12 days in a shaking liquid medium (Stage II), the cultures were sieved and were maintained In the same media, which was renewed every eight days (Stage III). This method yielded 1884 embryos in 50 ml; placing the embryos under conditions for germination yielded plantlets of normal appearance.

  7. Optimization of an efficient semi-solid culture protocol for sterilization and plant regeneration of Centella asiatica (L.) as a medicinal herb.

    PubMed

    Moghaddam, Sina Siavash; Jaafar, Hawa Binti; Aziz, Maheran Abdul; Ibrahim, Rusli; Rahmat, Asmah Bt; Philip, Elizabeth

    2011-01-01

    The present study investigates the effects of different concentrations, as well as type of plant growth regulators (PGRs) and medium (MS, Duchefa) on the growth and development of Centella asiatica in semi-solid culture. In addition, a protocol for successful sterilization of C.asiatica explants prepared from field-grown plants highly exposed to fungal and bacterial contamination was determined. Results for sterilization treatments revealed that applying HgCl₂ and Plant Preservative Mixture (PPM) with cetrimide, bavistin and trimethoprim which were included after washing with tap water, followed by the addition of PPM in the medium, produced a very satisfactory result (clean culture 90 ± 1.33%) and TS5 (decon + cetrimide 1% + bavistin 150 mg/L + trimethoprim 50 mg/L + HgCl₂0.1% + PPM 2% soak and 2 mL/L in medium) was hence chosen as the best method of sterilization for C.asiatica. The synergistic combination of 6 benzylaminopurine (BAP) and 1-naphthaleneacetic acid (NAA) in concentrations of 2 mg/L and 0.1 mg/L, respectively, in Duchefa medium compared with MS induced the most optimal percentage of sprouted shoots (93 ± 0.667), number of shoots (5.2 ± 0.079) and nodes (4 ± 0.067) per explant, leaf per explant (14 ± 0.107) and shoot length (4.1 ± 0.67 cm). Furthermore, optimum rooting frequency (95.2 ± 0.81%), the number of roots/shoot (7.5 ± 0.107) and the mean root length (4.5 ± 0.133 cm) occurred for shoots that were cultured on full-strength MS medium containing 0.5 mg/L indole-3-butyric acid (IBA). In this study, the acclimatized plantlets were successfully established with almost 85% survival. The findings of this study have proven an efficient medium and PGR concentration for the mass propagation of C.asiatica. These findings would be useful in micropropagation and ex situ conservation of this plant.

  8. Trophic factors from adipose tissue-derived multi-lineage progenitor cells promote cytodifferentiation of periodontal ligament cells

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

    Sawada, Keigo; Takedachi, Masahide, E-mail: takedati@dent.osaka-u.ac.jp; Yamamoto, Satomi

    Stem and progenitor cells are currently being investigated for their applicability in cell-based therapy for periodontal tissue regeneration. We recently demonstrated that the transplantation of adipose tissue-derived multi-lineage progenitor cells (ADMPCs) enhances periodontal tissue regeneration in beagle dogs. However, the molecular mechanisms by which transplanted ADMPCs induce periodontal tissue regeneration remain to be elucidated. In this study, trophic factors released by ADMPCs were examined for their paracrine effects on human periodontal ligament cell (HPDL) function. ADMPC conditioned medium (ADMPC-CM) up-regulated osteoblastic gene expression, alkaline phosphatase activity and calcified nodule formation in HPDLs, but did not significantly affect their proliferative response.more » ADMPCs secreted a number of growth factors, including insulin-like growth factor binding protein 6 (IGFBP6), hepatocyte growth factor and vascular endothelial growth factor. Among these, IGFBP6 was most highly expressed. Interestingly, the positive effects of ADMPC-CM on HPDL differentiation were significantly suppressed by transfecting ADMPCs with IGFBP6 siRNA. Our results suggest that ADMPCs transplanted into a defect in periodontal tissue release trophic factors that can stimulate the differentiation of HPDLs to mineralized tissue-forming cells, such as osteoblasts and cementoblasts. IGFBP6 may play crucial roles in ADMPC-induced periodontal regeneration. - Highlights: • ADMPC-derived humoral factors stimulate cytodifferentiation of HPDLs. • ADMPCs secret growth factors including IGFBP6, VEGF and HGF. • IGFBP6 is involved in the promotion effect of ADMPC-CM on HPDL cytodifferentiation.« less

  9. Light based technologies for microbial inactivation of liquids, bead surfaces and powdered infant formula.

    PubMed

    Arroyo, Cristina; Dorozko, Anna; Gaston, Edurne; O'Sullivan, Michael; Whyte, Paul; Lyng, James G

    2017-10-01

    This study evaluates the potential of continuous wave Ultraviolet C light (UV-C) and broad-spectrum intense pulsed light (in this study referred to as High Intensity Light Pulses, HILP) for the inactivation of pathogens of public concern in powdered infant formula (PIF) producers. To achieve this goal a sequential set of experiments were performed, firstly in clear liquid media, secondly on the surface of spherical beads under agitation and, finally in PIF. L. innocua was the most sensitive microorganism to both technologies under all conditions studied with reductions exceeding 4 log 10 cycles in PIF. In the clear liquid medium, the maximum tolerance to light was observed for C. sakazakii against UV-C light and for B. subtilis spores against HILP, with a fluence of approximately 17 mJ/cm 2 required for a 1 log 10 cycle inactivation (D value) of each species. In PIF it was possible to inactivate >99% of the vegetative cell populations by HILP with a fluence of 199 mJ/cm 2 and of B. subtilis spores by doubling the fluence. By contrast, for UV-C treatments a fluence of 2853 mJ/cm 2 was needed for 99.9% reduction of C. sakazakii, which was the most light-resistant microorganism to UV-C. Results here obtained clearly show the potential for light-based interventions to improve PIF microbiological safety. Copyright © 2017 Elsevier Ltd. All rights reserved.

  10. Increased apoptosis and peripheral blood mononuclear cell suppression of bone marrow mesenchymal stem cells in severe aplastic anemia.

    PubMed

    Chao, Yu-Hua; Lin, Chiao-Wen; Pan, Hui-Hsien; Yang, Shun-Fa; Weng, Te-Fu; Peng, Ching-Tien; Wu, Kang-Hsi

    2018-06-05

    Although immune-mediated pathogenesis is considered an important aspect of severe aplastic anemia (SAA), its underlying mechanisms remain unclear. Mesenchymal stem cells (MSCs) are essential to the formation of specialized microenvironments in the bone marrow (BM), and MSC insufficiency can trigger the development of SAA. To find MSC alterations in the SAA BM, we compared BM MSCs from five children with SAA and five controls. Peripheral blood mononuclear cells (PBMCs) were cocultured with MSCs to evaluate the supportive effects of MSCs on hematopoiesis. Cytometric bead array immunoassay was used to determine cytokine excretion by MSCs. The immune functions of MSCs and their conditioned medium (CM) were evaluated by PBMC proliferation assays. SAA MSCs were characterized by a high percentage of cells in the abnormal sub-G1 phase of the cell cycle, which suggests an increased rate of apoptosis in SAA MSCs. In comparison with control MSCs, PBMCs cocultured with SAA MSCs displayed significantly reduced PBMC proliferation (P = 0.009). Aberrant cytokine profiles were secreted by SAA MSCs, with increased concentrations of interleukin-6, interferon-γ, tumor necrosis factor-α, and interleukin-1β in the CM. PBMC proliferation assays demonstrated additional immunosuppressive effects of SAA MSCs (P = 0.016) and their CM (P = 0.013). Our data revealed increased apoptosis and PBMC suppression of SAA MSCs. The alterations of MSCs may contribute to the formation of functionally abnormal microenvironments in SAA BM. © 2018 Wiley Periodicals, Inc.

  11. Microenvironment mesenchymal cells protect ovarian cancer cell lines from apoptosis by inhibiting XIAP inactivation

    PubMed Central

    Castells, M; Milhas, D; Gandy, C; Thibault, B; Rafii, A; Delord, J-P; Couderc, B

    2013-01-01

    Epithelial ovarian carcinoma is characterized by high frequency of recurrence (70% of patients) and carboplatin resistance acquisition. Carcinoma-associated mesenchymal stem cells (CA-MSC) have been shown to induce ovarian cancer chemoresistance through trogocytosis. Here we examined CA-MSC properties to protect ovarian cancer cells from carboplatin-induced apoptosis. Apoptosis was determined by Propidium Iodide and Annexin-V-FITC labelling and poly-ADP-ribose polymerase cleavage analysis. We showed a significant increase of inhibitory concentration 50 and a 30% decrease of carboplatin-induced apoptosis in ovarian cancer cells incubated in the presence of CA-MSC-conditioned medium (CM). A molecular analysis of apoptosis signalling pathway in response to carboplatin revealed that the presence of CA-MSC CM induced a 30% decrease of effector caspases-3 and -7 activation and proteolysis activity. CA-MSC secretions promoted Akt and X-linked inhibitor of apoptosis protein (XIAP; caspase inhibitor from inhibitor of apoptosis protein (IAP) family) phosphorylation. XIAP depletion by siRNA strategy permitted to restore apoptosis in ovarian cancer cells stimulated by CA-MSC CM. The factors secreted by CA-MSC are able to confer chemoresistance to carboplatin in ovarian cancer cells through the inhibition of effector caspases activation and apoptosis blockade. Activation of the phosphatidylinositol 3-kinase (PI3K)/Akt signalling pathway and the phosphorylation of its downstream target XIAP underlined the implication of this signalling pathway in ovarian cancer chemoresistance. This study reveals the potentialities of targeting XIAP in ovarian cancer therapy. PMID:24176845

  12. Enhancement of the thermal transport in a culture medium with Au nanoparticles

    NASA Astrophysics Data System (ADS)

    Jiménez-Pérez, J. L.; Fuentes, R. Gutierrez; Alvarado, E. Maldonado; Ramón-Gallegos, E.; Cruz-Orea, A.; Tánori-Cordova, J.; Mendoza-Alvarez, J. G.

    2008-11-01

    In this work, it is reported the gold nanoparticles synthesis, their characterization, and their application to the enhancement of the thermal transport in a cellular culture medium. The Au nanoparticles (NPs), with average size of 10 nm, contained into a culture medium (DMEM (1)/F12(1)) (CM) increased considerably the heat transfer in the medium. Thermal lens spectrometry (TLS) was used to measure the thermal diffusivity of the nanofluids. The characteristic time constant of the transient thermal lens was obtained by fitting the theoretical expression, for transient thermal lens, to the experimental data. Our results show that the thermal diffusivity of the culture medium is highly sensitive to the Au nanoparticle concentration and size. The ability to modify the thermal properties to nanometer scale becomes very important in medical applications as in the case of cancer treatment by using photodynamic therapy (PDT). A complementary study with UV-vis and TEM techniques was performed to characterize the Au nanoparticles.

  13. An investigation into the stability of commercial versus MG63-derived hepatocyte growth factor under flow cultivation conditions.

    PubMed

    Meneghello, Giulia; Storm, Michael P; Chaudhuri, Julian B; De Bank, Paul A; Ellis, Marianne J

    2015-03-01

    The scale-up of tissue engineering cell culture must ensure that conditions are maintained while also being cost effective. Here we analyse the stability of hepatocyte growth factor (HGF) to investigate whether concentrations change under dynamic conditions, and compare commercial recombinant human HGF as an additive in 'standard medium', to HGF secreted by the osteosarcoma cell line MG63 as a 'preconditioned medium'. After 3 h under flow conditions, HGF in the standard medium degraded to 40% of its original concentration but HGF in the preconditioned medium remained at 100%. The concentration of secreted HGF was 10 times greater than the working concentration of commercially-available HGF. Thus HGF within this medium has increased stability; MG63-derived HGF should therefore be investigated as a cost-effective alternative to current lyophilised powders for use in in vitro models. Furthermore, we recommend that those intending to use HGF (or other growth factors) should consider similar stability testing before embarking on experiments with media flow.

  14. Model of Heat Exchangers for Waste Heat Recovery from Diesel Engine Exhaust for Thermoelectric Power Generation

    NASA Astrophysics Data System (ADS)

    Baker, Chad; Vuppuluri, Prem; Shi, Li; Hall, Matthew

    2012-06-01

    The performance and operating characteristics of a hypothetical thermoelectric generator system designed to extract waste heat from the exhaust of a medium-duty turbocharged diesel engine were modeled. The finite-difference model consisted of two integrated submodels: a heat exchanger model and a thermoelectric device model. The heat exchanger model specified a rectangular cross-sectional geometry with liquid coolant on the cold side, and accounted for the difference between the heat transfer rate from the exhaust and that to the coolant. With the spatial variation of the thermoelectric properties accounted for, the thermoelectric device model calculated the hot-side and cold-side heat flux for the temperature boundary conditions given for the thermoelectric elements, iterating until temperature and heat flux boundary conditions satisfied the convection conditions for both exhaust and coolant, and heat transfer in the thermoelectric device. A downhill simplex method was used to optimize the parameters that affected the electrical power output, including the thermoelectric leg height, thermoelectric n-type to p-type leg area ratio, thermoelectric leg area to void area ratio, load electrical resistance, exhaust duct height, coolant duct height, fin spacing in the exhaust duct, location in the engine exhaust system, and number of flow paths within the constrained package volume. The calculation results showed that the configuration with 32 straight fins was optimal across the 30-cm-wide duct for the case of a single duct with total height of 5.5 cm. In addition, three counterflow parallel ducts or flow paths were found to be an optimum number for the given size constraint of 5.5 cm total height, and parallel ducts with counterflow were a better configuration than serpentine flow. Based on the reported thermoelectric properties of MnSi1.75 and Mg2Si0.5Sn0.5, the maximum net electrical power achieved for the three parallel flow paths in a counterflow arrangement was 1.06 kW for package volume of 16.5 L and exhaust flow enthalpy flux of 122 kW.

  15. Human spermatogonial stem cells display limited proliferation in vitro under mouse spermatogonial stem cell culture conditions.

    PubMed

    Medrano, Jose V; Rombaut, Charlotte; Simon, Carlos; Pellicer, Antonio; Goossens, Ellen

    2016-11-01

    To study the ability of human spermatogonial stem cells (hSSCs) to proliferate in vitro under mouse spermatogonial stem cell (mSSC) culture conditions. Experimental basic science study. Reproductive biology laboratory. Cryopreserved testicular tissue with normal spermatogenesis obtained from three donors subjected to orchiectomy due to a prostate cancer treatment. Testicular cells used to create in vitro cell cultures corresponding to the following groups: [1] unsorted human testicular cells, [2] differentially plated human testicular cells, and [3] cells enriched with major histocompatibility complex class 1 (HLA - )/epithelial cell surface antigen (EPCAM + ) in coculture with inactivated testicular feeders from the same patient. Analyses and characterization including immunocytochemistry and quantitative reverse-transcription polymerase chain reaction for somatic and germ cell markers, testosterone and inhibin B quantification, and TUNEL assay. Putative hSSCs appeared in singlets, doublets, or small groups of up to four cells in vitro only when testicular cells were cultured in StemPro-34 medium supplemented with glial cell line-derived neurotrophic factor (GDNF), leukemia inhibitory factor (LIF), basic fibroblast growth factor (bFGF), and epidermal growth factor (EGF). Fluorescence-activated cell sorting with HLA - /EPCAM + resulted in an enrichment of 27% VASA + /UTF1 + hSSCs, compared to 13% in unsorted controls. Coculture of sorted cells with inactivated testicular feeders gave rise to an average density of 112 hSSCs/cm 2 after 2 weeks in vitro compared with unsorted cells (61 hSSCs/cm 2 ) and differentially plated cells (49 hSSCS/cm 2 ). However, putative hSSCs rarely stained positive for the proliferation marker Ki67, and their presence was reduced to the point of almost disappearing after 4 weeks in vitro. We found that hSSCs show limited proliferation in vitro under mSSC culture conditions. Coculture of HLA - /EPCAM + sorted cells with testicular feeders improved the germ cell/somatic cell ratio. Copyright © 2016 American Society for Reproductive Medicine. Published by Elsevier Inc. All rights reserved.

  16. In vitro efficacy and release study with anti-inflammatory drugs incorporated in adhesive transdermal drug delivery systems.

    PubMed

    Meyer, Stefanie; Peters, Nils; Mann, Tobias; Wolber, Rainer; Pörtner, Ralf; Nierle, Jens

    2014-04-01

    The topical application of two different anti-inflammatory extracts incorporated in adhesive transdermal drug delivery systems (TDDSs) was investigated. Therefore, anti-inflammatory properties and percutaneous absorption behavior of adhesive TDDSs were characterized in vitro conducting experiments with a dermatologically relevant human skin model. Anti-inflammatory efficacy against UV irradiation of both TDDSs was determined in vitro with EpiDerm™. The reduction of the release of proinflammatory cytokines by topically applied TDDSs was compared with the reduction during the presence of the specific cyclooxygenase inhibitor diclofenac in the culture medium. A similar anti-inflammatory efficacy of the topically applied TDDSs in comparison with the use of diclofenac in the culture medium should be achieved. Furthermore, percutaneous absorption in efficacy tests was compared with percutaneous absorption in diffusion studies with porcine cadaver skin. Both the topically applied TDDSs showed a significant anti-inflammatory activity. Permeation coefficients through the stratum corneum and the epidermis gained from the release studies on porcine cadaver skin (Magnolia: 2.23·10(-5) cm/h, licorice: 4.68·10(-6) cm/h) were approximately five times lower than the permeation coefficients obtained with the EpiDerm™ skin model (Magnolia: 9.48·10(-5) cm/h, licorice: 24.0·10(-6) cm/h). Therefore, an adjustment of drug doses during experiments with the EpiDerm™ skin model because of weaker skin barrier properties should be considered.

  17. SU-F-I-13: Correction Factor Computations for the NIST Ritz Free Air Chamber for Medium-Energy X Rays

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

    Bergstrom, P

    Purpose: The National Institute of Standards and Technology (NIST) uses 3 free-air chambers to establish primary standards for radiation dosimetry at x-ray energies. For medium-energy × rays, the Ritz free-air chamber is the main measurement device. In order to convert the charge or current collected by the chamber to the radiation quantities air kerma or air kerma rate, a number of correction factors specific to the chamber must be applied. Methods: We used the Monte Carlo codes EGSnrc and PENELOPE. Results: Among these correction factors are the diaphragm correction (which accounts for interactions of photons from the x-ray source inmore » the beam-defining diaphragm of the chamber), the scatter correction (which accounts for the effects of photons scattered out of the primary beam), the electron-loss correction (which accounts for electrons that only partially expend their energy in the collection region), the fluorescence correction (which accounts for ionization due to reabsorption ffluorescence photons and the bremsstrahlung correction (which accounts for the reabsorption of bremsstrahlung photons). We have computed monoenergetic corrections for the NIST Ritz chamber for the 1 cm, 3 cm and 7 cm collection plates. Conclusion: We find good agreement with other’s results for the 7 cm plate. The data used to obtain these correction factors will be used to establish air kerma and it’s uncertainty in the standard NIST x-ray beams.« less

  18. Transformation of proanthocyanidin A2 to its isomers under different physiological pH conditions and common cell culture medium.

    PubMed

    Lu, Wen-Chien; Huang, Wei-Ting; Kumaran, Alaganandam; Ho, Chi-Tang; Hwang, Lucy Sun

    2011-06-08

    Proanthocyanidins constitute an important class of polyphenols ubiquitously found in plants. They have been extensively studied for their antioxidant capacity and bioactivity in vitro and in animal models. However, their stability under different pH conditions and in cell culture medium has not been well documented. In the present study, it was observed that proanthocyanidin A2 (PA2) was relatively more stable in acidic condition than in weak alkaline condition. PA2 was also quite unstable in basal-Dulbecco's Modified Eagle medium (b-DMEM medium) at 37 °C. The addition of PA2 to the cell culture medium accelerated its epimerization with a half-life of <15 min, and ethylenediaminetetraacetic acid (EDTA) could not stop the reaction. The results also demonstrated that the major isomers transformed in the weak alkaline condition or cell culture medium at 37 °C were identified as epicatechin-(4β→8; 2β→O→7)-ent-catechin (proanthocyanidin A4) and epicatechin-(4β→6; 2β→O→7)-ent-catechin. The rates of transformation were dependent on the pH or the components of the medium. Therefore, the results obtained for PA2 in the cell culture bioassays, which were usually carried out for 24 h, might not represent the true activity of the original PA2. The stability and transformation of PA2 should be considered when the bioactivity of PA2 is evaluated in a given cell culture system.

  19. The astrophysical consequences of intervening galaxy gas on fast radio bursts

    NASA Astrophysics Data System (ADS)

    Prochaska, J. Xavier; Neeleman, Marcel

    2018-02-01

    We adopt and analyze results on the incidence and physical properties of damped Ly$\\alpha$ systems (DLAs) to predict the astrophysical impact of gas in galaxies on observations of Fast Radio Bursts (FRBs). Three DLA measures form the basis of this analysis: (i) the HI column density distribution, parameterized as a double power-law; (ii) the incidence of DLAs with redshift (derived here), $\\ell(z)=A+B \\arctan(z-C)$ with $A=0.236_{-0.021}^{+0.016}, B=0.168_{-0.017}^{+0.010}, C=2.87_{-0.13}^{+0.17}$ and (iii) the electron density, parameterized as a log-normal deviate with mean $10^{-2.6} cm^{-3}$ and dispersion 0.3dex. Synthesizing these results, we estimate that the average rest-frame dispersion measure from the neutral medium of a single, intersecting galaxy is DM$^{NM}_{DLA}=0.25$ pc/cm^3. Analysis of AlIII and CII* absorption limits the putative warm ionized medium to contribute DM$^{WIM}_{DLA}<20$pc/cm^3. Given the low incidence of DLAs, we find that a population of FRBs at z=2 will incur DM(z=2)=0.01 pc/cm^3 on average, with a 99% c.l. upper bound of 0.22 pc/cm^3. Assuming that turbulence of the ISM in external galaxies is qualitatively similar to our Galaxy, we estimate that the angular broadening of an FRB by intersecting galaxies is negligible ($\\theta<0.1$mas). The temporal broadening is also predicted to be small, $\\tau \\approx 0.3$ms for a z=1 galaxy intersecting a z=2 FRB for an observing frequency of $\

  20. Equatorial secondary cosmic ray observatory to study space weather and terrestrial events

    NASA Astrophysics Data System (ADS)

    Vichare, Geeta; Bhaskar, Ankush; Datar, Gauri; Raghav, Anil; Nair, K. U.; Selvaraj, C.; Ananthi, M.; Sinha, A. K.; Paranjape, M.; Gawade, T.; Anil Kumar, C. P.; Panneerselvam, C.; Sathishkumar, S.; Gurubaran, S.

    2018-05-01

    Recently, equatorial secondary cosmic ray observatory has been established at Equatorial Geophysical Research Laboratory (EGRL), Tirunelveli, (Geographic Coordinates: 8.71°N, 77.76°E), to study secondary cosmic rays (SCR) produced due to the interaction of primary cosmic rays with the Earth's atmosphere. EGRL is a regional center of Indian Institute of Geomagnetism (IIG), located near the equator in the Southern part of India. Two NaI(Tl) scintillation detectors are installed inside the temperature controlled environment. One detector is cylindrical in shape of size 7.62 cm × 7.62 cm and another one is rectangular cuboid of 10.16 cm × 10.16 cm × 40.64 cm size. Besides NaI(Tl) detectors, various other research facilities such as the Geomagnetic observatory, Medium Frequency Radar System, Digital Ionosonde, All-sky airglow imager, Atmospheric electricity laboratory to measure the near-Earth atmospheric electric fields are also available at EGRL. With the accessibility of multi- instrument facilities, the objective is set to understand the relationship between SCR and various atmospheric and ionospheric processes, during space weather and terrestrial events. For gamma-ray spectroscopy, it is important to test the performance of the NaI(Tl) scintillation detectors and to calibrate the gamma-ray spectrum in terms of energy. The present article describes the details of the experimental setup installed near the equator to study cosmic rays, along with the performance testing and calibration of the detectors under various conditions. A systematic shift in the gain is observed with varying temperature of the detector system. It is found that the detector's response to the variations in the temperature is not just linear or non-linear type, but it depends on the history of the variation, indicating temperature hysteresis effects on NaI detector and PMT system. This signifies the importance of isothermal environment while studying SCR flux using NaI(Tl) detectors, especially for the experiments conducted during daytime such as solar eclipses etc.

  1. Effect of culture medium, host strain and oxygen transfer on recombinant Fab antibody fragment yield and leakage to medium in shaken E. coli cultures.

    PubMed

    Ukkonen, Kaisa; Veijola, Johanna; Vasala, Antti; Neubauer, Peter

    2013-07-29

    Fab antibody fragments in E. coli are usually directed to the oxidizing periplasmic space for correct folding. From periplasm Fab fragments may further leak into extracellular medium. Information on the cultivation parameters affecting this leakage is scarce, and the unpredictable nature of Fab leakage is problematic regarding consistent product recovery. To elucidate the effects of cultivation conditions, we investigated Fab expression and accumulation into either periplasm or medium in E. coli K-12 and E. coli BL21 when grown in different types of media and under different aeration conditions. Small-scale Fab expression demonstrated significant differences in yield and ratio of periplasmic to extracellular Fab between different culture media and host strains. Expression in a medium with fed-batch-like glucose feeding provided highest total and extracellular yields in both strains. Unexpectedly, cultivation in baffled shake flasks at 150 rpm shaking speed resulted in higher yield and accumulation of Fabs into culture medium as compared to cultivation at 250 rpm. In the fed-batch medium, extracellular fraction in E. coli K-12 increased from 2-17% of total Fab at 250 rpm up to 75% at 150 rpm. This was partly due to increased lysis, but also leakage from intact cells increased at the lower shaking speed. Total Fab yield in E. coli BL21 in glycerol-based autoinduction medium was 5 to 9-fold higher at the lower shaking speed, and the extracellular fraction increased from ≤ 10% to 20-90%. The effect of aeration on Fab localization was reproduced in multiwell plate by variation of culture volume. Yield and leakage of Fab fragments are dependent on expression strain, culture medium, aeration rate, and the combination of these parameters. Maximum productivity in fed-batch-like conditions and in autoinduction medium is achieved under sufficiently oxygen-limited conditions, and lower aeration also promotes increased Fab accumulation into extracellular medium. These findings have practical implications for screening applications and small-scale Fab production, and highlight the importance of maintaining consistent aeration conditions during scale-up to avoid changes in product yield and localization. On the other hand, the dependency of Fab leakage on cultivation conditions provides a practical way to manipulate Fab localization.

  2. Effect of culture medium, host strain and oxygen transfer on recombinant Fab antibody fragment yield and leakage to medium in shaken E. coli cultures

    PubMed Central

    2013-01-01

    Background Fab antibody fragments in E. coli are usually directed to the oxidizing periplasmic space for correct folding. From periplasm Fab fragments may further leak into extracellular medium. Information on the cultivation parameters affecting this leakage is scarce, and the unpredictable nature of Fab leakage is problematic regarding consistent product recovery. To elucidate the effects of cultivation conditions, we investigated Fab expression and accumulation into either periplasm or medium in E. coli K-12 and E. coli BL21 when grown in different types of media and under different aeration conditions. Results Small-scale Fab expression demonstrated significant differences in yield and ratio of periplasmic to extracellular Fab between different culture media and host strains. Expression in a medium with fed-batch-like glucose feeding provided highest total and extracellular yields in both strains. Unexpectedly, cultivation in baffled shake flasks at 150 rpm shaking speed resulted in higher yield and accumulation of Fabs into culture medium as compared to cultivation at 250 rpm. In the fed-batch medium, extracellular fraction in E. coli K-12 increased from 2-17% of total Fab at 250 rpm up to 75% at 150 rpm. This was partly due to increased lysis, but also leakage from intact cells increased at the lower shaking speed. Total Fab yield in E. coli BL21 in glycerol-based autoinduction medium was 5 to 9-fold higher at the lower shaking speed, and the extracellular fraction increased from ≤ 10% to 20-90%. The effect of aeration on Fab localization was reproduced in multiwell plate by variation of culture volume. Conclusions Yield and leakage of Fab fragments are dependent on expression strain, culture medium, aeration rate, and the combination of these parameters. Maximum productivity in fed-batch-like conditions and in autoinduction medium is achieved under sufficiently oxygen-limited conditions, and lower aeration also promotes increased Fab accumulation into extracellular medium. These findings have practical implications for screening applications and small-scale Fab production, and highlight the importance of maintaining consistent aeration conditions during scale-up to avoid changes in product yield and localization. On the other hand, the dependency of Fab leakage on cultivation conditions provides a practical way to manipulate Fab localization. PMID:23895637

  3. Thermal behavior modeling of a cabinet direct solar dryer as influenced by sensible heat storage in a fractured porous medium

    NASA Astrophysics Data System (ADS)

    Sandali, Messaoud; Boubekri, Abdelghani; Mennouche, Djamel

    2018-05-01

    Numerical simulation method has been employed to improve the thermal performance of cabinet direct solar dryer. The present study focused on the numerical simulation of a direct solar dryer with integration of a flat layer of fractured porous medium above the absorber plate in the aim to store thermal energy by sensible heat. Several calculations were conducted, using the finite volume method with a two-dimensional unsteady model implemented in the Fluent CFD software. The porous medium has been integrated with different thickness to show the influence of the medium thickness on the thermal performance of solar dryer. Different kinds of materials have been tested and studied. The effect of porosity of porous medium has been studied. The obtained results showed that the temperature of drying air is increased by 4°C with integration of porous medium. The increasing in the thickness of the porous medium by 1cm leads to increase the temperature of drying air by 2°C. The increasing of the medium porosity by 10% leads to decrease the temperature of drying air by 1°C. The best material is the one that has a highest specific heat and thermal conductivity.

  4. Simple Cadmium Sulfide Compound with Stable 95 % Selectivity for Carbon Dioxide Electroreduction in Aqueous Medium.

    PubMed

    Li, Yu Hang; Cheng, Ling; Liu, Peng Fei; Zhang, Le; Zu, Meng Yang; Wang, Chong Wu; Jin, Yan Huan; Cao, Xiao Ming; Yang, Hua Gui; Li, Chunzhong

    2018-05-09

    A simple cadmium sulfide nanomaterial is found to be an efficient and stable electrocatalyst for CO 2 reduction in aqueous medium for more than 40 h with a steady CO faradaic efficiency of approximately 95 %. Moreover, it can realize a current density of -10 mA cm -2 at an overpotential of -0.55 V on a porous substrate with similar selectivity. Theoretical and experimental results confirm that the high selectivity for CO 2 reduction is due to its (0 0 0 2) face with sulfur vacancies that prefers CO 2 molecule reduction in aqueous medium. © 2018 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.

  5. Control of wave propagation in a biological excitable medium by an external electric field.

    PubMed

    Sebestikova, Lenka; Slamova, Elena; Sevcikova, Hana

    2005-03-01

    We present an experimental evidence of effects of external electric fields (EFs) on the velocity of pulse waves propagating in a biological excitable medium. The excitable medium used is formed by a layer of starving cells of Dictyostelium discoideum through which the waves of increased concentration of cAMP propagate by reaction-diffusion mechanism. External dc EFs of low intensities (up to 5 V/cm) are shown to speed up the propagation of cAMP waves towards the positive electrode and slow it down towards the negative electrode. Electric fields were also found to support an emergence of new centers, emitting cAMP waves, in front of cAMP waves propagating towards the negative electrode.

  6. Development of thermally controlled HALNA DPSSL for inertial fusion energy

    NASA Astrophysics Data System (ADS)

    Matsumoto, Osamu; Yasuhara, Ryo; Kurita, Takashi; Ikegawa, Tadashi; Sekine, Takashi; Kawashima, Toshiyuki; Kawanaka, Junji; Norimatsu, Takayoshi; Miyanaga, Noriaki; Izawa, Yasukazu; Nakatsuka, Masahiro; Miyamoto, Masahiro; Kan, Hirofumi; Furukawa, Hiroyuki; Motokoshi, Shinji

    2006-02-01

    We have been developing a high average-power laser system for science and industry applications that can generate an output of 20 J per pulse at 10-Hz operation. Water-cooled Nd:glass zig-zag slab is pumped with 803-nm AlGaAs laser-diode modules. To efficiently extract energy from the laser medium, the laser beam alternately passes through dual zig-zag slab amplifier modules. Twin LD modules equipped on each slab amplifier module pump the laser medium with a peak power density of 2.5 kW/cm2. In high power laser system, thermal load in the laser medium causes serious thermal effects. We arranged cladding glasses on the top and bottom of the laser slab to reduce thermal effects.

  7. Fusion pumped laser

    DOEpatents

    Pappas, Daniel S.

    1989-01-01

    Apparatus is provided for generating energy in the form of laser radiation. A tokamak fusion reactor is provided for generating a long, or continuous, pulse of high-energy neutrons. The tokamak design provides a temperature and a magnetic field which is effective to generate a neutron flux of at least 10.sup.15 neutrons/cm.sup.2.s. A conversion medium receives neutrons from the tokamak and converts the high-energy neutrons to an energy source with an intensity and an energy effective to excite a preselected lasing medium. The energy source typically comprises fission fragments, alpha particles, and radiation from a fission event. A lasing medium is provided which is responsive to the energy source to generate a population inversion which is effective to support laser oscillations for generating output radiation.

  8. Aggressive and foraging behavioral interactions among ruffe

    USGS Publications Warehouse

    Savino, Jacqueline F.; Kostich, Melissa J.

    2000-01-01

    The ruffe, Gymnocephalus cernuus, is a nonindigenous percid in the Great Lakes. Ruffe are aggressive benthivores and forage over soft substrates. Laboratory studies in pools (100 cm in diameter, 15 cm water depth) were conducted to determine whether fish density (low = 2, medium = 4, high = 6 ruffe per pool) changed foraging and aggressive behaviors with a limited food supply of chironomid larvae. All fish densities demonstrated a hierarchy based on aggressive interactions, but ruffe were most aggressive at low and high fish densities. Time spent in foraging was lowest at the low fish density. The best forager at the low fish density was the most aggressive individual, but the second most aggressive fish at the medium and high fish density was the best forager and also the one chased most frequently. A medium fish density offered the best energetic benefits to ruffe by providing the lowest ratio of time spent in aggression to that spent foraging. Based on our results, ruffe should grow best at an intermediate density. With high ruffe densities, we would also expect disparity in size as the more aggressive fish are able to garner a disproportionate amount of the resources. Alternatively, as the Great Lakes are a fairly open system, ruffe could migrate out of one area to colonize another as populations exceed optimal densities.

  9. Micropropagation of Hedychium coronarium J. Koenig through rhizome bud.

    PubMed

    Mohanty, Pritam; Behera, Shashikanta; Swain, Swasti S; Barik, Durga P; Naik, Soumendra K

    2013-10-01

    An optimized protocol was developed for in vitro plant regeneration of a medicinally important herb Hedychium coronarium J. Koenig using sprouted buds of rhizomes. The rhizomes with sprouted bud were inoculated on Murashige and Skoog (Physiol Plant 15:473-497, 1962) medium (MS) supplemented with either N(6)-benzyladenine (BA) alone (1.0-4.0 mg L(-1)) or in combination with 0.5 mg L(-1) naphthalene acetic acid (NAA). Of these combinations, MS supplemented with a combination of 2.0 mg L(-1) BA and 0.5 mg L(-1) NAA was most effective. In this medium, best shoots (3.6) and roots (4.0) regeneration was observed simultaneously with an average shoot and root length of 4.7 cm and 4.2 cm respectively. Regeneration of shoots and roots in the same medium at the same time (One step shoot and root regeneration) reduced the time for production of in vitro plantlets and eliminates the media cost of rooting. Cent-percent (100 %) success in plant establishment was observed in both gradual acclimatization process as well as when plants were directly transferred to outdoor in clay pots containing a mixture of garden soil and sand (2:1) without any sequential acclimatization stage.

  10. Integrated Microfluidic Flow-Through Microbial Fuel Cells

    PubMed Central

    Jiang, Huawei; Ali, Md. Azahar; Xu, Zhen; Halverson, Larry J.; Dong, Liang

    2017-01-01

    This paper reports on a miniaturized microbial fuel cell with a microfluidic flow-through configuration: a porous anolyte chamber is formed by filling a microfluidic chamber with three-dimensional graphene foam as anode, allowing nutritional medium to flow through the chamber to intimately interact with the colonized microbes on the scaffolds of the anode. No nutritional media flow over the anode. This allows sustaining high levels of nutrient utilization, minimizing consumption of nutritional substrates, and reducing response time of electricity generation owing to fast mass transport through pressure-driven flow and rapid diffusion of nutrients within the anode. The device provides a volume power density of 745 μW/cm3 and a surface power density of 89.4 μW/cm2 using Shewanella oneidensis as a model biocatalyst without any optimization of bacterial culture. The medium consumption and the response time of the flow-through device are reduced by 16.4 times and 4.2 times, respectively, compared to the non-flow-through counterpart with its freeway space volume six times the volume of graphene foam anode. The graphene foam enabled microfluidic flow-through approach will allow efficient microbial conversion of carbon-containing bioconvertible substrates to electricity with smaller space, less medium consumption, and shorter start-up time. PMID:28120875

  11. Integrated Microfluidic Flow-Through Microbial Fuel Cells

    NASA Astrophysics Data System (ADS)

    Jiang, Huawei; Ali, Md. Azahar; Xu, Zhen; Halverson, Larry J.; Dong, Liang

    2017-01-01

    This paper reports on a miniaturized microbial fuel cell with a microfluidic flow-through configuration: a porous anolyte chamber is formed by filling a microfluidic chamber with three-dimensional graphene foam as anode, allowing nutritional medium to flow through the chamber to intimately interact with the colonized microbes on the scaffolds of the anode. No nutritional media flow over the anode. This allows sustaining high levels of nutrient utilization, minimizing consumption of nutritional substrates, and reducing response time of electricity generation owing to fast mass transport through pressure-driven flow and rapid diffusion of nutrients within the anode. The device provides a volume power density of 745 μW/cm3 and a surface power density of 89.4 μW/cm2 using Shewanella oneidensis as a model biocatalyst without any optimization of bacterial culture. The medium consumption and the response time of the flow-through device are reduced by 16.4 times and 4.2 times, respectively, compared to the non-flow-through counterpart with its freeway space volume six times the volume of graphene foam anode. The graphene foam enabled microfluidic flow-through approach will allow efficient microbial conversion of carbon-containing bioconvertible substrates to electricity with smaller space, less medium consumption, and shorter start-up time.

  12. Automated contrast medium monitoring system for computed tomography--Intra-institutional audit.

    PubMed

    Lauretti, Dario Luca; Neri, Emanuele; Faggioni, Lorenzo; Paolicchi, Fabio; Caramella, Davide; Bartolozzi, Carlo

    2015-12-01

    The aim of this study was to analyze the usage and the data recorded by a RIS-PACS-connected contrast medium (CM) monitoring system (Certegra(®), Bayer Healthcare, Leverkusen, Germany) over 19 months of CT activity. The system used was connected to two dual syringe power injectors (each associated with a 16-row and a high definition 64-row multidetector CT scanner, respectively), allowing to manage contrast medium injection parameters and to send and retrieve CT study-related information via RIS/PACS for any scheduled contrast-enhanced CT examination. The system can handle up to 64 variables and can be accessed via touchscreen by CT operators as well as via a web interface by registered users with three different hierarchy levels. Data related to CM injection parameters (i.e. iodine concentration, volume and flow rate of CM, iodine delivery rate and iodine dose, CM injection pressure, and volume and flow rate of saline), patient weight and height, and type of CT study over a testing period spanning from 1 June 2013 to 10 January 2015 were retrieved from the system. Technical alerts occurred for each injection event (such as system disarm due to technical failure, disarm due to operator's stop, incomplete filling of patient data fields, or excessively high injection pressure), as well as interoperability issues related to data sending and receiving to/from the RIS/PACS were also recorded. During the testing period, the CM monitoring system generated a total of 8609 reports, of which 7629 relative to successful injection events (88.6%). 331 alerts were generated, of which 40 resulted in injection interruption and 291 in CM flow rate limitation due to excessively high injection pressure (>325 psi). Average CM volume and flow rate were 93.73 ± 17.58 mL and 3.53 ± 0.89 mL/s, and contrast injection pressure ranged between 5 and 167 psi. A statistically significant correlation was found between iodine concentration and peak IDR (rs=0.2744, p<0.0001), as well as between iodine concentration and iodine dose (rs=0.3862, p<0.0001) for all CT studies. Automated contrast management systems can provide a full report of contrast use with the possibility to systematically compare different contrast injection protocols, minimize errors, and optimize organ-specific contrast enhancement for any given patient and clinical application. This can be useful to improve and harmonize the quality and consistency of contrast CT procedures within the same radiological department and across the hospital, as well as to monitor potential adverse events and overall costs. Copyright © 2015 Elsevier Ltd. All rights reserved.

  13. Boson mode, Medium Range Structure and Intermediate Phase (IP) in (Na2O)x(B2O3)1-x glasses

    NASA Astrophysics Data System (ADS)

    Vignarooban, K.; Boolchand, P.; Micoulaut, M.; Malki, M.

    2012-02-01

    Raman scattering of titled glasses are examined using a T64000 Dispersive system. Scattering strengths of the Boson mode (40 cm-1, 70 cm-1) and the Boroxyl ring (BR) mode (808 cm-1) are found to decrease with increasing x at the same rate in the 0 < x < 20% soda range. Apparently, the 2D character of BRs embedded in a 3D network gives rise to the Boson mode.ootnotetextM. Flores-Ruiz and G. Naumis, PRB, 2011. 83: p. 184204 The triad of modes (705, 740, 770 cm-1) near the 808 cm-1 mode are found to display a maximum in scattering strength near x = 37% (705 cm-1), 33% (740 cm-1) and 25% (770 cm-1), suggesting that these are also ring modes of Na-tripentaborate (STPB), Na-diborate (SDB) and Na-triborate (STB) super-structures. Variations in Raman scattering strengths also suggest that STB percolate near x = 20%, the stress transition, while the STPB and SDTB percolate near x = 40%, the rigidity transition. These transitions were inferred from m-DSC experiments that show an intermediate phase in the 20% < x < 40% range in dry and homogeneous glasses.

  14. Paracrine Activation of the Wnt/β-Catenin Pathway by Bone Marrow Stem Cell Attenuates Cisplatin-Induced Kidney Injury.

    PubMed

    Jiao, Xiaoyan; Cai, Jieru; Yu, Xiaofang; Ding, Xiaoqiang

    2017-01-01

    Cisplatin-induced acute kidney injury (AKI) involves damage to tubular cells via excess reactive oxygen species (ROS) generation. Stem cell-based therapies have shown great promise in AKI treatment. In this study, we aimed to assess the protective effect and mechanism of bone marrow mesenchymal stem cell (BMSC)-derived conditioned medium (CM) against cisplatin-induced AKI. In vitro, NRK-52E cells were incubated with cisplatin in the presence or absence of CM, followed by the assessment of cell viability, apoptosis and cell cycle distribution. Then, ICG-001 and IWR-1 were used to inhibit the wnt/β-catenin pathway. Furthermore, intracellular and mitochondrial ROS levels were evaluated using DCFH-DA and MitoSOX, respectively. In vivo, after cisplatin injection, rats were intravenously injected with CM or BMSCs. Sera and kidney tissues were collected on day 3 after cisplatin injection to evaluate changes in renal function and histology. Western blotting and qRT-PCR were employed to determine the expression of wnt/β-catenin pathway-related genes and proteins. Immunohistochemical staining was used to evaluate tubular β-catenin expression in kidney biopsy from AKI patients. CM protected NRK-52E cells from cisplatin-induced injury by restoring the wnt4/β-catenin pathway. In response to ICG-001 and IWR-1, the protective effect of CM was attenuated, characterized by a decrease in cell proliferation and an increase in cell apoptosis and intracellular and mitochondrial ROS levels. Knockdown of β-catenin using siRNAs also suppressed the mitochondrial biogenesis regulators PGC-1α, TFAM and NRF-1. In the rat model, CM significantly alleviated renal function and histology associated with tubular injury and upregulated wnt4 and β-catenin. However, the renoprotective effect of CM was blocked by ICG-001, characterized by exacerbated renal function, suppressed PGC-1α expression and increased mitochondrial ROS. Clinical data showed that the tubular β-catenin level was lower in AKI patients experiencing partial recovery than in patients experiencing complete recovery. The activation of the wnt/β-catenin pathway by CM protects against cisplatin-induced kidney injury, resulting in reduced apoptosis and intracellular ROS levels. © 2017 The Author(s). Published by S. Karger AG, Basel.

  15. Regulation of lipoprotein lipase in primary cultures of isolated human adipocytes

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

    Kern, P.A.; Marshall, S.; Eckel, R.H.

    1985-01-01

    To study the regulation of adipose tissue lipoprotein lipase (LPL) in human adipocytes, omental adipose tissue was obtained from healthy subjects and digested in collagenase. The isolated adipocytes thus obtained were suspended in Medium 199 and cultured at 37 degrees C. Cell viability was demonstrated in adipocytes cultured for up to 72 h by constancy of cell number, cell size, trypan-blue exclusion, and specific /sup 125/I-insulin binding. In addition, chloroquine induced an increase in cell-associated /sup 125/I-insulin at 24, 48, and 72 h after preparation. Thus, isolated adipocytes retained their ability to bind, internalize, and degrade insulin. LPL was measuredmore » as activity secreted into the culture medium (CM), released from cells by heparin (HR), and extracted from cell digests. A broad range of heparin concentrations produced a prompt release of LPL from a rapidly replenishable pool of cellular activity. When cells were cultured in medium containing 10% fetal bovine serum, there was a marked stimulation of CM and HR. The secretory response to serum (CM) correlated strongly with HR 24 h after preparation. In addition, HR was found to correlate logarithmically and inversely with body mass index. Insulin, at 400 ng/ml only, increased HR by 36 +/- 10%, an effect simulated by lower concentrations of insulin-like growth factor-1 (IGF1). Thus, LPL is produced and regulated in isolated human adipocytes. The degree of adiposity and serum are important regulators of HR activity, whereas insulin is stimulatory only at a pharmacologic concentration. This effect of insulin may be mediated through the IGF1 receptor. Isolated human adipocytes represent a novel and useful system for the study of LPL and lipid metabolism as well as for other aspects of adipocyte biology.« less

  16. Poly(γ-Glutamic Acid) as an Exogenous Promoter of Chondrogenic Differentiation of Human Mesenchymal Stem/Stromal Cells.

    PubMed

    Antunes, Joana C; Tsaryk, Roman; Gonçalves, Raquel M; Pereira, Catarina Leite; Landes, Constantin; Brochhausen, Christoph; Ghanaati, Shahram; Barbosa, Mário A; Kirkpatrick, C James

    2015-06-01

    Cartilage damage and/or aging effects can cause constant pain, which limits the patient's quality of life. Although different strategies have been proposed to enhance the limited regenerative capacity of cartilage tissue, the full production of native and functional cartilaginous extracellular matrix (ECM) has not yet been achieved. Poly(γ-glutamic acid) (γ-PGA), a naturally occurring polyamino acid, biodegradable into glutamate residues, has been explored for tissue regeneration. In this work, γ-PGA's ability to support the production of cartilaginous ECM by human bone marrow mesenchymal stem/stromal cells (MSCs) and nasal chondrocytes (NCs) was investigated. MSC and NC pellets were cultured in basal medium (BM), chondrogenic medium (CM), and CM-γ-PGA-supplemented medium (CM+γ-PGA) over a period of 21 days. Pellet size/shape was monitored with time. At 14 and 21 days of culture, the presence of sulfated glycosaminoglycans (sGAGs), type II collagen (Col II), Sox-9, aggrecan, type XI collagen (Col XI), type X collagen (Col X), calcium deposits, and type I collagen (Col I) was analyzed. After excluding γ-PGA's cytotoxicity, earlier cell condensation, higher sGAG content, Col II, Sox-9 (day 14), aggrecan, and Col X (day 14) production was observed in γ-PGA-supplemented MSC cultures, with no signs of mineralization or Col I. These effects were not evident with NCs. However, Sox-9 (at day 14) and Col X (at days 14 and 21) were increased, decreased, or absent, respectively. Overall, γ-PGA improved chondrogenic differentiation of MSCs, increasing ECM production earlier in culture. It is proposed that γ-PGA incorporation in novel biomaterials has a beneficial impact on future approaches for cartilage regeneration.

  17. A novel superporous agarose medium for high-speed protein chromatography.

    PubMed

    Shi, Qing-Hong; Zhou, Xin; Sun, Yan

    2005-12-05

    A novel superporous agarose (SA) bead characterized by the presence of wide pores has been fabricated by water-in-oil emulsification using solid granules of calcium carbonate as porogenic agent. After cross-linking, the solid granules were removed by dissolving them in hydrochloric acid. Then, the gel was modified with diethylaminoethyl groups to create an anion exchanger, SA-DEAE, for protein adsorption. A homogeneous agarose (HA) bead was also produced and modified with DEAE for comparison. It was found that the porosity of SA-DEAE was about 6% larger than that of HA-DEAE. Moreover, both optical micrographs and confocal laser scanning microscopy (CLSM) of the ion exchangers with adsorbed fluorescein isothiocyanate (FITC) labeled IgG revealed the superporous structure of the SA medium. In addition, the SA-DEAE column had lower backpressure than the HA-DEAE column, confirming the convective flow of mobile phase through the wide pores. Due to the presence of the wide pores, more channels were available for protein transport and, furthermore, more diffusive pores in the agarose network were accessible for the protein approach from different directions. This led to 40% higher protein capacity and two times higher effective pore diffusivity in the SA-DEAE than in HA-DEAE. Moreover, an increase of the efficiency of the SA-DEAE column until a flow rate of 5 cm/min and the independency of the column efficiency at flow rates from 5 to 17.8 cm/min was found, indicating that intraparticle mass transfer was intensified by convective flow at elevated flow rates. Therefore, the chromatographic resolution of IgG and BSA was little affected up to a flow rate of 17.8 cm/min. The results indicate that the SA medium is favorable for high-speed protein chromatography. (c) 2005 Wiley Periodicals, Inc.

  18. Poly(γ-Glutamic Acid) as an Exogenous Promoter of Chondrogenic Differentiation of Human Mesenchymal Stem/Stromal Cells

    PubMed Central

    Antunes, Joana C.; Tsaryk, Roman; Gonçalves, Raquel M.; Pereira, Catarina Leite; Landes, Constantin; Brochhausen, Christoph; Ghanaati, Shahram

    2015-01-01

    Cartilage damage and/or aging effects can cause constant pain, which limits the patient's quality of life. Although different strategies have been proposed to enhance the limited regenerative capacity of cartilage tissue, the full production of native and functional cartilaginous extracellular matrix (ECM) has not yet been achieved. Poly(γ-glutamic acid) (γ-PGA), a naturally occurring polyamino acid, biodegradable into glutamate residues, has been explored for tissue regeneration. In this work, γ-PGA's ability to support the production of cartilaginous ECM by human bone marrow mesenchymal stem/stromal cells (MSCs) and nasal chondrocytes (NCs) was investigated. MSC and NC pellets were cultured in basal medium (BM), chondrogenic medium (CM), and CM-γ-PGA-supplemented medium (CM+γ-PGA) over a period of 21 days. Pellet size/shape was monitored with time. At 14 and 21 days of culture, the presence of sulfated glycosaminoglycans (sGAGs), type II collagen (Col II), Sox-9, aggrecan, type XI collagen (Col XI), type X collagen (Col X), calcium deposits, and type I collagen (Col I) was analyzed. After excluding γ-PGA's cytotoxicity, earlier cell condensation, higher sGAG content, Col II, Sox-9 (day 14), aggrecan, and Col X (day 14) production was observed in γ-PGA-supplemented MSC cultures, with no signs of mineralization or Col I. These effects were not evident with NCs. However, Sox-9 (at day 14) and Col X (at days 14 and 21) were increased, decreased, or absent, respectively. Overall, γ-PGA improved chondrogenic differentiation of MSCs, increasing ECM production earlier in culture. It is proposed that γ-PGA incorporation in novel biomaterials has a beneficial impact on future approaches for cartilage regeneration. PMID:25760236

  19. Synergistic bactericidal action of phytic acid and sodium chloride against Escherichia coli O157:H7 cells protected by a biofilm.

    PubMed

    Kim, Nam Hee; Rhee, Min Suk

    2016-06-16

    The food industry must prevent the build-up of strong Escherichia coli O157:H7 biofilms in food processing environments. The present study examined the bactericidal action of phytic acid (PA), a natural extract from rice bran and the hulls/peels of legumes, against E. coli O157:H7 biofilms. The synergistic bactericidal effects of PA plus sodium chloride (NaCl) were also examined. E. coli O157:H7 biofilms were allowed for form on stainless steel coupons by culture in both rich (tryptic soy broth, TSB) and minimal (M9) medium at 22°C for 6days. Bacterial cells within biofilms grown in M9 medium were significantly more resistant to PA than those grown in TSB (p<0.05); thus M9 medium was selected for further experiments. The anti-biofilm effect of PA was significantly increased by addition of NaCl (2-4%) (p<0.05); indeed, the combination of 0.4% PA plus 3-4% NaCl completely inactivated E. coli O157:H7 biofilms without recovery (a>6.5logCFU/cm(2) reduction). Neither PA nor NaCl alone were this effective (PA, 1.6-2.7logCFU/cm(2) reduction; NaCl, <0.5logCFU/cm(2) reduction). Confocal laser scanning microscopy images of propidium iodide-treated cells showed that PA (0.4%) plus NaCl (2-4%) had marked membrane permeabilizing effects. These results suggest that a sanitizer that combines these two naturally occurring antimicrobial agents may be useful to food safety managers who encounter thick biofilm formation in food processing environments. Copyright © 2016 Elsevier B.V. All rights reserved.

  20. Effect of environmental and cultural conditions on medium pH and explant growth performance of Douglas-fir ( Pseudotsuga menziesii) shoot cultures

    PubMed Central

    Chen, Chien-Chih; Bates, Rick; Carlson, John

    2015-01-01

    The medium pH level of plant tissue cultures has been shown to be essential to many aspects of explant development and growth. Sensitivity or tolerance of medium pH change in vitro varies according to specific requirements of individual species. The objectives of this study are to 1) determine medium pH change over time in storage conditions and with presence of explants, 2) evaluate the effects of medium pH change on explant growth performance and 3) assess the effects of adding a pH stabilizer, 2-(N-morpholino)ethanesulfonic acid (MES) that is commonly used in Douglas-fir micropropagation medium. Vegetative buds were collected in the spring before breaking dormancy from juvenile and mature donor trees for conducting these evaluations. Medium, with or without MES, was pre-adjusted to five pH levels before adding MES, agar and autoclaving. Medium pH changes and explant growth parameters were measured at eight different incubation times. Overall, MES provided a more stable medium pH, relative to starting pH values, under both light and dark storage conditions as well as with presence of explants. A general trend of decreasing medium pH over time was found comparing explants from juvenile and mature donor genotypes. Explant height and weight growth increased over time, but differ among explants from juvenile and mature donor genotypes. Our findings suggest that a 21-day subculture practice may best sustain medium freshness, medium pH level and desirable explant growth. PMID:26535110

  1. Antitumor Activity of Rat Mesenchymal Stem Cells during Direct or Indirect Co-Culturing with C6 Glioma Cells.

    PubMed

    Gabashvili, A N; Baklaushev, V P; Grinenko, N F; Mel'nikov, P A; Cherepanov, S A; Levinsky, A B; Chehonin, V P

    2016-02-01

    The tumor-suppressive effect of rat mesenchymal stem cells against low-differentiated rat C6 glioma cells during their direct and indirect co-culturing and during culturing of C6 glioma cells in the medium conditioned by mesenchymal stem cells was studied in an in vitro experiment. The most pronounced antitumor activity of mesenchymal stem cells was observed during direct co-culturing with C6 glioma cells. The number of live C6 glioma cells during indirect co-culturing and during culturing in conditioned medium was slightly higher than during direct co-culturing, but significantly differed from the control (C6 glioma cells cultured in medium conditioned by C6 glioma cells). The cytotoxic effect of medium conditioned by mesenchymal stem cells was not related to medium depletion by glioma cells during their growth. The medium conditioned by other "non-stem" cells (rat astrocytes and fibroblasts) produced no tumor-suppressive effect. Rat mesenchymal stem cells, similar to rat C6 glioma cells express connexin 43, the main astroglial gap junction protein. During co-culturing, mesenchymal stem cells and glioma C6 cells formed functionally active gap junctions. Gap junction blockade with connexon inhibitor carbenoxolone attenuated the antitumor effect observed during direct co-culturing of C6 glioma cells and mesenchymal stem cells to the level produced by conditioned medium. Cell-cell signaling mediated by gap junctions can be a mechanism of the tumor-suppressive effect of mesenchymal stem cells against C6 glioma cells. This phenomenon can be used for the development of new methods of cell therapy for high-grade malignant gliomas.

  2. A RELATION BETWEEN THE WARM NEUTRAL AND IONIZED MEDIA OBSERVED IN THE CANADIAN GALACTIC PLANE SURVEY

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

    Foster, T.; Kothes, R.; Brown, J. C., E-mail: Tyler.Foster@nrc-cnrc.gc.ca

    2013-08-10

    We report on a comparison between 21 cm rotation measure (RM) and the optically thin atomic hydrogen column density (N{sub H{sub I}}({tau} {yields} 0)) measured toward unresolved extragalactic sources in the Galactic plane of the northern sky. H I column densities integrated to the Galactic edge are measured immediately surrounding each of nearly 2000 sources in 1 arcmin 21 cm line data, and are compared to RMs observed from polarized emission of each source. RM data are binned in column density bins 4 Multiplication-Sign 10{sup 20} cm{sup -2} wide, and one observes a strong relationship between the number of hydrogenmore » atoms in a 1 cm{sup 2} column through the plane and the mean RM along the same line of sight and path length. The relationship is linear over one order of magnitude (from 0.8 to 14 Multiplication-Sign 10{sup 21} atoms cm{sup -2}) of column densities, with a constant RM/N{sub H{sub I}}{approx} -23.2 {+-} 2.3 rad m{sup -2}/10{sup 21} atoms cm{sup -2}, and a positive RM of 45.0 {+-} 13.8 rad m{sup -2} in the presence of no atomic hydrogen. This slope is used to calculate a mean volume-averaged magnetic field in the second quadrant of (B{sub Parallel-To }) {approx}1.0 {+-} 0.1 {mu}G directed away from the Sun, assuming an ionization fraction of 8% (consistent with the warm-neutral medium; WNM). The remarkable consistency between this field and (B) = 1.2 {mu}G found with the same RM sources and a Galactic model of dispersion measures (DMs) suggests that electrons in the partially ionized WNM are mainly responsible for pulsar DMs, and thus the partially ionized WNM is the dominant form of the magneto-ionic interstellar medium.« less

  3. Metabolic activity of odontoblast-like cells irradiated with blue LED (455 nm).

    PubMed

    de Almeida, Leopoldina Fátima Dantas; Basso, Fernanda Gonçalves; Turrioni, Ana Paula Silveira; de-Souza-Costa, Carlos Alberto; Hebling, Josimeri

    2016-01-01

    Blue light emitting diodes (LEDs) are frequently used in dentistry for light activation of resin-based materials; however, their photobiostimulatory effects have not yet been fully investigated. This study aimed to investigate the effect of blue LED (455 nm) on the metabolism of odontoblast-like cells MDPC-23. Energy doses of 2 and 4 J/cm(2) were used at 20 mW/cm(2) fixed power density. MDPC-23 cells were seeded at 10,000 cells/cm(2) density in Dulbecco's modified Eagle's medium (DMEM) containing 10 % fetal bovine serum (FBS). After 12 h, the culture medium was replaced with new DMEM supplemented with 0.5 % of FBS, and the cells were incubated for further 12 h. After that, single irradiation was performed to the culture, under selected parameters. Cell viability evaluations (Alamar Blue Assay, n = 12), number of viable cells (Trypan Blue Assay, n = 12), morphological analysis by scanning electron microscopy (SEM, n = 2), gene expression (n = 6) of alkaline phosphatase (Alp), collagen (Col-1a1), and dental matrix protein (Dmp-1) (quantitative polymerase chain reaction (qPCR)) were performed 72 h after irradiation. Data were analyzed by Kruskal-Wallis, ANOVA, and Tukey tests (p < 0.05). Direct light application at 4 J/cm(2) energy dose had no negative effects on cell viability, while irradiation with 2 J/cm(2) reduced cell metabolism. None of doses affected the number of viable cells compared with the control group. The two energy doses downregulated the expression of Alp; however, expression of Col-1a1 and Dmp-1 had no alteration. Cells presented change in the cytoskeleton only when irradiated with 2 J/cm(2). In conclusion, the blue LED (455 nm) irradiation, under the evaluated parameters, had no biostimulatory effects on MDPC-23 cells.

  4. BENZENE FORMATION ON INTERSTELLAR ICY MANTLES CONTAINING PROPARGYL ALCOHOL

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

    Sivaraman, B.; Mukherjee, R.; Subramanian, K. P.

    Propargyl alcohol (CHCCH{sub 2}OH) is a known stable isomer of the propenal (CH{sub 2}CHCHO) molecule that was reported to be present in the interstellar medium (ISM). At astrochemical conditions in the laboratory, icy layers of propargyl alcohol grown at 85 K were irradiated by 2 keV electrons and probed by a Fourier Transform InfraRed spectrometer in the mid-infrared (IR) region, 4000-500 cm{sup –1}. Propargyl alcohol ice under astrochemical conditions was studied for the first time; therefore, IR spectra of reported amorphous (85 K) and crystalline (180 K) propargyl alcohol ices can be used to detect its presence in the ISM.more » Moreover, our experiments clearly show benzene (C{sub 6}H{sub 6}) formation to be the major product from propargyl alcohol irradiation, confirming the role of propargyl radicals (C{sub 3}H{sub 3}) formed from propargyl alcohol dissociation that was long expected based on theoretical modeling to effectively synthesize C{sub 6}H{sub 6} in the interstellar icy mantles.« less

  5. Copper oxide assisted cysteine hierarchical structures for immunosensor application

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

    Pandey, Chandra Mouli; Department of Chemistry, Faculty of Science, Banaras Hindu University, Varanasi 221005; Sumana, Gajjala, E-mail: sumanagajjala@gmail.com

    2014-09-08

    The present work describes the promising electrochemical immunosensing strategy based on copper (II) assisted hierarchical cysteine structures (CuCys) varying from star to flower like morphology. The CuCys having average size of 10 μm have been synthesised using L-Cysteine as initial precursor in presence of copper oxide under environmentally friendly conditions in aqueous medium. To delineate the synthesis mechanism, detailed structural investigations have been carried out using characterization techniques such as X-ray diffraction, transmission electron microscopy, and Fourier transform infrared spectroscopy. The electrochemical behaviour of self-assembled CuCys on gold electrode shows surface controlled electrode reaction with an apparent electron transfer rate constantmore » of 3.38 × 10{sup −4 }cm s{sup −1}. This innovative platform has been utilized to fabricate an immunosensor by covalently immobilizing monoclonal antibodies specific for Escherichia coli O157:H7 (E. coli). Under the optimal conditions, the fabricated immunosensor is found to be sensitive and specific for the detection of E. coli with a detection limit of 10 cfu/ml.« less

  6. [In vitro flowering of cultures from a hybrid of Cymbidium goeringii and C. hybridium].

    PubMed

    Zheng, Li-Ming; Pang, Ji-Liang

    2006-06-01

    Wild-type female spring orchid (Cymbidium goeringii) was crossed with male Cymbidium hybridium. Over eight hundred protocorm clones were obtained from hybrid offsprings. Among them, one protocorm clone was identified to differentiate visible floral buds two months after subculture in vitro (Plate I). The protocorms and shoots derived from this clone were further used in studying the effects of abscisic acid (ABA) and paclobutrazol (PP333) pretreatment as well as different concentrations of 6-benzyladenine (6-BA) on floral bud differentiation. The optimum combination of hormones in floral bud induction was 6-BA 1.0 mg/L and NAA 0.1 mg/L, and total frequency of floral bud formation was up to 31% (Table 1). The optimum length of shoots used in floral bud induction was 1-2 cm, and the frequency of floral bud formation was 19% (Table 1). The increase in total frequency was not significant in floral bud induction from protocorms and shoots with length of 1-2 cm or 2-4 cm cultured on MS medium containing 6-BA 1.0 mg/L and NAA 0.1 mg/L after pretreatment on MS medium supplemented with ABA 0.5 mg/L and PP333 0.5 mg/L for 35 d (Table 2).

  7. THE INTERSTELLAR MEDIUM IN THE KEPLER SEARCH VOLUME

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

    Johnson, Marshall C.; Redfield, Seth; Jensen, Adam G., E-mail: mjohnson@astro.as.utexas.edu

    2015-07-10

    The properties of the interstellar medium (ISM) surrounding a planetary system can impact planetary climate through a number of mechanisms, including changing the size of the astrosphere (one of the major shields for cosmic rays) as well as direct deposition of material into planetary atmospheres. In order to constrain the ambient ISM conditions for exoplanetary systems, we present observations of interstellar Na i and K i absorption toward seventeen early type stars in the Kepler prime mission field of view (FOV). We identify 39 Na i and 8 K i velocity components, and attribute these to 11 ISM clouds. Sixmore » of these are detected toward more than one star, and for these clouds we put limits on the cloud properties, including distance and hydrogen number density. We identify one cloud with significant (≳1.5 cm{sup −3}) hydrogen number density located within the nominal ∼100 pc boundary of the Local Bubble. We identify systems with confirmed planets within the Kepler FOV that could lie within these ISM clouds, and estimate upper limits on the astrosphere sizes of these systems under the assumption that they do lie within these clouds. Under this condition, the Kepler-20, 42, and 445 multiplanet systems could have compressed astrospheres much smaller than the present-day heliosphere. Among the known habitable zone planet hosts, Kepler-186 could have an astrosphere somewhat smaller than the heliosphere, while Kepler-437 and KOI-4427 could have astrospheres much larger than the heliosphere. The thick disk star Kepler-444 may have an astrosphere just a few AU in radius.« less

  8. Flow through internal elastic lamina affects shear stress on smooth muscle cells (3D simulations).

    PubMed

    Tada, Shigeru; Tarbell, John M

    2002-02-01

    We describe a three-dimensional numerical simulation of interstitial flow through the medial layer of an artery accounting for the complex entrance condition associated with fenestral pores in the internal elastic lamina (IEL) to investigate the fluid mechanical environment around the smooth muscle cells (SMCs) right beneath the IEL. The IEL was modeled as an impermeable barrier to water flow except for the fenestral pores, which were assumed to be uniformly distributed over the IEL. The medial layer was modeled as a heterogeneous medium composed of a periodic array of cylindrical SMCs embedded in a continuous porous medium representing the interstitial proteoglycan and collagen matrix. Depending on the distance between the IEL bottom surface and the upstream end of the proximal layer of SMCs, the local shear stress on SMCs right beneath the fenestral pore could be more than 10 times higher than that on the cells far removed from the IEL under the conditions that the fenestral pore diameter and area fraction of pores were kept constant at 1.4 microm and 0.05, respectively. Thus these proximal SMCs may experience shear stress levels that are even higher than endothelial cells exposed to normal blood flow (order of 10 dyn/cm(2)). Furthermore, entrance flow through fenestral pores alters considerably the interstitial flow field in the medial layer over a spatial length scale of the order of the fenestral pore diameter. Thus the spatial gradient of shear stress on the most superficial SMC is noticeably higher than computed for endothelial cell surfaces.

  9. Humoral Activity of Cord Blood-Derived Stem/Progenitor Cells: Implications for Stem Cell-Based Adjuvant Therapy of Neurodegenerative Disorders

    PubMed Central

    Paczkowska, Edyta; Kaczyńska, Katarzyna; Pius-Sadowska, Ewa; Rogińska, Dorota; Kawa, Miłosz; Ustianowski, Przemysław; Safranow, Krzysztof; Celewicz, Zbigniew; Machaliński, Bogusław

    2013-01-01

    Background Stem/progenitor cells (SPCs) demonstrate neuro-regenerative potential that is dependent upon their humoral activity by producing various trophic factors regulating cell migration, growth, and differentiation. Herein, we compared the expression of neurotrophins (NTs) and their receptors in specific umbilical cord blood (UCB) SPC populations, including lineage-negative, CD34+, and CD133+ cells, with that in unsorted, nucleated cells (NCs). Methods and Results The expression of NTs and their receptors was detected by QRT-PCR, western blotting, and immunofluorescent staining in UCB-derived SPC populations (i.e., NCs vs. lineage-negative, CD34+, and CD133+ cells). To better characterize, global gene expression profiles of SPCs were determined using genome-wide RNA microarray technology. Furthermore, the intracellular production of crucial neuro-regenerative NTs (i.e., BDNF and NT-3) was assessed in NCs and lineage-negative cells after incubation for 24, 48, and 72 h in both serum and serum-free conditions. We discovered significantly higher expression of NTs and NT receptors at both the mRNA and protein level in lineage-negative, CD34+, and CD133+ cells than in NCs. Global gene expression analysis revealed considerably higher expression of genes associated with the production and secretion of proteins, migration, proliferation, and differentiation in lineage-negative cells than in CD34+ or CD133+ cell populations. Notably, after short-term incubation under serum-free conditions, lineage-negative cells and NCs produced significantly higher amounts of BDNF and NT-3 than under steady-state conditions. Finally, conditioned medium (CM) from lineage-negative SPCs exerted a beneficial impact on neural cell survival and proliferation. Conclusions Collectively, our findings demonstrate that UCB-derived SPCs highly express NTs and their relevant receptors under steady-state conditions, NT expression is greater under stress-related conditions and that CM from SPCs favorable influence neural cell proliferation and survival. Understanding the mechanisms governing the characterization and humoral activity of subsets of SPCs may yield new therapeutic strategies that might be more effective in treating neurodegenerative disorders. PMID:24391835

  10. Humoral activity of cord blood-derived stem/progenitor cells: implications for stem cell-based adjuvant therapy of neurodegenerative disorders.

    PubMed

    Paczkowska, Edyta; Kaczyńska, Katarzyna; Pius-Sadowska, Ewa; Rogińska, Dorota; Kawa, Miłosz; Ustianowski, Przemysław; Safranow, Krzysztof; Celewicz, Zbigniew; Machaliński, Bogusław

    2013-01-01

    Stem/progenitor cells (SPCs) demonstrate neuro-regenerative potential that is dependent upon their humoral activity by producing various trophic factors regulating cell migration, growth, and differentiation. Herein, we compared the expression of neurotrophins (NTs) and their receptors in specific umbilical cord blood (UCB) SPC populations, including lineage-negative, CD34(+), and CD133(+) cells, with that in unsorted, nucleated cells (NCs). The expression of NTs and their receptors was detected by QRT-PCR, western blotting, and immunofluorescent staining in UCB-derived SPC populations (i.e., NCs vs. lineage-negative, CD34(+), and CD133(+) cells). To better characterize, global gene expression profiles of SPCs were determined using genome-wide RNA microarray technology. Furthermore, the intracellular production of crucial neuro-regenerative NTs (i.e., BDNF and NT-3) was assessed in NCs and lineage-negative cells after incubation for 24, 48, and 72 h in both serum and serum-free conditions. We discovered significantly higher expression of NTs and NT receptors at both the mRNA and protein level in lineage-negative, CD34(+), and CD133(+) cells than in NCs. Global gene expression analysis revealed considerably higher expression of genes associated with the production and secretion of proteins, migration, proliferation, and differentiation in lineage-negative cells than in CD34(+) or CD133(+) cell populations. Notably, after short-term incubation under serum-free conditions, lineage-negative cells and NCs produced significantly higher amounts of BDNF and NT-3 than under steady-state conditions. Finally, conditioned medium (CM) from lineage-negative SPCs exerted a beneficial impact on neural cell survival and proliferation. Collectively, our findings demonstrate that UCB-derived SPCs highly express NTs and their relevant receptors under steady-state conditions, NT expression is greater under stress-related conditions and that CM from SPCs favorable influence neural cell proliferation and survival. Understanding the mechanisms governing the characterization and humoral activity of subsets of SPCs may yield new therapeutic strategies that might be more effective in treating neurodegenerative disorders.

  11. Anaerobic digestion of citrus waste using two-stage membrane bioreactor

    NASA Astrophysics Data System (ADS)

    Millati, Ria; Lukitawesa; Dwi Permanasari, Ervina; Wulan Sari, Kartika; Nur Cahyanto, Muhammad; Niklasson, Claes; Taherzadeh, Mohammad J.

    2018-03-01

    Anaerobic digestion is a promising method to treat citrus waste. However, the presence of limonene in citrus waste inhibits anaerobic digestion process. Limonene is an antimicrobial compound and could inhibit methane forming bacteria that takes a longer time to recover than the injured acid forming bacteria. Hence, volatile fatty acids will be accumulated and methane production will be decreased. One way to solve this problem is by conducting anaerobic digestion process into two stages. The first step is aimed for hydrolysis, acidogenesis, and acetogenesis reactions and the second stage is aimed for methanogenesis reaction. The separation of the system would further allow each stage in their optimum conditions making the process more stable. In this research, anaerobic digestion was carried out in batch operations using 120 ml-glass bottle bioreactors in 2 stages. The first stage was performed in free-cells bioreactor, whereas the second stage was performed in both bioreactor of free cells and membrane bioreactor. In the first stage, the reactor was set into ‘anaerobic’ and ‘semi-aerobic’ conditions to examine the effect of oxygen on facultative anaerobic bacteria in acid production. In the second stage, the protection of membrane towards the cells against limonene was tested. For the first stage, the basal medium was prepared with 1.5 g VS of inoculum and 4.5 g VS of citrus waste. The digestion process was carried out at 55°C for four days. For the second stage, the membrane bioreactor was prepared with 3 g of cells that were encased and sealed in a 3×6 cm2 polyvinylidene fluoride membrane. The medium contained 40 ml basal medium and 10 ml liquid from the first stage. The bioreactors were incubated at 55°C for 2 days under anaerobic condition. The results from the first stage showed that the maximum total sugar under ‘anaerobic’ and ‘semi-aerobic’ conditions was 294.3 g/l and 244.7 g/l, respectively. The corresponding values for total volatile fatty acids were 3.8 g/l and 2.9 g/l, respectively. Methane production of citrus waste taken from the first stage under ‘anaerobic’ condition in membrane and free-cells bioreactors was 11.2 Nml and 7.2 Nml, respectively. Whereas, methane production of citrus waste taken from the first stage under ‘semi-aerobic’ condition in membrane and free-cells bioreactors was 8.8 Nml and 5.7 Nml, respectively. It can be seen from the results of the first stage that volatile fatty acids from ‘anaerobic’ condition was higher than that of ‘semi-aerobic’ condition. The absence of oxygen provides the optimal condition for growth and metabolism of facultative and obligatorily anaerobic bacteria in the first stage. Furthermore, polyvinylidene fluoride membrane was able to protect the cells from antimicrobial compounds.

  12. [The study of the role of the water medium in the mechanism of action of peptides in low and ultra low doses].

    PubMed

    Grigor'ev, E I; Khavinson, V Kh; Malinin, V V; Grigor'ev, A E; Kochnev, I N; Kudriavtseva, T A

    2003-01-01

    The correlation between the structures and conformations of short peptides KE, EW, AEDG and other, their influence on the dynamic properties of water and dose/biologic effect dependencies in a wide range of concentrations were regarded. Their effects on the dynamic properties of water were studied by temperature dependencies (5-45 degrees C) of infrared spectra of the solutions in the near (5180 cm-1) and far (200 cm-1). In vitro biotesting included the determination of the proliferative activity of thymocytes, a bimodal curve with the second maximum were detected at super-low doses (10(-17)-10(-15) mol/l). Authors propose a hypothesis that for superlow concentrations the formation and distance transmission of a signal from ligand to a target cell without the formation of any ligand-receptor complex take place. An active role in this model belongs to water medium acting according to the solution mechanism.

  13. Extreme ultraviolet observations of G191-B2B and the local interstellar medium with the Hopkins Ultraviolet Telescope

    NASA Technical Reports Server (NTRS)

    Kimble, Randy A.; Davidsen, Arthur F.; Blair, William P.; Bowers, Charles W.; Van Dyke Dixon, W.; Durrance, Samuel T.; Feldman, Paul D.; Ferguson, Henry C.; Henry, Richard C.; Kriss, Gerard A.

    1993-01-01

    During the Astro-l mission in 1990 December, the Hopkins Ultraviolet Telescope (HUT) was used to observe the extreme ultraviolet spectrum (415-912 A) of the hot DA white dwarf GI91-B2B. Absorption by neutral helium shortward of the 504 A He I absorption edge is clearly detected in the raw spectrum. Model fits to the observed spectrum require interstellar neutral helium and neutral hydrogen column densities of 1.45 +/- 0.065 x 10 exp 17/sq cm and 1.69 +/- 0.12 x 10 exp 18/sq cm, respectively. Comparison of the neutral columns yields a direct assessment of the ionization state of the local interstellar cloud surrounding the Sun. The neutral hydrogen to helium ratio of 11.6 +/- 1.0 observed by HUT strongly contradicts the widespread view that hydrogen is much more ionized than helium in the local interstellar medium, a view which has motivated some exotic theoretical explanations for the supposed high ionization.

  14. The yeast Scheffersomyces amazonensis is an efficient xylitol producer.

    PubMed

    Cadete, Raquel M; Melo-Cheab, Monaliza A; Viana, Adriana L; Oliveira, Evelyn S; Fonseca, César; Rosa, Carlos A

    2016-12-01

    This study assessed the efficiency of Scheffersomyces amazonensis UFMG-CM-Y493 T , cultured in xylose-supplemented medium (YPX) and rice hull hydrolysate (RHH), to convert xylose to xylitol under moderate and severe oxygen limitation. The highest xylitol yields of 0.75 and 1.04 g g -1 in YPX and RHH, respectively, were obtained under severe oxygen limitation. However, volumetric productivity in RHH was ninefold decrease than that in YPX medium. The xylose reductase (XR) and xylitol dehydrogenase (XDH) activities in the YPX cultures were strictly dependent on NADPH and NAD + respectively, and were approximately 10% higher under severe oxygen limitation than under moderate oxygen limitation. This higher xylitol production observed under severe oxygen limitation can be attributed to the higher XR activity and shortage of the NAD + needed by XDH. These results suggest that Sc. amazonensis UFMG-CM-Y493 T is one of the greatest xylitol producers described to date and reveal its potential use in the biotechnological production of xylitol.

  15. Electrochemical degradation of the antihypertensive losartan in aqueous medium by electro-oxidation with boron-doped diamond electrode.

    PubMed

    Salazar, Claudio; Contreras, Nicole; Mansilla, Héctor D; Yáñez, Jorge; Salazar, Ricardo

    2016-12-05

    In this work the electrochemical oxidation of losartan, an emerging pharmaceutical pollutant, was studied. Electrochemical oxidation was carried out in batch mode, in an open and undivided cell of 100cm(3) using a boron-doped diamond (BDD)/stainless steel system. With Cl(-) medium 56% of mineralization was registered, while with the trials containing SO4(2-) as supporting electrolyte a higher mineralization yield of 67% was reached, even obtaining a total removal of losartan potassium at 80mAcm(-2) and 180min of reaction time at pH 7.0. Higher losartan potassium concentrations enhanced the mineralization degree and the efficiency of the electrochemical oxidation process. During the mineralization up to 4 aromatic intermediates were identified by ultra high performance liquid chromatography tandem mass spectrometry (UHPLC-MS/MS). Moreover, short-linear carboxylic acids, like oxalic, succinic and oxamic were detected and quantified by ion-exclusion HPLC. Finally, the ability of the electrochemical oxidation process to mineralize dissolved commercial tablets containing losartan was achieved, obtaining TOC removal up to 71% under optimized conditions (10mAcm(-2), 0.05M Na2SO4, pH 7.0 and 25°C and 360min of electrolysis). Copyright © 2016 Elsevier B.V. All rights reserved.

  16. X-irradiation of human bronchial cancer cells causes the bystander effects in normal bronchial cells in vitro.

    PubMed

    Konopacka, M; Rogoliński, J

    2010-01-01

    Using X radiation commonly used in radiotherapy of cancers we investigated bystander interactions between human cells: irradiated A549 bronchial carcinoma human cells and non irradiated BEAS-2B normal bronchial epithelial cells. Non irradiated cells were incubated in medium transferred from irradiated A549 cells (ICM-irradiation conditioned medium) for 48h and next the chromosomal damage and apoptosis were estimated. Conditioned medium collected from irradiated cancer cells induced in non irradiated cells of the same line as well as in BEAS-2B normal cells genetic changes such as micronuclei, chromatid and chromosomal breaks and condensation of chromatin characteristic for processes of apoptosis. Addition of only 1% of conditioned medium to fresh medium was sufficient to induction of bystander response to normal bronchial cells. The presented results in this study could have implications for human radiation risk and in evaluating the secondary effects of radiotherapy.

  17. Bright X-ray transient in the LMC

    NASA Astrophysics Data System (ADS)

    Saxton, R.; Read, A. M.; Li, D. Y.

    2018-01-01

    We report a bright X-ray transient in the LMC from an XMM-Newton slew made on 5th January 2018. The source, XMMSL2 J053629.4-675940, had a soft X-ray (0.2-2 keV) count rate in the EPIC-pn detector, medium filter of 1.82+/-0.56 c/s, equivalent to a flux Fx=2.3+/-0.7E-12 ergs/s/cm2 for a nominal spectrum of a power-law of slope 2 absorbed by a column NH=3E20 cm^-2.

  18. The local interstellar helium density - Corrected

    NASA Technical Reports Server (NTRS)

    Freeman, J.; Paresce, F.; Bowyer, S.

    1979-01-01

    An upper bound for the number density of neutral helium in the local interstellar medium of 0.004 + or - 0.0022 per cu cm was previously reported, based on extreme-ultraviolet telescope observations at 584 A made during the 1975 Apollo-Soyuz Test Project. A variety of evidence is found which indicates that the 584-A sensitivity of the instrument declined by a factor of 2 between the last laboratory calibration and the time of the measurements. The upper bound on the helium density is therefore revised to 0.0089 + or - 0.005 per cu cm.

  19. An invisible medium for circularly polarized electromagnetic waves.

    PubMed

    Tamayama, Y; Nakanishi, T; Sugiyama, K; Kitano, M

    2008-12-08

    We study the no reflection condition for a planar boundary between vacuum and an isotropic chiral medium. In general chiral media, elliptically polarized waves incident at a particular angle satisfy the no reflection condition. When the wave impedance and wavenumber of the chiral medium are equal to the corresponding parameters of vacuum, one of the circularly polarized waves is transmitted to the medium without reflection or refraction for all angles of incidence. We propose a circular polarizing beam splitter as a simple application of the no reflection effect. (c) 2008 Optical Society of America

  20. Antimicrobial peptides secreted by equine mesenchymal stromal cells inhibit the growth of bacteria commonly found in skin wounds.

    PubMed

    Harman, Rebecca M; Yang, Steven; He, Megan K; Van de Walle, Gerlinde R

    2017-07-04

    The prevalence of chronic skin wounds in humans is high, and treatment is often complicated by the presence of pathogenic bacteria. Therefore, safe and innovative treatments to reduce the bacterial load in cutaneous wounds are needed. Mesenchymal stromal cells (MSC) are known to provide paracrine signals that act on resident skin cells to promote wound healing, but their potential antibacterial activities are not well described. The present study was designed to examine the antibacterial properties of MSC from horses, as this animal model offers a readily translatable model for MSC therapies in humans. Specifically, we aimed to (i) evaluate the in vitro effects of equine MSC on the growth of representative gram-negative and gram-positive bacterial species commonly found in skin wounds and (ii) define the mechanisms by which MSC inhibit bacterial growth. MSC were isolated from the peripheral blood of healthy horses. Gram-negative E. coli and gram-positive S. aureus were cultured in the presence of MSC and MSC conditioned medium (CM), containing all factors secreted by MSC. Bacterial growth was measured by plating bacteria and counting viable colonies or by reading the absorbance of bacterial cultures. Bacterial membrane damage was detected by incorporation of N-phenyl-1-naphthylamine (NPN). Antimicrobial peptide (AMP) gene and protein expression by equine MSC were determined by RT-PCR and Western blot analysis, respectively. Blocking of AMP activity of MSC CM was achieved using AMP-specific antibodies. We found that equine MSC and MSC CM inhibit the growth of E. coli and S. aureus, and that MSC CM depolarizes the cell membranes of these bacteria. In addition, we found that equine MSC CM contains AMPs, and blocking these AMPs with antibodies reduces the effects of MSC CM on bacteria. Our results demonstrate that equine MSC inhibit bacterial growth and secrete factors that compromise the membrane integrity of bacteria commonly found in skin wounds. We also identified four specific AMPs produced by equine MSC. The secretion of AMPs may contribute to the value of MSC as a therapy for cutaneous wounds in both horses and humans.

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