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Sample records for affects cell morphology

  1. Glycopeptidolipid of Mycobacterium smegmatis J15cs Affects Morphology and Survival in Host Cells

    PubMed Central

    Fujiwara, Nagatoshi; Maeda, Shinji; Naka, Takashi; Taniguchi, Hatsumi; Yamamoto, Saburo; Ayata, Minoru

    2015-01-01

    Mycobacterium smegmatis has been widely used as a mycobacterial infection model. Unlike the M. smegmatis mc2155 strain, M. smegmatis J15cs strain has the advantage of surviving for one week in murine macrophages. In our previous report, we clarified that the J15cs strain has deleted apolar glycopeptidolipids (GPLs) in the cell wall, which may affect its morphology and survival in host cells. In this study, the gene causing the GPL deletion in the J15cs strain was identified. The mps1-2 gene (MSMEG_0400-0402) correlated with GPL biosynthesis. The J15cs strain had 18 bps deleted in the mps1 gene compared to that of the mc2155 strain. The mps1-complemented J15cs mutant restored the expression of GPLs. Although the J15cs strain produces a rough and dry colony, the colony morphology of this mps1-complement was smooth like the mc2155 strain. The length in the mps1-complemented J15cs mutant was shortened by the expression of GPLs. In addition, the GPL-restored J15cs mutant did not survive as long as the parent J15cs strain in the murine macrophage cell line J774.1 cells. The results are direct evidence that the deletion of GPLs in the J15cs strain affects bacterial size, morphology, and survival in host cells. PMID:25970481

  2. Aircraft noise exposure affects rat behavior, plasma norepinephrine levels, and cell morphology of the temporal lobe.

    PubMed

    Di, Guo-Qing; Zhou, Bing; Li, Zheng-Guang; Lin, Qi-Li

    2011-12-01

    In order to investigate the physiological effects of airport noise exposure on organisms, in this study, we exposed Sprague-Dawley rats in soundproof chambers to previously recorded aircraft-related noise for 65 d. For comparison, we also used unexposed control rats. Noise was arranged according to aircraft flight schedules and was adjusted to its weighted equivalent continuous perceived noise levels (L(WECPN)) of 75 and 80 dB for the two experimental groups. We examined rat behaviors through an open field test and measured the concentrations of plasma norepinephrine (NE) by high performance liquid chromatography-fluorimetric detection (HPLC-FLD). We also examined the morphologies of neurons and synapses in the temporal lobe by transmission electron microscopy (TEM). Our results showed that rats exposed to airport noise of 80 dB had significantly lower line crossing number (P<0.05) and significantly longer center area duration (P<0.05) than control animals. After 29 d of airport noise exposure, the concentration of plasma NE of exposed rats was significantly higher than that of the control group (P<0.05). We also determined that the neuron and synapsis of the temporal lobe of rats showed signs of damage after aircraft noise of 80 dB exposure for 65 d. In conclusion, exposing rats to long-term aircraft noise affects their behaviors, plasma NE levels, and cell morphology of the temporal lobe.

  3. MeCP2 Affects Skeletal Muscle Growth and Morphology through Non Cell-Autonomous Mechanisms.

    PubMed

    Conti, Valentina; Gandaglia, Anna; Galli, Francesco; Tirone, Mario; Bellini, Elisa; Campana, Lara; Kilstrup-Nielsen, Charlotte; Rovere-Querini, Patrizia; Brunelli, Silvia; Landsberger, Nicoletta

    2015-01-01

    Rett syndrome (RTT) is an autism spectrum disorder mainly caused by mutations in the X-linked MECP2 gene and affecting roughly 1 out of 10.000 born girls. Symptoms range in severity and include stereotypical movement, lack of spoken language, seizures, ataxia and severe intellectual disability. Notably, muscle tone is generally abnormal in RTT girls and women and the Mecp2-null mouse model constitutively reflects this disease feature. We hypothesized that MeCP2 in muscle might physiologically contribute to its development and/or homeostasis, and conversely its defects in RTT might alter the tissue integrity or function. We show here that a disorganized architecture, with hypotrophic fibres and tissue fibrosis, characterizes skeletal muscles retrieved from Mecp2-null mice. Alterations of the IGF-1/Akt/mTOR pathway accompany the muscle phenotype. A conditional mouse model selectively depleted of Mecp2 in skeletal muscles is characterized by healthy muscles that are morphologically and molecularly indistinguishable from those of wild-type mice raising the possibility that hypotonia in RTT is mainly, if not exclusively, mediated by non-cell autonomous effects. Our results suggest that defects in paracrine/endocrine signaling and, in particular, in the GH/IGF axis appear as the major cause of the observed muscular defects. Remarkably, this is the first study describing the selective deletion of Mecp2 outside the brain. Similar future studies will permit to unambiguously define the direct impact of MeCP2 on tissue dysfunctions.

  4. MeCP2 Affects Skeletal Muscle Growth and Morphology through Non Cell-Autonomous Mechanisms

    PubMed Central

    Galli, Francesco; Tirone, Mario; Bellini, Elisa; Campana, Lara; Kilstrup-Nielsen, Charlotte; Rovere-Querini, Patrizia; Brunelli, Silvia; Landsberger, Nicoletta

    2015-01-01

    Rett syndrome (RTT) is an autism spectrum disorder mainly caused by mutations in the X-linked MECP2 gene and affecting roughly 1 out of 10.000 born girls. Symptoms range in severity and include stereotypical movement, lack of spoken language, seizures, ataxia and severe intellectual disability. Notably, muscle tone is generally abnormal in RTT girls and women and the Mecp2-null mouse model constitutively reflects this disease feature. We hypothesized that MeCP2 in muscle might physiologically contribute to its development and/or homeostasis, and conversely its defects in RTT might alter the tissue integrity or function. We show here that a disorganized architecture, with hypotrophic fibres and tissue fibrosis, characterizes skeletal muscles retrieved from Mecp2-null mice. Alterations of the IGF-1/Akt/mTOR pathway accompany the muscle phenotype. A conditional mouse model selectively depleted of Mecp2 in skeletal muscles is characterized by healthy muscles that are morphologically and molecularly indistinguishable from those of wild-type mice raising the possibility that hypotonia in RTT is mainly, if not exclusively, mediated by non-cell autonomous effects. Our results suggest that defects in paracrine/endocrine signaling and, in particular, in the GH/IGF axis appear as the major cause of the observed muscular defects. Remarkably, this is the first study describing the selective deletion of Mecp2 outside the brain. Similar future studies will permit to unambiguously define the direct impact of MeCP2 on tissue dysfunctions. PMID:26098633

  5. Methyl jasmonate affects morphology, number and activity of endoplasmic reticulum bodies in Raphanus sativus root cells.

    PubMed

    Gotté, Maxime; Ghosh, Rajgourab; Bernard, Sophie; Nguema-Ona, Eric; Vicré-Gibouin, Maïté; Hara-Nishimura, Ikuko; Driouich, Azeddine

    2015-01-01

    The endoplasmic reticulum (ER) bodies are ER-derived structures that are found in Brassicaceae species and thought to play a role in defense. Here, we have investigated the occurrence, distribution and function of ER bodies in root cells of Raphanus sativus using a combination of microscopic and biochemical methods. We have also assessed the response of ER bodies to methyl jasmonate (MeJA), a phytohormone that mediates plant defense against wounding and pathogens. Our results show that (i) ER bodies do occur in different root cell types from the root cap region to the differentiation zone; (ii) they do accumulate a PYK10-like protein similar to the major marker protein of ER bodies that is involved in defense in Arabidopsis thaliana; and (iii) treatment of root cells with MeJA causes a significant increase in the number of ER bodies and the activity of β-glucosidases. More importantly, MeJA was found to induce the formation of very long ER bodies that results from the fusion of small ones, a phenomenon that has not been reported in any other study so far. These findings demonstrate that MeJA impacts the number and morphology of functional ER bodies and stimulates ER body enzyme activities, probably to participate in defense responses of radish root. They also suggest that these structures may provide a defensive system specific to root cells.

  6. Methyl jasmonate affects morphology, number and activity of endoplasmic reticulum bodies in Raphanus sativus root cells.

    PubMed

    Gotté, Maxime; Ghosh, Rajgourab; Bernard, Sophie; Nguema-Ona, Eric; Vicré-Gibouin, Maïté; Hara-Nishimura, Ikuko; Driouich, Azeddine

    2015-01-01

    The endoplasmic reticulum (ER) bodies are ER-derived structures that are found in Brassicaceae species and thought to play a role in defense. Here, we have investigated the occurrence, distribution and function of ER bodies in root cells of Raphanus sativus using a combination of microscopic and biochemical methods. We have also assessed the response of ER bodies to methyl jasmonate (MeJA), a phytohormone that mediates plant defense against wounding and pathogens. Our results show that (i) ER bodies do occur in different root cell types from the root cap region to the differentiation zone; (ii) they do accumulate a PYK10-like protein similar to the major marker protein of ER bodies that is involved in defense in Arabidopsis thaliana; and (iii) treatment of root cells with MeJA causes a significant increase in the number of ER bodies and the activity of β-glucosidases. More importantly, MeJA was found to induce the formation of very long ER bodies that results from the fusion of small ones, a phenomenon that has not been reported in any other study so far. These findings demonstrate that MeJA impacts the number and morphology of functional ER bodies and stimulates ER body enzyme activities, probably to participate in defense responses of radish root. They also suggest that these structures may provide a defensive system specific to root cells. PMID:25305245

  7. Ablation of the mTORC2 component rictor in brain or Purkinje cells affects size and neuron morphology.

    PubMed

    Thomanetz, Venus; Angliker, Nico; Cloëtta, Dimitri; Lustenberger, Regula M; Schweighauser, Manuel; Oliveri, Filippo; Suzuki, Noboru; Rüegg, Markus A

    2013-04-15

    The mammalian target of rapamycin (mTOR) assembles into two distinct multi-protein complexes called mTORC1 and mTORC2. Whereas mTORC1 is known to regulate cell and organismal growth, the role of mTORC2 is less understood. We describe two mouse lines that are devoid of the mTORC2 component rictor in the entire central nervous system or in Purkinje cells. In both lines neurons were smaller and their morphology and function were strongly affected. The phenotypes were accompanied by loss of activation of Akt, PKC, and SGK1 without effects on mTORC1 activity. The striking decrease in the activation and expression of several PKC isoforms, the subsequent loss of activation of GAP-43 and MARCKS, and the established role of PKCs in spinocerebellar ataxia and in shaping the actin cytoskeleton strongly suggest that the morphological deficits observed in rictor-deficient neurons are mediated by PKCs. Together our experiments show that mTORC2 has a particularly important role in the brain and that it affects size, morphology, and function of neurons.

  8. Clinorotation affects soybean seedling morphology

    NASA Technical Reports Server (NTRS)

    Hilaire, Emmanuel; Guikema, James A.; Brown, Christopher S.

    1995-01-01

    Although spaceflight does not appear to significantly affect seed germination, it can influence subsequent plant growth. On STS-3 and SL-2, decreased growth (measured as plant length, fresh weight, and dry weight) was noted for pine, oat, and mung bean. In the CHROMEX-01 and 02 experiments with Haplopappus and in the CHROMEX-03 experiment with Arabidopsis, enhanced root growth was noted in the space-grown plants. In the CHROMEX-04 experiments with wheat, both leaf fresh weight and leaf area were diminished in the space-grown plants but there was no difference in total plant height (CS Brown, HG Levine, and AD Krikorian, unpublished data). These data suggest that microgravity impacts growth by whole plant partitioning of the assimilates. The objective of the present study was to determine the influence of clinorotation on the growth and the morphology of soybean seedlings grown in the Biological Research In Canister (BRIC) flight hardware. This experiment provided baseline data for a spaceflight experiment (BRIC-3) flown on STS-63 (February 3-11, 1995).

  9. Tol-Pal proteins are critical cell envelope components of Erwinia chrysanthemi affecting cell morphology and virulence.

    PubMed

    Dubuisson, Jean-François; Vianney, Anne; Hugouvieux-Cotte-Pattat, Nicole; Lazzaroni, Jean Claude

    2005-10-01

    The tol-pal genes are necessary for maintaining the outer-membrane integrity of Gram-negative bacteria. These genes were first described in Escherichia coli, and more recently in several other species. They are involved in the pathogenesis of E. coli, Haemophilus ducreyi, Vibrio cholerae and Salmonella enterica. The role of the tol-pal genes in bacterial pathogenesis was investigated in the phytopathogenic enterobacterium Erwinia chrysanthemi, assuming that this organism might be a good model for such a study. The whole Er. chrysanthemi tol-pal region was characterized. Tol-Pal proteins, except TolA, showed high identity scores with their E. coli homologues. Er. chrysanthemi mutants were constructed by introducing a uidA-kan cassette in the ybgC, tolQ, tolA, tolB, pal and ybgF genes. All the mutants were hypersensitive to bile salts. Mutations in tolQ, tolA, tolB and pal were deleterious for the bacteria, which required high concentrations of sugars or osmoprotectants for their viability. Consistent with this observation, they were greatly impaired in their cell morphology and division, which was evidenced by observations of cell filaments, spherical forms, membrane blebbing and mislocalized bacterial septa. Moreover, tol-pal mutants showed a reduced virulence in a potato tuber model and on chicory leaves. This could be explained by a combination of impaired phenotypes in the tol-pal mutants, such as reduced growth and motility and a decreased production of pectate lyases, the major virulence factor of Er. chrysanthemi.

  10. Cortisol affects tight junction morphology between pavement cells of rainbow trout gills in single-seeded insert culture.

    PubMed

    Sandbichler, Adolf Michael; Farkas, Julia; Salvenmoser, Willi; Pelster, Bernd

    2011-12-01

    A primary culture system of rainbow trout gill pavement cells grown on permeable support (single-seeded insert, SSI) was used to examine histological and physiological changes induced by the addition of the corticosteroid hormone cortisol. Pavement cell epithelia were cultured under symmetrical conditions (L15 apical/L15 basolateral) and developed a high transepithelial resistance (TER, 6.84 ± 1.99 kΩ cm(2), mean ± SEM) with a low phenol red diffusion rate (PRD, 0.15 ± 0.03 μmol l(-1)/day). Addition of cortisol to the basolateral compartment increased TER twofold and reduced PRD threefold over a 5-day period. A similar increase in TER could be seen after 24 h apical freshwater (FW) in control cultures. In cortisol-treated cultures FW exposure did not change TER, but PRD increased significantly. Histochemical staining of the cytoskeleton of cells in SSI culture revealed a morphological partitioning into a single mucosal layer of polarized, polygonal cells featuring cortical F-actin rings which were comparable to F-actin rings of epithelial cells on the lamellar and filamental surface, and several unorganized serosal layers of cells with F-actin stress fibers. Addition of cortisol increased cell density by 18% and in the mucosal layer it led to smaller, less polygonal cells with increased height and increased cell contact area. In transmission electron microscopic images two pairs of cytoplasmatic electron-dense structures confining the zonula occludens apically and basally toward the zonula adhaerens were found. Addition of cortisol increased the distance between those paired structures, hence led to deeper tight junctions. The cortisol-induced increase in barrier properties, therefore, involves a structural fortification of the tight junctions which was not generally modified by a short 24-h apical freshwater stress. These results identify cortisol as a regulator of tight junction morphology between pavement cells of euryhaline fish such as the

  11. Red blood cell morphology.

    PubMed

    Ford, J

    2013-06-01

    The foundation of laboratory hematologic diagnosis is the complete blood count and review of the peripheral smear. In patients with anemia, the peripheral smear permits interpretation of diagnostically significant red blood cell (RBC) findings. These include assessment of RBC shape, size, color, inclusions, and arrangement. Abnormalities of RBC shape and other RBC features can provide key information in establishing a differential diagnosis. In patients with microcytic anemia, RBC morphology can increase or decrease the diagnostic likelihood of thalassemia. In normocytic anemias, morphology can assist in differentiating among blood loss, marrow failure, and hemolysis-and in hemolysis, RBC findings can suggest specific etiologies. In macrocytic anemias, RBC morphology can help guide the diagnostic considerations to either megaloblastic or nonmegaloblastic causes. Like all laboratory tests, RBC morphologies must be interpreted with caution, particularly in infants and children. When used properly, RBC morphology can be a key tool for laboratory hematology professionals to recommend appropriate clinical and laboratory follow-up and to select the best tests for definitive diagnosis. PMID:23480230

  12. Red blood cell morphology.

    PubMed

    Ford, J

    2013-06-01

    The foundation of laboratory hematologic diagnosis is the complete blood count and review of the peripheral smear. In patients with anemia, the peripheral smear permits interpretation of diagnostically significant red blood cell (RBC) findings. These include assessment of RBC shape, size, color, inclusions, and arrangement. Abnormalities of RBC shape and other RBC features can provide key information in establishing a differential diagnosis. In patients with microcytic anemia, RBC morphology can increase or decrease the diagnostic likelihood of thalassemia. In normocytic anemias, morphology can assist in differentiating among blood loss, marrow failure, and hemolysis-and in hemolysis, RBC findings can suggest specific etiologies. In macrocytic anemias, RBC morphology can help guide the diagnostic considerations to either megaloblastic or nonmegaloblastic causes. Like all laboratory tests, RBC morphologies must be interpreted with caution, particularly in infants and children. When used properly, RBC morphology can be a key tool for laboratory hematology professionals to recommend appropriate clinical and laboratory follow-up and to select the best tests for definitive diagnosis.

  13. Factors affecting spermatozoa morphology in beef bulls

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The objective of this study was to evaluate factors affecting sperm morphology of bulls (n=908) collected at 320 days of age. Bulls were a composite breed (50% Red Angus, 25% Charolais, and 25% Tarentaise) born from 2002 to 2008 to dams fed levels of feed during mid and late gestation that were expe...

  14. Natural polyamines and synthetic analogs modify the growth and the morphology of Pyrus communis pollen tubes affecting ROS levels and causing cell death.

    PubMed

    Aloisi, Iris; Cai, Giampiero; Tumiatti, Vincenzo; Minarini, Anna; Del Duca, Stefano

    2015-10-01

    Polyamines (PAs) are small molecules necessary for pollen maturation and tube growth. Their role is often controversial, since they may act as pro-survival factors as well as factors promoting Programmed Cell Death (PCD). The aim of the present work was to evaluate the effect of exogenous PAs on the apical growth of pear (Pyrus communis) pollen tube and to understand if PAs and reactive oxygen species (ROS) are interconnected in the process of tip-growth. In the present study besides natural PAs, also aryl-substituted spermine and methoctramine (Met 6-8-6) analogs were tested. Among the natural PAs, Spm showed strongest effects on tube growth. Spm entered through the pollen tube tip, then diffused in the sub-apical region that underwent drastic morphological changes, showing enlarged tip. Analogs were mostly less efficient than natural PAs but BD23, an asymmetric synthetic PAs bearing a pyridine ring, showed similar effects. These effects were related to the ability of PAs to cause the decrease of ROS level in the apical zone, leading to cell death, counteracted by the caspase-3 inhibitor Ac-DEVD-CHO (DEVD). In conclusions, ROS are essential for pollen germination and a strict correlation between ROS regulation and PA concentration is reported. Moreover, an imbalance between ROS and PAs can be detrimental thereby driving pollen toward cell death. PMID:26398794

  15. Natural polyamines and synthetic analogs modify the growth and the morphology of Pyrus communis pollen tubes affecting ROS levels and causing cell death.

    PubMed

    Aloisi, Iris; Cai, Giampiero; Tumiatti, Vincenzo; Minarini, Anna; Del Duca, Stefano

    2015-10-01

    Polyamines (PAs) are small molecules necessary for pollen maturation and tube growth. Their role is often controversial, since they may act as pro-survival factors as well as factors promoting Programmed Cell Death (PCD). The aim of the present work was to evaluate the effect of exogenous PAs on the apical growth of pear (Pyrus communis) pollen tube and to understand if PAs and reactive oxygen species (ROS) are interconnected in the process of tip-growth. In the present study besides natural PAs, also aryl-substituted spermine and methoctramine (Met 6-8-6) analogs were tested. Among the natural PAs, Spm showed strongest effects on tube growth. Spm entered through the pollen tube tip, then diffused in the sub-apical region that underwent drastic morphological changes, showing enlarged tip. Analogs were mostly less efficient than natural PAs but BD23, an asymmetric synthetic PAs bearing a pyridine ring, showed similar effects. These effects were related to the ability of PAs to cause the decrease of ROS level in the apical zone, leading to cell death, counteracted by the caspase-3 inhibitor Ac-DEVD-CHO (DEVD). In conclusions, ROS are essential for pollen germination and a strict correlation between ROS regulation and PA concentration is reported. Moreover, an imbalance between ROS and PAs can be detrimental thereby driving pollen toward cell death.

  16. Disruption of the pdhB Pyruvate Dehydrogenase Gene Affects Colony Morphology, In Vitro Growth and Cell Invasiveness of Mycoplasma agalactiae

    PubMed Central

    Hegde, Shivanand; Rosengarten, Renate; Chopra-Dewasthaly, Rohini

    2015-01-01

    The utilization of available substrates, the metabolic potential and the growth rates of bacteria can play significant roles in their pathogenicity. This study concentrates on Mycoplasma agalactiae, which causes significant economic losses through its contribution to contagious agalactia in small ruminants by as yet unknown mechanisms. This lack of knowledge is primarily due to its fastidious growth requirements and the scarcity of genetic tools available for its manipulation and analysis. Transposon mutagenesis of M. agalactiae type strain PG2 resulted in several disruptions throughout the genome. A mutant defective in growth in vitro was found to have a transposon insertion in the pdhB gene, which encodes a component of the pyruvate dehydrogenase complex. This growth difference was quite significant during the actively dividing logarithmic phase but a gradual recovery was observed as the cells approached stationary phase. The mutant also exhibited a different and smaller colony morphology compared to the wild type strain PG2. For complementation, pdhAB was cloned downstream of a strong vpma promoter and upstream of a lacZ reporter gene in a newly constructed complementation vector. When transformed with this vector the pdhB mutant recovered its normal growth and colony morphology. Interestingly, the pdhB mutant also had significantly reduced invasiveness in HeLa cells, as revealed by double immunofluorescence staining. This deficiency was recovered in the complemented strain, which had invasiveness comparable to that of PG2. Taken together, these data indicate that pyruvate dehydrogenase might be an important player in infection with and colonization by M. agalactiae. PMID:25799063

  17. Temperature affects the silicate morphology in a diatom

    PubMed Central

    Javaheri, N.; Dries, R.; Burson, A.; Stal, L. J.; Sloot, P. M. A.; Kaandorp, J. A.

    2015-01-01

    Silica deposition by diatoms, a common component of the phytoplankton, has attracted considerable interest given the importance in ecology and materials science. There has recently been a great deal of research into the biological control of biosilicifcation, yet the in vivo physical and chemical effects have not been quantitatively investigated. We have grown the marine diatom Thalassiosira pseudonana in batch culture at three temperatures (14o, 18o, and 23 °C). We observed three distinct temperature-dependent growth phases. The morphology of silica was investigated using scanning electron microscopy followed by image analysis and supervised learning. The silica in the valves of the same species showed different structures: a mesh-like pattern in silicon-rich cultures and a tree-like pattern in silicon-limited cultures. Moreover, temperature affected this silica pattern, especially in silicon-limited cultures. We conclude that cells grown at 14 °C and 18 °C divide more successfully in Si-limited conditions by developing a tree-like pattern (lower silicification). PMID:26113515

  18. Cell morphology in injectable nanostructured biosynthetic hydrogels.

    PubMed

    Yom-Tov, Ortal; Seliktar, Dror; Bianco-Peled, Havazelet

    2014-12-01

    Even though inducing structural features on the nanometric scale has been shown to be a powerful tool in tissue engineering, almost all nanostructuring techniques available today cannot be applied to injectable hydrogel scaffolds. The current research explores such a novel technique and its effect on scaffold's properties, cell morphology, and cell-material interaction. Nanostructuring is achieved by covalently binding Pluronic(®) F127 molecules to biosynthetic hydrogels. Analysis of cell morphology revealed spindled cell morphologies at day 4 in culture. The bound Pluronic(®) F127 diminished the swelling ability and enhanced the Young modulus, thus indicating that the bound molecules crosslink the hydrogel. The relation between matrix characteristics and cell morphology was analyzed and the importance of nanostructuring was demonstrated.

  19. Reelin signaling directly affects radial glia morphology and biochemical maturation.

    PubMed

    Hartfuss, Eva; Förster, Eckart; Bock, Hans H; Hack, Michael A; Leprince, Pierre; Luque, Juan M; Herz, Joachim; Frotscher, Michael; Götz, Magdalena

    2003-10-01

    Radial glial cells are characterized, besides their astroglial properties, by long radial processes extending from the ventricular zone to the pial surface, a crucial feature for the radial migration of neurons. The molecular signals that regulate this characteristic morphology, however, are largely unknown. We show an important role of the secreted molecule reelin for the establishment of radial glia processes. We describe a significant reduction in ventricular zone cells with long radial processes in the absence of reelin in the cortex of reeler mutant mice. These defects were correlated to a decrease in the content of brain lipid-binding protein (Blbp) and were detected exclusively in the cerebral cortex, but not in the basal ganglia of reeler mice. Conversely, reelin addition in vitro increased the Blbp content and process extension of radial glia from the cortex, but not the basal ganglia. Isolation of radial glia by fluorescent-activated cell sorting showed that these effects are due to direct signaling of reelin to radial glial cells. We could further demonstrate that this signaling requires Dab1, as the increase in Blbp upon reelin addition failed to occur in Dab1-/- mice. Taken together, these results unravel a novel role of reelin signaling to radial glial cells that is crucial for the regulation of their Blbp content and characteristic morphology in a region-specific manner.

  20. Effect of hydroxyapatite surface morphology on cell adhesion.

    PubMed

    Iwamoto, Takashi; Hieda, Yohki; Kogai, Yasumichi

    2016-12-01

    We obtained hydroxyapatite (HAp) materials as a block by mixing HAp nanoparticles and polymer, and then calcining the mixtures. The surface morphology of the HAp materials was tuned by varying heat treatment conditions. After calcining the mixtures at 1200 or 800°C for 4h, the surface morphology of the HAp materials was flat or convexo-concave, respectively. The flat surface morphology, which showed micrometer-ordered grain boundaries, was formed by the aggregation of HAp nanoparticles. On the other hand, the convexo-concave surface morphology resulted from the agglomeration of HAp nanoparticles after heat treatment at 800°C for 4h with nanometer-ordered particle size. We tested cell adhesion to HAp materials with flat or convexo-concave surface morphology and found that cells adhered well to the flat HAp materials but not to the convexo-concave HAp materials. This technique for selectively preparing HAp materials with flat or convexo-concave surface morphology was very easy because we merely mixed commercial HAp nanoparticles with polymer and then calcined the mixtures. As a result, the heat treatment temperature affected the surface morphology of our HAp materials, and their surface morphologies contributed to cell adhesion independently of other material properties. PMID:27612825

  1. Effect of hydroxyapatite surface morphology on cell adhesion.

    PubMed

    Iwamoto, Takashi; Hieda, Yohki; Kogai, Yasumichi

    2016-12-01

    We obtained hydroxyapatite (HAp) materials as a block by mixing HAp nanoparticles and polymer, and then calcining the mixtures. The surface morphology of the HAp materials was tuned by varying heat treatment conditions. After calcining the mixtures at 1200 or 800°C for 4h, the surface morphology of the HAp materials was flat or convexo-concave, respectively. The flat surface morphology, which showed micrometer-ordered grain boundaries, was formed by the aggregation of HAp nanoparticles. On the other hand, the convexo-concave surface morphology resulted from the agglomeration of HAp nanoparticles after heat treatment at 800°C for 4h with nanometer-ordered particle size. We tested cell adhesion to HAp materials with flat or convexo-concave surface morphology and found that cells adhered well to the flat HAp materials but not to the convexo-concave HAp materials. This technique for selectively preparing HAp materials with flat or convexo-concave surface morphology was very easy because we merely mixed commercial HAp nanoparticles with polymer and then calcined the mixtures. As a result, the heat treatment temperature affected the surface morphology of our HAp materials, and their surface morphologies contributed to cell adhesion independently of other material properties.

  2. How molecular interactions affect crystal morphology: the case of haloperidol.

    PubMed

    Li Destri, Giovanni; Marrazzo, Agostino; Rescifina, Antonio; Punzo, Francesco

    2011-11-01

    The tableting behaviour of drugs can be dramatically affected by changes in the crystal habit of the drug molecule. Pharmaceutical companies are therefore interested in the morphology prediction as a possible tool to optimise the industrial process. Molecular mechanics calculations embedded in dedicated software together with X-ray diffraction analysis were used to enlighten the structural properties of 4-[4-(4-chlorophenyl)-4-hydroxypiperidin-1-yl]-1-(4-fluorophenyl)butan-1-one-whose commercial name is haloperidol--an antipsychotic drug that contributed to the progress and revolution of psychiatric care. We defined, by means of X-ray powder diffraction, which--or how much--of the two crystallographic structures present in the Cambridge Crystallographic Database represents the commercial crystalline powder. Once the correct structure was selected, the whole structural analysis was carried out as a comparison with the already deposited structures. The available single crystal structure was used to model the X-ray powder diffraction pattern. The "real" structure was then optimised by means of molecular mechanics and the crystal morphology of the compounds was predicted with different computational methods. Analogies and differences among the different morphologies, together with the potential role of several solvents were used to try to bridge the gap between the molecular structure--that is, the atomic point of view--and the crystal habit. PMID:21656519

  3. Personality and morphological traits affect pigeon survival from raptor attacks

    PubMed Central

    Santos, Carlos D.; Cramer, Julia F.; Pârâu, Liviu G.; Miranda, Ana C.; Wikelski, Martin; Dechmann, Dina K. N.

    2015-01-01

    Personality traits have recently been shown to impact fitness in different animal species, potentially making them similarly relevant drivers as morphological and life history traits along the evolutionary pathways of organisms. Predation is a major force of natural selection through its deterministic effects on individual survival, but how predation pressure has helped to shape personality trait selection, especially in free-ranging animals, remains poorly understood. We used high-precision GPS tracking to follow whole flocks of homing pigeons (Columba livia) with known personalities and morphology during homing flights where they were severely predated by raptors. This allowed us to determine how the personality and morphology traits of pigeons may affect their risk of being predated by raptors. Our survival model showed that individual pigeons, which were more tolerant to human approach, slower to escape from a confined environment, more resistant to human handling, with larger tarsi, and with lighter plumage, were more likely to be predated by raptors. We provide rare empirical evidence that the personality of prey influences their risk of being predated under free-ranging circumstances. PMID:26489437

  4. Personality and morphological traits affect pigeon survival from raptor attacks.

    PubMed

    Santos, Carlos D; Cramer, Julia F; Pârâu, Liviu G; Miranda, Ana C; Wikelski, Martin; Dechmann, Dina K N

    2015-10-22

    Personality traits have recently been shown to impact fitness in different animal species, potentially making them similarly relevant drivers as morphological and life history traits along the evolutionary pathways of organisms. Predation is a major force of natural selection through its deterministic effects on individual survival, but how predation pressure has helped to shape personality trait selection, especially in free-ranging animals, remains poorly understood. We used high-precision GPS tracking to follow whole flocks of homing pigeons (Columba livia) with known personalities and morphology during homing flights where they were severely predated by raptors. This allowed us to determine how the personality and morphology traits of pigeons may affect their risk of being predated by raptors. Our survival model showed that individual pigeons, which were more tolerant to human approach, slower to escape from a confined environment, more resistant to human handling, with larger tarsi, and with lighter plumage, were more likely to be predated by raptors. We provide rare empirical evidence that the personality of prey influences their risk of being predated under free-ranging circumstances.

  5. Factors affecting the morphology of benzoyl peroxide microsponges.

    PubMed

    Nokhodchi, Ali; Jelvehgari, Mitra; Siahi, M Reza; Mozafari, M Reza

    2007-01-01

    Benzoyl peroxide (BPO) is primarily used in the treatment of mild to moderate acne. However, its application is associated with skin irritation. It has been shown that encapsulation and controlled release of BPO could reduce the side effect while also reducing percutaneous absorption when administered to the skin. The aim of the present investigation was to design and formulate an appropriate encapsulated form of BPO, using microsponge technology, and explore the parameters affecting the morphology and other characteristics of the resultant products employing scanning electron microscopy (SEM). Benzoyl peroxide particles were prepared using an emulsion solvent diffusion method by adding an organic internal phase containing benzoyl peroxide, ethyl cellulose and dichloromethane into a stirred aqueous phase containing polyvinyl alcohol (PVA). Different concentrations of BPO microsponges were incorporated in lotion formulations and the drug release from these formulations were studied. The SEM micrographs of the BPO microsponges enabled measurement of their size and showed that they were spherical and porous. Results showed that the morphology and particle size of microsponges were affected by drug:polymer ratio, stirring rate and the amount of emulsifier used. The results obtained also showed that an increase in the ratio of drug:polymer resulted in a reduction in the release rate of BPO from the microsponges. The release data showed that the highest and the lowest release rates were obtained from lotions containing plain BPO particles and BPO microsponges with the drug:polymer ratio of 13:1, respectively. The kinetics of release study showed that the release data followed Peppas model and the main mechanism of drug release from BPO microsponges was diffusion.

  6. Twenty Four-Hour Exposure to a 0.12 THz Electromagnetic Field Does Not Affect the Genotoxicity, Morphological Changes, or Expression of Heat Shock Protein in HCE-T Cells.

    PubMed

    Koyama, Shin; Narita, Eijiro; Shimizu, Yoko; Shiina, Takeo; Taki, Masao; Shinohara, Naoki; Miyakoshi, Junji

    2016-01-01

    To investigate the cellular effects of terahertz (THz) exposure, human corneal epithelial (HCE-T) cells derived from human eye were exposed to 0.12 THz radiation at 5 mW/cm² for 24 h, then the genotoxicity, morphological changes, and heat shock protein (Hsp) expression of the cells were examined. There was no statistically significant increase in the micronucleus (MN) frequency of cells exposed to 0.12 THz radiation compared with sham-exposed controls and incubator controls, whereas the MN frequency of cells treated with bleomycin for 1 h (positive control) did increase significantly. Similarly, there were no significant morphological changes in cells exposed to 0.12 THz radiation compared to sham-exposed controls and incubator controls, and Hsp expression (Hsp27, Hsp70, and Hsp90α) was also not significantly different between the three treatments. These results indicate that exposure to 0.12 THz radiation using the present conditions appears to have no or very little effect on MN formation, morphological changes, and Hsp expression in cells derived from human eye. PMID:27527204

  7. Twenty Four-Hour Exposure to a 0.12 THz Electromagnetic Field Does Not Affect the Genotoxicity, Morphological Changes, or Expression of Heat Shock Protein in HCE-T Cells

    PubMed Central

    Koyama, Shin; Narita, Eijiro; Shimizu, Yoko; Shiina, Takeo; Taki, Masao; Shinohara, Naoki; Miyakoshi, Junji

    2016-01-01

    To investigate the cellular effects of terahertz (THz) exposure, human corneal epithelial (HCE-T) cells derived from human eye were exposed to 0.12 THz radiation at 5 mW/cm2 for 24 h, then the genotoxicity, morphological changes, and heat shock protein (Hsp) expression of the cells were examined. There was no statistically significant increase in the micronucleus (MN) frequency of cells exposed to 0.12 THz radiation compared with sham-exposed controls and incubator controls, whereas the MN frequency of cells treated with bleomycin for 1 h (positive control) did increase significantly. Similarly, there were no significant morphological changes in cells exposed to 0.12 THz radiation compared to sham-exposed controls and incubator controls, and Hsp expression (Hsp27, Hsp70, and Hsp90α) was also not significantly different between the three treatments. These results indicate that exposure to 0.12 THz radiation using the present conditions appears to have no or very little effect on MN formation, morphological changes, and Hsp expression in cells derived from human eye. PMID:27527204

  8. Twenty Four-Hour Exposure to a 0.12 THz Electromagnetic Field Does Not Affect the Genotoxicity, Morphological Changes, or Expression of Heat Shock Protein in HCE-T Cells.

    PubMed

    Koyama, Shin; Narita, Eijiro; Shimizu, Yoko; Shiina, Takeo; Taki, Masao; Shinohara, Naoki; Miyakoshi, Junji

    2016-08-05

    To investigate the cellular effects of terahertz (THz) exposure, human corneal epithelial (HCE-T) cells derived from human eye were exposed to 0.12 THz radiation at 5 mW/cm² for 24 h, then the genotoxicity, morphological changes, and heat shock protein (Hsp) expression of the cells were examined. There was no statistically significant increase in the micronucleus (MN) frequency of cells exposed to 0.12 THz radiation compared with sham-exposed controls and incubator controls, whereas the MN frequency of cells treated with bleomycin for 1 h (positive control) did increase significantly. Similarly, there were no significant morphological changes in cells exposed to 0.12 THz radiation compared to sham-exposed controls and incubator controls, and Hsp expression (Hsp27, Hsp70, and Hsp90α) was also not significantly different between the three treatments. These results indicate that exposure to 0.12 THz radiation using the present conditions appears to have no or very little effect on MN formation, morphological changes, and Hsp expression in cells derived from human eye.

  9. Gravitational environment produced by a superconducting magnet affects osteoblast morphology and functions

    NASA Astrophysics Data System (ADS)

    Qian, Airong; Zhang, Wei; Weng, Yuanyuan; Tian, Zongcheng; Di, Shengmeng; Yang, Pengfei; Yin, Dachuan; Hu, Lifang; Wang, Zhe; Xu, Huiyun; Shang, Peng

    The aims of this study are to investigate the effects of gravitational environment produced by a superconducting magnet on osteoblast morphology, proliferation and adhesion. A superconducting magnet which can produce large gradient high magnetic field (LGHMF) and provide three apparent gravity levels (0g,1gand2g) was employed to simulate space gravity environment. The effects of LGHMF on osteoblast morphology, proliferation, adhesion and the gene expression of fibronectin and collagen I were detected by scanning electron microscopy, immunocytochemistry, adhesion assays and real time PCR, respectively, after exposure of osteoblasts to LGHMF for 24 h. Osteoblast morphology was affected by LGHMF (0g,1gand2g) and the most evident morphology alteration was observed at 0g condition. Proliferative abilities of MC3T3 and MG-63 cell were affected under LGHMF (0g,1gand2g) conditions compared to control condition. The adhesive abilities of MC3T3 and MG-63 cells to extracellular matrix (ECM) proteins (fibronectin, laminin, collagen IV) were also affected by LGHMF (0g,1gand2g), moreover, the effects of LGHMF on osteoblast adhesion to different ECM proteins were different. Fibronectin gene expression in MG63 cells under zero gravity condition was increased significantly compared to other conditions. Collagen I gene expression in MG-63 and MC3T3 cells was altered by both magnetic field and alerted gravity. The study indicates that the superconducting magnet which can produce LGHMF may be a novel ground-based space gravity simulator and can be used for biological experiment at cellular level.

  10. Plastic solar cell interface and morphological characterization

    NASA Astrophysics Data System (ADS)

    Guralnick, Brett W.

    Plastic solar cell research has become an intense field of study considering these devices may be lightweight, flexible and reduce the cost of photovoltaic devices. The active layer of plastic solar cells are a combination of two organic components which blend to form an internal morphology. Due to the poor electrical transport properties of the organic components it is important to understand how the morphology forms in order to engineer these materials for increased efficiency. The focus of this thesis is a detailed study of the interfaces between the plastic solar cell layers and the morphology of the active layer. The system studied in detail is a blend of P3HT and PCBM that acts as the primary absorber, which is the electron donor, and the electron acceptor, respectively. The key morphological findings are, while thermal annealing increases the crystallinity parallel to the substrate, the morphology is largely unchanged following annealing. The deposition and mixing conditions of the bulk heterojunction from solution control the starting morphology. The spin coating speed, concentration, solvent type, and solution mixing time are all critical variables in the formation of the bulk heterojunction. In addition, including the terminals or inorganic layers in the analysis is critical because the inorganic surface properties influence the morphology. Charge transfer in the device occurs at the material interfaces, and a highly resistive transparent conducting oxide layer limits device performance. It was discovered that the electron blocking layer between the transparent conducting oxide and the bulk heterojunction is compromised following annealing. The electron acceptor material can diffuse into this layer, a location which does not benefit device performance. Additionally, the back contact deposition is important since the organic material can be damaged by the thermal evaporation of Aluminum, typically used for plastic solar cells. Depositing a thin thermal and

  11. Morphological classification of plant cell deaths

    PubMed Central

    van Doorn, W G; Beers, E P; Dangl, J L; Franklin-Tong, V E; Gallois, P; Hara-Nishimura, I; Jones, A M; Kawai-Yamada, M; Lam, E; Mundy, J; Mur, L A J; Petersen, M; Smertenko, A; Taliansky, M; Van Breusegem, F; Wolpert, T; Woltering, E; Zhivotovsky, B; Bozhkov, P V

    2011-01-01

    Programmed cell death (PCD) is an integral part of plant development and of responses to abiotic stress or pathogens. Although the morphology of plant PCD is, in some cases, well characterised and molecular mechanisms controlling plant PCD are beginning to emerge, there is still confusion about the classification of PCD in plants. Here we suggest a classification based on morphological criteria. According to this classification, the use of the term ‘apoptosis' is not justified in plants, but at least two classes of PCD can be distinguished: vacuolar cell death and necrosis. During vacuolar cell death, the cell contents are removed by a combination of autophagy-like process and release of hydrolases from collapsed lytic vacuoles. Necrosis is characterised by early rupture of the plasma membrane, shrinkage of the protoplast and absence of vacuolar cell death features. Vacuolar cell death is common during tissue and organ formation and elimination, whereas necrosis is typically found under abiotic stress. Some examples of plant PCD cannot be ascribed to either major class and are therefore classified as separate modalities. These are PCD associated with the hypersensitive response to biotrophic pathogens, which can express features of both necrosis and vacuolar cell death, PCD in starchy cereal endosperm and during self-incompatibility. The present classification is not static, but will be subject to further revision, especially when specific biochemical pathways are better defined. PMID:21494263

  12. Environmental properties set cell mechanics and morphology

    NASA Astrophysics Data System (ADS)

    Janmey, Paul

    2012-02-01

    Many cell types are sensitive to mechanical signals that are produced either by application of exogenous force to their surfaces, or by the resistance that their surroundings place on forces generated by the cells themselves. Cell morphology, motility, proliferation, and protein expression all change in response to substrate stiffness. Changing the elastic moduli of substrates alters the formation of focal adhesions, the assembly of actin filaments into bundles, and the stability of intermediate filaments. The range of stiffness over which different primary cell types respond can vary over a wide range and generally reflects the elastic modulus of the tissue from which these cells were isolated. Mechanosensing depends on the type of adhesion receptor by which the cell binds, and therefore on both the molecular composition of the extracellular matrix and the nature of its link to the cytoskeleton. Many cell types can alter their own stiffness to match that of the substrate to which they adhere. The maximal elastic modulus that cells such as fibroblasts can attain is similar to that of crosslinked actin networks at the concentrations in the cell cortex. The precise mechanisms of mechanosensing are not well defined, but they presumably require an elastic connection between cell and substrate, mediated by transmembrane proteins. The viscoelastic properties of different extracellular matrices and cytoskeletal elements strongly influence the response of cells to mechanical signals, and the unusual non-linear elasticity of many biopolymer gels, characterized by strain-stiffening, leads to novel mechanisms by which cells alter their stiffness by engagement of molecular motors that produce internal stresses. Cell cortical elasticity is dominated by cytoskeletal polymer networks and can be modulated by internal tension. Simultaneous control of substrate stiffness and adhesive patterns suggests that stiffness sensing occurs on a length scale much larger than single molecular

  13. Glioblastoma with signet ring cell morphology: A diagnostic challenge

    PubMed Central

    Krishnamoorthy, Naveen; Veldore, Vidya; Sridhar, P. S.; Govindrajan, M. J.; Prabhudesai, Shilpa; Hazarika, Digantha; Ajaikumar, B. S.

    2016-01-01

    Glioblastoma (WHO Grade IV), the most frequent malignant brain tumor, can have varied morphologic variations like epithelial/glandular structures, granular cells, and lipidized cells. Glioblastoma with signet ring cell morphology is very unusual and can mimic a metastatic carcinoma. These rare tumors may be just a morphological variant or may signify a different carcinogenic pathway. PMID:27366281

  14. Factors Affecting the Acquisition of Plural Morphology in Jordanian Arabic

    ERIC Educational Resources Information Center

    Albirini, Abdulkafi

    2015-01-01

    This study investigates the development of plural morphology in Jordanian Arab children, and explores the role of the predictability, transparency, productivity, and frequency of different plural forms in determining the trajectory that children follow in acquiring this complex inflectional system. The study also re-examines the development of the…

  15. How Word Frequency Affects Morphological Processing in Monolinguals and Bilinguals

    ERIC Educational Resources Information Center

    Lehtonen, Minna; Laine, Matti

    2003-01-01

    The present study investigated processing of morphologically complex words in three different frequency ranges in monolingual Finnish speakers and Finnish-Swedish bilinguals. By employing a visual lexical decision task, we found a differential pattern of results in monolinguals vs. bilinguals. Monolingual Finns seemed to process low frequency and…

  16. Photoperiod affects the diurnal rhythm of hippocampal neuronal morphology of Siberian hamsters.

    PubMed

    Ikeno, Tomoko; Weil, Zachary M; Nelson, Randy J

    2013-11-01

    Individuals of many species can regulate their physiology, morphology, and behavior in response to annual changes of day length (photoperiod). In mammals, the photoperiodic signal is mediated by a change in the duration of melatonin, leading to alterations in gene expressions, neuronal circuits, and hormonal secretion. The hippocampus is one of the most plastic structures in the adult brain and hippocampal neuronal morphology displays photoperiod-induced differences. Because the hippocampus is important for emotional and cognitive behaviors, photoperiod-driven remodeling of hippocampal neurons is implicated in seasonal differences of affect, including seasonal affective disorder (SAD) in humans. Because neuronal architecture is also affected by the day-night cycle in several brain areas, we hypothesized that hippocampal neuronal morphology would display a diurnal rhythm and that day length would influence that rhythm. In the present study, we examined diurnal and seasonal differences in hippocampal neuronal morphology, as well as mRNA expression of the neurotrophic factors (i.e., brain-derived neurotrophic factor [Bdnf], tropomyosin receptor kinase B [trkB; a receptor for BDNF], and vascular endothelial growth factor [Vegf]) and a circadian clock gene, Bmal1, in the hippocampus of Siberian hamsters. Diurnal rhythms in total length of dendrites, the number of primary dendrites, dendritic complexity, and distance of the furthest intersection from the cell body were observed only in long-day animals; however, diurnal rhythms in the number of branch points and mean length of segments were observed only in short-day animals. Spine density of dendrites displayed diurnal rhythmicity with different peak times between the CA1 and DG subregions and between long and short days. These results indicate that photoperiod affects daily morphological changes of hippocampal neurons and the daily rhythm of spine density, suggesting the possibility that photoperiod-induced adjustments

  17. The role of anisotropy in cell morphology

    NASA Astrophysics Data System (ADS)

    Schakenraad, Koen; Pomp, Wim; Merks, Roeland; Schmidt, Thomas; Giomi, Luca

    The shape of adhering cells is determined by the interplay between contractile forces, arising from the cytoskeleton, and the resistance of the underlying substrate. In particular, experiments with fibroblasts on an elastic micro-pillar array show that fibroblasts posess a high degree of orientational order of the actin stress fibers. This anisotropy causes the shape of the cell edge to deviate from the shape of cells with an isotropic cytoskeleton. We present a model that describes the contractility of the cytoskeleton as a combination of directed forces, in the direction of stress fibers, and isotropic forces. We found that cell morphology is described by an anisotropic generalization of the Young-Laplace law, which describes the cell edges as parts of an ellipse. Experiments on the shape of and adhesion forces on fibroblasts show good agreement with our model. Our work highlights the strong coupling between the organization of the internal cytoskeleton and the shapes and forces on the outside of the cell.

  18. Morphology Studies of Polymer Bulk Heterojunction Solar Cells

    NASA Astrophysics Data System (ADS)

    Moon, Ji Sun

    Energy is a prerequisite for creating and sustaining life. The need for energy increases globally as the world's population and economy grow. However, conventional energy sources---fossil fuels---generate carbon dioxide and contribute to global warming, perhaps the most serious environmental problem of our time. Carbon dioxide-free energy is required to stop global warming. Polymer solar cells have been attracting a great deal of interest as a source of renewable energy with a great potential for low cost. Polymer bulk heterojunction (BHJ) solar cells have been greatly improved; the power conversion efficiency is already up to 9.2% making the future of the polymer solar cell very promising. This thesis is a study of the morphology of polymer:fullerene BHJ, one of the most critical and challenging parts of high efficiency polymer solar cells. To discover the morphology, cross-section as well as top-down transmission electron microscopy were used. The contrast was achieved by utilizing phase contrast microscopy. Thermal annealing, dependence of BHJ thickness, processing additives, solution sequential process and solution sequential process with the use of cosolvent that affects/controls the BHJ morphology are studied in detail.

  19. Clinorotation affects morphology and ethylene production in soybean seedlings

    NASA Technical Reports Server (NTRS)

    Hilaire, E.; Peterson, B. V.; Guikema, J. A.; Brown, C. S.; Sager, J. C. (Principal Investigator)

    1996-01-01

    The microgravity environment of spaceflight influences growth, morphology and metabolism in etiolated germinating soybean. To determine if clinorotation will similarly impact these processes, we conducted ground-based studies in conjunction with two space experiment opportunities. Soybean (Glycine max [L.] Merr.) seeds were planted within BRIC (Biological Research In Canister) canisters and grown for seven days at 20 degrees C under clinorotation (1 rpm) conditions or in a stationary upright mode. Gas samples were taken daily and plants were harvested after seven days for measurement of growth and morphology. Compared to the stationary upright controls, plants exposed to clinorotation exhibited increased root length (125% greater) and fresh weight (42% greater), whereas shoot length and fresh weight decreased by 33% and 16% respectively. Plants grown under clinorotation produced twice as much ethylene as the stationary controls. Seedlings treated with triiodo benzoic acid (TIBA), an auxin transport inhibitor, under clinorotation produced 50% less ethylene than the untreated control subjected to the same gravity treatment, whereas a treatment with 2,4-D increased ethylene by five-fold in the clinorotated plants. These data suggest that slow clinorotation influences biomass partitioning and ethylene production in etiolated soybean plants.

  20. Habitat use affects morphological diversification in dragon lizards

    PubMed Central

    COLLAR, D C; SCHULTE, J A; O’MEARA, B C; LOSOS, J B

    2010-01-01

    Habitat use may lead to variation in diversity among evolutionary lineages because habitats differ in the variety of ways they allow for species to make a living. Here, we show that structural habitats contribute to differential diversification of limb and body form in dragon lizards (Agamidae). Based on phylogenetic analysis and ancestral state reconstructions for 90 species, we find that multiple lineages have independently adopted each of four habitat use types: rock-dwelling, terrestriality, semi-arboreality and arboreality. Given these reconstructions, we fit models of evolution to species’ morphological trait values and find that rock-dwelling and arboreality limit diversification relative to terrestriality and semi-arboreality. Models preferred by Akaike information criterion infer slower rates of size and shape evolution in lineages inferred to occupy rocks and trees, and model-averaged rate estimates are slowest for these habitat types. These results suggest that ground-dwelling facilitates ecomorphological differentiation and that use of trees or rocks impedes diversification. PMID:20345808

  1. How Morphological Constraints Affect Axonal Polarity in Mouse Neurons

    PubMed Central

    Bugnicourt, Ghislain; Saoudi, Yasmina; Andrieux, Annie; Gory-Fauré, Sylvie; Villard, Catherine

    2012-01-01

    Neuronal differentiation is under the tight control of both biochemical and physical information arising from neighboring cells and micro-environment. Here we wished to assay how external geometrical constraints applied to the cell body and/or the neurites of hippocampal neurons may modulate axonal polarization in vitro. Through the use of a panel of non-specific poly-L-lysine micropatterns, we manipulated the neuronal shape. By applying geometrical constraints on the cell body we provided evidence that centrosome location was not predictive of axonal polarization but rather follows axonal fate. When the geometrical constraints were applied to the neurites trajectories we demonstrated that axonal specification was inhibited by curved lines. Altogether these results indicated that intrinsic mechanical tensions occur during neuritic growth and that maximal tension was developed by the axon and expressed on straight trajectories. The strong inhibitory effect of curved lines on axon specification was further demonstrated by their ability to prevent formation of multiple axons normally induced by cytochalasin or taxol treatments. Finally we provided evidence that microtubules were involved in the tension-mediated axonal polarization, acting as curvature sensors during neuronal differentiation. Thus, biomechanics coupled to physical constraints might be the first level of regulation during neuronal development, primary to biochemical and guidance regulations. PMID:22457779

  2. Insights into Embryo Defenses of the Invasive Apple Snail Pomacea canaliculata: Egg Mass Ingestion Affects Rat Intestine Morphology and Growth

    PubMed Central

    Gimeno, Eduardo J.; Heras, Horacio

    2014-01-01

    Background The spread of the invasive snail Pomacea canaliculata is expanding the rat lungworm disease beyond its native range. Their toxic eggs have virtually no predators and unusual defenses including a neurotoxic lectin and a proteinase inhibitor, presumably advertised by a warning coloration. We explored the effect of egg perivitellin fluid (PVF) ingestion on the rat small intestine morphology and physiology. Methodology/Principal Findings Through a combination of biochemical, histochemical, histopathological, scanning electron microscopy, cell culture and feeding experiments, we analyzed intestinal morphology, growth rate, hemaglutinating activity, cytotoxicity and cell proliferation after oral administration of PVF to rats. PVF adversely affects small intestine metabolism and morphology and consequently the standard growth rate, presumably by lectin-like proteins, as suggested by PVF hemaglutinating activity and its cytotoxic effect on Caco-2 cell culture. Short-term effects of ingested PVF were studied in growing rats. PVF-supplemented diet induced the appearance of shorter and wider villi as well as fused villi. This was associated with changes in glycoconjugate expression, increased cell proliferation at crypt base, and hypertrophic mucosal growth. This resulted in a decreased absorptive surface after 3 days of treatment and a diminished rat growth rate that reverted to normal after the fourth day of treatment. Longer exposure to PVF induced a time-dependent lengthening of the small intestine while switching to a control diet restored intestine length and morphology after 4 days. Conclusions/Significance Ingestion of PVF rapidly limits the ability of potential predators to absorb nutrients by inducing large, reversible changes in intestinal morphology and growth rate. The occurrence of toxins that affect intestinal morphology and absorption is a strategy against predation not recognized among animals before. Remarkably, this defense is rather similar to

  3. Double-Staining Method for Differentiation of Morphological Changes and Membrane Integrity of Campylobacter coli Cells

    PubMed Central

    Alonso, Jose L.; Mascellaro, Salvatore; Moreno, Yolanda; Ferrús, María A.; Hernández, Javier

    2002-01-01

    We developed a double-staining procedure involving NanoOrange dye (Molecular Probes, Eugene, Oreg.) and membrane integrity stains (LIVE/DEAD BacLight kit; Molecular Probes) to show the morphological and membrane integrity changes of Campylobacter coli cells during growth. The conversion from a spiral to a coccoid morphology via intermediary forms and the membrane integrity changes of the C. coli cells can be detected with the double-staining procedure. Our data indicate that young or actively growing cells are mainly spiral shaped (green-stained cells), but older cells undergo a degenerative change to coccoid forms (red-stained cells). Club-shaped transition cell forms were observed with NanoOrange stain. Chlorinated drinking water affected the viability but not the morphology of C. coli cells. PMID:12324366

  4. Double-staining method for differentiation of morphological changes and membrane integrity of Campylobacter coli cells.

    PubMed

    Alonso, Jose L; Mascellaro, Salvatore; Moreno, Yolanda; Ferrús, María A; Hernández, Javier

    2002-10-01

    We developed a double-staining procedure involving NanoOrange dye (Molecular Probes, Eugene, Oreg.) and membrane integrity stains (LIVE/DEAD BacLight kit; Molecular Probes) to show the morphological and membrane integrity changes of Campylobacter coli cells during growth. The conversion from a spiral to a coccoid morphology via intermediary forms and the membrane integrity changes of the C. coli cells can be detected with the double-staining procedure. Our data indicate that young or actively growing cells are mainly spiral shaped (green-stained cells), but older cells undergo a degenerative change to coccoid forms (red-stained cells). Club-shaped transition cell forms were observed with NanoOrange stain. Chlorinated drinking water affected the viability but not the morphology of C. coli cells.

  5. Neither respiration nor cytochrome c oxidase affects mitochondrial morphology in Saccharomyces cerevisiae.

    PubMed

    Church, C; Poyton, R O

    1998-06-01

    Previous studies have reported that mitochondrial morphology and volume in yeast cells are linked to cellular respiratory capacity. These studies revealed that mitochondrial morphology in glucose-repressed or anaerobically grown cells, which lack or have reduced levels of respiration, is different from that in fully respiring cells. Although both oxygen deprivation and glucose repression decrease the levels of respiratory chain proteins, they decrease the expression of many non-mitochondrial proteins as well, making it difficult to determine whether it is a defect in respiration or something else that effects mitochondrial morphology. To determine whether mitochondrial morphology is dependent on respiration per se, we used a strain with a null mutation in PET100, a nuclear gene that is specifically required for the assembly of cytochrome c oxidase. Although this strain lacks respiration, the mitochondrial morphology and volumes are both comparable to those found in its respiration-proficient parent. These findings indicate that respiration is not involved in the establishment or maintenance of yeast mitochondrial morphology, and that the previously observed effects of oxygen availability and glucose repression on mitochondrial morphology are not exerted through the respiratory chain. By applying the principle of symmorphosis to these findings, we conclude that the shape and size of the mitochondrial reticulum found in respiring yeast cells is maintained for reasons other than respiration.

  6. Effect of Yeast Cell Morphology, Cell Wall Physical Structure and Chemical Composition on Patulin Adsorption.

    PubMed

    Luo, Ying; Wang, Jianguo; Liu, Bin; Wang, Zhouli; Yuan, Yahong; Yue, Tianli

    2015-01-01

    The capability of yeast to adsorb patulin in fruit juice can aid in substantially reducing the patulin toxic effect on human health. This study aimed to investigate the capability of yeast cell morphology and cell wall internal structure and composition to adsorb patulin. To compare different yeast cell morphologies, cell wall internal structure and composition, scanning electron microscope, transmission electron microscope and ion chromatography were used. The results indicated that patulin adsorption capability of yeast was influenced by cell surface areas, volume, and cell wall thickness, as well as 1,3-β-glucan content. Among these factors, cell wall thickness and 1,3-β-glucan content serve significant functions. The investigation revealed that patulin adsorption capability was mainly affected by the three-dimensional network structure of the cell wall composed of 1,3-β-glucan. Finally, patulin adsorption in commercial kiwi fruit juice was investigated, and the results indicated that yeast cells could adsorb patulin from commercial kiwi fruit juice efficiently. This study can potentially simulate in vitro cell walls to enhance patulin adsorption capability and successfully apply to fruit juice industry. PMID:26295574

  7. Effect of Yeast Cell Morphology, Cell Wall Physical Structure and Chemical Composition on Patulin Adsorption

    PubMed Central

    Luo, Ying; Wang, Jianguo; Liu, Bin; Wang, Zhouli; Yuan, Yahong; Yue, Tianli

    2015-01-01

    The capability of yeast to adsorb patulin in fruit juice can aid in substantially reducing the patulin toxic effect on human health. This study aimed to investigate the capability of yeast cell morphology and cell wall internal structure and composition to adsorb patulin. To compare different yeast cell morphologies, cell wall internal structure and composition, scanning electron microscope, transmission electron microscope and ion chromatography were used. The results indicated that patulin adsorption capability of yeast was influenced by cell surface areas, volume, and cell wall thickness, as well as 1,3-β-glucan content. Among these factors, cell wall thickness and 1,3-β-glucan content serve significant functions. The investigation revealed that patulin adsorption capability was mainly affected by the three-dimensional network structure of the cell wall composed of 1,3-β-glucan. Finally, patulin adsorption in commercial kiwi fruit juice was investigated, and the results indicated that yeast cells could adsorb patulin from commercial kiwi fruit juice efficiently. This study can potentially simulate in vitro cell walls to enhance patulin adsorption capability and successfully apply to fruit juice industry. PMID:26295574

  8. Grazer-induced morphological defense in Scenedesmus obliquus is affected by competition against Microcystis aeruginosa

    PubMed Central

    Zhu, Xuexia; Wang, Jun; Lu, Yichun; Chen, Qinwen; Yang, Zhou

    2015-01-01

    The green alga Scenedesmus is known for its phenotypic plasticity in response to grazing risk. However, the benefits of colony formation induced by infochemicals from zooplankton should come with costs. That is, a tradeoff in benefit-to-cost ratios is likely under complex environmental conditions. In this study, we hypothesized that the coexistence of Scenedesmus and its competitors decreases the formation of anti-grazer colonies in Scenedesmus. Results demonstrated that the presence of a competitor Microcystis aeruginosa inhibited inducible defensive colony formation of Scenedesmus obliquus, and the established defensive colonies negatively affected the competitive ability of S. obliquus. The proportion of induced defensive colonies in cultures was dependent on the relative abundance of competitors. Under low competition intensity, large amount of eight-celled colonies were formed but at the cost of decreased competitive inhibition on M. aeruginosa. By contrast, defensive colony formation of S. obliquus slacked in the presence of high competition intensity to maintain a high displacement rate (competitive ability). In conclusion, S. obliquus exhibited different responses to potential grazing pressure under different intensities of competition, i.e., Scenedesmus morphological response to grazing infochemicals was affected by competition against Microcystis. PMID:26224387

  9. Fractal morphology of Beta vulgaris L. cell suspension culture permeabilized with Triton X-100®

    NASA Astrophysics Data System (ADS)

    Arenas-Ocampo, M.; Alamilla-Beltrán, L.; Vanegas-Espinoza, P. E.; Camacho-Díaz, B. H.; Campos-Mendiola, R.; Gutiérrez-López, G.; Jiménez-Aparicio, A.

    2012-02-01

    In this work, morphology of Beta vulgaris L. cells permeabilized with 0.7mM of Triton X-100® was evaluated using digital image processing and concepts of fractal dimension (perimeter- area relations). Important morphometric changes were found when the contact-time with chemical agent was increased. The size of cells decreased, the cells lost the roundness and their shape was more sinuous; this behaviour was a result of a probable shrinkage caused by the excess of exposure with the permeabilization agent. Morphology of B. vulgaris cells after permeabilization, exhibited a fractal nature since the slope of the ratio of the logarithm of the perimeter vs logarithm of the area was higher than unit. Fractal geometry of the cell morphology was affected as a result of the exposure to Triton X-100®. Those changes can be attributed to the loss of turgor and structure of the cell wall.

  10. Interactions of neurons with topographic nano cues affect branching morphology mimicking neuron-neuron interactions.

    PubMed

    Baranes, Koby; Kollmar, Davida; Chejanovsky, Nathan; Sharoni, Amos; Shefi, Orit

    2012-08-01

    We study the effect of topographic nano-cues on neuronal growth-morphology using invertebrate neurons in culture. We use photolithography to fabricate substrates with repeatable line-pattern ridges of nano-scale heights of 10-150 nm. We plate leech neurons atop the patterned-substrates and compare their growth pattern to neurons plated atop non-patterned substrates. The model system allows us the analysis of single neurite-single ridge interactions. The use of high resolution electron microscopy reveals small filopodia processes that attach to the line-pattern ridges. These fine processes, that cannot be detected in light microscopy, add anchoring sites onto the side of the ridges, thus additional physical support. These interactions of the neuronal process dominantly affect the neuronal growth direction. We analyze the response of the entire neuronal branching tree to the patterned substrates and find significant effect on the growth patterns compared to non-patterned substrates. Moreover, interactions with the nano-cues trigger a growth strategy similarly to interactions with other neuronal cells, as reflected in their morphometric parameters. The number of branches and the number of neurites originating from the soma decrease following the interaction demonstrating a tendency to a more simplified neuronal branching tree. The effect of the nano-cues on the neuronal function deserves further investigation and will strengthen our understanding of the interplay between function and form.

  11. Morphological study of endothelial cells during freezing

    NASA Astrophysics Data System (ADS)

    Zhang, A.; Xu, L. X.; Sandison, G. A.; Cheng, S.

    2006-12-01

    Microvascular injury is recognized as a major tissue damage mechanism of ablative cryosurgery. Endothelial cells lining the vessel wall are thought to be the initial target of freezing. However, details of this injury mechanism are not yet completely understood. In this study, ECMatrix™ 625 was used to mimic the tumour environment and to allow the endothelial cells cultured in vitro to form the tube-like structure of the vasculature. The influence of water dehydration on the integrity of this structure was investigated. It was found that the initial cell shape change was mainly controlled by water dehydration, dependent on the cooling rate, resulting in the shrinkage of cells in the direction normal to the free surface. As the cooling was prolonged and temperature was lowered, further cell shape change could be induced by the chilling effects on intracellular proteins, and focal adhesions to the basement membrane. Quantitative analysis showed that the freezing induced dehydration greatly enhanced the cell surface stresses, especially in the axial direction. This could be one of the major causes of the final breaking of the cell junction and cell detachment.

  12. Physiology of morphologically identified cells of the bullfrog fungiform papilla.

    PubMed

    Takeuchi, H; Tsunenari, T; Kurahashi, T; Kaneko, A

    2001-09-17

    Voltage-gated ionic current and the response to quinine were studied on the four types of morphologically identified taste cells of the bullfrog fungiform papilla by whole-cell patch clamp recording with a Lucifer yellow-filled pipette. Dye-coupled type Ia cells (mucous cells) did not show voltage-activated currents. Type Ib cells (wing cells) characterized by the fin-like processes, type II cells (rod cells) having a thick straight dendrite running to the surface and type III cells with a thin dendrite had voltage-gated sodium (INa) and potassium currents (IK) and generated action potentials. The amplitude of INa was significantly larger in type Ib and II cells than in type III cells. Type Ib and II cells responded to quinine but Type III cells did not.

  13. AQP1 expression alterations affect morphology and water transport in Schwann cells and hypoxia-induced up-regulation of AQP1 occurs in a HIF-1α-dependent manner.

    PubMed

    Zhang, J; Xiong, Y; Lu, L X; Wang, H; Zhang, Y F; Fang, F; Song, Y L; Jiang, H

    2013-11-12

    Aquaporin-1 (AQP1) is the principle water channel in the peripheral nervous system (PNS) and is specifically localized to Schwann cells in the PNS. However, the pathophysiological role of AQP1 in peripheral nerves is poorly understood. Here, we utilized RNA interference by lentiviral transduction to specifically down-regulate AQP1 expression and a lentiviral overexpression protocol to up-regulate AQP1 expression, in primary Schwann cell cultures. AQP1 gene silencing resulted in a cell shrinkage phenotype, while AQP1 gene overexpression caused a cell swelling phenotype, as validated by cell volume determinations. Secondly, we utilized an in vitro hypoxia model in Schwann cells to mimic in vivo facial nerve injury. We demonstrated that AQP1 expression was induced within 8h following hypoxia injury in vitro, and that AQP1 knockdown (KD) caused the cells to resist edema following hypoxia. Finally, we investigated the hypoxic regulation of the AQP1 gene, as well as the involvement of Hypoxia-inducible factor-1α (HIF-1α) in AQP1 modulation and we found that KD of HIF-1α decreased hypoxia-dependent induction of endogenous AQP1 expression at both the mRNA and protein levels. Taken together, these results indicate that (1) AQP1 is an important factor responsible for the fast water transport of cultured Schwann cells and is involved in cell plasticity; (2) AQP1 alterations may be a primary factor in hypoxia-induced peripheral nerve edema; (3) HIF-1α participates in the hypoxic induction of the AQP1 gene; (4) AQP1 inhibition might provide a new therapeutic alternative for the treatment of some forms of peripheral nerve edema.

  14. Effects of hypergravity on adipose-derived stem cell morphology, mechanical property and proliferation

    SciTech Connect

    Tavakolinejad, Alireza; Rabbani, Mohsen; Janmaleki, Mohsen

    2015-08-21

    Alteration in specific inertial conditions can lead to changes in morphology, proliferation, mechanical properties and cytoskeleton of cells. In this report, the effects of hypergravity on morphology of Adipose-Derived Stem Cells (ADSCs) are indicated. ADSCs were repeatedly exposed to discontinuous hypergravity conditions of 10 g, 20 g, 40 g and 60 g by utilizing centrifuge (three times of 20 min exposure, with an interval of 40 min at 1 g). Cell morphology in terms of length, width and cell elongation index and cytoskeleton of actin filaments and microtubules were analyzed by image processing. Consistent changes observed in cell elongation index as morphological change. Moreover, cell proliferation was assessed and mechanical properties of cells in case of elastic modulus of cells were evaluated by Atomic Force Microscopy. Increase in proliferation and decrease in elastic modulus of cells are further results of this study. Staining ADSC was done to show changes in cytoskeleton of the cells associated to hypergravity condition specifically in microfilament and microtubule components. After exposing to hypergravity, significant changes were observed in microfilaments and microtubule density as components of cytoskeleton. It was concluded that there could be a relationship between changes in morphology and MFs as the main component of the cells. - Highlights: • Hypergravity (10 g, 20 g, 40 g and 60 g) affects on adipose derived stem cells (ADSCs). • ADSCs after exposure to the hypergravity are more slender. • The height of ADSCs increases in all test groups comparing their control group. • Hypergravity decreases ADSCs modulus of elasticity and cell actin fiber content. • Hypergravity enhances proliferation rate of ADSCs.

  15. Effect of surface potential on epithelial cell adhesion, proliferation and morphology.

    PubMed

    Chang, Hsun-Yun; Kao, Wei-Lun; You, Yun-Wen; Chu, Yi-Hsuan; Chu, Kuo-Jui; Chen, Peng-Jen; Wu, Chen-Yi; Lee, Yu-Hsuan; Shyue, Jing-Jong

    2016-05-01

    Cell adhesion is the basis of individual cell survival, division and motility. Hence, understanding the effects that the surface properties have on cell adhesion, proliferation and morphology are crucial. In particular, surface charge/potential has been identified as an important factor that affects cell behavior. However, how cells respond to incremental changes in surface potential remains unclear. By using binary self-assembled monolayer (SAM) modified Au surfaces that are similar in mechanical/chemical properties and provide a series of surface potentials, the effect of surface potential on the behavior of cells can be studied. In this work, the effect of surface potential on epithelial cells, including human embryonic kidney (HEK293T) and human hepatocellular carcinoma (HepG2), were examined. The results showed that the adhesion density of epithelial cells increased with increasing surface potential, which is similar to but varied more significantly compared with fibroblasts. The proliferation rate is found to be independent of surface potential in both cell types. Furthermore, epithelial cells show no morphological change with respect to surface potential, whereas the morphology of the fibroblasts clearly changed with the surface potential. These differences between the cell types were rationalized by considering the difference in extracellular matrix composition. Laminin-dominant epithelial cells showed higher adhesion density and less morphological change than did fibronectin-dominant fibroblasts because the more significant adsorption of positively charged laminin on the surface enhanced the adhesion of epithelial cells. In contrast, due to the dominance of negatively charged fibronectin that adsorbed weakly on the surface, fibroblasts had to change their morphology to fit the inhomogeneous fibronectin-adsorbed area.

  16. Rheumatoid synovial cells from intact joints. Morphology, growth, and polykaryocytosis.

    PubMed

    Clarris, B J; Fraser, J R; Moran, C J; Muirden, K D

    1977-08-01

    Synovial cell lines were isolated by instillation of trypsin or chymotrypsin into intact knee joints of patients with persistent rheumatoid effusions resistant to conventional therapy. Morphology and growth in the primary phase were compared with rheumatoid cells isolated from excised synovium and nonrheumatoid synovial cells obtained from intact joints of cadavers or amputated limbs. Cell populations from all sources included varying proportions of macrophage-like and fibroblast-like cells, with only 1-3% multinucleated cells. In medium supplemented with calf serum alone, rheumatoid cells from intact joints showed negligible changes in morphology. However, in the presence of nonrheumatoid, autologous rheumatoid or homologous rheumatoid serum a rapid increase occurred in size of the macrophage-like cells and numbers of polykaryocytes, including some giant syncytial cells. These effects were directly proportional to serum concentration and were identical in fresh or heat-inactivated serum. In most of these rheumatoid cell lines no multiplication occurred, regardless of serum type or concentration. In rheumatoid synovial cells from excised synovium, human serum induced both polykaryocytosis and rapid growth of fibroblasts. Nonrheumatoid synovial cells grew rapidly but few polykaryocytes developed, mostly with less than 6 nuclei. Evidence of viral infection in rheumatoid synovial cells was sought by electron microscopy after stimulation of polykaryocytosis by human serum. In one of the cultures many cells were found with intranuclear particles possessing characteristics of the adenovirus group. PMID:901027

  17. Counting white blood cells using morphological granulometries

    NASA Astrophysics Data System (ADS)

    Theera-Umpon, Nipon; Gader, Paul D.

    2000-04-01

    We describe a modification of the mixture proportion estimation algorithm based on the granulometric mixing theorem. The modified algorithm is applied to the problem of counting different types of white blood cells in bone marrow images. In principle, the algorithm can be used to count the proportion of cells in each class without explicitly segmenting and classifying them. The direct application of the original algorithm does not converge well for more than two classes. The modified algorithm uses prior statistics to initially segment the mixed pattern spectrum and then applies the one-primitive estimation algorithm to each initial component. Applying the algorithm to one class at a time results in better convergence. The counts produced by the modified algorithm on six classes of cells--myeloblast, promyelocyte, myelocyte, metamyelocyte, band, and PolyMorphoNuclear--are very close to the human expert's numbers; the deviation of the algorithm counts is similar to the deviation of counts produced by human experts. The important technical contributions are that the modified algorithm uses prior statistics for each shape class in place or prior knowledge of the total number of objects in an image, and it allows for more than one primitive from each class.

  18. [Changes of blood cell's morphological characteristics and their clinical significance].

    PubMed

    Mestiashvili, I G

    2013-01-01

    The purpose of this article is the interpretation of the clinical value of blood cells' (erythrocytes, leukocytes, thrombocytes) morphological characteristics changes taking into the consideration results of our own long-term (1982-2012) investigations as well as literature data. Patients with hematological and nonhematological diseases as well as large groups of Georgia population (about 30.000 persons, in age 2-81) were observed. By Gimsa-Romanovsky method stained blood smears were studied in light microscope. Changes of blood cells' morphological characteristics, observed by us and known in the literature, as well as ones revealed at first in this work, are presented according to the three branches of hemopoiesis. Possible changes of blood cells' morphological characteristics are described in details. Concrete changes in concrete diseases are presented in eight tables. Discussing the results the fact of the revealing of blood cells' morphological changes in some diseases, different by etiology, pathogenesis, clinical symptomatology and treatment, is accentuated. Some examples of the differentiation of different diseases by means of that nuances, which are revealed in microscope and which are not noticeable for the modern techniques, are presented. The significance of some specific changes of blood cells' morphological characteristics for the doubtless diagnostic of some concrete anomalies is shown. The list of changes which are revealed in this work but are not founded in the literature is presented. The conclusion is drawn that the nuances of blood cells` morphological changes revealed in light microscope allow to narrow the spectrum of additional investigations for the establishment of diagnosis.

  19. [MORPHOLOGY OF NCTC CELLS ONE DAY AFTER RESEEDING].

    PubMed

    Petrov, Yu P; Tsupkina, N V

    2016-01-01

    Development of regenerative medicine based on the use of stem cells is substantially dependent on the prediction of the changes that the cells undergo after culturing them in vitro. Therefore, the accumulation of knowledge in the field, which can be denoted as biology of cells in culture, is of special importance. Features of functioning cells in vitro is better to study in the permanent cells lines as their morphological and functional characteristics in numerous passages can be regarded as the result of adaptation of cells to grow outside the body. The aim of the present study was to test whether there is a relationship between the density of the cell culture prior to the formation of a monolayer of cells and morphometric parameters of the cells. The NCTC fibroblast-like cells (clone 929 were examined one day after reseeding. By this time, the culture density was such that there was virtually no direct contact between the cells. The cell area, spreading and polarization coefficients were used to characterize the cells. It has been shown that, in the same culture flask, the cells in the areas with a higher density of cells are smaller than in areas of lower density. At the same time, polarization of cells increases by increasing the cell density. Such cell reaction may be the result of the remote transfer of information between the cells. Analysis of the data obtained allows us to assume that the change in shape of the cells is related to early steps of monolayer formation. PMID:27220250

  20. Supramolecular Approaches to Nanoscale Morphological Control in Organic Solar Cells.

    PubMed

    Haruk, Alexander M; Mativetsky, Jeffrey M

    2015-06-11

    Having recently surpassed 10% efficiency, solar cells based on organic molecules are poised to become a viable low-cost clean energy source with the added advantages of mechanical flexibility and light weight. The best-performing organic solar cells rely on a nanostructured active layer morphology consisting of a complex organization of electron donating and electron accepting molecules. Although much progress has been made in designing new donor and acceptor molecules, rational control over active layer morphology remains a central challenge. Long-term device stability is another important consideration that needs to be addressed. This review highlights supramolecular strategies for generating highly stable nanostructured organic photovoltaic active materials by design.

  1. Defect-Mediated Morphologies in Growing Cell Colonies.

    PubMed

    Doostmohammadi, Amin; Thampi, Sumesh P; Yeomans, Julia M

    2016-07-22

    Morphological trends in growing colonies of living cells are at the core of physiological and evolutionary processes. Using active gel equations, which include cell division, we show that shape changes during the growth can be regulated by the dynamics of topological defects in the orientation of cells. The friction between the dividing cells and underlying substrate drives anisotropic colony shapes toward more isotropic morphologies, by mediating the number density and velocity of topological defects. We show that the defects interact with the interface at a specific interaction range, set by the vorticity length scale of flows within the colony, and that the cells predominantly reorient parallel to the interface due to division-induced active stresses. PMID:27494503

  2. Defect-Mediated Morphologies in Growing Cell Colonies

    NASA Astrophysics Data System (ADS)

    Doostmohammadi, Amin; Thampi, Sumesh P.; Yeomans, Julia M.

    2016-07-01

    Morphological trends in growing colonies of living cells are at the core of physiological and evolutionary processes. Using active gel equations, which include cell division, we show that shape changes during the growth can be regulated by the dynamics of topological defects in the orientation of cells. The friction between the dividing cells and underlying substrate drives anisotropic colony shapes toward more isotropic morphologies, by mediating the number density and velocity of topological defects. We show that the defects interact with the interface at a specific interaction range, set by the vorticity length scale of flows within the colony, and that the cells predominantly reorient parallel to the interface due to division-induced active stresses.

  3. Whole organ, venation and epidermal cell morphological variations are correlated in the leaves of Arabidopsis mutants.

    PubMed

    Pérez-Pérez, José Manuel; Rubio-Díaz, Silvia; Dhondt, Stijn; Hernández-Romero, Diana; Sánchez-Soriano, Joaquín; Beemster, Gerrit T S; Ponce, María Rosa; Micol, José Luis

    2011-12-01

    Despite the large number of genes known to affect leaf shape or size, we still have a relatively poor understanding of how leaf morphology is established. For example, little is known about how cell division and cell expansion are controlled and coordinated within a growing leaf to eventually develop into a laminar organ of a definite size. To obtain a global perspective of the cellular basis of variations in leaf morphology at the organ, tissue and cell levels, we studied a collection of 111 non-allelic mutants with abnormally shaped and/or sized leaves, which broadly represent the mutational variations in Arabidopsis thaliana leaf morphology not associated with lethality. We used image-processing techniques on these mutants to quantify morphological parameters running the gamut from the palisade mesophyll and epidermal cells to the venation, whole leaf and rosette levels. We found positive correlations between epidermal cell size and leaf area, which is consistent with long-standing Avery's hypothesis that the epidermis drives leaf growth. In addition, venation parameters were positively correlated with leaf area, suggesting that leaf growth and vein patterning share some genetic controls. Positional cloning of the genes affected by the studied mutations will eventually establish functional links between genotypes, molecular functions, cellular parameters and leaf phenotypes.

  4. Morphology control of zinc regeneration for zinc-air fuel cell and battery

    NASA Astrophysics Data System (ADS)

    Wang, Keliang; Pei, Pucheng; Ma, Ze; Xu, Huachi; Li, Pengcheng; Wang, Xizhong

    2014-12-01

    Morphology control is crucial both for zinc-air batteries and for zinc-air fuel cells during zinc regeneration. Zinc dendrite should be avoided in zinc-air batteries and zinc pellets are yearned to be formed for zinc-air fuel cells. This paper is mainly to analyze the mechanism of shape change and to control the zinc morphology during charge. A numerical three-dimensional model for zinc regeneration is established with COMSOL software on the basis of ionic transport theory and electrode reaction electrochemistry, and some experiments of zinc regeneration are carried out. The deposition process is qualitatively analyzed by the kinetics Monte Carlo method to study the morphological change from the electrocrystallization point of view. Morphological evolution of deposited zinc under different conditions of direct currents and pulse currents is also investigated by simulation. The simulation shows that parametric variables of the flowing electrolyte, the surface roughness and the structure of the electrode, the charging current and mode affect morphological evolution. The uniform morphology of deposited zinc is attained at low current, pulsating current or hydrodynamic electrolyte, and granular morphology is obtained by means of an electrode of discrete columnar structure in combination with high current and flowing electrolyte.

  5. Morphological effect of oscillating magnetic nanoparticles in killing tumor cells

    PubMed Central

    2014-01-01

    Forced oscillation of spherical and rod-shaped iron oxide magnetic nanoparticles (MNPs) via low-power and low-frequency alternating magnetic field (AMF) was firstly used to kill cancer cells in vitro. After being loaded by human cervical cancer cells line (HeLa) and then exposed to a 35-kHz AMF, MNPs mechanically damaged cell membranes and cytoplasm, decreasing the cell viability. It was found that the concentration and morphology of the MNPs significantly influenced the cell-killing efficiency of oscillating MNPs. In this preliminary study, when HeLa cells were pre-incubated with 100 μg/mL rod-shaped MNPs (rMNP, length of 200 ± 50 nm and diameter of 50 to 120 nm) for 20 h, MTT assay proved that the cell viability decreased by 30.9% after being exposed to AMF for 2 h, while the cell viability decreased by 11.7% if spherical MNPs (sMNP, diameter of 200 ± 50 nm) were used for investigation. Furthermore, the morphological effect of MNPs on cell viability was confirmed by trypan blue assay: 39.5% rMNP-loaded cells and 15.1% sMNP-loaded cells were stained after being exposed to AMF for 2 h. It was also interesting to find that killing tumor cells at either higher (500 μg/mL) or lower (20 μg/mL) concentration of MNPs was less efficient than that achieved at 100 μg/mL concentration. In conclusion, the relatively asymmetric morphological rod-shaped MNPs can kill cancer cells more effectively than spherical MNPs when being exposed to AMF by virtue of their mechanical oscillations. PMID:24872797

  6. Factors Affecting the Morphology of Pb-Based Glass Frit Coated with Ag Material Prepared by Electroless Silver Plating

    NASA Astrophysics Data System (ADS)

    Huang, Bei; Gan, Weiping; Zhou, Jian; Li, Yingfen; Lin, Tao; Liu, Xiaogang

    2014-05-01

    Pb-based glass frit coated with nanosilver material for Si solar cell applications has been directly prepared by electroless silver plating. Activation of the glass frit was accomplished by using glycol, with the aim of reducing the silver ions to elemental silver on the surface of the glass frit. Electroless silver plating onto the glass frit was successfully realized using two kinds of electroless plating bath. However, the morphology of the composite powder greatly affected the modality, sheet resistance, series resistance, and photoelectric conversion efficiency of the conducting silver films. We found that the activation temperature affected the number and distribution of silver nanoparticles. Meanwhile, the average grain size of the silver particles and the silver content in the Pb-based glass frit coated with Ag material could be controlled by adjusting the pH value and loading capacity, respectively, during plating.

  7. Morphologic changes in basal cells during repair of tracheal epithelium.

    PubMed Central

    Wang, C. Z.; Evans, M. J.; Cox, R. A.; Burke, A. S.; Zhu, Q.; Herndon, D. N.; Barrow, R. E.

    1992-01-01

    Basal cells are differentiated with respect to junctional adhesion mechanisms and play a role in attachment of columnar epithelium to the basal lamina. Although much is known about nonciliated and ciliated cell differentiation during the repair process after injury, little is known about the basal cell. We studied the morphology of basal cells and quantitated junctional adhesion structures during repair of tracheal epithelium exposed to toxic cotton smoke. Ten adult ewes were given a smoke injury to a portion of the upper cervical trachea and were killed at 4, 6, 8, 10, and 18 days after injury for morphometric studies. At 4 days, there was a stratified reparative epithelium over the basal lamina, which was two to four cells in depth. The basal cells were identified by their hemidesmosome (HD) attachment to the basal lamina. Basal cells were about 69% larger than controls and flattened rather than columnar. The amount of HD attachment was 192% greater than controls. In contrast, volume density of cytokeratin filaments had decreased about 47%. Basal cells had returned to normal numbers and size and a columnar shape by day 18. The amount of desmosome (D) and HD attachment and volume density of cytokeratins had also reached control levels by day 18. These data indicate that morphology of basal cells changes during the initial stages of reparative regeneration but returns to normal by 18 days. Morphologic changes appear to reflect changes in size of the cell associated with cell division rather than differentiation of recently divided basal cells. Images Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 PMID:1381564

  8. Prophylactic pinning for slipped capital femoral epiphysis: does it affect proximal femoral morphology?

    PubMed

    Cousins, Gerard R; Campbell, Donald M; Wilson, Neil I L; Maclean, Jamie G B

    2016-05-01

    This study was designed to determine whether prophylactic pinning of the unaffected hip in unilateral slipped capital femoral epiphysis affects the proximal femoral morphology. Twenty-four hips prophylactically pinned were compared with 26 cases observed. The articulotrochanteric distance (ATD) and the trochanteric-trochanteric distance (TTD) were measured. Postoperative radiographs were compared with final follow-up radiographs. The final TTD : ATD ratio was higher (P=0.048) in the pinned group, suggesting relative coxa vara/breva. There was a smaller difference between the two hips in the prophylactically pinned group (0.7) as opposed to those observed (1.47). Prophylactic pinning does not cause growth to stop immediately but alters the proximal femoral morphology.

  9. Cell morphology and flagellation of nitrogen-fixing spirilla.

    PubMed

    Hegazi, N A; Vlassak, K

    1979-01-01

    Twenty isolates of N2-fixing spirilla were isolated from the rhizosphere of maize and sugar cane grown in Egyptian and Belgian soils. Electron microscopy distinguished two morphological groups. The first includes short and thick curved rods with an unipolar flagellum while cells of the second group are much longer with the typical appearance of spiral cells and most probably possess a bipolar tuft of flagella. PMID:527912

  10. Tendon cell outgrowth rates and morphology associated with kevlar-49.

    PubMed

    Zimmerman, M; Gordon, K E

    1988-12-01

    A rat tendon cell model was used to evaluate the in vitro biocompatibility of kevlar-49. The cell response to kevlar was compared to carbon AS-4 and nylon sutures. Three trials were run and cell growth rates were statistically similar for all the materials tested. A separate experiment was conducted in which the same fiber materials were placed in the same Petri dish. Again, the rates were similar for each material. Finally, the cells were observed with a scanning electron microscope, and the three classic cell morphologies associated with this tendon cell model were observed. Also, cellular attachment to the fiber and cellular encapsulation of the fiber were identical for the three materials tested. Kevlar-49 proved to be comparable to carbon AS4 and nylon sutures in terms of cellular response and cell outgrowth rates.

  11. Mammosomatotroph cell adenoma of the human pituitary: a morphologic entity.

    PubMed

    Horvath, E; Kovacs, K; Killinger, D W; Smyth, H S; Weiss, M H; Ezrin, C

    1983-01-01

    Nine cases of a hitherto undescribed morphologic entity, termed mammosomatotroph cell adenoma of the human pituitary, are reported. These tumors, occurring mostly in men, are invariably associated with acromegaly (or gigantism) and high-normal or slightly elevated blood prolactin levels, and it cannot be distinguished clinically from well-differentiated growth hormone cell or mixed growth hormone cell-prolactin cell adenomas. They show a slow growth rate and usually exhibit a diffuse pattern and intense cytoplasmic acidophilia by histology. The immunoperoxidase technique detects both growth hormone and prolactin within the same cells. Electron microscopy reveals monomorphous tumors with a fine structure markedly similar to that of well-differentiated, densely granulated growth hormone cell adenomas. An added feature and diagnostic marker of mammosomatotroph cell adenoma is the presence of extracellular deposits of secretory material. One tumor shows a marked abnormality of hormone packaging and storage, resulting in the cytoplasmic accumulation of pleomorphic bodies containing semicrystalline secretory material. PMID:6402839

  12. Folic acid supplementation for 4 weeks affects liver morphology in aged rats.

    PubMed

    Roncalés, María; Achón, María; Manzarbeitia, Félix; Maestro de las Casas, Carmen; Ramírez, Carmen; Varela-Moreiras, Gregorio; Pérez-Miguelsanz, Julia

    2004-05-01

    Several countries have approved universal folic acid (FA) fortification to prevent neural tube defects and/or high homocysteine levels; this has led to a chronic intake of FA. Traditionally, the vitamin is considered to be safe and nontoxic, except for the potential masking of vitamin B-12 deficiency. Recent reports from our laboratories showed several effects of high-dose folate supplementation in rats. In this work, we compared the effect of FA on the liver of weanling (3 wk) and aged (18 mo) male rats fed either a diet supplemented with 40 mg FA/kg diet or a control diet (1 mg FA/kg diet) for 4 wk. FA supplementation did not alter serum aspartate aminotransferase, alanine aminotransferase, urea, glucose oxidase, total bilirubin, or uric acid. Routine histological staining as well as immunohistochemistry with proliferating cell nuclear antibody for dividing cells, and cytokeratin-8 against bile ductal cells, showed that aged, supplemented rats had the same number of hepatocytes as both control and supplemented weanling rats, and tended to have more (17%, P = 0.07) hepatocytes than aged, control rats. Moreover, the bile duct cells of aged, control rats proliferated and transformed into cholestatic rosettes at a higher frequency than in aged, supplemented rats. The morphology of the liver in weanling rats was similar in both diet groups, and comparable to the supplemented, aged rats, thus indicating that a high intake of FA improves normal liver morphology in livers of aged rats.

  13. Physical parameters affecting living cells in space

    NASA Astrophysics Data System (ADS)

    Langbein, Dieter

    The question is posed: Why does a living cell react to the absence of gravity? What sensors may it have? Does it note pressure, sedimentation, convection, or other parameters? If somewhere in a liquid volume sodium ions are replaced by potassium ions, the density of the liquid changes locally: the heavier regions sink, the lighter regions rise. This may contribute to species transport, to the metabolism. Under microgravity this mechanism is strongly reduced. On the other hand, other reasons for convection like thermal and solutal interface convection are left. Do they affect species transport? Another important effect of gravity is the hydrostatic pressure. On the macroscopic side, the pressure between our head and feet changes by 0.35 atmospheres. On the microscopic level the hydrostatic pressure on the upper half of a cell membrane is lower than on the lower half. This, by affecting the ion transport through the membrane, may change the surrounding electric potential. It has been suggested to be one of the reasons for graviperception. Following the discussion of these and other effects possibly important in life sciences in space, an order of magnitude analysis of the residual accelerations tolerable during experiments in materials sciences is outlined. In the field of life sciences only rough estimates are available at present.

  14. Dynamic and reversible surface topography influences cell morphology.

    PubMed

    Kiang, Jennifer D; Wen, Jessica H; del Álamo, Juan C; Engler, Adam J

    2013-08-01

    Microscale and nanoscale surface topography changes can influence cell functions, including morphology. Although in vitro responses to static topography are novel, cells in vivo constantly remodel topography. To better understand how cells respond to changes in topography over time, we developed a soft polyacrylamide hydrogel with magnetic nickel microwires randomly oriented in the surface of the material. Varying the magnetic field around the microwires reversibly induced their alignment with the direction of the field, causing the smooth hydrogel surface to develop small wrinkles; changes in surface roughness, ΔRRMS , ranged from 0.05 to 0.70 μm and could be oscillated without hydrogel creep. Vascular smooth muscle cell morphology was assessed when exposed to acute and dynamic topography changes. Area and shape changes occurred when an acute topographical change was imposed for substrates exceeding roughness of 0.2 μm, but longer-term oscillating topography did not produce significant changes in morphology irrespective of wire stiffness. These data imply that cells may be able to use topography changes to transmit signals as they respond immediately to changes in roughness.

  15. Bacillus subtilis α-Phosphoglucomutase Is Required for Normal Cell Morphology and Biofilm Formation†

    PubMed Central

    Lazarevic, Vladimir; Soldo, Blazenka; Médico, Noël; Pooley, Harold; Bron, Sierd; Karamata, Dimitri

    2005-01-01

    Mutations designated gtaC and gtaE that affect α-phosphoglucomutase activity required for interconversion of glucose 6-phosphate and α-glucose 1-phosphate were mapped to the Bacillus subtilis pgcA (yhxB) gene. Backcrossing of the two mutations into the 168 reference strain was accompanied by impaired α-phosphoglucomutase activity in the soluble cell extract fraction, altered colony and cell morphology, and resistance to phages φ29 and ρ11. Altered cell morphology, reversible by additional magnesium ions, may be correlated with a deficiency in the membrane glycolipid. The deficiency in biofilm formation in gtaC and gtaE mutants may be attributed to an inability to synthesize UDP-glucose, an important intermediate in a number of cell envelope biosynthetic processes. PMID:15640167

  16. Three-Dimensional Numerical Model of Cell Morphology during Migration in Multi-Signaling Substrates

    PubMed Central

    Mousavi, Seyed Jamaleddin; Hamdy Doweidar, Mohamed

    2015-01-01

    Cell Migration associated with cell shape changes are of central importance in many biological processes ranging from morphogenesis to metastatic cancer cells. Cell movement is a result of cyclic changes of cell morphology due to effective forces on cell body, leading to periodic fluctuations of the cell length and cell membrane area. It is well-known that the cell can be guided by different effective stimuli such as mechanotaxis, thermotaxis, chemotaxis and/or electrotaxis. Regulation of intracellular mechanics and cell’s physical interaction with its substrate rely on control of cell shape during cell migration. In this notion, it is essential to understand how each natural or external stimulus may affect the cell behavior. Therefore, a three-dimensional (3D) computational model is here developed to analyze a free mode of cell shape changes during migration in a multi-signaling micro-environment. This model is based on previous models that are presented by the same authors to study cell migration with a constant spherical cell shape in a multi-signaling substrates and mechanotaxis effect on cell morphology. Using the finite element discrete methodology, the cell is represented by a group of finite elements. The cell motion is modeled by equilibrium of effective forces on cell body such as traction, protrusion, electrostatic and drag forces, where the cell traction force is a function of the cell internal deformations. To study cell behavior in the presence of different stimuli, the model has been employed in different numerical cases. Our findings, which are qualitatively consistent with well-known related experimental observations, indicate that adding a new stimulus to the cell substrate pushes the cell to migrate more directionally in more elongated form towards the more effective stimuli. For instance, the presence of thermotaxis, chemotaxis and electrotaxis can further move the cell centroid towards the corresponding stimulus, respectively, diminishing the

  17. Citral exerts its antifungal activity against Penicillium digitatum by affecting the mitochondrial morphology and function.

    PubMed

    Zheng, Shiju; Jing, Guoxing; Wang, Xiao; Ouyang, Qiuli; Jia, Lei; Tao, Nengguo

    2015-07-01

    This work investigated the effect of citral on the mitochondrial morphology and function of Penicillium digitatum. Citral at concentrations of 2.0 or 4.0 μL/mL strongly damaged mitochondria of test pathogen by causing the loss of matrix and increase of irregular mitochondria. The deformation extent of the mitochondria of P. digitatum enhanced with increasing concentrations of citral, as evidenced by a decrease in intracellular ATP content and an increase in extracellular ATP content of P. digitatum cells. Oxygen consumption showed that citral resulted in an inhibition in the tricarboxylic acid cycle (TCA) pathway of P. digitatum cells, induced a decrease in activities of citrate synthetase, isocitrate dehydrogenase, α-ketoglutarate dehydrogenase, succinodehydrogenase and the content of citric acid, while enhancing the activity of malic dehydrogenase in P. digitatum cells. Our present results indicated that citral could damage the mitochondrial membrane permeability and disrupt the TCA pathway of P. digitatum.

  18. Endothelial Cell Morphology and Migration are Altered by Changes in Gravitational Fields

    NASA Technical Reports Server (NTRS)

    Melhado, Caroline; Sanford, Gary; Harris-Hooker, Sandra

    1997-01-01

    Endothelial cell migration is important to vascular wall regeneration following injury or stress. However, the mechanism(s) governing this response is not well understood. The microgravity environment of space may complicate the response of these cells to injury. To date, there are no reports in this area. We examined how bovine aortic (BAEC) and pulmonary (BPEC) endothelial cells respond to denudation injury under hypergravity (HGrav) and simulated microgravity (MGrav), using image analysis. In 10% FBS, the migration of confluent BAEC and BPEC into the denuded area was not affected by HGrav or MGrav. However, in low FBS (0.5%), signficantly retarded migration under MGrav, and increased migration under HGrav was found. MGrav also decreased the migration of postconfluent BPEC while HGrav showed no difference. Both MGrav and HGrav strongly decreased the migration of postconfluent BAEC. Also, both cell lines showed significant morphological changes by scanning electron microscopy. These studies indicate that endothelial cell function is affected by changes in gravity.

  19. Urban habitat complexity affects species richness but not environmental filtering of morphologically-diverse ants.

    PubMed

    Ossola, Alessandro; Nash, Michael A; Christie, Fiona J; Hahs, Amy K; Livesley, Stephen J

    2015-01-01

    Habitat complexity is a major determinant of structure and diversity of ant assemblages. Following the size-grain hypothesis, smaller ant species are likely to be advantaged in more complex habitats compared to larger species. Habitat complexity can act as an environmental filter based on species size and morphological traits, therefore affecting the overall structure and diversity of ant assemblages. In natural and semi-natural ecosystems, habitat complexity is principally regulated by ecological successions or disturbance such as fire and grazing. Urban ecosystems provide an opportunity to test relationships between habitat, ant assemblage structure and ant traits using novel combinations of habitat complexity generated and sustained by human management. We sampled ant assemblages in low-complexity and high-complexity parks, and high-complexity woodland remnants, hypothesizing that (i) ant abundance and species richness would be higher in high-complexity urban habitats, (ii) ant assemblages would differ between low- and high-complexity habitats and (iii) ants living in high-complexity habitats would be smaller than those living in low-complexity habitats. Contrary to our hypothesis, ant species richness was higher in low-complexity habitats compared to high-complexity habitats. Overall, ant assemblages were significantly different among the habitat complexity types investigated, although ant size and morphology remained the same. Habitat complexity appears to affect the structure of ant assemblages in urban ecosystems as previously observed in natural and semi-natural ecosystems. However, the habitat complexity filter does not seem to be linked to ant morphological traits related to body size. PMID:26528416

  20. Urban habitat complexity affects species richness but not environmental filtering of morphologically-diverse ants

    PubMed Central

    Nash, Michael A.; Christie, Fiona J.; Hahs, Amy K.; Livesley, Stephen J.

    2015-01-01

    Habitat complexity is a major determinant of structure and diversity of ant assemblages. Following the size-grain hypothesis, smaller ant species are likely to be advantaged in more complex habitats compared to larger species. Habitat complexity can act as an environmental filter based on species size and morphological traits, therefore affecting the overall structure and diversity of ant assemblages. In natural and semi-natural ecosystems, habitat complexity is principally regulated by ecological successions or disturbance such as fire and grazing. Urban ecosystems provide an opportunity to test relationships between habitat, ant assemblage structure and ant traits using novel combinations of habitat complexity generated and sustained by human management. We sampled ant assemblages in low-complexity and high-complexity parks, and high-complexity woodland remnants, hypothesizing that (i) ant abundance and species richness would be higher in high-complexity urban habitats, (ii) ant assemblages would differ between low- and high-complexity habitats and (iii) ants living in high-complexity habitats would be smaller than those living in low-complexity habitats. Contrary to our hypothesis, ant species richness was higher in low-complexity habitats compared to high-complexity habitats. Overall, ant assemblages were significantly different among the habitat complexity types investigated, although ant size and morphology remained the same. Habitat complexity appears to affect the structure of ant assemblages in urban ecosystems as previously observed in natural and semi-natural ecosystems. However, the habitat complexity filter does not seem to be linked to ant morphological traits related to body size. PMID:26528416

  1. Relation of CD30 expression to survival and morphology in large cell B cell lymphomas.

    PubMed Central

    Noorduyn, L A; de Bruin, P C; van Heerde, P; van de Sandt, M M; Ossenkoppele, G J; Meijer, C J

    1994-01-01

    AIMS--To investigate whether CD30 expression is correlated with anaplastic morphology, and whether this correlated with a better survival in large cell B cell lymphomas, as has been described for T cell lymphomas. METHODS--CD30 expression was investigated using frozen sections in a series of 146 large cell B cell lymphomas. Clinical data and follow up information were collected from 25 lymphomas with strong CD30 expression, 30 lymphomas with partial CD30 expression, and a control group of 25 lymphomas which did not express CD30. RESULTS--Morphological distinction between anaplastic and non-anaplastic tumours was difficult. Of the cases with an anaplastic morphology, 50% were CD30 positive, as were 24% of the polymorphic centroblastic B cell lymphomas. Only 65% of the morphologically non-anaplastic tumours were completely CD30 negative. There was no difference in survival among patients with lymphomas expressing CD30 and those that did not. Patients with morphologically anaplastic B cell lymphomas did not differ in their survivals from those with other high grade B cell lymphomas. Clinical stage at presentation was the only variable that was significantly associated with survival. CONCLUSIONS--CD30 expression occurs frequently in large cell B cell lymphomas and is poorly related to anaplastic morphology. Morphological distinction between anaplastic and non-anaplastic tumours is difficult. In contrast to T cell lymphomas, CD30 positive B cell lymphomas do not show a relatively favourable clinical course. The results presented here raise serious doubts as to whether large cell B cell lymphoma, based on the expression of CD30 or anaplastic morphology, can really be termed a separate entity. Images PMID:8132806

  2. Early postnatal respiratory viral infection alters hippocampal neurogenesis, cell fate, and neuron morphology in the neonatal piglet.

    PubMed

    Conrad, Matthew S; Harasim, Samantha; Rhodes, Justin S; Van Alstine, William G; Johnson, Rodney W

    2015-02-01

    Respiratory viral infections are common during the neonatal period in humans, but little is known about how early-life infection impacts brain development. The current study used a neonatal piglet model as piglets have a gyrencephalic brain with growth and development similar to human infants. Piglets were inoculated with porcine reproductive and respiratory syndrome virus (PRRSV) to evaluate how chronic neuroinflammation affects hippocampal neurogenesis and neuron morphology. Piglets in the neurogenesis study received one bromodeoxyuridine injection on postnatal day (PD) 7 and then were inoculated with PRRSV. Piglets were sacrificed at PD 28 and the number of BrdU+ cells and cell fate were quantified in the dentate gyrus. PRRSV piglets showed a 24% reduction in the number of newly divided cells forming neurons. Approximately 15% of newly divided cells formed microglia, but this was not affected by sex or PRRSV. Additionally, there was a sexual dimorphism of new cell survival in the dentate gyrus where males had more cells than females, and PRRSV infection caused a decreased survival in males only. Golgi impregnation was used to characterize dentate granule cell morphology. Sholl analysis revealed that PRRSV caused a change in inner granule cell morphology where the first branch point was extended further from the cell body. Males had more complex dendritic arbors than females in the outer granule cell layer, but this was not affected by PRRSV. There were no changes to dendritic spine density or morphology distribution. These findings suggest that early-life viral infection can impact brain development.

  3. Human Mesenchymal Stem Cell Morphology and Migration on Microtextured Titanium

    PubMed Central

    Banik, Brittany L.; Riley, Thomas R.; Platt, Christina J.; Brown, Justin L.

    2016-01-01

    The implant used in spinal fusion procedures is an essential component to achieving successful arthrodesis. At the cellular level, the implant impacts healing and fusion through a series of steps: first, mesenchymal stem cells (MSCs) need to adhere and proliferate to cover the implant; second, the MSCs must differentiate into osteoblasts; third, the osteoid matrix produced by the osteoblasts needs to generate new bone tissue, thoroughly integrating the implant with the vertebrate above and below. Previous research has demonstrated that microtextured titanium is advantageous over smooth titanium and PEEK implants for both promoting osteogenic differentiation and integrating with host bone tissue; however, no investigation to date has examined the early morphology and migration of MSCs on these surfaces. This study details cell spreading and morphology changes over 24 h, rate and directionality of migration 6–18 h post-seeding, differentiation markers at 10 days, and the long-term morphology of MSCs at 7 days, on microtextured, acid-etched titanium (endoskeleton), smooth titanium, and smooth PEEK surfaces. The results demonstrate that in all metrics, the two titanium surfaces outperformed the PEEK surface. Furthermore, the rough acid-etched titanium surface presented the most favorable overall results, demonstrating the random migration needed to efficiently cover a surface in addition to morphologies consistent with osteoblasts and preosteoblasts. PMID:27243001

  4. Light deprivation delays morphological differentiation of bipolar cells in the rabbit retina.

    PubMed

    Wu, Mu-Ling; Chiao, Chuan-Chin

    2007-09-19

    Bipolar cells are responsible for transmitting light signals from the photoreceptors to the ganglion cells in the vertebrate retina. Their maturation process is not only important for establishing normal visual function, but may also underlie the dendritic remodeling of ganglion cells during development. It is known that light deprivation affects the synaptic connections of ganglion cells in the mammalian retina, but little is known about impact of visual experience on bipolar cell development. We used dye injection and gene gun labeling to identify bipolar cells, and characterized their morphological differentiation in normal-reared and dark-reared rabbits. Our results show that immature bipolar cells can be found as early as P1-3, and most characteristic bipolar cells can be identified during P4-6. More importantly, we found that light deprivation causes a delay rather than a permanent arrest of bipolar cell maturation in the rabbit retina. By eye opening at P10-11, both normal-reared and dark-reared rabbits possessed adult-like bipolar cells. This suggests that visual experience has a facilitating effect on the morphological differentiation of bipolar cells.

  5. Variable Cell Morphology Approach for Individual-Based Modeling of Microbial Communities

    PubMed Central

    Storck, Tomas; Picioreanu, Cristian; Virdis, Bernardino; Batstone, Damien J.

    2014-01-01

    An individual-based, mass-spring modeling framework has been developed to investigate the effect of cell properties on the structure of biofilms and microbial aggregates through Lagrangian modeling. Key features that distinguish this model are variable cell morphology described by a collection of particles connected by springs and a mechanical representation of deformable intracellular, intercellular, and cell-substratum links. A first case study describes the colony formation of a rod-shaped species on a planar substratum. This case shows the importance of mechanical interactions in a community of growing and dividing rod-shaped cells (i.e., bacilli). Cell-substratum links promote formation of mounds as opposed to single-layer biofilms, whereas filial links affect the roundness of the biofilm. A second case study describes the formation of flocs and development of external filaments in a mixed-culture activated sludge community. It is shown by modeling that distinct cell-cell links, microbial morphology, and growth kinetics can lead to excessive filamentous proliferation and interfloc bridging, possible causes for detrimental sludge bulking. This methodology has been extended to more advanced microbial morphologies such as filament branching and proves to be a very powerful tool in determining how fundamental controlling mechanisms determine diverse microbial colony architectures. PMID:24806936

  6. Supramolecular Approaches to Nanoscale Morphological Control in Organic Solar Cells

    PubMed Central

    Haruk, Alexander M.; Mativetsky, Jeffrey M.

    2015-01-01

    Having recently surpassed 10% efficiency, solar cells based on organic molecules are poised to become a viable low-cost clean energy source with the added advantages of mechanical flexibility and light weight. The best-performing organic solar cells rely on a nanostructured active layer morphology consisting of a complex organization of electron donating and electron accepting molecules. Although much progress has been made in designing new donor and acceptor molecules, rational control over active layer morphology remains a central challenge. Long-term device stability is another important consideration that needs to be addressed. This review highlights supramolecular strategies for generating highly stable nanostructured organic photovoltaic active materials by design. PMID:26110382

  7. A spectral and morphologic method for white blood cell classification

    NASA Astrophysics Data System (ADS)

    Wang, Qian; Chang, Li; Zhou, Mei; Li, Qingli; Liu, Hongying; Guo, Fangmin

    2016-10-01

    The identification of white blood cells is important as it provides an assay for diagnosis of various diseases. To overcome the complexity and inaccuracy of traditional methods based on light microscopy, we proposed a spectral and morphologic method based on hyperspectral blood images. We applied mathematical morphology-based methods to extract spatial information and supervised method is employed for spectral analysis. Experimental results show that white blood cells could be segmented and classified into five types with an overall accuracy of more than 90%. Moreover, the experiments including spectral features reached higher accuracy than the spatial-only cases, with a maximum improvement of nearly 20%. By combing both spatial and spectral features, the proposed method provides higher classification accuracy than traditional methods.

  8. Targeted cellular ablation based on the morphology of malignant cells.

    PubMed

    Ivey, Jill W; Latouche, Eduardo L; Sano, Michael B; Rossmeisl, John H; Davalos, Rafael V; Verbridge, Scott S

    2015-01-01

    Treatment of glioblastoma multiforme (GBM) is especially challenging due to a shortage of methods to preferentially target diffuse infiltrative cells, and therapy-resistant glioma stem cell populations. Here we report a physical treatment method based on electrical disruption of cells, whose action depends strongly on cellular morphology. Interestingly, numerical modeling suggests that while outer lipid bilayer disruption induced by long pulses (~100 μs) is enhanced for larger cells, short pulses (~1 μs) preferentially result in high fields within the cell interior, which scale in magnitude with nucleus size. Because enlarged nuclei represent a reliable indicator of malignancy, this suggested a means of preferentially targeting malignant cells. While we demonstrate killing of both normal and malignant cells using pulsed electric fields (PEFs) to treat spontaneous canine GBM, we proposed that properly tuned PEFs might provide targeted ablation based on nuclear size. Using 3D hydrogel models of normal and malignant brain tissues, which permit high-resolution interrogation during treatment testing, we confirmed that PEFs could be tuned to preferentially kill cancerous cells. Finally, we estimated the nuclear envelope electric potential disruption needed for cell death from PEFs. Our results may be useful in safely targeting the therapy-resistant cell niches that cause recurrence of GBM tumors. PMID:26596248

  9. Targeted cellular ablation based on the morphology of malignant cells

    PubMed Central

    Ivey, Jill W.; Latouche, Eduardo L.; Sano, Michael B.; Rossmeisl, John H.; Davalos, Rafael V.; Verbridge, Scott S.

    2015-01-01

    Treatment of glioblastoma multiforme (GBM) is especially challenging due to a shortage of methods to preferentially target diffuse infiltrative cells, and therapy-resistant glioma stem cell populations. Here we report a physical treatment method based on electrical disruption of cells, whose action depends strongly on cellular morphology. Interestingly, numerical modeling suggests that while outer lipid bilayer disruption induced by long pulses (~100 μs) is enhanced for larger cells, short pulses (~1 μs) preferentially result in high fields within the cell interior, which scale in magnitude with nucleus size. Because enlarged nuclei represent a reliable indicator of malignancy, this suggested a means of preferentially targeting malignant cells. While we demonstrate killing of both normal and malignant cells using pulsed electric fields (PEFs) to treat spontaneous canine GBM, we proposed that properly tuned PEFs might provide targeted ablation based on nuclear size. Using 3D hydrogel models of normal and malignant brain tissues, which permit high-resolution interrogation during treatment testing, we confirmed that PEFs could be tuned to preferentially kill cancerous cells. Finally, we estimated the nuclear envelope electric potential disruption needed for cell death from PEFs. Our results may be useful in safely targeting the therapy-resistant cell niches that cause recurrence of GBM tumors. PMID:26596248

  10. Targeted cellular ablation based on the morphology of malignant cells

    NASA Astrophysics Data System (ADS)

    Ivey, Jill W.; Latouche, Eduardo L.; Sano, Michael B.; Rossmeisl, John H.; Davalos, Rafael V.; Verbridge, Scott S.

    2015-11-01

    Treatment of glioblastoma multiforme (GBM) is especially challenging due to a shortage of methods to preferentially target diffuse infiltrative cells, and therapy-resistant glioma stem cell populations. Here we report a physical treatment method based on electrical disruption of cells, whose action depends strongly on cellular morphology. Interestingly, numerical modeling suggests that while outer lipid bilayer disruption induced by long pulses (~100 μs) is enhanced for larger cells, short pulses (~1 μs) preferentially result in high fields within the cell interior, which scale in magnitude with nucleus size. Because enlarged nuclei represent a reliable indicator of malignancy, this suggested a means of preferentially targeting malignant cells. While we demonstrate killing of both normal and malignant cells using pulsed electric fields (PEFs) to treat spontaneous canine GBM, we proposed that properly tuned PEFs might provide targeted ablation based on nuclear size. Using 3D hydrogel models of normal and malignant brain tissues, which permit high-resolution interrogation during treatment testing, we confirmed that PEFs could be tuned to preferentially kill cancerous cells. Finally, we estimated the nuclear envelope electric potential disruption needed for cell death from PEFs. Our results may be useful in safely targeting the therapy-resistant cell niches that cause recurrence of GBM tumors.

  11. Time Dependent Assessment of Morphological Changes: Leukodepleted Packed Red Blood Cells Stored in SAGM

    PubMed Central

    2016-01-01

    Usually packed red blood cells (pRBCs) require specific conditions in storage procedures to ensure the maximum shelf life of up to 42 days in 2–6°C. However, molecular and biochemical consequences can affect the stored blood cells; these changes are collectively labeled as storage lesions. In this study, the effect of prolonged storage was assessed through investigating morphological changes and evaluating oxidative stress. Samples from leukodepleted pRBC in SAGM stored at 4°C for 42 days were withdrawn aseptically on day 0, day 14, day 28, and day 42. Morphological changes were observed using scanning electron microscopy and correlated with osmotic fragility and hematocrit. Oxidative injury was studied through assessing MDA level as a marker for lipid peroxidation. Osmotic fragility test showed that extended storage time caused increase in the osmotic fragility. The hematocrit increased by 6.6% from day 0 to day 42. The last 2 weeks show alteration in the morphology with the appearance of echinocytes and spherocytes. Storage lesions and morphological alterations appeared to affect RBCs during the storage period. Further studies should be performed to develop strategies that will aid in the improvement of stored pRBC quality and efficacy. PMID:26904677

  12. ALS/FTLD-linked TDP-43 regulates neurite morphology and cell survival in differentiated neurons

    SciTech Connect

    Han, Jeong-Ho; Yu, Tae-Hoon; Ryu, Hyun-Hee; Jun, Mi-Hee; Ban, Byung-Kwan; Jang, Deok-Jin; Lee, Jin-A

    2013-08-01

    Tar-DNA binding protein of 43 kDa (TDP-43) has been characterized as a major component of protein aggregates in brains with neurodegenerative diseases such as frontotemporal lobar degeneration (FTLD) and amyotrophic lateral sclerosis (ALS). However, physiological roles of TDP-43 and early cellular pathogenic effects caused by disease associated mutations in differentiated neurons are still largely unknown. Here, we investigated the physiological roles of TDP-43 and the effects of missense mutations associated with diseases in differentiated cortical neurons. The reduction of TDP-43 by siRNA increased abnormal neurites and decreased cell viability. ALS/FTLD-associated missense mutant proteins (A315T, Q331K, and M337V) were partially mislocalized to the cytosol and neurites when compared to wild-type and showed abnormal neurites similar to those observed in cases of loss of TDP-43. Interestingly, cytosolic expression of wild-type TDP-43 with mutated nuclear localization signals also induced abnormal neurtie morphology and reduction of cell viability. However, there was no significant difference in the effects of cytosolic expression in neuronal morphology and cell toxicity between wild-type and missense mutant proteins. Thus, our results suggest that mislocalization of missense mutant TDP-43 may contribute to loss of TDP-43 function and affect neuronal morphology, probably via dominant negative action before severe neurodegeneration in differentiated cortical neurons. Highlights: • The function of nuclear TDP-43 in neurite morphology in mature neurons. • Partial mislocalization of TDP-43 missense mutants into cytosol from nucleus. • Abnormal neurite morphology caused by missense mutants of TDP-43. • The effect of cytosolic expression of TDP-43 in neurite morphology and in cell survival.

  13. Plasma cell morphology in multiple myeloma and related disorders.

    PubMed

    Ribourtout, B; Zandecki, M

    2015-06-01

    Normal and reactive plasma cells (PC) are easy to ascertain on human bone marrow films, due to their small mature-appearing nucleus and large cytoplasm, the latter usually deep blue after Giemsa staining. Cytoplasm is filled with long strands of rough endoplasmic reticulum and one large Golgi apparatus (paranuclear hof), demonstrating that PC are dedicated mainly to protein synthesis and excretion (immunoglobulin). Deregulation of the genome may induce clonal expansion of one PC that will lead to immunoglobulin overproduction and eventually to one among the so-called PC neoplasms. In multiple myeloma (MM), the number of PC is over 10% in most patients studied. Changes in the morphology of myeloma PC may be inconspicuous as compared to normal PC (30-50% patients). In other instances PC show one or several morphological changes. One is related to low amount of cytoplasm, defining lymphoplasmacytoid myeloma (10-15% patients). In other cases (40-50% patients), named immature myeloma cases, nuclear-cytoplasmic asynchrony is observed: presence of one nucleolus, finely dispersed chromatin and/or irregular nuclear contour contrast with a still large and blue (mature) cytoplasm. A peculiar morphological change, corresponding to the presence of very immature PC named plasmablasts, is observed in 10-15% cases. Several prognostic morphological classifications have been published, as mature myeloma is related to favorable outcome and immature myeloma, peculiarly plasmablastic myeloma, is related to dismal prognosis. However, such classifications are no longer included in current prognostic schemes. Changes related to the nucleus are very rare in monoclonal gammopathy of unknown significance (MGUS). In contrast, anomalies related to the cytoplasm of PC, including color (flaming cells), round inclusions (Mott cells, Russell bodies), Auer rod-like or crystalline inclusions, are reported in myeloma cases as well as in MGUS and at times in reactive disorders. They do not correspond

  14. Fractal morphology of Beta vulgaris L. cell suspension culture permeabilized with Triton X-100®

    NASA Astrophysics Data System (ADS)

    Arenas-Ocampo, M.; Alamilla-Beltrán, L.; Vanegas-Espinoza, P.; Camacho-Díaz, B.; Campos-Mendiola, R.; Gutiérrez-López, G.; Jiménez-Aparicio, A.

    2012-02-01

    In this work, morphology of Beta vulgaris L. cells permeabilized with 0.7mM of Triton X-100® was evaluated using digital image processing and concepts of fractal dimension (perimeter- area relations). Important morphometric changes were found when the contact-time with chemical agent was increased. The size of cells decreased, the cells lost the roundness and their shape was more sinuous; this behaviour was a result of a probable shrinkage caused by the excess of exposure with the permeabilization agent. Morphology of B. vulgaris cells after permeabilization, exhibited a fractal nature since the slope of the ratio of the logarithm of the perimeter vs logarithm of the area was higher than unit. Fractal geometry of the cell morphology was affected as a result of the exposure to Triton X-100®. Those changes can be attributed to the loss of turgor and structure of the cell wall.

  15. Quantification of Dynamic Morphological Drug Responses in 3D Organotypic Cell Cultures by Automated Image Analysis

    PubMed Central

    Härmä, Ville; Schukov, Hannu-Pekka; Happonen, Antti; Ahonen, Ilmari; Virtanen, Johannes; Siitari, Harri; Åkerfelt, Malin; Lötjönen, Jyrki; Nees, Matthias

    2014-01-01

    Glandular epithelial cells differentiate into complex multicellular or acinar structures, when embedded in three-dimensional (3D) extracellular matrix. The spectrum of different multicellular morphologies formed in 3D is a sensitive indicator for the differentiation potential of normal, non-transformed cells compared to different stages of malignant progression. In addition, single cells or cell aggregates may actively invade the matrix, utilizing epithelial, mesenchymal or mixed modes of motility. Dynamic phenotypic changes involved in 3D tumor cell invasion are sensitive to specific small-molecule inhibitors that target the actin cytoskeleton. We have used a panel of inhibitors to demonstrate the power of automated image analysis as a phenotypic or morphometric readout in cell-based assays. We introduce a streamlined stand-alone software solution that supports large-scale high-content screens, based on complex and organotypic cultures. AMIDA (Automated Morphometric Image Data Analysis) allows quantitative measurements of large numbers of images and structures, with a multitude of different spheroid shapes, sizes, and textures. AMIDA supports an automated workflow, and can be combined with quality control and statistical tools for data interpretation and visualization. We have used a representative panel of 12 prostate and breast cancer lines that display a broad spectrum of different spheroid morphologies and modes of invasion, challenged by a library of 19 direct or indirect modulators of the actin cytoskeleton which induce systematic changes in spheroid morphology and differentiation versus invasion. These results were independently validated by 2D proliferation, apoptosis and cell motility assays. We identified three drugs that primarily attenuated the invasion and formation of invasive processes in 3D, without affecting proliferation or apoptosis. Two of these compounds block Rac signalling, one affects cellular cAMP/cGMP accumulation. Our approach supports

  16. Case study of the morphologic variation of circulating tumor cells.

    PubMed

    Marrinucci, Dena; Bethel, Kelly; Bruce, Richard H; Curry, Douglas N; Hsieh, Ben; Humphrey, Mark; Krivacic, Robert T; Kroener, Joan; Kroener, Lindsay; Ladanyi, Andras; Lazarus, Nicole H; Nieva, Jorge; Kuhn, Peter

    2007-03-01

    We report a detailed cytomorphologic evaluation of the circulating component of widely metastatic breast carcinoma. A previously healthy 38-year-old woman was diagnosed with breast cancer. Wide local excision revealed a 1.7-cm infiltrating ductal adenocarcinoma, BSR score 7/9 with angiolymphatic invasion, and 4/20 lymph nodes positive for carcinoma. Five years later, a bone marrow biopsy revealed involvement of bone marrow by metastatic breast carcinoma, and shortly thereafter, metastases were identified in the liver and lung hilum. She enrolled in a clinical investigation for the detection of circulating tumor cells (CTCs) in breast carcinoma. A total of 659 CTCs were identified in a 10-mL blood sample using an immunofluorescent protocol targeting cytokeratins and detected using fiber-optic array scanning technology. The detected CTCs were subsequently stained with a Wright-Giemsa stain, and representative cells were evaluated in detail by light microscopy for morphologic evaluation. We find that the patient's CTCs exhibit a high degree of pleomorphism including CTCs with high and low nuclear-to-cytoplasmic ratios along with CTCs exhibiting early and late apoptotic changes. In addition, in comparison with her tumor cells in other sites, the full morphologic spectrum of cancer cells present in primary and metastatic tumor is also present in peripheral blood circulation. PMID:17188328

  17. Native human adipose stromal cells: localization, morphology and phenotype

    PubMed Central

    Maumus, M; Peyrafitte, J-A; D'Angelo, R; Fournier-Wirth, C; Bouloumié, A; Casteilla, L; Sengenès, C; Bourin, P

    2011-01-01

    Objectives: Beside having roles in energy homeostasis and endocrine modulation, adipose tissue (AT) is now considered a promising source of mesenchymal stromal cells (adipose-derived stromal cells or ASCs) for regenerative medicine. Despite numerous studies on cultured ASCs, native human ASCs are rarely investigated. Indeed, the phenotype of ASCs in their native state, their localization within AT and comparison with bone marrow-derived mesenchymal stromal cells (BM-MSCs) has been poorly investigated. Design: To address these issues, the stroma vascular fraction (SVF) of human AT was extracted and native cell subtypes were isolated by immunoselection to study their clonogenic potential in culture. Immunohistology on samples of human AT in combination with reconstruction of confocal sections were performed in order to localize ASCs. Results: Compared with BM-MNCs, all native ASCs were found in the CD34+ cell fraction of the AT-SVF. Native ASCs expressed classical mesenchymal markers described for BM-MSCs. Interestingly, CD34 expression decreased during ASC cell culture and was negatively correlated with cell proliferation rate. Immunohistological analysis revealed that native ASCs exhibited specific morphological features with protrusions. They were found scattered in AT stroma and did not express in vivo pericytic markers such as NG2, CD140b or alpha-smooth muscle actin, which appeared during the culture process. Finally, ASCs spontaneous commitment to adipocytic lineage was enhanced in AT from obese humans. Conclusions: The use of complementary methodological approaches to study native human ASCs revealed their immunophenotype, their specific morphology, their location within AT and their stemness. Furthermore, our data strongly suggest that human ASCs participate in adipogenesis during AT development. PMID:21266947

  18. Substrate properties affect collective cell motion

    NASA Astrophysics Data System (ADS)

    Pegoraro, Adrian; Guo, Ming; Ehrlicher, Allen; Weitz, David

    2013-03-01

    When cells move collectively, cooperative motion, which is characterized by long range correlations in cell movement, is necessary for migration. This collective cell motion is influenced by cell-cell interactions as well as by cell-substrate coupling. Furthermore, on soft substrates it is possible for cells to mechanically couple over long distances through the substrate itself. By changing the properties of the substrate, it is possible to decouple some of these contributions and better understand the role they play in collective cell motion. We vary both the substrate stiffness and adhesion protein concentration and find changes in the collective cell motion of the cells despite only small differences in total cell density and average cell size in the confluent layers. We test these changes on polyacrylamide and PDMS substrates as well as on structured substrates made of PDMS posts that prevent mechanical coupling through the substrate while still allowing stiffness to be varied.

  19. Morphology and growth of murine cell lines on model biomaterials.

    PubMed

    Godek, Marisha L; Duchsherer, Nichole L; McElwee, Quinn; Grainger, David W

    2004-01-01

    All biomaterial implants are assaulted by the host "foreign body" immune response. Understanding the complex, dynamic relationship between cells, biomaterials and milieu is an important first step towards controlling this reaction. Material surface chemistry dictates protein adsorption, and thus subsequent cell interactions. The cell-implant is a microenvironment involving 1) proteins that coat the surface and 2) cells that interact with these proteins. Macrophages and fibroblasts are two cell types that interact with proteins on biomaterials surfaces and play different related, but equally important, roles in biomaterials rejection and implant failure. Growth characteristics of four murine cell lines on model biomaterials surfaces were examined. Murine monocyte-macrophages (RAW 264.7 and J774A.1), murine macrophage (IC-21) and murine fibroblast (NIH 3T3) cell lines were tested to determine whether differences exist in adhesion, proliferation, differentiation, spreading, and fusion (macrophage lineages only) on these surfaces. Differences were observed in the ability of cells to adhere to and subsequently proliferate on polymer surfaces. (Monocyte-) macrophages grew well on all surfaces tested and growth rates were measured on three representative polymer biomaterials surfaces: tissue culture polystyrene (TCPS), polystyrene, and Teflon-AF. J774A.1 cultures grown on TCPS and treated with exogenous cytokines IL-4 and GM-CSF were observed to contain multinucleate cells with unusual morphologies. Thus, (monocyte-) macrophage cell lines were found to effectively attach to and interrogate each surface presented, with evidence of extensive spreading on Teflon-AF surfaces, particularly in the IC-21 cultures. The J774A.1 line was able to proliferate and/or differentiate to more specialized cell types (multinucleate/dendritic-like cells) in the presence of soluble chemokine cues. PMID:15133927

  20. Hyperthermia effects on the cytoskeleton and on cell morphology.

    PubMed

    Coakley, W T

    1987-01-01

    around 43 degrees C includes central retraction of membrane, loss of microvilli, concentration of organelles in a juxtanuclear position, rounding up of the cell, retention of contact with the substratum by processes which are sometimes beaded and blebbing of the cell membrane. The morphological effects of heat are compared here with those of cytochalasin, colcemid and a number of morphology modifying agents. Blebbing of membrane is a fairly general response of cells to stress. Proteins in blebs diffuse as if released from a lateral constraint. Moderate heating has been shown to cause cortical microfilament separation from the plasma membrane.(ABSTRACT TRUNCATED AT 400 WORDS) PMID:3332484

  1. Correlating the morphological and light scattering properties of biological cells

    NASA Astrophysics Data System (ADS)

    Moran, Marina

    The scattered light pattern from a biological cell is greatly influenced by the internal structure and optical properties of the cell. This research project examines the relationships between the morphological and scattering properties of biological cells through numerical simulations. The mains goals are: (1) to develop a procedure to analytically model biological cells, (2) to quantitatively study the effects of a range of cell characteristics on the features of the light scattering patterns, and (3) to classify cells based on the features of their light scattering patterns. A procedure to create an analytical cell model was developed which extracted structural information from the confocal microscopic images of cells and allowed for the alteration of the cell structure in a controlled and systematic way. The influence of cell surface roughness, nuclear size, and mitochondrial volume density, spatial distribution, size and shape on the light scattering patterns was studied through numerical simulations of light scattering using the Discrete Dipole Approximation. It was found that the light scattering intensity in the scattering angle range of 25° to 45° responded to changes in the surface fluctuation of the cell and the range of 90° to 110° was well suited for characterization of mitochondrial density and nuclear size. A comparison of light scattering pattern analysis methods revealed that the angular distribution of the scattered light and Gabor filters were most helpful in differentiating between the cell characteristics. In addition, a measured increase in the Gabor energy of the light scattering patterns in response to an increase in the complexity of the cell models suggested that a complex nuclear structure and mitochondria should be included when modeling biological cells for light scattering simulations. Analysis of the scattering pattern features with Gabor filters resulted in discrimination of the cell models according to cell surface roughness

  2. Calmodulin inhibition regulates morphological and functional changes related to the actin cytoskeleton in pure microglial cells.

    PubMed

    Szabo, Melinda; Dulka, Karolina; Gulya, Karoly

    2016-01-01

    The roles of calmodulin (CaM), a multifunctional intracellular calcium receptor protein, as concerns selected morphological and functional characteristics of pure microglial cells derived from mixed primary cultures from embryonal forebrains of rats, were investigated through use of the CaM antagonists calmidazolium (CALMID) and trifluoperazine (TFP). The intracellular localization of the CaM protein relative to phalloidin, a bicyclic heptapeptide that binds only to filamentous actin, and the ionized calcium-binding adaptor molecule 1 (Iba1), a microglia-specific actin-binding protein, was determined by immunocytochemistry, with quantitative analysis by immunoblotting. In unchallenged and untreated (control) microglia, high concentrations of CaM protein were found mainly perinuclearly in ameboid microglia, while the cell cortex had a smaller CaM content that diminished progressively deeper into the branches in the ramified microglia. The amounts and intracellular distributions of both Iba1 and CaM proteins were altered after lipopolysaccharide (LPS) challenge in activated microglia. CALMID and TFP exerted different, sometimes opposing, effects on many morphological, cytoskeletal and functional characteristics of the microglial cells. They affected the CaM and Iba1 protein expressions and their intracellular localizations differently, inhibited cell proliferation, viability and fluid-phase phagocytosis to different degrees both in unchallenged and in LPS-treated (immunologically challenged) cells, and differentially affected the reorganization of the actin cytoskeleton in the microglial cell cortex, influencing lamellipodia, filopodia and podosome formation. In summary, these CaM antagonists altered different aspects of filamentous actin-based cell morphology and related functions with variable efficacy, which could be important in deciphering the roles of CaM in regulating microglial functions in health and disease.

  3. Morphological homogeneity of neurons: searching for outlier neuronal cells.

    PubMed

    Zawadzki, Krissia; Feenders, Christoph; Viana, Matheus P; Kaiser, Marcus; Costa, Luciano da F

    2012-10-01

    We report a morphology-based approach for the automatic identification of outlier neurons, as well as its application to the NeuroMorpho.org database, with more than 5,000 neurons. Each neuron in a given analysis is represented by a feature vector composed of 20 measurements, which are then projected into a two-dimensional space by applying principal component analysis. Bivariate kernel density estimation is then used to obtain the probability distribution for the group of cells, so that the cells with highest probabilities are understood as archetypes while those with the smallest probabilities are classified as outliers. The potential of the methodology is illustrated in several cases involving uniform cell types as well as cell types for specific animal species. The results provide insights regarding the distribution of cells, yielding single and multi-variate clusters, and they suggest that outlier cells tend to be more planar and tortuous. The proposed methodology can be used in several situations involving one or more categories of cells, as well as for detection of new categories and possible artifacts. PMID:22615032

  4. WR-1065 and radioprotection of vascular endothelial cells. II. Morphology

    SciTech Connect

    Mooteri, S.N.; Podolski, J.L.; Drab, E.A.; Saclarides, T.J.

    1996-02-01

    Although the aminothiol WR-1065 protects normal tissues, its direct effect on the damage and restoration of the vascular endothelium is not clear. In endothelial cells, WR-1065 attenuates both the DNA damage and the G{sub 1}-phase arrest induced by radiation. After the destruction of nearby endothelial cells, the survivors rearrange their cytoskeleton, migrate and replicate. To determine the effect of radiation on morphology and migration, portions of bovine aortic endothelial cell cultures were denuded with a pipette tip and irradiated ({sup 137}Cs {gamma} rays). The following observations were noted after 5 Gy: within 10 min, there was increased formation of protein-mixed disulfides including actin-mixed disulfide; after 30 min, {alpha}{sub 5}{Beta}{sub 1}, the integrin receptor for fibronectin, was up-regulated on the apical membrane surface. Within 5 h, actin-containing stress fibers reorganized, although there was no change in the total filamentous (F-)actin content within the cells. Compared to controls after 24 h, the irradiated cells had migrated 15% farther (P < 0.01), and at the leading edge covered twice the surface area (P < 0.0001). The addition of 2 mM WR-1065 for 2 h before 5 Gy inhibited the increased expression of {alpha}{sub 5}{Beta}{sub 1}, promoted retention of stress fibers and prevented the enhanced cell migration and spreading. These results indicate that WR-1065 prevents radiation-induced morphological responses. This effect appears to be mediated by an impact on both adhesion molecule expression and cytoskeletal reorganization. 61 refs., 6 figs.

  5. Gene Therapy Restores Hair Cell Stereocilia Morphology in Inner Ears of Deaf Whirler Mice.

    PubMed

    Chien, Wade W; Isgrig, Kevin; Roy, Soumen; Belyantseva, Inna A; Drummond, Meghan C; May, Lindsey A; Fitzgerald, Tracy S; Friedman, Thomas B; Cunningham, Lisa L

    2016-02-01

    Hereditary deafness is one of the most common disabilities affecting newborns. Many forms of hereditary deafness are caused by morphological defects of the stereocilia bundles on the apical surfaces of inner ear hair cells, which are responsible for sound detection. We explored the effectiveness of gene therapy in restoring the hair cell stereocilia architecture in the whirlin mouse model of human deafness, which is deaf due to dysmorphic, short stereocilia. Wild-type whirlin cDNA was delivered via adeno-associated virus (AAV8) by injection through the round window of the cochleas in neonatal whirler mice. Subsequently, whirlin expression was detected in infected hair cells (IHCs), and normal stereocilia length and bundle architecture were restored. Whirlin gene therapy also increased inner hair cell survival in the treated ears compared to the contralateral nontreated ears. These results indicate that a form of inherited deafness due to structural defects in cochlear hair cells is amenable to restoration through gene therapy.

  6. Developmental mechanisms that regulate retinal ganglion cell dendritic morphology

    PubMed Central

    Tian, Ning

    2011-01-01

    One of the fundamental features of retinal ganglion cells (RGCs) is that dendrites of individual RGCs are confined to one or a few narrow strata within the inner plexiform layer (IPL), and each RGC synapses only with a small group of presynaptic bipolar and amacrine cells with axons/dendrites ramified in the same strata to process distinct visual features. The underlying mechanisms which control the development of this laminar-restricted distribution pattern of RGC dendrites have been extensively studied, and it is still an open question whether the dendritic pattern of RGCs is determined by molecular cues or by activity-dependent refinement. Accumulating evidence suggests that both molecular cues and activity-dependent refinement might regulate RGC dendrites in a cell subtype-specific manner. However, identification of morphological subtypes of RGCs before they have achieved their mature dendritic pattern is a major challenge in the study of RGC dendritic development. This problem is now being circumvented through the use of molecular markers in genetically engineered mouse lines to identify RGC subsets early during development. Another unanswered fundamental question in the study of activity-dependent refinement of RGC dendrites is how changes in synaptic activity lead to the changes in dendritic morphology. Recent studies have started to shed light on the molecular basis of activity-dependent dendritic refinement of RGCs by showing that some molecular cascades control the cytoskeleton reorganization of RGCs. PMID:21542137

  7. Morphological changes in neutron irradiated red blood cells.

    PubMed

    Nelson, A C; Wyle, H R

    1985-01-01

    Living human red blood cells (erythrocytes) were irradiated with a beam of thermal neutrons having a thermal neutron flux of 9.4 X 10(9) neutrons/cm2 per sec corresponding to a dose rate of 5 Gray per hour. The neutron beam was obtained from the thermal neutron facility at the MIT Nuclear Reactor and contained some gamma-ray contamination which contributes approximately 8% of the dose effect. Approximately 92% of the dose effect is due to the neutron radiation. Populations of neutron irradiated red blood cells were examined under scanning electron microscopy to observe morphological changes due to the radiation dose. The thermal neutron doses ranged from zero for controls to 75 Gray, and cell populations were examined at various post-irradiation time periods of 10, 48, and 96 h. A four-stage discoid to spheroid shape transformation of the damaged red blood cells was characterized, and the time dependence of each transformation stage was determined for both unirradiated and irradiated cells. The radiation dose caused an initial dose-dependent shift from Stage 1 to Stage 2 with an associated increase in the transformation rate constants. The thermal neutron doses delivered are considered to be in the low dose range for radiation effects on red blood cells, yet the pronounced effects indicate a high relative biological effectiveness (RBE) for thermal neutrons.

  8. Relationship of Treponema denticola periplasmic flagella to irregular cell morphology.

    PubMed Central

    Ruby, J D; Li, H; Kuramitsu, H; Norris, S J; Goldstein, S F; Buttle, K F; Charon, N W

    1997-01-01

    Treponema denticola is an anaerobic, motile, oral spirochete associated with periodontal disease. We found that the periplasmic flagella (PFs), which are located between the outer membrane sheath and cell cylinder, influence its morphology in a unique manner. In addition, the protein composition of the PFs was found to be quite complex and similar to those of other spirochetes. Dark-field microscopy revealed that most wild-type cells had an irregular twisted morphology, with both planar and helical regions, and a minority of cells had a regular right-handed helical shape. High-voltage electron microscopy indicated that the PFs, especially in those regions of the cell which were planar, wrapped around the cell body axis in a right-handed sense. In those regions of the cell which were helical or irregular, the PFs tended to lie along the cell axis. The PFs caused the cell to form the irregular shape, as two nonmotile, PF-deficient mutants (JR1 and HL51) were no longer irregular but were right-handed helices. JR1 was isolated as a spontaneously occurring nonmotile mutant, and HL51 was isolated as a site-directed mutant in the flagellar hook gene flgE. Consistent with these results is the finding that wild-type cells with their outer membrane sheath removed were also right-handed helices similar in shape to JR1 and HL51. Purified PFs were analyzed by two-dimensional gel electrophoresis, and several protein species were identified. Western blot analysis using antisera to Treponema pallidum PF proteins along with N-terminal amino acid sequence analysis indicated T. denticola PFs are composed of one class A sheath protein of 38 kDa (FlaA) and three class B proteins of 35 kDa (FlaB1 and FlaB2) and one of 34 kDa (FlaB3). The N-terminal amino acid sequences of the FlaA and FlaB proteins of T. denticola were most similar to those of T. pallidum and Treponema phagedenis. Because these proteins were present in markedly reduced amounts or were absent in HL51, PF synthesis is

  9. The sertolian epithelium in the testis of men affected by 'Sertoli-cell-only syndrome'.

    PubMed

    Tedde, G; Montella, A; Fiocca, D; Delrio, A N

    1993-01-01

    Because of the architectural complexity of the seminiferous epithelium, the Sertoli cell is extremely difficult to study. The individual cellular constituents of the tubular wall are intimately associated with one another; especially Sertoli cells and germinal cells are tightly connected. As implied by the name, Sertoli-cell-only syndrome (SCOS) is characterized by the presence of only Sertoli cells in the seminiferous tubule. The absence of germinal cells makes this condition ideal for the morphological study of Sertoli cell. Testicular biopsy specimens of subjects affected by SCOS were studied under light and electron microscopy. The Sertoli cells appeared to be morphologically normal, except for their shape, that appears to be columnar as result of the complete absence of the germinal cells. The cellular outlines were irregular, particularly at the base, but the cytoplasm contained normal organelles and inclusions. The presence of both pale and dark elements was evident. These differences in staining reflect the variability in concentration of glycogen particles and intermediate microfilaments in the cytoplasm. In spite of these differences between Sertoli cells in SCOS and those in normal subjects, SCOS represents a satisfactory model for the morphological and functional analysis of the Sertoli cells. PMID:7694556

  10. Heme-oxygenase-1 implications in cell morphology and the adhesive behavior of prostate cancer cells

    PubMed Central

    Gueron, Geraldine; Giudice, Jimena; Valacco, Pia; Paez, Alejandra; Elguero, Belen; Toscani, Martin; Jaworski, Felipe; Leskow, Federico Coluccio; Cotignola, Javier; Marti, Marcelo; Binaghi, Maria; Navone, Nora; Vazquez, Elba

    2014-01-01

    Prostate cancer (PCa) is the second leading cause of cancer death in men. Although previous studies in PCa have focused on cell adherens junctions (AJs), key players in metastasis, they have left the molecular mechanisms unexplored. Inflammation and the involvement of reactive oxygen species (ROS) are critical in the regulation of cell adhesion and the integrity of the epithelium. Heme oxygenase-1 (HO-1) counteracts oxidative and inflammatory damage. Here, we investigated whether HO-1 is implicated in the adhesive and morphological properties of tumor cells. Genes differentially regulated by HO-1 were enriched for cell motility and adhesion biological processes. HO-1 induction, increased E-cadherin and β-catenin levels. Immunofluorescence analyses showed a striking remodeling of E-cadherin/β-catenin based AJs under HO-1 modulation. Interestingly, the enhanced levels of E-cadherin and β-catenin coincided with a markedly change in cell morphology. To further our analysis we sought to identify HO-1 binding proteins that might participate in the regulation of cell morphology. A proteomics approach identified Muskelin, as a novel HO-1 partner, strongly implicated in cell morphology regulation. These results define a novel role for HO-1 in modulating the architecture of cell-cell interactions, favoring a less aggressive phenotype and further supporting its anti-tumoral function in PCa. PMID:24961479

  11. Jasmonic acid affects plant morphology and calcium-dependent protein kinase expression and activity in Solanum tuberosum.

    PubMed

    Ulloa, Rita M; Raíces, Marcela; MacIntosh, Gustavo C; Maldonado, Sara; Téllez-Iñón, María T

    2002-07-01

    The effect of jasmonic acid (JA) on plant growth and on calcium-dependent protein kinase (CDPK) activity and expression was studied in non-photoperiodic potato plants, Solanum tuberosum L. var. Spunta, grown in vitro. Stem cuttings were grown for 45 days (long treatment, LT) in MS medium with increasing concentrations of JA. For short treatments (ST) adult plants grown in MS were transferred for 1, 4 and 20 h to JA containing media. During the LT, low concentrations of JA promoted cell expansion and shoot elongation while higher concentrations caused growth inhibition. Under these conditions, treated plants showed root shortening and tuber formation was not induced. Morphological and histochemical studies using light microscopy and TEM analysis of leaves from treated plants revealed that JA also affected subcellular organelles of mesophyll cells. Peroxisomes increased in size and number, and an autophagic process was triggered in response to high concentrations of the hormone. CDPK activity, determined in crude extracts of treated plants (LT), was inhibited (up to 80%). Plant growth and CDPK inhibition were reverted upon transfer of the plants to hormone-free medium. Soluble CDPK activity decreased in response to JA short treatment. Concomitantly, a decline in the steady state levels of StCDPK2 mRNA, a potato CDPK isoform that is expressed in leaves, was observed. These data suggest that the phytohormone down-regulated the expression and activity of the kinase.

  12. Testicular structure and germ cells morphology in salamanders

    PubMed Central

    Uribe, Mari Carmen; Mejía-Roa, Víctor

    2014-01-01

    Testes of salamanders or urodeles are paired elongated organs that are attached to the dorsal wall of the body by a mesorchium. The testes are composed of one or several lobes. Each lobe is morphologically and functionally a similar testicular unit. The lobes of the testis are joined by cords covered by a single peritoneal epithelium and subjacent connective tissue. The cords contain spermatogonia. Spermatogonia associate with Sertoli cells to form spermatocysts or cysts. The spermatogenic cells in a cyst undergo their development through spermatogenesis synchronously. The distribution of cysts displays the cephalo-caudal gradient in respect to the stage of spermatogenesis. The formation of cysts at cephalic end of the testis causes their migration along the lobules to the caudal end. Consequently, the disposition in cephalo-caudal regions of spermatogenesis can be observed in longitudinal sections of the testis. The germ cells are spermatogonia, diploid cells with mitotic activity; primary and second spermatocytes characterized by meiotic divisions that develop haploid spermatids; during spermiogenesis the spermatids differentiate to spermatozoa. During spermiation the cysts open and spermatozoa leave the testicular lobules. After spermiation occurs the development of Leydig cells into glandular tissue. This glandular tissue regressed at the end of the reproductive cycle. PMID:26413406

  13. Pit cells as liver-associated natural killer cells: morphology and function.

    PubMed

    Nakatani, Kazuki; Kaneda, Kenji; Seki, Shuichi; Nakajima, Yuji

    2004-03-01

    Pit cells are one type of hepatic sinusoidal cells, defined morphologically as large granular lymphocytes (LGLs) and functionally as liver-associated natural killer (NK) cells. They are situated inside the sinusoidal lumen, adhering to the endothelial cells and Kupffer cells. They contain multivesicular body-related dense granules and rod-cored vesicles. The number and size of granules and vesicles differ between hepatic NK cells and peripheral blood cells, suggesting possible differentiation of the latter into the former in the microenvironment of the liver. In addition to NK cells, natural killer T (NKT) cells are also abundant in the liver. They share several morphological properties with NK cells, and at least some are probably observed as pit cells under an electron microscope. NK cells recognize target cells with their surface receptors such as inhibitory and activating receptors. They exert antitumor functions by exocytosis of perforin/granzyme-containing granules, induction of death receptor-mediated apoptosis in target cells, and production of various cytokines that augment the activities of other immune cells. NKT cells play important roles in initiating and assisting the function of NK cells by producing interferon-gamma.

  14. The renal tumor morphological characteristics that affect surgical planning for laparoscopic or open partial nephrectomy.

    PubMed

    Funahashi, Yasuhito; Murotani, Kenta; Yoshino, Yasushi; Sassa, Naoto; Ishida, Shohei; Gotoh, Momokazu

    2015-02-01

    The purpose of this study is to investigate the morphological characteristics of renal tumors which affect the surgeons' decision-making for the selection of open or laparoscopic partial nephrectomy. We included 147 patients who underwent partial nephrectomy for renal masses with elective indications in this study. Laparoscopic partial nephrectomy (LPN) and open partial nephrectomy (OPN) were performed in 72 and 75 patients, respectively. Preoperative trans-sectional images were used to assess tumor characteristics such as tumor size, endophyticity, distance from the sinus, distance from the kidney equator, hilar designation, inside designation, and R.E.N.A.L. nephrometry score. Univariate logistic regression analyses demonstrated that tumor size, endophyticity, distance from the sinus, hilar designation, inside designation, and R.E.N.A.L. nephrometry score were associated with decision of laparoscopic partial nephrectomy. Among these factors, multiple regression analyses showed that endophyticity (odds ratio = 0.92, p = 0.007) and distance from the sinus (odds ratio = 1.201, p < 0.001) had statistically significant associations with the type of operation performed. ROC analyses demonstrated cut-off values of 16 mm for endophyticity (sensitivity 69%, specificity 77%) and of 4 mm for distance from the sinus (sensitivity 79%, specificity 65%) for predicting the selection of laparoscopic surgery. In conclusion, this study revealed that endophyticity and distance from the sinus were important for the surgical planning of partial nephrectomy.

  15. A genetic screen for zygotic embryonic lethal mutations affecting cuticular morphology in the wasp Nasonia vitripennis.

    PubMed Central

    Pultz, M A; Zimmerman, K K; Alto, N M; Kaeberlein, M; Lange, S K; Pitt, J N; Reeves, N L; Zehrung, D L

    2000-01-01

    We have screened for zygotic embryonic lethal mutations affecting cuticular morphology in Nasonia vitripennis (Hymenoptera; Chalcidoidea). Our broad goal was to investigate the use of Nasonia for genetically surveying conservation and change in regulatory gene systems, as a means to understand the diversity of developmental strategies that have arisen during the course of evolution. Specifically, we aim to compare anteroposterior patterning gene functions in two long germ band insects, Nasonia and Drosophila. In Nasonia, unfertilized eggs develop as haploid males while fertilized eggs develop as diploid females, so the entire genome can be screened for recessive zygotic mutations by examining the progeny of F1 females. We describe 74 of >100 lines with embryonic cuticular mutant phenotypes, including representatives of coordinate, gap, pair-rule, segment polarity, homeotic, and Polycomb group functions, as well as mutants with novel phenotypes not directly comparable to those of known Drosophila genes. We conclude that Nasonia is a tractable experimental organism for comparative developmental genetic study. The mutants isolated here have begun to outline the extent of conservation and change in the genetic programs controlling embryonic patterning in Nasonia and Drosophila. PMID:10866651

  16. Xp11 translocation renal cell carcinoma morphologically mimicking clear cell-papillary renal cell carcinoma in an adult patient: report of a case expanding the morphologic spectrum of Xp11 translocation renal cell carcinomas.

    PubMed

    Parihar, Asmita; Tickoo, Satish K; Kumar, Sunil; Arora, Vinod Kumar

    2015-05-01

    Xp11 translocation renal cell carcinoma (RCC) is a relatively rare tumor mainly affecting children and adolescents. It shows significant morphological overlap with the 2 most common adult renal tumors, which are the clear cell (conventional) RCC and papillary RCC. We describe case of a young adult female who presented with right flank pain and abdominal mass. Radiological investigations showed features suggestive of renal cell carcinoma in the right kidney. Histopathological findings while suggestive of Xp11 carcinoma, showed significant overlap with the recently described entity clear cell papillary RCC. TFE3 immunohistochemistry confirmed the tumor to be Xp11 translocation RCC. The patient had an aggressive course with lymph node metastasis. In this report, we discuss differential diagnosis and the diagnostic challenges of Xp11 translocation RCC in adults.

  17. Alteration of Cell Morphology with Nano- and Micro-Topographical Surface on Closed-Packed Silica Nanobeads.

    PubMed

    Park, Jeong Su; Lee, Dahyun; Park, Ji Yeon; Lee, Hyo Jin; Kim, Jin-Seok; Lee, Jin Seok

    2016-06-01

    The nanotopological cues are emerging field of in vivo study, because it stimulates alteration of cell morphology and cell behavior such as adhesion, proliferation, differentiation, apoptosis and migration. However, it has not studies nanosurface affected cancer cell behavior, therefore, in this study, we determined that effects the silica nanobeads used as nanotopological tools on cancer cells. For synthesis of silica beads, we made it using stober method and basic amino acid (L-arginine) instead of NH4OH. We carried out rubbing beads to obtain the monolayer silica beads and it used as nanotopological cues for fabrication. It was induced changing cell morphology at 1DIV in group-II (SB-450 and SB-570). However, it was maintained in group-I (SB-118, SB-230) and group-III (SB-1450) like control. Therefore, we separated type-I and type-II surface along with area of cell adhesion and morphology. The characteristic of type-II surface was long distance between contact points, resulting in increase of tension to cells. We found that the morphology rounded up by type-II surface at 1DIV. We described that the nanosurface-induced mechanical tension is associated with alteration of morphology, thus the silica nanobeads used as nanotopological tools for cancer research.

  18. Alteration of Cell Morphology with Nano- and Micro-Topographical Surface on Closed-Packed Silica Nanobeads.

    PubMed

    Park, Jeong Su; Lee, Dahyun; Park, Ji Yeon; Lee, Hyo Jin; Kim, Jin-Seok; Lee, Jin Seok

    2016-06-01

    The nanotopological cues are emerging field of in vivo study, because it stimulates alteration of cell morphology and cell behavior such as adhesion, proliferation, differentiation, apoptosis and migration. However, it has not studies nanosurface affected cancer cell behavior, therefore, in this study, we determined that effects the silica nanobeads used as nanotopological tools on cancer cells. For synthesis of silica beads, we made it using stober method and basic amino acid (L-arginine) instead of NH4OH. We carried out rubbing beads to obtain the monolayer silica beads and it used as nanotopological cues for fabrication. It was induced changing cell morphology at 1DIV in group-II (SB-450 and SB-570). However, it was maintained in group-I (SB-118, SB-230) and group-III (SB-1450) like control. Therefore, we separated type-I and type-II surface along with area of cell adhesion and morphology. The characteristic of type-II surface was long distance between contact points, resulting in increase of tension to cells. We found that the morphology rounded up by type-II surface at 1DIV. We described that the nanosurface-induced mechanical tension is associated with alteration of morphology, thus the silica nanobeads used as nanotopological tools for cancer research. PMID:27427740

  19. Neuronize: a tool for building realistic neuronal cell morphologies

    PubMed Central

    Brito, Juan P.; Mata, Susana; Bayona, Sofia; Pastor, Luis; DeFelipe, Javier; Benavides-Piccione, Ruth

    2013-01-01

    This study presents a tool, Neuronize, for building realistic three-dimensional models of neuronal cells from the morphological information extracted through computer-aided tracing applications. Neuronize consists of a set of methods designed to build 3D neural meshes that approximate the cell membrane at different resolution levels, allowing a balance to be reached between the complexity and the quality of the final model. The main contribution of the present study is the proposal of a novel approach to build a realistic and accurate 3D shape of the soma from the incomplete information stored in the digitally traced neuron, which usually consists of a 2D cell body contour. This technique is based on the deformation of an initial shape driven by the position and thickness of the first order dendrites. The addition of a set of spines along the dendrites completes the model, building a final 3D neuronal cell suitable for its visualization in a wide range of 3D environments. PMID:23761740

  20. Cell wall staining with Trypan blue enables quantitative analysis of morphological changes in yeast cells

    PubMed Central

    Liesche, Johannes; Marek, Magdalena; Günther-Pomorski, Thomas

    2015-01-01

    Yeast cells are protected by a cell wall that plays an important role in the exchange of substances with the environment. The cell wall structure is dynamic and can adapt to different physiological states or environmental conditions. For the investigation of morphological changes, selective staining with fluorescent dyes is a valuable tool. Furthermore, cell wall staining is used to facilitate sub-cellular localization experiments with fluorescently-labeled proteins and the detection of yeast cells in non-fungal host tissues. Here, we report staining of Saccharomyces cerevisiae cell wall with Trypan Blue, which emits strong red fluorescence upon binding to chitin and yeast glucan; thereby, it facilitates cell wall analysis by confocal and super-resolution microscopy. The staining pattern of Trypan Blue was similar to that of the widely used UV-excitable, blue fluorescent cell wall stain Calcofluor White. Trypan Blue staining facilitated quantification of cell size and cell wall volume when utilizing the optical sectioning capacity of a confocal microscope. This enabled the quantification of morphological changes during growth under anaerobic conditions and in the presence of chemicals, demonstrating the potential of this approach for morphological investigations or screening assays. PMID:25717323

  1. Cell wall staining with Trypan blue enables quantitative analysis of morphological changes in yeast cells.

    PubMed

    Liesche, Johannes; Marek, Magdalena; Günther-Pomorski, Thomas

    2015-01-01

    Yeast cells are protected by a cell wall that plays an important role in the exchange of substances with the environment. The cell wall structure is dynamic and can adapt to different physiological states or environmental conditions. For the investigation of morphological changes, selective staining with fluorescent dyes is a valuable tool. Furthermore, cell wall staining is used to facilitate sub-cellular localization experiments with fluorescently-labeled proteins and the detection of yeast cells in non-fungal host tissues. Here, we report staining of Saccharomyces cerevisiae cell wall with Trypan Blue, which emits strong red fluorescence upon binding to chitin and yeast glucan; thereby, it facilitates cell wall analysis by confocal and super-resolution microscopy. The staining pattern of Trypan Blue was similar to that of the widely used UV-excitable, blue fluorescent cell wall stain Calcofluor White. Trypan Blue staining facilitated quantification of cell size and cell wall volume when utilizing the optical sectioning capacity of a confocal microscope. This enabled the quantification of morphological changes during growth under anaerobic conditions and in the presence of chemicals, demonstrating the potential of this approach for morphological investigations or screening assays.

  2. Fructose affecting morphology and inducing β-fructofuranosidases in Penicillium janczewskii.

    PubMed

    Pessoni, Rosemeire A B; Tersarotto, Carla C; Mateus, Cássia A P; Zerlin, Juliana K; Simões, Kelly; de Cássia L Figueiredo-Ribeiro, Rita; Braga, Márcia R

    2015-01-01

    Fructose, glucose, and an equimolar mixture of both sugars affected differently hyphae thickness, biomass production and secretion of β-fructofuranosidase in Penicillium janczewskii. Reduced growth, thinner hyphae and visible injuries were early observed during fungal cultivation in fructose-containing medium, reaching the maximum between 12 and 15 days of culture. Total sugar content from the cell wall was lower when fructose was supplied and polysaccharides lower than 10 kDa predominated, regardless the culture age. Maximal inulinase and invertase activities were detected in culture filtrates after 12 days, excepting in the glucose-containing medium. Structural changes in cell walls coincided with the increase of extracellular enzyme activity in the fructose-containing medium. The fragility of the hyphae might be related with both low carbohydrate content and predominance of low molecular weight glucans in the walls. Data presented here suggest changes in carbohydrate component of the cell walls are induced by the carbon source. PMID:26380163

  3. [The morphological and karyological characteristics of MDCK and vero (B) cells cultures on plant hydrolyzate-based nutrient media].

    PubMed

    Mikhailova, G R; Mazurkova, N A; Podchernyaeva, R Ya; Ryabchikova, E I; Troshkova, G P; Shishkina, L N

    2011-01-01

    MDCK and Vero (B) cell cultures were propagated during 10 passages in the experimental nutrient media containing the soybean powder hydrolyzate prepared using trypsin and bromelain enzymes and the rice powder hydrolysate prepared with trypsin and in the control DMEM and SFM4 MegaVir media. The karyological, morphological, and proliferative characteristics of continuous cultures were examined and compared. The experimental media supplied with 3% fetal bovine serum (FBS) (Gibco, U.S.A.) showed high growth-enhancing properties and failed to affect their morphology. After propagated during 10 passages in the experimental media preserved a stable karyotype. MDCK cell cultures in the nutrient media based on rice and soybean powder hydrolyzates low (2%) in FBS caused no substantial changes in the proliferation indices and morphological and karyological characteristics of cell cultures.

  4. Bisphenol A affects placental layers morphology and angiogenesis during early pregnancy phase in mice.

    PubMed

    Tait, Sabrina; Tassinari, Roberta; Maranghi, Francesca; Mantovani, Alberto

    2015-11-01

    Bisphenol A (BPA) is a widespread endocrine disrupter mainly used in food contact plastics. Much evidence supports the adverse effects of BPA, particularly on susceptible groups such as pregnant women. The present study considered placental development - relevant for pregnancy outcomes and fetal nutrition/programming - as a potential target of BPA. Pregnant CD-1 mice were administered per os with vehicle, 0.5 (BPA05) or 50 mg kg(-1) (BPA50) body weight day(-1) of BPA, from gestational day (GD) 1 to GD11. At GD12, BPA50 induced significant degeneration and necrosis of giant cells, increased vacuolization in the junctional zone in the absence of glycogen accumulation and reduction of the spongiotrophoblast layer. In addition, BPA05 induced glycogen depletion as well as significant nuclear accumulation of β-catenin in trophoblasts of labyrinthine and spongiotrophoblast layers, supporting the activation of the Wnt/β-catenin pathway. Transcriptomic analysis indicated that BPA05 promoted and BPA50 inhibited blood vessel development and branching; morphologically, maternal vessels were narrower in BPA05 placentas, whereas embryonic and maternal vessels were irregularly dilated in the labyrinth of BPA50 placentas. Quantitative polymerase chain reaction evidenced an estrogen receptor β induction by BPA50, which did not correspond to downstream genes activation; indeed, the transcription factor binding sites analysis supported the AhR/Arnt complex as regulator of BPA50-modulated genes. Conversely, Creb appeared as the main transcription factor regulating BPA05-modulated genes. Embryonic structures (head, forelimb) showed divergent perturbations upon BPA05 or BPA50 exposure, potentially related to unbalanced embryonic nutrition and/or to modulation of genes involved in embryo development. Our findings support placenta as an important target of BPA, even at environmentally relevant dose levels. PMID:26063408

  5. The Morphological and Molecular Changes of Brain Cells Exposed to Direct Current Electric Field Stimulation

    PubMed Central

    Pelletier, Simon J.; Lagacé, Marie; St-Amour, Isabelle; Arsenault, Dany; Cisbani, Giulia; Chabrat, Audrey; Fecteau, Shirley; Lévesque, Martin

    2015-01-01

    Background: The application of low-intensity direct current electric fields has been experimentally used in the clinic to treat a number of brain disorders, predominantly using transcranial direct current stimulation approaches. However, the cellular and molecular changes induced by such treatment remain largely unknown. Methods: Here, we tested various intensities of direct current electric fields (0, 25, 50, and 100V/m) in a well-controlled in vitro environment in order to investigate the responses of neurons, microglia, and astrocytes to this type of stimulation. This included morphological assessments of the cells, viability, as well as shape and fiber outgrowth relative to the orientation of the direct current electric field. We also undertook enzyme-linked immunosorbent assays and western immunoblotting to identify which molecular pathways were affected by direct current electric fields. Results: In response to direct current electric field, neurons developed an elongated cell body shape with neurite outgrowth that was associated with a significant increase in growth associated protein-43. Fetal midbrain dopaminergic explants grown in a collagen gel matrix also showed a reorientation of their neurites towards the cathode. BV2 microglial cells adopted distinct morphological changes with an increase in cyclooxygenase-2 expression, but these were dependent on whether they had already been activated with lipopolysaccharide. Finally, astrocytes displayed elongated cell bodies with cellular filopodia that were oriented perpendicularly to the direct current electric field. Conclusion: We show that cells of the central nervous system can respond to direct current electric fields both in terms of their morphological shape and molecular expression of certain proteins, and this in turn can help us to begin understand the mechanisms underlying the clinical benefits of direct current electric field. PMID:25522422

  6. The Influence of Genome and Cell Size on Brain Morphology in Amphibians.

    PubMed

    Roth, Gerhard; Walkowiak, Wolfgang

    2015-08-10

    In amphibians, nerve cell size is highly correlated with genome size, and increases in genome and cell size cause a retardation of the rate of development of nervous (as well as nonnervous) tissue leading to secondary simplification. This yields an inverse relationship between genome and cell size on the one hand and morphological complexity of the tectum mesencephali as the main visual center, the size of the torus semicircularis as the main auditory center, the size of the amphibian papilla as an important peripheral auditory structure, and the size of the cerebellum as a major sensorimotor center. Nervous structures developing later (e.g., torus and cerebellum) are more affected by secondary simplification than those that develop earlier (e.g., the tectum). This effect is more prominent in salamanders and caecilians than in frogs owing to larger genome and cells sizes in the former two taxa. We hypothesize that because of intragenomic evolutionary processes, important differences in brain morphology can arise independently of specific environmental selection.

  7. The Influence of Genome and Cell Size on Brain Morphology in Amphibians.

    PubMed

    Roth, Gerhard; Walkowiak, Wolfgang

    2015-09-01

    In amphibians, nerve cell size is highly correlated with genome size, and increases in genome and cell size cause a retardation of the rate of development of nervous (as well as nonnervous) tissue leading to secondary simplification. This yields an inverse relationship between genome and cell size on the one hand and morphological complexity of the tectum mesencephali as the main visual center, the size of the torus semicircularis as the main auditory center, the size of the amphibian papilla as an important peripheral auditory structure, and the size of the cerebellum as a major sensorimotor center. Nervous structures developing later (e.g., torus and cerebellum) are more affected by secondary simplification than those that develop earlier (e.g., the tectum). This effect is more prominent in salamanders and caecilians than in frogs owing to larger genome and cells sizes in the former two taxa. We hypothesize that because of intragenomic evolutionary processes, important differences in brain morphology can arise independently of specific environmental selection. PMID:26261281

  8. How does cancer cell metabolism affect tumor migration and invasion?

    PubMed

    Han, Tianyu; Kang, De; Ji, Daokun; Wang, Xiaoyu; Zhan, Weihua; Fu, Minggui; Xin, Hong-Bo; Wang, Jian-Bin

    2013-01-01

    Cancer metastasis is the major cause of cancer-associated death. Accordingly, identification of the regulatory mechanisms that control whether or not tumor cells become "directed walkers" is a crucial issue of cancer research. The deregulation of cell migration during cancer progression determines the capacity of tumor cells to escape from the primary tumors and invade adjacent tissues to finally form metastases. The ability to switch from a predominantly oxidative metabolism to glycolysis and the production of lactate even when oxygen is plentiful is a key characteristic of cancer cells. This metabolic switch, known as the Warburg effect, was first described in 1920s, and affected not only tumor cell growth but also tumor cell migration. In this review, we will focus on the recent studies on how cancer cell metabolism affects tumor cell migration and invasion. Understanding the new aspects on molecular mechanisms and signaling pathways controlling tumor cell migration is critical for development of therapeutic strategies for cancer patients.

  9. Alpha-Synuclein affects neurite morphology, autophagy, vesicle transport and axonal degeneration in CNS neurons

    PubMed Central

    Koch, J C; Bitow, F; Haack, J; d'Hedouville, Z; Zhang, J-N; Tönges, L; Michel, U; Oliveira, L M A; Jovin, T M; Liman, J; Tatenhorst, L; Bähr, M; Lingor, P

    2015-01-01

    Many neuropathological and experimental studies suggest that the degeneration of dopaminergic terminals and axons precedes the demise of dopaminergic neurons in the substantia nigra, which finally results in the clinical symptoms of Parkinson disease (PD). The mechanisms underlying this early axonal degeneration are, however, still poorly understood. Here, we examined the effects of overexpression of human wildtype alpha-synuclein (αSyn-WT), a protein associated with PD, and its mutant variants αSyn-A30P and -A53T on neurite morphology and functional parameters in rat primary midbrain neurons (PMN). Moreover, axonal degeneration after overexpression of αSyn-WT and -A30P was analyzed by live imaging in the rat optic nerve in vivo. We found that overexpression of αSyn-WT and of its mutants A30P and A53T impaired neurite outgrowth of PMN and affected neurite branching assessed by Sholl analysis in a variant-dependent manner. Surprisingly, the number of primary neurites per neuron was increased in neurons transfected with αSyn. Axonal vesicle transport was examined by live imaging of PMN co-transfected with EGFP-labeled synaptophysin. Overexpression of all αSyn variants significantly decreased the number of motile vesicles and decelerated vesicle transport compared with control. Macroautophagic flux in PMN was enhanced by αSyn-WT and -A53T but not by αSyn-A30P. Correspondingly, colocalization of αSyn and the autophagy marker LC3 was reduced for αSyn-A30P compared with the other αSyn variants. The number of mitochondria colocalizing with LC3 as a marker for mitophagy did not differ among the groups. In the rat optic nerve, both αSyn-WT and -A30P accelerated kinetics of acute axonal degeneration following crush lesion as analyzed by in vivo live imaging. We conclude that αSyn overexpression impairs neurite outgrowth and augments axonal degeneration, whereas axonal vesicle transport and autophagy are severely altered. PMID:26158517

  10. Can neurodegenerative disease be defined by four 'primary determinants': anatomy, cells, molecules, and morphology?

    PubMed

    Armstrong, R A

    2016-01-01

    Traditional methods of describing and classifying neurodegenerative disease are based on the clinico-pathological concept supported by molecular pathological studies and defined by 'consensus criteria'. Disease heterogeneity, overlap between disorders, and the presence of multiple co-pathologies, however, have questioned the validity and status of many traditional disorders. If cases of neurodegenerative disease are not easily classifiable into distinct entities, but more continuously distributed, then a new descriptive framework may be required. This review proposes that there are four key neuropathological features of neurodegenerative disease (the 'primary determinants') that could be used to provide such a framework, viz., the anatomical pathways affected by the disease ('anatomy'), the cell populations affected ('cells'), the molecular pathology of 'signature' pathological lesions ('molecules'), and the morphological types of neurodegeneration ('morphology'). This review first discusses the limitations of existing classificatory systems and second provides evidence that the four primary determinants could be used as axes to define all cases of neurodegenerative disease. To illustrate the methodology, the primary determinants were applied to the study of a group of closely related tauopathy cases and to heterogeneity within frontotemporal lobar degeneration with TDP-43 proteinopathy (FTLD-TDP). PMID:27543767

  11. Modulation of Hepatocarcinoma Cell Morphology and Activity by Parylene-C Coating on PDMS

    PubMed Central

    Guimard, Denis; Arakawa, Yasuhiko; Sakai, Yasuyuki; Fujii, Teruo

    2010-01-01

    Background The ability to understand and locally control the morphogenesis of mammalian cells is a fundamental objective of cell and developmental biology as well as tissue engineering research. We present parylene-C (ParC) deposited on polydimethylsiloxane (PDMS) as a new substratum for in vitro advanced cell culture in the case of Human Hepatocarcinoma (HepG2) cells. Principal Findings Our findings establish that the intrinsic properties of ParC-coated PDMS (ParC/PDMS) influence and modulate initial extracellular matrix (ECM; here, type-I collagen) surface architecture, as compared to non-coated PDMS substratum. Morphological changes induced by the presence of ParC on PDMS were shown to directly affect liver cell metabolic activity and the expression of transmembrane receptors implicated in cell adhesion and cell-cell interaction. These changes were characterized by atomic force microscopy (AFM), which elucidated differences in HepG2 cell adhesion, spreading, and reorganization into two- or three-dimensional structures by neosynthesis of ECM components. Local modulation of cell aggregation was successfully performed using ParC/PDMS micropatterns constructed by simple microfabrication. Conclusion/Significance We demonstrated for the first time the modulation of HepG2 cells' behavior in relation to the intrinsic physical properties of PDMS and ParC, enabling the local modulation of cell spreading in a 2D or 3D manner by simple microfabrication techniques. This work will provide promising insights into the development of cell-based platforms that have many applications in the field of in vitro liver tissue engineering, pharmacology and therapeutics. PMID:20300511

  12. Oestradiol and progesterone differentially alter cytoskeletal protein expression and flame cell morphology in Taenia crassiceps.

    PubMed

    Ambrosio, Javier R; Ostoa-Saloma, Pedro; Palacios-Arreola, M Isabel; Ruíz-Rosado, Azucena; Sánchez-Orellana, Pedro L; Reynoso-Ducoing, Olivia; Nava-Castro, Karen E; Martínez-Velázquez, Nancy; Escobedo, Galileo; Ibarra-Coronado, Elizabeth G; Valverde-Islas, Laura; Morales-Montor, Jorge

    2014-09-01

    We examined the effects of oestradiol (E2) and progesterone (P4) on cytoskeletal protein expression in the helminth Taenia crassiceps - specifically actin, tubulin and myosin. These proteins assemble into flame cells, which constitute the parasite excretory system. Total protein extracts were obtained from E2- and P4-treated T. crassiceps cysticerci and untreated controls, and analysed by one- and two-dimensional protein electrophoresis, flow cytometry, immunofluorescence and videomicroscopy. Exposure of T. crassiceps cysticerci to E2 and P4 induced differential protein expression patterns compared with untreated controls. Changes in actin, tubulin and myosin expression were confirmed by flow cytometry of parasite cells and immunofluorescence. In addition, parasite morphology was altered in response to E2 and P4 versus controls. Flame cells were primarily affected at the level of the ciliary tuft, in association with the changes in actin, tubulin and myosin. We conclude that oestradiol and progesterone act directly on T. crassiceps cysticerci, altering actin, tubulin and myosin expression and thus affecting the assembly and function of flame cells. Our results increase our understanding of several aspects of the molecular crosstalk between host and parasite, which might be useful in designing anthelmintic drugs that exclusively impair parasitic proteins which mediate cell signaling and pathogenic reproduction and establishment.

  13. Morphological variability in tree root architecture indirectly affects coexistence among competitors in the understory.

    PubMed

    Aschehoug, Erik T; Callaway, Ragan M

    2014-07-01

    Interactions between plants can have strong effects on community structure and function. Variability in the morphological, developmental, physiological, and biochemical traits of plants can influence the outcome of plant interactions and thus have important ecological consequences. However, the ecological ramifications of trait variability in plants are poorly understood and have rarely been tested in the field. We experimentally tested the effects of morphological variation in root architecture of Quercus douglasii trees in the field on interactions between understory plants and community composition. Our results indicate that variability among Q. douglasii tree root systems initiates a striking reversal in the competitive effects of dominant understory grass species on a less common species. Trees with a deep-rooted morphology facilitated exotic annual grasses and these annual grasses, in turn, competitively excluded the native perennial bunchgrass, Stipapulchra. In contrast, Q. douglasii trees with shallow-rooted morphologies directly suppressed the growth of exotic annual grasses and indirectly released S. pulchra individuals from competition with these annual grasses. Morphological variation in the root architecture of Q. douglasii created substantial conditionality in the outcomes of competition among species which enhanced the potential for indirect interactions to sustain coexistence and increase community diversity.

  14. Phenomena affecting morphology of microporous poly(acrylonitrile) prepared via phase separation from solution

    SciTech Connect

    Legasse, R.R.; Weagley, R.J.; Leslie, P.K.; Schneider, D.A.

    1990-01-01

    This paper is concerned with controlling the morphology of microporous polymers prepared via thermal demixing of solutions. 2 wt % solutions of poly(acrylonitrile) in maleic anhydride, a poor solvent, are first cooled to produce separated polymer-rich and solvent-rich phases. Removing the solvent by freeze drying then produces a microporous material having a density of 33 mg/cm{sup 3}, a void fraction of 97%, and a pore size of about 10 {mu}m. We find that the morphology cannot be explained by existing models, which focus on phase diagrams and kinetics of phase transformations during cooling of the solution. In conflict with those models, we find that two radically different morphologies can be produced even when the polymer concentration and cooling path are held strictly constant. A hypothesis that polymer degradation causes the different morphologies is not supported by GPC, {sup 13}C NMR, and FTIR experiments. Instead, we offer evidence that the different microporous morphologies are caused by different polymer conformations in solutions having the same concentration and temperature. 11 refs., 3 figs.

  15. Common polymorphisms in WNT10A affect tooth morphology as well as hair shape.

    PubMed

    Kimura, Ryosuke; Watanabe, Chiaki; Kawaguchi, Akira; Kim, Yong-Il; Park, Soo-Byung; Maki, Koutaro; Ishida, Hajime; Yamaguchi, Tetsutaro

    2015-05-01

    Hair and teeth are appendages of ectodermal origin, and there are common molecular backgrounds involved in their formation. To date, it has been revealed that a non-synonymous polymorphism in EDAR has effects on the morphological variation in both hair and teeth. Previous association studies have confirmed that single-nucleotide polymorphisms (SNPs) in/near THADA, FRAS1, WNT10A, NAF1 and FGFR2 are associated with hair morphology. In this study, we thus examined whether these SNPs are also associated with dental characteristics. We measured metric dental traits including crown size and also evaluated non-metric dental traits using plaster casts obtained from subjects (272 Japanese and 226 Koreans). DNA samples were prepared from the subjects and genotyped for the hair morphology-associated SNPs. We observed a significant association of crown size with an SNP in WNT10A (rs7349332), but not with SNPs in other genes. Therefore, we further examined four SNPs within and around WNT10A, among which rs10177996 had the strongest association with dental traits. World distribution of the derived allele in rs10177996, which is associated with larger teeth, showed that Eurasians have a higher allele frequency than Africans. Together with previous studies on hair morphology, this study demonstrated that common variations in WNT10A have pleiotropic effects on the morphology of ectodermal appendages.

  16. Effect of microfabricated microgroove-surface devices on the morphology of mesenchymal stem cells.

    PubMed

    Zhang, Xiangkai; Aoyama, Tomoki; Yasuda, Takashi; Oike, Makoto; Ito, Akira; Tajino, Junichi; Nagai, Momoko; Fujioka, Rune; Iijima, Hirotaka; Yamaguchi, Shoki; Kakinuma, Norihiro; Kuroki, Hiroshi

    2015-12-01

    The surface of a material that is in contact with cells is known to affect cell morphology and function. To develop an appropriate surface for tendon engineering, we used zigzag microgroove surfaces, which are similar to the tenocyte microenvironment. The purpose of this study was to investigate the effect of microgroove surfaces with different ridge angles (RAs), ridge lengths (RLs), ridge widths (RWs), and groove widths (GWs) on human bone marrow-derived mesenchymal stem cell (MSC) shape. Dishes with microgroove surfaces were fabricated using cyclic olefin polymer by injection-compression molding. The other parameters were fixed, and effects of different RAs (180 - 30 °), RLs (5 - 500 μm), RWs (5 - 500 μm), and GWs (5 - 500 μm) were examined. Changes in the zigzag shape of the cell due to different RAs, RLs, RWs, and GWs were observed by optical microscopy and scanning electron microscopy. Cytoskeletal changes were investigated using Phalloidin immunofluorescence staining. As observed by optical microscopy, MSCs changed to a zigzag shape in response to microgroove surfaces with different ridge and groove properties. . As observed by scanning electron microscopy, the cell shape changed at turns in the microgroove surface. Phalloidin immunofluorescence staining indicated that F-actin, not only in cell filopodia but also inside the cell body, changed orientation to conform to the microgrooves. In conclusion, the use of zigzag microgroove surfaces microfabricated by injection-compression molding demonstrated the property of MSCs to alter their shapes to fit the surface.

  17. Effect of microfabricated microgroove-surface devices on the morphology of mesenchymal stem cells.

    PubMed

    Zhang, Xiangkai; Aoyama, Tomoki; Yasuda, Takashi; Oike, Makoto; Ito, Akira; Tajino, Junichi; Nagai, Momoko; Fujioka, Rune; Iijima, Hirotaka; Yamaguchi, Shoki; Kakinuma, Norihiro; Kuroki, Hiroshi

    2015-12-01

    The surface of a material that is in contact with cells is known to affect cell morphology and function. To develop an appropriate surface for tendon engineering, we used zigzag microgroove surfaces, which are similar to the tenocyte microenvironment. The purpose of this study was to investigate the effect of microgroove surfaces with different ridge angles (RAs), ridge lengths (RLs), ridge widths (RWs), and groove widths (GWs) on human bone marrow-derived mesenchymal stem cell (MSC) shape. Dishes with microgroove surfaces were fabricated using cyclic olefin polymer by injection-compression molding. The other parameters were fixed, and effects of different RAs (180 - 30 °), RLs (5 - 500 μm), RWs (5 - 500 μm), and GWs (5 - 500 μm) were examined. Changes in the zigzag shape of the cell due to different RAs, RLs, RWs, and GWs were observed by optical microscopy and scanning electron microscopy. Cytoskeletal changes were investigated using Phalloidin immunofluorescence staining. As observed by optical microscopy, MSCs changed to a zigzag shape in response to microgroove surfaces with different ridge and groove properties. . As observed by scanning electron microscopy, the cell shape changed at turns in the microgroove surface. Phalloidin immunofluorescence staining indicated that F-actin, not only in cell filopodia but also inside the cell body, changed orientation to conform to the microgrooves. In conclusion, the use of zigzag microgroove surfaces microfabricated by injection-compression molding demonstrated the property of MSCs to alter their shapes to fit the surface. PMID:26573821

  18. Live-cell imaging study of mitochondrial morphology in mammalian cells exposed to X-rays.

    PubMed

    Noguchi, M; Kanari, Y; Yokoya, A; Narita, A; Fujii, K

    2015-09-01

    Morphological changes in mitochondria induced by X-irradiation in normal murine mammary gland cells were studied with a live-cell microscopic imaging technique. Mitochondria were visualised by staining with a specific fluorescent probe in the cells, which express fluorescent ubiquitination-based cell-cycle indicator 2 (Fucci2) probes to visualise cell cycle. In unirradiated cells, the number of cells with fragmented mitochondria was about 20 % of the total cells through observation period (96 h). In irradiated cells, the population with fragmented mitochondria significantly increased depending on the absorbed dose. Particularly, for 8 Gy irradiation, the accumulation of fragmentation persists even in the cells whose cell cycle came to a stand (80 % in G1 (G0-like) phase). The fraction reached to a maximum at 96 h after irradiation. The kinetics of the fraction with fragmented mitochondria was similar to that for cells in S/G2/M phase (20 %) through the observation period (120 h). The evidences show that, in irradiated cells, some signals are continually released from a nucleus or cytoplasm even in the G0-like cells to operate some sort of protein machineries involved in mitochondrial fission. It is inferred that this delayed mitochondrial fragmentation is strongly related to their dysfunction, and hence might modulate radiobiological effects such as mutation or cell death.

  19. Human cells and cell membrane molecular models are affected in vitro by chlorpromazine.

    PubMed

    Suwalsky, Mario; Villena, Fernando; Sotomayor, Carlos P; Bolognin, Silvia; Zatta, Paolo

    2008-06-01

    This study presents evidence that chlorpromazine (CPZ) affects human cells and cell membrane molecular models. Human SH-SY5Y neuroblastoma cells incubated with 0.1 mM CPZ suffered a decrease of cell viability. On the other hand, phase contrast microscopy observations of human erythrocytes indicated that they underwent a morphological alteration as 1 microM CPZ changed their discoid normal shape to stomatocytes, and to hemolysis with 1 mM CPZ. X-ray diffraction experiments performed on dimyristoylphosphatidylcholine (DMPC) and dimyristoylphosphatidylethanolamine (DMPE) bilayers, classes of the major phospholipids present in the outer and inner sides of the erythrocyte membrane, respectively showed that CPZ disordered the polar head and acyl chain regions of both DMPC and DMPE, where these interactions were stronger with DMPC bilayers. Fluorescence spectroscopy on DMPC LUV at 18 degrees C confirmed these results. In fact, the assays showed that CPZ induced a significant reduction of their generalized polarization (GP) and anisotropy (r) values, indicative of enhanced disorder at the polar head and acyl chain regions of the DMPC lipid bilayer. PMID:18372093

  20. Morphological characterization of keratoconus-affected human corneas by SHG imaging and correlation analysis

    NASA Astrophysics Data System (ADS)

    Mercatelli, R.; Ratto, F.; Tatini, F.; Rossi, F.; Menabuoni, L.; Nicoletti, R.; Pini, R.; Pavone, Frederick; Cicchi, R.

    2016-03-01

    Keratoconus is an ophthalmic disease in which the cornea acquires an abnormal conical shape that prevents the correct focusing on the retina, causing visual impairment. The late diagnosis of keratoconus is among the principal causes of corneal transplantation surgery. In this study, we characterize the morphology of keratoconic corneas by means of the correlation of SHG images, finding that keratoconus can be diagnosed by analyzing the inclination of lamellae below Bowman's membrane. In addition, imaging performed with both sagittal and "en face" geometry demonstrated that this morphological features can be highlighted both ex vivo and in vivo.

  1. Morphological changes in a human scirrhous gastric carcinoma cell line (KATO-III) when cultured in collagen-coated dishes.

    PubMed

    Yamamoto, R; Tatsuta, M; Nakamura, H; Matsusaka, T; Terada, N; Tamura, H

    1988-01-01

    The morphological differences between cells of a human scirrhous gastric carcinoma cell line (KATO-III) cultured in plastic dishes and in collagen-coated dishes were examined by phase-contrast and electron microscopy. When KATO-III cells were inoculated into plastic dishes, a few cells became attached to the surface of the dishes and the rest remained in suspension. However, when they were inoculated into collagen-coated dishes, they all remained in suspension. In both types of dish, most of the cells in suspension were single although a few were in clusters. The cells in suspension in collagen-coated dishes differed in morphology from those in the plastic dishes. They had abundant cytoplasm, well-developed Golgi complexes, and many microvillus-like cell protrusions. Moreover, they had hemidesmosome-like and desmosome-like structures on their surface and an increased amount of intracytoplasmic desmosome-like structures. The cells in clusters in the collagen-coated dishes were closely connected by junctional complexes, such as tight junctions, desmosomes and interdigitations, whereas those in plastic dishes were linked only by desmosomes. These results suggest that collagen affects the morphology of human scirrhous carcinoma cells. PMID:2900577

  2. Effects of adult dysthyroidism on the morphology of hippocampal granular cells in rats.

    PubMed

    Martí-Carbonell, Maria Assumpció; Garau, Adriana; Sala-Roca, Josefina; Balada, Ferran

    2012-01-01

    Thyroid hormones are essential for normal brain development and very important in the normal functioning of the brain. Thyroid hormones action in the adult brain has not been widely studied. The effects of adult hyperthyroidism are not as well understood as adult hypothyroidism, mainly in hippocampal granular cells. The purpose of the present study is to assess the consequences of adult hormone dysthyroidism (excess/deficiency of TH) on the morphology of dentate granule cells in the hippocampus by performing a quantitative study of dendritic arborizations and dendritic spines using Golgi impregnated material. Hypo-and hyperthyroidism were induced in rats by adding 0.02 percent methimazole and 1 percent L-thyroxine, respectively, to drinking water from 40 days of age. At 89 days, the animals' brains were removed and stained by a modified Golgi method and blood samples were collected in order to measure T4 serum levels. Neurons were selected and drawn using a camera lucida. Our results show that both methimazole and thyroxine treatment affect granule cell morphology. Treatments provoke alterations in the same direction, namely, reduction of certain dendritic-branching parameters that are more evident in the methimazole than in the thyroxine group. We also observe a decrease in spine density in both the methimazole and thyroxine groups. PMID:23093010

  3. Morphological Spectrum of Basal Cell Carcinoma in Southern Karnataka

    PubMed Central

    Lobo, Flora Dorothy; Naik, Ramdas; Khadilkar, Urmila Niranjan; Kini, Hema; Kini, Ullal Anand

    2016-01-01

    Introduction Basal Cell Carcinoma (BCC) is the most common skin cancer worldwide, which appears over sun-exposed skin as slow-growing, locally invasive lesion that rarely metastasizes. Many phenotypic presentations are possible. BCCs are more common in males and tend to occur in older people. Majority is found on the head and neck. Many histopathological subtypes have been defined including nodular, micronodular, cystic, superficial, pigmented, adenoid, infiltrating, sclerosing, keratotic, infundibulocystic, metatypical, basosquamous and fibroepitheliomatous. Mixed patterns are common. Aim The aim was to study morphological spectrum of BCC in a tertiary care hospital in southern Karnataka. Materials and Methods This was a retrospective analysis of 100 cases of BCCs reported in the Department of Pathology over a 9-year period from 2006 to 2014. Results The mean age of presentation was 62 years. There was slight female preponderance (56%). The most common location was face (65%) and the most common presentation was ulceration (45%). Of the 100 BCCs, 50% were nodular, 13% infiltrating, 6% basosquamous, 4% superficial, 3% keratotic, 3% multinodular and 1% mixed. Conclusion BCC, besides being the commonest cutaneous cancer, is also known for its numerous histological patterns which are shown to have prognostic implications. This study reveals the frequency of the various histological patterns of BCC in southern Karnataka, where it has been rarely studied before. PMID:27504291

  4. Glufosinate does not affect floral morphology and pollen viability in glufosinate-resistant cotton (Gossypium hirsutum)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Studies were conducted to determine whether glufosinate treatments to glufosinate-resistant cotton caused changes in floral morphology, pollen viability, and seed set. Four glufosinate treatments were included: (1) glufosinate applied postemergence over the top (POST) at the four-leaf stage, (2) glu...

  5. Androgens Exert a Cysticidal Effect upon Taenia crassiceps by Disrupting Flame Cell Morphology and Function.

    PubMed

    Ambrosio, Javier R; Valverde-Islas, Laura; Nava-Castro, Karen E; Palacios-Arreola, M Isabel; Ostoa-Saloma, Pedro; Reynoso-Ducoing, Olivia; Escobedo, Galileo; Ruíz-Rosado, Azucena; Dominguez-Ramírez, Lenin; Morales-Montor, Jorge

    2015-01-01

    The effects of testosterone (T4) and dihydrotestosterone (DHT) on the survival of the helminth cestode parasite Taenia crassiceps, as well as their effects on actin, tubulin and myosin expression and their assembly into the excretory system of flame cells are described in this paper. In vitro evaluations on parasite viability, flow cytometry, confocal microscopy, video-microscopy of live flame cells, and docking experiments of androgens interacting with actin, tubulin, and myosin were conducted. Our results show that T4 and DHT reduce T. crassiceps viability in a dose- and time-dependent fashion, reaching 90% of mortality at the highest dose used (40 ng/ml) and time exposed (10 days) in culture. Androgen treatment does not induce differences in the specific expression pattern of actin, tubulin, and myosin isoforms as compared with control parasites. Confocal microscopy demonstrated a strong disruption of the parasite tegument, with reduced assembly, shape, and motion of flame cells. Docking experiments show that androgens are capable of affecting parasite survival and flame cell morphology by directly interacting with actin, tubulin and myosin without altering their protein expression pattern. We show that both T4 and DHT are able to bind actin, tubulin, and myosin affecting their assembly and causing parasite intoxication due to impairment of flame cell function. Live flame cell video microscopy showing a reduced motion as well changes in the shape of flame cells are also shown. In summary, T4 and DHT directly act on T. crassiceps cysticerci through altering parasite survival as well as the assembly and function of flame cells.

  6. Androgens Exert a Cysticidal Effect upon Taenia crassiceps by Disrupting Flame Cell Morphology and Function

    PubMed Central

    Ambrosio, Javier R.; Valverde-Islas, Laura; Nava-Castro, Karen E.; Palacios- Arreola, M. Isabel; Ostoa-Saloma, Pedro; Reynoso-Ducoing, Olivia; Escobedo, Galileo; Ruíz-Rosado, Azucena; Dominguez-Ramírez, Lenin; Morales-Montor, Jorge

    2015-01-01

    The effects of testosterone (T4) and dihydrotestosterone (DHT) on the survival of the helminth cestode parasite Taenia crassiceps, as well as their effects on actin, tubulin and myosin expression and their assembly into the excretory system of flame cells are described in this paper. In vitro evaluations on parasite viability, flow cytometry, confocal microscopy, video-microscopy of live flame cells, and docking experiments of androgens interacting with actin, tubulin, and myosin were conducted. Our results show that T4 and DHT reduce T. crassiceps viability in a dose- and time-dependent fashion, reaching 90% of mortality at the highest dose used (40 ng/ml) and time exposed (10 days) in culture. Androgen treatment does not induce differences in the specific expression pattern of actin, tubulin, and myosin isoforms as compared with control parasites. Confocal microscopy demonstrated a strong disruption of the parasite tegument, with reduced assembly, shape, and motion of flame cells. Docking experiments show that androgens are capable of affecting parasite survival and flame cell morphology by directly interacting with actin, tubulin and myosin without altering their protein expression pattern. We show that both T4 and DHT are able to bind actin, tubulin, and myosin affecting their assembly and causing parasite intoxication due to impairment of flame cell function. Live flame cell video microscopy showing a reduced motion as well changes in the shape of flame cells are also shown. In summary, T4 and DHT directly act on T. crassiceps cysticerci through altering parasite survival as well as the assembly and function of flame cells. PMID:26076446

  7. Glial cell morphological and density changes through the lifespan of rhesus macaques.

    PubMed

    Robillard, Katelyn N; Lee, Kim M; Chiu, Kevin B; MacLean, Andrew G

    2016-07-01

    How aging impacts the central nervous system (CNS) is an area of intense interest. Glial morphology is known to affect neuronal and immune function as well as metabolic and homeostatic balance. Activation of glia, both astrocytes and microglia, occurs at several stages during development and aging. The present study analyzed changes in glial morphology and density through the entire lifespan of rhesus macaques, which are physiologically and anatomically similar to humans. We observed apparent increases in gray matter astrocytic process length and process complexity as rhesus macaques matured from juveniles through adulthood. These changes were not attributed to cell enlargement because they were not accompanied by proportional changes in soma or process volume. There was a decrease in white matter microglial process length as rhesus macaques aged. Aging was shown to have a significant effect on gray matter microglial density, with a significant increase in aged macaques compared with adults. Overall, we observed significant changes in glial morphology as macaques age indicative of astrocytic activation with subsequent increase in microglial density in aged macaques. PMID:26851132

  8. Mesenchymal stem cell adhesion but not plasticity is affected by high substrate stiffness

    NASA Astrophysics Data System (ADS)

    Kal Van Tam, Janice; Uto, Koichiro; Ebara, Mitsuhiro; Pagliari, Stefania; Forte, Giancarlo; Aoyagi, Takao

    2012-12-01

    The acknowledged ability of synthetic materials to induce cell-specific responses regardless of biological supplies provides tissue engineers with the opportunity to find the appropriate materials and conditions to prepare tissue-targeted scaffolds. Stem and mature cells have been shown to acquire distinct morphologies in vitro and to modify their phenotype when grown on synthetic materials with tunable mechanical properties. The stiffness of the substrate used for cell culture is likely to provide cells with mechanical cues mimicking given physiological or pathological conditions, thus affecting the biological properties of cells. The sensitivity of cells to substrate composition and mechanical properties resides in multiprotein complexes called focal adhesions, whose dynamic modification leads to cytoskeleton remodeling and changes in gene expression. In this study, the remodeling of focal adhesions in human mesenchymal stem cells in response to substrate stiffness was followed in the first phases of cell-matrix interaction, using poly-ɛ-caprolactone planar films with similar chemical composition and different elasticity. As compared to mature dermal fibroblasts, mesenchymal stem cells showed a specific response to substrate stiffness, in terms of adhesion, as a result of differential focal adhesion assembly, while their multipotency as a bulk was not significantly affected by matrix compliance. Given the sensitivity of stem cells to matrix mechanics, the mechanobiology of such cells requires further investigations before preparing tissue-specific scaffolds.

  9. Morphology and Performance of Polymer Solar Cell Characterized by DPD Simulation and Graph Theory.

    PubMed

    Du, Chunmiao; Ji, Yujin; Xue, Junwei; Hou, Tingjun; Tang, Jianxin; Lee, Shuit-Tong; Li, Youyong

    2015-01-01

    The morphology of active layers in the bulk heterojunction (BHJ) solar cells is critical to the performance of organic photovoltaics (OPV). Currently, there is limited information for the morphology from transmission electron microscopy (TEM) techniques. Meanwhile, there are limited approaches to predict the morphology /efficiency of OPV. Here we use Dissipative Particle Dynamics (DPD) to determine 3D morphology of BHJ solar cells and show DPD to be an efficient approach to predict the 3D morphology. Based on the 3D morphology, we estimate the performance indicator of BHJ solar cells by using graph theory. Specifically, we study poly (3-hexylthiophene)/[6, 6]-phenyl-C61butyric acid methyl ester (P3HT/PCBM) BHJ solar cells. We find that, when the volume fraction of PCBM is in the region 0.4 ∼ 0.5, P3HT/PCBM will show bi-continuous morphology and optimum performance, consistent with experimental results. Further, the optimum temperature (413 K) for the morphology and performance of P3HT/PCBM is in accord with annealing results. We find that solvent additive plays a critical role in the desolvation process of P3HT/PCBM BHJ solar cell. Our approach provides a direct method to predict dynamic 3D morphology and performance indicator for BHJ solar cells. PMID:26581407

  10. Morphology and Performance of Polymer Solar Cell Characterized by DPD Simulation and Graph Theory.

    PubMed

    Du, Chunmiao; Ji, Yujin; Xue, Junwei; Hou, Tingjun; Tang, Jianxin; Lee, Shuit-Tong; Li, Youyong

    2015-11-19

    The morphology of active layers in the bulk heterojunction (BHJ) solar cells is critical to the performance of organic photovoltaics (OPV). Currently, there is limited information for the morphology from transmission electron microscopy (TEM) techniques. Meanwhile, there are limited approaches to predict the morphology /efficiency of OPV. Here we use Dissipative Particle Dynamics (DPD) to determine 3D morphology of BHJ solar cells and show DPD to be an efficient approach to predict the 3D morphology. Based on the 3D morphology, we estimate the performance indicator of BHJ solar cells by using graph theory. Specifically, we study poly (3-hexylthiophene)/[6, 6]-phenyl-C61butyric acid methyl ester (P3HT/PCBM) BHJ solar cells. We find that, when the volume fraction of PCBM is in the region 0.4 ∼ 0.5, P3HT/PCBM will show bi-continuous morphology and optimum performance, consistent with experimental results. Further, the optimum temperature (413 K) for the morphology and performance of P3HT/PCBM is in accord with annealing results. We find that solvent additive plays a critical role in the desolvation process of P3HT/PCBM BHJ solar cell. Our approach provides a direct method to predict dynamic 3D morphology and performance indicator for BHJ solar cells.

  11. Morphology and Performance of Polymer Solar Cell Characterized by DPD Simulation and Graph Theory

    PubMed Central

    Du, Chunmiao; Ji, Yujin; Xue, Junwei; Hou, Tingjun; Tang, Jianxin; Lee, Shuit-Tong; Li, Youyong

    2015-01-01

    The morphology of active layers in the bulk heterojunction (BHJ) solar cells is critical to the performance of organic photovoltaics (OPV). Currently, there is limited information for the morphology from transmission electron microscopy (TEM) techniques. Meanwhile, there are limited approaches to predict the morphology /efficiency of OPV. Here we use Dissipative Particle Dynamics (DPD) to determine 3D morphology of BHJ solar cells and show DPD to be an efficient approach to predict the 3D morphology. Based on the 3D morphology, we estimate the performance indicator of BHJ solar cells by using graph theory. Specifically, we study poly (3-hexylthiophene)/[6, 6]-phenyl-C61butyric acid methyl ester (P3HT/PCBM) BHJ solar cells. We find that, when the volume fraction of PCBM is in the region 0.4 ∼ 0.5, P3HT/PCBM will show bi-continuous morphology and optimum performance, consistent with experimental results. Further, the optimum temperature (413 K) for the morphology and performance of P3HT/PCBM is in accord with annealing results. We find that solvent additive plays a critical role in the desolvation process of P3HT/PCBM BHJ solar cell. Our approach provides a direct method to predict dynamic 3D morphology and performance indicator for BHJ solar cells. PMID:26581407

  12. Morphology and Performance of Polymer Solar Cell Characterized by DPD Simulation and Graph Theory

    NASA Astrophysics Data System (ADS)

    Du, Chunmiao; Ji, Yujin; Xue, Junwei; Hou, Tingjun; Tang, Jianxin; Lee, Shuit-Tong; Li, Youyong

    2015-11-01

    The morphology of active layers in the bulk heterojunction (BHJ) solar cells is critical to the performance of organic photovoltaics (OPV). Currently, there is limited information for the morphology from transmission electron microscopy (TEM) techniques. Meanwhile, there are limited approaches to predict the morphology /efficiency of OPV. Here we use Dissipative Particle Dynamics (DPD) to determine 3D morphology of BHJ solar cells and show DPD to be an efficient approach to predict the 3D morphology. Based on the 3D morphology, we estimate the performance indicator of BHJ solar cells by using graph theory. Specifically, we study poly (3-hexylthiophene)/[6, 6]-phenyl-C61butyric acid methyl ester (P3HT/PCBM) BHJ solar cells. We find that, when the volume fraction of PCBM is in the region 0.4 ∼ 0.5, P3HT/PCBM will show bi-continuous morphology and optimum performance, consistent with experimental results. Further, the optimum temperature (413 K) for the morphology and performance of P3HT/PCBM is in accord with annealing results. We find that solvent additive plays a critical role in the desolvation process of P3HT/PCBM BHJ solar cell. Our approach provides a direct method to predict dynamic 3D morphology and performance indicator for BHJ solar cells.

  13. Egestion of asbestos fibers in Tetrahymena results in early morphological abnormalities. A step in the induction of heterogeneous cell lines?

    PubMed

    Hjelm, K K

    1989-01-01

    In Tetrahymena populations exposed to crocidolite asbestos fibers, many cells develop morphological abnormalities within 1-2 hours. The abnormalities are mainly large or small protrusions or indentations, or flattened parts of the cell surface and most often located in the posterior part of the cell. They are formed repeatedly in all cells but are also continuously repaired so that the fraction of cells affected represents an equilibrium between these two processes. Their formation is connected with egestion of the large bundles of fibers formed by phagocytosis. Such effects of egestion of fibers do not seem to have been reported previously. Egestion of a bundle of fibers is much slower than for other types of undigestible residues. In contrast to normal exocytosis occurring invariably at the cytoproct, egestion of asbestos often occurs in the posterior part of the cell outside the cytoproct. To my knowledge this is the first reported case of either very slow or extra-cytoproctal egestion in Tetrahymena. Cells with large abnormalities have a greater tendency to develop into "early heterogeneous" cells than the average abnormal cell. Some of these give rise to hereditarily stable heterogeneous cell lines of Tetrahymena. The morphological abnormalities are probably caused by mechanical action of the crocidolite fibers resulting in local damage of the cytoskeletal elements responsible for normal cell shape. The heterogenous cell lines may arise when cellular structures carrying non-genic cytotactically inherited information are modified. The relevance of these ideas to the induction of cancer by asbestos is briefly discussed.

  14. How Will Climate Change Affect Channel Morphology and Salmonid Habitat in Mountain Basins?

    NASA Astrophysics Data System (ADS)

    Buffington, J. M.; Goode, J.

    2010-12-01

    Riverine habitat for salmonids is intimately linked to channel morphology and fluvial processes (channel hydraulics, sediment transport and scour regime) which are, in turn, controlled by watershed hydrology and erosional processes that input sediment to the fluvial system. Climate change has the potential to alter the timing, magnitude, and style of sediment and water inputs to mountain rivers. Channel response to these changes may range from small-scale adjustments of channel characteristics (e.g., width, depth, grain size, scour depth) to larger-scale changes in channel type (e.g., metamorphosis from a pool-riffle channel to a plane-bed morphology). Identifying which parts of the river network will remain relatively stable in response to climate change, and which are likely to cross critical morphologic and scour thresholds is important for predicting effects on salmonid populations. Toward this end, a regime framework is presented for predicting the relative degree of morphologic stability and scour potential in different physiographic settings (different water and sediment regimes). Digital elevation models are used to explore the spatial distribution of these conditions and potential consequences for salmonid habitat across the landscape. Results suggest that the potential for scour and morphologic variability are strongly influenced by hydroclimate; snowmelt channels are relatively stable across floods of different magnitude, while rainfall-dominated channels are more variable and less stable. Transitional changes in hydrologic regime (mixed rain and snow) have the greatest potential for altering geomorphic conditions and salmonid habitat. However, the vulnerability of salmonids to climate-driven changes in scour regime depend on the species and its life history (i.e., depth to which eggs are buried and timing of incubation relative to scouring flows). Overall, the regime approach provides a useful first-order assessment of channel condition and response

  15. Cartography of cell morphology in tomato pericarp at the fruit scale.

    PubMed

    Legland, D; Devaux, M-F; Bouchet, B; Guillon, F; Lahaye, M

    2012-07-01

    In fleshy fruits, the variability of cell morphology at the fruit scale is largely unknown. It presents both a huge variability and a high level of organization. Better knowledge of cell morphology heterogeneity within the fruit is necessary to understand fruit development, to model fruit mechanical behaviour, or to investigate variations of physico-chemical measurements. A generic approach is proposed to build cartographies of cell morphology at the fruit scale, which depict regions corresponding to different cell morphologies. The approach is based on: (1) sampling the whole fruit at known positions; (2) imaging and quantifying local cell morphology; (3) pooling measurements to take biological variability into account and (4) projecting results in a morphology model of the whole fruit. The result is a synthetic representation of cell morphology variations within the whole fruit. The method was applied to the characterization of cell morphology in tomato pericarp. Two different imaging scales that provided complementary descriptions were used: 3D confocal microscopy and macroscopy. The approach is generic and can be adapted to other fruits or other products.

  16. Metformin selectively affects human glioblastoma tumor-initiating cell viability

    PubMed Central

    Würth, Roberto; Pattarozzi, Alessandra; Gatti, Monica; Bajetto, Adirana; Corsaro, Alessandro; Parodi, Alessia; Sirito, Rodolfo; Massollo, Michela; Marini, Cecilia; Zona, Gianluigi; Fenoglio, Daniela; Sambuceti, Gianmario; Filaci, Gilberto; Daga, Antonio; Barbieri, Federica; Florio, Tullio

    2013-01-01

    Cancer stem cell theory postulates that a small population of tumor-initiating cells is responsible for the development, progression and recurrence of several malignancies, including glioblastoma. In this perspective, tumor-initiating cells represent the most relevant target to obtain effective cancer treatment. Metformin, a first-line drug for type II diabetes, was reported to possess anticancer properties affecting the survival of cancer stem cells in breast cancer models. We report that metformin treatment reduced the proliferation rate of tumor-initiating cell-enriched cultures isolated from four human glioblastomas. Metformin also impairs tumor-initiating cell spherogenesis, indicating a direct effect on self-renewal mechanisms. Interestingly, analyzing by FACS the antiproliferative effects of metformin on CD133-expressing subpopulation, a component of glioblastoma cancer stem cells, a higher reduction of proliferation was observed as compared with CD133-negative cells, suggesting a certain degree of cancer stem cell selectivity in its effects. In fact, glioblastoma cell differentiation strongly reduced sensitivity to metformin treatment. Metformin effects in tumor-initiating cell-enriched cultures were associated with a powerful inhibition of Akt-dependent cell survival pathway, while this pathway was not affected in differentiated cells. The specificity of metformin antiproliferative effects toward glioblastoma tumor-initiating cells was confirmed by the lack of significant inhibition of normal human stem cells (umbilical cord-derived mesenchymal stem cells) in vitro proliferation after metformin exposure. Altogether, these data clearly suggest that metformin exerts antiproliferative activity on glioblastoma cells, showing a higher specificity toward tumor-initiating cells, and that the inhibition of Akt pathway may represent a possible intracellular target of this effect. PMID:23255107

  17. Nanoscale Roughness and Morphology Affect the IsoElectric Point of Titania Surfaces

    PubMed Central

    Borghi, Francesca; Vyas, Varun; Podestà, Alessandro; Milani, Paolo

    2013-01-01

    We report on the systematic investigation of the role of surface nanoscale roughness and morphology on the charging behaviour of nanostructured titania (TiO2) surfaces in aqueous solutions. IsoElectric Points (IEPs) of surfaces have been characterized by direct measurement of the electrostatic double layer interactions between titania surfaces and the micrometer-sized spherical silica probe of an atomic force microscope in NaCl aqueous electrolyte. The use of a colloidal probe provides well-defined interaction geometry and allows effectively probing the overall effect of nanoscale morphology. By using supersonic cluster beam deposition to fabricate nanostructured titania films, we achieved a quantitative control over the surface morphological parameters. We performed a systematical exploration of the electrical double layer properties in different interaction regimes characterized by different ratios of characteristic nanometric lengths of the system: the surface rms roughness Rq, the correlation length ξ and the Debye length λD. We observed a remarkable reduction by several pH units of IEP on rough nanostructured surfaces, with respect to flat crystalline rutile TiO2. In order to explain the observed behavior of IEP, we consider the roughness-induced self-overlap of the electrical double layers as a potential source of deviation from the trend expected for flat surfaces. PMID:23874708

  18. AGING AFFECTS MORPHOLOGY BUT NOT STIMULATED SECRETION OF SALIVA IN RATS

    PubMed Central

    Lasisi, T.J.; Shittu, S.T.; Oguntokun, M.M.; Tiamiyu, N.A.

    2014-01-01

    Background: The role of aging on the salivary gland function still remains controversial and inconclusive. This study was undertaken to determine the effects of aging on the morphology and secretion of salivary glands using male Wistar rats. Method: There were three age groups; group A (3 months old; n = 8), group B (6 months old; n = 8), and group C (9 months old; n = 8). Body weights, salivary gland weights, salivary flow rates, pH and salivary levels of sodium, potassium, calcium, chloride, bicarbonate, phosphate and total protein were measured and compared. Hematoxylin-eosin stained histological slides of the salivary glands were assessed for morphological changes. Results: Body weights increased with age while mean parotid gland weight was significantly higher in group B than in groups A and C. Mean salivary flow rate was significantly higher in group B and C than in group A, and mean salivary pH was significantly higher in group B and C than group A. Analysis of salivary electrolytes and total protein showed that mean levels of sodium, potassium and bicarbonate increased with age significantly while mean levels of calcium, chloride, phosphate and total protein did not show significant change among the groups. Conclusion: These findings showed that varying changes were observed in the morphology of salivary glands of aging rats without impaired function. PMID:25960701

  19. A quantitative study of MC3T3-E1 cell adhesion, morphology and biomechanics on chitosan-collagen blend films at single cell level.

    PubMed

    Wang, Chuang; Xie, Xu-dong; Huang, Xun; Liang, Zhi-hong; Zhou, Chang-ren

    2015-08-01

    The interaction between cells and biomaterials plays a key role in cell proliferation and differentiation in tissue engineering. However, a quantitative analysis of those interactions has been less well studied. The objective of this study was to quantitative recapitulate the difference of MC3T3-E1 cell adhesion, morphological and biomechanical properties on chitosan-collagen films in terms of chemical composition. Here, the unbinding force between MC3T3-E1 cell and a series of chitosan-collagen films was probed by a real-time and in situ atomic force microscopy-single cell force spectroscopy (AFM-SCFS). Meanwhile, changes in cell morphology and Young's modulus on different chitosan-collagen films were detected by AFM. The cell area and CCK-8 results showed that cell spreading and proliferation increased with increasing collagen content. AFM observations clearly showed cell height decreased and pseudopod fusion with the collagen content increased. Cell adhesive force increased from 0.76±0.17 nN to 1.70±0.19 nN. On the contrary, cells Young's modulus, which reflected biophysical changes of cells decreased from 11.94±3.19 kPa to 1.81±0.52 kPa, respectively. It suggested that stronger cell-substrate interactions benefit cell adhesion, and better cell flexibility improve cell spreading. The findings indicate that cell morphology, adhesive force and Young's modulus are significant affected by various chitosan-collagen substrates. Those methods and quantitative results have guiding significance for investigating the mechanism of chitosan and/or collagen based cell-targeting drug carrier and the preparation of chitosan-collagen composite biomaterials.

  20. Changes in Cell Morphology Are Coordinated with Cell Growth through the TORC1 Pathway

    PubMed Central

    Goranov, Alexi I.; Gulati, Amneet; Dephoure, Noah; Takahara, Terunao; Maeda, Tatsuya; Gygi, Steven P.; Manalis, Scott; Amon, Angelika

    2013-01-01

    Summary Background Growth rate is determined not only by extracellular cues such as nutrient availability but also by intracellular processes. Changes in cell morphology in budding yeast, mediated by polarization of the actin cytoskeleton, have been shown to reduce cell growth. Results Here we demonstrate that polarization of the actin cytoskeleton inhibits the highly conserved Target of Rapamycin Complex 1 (TORC1) pathway. This downregulation is suppressed by inactivation of the TORC1 pathway regulatory Iml1 complex, which also regulates TORC1 during nitrogen starvation. We further demonstrate that attenuation of growth is important for cell recovery after conditions of prolonged polarized growth. Conclusions Our results indicate that extended periods of polarized growth inhibit protein synthesis, mass accumulation, and the increase in cell size at least in part through inhibiting the TORC1 pathway. We speculate that this mechanism serves to coordinate the ability of cells to increase in size with their biosynthetic capacity. PMID:23810534

  1. Pinocchio cells: morphologically atypical immunologically heterogeneous lymphocytes induced by treatment with interleukin 2.

    PubMed

    Paciucci, P A; Chesa, P G; Fierro, M T; Gordon, R; Konefal, R G; Glidewell, O; Holland, J F

    1988-12-01

    We describe a novel cell type, the Pinocchio cell, that appears in the peripheral blood of all patients receiving treatment with interleukin 2, up to 20,000 cells/microliter. This cell is characterized by a prominent and granular proboscis with which it attaches to tumor cells and mediates tumor cell lysis. Pinocchio cells are immunologically heterogeneous and express antigens of both T and NK cells; Pinocchio cells are adherent in culture and are more cytolytic than non-adherent cells against NK-sensitive and resistant tumor cells. Incubation of normal whole human blood for 1 h induces Pinocchio morphology of mononuclear white blood cells.

  2. Splitting of IVP bovine blastocyst affects morphology and gene expression of resulting demi-embryos during in vitro culture and in vivo elongation.

    PubMed

    Velasquez, Alejandra E; Castro, Fidel O; Veraguas, Daniel; Cox, Jose F; Lara, Evelyn; Briones, Mario; Rodriguez-Alvarez, Lleretny

    2016-02-01

    Embryo splitting might be used to increase offspring yield and for molecular analysis of embryo competence. How splitting affects developmental potential of embryos is unknown. This research aimed to study the effect of bovine blastocyst splitting on morphological and gene expression homogeneity of demi-embryos and on embryo competence during elongation. Grade I bovine blastocyst produced in vitro were split into halves and distributed in nine groups (3 × 3 setting according to age and stage before splitting; age: days 7-9; stage: early, expanded and hatched blastocysts). Homogeneity and survival rate in vitro after splitting (12 h, days 10 and 13) and the effect of splitting on embryo development at elongation after embryo transfer (day 17) were assessed morphologically and by RT-qPCR. The genes analysed were OCT4, SOX2, NANOG, CDX2, TP1, TKDP1, EOMES, and BAX. Approximately 90% of split embryos had a well conserved defined inner cell mass (ICM), 70% of the halves had similar size with no differences in gene expression 12 h after splitting. Split embryos cultured further conserved normal and comparable morphology at day 10 of development; this situation changes at day 13 when embryo morphology and gene expression differed markedly among demi-embryos. Split and non-split blastocysts were transferred to recipient cows and were recovered at day 17. Fifty per cent of non-split embryos were larger than 100 mm (33% for split embryos). OCT4, SOX2, TP1 and EOMES levels were down-regulated in elongated embryos derived from split blastocysts. In conclusion, splitting day-8 blastocysts yields homogenous demi-embryos in terms of developmental capability and gene expression, but the initiation of the filamentous stage seems to be affected by the splitting.

  3. Potato Snakin-1 Gene Silencing Affects Cell Division, Primary Metabolism, and Cell Wall Composition1[W

    PubMed Central

    Nahirñak, Vanesa; Almasia, Natalia Inés; Fernandez, Paula Virginia; Hopp, Horacio Esteban; Estevez, José Manuel; Carrari, Fernando; Vazquez-Rovere, Cecilia

    2012-01-01

    Snakin-1 (SN1) is an antimicrobial cysteine-rich peptide isolated from potato (Solanum tuberosum) that was classified as a member of the Snakin/Gibberellic Acid Stimulated in Arabidopsis protein family. In this work, a transgenic approach was used to study the role of SN1 in planta. Even when overexpressing SN1, potato lines did not show remarkable morphological differences from the wild type; SN1 silencing resulted in reduced height, which was accompanied by an overall reduction in leaf size and severe alterations of leaf shape. Analysis of the adaxial epidermis of mature leaves revealed that silenced lines had 70% to 90% increases in mean cell size with respect to wild-type leaves. Consequently, the number of epidermal cells was significantly reduced in these lines. Confocal microscopy analysis after agroinfiltration of Nicotiana benthamiana leaves showed that SN1-green fluorescent protein fusion protein was localized in plasma membrane, and bimolecular fluorescence complementation assays revealed that SN1 self-interacted in vivo. We further focused our study on leaf metabolism by applying a combination of gas chromatography coupled to mass spectrometry, Fourier transform infrared spectroscopy, and spectrophotometric techniques. These targeted analyses allowed a detailed examination of the changes occurring in 46 intermediate compounds from primary metabolic pathways and in seven cell wall constituents. We demonstrated that SN1 silencing affects cell division, leaf primary metabolism, and cell wall composition in potato plants, suggesting that SN1 has additional roles in growth and development beyond its previously assigned role in plant defense. PMID:22080603

  4. Nephrocalcin isoforms coat crystal surfaces and differentially affect calcium oxalate monohydrate crystal morphology, growth, and aggregation

    NASA Astrophysics Data System (ADS)

    Kurutz, Josh W.; Carvalho, Mauricio; Nakagawa, Yasushi

    2003-08-01

    Calcium oxalate crystals were grown in the presence of each of the four isoforms of nephrocalcin (NC), a urinary protein proposed to inhibit kidney stone growth. Crystal size, morphology, and surface topography were assessed using optical microscopy, Coulter counter measurements, scanning electron microscopy (SEM), and atomic force microscopy (AFM). All crystals grown in the presence of NC isoforms were calcium oxalate monohydrates (COMs). Crystals formed in the presence of NC-A were smaller than control crystals, which were grown without NC, according to optical and SEM results, suggesting that NC-A restricts crystal growth. In contrast, samples grown with NC-C and NC-D exhibit more large crystals and several crystal aggregates, suggesting that NC-C and -D promote crystal growth and aggregation. Crystals grown with NC-B are not significantly larger or smaller than controls. AFM images of the crystals reveal significantly different surface textures on the control crystals relative to those grown with NC isoforms, indicating that NC acts by coating nascent calcium oxalate crystals. These are the first reported AFM images that show topography of NC-coated crystals. These findings suggest that NC isoforms have distinct interactions with different COM crystal faces, which may be responsible for their different effects on crystal growth and morphology.

  5. A pivotal role of Rho GTPase in the regulation of morphology and function of dendritic cells.

    PubMed

    Kobayashi, M; Azuma, E; Ido, M; Hirayama, M; Jiang, Q; Iwamoto, S; Kumamoto, T; Yamamoto, H; Sakurai, M; Komada, Y

    2001-10-01

    Dendritic cell (DC) is the most potent activator of CD4+ T cells and has unique dendrites and veils. To explore the function of Rho in DC, exoenzyme C3 from Clostridium botulinum was used as a specific inhibitor of Rho. Treatment of DC with C3 (DC/C3) resulted in profound morphological changes by losing dendrites and emerging of shrunk membrane processes that were in parallel with marked reduction of polymerized actin in the marginal area. Inactivation of Rho-associated coiled coil-containing kinase (p160ROCK) by a specific ROCK inhibitor Y-27632 also led to disappearance of dendrites of DC with retaining large membrane expansions. In scanning electron microscopy, untreated DCs interacted with CD4+ T cells more efficiently than DC/C3. Conjugate formation assay showed that the number of DCs associated with CD4+ T cells was 2-fold higher in untreated DCs than that of DC/C3. Alloantigen-presenting capacity of DC/C3 was significantly suppressed in a dose-dependent manner. Because C3 treatment did not affect the surface expression of HLA, costimulatory, and adhesion molecules of DC, we examined cytokine production of DC and naive CD4+ T cells to further elucidate the inhibitory mechanism of MLR. Unexpectedly, DC/C3 increased IL-12 production after LPS stimulation. Naive CD4+ T cells cocultured with DC/C3 produced the increased percentage of IFN-gamma-producing cells, whereas the percentage of IL-2-producing T cells was decreased. These results demonstrate that Rho GTPase in DC controls both characteristic shape and immunogenic capacity. PMID:11564770

  6. Morphological Criteria of Cell Differentiation Stages in Experimental Hepatocarcinoma and Evaluation of Antitumor Drug Efficiency.

    PubMed

    Bgatova, N P; Omel'yanchuk, L V; Pozhidaeva, A A; Semeshin, V F; Lykov, A P; Poveshchenko, O V; Makarova, O P; Rachkovskaya, L N; Borodin, Yu I; Konenkov, V I

    2015-11-01

    Structural polymorphism of 5 cell differentiation stages of hepatocarcinoma-29 from ascitic fluid is detected and the morphological criteria for identification of these stages are defined on the base of optic and electron microscopy findings, cytofluorometry, and DNA cytometry. The percentage of cells at differentiation stages 4 and 5 in the tumor structure increases after hepatocarcinoma cell inoculation into the hip. Injection of a cell cycle-modulating substance to animals with tumor growth shifts the proportion of cells with various differentiation stages. The morphological criteria of 5 stages of hepatocarcinoma-29 cell differentiation can be used for prospective drug testing. PMID:26593417

  7. A Study of Parameters Affecting Fibroblast Morphology in Response to an Applied Mechanical Force

    NASA Technical Reports Server (NTRS)

    Grymes, Rosalind A.; Sawyer, Christine

    1994-01-01

    A precisely controlled stretch/relaxation regimen (20% elongation at 6.6 cycles/min) was applied to normal human fetal, neonatal and aged dermal fibroblasts cultured on flexible membranes. Culture conditions included poly (NH2) or collagen type I coated substrate membranes; control cultures were grown on the same pliable material in the absence of applied stretch. Direct observation and immunofluorescence analyses revealed a progressive change in cell body orientation limited to the stretched dermal fibroblast cultures. Monolayers gradually (over 4 days) acquired a symmetric, radial distribution equivalent to the biaxial array of the applied force. At high seeding density, alignment was inhibited in the fetal cell cultures. This cell strain required collagen type I coating for optimal attachment to the flexible membrane, preferring growth in three-dimensional cell 'balls' on the poly(NH2) coated substrate. Neonatal cells also required the collagen type I coating, but both neonatal and aged dermal fibroblasts aligned efficiently at all seeding densities examined. The randomly oriented neonatal cells on the unstretched control membranes spontaneously detached at confluence, as a single cell sheet. Their aligned counterparts did not detach until the applied stretch stimulus was removed. Low concentrations of cytochalasin D (62.5 ng/ml) disrupted the stretch-related alignment response. Rhodamine phalloidin staining visualized fewer actin stress fibers in stretched, aligned cells than in controls. Both intercellular interactions and cytoskeletal integrity mediate the response to mechanical strain. Normal rabbit corneal stroma fibroblasts (NRC) were also analyzed, and failed to orient under these conditions. This cell type may require a different regimen, or a longer time period, to demonstrate alignment behavior. Supported by NASA Space Biology RTOP 199-40-22 and the NASA-ARC Director's Discretionary Fund.

  8. Sublethal concentrations of carbapenems alter cell morphology and genomic expression of Klebsiella pneumoniae biofilms.

    PubMed

    Van Laar, Tricia A; Chen, Tsute; You, Tao; Leung, Kai P

    2015-03-01

    Klebsiella pneumoniae, a Gram-negative bacterium, is normally associated with pneumonia in patients with weakened immune systems. However, it is also a prevalent nosocomial infectious agent that can be found in infected surgical sites and combat wounds. Many of these clinical strains display multidrug resistance. We have worked with a clinical strain of K. pneumoniae that was initially isolated from a wound of an injured soldier. This strain demonstrated resistance to many commonly used antibiotics but sensitivity to carbapenems. This isolate was capable of forming biofilms in vitro, contributing to its increased antibiotic resistance and impaired clearance. We were interested in determining how sublethal concentrations of carbapenem treatment specifically affect K. pneumoniae biofilms both in morphology and in genomic expression. Scanning electron microscopy showed striking morphological differences between untreated and treated biofilms, including rounding, blebbing, and dimpling of treated cells. Comparative transcriptome analysis using RNA sequencing (RNA-Seq) technology identified a large number of open reading frames (ORFs) differentially regulated in response to carbapenem treatment at 2 and 24 h. ORFs upregulated with carbapenem treatment included genes involved in resistance, as well as those coding for antiporters and autoinducers. ORFs downregulated included those coding for metal transporters, membrane biosynthesis proteins, and motility proteins. Quantitative real-time PCR validated the general trend of some of these differentially regulated ORFs. Treatment of K. pneumoniae biofilms with sublethal concentrations of carbapenems induced a wide range of phenotypic and gene expression changes. This study reveals some of the mechanisms underlying how sublethal amounts of carbapenems could affect the overall fitness and pathogenic potential of K. pneumoniae biofilm cells. PMID:25583711

  9. Sublethal Concentrations of Carbapenems Alter Cell Morphology and Genomic Expression of Klebsiella pneumoniae Biofilms

    PubMed Central

    Van Laar, Tricia A.; Chen, Tsute; You, Tao

    2015-01-01

    Klebsiella pneumoniae, a Gram-negative bacterium, is normally associated with pneumonia in patients with weakened immune systems. However, it is also a prevalent nosocomial infectious agent that can be found in infected surgical sites and combat wounds. Many of these clinical strains display multidrug resistance. We have worked with a clinical strain of K. pneumoniae that was initially isolated from a wound of an injured soldier. This strain demonstrated resistance to many commonly used antibiotics but sensitivity to carbapenems. This isolate was capable of forming biofilms in vitro, contributing to its increased antibiotic resistance and impaired clearance. We were interested in determining how sublethal concentrations of carbapenem treatment specifically affect K. pneumoniae biofilms both in morphology and in genomic expression. Scanning electron microscopy showed striking morphological differences between untreated and treated biofilms, including rounding, blebbing, and dimpling of treated cells. Comparative transcriptome analysis using RNA sequencing (RNA-Seq) technology identified a large number of open reading frames (ORFs) differentially regulated in response to carbapenem treatment at 2 and 24 h. ORFs upregulated with carbapenem treatment included genes involved in resistance, as well as those coding for antiporters and autoinducers. ORFs downregulated included those coding for metal transporters, membrane biosynthesis proteins, and motility proteins. Quantitative real-time PCR validated the general trend of some of these differentially regulated ORFs. Treatment of K. pneumoniae biofilms with sublethal concentrations of carbapenems induced a wide range of phenotypic and gene expression changes. This study reveals some of the mechanisms underlying how sublethal amounts of carbapenems could affect the overall fitness and pathogenic potential of K. pneumoniae biofilm cells. PMID:25583711

  10. Sublethal concentrations of carbapenems alter cell morphology and genomic expression of Klebsiella pneumoniae biofilms.

    PubMed

    Van Laar, Tricia A; Chen, Tsute; You, Tao; Leung, Kai P

    2015-03-01

    Klebsiella pneumoniae, a Gram-negative bacterium, is normally associated with pneumonia in patients with weakened immune systems. However, it is also a prevalent nosocomial infectious agent that can be found in infected surgical sites and combat wounds. Many of these clinical strains display multidrug resistance. We have worked with a clinical strain of K. pneumoniae that was initially isolated from a wound of an injured soldier. This strain demonstrated resistance to many commonly used antibiotics but sensitivity to carbapenems. This isolate was capable of forming biofilms in vitro, contributing to its increased antibiotic resistance and impaired clearance. We were interested in determining how sublethal concentrations of carbapenem treatment specifically affect K. pneumoniae biofilms both in morphology and in genomic expression. Scanning electron microscopy showed striking morphological differences between untreated and treated biofilms, including rounding, blebbing, and dimpling of treated cells. Comparative transcriptome analysis using RNA sequencing (RNA-Seq) technology identified a large number of open reading frames (ORFs) differentially regulated in response to carbapenem treatment at 2 and 24 h. ORFs upregulated with carbapenem treatment included genes involved in resistance, as well as those coding for antiporters and autoinducers. ORFs downregulated included those coding for metal transporters, membrane biosynthesis proteins, and motility proteins. Quantitative real-time PCR validated the general trend of some of these differentially regulated ORFs. Treatment of K. pneumoniae biofilms with sublethal concentrations of carbapenems induced a wide range of phenotypic and gene expression changes. This study reveals some of the mechanisms underlying how sublethal amounts of carbapenems could affect the overall fitness and pathogenic potential of K. pneumoniae biofilm cells.

  11. Estrogen deficiency heterogeneously affects tissue specific stem cells in mice.

    PubMed

    Kitajima, Yuriko; Doi, Hanako; Ono, Yusuke; Urata, Yoshishige; Goto, Shinji; Kitajima, Michio; Miura, Kiyonori; Li, Tao-Sheng; Masuzaki, Hideaki

    2015-01-01

    Postmenopausal disorders are frequently observed in various organs, but their relationship with estrogen deficiency and mechanisms remain unclear. As tissue-specific stem cells have been found to express estrogen receptors, we examined the hypothesis that estrogen deficiency impairs stem cells, which consequently contributes to postmenopausal disorders. Six-week-old C57BL/6 female mice were ovariectomized, following which they received 17β-estradiol replacement or vehicle (control). Sham-operated mice were used as healthy controls. All mice were killed for evaluation 2 months after treatments. Compared with the healthy control, ovariectomy significantly decreased uterine weight, which was partially recovered by 17β-estradiol replacement. Ovariectomy significantly increased the numbers of c-kit-positive hematopoietic stem/progenitor cells in bone marrow, but impaired their capacity to grow mixed cell-type colonies in vitro. Estrogen replacement further increased the numbers of c-kit-positive hematopoietic stem/progenitor cells in bone marrow, without significantly affecting colony growth in vitro. The number of CD105-positive mesenchymal stem cells in bone marrow also significantly decreased after ovariectomy, but completely recovered following estrogen replacement. Otherwise, neither ovariectomy nor estrogen replacement changed the number of Pax7-positive satellite cells, which are a skeletal muscle-type stem cell. Estrogen deficiency heterogeneously affected tissue-specific stem cells, suggesting a likely and direct relationship with postmenopausal disorders. PMID:26245252

  12. Developmental Trenbolone Exposure Affects Adult Breeding Behavior, Fecundity and Morphology of Xenopus tropicalis

    EPA Science Inventory

    Trenbolone acetate is a synthetic androgen used as a growth promoter in the cattle industry. Its metabolite 17â-trenbolone (17â-T) has been detected downstream from cattle feedlots. It could be a concern to wildlife near these areas as previous studies show 17â-T exposure affects...

  13. Bactericidal thurincin H causes unique morphological changes in Bacillus cereus F4552 without affecting membrane permeability.

    PubMed

    Wang, Gaoyan; Feng, Guoping; Snyder, Abigail B; Manns, David C; Churey, John J; Worobo, Randy W

    2014-08-01

    Thurincin H is an antilisterial bacteriocin produced by Bacillus thuringiensis SF361. It exhibits inhibitory activity against a wide range of Gram-positive foodborne pathogens and spoilage bacteria including Listeria monocytogenes, B. cereus, and B. subtilis. This hydrophobic, anionic bacteriocin folds into a hairpin structure maintained by four pairs of unique sulfur to α-carbon thioether bonds. As its hydrophobicity and structure are quite different from most archived bacteriocins, this study aimed to elucidate its mode of action and compare it with the mechanisms of other well-characterized bacteriocins. The results indicated that, although bactericidal to B. cereus F4552, thurincin H did not lead to optical density reduction or detectable changes in cell membrane permeability. B. cereus F4552 imaged by scanning electron microscopy after treatment with thurincin H at 32 × MIC showed regular rod-shaped cells, while only cells treated with thurincin H at the elevated levels of 256 × MIC showed loss of cell integrity and rigidity. Both concentrations caused greater than 99% of cell viability reduction. In contrast, nisin caused significant cell membrane permeability at concentration as low as 2 × MIC. These results indicated a difference in the mode of action for thurincin H compared with the generalized pore-forming mechanism of many lantibiotics, such as nisin. PMID:24891232

  14. Real-Time Observation on Evolution of Droplets Morphology Affected by Electric Current Pulse in Al-Bi Immiscible Alloy

    NASA Astrophysics Data System (ADS)

    Zhu, Jing; Wang, Tongmin; Cao, Fei; Fu, Hongwang; Fu, Yanan; Xie, Honglan; Xiao, Tiqiao

    2013-05-01

    The evolution of Bi-rich droplets morphology in a solidifying Al-Bi immiscible alloy was directly observed using a synchrotron microradiography technique. The electric current pulse (ECP) was applied to control the solidification process of Al-Bi immiscible alloy. It was found that the electromagnetic pinch force and Marangoni force induced by ECP and temperature gradient, respectively, can significantly affect the distribution of Bi-rich droplets. The electromagnetic pinch force drove the droplets from the center to side; meanwhile, the Marangoni force lifted the droplets from the bottom to the top. As a result, the droplets finally distributed with a manner of "inverted triangle."

  15. A nucleolar protein that affects mating efficiency in Saccharomyces cerevisiae by altering the morphological response to pheromone.

    PubMed Central

    Kim, J; Hirsch, J P

    1998-01-01

    SSF1 and SSF2 are redundant essential yeast genes that, when overexpressed, increase the mating efficiency of cells containing a defective Ste4p Gbeta subunit. To identify the precise function of these genes in mating, different responses to pheromone were assayed in cells that either lacked or overexpressed SSF gene products. Cells containing null alleles of both SSF1 and SSF2 displayed the normal transcriptional induction response to pheromone but were unable to form mating projections. Overexpression of SSF1 conferred the ability to form mating projections on cells containing a temperature-sensitive STE4 allele, but had only a small effect on transcriptional induction. SSF1 overexpression preferentially increased the mating efficiency of a strain containing a null allele of SPA2, a gene that functions specifically in cell morphology. To investigate whether Ssf1p plays a direct physical role in mating projection formation, its subcellular location was determined. An Ssf1p-GFP fusion was found to localize to the nucleolus, implying that the role of SSF gene products in projection formation is indirect. The region of Ssf1p-GFP localization in cells undergoing projection formation was larger and more diffuse, and was often present in a specific orientation with respect to the projection. Although the function of Ssf1p appears to originate in the nucleus, it is likely that it ultimately acts on one or more of the proteins that is directly involved in the morphological response to pheromone. Because many of the proteins required for projection formation during mating are also required for bud formation during vegetative growth, regulation of the activity or amount of one or more of these proteins by Ssf1p could explain its role in both mating and dividing cells. PMID:9611192

  16. ABSENCE OF SCLEROSTIN ADVERSELY AFFECTS B CELL SURVIVAL

    PubMed Central

    Cain, Corey J.; Rueda, Randell; McLelland, Bryce; Collette, Nicole M.; Loots, Gabriela G.; Manilay, Jennifer O.

    2012-01-01

    Increased osteoblast activity in sclerostin-knockout (Sost−/−) mice results in generalized hyperostosis and bones with small bone marrow cavities due to hyperactive mineralizing osteoblast populations. Hematopoietic cell fate decisions are dependent on their local microenvironment, which contains osteoblast and stromal cell populations that support both hematopoietic stem cell quiescence and facilitate B cell development. In this study, we investigated whether high bone mass environments affect B cell development via the utilization of Sost−/− mice, a model of sclerosteosis. We found the bone marrow of Sost−/− mice to be specifically depleted of B cells, due to elevated apoptosis at all B cell developmental stages. In contrast, B cell function in the spleen was normal. Sost expression analysis confirmed that Sost is primarily expressed in osteocytes and is not expressed in any hematopoietic lineage, which indicated that the B cell defects in Sost−/− mice are non-cell autonomous and this was confirmed by transplantation of wildtype (WT) bone marrow into lethally irradiated Sost−/− recipients. WT→Sost−/− chimeras displayed a reduction in B cells, whereas reciprocal Sost−/−→WT chimeras did not, supporting the idea that the Sost−/− bone environment cannot fully support normal B cell development. Expression of the pre-B cell growth stimulating factor, Cxcl12, was significantly lower in bone marrow stromal cells of Sost−/− mice while the Wnt target genes Lef-1 and Ccnd1 remained unchanged in B cells. Taken together, these results demonstrate a novel role for Sost in the regulation of bone marrow environments that support B cells. PMID:22434688

  17. Expression level of methanol-inducible peroxisomal proteins and peroxisome morphology are affected by oxygen conditions and mitochondrial respiratory pathway function in the methylotrophic yeast Candida boidinii.

    PubMed

    Fujimura, Shuki; Yurimoto, Hiroya; Kurimoto, Shota; Matsufuji, Yoshimi; Ito, Takashi; Hayakawa, Takashi; Tomizuka, Noboru; Sakai, Yasuyoshi; Nakagawa, Tomoyuki

    2013-06-01

    In the methylotrophic yeast, Candida boidinii, methanol-inducible peroxisomal proteins, for example alcohol oxidase (AOD), dihydroxyacetone synthase (DAS), and peroxisomal glutathione peroxidase (Pmp20), were induced only under aerobic conditions, while expression of PMP47 encoding peroxisomal integral membrane protein Pmp47 was independent of oxygen conditions. Expression of the methanol-inducible peroxisomal enzymes was repressed by inhibition of the mitochondrial respiratory chain. In the respiratory-deficient (ρ0) mutant strain, their induction was at very low levels despite the presence of oxygen, whereas the expression of PMP47 was unaffected. Taken together, these facts indicate that C. boidinii can sense oxygen conditions, and that mitochondrial respiratory function may have a profound effect on induction of methanol-inducible gene expression of peroxisomal proteins. Peroxisome morphology was also affected by oxygen conditions and respiratory function. Under hypoxic conditions or respiration-inhibited conditions, cells induced by methanol contained small peroxisomes, indicating that peroxisome biogenesis and the protein import machinery were not affected by oxygen conditions but that peroxisome morphology was dependent on induction of peroxisomal matrix proteins.

  18. D-glucuronyl C5-epimerase cell type specifically affects angiogenesis pathway in different prostate cancer cells.

    PubMed

    Rosenberg, Eugenia E; Prudnikova, Tatiana Y; Zabarovsky, Eugene R; Kashuba, Vladimir I; Grigorieva, Elvira V

    2014-04-01

    D-glucuronyl C5-epimerase (GLCE) is involved in breast and lung carcinogenesis as a potential tumor suppressor gene, acting through inhibition of tumor angiogenesis and invasion/metastasis pathways. However, in prostate tumors, increased GLCE expression is associated with advanced disease, suggesting versatile effects of GLCE in different cancers. To investigate further the potential cancer-promoting effect of GLCE in prostate cancer, GLCE was ectopically re-expressed in morphologically different LNCaP and PC3 prostate cancer cells. Transcriptional profiles of normal PNT2 prostate cells, LNCaP, PC3 and DU145 prostate cancer cells, and GLCE-expressing LNCaP and PC3 cells were determined. Comparative analysis revealed the genes whose expression was changed in prostate cancer cells compared with normal PNT2 cells, and those differently expressed between the cancer cell lines (ACTA2, IL6, SERPINE1, TAGLN, SEMA3A, and CDH2). GLCE re-expression influenced mainly angiogenesis-involved genes (ANGPT1, SERPINE1, IGF1, PDGFB, TNF, IL8, TEK, IFNA1, and IFNB1) but in a cell type-specific manner (from basic deregulation of angiogenesis in LNCaP cells to significant activation in PC3 cells). Invasion/metastasis pathway was also affected (MMP1, MMP2, MMP9, S100A4, ITGA1, ITGB3, ERBB2, and FAS). The obtained results suggest activation of angiogenesis as a main molecular mechanism of pro-oncogenic effect of GLCE in prostate cancer. GLCE up-regulation plus expression pattern of a panel of six genes, discriminating morphologically different prostate cancer cell sub-types, is suggested as a potential marker of aggressive prostate cancer.

  19. Deiodinase knockdown affects zebrafish eye development at the level of gene expression, morphology and function.

    PubMed

    Houbrechts, Anne M; Vergauwen, Lucia; Bagci, Enise; Van Houcke, Jolien; Heijlen, Marjolein; Kulemeka, Bernard; Hyde, David R; Knapen, Dries; Darras, Veerle M

    2016-03-15

    Retinal development in vertebrates relies extensively on thyroid hormones. Their local availability is tightly controlled by several regulators, including deiodinases (Ds). Here we used morpholino technology to explore the roles of Ds during eye development in zebrafish. Transcriptome analysis at 3 days post fertilization (dpf) revealed a pronounced effect of knockdown of both T4-activating Ds (D1D2MO) or knockdown of T3-inactivating D3 (D3bMO) on phototransduction and retinoid recycling. This was accompanied by morphological defects (studied from 1 to 7 dpf) including reduced eye size, disturbed retinal lamination and strong reduction in rods and all four cone types. Defects were more prominent and persistent in D3-deficient fish. Finally, D3-deficient zebrafish larvae had disrupted visual function at 4 dpf and were less sensitive to a light stimulus at 5 dpf. These data demonstrate the importance of TH-activating and -inactivating Ds for correct zebrafish eye development, and point to D3b as a central player. PMID:26802877

  20. Morphological Characteristics of Maize Canopy Development as Affected by Increased Plant Density.

    PubMed

    Song, Youhong; Rui, Yukui; Bedane, Guta; Li, Jincai

    2016-01-01

    Improving crop productivity through higher plant density requires a detailed understanding of organ development in response to increased interplant competition. The objective of this paper is thus to investigate the characteristics of organ development under increased interplant competition. A field experiment was conducted to investigate organ development across 4 maize plant densities i.e. 2, 6, 12 and 20 plants m-2 (referred to PD2, PD6, PD12 and PD20 respectively). In response to increased interplant competition, lengths of both laminae and sheaths increased in lower phytomers, but decreased in upper phytomers. Sheath extension appeared to be less sensitive to increased interplant competition than lamina extension. Extension of laminae and internodes responded to increased plant density as soon as onset of mild interplant competition, but did not respond any further to severe competition. Both lamina width and internode diameter were reduced due to a smaller growth rate in response to increased plant density. Overall, this study identified that organ expansion rate can be taken as the key morphological factor to determine the degree of interplant competition.

  1. Morphological Characteristics of Maize Canopy Development as Affected by Increased Plant Density

    PubMed Central

    Song, Youhong; Rui, Yukui; Bedane, Guta; Li, Jincai

    2016-01-01

    Improving crop productivity through higher plant density requires a detailed understanding of organ development in response to increased interplant competition. The objective of this paper is thus to investigate the characteristics of organ development under increased interplant competition. A field experiment was conducted to investigate organ development across 4 maize plant densities i.e. 2, 6, 12 and 20 plants m–2 (referred to PD2, PD6, PD12 and PD20 respectively). In response to increased interplant competition, lengths of both laminae and sheaths increased in lower phytomers, but decreased in upper phytomers. Sheath extension appeared to be less sensitive to increased interplant competition than lamina extension. Extension of laminae and internodes responded to increased plant density as soon as onset of mild interplant competition, but did not respond any further to severe competition. Both lamina width and internode diameter were reduced due to a smaller growth rate in response to increased plant density. Overall, this study identified that organ expansion rate can be taken as the key morphological factor to determine the degree of interplant competition. PMID:27129101

  2. The influence of morphology on charge transport/recombination dynamics in planar perovskite solar cells

    NASA Astrophysics Data System (ADS)

    Yu, Man; Wang, Yi; Wang, Hao-Yi; Han, Jun; Qin, Yujun; Zhang, Jian-Ping; Ai, Xi-Cheng

    2016-10-01

    The photovoltaic performance of planar perovskite solar cell is significantly influenced by the morphology of perovskite film. In this work, five kinds of devices with different perovskite film morphologies were prepared by varying the concentration of CH3NH3Cl in precursor solutions. We found that best morphology of perovskite film results in the excellent photovoltaic performance with an average efficiency of 15.52% and a champion efficiency of 16.38%. Transient photovoltage and photocurrent measurements are performed to elucidate the mechanism of photoelectric conversion processes, which shows that the charge recombination is effectively suppressed and the charge transport is obviously promoted by optimized morphology.

  3. Reciprocity in predator-prey interactions: exposure to defended prey and predation risk affects intermediate predator life history and morphology.

    PubMed

    Hammill, Edd; Beckerman, Andrew P

    2010-05-01

    A vast body of literature exists documenting the morphological, behavioural and life history changes that predators induce in prey. However, little attention has been paid to how these induced changes feed back and affect the predators' life history and morphology. Larvae of the phantom midge Chaoborus flavicans are intermediate predators in a food web with Daphnia pulex as the basal resource and planktivorous fish as the top predator. C. flavicans prey on D. pulex and are themselves prey for fish; as D. pulex induce morphological defences in the presence of C. flavicans this is an ideal system in which to evaluate the effects of defended prey and top predators on an intermediate consumer. We assessed the impact on C. flavicans life history and morphology of foraging on defended prey while also being exposed to the non-lethal presence of a top fish predator. We tested the basic hypothesis that the effects of defended prey will depend on the presence or absence of top predator predation risk. Feeding rate was significantly reduced and time to pupation was significantly increased by defended morph prey. Gut size, development time, fecundity, egg size and reproductive effort respond to fish chemical cues directly or significantly alter the relationship between a trait and body size. We found no significant interactions between prey morph and the non-lethal presence of a top predator, suggesting that the effects of these two biological factors were additive or singularly independent. Overall it appears that C. flavicans is able to substantially modify several aspects of its biology, and while some changes appear mere consequences of resource limitation others appear facultative in nature. PMID:19936795

  4. Improved performance by morphology control via fullerenes in PBDT-TBT-alkoBT based organic solar cells

    SciTech Connect

    Khatiwada, Devendra; Venkatesan, Swaminathan; Chen, QIliang; Chen, Jihua; Adhikari, Nirmal; Dubey, Ashish; Mitul, Abu Farzan; Mohammed, Lal; Qiao, Qiquan

    2015-07-03

    In this work, we report improved performance by controlling morphology using different fullerene derivatives in poly{2-octyldodecyloxy-benzo[1,2-b;3,4-b]dithiophene-alt-5,6-bis(dodecyloxy)-4,7- di(thieno[3,2-b]thiophen-2-yl)-benzo[c][1,2,5]thiadiazole} (PBDT-TBT-alkoBT) based organic solar cells. PC60BM and PC70BM fullerenes were used to investigate the characteristic change in morphology and device performance. Fullerene affects device efficiency by changing active layer morphology. PC70BM with broader absorption than PC60BM resulted in reduced device performance which was elucidated by the intermixed granular morphology separating each larger grain in the PC70BM/polymer composite layer which created higher density of traps. However after adding additive 1,8-diiodooctane (DIO), the fibrous morphology was observed due to reduced solubility of polymer and increased solubility of PC70BM in chloroform. The fibrous morphology improved charge transport leading to increase in overall device performance. Atomic force microscopies (AFM), photo induced charge extraction by linearly increasing voltage (photo-CELIV), and Kelvin prove force microscope (KPFM) were used to investigate nanoscale morphology of active layer with different fullerene derivatives. For PC60BM based active layer, AFM images revealed dense fibrous morphology and more distinct fibrous morphology was observed by adding DIO. The PC70BM based active layer only exhibited intermixed granular morphology instead of fibrous morphology observed in PC60BM based active layer. However, addition of DIO in PC70BM based active layer led to fibrous morphology. When additive DIO was not used, a wider distribution of surface potential was observed for PC70BM than PC60BM based active layer by KPFM measurements, indicating 2 polymer and fullerene domains are separated. When DIO was used, narrower distribution of surface potential for both PC70

  5. Improved performance by morphology control via fullerenes in PBDT-TBT-alkoBT based organic solar cells

    DOE PAGES

    Khatiwada, Devendra; Venkatesan, Swaminathan; Chen, QIliang; Chen, Jihua; Adhikari, Nirmal; Dubey, Ashish; Mitul, Abu Farzan; Mohammed, Lal; Qiao, Qiquan

    2015-07-03

    In this work, we report improved performance by controlling morphology using different fullerene derivatives in poly{2-octyldodecyloxy-benzo[1,2-b;3,4-b]dithiophene-alt-5,6-bis(dodecyloxy)-4,7- di(thieno[3,2-b]thiophen-2-yl)-benzo[c][1,2,5]thiadiazole} (PBDT-TBT-alkoBT) based organic solar cells. PC60BM and PC70BM fullerenes were used to investigate the characteristic change in morphology and device performance. Fullerene affects device efficiency by changing active layer morphology. PC70BM with broader absorption than PC60BM resulted in reduced device performance which was elucidated by the intermixed granular morphology separating each larger grain in the PC70BM/polymer composite layer which created higher density of traps. However after adding additive 1,8-diiodooctane (DIO), the fibrous morphology was observed due to reduced solubility of polymer and increasedmore » solubility of PC70BM in chloroform. The fibrous morphology improved charge transport leading to increase in overall device performance. Atomic force microscopies (AFM), photo induced charge extraction by linearly increasing voltage (photo-CELIV), and Kelvin prove force microscope (KPFM) were used to investigate nanoscale morphology of active layer with different fullerene derivatives. For PC60BM based active layer, AFM images revealed dense fibrous morphology and more distinct fibrous morphology was observed by adding DIO. The PC70BM based active layer only exhibited intermixed granular morphology instead of fibrous morphology observed in PC60BM based active layer. However, addition of DIO in PC70BM based active layer led to fibrous morphology. When additive DIO was not used, a wider distribution of surface potential was observed for PC70BM than PC60BM based active layer by KPFM measurements, indicating 2 polymer and fullerene domains are separated. When DIO was used, narrower distribution of surface potential for both PC70BM and PC60BM based active layers was observed. Photo-CELIV experiment

  6. Thermal stabilisation of polymer-fullerene bulk heterojunction morphology for efficient photovoltaic solar cells.

    PubMed

    Derue, Lionel; Dautel, Olivier; Tournebize, Aurélien; Drees, Martin; Pan, Hualong; Berthumeyrie, Sébastien; Pavageau, Bertrand; Cloutet, Eric; Chambon, Sylvain; Hirsch, Lionel; Rivaton, Agnès; Hudhomme, Piétrick; Facchetti, Antonio; Wantz, Guillaume

    2014-09-01

    A novel stable bisazide molecule that can freeze the bulk heterojunction morphology at its optimized layout by specifically bonding to fullerenes is reported. The concept is demonstrated with various polymers: fullerene derivatives systems enable highly thermally stable polymer solar cells.

  7. Gamma radiation affects active electrolyte transport by rabbit ileum. II. Correlation of alanine and theophylline response with morphology

    SciTech Connect

    Gunter-Smith, P.J.

    1989-03-01

    The response of ileal segments isolated from rabbits to an actively transported amino acid and a secretagogue was evaluated following exposure to 10 Gy whole-body gamma irradiation. The ability of ileal segments to respond to the actively transported amino acid, alanine, was not significantly diminished until 96 h postexposure. Decreased responsiveness to the secretagogue, theophylline, occurred earlier at 72 h. These effects did not appear to be accounted for by decreased food intake of irradiated animals alone. Examination of intestinal morphological changes with respect to these changes in electrolyte transport revealed that decreased amino acid transport coincides with loss of intestinal villi. Although a morphological correlate of decreased secretory response was not as striking as that for absorption, the theophylline response appeared to decline concomitant with the appearance of increased mitotic activity in the intestinal crypts. The results of this study indicate that, following a dose of 10 Gy, the inability of these tissues to respond to amino acids is due to a loss of mature villus absorptive cells subsequent to denudation of the intestinal mucosa. There appeared to be little impairment of cell membrane transport processes for alanine. In contrast, the decreased secretory response could not be correlated with the disappearance of any one cell type and perhaps results from increased proliferation in the crypts at the expense of differentiation.

  8. Gamma-radiation affects active electrolyte transport by rabbit ileum. 2. Correlation of alanine and theophylline response with morphology

    SciTech Connect

    Gunter-Smith, P.J.

    1989-01-01

    The response of ileal segments isolated from rabbits to an actively transported amino acid and a secretagogue was evaluated following exposure to 10-Gy whole-body gamma irradiation. The ability of ileal segments to respond to the actively transported amino acid, alanine, was not significantly diminished until 96 h postexposure. Decreased responsiveness to the secretagogue, theophylline, occurred earlier at 72 h. These effects did not appear to be accounted for by decreased food intake of irradiated animals alone. Examination of intestinal morphological changes with respect to these changes in electrolyte transport revealed that decreased amino acid transport coincides with loss of intestinal villi. Although a morphological correlate of decreased secretory response was not as striking as that for absorption, the theophylline response appeared to decline concomitant with the appearance of increased mitotic activity in the intestinal crypts. The result of this study indicate that, following a dose of 10 Gy, the inability of these tissues to respond to amino acids is due to a loss of mature villus absorptive cells subsequent to denudation of the intestinal mucosa. There appeared to be little impairment of cell membrane transport processes for alanine. In contrast, the decreased secretory response could not be correlated with the disappearance of any one cell type and perhaps results from increased proliferation in the crypts at the expense of differentiation.

  9. Endothelial Cell Morphology and Migration are Altered by Changes in Gravitational Fields

    NASA Technical Reports Server (NTRS)

    Melhado, Caroline; Sanford, Gary; Harris-Hooker, Sandra

    1997-01-01

    Many of the physiological changes of the cardiovascular system during space flight may originate from the dysfunction of basic biological mechanisms caused by microgravity. The weightlessness affects the system when blood and other fluids move to the upper body causing the heart to enlarge to handle the increased blood flow to the upper extremities and decrease circulating volume. Increase arterial pressure triggers baroreceptors which signal the brain to adjust heart rate. Hemodynarnic studies indicate that the microgravity-induced headward fluid redistribution results in various cardiovascular changes such as; alteration of vascular permeability resulting in lipid accumulation in the lumen of the vasculature and degeneration of the the vascular wall, capillary alteration with extensive endothelial invagination. Achieving a true microgravity environment in ground based studies for prolonged periods is virtually impossible. The application of vector-averaged gravity to mammalian cells using horizontal clinostat produces alterations of cellular behavior similar to those observed in microgravity. Similarly, the low shear, horizontally rotating bioreactor (originally designed by NASA) also duplicates several properties of microgravity. Additionally, increasing gravity, i.e., hypcrgravity is easily achieved. Hypergravity has been found to increase the proliferation of several different cell lines (e.g., chick embryo fibroblasts) while decreasing cell motility and slowing liver regeneration following partial hepatectomy. The effect of altered gravity on cells maybe similar to those of other physical forces, i.e. shear stress. Previous studies examining laminar flow and shear stress on endothelial cells found that the cells elongate, orient with the direction of flow, and reorganize their F-actin structure, with concomitant increase in cell stiffness. These studies suggest that alterations in the gravity environment will change the behavior of most cells, including

  10. Effects of risedronate on the morphology and viability of gingiva-derived mesenchymal stem cells

    PubMed Central

    KIM, BO-BAE; KO, YOUNGKYUNG; PARK, JUN-BEOM

    2015-01-01

    Risedronate has been used for the prevention and treatment of postmenopausal and corticosteroid-induced osteoporosis. The present study was performed to evaluate the effects of risedronate on the morphology and viability of human stem cells derived from the gingiva. Stem cells derived from the gingiva were grown in the presence of risedronate at concentrations that ranged from 1 to 10 µM. The morphology of the cells was viewed under an inverted microscope, and cell proliferation was analyzed with a cell counting kit-8 (CCK-8) on days 2, 4 and 7. The untreated control group showed a spindle-shaped, fibroblast-like morphology. The shapes of the cells treated with 1 and 5 µM risedronate were similar to that of the control group on day 2. However, morphology of the 10 µM group markedly differed from that of the control group. The shapes of the cells in the 1, 5 and 10 µM groups were rounder, and pronounced alterations when compared with the untreated control group were noted in all groups on day 7. The cultures growing in the presence of risedronate showed decreased CCK-8 values on day 7. In conclusion, risedronate produced notable alterations in the morphology of the cells and reduced the viability of gingival mesenchymal stem cells. PMID:26623028

  11. Changes, and the Relevance Thereof, in Mitochondrial Morphology during Differentiation into Endothelial Cells.

    PubMed

    Shin, Ji Won; Park, So Hee; Kang, Yun Gyeong; Wu, Yanru; Choi, Hyun Ju; Shin, Jung-Woog

    2016-01-01

    The roles of mitochondria in various physiological functions of vascular endothelial cells have been investigated extensively. Morphological studies in relation to physiological functions have been performed. However, there have been few reports of morphological investigations related to stem cell differentiation. This was the first morphological study of mitochondria in relation to endothelial differentiation and focused on quantitative analysis of changes in mitochondrial morphology, number, area, and length during differentiation of human mesenchymal stem cells (hMSCs) into endothelial-like cells. To induce differentiation, we engaged vascular endothelial growth factors and flow-induced shear stress. Cells were classified according to the expression of von Willebrand factor as hMSCs, differentiating cells, and almost fully differentiated cells. Based on imaging analysis, we investigated changes in mitochondrial number, area, and length. In addition, mitochondrial networks were quantified on a single-mitochondrion basis by introducing a branch form factor. The data indicated that the mitochondrial number, area per cell, and length were decreased with differentiation. The mitochondrial morphology became simpler with progression of differentiation. These findings could be explained in view of energy level during differentiation; a higher level of energy is needed during differentiation, with larger numbers of mitochondria with branches. Application of this method to differentiation into other lineages will explain the energy levels required to control stem cell differentiation. PMID:27517609

  12. Changes, and the Relevance Thereof, in Mitochondrial Morphology during Differentiation into Endothelial Cells

    PubMed Central

    Shin, Ji Won; Park, So Hee; Kang, Yun Gyeong; Wu, Yanru; Choi, Hyun Ju

    2016-01-01

    The roles of mitochondria in various physiological functions of vascular endothelial cells have been investigated extensively. Morphological studies in relation to physiological functions have been performed. However, there have been few reports of morphological investigations related to stem cell differentiation. This was the first morphological study of mitochondria in relation to endothelial differentiation and focused on quantitative analysis of changes in mitochondrial morphology, number, area, and length during differentiation of human mesenchymal stem cells (hMSCs) into endothelial-like cells. To induce differentiation, we engaged vascular endothelial growth factors and flow-induced shear stress. Cells were classified according to the expression of von Willebrand factor as hMSCs, differentiating cells, and almost fully differentiated cells. Based on imaging analysis, we investigated changes in mitochondrial number, area, and length. In addition, mitochondrial networks were quantified on a single-mitochondrion basis by introducing a branch form factor. The data indicated that the mitochondrial number, area per cell, and length were decreased with differentiation. The mitochondrial morphology became simpler with progression of differentiation. These findings could be explained in view of energy level during differentiation; a higher level of energy is needed during differentiation, with larger numbers of mitochondria with branches. Application of this method to differentiation into other lineages will explain the energy levels required to control stem cell differentiation. PMID:27517609

  13. Functional differences between two morphologically distinct cell subpopulations within a human colorectal carcinoma cell line.

    PubMed

    Solimene, A C; Carneiro, C R; Melati, I; Lopes, J D

    2001-05-01

    The LISP-I human colorectal adenocarcinoma cell line was isolated from a hepatic metastasis at the Ludwig Institute, São Paulo, SP, Brazil. The objective of the present study was to isolate morphologically different subpopulations within the LISP-I cell line, and characterize some of their behavioral aspects such as adhesion to and migration towards extracellular matrix components, expression of intercellular adhesion molecules and tumorigenicity in vitro. Once isolated, the subpopulations were submitted to adhesion and migration assays on laminin and fibronectin (crucial proteins to invasion and metastasis), as well as to anchorage-independent growth. Two morphologically different subpopulations were isolated: LISP-A10 and LISP-E11. LISP-A10 presents a differentiated epithelial pattern, and LISP-E11 is fibroblastoid, suggesting a poorly differentiated pattern. LISP-A10 expressed the two intercellular adhesion molecules tested, carcinoembryonic antigen (CEA) and desmoglein, while LISP-E11 expressed only low amounts of CEA. On the other hand, adhesion to laminin and fibronectin as well as migration towards these extracellular matrix proteins were higher in LISP-E11, as expected from its poorly differentiated phenotype. Both subpopulations showed anchorage-independent growth on a semi-solid substrate. These results raise the possibility that the heterogeneity found in the LISP-I cell line, which might have contributed to its ability to metastasize, was due to at least two different subpopulations herein identified. PMID:11323753

  14. Mapping Quantitative Trait Loci Affecting Biochemical and Morphological Fruit Properties in Eggplant (Solanum melongena L.).

    PubMed

    Toppino, Laura; Barchi, Lorenzo; Lo Scalzo, Roberto; Palazzolo, Eristanna; Francese, Gianluca; Fibiani, Marta; D'Alessandro, Antonietta; Papa, Vincenza; Laudicina, Vito A; Sabatino, Leo; Pulcini, Laura; Sala, Tea; Acciarri, Nazzareno; Portis, Ezio; Lanteri, Sergio; Mennella, Giuseppe; Rotino, Giuseppe L

    2016-01-01

    Eggplant berries are a source of health-promoting metabolites including antioxidant and nutraceutical compounds, mainly anthocyanins and chlorogenic acid; however, they also contain some anti-nutritional compounds such as steroidal glycoalkaloids (SGA) and saponins, which are responsible for the bitter taste of the flesh and with potential toxic effects on humans. Up to now, Quantitative Trait Loci (QTL) for the metabolic content are far from being characterized in eggplant, thus hampering the application of breeding programs aimed at improving its fruit quality. Here we report on the identification of some QTL for the fruit metabolic content in an F2 intraspecific mapping population of 156 individuals, obtained by crossing the eggplant breeding lines "305E40" × "67/3." The same population was previously employed for the development of a RAD-tag based linkage map and the identification of QTL associated to morphological and physiological traits. The mapping population was biochemically characterized for both fruit basic qualitative data, like dry matter, °Brix, sugars, and organic acids, as well as for health-related compounds such chlorogenic acid, (the main flesh monomeric phenol), the two peel anthocyanins [i.e., delphinidin-3-rutinoside (D3R) and delphinidin-3-(p- coumaroylrutinoside)-5-glucoside (nasunin)] and the two main steroidal glycoalkaloids, solasonine, and solamargine. For most of the traits, one major QTL (PVE ≥10%) was spotted and putative orthologies with other Solanaceae crops are discussed. The present results supply valuable information to eggplant breeders on the inheritance of key fruit quality traits, thus providing potential tools to assist future breeding programs. PMID:26973692

  15. Mapping Quantitative Trait Loci Affecting Biochemical and Morphological Fruit Properties in Eggplant (Solanum melongena L.).

    PubMed

    Toppino, Laura; Barchi, Lorenzo; Lo Scalzo, Roberto; Palazzolo, Eristanna; Francese, Gianluca; Fibiani, Marta; D'Alessandro, Antonietta; Papa, Vincenza; Laudicina, Vito A; Sabatino, Leo; Pulcini, Laura; Sala, Tea; Acciarri, Nazzareno; Portis, Ezio; Lanteri, Sergio; Mennella, Giuseppe; Rotino, Giuseppe L

    2016-01-01

    Eggplant berries are a source of health-promoting metabolites including antioxidant and nutraceutical compounds, mainly anthocyanins and chlorogenic acid; however, they also contain some anti-nutritional compounds such as steroidal glycoalkaloids (SGA) and saponins, which are responsible for the bitter taste of the flesh and with potential toxic effects on humans. Up to now, Quantitative Trait Loci (QTL) for the metabolic content are far from being characterized in eggplant, thus hampering the application of breeding programs aimed at improving its fruit quality. Here we report on the identification of some QTL for the fruit metabolic content in an F2 intraspecific mapping population of 156 individuals, obtained by crossing the eggplant breeding lines "305E40" × "67/3." The same population was previously employed for the development of a RAD-tag based linkage map and the identification of QTL associated to morphological and physiological traits. The mapping population was biochemically characterized for both fruit basic qualitative data, like dry matter, °Brix, sugars, and organic acids, as well as for health-related compounds such chlorogenic acid, (the main flesh monomeric phenol), the two peel anthocyanins [i.e., delphinidin-3-rutinoside (D3R) and delphinidin-3-(p- coumaroylrutinoside)-5-glucoside (nasunin)] and the two main steroidal glycoalkaloids, solasonine, and solamargine. For most of the traits, one major QTL (PVE ≥10%) was spotted and putative orthologies with other Solanaceae crops are discussed. The present results supply valuable information to eggplant breeders on the inheritance of key fruit quality traits, thus providing potential tools to assist future breeding programs.

  16. Activation and aponeurosis morphology affect in vivo muscle tissue strains near the myotendinous junction.

    PubMed

    Fiorentino, Niccolo M; Epstein, Frederick H; Blemker, Silvia S

    2012-02-23

    Hamstring strain injury is one of the most common injuries in athletes, particularly for sports that involve high speed running. The aims of this study were to determine whether muscle activation and internal morphology influence in vivo muscle behavior and strain injury susceptibility. We measured tissue displacement and strains in the hamstring muscle injured most often, the biceps femoris long head muscle (BFLH), using cine DENSE dynamic magnetic resonance imaging. Strain measurements were used to test whether strain magnitudes are (i) larger during active lengthening than during passive lengthening and (ii) larger for subjects with a relatively narrow proximal aponeurosis than a wide proximal aponeurosis. Displacement color maps showed higher tissue displacement with increasing lateral distance from the proximal aponeurosis for both active lengthening and passive lengthening, and higher tissue displacement for active lengthening than passive lengthening. First principal strain magnitudes were averaged in a 1cm region near the myotendinous junction, where injury is most frequently observed. It was found that strains are significantly larger during active lengthening (0.19 SD 0.09) than passive lengthening (0.13 SD 0.06) (p<0.05), which suggests that elevated localized strains may be a mechanism for increased injury risk during active as opposed to passive lengthening. First principal strains were higher for subjects with a relatively narrow aponeurosis width (0.26 SD 0.15) than wide (0.14 SD 0.04) (p<0.05). This result suggests that athletes who have BFLH muscles with narrow proximal aponeuroses may have an increased risk for BFLH strain injuries.

  17. Mapping Quantitative Trait Loci Affecting Biochemical and Morphological Fruit Properties in Eggplant (Solanum melongena L.)

    PubMed Central

    Toppino, Laura; Barchi, Lorenzo; Lo Scalzo, Roberto; Palazzolo, Eristanna; Francese, Gianluca; Fibiani, Marta; D'Alessandro, Antonietta; Papa, Vincenza; Laudicina, Vito A.; Sabatino, Leo; Pulcini, Laura; Sala, Tea; Acciarri, Nazzareno; Portis, Ezio; Lanteri, Sergio; Mennella, Giuseppe; Rotino, Giuseppe L.

    2016-01-01

    Eggplant berries are a source of health-promoting metabolites including antioxidant and nutraceutical compounds, mainly anthocyanins and chlorogenic acid; however, they also contain some anti-nutritional compounds such as steroidal glycoalkaloids (SGA) and saponins, which are responsible for the bitter taste of the flesh and with potential toxic effects on humans. Up to now, Quantitative Trait Loci (QTL) for the metabolic content are far from being characterized in eggplant, thus hampering the application of breeding programs aimed at improving its fruit quality. Here we report on the identification of some QTL for the fruit metabolic content in an F2 intraspecific mapping population of 156 individuals, obtained by crossing the eggplant breeding lines “305E40” × “67/3.” The same population was previously employed for the development of a RAD-tag based linkage map and the identification of QTL associated to morphological and physiological traits. The mapping population was biochemically characterized for both fruit basic qualitative data, like dry matter, °Brix, sugars, and organic acids, as well as for health-related compounds such chlorogenic acid, (the main flesh monomeric phenol), the two peel anthocyanins [i.e., delphinidin-3-rutinoside (D3R) and delphinidin-3-(p- coumaroylrutinoside)-5-glucoside (nasunin)] and the two main steroidal glycoalkaloids, solasonine, and solamargine. For most of the traits, one major QTL (PVE ≥10%) was spotted and putative orthologies with other Solanaceae crops are discussed. The present results supply valuable information to eggplant breeders on the inheritance of key fruit quality traits, thus providing potential tools to assist future breeding programs. PMID:26973692

  18. Accumulation of distinct prelamin A variants in human diploid fibroblasts differentially affects cell homeostasis

    SciTech Connect

    Candelario, Jose; Borrego, Stacey; Reddy, Sita; Comai, Lucio

    2011-02-01

    Lamin A is a component of the nuclear lamina that plays a major role in the structural organization and function of the nucleus. Lamin A is synthesized as a prelamin A precursor which undergoes four sequential post-translational modifications to generate mature lamin A. Significantly, a large number of point mutations in the LMNA gene cause a range of distinct human disorders collectively known as laminopathies. The mechanisms by which mutations in lamin A affect cell function and cause disease are unclear. Interestingly, recent studies have suggested that alterations in the normal lamin A pathway can contribute to cellular dysfunction. Specifically, we and others have shown, at the cellular level, that in the absence of mutations or altered splicing events, increased expression of wild-type prelamin A results in a growth defective phenotype that resembles that of cells expressing the mutant form of lamin A, termed progerin, associated with Hutchinson-Gilford Progeria syndrome (HGPS). Remarkably, the phenotypes of cells expressing elevated levels of wild-type prelamin A can be reversed by either treatment with farnesyltransferase inhibitors or overexpression of ZMPSTE24, a critical prelamin A processing enzyme, suggesting that minor increases in the steady-state levels of one or more prelamin A intermediates is sufficient to induce cellular toxicity. Here, to investigate the molecular basis of the lamin A pathway toxicity, we characterized the phenotypic changes occurring in cells expressing distinct prelamin A variants mimicking specific prelamin A processing intermediates. This analysis demonstrates that distinct prelamin A variants differentially affect cell growth, nuclear membrane morphology, nuclear distribution of lamin A and the fundamental process of transcription. Expression of prelamin A variants that are constitutively farnesylated induced the formation of lamin A aggregates and dramatic changes in nuclear membrane morphology, which led to reduced

  19. Modeling the Excess Cell Surface Stored in a Complex Morphology of Bleb-Like Protrusions

    PubMed Central

    Wessler, Timothy; Yang, Xiaofeng; Chen, Alex; Roach, Nathan; Elston, Timothy C.; Wang, Qi; Jacobson, Ken; Forest, M. Gregory

    2016-01-01

    Cells transition from spread to rounded morphologies in diverse physiological contexts including mitosis and mesenchymal-to-amoeboid transitions. When these drastic shape changes occur rapidly, cell volume and surface area are approximately conserved. Consequently, the rounded cells are suddenly presented with a several-fold excess of cell surface whose area far exceeds that of a smooth sphere enclosing the cell volume. This excess is stored in a population of bleb-like protrusions (BLiPs), whose size distribution is shown by electron micrographs to be skewed. We introduce three complementary models of rounded cell morphologies with a prescribed excess surface area. A 2D Hamiltonian model provides a mechanistic description of how discrete attachment points between the cell surface and cortex together with surface bending energy can generate a morphology that satisfies a prescribed excess area and BLiP number density. A 3D random seed-and-growth model simulates efficient packing of BLiPs over a primary rounded shape, demonstrating a pathway for skewed BLiP size distributions that recapitulate 3D morphologies. Finally, a phase field model (2D and 3D) posits energy-based constitutive laws for the cell membrane, nematic F-actin cortex, interior cytosol, and external aqueous medium. The cell surface is equipped with a spontaneous curvature function, a proxy for the cell surface-cortex couple, that is a priori unknown, which the model “learns” from the thin section transmission electron micrograph image (2D) or the “seed and growth” model image (3D). Converged phase field simulations predict self-consistent amplitudes and spatial localization of pressure and stress throughout the cell for any posited stationary morphology target and cell compartment constitutive properties. The models form a general framework for future studies of cell morphological dynamics in a variety of biological contexts. PMID:27015526

  20. VMP1-deficient Chlamydomonas exhibits severely aberrant cell morphology and disrupted cytokinesis

    PubMed Central

    2014-01-01

    Background The versatile Vacuole Membrane Protein 1 (VMP1) has been previously investigated in six species. It has been shown to be essential in macroautophagy, where it takes part in autophagy initiation. In addition, VMP1 has been implicated in organellar biogenesis; endo-, exo- and phagocytosis, and protein secretion; apoptosis; and cell adhesion. These roles underly its proven involvement in pancreatitis, diabetes and cancer in humans. Results In this study we analyzed a VMP1 homologue from the green alga Chlamydomonas reinhardtii. CrVMP1 knockdown lines showed severe phenotypes, mainly affecting cell division as well as the morphology of cells and organelles. We also provide several pieces of evidence for its involvement in macroautophagy. Conclusion Our study adds a novel role to VMP1's repertoire, namely the regulation of cytokinesis. Though the directness of the observed effects and the mechanisms underlying them remain to be defined, the protein's involvement in macroautophagy in Chlamydomonas, as found by us, suggests that CrVMP1 shares molecular characteristics with its animal and protist counterparts. PMID:24885763

  1. Adhesion and morphology of fibroblastic cells cultured on different polymeric biomaterials.

    PubMed

    Lombello, C B; Santos, A R; Malmonge, S M; Barbanti, S H; Wada, M L F; Duek, E A R

    2002-09-01

    Cell adhesion is influenced by the physical and chemical characteristics of the materials used as substrate for cell culturing. In this work, we evaluated the influence of the morphological and chemical characteristics of different polymeric substrates on the adhesion and morphology of fibroblastic cells. Cell growth on poly (L-lactic acid) [PLLA] membranes and poly(2-hydroxy ethyl methacrylate) [polyHEMA], poly(2-hydroxy ethyl methacrylate)-cellulose acetate [polyHEMA-CA] and poly(2-hydroxy ethyl methacrylate)-poly(methyl methacrylate-co-acrylic acid) [polyHEMA-poly(MMA-co-AA)] hydrogels of different densities and pore diameters was examined. Cells adhered preferentially to more negatively charged substrates, with polyHEMA hydrogels being more adhesive than the other substractes. The pores present in PLLA membranes did not interfere with adhesion, but the cells showed a distinctive morphology on each membrane.

  2. Morphology, Growth, and Size Limit of Bacterial Cells

    NASA Astrophysics Data System (ADS)

    Jiang, Hongyuan; Sun, Sean X.

    2010-07-01

    Bacterial cells utilize a living peptidoglycan network (PG) to separate the cell interior from the surroundings. The shape of the cell is controlled by PG synthesis and cytoskeletal proteins that form bundles and filaments underneath the cell wall. The PG layer also resists turgor pressure and protects the cell from osmotic shock. We argue that mechanical influences alter the chemical equilibrium of the reversible PG assembly and determine the cell shape and cell size. Using a mechanochemical approach, we show that the cell shape can be regarded as a steady state of a growing network under the influence of turgor pressure and mechanical stress. Using simple elastic models, we predict the size of common spherical and rodlike bacteria. The influence of cytoskeletal bundles such as crescentin and MreB are discussed within the context of our model.

  3. [MORPHOLOGICAL FEATURES OF RAT MUCOUS MEMBRANE OF THE TONGUE EARLY AFFECTED BY ACRYLIC RESIN MONOMER].

    PubMed

    Davydenko, V; Nidzelskiy, M; Starchenko, I; Davydenko, A; Kuznetsov, V

    2016-03-01

    Base materials, made on the basis of various derivatives of acrylic and methacrylic acids, have been widely used in prosthetic dentistry. Free monomer, affecting the tissues of prosthetic bed and the whole body, is always found in dentures. Therefore, study of the effect of acrylic resins' monomer on mucous membrane of the tongue is crucial. Rat tongue is very similar to human tongue, and this fact has become the basis for selecting these animals to be involved into the experiment. The paper presents the findings related to the effect of "Ftoraks" base acrylic resin monomer on the state of rat mucous membrane of the tongue and its regeneration. The microscopy has found that the greatest changes in the mucous membrane of the tongue occur on day 3 and 7 day after applying the monomer and are of erosive and inflammatory nature. Regeneration of tongue epithelium slows down. PMID:27119844

  4. Morphology of human embryonic kidney cells in culture after space flight

    NASA Technical Reports Server (NTRS)

    Todd, P.; Kunze, M. E.; Williams, K.; Morrison, D. R.; Lewis, M. L.; Barlow, G. H.

    1985-01-01

    The ability of human embyronic kidney cells to differentiate into small epithelioid, large epithelioid, domed, and fenestrated morphological cell types following space flight is examined. Kidney cells exposed to 1 day at 1 g, then 1 day in orbit, and a 12 minute passage through the electrophoretic separator are compared with control cultures. The data reveal that 70 percent of small epithelioid, 16 percent of large epithelioid, 9 percent of dome-forming, and 5 percent of fenestrated cells formed in the space exposed cells; the distributions correlate well with control data. The formation of domed cells from cells cultured from low electrophoretic mobility fractions and small epithelioid cells from high mobility fractions is unaffected by space flight conditions. It is concluded that storage under microgravity conditions does not influence the morphological differentiation of human embryonic kidney cells in low-passage culture.

  5. Comparison of Morphological and Functional Endothelial Cell Changes after Cataract Surgery: Phacoemulsification Versus Manual Small-Incision Cataract Surgery

    PubMed Central

    Ganekal, Sunil; Nagarajappa, Ashwini

    2014-01-01

    Purpose: To compare the morphological (cell density, coefficient of variation and standard deviation) and functional (central corneal thickness) endothelial changes after phacoemulsification versus manual small-incision cataract surgery (MSICS). Design: Prospective randomized control study. Materials and Methods: In this prospective randomized control study, patients were randomly allocated to undergo phacoemulsification (Group 1, n = 100) or MSICS (Group 2, n = 100) using a random number Table. The patients underwent complete ophthalmic evaluation and specular microscopy preoperatively and at 1and 6 weeks postoperatively. Functional and morphological endothelial evaluation was Noncon ROBO PACHY SP-9000 specular microscope. Phacoemulsification was performed, the chop technique and MSICS, by the viscoexpression technique. Results: The mean difference in central corneal thickness at baseline and 1 week between Group 1 and Group 2 was statistically significant (P = 0.027). However, this difference at baseline when compared to 6 week and 1 week, 6 weeks was not statistically significant (P > 0.05). The difference in mean endothelial cell density between groups at 1 week and 6 weeks was statistically significant (P = 0.016). The mean coefficient of variation and mean standard deviation between groups were not statistically significant (P > 0.05, both comparisons). Conclusion: The central corneal thickness, coefficient of variation, and standard deviation were maintained in both groups indicating that the function and morphology of endothelial cells was not affected despite an initial reduction in endothelial cell number in MSICS. Thus, MSICS remains a safe option in the developing world. PMID:24669147

  6. Effect of cold plasma on glial cell morphology studied by atomic force microscopy.

    PubMed

    Recek, Nina; Cheng, Xiaoqian; Keidar, Michael; Cvelbar, Uros; Vesel, Alenka; Mozetic, Miran; Sherman, Jonathan

    2015-01-01

    The atomic force microscope (AFM) is broadly used to study the morphology of cells. The morphological characteristics and differences of the cell membrane between normal human astrocytes and glial tumor cells are not well explored. Following treatment with cold atmospheric plasma, evaluation of the selective effect of plasma on cell viability of tumor cells is poorly understood and requires further evaluation. Using AFM we imaged morphology of glial cells before and after cold atmospheric plasma treatment. To look more closely at the effect of plasma on cell membrane, high resolution imaging was used. We report the differences between normal human astrocytes and human glioblastoma cells by considering the membrane surface details. Our data, obtained for the first time on these cells using atomic force microscopy, argue for an architectural feature on the cell membrane, i.e. brush layers, different in normal human astrocytes as compared to glioblastoma cells. The brush layer disappears from the cell membrane surface of normal E6/E7 cells and is maintained in the glioblastoma U87 cells after plasma treatment.

  7. Effect of Cold Plasma on Glial Cell Morphology Studied by Atomic Force Microscopy

    PubMed Central

    Recek, Nina; Cheng, Xiaoqian; Keidar, Michael; Cvelbar, Uros; Vesel, Alenka; Mozetic, Miran; Sherman, Jonathan

    2015-01-01

    The atomic force microscope (AFM) is broadly used to study the morphology of cells. The morphological characteristics and differences of the cell membrane between normal human astrocytes and glial tumor cells are not well explored. Following treatment with cold atmospheric plasma, evaluation of the selective effect of plasma on cell viability of tumor cells is poorly understood and requires further evaluation. Using AFM we imaged morphology of glial cells before and after cold atmospheric plasma treatment. To look more closely at the effect of plasma on cell membrane, high resolution imaging was used. We report the differences between normal human astrocytes and human glioblastoma cells by considering the membrane surface details. Our data, obtained for the first time on these cells using atomic force microscopy, argue for an architectural feature on the cell membrane, i.e. brush layers, different in normal human astrocytes as compared to glioblastoma cells. The brush layer disappears from the cell membrane surface of normal E6/E7 cells and is maintained in the glioblastoma U87 cells after plasma treatment. PMID:25803024

  8. spn-F encodes a novel protein that affects oocyte patterning and bristle morphology in Drosophila.

    PubMed

    Abdu, Uri; Bar, Dikla; Schüpbach, Trudi

    2006-04-01

    The anteroposterior and dorsoventral axes of the Drosophila embryo are established during oogenesis through the activities of Gurken (Grk), a Tgfalpha-like protein, and the Epidermal growth factor receptor (Egfr). spn-F mutant females produce ventralized eggs similar to the phenotype produced by mutations in the grk-Egfr pathway. We found that the ventralization of the eggshell in spn-F mutants is due to defects in the localization and translation of grk mRNA during mid-oogenesis. Analysis of the microtubule network revealed defects in the organization of the microtubules around the oocyte nucleus. In addition, spn-F mutants have defective bristles. We cloned spn-F and found that it encodes a novel coiled-coil protein that localizes to the minus end of microtubules in the oocyte, and this localization requires the microtubule network and a Dynein heavy chain gene. We also show that Spn-F interacts directly with the Dynein light chain Ddlc-1. Our results show that we have identified a novel protein that affects oocyte axis determination and the organization of microtubules during Drosophila oogenesis. PMID:16540510

  9. Cytokines profile and peripheral blood mononuclear cells morphology in Rett and autistic patients.

    PubMed

    Pecorelli, Alessandra; Cervellati, Franco; Belmonte, Giuseppe; Montagner, Giulia; Waldon, PhiAnh; Hayek, Joussef; Gambari, Roberto; Valacchi, Giuseppe

    2016-01-01

    A potential role for immune dysfunction in autism spectrum disorders (ASD) has been well established. However, immunological features of Rett syndrome (RTT), a genetic neurodevelopmental disorder closely related to autism, have not been well addressed yet. By using multiplex Luminex technology, a panel of 27 cytokines and chemokines was evaluated in serum from 10 RTT patients with confirmed diagnosis of MECP2 mutation (typical RTT), 12 children affected by classic autistic disorder and 8 control subjects. The cytokine/chemokine gene expression was assessed by real time PCR on mRNA of isolated peripheral blood mononuclear cells (PBMCs). Moreover, ultrastructural analysis of PBMCs was performed using transmission electron microscopy (TEM). Significantly higher serum levels of interleukin-8 (IL-8), IL-9, IL-13 were detected in RTT compared to control subjects, and IL-15 shows a trend toward the upregulation in RTT. In addition, IL-1β and VEGF were the only down-regulated cytokines in autistic patients with respect to RTT. No difference in cytokine/chemokine profile between autistic and control groups was detected. These data were also confirmed by ELISA real time PCR. At the ultrastructural level, the most severe morphological abnormalities were observed in mitochondria of both RTT and autistic PBMCs. In conclusion, our study shows a deregulated cytokine/chemokine profile together with morphologically altered immune cells in RTT. Such abnormalities were not quite as evident in autistic subjects. These findings indicate a possible role of immune dysfunction in RTT making the clinical features of this pathology related also to the immunology aspects, suggesting, therefore, novel possible therapeutic interventions for this disorder.

  10. Rosmarinic Acid and Melissa officinalis Extracts Differently Affect Glioblastoma Cells

    PubMed Central

    Ramanauskiene, Kristina; Raudonis, Raimondas

    2016-01-01

    Lemon balm (Melissa officinalis L.) has many biological effects but especially important is its neuroprotective activity. The aim of the study is to produce different extracts of Melissa officinalis and analyse their chemical composition and biological properties on rat glioblastoma C6 cells. Results revealed that rosmarinic acid (RA) is the predominant compound of lemon balm extracts. RA has cytotoxic effect on glioblastoma cells (LC50 290.5 μM after the incubation of 24 h and LC50 171.3 μM after 48 h). RA at concentration 80–130 μM suppresses the cell proliferation and has an antioxidant effect. 200 μM and higher concentrations of RA have a prooxidant effect and initiate cell death through necrosis. The aqueous extract of lemon balm is also enriched in phenolic compounds: protocatechuic, caftaric, caffeic, ferulic, and cichoric acids and flavonoid luteolin-7-glucoside. This extract at concentrations 50 μM–200 μM RA has cytotoxic activity and initiates cell death through apoptosis. Extracts prepared with 70% ethanol contain the biggest amount of active compounds. These extracts have the highest cytotoxic activity on glioblastoma cells. They initiate generation of intracellular ROS and cell death through apoptosis and necrosis. Our data suggest that differently prepared lemon balm extracts differently affect glioblastoma cells and can be used as neuroprotective agents in several therapeutic strategies.

  11. Rosmarinic Acid and Melissa officinalis Extracts Differently Affect Glioblastoma Cells

    PubMed Central

    Ramanauskiene, Kristina; Raudonis, Raimondas

    2016-01-01

    Lemon balm (Melissa officinalis L.) has many biological effects but especially important is its neuroprotective activity. The aim of the study is to produce different extracts of Melissa officinalis and analyse their chemical composition and biological properties on rat glioblastoma C6 cells. Results revealed that rosmarinic acid (RA) is the predominant compound of lemon balm extracts. RA has cytotoxic effect on glioblastoma cells (LC50 290.5 μM after the incubation of 24 h and LC50 171.3 μM after 48 h). RA at concentration 80–130 μM suppresses the cell proliferation and has an antioxidant effect. 200 μM and higher concentrations of RA have a prooxidant effect and initiate cell death through necrosis. The aqueous extract of lemon balm is also enriched in phenolic compounds: protocatechuic, caftaric, caffeic, ferulic, and cichoric acids and flavonoid luteolin-7-glucoside. This extract at concentrations 50 μM–200 μM RA has cytotoxic activity and initiates cell death through apoptosis. Extracts prepared with 70% ethanol contain the biggest amount of active compounds. These extracts have the highest cytotoxic activity on glioblastoma cells. They initiate generation of intracellular ROS and cell death through apoptosis and necrosis. Our data suggest that differently prepared lemon balm extracts differently affect glioblastoma cells and can be used as neuroprotective agents in several therapeutic strategies. PMID:27688825

  12. Hybrid Solar Cells with Prescribed Nanoscale Morphologies Based onHyperbranched Semiconductor Nanocrystals

    SciTech Connect

    Gur, Ilan; Fromer, Neil A.; Chen, Chih-Ping; Kanaras, AntoniosG.; Alivisatos, A. Paul

    2006-09-09

    In recent years, the search to develop large-area solar cells at low cost has led to research on photovoltaic (PV) systems based on nanocomposites containing conjugated polymers. These composite films can be synthesized and processed at lower costs and with greater versatility than the solid state inorganic semiconductors that comprise today's solar cells. However, the best nanocomposite solar cells are based on a complex architecture, consisting of a fine blend of interpenetrating and percolating donor and acceptor materials. Cell performance is strongly dependent on blend morphology, and solution-based fabrication techniques often result in uncontrolled and irreproducible blends, whose composite morphologies are difficult to characterize accurately. Here we incorporate 3-dimensional hyper-branched colloidal semiconductor nanocrystals in solution-processed hybrid organic-inorganic solar cells, yielding reproducible and controlled nanoscale morphology.

  13. Morphological and cytochemical determination of cell death by apoptosis

    PubMed Central

    Sobel, Burton E.; Budd, Ralph C.

    2007-01-01

    Several modes of cell death are now recognized, including necrosis, apoptosis, and autophagy. Oftentimes the distinctions between these various modes may not be apparent, although the precise mode may be physiologically important. Accordingly, it is often desirable to be able to classify the mode of cell death. Apoptosis was originally defined by structural alterations in cells observable by transmitted light and electron microscopy. Today, a wide variety of imaging and cytochemical techniques are available for the investigation of apoptosis. This review will highlight many of these methods, and provide a critique on the advantages and disadvantages associated with them for the specific identification of apoptotic cells in culture and tissues. PMID:18000678

  14. Filming a live cell by scanning electrochemical microscopy: label-free imaging of the dynamic morphology in real time

    PubMed Central

    2012-01-01

    The morphology of a live cell reflects the organization of the cytoskeleton and the healthy status of the cell. We established a label-free platform for monitoring the changing morphology of live cells in real time based on scanning electrochemical microscopy (SECM). The dynamic morphology of a live human bladder cancer cell (T24) was revealed by time-lapse SECM with dissolved oxygen in the medium solution as the redox mediator. Detailed local movements of cell membrane were presented by time-lapse cross section lines extracted from time-lapse SECM. Vivid dynamic morphology is presented by a movie made of time-lapse SECM images. The morphological change of the T24 cell by non-physiological temperature is in consistence with the morphological feature of early apoptosis. To obtain dynamic cellular morphology with other methods is difficult. The non-invasive nature of SECM combined with high resolution realized filming the movements of live cells. PMID:22436305

  15. Heterogeneity in polymer solar cells: local morphology and performance in organic photovoltaics studied with scanning probe microscopy.

    PubMed

    Groves, Chris; Reid, Obadiah G; Ginger, David S

    2010-05-18

    The use of organic photovoltaics (OPVs) could reduce production costs for solar cells because these materials are solution processable and can be manufactured by roll-to-roll printing. The nanoscale texture, or film morphology, of the donor/acceptor blends used in most OPVs is a critical variable that can dominate both the performance of new materials being optimized in the lab and efforts to move from laboratory-scale to factory-scale production. Although efficiencies of organic solar cells have improved significantly in recent years, progress in morphology optimization still occurs largely by trial and error, in part because much of our basic understanding of how nanoscale morphology affects the optoelectronic properties of these heterogeneous organic semiconductor films has to be inferred indirectly from macroscopic measurements. In this Account, we review the importance of nanoscale morphology in organic semiconductors and the use of electrical scanning probe microscopy techniques to directly probe the local optoelectronic properties of OPV devices. We have observed local heterogeneity of electronic properties and performance in a wide range of systems, including model polymer-fullerene blends such as poly(3-hexylthiophene) (P3HT) and [6,6]-phenyl-C(61)-butyric acid methyl ester (PCBM), newer polyfluorene copolymer-PCBM blends, and even all polymer donor-acceptor blends. The observed heterogeneity in local photocurrent poses important questions, chiefly what information is contained and what is lost when using average values obtained from conventional measurements on macroscopic devices and bulk samples? We show that in many cases OPVs are best thought of as a collection of nanoscopic photodiodes connected in parallel, each with their own morphological and therefore electronic and optical properties. This local heterogeneity forces us to carefully consider the adequacy of describing OPVs solely by "average" properties such as the bulk carrier mobility

  16. Substrate elasticity affects bovine satellite cell activation kinetics in vitro.

    PubMed

    Lapin, M R; Gonzalez, J M; Johnson, S E

    2013-05-01

    Satellite cells support efficient postnatal skeletal muscle hypertrophy through fusion into the adjacent muscle fiber. Nuclear contribution allows for maintenance of the fiber myonuclear domain and proficient transcription of myogenic genes. Niche growth factors affect satellite cell biology; however, the interplay between fiber elasticity and microenvironment proteins remains largely unknown. The objective of the experiment was to examine the effects of hepatocyte growth factor (HGF) and surface elasticity on bovine satellite cell (BSC) activation kinetics in vitro. Young's elastic modulus was calculated for the semimembranosus (SM) and LM muscles of young bulls (5 d; n = 8) and adult cows (27 mo; n = 4) cattle. Results indicate that LM elasticity decreased (P < 0.05) with age; no difference in Young's modulus for the SM was noted. Bovine satellite cells were seeded atop polyacrylamide bioscaffolds with surface elasticities that mimic young bull and adult cow LM or traditional cultureware. Cells were maintained in low-serum media supplemented with 5 ng/mL HGF or vehicle only for 24 or 48 h. Activation was evaluated by proliferating cell nuclear antigen (PCNA) immunocytochemistry. Results indicate that BSC maintained on rigid surfaces were activated at 24 h and refractive to HGF supplementation. By contrast, fewer (P < 0.05) BSC had exited quiescence after 24 h of culture on surfaces reflective of either young bull (8.1 ± 1.7 kPa) or adult cow (14.6 ± 1.6 kPa) LM. Supplementation with HGF promoted activation of BSC cultured on bioscaffolds as measured by an increase (P < 0.05) in PCNA immunopositive cells. Culture on pliant surfaces affected neither activation kinetics nor numbers of Paired box 7 (Pax7) immunopositive muscle stem cells (P > 0.05). However, with increasing surface elasticity, an increase (P < 0.05) in the numbers of muscle progenitors was observed. These results confirm that biophysical and biochemical signals regulate BSC activation.

  17. The G protein alpha o subunit alters morphology, growth kinetics, and phospholipid metabolism of somatic cells.

    PubMed Central

    Bloch, D B; Bonventre, J V; Neer, E J; Seidman, J G

    1989-01-01

    The physiological role of the alpha o subunit of guanine nucleotide-binding (G) protein was investigated with a murine adrenal cell line (Y1) transfected with a rat alpha o cDNA cloned in a retroviral expression vector. The parental cell line lacked detectable alpha o subunit. Expression of the alpha o cDNA in transfected cell lines was confirmed by Western blot (immunoblot) analysis. The rat alpha o subunit interacted with murine beta and gamma subunits and associated with cell membranes. Y1 cells containing large amounts of alpha o subunit had altered cellular morphology and reduced rate of cell division. In addition, GTP-gamma S-stimulated release of arachidonic acid from these cells was significantly increased compared with that in control cells. The alpha o subunit appears directly or indirectly to regulate cellular proliferation, morphology, and phospholipid metabolism. Images PMID:2511433

  18. Morphological changes in amphibian and fish cell lines infected with Andrias davidianus ranavirus.

    PubMed

    Gao, X C; Chen, Z Y; Yuan, J D; Zhang, Q Y

    2015-01-01

    Andrias davidianus ranavirus (ADRV) is an emerging viral pathogen that causes severe disease in Chinese giant salamanders, the largest extant amphibian in the world. A fish cell line, Epithelioma papulosum cyprinid (EPC), and a new amphibian cell line, Chinese giant salamander spleen cell (GSSC), were infected with ADRV and observed by light and electron microscopy. The morphological changes in these two cell lines infected with ADRV were compared. Cytopathic effect (CPE) began with rounding of the cells, progressing to cell detachment in the cell monolayer, followed by cell lysis. Significant CPE was visualized as early as 24 h post infection (hpi) in EPC cells and at 36 hpi in GSSC cells. Microscopical examination showed clear and significant CPE in EPC cells, while less extensive and irregular CPE with some adherent cells remaining was observed in GSSC cells. Following ADRV infection, CPE became more extensive. Transmission electron micrographs showed many virus particles around cytoplasmic vacuoles, formed as crystalline arrays or scattered in the cytoplasm of infected cells. Infected cells showed alteration in nuclear morphology, with condensed and marginalized nuclear chromatin on the inner aspect of the nuclear membrane and formation of a cytoplasmic viromatrix adjacent to the nucleus in both cell lines. Some virus particles were also detected in the nucleus of infected GSSC cells. Both cell lines are able to support replication of ADRV and can therefore be used to investigate amphibian ranaviruses.

  19. Dietary inulin affects the morphology but not the sodium-dependent glucose and glutamine transport in the jejunum of broilers.

    PubMed

    Rehman, H; Rosenkranz, C; Böhm, J; Zentek, J

    2007-01-01

    Inulin, a prebiotic, is a fermentable oligosaccharide that may affect the intestinal mucosal architecture and the electrophysiological parameters. The effects of a diet with added inulin were tested on the jejunal morphology and electrogenic transport of Glc and Gln from the jejunal mucosa in broilers. Short-circuit current and transmucosal tissue resistance of jejunal flaps were measured in Ussing chambers. The feeding experiment was carried out in broilers (n = 40) using 1% inulin with an application period of 5 wk. The inulin-containing diet resulted in longer jejunal villi (P < 0.05) and deeper crypts (P < 0.01) than in control birds without affecting villus:crypt depth. Basal short-circuit current value remained unaffected by dietary treatment. Inulin supplementation did not modify the electrogenic transport of Glc and Gln in the jejunal mucosa. The basal value of transmucosal tissue resistance was significantly lower (P < 0.001) in the inulin-fed group compared with the control group. In conclusion, inulin supplementation affected the jejunal mucosal architecture but did not modify the electrogenic transport of Glc and amino acid under present experimental condition.

  20. Parametric analysis of colony morphology of non-labelled live human pluripotent stem cells for cell quality control

    PubMed Central

    Kato, Ryuji; Matsumoto, Megumi; Sasaki, Hiroto; Joto, Risako; Okada, Mai; Ikeda, Yurika; Kanie, Kei; Suga, Mika; Kinehara, Masaki; Yanagihara, Kana; Liu, Yujung; Uchio-Yamada, Kozue; Fukuda, Takayuki; Kii, Hiroaki; Uozumi, Takayuki; Honda, Hiroyuki; Kiyota, Yasujiro; Furue, Miho K

    2016-01-01

    Given the difficulties inherent in maintaining human pluripotent stem cells (hPSCs) in a healthy state, hPSCs should be routinely characterized using several established standard criteria during expansion for research or therapeutic purposes. hPSC colony morphology is typically considered an important criterion, but it is not evaluated quantitatively. Thus, we designed an unbiased method to evaluate hPSC colony morphology. This method involves a combination of automated non-labelled live-cell imaging and the implementation of morphological colony analysis algorithms with multiple parameters. To validate the utility of the quantitative evaluation method, a parent cell line exhibiting typical embryonic stem cell (ESC)-like morphology and an aberrant hPSC subclone demonstrating unusual colony morphology were used as models. According to statistical colony classification based on morphological parameters, colonies containing readily discernible areas of differentiation constituted a major classification cluster and were distinguishable from typical ESC-like colonies; similar results were obtained via classification based on global gene expression profiles. Thus, the morphological features of hPSC colonies are closely associated with cellular characteristics. Our quantitative evaluation method provides a biological definition of ‘hPSC colony morphology’, permits the non-invasive monitoring of hPSC conditions and is particularly useful for detecting variations in hPSC heterogeneity. PMID:27667091

  1. Morphology control of polymer: Fullerene solar cells by nanoparticle self-assembly

    NASA Astrophysics Data System (ADS)

    Zhang, Wenluan

    achieved by having a sparse mono-layer of Fe3O4 nanoparticles (NPs) that formed a polymer depletion zone excluding P3HT away from cathode interface. Convective outflow and surface energy ordering were hypothesized to promote the NPs toward the cathode interface. By proper tuning of the NPs volume fraction added to the films the distance between two NPs can be made to be smaller than the P3HT radius of gyration to form the polymer depletion zone. PCBM molecules can then fill the space left by P3HT and help build electron transport pathways improving electron collection at cathode. The addition of NPs does not affect the PCBM agglomerate morphology, but does decrease the degree of P3HT crystallinity, so a balance between this NP effect and P3HT crystallinity has to be reached to obtain optimum device performance. To assess this NP effect in industrialized device fabrication, the rod coating method was used for a preliminary study. It is found that, the NPs do not move upward and are kinetically trapped with random a distribution within the film. In addition, the P3HT crystallinity was also reduced by adding NPs, so the device performance actually is even lower. It is hyposized that the slower drying rate in rod coating compared to spin coating traps the NPs in the film perhaps due to les convective flow. Hence, further effort is needed to realize this NP effect in large scalable device fabrication. Considering the low cost of NPs and the simple process applied to achieve this improvement, it is remarkably beneficial to organic photovoltaic industry. Further study could combine light management by using colloid particles and this NP effect to further modify morphology to obtain better solar cells. It is believed that this NP effect could be broadly applicable to other organic electronic devices like light emitting diodes, and batteries for lighting and energy storage.

  2. Prenatal exposure of diclofenac sodium affects morphology but not axon number of the median nerve of rats.

    PubMed

    Ayrancı, Ebru; Altunkaynak, Berrin Zuhal; Aktaş, Abit; Rağbetli, Murat Ç; Kaplan, Suleyman

    2013-01-01

    The present study examined the effect of DS exposure on median nerve development in rats during prenatal life. Pregnant female rats were divided into three groups: a control group, a saline group and a DS group. Offspring of these animals were divided into 2 subgroups: 4 weeks old and 20 weeks old. Nerve samples were taken from the right legs and evaluated using stereological techniques in terms of the axon number, axon cross-sectional area, and myelin thickness. No drug-dependent macroscopic abnormality was observed in the nerve. No differences were noted for axon number in the control, saline, and DS groups of the same age and gender. No gender difference was found for axon number or axon area between the other matched groups. In conclusion, prenatal exposure to diclofenac sodium does not affect axon number in rats, but can alter the morphology of the male and female median nerve. PMID:23553140

  3. Plectin isoform P1b and P1d deficiencies differentially affect mitochondrial morphology and function in skeletal muscle

    PubMed Central

    Winter, Lilli; Kuznetsov, Andrey V.; Grimm, Michael; Zeöld, Anikó; Fischer, Irmgard; Wiche, Gerhard

    2015-01-01

    Plectin, a versatile 500-kDa cytolinker protein, is essential for muscle fiber integrity and function. The most common disease caused by mutations in the human plectin gene, epidermolysis bullosa simplex with muscular dystrophy (EBS-MD), is characterized by severe skin blistering and progressive muscular dystrophy. Besides displaying pathological desmin-positive protein aggregates and degenerative changes in the myofibrillar apparatus, skeletal muscle specimens of EBS-MD patients and plectin-deficient mice are characterized by massive mitochondrial alterations. In this study, we demonstrate that structural and functional alterations of mitochondria are a primary aftermath of plectin deficiency in muscle, contributing to myofiber degeneration. We found that in skeletal muscle of conditional plectin knockout mice (MCK-Cre/cKO), mitochondrial content was reduced, and mitochondria were aggregated in sarcoplasmic and subsarcolemmal regions and were no longer associated with Z-disks. Additionally, decreased mitochondrial citrate synthase activity, respiratory function and altered adenosine diphosphate kinetics were characteristic of plectin-deficient muscles. To analyze a mechanistic link between plectin deficiency and mitochondrial alterations, we comparatively assessed mitochondrial morphology and function in whole muscle and teased muscle fibers of wild-type, MCK-Cre/cKO and plectin isoform-specific knockout mice that were lacking just one isoform (either P1b or P1d) while expressing all others. Monitoring morphological alterations of mitochondria, an isoform P1b-specific phenotype affecting the mitochondrial fusion–fission machinery and manifesting with upregulated mitochondrial fusion-associated protein mitofusin-2 could be identified. Our results show that the depletion of distinct plectin isoforms affects mitochondrial network organization and function in different ways. PMID:26019234

  4. Design of Bicontinuous Donor/Acceptor Morphologies for Use as Organic Solar Cell Active Layers

    NASA Astrophysics Data System (ADS)

    Kipp, Dylan; Mok, Jorge; Verduzco, Rafael; Ganesan, Venkat

    Two of the primary challenges limiting the marketability of organic solar cells are i) the smaller device efficiency of the organic solar cell relative to the conventional silicon-based solar cell and ii) the long term thermal instability of the device active layer. The achievement of equilibrium donor/acceptor morphologies with the characteristics believed to yield high device performance characteristics could address each of these two challenges. In this work, we present the results of a combined simulations and experiments-based approach to investigate if a conjugated BCP additive can be used to control the self-assembled morphologies taken on by conjugated polymer/PCBM mixtures. First, we use single chain in mean field Monte Carlo simulations to identify regions within the conjugated polymer/PCBM composition space in which addition of copolymers can lead to bicontinuous equilibrium morphologies with high interfacial areas and nanoscale dimensions. Second, we conduct experiments as directed by the simulations to achieve such morphologies in the PTB7 + PTB7- b-PNDI + PCBM model blend. We characterize the results of our experiments via a combination of transmission electron microscopy and X-ray scattering techniques and demonstrate that the morphologies from experiments agree with those predicted in simulations. Accordingly, these results indicate that the approach utilized represents a promising approach to intelligently design the morphologies taken on by organic solar cell active layers.

  5. Auxin regulates SNARE-dependent vacuolar morphology restricting cell size.

    PubMed

    Löfke, Christian; Dünser, Kai; Scheuring, David; Kleine-Vehn, Jürgen

    2015-03-05

    The control of cellular growth is central to multicellular patterning. In plants, the encapsulating cell wall literally binds neighbouring cells to each other and limits cellular sliding/migration. In contrast to its developmental importance, growth regulation is poorly understood in plants. Here, we reveal that the phytohormone auxin impacts on the shape of the biggest plant organelle, the vacuole. TIR1/AFBs-dependent auxin signalling posttranslationally controls the protein abundance of vacuolar SNARE components. Genetic and pharmacological interference with the auxin effect on vacuolar SNAREs interrelates with auxin-resistant vacuolar morphogenesis and cell size regulation. Vacuolar SNARE VTI11 is strictly required for auxin-reliant vacuolar morphogenesis and loss of function renders cells largely insensitive to auxin-dependent growth inhibition. Our data suggests that the adaptation of SNARE-dependent vacuolar morphogenesis allows auxin to limit cellular expansion, contributing to root organ growth rates.

  6. Dichloroacetate stimulates changes in the mitochondrial network morphology via partial mitophagy in human SH-SY5Y neuroblastoma cells.

    PubMed

    Pajuelo-Reguera, David; Alán, Lukáš; Olejár, Tomáš; Ježek, Petr

    2015-01-01

    Dichloroacetate (DCA) is beneficial in cancer therapy because it induces apoptosis and decreases cancer growth in vitro and in vivo without affecting non-cancer cells. DCA stimulates the activity of the enzyme pyruvate dehydrogenase by inhibiting pyruvate dehydrogenase kinase. Consequently, DCA promotes oxidative phosphorylation after glycolysis. Therefore, DCA produces changes in energy metabolism that could affect the mitochondrial network and mitophagy. This investigation determined the effects of DCA treatment on mitophagy in human neuroblastoma SH-SY5Y cells. SH-SY5Y cells were cultured and distributed into 3 groups: control, NH4Cl and chloroquine. Each group was treated with DCA at 0, 5, 30 and 60 mM for 16 h. Samples were analyzed for cell viability, mtDNA copy number, mitochondrial network morphology and expression of key proteins involved in mitochondrial dynamics, such as LC3b, FIS1, OPA1, PARKIN and PINK1. In all groups, DCA caused a decrease in cell viability, an induction of autophagy in a dose-dependent manner and a decrease in Tim23, FIS1 and PARKIN protein expression, leading to profound morphological changes in the mitochondrial network resulting in shorter and more fragmented filaments. However, TFAM protein levels remained unchanged. Similarly, the mitochondrial copy number was not significantly different among the treatment groups. In conclusion, DCA induces mitophagy and remodeling of the mitochondrial network. In this remodeling, DCA induces a decrease in the expression of key proteins involved in protein degradation and mitochondrial dynamics but does not significantly affect the mtDNA density. Blocking late phase autophagy increases the effects of DCA, suggesting that autophagy protects the cell, at least partially, against DCA.

  7. Factors affecting white cell content in platelet concentrates.

    PubMed

    Champion, A B; Carmen, R A

    1985-01-01

    In this study, we investigated the factors affecting white cell content in platelet concentrates. White cell yields can be reduced 50 percent by stopping platelet-rich plasma expression when the interface is 1 cm from the top of the blood bag as compared to stopping expression when the interface reaches the top of the bag. Further reductions can be achieved by careful handling during transfer of units from the centrifuge cups to expressors (after the first spin) and by carefully balancing units against each other to ensure proper rotor balance during the first spin. Following these suggestions, blood banks should be able to produce platelet concentrates with white cell yields between 2 and 6 X 10(7) and with platelet yields between 7.5 and 8 X 10(10). Transfusion of this product may reduce febrile reactions and lower the incidence of alloimmunizations. PMID:4024231

  8. Noggin 1 overexpression in retinal progenitors affects bipolar cell generation.

    PubMed

    Messina, Andrea; Bridi, Simone; Bozza, Angela; Bozzi, Yuri; Baudet, Marie-Laure; Casarosa, Simona

    2016-01-01

    Waves of Bone Morphogenetic Proteins (BMPs) and their antagonists are present during initial eye development, but their possible roles in retinogenesis are still unknown. We have recently shown that noggin 1, a BMP antagonist, renders pluripotent cells able to differentiate into retinal precursors, and might be involved in the maintenance of retinal structures in the adult vertebrate eye. Here, we report that noggin 1, differently from noggin 2 and noggin 4, is expressed during all phases of Xenopus laevis retinal development. Gain-of-function experiments by electroporation in the optic vesicle show that overexpression of noggin 1 significantly decreases the number of bipolar cells in the inner nuclear layer of the retina, without significantly affecting the generation of the other retinal cell types. Our data suggest that BMP signaling could be involved in the differentiation of retinal progenitors into specific retinal subtypes during late phases of vertebrate retinal development. PMID:27389985

  9. Concentration of Fibrin and Presence of Plasminogen Affect Proliferation, Fibrinolytic Activity, and Morphology of Human Fibroblasts and Keratinocytes in 3D Fibrin Constructs

    PubMed Central

    Reinertsen, Erik; Skinner, Michael; Wu, Benjamin

    2014-01-01

    Fibrin is a hemostatic protein found in the clotting cascade. It is used in the operating room to stop bleeding and deliver cells and growth factors to heal wounds. However, formulations of clinically approved fibrin are optimized for hemostasis, and the extent to which biochemical and physical cues in fibrin mediate skin cell behavior is not fully understood nor utilized in the design of biomaterials. To determine if the concentration of fibrinogen and the presence of plasminogen affect cell behavior relevant to wound healing, we fabricated three-dimensional fibrin constructs made from 5, 10, or 20 mg/mL of clinical fibrin or plasminogen-depleted (PD) fibrin. We cultured dermal fibroblasts or epidermal keratinocytes in these constructs. Fibroblasts proliferated similarly in both types of fibrin, but keratinocytes proliferated more in low concentrations of clinical fibrin and less in PD fibrin. Clinical fibrin constructs with fibroblasts were less stiff and degraded faster than PD fibrin constructs with fibroblasts. Similarly, keratinocytes degraded clinical fibrin, but not PD fibrin. Fibroblast spreading varied with fibrin concentration in both types of fibrin. In conclusion, the concentration of fibrinogen and the presence of plasminogen affect fibroblast and keratinocyte proliferation, morphology, and fibrin degradation. Creating materials with heterogeneous regions of fibrin formulations and concentrations could be a novel strategy for controlling the phenotype of encapsulated fibroblasts and keratinocytes, and the subsequent biomechanical properties of the construct. However, other well-investigated aspects of wound healing remain to be utilized in the design of fibrin biomaterials, such as autocrine and paracrine signaling between fibroblasts, keratinocytes, and immune cells. PMID:24738616

  10. ALDH isozymes downregulation affects cell growth, cell motility and gene expression in lung cancer cells

    PubMed Central

    Moreb, Jan S; Baker, Henry V; Chang, Lung-Ji; Amaya, Maria; Lopez, M Cecilia; Ostmark, Blanca; Chou, Wayne

    2008-01-01

    Background Aldehyde dehydrogenase isozymes ALDH1A1 and ALDH3A1 are highly expressed in non small cell lung cancer. Neither the mechanisms nor the biologic significance for such over expression have been studied. Methods We have employed oligonucleotide microarrays to analyze changes in gene profiles in A549 lung cancer cell line in which ALDH activity was reduced by up to 95% using lentiviral mediated expression of siRNA against both isozymes (Lenti 1+3). Stringent analysis methods were used to identify gene expression patterns that are specific to the knock down of ALDH activity and significantly different in comparison to wild type A549 cells (WT) or cells similarly transduced with green fluorescent protein (GFP) siRNA. Results We confirmed significant and specific down regulation of ALDH1A1 and ALDH3A1 in Lenti 1+3 cells and in comparison to 12 other ALDH genes detected. The results of the microarray analysis were validated by real time RT-PCR on RNA obtained from Lenti 1+3 or WT cells treated with ALDH activity inhibitors. Detailed functional analysis was performed on 101 genes that were significantly different (P < 0.001) and their expression changed by ≥ 2 folds in the Lenti 1+3 group versus the control groups. There were 75 down regulated and 26 up regulated genes. Protein binding, organ development, signal transduction, transcription, lipid metabolism, and cell migration and adhesion were among the most affected pathways. Conclusion These molecular effects of the ALDH knock-down are associated with in vitro functional changes in the proliferation and motility of these cells and demonstrate the significance of ALDH enzymes in cell homeostasis with a potentially significant impact on the treatment of lung cancer. PMID:19025616

  11. Culture materials affect ex vivo expansion of hematopoietic progenitor cells.

    PubMed

    LaIuppa, J A; McAdams, T A; Papoutsakis, E T; Miller, W M

    1997-09-01

    Ex vivo expansion of hematopoietic cells is important for applications such as cancer treatment, gene therapy, and transfusion medicine. While cell culture systems are widely used to evaluate the biocompatibility of materials for implantation, the ability of materials to support proliferation of primary human cells in cultures for reinfusion into patients has not been addressed. We screened a variety of commercially available polymer (15 types), metal (four types), and glass substrates for their ability to support expansion of hematopoietic cells when cultured under conditions that would be encountered in a clinical setting. Cultures of peripheral blood (PB) CD34+ cells and mononuclear cells (MNC) were evaluated for expansion of total cells and colony-forming unit-granulocyte monocyte (CFU-GM; progenitors committed to the granulocyte and/or monocyte lineage). Human hematopoietic cultures in serum-free medium were found to be extremely sensitive to the substrate material. The only materials tested that supported expansion at or near the levels of polystyrene were tissue culture polystyrene, Teflon perfluoroalkoxy, Teflon fluorinated ethylene propylene, cellulose acetate, titanium, new polycarbonate, and new polymethylpentene. MNC were less sensitive to the substrate materials than the primitive CD34+ progenitors, although similar trends were seen for expansion of the two cell populations on the substrates tested. CFU-GM expansion was more sensitive to substrate materials than was total cell expansion. The detrimental effects of a number of the materials on hematopoietic cultures appear to be caused by protein adsorption and/or leaching of toxins. Factors such as cleaning, sterilization, and reuse significantly affected the performance of some materials as culture substrates. We also used PB CD34+ cell cultures to examine the biocompatibility of gas-permeable cell culture and blood storage bags and several types of tubing commonly used with biomedical equipment

  12. Clear Cell Renal Cell Carcinoma With Borderline Features of Clear Cell Papillary Renal Cell Carcinoma: Combined Morphologic, Immunohistochemical, and Cytogenetic Analysis.

    PubMed

    Williamson, Sean R; Gupta, Nilesh S; Eble, John N; Rogers, Craig G; Michalowski, Susan; Zhang, Shaobo; Wang, Mingsheng; Grignon, David J; Cheng, Liang

    2015-11-01

    Clear cell papillary renal cell carcinoma is increasingly recognized as a distinct tumor with unique morphology, immunohistochemistry, and cytogenetics. Histopathology often mimics clear cell renal cell carcinoma; however, metastasis has not been reported, emphasizing the clinical value of recognizing these likely nonaggressive tumors. We studied tumors with borderline morphology of clear cell papillary renal cell carcinoma, utilizing immunohistochemistry and fluorescence in situ hybridization or karyotyping. Tumors from 22 patients (ages 33 to 82 y) were analyzed. Clear cell papillary renal cell carcinoma-like morphology varied from 10% to 90% of the tumor (median 25%). Sources of resemblance included: branched glands (95%), nuclear alignment (68%), small papillary tufts (32%), focal branching papillae (27%), and prominent papillary structures (9%). Carbonic anhydrase IX uniformly revealed diffuse positivity. Staining for cytokeratin 7 (CK7) was focal (64%) or negative (18%) in most tumors (82%); however, >50% labeling was present in 4 (18%). Reactivity for both CD10 and α-methyl-acyl-CoA-racemase (AMACR) was usually present (median 80% and 60% of cells). Seven tumors showed reactivity for high-molecular weight keratin (32%). Chromosome 3p loss was confirmed in 15 tumors (68%), including 4/7 with labeling for high-molecular weight keratin or >50% reactivity for CK7. A discordant immunohistochemical pattern typically correlates with loss of material from chromosome 3p in tumors with incomplete morphology of clear cell papillary renal cell carcinoma, supporting classification as clear cell renal cell carcinoma. Diffuse labeling for CK7 can uncommonly be observed in clear cell renal cell carcinomas confirmed to have chromosome 3p loss, although these do not exhibit the expected staining pattern of clear cell papillary renal cell carcinoma, including positivity for CD10 and AMACR. PMID:26457355

  13. Mps1 (Monopolar Spindle 1) Protein Inhibition Affects Cellular Growth and Pro-Embryogenic Masses Morphology in Embryogenic Cultures of Araucaria angustifolia (Araucariaceae).

    PubMed

    Douétts-Peres, Jackellinne C; Cruz, Marco Antônio L; Reis, Ricardo S; Heringer, Angelo S; de Oliveira, Eduardo A G; Elbl, Paula M; Floh, Eny I S; Silveira, Vanildo; Santa-Catarina, Claudete

    2016-01-01

    Somatic embryogenesis has been shown to be an efficient tool for studying processes based on cell growth and development. The fine regulation of the cell cycle is essential for proper embryo formation during the process of somatic embryogenesis. The aims of the present work were to identify and perform a structural and functional characterization of Mps1 and to analyze the effects of the inhibition of this protein on cellular growth and pro-embryogenic mass (PEM) morphology in embryogenic cultures of A. angustifolia. A single-copy Mps1 gene named AaMps1 was retrieved from the A. angustifolia transcriptome database, and through a mass spectrometry approach, AaMps1 was identified and quantified in embryogenic cultures. The Mps1 inhibitor SP600125 (10 μM) inhibited cellular growth and changed PEMs, and these effects were accompanied by a reduction in AaMps1 protein levels in embryogenic cultures. Our work has identified the Mps1 protein in a gymnosperm species for the first time, and we have shown that inhibiting Mps1 affects cellular growth and PEM differentiation during A. angustifolia somatic embryogenesis. These data will be useful for better understanding cell cycle control during somatic embryogenesis in plants. PMID:27064899

  14. Mps1 (Monopolar Spindle 1) Protein Inhibition Affects Cellular Growth and Pro-Embryogenic Masses Morphology in Embryogenic Cultures of Araucaria angustifolia (Araucariaceae)

    PubMed Central

    Douétts-Peres, Jackellinne C.; Cruz, Marco Antônio L.; Reis, Ricardo S.; Heringer, Angelo S.; de Oliveira, Eduardo A. G.; Elbl, Paula M.; Floh, Eny I. S.; Silveira, Vanildo

    2016-01-01

    Somatic embryogenesis has been shown to be an efficient tool for studying processes based on cell growth and development. The fine regulation of the cell cycle is essential for proper embryo formation during the process of somatic embryogenesis. The aims of the present work were to identify and perform a structural and functional characterization of Mps1 and to analyze the effects of the inhibition of this protein on cellular growth and pro-embryogenic mass (PEM) morphology in embryogenic cultures of A. angustifolia. A single-copy Mps1 gene named AaMps1 was retrieved from the A. angustifolia transcriptome database, and through a mass spectrometry approach, AaMps1 was identified and quantified in embryogenic cultures. The Mps1 inhibitor SP600125 (10 μM) inhibited cellular growth and changed PEMs, and these effects were accompanied by a reduction in AaMps1 protein levels in embryogenic cultures. Our work has identified the Mps1 protein in a gymnosperm species for the first time, and we have shown that inhibiting Mps1 affects cellular growth and PEM differentiation during A. angustifolia somatic embryogenesis. These data will be useful for better understanding cell cycle control during somatic embryogenesis in plants. PMID:27064899

  15. Mps1 (Monopolar Spindle 1) Protein Inhibition Affects Cellular Growth and Pro-Embryogenic Masses Morphology in Embryogenic Cultures of Araucaria angustifolia (Araucariaceae).

    PubMed

    Douétts-Peres, Jackellinne C; Cruz, Marco Antônio L; Reis, Ricardo S; Heringer, Angelo S; de Oliveira, Eduardo A G; Elbl, Paula M; Floh, Eny I S; Silveira, Vanildo; Santa-Catarina, Claudete

    2016-01-01

    Somatic embryogenesis has been shown to be an efficient tool for studying processes based on cell growth and development. The fine regulation of the cell cycle is essential for proper embryo formation during the process of somatic embryogenesis. The aims of the present work were to identify and perform a structural and functional characterization of Mps1 and to analyze the effects of the inhibition of this protein on cellular growth and pro-embryogenic mass (PEM) morphology in embryogenic cultures of A. angustifolia. A single-copy Mps1 gene named AaMps1 was retrieved from the A. angustifolia transcriptome database, and through a mass spectrometry approach, AaMps1 was identified and quantified in embryogenic cultures. The Mps1 inhibitor SP600125 (10 μM) inhibited cellular growth and changed PEMs, and these effects were accompanied by a reduction in AaMps1 protein levels in embryogenic cultures. Our work has identified the Mps1 protein in a gymnosperm species for the first time, and we have shown that inhibiting Mps1 affects cellular growth and PEM differentiation during A. angustifolia somatic embryogenesis. These data will be useful for better understanding cell cycle control during somatic embryogenesis in plants.

  16. The Absence of Pupylation (Prokaryotic Ubiquitin-Like Protein Modification) Affects Morphological and Physiological Differentiation in Streptomyces coelicolor

    PubMed Central

    Seghezzi, Nicolas; Duchateau, Magalie; Gominet, Myriam; Kofroňová, Olga; Benada, Oldřich; Mazodier, Philippe

    2015-01-01

    ABSTRACT Protein turnover is essential in all living organisms for the maintenance of normal cell physiology. In eukaryotes, most cellular protein turnover involves the ubiquitin-proteasome pathway, in which proteins tagged with ubiquitin are targeted to the proteasome for degradation. In contrast, most bacteria lack a proteasome but harbor proteases for protein turnover. However, some actinobacteria, such as mycobacteria, possess a proteasome in addition to these proteases. A prokaryotic ubiquitination-like tagging process in mycobacteria was described and was named pupylation: proteins are tagged with Pup (prokaryotic ubiquitin-like protein) and directed to the proteasome for degradation. We report pupylation in another actinobacterium, Streptomyces coelicolor. Both the morphology and life cycle of Streptomyces species are complex (formation of a substrate and aerial mycelium followed by sporulation), and these bacteria are prolific producers of secondary metabolites with important medicinal and agricultural applications. The genes encoding the pupylation system in S. coelicolor are expressed at various stages of development. We demonstrated that pupylation targets numerous proteins and identified 20 of them. Furthermore, we established that abolition of pupylation has substantial effects on morphological and metabolic differentiation and on resistance to oxidative stress. In contrast, in most cases, a proteasome-deficient mutant showed only modest perturbations under the same conditions. Thus, the phenotype of the pup mutant does not appear to be due solely to defective proteasomal degradation. Presumably, pupylation has roles in addition to directing proteins to the proteasome. IMPORTANCE Streptomyces spp. are filamentous and sporulating actinobacteria, remarkable for their morphological and metabolic differentiation. They produce numerous bioactive compounds, including antifungal, antibiotic, and antitumor compounds. There is therefore considerable interest in

  17. Morphological evidence of neutrophil-tumor cell phagocytosis (cannibalism) in human gastric adenocarcinomas.

    PubMed

    Caruso, R A; Muda, A O; Bersiga, A; Rigoli, L; Inferrera, C

    2002-01-01

    The phenomenon of neutrophil-tumor cell emperipolesis or phagocytosis has been documented by light microscopy in various human carcinomas, but little is known about the cellular pathological processes and the morphological changes involved. In an attempt to clarify the nature of this phenomenon, the authors' ultrastructural studies on the relationships among neutrophils and tumor cells in human gastric carcinomas are reviewed and analyzed. At the electron microscopy level, apoptotic neutrophils were found within vacuoles of adenocarcinoma cells in 2 cases. They showed either early apoptotic morphology with perinuclear chromatin aggregation but cytoplasm integrity or late apoptotic morphology with uniform, collapsed nucleus and tightly packed cytoplasmic granules. A light microscopy review of 200 cases of resected gastric carcinomas identified 22 cases (11%) that were characterized by neutrophil-tumor cell phagocytosis (cannibalism). TUNEL staining confirmed the presence of apoptotic neutrophils within the cytoplasm of the tumor cells. This study provides light and electron microscopic evidence of apoptotic neutrophils phagocytosed by gastric adenocarcinoma cells. The morphological features of neutrophil-tumor cell phagocytosis (cannibalism) would suggest a particular mechanism of tumor-immune escape in human gastric carcinoma. PMID:12396242

  18. Morphological adaptation and inhibition of cell division during stationary phase in Caulobacter crescentus.

    PubMed

    Wortinger, M A; Quardokus, E M; Brun, Y V

    1998-08-01

    During exponential growth, each cell cycle of the alpha-purple bacterium Caulobacter crescentus gives rise to two different cell types: a motile swarmer cell and a sessile stalked cell. When cultures of C. crescentus are grown for extended periods in complex (PYE) medium, cells undergo dramatic morphological changes and display increased resistance to stress. After cultures enter stationary phase, most cells are arrested at the predivisional stage. For the first 6-8 days after inoculation, the colony-forming units (cfu) steadily decrease from 10(9) cfu ml(-1) to a minimum of 3x10(7) cfu ml(-1) after which cells gradually adopt an elongated helical morphology. For days 9-12, the cfu of the culture increase and stabilize around 2 x 10(8) cfu ml(-1). The viable cells have an elongated helical morphology with no constrictions and an average length of 20 microm, which is 15-20 times longer than exponentially growing cells. The level of the cell division initiation protein FtsZ decreases during the first week in stationary phase and remains at a low constant level consistent with the lack of cell division. When resuspended in fresh medium, the elongated cells return to normal size and morphology within 12 h. Cells that have returned from stationary phase proceed through the same developmental changes when they are again grown for an extended period and have not acquired a heritable growth advantage in stationary phase (GASP) compared with overnight cultures. We conclude that the changes observed in prolonged cultures are the result of entry into a new developmental pathway and are not due to mutation. PMID:9767565

  19. Understanding Polymer-Fullerene Morphology in Organic Solar Cells via Photoluminescence, Raman Scattering, and Spectroscopic Imaging

    NASA Astrophysics Data System (ADS)

    Carach, Christopher Andrew

    Understanding and controlling carrier transport in conjugated polymer films and composites is critical to the development and application of plastic solar cells. Recent efforts have focused on "bulk heterojunction" structures where a conjugated polymer donor is mixed at the nanoscale with a fullerene acceptor to achieve large interfacial areas for exciton splitting. In these systems, fabrication protocols dramatically affect device efficiency and charge transport is intimately tied to film morphology through local order, domain formation, and compositional heterogeneity. We employ both far-field and confocal/near-field optical spectroscopy (absorbance, low-temperature photoluminescence, Raman) to study chain order (aggregation, pi-stacking), photo-oxidation, and local morphology in conjugated polymer (PPV and polythiophene) -- fullerene (PCBM) blends. Through quantitative analysis of exciton bandwidths, emission intensity, and vibronic lineshapes, we demonstrate that competition exists between the chemical "disordering" effect of photo-degradation and the physical "ordering" effect of aggregation, each of which dominate under different processing conditions. Large changes in photoluminescence and Raman show that PCBM begins to significantly hinder intra-chain planarization and inter-chain pi-overlap at a critical PCBM weight fraction. Furthermore, the critical weight fraction is a function of the polymer regiochemistry, occurring at lower PCBM weight fractions for a more regio-random polymer. Mild thermal annealing of blended films was seen to restore order, which results from PCBM phase segregation (lower dispersion) and growth of polymer aggregates. Spatially resolved spectral analysis of photoluminescence was also used to map fullerene diffusion and agglomeration as well as detect local changes in interfacial contact between donor and acceptor domains due to thermal annealing.

  20. Morphological characteristics of cultured fresh and thawed pericardium cells.

    PubMed

    Maslova, Olga; Fedevych, Oleg; Shuvalova, Nadiia; Deryabina, Olena; Zhovnir, Volodymyr; Novak, Miroslav; Kruzliak, Peter

    2016-06-01

    The need for selection of the optimal material for the manufacturing of cardio-patches can be resolved by the use of cryostored autologous pericardial tissue. This short communication is a concise fragment of a large-scale research and demonstrates only the efficiency of cell culturing before and after pericardial preservation in the low temperature conditions.

  1. Cell morphology classification in phase contrast microscopy image reducing halo artifact

    NASA Astrophysics Data System (ADS)

    Kang, Mi-Sun; Song, Soo-Min; Lee, Hana; Kim, Myoung-Hee

    2012-03-01

    Since the morphology of tumor cells is a good indicator of their invasiveness, we used time-lapse phase-contrast microscopy to examine the morphology of tumor cells. This technique enables long-term observation of the activity of live cells without photobleaching and phototoxicity which is common in other fluorescence-labeled microscopy. However, it does have certain drawbacks in terms of imaging. Therefore, we first corrected for non-uniform illumination artifacts and then we use intensity distribution information to detect cell boundary. In phase contrast microscopy image, cell is normally appeared as dark region surrounded by bright halo ring. Due to halo artifact is minimal around the cell body and has non-symmetric diffusion pattern, we calculate cross sectional plane which intersects center of each cell and orthogonal to first principal axis. Then, we extract dark cell region by analyzing intensity profile curve considering local bright peak as halo area. Finally, we examined cell morphology to classify tumor cells as malignant and benign.

  2. Investigation of cell morphology for disease diagnostics via high content screening

    NASA Astrophysics Data System (ADS)

    Khatau, Shyam

    2013-03-01

    Ninety percent of all cancer-related deaths are caused by metastatic disease, i.e. the spreading of a subset of cells from a primary tumor in an organ to distal sites in other organs. Understanding this progression from localized to metastatic disease is essential for further developing effective therapeutic and treatment strategies. However, despite research efforts, no distinct genetic, epigenetic, or proteomic signature of cancer metastasis has been identified so far. Metastasis is a physical event: through invasion and migration through the dense, tortuous stromal matrix, intravasation, shear forces of blood flow, successful re-attachment to blood vessel walls, migration, the colonization of a distal site, and, finally, reactivation following dormancy, metastatic cells may share precise physical properties. Cell morphology is the most direct physical property that can be measured. In this work, we develop a high throughput cell phenotyping process and investigate the morphological signature of primary tumor cells and liver metastatic pancreatic cancer cells.

  3. Puromycin insensitive leucyl-specific aminopeptidase (PILSAP) affects RhoA activation in endothelial cells.

    PubMed

    Suzuki, Takahiro; Abe, Mayumi; Miyashita, Hiroki; Kobayashi, Toshimitsu; Sato, Yasufumi

    2007-06-01

    Puromycin insensitive leucyl-specific aminopeptidase (PILSAP) expressed in endothelial cells (ECs) plays an important role in angiogenesis due to its involvement in migration, proliferation and network formation. Here we examined the biological function of PILSAP with respect to EC morphogenesis and the related intracellular signaling for this process. When mouse endothelial MSS31 cells were cultured, a dominant negative PILSAP mutant converted cell shape to disk-like morphology, blocked stress fiber formation, and augmented membrane ruffling in random directions. These phenotypic changes led us to test whether PILSAP affected activities of Rho family small G-proteins. Abrogation of PILSAP enzymatic activity or its expression attenuated RhoA but not Rac1 activation during cell adhesion. This attenuation of RhoA activation was also evident when G-protein coupled receptors such as proteinase-activated receptor or lysophosphatidic acid receptor were activated in ECs. These results indicate that PILSAP affects RhoA activation and that influences the proper function of ECs.

  4. Melanopsin, photosensitive ganglion cells, and seasonal affective disorder.

    PubMed

    Roecklein, Kathryn A; Wong, Patricia M; Miller, Megan A; Donofry, Shannon D; Kamarck, Marissa L; Brainard, George C

    2013-03-01

    In two recent reports, melanopsin gene variations were associated with seasonal affective disorder (SAD), and in changes in the timing of sleep and activity in healthy individuals. New studies have deepened our understanding of the retinohypothalamic tract, which translates environmental light received by the retina into neural signals sent to a set of nonvisual nuclei in the brain that are responsible for functions other than sight including circadian, neuroendocrine and neurobehavioral regulation. Because this pathway mediates seasonal changes in physiology, behavior, and mood, individual variations in the pathway may explain why approximately 1-2% of the North American population develops mood disorders with a seasonal pattern (i.e., Major Depressive and Bipolar Disorders with a seasonal pattern, also known as seasonal affective disorder/SAD). Components of depression including mood changes, sleep patterns, appetite, and cognitive performance can be affected by the biological and behavioral responses to light. Specifically, variations in the gene sequence for the retinal photopigment, melanopsin, may be responsible for significant increased risk for mood disorders with a seasonal pattern, and may do so by leading to changes in activity and sleep timing in winter. The retinal sensitivity of SAD is hypothesized to be decreased compared to controls, and that further decrements in winter light levels may combine to trigger depression in winter. Here we outline steps for new research to address the possible role of melanopsin in seasonal affective disorder including chromatic pupillometry designed to measure the sensitivity of melanopsin containing retinal ganglion cells.

  5. Induction of morphological changes in death-induced cancer cells monitored by holographic microscopy.

    PubMed

    El-Schich, Zahra; Mölder, Anna; Tassidis, Helena; Härkönen, Pirkko; Falck Miniotis, Maria; Gjörloff Wingren, Anette

    2015-03-01

    We are using the label-free technique of holographic microscopy to analyze cellular parameters including cell number, confluence, cellular volume and area directly in the cell culture environment. We show that death-induced cells can be distinguished from untreated counterparts by the use of holographic microscopy, and we demonstrate its capability for cell death assessment. Morphological analysis of two representative cell lines (L929 and DU145) was performed in the culture flasks without any prior cell detachment. The two cell lines were treated with the anti-tumour agent etoposide for 1-3days. Measurements by holographic microscopy showed significant differences in average cell number, confluence, volume and area when comparing etoposide-treated with untreated cells. The cell volume of the treated cell lines was initially increased at early time-points. By time, cells decreased in volume, especially when treated with high doses of etoposide. In conclusion, we have shown that holographic microscopy allows label-free and completely non-invasive morphological measurements of cell growth, viability and death. Future applications could include real-time monitoring of these holographic microscopy parameters in cells in response to clinically relevant compounds.

  6. Monocytoid leukemia cell line CTV-1: morphological, immunological and isoenzymatic characteristics.

    PubMed

    Drexler, H G; Gaedicke, G; Maeda, S; Chen, P M; Minowada, J

    1986-01-01

    The human leukemia cell line CTV-1 was established from a case of acute monoblastic leukemia (AMoL). We analyzed the phenotypic marker profile of the CTV-1 cells in their original, untreated state and during induction of differentiation with the phorbolester 12-0-tetradecanoylphorbol 13-acetate (TPA). TPA led to morphological changes with signs of differentiation. Cell proliferation decreased in a dose-dependent fashion during exposure to TPA. In the surface marker analysis using a panel of 45 monoclonal antibodies (MoAbs) and several polyclonal antisera, CTV-1 cells were negative for markers of the T- and B-cell lineages, and were positive for several, but not all, myelomonocytic markers. Although the cells were reactive with the MoAb Leu-7 which identifies natural killer (NK) T-cells, no NK activity was detected. In the isoenzyme analysis of the four enzymes carboxylic esterase, acid phosphatase, hexosaminidase and lactate dehydrogenase (LDH) performed by isoelectric focusing on polyacrylamide gels, CTV-1 cells displayed isoenzyme profiles of immature myeloid cells. The overall marker profile of CTV-1 cells demonstrated cells of monocytoid origin arrested at a very early stage of differentiation, possibly close to the stage of precursor cells. As compared to other myelomonocytic cell lines, CTV-1 cells showed unusual morphological, immunological, functional and biochemical features and appeared to be relatively insensitive to treatment with TPA, although some alterations of the phenotype could be induced. PMID:3458274

  7. Morphologic characteristics of processes of nucleus pulposus cells in adult human intervertebral disc

    NASA Astrophysics Data System (ADS)

    Liu, Xiaoyun; Wu, Xinghuo; Hui, Liu; Xu, Weihua; Liu, Xianze; Yang, Shuhua

    2008-12-01

    To explore morphologic characterizatics of cellular processes from adult human nucleus pulposus cells, the nucleus pulposus of adult human intervertebral disc were obtained from 8 patients (Thompson's grade I~II) and then the tissues specimens were carried out by frozen section and electron microscopic section as well as cell isolation and cultured, processes of nucleus pulposus cells were examined using light microscopy, laser scanning confocal microscopy and transmission electron microscopy. When examined at both the confocal and electron microscope level, all the cells possessed the processes and adjacent nucleus pulposus cells processes possessed a gap junction. But elongated and round cells can be examined when NP cells were monolayer cultured. The rate of elongated cells to round cells is 2.3 to 1. The elongated cells protrude along with the long axis of cell body without second processes. Dendritic processes of round cells protrude to all directions from the cell body with multiple-level processes. Processes are one of the morphologic characteristics of intervertebral disc cells which are different from articular cartilage chondrocytes. The research on processes functions will be helpful to understand pathomechanism of intervertebral disc degradation and open a new approach for cytobiology treatment of the intervertebral disc diseases.

  8. Surface chemical functionalities affect the behavior of human adipose-derived stem cells in vitro

    NASA Astrophysics Data System (ADS)

    Liu, Xujie; Feng, Qingling; Bachhuka, Akash; Vasilev, Krasimir

    2013-04-01

    This study examines the effect of surface chemical functionalities on the behavior of human adipose-derived stem cells (hASCs) in vitro. Plasma polymerized films rich in amine (sbnd NH2), carboxyl (sbnd COOH) and methyl (sbnd CH3), were generated on hydroxyapatite (HAp) substrates. The surface chemical functionalities were characterized by X-ray photoelectron spectroscopy (XPS). The ability of different substrates to absorb proteins was evaluated. The results showed that substrates modified with hydrophilic functional group (sbnd COOH and sbnd NH2) can absorb more proteins than these modified with more hydrophobic functional group (sbnd CH3). The behavior of human adipose-derived stem cells (hASCs) cultured on different substrates was investigated in vitro: cell counting kit-8 (CCK-8) analysis was used to characterize cell proliferation, scanning electronic microscopy (SEM) analysis was used to characterize cell morphology and alkaline phosphatase (ALP) activity analysis was used to account for differentiation. The results of this study demonstrated that the sbnd NH2 modified surfaces encourage osteogenic differentiation; the sbnd COOH modified surfaces promote cell adhesion and spreading and the sbnd CH3 modified surfaces have the lowest ability to induce osteogenic differentiation. These findings confirmed that the surface chemical states of biomaterials can affect the behavior of hASCs in vitro.

  9. Differential regulation of morphology and stemness of mouse embryonic stem cells by substrate stiffness and topography.

    PubMed

    Lü, Dongyuan; Luo, Chunhua; Zhang, Chen; Li, Zhan; Long, Mian

    2014-04-01

    The maintenance of stem cell pluripotency or stemness is crucial to embryonic development and differentiation. The mechanical or physical microenvironment of stem cells, which includes extracellular matrix stiffness and topography, regulates cell morphology and stemness. Although a growing body of evidence has shown the importance of these factors in stem cell differentiation, the impact of these biophysical or biomechanical regulators remains insufficiently characterized. In the present study, we applied a micro-fabricated polyacrylamide hydrogel substrate with two elasticities and three topographies to systematically test the morphology, proliferation, and stemness of mESCs. The independent or combined impact of the two factors on specific cell functions was analyzed. Cells are able to grow effectively on both polystyrene and polyacrylamide substrates in the absence of feeder cells. Substrate stiffness is predominant in preserving stemness by enhancing Oct-4 and Nanog expression on a soft polyacrylamide substrate. Topography is also a critical factor for manipulating stemness via the formation of a relatively flattened colony on a groove or pillar substrate and a spheroid colony on a hexagonal substrate. Although topography is less effective on soft substrates, it plays a role in retaining cell stemness on stiff, hexagonal or pillar-shaped substrates. mESCs also form, in a timely manner, a 3D structure on groove or hexagonal substrates. These results further the understanding of stem cell morphology and stemness in a microenvironment that mimics physiological conditions.

  10. New paradigm to assess brain cell morphology by diffusion-weighted MR spectroscopy in vivo

    PubMed Central

    Palombo, Marco; Ligneul, Clémence; Najac, Chloé; Le Douce, Juliette; Flament, Julien; Escartin, Carole; Hantraye, Philippe; Brouillet, Emmanuel; Bonvento, Gilles; Valette, Julien

    2016-01-01

    The brain is one of the most complex organs, and tools are lacking to assess its cellular morphology in vivo. Here we combine original diffusion-weighted magnetic resonance (MR) spectroscopy acquisition and novel modeling strategies to explore the possibility of quantifying brain cell morphology noninvasively. First, the diffusion of cell-specific metabolites is measured at ultra-long diffusion times in the rodent and primate brain in vivo to observe how cell long-range morphology constrains metabolite diffusion. Massive simulations of particles diffusing in synthetic cells parameterized by morphometric statistics are then iterated to fit experimental data. This method yields synthetic cells (tentatively neurons and astrocytes) that exhibit striking qualitative and quantitative similarities with histology (e.g., using Sholl analysis). With our approach, we measure major interspecies difference regarding astrocytes, whereas dendritic organization appears better conserved throughout species. This work suggests that the time dependence of metabolite diffusion coefficient allows distinguishing and quantitatively characterizing brain cell morphologies noninvasively. PMID:27226303

  11. Imaging Nuclear Morphology and Organization in Cleared Plant Tissues Treated with Cell Cycle Inhibitors.

    PubMed

    de Souza Junior, José Dijair Antonino; de Sa, Maria Fatima Grossi; Engler, Gilbert; Engler, Janice de Almeida

    2016-01-01

    Synchronization of root cells through chemical treatment can generate a large number of cells blocked in specific cell cycle phases. In plants, this approach can be employed for cell suspension cultures and plant seedlings. To identify plant cells in the course of the cell cycle, especially during mitosis in meristematic tissues, chemical inhibitors can be used to block cell cycle progression. Herein, we present a simplified and easy-to-apply protocol to visualize mitotic figures, nuclei morphology, and organization in whole Arabidopsis root apexes. The procedure is based on tissue clearing, and fluorescent staining of nuclear DNA with DAPI. The protocol allows carrying out bulk analysis of nuclei and cell cycle phases in root cells and will be valuable to investigate mutants like overexpressing lines of genes disturbing the plant cell cycle.

  12. Photoinduced cell morphology alterations quantified within adipose tissues by spectral optical coherence tomography.

    PubMed

    Yanina, Irina Yu; Trunina, Natalia A; Tuchin, Valery V

    2013-11-01

    Morphological changes of the adipose tissue at phototreatment are studied in vitro using optical coherence tomography. The 200 to 600 μm fat tissue slices are used in the experiments. The observed change in the tissue structure was associated with fat cell lipolysis and destruction caused by the photodynamic effect. It is found that overall heating of a sample from room to physiological temperature leads to deeper and faster morphology tissue changes if other processing conditions are kept constant. These data support the hypothesis that photodynamic/photothermal treatment induces fat cell lipolysis during some period after treatment.

  13. Nanomechanical clues from morphologically normal cervical squamous cells could improve cervical cancer screening

    NASA Astrophysics Data System (ADS)

    Geng, Li; Feng, Jiantao; Sun, Quanmei; Liu, Jing; Hua, Wenda; Li, Jing; Ao, Zhuo; You, Ke; Guo, Yanli; Liao, Fulong; Zhang, Youyi; Guo, Hongyan; Han, Jinsong; Xiong, Guangwu; Zhang, Lufang; Han, Dong

    2015-09-01

    Applying an atomic force microscope, we performed a nanomechanical analysis of morphologically normal cervical squamous cells (MNSCs) which are commonly used in cervical screening. Results showed that nanomechanical parameters of MNSCs correlate well with cervical malignancy, and may have potential in cancer screening to provide early diagnosis.Applying an atomic force microscope, we performed a nanomechanical analysis of morphologically normal cervical squamous cells (MNSCs) which are commonly used in cervical screening. Results showed that nanomechanical parameters of MNSCs correlate well with cervical malignancy, and may have potential in cancer screening to provide early diagnosis. Electronic supplementary information (ESI) available. See DOI: 10.1039/c5nr03662c

  14. Genome rearrangement affects RNA virus adaptability on prostate cancer cells

    PubMed Central

    Pesko, Kendra; Voigt, Emily A.; Swick, Adam; Morley, Valerie J.; Timm, Collin; Yin, John; Turner, Paul E.

    2015-01-01

    Gene order is often highly conserved within taxonomic groups, such that organisms with rearranged genomes tend to be less fit than wild type gene orders, and suggesting natural selection favors genome architectures that maximize fitness. But it is unclear whether rearranged genomes hinder adaptability: capacity to evolutionarily improve in a new environment. Negative-sense non-segmented RNA viruses (order Mononegavirales) have specific genome architecture: 3′ UTR – core protein genes – envelope protein genes – RNA-dependent RNA-polymerase gene – 5′ UTR. To test how genome architecture affects RNA virus evolution, we examined vesicular stomatitis virus (VSV) variants with the nucleocapsid (N) gene moved sequentially downstream in the genome. Because RNA polymerase stuttering in VSV replication causes greater mRNA production in upstream genes, N gene translocation toward the 5′ end leads to stepwise decreases in N transcription, viral replication and progeny production, and also impacts the activation of type 1 interferon mediated antiviral responses. We evolved VSV gene-order variants in two prostate cancer cell lines: LNCap cells deficient in innate immune response to viral infection, and PC-3 cells that mount an IFN stimulated anti-viral response to infection. We observed that gene order affects phenotypic adaptability (reproductive growth; viral suppression of immune function), especially on PC-3 cells that strongly select against virus infection. Overall, populations derived from the least-fit ancestor (most-altered N position architecture) adapted fastest, consistent with theory predicting populations with low initial fitness should improve faster in evolutionary time. Also, we observed correlated responses to selection, where viruses improved across both hosts, rather than suffer fitness trade-offs on unselected hosts. Whole genomics revealed multiple mutations in evolved variants, some of which were conserved across selective environments for a

  15. Morphological characterization of cells in concentrated suspensions using multispectral diffuse optical tomography

    PubMed Central

    Hajihashemi, Mohammad Reza; Li, Xiaoqi; Jiang, Huabei

    2013-01-01

    Based on a non-spherical model of particle scattering, we investigate the capabilities and limitations of a T-matrix based inverse algorithm to morphologically characterize cells in concentrated suspensions. Here the cells are modeled as randomly orientated spheroidal particles with homogenous dielectric properties and suspended in turbid media. The inverse algorithm retrieves the geometrical parameters and the concentration of cells simultaneously by inverting the reduced scattering coefficient spectra obtained from multispectral diffuse optical tomography (MS-DOT). Both round and spheroidal cells are tested and the role of multiple and higher order scattering of particles on the performance of the algorithm is evaluated using different concentrations of cells. PMID:23372258

  16. Morphological characterization of cells in concentrated suspensions using multispectral diffuse optical tomography.

    PubMed

    Hajihashemi, Mohammad Reza; Li, Xiaoqi; Jiang, Huabei

    2012-10-01

    Based on a non-spherical model of particle scattering, we investigate the capabilities and limitations of a T-matrix based inverse algorithm to morphologically characterize cells in concentrated suspensions. Here the cells are modeled as randomly orientated spheroidal particles with homogenous dielectric properties and suspended in turbid media. The inverse algorithm retrieves the geometrical parameters and the concentration of cells simultaneously by inverting the reduced scattering coefficient spectra obtained from multispectral diffuse optical tomography (MS-DOT). Both round and spheroidal cells are tested and the role of multiple and higher order scattering of particles on the performance of the algorithm is evaluated using different concentrations of cells.

  17. Periodic Mesoporous Organosilica Nanoparticles with Controlled Morphologies and High Drug/Dye Loadings for Multicargo Delivery in Cancer Cells.

    PubMed

    Croissant, Jonas G; Fatieiev, Yevhen; Omar, Haneen; Anjum, Dalaver H; Gurinov, Andrey; Lu, Jie; Tamanoi, Fuyuhiko; Zink, Jeffrey I; Khashab, Niveen M

    2016-07-01

    Despite the worldwide interest generated by periodic mesoporous organosilica (PMO) bulk materials, the design of PMO nanomaterials with controlled morphology remains largely unexplored and their properties unknown. In this work, we describe the first study of PMO nanoparticles (NPs) based on meta-phenylene bridges, and we conducted a comparative structure-property relationship investigation with para-phenylene-bridged PMO NPs. Our findings indicate that the change of the isomer drastically affects the structure, morphology, size, porosity and thermal stability of PMO materials. We observed a much higher porosity and thermal stability of the para-based PMO which was likely due to a higher molecular periodicity. Additionally, the para isomer could generate multipodal NPs at very low stirring speed and upon this discovery we designed a phenylene-ethylene bridged PMO with a controlled Janus morphology. Unprecedentedly high payloads could be obtained from 40 to 110 wt % regardless of the organic bridge of PMOs. Finally, we demonstrate for the first time the co-delivery of two cargos by PMO NPs. Importantly, the cargo stability in PMOs did not require the capping of the pores, unlike pure silica, and the delivery could be autonomously triggered in cancer cells by acidic pH with nearly 70 % cell killing. PMID:27245497

  18. Methanol exposure interferes with morphological cell movements in the Drosophila embryo and causes increased apoptosis in the CNS.

    PubMed

    Mellerick, Dervla M; Liu, Heather

    2004-09-01

    Despite the significant contributions of tissue culture and bacterial models to toxicology, whole animal models for developmental neurotoxins are limited in availability and ease of experimentation. Because Drosophila is a well understood model for embryonic development that is highly accessible, we asked whether it could be used to study methanol developmental neurotoxicity. In the presence of 4% methanol, approximately 35% of embryos die and methanol exposure leads to severe CNS defects in about half those embryos, where the longitudinal connectives are dorsally displaced and commissure formation is severely reduced. In addition, a range of morphological defects in other germ layers is seen, and cell movement is adversely affected by methanol exposure. Although we did not find any evidence to suggest that methanol exposure affects the capacity of neuroblasts to divide or induces inappropriate apoptosis in these cells, in the CNS of germ band retracted embryos, the number of apoptotic nuclei is significantly increased in methanol-exposed embryos in comparison to controls, particularly in and adjacent to the ventral midline. Apoptosis contributes significantly to methanol neurotoxicity because embryos lacking the cell death genes grim, hid, and reaper have milder CNS defects resulting from methanol exposure than wild-type embryos. Our data suggest that when neurons and glia are severely adversely affected by methanol exposure, the damaged cells are cleared by apoptosis, leading to embryonic death. Thus, the Drosophila embryo may prove useful in identifying and unraveling mechanistic aspects of developmental neurotoxicity, specifically in relation to methanol toxicity.

  19. Image processing and classification algorithm for yeast cell morphology in a microfluidic chip

    NASA Astrophysics Data System (ADS)

    Yang Yu, Bo; Elbuken, Caglar; Ren, Carolyn L.; Huissoon, Jan P.

    2011-06-01

    The study of yeast cell morphology requires consistent identification of cell cycle phases based on cell bud size. A computer-based image processing algorithm is designed to automatically classify microscopic images of yeast cells in a microfluidic channel environment. The images were enhanced to reduce background noise, and a robust segmentation algorithm is developed to extract geometrical features including compactness, axis ratio, and bud size. The features are then used for classification, and the accuracy of various machine-learning classifiers is compared. The linear support vector machine, distance-based classification, and k-nearest-neighbor algorithm were the classifiers used in this experiment. The performance of the system under various illumination and focusing conditions were also tested. The results suggest it is possible to automatically classify yeast cells based on their morphological characteristics with noisy and low-contrast images.

  20. Human neural progenitors express functional lysophospholipid receptors that regulate cell growth and morphology

    PubMed Central

    Hurst, Jillian H; Mumaw, Jennifer; Machacek, David W; Sturkie, Carla; Callihan, Phillip; Stice, Steve L; Hooks, Shelley B

    2008-01-01

    Background Lysophospholipids regulate the morphology and growth of neurons, neural cell lines, and neural progenitors. A stable human neural progenitor cell line is not currently available in which to study the role of lysophospholipids in human neural development. We recently established a stable, adherent human embryonic stem cell-derived neuroepithelial (hES-NEP) cell line which recapitulates morphological and phenotypic features of neural progenitor cells isolated from fetal tissue. The goal of this study was to determine if hES-NEP cells express functional lysophospholipid receptors, and if activation of these receptors mediates cellular responses critical for neural development. Results Our results demonstrate that Lysophosphatidic Acid (LPA) and Sphingosine-1-phosphate (S1P) receptors are functionally expressed in hES-NEP cells and are coupled to multiple cellular signaling pathways. We have shown that transcript levels for S1P1 receptor increased significantly in the transition from embryonic stem cell to hES-NEP. hES-NEP cells express LPA and S1P receptors coupled to Gi/o G-proteins that inhibit adenylyl cyclase and to Gq-like phospholipase C activity. LPA and S1P also induce p44/42 ERK MAP kinase phosphorylation in these cells and stimulate cell proliferation via Gi/o coupled receptors in an Epidermal Growth Factor Receptor (EGFR)- and ERK-dependent pathway. In contrast, LPA and S1P stimulate transient cell rounding and aggregation that is independent of EGFR and ERK, but dependent on the Rho effector p160 ROCK. Conclusion Thus, lysophospholipids regulate neural progenitor growth and morphology through distinct mechanisms. These findings establish human ES cell-derived NEP cells as a model system for studying the role of lysophospholipids in neural progenitors. PMID:19077254

  1. Evaluation of the effects of Cimicifugae Rhizoma on the morphology and viability of mesenchymal stem cells

    PubMed Central

    JEONG, SU-HYEON; LEE, JI-EUN; KIM, BO-BAE; KO, YOUNGKYUNG; PARK, JUN-BEOM

    2015-01-01

    Cimicifugae Rhizoma is a traditional herbal medicine used to treat various diseases in Korea, China and Japan. Cimicifugae Rhizoma is primarily derived from Cimicifuga heracleifolia Komarov or Cimicifuga foetida Linnaeus. Cimicifugae Rhizoma has been used as an anti-inflammatory, analgesic and antipyretic remedy. The present study was performed to evaluate the extracts of Cimicifugae Rhizoma on the morphology and viability of human stem cells derived from gingiva. Stem cells derived from gingiva were grown in the presence of Cimicifugae Rhizoma at final concentrations that ranged from 0.001 to 1,000 µg/ml. The morphology of the cells was viewed under an inverted microscope and the analysis of cell proliferation was performed using a Cell Counting kit-8 (CCK-8) assay on days 1, 3, 5 and 7. Under an optical microscope, the control cells exhibited a spindle-shaped, fibroblast-like morphology. The shapes of the cells in the groups treated with 0.001, 0.01, 0.1, 1 and 10 µg/ml Cimicifugae Rhizoma were similar to the shapes in the control group. Significant alterations in morphology were noted in the 100 and 1,000 µg/ml groups when compared with the control group. The cells in the 100 and 1,000 µg/ml groups were rounder, and fewer cells were present. The cultures that were grown in the presence of Cimicifugae Rhizoma at a concentration of 0.001 µg/ml on day 1 had an increased CCK-8 value. The cultures grown in the presence of Cimicifugae Rhizoma at a concentration of 10 µg/ml on day 7 had a reduced CCK-8 value. Within the limits of this study, Cimicifugae Rhizoma influenced the viability of stem cells derived from the gingiva, and its direct application onto oral tissues may have adverse effects at high concentrations. The concentration and application time of Cimicifugae Rhizoma should be meticulously controlled to obtain optimal results. PMID:26622366

  2. Astrocytes grown in oculo: Expression of cell morphologies on the iris as revealed by GFA immunohistochemistry.

    PubMed

    Björklund, H

    1984-01-01

    Using two experimental approaches, the morphology of central astrocytes growing in vivo with the iris as a substratum were studied. When irides with mature intraocular grafts of cortex cerebri or locus coeruleus were stretch-prepared as whole mounts and processed for immunohistochemistry with antiserum against glial fibrillary acidic protein (GFA), a restricted halo of fluorescent cells and fibers was seen surrounding the grafts. Similarly, injection into the anterior eye chamber of adult rats, of a cell suspension prepared from cortex cerebri of 10-day-old rat pups gave rise to both multiple GFA-positive astrocytic islets of different sizes and cell densities as well as scattered individual cells on the anterior surface of the host iris. In contrast, astrocytes from similar cell suspensions prepared from young adult animals survived very poorly. In both types of experiments, a large variation in cell morphology ranging from immature epitheloid, via large flat cells with few thick processes, to typical mature star-shaped astrocytes was observed. This morphological variation is in agreement with that reported for similar cells in tissue culture. Immature-looking cells always had a strong perinuclear fluorescence; an inverse correlation was observed between cell body size and development of cell processes. Likewise, the fluorescence intensity was higher in well-developed cells as compared to more immature ones. The morphology of individual cells did not seem to be dependent upon the time in oculo, since no difference was observed between GFA-positive cells on irides examined 10 days and 6 weeks after injection of a cell suspension. Similarly, a high number of immature-looking cells was seen in irides with locus coeruleus transplants grafted more than 6 months earlier. Instead, the cell density seemed to be the crucial factor. Thus, star-shaped, well-developed cells were seen growing singly or in less dense groups whereas denser areas contained mainly immature

  3. Astrocytes grown in oculo: Expression of cell morphologies on the iris as revealed by GFA immunohistochemistry.

    PubMed

    Björklund, H

    1984-01-01

    Using two experimental approaches, the morphology of central astrocytes growing in vivo with the iris as a substratum were studied. When irides with mature intraocular grafts of cortex cerebri or locus coeruleus were stretch-prepared as whole mounts and processed for immunohistochemistry with antiserum against glial fibrillary acidic protein (GFA), a restricted halo of fluorescent cells and fibers was seen surrounding the grafts. Similarly, injection into the anterior eye chamber of adult rats, of a cell suspension prepared from cortex cerebri of 10-day-old rat pups gave rise to both multiple GFA-positive astrocytic islets of different sizes and cell densities as well as scattered individual cells on the anterior surface of the host iris. In contrast, astrocytes from similar cell suspensions prepared from young adult animals survived very poorly. In both types of experiments, a large variation in cell morphology ranging from immature epitheloid, via large flat cells with few thick processes, to typical mature star-shaped astrocytes was observed. This morphological variation is in agreement with that reported for similar cells in tissue culture. Immature-looking cells always had a strong perinuclear fluorescence; an inverse correlation was observed between cell body size and development of cell processes. Likewise, the fluorescence intensity was higher in well-developed cells as compared to more immature ones. The morphology of individual cells did not seem to be dependent upon the time in oculo, since no difference was observed between GFA-positive cells on irides examined 10 days and 6 weeks after injection of a cell suspension. Similarly, a high number of immature-looking cells was seen in irides with locus coeruleus transplants grafted more than 6 months earlier. Instead, the cell density seemed to be the crucial factor. Thus, star-shaped, well-developed cells were seen growing singly or in less dense groups whereas denser areas contained mainly immature

  4. Iron affects the structure of cell membrane molecular models.

    PubMed

    Suwalsky, M; Martínez, F; Cárdenas, H; Grzyb, J; Strzałka, K

    2005-03-01

    The effects of Fe(3+) and Fe(2+) on molecular models of biomembranes were investigated. These consisted of bilayers of dimyristoylphosphatidylcholine (DMPC) and of dimyristoylphosphatidylethanolamine (DMPE), classes of phospholipids located in the outer and inner moieties of cell membranes, respectively. X-ray studies showed that very low concentrations of Fe(3+) affected DMPC organization and 10(-3)M induced a total loss of its multilamellar periodic stacking. Experiments carried out with Fe(2+) on DMPC showed weaker effects than those induced by Fe(3+) ions. Similar experiments were performed on DMPE bilayers. Fe(3+) from 10(-7)M up to 10(-4)M had practically no effect on DMPE structure. However, 10(-3)M Fe(3+) induced a deep perturbation of the multilamellar structure of DMPE. However, 10(-3)M Fe(2+) had no effect on DMPE organization practically. Differential scanning calorimetry measurements also revealed different effects of Fe(3+) and Fe(2+) on the phase transition and other thermal properties of the examined lipids. In conclusion, the results obtained indicate that iron ions interact with phospholipid bilayers perturbing their structures. These findings are consistent with the observation that iron ions change cell membrane fluidity and, therefore, affect its functions. PMID:15752465

  5. Flavonoid-induced morphological modifications of endothelial cells through microtubule stabilization.

    PubMed

    Touil, Yasmine S; Fellous, Arlette; Scherman, Daniel; Chabot, Guy G

    2009-01-01

    Flavonoids are common components of the human diet and appear to be of interest in cancer prevention or therapy, but their structure-activity relationships (SAR) remain poorly defined. In this study, were compared 24 flavonoids for their cytotoxicity on cancer cells (B16 and Lewis lung) and their morphological effect on endothelial cells (EC) that could predict antiangiogenic activity. Ten flavonoids presented inhibitory concentrations for 50% of cancer cells (IC50, 48 h) below 50 microM: rhamnetin, 3',4'-dihydroxyflavone, luteolin, 3-hydroxyflavone, acacetin, apigenin, quercetin, baicalein, fisetin, and galangin. Important SAR for cytotoxicity included the C2-C3 double bond and 3',4'-dihydroxylation. Concerning the morphological effects on EC, only fisetin, quercetin, kaempferol, apigenin, and morin could induce the formation of cell extensions and filopodias at noncytotoxic concentrations. The SAR for morphologic activity differed from cytotoxicity and involved hydroxylation at C-7 and C-4'. Fisetin, the most active agent, presented cell morphology that was distinct compared to colchicine, combretastatin A-4, docetaxel, and cytochalasin D. Resistance to cold depolymerization and a 2.4-fold increase in acetylated alpha-tubulin demonstrated that fisetin was a microtubule stabilizer. In conclusion, this study disclosed several SAR that could guide the choice or the rational synthesis of improved flavonoids for cancer prevention or therapy.

  6. Flavonoid-induced morphological modifications of endothelial cells through microtubule stabilization

    PubMed Central

    Touil, Yasmine S.; Fellous, Arlette; Scherman, Daniel; Chabot, Guy G.

    2009-01-01

    Flavonoids are common components of the human diet and appear to be of interest in cancer prevention or therapy, but their structure-activity relationships (SAR) remain poorly defined. In this study, were compared 24 flavonoids for their cytotoxicity on cancer cells (B16 and Lewis lung), and their morphological effect on endothelial cells (EC) that could predict antiangiogenic activity. Ten flavonoids presented inhibitory concentrations for 50% of cancer cells (IC50, 48 h) below 50 μM: rhamnetin, 3′,4′-dihydroxyflavone, luteolin, 3-hydroxyflavone, acacetin, apigenin, quercetin, baicalein, fisetin, and galangin. Important SAR for cytotoxicity included the C2-C3 double bond and 3′,4′-dihydroxylation. Concerning the morphological effects on EC, only fisetin, quercetin, kaempferol, apigenin, and morin could induce the formation of cell extensions and filopodias at non cytotoxic concentrations. The SAR for morphologic activity differed from cytotoxicity and involved hydroxylation at C-7 and C-4′. Fisetin, the most active agent, presented cell morphology that was distinct compared to colchicine, combretastatin A-4, docetaxel, and cytochalasin D. Resistance to cold depolymerization and a 2.4-fold increase in acetylated α-tubulin demonstrated that fisetin was a microtubule stabilizer. In conclusion, this study disclosed several SAR that could guide the choice or the rational synthesis of improved flavonoids for cancer prevention or therapy. PMID:19373604

  7. Morphological Analysis of Cell Death by Cytospinning Followed by Rapid Staining.

    PubMed

    Crowley, Lisa C; Marfell, Brooke J; Waterhouse, Nigel J

    2016-01-01

    Identifying and characterizing different forms of cell death can be facilitated by staining internal cellular structures with dyes such as hematoxylin and eosin (H&E). These dyes stain the nucleus and cytoplasm, respectively, and optimized reagents (e.g., Rapi-Diff, Rapid Stain, or Quick Dip) are commonly used in pathology laboratories. Fixing and staining adherent cells with these optimized reagents is a straightforward procedure, but apoptotic cells may detach from the culture plate and be washed away during the fixing and staining procedure. To prevent the loss of apoptotic cells, cells can be gently centrifuged onto glass slides by cytospinning before fixing and staining. In addition to apoptotic cells, this procedure can be used on cells in suspension, or adherent cells that have been trypsinized and removed from the culture dish. This protocol describes cytospinning followed by Rapi-Diff staining for morphological analysis of cell death. PMID:27587773

  8. SB203580 promotes EGF-stimulated early morphological differentiation in PC12 cell through activating ERK pathway.

    PubMed

    New, L; Li, Y; Ge, B; Zhong, H; Mansbridge, J; Liu, K; Han, J

    2001-01-01

    MAP kinases have important role in PC12 cell differentiation, since the activities of both extracellular regulated protein kinase (ERK) and p38 have been indicated as necessary signal for PC12 cell differentiation. Epidermal growth factor (EGF) and NGF both activate ERK and p38 in PC12 cells, but only NGF trigger differentiation. It has been proposed that the duration of ERK activation determines the switch from proliferation to differentiation, since EGF causes more transient activation of ERK than NGF in PC12 cells. Here we report that treatment of PC12 cells with EGF in the presence of SB203580, a widely used p38 inhibitor, caused differentiation. The pro-differentiation effect of SB203580 in EGF-treated PC12 cells was found to be independent of its function of p38 inhibition but was through an effect on the ERK pathway that has been recently reported (Kalmes et al. [1999] FEBS Lett. 444: 71-74; Hall-Jackson et al. [1999] Onc. 18: 2047-2054). We found that SB203580 by itself did not affect the activity of ERK1/2 but significantly extended EGF-induced ERK activation in PC12 cells, which resulted in early morphological differentiation. Our data indicated that although both ERK and p38 are required for PC12 cell differentiation, activation of p38 is not required when ERK is superactivated. Our data provided further evidence for the threshold theory that differentiation is determined by the duration of ERK activation.

  9. Nanoporous gold membranes: From morphological control to fuel cell catalysis

    NASA Astrophysics Data System (ADS)

    Ding, Yi

    stable, low Pt usage, and better tolerance to CO poisoning. We incorporated it as a membrane electrode into a working proton exchange membrane fuel cells (PEMFC). Preliminary results show that Pt/NPG has very good fuel cell performance at a very low platinum loading.

  10. Genotype-environment interactions affecting preflowering physiological and morphological traits of Brassica rapa grown in two watering regimes.

    PubMed

    El-Soda, Mohamed; Boer, Martin P; Bagheri, Hedayat; Hanhart, Corrie J; Koornneef, Maarten; Aarts, Mark G M

    2014-02-01

    Plant growth and productivity are greatly affected by drought, which is likely to become more threatening with the predicted global temperature increase. Understanding the genetic architecture of complex quantitative traits and their interaction with water availability may lead to improved crop adaptation to a wide range of environments. Here, the genetic basis of 20 physiological and morphological traits is explored by describing plant performance and growth in a Brassica rapa recombinant inbred line (RIL) population grown on a sandy substrate supplemented with nutrient solution, under control and drought conditions. Altogether, 54 quantitative trait loci (QTL) were identified, of which many colocated in 11 QTL clusters. Seventeen QTL showed significant QTL-environment interaction (Q×E), indicating genetic variation for phenotypic plasticity. Of the measured traits, only hypocotyl length did not show significant genotype-environment interaction (G×E) in both environments in all experiments. Correlation analysis showed that, in the control environment, stomatal conductance was positively correlated with total leaf dry weight (DW) and aboveground DW, whereas in the drought environment, stomatal conductance showed a significant negative correlation with total leaf DW and aboveground DW. This correlation was explained by antagonistic fitness effects in the drought environment, controlled by a QTL cluster on chromosome A7. These results demonstrate that Q×E is an important component of the genetic variance and can play a great role in improving drought tolerance in future breeding programmes.

  11. Genotype-environment interactions affecting preflowering physiological and morphological traits of Brassica rapa grown in two watering regimes.

    PubMed

    El-Soda, Mohamed; Boer, Martin P; Bagheri, Hedayat; Hanhart, Corrie J; Koornneef, Maarten; Aarts, Mark G M

    2014-02-01

    Plant growth and productivity are greatly affected by drought, which is likely to become more threatening with the predicted global temperature increase. Understanding the genetic architecture of complex quantitative traits and their interaction with water availability may lead to improved crop adaptation to a wide range of environments. Here, the genetic basis of 20 physiological and morphological traits is explored by describing plant performance and growth in a Brassica rapa recombinant inbred line (RIL) population grown on a sandy substrate supplemented with nutrient solution, under control and drought conditions. Altogether, 54 quantitative trait loci (QTL) were identified, of which many colocated in 11 QTL clusters. Seventeen QTL showed significant QTL-environment interaction (Q×E), indicating genetic variation for phenotypic plasticity. Of the measured traits, only hypocotyl length did not show significant genotype-environment interaction (G×E) in both environments in all experiments. Correlation analysis showed that, in the control environment, stomatal conductance was positively correlated with total leaf dry weight (DW) and aboveground DW, whereas in the drought environment, stomatal conductance showed a significant negative correlation with total leaf DW and aboveground DW. This correlation was explained by antagonistic fitness effects in the drought environment, controlled by a QTL cluster on chromosome A7. These results demonstrate that Q×E is an important component of the genetic variance and can play a great role in improving drought tolerance in future breeding programmes. PMID:24474811

  12. Genotype–environment interactions affecting preflowering physiological and morphological traits of Brassica rapa grown in two watering regimes

    PubMed Central

    Aarts, Mark G. M.

    2014-01-01

    Plant growth and productivity are greatly affected by drought, which is likely to become more threatening with the predicted global temperature increase. Understanding the genetic architecture of complex quantitative traits and their interaction with water availability may lead to improved crop adaptation to a wide range of environments. Here, the genetic basis of 20 physiological and morphological traits is explored by describing plant performance and growth in a Brassica rapa recombinant inbred line (RIL) population grown on a sandy substrate supplemented with nutrient solution, under control and drought conditions. Altogether, 54 quantitative trait loci (QTL) were identified, of which many colocated in 11 QTL clusters. Seventeen QTL showed significant QTL–environment interaction (Q×E), indicating genetic variation for phenotypic plasticity. Of the measured traits, only hypocotyl length did not show significant genotype–environment interaction (G×E) in both environments in all experiments. Correlation analysis showed that, in the control environment, stomatal conductance was positively correlated with total leaf dry weight (DW) and aboveground DW, whereas in the drought environment, stomatal conductance showed a significant negative correlation with total leaf DW and aboveground DW. This correlation was explained by antagonistic fitness effects in the drought environment, controlled by a QTL cluster on chromosome A7. These results demonstrate that Q×E is an important component of the genetic variance and can play a great role in improving drought tolerance in future breeding programmes. PMID:24474811

  13. Dexamethasone and Azathioprine Promote Cytoskeletal Changes and Affect Mesenchymal Stem Cell Migratory Behavior

    PubMed Central

    Schneider, Natália; Gonçalves, Fabiany da Costa; Pinto, Fernanda Otesbelgue; Lopez, Patrícia Luciana da Costa; Araújo, Anelise Bergmann; Pfaffenseller, Bianca; Passos, Eduardo Pandolfi; Cirne-Lima, Elizabeth Obino; Meurer, Luíse; Lamers, Marcelo Lazzaron; Paz, Ana Helena

    2015-01-01

    Glucocorticoids and immunosuppressive drugs are commonly used to treat inflammatory disorders, such as inflammatory bowel disease (IBD), and despite a few improvements, the remission of IBD is still difficult to maintain. Due to their immunomodulatory properties, mesenchymal stem cells (MSCs) have emerged as regulators of the immune response, and their viability and activation of their migratory properties are essential for successful cell therapy. However, little is known about the effects of immunosuppressant drugs used in IBD treatment on MSC behavior. The aim of this study was to evaluate MSC viability, nuclear morphometry, cell polarity, F-actin and focal adhesion kinase (FAK) distribution, and cell migratory properties in the presence of the immunosuppressive drugs azathioprine (AZA) and dexamethasone (DEX). After an initial characterization, MSCs were treated with DEX (10 μM) or AZA (1 μM) for 24 hrs or 7 days. Neither drug had an effect on cell viability or nuclear morphometry. However, AZA treatment induced a more elongated cell shape, while DEX was associated with a more rounded cell shape (P < 0.05) with a higher presence of ventral actin stress fibers (P < 0.05) and a decrease in protrusion stability. After 7 days of treatment, AZA improved the cell spatial trajectory (ST) and increased the migration speed (24.35%, P < 0.05, n = 4), while DEX impaired ST and migration speed after 24 hrs and 7 days of treatment (-28.69% and -25.37%, respectively; P < 0.05, n = 4). In conclusion, our data suggest that these immunosuppressive drugs each affect MSC morphology and migratory capacity differently, possibly impacting the success of cell therapy. PMID:25756665

  14. Effects of Angular Frequency During Clinorotation on Mesenchymal Stem Cell Morphology and Migration

    NASA Technical Reports Server (NTRS)

    Luna, Carlos; Yew, Alvin G.; Hsieh, Adam H.

    2015-01-01

    Background/Objectives: Ground-based microgravity simulation can reproduce the apparent effects of weightlessness in spaceflight using clinostats that continuously reorient the gravity vector on a specimen, creating a time-averaged nullification of gravity. In this work, we investigated the effects of clinorotation speed on the morphology, cytoarchitecture, and migration behavior of human mesenchymal stem cells (hMSCs). Methods: We compared cell responses at clinorotation speeds of 0, 30, 60, and 75 rpm over 8 hours in a recently developed lab-on-chip-based clinostat system. Time lapse light microscopy was used to visualize changes in cell morphology during and after cessation of clinorotation. Cytoarchitecture was assessed by actin and vinculin staining, and chemotaxis was examined using time lapse light microscopy of cells in NGF (100 ng/ml) gradients. Results: Among clinorotated groups, cell area distributions indicated a greater inhibition of cell spreading with higher angular frequency (p is less than 0.005), though average cell area at 30 rpm after 8 hours became statistically similar to control (p = 0.794). Cells at 75rpm clinorotation remained viable and were able to re-spread after clinorotation. In chemotaxis chambers clinorotation did not alter migration patterns in elongated cells, but most clinorotated cells exhibited cell retraction, which strongly compromised motility.

  15. Morphological changes in human neural cells following tick-borne encephalitis virus infection.

    PubMed

    Růzek, Daniel; Vancová, Marie; Tesarová, Martina; Ahantarig, Arunee; Kopecký, Jan; Grubhoffer, Libor

    2009-07-01

    Tick-borne encephalitis (TBE) is one of the leading and most dangerous human viral neuroinfections in Europe and north-eastern Asia. The clinical manifestations include asymptomatic infections, fevers and debilitating encephalitis that might progress into chronic disease or fatal infection. To understand TBE pathology further in host nervous systems, three human neural cell lines, neuroblastoma, medulloblastoma and glioblastoma, were infected with TBE virus (TBEV). The susceptibility and virus-mediated cytopathic effect, including ultrastructural and apoptotic changes of the cells, were examined. All the neural cell lines tested were susceptible to TBEV infection. Interestingly, the neural cells produced about 100- to 10,000-fold higher virus titres than the conventional cell lines of extraneural origin, indicating the highly susceptible nature of neural cells to TBEV infection. The infection of medulloblastoma and glioblastoma cells was associated with a number of major morphological changes, including proliferation of membranes of the rough endoplasmic reticulum and extensive rearrangement of cytoskeletal structures. The TBEV-infected cells exhibited either necrotic or apoptotic morphological features. We observed ultrastructural apoptotic signs (condensation, margination and fragmentation of chromatin) and other alterations, such as vacuolation of the cytoplasm, dilatation of the endoplasmic reticulum cisternae and shrinkage of cells, accompanied by a high density of the cytoplasm. On the other hand, infected neuroblastoma cells did not exhibit proliferation of membranous structures. The virions were present in both the endoplasmic reticulum and the cytoplasm. Cells were dying preferentially by necrotic mechanisms rather than apoptosis. The neuropathological significance of these observations is discussed. PMID:19264624

  16. Suitable parameter choice on quantitative morphology of A549 cell in epithelial–mesenchymal transition

    PubMed Central

    Ren, Zhou-Xin; Yu, Hai-Bin; Li, Jian-Sheng; Shen, Jun-Ling; Du, Wen-Sen

    2015-01-01

    Evaluation of morphological changes in cells is an integral part of study on epithelial to mesenchymal transition (EMT), however, only a few papers reported the changes in quantitative parameters and no article compared different parameters for demanding better parameters. In the study, the purpose was to investigate suitable parameters for quantitative evaluation of EMT morphological changes. A549 human lung adenocarcinoma cell line was selected for the study. Some cells were stimulated by transforming growth factor-β1 (TGF-β1) for EMT, and other cells were as control without TGF-β1 stimulation. Subsequently, cells were placed in phase contrast microscope and three arbitrary fields were captured and saved with a personal computer. Using the tools of Photoshop software, some cells in an image were selected, segmented out and exchanged into unique hue, and other part in the image was shifted into another unique hue. The cells were calculated with 29 morphological parameters by Image Pro Plus software. A parameter between cells with or without TGF-β1 stimulation was compared statistically and nine parameters were significantly different between them. Receiver operating characteristic curve (ROC curve) of a parameter was described with SPSS software and F-test was used to compare two areas under the curves (AUCs) in Excel. Among them, roundness and radius ratio were the most AUCs and were significant higher than the other parameters. The results provided a new method with quantitative assessment of cell morphology during EMT, and found out two parameters, roundness and radius ratio, as suitable for quantification. PMID:26182364

  17. Morphology, properties, and performance of electrodeposited n-CdSe in liquid junction solar cells

    SciTech Connect

    Tomkiewicz, M.; Ling, I.; Parsons, W.S.

    1982-09-01

    The authors describe the mechanisms for galvanostatic electrodeposition of CdSe in terms of competition between chemical reactions that lead to Se formation and electrochemical reduction of Se as polyselenide, at the interfaces between selenium and selenide. This mechanism leads to a cauliflower morphology for the resulting film. This morphology is ideal for a photoanode in the liquid junction solar cell configuration, and the authors describe the performance of such an electrode. In spite of the unique morphology, solid-state properties of the film can be evaluated and the methodology for these evaluations is presented. The performance of the liquid junction solar cells is limited by the dark current and the dielectric properties of the material. The authors also describe the effects of metal ions such as Zn/sup +2/, Ru/sup +3/, and Ga/sup +3/ on the various electrode properties.

  18. Determining the optimum morphology in high-performance polymer-fullerene organic photovoltaic cells.

    PubMed

    Hedley, Gordon J; Ward, Alexander J; Alekseev, Alexander; Howells, Calvyn T; Martins, Emiliano R; Serrano, Luis A; Cooke, Graeme; Ruseckas, Arvydas; Samuel, Ifor D W

    2013-01-01

    The morphology of bulk heterojunction organic photovoltaic cells controls many of the performance characteristics of devices. However, measuring this morphology is challenging because of the small length-scales and low contrast between organic materials. Here we use nanoscale photocurrent mapping, ultrafast fluorescence and exciton diffusion to observe the detailed morphology of a high-performance blend of PTB7:PC71BM. We show that optimized blends consist of elongated fullerene-rich and polymer-rich fibre-like domains, which are 10-50 nm wide and 200-400 nm long. These elongated domains provide a concentration gradient for directional charge diffusion that helps in the extraction of charge pairs with 80% efficiency. In contrast, blends with agglomerated fullerene domains show a much lower efficiency of charge extraction of ~45%, which is attributed to poor electron and hole transport. Our results show that the formation of narrow and elongated domains is desirable for efficient bulk heterojunction solar cells.

  19. Monitoring cell morphology during necrosis and apoptosis by quantitative phase imaging

    NASA Astrophysics Data System (ADS)

    Mugnano, Martina; Calabuig, Alejandro; Grilli, Simonetta; Miccio, Lisa; Ferraro, Pietro

    2015-05-01

    Cellular morphology changes and volume alterations play significant roles in many biological processes and they are mirrors of cell functions. In this paper, we propose the Digital Holographic microscope (DH) as a non-invasive imaging technique for a rapid and accurate extraction of morphological information related to cell death. In particular, we investigate the morphological variations that occur during necrosis and apoptosis. The study of necrosis is extremely important because it is often associated with unwarranted loss of cells in human pathologies such as ischemia, trauma, and some forms of neurodegeneration; therefore, a better elucidation in terms of cell morphological changes could pave the way for new treatments. Also, apoptosis is extremely important because it's involved in cancer, both in its formation and in medical treatments. Because the inability to initiate apoptosis enhances tumour formation, current cancer treatments target this pathway. Within this framework, we have developed a transmission off-axis DH apparatus integrated with a micro incubator for investigation of living cells in a temperature and CO2 controlled environment. We employ DH to analyse the necrosis cell death induced by laser light (wavelength 473 nm, light power 4 mW). We have chosen as cellular model NIH 3T3 mouse embryonic fibroblasts because their adhesive features such as morphological changes, and the time needed to adhere and spread have been well characterized in the literature. We have monitored cell volume changes and morphological alterations in real time in order to study the necrosis process accurately and quantitatively. Cell volume changes were evaluated from the measured phase changes of light transmitted through cells. Our digital holographic experiments showed that after exposure of cells to laser light for 90-120 min., they swell and then take on a balloon-like shape until the plasma membrane ruptures and finally the cell volume decreases. Furthermore, we

  20. Influence of curvature on the morphology of brain microvascular endothelial cells

    NASA Astrophysics Data System (ADS)

    Ye, Mao; Yang, Zhen; Wong, Andrew; Searson, Peter; Searson Group Team

    2013-03-01

    There are hundreds or thousands of endothelial cells around the perimeter of a single artery or vein, and hence an individual cell experiences little curvature. In contrast, a single endothelial cell may wrap around itself to form the lumen of a brain capillary. Curvature plays a key role in many biological, chemical and physical processes, however, its role in dictating the morphology and polarization of brain capillary endothelial cells has not been investigated. We hypothesize that curvature and shear flow play a key role in determining the structure and function of the blood-brain barrier (BBB). We have developed the ``rod'' assay to study the influence of curvature on the morphology of confluent monolayers of endothelial cells. In this assay cells are plated onto glass rods pulled down to the desired diameter in the range from 5 - 500 μm and coated with collagen. We show that curvature has a significant influence on the morphology of endothelial cells and may have an important role in blood-brain barrier function.

  1. Changes in cell morphology due to plasma membrane wounding by acoustic cavitation

    PubMed Central

    Schlicher, Robyn K.; Hutcheson, Joshua D.; Radhakrishna, Harish; Apkarian, Robert P.; Prausnitz, Mark R.

    2010-01-01

    Acoustic cavitation-mediated wounding (i.e., sonoporation) has great potential to improve medical and laboratory applications requiring intracellular uptake of exogenous molecules; however, the field lacks detailed understanding of cavitation-induced morphological changes in cells and their relative importance. Here, we present an in-depth study of the effects of acoustic cavitation on cells using electron and confocal microscopy coupled with quantitative flow cytometry. High resolution images of treated cells show that morphologically different types of blebs can occur after wounding conditions caused by ultrasound exposure as well as by mechanical shear and strong laser ablation. In addition, these treatments caused wound-induced non-lytic necrotic death resulting in cell bodies we call wound-derived perikarya (WD-P). However, only cells exposed to acoustic cavitation experienced ejection of intact nuclei and nearly instant lytic necrosis. Quantitative analysis by flow cytometry indicates that wound-derived perikarya are the dominant morphology of nonviable cells, except at the strongest wounding conditions, where nuclear ejection accounts for a significant portion of cell death after ultrasound exposure. PMID:20350691

  2. Tricellulin deficiency affects tight junction architecture and cochlear hair cells

    PubMed Central

    Nayak, Gowri; Lee, Sue I.; Yousaf, Rizwan; Edelmann, Stephanie E.; Trincot, Claire; Van Itallie, Christina M.; Sinha, Ghanshyam P.; Rafeeq, Maria; Jones, Sherri M.; Belyantseva, Inna A.; Anderson, James M.; Forge, Andrew; Frolenkov, Gregory I.; Riazuddin, Saima

    2013-01-01

    The two compositionally distinct extracellular cochlear fluids, endolymph and perilymph, are separated by tight junctions that outline the scala media and reticular lamina. Mutations in TRIC (also known as MARVELD2), which encodes a tricellular tight junction protein known as tricellulin, lead to nonsyndromic hearing loss (DFNB49). We generated a knockin mouse that carries a mutation orthologous to the TRIC coding mutation linked to DFNB49 hearing loss in humans. Tricellulin was absent from the tricellular junctions in the inner ear epithelia of the mutant animals, which developed rapidly progressing hearing loss accompanied by loss of mechanosensory cochlear hair cells, while the endocochlear potential and paracellular permeability of a biotin-based tracer in the stria vascularis were unaltered. Freeze-fracture electron microscopy revealed disruption of the strands of intramembrane particles connecting bicellular and tricellular junctions in the inner ear epithelia of tricellulin-deficient mice. These ultrastructural changes may selectively affect the paracellular permeability of ions or small molecules, resulting in a toxic microenvironment for cochlear hair cells. Consistent with this hypothesis, hair cell loss was rescued in tricellulin-deficient mice when generation of normal endolymph was inhibited by a concomitant deletion of the transcription factor, Pou3f4. Finally, comprehensive phenotypic screening showed a broader pathological phenotype in the mutant mice, which highlights the non-redundant roles played by tricellulin. PMID:23979167

  3. Nuclear nano-morphology markers of histologically normal cells detect the "field effect" of breast cancer.

    PubMed

    Bista, Rajan K; Wang, Pin; Bhargava, Rohit; Uttam, Shikhar; Hartman, Douglas J; Brand, Randall E; Liu, Yang

    2012-08-01

    Accurate detection of breast malignancy from histologically normal cells ("field effect") has significant clinical implications in a broad base of breast cancer management, such as high-risk lesion management, personalized risk assessment, breast tumor recurrence, and tumor margin management. More accurate and clinically applicable tools to detect markers characteristic of breast cancer "field effect" that are able to guide the clinical management are urgently needed. We have recently developed a novel optical microscope, spatial-domain low-coherence quantitative phase microscopy, which extracts the nanoscale structural characteristics of cell nuclei (i.e., nuclear nano-morphology markers), using standard histology slides. In this proof-of-concept study, we present the use of these highly sensitive nuclear nano-morphology markers to identify breast malignancy from histologically normal cells. We investigated the nano-morphology markers from 154 patients with a broad spectrum of breast pathology entities, including normal breast tissue, non-proliferative benign lesions, proliferative lesions (without and with atypia), "malignant-adjacent" normal tissue, and invasive carcinoma. Our results show that the nuclear nano-morphology markers of "malignant-adjacent" normal tissue can detect the presence of invasive breast carcinoma with high accuracy and do not reflect normal aging. Further, we found that a progressive change in nuclear nano-morphology markers that parallel breast cancer risk, suggesting its potential use for risk stratification. These novel nano-morphology markers that detect breast cancerous changes from nanoscale structural characteristics of histologically normal cells could potentially benefit the diagnosis, risk assessment, prognosis, prevention, and treatment of breast cancer.

  4. Morphologic and proteomic characterization of exosomes released by cultured extravillous trophoblast cells

    SciTech Connect

    Atay, Safinur; Gercel-Taylor, Cicek; Kesimer, Mehmet; Taylor, Douglas D.

    2011-05-01

    Exosomes represent an important intercellular communication vehicle, mediating events essential for the decidual microenvironment. While we have demonstrated exosome induction of pro-inflammatory cytokines, to date, no extensive characterization of trophoblast-derived exosomes has been provided. Our objective was to provide a morphologic and proteomic characterization of these exosomes. Exosomes were isolated from the conditioned media of Swan71 human trophoblast cells by ultrafiltration and ultracentrifugation. These were analyzed for density (sucrose density gradient centrifugation), morphology (electron microscopy), size (dynamic light scattering) and protein composition (Ion Trap mass spectrometry and western immunoblotting). Based on density gradient centrifugation, microvesicles from Sw71 cells exhibit a density between 1.134 and 1.173 g/ml. Electron microscopy demonstrated that microvesicles from Sw71 cells exhibit the characteristic cup-shaped morphology of exosomes. Dynamic light scattering showed a bell-shaped curve, indicating a homogeneous population with a mean size of 165 nm {+-} 0.5 nm. Ion Trap mass spectrometry demonstrated the presence of exosome marker proteins (including CD81, Alix, cytoskeleton related proteins, and Rab family). The MS results were confirmed by western immunoblotting. Based on morphology, density, size and protein composition, we defined the release of exosomes from extravillous trophoblast cells and provide their first extensive characterization. This characterization is essential in furthering our understanding of 'normal' early pregnancy.

  5. Human aortic endothelial cell morphology influenced by topography of porous silicon substrates.

    PubMed

    Formentín, Pilar; Catalán, Úrsula; Fernández-Castillejo, Sara; Alba, Maria; Baranowska, Malgorzata; Solà, Rosa; Pallarès, Josep; Marsal, Lluís F

    2015-10-01

    Porous silicon has received much attention because of its optical properties and for its usefulness in cell-based biosensing, drug delivery, and tissue engineering applications. Surface properties of the biomaterial are associated with cell adhesion and with proliferation, migration, and differentiation. The present article analyzes the behavior of human aortic endothelial cells in macro- and nanoporous collagen-modified porous silicon samples. On both substrates, cells are well adhered and numerous. Confocal microscopy and scanning electron microscopy were employed to study the effects of porosity on the morphology of the cells. On macroporous silicon, filopodia is not observed but the cell spreads on the surface, increasing the lamellipodia surface which penetrates the macropore. On nanoporous silicon, multiple filopodia were found to branch out from the cell body. These results demonstrate that the pore size plays a key role in controlling the morphology and growth rate of human aortic endothelial cells, and that these forms of silicon can be used to control cell development in tissue engineering as well as in basic cell biology research.

  6. Acetate Salts as Nonhalogen Additives To Improve Perovskite Film Morphology for High-Efficiency Solar Cells.

    PubMed

    Wu, Qiliang; Zhou, Pengcheng; Zhou, Weiran; Wei, Xiangfeng; Chen, Tao; Yang, Shangfeng

    2016-06-22

    A two-step method has been popularly adopted to fabricate a perovskite film of planar heterojunction organo-lead halide perovskite solar cells (PSCs). However, this method often generates uncontrollable film morphology with poor coverage. Herein, we report a facile method to improve perovskite film morphology by incorporating a small amount of acetate (CH3COO(-), Ac(-)) salts (NH4Ac, NaAc) as nonhalogen additives in CH3NH3I solution used for immersing PbI2 film, resulting in improved CH3NH3PbI3 film morphology. Under the optimized NH4Ac additive concentration of 10 wt %, the best power conversion efficiency (PCE) reaches 17.02%, which is enhanced by ∼23.2% relative to that of the pristine device without additive, whereas the NaAc additive does not lead to an efficiency enhancement despite the improvement of the CH3NH3PbI3 film morphology. SEM study reveals that NH4Ac and NaAc additives can both effectively improve perovskite film morphology by increasing the surface coverage via diminishing pinholes. The improvement on CH3NH3PbI3 film morphology is beneficial for increasing the optical absorption of perovskite film and improving the interfacial contact at the perovskite/spiro-OMeTAD interface, leading to the increase of short-circuit current and consequently efficiency enhancement of the PSC device for NH4Ac additive only.

  7. Modifications in astrocyte morphology and calcium signaling induced by a brain capillary endothelial cell line.

    PubMed

    Yoder, Elizabeth J

    2002-04-15

    Astrocytes extend specialized endfoot processes to perisynaptic and perivascular regions, and thus are positioned to mediate the bidirectional flow of metabolic, ionic, and other transmissive substances between neurons and the blood stream. While mutual structural and functional interactions between neurons and astrocytes have been documented, less is known about the interactions between astrocytes and cerebrovascular cells. For example, although the ability of astrocytes to induce structural and functional changes in endothelial cells is established, the reciprocity of brain endothelial cells to induce changes in astrocytes is undetermined. This issue is addressed in the present study. Changes in primary cultures of neonatal mouse cortical astrocytes were investigated following their coculture with mouse brain capillary endothelial (bEnd3) cells. The presence of bEnd3 cells altered the morphology of astrocytes by transforming them from confluent monolayers into networks of elongated multicellular columns. These columns did not occur when either bEnd3 cells or astrocytes were cocultured with other cell types, suggesting that astrocytes undergo specific morphological consequences when placed in close proximity to brain endothelial cells. In addition to these structural changes, the pharmacological profile of astrocytes was modified by coculture with bEnd3 cells. Astrocytes in the cocultures showed an increased Ca2+ responsiveness to bradykinin and glutamate, but no change in responsiveness to ATP, as compared to controls. Coculturing the astrocytes with a neuronal cell line resulted in increased responsiveness of the glial responses to glutamate but not to bradykinin. These studies indicate that brain endothelial cells induce changes in astrocyte morphology and pharmacology.

  8. Modifications in astrocyte morphology and calcium signaling induced by a brain capillary endothelial cell line.

    PubMed

    Yoder, Elizabeth J

    2002-04-15

    Astrocytes extend specialized endfoot processes to perisynaptic and perivascular regions, and thus are positioned to mediate the bidirectional flow of metabolic, ionic, and other transmissive substances between neurons and the blood stream. While mutual structural and functional interactions between neurons and astrocytes have been documented, less is known about the interactions between astrocytes and cerebrovascular cells. For example, although the ability of astrocytes to induce structural and functional changes in endothelial cells is established, the reciprocity of brain endothelial cells to induce changes in astrocytes is undetermined. This issue is addressed in the present study. Changes in primary cultures of neonatal mouse cortical astrocytes were investigated following their coculture with mouse brain capillary endothelial (bEnd3) cells. The presence of bEnd3 cells altered the morphology of astrocytes by transforming them from confluent monolayers into networks of elongated multicellular columns. These columns did not occur when either bEnd3 cells or astrocytes were cocultured with other cell types, suggesting that astrocytes undergo specific morphological consequences when placed in close proximity to brain endothelial cells. In addition to these structural changes, the pharmacological profile of astrocytes was modified by coculture with bEnd3 cells. Astrocytes in the cocultures showed an increased Ca2+ responsiveness to bradykinin and glutamate, but no change in responsiveness to ATP, as compared to controls. Coculturing the astrocytes with a neuronal cell line resulted in increased responsiveness of the glial responses to glutamate but not to bradykinin. These studies indicate that brain endothelial cells induce changes in astrocyte morphology and pharmacology. PMID:11948807

  9. mGluR5 Positive and Negative Allosteric Modulators Differentially Affect Dendritic Spine Density and Morphology in the Prefrontal Cortex.

    PubMed

    LaCrosse, Amber L; Taylor, Sara B; Nemirovsky, Natali E; Gass, Justin T; Olive, Michael F

    2015-01-01

    Positive and negative allosteric modulators (PAMs and NAMs, respectively) of type 5 metabotropic glutamate receptors (mGluR5) are currently being investigated as novel treatments for neuropsychiatric diseases including drug addiction, schizophrenia, and Fragile X syndrome. However, only a handful of studies have examined the effects of mGluR5 PAMs or NAMs on the structural plasticity of dendritic spines in otherwise naïve animals, particularly in brain regions mediating executive function. In the present study, we assessed dendritic spine density and morphology in pyramidal cells of the medial prefrontal cortex (mPFC) after repeated administration of either the prototypical mGluR5 PAM 3-cyano-N-(1,3-diphenyl-1H-pyrazol-5- yl)benzamide (CDPPB, 20 mg/kg), the clinically utilized mGluR5 NAM 1-(3-chlorophenyl)-3-(3-methyl-5-oxo-4Himidazol- 2-yl)urea (fenobam, 20 mg/kg), or vehicle in male Sprague-Dawley rats. Following once daily treatment for 10 consecutive days, coronal brain sections containing the mPFC underwent diolistic labeling and 3D image analysis of dendritic spines. Compared to vehicle treated animals, rats administered fenobam exhibited significant increases in dendritic spine density and the overall frequency of spines with small (<0.2 μm) head diameters, decreases in frequency of spines with medium (0.2-0.4 μm) head diameters, and had no changes in frequency of spines with large head diameters (>0.4 μm). Administration of CDPPB had no discernable effects on dendritic spine density or morphology, and neither CDPPB nor fenobam had any effect on spine length or volume. We conclude that mGluR5 PAMs and NAMs differentially affect mPFC dendritic spine structural plasticity in otherwise naïve animals, and additional studies assessing their effects in combination with cognitive or behavioral tasks are needed.

  10. Acinic cell carcinoma of breast: morphologic and immunohistochemical review of a rare breast cancer subtype☆

    PubMed Central

    Conlon, Niamh; Sadri, Navid; Corben, Adriana D.; Tan, Lee K.

    2016-01-01

    Summary Acinic cell carcinoma of breast is a rare subtype of triple-negative breast carcinoma and demonstrates extensive morphologic overlap with acinic cell carcinoma of the salivary gland. In this study, we perform a detailed morphologic and immunohistochemical description of 2 cases of this rare entity and undertake a comprehensive review of all reported cases of breast acinic cell carcinoma in the English language literature to date. One-third of reported cases of breast acinic cell carcinoma have been associated with the presence of a ductal carcinoma not otherwise specified component, which is frequently poorly differentiated. Breast acinic cell carcinoma can demonstrate focal morphologic features similar to microglandular adenosis; these areas are frequently negative for collagen IV and laminin on immunohistochemistry. The true relationship between these 2 entities remains unclear, but we advocate that microglandular adenosis–like areas at the periphery of a breast acinic cell carcinoma should be considered part of the carcinomatous process and re-excised if this process extends to the initial surgical margins. PMID:27067778

  11. Genetic backgrounds and redox conditions influence morphological characteristics and cell differentiation of osteoclasts in mice.

    PubMed

    Narahara, Shun; Matsushima, Haruna; Sakai, Eiko; Fukuma, Yutaka; Nishishita, Kazuhisa; Okamoto, Kuniaki; Tsukuba, Takayuki

    2012-04-01

    Osteoclasts (OCLs) are multinucleated giant cells and are formed by the fusion of mononuclear progenitors of monocyte/macrophage lineage. It is known that macrophages derived from different genetic backgrounds exhibit quite distinct characteristics of immune responses. However, it is unknown whether OCLs from different genetic backgrounds show distinct characteristics. In this study, we showed that bone-marrow macrophages (BMMs) derived from C57BL/6, BALB/c and ddY mice exhibited considerably distinct morphological characteristics and cell differentiation into OCLs. The differentiation of BMMs into OCLs was comparatively quicker in the C57BL/6 and ddY mice, while that of BALB/c mice was rather slow. Morphologically, ddY OCLs showed a giant cell with a round shape, C57BL/6 OCLs were of a moderate size with many protrusions and BALB/c OCLs had the smallest size with fewer nuclei. The intracellular signaling of differentiation and expression levels of marker proteins of OCLs were different in the respective strains. Treatment of BMMs from the three different strains with the reducing agent N-acetylcysteine (NAC) or with the oxidation agent hydrogen peroxide (H(2)O(2)) induced changes in the shape and sizes of the cells and caused distinct patterns of cell differentiation and survival. Thus, genetic backgrounds and redox conditions regulate the morphological characteristics and cell differentiation of OCLs.

  12. Acinic cell carcinoma of breast: morphologic and immunohistochemical review of a rare breast cancer subtype.

    PubMed

    Conlon, Niamh; Sadri, Navid; Corben, Adriana D; Tan, Lee K

    2016-05-01

    Acinic cell carcinoma of breast is a rare subtype of triple-negative breast carcinoma and demonstrates extensive morphologic overlap with acinic cell carcinoma of the salivary gland. In this study, we perform a detailed morphologic and immunohistochemical description of 2 cases of this rare entity and undertake a comprehensive review of all reported cases of breast acinic cell carcinoma in the English language literature to date. One-third of reported cases of breast acinic cell carcinoma have been associated with the presence of a ductal carcinoma not otherwise specified component, which is frequently poorly differentiated. Breast acinic cell carcinoma can demonstrate focal morphologic features similar to microglandular adenosis; these areas are frequently negative for collagen IV and laminin on immunohistochemistry. The true relationship between these 2 entities remains unclear, but we advocate that microglandular adenosis-like areas at the periphery of a breast acinic cell carcinoma should be considered part of the carcinomatous process and re-excised if this process extends to the initial surgical margins. PMID:27067778

  13. Hybrid morphology dependence of CdTe:CdSe bulk-heterojunction solar cells.

    PubMed

    Tan, Furui; Qu, Shengchun; Zhang, Weifeng; Wang, Zhanguo

    2014-01-01

    A nanocrystal thin-film solar cell operating on an exciton splitting pattern requires a highly efficient separation of electron-hole pairs and transportation of separated charges. A hybrid bulk-heterojunction (HBH) nanostructure providing a large contact area and interpenetrated charge channels is favorable to an inorganic nanocrystal solar cell with high performance. For this freshly appeared structure, here in this work, we have firstly explored the influence of hybrid morphology on the photovoltaic performance of CdTe:CdSe bulk-heterojunction solar cells with variation in CdSe nanoparticle morphology. Quantum dot (QD) or nanotetrapod (NT)-shaped CdSe nanocrystals have been employed together with CdTe NTs to construct different hybrid structures. The solar cells with the two different hybrid active layers show obvious difference in photovoltaic performance. The hybrid structure with densely packed and continuously interpenetrated two phases generates superior morphological and electrical properties for more efficient inorganic bulk-heterojunction solar cells, which could be readily realized in the NTs:QDs hybrid. This proved strategy is applicable and promising in designing other highly efficient inorganic hybrid solar cells.

  14. Mucinous spindle and tubular renal cell carcinoma: analysis of chromosomal aberration pattern of low-grade, high-grade, and overlapping morphologic variant with papillary renal cell carcinoma.

    PubMed

    Peckova, Kvetoslava; Martinek, Petr; Sperga, Maris; Montiel, Delia Perez; Daum, Ondrej; Rotterova, Pavla; Kalusová, Kristýna; Hora, Milan; Pivovarcikova, Kristýna; Rychly, Boris; Vranic, Semir; Davidson, Whitney; Vodicka, Josef; Dubová, Magdaléna; Michal, Michal; Hes, Ondrej

    2015-08-01

    The chromosomal numerical aberration pattern in mucinous tubular and spindle renal cell carcinoma (MTSRCC) is referred to as variable with frequent gains and losses. The objectives of this study are to map the spectrum of chromosomal aberrations (extent and location) in a large cohort of the cases and relate these findings to the morphologic variants of MTSRCC. Fifty-four MTSRCCs with uniform morphologic pattern were selected (of 133 MTSRCCs available in our registry) and divided into 3 groups: classic low-grade MTSRCC (Fuhrman nucleolar International Society of Urological Pathology grade 2), high-grade MTSRCC (grade 3), and overlapping MTSRCC with papillary renal cell carcinoma (RCC) morphology. Array comparative genomic hybridization analysis was applied to 16 cases in which DNA was well preserved. Four analyzable classic low-grade MTSRCCs showed multiple losses affecting chromosomes 1, 4, 8, 9, 14, 15, and 22. No chromosomal gains were found. Four analyzable cases of MTSRCC showing overlapping morphology with PRCC displayed a more variable pattern including normal chromosomal status; losses of chromosomes 1, 6, 8, 9, 14, 15, and 22; and gains of 3, 7, 16, and 17. The group of 4 high-grade MTSRCCs exhibited a more uniform chromosomal aberration pattern with losses of chromosomes 1, 4, 6, 8, 9, 13, 14, 15, and 22 and without any gains detected. (1) MTSRCC, both low-grade and high-grade, shows chromosomal losses (including 1, 4, 6, 8, 9, 13, 14, 15, and 22) in all analyzable cases; this seems to be the most frequent chromosomal numerical aberration in this type of RCC. (2) Cases with overlapping morphologic features (MTSRCC and PRCC) showed a more variable pattern with multiple losses and gains, including gains of chromosomes 7 and 17 (2 cases). This result is in line with previously published morphologic and immunohistochemical studies that describe the broad morphologic spectrum of MTSRCC, with changes resembling papillary RCC. (3) The diagnosis of MTSRCC in

  15. Resolving Tumor Heterogeneity: Genes Involved in Chordoma Cell Development Identified by Low-Template Analysis of Morphologically Distinct Cells

    PubMed Central

    Wagner, Karin; Meditz, Katharina; Kolb, Dagmar; Feichtinger, Julia; Thallinger, Gerhard G.; Quehenberger, Franz; Liegl-Atzwanger, Bernadette; Rinner, Beate

    2014-01-01

    The classical sacrococcygeal chordoma tumor presents with a typical morphology of lobulated myxoid tumor tissue with cords, strands and nests of tumor cells. The population of cells consists of small non-vacuolated cells, intermediate cells with a wide range of vacuolization and large heavily vacuolated (physaliferous) cells. To date analysis was only performed on bulk tumor mass because of its rare incidence, lack of suited model systems and technical limitations thereby neglecting its heterogeneous composition. We intended to clarify whether the observed cell types are derived from genetically distinct clones or represent different phenotypes. Furthermore, we aimed at elucidating the differences between small non-vacuolated and large physaliferous cells on the genomic and transcriptomic level. Phenotype-specific analyses of small non-vacuolated and large physaliferous cells in two independent chordoma cell lines yielded four candidate genes involved in chordoma cell development. UCHL3, coding for an ubiquitin hydrolase, was found to be over-expressed in the large physaliferous cell phenotype of MUG-Chor1 (18.7-fold) and U-CH1 (3.7-fold) cells. The mannosyltransferase ALG11 (695-fold) and the phosphatase subunit PPP2CB (18.6-fold) were found to be up-regulated in large physaliferous MUG-Chor1 cells showing a similar trend in U-CH1 cells. TMEM144, an orphan 10-transmembrane family receptor, yielded contradictory data as cDNA microarray analysis showed up- but RT-qPCR data down-regulation in large physaliferous MUG-Chor1 cells. Isolation of few but morphologically identical cells allowed us to overcome the limitations of bulk analysis in chordoma research. We identified the different chordoma cell phenotypes to be part of a developmental process and discovered new genes linked to chordoma cell development representing potential targets for further research in chordoma tumor biology. PMID:24503940

  16. Effects of temperature and cellular interactions on the mechanics and morphology of human cancer cells investigated by atomic force microscopy.

    PubMed

    Li, Mi; Liu, LianQing; Xi, Ning; Wang, YueChao; Xiao, XiuBin; Zhang, WeiJing

    2015-09-01

    Cell mechanics plays an important role in cellular physiological activities. Recent studies have shown that cellular mechanical properties are novel biomarkers for indicating the cell states. In this article, temperature-controllable atomic force microscopy (AFM) was applied to quantitatively investigate the effects of temperature and cellular interactions on the mechanics and morphology of human cancer cells. First, AFM indenting experiments were performed on six types of human cells to investigate the changes of cellular Young's modulus at different temperatures and the results showed that the mechanical responses to the changes of temperature were variable for different types of cancer cells. Second, AFM imaging experiments were performed to observe the morphological changes in living cells at different temperatures and the results showed the significant changes of cell morphology caused by the alterations of temperature. Finally, by co-culturing human cancer cells with human immune cells, the mechanical and morphological changes in cancer cells were investigated. The results showed that the co-culture of cancer cells and immune cells could cause the distinct mechanical changes in cancer cells, but no significant morphological differences were observed. The experimental results improved our understanding of the effects of temperature and cellular interactions on the mechanics and morphology of cancer cells.

  17. Accurate Morphology Preserving Segmentation of Overlapping Cells based on Active Contours.

    PubMed

    Molnar, Csaba; Jermyn, Ian H; Kato, Zoltan; Rahkama, Vesa; Östling, Päivi; Mikkonen, Piia; Pietiäinen, Vilja; Horvath, Peter

    2016-01-01

    The identification of fluorescently stained cell nuclei is the basis of cell detection, segmentation, and feature extraction in high content microscopy experiments. The nuclear morphology of single cells is also one of the essential indicators of phenotypic variation. However, the cells used in experiments can lose their contact inhibition, and can therefore pile up on top of each other, making the detection of single cells extremely challenging using current segmentation methods. The model we present here can detect cell nuclei and their morphology even in high-confluency cell cultures with many overlapping cell nuclei. We combine the "gas of near circles" active contour model, which favors circular shapes but allows slight variations around them, with a new data model. This captures a common property of many microscopic imaging techniques: the intensities from superposed nuclei are additive, so that two overlapping nuclei, for example, have a total intensity that is approximately double the intensity of a single nucleus. We demonstrate the power of our method on microscopic images of cells, comparing the results with those obtained from a widely used approach, and with manual image segmentations by experts. PMID:27561654

  18. Accurate Morphology Preserving Segmentation of Overlapping Cells based on Active Contours

    PubMed Central

    Molnar, Csaba; Jermyn, Ian H.; Kato, Zoltan; Rahkama, Vesa; Östling, Päivi; Mikkonen, Piia; Pietiäinen, Vilja; Horvath, Peter

    2016-01-01

    The identification of fluorescently stained cell nuclei is the basis of cell detection, segmentation, and feature extraction in high content microscopy experiments. The nuclear morphology of single cells is also one of the essential indicators of phenotypic variation. However, the cells used in experiments can lose their contact inhibition, and can therefore pile up on top of each other, making the detection of single cells extremely challenging using current segmentation methods. The model we present here can detect cell nuclei and their morphology even in high-confluency cell cultures with many overlapping cell nuclei. We combine the “gas of near circles” active contour model, which favors circular shapes but allows slight variations around them, with a new data model. This captures a common property of many microscopic imaging techniques: the intensities from superposed nuclei are additive, so that two overlapping nuclei, for example, have a total intensity that is approximately double the intensity of a single nucleus. We demonstrate the power of our method on microscopic images of cells, comparing the results with those obtained from a widely used approach, and with manual image segmentations by experts. PMID:27561654

  19. Vaccinia virus F5 is required for normal plaque morphology in multiple cell lines but not replication in culture or virulence in mice.

    PubMed

    Dobson, Bianca M; Procter, Dean J; Hollett, Natasha A; Flesch, Inge E A; Newsome, Timothy P; Tscharke, David C

    2014-05-01

    Vaccinia virus (VACV) gene F5L was recently identified as a determinant of plaque morphology that is truncated in Modified Vaccinia virus Ankara (MVA). Here we show that F5L also affects plaque morphology of the virulent VACV strain Western Reserve (WR) in some, but not all cell lines, and not via previously described mechanisms. Further, despite a reduction in plaque size for VACV WR lacking F5L there was no evidence of reduced virus replication or spread in vitro or in vivo. In vivo we examined two mouse models, each with more than one dose and measured signs of disease and virus burden. These data provide an initial characterization of VACV F5L in a virulent strain of VACV. Further they show the necessity of testing plaque phenotypes in more than one cell type and provide an example of a VACV gene required for normal plaque morphology but not replication and spread.

  20. Effects of tacrolimus on morphology, proliferation and differentiation of mesenchymal stem cells derived from gingiva tissue

    PubMed Central

    HA, DONG-HO; YONG, CHUL SOON; KIM, JONG OH; JEONG, JEE-HEON; PARK, JUN-BEOM

    2016-01-01

    Tacrolimus is a 23-membered macrolide lactone with potent immunosuppressive activity that is effective in the prophylaxis of organ rejection following kidney, heart and liver transplantation. Tacrolimus also exerts a variety of actions on bone metabolism. The aim of the present study was to evaluate the effects of different concentrations of tacrolimus on the morphology and viability of human stem cells derived from the gingiva. Gingival-derived stem cells were grown in the presence of tacrolimus at final concentrations ranging from 0.001 to 100 µg/ml. The morphology of the cells was viewed under an inverted microscope and the cell viability was analyzed using Cell Counting kit-8 (CCK-8) on days 1, 3, 5 and 7. Alizarin Red S staining was used to assess mineralization of treated cells. The control group showed spindle-shaped, fibroblast-like morphology and the shapes of the cells in 0.001, 0.01, 0.1, 1 and 10 µg/ml tacrolimus were similar to those of the control group. All groups except the 100 µg/ml group showed increased cell proliferation over time. Cultures grown in the presence of tacrolimus at 0.001, 0.01, 0.1, 1 and 10 µg/ml were not identified to be significantly different compared with the control at days 1, 3 and 5 using the CCK-8 assays. Increased mineralized deposits were noted with increased incubation time. Treatment with tacrolimus from 0.001 to 1 µg/ml led to an increase in mineralization compared with the control group. Within the limits of this study, tacrolimus at the tested concentrations (ranging from 0.001 to 10 µg/ml) did not result in differences in the viability of stem cells derived from gingiva; however it did enhance osteogenic differentiation of the stem cells. PMID:27177273

  1. Effects of tacrolimus on morphology, proliferation and differentiation of mesenchymal stem cells derived from gingiva tissue.

    PubMed

    Ha, Dong-Ho; Yong, Chul Soon; Kim, Jong Oh; Jeong, Jee-Heon; Park, Jun-Beom

    2016-07-01

    Tacrolimus is a 23-membered macrolide lactone with potent immunosuppressive activity that is effective in the prophylaxis of organ rejection following kidney, heart and liver transplantation. Tacrolimus also exerts a variety of actions on bone metabolism. The aim of the present study was to evaluate the effects of different concentrations of tacrolimus on the morphology and viability of human stem cells derived from the gingiva. Gingival‑derived stem cells were grown in the presence of tacrolimus at final concentrations ranging from 0.001 to 100 µg/ml. The morphology of the cells was viewed under an inverted microscope and the cell viability was analyzed using Cell Counting kit‑8 (CCK‑8) on days 1, 3, 5 and 7. Alizarin Red S staining was used to assess mineralization of treated cells. The control group showed spindle‑shaped, fibroblast‑like morphology and the shapes of the cells in 0.001, 0.01, 0.1, 1 and 10 µg/ml tacrolimus were similar to those of the control group. All groups except the 100 µg/ml group showed increased cell proliferation over time. Cultures grown in the presence of tacrolimus at 0.001, 0.01, 0.1, 1 and 10 µg/ml were not identified to be significantly different compared with the control at days 1, 3 and 5 using the CCK‑8 assays. Increased mineralized deposits were noted with increased incubation time. Treatment with tacrolimus from 0.001 to 1 µg/ml led to an increase in mineralization compared with the control group. Within the limits of this study, tacrolimus at the tested concentrations (ranging from 0.001 to 10 µg/ml) did not result in differences in the viability of stem cells derived from gingiva; however it did enhance osteogenic differentiation of the stem cells.

  2. Immerse precipitation as an efficient protocol to optimize morphology and performance of organic solar cells

    NASA Astrophysics Data System (ADS)

    Zuo, Lijian; Cao, Zhen; Hu, Xiaolian; Gu, Zhuowei; Pan, Hongbin; Chen, Hongzheng

    2012-12-01

    We developed a film-forming processing method for morphology control and organic solar cells (OSCs) optimization. In this protocol, the processing solvent inside a wet active layer is removed by dripping a soaking solvent that is selectively soluble for the processing solvents onto the wet active layer film. By this method, OSCs based on diketopyrrolopyrrole containing copolymers: [6,6]-phenyl-C61-butyric acid methyl ester blend were fabricated. Devices processed by this treatment show a significantly increased efficiency by a factor of 3 compared to devices fabricated by the traditional spin-coating method (from 1.03% to 3.2%), which is mainly attributed to morphology improvements.

  3. New common variants affecting susceptibility to basal cell carcinoma.

    PubMed

    Stacey, Simon N; Sulem, Patrick; Masson, Gisli; Gudjonsson, Sigurjon A; Thorleifsson, Gudmar; Jakobsdottir, Margret; Sigurdsson, Asgeir; Gudbjartsson, Daniel F; Sigurgeirsson, Bardur; Benediktsdottir, Kristrun R; Thorisdottir, Kristin; Ragnarsson, Rafn; Scherer, Dominique; Hemminki, Kari; Rudnai, Peter; Gurzau, Eugene; Koppova, Kvetoslava; Botella-Estrada, Rafael; Soriano, Virtudes; Juberias, Pablo; Saez, Berta; Gilaberte, Yolanda; Fuentelsaz, Victoria; Corredera, Cristina; Grasa, Matilde; Höiom, Veronica; Lindblom, Annika; Bonenkamp, Johannes J; van Rossum, Michelle M; Aben, Katja K H; de Vries, Esther; Santinami, Mario; Di Mauro, Maria G; Maurichi, Andrea; Wendt, Judith; Hochleitner, Pia; Pehamberger, Hubert; Gudmundsson, Julius; Magnusdottir, Droplaug N; Gretarsdottir, Solveig; Holm, Hilma; Steinthorsdottir, Valgerdur; Frigge, Michael L; Blondal, Thorarinn; Saemundsdottir, Jona; Bjarnason, Hjördis; Kristjansson, Kristleifur; Bjornsdottir, Gyda; Okamoto, Ichiro; Rivoltini, Licia; Rodolfo, Monica; Kiemeney, Lambertus A; Hansson, Johan; Nagore, Eduardo; Mayordomo, José I; Kumar, Rajiv; Karagas, Margaret R; Nelson, Heather H; Gulcher, Jeffrey R; Rafnar, Thorunn; Thorsteinsdottir, Unnur; Olafsson, Jon H; Kong, Augustine; Stefansson, Kari

    2009-08-01

    In a follow-up to our previously reported genome-wide association study of cutaneous basal cell carcinoma (BCC), we describe here several new susceptibility variants. SNP rs11170164, encoding a G138E substitution in the keratin 5 (KRT5) gene, affects risk of BCC (OR = 1.35, P = 2.1 x 10(-9)). A variant at 9p21 near CDKN2A and CDKN2B also confers susceptibility to BCC (rs2151280[C]; OR = 1.19, P = 6.9 x 10(-9)), as does rs157935[T] at 7q32 near the imprinted gene KLF14 (OR = 1.23, P = 5.7 x 10(-10)). The effect of rs157935[T] is dependent on the parental origin of the risk allele. None of these variants were found to be associated with melanoma or fair-pigmentation traits. A melanoma- and pigmentation-associated variant in the SLC45A2 gene, L374F, is associated with risk of both BCC and squamous cell carcinoma. Finally, we report conclusive evidence that rs401681[C] in the TERT-CLPTM1L locus confers susceptibility to BCC but protects against melanoma. PMID:19578363

  4. A Complex Interaction Between Reduced Reelin Expression and Prenatal Organophosphate Exposure Alters Neuronal Cell Morphology

    PubMed Central

    Mullen, Brian R.; Ross, Brennan; Chou, Joan Wang; Khankan, Rana; Khialeeva, Elvira; Bui, Kimberly

    2016-01-01

    Genetic and environmental factors are both likely to contribute to neurodevelopmental disorders including schizophrenia, autism spectrum disorders, and major depressive disorders. Prior studies from our laboratory and others have demonstrated that the combinatorial effect of two factors—reduced expression of reelin protein and prenatal exposure to the organophosphate pesticide chlorpyrifos oxon—gives rise to acute biochemical effects and to morphological and behavioral phenotypes in adolescent and young adult mice. In the current study, we examine the consequences of these factors on reelin protein expression and neuronal cell morphology in adult mice. While the cell populations that express reelin in the adult brain appear unchanged in location and distribution, the levels of full length and cleaved reelin protein show persistent reductions following prenatal exposure to chlorpyrifos oxon. Cell positioning and organization in the hippocampus and cerebellum are largely normal in animals with either reduced reelin expression or prenatal exposure to chlorpyrifos oxon, but cellular complexity and dendritic spine organization is altered, with a skewed distribution of immature dendritic spines in adult animals. Paradoxically, combinatorial exposure to both factors appears to generate a rescue of the dendritic spine phenotypes, similar to the mitigation of behavioral and morphological changes observed in our prior study. Together, our observations support an interaction between reelin expression and chlorpyrifos oxon exposure that is not simply additive, suggesting a complex interplay between genetic and environmental factors in regulating brain morphology. PMID:27364165

  5. Circulating Tumor Cells Count and Morphological Features in Breast, Colorectal and Prostate Cancer

    PubMed Central

    Ligthart, Sjoerd T.; Coumans, Frank A. W.; Bidard, Francois-Clement; Simkens, Lieke H. J.; Punt, Cornelis J. A.; de Groot, Marco R.; Attard, Gerhardt; de Bono, Johann S.; Pierga, Jean-Yves; Terstappen, Leon W. M. M.

    2013-01-01

    Background Presence of circulating tumor cells (CTC) in patients with metastatic breast, colorectal and prostate cancer is indicative for poor prognosis. An automated CTC (aCTC) algorithm developed previously to eliminate the variability in manual counting of CTC (mCTC) was used to extract morphological features. Here we validated the aCTC algorithm on CTC images from prostate, breast and colorectal cancer patients and investigated the role of quantitative morphological parameters. Methodology Stored images of samples from patients with prostate, breast and colorectal cancer, healthy controls, benign breast and colorectal tumors were obtained using the CellSearch system. Images were analyzed for the presence of aCTC and their morphological parameters measured and correlated with survival. Results Overall survival hazard ratio was not significantly different for aCTC and mCTC. The number of CTC correlated strongest with survival, whereas CTC size, roundness and apoptosis features reached significance in univariate analysis, but not in multivariate analysis. One aCTC/7.5 ml of blood was found in 7 of 204 healthy controls and 9 of 694 benign tumors. In one patient with benign tumor 2 and another 9 aCTC were detected. Significance of the study CTC can be identified and morphological features extracted by an algorithm on images stored by the CellSearch system and strongly correlate with clinical outcome in metastatic breast, colorectal and prostate cancer. PMID:23826219

  6. Optimization of molecular organization and nanoscale morphology for high performance low bandgap polymer solar cells

    NASA Astrophysics Data System (ADS)

    He, Ming; Wang, Mengye; Lin, Changjian; Lin, Zhiqun

    2014-03-01

    Rational design and synthesis of low bandgap (LBG) polymers with judiciously tailored HOMO and LUMO levels have emerged as a viable route to high performance polymer solar cells with power conversion efficiencies (PCEs) exceeding 10%. In addition to engineering the energy-level of LBG polymers, the photovoltaic performance of LBG polymer-based solar cells also relies on the device architecture, in particular the fine morphology of the photoactive layer. The nanoscale interpenetrating networks composed of nanostructured donor and acceptor phases are the key to providing a large donor-acceptor interfacial area for maximizing the exciton dissociation and offering a continuous pathway for charge transport. In this Review Article, we summarize recent strategies for tuning the molecular organization and nanoscale morphology toward an enhanced photovoltaic performance of LBG polymer-based solar cells.

  7. Morphologic and cytochemical characteristics of blood cells of juvenile loggerhead sea turtles (Caretta caretta).

    PubMed

    Casal, A B; Orós, J

    2007-04-01

    A morphologic classification based on the cytochemical characteristics of blood cells of 35 juvenile loggerhead sea turtles (Caretta caretta) is described. Cytochemical stains included benzidine peroxidase, chloroacetate esterase, alpha-naphthyl butyrate esterase (with and without sodium fluoride), acid phosphatase (with and without tartaric acid), Sudan black B, periodic acid-Schiff, and toluidine blue. The morphologic characteristics of erythrocytes were similar to those reported in green turtles. Six types of white blood cells were identified: heterophils, eosinophils, basophils, lymphocytes, monocytes and thrombocytes. Except for the basophils, the rest of the white blood cells from loggerhead turtles had different cytochemical characteristics compared to blood cells from other sea turtle species. The leukocyte differential count was different from that reported for other sea turtle species. Heterophils were the most numerous leukocytes from these loggerhead turtles, followed by lymphocytes, eosinophils, monocytes and basophils. This paper provides a morphologic classification of blood cells of loggerhead sea turtles that is useful for veterinary surgeons involved in sea turtle conservation.

  8. No Relationship between Embryo Morphology and Successful Derivation of Human Embryonic Stem Cell Lines

    PubMed Central

    Ström, Susanne; Rodriguez-Wallberg, Kenny; Holm, Frida; Bergström, Rosita; Eklund, Linda; Strömberg, Anne-Marie; Hovatta, Outi

    2010-01-01

    Background The large number (30) of permanent human embryonic stem cell (hESC) lines and additional 29 which did not continue growing, in our laboratory at Karolinska Institutet have given us a possibility to analyse the relationship between embryo morphology and the success of derivation of hESC lines. The derivation method has been improved during the period 2002–2009, towards fewer xeno-components. Embryo quality is important as regards the likelihood of pregnancy, but there is little information regarding likelihood of stem cell derivation. Methods We evaluated the relationship of pronuclear zygote stage, the score based on embryo morphology and developmental rate at cleavage state, and the morphology of the blastocyst at the time of donation to stem cell research, to see how they correlated to successful establishment of new hESC lines. Results Derivation of hESC lines succeeded from poor quality and good quality embryos in the same extent. In several blastocysts, no real inner cell mass (ICM) was seen, but permanent well growing hESC lines could be established. One tripronuclear (3PN) zygote, which developed to blastocyst stage, gave origin to a karyotypically normal hESC line. Conclusion Even very poor quality embryos with few cells in the ICM can give origin to hESC lines. PMID:21217828

  9. How well can morphology assess cell death modality? A proteomics study

    PubMed Central

    Chernobrovkin, Alexey L; Zubarev, Roman A

    2016-01-01

    While the focus of attempts to classify cell death programs has finally shifted in 2010s from microscopy-based morphological characteristics to biochemical assays, more recent discoveries have put the underlying assumptions of many such assays under severe stress, mostly because of the limited specificity of the assays. On the other hand, proteomics can quantitatively measure the abundances of thousands of proteins in a single experiment. Thus proteomics could develop a modern alternative to both semiquantitative morphology assessment as well as single-molecule biochemical assays. Here we tested this hypothesis by analyzing the proteomes of cells dying after been treated with various chemical agents. The most striking finding is that, for a multivariate model based on the proteome changes in three cells lines, the regulation patterns of the 200–500 most abundant proteins typically attributed to household type more accurately reflect that of the proteins directly interacting with the drug than any other protein subset grouped by common function or biological process, including cell death. This is in broad agreement with the 'rigid cell death mechanics' model where drug action mechanism and morphological changes caused by it are bijectively linked. This finding, if confirmed, will open way for a broad use of proteomics in death modality assessment. PMID:27752363

  10. Oriented cell division affects the global stress and cell packing geometry of a monolayer under stretch.

    PubMed

    Xu, Guang-Kui; Liu, Yang; Zheng, Zhaoliang

    2016-02-01

    Cell division plays a vital role in tissue morphogenesis and homeostasis, and the division plane is crucial for cell fate. For isolated cells, extensive studies show that the orientation of divisions is sensitive to cell shape and the direction of extrinsic mechanical forces. However, it is poorly understood that how the cell divides within a cell monolayer and how the local stress change, due to the division, affects the global stress of epithelial monolayers. Here, we use the vertex dynamics models to investigate the effects of division orientation on the configurations and mechanics of a cell monolayer under stretch. We examine three scenarios of the divisions: dividing along the stretch axis, dividing along the geometric long axis of cells, and dividing at a random angle. It is found that the division along the long cell axis can induce the minimal energy difference, and the global stress of the monolayer after stretch releases more rapidly in this case. Moreover, the long-axis division can result in more random cell orientations and more isotropic cell shapes within the monolayer, comparing with other two cases. This study helps understand the division orientation of cells within a monolayer under mechanical stimuli, and may shed light on linking individual cell's behaviors to the global mechanics and patterns of tissues. PMID:26774292

  11. Oriented cell division affects the global stress and cell packing geometry of a monolayer under stretch.

    PubMed

    Xu, Guang-Kui; Liu, Yang; Zheng, Zhaoliang

    2016-02-01

    Cell division plays a vital role in tissue morphogenesis and homeostasis, and the division plane is crucial for cell fate. For isolated cells, extensive studies show that the orientation of divisions is sensitive to cell shape and the direction of extrinsic mechanical forces. However, it is poorly understood that how the cell divides within a cell monolayer and how the local stress change, due to the division, affects the global stress of epithelial monolayers. Here, we use the vertex dynamics models to investigate the effects of division orientation on the configurations and mechanics of a cell monolayer under stretch. We examine three scenarios of the divisions: dividing along the stretch axis, dividing along the geometric long axis of cells, and dividing at a random angle. It is found that the division along the long cell axis can induce the minimal energy difference, and the global stress of the monolayer after stretch releases more rapidly in this case. Moreover, the long-axis division can result in more random cell orientations and more isotropic cell shapes within the monolayer, comparing with other two cases. This study helps understand the division orientation of cells within a monolayer under mechanical stimuli, and may shed light on linking individual cell's behaviors to the global mechanics and patterns of tissues.

  12. Morphologic and Gene Expression Criteria for Identifying Human Induced Pluripotent Stem Cells

    PubMed Central

    Wakao, Shohei; Kitada, Masaaki; Kuroda, Yasumasa; Ogura, Fumitaka; Murakami, Toru; Niwa, Akira; Dezawa, Mari

    2012-01-01

    Induced pluripotent stem (iPS) cells can be generated from somatic cells by the forced expression of four factors, Oct3/4, Sox2, Klf4, and c-Myc. While a great variety of colonies grow during induction, only a few of them develop into iPS cells. Researchers currently use visual observation to identify iPS cells and select colonies resembling embryonic stem (ES) cells, and there are no established objective criteria. Therefore, we exhaustively analyzed the morphology and gene expression of all the colonies generated from human fibroblasts after transfection with four retroviral vectors encoding individual factors (192 and 203 colonies in two experiments) and with a single polycistronic retroviral vector encoding all four factors (199 and 192 colonies in two experiments). Here we demonstrate that the morphologic features of emerged colonies can be categorized based on six parameters, and all generated colonies that could be passaged were classified into seven subtypes in colonies transfected with four retroviral vectors and six subtypes with a single polycistronic retroviral vector, both including iPS cell colonies. The essential qualifications for iPS cells were: cells with a single nucleolus; nucleus to nucleolus (N/Nls) ratio ∼2.19: cell size ∼43.5 µm2: a nucleus to cytoplasm (N/C) ratio ∼0.87: cell density in a colony ∼5900 cells/mm2: and number of cell layer single. Most importantly, gene expression analysis revealed for the first time that endogenous Sox2 and Cdx2 were expressed specifically in iPS cells, whereas Oct3/4 and Nanog, popularly used markers for identifying iPS cells, are expressed in colonies other than iPS cells, suggesting that Sox2 and Cdx2 are reliable markers for identifying iPS cells. Our findings indicate that morphologic parameters and the expression of endogenous Sox2 and Cdx2 can be used to accurately identify iPS cells. PMID:23272044

  13. Morphologic and gene expression criteria for identifying human induced pluripotent stem cells.

    PubMed

    Wakao, Shohei; Kitada, Masaaki; Kuroda, Yasumasa; Ogura, Fumitaka; Murakami, Toru; Niwa, Akira; Dezawa, Mari

    2012-01-01

    Induced pluripotent stem (iPS) cells can be generated from somatic cells by the forced expression of four factors, Oct3/4, Sox2, Klf4, and c-Myc. While a great variety of colonies grow during induction, only a few of them develop into iPS cells. Researchers currently use visual observation to identify iPS cells and select colonies resembling embryonic stem (ES) cells, and there are no established objective criteria. Therefore, we exhaustively analyzed the morphology and gene expression of all the colonies generated from human fibroblasts after transfection with four retroviral vectors encoding individual factors (192 and 203 colonies in two experiments) and with a single polycistronic retroviral vector encoding all four factors (199 and 192 colonies in two experiments). Here we demonstrate that the morphologic features of emerged colonies can be categorized based on six parameters, and all generated colonies that could be passaged were classified into seven subtypes in colonies transfected with four retroviral vectors and six subtypes with a single polycistronic retroviral vector, both including iPS cell colonies. The essential qualifications for iPS cells were: cells with a single nucleolus; nucleus to nucleolus (N/Nls) ratio ∼2.19: cell size ∼43.5 µm(2): a nucleus to cytoplasm (N/C) ratio ∼0.87: cell density in a colony ∼5900 cells/mm(2): and number of cell layer single. Most importantly, gene expression analysis revealed for the first time that endogenous Sox2 and Cdx2 were expressed specifically in iPS cells, whereas Oct3/4 and Nanog, popularly used markers for identifying iPS cells, are expressed in colonies other than iPS cells, suggesting that Sox2 and Cdx2 are reliable markers for identifying iPS cells. Our findings indicate that morphologic parameters and the expression of endogenous Sox2 and Cdx2 can be used to accurately identify iPS cells. PMID:23272044

  14. DLC-1, a GTPase-activating protein for Rho, is associated with cell proliferation, morphology, and migration in human hepatocellular carcinoma

    SciTech Connect

    Kim, Tai Young; Lee, Jung Weon; Kim, Hwang-Phill; Jong, Hyun-Soon; Kim, Tae-You; Jung, Mira; Bang, Yung-Jue; E-mail: bangyj@plaza.snu.ac.kr

    2007-03-30

    DLC-1 (deleted in liver cancer-1) is a tumor suppressor gene for hepatocellular carcinoma and other cancers. To characterize its functions, we constructed recombinant adenovirus encoding the wild-type DLC-1 and examined its effects on behaviors of a hepatocellular carcinoma cell line (SNU-368), which does not express DLC-1. Here, we found that restoration of DLC-1 expression in the SNU-368 cells caused an inhibition of cell proliferation with an increase of a subG1 population. Furthermore, DLC-1 overexpression induced disassembly of stress fibers and extensive membrane protrusions around cells on laminin-1. DLC-1 overexpression also inhibited cell migration and dephosphorylated focal adhesion proteins such as focal adhesion kinase (FAK), Cas (p130Cas; Crk-associated substrate), and paxillin. These observations suggest that DLC-1 plays important roles in signal transduction pathway regulating cell proliferation, cell morphology, and cell migration by affecting Rho family GTPases and focal adhesion proteins.

  15. EVALUATION OF BENZO[C]CHRYSENE DIHYDRODIOLS IN THE MORPHOLOGICAL CELL TRANSFORMATION OF MOUSE EMBRYO FIBROBLAST C3H10T1/2CL8 CELLS

    EPA Science Inventory

    EVALUATION OF BENZO[c]CHRYSENE DIHYDRODIOLS IN THE MORPHOLOGICAL CELL TRANSFORMATION OF MOUSE EMBRYO FIBROBLAST C3H10T?CL8 CELLS

    Abstract The morphological cell transforming activities of three dihydrodiols of benzo[c]chrysene (B[c]C), trans-B[c]C-7,8-diol, trans-B[c]C-9...

  16. Anabolic androgens affect the competitive interactions in cell migration and adhesion between normal mouse urothelial cells and urothelial carcinoma cells.

    PubMed

    Huang, Chi-Ping; Hsieh, Teng-Fu; Chen, Chi-Cheng; Hung, Xiao-Fan; Yu, Ai-Lin; Chang, Chawnshang; Shyr, Chih-Rong

    2014-09-26

    The urothelium is constantly rebuilt by normal urothelial cells to regenerate damaged tissues caused by stimuli in urine. However, the urothelial carcinoma cells expand the territory by aberrant growth of tumor cells, which migrate and occupy the damaged tissues to spread outside and disrupt the normal cells and organized tissues and form a tumor. Therefore, the interaction between normal urothelial cells and urothelial carcinoma cells affect the initiation and progression of urothelial tumors if normal urothelial cells fail to migrate and adhere to the damages sites to regenerate the tissues. Here, comparing normal murine urothelial cells with murine urothelial carcinoma cells (MBT-2), we found that normal cells had less migration ability than carcinoma cells. And in our co-culture system we found that carcinoma cells had propensity migrating toward normal urothelial cells and carcinoma cells had more advantages to adhere than normal cells. To reverse this condition, we used anabolic androgen, dihyrotestosterone (DHT) to treat normal cells and found that DHT treatment increased the migration ability of normal urothelial cells toward carcinoma cells and the adhesion capacity in competition with carcinoma cells. This study provides the base of a novel therapeutic approach by using anabolic hormone-enforced normal urothelial cells to regenerate the damage urothelium and defend against the occupancy of carcinoma cells to thwart cancer development and recurrence.

  17. Control over the morphology and segregation of Zebrafish germ cell granules during embryonic development

    PubMed Central

    Strasser, Markus J; Mackenzie, Natalia C; Dumstrei, Karin; Nakkrasae, La-Iad; Stebler, Jürg; Raz, Erez

    2008-01-01

    Background Zebrafish germ cells contain granular-like structures, organized around the cell nucleus. These structures share common features with polar granules in Drosophila, germinal granules in Xenopus and chromatoid bodies in mice germ cells, such as the localization of the zebrafish Vasa, Piwi and Nanos proteins, among others. Little is known about the structure of these granules as well as their segregation in mitosis during early germ-cell development. Results Using transgenic fish expressing a fluorescently labeled novel component of Zebrafish germ cell granules termed Granulito, we followed the morphology and distribution of the granules. We show that whereas these granules initially exhibit a wide size variation, by the end of the first day of development they become a homogeneous population of medium size granules. We investigated this resizing event and demonstrated the role of microtubules and the minus-end microtubule dependent motor protein Dynein in the process. Last, we show that the function of the germ cell granule resident protein the Tudor domain containing protein-7 (Tdrd7) is required for determination of granule morphology and number. Conclusion Our results suggest that Zebrafish germ cell granules undergo a transformation process, which involves germ cell specific proteins as well as the microtubular network. PMID:18507824

  18. Clinicopathologic study on combined hepatocellular carcinoma and cholangiocarcinoma: with emphasis on the intermediate cell morphology.

    PubMed

    Park, Ho Sung; Bae, Jun Sang; Jang, Kyu Yun; Lee, Ju Hyung; Yu, Hee Chul; Jung, Ji Hyeon; Cho, Baik Hwan; Chung, Myoung Ja; Moon, Woo Sung

    2011-08-01

    Combined hepatocellular carcinoma and cholangiocarcinoma (combined HCC-CC) is a rare subtype of primary liver cancer. We investigated the histopathologic features of transitional or intermediate areas in 21 combined HCC-CCs and immunophenotypes using different hepatic progenitor cell markers (CK7, CK19, c-kit, NCAM, and EpCAM). Major histologic findings of transitional or intermediate areas of 21 combined HCC-CCs included strands/trabeculae of small, uniform, oval-shaped cells with scant cytoplasm and hyperchromatic nuclei embedded within an abundant stroma, small cells with an antler-like anastomosing pattern, and solid nests of intermediate hepatocyte-like cells surrounded by small cells in periphery, in order of frequency. The intermediate area of one tumor was composed predominantly of spindle cells arranged in short fascicles. Immunophenotype of tumor cells with intermediate morphology suggested a progenitor cell origin for this tumor. Clinical findings of combined HCC-CC showed a closer resemblance with those of HCC than those of CC. In univariate analysis, tumor size, TNM stage, and serum alpha-fetoprotein levels showed a significant association with poor patient survival. Serum alpha-fetoprotein level was an independent prognostic indicator in multivariate analysis. In conclusion, an awareness of the clinicopathologic features, specifically the various morphologic features of intermediate areas in this tumor, is essential for prevention of potential misdiagnosis as another tumor.

  19. Solitary cutaneous histiocytosis with granular cell changes: a morphological variant of reticulohistiocytoma?

    PubMed

    Caltabiano, Rosario; Magro, Gaetano; Vecchio, Giada Maria; Lanzafame, Salvatore

    2010-02-01

    We first report a case of granular cell histiocytosis occurring as a solitary polypoid lesion of the nipple in a 15-year-old girl. Histologically, the lesion was composed of a dermal population of medium- to large-sized, short spindle- to round- to epithelioid-shaped cells with eosinophilic cytoplasm containing numerous and small diastase-resistant periodic acid-Schiff (PAS) positive granules. No associated inflammatory cells were observed. Immunohistochemical studies, revealing immunoreactivity exclusively to vimentin and CD68, were consistent with their histiocytic profile. Based on clinical, morphological and immunohistochemical features, the diagnosis of 'solitary cutaneous histiocytosis with granular cell changes' was proposed. The absence of an inflammatory cell component, such as lymphocytes and leucocytes, along with no history of a previous trauma or medical treatment, suggest that the present lesion could fit into the morphological spectrum of the so-called solitary epithelioid histiocytoma, also known as reticulohistiocytoma. Alternatively, the possibility of a histiocytic reaction to unknown stimuli cannot be completely ruled out. Nevertheless, awareness of solitary cutaneous histiocytosis with granular cell changes is useful to avoid confusion with other dermal tumors, especially 'granular cell tumor' and 'dermal non-neural granular cell tumor'.

  20. Putative role of border cells in generating spontaneous morphological activity within Kölliker's organ.

    PubMed

    Dayaratne, M W Nishani; Vlajkovic, Srdjan M; Lipski, Janusz; Thorne, Peter R

    2015-12-01

    Kölliker's organ is a transient epithelial structure, comprising a major part of the organ of Corti during pre-hearing stages of development. The auditory system is spontaneously active during development, which serves to retain and refine neural connections. Kölliker's organ is considered a key candidate for generating such spontaneous activity, most likely through purinergic (P2 receptor) signalling and inner hair cell (IHC) activation. Associated with the spontaneous neural activity, ATP released locally by epithelial cells induces rhythmic morphological changes within Kölliker's organ, the purpose of which is not understood. These changes are accompanied by a shift in cellular refractive index, allowing optical detection of this activity in real-time. Using this principle, we investigated the origin of spontaneous morphological activity within Kölliker's organ. Apical turns of Wistar rat cochleae (P9-11) were dissected, and the purinergic involvement was studied following acute tissue exposure to a P2 receptor agonist (ATPγS) and antagonist (suramin). ATPγS induced a sustained darkening throughout Kölliker's organ, reversed by suramin. This effect was most pronounced in the region closest to the inner hair cells, which also displayed the highest frequency of intrinsic morphological events. Additionally, suramin alone induced swelling of this region, suggesting a tight regulation of cell volume by ATP-mediated mechanisms. Histological analysis of cochlear tissues demonstrates the most profound volume changes in the border cell region immediately adjacent to the IHCs. Together, these results underline the role of purinergic signalling in initiating morphological events within Kölliker's organ, and suggest a key involvement of border cells surrounding IHCs in regulating this spontaneous activity.

  1. Overexpression of a grapevine R2R3-MYB factor in tomato affects vegetative development, flower morphology and flavonoid and terpenoid metabolism.

    PubMed

    Mahjoub, Ali; Hernould, Michel; Joubès, Jérôme; Decendit, Alain; Mars, Mohamed; Barrieu, François; Hamdi, Saïd; Delrot, Serge

    2009-07-01

    Although the terpenoid pathway constitutes, with the phenylpropanoid metabolism, the major pathway of secondary metabolism in plants, little is known about its regulation. Overexpression of a Vitis vinifera R2R3-MYB transcription factor (VvMYB5b) in tomato induced pleiotropic changes including dwarfism, modified leaf structure, alterations of floral morphology, pigmented and glossy fruits at the "green-mature" stage and impaired seed germination. Two main branches of secondary metabolism, which profoundly influence the organoleptic properties of the fruit, were affected in the opposite way by VvMYB5b overexpression. Phenylpropanoid metabolism was down regulated whereas the amount of beta-carotene was up regulated. This is the first example of the independent regulation of phenylpropanoid and carotenoid metabolism. The strongest modification concerns a decrease in beta-amyrin, the precursor of the oleanolic acid, which is the major component of grape waxes. Scanning electron microscopy analysis of fruits and leaves confirms the alteration of wax metabolism and a modification of cell size and shape. This may potentially impact resistance/tolerance to biotic and abiotic stresses. The results are compared with a similar approach using heterologous expression of VvMYB5b in tobacco. PMID:19375343

  2. Aluminium oxide nanoparticles induced morphological changes, cytotoxicity and oxidative stress in Chinook salmon (CHSE-214) cells.

    PubMed

    Srikanth, Koigoora; Mahajan, Amit; Pereira, Eduarda; Duarte, Armando Costa; Venkateswara Rao, Janapala

    2015-10-01

    Aluminium oxide nanoparticles (Al2 O3 NPs) are increasingly used in diverse applications that has raised concern about their safety. Recent studies suggested that Al2 O3 NPs induced oxidative stress may be the cause of toxicity in algae, Ceriodaphnia dubia, Caenorhabditis elegans and Danio rerio. However, there is paucity on the toxicity of Al2 O3 NPs on fish cell lines. The current study was aimed to investigate Al2 O3 NPs induced cytotoxicity, oxidative stress and morphological abnormality of Chinnok salmon cells (CHSE-214). A dose-dependent decline in cell viability was observed in CHSE-214 cells exposed to Al2 O3 NPs. Oxidative stress induced by Al2 O3 NPs in CHSE-214 cells has resulted in the significant reduction of superoxide dismutase, catalase and glutathione in a dose-dependent manner. However, a significant increase in glutathione sulfo-transferase and lipid peroxidation was observed in CHSE-214 cells exposed to Al2 O3 NPs in a dose-dependent manner. Significant morphological changes in CHSE-214 cells were observed when exposed to Al2 O3 NPs at 6, 12 and 24 h. The cells started to detach and appear spherical at 6 h followed by loss of cellular contents resulting in the shrinking of the cells. At 24 h, the cells started to disintegrate and resulted in cell death. Our data demonstrate that Al2 O3 NPs induce cytotoxicity and oxidative stress in a dose-dependent manner in CHSE-214 cells. Thus, our current work may serve as a base-line study for future evaluation of toxicity studies using CHSE-214 cells. PMID:25875951

  3. Investigation on 3D morphological changes of in vitro cells through digital holographic microscopy

    NASA Astrophysics Data System (ADS)

    Memmolo, Pasquale; Miccio, Lisa; Merola, Francesco; Netti, Paolo A.; Coppola, Giuseppe; Ferraro, Pietro

    2013-04-01

    We report the investigation of the identification and measurement of region of interest (ROI) in quantitative phase-contrast maps (QPMs) of biological cells by digital holographic microscopy (DHM), with the aim to analyze the 3D positions and 3D morphology together. We consider as test case for our tool the in vitro bull sperm head morphometry analysis. Extraction and measurement of various morphological parameters are performed by using two methods: the anisotropic diffusion filter, that is based on the Gaussian diffusivity function which allows more accuracy of the edge position, and the simple thresholding filter. In particular we consider the calculation of area, ellipticity, perimeter, major axis, minor axis and shape factor as a morphological parameter, instead, for the estimation of 3D position, we compute the centroid, the weighted centroid and the maximum phase values. A statistical analysis on a data set composed by N = 14 holograms relative to bovine spermatozoa and its reference holograms is reported.

  4. Immunohistowax processing, a new fixation and embedding method for light microscopy, which preserves antigen immunoreactivity and morphological structures: visualisation of dendritic cells in peripheral organs

    PubMed Central

    Pajak, B.; De Smedt, T.; Moulin, V.; De Trez, C.; Maldonado-Lopez, R.; Vansanten, G.; Briend, E.; Urbain, J.; Leo, O.; Moser, M.

    2000-01-01

    Aims—To describe a new fixation and embedding method for tissue samples, immunohistowax processing, which preserves both morphology and antigen immunoreactivity, and to use this technique to investigate the role of dendritic cells in the immune response in peripheral tissues. Methods—This technique was used to stain a population of specialised antigen presenting cells (dendritic cells) that have the unique capacity to sensitise naive T cells, and therefore to induce primary immune responses. The numbers of dendritic cells in peripheral organs of mice either untreated or injected with live Escherichia coli were compared. Results—Numbers of dendritic cells were greatly decreased in heart, kidney, and intestine after the inoculation of bacteria. The numbers of dendritic cells in the lung did not seem to be affected by the injection of E coli. However, staining of lung sections revealed that some monocyte like cells acquired morphological and phenotypic features of dendritic cells, and migrated into blood vessels. Conclusions—These observations suggest that the injection of bacteria induces the activation of dendritic cells in peripheral organs, where they play the role of sentinels, and/or their movement into lymphoid organs, where T cell priming is likely to occur. Key Words: dendritic cell • Escherichia coli • immunohistochemistry PMID:10961175

  5. Block copolymer morphologies in dye-sensitized solar cells: probing the photovoltaic structure-function relation.

    PubMed

    Crossland, Edward J W; Nedelcu, Mihaela; Ducati, Caterina; Ludwigs, Sabine; Hillmyer, Marc A; Steiner, Ullrich; Snaith, Henry J

    2009-08-01

    We integrate mesostructured titania arrays into dye-sensitized solar cells by replicating ordered, oriented one-dimensional (1D) columnar and three-dimensional (3D) bicontinuous gyroid block copolymer phases. The solar cell performance, charge transport, and recombination are investigated. We observe faster charge transport in 1D "wires" than through 3D gyroid arrays. However, owing to their structural instability, the surface area of the wire arrays is low, inhibiting the solar cell performance. The gyroid morphology, on the other hand, outperforms the current state-of-the-art mesoporous nanoparticle films.

  6. The selective role of ECM components on cell adhesion, morphology, proliferation and communication in vitro

    SciTech Connect

    Schlie-Wolter, Sabrina; Ngezahayo, Anaclet; Chichkov, Boris N.

    2013-06-10

    Cell binding to the extracellular matrix (ECM) is essential for cell and tissue functions. In this context, each tissue consists of a unique ECM composition, which may be responsible for tissue-specific cell responses. Due to the complexity of ECM-cell interactions—which depend on the interplay of inside-out and outside-in signaling cascades, cell and tissue specificity of ECM-guidance is poorly understood. In this paper, we investigate the role of different ECM components like laminin, fibronectin, and collagen type I with respect to the essential cell behaviour patterns: attachment dynamics such as adhesion kinetic and force, formation of focal adhesion complexes, morphology, proliferation, and intercellular communication. A detailed in vitro comparison of fibroblasts, endothelial cells, osteoblasts, smooth muscle cells, and chondrocytes reveals significant differences in their cell responses to the ECM: cell behaviour follows a cell specific ligand priority ranking, which was independent of the cell type origin. Fibroblasts responded best to fibronectin, chondrocytes best to collagen I, the other cell types best to laminin. This knowledge is essential for optimization of tissue-biomaterial interfaces in all tissue engineering applications and gives insight into tissue-specific cell guidance. -- Highlights: • We analyse the impact of ECM components on cell behaviour in vitro. • We compare five different cell types, using the same culture conditions. • The ECM significantly guides all cell responses. • Cell behaviour follows a cell specific ligand-priority ranking. • This gives insight in tissue formation and is essential for biomedical applications.

  7. Effects of Laser Peripheral Iridotomy on Corneal Endothelial Cell Density and Cell Morphology in Primary Angle Closure Suspect Subjects

    PubMed Central

    Jamali, Hossein; Jahanian, Sara; Gharebaghi, Reza

    2016-01-01

    Purpose: To evaluate the effects of prophylactic laser peripheral iridotomy on corneal endothelial cell density and cell morphology in subjects with primary angle closure suspect (PACS) within a one-year follow-up period. Methods: In this quasi-experimental prospective study, from June 2012 to November 2013, thirty-five PACS eyes underwent laser peripheral iridotomy at clinics affiliated to Shiraz University of Medical Sciences, Shiraz, Iran. After obtaining informed consent, specular microscopy was performed at baseline and at 3-month, 6-month and 12-month follow-up visits. Central, nasal and temporal endothelial cell counts and cell morphology were evaluated via non-contact specular microscopy. Results: The mean subject age was 53.4 ± 7.9 years, and the majority of subjects were women (88.2%). The mean central corneal endothelial cell count prior to laser peripheral iridotomy was 2528 ± 119.2, and this value changed to 2470 ± 175.9, 2425 ± 150.6, and 2407 ± 69.02 at the 3-month, 6-month, and 12-month follow-up visits, respectively; these differences did not reach statistical significance. Additionally, the changes in the number of cells, the hexagonality of cells, and the coefficient of variation (CV) in the central, nasal, and temporal areas were not significant. Conclusion: In PACS eyes, we did not find a decline in corneal endothelial cell density or a change in cell morphological characteristics, including cell hexagonality and CV, in the central, nasal, and temporal regions of the cornea in any of our subjects over a one-year follow-up period. PMID:27621781

  8. Effects of Laser Peripheral Iridotomy on Corneal Endothelial Cell Density and Cell Morphology in Primary Angle Closure Suspect Subjects

    PubMed Central

    Jamali, Hossein; Jahanian, Sara; Gharebaghi, Reza

    2016-01-01

    Purpose: To evaluate the effects of prophylactic laser peripheral iridotomy on corneal endothelial cell density and cell morphology in subjects with primary angle closure suspect (PACS) within a one-year follow-up period. Methods: In this quasi-experimental prospective study, from June 2012 to November 2013, thirty-five PACS eyes underwent laser peripheral iridotomy at clinics affiliated to Shiraz University of Medical Sciences, Shiraz, Iran. After obtaining informed consent, specular microscopy was performed at baseline and at 3-month, 6-month and 12-month follow-up visits. Central, nasal and temporal endothelial cell counts and cell morphology were evaluated via non-contact specular microscopy. Results: The mean subject age was 53.4 ± 7.9 years, and the majority of subjects were women (88.2%). The mean central corneal endothelial cell count prior to laser peripheral iridotomy was 2528 ± 119.2, and this value changed to 2470 ± 175.9, 2425 ± 150.6, and 2407 ± 69.02 at the 3-month, 6-month, and 12-month follow-up visits, respectively; these differences did not reach statistical significance. Additionally, the changes in the number of cells, the hexagonality of cells, and the coefficient of variation (CV) in the central, nasal, and temporal areas were not significant. Conclusion: In PACS eyes, we did not find a decline in corneal endothelial cell density or a change in cell morphological characteristics, including cell hexagonality and CV, in the central, nasal, and temporal regions of the cornea in any of our subjects over a one-year follow-up period.

  9. Endosulfan affects GnRH cells in sexually differentiated juveniles of the perciform Cichlasoma dimerus.

    PubMed

    Piazza, Yanina; Pandolfi, Matías; Da Cuña, Rodrigo; Genovese, Griselda; Lo Nostro, Fabiana

    2015-06-01

    Endosulfan (ES) is an organochlorine pesticide widely used in agriculture despite its high toxicity towards non-target organisms such as fish. It has been demonstrated that ES can cause negative effects on aquatic animals, including disruption of hormonal systems. However, the alterations produced by this pesticide on the reproductive axis of fish prior to sexual maturity, as well as possible modes of action have hardly been studied. This study aimed at assessing the effect of waterborne exposure to the pesticide ES on the reproductive axis during sexual differentiation of juveniles of the South American freshwater cichlid fish Cichlasoma dimerus. No mortality was observed due to ES subchronic exposure (90 days post-fertilization). Exposure to ES did not affect body weight nor morphometric parameters, indicating that larvae nutritional state was not affected. Timing of sexual differentiation, gonadal morphology and sex ratio were likewise not altered by ES. However, ES acted as an endocrine disrupting chemical in this species as the morphometry of gonadotropin-releasing hormones (GnRH) producing cells was altered. Exposure to ES altered nuclear area, cell area and nucleus/cytoplasm ratio of GnRH II neurons, and cell and nuclear area and diameter of GnRH III neurons. Interestingly, in our previous study, exposure before sex differentiation (30 day exposure) caused no alteration to GnRH II and III, and did alter GnRH I and FSH cells. These alterations could lead to changes in circulating hormone levels, especially when fish are exposed for prolonged periods, ultimately impairing reproductive fitness. C. dimerus juveniles can be an interesting biological model to perform toxicological studies with the intent to assess early disruption endpoints in the reproductive axis during development. PMID:25800987

  10. Endosulfan affects GnRH cells in sexually differentiated juveniles of the perciform Cichlasoma dimerus.

    PubMed

    Piazza, Yanina; Pandolfi, Matías; Da Cuña, Rodrigo; Genovese, Griselda; Lo Nostro, Fabiana

    2015-06-01

    Endosulfan (ES) is an organochlorine pesticide widely used in agriculture despite its high toxicity towards non-target organisms such as fish. It has been demonstrated that ES can cause negative effects on aquatic animals, including disruption of hormonal systems. However, the alterations produced by this pesticide on the reproductive axis of fish prior to sexual maturity, as well as possible modes of action have hardly been studied. This study aimed at assessing the effect of waterborne exposure to the pesticide ES on the reproductive axis during sexual differentiation of juveniles of the South American freshwater cichlid fish Cichlasoma dimerus. No mortality was observed due to ES subchronic exposure (90 days post-fertilization). Exposure to ES did not affect body weight nor morphometric parameters, indicating that larvae nutritional state was not affected. Timing of sexual differentiation, gonadal morphology and sex ratio were likewise not altered by ES. However, ES acted as an endocrine disrupting chemical in this species as the morphometry of gonadotropin-releasing hormones (GnRH) producing cells was altered. Exposure to ES altered nuclear area, cell area and nucleus/cytoplasm ratio of GnRH II neurons, and cell and nuclear area and diameter of GnRH III neurons. Interestingly, in our previous study, exposure before sex differentiation (30 day exposure) caused no alteration to GnRH II and III, and did alter GnRH I and FSH cells. These alterations could lead to changes in circulating hormone levels, especially when fish are exposed for prolonged periods, ultimately impairing reproductive fitness. C. dimerus juveniles can be an interesting biological model to perform toxicological studies with the intent to assess early disruption endpoints in the reproductive axis during development.

  11. Distinct Morphology of Human T-Cell Leukemia Virus Type 1-Like Particles.

    PubMed

    Maldonado, José O; Cao, Sheng; Zhang, Wei; Mansky, Louis M

    2016-01-01

    The Gag polyprotein is the main retroviral structural protein and is essential for the assembly and release of virus particles. In this study, we have analyzed the morphology and Gag stoichiometry of human T-cell leukemia virus type 1 (HTLV-1)-like particles and authentic, mature HTLV-1 particles by using cryogenic transmission electron microscopy (cryo-TEM) and scanning transmission electron microscopy (STEM). HTLV-1-like particles mimicked the morphology of immature authentic HTLV-1 virions. Importantly, we have observed for the first time that the morphology of these virus-like particles (VLPs) has the unique local feature of a flat Gag lattice that does not follow the curvature of the viral membrane, resulting in an enlarged distance between the Gag lattice and the viral membrane. Other morphological features that have been previously observed with other retroviruses include: (1) a Gag lattice with multiple discontinuities; (2) membrane regions associated with the Gag lattice that exhibited a string of bead-like densities at the inner leaflet; and (3) an arrangement of the Gag lattice resembling a railroad track. Measurement of the average size and mass of VLPs and authentic HTLV-1 particles suggested a consistent range of size and Gag copy numbers in these two groups of particles. The unique local flat Gag lattice morphological feature observed suggests that HTLV-1 Gag could be arranged in a lattice structure that is distinct from that of other retroviruses characterized to date. PMID:27187442

  12. Distinct Morphology of Human T-Cell Leukemia Virus Type 1-Like Particles

    PubMed Central

    Maldonado, José O.; Cao, Sheng; Zhang, Wei; Mansky, Louis M.

    2016-01-01

    The Gag polyprotein is the main retroviral structural protein and is essential for the assembly and release of virus particles. In this study, we have analyzed the morphology and Gag stoichiometry of human T-cell leukemia virus type 1 (HTLV-1)-like particles and authentic, mature HTLV-1 particles by using cryogenic transmission electron microscopy (cryo-TEM) and scanning transmission electron microscopy (STEM). HTLV-1-like particles mimicked the morphology of immature authentic HTLV-1 virions. Importantly, we have observed for the first time that the morphology of these virus-like particles (VLPs) has the unique local feature of a flat Gag lattice that does not follow the curvature of the viral membrane, resulting in an enlarged distance between the Gag lattice and the viral membrane. Other morphological features that have been previously observed with other retroviruses include: (1) a Gag lattice with multiple discontinuities; (2) membrane regions associated with the Gag lattice that exhibited a string of bead-like densities at the inner leaflet; and (3) an arrangement of the Gag lattice resembling a railroad track. Measurement of the average size and mass of VLPs and authentic HTLV-1 particles suggested a consistent range of size and Gag copy numbers in these two groups of particles. The unique local flat Gag lattice morphological feature observed suggests that HTLV-1 Gag could be arranged in a lattice structure that is distinct from that of other retroviruses characterized to date. PMID:27187442

  13. Observation of dendritic cell morphology under light, phase-contrast or confocal laser scanning microscopy.

    PubMed

    Tan, Yuen-Fen; Leong, Chooi-Fun; Cheong, Soon-Keng

    2010-12-01

    Dendritic cells (DCs) are professional antigen presenting cells of the immune system. They can be generated in vitro from peripheral blood monocytes supplemented with GM-CSF, IL-4 and TNF alpha. During induction, DCs will increase in size and acquire multiple cytoplasmic projections when compared to their precursor cells such as monocytes or haematopoietic stem cells which are usually round or spherical. Morphology of DCs can be visualized by conventional light microscopy after staining or phase-contrast inverted microscopy or confocal laser scanning microscopy. In this report, we described the morphological appearances of DCs captured using the above-mentioned techniques. We found that confocal laser scanning microscopy yielded DCs images with greater details but the operating cost for such a technique is high. On the other hand, the images obtained through light microscopy after appropriate staining or phase contrast microscopy were acceptable for identification purpose. Besides, these equipments are readily available in most laboratories and the cost of operation is affordable. Nevertheless, morphological identification is just one of the methods to characterise DCs. Other methods such as phenotypic expression markers and mixed leukocyte reactions are additional tools used in the characterisation of DCs. PMID:21329180

  14. The morphologies of breast cancer cell lines in three-dimensionalassays correlate with their profiles of gene expression

    SciTech Connect

    Kenny, Paraic A.; Lee, Genee Y.; Myers, Connie A.; Neve, RichardM.; Semeiks, Jeremy R.; Spellman, Paul T.; Lorenz, Katrin; Lee, Eva H.; Barcellos-Hoff, Mary Helen; Petersen, Ole W.; Gray, Joe W.; Bissell, MinaJ.

    2007-01-31

    3D cell cultures are rapidly becoming the method of choice for the physiologically relevant modeling of many aspects of non-malignant and malignant cell behavior ex vivo. Nevertheless, only a limited number of distinct cell types have been evaluated in this assay to date. Here we report the first large scale comparison of the transcriptional profiles and 3D cell culture phenotypes of a substantial panel of human breast cancer cell lines. Each cell line adopts a colony morphology of one of four main classes in 3D culture. These morphologies reflect, at least in part, the underlying gene expression profile and protein expression patterns of the cell lines, and distinct morphologies were also associated with tumor cell invasiveness and with cell lines originating from metastases. We further demonstrate that consistent differences in genes encoding signal transduction proteins emerge when even tumor cells are cultured in 3D microenvironments.

  15. Factors affecting the cryosurvival of mouse two-cell embryos.

    PubMed

    Critser, J K; Arneson, B W; Aaker, D V; Huse-Benda, A R; Ball, G D

    1988-01-01

    A series of 4 experiments was conducted to examine factors affecting the survival of frozen-thawed 2-cell mouse embryos. Rapid addition of 1.5 M-DMSO (20 min equilibration at 25 degrees C) and immediate, rapid removal using 0.5 M-sucrose did not alter the frequency (mean +/- s.e.m.) of blastocyst development in vitro when compared to untreated controls (90.5 +/- 2.7% vs 95.3 +/- 2.8%). There was an interaction between the temperature at which slow cooling was terminated and thawing rate. Termination of slow cooling (-0.3 degrees C/min) at -40 degrees C with subsequent rapid thawing (approximately 1500 degrees C/min) resulted in a lower frequency of blastocyst development than did termination of slow cooling at -80 degrees C with subsequent slow thawing (+8 degrees C/min) (36.8 +/- 5.6% vs 63.9 +/- 5.7%). When slow cooling was terminated between -40 and -60 degrees C, higher survival rates were achieved with rapid thawing. When slow cooling was terminated below -60 degrees C, higher survival rates were obtained with slow thawing rates. In these comparisons absolute survival rates were highest among embryos cooled below -60 degrees C and thawed slowly. However, when slow cooling was terminated at -32 degrees C, with subsequent rapid warming, survival rates were not different from those obtained when embryos were cooled to -80 degrees C and thawed slowly (52.4 +/- 9.5%, 59.5 +/- 8.6%). These results suggest that optimal cryosurvival rates may be obtained from 2-cell mouse embryos by a rapid or slow thawing procedure, as has been found for mouse preimplantation embryos at later stages.(ABSTRACT TRUNCATED AT 250 WORDS)

  16. Cations in mammalian cells and chromosomes: Sample preparation protocols affect elemental abundances by SIMS

    NASA Astrophysics Data System (ADS)

    Levi-Setti, R.; Gavrilov, K. L.; Neilly, M. E.

    2006-07-01

    The focus of our current research aims at detailing and quantifying the presence of cations, primarily Ca and Mg, in mammalian cells and chromosomes throughout the different stages of the cell cycle, using our high resolution scanning ion microprobe, the UC-SIM. The 45 keV Ga + probe of this instrument, typically ˜40 nm in diameter, carries a current of 30-40 pA, appropriate for surface SIMS studies, but limited in sample erosion rate for dynamic SIMS mapping over cell-size areas, of order 100 μm × 100 μm. Practical and reliable use of this probe toward the above SIMS goals requires a careful matching of the latter factors with the physical and chemical consequences of sample preparation protocols. We examine here how the preferred sample cryo-preservation methodologies such as freeze-fracture and lyophilization affect high resolution SIMS analysis, and, from this standpoint, develop and evaluate the advantages and disadvantages of fast alternate approaches to drying frozen samples. The latter include the use of methanol, ethanol, and methanol/acetic acid fixative. Methanol-dried freeze-fractured samples preserve histological morphology and yield Ca and Mg distributions containing reliable differential dynamical information, when compared with those following lyophilization.

  17. Enteropathogenic and enterohemorrhagic Escherichia coli type III secretion effector EspV induces radical morphological changes in eukaryotic cells.

    PubMed

    Arbeloa, Ana; Oates, Clare V; Marchès, Oliver; Hartland, Elizabeth L; Frankel, Gad

    2011-03-01

    Enteropathogenic Escherichia coli (EPEC) and enterohemorrhagic Escherichia coli (EHEC) are important human pathogens that rely on translocation of type III secretion system (T3SS) effectors for subversion of signal transduction pathways and colonization of the mammalian gut mucosa. While a core set of effectors is conserved between EPEC and EHEC strains, a growing number of accessory effectors that were found at various frequencies in clinical and environmental isolates have been recently identified. Recent genome projects identified espV as a pseudogene in EHEC but a putative functional gene in EPEC strains E110019 and E22 and the closely related mouse pathogen Citrobacter rodentium. The aim of this study was to determine the distribution of espV among clinical EPEC and EHEC strains and to investigate its function and role in pathogenesis. espV was found in 16% of the tested strains. While deletion of espV from C. rodentium did not affect colonization dynamics or fitness in mixed infections, expression of EspV in mammalian cells led to drastic morphological alterations, which were characterized by nuclear condensation, cell rounding, and formation of dendrite-like projections. Expression of EspV in yeast resulted in a dramatic increase in cell size and irreversible growth arrest. Although the role of EspV in infection and its target host cell protein(s) require further investigation, the data point to a novel mechanism by which the T3SS subverts cell signaling.

  18. Stiffness of hyaluronic acid gels containing liver extracellular matrix supports human hepatocyte function and alters cell morphology.

    PubMed

    Deegan, Daniel B; Zimmerman, Cynthia; Skardal, Aleksander; Atala, Anthony; Shupe, Thomas D

    2015-03-01

    Tissue engineering and cell based liver therapies have utilized primary hepatocytes with limited success due to the failure of hepatocytes to maintain their phenotype in vitro. In order to overcome this challenge, hyaluronic acid (HA) cell culture substrates were formulated to closely mimic the composition and stiffness of the normal liver cellular microenvironment. The stiffness of the substrate was modulated by adjusting HA hydrogel crosslinking. Additionally, the repertoire of bioactive molecules within the HA substrate was bolstered by supplementation with normal liver extracellular matrix (ECM). Primary human hepatocyte viability and phenotype were determined over a narrow physiologically relevant range of substrate stiffnesses from 600 to 4600Pa in both the presence and absence of liver ECM. Cell attachment, viability, and organization of the actin cytoskeleton improved with increased stiffness up to 4600Pa. These differences were not evident in earlier time points or substrates containing only HA. However, gene expression for the hepatocyte markers hepatocyte nuclear factor 4 alpha (HNF4α) and albumin significantly decreased on the 4600Pa stiffness at day 7 indicating that cells may not have maintained their phenotype long-term at this stiffness. Function, as measured by albumin secretion, varied with both stiffness and time in culture and peaked at day 7 at the 1200Pa stiffness, slightly below the stiffness of normal liver ECM at 3000Pa. Overall, gel stiffness affected primary human hepatocyte cell adhesion, functional marker expression, and morphological characteristics dependent on both the presence of liver ECM in gel substrates and time in culture.

  19. Stiffness of hyaluronic acid gels containing liver extracellular matrix supports human hepatocyte function and alters cell morphology.

    PubMed

    Deegan, Daniel B; Zimmerman, Cynthia; Skardal, Aleksander; Atala, Anthony; Shupe, Thomas D

    2015-03-01

    Tissue engineering and cell based liver therapies have utilized primary hepatocytes with limited success due to the failure of hepatocytes to maintain their phenotype in vitro. In order to overcome this challenge, hyaluronic acid (HA) cell culture substrates were formulated to closely mimic the composition and stiffness of the normal liver cellular microenvironment. The stiffness of the substrate was modulated by adjusting HA hydrogel crosslinking. Additionally, the repertoire of bioactive molecules within the HA substrate was bolstered by supplementation with normal liver extracellular matrix (ECM). Primary human hepatocyte viability and phenotype were determined over a narrow physiologically relevant range of substrate stiffnesses from 600 to 4600Pa in both the presence and absence of liver ECM. Cell attachment, viability, and organization of the actin cytoskeleton improved with increased stiffness up to 4600Pa. These differences were not evident in earlier time points or substrates containing only HA. However, gene expression for the hepatocyte markers hepatocyte nuclear factor 4 alpha (HNF4α) and albumin significantly decreased on the 4600Pa stiffness at day 7 indicating that cells may not have maintained their phenotype long-term at this stiffness. Function, as measured by albumin secretion, varied with both stiffness and time in culture and peaked at day 7 at the 1200Pa stiffness, slightly below the stiffness of normal liver ECM at 3000Pa. Overall, gel stiffness affected primary human hepatocyte cell adhesion, functional marker expression, and morphological characteristics dependent on both the presence of liver ECM in gel substrates and time in culture. PMID:26569044

  20. Influence of carvacrol and 1,8-cineole on cell viability, membrane integrity, and morphology of Aeromonas hydrophila cultivated in a vegetable-based broth.

    PubMed

    de Sousa, Jossana Pereira; de Oliveira, Kataryne Árabe Rimá; de Figueiredo, Regina Celia Bressan Queiroz; de Souza, Evandro Leite

    2015-02-01

    This study investigated the effects of carvacrol (CAR) and 1,8-cineole (CIN) alone (at the MIC) or in combination at subinhibitory amounts (both at 1/8 MIC) on the cell viability, membrane permeability, and morphology of Aeromonas hydrophila INCQS 7966 (A. hydrophila) cultivated in a vegetable-based broth. CAR and CIN alone or in combination severely affected the viability of the bacteria and caused dramatic changes in the cell membrane permeability, leading to cell death, as observed by confocal laser microscopy. Scanning and transmission electron microscopy images of bacterial cells exposed to CAR or CIN or the mixture of both compounds revealed severe changes in cell wall structure, rupture of the plasma membrane, shrinking of cells, condensation of cytoplasmic content, leakage of intracellular material, and cell collapse. These findings suggest that CAR and CIN alone or in combination at subinhibitory amounts could be applied to inhibit the growth of A. hydrophila in foods, particularly as sanitizing agents in vegetables.

  1. Controllable synthesis of ZnO nanograss with different morphologies and enhanced performance in dye-sensitized solar cells

    SciTech Connect

    Zhu Shibu; Chen Xiangnan; Zuo Feibiao; Jiang Man; Zhou Zuowan; Hui, David

    2013-01-15

    A series of ZnO nanograss films grown on fluorine-doped tin oxide coated glass substrates were synthesized via hydrothermal method by using polyethyleneimine (PEI) as adjusting agent. The films were characterized by field emission scanning electron microscope (FE-SEM) and X-ray diffraction (XRD). It was found that the PEI not only affected the aspect ratios of ZnO nanograss but also changed the geometrical shape of ZnO nanograss. A possible mechanism based on PEI adsorbed on the non-polar facets of ZnO that governed the growth rate of different directions were proposed to elucidate the effect of PEI on morphology of ZnO. The ZnO nanograss films were applied to dye-sensitized solar cells (DSSCs). The results showed that the photocurrent density significantly enhanced, and the power conversion efficiency increased by 55% based on ZnO nanograss synthesized in a growth solution containing 7 mmol/L PEI, resulting from the dye loading properties related to the different morphologies. - Graphical abstract: Effect of PEI on ZnO nanograss: controlling the aspect ratio and morphology of ZnO and enhancing their photovoltaic performance. Highlights: Black-Right-Pointing-Pointer ZnO nanograss with different aspect ratios were synthesized by adjusting PEI content. Black-Right-Pointing-Pointer PEI affects both on the aspect ratios and geometrical shapes of ZnO nanograss. Black-Right-Pointing-Pointer ZnO nanograss with high aspect ratio and needle-like tip was advantageous for improved photovoltaic conversion performance.

  2. Glioma grading using cell nuclei morphologic features in digital pathology images

    NASA Astrophysics Data System (ADS)

    Reza, Syed M. S.; Iftekharuddin, Khan M.

    2016-03-01

    This work proposes a computationally efficient cell nuclei morphologic feature analysis technique to characterize the brain gliomas in tissue slide images. In this work, our contributions are two-fold: 1) obtain an optimized cell nuclei segmentation method based on the pros and cons of the existing techniques in literature, 2) extract representative features by k-mean clustering of nuclei morphologic features to include area, perimeter, eccentricity, and major axis length. This clustering based representative feature extraction avoids shortcomings of extensive tile [1] [2] and nuclear score [3] based methods for brain glioma grading in pathology images. Multilayer perceptron (MLP) is used to classify extracted features into two tumor types: glioblastoma multiforme (GBM) and low grade glioma (LGG). Quantitative scores such as precision, recall, and accuracy are obtained using 66 clinical patients' images from The Cancer Genome Atlas (TCGA) [4] dataset. On an average ~94% accuracy from 10 fold crossvalidation confirms the efficacy of the proposed method.

  3. Biocompatible mesoporous silica nanoparticles with different morphologies for animal cell membrane penetration

    SciTech Connect

    Trewyn, B.; Nieweg, J.; Zhao, Y,; Lin, V.

    2007-11-24

    Two MCM-41 type, fluorescein-labeled mesoporous silica nanomaterials (MSNs) consisting of spherical and tube-shaped particles were synthesized and characterized. Both materials have hexagonally arranged mesopores with high surface area (>950 m{sup 2}/g) and a narrow distribution of pore diameters. The cellular uptake efficiency and kinetics of both MSNs were measured in a cancer cell line (CHO) and a noncancerous cell line (fibroblasts) by flow cytometry and fluorescence confocal microscopy. The correlation between the particle morphology and aggregation of MSNs to the effectiveness of cellular uptake was investigated. We envision that our study on the morphology dependent endocytosis of MSNs would lead to future developments of efficient transmembrane nanodevices for intracellular sensing and gene/drug delivery.

  4. Stabilization of gene expression and cell morphology after explant recycling during fin explant culture in goldfish.

    PubMed

    Chenais, Nathalie; Lareyre, Jean-Jacques; Le Bail, Pierre-Yves; Labbe, Catherine

    2015-07-01

    The development of fin primary cell cultures for in vitro cellular and physiological studies is hampered by slow cell outgrowth, low proliferation rate, poor viability, and sparse cell characterization. Here, we investigated whether the recycling of fresh explants after a first conventional culture could improve physiological stability and sustainability of the culture. The recycled explants were able to give a supplementary cell culture showing faster outgrowth, cleaner cell layers and higher net cell production. The cells exhibited a highly stabilized profile for marker gene expression including a low cytokeratin 49 (epithelial marker) and a high collagen 1a1 (mesenchymal marker) expression. Added to the cell spindle-shaped morphology, motility behavior, and actin organization, this suggests that the cells bore stable mesenchymal characteristics. This contrast with the time-evolving expression pattern observed in the control fresh explants during the first 2 weeks of culture: a sharp decrease in cytokeratin 49 expression was concomitant with a gradual increase in col1a1. We surmise that such loss of epithelial features for the benefit of mesenchymal ones was triggered by an epithelial to mesenchymal transition (EMT) process or by way of a progressive population replacement process. Overall, our findings provide a comprehensive characterization of this new primary culture model bearing mesenchymal features and whose stability over culture time makes those cells good candidates for cell reprogramming prior to nuclear transfer, in a context of fish genome preservation.

  5. Unilateral cryptorchidism induces morphological changes of testes and hyperplasia of Sertoli cells in a dog

    PubMed Central

    Moon, Joon Ho; Yoo, Dae Young; Jo, Young Kwang; Kim, Geon A; Jung, Hyo Young; Choi, Jung Hoon

    2014-01-01

    Cryptorchidism is one of the most common genital defects in dogs. This study investigated the effects of abdominal cryptorchidism on morphology, cell proliferation, and Sertoli cell condition in a dog with spontaneous unilateral cryptorchidism. Elective orchidectomy was performed on the abdominal right testis and the scrotal left testis. Significant reductions in numbers of spermatogonia, spermatocytes, and spermatids were observed in hematoxylin and eosin stained sections of the cryptorchid testis. The size of the epididymal duct was smaller than that of the control testis. Based on Ki67 immunohistochemistry, the proliferative activity of spermatogonia and spermatocytes was significantly decreased in the cryptorchid testis. However, proliferative activity was increased in the epididymal duct. Based on GATA-4 immunohistochemistry, Sertoli cells were relatively resistant to cryptorchidism, and the proliferative activity of Sertoli cells was markedly increased in the cryptorchid testis than in the control testis. These results suggest that spontaneous unilateral cryptorchidism causes morphological defects in spermatogonia and spermatocytes in the testis and changes the size of the efferent ductule of the epididymis. In addition, spontaneous unilateral cryptorchidism increases proliferative activity of Sertoli cells, which may be a predisposing factor for Sertoli cell cancer in cryptorchid testes. PMID:25628730

  6. A novel biointerface that suppresses cell morphological changes by scavenging excess reactive oxygen species.

    PubMed

    Ikeda, Yutaka; Yoshinari, Tomoki; Nagasaki, Yukio

    2015-09-01

    During cell cultivation on conventional culture dishes, various events results in strong stresses that lead to the production of bioactive species such as reactive oxygen species (ROS) and nitric oxide. These reactive species cause variable damage to cells and stimulate cellular responses. Here, we report the design of a novel biocompatible surface that decreases stress by not only morphologically modifying the dish surface by using poly(ethylene glycol) tethered chains, but also actively scavenging oxidative stress by using our novel nitroxide radical-containing polymer. A block copolymer, poly(ethylene glycol)-b-poly[(2,2,6,6-tetramethylpiperidine-N-oxyl)aminomethylstyrene] (PEG-b-PMNT) was used to coat the surface of a dish. Differentiation of undifferentiated human leukemia (HL-60) cells was found to be suppressed on the polymer-coated dish. Notably, HL-60 cell cultivation caused apoptosis under high-density conditions, while spontaneous apoptosis was suppressed in cells plated on the PEG-b-PMNT-modified surface, because a healthy mitochondrial membrane potential was maintained. In contrast, low molecular weight antioxidants did not have apparent effects on the maintenance of mitochondria. We attribute this to the lack of cellular internalization of our immobilized polymer and selective scavenging of excessive ROS generated outside of cells. These results demonstrate the utility of our novel biocompatible material for actively scavenging ROS and thus maintaining cellular morphology. PMID:25691268

  7. Primary Esophageal Extranasal NK/T Cell Lymphoma With Biphasic Morphology

    PubMed Central

    Ye, Zi-Yin; Cao, Qing-Hua; Liu, Fang; Lu, Xiao-Fang; Li, Shu-Rong; Li, Chang-Zhao; Chen, Shao-Hong

    2015-01-01

    Abstract We report a case of esophageal extranasal NK/T cell lymphoma with biphasic morphologic features revealed by a deep large piecemeal biopsy. A 40-year-old man present with pharyngalgia, dysphagia, recurrent fever, and 5-kg weight loss for 8 months. Endoscopy demonstrated progressing longitudinal ulcers and mucosal bridges along the esophagus. The first and second biopsies obtained superficial mucosa with scattered bland-looking small lymphocytes. A subsequent large piecemeal snare abscission for biopsy showed atypical lymphoid cells infiltrating into the deep lamina propria and muscularis mucosae, whereas the superficial lamina propria was highly edematous with scant small lymphocytes. Immunohistochemical studies confirmed that both underlying atypical cells and superficial small lymphocytes were neoplastic, sharing an identical immunophenotype: positive for CD2, CD3, CD43, CD8, CD56, TIA-1 and granzyme B. Epstein-Barr virus–encoded small RNAs were found in both cells. The histologic findings were diagnostic of primary esophageal extranasal NK/T cell lymphoma. However, the patient developed bone marrow depression during chemotherapy and died of massive cerebral hemorrhage after the first cycle of chemotherapy. Primary esophageal extranodal NK/T cell lymphoma nasal type is extremely rare. We show the biphasic morphology of this disease, which highlights the importance of deep biopsy for accurate diagnosis. PMID:26181557

  8. Morphologic and phenotypic changes of human neuroblastoma cells in culture induced by cytosine arabinoside

    SciTech Connect

    Ponzoni, M.; Lanciotti, M.; Melodia, A.; Casalaro, A.; Cornaglia-Ferraris, P. )

    1989-03-01

    The effects of cytosine-arabinoside (ARA-C) on the growth and phenotypic expression of a new human neuroblastoma (NB) cell line (GI-ME-N) have been extensively tested. Low doses of ARA-C allowing more than 90% cell viability induce morphological differentiation and growth inhibition. Differentiated cells were larger and flattened with elongated dendritic processes; such cells appeared within 48 hours after a dose of ARA-C as low as 0.1 {mu}g/ml. The new morphological aspect reached the maximum expression after 5-6 days of culture being independent from the addition of extra drug to the culture. A decrease in ({sup 3}H)thymidine incorporation was also observed within 24 hours and the cell growth was completely inhibited on the sixth day. Moreover, ARA-C strongly inhibited anchorage-independent growth in soft agar assay. Membrane immunofluorescence showed several dramatic changes in NB-specific antigen expression after 5 days of treatment with ARA-C. At the same time ARA-C also modulated cytoskeletal proteins and slightly increased catecholamine expression. These findings suggest that noncytotoxic doses of ARA-C do promote the differentiation of GI-ME-N neuroblastoma cells associated with reduced expression of the malignant phenotype.

  9. Monitoring of soil water storage along elevation transech on morphological diverse study-sites affected by soil erosion

    NASA Astrophysics Data System (ADS)

    Jaksik, Ondrej; Kodesova, Radka; Nikodem, Antonin; Fer, Miroslav; Klement, Ales; Kratina, Josef

    2015-04-01

    Soil water availability is one of the key factors determining plant growth. Spatial distribution of soil water content is influenced by many factors. For the field-scale, one of the most important factors is terrain and its shape. The goal of our study was to characterize soil water storage within the soil profile with respect to terrain attributes. Two morphologically diverse study sites were chosen, in order to monitor soil water storage during vegetation season. The first site Brumovice in located in the Southern Moravian Region. The original soil unit was Haplic Chernozem developed on loess, which was gradually degraded by soil erosion. In the steepest parts, due to substantial loss of soil material, soil is transformed to Regosol. As a result of consequently sedimentation of previously eroded material in toe slopes and terrain depressions colluvial soils are formed. The second site Vidim is placed in the Central Bohemia. Dominant soil unit in wider area is Haplic Luvisol on loess loam. Similar process of progressive soil transformation was identified. On each study site, two elevation transects were delimited, where each consists of 5 monitoring spots. Access tubes were installed in order to measure soil moisture in six different depths (10, 20, 30 40, 60 a 100 cm) using Profile Probe PR2. The monitoring was conducted during vegetation season: April - July 2012 in Brumovice and May - July 2013 in Vidim. The average soil water contents were calculated for following three layers: topsoil A (0-20 cm), subsoil B (20-40cm), and substrate (40-100cm). The soil water storage within the soil profile was also expressed. Sensors TMS3 were also used for continual soil water content monitoring in the depth of 0-15 cm. In addition undisturbed soil samples were taken from topsoil to measure soil hydraulic properties using the multistep outflow experiment. Data were used to assess retention ability of erosion affected soils. The soil water storage and particularly average

  10. Morphology evolution in high-performance polymer solar cells processed from nonhalogenated solvent

    DOE PAGES

    Cai, Wanzhu; Liu, Peng; Jin, Yaocheng; Xue, Qifan; Liu, Feng; Russell, Thomas P.; Huang, Fei; Yip, Hin -Lap; Cao, Yong

    2015-05-26

    A new processing protocol based on non-halogenated solvent and additive is developed to produce polymer solar cells with power conversion efficiencies better than those processed from commonly used halogenated solvent-additive pair. Morphology studies show that good performance correlates with a finely distributed nanomorphology with a well-defined polymer fibril network structure, which leads to balanced charge transport in device operation.

  11. A direct evidence of morphological degradation on a nanometer scale in polymer solar cells.

    PubMed

    Schaffer, Christoph J; Palumbiny, Claudia M; Niedermeier, Martin A; Jendrzejewski, Christian; Santoro, Gonzalo; Roth, Stephan V; Müller-Buschbaum, Peter

    2013-12-10

    In situ measurement of a polymer solar cell using micro grazing incidence small angle X-ray scattering (μGISAXS) and current-voltage tracking is demonstrated. While measuring electric characteristics under illumination, morphological changes are probed by μGISAXS. The X-ray beam (green) impinges on the photo active layer with a shallow angle and scatters on a 2d detector. Degradation is explained by the ongoing nanomorphological changes observed.

  12. [Comparison of bone marrow and blood cell morphology between refractory anemia and other anemia disease].

    PubMed

    Cheng, Hong; Jiang, Ming; DU, Wei; Zhong, Di; Hao, Jian-Ping; Li, Ling

    2012-12-01

    This study was purposed to investigate the cell morphological features of bone marrow and peripheral blood in patients with myelodysplastic syndrome, mainly with refractory anemia, and to compare them with other anemia diseases including chronic aplastic anemia, hemolytic anemia and megaloblastic anemia. The bone marrow and peripheral blood were taken from patients for preparing the smears with Wright staining. 500 karyocytes in bone marrow and 100 karyocytes in peripheral blood were detected, and the features of morbid cells of erythrocyte, granulocyte and megakaryocytic series were observed. The results showed that differences between refractory anemia, chronic aplastic anemias and hemolytic anemia as well as megaloblastic anemia were statistically significant (P < 0.05) in the granules scarce and absence in the intracytoplasm of segmented neutrocyte in peripheral blood, Pelger dyskaryosis, the numbers and detected rate of immature granulocytes, monocyte detected rate, the granules scarce in all stage of granulocytic series in bone marrow, odd number and prolification of nucleolus in erythrocytic series, little macronucleus and single circle nucleus macronucleus. It is concluded that cell morphology is the foundation of diagnosing the MDS, the abnormality morphology both in peripheral blood and bone marrow play the consequence role in the diagnosis of MDS. PMID:23257446

  13. Multifractal characterization of morphology of human red blood cells membrane skeleton.

    PubMed

    Ţălu, Ş; Stach, S; Kaczmarska, M; Fornal, M; Grodzicki, T; Pohorecki, W; Burda, K

    2016-04-01

    The purpose of this paper is to show applicability of multifractal analysis in investigations of the morphological changes of ultra-structures of red blood cells (RBCs) membrane skeleton measured using atomic force microscopy (AFM). Human RBCs obtained from healthy and hypertensive donors as well as healthy erythrocytes irradiated with neutrons (45 μGy) were studied. The membrane skeleton of the cells was imaged using AFM in a contact mode. Morphological characterization of the three-dimensional RBC surfaces was realized by a multifractal method. The nanometre scale study of human RBCs surface morphology revealed a multifractal geometry. The generalized dimensions Dq and the singularity spectrum f(α) provided quantitative values that characterize the local scale properties of their membrane skeleton organization. Surface characterization was made using areal ISO 25178-2: 2012 topography parameters in combination with AFM topography measurement. The surface structure of human RBCs is complex with hierarchical substructures resulting from the organization of the erythrocyte membrane skeleton. The analysed AFM images confirm a multifractal nature of the surface that could be useful in histology to quantify human RBC architectural changes associated with different disease states. In case of very precise measurements when the red cell surface is not wrinkled even very fine differences can be uncovered as was shown for the erythrocytes treated with a very low dose of ionizing radiation. PMID:27002485

  14. Multifractal characterization of morphology of human red blood cells membrane skeleton.

    PubMed

    Ţălu, Ş; Stach, S; Kaczmarska, M; Fornal, M; Grodzicki, T; Pohorecki, W; Burda, K

    2016-04-01

    The purpose of this paper is to show applicability of multifractal analysis in investigations of the morphological changes of ultra-structures of red blood cells (RBCs) membrane skeleton measured using atomic force microscopy (AFM). Human RBCs obtained from healthy and hypertensive donors as well as healthy erythrocytes irradiated with neutrons (45 μGy) were studied. The membrane skeleton of the cells was imaged using AFM in a contact mode. Morphological characterization of the three-dimensional RBC surfaces was realized by a multifractal method. The nanometre scale study of human RBCs surface morphology revealed a multifractal geometry. The generalized dimensions Dq and the singularity spectrum f(α) provided quantitative values that characterize the local scale properties of their membrane skeleton organization. Surface characterization was made using areal ISO 25178-2: 2012 topography parameters in combination with AFM topography measurement. The surface structure of human RBCs is complex with hierarchical substructures resulting from the organization of the erythrocyte membrane skeleton. The analysed AFM images confirm a multifractal nature of the surface that could be useful in histology to quantify human RBC architectural changes associated with different disease states. In case of very precise measurements when the red cell surface is not wrinkled even very fine differences can be uncovered as was shown for the erythrocytes treated with a very low dose of ionizing radiation.

  15. Morphology and chirality control self-assembly of sickle hemoglobin inside red blood cells

    NASA Astrophysics Data System (ADS)

    Li, Xuejin; Lei, Huan; Caswell, Bruce; Karniadakis, George

    2012-02-01

    Sickle cells exhibit abnormal morphology and membrane mechanics in the deoxygenated state due to the polymerization of the interior sickle hemoglobin (HbS). In this study, the dynamics of self-assembly behavior of HbS in solution and corresponding induced cell morphologies have been investigated by dissipative particle dynamics approach. A coarse-grained HbS model, which contains hydrophilic and hydrophobic particles, is constructed to match the structural properties and physical description (including crowding effects) of HbS. The hydrophobic interactions are shown to be necessary with chirality being the main driver for the formation of HbS fibers. In the absence of chain chirality, only the self-assembled small aggregates are observed whereas self-assembled elongated step-like bundle microstructures appear when we consider the chain chirality. Several typical cell morphologies (sickle, granular, elongated shapes), induced by the growth of HbS fibers, are revealed and their deviations from the biconcave shape are quantified by the asphericity and elliptical shape factors.

  16. A Comparison between Growth Morphology of "Eutectic" Cells/Dendrites and Single-Phase Cells/Dendrites

    NASA Technical Reports Server (NTRS)

    Tewari, S. N.; Raj, S. V.; Locci, I. E.

    2003-01-01

    Directionally solidified (DS) intermetallic and ceramic-based eutectic alloys with an in-situ composite microstructure containing finely distributed, long aspect ratio, fiber, or plate reinforcements are being seriously examined for several advanced aero-propulsion applications. In designing these alloys, additional solutes need to be added to the base eutectic composition in order to improve heir high-temperature strength, and provide for adequate toughness and resistance to environmental degradation. Solute addition, however, promotes instability at the planar liquid-solid interface resulting in the formation of two-phase eutectic "colonies." Because morphology of eutectic colonies is very similar to the single-phase cells and dendrites, the stability analysis of Mullins and Sekerka has been extended to describe their formation. Onset of their formation shows a good agreement with this approach; however, unlike the single-phase cells and dendrites, there is limited examination of their growth speed dependence of spacing, morphology, and spatial distribution. The purpose of this study is to compare the growth speed dependence of the morphology, spacing, and spatial distribution of eutectic cells and dendrites with that for the single-phase cells and dendrites.

  17. Morphological changes among hippocampal dentate granule cells exposed to early kindling-epileptogenesis.

    PubMed

    Singh, Shatrunjai P; He, Xiaoping; McNamara, James O; Danzer, Steve C

    2013-12-01

    Temporal lobe epilepsy is associated with changes in the morphology of hippocampal dentate granule cells. These changes are evident in numerous models that are associated with substantial neuron loss and spontaneous recurrent seizures. By contrast, previous studies have shown that in the kindling model, it is possible to administer a limited number of stimulations sufficient to produce a lifelong enhanced sensitivity to stimulus evoked seizures without associated spontaneous seizures and minimal neuronal loss. Here we examined whether stimulation of the amygdala sufficient to evoke five convulsive seizures (class IV or greater on Racine's scale) produce morphological changes similar to those observed in models of epilepsy associated with substantial cell loss. The morphology of GFP-expressing granule cells from Thy-1 GFP mice was examined either 1 day or 1 month after the last evoked seizure. Interestingly, significant reductions in dendritic spine density were evident 1 day after the last seizure, the magnitude of which had diminished by 1 month. Further, there was an increase in the thickness of the granule cell layer 1 day after the last evoked seizure, which was absent a month later. We also observed an increase in the area of the proximal axon, which again returned to control levels a month later. No differences in the number of basal dendrites were detected at either time point. These findings demonstrate that the early stages of kindling epileptogenesis produce transient changes in the granule cell body layer thickness, molecular layer spine density, and axon proximal area, but do not produce striking rearrangements of granule cell structure.

  18. Apoptosis in ovarian granulosa cells of cattle: morphological features and clearance by homologous phagocytosis.

    PubMed

    Carou, María Clara; Cruzans, Paula Romina; Maruri, Alejandro; Stockert, Juan Carlos; Lombardo, Daniel Marcelo

    2015-01-01

    Apoptosis is involved in many physiological processes of the ovary, such as recruitment of prenatal germ cells, follicular atresia, ovulation, and luteolysis. Based on the need for the involvement of phagocytic cells to achieve apoptosis clearance and that follicular atresia is triggered by weak apoptotic stimuli, we postulate that granulosa cells engullng apoptotic corpses (ACs) must carry out this macrophagic process. Since apoptosis was early defined in terms of morphological aspects, here we describe apoptosis induced by a GnRH analog (leuprolide acetate, LA) at histological level on bovine granulosa cells (primary culture, CPGB, and an established cell line, BGC-1). We observed two main types of apoptosis. In type A, the whole cell or most of it is compacted into a single large AC that is then engulfed by neighboring cells or simply detached. In type B, small portions of cells, either with or without nuclear material, become ACs that are also phagocytosed. Apoptosis and homologous phagocytosis were confirmed by TUNEL and immunocytochemistry for Bax and active caspase 3. Induction of apoptosis was significant in BGC-1 cells treated for 24 h with 100 nM LA. CPGB cells showed two types of response with different doses of LA. Fetal calf serum was necessary to find apoptosis induced by LA.

  19. Identification, localization and morphology of APUD cells in gastroenteropancreatic system of stomach-containing teleosts.

    PubMed

    Pan, Qian-Sheng; Fang, Zhi-Ping; Huang, Feng-Jie

    2000-12-01

    AIM:To identify the type localization and morphology of APUD endocrine cells in the gastroenteropancreatic (GEP) system of stomach-containing teleosts, and study APUD endocrine system in the stomach, intestine and pancreas of fish species.METHODS:Two kinds of immunocytochemical (ICC) techniques of the streptavidin biotin-peroxidase complex (SABC) and streptavidin-peroxidase (S-P) method were used. The identification, localization and morphology of APUD endocrine cells scattered in the mucosa of digestive tract, intermuscular nerve plexus and glandular body of northern snakehead (Channa argus), ricefield eel (Monopterus albus), yellow catfish (Pelteobagrus fulvidraco), mandarinfish (Siniperca chuatsi), largemouth bass (Micropterus salmoides),oriental sheatfish (Silurus asotus), freshwater pomfret (Colossoma brachypomum) and nile tilapia (Tilapia nilotica) were investigated with 8 kinds of antisera.RESULTS:The positive reaction of 5-hydroxytryptamine (5-HT) immunoreactive endocrine (IRE) cells was found in the digestive tract and glandular body of 8 fish species in different degree.Only a few gastrin (GAS)-IRE cells were seen in C.argus,M.albusand P.fulvidraco. Glucagon (GLU)IRE cells were not found in the digestive tract and glandular body but existed in pancreatic island of most fish species. The positive reaction of growth hormone (GH)IRE cells was found only in pancreatic island of S. Chuatsi and S. Asotus, no positive reaction in the other 6 fish species. Somatostatin (SOM), calcitonin (CAL), neurofilament (NF) and insulin (INS)-IRE cells in the stomach, intestine and pancreas of 8 kinds of fish were different in distribution and types. The distribution of all 8 APUD cells was the most in gastrointestinal epithelium mucosa and then in digestive glands. The positive reaction of SOM and 5-HT-IRE cells was found in intermuscular nerve plexus of intestine of P.fulvidraco and S.chuatsi. Only GH-IRE cells were densely scattered in the pancreatic islands of S

  20. Identification, localization and morphology of APUD cells in gastroenteropancreatic system of stomach-containing teleosts

    PubMed Central

    Pan, Qian Sheng; Fang, Zhi Ping; Huang, Feng Jie

    2000-01-01

    AIM: To identify the type localization and morphology of APUD endocrine cells in the gastroenteropancreatic (GEP) system of stomach-containing teleosts, and study APUD endocrine system in the stomach, intestine and pancreas of fish species. METHODS: Two kinds of immunocytochemical (ICC) techniques of the streptavidin biotin-peroxidase complex (SABC) and streptavidin-peroxidase (S-P) method were used. The identification, localization and morphology of APUD endocrine cells scattered in the mucosa of digestive tract, intermuscular nerve plexus and glandular body of northern snakehead (Channa argus), ricefield eel (Monopterus albus), yellow catfish (Pelteobagrus ful vidraco), mandarinfish (Siniperca chuatsi), largemouth bass (Micropterus salmoides), oriental sheatfish (Silurus asotus), freshwater pomfret (Colossoma brachypomum) and nile tilapia (Tilapia nilotica) were investigated with 8 kinds of antisera. RESULTS: The positive reaction of 5-hydroxytryptamine (5-HT) immunoreactive endocrine (IRE) cells was found in the digestive tract and glandular body of 8 fish species in different degree. Only a few gastrin (GAS)-IRE cells were seen in C. argus, M. albus and P. fulvidraco. Glucagon (GLU)-IRE cells were not found in the digestive tract and glandular body but existed in pancreatic island of most fish species. The positive reaction of growth hormone (GH)-IRE cells was found only in pancreatic island of S. Chuatsi and S. Asotus, no positive reaction in the other 6 fish species. Somatostatin (SOM), calcitonin (CAL), neurofilament (NF) and insulin (INS)-IRE cells in the stomach, intestine and pancreas of 8 kinds of fish were different in distribution and types. The distribution of all 8 APUD cells was the most in gastrointestinal epithelium mucosa and then in digestive glands. The positive reaction of SOM- and 5-HT-IRE cells was found in intermuscular nerve plexus of intestine of P. fulvidraco and S.chuatsi. Only GH-IRE cells were densely scattered in the pancreatic

  1. Electrospun fibre diameter, not alignment, affects mesenchymal stem cell differentiation into the tendon/ligament lineage.

    PubMed

    Cardwell, Robyn D; Dahlgren, Linda A; Goldstein, Aaron S

    2014-12-01

    Efforts to develop engineered tendons and ligaments have focused on the use of a biomaterial scaffold and a stem cell source. However, the ideal scaffold microenvironment to promote stem cell differentiation and development of organized extracellular matrix is unknown. Through electrospinning, fibre scaffolds can be designed with tailorable architectures to mimic the intended tissue. In this study, the effects of fibre diameter and orientation were examined by electrospinning thin mats, consisting of small (< 1 µm), medium (1-2 µm) or large (> 2 µm) diameter fibres with either random or aligned fibre orientation. C3H10T1/2 model stem cells were cultured on the six different electrospun mats, as well as smooth spin-coated films, and the morphology, growth and expression of tendon/ligament genes were evaluated. The results demonstrated that fibre diameter affects cellular behaviour more significantly than fibre alignment. Initially, cell density was greater on the small fibre diameter mats, but similar cell densities were found on all mats after an additional week in culture. After 2 weeks, gene expression of collagen 1α1 and decorin was increased on all mats compared to films. Expression of the tendon/ligament transcription factor scleraxis was suppressed on all electrospun mats relative to spin-coated films, but expression on the large-diameter fibre mats was consistently greater than on the medium-diameter fibre mats. These results suggest that larger-diameter fibres (e.g. > 2 µm) may be more suitable for in vitro development of a tendon/ligament tissue.

  2. Chalcone Synthase (CHS) Gene Suppression in Flax Leads to Changes in Wall Synthesis and Sensing Genes, Cell Wall Chemistry and Stem Morphology Parameters.

    PubMed

    Zuk, Magdalena; Działo, Magdalena; Richter, Dorota; Dymińska, Lucyna; Matuła, Jan; Kotecki, Andrzej; Hanuza, Jerzy; Szopa, Jan

    2016-01-01

    The chalcone synthase (CHS) gene controls the first step in the flavonoid biosynthesis. In flax, CHS down-regulation resulted in tannin accumulation and reduction in lignin synthesis, but plant growth was not affected. This suggests that lignin content and thus cell wall characteristics might be modulated through CHS activity. This study investigated the possibility that CHS affects cell wall sensing as well as polymer content and arrangement. CHS-suppressed and thus lignin-reduced plants showed significant changes in expression of genes involved in both synthesis of components and cell wall sensing. This was accompanied by increased levels of cellulose and hemicellulose. CHS-reduced flax also showed significant changes in morphology and arrangement of the cell wall. The stem tissue layers were enlarged averagely twofold compared to the control, and the number of fiber cells more than doubled. The stem morphology changes were accompanied by reduction of the crystallinity index of the cell wall. CHS silencing induces a signal transduction cascade that leads to modification of plant metabolism in a wide range and thus cell wall structure. PMID:27446124

  3. Chalcone Synthase (CHS) Gene Suppression in Flax Leads to Changes in Wall Synthesis and Sensing Genes, Cell Wall Chemistry and Stem Morphology Parameters

    PubMed Central

    Zuk, Magdalena; Działo, Magdalena; Richter, Dorota; Dymińska, Lucyna; Matuła, Jan; Kotecki, Andrzej; Hanuza, Jerzy; Szopa, Jan

    2016-01-01

    The chalcone synthase (CHS) gene controls the first step in the flavonoid biosynthesis. In flax, CHS down-regulation resulted in tannin accumulation and reduction in lignin synthesis, but plant growth was not affected. This suggests that lignin content and thus cell wall characteristics might be modulated through CHS activity. This study investigated the possibility that CHS affects cell wall sensing as well as polymer content and arrangement. CHS-suppressed and thus lignin-reduced plants showed significant changes in expression of genes involved in both synthesis of components and cell wall sensing. This was accompanied by increased levels of cellulose and hemicellulose. CHS-reduced flax also showed significant changes in morphology and arrangement of the cell wall. The stem tissue layers were enlarged averagely twofold compared to the control, and the number of fiber cells more than doubled. The stem morphology changes were accompanied by reduction of the crystallinity index of the cell wall. CHS silencing induces a signal transduction cascade that leads to modification of plant metabolism in a wide range and thus cell wall structure. PMID:27446124

  4. Chalcone Synthase (CHS) Gene Suppression in Flax Leads to Changes in Wall Synthesis and Sensing Genes, Cell Wall Chemistry and Stem Morphology Parameters.

    PubMed

    Zuk, Magdalena; Działo, Magdalena; Richter, Dorota; Dymińska, Lucyna; Matuła, Jan; Kotecki, Andrzej; Hanuza, Jerzy; Szopa, Jan

    2016-01-01

    The chalcone synthase (CHS) gene controls the first step in the flavonoid biosynthesis. In flax, CHS down-regulation resulted in tannin accumulation and reduction in lignin synthesis, but plant growth was not affected. This suggests that lignin content and thus cell wall characteristics might be modulated through CHS activity. This study investigated the possibility that CHS affects cell wall sensing as well as polymer content and arrangement. CHS-suppressed and thus lignin-reduced plants showed significant changes in expression of genes involved in both synthesis of components and cell wall sensing. This was accompanied by increased levels of cellulose and hemicellulose. CHS-reduced flax also showed significant changes in morphology and arrangement of the cell wall. The stem tissue layers were enlarged averagely twofold compared to the control, and the number of fiber cells more than doubled. The stem morphology changes were accompanied by reduction of the crystallinity index of the cell wall. CHS silencing induces a signal transduction cascade that leads to modification of plant metabolism in a wide range and thus cell wall structure.

  5. An epilepsy-related ARX polyalanine expansion modifies glutamatergic neurons excitability and morphology without affecting GABAergic neurons development.

    PubMed

    Beguin, Shirley; Crépel, Valérie; Aniksztejn, Laurent; Becq, Hélène; Pelosi, Barbara; Pallesi-Pocachard, Emilie; Bouamrane, Lamine; Pasqualetti, Massimo; Kitamura, Kunio; Cardoso, Carlos; Represa, Alfonso

    2013-06-01

    Epileptic encephalopathies comprise a heterogeneous group of severe infantile disorders for which the pathophysiological basis of epilepsy is inaccurately clarified by genotype-phenotype analysis. Because a deficit of GABA neurons has been found in some of these syndromes, notably in patients with X-linked lissencephaly with abnormal genitalia, epilepsy was suggested to result from an imbalance in GABAergic inhibition, and the notion of "interneuronopathy" was proposed. Here, we studied the impact of a polyalanine expansion of aristaless-related homeobox (ARX) gene, a mutation notably found in West and Ohtahara syndromes. Analysis of Arx((GCG)7/Y) knock-in mice revealed that GABA neuron development is not affected. Moreover, pyramidal cell migration and cortical layering are unaltered in these mice. Interestingly, electrophysiological recordings show that hippocampal pyramidal neurons displayed a frequency of inhibitory postsynaptic currents similar to wild-type (WT) mice. However, these neurons show a dramatic increase in the frequency of excitatory inputs associated with a remodeling of their axonal arborization, suggesting that epilepsy in Arx((GCG)7/Y)mice would result from a glutamate network remodeling. We therefore propose that secondary alterations are instrumental for the development of disease-specific phenotypes and should be considered to explain the phenotypic diversity associated with epileptogenic mutations. PMID:22628459

  6. Identification and quantitation of morphological cell types in electrophoretically separated human embryonic kidney cell cultures

    NASA Technical Reports Server (NTRS)

    Williams, K. B.; Kunze, M. E.; Todd, P. W.

    1985-01-01

    Four major cell types were identified by phase microscopy in early passage human embryonic kidney cell cultures. They are small and large epithelioid, domed, and fenestrated cells. Fibroblasts are also present in some explants. The percent of each cell type changes with passage number as any given culture grows. As a general rule, the fraction of small epithelioid cells increases, while the fraction of fenestrated cells, always small, decreases further. When fibroblasts are present, they always increase in percentage of the total cell population. Electrophoretic separation of early passage cells showed that the domed cells have the highest electrophoretic mobility, fibroblasts have an intermediate high mobility, small epithelioid cells have a low mobility, broadly distributed, and fenestrated cells have the lowest mobility. All cell types were broadly distributed among electrophoretic subfractions, which were never pure but only enriched with respect to a given cell type.

  7. From nano to micro: topographical scale and its impact on cell adhesion, morphology and contact guidance

    NASA Astrophysics Data System (ADS)

    Nguyen, Anh Tuan; Sathe, Sharvari R.; Yim, Evelyn K. F.

    2016-05-01

    Topography, among other physical factors such as substrate stiffness and extracellular forces, is known to have a great influence on cell behaviours. Optimization of topographical features, in particular topographical dimensions ranging from nanoscale to microscale, is the key strategy to obtain the best cellular performance for various applications in tissue engineering and regenerative medicine. In this review, we provide a comprehensive survey on the significance of sizes of topography and their impacts on cell adhesion, morphology and alignment, and neurite guidance. Also recent works mimicking the hierarchical structure of natural extracellular matrix by combining both nanoscale and microscale topographies are highlighted.

  8. Assessing epithelial cell nuclear morphology by using azimuthal light scattering spectroscopy.

    PubMed

    Yu, Chung-Chieh; Lau, Condon; Tunnell, James W; Hunter, Martin; Kalashnikov, Maxim; Fang-Yen, Christopher; Fulghum, Stephen F; Badizadegan, Kamran; Dasari, Ramachandra R; Feld, Michael S

    2006-11-01

    We describe azimuthal light scattering spectroscopy (phi/LSS), a novel technique for assessing epithelial-cell nuclear morphology. The difference between the spectra measured at azimuthal angles phi = 0 degrees and phi = 90 degrees preferentially isolates the single backscattering contribution due to large (approximately 10 microm) structures such as epithelial cell nuclei by discriminating against scattering from smaller organelles and diffusive background. We demonstrate the feasibility of using phi/LSS for cancer detection by showing that spectra from cancerous colon tissue exhibit significantly greater azimuthal asymmetry than spectra from normal colonic tissues. PMID:17041654

  9. A photocurable hydrogel/elastomer composite scaffold with bi-continuous morphology for cell encapsulation.

    PubMed

    Hayami, James W S; Waldman, Stephen D; Amsden, Brian G

    2011-12-01

    A photocurable two-phase scaffold with a bi-continuous morphology was designed and characterized for the repair of load bearing soft tissues. An N-methacrylate glycol chitosan (MGC) hydrogel phase was used to distribute the cells and enable cell growth once crosslinked. The second phase, an elastomerprepared from a star-poly(ε-caprolactone-co-D,L-lactide) triacrylate, was used to enhance the mechanical properties. Chondrocytes were photocrosslinked within the bi-continuous scaffolds and proliferated, increased metabolic activity and accumulated extracellular matrix over a 14 d culture period. Also during this time no significant material degradation was observed. PMID:22012746

  10. Primary culture of endothelial cells from atherosclerotic human aorta. Part 1. Identification, morphological and ultrastructural characteristics of two endothelial cell subpopulations.

    PubMed

    Antonov, A S; Nikolaeva, M A; Klueva, T S; Romanov YuA; Babaev, V R; Bystrevskaya, V B; Perov, N A; Repin, V S; Smirnov, V N

    1986-01-01

    Endothelial cells (EC) were harvested by 0.1% collagenase treatment for adult human thoracic aortas obtained 1-3 h after sudden death. At least 35-70% of EC were removed from the intimal surface of aorta, 90-95% of them being viable. Plating efficiency was 70-80%. Monolayer formation was achieved at a seeding density of 5-8 X 10(2) cells/mm2. The cells were identified as endothelium by the presence of Factor VIII antigen, Weigel-Palade bodies and typically endothelial morphology at confluence. Unlike endothelial cultures derived from human umbilical veins and infant aortas, primary cultures obtained from human adult aortas contain multinuclear EC with Factor VIII antigen and Weibel-Palade bodies. The number of multinuclear EC in cultures isolated from aortas affected by atherosclerosis was 2-fold higher (P less than 0.05) than in cultures obtained from grossly normal aortas taken from donors of the same age. EC with numerous lipid inclusions revealed by oil-red-O staining were present in all the EC primary cultures derived from aortas affected by atherosclerosis. No oil-red-O-positive cells were detected among the EC cultured from infant aorta, aorta of young donors, and umbilical vein. An electron microscopic examination of EC from atherosclerotic aorta in culture and in situ failed to reveal any ultrastructural peculiarities distinguishing multinuclear EC from the mononuclear EC. PMID:3004520

  11. FoxP2 protein levels regulate cell morphology changes and migration patterns in the vertebrate developing telencephalon.

    PubMed

    Garcia-Calero, Elena; Botella-Lopez, Arancha; Bahamonde, Olga; Perez-Balaguer, Ariadna; Martinez, Salvador

    2016-07-01

    In the mammalian telencephalon, part of the progenitor cells transition from multipolar to bipolar morphology as they invade the mantle zone. This associates with changing patterns of radial migration. However, the molecules implicated in these morphology transitions are not well known. In the present work, we analyzed the function of FoxP2 protein in this process during telencephalic development in vertebrates. We analyzed the expression of FoxP2 protein and its relation with cell morphology and migratory patterns in mouse and chicken developing striatum. We observed FoxP2 protein expressed in a gradient from the subventricular zone to the mantle layer in mice embryos. In the FoxP2 low domain cells showed multipolar migration. In the striatal mantle layer where FoxP2 protein expression is higher, cells showed locomoting migration and bipolar morphology. In contrast, FoxP2 showed a high and homogenous expression pattern in chicken striatum, thus bipolar morphology predominated. Elevation of FoxP2 in the striatal subventricular zone by in utero electroporation promoted bipolar morphology and impaired multipolar radial migration. In mouse cerebral cortex we obtained similar results. FoxP2 promotes transition from multipolar to bipolar morphology by means of gradiental expression in mouse striatum and cortex. Together these results indicate a role of FoxP2 differential expression in cell morphology control of the vertebrate telencephalon.

  12. Morphological Variability and Distinct Protein Profiles of Cultured and Endosymbiotic Symbiodinium cells Isolated from Exaiptasia pulchella

    PubMed Central

    Pasaribu, Buntora; Weng, Li-Chi; Lin, I-Ping; Camargo, Eddie; Tzen, Jason T. C.; Tsai, Ching-Hsiu; Ho, Shin-Lon; Lin, Mong-Rong; Wang, Li-Hsueh; Chen, Chii-Shiarng; Jiang, Pei-Luen

    2015-01-01

    Symbiodinium is a dinoflagellate that plays an important role in the physiology of the symbiotic relationships of Cnidarians such as corals and sea anemones. However, it is very difficult to cultivate free-living dinoflagellates after being isolated from the host, as they are very sensitive to environmental changes. How these symbiont cells are supported by the host tissue is still unclear. This study investigated the characteristics of Symbiodinium cells, particularly with respect to the morphological variability and distinct protein profiles of both cultured and endosymbiotic Symbiodinium which were freshly isolated from Exaiptasia pulchella. The response of the cellular morphology of freshly isolated Symbiodinium cells kept under a 12 h L:12 h D cycle to different temperatures was measured. Cellular proliferation was investigated by measuring the growth pattern of Symbiodinium cells, the results of which indicated that the growth was significantly reduced in response to the extreme temperatures. Proteomic analysis of freshly isolated Symbiodinium cells revealed twelve novel proteins that putatively included transcription translation factors, photosystem proteins, and proteins associated with energy and lipid metabolism, as well as defense response. The results of this study will bring more understandings to the mechanisms governing the endosymbiotic relationship between the cnidarians and dinoflagellates. PMID:26481560

  13. Morphological Variability and Distinct Protein Profiles of Cultured and Endosymbiotic Symbiodinium cells Isolated from Exaiptasia pulchella

    NASA Astrophysics Data System (ADS)

    Pasaribu, Buntora; Weng, Li-Chi; Lin, I.-Ping; Camargo, Eddie; Tzen, Jason T. C.; Tsai, Ching-Hsiu; Ho, Shin-Lon; Lin, Mong-Rong; Wang, Li-Hsueh; Chen, Chii-Shiarng; Jiang, Pei-Luen

    2015-10-01

    Symbiodinium is a dinoflagellate that plays an important role in the physiology of the symbiotic relationships of Cnidarians such as corals and sea anemones. However, it is very difficult to cultivate free-living dinoflagellates after being isolated from the host, as they are very sensitive to environmental changes. How these symbiont cells are supported by the host tissue is still unclear. This study investigated the characteristics of Symbiodinium cells, particularly with respect to the morphological variability and distinct protein profiles of both cultured and endosymbiotic Symbiodinium which were freshly isolated from Exaiptasia pulchella. The response of the cellular morphology of freshly isolated Symbiodinium cells kept under a 12 h L:12 h D cycle to different temperatures was measured. Cellular proliferation was investigated by measuring the growth pattern of Symbiodinium cells, the results of which indicated that the growth was significantly reduced in response to the extreme temperatures. Proteomic analysis of freshly isolated Symbiodinium cells revealed twelve novel proteins that putatively included transcription translation factors, photosystem proteins, and proteins associated with energy and lipid metabolism, as well as defense response. The results of this study will bring more understandings to the mechanisms governing the endosymbiotic relationship between the cnidarians and dinoflagellates.

  14. Morphological Variability and Distinct Protein Profiles of Cultured and Endosymbiotic Symbiodinium cells Isolated from Exaiptasia pulchella.

    PubMed

    Pasaribu, Buntora; Weng, Li-Chi; Lin, I-Ping; Camargo, Eddie; Tzen, Jason T C; Tsai, Ching-Hsiu; Ho, Shin-Lon; Lin, Mong-Rong; Wang, Li-Hsueh; Chen, Chii-Shiarng; Jiang, Pei-Luen

    2015-10-20

    Symbiodinium is a dinoflagellate that plays an important role in the physiology of the symbiotic relationships of Cnidarians such as corals and sea anemones. However, it is very difficult to cultivate free-living dinoflagellates after being isolated from the host, as they are very sensitive to environmental changes. How these symbiont cells are supported by the host tissue is still unclear. This study investigated the characteristics of Symbiodinium cells, particularly with respect to the morphological variability and distinct protein profiles of both cultured and endosymbiotic Symbiodinium which were freshly isolated from Exaiptasia pulchella. The response of the cellular morphology of freshly isolated Symbiodinium cells kept under a 12 h L:12 h D cycle to different temperatures was measured. Cellular proliferation was investigated by measuring the growth pattern of Symbiodinium cells, the results of which indicated that the growth was significantly reduced in response to the extreme temperatures. Proteomic analysis of freshly isolated Symbiodinium cells revealed twelve novel proteins that putatively included transcription translation factors, photosystem proteins, and proteins associated with energy and lipid metabolism, as well as defense response. The results of this study will bring more understandings to the mechanisms governing the endosymbiotic relationship between the cnidarians and dinoflagellates.

  15. Phenomenology based multiscale models as tools to understand cell membrane and organelle morphologies

    PubMed Central

    Ramakrishnan, N.; Radhakrishnan, Ravi

    2016-01-01

    An intriguing question in cell biology is “how do cells regulate their shape?” It is commonly believed that the observed cellular morphologies are a result of the complex interaction among the lipid molecules (constituting the cell membrane), and with a number of other macromolecules, such as proteins. It is also believed that the common biophysical processes essential for the functioning of a cell also play an important role in cellular morphogenesis. At the cellular scale—where typical dimensions are in the order of micrometers—the effects arising from the molecular scale can either be modeled as equilibrium or non-equilibrium processes. In this chapter, we discuss the dynamically triangulated Monte Carlo technique to model and simulate membrane morphologies at the cellular scale, which in turn can be used to investigate several questions related to shape regulation in cells. In particular, we focus on two specific problems within the framework of isotropic and anisotropic elasticity theories: namely, (i) the origin of complex, physiologically relevant, membrane shapes due to the interaction of the membrane with curvature remodeling proteins, and (ii) the genesis of steady state cellular shapes due to the action of non-equilibrium forces that are generated by the fission and fusion of transport vesicles and by the binding and unbinding of proteins from the parent membrane. PMID:27087801

  16. Morphological Variability and Distinct Protein Profiles of Cultured and Endosymbiotic Symbiodinium cells Isolated from Exaiptasia pulchella.

    PubMed

    Pasaribu, Buntora; Weng, Li-Chi; Lin, I-Ping; Camargo, Eddie; Tzen, Jason T C; Tsai, Ching-Hsiu; Ho, Shin-Lon; Lin, Mong-Rong; Wang, Li-Hsueh; Chen, Chii-Shiarng; Jiang, Pei-Luen

    2015-01-01

    Symbiodinium is a dinoflagellate that plays an important role in the physiology of the symbiotic relationships of Cnidarians such as corals and sea anemones. However, it is very difficult to cultivate free-living dinoflagellates after being isolated from the host, as they are very sensitive to environmental changes. How these symbiont cells are supported by the host tissue is still unclear. This study investigated the characteristics of Symbiodinium cells, particularly with respect to the morphological variability and distinct protein profiles of both cultured and endosymbiotic Symbiodinium which were freshly isolated from Exaiptasia pulchella. The response of the cellular morphology of freshly isolated Symbiodinium cells kept under a 12 h L:12 h D cycle to different temperatures was measured. Cellular proliferation was investigated by measuring the growth pattern of Symbiodinium cells, the results of which indicated that the growth was significantly reduced in response to the extreme temperatures. Proteomic analysis of freshly isolated Symbiodinium cells revealed twelve novel proteins that putatively included transcription translation factors, photosystem proteins, and proteins associated with energy and lipid metabolism, as well as defense response. The results of this study will bring more understandings to the mechanisms governing the endosymbiotic relationship between the cnidarians and dinoflagellates. PMID:26481560

  17. Cystic Renal Oncocytoma and Tubulocystic Renal Cell Carcinoma: Morphologic and Immunohistochemical Comparative Study.

    PubMed

    Skenderi, Faruk; Ulamec, Monika; Vranic, Semir; Bilalovic, Nurija; Peckova, Kvetoslava; Rotterova, Pavla; Kokoskova, Bohuslava; Trpkov, Kiril; Vesela, Pavla; Hora, Milan; Kalusova, Kristyna; Sperga, Maris; Perez Montiel, Delia; Alvarado Cabrero, Isabel; Bulimbasic, Stela; Branzovsky, Jindrich; Michal, Michal; Hes, Ondrej

    2016-02-01

    Renal oncocytoma (RO) may present with a tubulocystic growth in 3% to 7% of cases, and in such cases its morphology may significantly overlap with tubulocystic renal cell carcinoma (TCRCC). We compared the morphologic and immunohistochemical characteristics of these tumors, aiming to clarify the differential diagnostic criteria, which facilitate the discrimination of RO from TCRCC. Twenty-four cystic ROs and 15 TCRCCs were selected and analyzed for: architectural growth patterns, stromal features, cytomorphology, ISUP nucleolar grade, necrosis, and mitotic activity. Immunohistochemical panel included various cytokeratins (AE1-AE3, OSCAR, CAM5.2, CK7), vimentin, CD10, CD117, AMACR, CA-IX, antimitochondrial antigen (MIA), EMA, and Ki-67. The presence of at least focal solid growth and islands of tumor cells interspersed with loose stroma, lower ISUP nucleolar grade, absence of necrosis, and absence of mitotic figures were strongly suggestive of a cystic RO. In contrast, the absence of solid and island growth patterns and presence of more compact, fibrous stroma, accompanied by higher ISUP nucleolar grade, focal necrosis, and mitotic figures were all associated with TCRCC. TCRCC marked more frequently for vimentin, CD10, AMACR, and CK7 and had a higher proliferative index by Ki-67 (>15%). CD117 was negative in 14/15 cases. One case was weakly CD117 reactive with cytoplasmic positivity. All cystic RO cases were strongly positive for CD117. The remaining markers (AE1-AE3, CAM5.2, OSCAR, CA-IX, MIA, EMA) were of limited utility. Presence of tumor cell islands and solid growth areas and the type of stroma may be major morphologic criteria in differentiating cystic RO from TCRCC. In difficult cases, or when a limited tissue precludes full morphologic assessment, immunohistochemical pattern of vimentin, CD10, CD117, AMACR, CK7, and Ki-67 could help in establishing the correct diagnosis. PMID:26180933

  18. Comparative morphology of dendritic arbors in populations of Purkinje cells in mouse sulcus and apex.

    PubMed

    Nedelescu, Hermina; Abdelhack, Mohamed

    2013-01-01

    Foliation divides the mammalian cerebellum into structurally distinct subdivisions, including the concave sulcus and the convex apex. Purkinje cell (PC) dendritic morphology varies between subdivisions and changes significantly ontogenetically. Since dendritic morphology both enables and limits sensory-motor circuit function, it is important to understand how neuronal architectures differ between brain regions. This study employed quantitative confocal microcopy to reconstruct dendritic arbors of cerebellar PCs expressing green fluorescent protein and compared arbor morphology between PCs of sulcus and apex in young and old mice. Arbors were digitized from high z-resolution (0.25 µm) image stacks using an adaptation of Neurolucida's (MBF Bioscience) continuous contour tracing tool, designed for drawing neuronal somata. Reconstructed morphologies reveal that dendritic arbors of sulcus and apex exhibit profound differences. In sulcus, 72% of the young PC population possesses two primary dendrites, whereas in apex, only 28% do. Spatial constraints in the young sulcus cause significantly more dendritic arbor overlap than in young apex, a distinction that disappears in adulthood. However, adult sulcus PC arbors develop a greater number of branch crossings. These results suggest developmental neuronal plasticity that enables cerebellar PCs to attain correct functional adult architecture under different spatial constraints.

  19. Zinc air refuelable battery: alternative zinc fuel morphologies and cell behavior

    SciTech Connect

    Cooper, J.F.; Krueger, R.

    1997-01-01

    Multicell zinc/air batteries have been tested previously in the laboratory and as part of the propulsion system of an electric bus; cut zinc wire was used as the anode material. This battery is refueled by a hydraulic transport of 0.5-1 mm zinc particles into hoppers above each cell. We report an investigation concerning alternative zinc fuel morphologies, and energy losses associated with refueling and with overnight or prolonged standby. Three types of fuel pellets were fabricated, tested and compared with results for cut wire: spheres produced in a fluidized bed electrolysis cell; elongated particles produced by gas-atomization; and pellets produced by chopping 1 mm porous plates made of compacted zinc fines. Relative sizes of the particles and cell gap dimensions are critical. All three types transported within the cell 1553 and showed acceptable discharge characteristics, but a fluidized bed approach appears especially attractive for owner/user recovery operations.

  20. [Morphologic characteristics of plaques produced by the rubella virus in cultures of Vero cells].

    PubMed

    Nates, S V; Márquez, A; Zapata, M

    1986-01-01

    Under defined plaquing condition, Gilschrist strain of rubella virus showed different plaque morphology and plaque size when tested in Vero cell cultures. These differences were obtained by changing the fetal bovine serum (FBS) concentration. With 2 per cent of FBS rubella virus formed clear plaques that included a number of cells which retained the stain, while with 4 per cent of FBS it formed ring-shaped plaques. These characteristics were retained even after the Gilschrist strain was passaged several times in Vero cell cultures. The cytopathic effect in Vero cell cultures proved to be useful for the titration of rubella virus, giving infective titres in the same logarithmic order than the plaque assay.

  1. Morphologic and cytochemical characteristics of green turtle (Chelonia mydas) blood cells

    USGS Publications Warehouse

    Work, T.M.; Raskin, R.E.; Balazs, G.H.; Whittaker, S.D.

    1998-01-01

    Objective - To identify and characterize blood cells from free-ranging Hawaiian green turtles, Chelonia mydas. Sample Population - 26 green turtles from Puako on the island of Hawaii and Kaneohe Bay on the island of Oahu. Procedure - Blood was examined, using light and electron microscopy and cytochemical stains that included benzidine peroxidase, chloroacetate esterase, alpha naphthyl butyrate esterase, acid phosphatase, Sudan black B, periodic acid-Schiff, and toluidine blue. Results - 6 types of WBC were identified: lymphocytes, monocytes, thrombocytes, heterophils, basophils, and eosinophils (small and large). Morphologic characteristics of mononuclear cells and most granulocytes were similar to those of cells from other reptiles except that green turtles have both large and small eosinophils. Conclusions - Our classification of green turtle blood cells clarifies imporoper nomenclature reported previously and provides a reference for future hematologic studies in this species.

  2. [The dynamic morphology of the endothelial cells of aortocoronary vessels cultured in vitro].

    PubMed

    Martín del Campo, D; Gómez, M L; Chévez, A

    1992-01-01

    The morphologic and functional meaning of the endothelial cell, as the first cellular element between blood and tissues, has been investigated since the earliest histological knowledge of the vascular tree. Although there have been multiple attempts for its study, through different biological branches, very little has been comprehended in its dynamic biology, due to technical difficulties and to the apparent simplicity of this extraordinary cellular type. In this research we pretend to establish a morphodynamic pattern of the endothelial cells from the aortocoronary segment, using the particular advantages of tissue culture. Our results, have been obtained through careful observations with light microscopy under different optical systems and the help provided by spaced microcinematography. Our results showed the reality of the images in the different cellular phenomena, particularly, endocytotic pinocytosis, the dynamic of the cellular membrane, and the cell-cell linkages established in vitro.

  3. Principles of connectivity among morphologically defined cell types in adult neocortex.

    PubMed

    Jiang, Xiaolong; Shen, Shan; Cadwell, Cathryn R; Berens, Philipp; Sinz, Fabian; Ecker, Alexander S; Patel, Saumil; Tolias, Andreas S

    2015-11-27

    Since the work of Ramón y Cajal in the late 19th and early 20th centuries, neuroscientists have speculated that a complete understanding of neuronal cell types and their connections is key to explaining complex brain functions. However, a complete census of the constituent cell types and their wiring diagram in mature neocortex remains elusive. By combining octuple whole-cell recordings with an optimized avidin-biotin-peroxidase staining technique, we carried out a morphological and electrophysiological census of neuronal types in layers 1, 2/3, and 5 of mature neocortex and mapped the connectivity between more than 11,000 pairs of identified neurons. We categorized 15 types of interneurons, and each exhibited a characteristic pattern of connectivity with other interneuron types and pyramidal cells. The essential connectivity structure of the neocortical microcircuit could be captured by only a few connectivity motifs.

  4. Target morphology and cell memory: a model of regenerative pattern formation.

    PubMed

    Bessonov, Nikolai; Levin, Michael; Morozova, Nadya; Reinberg, Natalia; Tosenberger, Alen; Volpert, Vitaly

    2015-12-01

    Despite the growing body of work on molecular components required for regenerative repair, we still lack a deep understanding of the ability of some animal species to regenerate their appropriate complex anatomical structure following damage. A key question is how regenerating systems know when to stop growth and remodeling - what mechanisms implement recognition of correct morphology that signals a stop condition? In this work, we review two conceptual models of pattern regeneration that implement a kind of pattern memory. In the first one, all cells communicate with each other and keep the value of the total signal received from the other cells. If a part of the pattern is amputated, the signal distribution changes. The difference fromthe original signal distribution stimulates cell proliferation and leads to pattern regeneration, in effect implementing an error minimization process that uses signaling memory to achieve pattern correction. In the second model, we consider a more complex pattern organization with different cell types. Each tissue contains a central (coordinator) cell that controls the tissue and communicates with the other central cells. Each of them keeps memory about the signals received from other central cells. The values of these signals depend on the mutual cell location, and the memory allows regeneration of the structure when it is modified. The purpose of these models is to suggest possible mechanisms of pattern regeneration operating on the basis of cell memory which are compatible with diverse molecular implementation mechanisms within specific organisms. PMID:26889161

  5. Target morphology and cell memory: a model of regenerative pattern formation

    PubMed Central

    Bessonov, Nikolai; Levin, Michael; Morozova, Nadya; Reinberg, Natalia; Tosenberger, Alen; Volpert, Vitaly

    2015-01-01

    Despite the growing body of work on molecular components required for regenerative repair, we still lack a deep understanding of the ability of some animal species to regenerate their appropriate complex anatomical structure following damage. A key question is how regenerating systems know when to stop growth and remodeling – what mechanisms implement recognition of correct morphology that signals a stop condition? In this work, we review two conceptual models of pattern regeneration that implement a kind of pattern memory. In the first one, all cells communicate with each other and keep the value of the total signal received from the other cells. If a part of the pattern is amputated, the signal distribution changes. The difference fromthe original signal distribution stimulates cell proliferation and leads to pattern regeneration, in effect implementing an error minimization process that uses signaling memory to achieve pattern correction. In the second model, we consider a more complex pattern organization with different cell types. Each tissue contains a central (coordinator) cell that controls the tissue and communicates with the other central cells. Each of them keeps memory about the signals received from other central cells. The values of these signals depend on the mutual cell location, and the memory allows regeneration of the structure when it is modified. The purpose of these models is to suggest possible mechanisms of pattern regeneration operating on the basis of cell memory which are compatible with diverse molecular implementation mechanisms within specific organisms. PMID:26889161

  6. mGluR5 Positive and Negative Allosteric Modulators Differentially Affect Dendritic Spine Density and Morphology in the Prefrontal Cortex.

    PubMed

    LaCrosse, Amber L; Taylor, Sara B; Nemirovsky, Natali E; Gass, Justin T; Olive, Michael F

    2015-01-01

    Positive and negative allosteric modulators (PAMs and NAMs, respectively) of type 5 metabotropic glutamate receptors (mGluR5) are currently being investigated as novel treatments for neuropsychiatric diseases including drug addiction, schizophrenia, and Fragile X syndrome. However, only a handful of studies have examined the effects of mGluR5 PAMs or NAMs on the structural plasticity of dendritic spines in otherwise naïve animals, particularly in brain regions mediating executive function. In the present study, we assessed dendritic spine density and morphology in pyramidal cells of the medial prefrontal cortex (mPFC) after repeated administration of either the prototypical mGluR5 PAM 3-cyano-N-(1,3-diphenyl-1H-pyrazol-5- yl)benzamide (CDPPB, 20 mg/kg), the clinically utilized mGluR5 NAM 1-(3-chlorophenyl)-3-(3-methyl-5-oxo-4Himidazol- 2-yl)urea (fenobam, 20 mg/kg), or vehicle in male Sprague-Dawley rats. Following once daily treatment for 10 consecutive days, coronal brain sections containing the mPFC underwent diolistic labeling and 3D image analysis of dendritic spines. Compared to vehicle treated animals, rats administered fenobam exhibited significant increases in dendritic spine density and the overall frequency of spines with small (<0.2 μm) head diameters, decreases in frequency of spines with medium (0.2-0.4 μm) head diameters, and had no changes in frequency of spines with large head diameters (>0.4 μm). Administration of CDPPB had no discernable effects on dendritic spine density or morphology, and neither CDPPB nor fenobam had any effect on spine length or volume. We conclude that mGluR5 PAMs and NAMs differentially affect mPFC dendritic spine structural plasticity in otherwise naïve animals, and additional studies assessing their effects in combination with cognitive or behavioral tasks are needed. PMID:25921744

  7. mGluR5 Positive and Negative Allosteric Modulators Differentially Affect Dendritic Spine Density and Morphology in the Prefrontal Cortex

    PubMed Central

    LaCrosse, Amber L.; Taylor, Sara B.; Nemirovsky, Natali E.; Gass, Justin T.; Olive, M. Foster

    2015-01-01

    Positive and negative allosteric modulators (PAMs and NAMs, respectively) of type 5 metabotropic glutamate receptors (mGluR5) are currently being investigated as novel treatments for neuropsychiatric diseases including drug addiction, schizophrenia, and Fragile X syndrome. However, only a handful of studies have examined the effects of mGluR5 PAMs or NAMs on the structural plasticity of dendritic spines in otherwise naïve animals, particularly in brain regions mediating executive function. In the present study, we assessed dendritic spine density and morphology in pyramidal cells of the medial prefrontal cortex (mPFC) after repeated administration of either the prototypical mGluR5 PAM 3-cyano-N-(1,3-diphenyl-1H-pyrazol-5-yl)benzamide (CDPPB, 20 mg/kg), the clinically utilized mGluR5 NAM 1-(3-chlorophenyl)-3-(3-methyl-5-oxo-4H-imidazol-2-yl)urea (fenobam, 20 mg/kg), or vehicle in male Sprague-Dawley rats. Following once daily treatment for 10 consecutive days, coronal brain sections containing the mPFC underwent diolistic labeling and 3D image analysis of dendritic spines. Compared to vehicle treated animals, rats administered fenobam exhibited significant increases in dendritic spine density and the overall frequency of spines with small (<0.2 μm) head diameters, decreases in frequency of spines with medium (0.2–0.4 μm) head diameters, and had no changes in frequency of spines with large head diameters (>0.4 μm). Administration of CDPPB had no discernable effects on dendritic spine density or morphology, and neither CDPPB nor fenobam had any effect on spine length or volume. We conclude that mGluR5 PAMs and NAMs differentially affect mPFC dendritic spine structural plasticity in otherwise naïve animals, and additional studies assessing their effects in combination with cognitive or behavioral tasks are needed. PMID:25921744

  8. Morphological Characteristics of Electrophysiologically Characterized Layer Vb Pyramidal Cells in Rat Barrel Cortex

    PubMed Central

    Loucif, Alexandre J. C.; Schubert, Dirk; Möck, Martin

    2016-01-01

    Layer Vb pyramidal cells are the major output neurons of the neocortex and transmit the outcome of cortical columnar signal processing to distant target areas. At the same time they contribute to local tactile information processing by emitting recurrent axonal collaterals into the columnar microcircuitry. It is, however, not known how exactly the two types of pyramidal cells, called slender-tufted and thick-tufted, contribute to the local circuitry. Here, we investigated in the rat barrel cortex the detailed quantitative morphology of biocytin-filled layer Vb pyramidal cells in vitro, which were characterized for their intrinsic electrophysiology with special emphasis on their action potential firing pattern. Since we stained the same slices for cytochrome oxidase, we could also perform layer- and column-related analyses. Our results suggest that in layer Vb the unambiguous action potential firing patterns "regular spiking (RS)" and "repetitive burst spiking (RB)" (previously called intrinsically burst spiking) correlate well with a distinct morphology. RS pyramidal cells are somatodendritically of the slender-tufted type and possess numerous local intralaminar and intracolumnar axonal collaterals, mostly reaching layer I. By contrast, their transcolumnar projections are less well developed. The RB pyramidal cells are somatodendritically of the thick-tufted type and show only relatively sparse local axonal collaterals, which are preferentially emitted as long horizontal or oblique infragranular collaterals. However, contrary to many previous slice studies, a substantial number of these neurons also showed axonal collaterals reaching layer I. Thus, electrophysiologically defined pyramidal cells of layer Vb show an input and output pattern which suggests RS cells to be more "locally segregating" signal processors whereas RB cells seem to act more on a "global integrative" scale. PMID:27706253

  9. Erythropoietin administration alone or in combination with endurance training affects neither skeletal muscle morphology nor angiogenesis in healthy young men.

    PubMed

    Larsen, Mads S; Vissing, Kristian; Thams, Line; Sieljacks, Peter; Dalgas, Ulrik; Nellemann, Birgitte; Christensen, Britt

    2014-10-01

    The aim was to investigate the ability of an erythropoiesis-stimulating agent (ESA), alone or in combination with endurance training, to induce changes in human skeletal muscle fibre and vascular morphology. In a comparative study, 36 healthy untrained men were randomly dispersed into the following four groups: sedentary-placebo (SP, n = 9); sedentary-ESA (SE, n = 9); training-placebo (TP, n = 10); or training-ESA (TE, n = 8). The ESA or placebo was injected once weekly. Training consisted of progressive bicycling three times per week for 10 weeks. Before and after the intervention period, muscle biopsies and magnetic resonance images were collected from the thigh muscles, blood was collected, body composition measured and endurance exercise performance evaluated. The ESA treatment (SE and TE) led to elevated haematocrit, and both ESA treatment and training (SE, TP and TE) increased maximal O2 uptake. With regard to skeletal muscle morphology, TP alone exhibited increases in whole-muscle cross-sectional area and fibre diameter of all fibre types. Also exclusively for TP was an increase in type IIa fibres and a corresponding decrease in type IIx fibres. Furthermore, an overall training effect (TP and TE) was statistically demonstrated in whole-muscle cross-sectional area, muscle fibre diameter and type IIa and type IIx fibre distribution. With regard to muscle vascular morphology, TP and TE both promoted a rise in capillary to muscle fibre ratio, with no differences between the two groups. There were no effects of ESA treatment on any of the muscle morphological parameters. Despite the haematopoietic effects of ESA, we provide novel evidence that endurance training rather than ESA treatment induces adaptational changes in angiogenesis and muscle morphology.

  10. Quantitative and morphological changes of Langerhans cells in Bowen's disease from patients with chronic arsenicism.

    PubMed

    Wang, B J; Lee, Y Y; Mak, C P; Kao, H F; Hsu, M L; Hsien, J R

    1991-11-01

    Langerhans cells (LCs) are considered to be responsible for the immunologic presentation of tumor-associated antigens and play a role in the elimination of neoplastic clones. Ultraviolet light B can cause dysfunction and loss of LCs. Both the number and dendritic morphology of LCs are known to be diminished in squamous cell carcinomas from sun-exposed skin. The effects of arsenics on LCs are unknown. Using an OKT-6 monoclonal antibody to stain intraepithelial LCs, we compared their number and morphology in Bowen's lesions and in the perilesional skin from sun-protected sites in ten patients with chronic arsenicism. There was a significant reduction in the numbers of LCs in the Bowen's lesions as compared to the perilesional skin specimens. Loss of dendrites was observed in all Bowen's lesions and in seven of the perilesional skin specimens. Ultrastructurally, the LCs showed an absence of dendrites, but the Birbeck granules were preserved. Since the specimens were not from sun-exposed skin in our study, the findings may be related to chronic arsenic intoxication. The morphologic alteration of LCs observed in the perilesional skin further suggests an arsenic-related systemic dysfunction of the LCs, which in turn may contribute to the development of skin cancers in these patients.

  11. Morphological, electrophysiological, and synaptic properties of corticocallosal pyramidal cells in the neonatal rat neocortex.

    PubMed

    Le Bé, Jean-Vincent; Silberberg, Gilad; Wang, Yun; Markram, Henry

    2007-09-01

    Neocortical pyramidal cells (PCs) project to various cortical and subcortical targets. In layer V, the population of thick tufted PCs (TTCs) projects to subcortical targets such as the tectum, brainstem, and spinal cord. Another population of layer V PCs projects via the corpus callosum to the contralateral neocortical hemisphere mediating information transfer between the hemispheres. This subpopulation (corticocallosally projecting cells [CCPs]) has been previously described in terms of their morphological properties, but less is known about their electrophysiological properties, and their synaptic connectivity is unknown. We studied the morphological, electrophysiological, and synaptic properties of CCPs by retrograde labeling with fluorescent microbeads in P13-P16 Wistar rats. CCPs were characterized by shorter, untufted apical dendrites, which reached only up to layers II/III, confirming previous reports. Synaptic connections between CCPs were different from those observed between TTCs, both in probability of occurrence and dynamic properties. We found that the CCP network is about 4 times less interconnected than the TTC network and the probability of release is 24% smaller, resulting in a more linear synaptic transmission. The study shows that layer V pyramidal neurons projecting to different targets form subnetworks with specialized connectivity profiles, in addition to the specialized morphological and electrophysiological intrinsic properties.

  12. Stabilization of gene expression and cell morphology after explant recycling during fin explant culture in goldfish

    SciTech Connect

    Chenais, Nathalie; Lareyre, Jean-Jacques; Le Bail, Pierre-Yves; Labbe, Catherine

    2015-07-01

    The development of fin primary cell cultures for in vitro cellular and physiological studies is hampered by slow cell outgrowth, low proliferation rate, poor viability, and sparse cell characterization. Here, we investigated whether the recycling of fresh explants after a first conventional culture could improve physiological stability and sustainability of the culture. The recycled explants were able to give a supplementary cell culture showing faster outgrowth, cleaner cell layers and higher net cell production. The cells exhibited a highly stabilized profile for marker gene expression including a low cytokeratin 49 (epithelial marker) and a high collagen 1a1 (mesenchymal marker) expression. Added to the cell spindle-shaped morphology, motility behavior, and actin organization, this suggests that the cells bore stable mesenchymal characteristics. This contrast with the time-evolving expression pattern observed in the control fresh explants during the first 2 weeks of culture: a sharp decrease in cytokeratin 49 expression was concomitant with a gradual increase in col1a1. We surmise that such loss of epithelial features for the benefit of mesenchymal ones was triggered by an epithelial to mesenchymal transition (EMT) process or by way of a progressive population replacement process. Overall, our findings provide a comprehensive characterization of this new primary culture model bearing mesenchymal features and whose stability over culture time makes those cells good candidates for cell reprogramming prior to nuclear transfer, in a context of fish genome preservation. - Highlights: • Recycled fin explants outgrow cells bearing stable mesenchymal traits. • Cell production and quality is enhanced in the recycled explant culture system. • Fresh fin primary culture is highly variable and loose epithelial traits over time.

  13. Primary cilia mechanics affects cell mechanosensation: A computational study.

    PubMed

    Khayyeri, Hanifeh; Barreto, Sara; Lacroix, Damien

    2015-08-21

    Primary cilia (PC) are mechanical cell structures linked to the cytoskeleton and are central to how cells sense biomechanical signals from their environment. However, it is unclear exactly how PC mechanics influences cell mechanosensation. In this study we investigate how the PC mechanical characteristics are involved in the mechanotransduction process whereby cilium deflection under fluid flow induces strains on the internal cell components that regulate the cell׳s mechanosensitive response. Our investigation employs a computational approach in which a finite element model of a cell consisting of a nucleus, cytoplasm, cortex, microtubules, actin bundles and a primary cilium was used together with a finite element representation of a flow chamber. Fluid-structure interaction analysis was performed by simulating perfusion flow of 1mm/s on the cell model. Simulations of cells with different PC mechanical characteristics, showed that the length and the stiffness of PC are responsible for the transmission of mechanical stimuli to the cytoskeleton. Fluid flow deflects the cilium, with the highest strains found at the base of the PC and in the cytoplasm. The PC deflection created further strains on the cell nucleus but did not influence microtubules and actin bundles significantly. Our results indicate that PC deflection under fluid flow stimulation transmits mechanical strain primarily to other essential organelles in the cytoplasm, such as the Golgi complex, that regulate cells' mechanoresponse. The simulations further suggest that cell mechanosensitivity can be altered by targeting PC length and rigidity.

  14. Morphological interactions of interdigitating dendritic cells with B and T cells in human mesenteric lymph nodes.

    PubMed

    Takahashi, K; Kenji, A; Norihiro, T; Eisaku, K; Takashi, O; Kazuhiko, H; Tadashi, Y; Tadaatsu, A

    2001-07-01

    Interdigitating dendritic cells (IDC) of the human mesenteric lymph nodes (LN) were examined by two-color immunofluorescent microscopy and flow cytometry to clarify their exact localization, immunophenotype, and relationships with T and B cells. IDC were identified as HLA-DR(bright) large dendriform cells of the T cell areas co-expressing CD40, CD54 (ICAM-1), CD80 (B7/B7-1), CD83, and CD86 (B70/B7-2). The majority of IDC directly attached to a few IgD+ naive B cells as well as to numerous CD4+ T cells. When LN cells were singly suspended and briefly incubated in vitro, IDC formed clusters with IgD+ IgM+ naive B cells, but not with IgA+ or IgG+ B cells. When suspended LN cells were cultured, clustered B cells disappeared within 7 days, and on prolonged culture, some IDC developed into extensively dendriform cells forming stable complexes with several or sometimes numerous CD4+ IL-2R+ CD40L+ activated T cells. These findings indicate that resting naive B cells actually interact with IDC directly in T cell areas of human secondary lymphoid tissues. IDC have a non-antigen (Ag)-specific, strong affinity for resting naive B cells, but this affinity is transient and IDC cannot form stable complexes with B cells, although they can form stable complexes with activated T cells. It is suggested that the stable IDC/Ag-activated T cell complexes make it possible to capture and to stimulate rare Ag-specific resting naive B cells with high efficiency.

  15. ETM study of electroporation influence on cell morphology in human malignant melanoma and human primary gingival fibroblast cells

    PubMed Central

    Skolucka, Nina; Daczewska, Malgorzata; Saczko, Jolanta; Chwilkowska, Agnieszka; Choromanska, Anna; Kotulska, Malgorzata; Kaminska, Iwona; Kulbacka, Julita

    2011-01-01

    Objective To estimate electroporation (EP) influence on malignant and normal cells. Methods Two cell lines including human malignant melanoma (Me-45) and normal human gingival fibroblast (HGFs) were used. EP parameters were the following: 250, 1 000, 1 750, 2 500 V/cm; 50 µs by 5 impulses for every case. The viability of cells after EP was estimated by MTT assay. The ultrastructural analysis was observed by transmission electron microscope (Zeiss EM 900). Results In the current study we observed the intracellular effect following EP on Me-45 and HGF cells. At the conditions applied, we did not observe any significant damage of mitochondrial activity in both cell lines treated by EP. Conversely, we showed that EP in some conditions can stimulate cells to proliferation. Some changes induced by EP were only visible in electron microscopy. In fibroblast cells we observed significant changes in lower parameters of EP (250 and 1 000 V/cm). After applying higher electric field intensities (2 500 V/cm) we detected many vacuoles, myelin-like bodies and swallowed endoplasmic reticulum. In melanoma cells such strong pathological modifications after EP were not observed, in comparison with control cells. The ultrastructure of both treated cell lines was changed according to the applied parameters of EP. Conclusions We can claim that EP conditions are cell line dependent. In terms of the intracellular morphology, human fibroblasts are more sensitive to electric field as compared with melanoma cells. Optimal conditions should be determined for each cell line. Summarizing our study, we can conclude that EP is not an invasive method for human normal and malignant cells. This technique can be safely applied in chemotherapy for delivering drugs into tumor cells. PMID:23569735

  16. Morphology and behaviour of dinoflagellate chromosomes during the cell cycle and mitosis.

    PubMed

    Bhaud, Y; Guillebault, D; Lennon, J; Defacque, H; Soyer-Gobillard, M O; Moreau, H

    2000-04-01

    The morphology and behaviour of the chromosomes of dinoflagellates during the cell cycle appear to be unique among eukaryotes. We used synchronized and aphidicolin-blocked cultures of the dinoflagellate Crypthecodinium cohnii to describe the successive morphological changes that chromosomes undergo during the cell cycle. The chromosomes in early G(1) phase appeared to be loosely condensed with numerous structures protruding toward the nucleoplasm. They condensed in late G(1), before unwinding in S phase. The chromosomes in cells in G(2) phase were tightly condensed and had a double number of arches, as visualised by electron microscopy. During prophase, chromosomes elongated and split longitudinally, into characteristic V or Y shapes. We also used confocal microscopy to show a metaphase-like alignment of the chromosomes, which has never been described in dinoflagellates. The metaphase-like nucleus appeared flattened and enlarged, and continued to do so into anaphase. Chromosome segregation occurred via binding to the nuclear envelope surrounding the cytoplasmic channels and microtubule bundles. Our findings are summarized in a model of chromosome behaviour during the cell cycle.

  17. Dendritic Morphology of Caudal Periaqueductal Gray Projecting Retinal Ganglion Cells in Mongolian Gerbil (Meriones unguiculatus)

    PubMed Central

    Ren, Chaoran; Pu, Mingliang; Cui, Qi; So, Kwok-Fai

    2014-01-01

    In this study we investigated the morphological features of the caudal periaqueductal gray (cPAG)-projecting retinal ganglion cells (RGCs) in Mongolian gerbils using retrograde labeling, in vitro intracellular injection, confocal microscopy and three-dimensional reconstruction approaches. cPAG-projecting RGCs exhibit small somata (10–17 µm) and irregular dendritic fields (201–298 µm). Sizes of somata and dendritic fields do not show obvious variation at different distance from the optic disk (eccentricity). Dendrites are moderately branched. Morphological analysis (n = 23) reveals that cPAG-projecting RGCs ramified in sublamina a and b in the inner plexiform layer. These cells exhibit different stratification patterns based on the thickness of dendritic bands in sublaminas a and b: majority of analyzed cells (16 out of 23) have two bands of arborizations share similar thickness. The rest of analyzed cells (7 out of 23) exhibit thinner band in sublamina a than in sublamina b. Together, the present study suggests that cPAG of Mongolian gerbil could receive direct retinal inputs from two types of bistratified RGCs. Furthermore, a small subset of melanopsin-expressing RGCs (total 41 in 6 animals) is shown to innervate the rostral PAG (rPAG). Functional characteristics of these non-visual center projecting RGCs remain to be determined. PMID:25054882

  18. Morphology studies on high-temperature polymer electrolyte membrane fuel cell electrodes

    NASA Astrophysics Data System (ADS)

    Mack, Florian; Klages, Merle; Scholta, Joachim; Jörissen, Ludwig; Morawietz, Tobias; Hiesgen, Renate; Kramer, Dominik; Zeis, Roswitha

    2014-06-01

    The electrode morphology influences the properties and performance of polymer electrolyte membrane fuel cells (PEMFC). Here we report our studies of two different electrodes for high-temperature PEMFC prepared by spraying and coating and their impact on the fuel cell performance. Differences in 3D microstructure and adhesion between catalyst layer and gas diffusion layer (GDL) of the electrodes were studied with X-ray microtomography. Scanning electrode microscope investigations show hairline cracks between agglomerates on the surface of the sprayed electrode, whereas the coated electrode shows a network of shrinkage cracks in the catalyst layer. The distribution of the electrode binder polytetrafluoroethylene (PTFE) is related to the locally resolved conductivity, which was determined by scanning the electrode surfaces with a conductive atomic force microscopy (AFM) tip. The macrostructures of the sprayed and coated electrodes are different but contain similar pore structures. The coated electrode has a higher PTFE concentration on the top region, which tends to form a nonconductive and less wettable "skin" on the electrode surface and delays the start-up of the fuel cell. In contrast to low-temperature PEMFC, the electrode morphology has only a minor impact on the steady-state cell performance of high-temperature PEMFC.

  19. Neogenin and RGMa control neural tube closure and neuroepithelial morphology by regulating cell polarity.

    PubMed

    Kee, Nigel; Wilson, Nicole; De Vries, Melissa; Bradford, DanaKai; Key, Brian; Cooper, Helen M

    2008-11-26

    In humans, neural tube closure defects occur in 1:1000 pregnancies. The design of new strategies for the prevention of such common defects would benefit from an improved understanding of the molecular events underlying neurulation. Neural fold elevation is a key morphological process that acts during neurulation to drive neural tube closure. However, to date, the molecular pathways underpinning neural fold elevation have not been elucidated. Here, we use morpholino knock-down technology to demonstrate that Repulsive Guidance Molecule (RGMa)-Neogenin interactions are essential for effective neural fold elevation during Xenopus neurulation and that loss of these molecules results in disrupted neural tube closure. We demonstrate that Neogenin and RGMa are required for establishing the morphology of deep layer cells in the neural plate throughout neurulation. We also show that loss of Neogenin severely disrupts the microtubule network within the deep layer cells suggesting that Neogenin-dependent microtubule organization within the deep cells is essential for radial intercalation with the overlying superficial cell layer, thereby driving neural fold elevation. In addition, we show that sustained Neogenin activity is also necessary for the establishment of the apicobasally polarized pseudostratified neuroepithelium of the neural tube. Therefore, our study identifies a novel signaling pathway essential for radial intercalation and epithelialization during neural fold elevation and neural tube morphogenesis.

  20. Acquired cystic disease-associated renal cell carcinoma: further characterization of the morphologic and immunopathologic features.

    PubMed

    Ahn, Soomin; Kwon, Ghee Young; Cho, Yong Mee; Jun, Sun-Young; Choi, Chan; Kim, Hyun-Jung; Park, Yong Wook; Park, Weon Seo; Shim, Jung Won

    2013-12-01

    Acquired cystic disease-associated renal cell carcinoma (ACD-RCC) is a subtype of renal cell carcinoma (RCC) with unique morphologic features found exclusively in the background of end-stage renal disease. We analyzed the clinicopathologic features and immumoreactive profiles of 12 cases of ACD-RCC to further characterize this recently recognized entity. Review of histologic slides was performed in conjunction with immunohistochemical staining directed to the contemporary diagnostic antibodies and the putative target therapy-related markers. Histologically, the tumors showed characteristic inter-or intracellular microlumens and eosinophilic tumor cells. Intratumoral hemosiderin deposition and degenerating foamy tumor cells were consistent findings which were not previously described. Immunohistochemically, all the tumors were positive for alpha-methylacyl-CoA-racemase, CD10, pan-cytokeratin, PTEN (phosphatase and tensin homolog deleted on chromosome 10) and c-met, while negative for carbonic anhydrase-9, CD57, CD68, c-kit, pax-2, platelet-derived growth factor receptor (PDGFR)-α or vascular endothelial growth factor receptor (VEGFR)-2. Heterogenous staining was found for CK7 and kidney-specific cadherin. Positive reaction to c-met suggests its utility as a plausible therapeutic target in ACD-RCC. Thus, we present the unique morphologic and immunopathologic features of ACD-RCC, which may be helpful in both diagnostic and therapeutic aspects. PMID:23471757

  1. Epithelial cell morphology and adhesion on diamond films deposited and chemically modified by plasma processes.

    PubMed

    Rezek, Bohuslav; Ukraintsev, Egor; Krátká, Marie; Taylor, Andrew; Fendrych, Frantisek; Mandys, Vaclav

    2014-09-01

    The authors show that nanocrystalline diamond (NCD) thin films prepared by microwave plasma enhanced chemical vapor deposition apparatus with a linear antenna delivery system are well compatible with epithelial cells (5637 human bladder carcinoma) and significantly improve the cell adhesion compared to reference glass substrates. This is attributed to better adhesion of adsorbed layers to diamond as observed by atomic force microscopy (AFM) beneath the cells. Moreover, the cell morphology can be adjusted by appropriate surface treatment of diamond by using hydrogen and oxygen plasma. Cell bodies, cytoplasmic rims, and filopodia were characterized by Peakforce AFM. Oxidized NCD films perform better than other substrates under all conditions (96% of cells adhered well). A thin adsorbed layer formed from culture medium and supplemented with fetal bovine serum (FBS) covered the diamond surface and played an important role in the cell adhesion. Nevertheless, 50-100 nm large aggregates formed from the RPMI medium without FBS facilitated cell adhesion also on hydrophobic hydrogenated NCD (increase from 23% to 61%). The authors discuss applicability for biomedical uses.

  2. Morphology and dynamic scaling analysis of cell colonies with linear growth fronts

    NASA Astrophysics Data System (ADS)

    Huergo, M. A. C.; Pasquale, M. A.; Bolzán, A. E.; Arvia, A. J.; González, P. H.

    2010-09-01

    The growth of linear cell colony fronts is investigated from the morphology of cell monolayer colonies, the cell size and shape distribution, the front displacement velocity, and the dynamic scaling analysis of front roughness fluctuations. At the early growth stages, colony patterns consist of rather ordered compact domains of small cells, whereas at advanced stages, an uneven distribution of cells sets in, and some large cells and cells exhibiting large filopodia are produced. Colony front profiles exhibit overhangs and behave as fractals with the dimension DF=1.25±0.05 . The colony fronts shift at 0.22±0.02μmmin-1 average constant linear velocity and their roughness (w) increases with time (t) . Dynamic scaling analysis of experimental and overhang-corrected growth profile data shows that w versus system width l log-log plots collapse to a single curve when l exceeds a certain threshold value lo , a width corresponding to the average diameter of few cells. Then, the influence of overhangs on the roughness dynamics becomes negligible, and a growth exponent β=0.33±0.02 is derived. From the structure factor analysis of overhang-corrected profiles, a global roughness exponent αs=0.50±0.05 is obtained. For l>200μm , this set of exponents fulfills the Family-Vicsek relationship. It is consistent with the predictions of the continuous Kardar-Parisi-Zhang model.

  3. Morphological control of hybrid polymer-quantum dot solar cells with electron acceptor ligands

    NASA Astrophysics Data System (ADS)

    Boivin, Mathieu; Lamarre, Sébastien; Tessier, Jonathan; Lecavalier, Marie-Ève; Najari, Ahmed; Dufour-Beauséjour, Sophie; Brown Dussault, Evelyne; Collin, Pierre; Allen, Claudine Nı.

    2012-01-01

    We integrate the electro-attractive conjugated molecule tetrafluoro-tetracyano-quinodimethane (F4TCNQ) in the active layer of polymer-CdSe colloidal quantum dot (cQD) solar cells. The addition of this molecule enhances cQD dispersion inside the polymer. In tuning its concentration, we can optimize the active layer morphology for charge separation and transport. A smoother morphology is likely the result of polymer chain adsorption on cQDs via F4TCNQ which increases the steric barrier between cQDs. Our most optimized device has a F4TCNQ:cQDs weight ratio of 0.5% improving the power conversion efficiency by a factor ˜2.3.

  4. Physiological and morphological characterization of ganglion cells in the salamander retina.

    PubMed

    Wang, Jing; Jacoby, Roy; Wu, Samuel M

    2016-02-01

    Retinal ganglion cells (RGCs) integrate visual information from the retina and transmit collective signals to the brain. A systematic investigation of functional and morphological characteristics of various types of RGCs is important to comprehensively understand how the visual system encodes and transmits information via various RGC pathways. This study evaluated both physiological and morphological properties of 67 RGCs in dark-adapted flat-mounted salamander retina by examining light-evoked cation and chloride current responses via voltage-clamp recordings and visualizing morphology by Lucifer yellow fluorescence with a confocal microscope. Six groups of RGCs were described: asymmetrical ON-OFF RGCs, symmetrical ON RGCs, OFF RGCs, and narrow-, medium- and wide-field ON-OFF RGCs. Dendritic field diameters of RGCs ranged 102-490 μm: narrow field (<200 μm, 31% of RGCs), medium field (200-300 μm, 45%) and wide field (>300 μm, 24%). Dendritic ramification patterns of RGCs agree with the sublamina A/B rule. 34% of RGCs were monostratified, 24% bistratified and 42% diffusely stratified. 70% of ON RGCs and OFF RGCs were monostratified. Wide-field RGCs were diffusely stratified. 82% of RGCs generated light-evoked ON-OFF responses, while 11% generated ON responses and 7% OFF responses. Response sensitivity analysis suggested that some RGCs obtained separated rod/cone bipolar cell inputs whereas others obtained mixed bipolar cell inputs. 25% of neurons in the RGC layer were displaced amacrine cells. Although more types may be defined by more refined classification criteria, this report is to incorporate more physiological properties into RGC classification.

  5. Physiological and morphological characterization of ganglion cells in the salamander retina.

    PubMed

    Wang, Jing; Jacoby, Roy; Wu, Samuel M

    2016-02-01

    Retinal ganglion cells (RGCs) integrate visual information from the retina and transmit collective signals to the brain. A systematic investigation of functional and morphological characteristics of various types of RGCs is important to comprehensively understand how the visual system encodes and transmits information via various RGC pathways. This study evaluated both physiological and morphological properties of 67 RGCs in dark-adapted flat-mounted salamander retina by examining light-evoked cation and chloride current responses via voltage-clamp recordings and visualizing morphology by Lucifer yellow fluorescence with a confocal microscope. Six groups of RGCs were described: asymmetrical ON-OFF RGCs, symmetrical ON RGCs, OFF RGCs, and narrow-, medium- and wide-field ON-OFF RGCs. Dendritic field diameters of RGCs ranged 102-490 μm: narrow field (<200 μm, 31% of RGCs), medium field (200-300 μm, 45%) and wide field (>300 μm, 24%). Dendritic ramification patterns of RGCs agree with the sublamina A/B rule. 34% of RGCs were monostratified, 24% bistratified and 42% diffusely stratified. 70% of ON RGCs and OFF RGCs were monostratified. Wide-field RGCs were diffusely stratified. 82% of RGCs generated light-evoked ON-OFF responses, while 11% generated ON responses and 7% OFF responses. Response sensitivity analysis suggested that some RGCs obtained separated rod/cone bipolar cell inputs whereas others obtained mixed bipolar cell inputs. 25% of neurons in the RGC layer were displaced amacrine cells. Although more types may be defined by more refined classification criteria, this report is to incorporate more physiological properties into RGC classification. PMID:26731645

  6. Basal cell adenocarcinoma and Basal cell adenoma of the salivary glands: a clinicopathological review of seventy tumors with comparison of morphologic features and growth control indices.

    PubMed

    Wilson, Thomas C; Robinson, Robert A

    2015-06-01

    Basal cell adenoma and basal cell adenocarcinoma represent uncommon basaloid salivary gland neoplasms that show marked morphologic similarity. We wished to compare clinical outcome and morphologic features as well as growth and proliferation associated markers for both neoplasms. We reviewed the pathologic features of 70 neoplasms diagnosed as basal cell adenoma or basal cell adenocarcinoma. Observations included maximum mitotic activity and presence or absence of invasion into surrounding normal tissues as well as immunohistochemical studies for Ki-67, caspase 3, p53, and bcl-2. Establishing malignancy on the basis of invasion into surrounding benign tissues, 41 basal cell adenomas and 29 basal cell adenocarcinomas were identified. For tumors with follow-up, recurrence rates were 6.7 % for basal cell adenoma and 16.7 % for basal cell adenocarcinoma. One patient with basal cell adenocarcinoma had distant metastases and died of disease. Overall basal cell adenocarcinomas showed significantly higher values for growth and proliferation markers compared to basal cell adenomas. Salivary gland basal cell adenoma and basal cell adenocarcinoma show morphologic similarity. Basal cell adenocarcinoma can exhibit a locally aggressive behavior and has potential metastatic behavior. The overall mitotic rate and Ki-67 expression were higher in basal cell adenocarcinoma compared to basal cell adenoma, but overlap between the results of these observations in each tumor did not allow for accurate diagnosis or prediction of outcome in individual cases. We conclude that morphologic observation of local tissue invasion is the best marker for separating basal cell adenoma from basal cell adenocarcinoma.

  7. Plastic hatching timing by red-eyed treefrog embryos interacts with larval predator identity and sublethal predation to affect prey morphology but not performance.

    PubMed

    Touchon, Justin C; Wojdak, Jeremy M

    2014-01-01

    Many animals respond to predation risk by altering their morphology, behavior, or life-history. We know a great deal about the cues prey respond to and the changes to prey that can be induced by predation risk, but less is known about how plastic responses to predators may be affected by separate plastic responses occurring earlier in life, particularly during the embryonic period. Embryos of a broad array of taxa can respond to egg- or larval-stage risks by altering hatching timing, which may alter the way organisms respond to future predators. Using the red-eyed treefrog (Agalychnis callidryas), a model for understanding the effects of plasticity across life-stages, we assessed how the combined effects of induced variation in the timing of embryo hatching and variation in the larval predator community impacted tadpole morphology, pigmentation and swimming performance. We found that A. callidryas tadpoles developed deeper tail muscles and fins and darker pigmentation in response to fish predators, either when alone or in diverse community with other predators. Tadpoles altered morphology much less so to dragonfly naiads or water bugs. Interestingly, morphological responses to predators were also affected by induced differences in hatching age, with early and late-hatched tadpoles exhibiting different allometric relationships between tail height and body length in different predator environments. Beyond induced morphological changes, fish predators often damaged tadpoles' tails without killing them (i.e., sublethal predation), but these tadpoles swam equally quickly to those with fully intact tails. This was due to the fact that tadpoles with more damaged tails increased tail beats to achieve equal swimming speed. This study demonstrates that plastic phenotypic responses to predation risk can be influenced by a complex combination of responses to both the embryo and larval environments, but also that prey performance can be highly resilient to sublethal predation.

  8. Plastic hatching timing by red-eyed treefrog embryos interacts with larval predator identity and sublethal predation to affect prey morphology but not performance.

    PubMed

    Touchon, Justin C; Wojdak, Jeremy M

    2014-01-01

    Many animals respond to predation risk by altering their morphology, behavior, or life-history. We know a great deal about the cues prey respond to and the changes to prey that can be induced by predation risk, but less is known about how plastic responses to predators may be affected by separate plastic responses occurring earlier in life, particularly during the embryonic period. Embryos of a broad array of taxa can respond to egg- or larval-stage risks by altering hatching timing, which may alter the way organisms respond to future predators. Using the red-eyed treefrog (Agalychnis callidryas), a model for understanding the effects of plasticity across life-stages, we assessed how the combined effects of induced variation in the timing of embryo hatching and variation in the larval predator community impacted tadpole morphology, pigmentation and swimming performance. We found that A. callidryas tadpoles developed deeper tail muscles and fins and darker pigmentation in response to fish predators, either when alone or in diverse community with other predators. Tadpoles altered morphology much less so to dragonfly naiads or water bugs. Interestingly, morphological responses to predators were also affected by induced differences in hatching age, with early and late-hatched tadpoles exhibiting different allometric relationships between tail height and body length in different predator environments. Beyond induced morphological changes, fish predators often damaged tadpoles' tails without killing them (i.e., sublethal predation), but these tadpoles swam equally quickly to those with fully intact tails. This was due to the fact that tadpoles with more damaged tails increased tail beats to achieve equal swimming speed. This study demonstrates that plastic phenotypic responses to predation risk can be influenced by a complex combination of responses to both the embryo and larval environments, but also that prey performance can be highly resilient to sublethal predation

  9. Plastic Hatching Timing by Red-Eyed Treefrog Embryos Interacts with Larval Predator Identity and Sublethal Predation to Affect Prey Morphology but Not Performance

    PubMed Central

    Touchon, Justin C.; Wojdak, Jeremy M.

    2014-01-01

    Many animals respond to predation risk by altering their morphology, behavior, or life-history. We know a great deal about the cues prey respond to and the changes to prey that can be induced by predation risk, but less is known about how plastic responses to predators may be affected by separate plastic responses occurring earlier in life, particularly during the embryonic period. Embryos of a broad array of taxa can respond to egg- or larval-stage risks by altering hatching timing, which may alter the way organisms respond to future predators. Using the red-eyed treefrog (Agalychnis callidryas), a model for understanding the effects of plasticity across life-stages, we assessed how the combined effects of induced variation in the timing of embryo hatching and variation in the larval predator community impacted tadpole morphology, pigmentation and swimming performance. We found that A. callidryas tadpoles developed deeper tail muscles and fins and darker pigmentation in response to fish predators, either when alone or in diverse community with other predators. Tadpoles altered morphology much less so to dragonfly naiads or water bugs. Interestingly, morphological responses to predators were also affected by induced differences in hatching age, with early and late-hatched tadpoles exhibiting different allometric relationships between tail height and body length in different predator environments. Beyond induced morphological changes, fish predators often damaged tadpoles’ tails without killing them (i.e., sublethal predation), but these tadpoles swam equally quickly to those with fully intact tails. This was due to the fact that tadpoles with more damaged tails increased tail beats to achieve equal swimming speed. This study demonstrates that plastic phenotypic responses to predation risk can be influenced by a complex combination of responses to both the embryo and larval environments, but also that prey performance can be highly resilient to sublethal predation

  10. Piper and Vismia species from Colombian Amazonia differentially affect cell proliferation of hepatocarcinoma cells.

    PubMed

    Lizcano, Leandro J; Siles, Maite; Trepiana, Jenifer; Hernández, M Luisa; Navarro, Rosaura; Ruiz-Larrea, M Begoña; Ruiz-Sanz, José Ignacio

    2015-01-01

    There is an increasing interest to identify plant-derived natural products with antitumor activities. In this work, we have studied the effects of aqueous leaf extracts from Amazonian Vismia and Piper species on human hepatocarcinoma cell toxicity. Results showed that, depending on the cell type, the plants displayed differential effects; thus, Vismia baccifera induced the selective killing of HepG2, while increasing cell growth of PLC-PRF and SK-HEP-1. In contrast, these two last cell lines were sensitive to the toxicity by Piper krukoffii and Piper putumayoense, while the Piperaceae did not affect HepG2 growth. All the extracts induced cytotoxicity to rat hepatoma McA-RH7777, but were innocuous (V. baccifera at concentrations < 75 µg/mL) or even protected cells from basal death (P. putumayoense) in primary cultures of rat hepatocytes. In every case, cytotoxicity was accompanied by an intracellular accumulation of reactive oxygen species (ROS). These results provide evidence for the anticancer activities of the studied plants on specific cell lines and suggest that cell killing could be mediated by ROS, thus involving mechanisms independent of the plants free radical scavenging activities. Results also support the use of these extracts of the Vismia and Piper genera with opposite effects as a model system to study the mechanisms of the antitumoral activity against different types of hepatocarcinoma. PMID:25558904

  11. Changes in lipid metabolism and cell morphology following attack by phospholipase C (Clostridium perfringens) on red cells or lymphocytes.

    PubMed

    Allan, D; Low, M G; Finean, J B; Michell, R H

    1975-12-01

    When intact human erythrocytes were treated with phospholipase C (Clostridium perfringens), up to 30% of the membrane phospholipids were broken down without significant cell lysis. Only phosphatidylcholine and sphingomyelin were attacked. Ceramide (derived from sphingomyelin) accumulated, but 1,2-diacylglycerol (derived from phosphatidylcholine) was largely converted into phosphatidate. Up to 12% of the cell phospholipid could be converted into phosphatidate in this way. Pig erythrocytes and lymphocytes showed a similar but smaller synthesis of phosphatidate after phospholipase C attack. Phospholipase C also caused a marked morphological change in erythrocytes, giving rise to spherical cells containing internal membrane vesicles. This change appeared to be due to ceramide and de and diacylglycerol accumulation rather than to increased phosphatidate content of the cells.

  12. How does metabolism affect cell death in cancer?

    PubMed

    Villa, Elodie; Ricci, Jean-Ehrland

    2016-07-01

    In cancer research, identifying a specificity of tumor cells compared with 'normal' proliferating cells for targeted therapy is often considered the Holy Grail for researchers and clinicians. Although diverse in origin, most cancer cells share characteristics including the ability to escape cell death mechanisms and the utilization of different methods of energy production. In the current paradigm, aerobic glycolysis is considered the central metabolic characteristic of cancer cells (Warburg effect). However, recent data indicate that cancer cells also show significant changes in other metabolic pathways. Indeed, it was recently suggested that Kreb's cycle, pentose phosphate pathway intermediates, and essential and nonessential amino acids have key roles. Renewed interest in the fact that cancer cells have to reprogram their metabolism in order to proliferate or resist treatment must take into consideration the ability of tumor cells to adapt their metabolism to the local microenvironment (low oxygen, low nutrients). This variety of metabolic sources might be either a strength, resulting in infinite possibilities for adaptation and increased ability to resist chemotherapy-induced death, or a weakness that could be targeted to kill cancer cells. Here, we discuss recent insights showing how energetic metabolism may regulate cell death and how this might be relevant for cancer treatment.

  13. Stem cell origin differently affects bone tissue engineering strategies

    PubMed Central

    Mattioli-Belmonte, Monica; Teti, Gabriella; Salvatore, Viviana; Focaroli, Stefano; Orciani, Monia; Dicarlo, Manuela; Fini, Milena; Orsini, Giovanna; Di Primio, Roberto; Falconi, Mirella

    2015-01-01

    Bone tissue engineering approaches are encouraging for the improvement of conventional bone grafting technique drawbacks. Thanks to their self-renewal and multi-lineage differentiation ability, stem cells are one of the major actors in tissue engineering approaches, and among these adult mesenchymal stem cells (MSCs) hold a great promise for regenerative medicine strategies. Bone marrow MSCs (BM-MSCs) are the first- identified and well-recognized stem cell population used in bone tissue engineering. Nevertheless, several factors hamper BM-MSC clinical application and subsequently, new stem cell sources have been investigated for these purposes. The fruitful selection and combination of tissue engineered scaffold, progenitor cells, and physiologic signaling molecules allowed the surgeon to reconstruct the missing natural tissue. On the basis of these considerations, we analyzed the capability of two different scaffolds, planned for osteochondral tissue regeneration, to modulate differentiation of adult stem cells of dissimilar local sources (i.e., periodontal ligament, maxillary periosteum) as well as adipose-derived stem cells (ASCs), in view of possible craniofacial tissue engineering strategies. We demonstrated that cells are differently committed toward the osteoblastic phenotype and therefore, taking into account their specific features, they could be intriguing cell sources in different stem cell-based bone/periodontal tissue regeneration approaches. PMID:26441682

  14. Single-cell mass spectrometry reveals small molecules that affect cell fates in the 16-cell embryo

    PubMed Central

    Onjiko, Rosemary M.; Moody, Sally A.; Nemes, Peter

    2015-01-01

    Spatial and temporal changes in molecular expression are essential to embryonic development, and their characterization is critical to understand mechanisms by which cells acquire different phenotypes. Although technological advances have made it possible to quantify expression of large molecules during embryogenesis, little information is available on metabolites, the ultimate indicator of physiological activity of the cell. Here, we demonstrate that single-cell capillary electrophoresis-electrospray ionization mass spectrometry is able to test whether differential expression of the genome translates to the domain of metabolites between single embryonic cells. Dissection of three different cell types with distinct tissue fates from 16-cell embryos of the South African clawed frog (Xenopus laevis) and microextraction of their metabolomes enabled the identification of 40 metabolites that anchored interconnected central metabolic networks. Relative quantitation revealed that several metabolites were differentially active between the cell types in the wild-type, unperturbed embryos. Altering postfertilization cytoplasmic movements that perturb dorsal development confirmed that these three cells have characteristic small-molecular activity already at cleavage stages as a result of cell type and not differences in pigmentation, yolk content, cell size, or position in the embryo. Changing the metabolite concentration caused changes in cell movements at gastrulation that also altered the tissue fates of these cells, demonstrating that the metabolome affects cell phenotypes in the embryo. PMID:25941375

  15. Effect of ZnO morphology on affecting bactericidal property of ultra high molecular weight polyethylene biocomposite.

    PubMed

    Sharma, Rajeev Kumar; Agarwal, Meenakshi; Balani, Kantesh

    2016-05-01

    Bacterial infection of implants can be controlled by selective trapping of bacteria, followed with consequent killing by targeted antibacterial agents. Herein, the role of various ZnO morphologies, viz. micro-rods (R), nanoparticles (NP), and micro-disks (D) on antibacterial efficacy of ZnO via release of Zn(2+) and H2O2 is assessed, both as isolated powders and via incorporating them in cytocompatible ultra high molecular weight polyethylene (UHMWPE). Though ZnO is antibacterial, interestingly, all ZnO morphologies elicited a supportive growth of gram-negative bacteria (Escherichia coli) in culture medium (until 28-35 μg/ml). But, all ZnO morphologies did elicit bactericidal effect on gram positive bacteria (Staphylococcus aureus or Staphylococcus epidermidis) both in culture medium (for 0-2.5 μg/ml) or when incorporated (5-20 wt.%) into UHMWPE. The bactericidal mechanisms were quantified for various ZnO morphologies via: (i) H2O2 production, (ii) Zn(2+) release, and (iii) the presence of surface oxygen vacancies. On one hand, where only ZnO(NP) elicited release of H2O2 in the absence of light, maximum Zn(2+) release was elicited by ZnO(D). Interestingly, when ZnO is incorporated as reinforcement (5-20 wt.%), its antibacterial action against E. coli was vividly observed due to selective proliferation of bacteria only on friendly UHMWPE matrix. Hence, luring bacteria on affable UHMWPE surface can be complemented with their targeted killing by ZnO present in composite.

  16. Effect of ZnO morphology on affecting bactericidal property of ultra high molecular weight polyethylene biocomposite.

    PubMed

    Sharma, Rajeev Kumar; Agarwal, Meenakshi; Balani, Kantesh

    2016-05-01

    Bacterial infection of implants can be controlled by selective trapping of bacteria, followed with consequent killing by targeted antibacterial agents. Herein, the role of various ZnO morphologies, viz. micro-rods (R), nanoparticles (NP), and micro-disks (D) on antibacterial efficacy of ZnO via release of Zn(2+) and H2O2 is assessed, both as isolated powders and via incorporating them in cytocompatible ultra high molecular weight polyethylene (UHMWPE). Though ZnO is antibacterial, interestingly, all ZnO morphologies elicited a supportive growth of gram-negative bacteria (Escherichia coli) in culture medium (until 28-35 μg/ml). But, all ZnO morphologies did elicit bactericidal effect on gram positive bacteria (Staphylococcus aureus or Staphylococcus epidermidis) both in culture medium (for 0-2.5 μg/ml) or when incorporated (5-20 wt.%) into UHMWPE. The bactericidal mechanisms were quantified for various ZnO morphologies via: (i) H2O2 production, (ii) Zn(2+) release, and (iii) the presence of surface oxygen vacancies. On one hand, where only ZnO(NP) elicited release of H2O2 in the absence of light, maximum Zn(2+) release was elicited by ZnO(D). Interestingly, when ZnO is incorporated as reinforcement (5-20 wt.%), its antibacterial action against E. coli was vividly observed due to selective proliferation of bacteria only on friendly UHMWPE matrix. Hence, luring bacteria on affable UHMWPE surface can be complemented with their targeted killing by ZnO present in composite. PMID:26952491

  17. Pseudomonas aeruginosa N-3-oxo-dodecanoyl-homoserine Lactone Elicits Changes in Cell Volume, Morphology, and AQP9 Characteristics in Macrophages.

    PubMed

    Holm, Angelika; Magnusson, Karl-Eric; Vikström, Elena

    2016-01-01

    Quorum sensing (QS) communication allows Pseudomonas aeruginosa to collectively control its population density and the production of biofilms and virulence factors. QS signal molecules, like N-3-oxo-dodecanoyl-L-homoserine lactone (3O-C12-HSL), can also affect the behavior of host cells, e.g., by modulating the chemotaxis, migration, and phagocytosis of human leukocytes. Moreover, host water homeostasis and water channels aquaporins (AQP) are critical for cell morphology and functions as AQP interact indirectly with the cell cytoskeleton and signaling cascades. Here, we investigated how P. aeruginosa 3O-C12-HSL affects cell morphology, area, volume and AQP9 expression and distribution in human primary macrophages, using quantitative PCR, immunoblotting, two- and three-dimensional live imaging, confocal and nanoscale imaging. Thus, 3O-C12-HSL enhanced cell volume and area and induced cell shape and protrusion fluctuations in macrophages, processes tentatively driven by fluxes of water across cell membrane through AQP9, the predominant AQP in macrophages. Moreover, 3O-C12-HSL upregulated the expression of AQP9 at both the protein and mRNA levels. This was accompanied with enhanced whole cell AQP9 fluorescent intensity and redistribution of AQP9 to the leading and trailing regions, in parallel with increased cell area in the macrophages. Finally, nanoscopy imaging provided details on AQP9 dynamics and architecture within the lamellipodial area of 3O-C12-HSL-stimulated cells. We suggest that these novel events in the interaction between P. aeruginosa and macrophage may have an impact on the effectiveness of innate immune cells to fight bacteria, and thereby resolve the early stages of infections and inflammations.

  18. Pseudomonas aeruginosa N-3-oxo-dodecanoyl-homoserine Lactone Elicits Changes in Cell Volume, Morphology, and AQP9 Characteristics in Macrophages

    PubMed Central

    Holm, Angelika; Magnusson, Karl-Eric; Vikström, Elena

    2016-01-01

    Quorum sensing (QS) communication allows Pseudomonas aeruginosa to collectively control its population density and the production of biofilms and virulence factors. QS signal molecules, like N-3-oxo-dodecanoyl-L-homoserine lactone (3O-C12-HSL), can also affect the behavior of host cells, e.g., by modulating the chemotaxis, migration, and phagocytosis of human leukocytes. Moreover, host water homeostasis and water channels aquaporins (AQP) are critical for cell morphology and functions as AQP interact indirectly with the cell cytoskeleton and signaling cascades. Here, we investigated how P. aeruginosa 3O-C12-HSL affects cell morphology, area, volume and AQP9 expression and distribution in human primary macrophages, using quantitative PCR, immunoblotting, two- and three-dimensional live imaging, confocal and nanoscale imaging. Thus, 3O-C12-HSL enhanced cell volume and area and induced cell shape and protrusion fluctuations in macrophages, processes tentatively driven by fluxes of water across cell membrane through AQP9, the predominant AQP in macrophages. Moreover, 3O-C12-HSL upregulated the expression of AQP9 at both the protein and mRNA levels. This was accompanied with enhanced whole cell AQP9 fluorescent intensity and redistribution of AQP9 to the leading and trailing regions, in parallel with increased cell area in the macrophages. Finally, nanoscopy imaging provided details on AQP9 dynamics and architecture within the lamellipodial area of 3O-C12-HSL-stimulated cells. We suggest that these novel events in the interaction between P. aeruginosa and macrophage may have an impact on the effectiveness of innate immune cells to fight bacteria, and thereby resolve the early stages of infections and inflammations. PMID:27047801

  19. Metre-long cell-laden microfibres exhibit tissue morphologies and functions

    NASA Astrophysics Data System (ADS)

    Onoe, Hiroaki; Okitsu, Teru; Itou, Akane; Kato-Negishi, Midori; Gojo, Riho; Kiriya, Daisuke; Sato, Koji; Miura, Shigenori; Iwanaga, Shintaroh; Kuribayashi-Shigetomi, Kaori; Matsunaga, Yukiko T.; Shimoyama, Yuto; Takeuchi, Shoji

    2013-06-01

    Artificial reconstruction of fibre-shaped cellular constructs could greatly contribute to tissue assembly in vitro. Here we show that, by using a microfluidic device with double-coaxial laminar flow, metre-long core-shell hydrogel microfibres encapsulating ECM proteins and differentiated cells or somatic stem cells can be fabricated, and that the microfibres reconstitute intrinsic morphologies and functions of living tissues. We also show that these functional fibres can be assembled, by weaving and reeling, into macroscopic cellular structures with various spatial patterns. Moreover, fibres encapsulating primary pancreatic islet cells and transplanted through a microcatheter into the subrenal capsular space of diabetic mice normalized blood glucose concentrations for about two weeks. These microfibres may find use as templates for the reconstruction of fibre-shaped functional tissues that mimic muscle fibres, blood vessels or nerve networks in vivo.

  20. Solvent polarity and nanoscale morphology in bulk heterojunction organic solar cells: A case study

    SciTech Connect

    Thomas, Ajith; Elsa Tom, Anju; Ison, V. V. E-mail: praveen@materials.iisc.ernet.in; Rao, Arun D.; Varman, K. Arul; Ranjith, K.; Ramamurthy, Praveen C. E-mail: praveen@materials.iisc.ernet.in; Vinayakan, R.

    2014-03-14

    Organic bulk heterojunction solar cells were fabricated under identical experimental conditions, except by varying the solvent polarity used for spin coating the active layer components and their performance was evaluated systematically. Results showed that presence of nitrobenzene-chlorobenzene composition governs the morphology of active layer formed, which is due to the tuning of solvent polarity as well as the resulting solubility of the P3HT:PCBM blend. Trace amount of nitrobenzene favoured the formation of better organised P3HT domains, as evident from conductive AFM, tapping mode AFM and surface, and cross-sectional SEM analysis. The higher interfacial surface area thus generated produced cells with high efficiency. But, an increase in the nitrobenzene composition leads to a decrease in cell performance, which is due to the formation of an active layer with larger size polymer domain networks with poor charge separation possibility.

  1. Using wavelet denoising and mathematical morphology in the segmentation technique applied to blood cells images.

    PubMed

    Boix, Macarena; Cantó, Begoña

    2013-04-01

    Accurate image segmentation is used in medical diagnosis since this technique is a noninvasive pre-processing step for biomedical treatment. In this work we present an efficient segmentation method for medical image analysis. In particular, with this method blood cells can be segmented. For that, we combine the wavelet transform with morphological operations. Moreover, the wavelet thresholding technique is used to eliminate the noise and prepare the image for suitable segmentation. In wavelet denoising we determine the best wavelet that shows a segmentation with the largest area in the cell. We study different wavelet families and we conclude that the wavelet db1 is the best and it can serve for posterior works on blood pathologies. The proposed method generates goods results when it is applied on several images. Finally, the proposed algorithm made in MatLab environment is verified for a selected blood cells.

  2. Genetic background affects susceptibility to tumoral stem cell reprogramming

    PubMed Central

    García-Ramírez, Idoia; Ruiz-Roca, Lucía; Martín-Lorenzo, Alberto; Blanco, Óscar; García-Cenador, María Begoña; García-Criado, Francisco Javier; Vicente-Dueñas, Carolina; Sánchez-García, Isidro

    2013-01-01

    The latest studies of the interactions between oncogenes and its target cell have shown that certain oncogenes may act as passengers to reprogram tissue-specific stem/progenitor cell into a malignant cancer stem cell state. In this study, we show that the genetic background influences this tumoral stem cell reprogramming capacity of the oncogenes using as a model the Sca1-BCRABLp210 mice, where the type of tumor they develop, chronic myeloid leukemia (CML), is a function of tumoral stem cell reprogramming. Sca1-BCRABLp210 mice containing FVB genetic components were significantly more resistant to CML. However, pure Sca1-BCRABLp210 FVB mice developed thymomas that were not seen in the Sca1-BCRABLp210 mice into the B6 background. Collectively, our results demonstrate for the first time that tumoral stem cell reprogramming fate is subject to polymorphic genetic control. PMID:23839033

  3. Harvesting Technique Affects Adipose-Derived Stem Cell Yield

    PubMed Central

    Iyyanki, Tejaswi; Hubenak, Justin; Liu, Jun; Chang, Edward I.; Beahm, Elisabeth K.; Zhang, Qixu

    2015-01-01

    Background The success of an autologous fat graft depends in part on its total stromal vascular fraction (SVF) and adipose-derived stem cells (ASCs). However, variations in the yields of ASCs and SVF cells as a result of different harvesting techniques and donor sites are poorly understood. Objective To investigate the effects of adipose tissue harvesting technique and donor site on the yield of ASCs and SVF cells. Methods Subcutaneous fat tissues from the abdomen, flank, or axilla were harvested from patients of various ages by mechanical liposuction, direct surgical excision, or Coleman's technique with or without centrifugation. Cells were isolated and then analyzed with flow cytometry to determine the yields of total SVF cells and ASCs (CD11b−, CD45−, CD34+, CD90+, D7-FIB+). Differences in ASC and total SVF yields were assessed with one-way analysis of variance. Differentiation experiments were performed to confirm the multilineage potential of cultured SVF cells. Results Compared with Coleman's technique without centrifugation, direct excision yielded significantly more ASCs (P < .001) and total SVF cells (P = .007); liposuction yielded significantly fewer ASCs (P < .001) and total SVF cells (P < .05); and Coleman's technique with centrifugation yielded significantly more total SVF cells (P < .005), but not ASCs. The total number of SVF cells in fat harvested from the abdomen was significantly larger than the number in fat harvested from the flank or axilla (P < .05). Cultured SVF cells differentiated to adipocytes, osteocytes, and chondrocytes. Conclusions Adipose tissue harvested from the abdomen through direct excision or Coleman's technique with centrifugation was found to yield the most SVF cells and ASCs. PMID:25791999

  4. Topochemical and morphological characterization of wood cell wall treated with the ionic liquid, 1-ethylpyridinium bromide.

    PubMed

    Kanbayashi, Toru; Miyafuji, Hisashi

    2015-09-01

    MAIN CONCLUSION : [EtPy][Br] is more reactive toward lignin than toward the PSs in wood cell walls, and [EtPy][Br] treatment results in inhomogenous changes to the cell wall's ultrastructural and chemical components. The effects of the ionic liquid 1-ethylpyridinium bromide ([EtPy][Br]), which prefers to react with lignin rather than cellulose on the wood cell walls of Japanese cedar (Cryptomeria japonica), were investigated from a morphology and topochemistry point of view. The [EtPy][Br] treatment induced cell wall swelling, the elimination of warts, and the formation of countless pores in the tracheids. However, many of the pit membranes and the cellulose crystalline structure remained unchanged. Raman microscopic analyses revealed that chemical changes in the cell walls were different for different layers and that the lignin in the compound middle lamella and the cell corner resists interaction with [EtPy][Br]. Additionally, the interaction of [EtPy][Br] with the wood cell wall is different to that of other types of ionic liquid. PMID:25556160

  5. Developmental Profile, Morphology, and Synaptic Connectivity of Cajal-Retzius Cells in the Postnatal Mouse Hippocampus.

    PubMed

    Anstötz, Max; Huang, Hao; Marchionni, Ivan; Haumann, Iris; Maccaferri, Gianmaria; Lübke, Joachim H R

    2016-02-01

    Cajal-Retzius (CR) cells are early generated neurons, involved in the assembly of developing neocortical and hippocampal circuits. However, their roles in networks of the postnatal brain remain poorly understood. In order to get insights into these latter functions, we have studied their morphological and synaptic properties in the postnatal hippocampus of the CXCR4-EGFP mouse, where CR cells are easily identifiable. Our data indicate that CR cells are nonuniformly distributed along different subfields of the hippocampal formation, and that their postnatal decline is regulated in a region-specific manner. In fact, CR cells persist in distinct areas of fully mature animals. Subclasses of CR cells project and target either local (molecular layers) or distant regions [subicular complex and entorhinal cortex (EC)] of the hippocampal formation, but have similar firing patterns. Lastly, CR cells are biased toward targeting dendritic shafts compared with spines, and produce large-amplitude glutamatergic unitary postsynaptic potentials on γ-aminobutyric acid (GABA) containing interneurons. Taken together, our results suggest that CR cells are involved in a novel excitatory loop of the postnatal hippocampal formation, which potentially contributes to shaping the flow of information between the hippocampus, parahippocampal regions and entorhinal cortex, and to the low seizure threshold of these brain areas.

  6. Feeding Frequency Affects Cultured Rat Pituitary Cells in Low Gravity

    NASA Technical Reports Server (NTRS)

    Hymer, W. C.; Grindeland, R. E.; Salada, T.; Cenci, R.; Krishnan, K.; Mukai, C.; Nagaoka, S.

    1996-01-01

    In this report, we describe the results of a rat pituitary cell culture experiment done on STS-65 in which the effect of cell feeding on the release of the six anterior pituitary hormones was studied. We found complex microgravity related interactions between the frequency of cell feeding and the quantity and quality (i.e. biological activity) of some of the six hormones released in flight. Analyses of growth hormone (GH) released from cells into culture media on different mission days using gel filtration and ion exchange chromatography yielded qualitatively similar results between ground and flight samples. Lack of cell feeding resulted in extensive cell clumping in flight (but not ground) cultures. Vigorous fibroblast growth occurred in both ground and flight cultures fed 4 times. These results are interpreted within the context of autocrine and or paracrine feedback interactions. Finally the payload specialist successfully prepared a fresh trypsin solution in microgravity, detached the cells from their surface and reinserted them back into the culture chamber. These cells reattached and continued to release hormone in microgravity. In summary, this experiment shows that pituitary cells are microgravity sensitive and that coupled operations routinely associated with laboratory cel1 culture can also be accomplished in low gravity.

  7. From Morphology to Interfaces to Tandem Geometries: Enhancing the Performance of Perovskite/Polymer Solar Cells

    NASA Astrophysics Data System (ADS)

    Russell, Thomas

    We have taken a new approach to develop mesoporous lead iodide scaffolds, using the nucleation and growth of lead iodide crystallites in a wet film. A simple time-dependent growth control enabled the manipulation of the mesoporous lead iodide layer quality in a continuous manner. The morphology of lead iodide is shown to influence the subsequent crystallization of methyamoniumleadiodide film by using angle-dependent grazing incidence x-ray scattering. The morphology of lead iodide film can be fine-tuned, and thus the methyamoniumleadiodide film quality can be effectively controlled, leading to an optimization of the perovskite active layer. Using this strategy, perovskite solar cells with inverted PHJ structure showed a PCE of 15.7 per cent with little hysteresis. Interface engineering is critical for achieving efficient solar cells, yet a comprehensive understanding of the interface between metal electrode and electron transport layer (ETL) is lacking. A significant power conversion efficiency (PCE) improvement of fullerene/perovskite planar heterojunction solar cells was achieved by inserting a fulleropyrrolidine interlayer between the silver electrode and electron transport layer. The interlayer was found to enhance recombination resistance, increases electron extraction rate and prolongs free carrier lifetime. We also uncovered a facile solution-based fabrication of high performance tandem perovskite/polymer solar cells where the front sub-cell consists of perovskite and the back sub-cell is a polymer-based layer. A record maximum PCE of 15.96 per cent was achieved, demonstrating the synergy between the perovskite and semiconducting polymers. This design balances the absorption of the perovskite and the polymer, eliminates the adverse impact of thermal annealing during perovskite fabrication, and affords devices with no hysteresis. This work was performed in collaboration with Y. Liu, Z. Page, D. Venkataraman and T. Emrick (UMASS), F. Liu (LBNL) and Q. Hu and R

  8. Morphologic Damage of Rat Alveolar Epithelial Type II Cells Induced by Bile Acids Could Be Ameliorated by Farnesoid X Receptor Inhibitor Z-Guggulsterone In Vitro

    PubMed Central

    Huang, Yaowei; Hou, Xusheng; Wu, Wenyu; Nie, Lei; Tian, Yinghong; Lu, Yanmeng

    2016-01-01

    Objective. To determine whether bile acids (BAs) affect respiratory functions through the farnesoid X receptor (FXR) expressed in the lungs and to explore the possible mechanisms of BAs-induced respiratory disorder. Methods. Primary cultured alveolar epithelial type II cells (AECIIs) of rat were treated with different concentrations of chenodeoxycholic acid (CDCA) in the presence or absence of FXR inhibitor Z-guggulsterone (GS). Then, expression of FXR in nuclei of AECIIs was assessed by immunofluorescence microscopy. And ultrastructural changes of the cells were observed under transmission electron microscope and analyzed by Image-Pro Plus software. Results. Morphologic damage of AECIIs was exhibited in high BAs group in vitro, with high-level expression of FXR, while FXR inhibitor GS could attenuate the cytotoxicity of BAs to AECIIs. Conclusions. FXR expression was related to the morphologic damage of AECIIs induced by BAs, thus influencing respiratory functions. PMID:27340672

  9. Brucella abortus Choloylglycine Hydrolase Affects Cell Envelope Composition and Host Cell Internalization

    PubMed Central

    Marchesini, María Inés; Connolly, Joseph; Delpino, María Victoria; Baldi, Pablo C.; Mujer, Cesar V.; DelVecchio, Vito G.; Comerci, Diego J.

    2011-01-01

    Choloylglycine hydrolase (CGH, E.C. 3.5.1.24) is a conjugated bile salt hydrolase that catalyses the hydrolysis of the amide bond in conjugated bile acids. Bile salt hydrolases are expressed by gastrointestinal bacteria, and they presumably decrease the toxicity of host's conjugated bile salts. Brucella species are the causative agents of brucellosis, a disease affecting livestock and humans. CGH confers Brucella the ability to deconjugate and resist the antimicrobial action of bile salts, contributing to the establishment of a successful infection through the oral route in mice. Additionally, cgh-deletion mutant was also attenuated in intraperitoneally inoculated mice, which suggests that CGH may play a role during systemic infection other than hydrolyzing conjugated bile acids. To understand the role CGH plays in B. abortus virulence, we infected phagocytic and epithelial cells with a cgh-deletion mutant (Δcgh) and found that it is defective in the internalization process. This defect along with the increased resistance of Δcgh to the antimicrobial action of polymyxin B, prompted an analysis of the cell envelope of this mutant. Two-dimensional electrophoretic profiles of Δcgh cell envelope-associated proteins showed an altered expression of Omp2b and different members of the Omp25/31 family. These results were confirmed by Western blot analysis with monoclonal antibodies. Altogether, the results indicate that Brucella CGH not only participates in deconjugation of bile salts but also affects overall membrane composition and host cell internalization. PMID:22174816

  10. Brucella abortus choloylglycine hydrolase affects cell envelope composition and host cell internalization.

    PubMed

    Marchesini, María Inés; Connolly, Joseph; Delpino, María Victoria; Baldi, Pablo C; Mujer, Cesar V; DelVecchio, Vito G; Comerci, Diego J

    2011-01-01

    Choloylglycine hydrolase (CGH, E.C. 3.5.1.24) is a conjugated bile salt hydrolase that catalyses the hydrolysis of the amide bond in conjugated bile acids. Bile salt hydrolases are expressed by gastrointestinal bacteria, and they presumably decrease the toxicity of host's conjugated bile salts. Brucella species are the causative agents of brucellosis, a disease affecting livestock and humans. CGH confers Brucella the ability to deconjugate and resist the antimicrobial action of bile salts, contributing to the establishment of a successful infection through the oral route in mice. Additionally, cgh-deletion mutant was also attenuated in intraperitoneally inoculated mice, which suggests that CGH may play a role during systemic infection other than hydrolyzing conjugated bile acids. To understand the role CGH plays in B. abortus virulence, we infected phagocytic and epithelial cells with a cgh-deletion mutant (Δcgh) and found that it is defective in the internalization process. This defect along with the increased resistance of Δcgh to the antimicrobial action of polymyxin B, prompted an analysis of the cell envelope of this mutant. Two-dimensional electrophoretic profiles of Δcgh cell envelope-associated proteins showed an altered expression of Omp2b and different members of the Omp25/31 family. These results were confirmed by Western blot analysis with monoclonal antibodies. Altogether, the results indicate that Brucella CGH not only participates in deconjugation of bile salts but also affects overall membrane composition and host cell internalization.

  11. Morphology and ultrastructure of Interfilum and Klebsormidium (Klebsormidiales, Streptophyta) with special reference to cell division and thallus formation

    PubMed Central

    Mikhailyuk, Tatiana; Holzinger, Andreas; Massalski, Andrzej; Karsten, Ulf

    2014-01-01

    Representatives of the closely related genera, Interfilum and Klebsormidium, are characterized by unicells, dyads or packets in Interfilum and contrasting uniseriate filaments in Klebsormidium. According to the literature, these distinct thallus forms originate by different types of cell division, sporulation (cytogony) versus vegetative cell division (cytotomy), but investigations of their morphology and ultrastructure show a high degree of similarity. Cell walls of both genera are characterized by triangular spaces between cell walls of neighbouring cells and the parental wall or central space among the walls of a cell packet, exfoliations and projections of the parental wall and cap-like and H-like fragments of the cell wall. In both genera, each cell has its individual cell wall and it also has part of the common parental wall or its remnants. Therefore, vegetative cells of Interfilum and Klebsormidium probably divide by the same type of cell division (sporulation-like). Various strains representing different species of the two genera are characterized by differences in cell wall ultrastructure, particularly the level of preservation, rupture or gelatinization of the parental wall surrounding the daughter cells. The differing morphologies of representatives of various lineages result from features of the parental wall during cell separation and detachment. Cell division in three planes (usual in Interfilum and a rare event in Klebsormidium) takes place in spherical or short cylindrical cells, with the chloroplast positioned perpendicularly or obliquely to the filament (dyad) axis. The morphological differences are mainly a consequence of differing fates of the parental wall after cell division and detachment. The development of different morphologies within the two genera mostly depends on characters such as the shape of cells, texture of cell walls, mechanical interactions between cells and the influence of environmental conditions. PMID:26504365

  12. Sperm Associated Antigen 6 (SPAG6) Regulates Fibroblast Cell Growth, Morphology, Migration and Ciliogenesis.

    PubMed

    Li, Wei; Mukherjee, Abir; Wu, Jinhua; Zhang, Ling; Teves, Maria E; Li, Hongfei; Nambiar, Shanti; Henderson, Scott C; Horwitz, Alan R; Strauss Iii, Jerome F; Fang, Xianjun; Zhang, Zhibing

    2015-01-01

    Mammalian Spag6 is the orthologue of Chlamydomonas PF16, which encodes a protein localized in the axoneme central apparatus, and regulates flagella/cilia motility. Most Spag6-deficient mice are smaller in size than their littermates. Because SPAG6 decorates microtubules, we hypothesized that SPAG6 has other roles related to microtubule function besides regulating flagellar/cilia motility. Mouse embryonic fibroblasts (MEFs) were isolated from Spag6-deficient and wild-type embryos for these studies. Both primary and immortalized Spag6-deficient MEFs proliferated at a much slower rate than the wild-type MEFs, and they had a larger surface area. Re-expression of SPAG6 in the Spag6-deficient MEFs rescued the abnormal cell morphology. Spag6-deficient MEFs were less motile than wild-type MEFs, as shown by both chemotactic analysis and wound-healing assays. Spag6-deficient MEFs also showed reduced adhesion associated with a non-polarized F-actin distribution. Multiple centrosomes were observed in the Spag6-deficient MEF cultures. The percentage of cells with primary cilia was significantly reduced compared to the wild-type MEFs, and some Spag6-deficient MEFs developed multiple cilia. Furthermore, SPAG6 selectively increased expression of acetylated tubulin, a microtubule stability marker. The Spag6-deficient MEFs were more sensitive to paclitaxel, a microtubule stabilizer. Our studies reveal new roles for SPAG6 in modulation of cell morphology, proliferation, migration, and ciliogenesis. PMID:26585507

  13. Automated counting of morphologically normal red blood cells by using digital holographic microscopy and statistical methods

    NASA Astrophysics Data System (ADS)

    Moon, Inkyu; Yi, Faliu

    2015-09-01

    In this paper we overview a method to automatically count morphologically normal red blood cells (RBCs) by using off-axis digital holographic microscopy and statistical methods. Three kinds of RBC are used as training and testing data. All of the RBC phase images are obtained with digital holographic microscopy (DHM) that is robust to transparent or semitransparent biological cells. For the determination of morphologically normal RBCs, the RBC's phase images are first segmented with marker-controlled watershed transform algorithm. Multiple features are extracted from the segmented cells. Moreover, the statistical method of Hotelling's T-square test is conducted to show that the 3D features from 3D imaging method can improve the discrimination performance for counting of normal shapes of RBCs. Finally, the classifier is designed by using statistical Bayesian algorithm and the misclassification rates are measured with leave-one-out technique. Experimental results show the feasibility of the classification method for calculating the percentage of each typical normal RBC shape.

  14. Morphological and histological characteristics of mammary dysplasias occurring in cell dissociation-derived murine mammary outgrowths

    SciTech Connect

    Ethier, S.P.; Adams, L.M.; Ullrich, R.L.

    1984-10-01

    The morphological and histological characteristics of ductal dysplasias that were observed in mammary outgrowths derived from monodispersed mammary cells of carcinogen-treated mice are described. Mammary outgrowths were derived by injecting either 10(4) or 10(5) enzymatically dissociated mammary cells, obtained from control or carcinogen-treated BALB/c mice, into gland-free mammary fat pads of syngeneic hosts. The mammary dysplasias observed varied considerably in morphological and histological characteristics. The majority of the lesions were ductal in origin and were associated with epithelial hyperplasia which ranged from mild hyperplasia, in which only a few extra layers of epithelium were present, to severe hyperplasia, in which the ducts and end buds were occluded and distended with epithelial cells. In addition, papillary and lobular lesions were observed which were also associated with varying degrees of hyperplasia. The range of mammary dysplasias observed in these outgrowths closely resembles that of lesions associated with the pathogenesis of mammary carcinoma in mice, rats, and humans.

  15. Sperm Associated Antigen 6 (SPAG6) Regulates Fibroblast Cell Growth, Morphology, Migration and Ciliogenesis

    PubMed Central

    Li, Wei; Mukherjee, Abir; Wu, Jinhua; Zhang, Ling; Teves, Maria E.; Li, Hongfei; Nambiar, Shanti; Henderson, Scott C.; Horwitz, Alan R.; Strauss III, Jerome F.; Fang, Xianjun; Zhang, Zhibing

    2015-01-01

    Mammalian Spag6 is the orthologue of Chlamydomonas PF16, which encodes a protein localized in the axoneme central apparatus, and regulates flagella/cilia motility. Most Spag6-deficient mice are smaller in size than their littermates. Because SPAG6 decorates microtubules, we hypothesized that SPAG6 has other roles related to microtubule function besides regulating flagellar/cilia motility. Mouse embryonic fibroblasts (MEFs) were isolated from Spag6-deficient and wild-type embryos for these studies. Both primary and immortalized Spag6-deficient MEFs proliferated at a much slower rate than the wild-type MEFs, and they had a larger surface area. Re-expression of SPAG6 in the Spag6-deficient MEFs rescued the abnormal cell morphology. Spag6-deficient MEFs were less motile than wild-type MEFs, as shown by both chemotactic analysis and wound-healing assays. Spag6-deficient MEFs also showed reduced adhesion associated with a non-polarized F-actin distribution. Multiple centrosomes were observed in the Spag6-deficient MEF cultures. The percentage of cells with primary cilia was significantly reduced compared to the wild-type MEFs, and some Spag6-deficient MEFs developed multiple cilia. Furthermore, SPAG6 selectively increased expression of acetylated tubulin, a microtubule stability marker. The Spag6-deficient MEFs were more sensitive to paclitaxel, a microtubule stabilizer. Our studies reveal new roles for SPAG6 in modulation of cell morphology, proliferation, migration, and ciliogenesis. PMID:26585507

  16. A rapid antimicrobial susceptibility test based on single-cell morphological analysis.

    PubMed

    Choi, Jungil; Yoo, Jungheon; Lee, Mincheol; Kim, Eun-Geun; Lee, Ji Soo; Lee, Seungok; Joo, Seik; Song, Sang Hoon; Kim, Eui-Chong; Lee, Jung Chan; Kim, Hee Chan; Jung, Yong-Gyun; Kwon, Sunghoon

    2014-12-17

    A rapid antibiotic susceptibility test (AST) is desperately needed in clinical settings for fast and appropriate antibiotic administration. Traditional ASTs, which rely on cell culture, are not suitable for urgent cases of bacterial infection and antibiotic resistance owing to their relatively long test times. We describe a novel AST called single-cell morphological analysis (SCMA) that can determine antimicrobial susceptibility by automatically analyzing and categorizing morphological changes in single bacterial cells under various antimicrobial conditions. The SCMA was tested with four Clinical and Laboratory Standards Institute standard bacterial strains and 189 clinical samples, including extended-spectrum β-lactamase-positive Escherichia coli and Klebsiella pneumoniae, imipenem-resistant Pseudomonas aeruginosa, methicillin-resistant Staphylococcus aureus, and vancomycin-resistant Enterococci from hospitals. The results were compared with the gold standard broth microdilution test. The SCMA results were obtained in less than 4 hours, with 91.5% categorical agreement and 6.51% minor, 2.56% major, and 1.49% very major discrepancies. Thus, SCMA provides rapid and accurate antimicrobial susceptibility data that satisfy the recommended performance of the U.S. Food and Drug Administration. PMID:25520395

  17. Transcriptional modulator ZBED6 affects cell cycle and growth of human colorectal cancer cells.

    PubMed

    Akhtar Ali, Muhammad; Younis, Shady; Wallerman, Ola; Gupta, Rajesh; Andersson, Leif; Sjöblom, Tobias

    2015-06-23

    The transcription factor ZBED6 (zinc finger, BED-type containing 6) is a repressor of IGF2 whose action impacts development, cell proliferation, and growth in placental mammals. In human colorectal cancers, IGF2 overexpression is mutually exclusive with somatic mutations in PI3K signaling components, providing genetic evidence for a role in the PI3K pathway. To understand the role of ZBED6 in tumorigenesis, we engineered and validated somatic cell ZBED6 knock-outs in the human colorectal cancer cell lines RKO and HCT116. Ablation of ZBED6 affected the cell cycle and led to increased growth rate in RKO cells but reduced growth in HCT116 cells. This striking difference was reflected in the transcriptome analyses, which revealed enrichment of cell-cycle-related processes among differentially expressed genes in both cell lines, but the direction of change often differed between the cell lines. ChIP sequencing analyses displayed enrichment of ZBED6 binding at genes up-regulated in ZBED6-knockout clones, consistent with the view that ZBED6 modulates gene expression primarily by repressing transcription. Ten differentially expressed genes were identified as putative direct gene targets, and their down-regulation by ZBED6 was validated experimentally. Eight of these genes were linked to the Wnt, Hippo, TGF-β, EGF receptor, or PI3K pathways, all involved in colorectal cancer development. The results of this study show that the effect of ZBED6 on tumor development depends on the genetic background and the transcriptional state of its target genes.

  18. Phenotypic, Morphological and Adhesive Differences of Human Hematopoietic Progenitor Cells Cultured on Murine versus Human Mesenchymal Stromal Cells

    PubMed Central

    Reichert, Doreen; Friedrichs, Jens; Ritter, Steffi; Käubler, Theresa; Werner, Carsten; Bornhäuser, Martin; Corbeil, Denis

    2015-01-01

    Xenogenic transplantation models have been developed to study human hematopoiesis in immunocompromised murine recipients. They still have limitations and therefore it is important to delineate all players within the bone marrow that could account for species-specific differences. Here, we evaluated the proliferative capacity, morphological and physical characteristics of human CD34+ hematopoietic stem and progenitor cells (HSPCs) after co-culture on murine or human bone marrow-derived mesenchymal stromal cells (MSCs). After seven days, human CD34+CD133– HSPCs expanded to similar extents on both feeder layers while cellular subsets comprising primitive CD34+CD133+ and CD133+CD34– phenotypes are reduced fivefold on murine MSCs. The number of migrating HSPCs was also reduced on murine cells suggesting that MSC adhesion influences cellular polarization of HSPC. We used atomic force microscopy-based single-cell force spectroscopy to quantify their adhesive interactions. We found threefold higher detachment forces of human HSPCs from murine MSCs compared to human ones. This difference is related to the N-cadherin expression level on murine MSCs since its knockdown abolished their differential adhesion properties with human HSPCs. Our observations highlight phenotypic, morphological and adhesive differences of human HSPCs when cultured on murine or human MSCs, which raise some caution in data interpretation when xenogenic transplantation models are used. PMID:26498381

  19. Preparation of nano-hydroxyapatite particles with different morphology and their response to highly malignant melanoma cells in vitro

    NASA Astrophysics Data System (ADS)

    Li, Bo; Guo, Bo; Fan, Hongsong; Zhang, Xingdong

    2008-11-01

    To investigate the effects of nano-hydroxyapatite (HA) particles with different morphology on highly malignant melanoma cells, three kinds of HA particles with different morphology were synthesized and co-cultured with highly malignant melanoma cells using phosphate-buffered saline (PBS) as control. A precipitation method with or without citric acid addition as surfactant was used to produce rod-like hydroxyapatite (HA) particles with nano- and micron size, respectively, and a novel oil-in-water emulsion method was employed to prepare ellipse-like nano-HA particles. Particle morphology and size distribution of the as prepared HA powders were characterized by transmission electron microscope (TEM) and dynamic light scattering technique. The nano- and micron HA particles with different morphology were co-cultured with highly malignant melanoma cells. Immunofluorescence analysis and MTT assay were employed to evaluate morphological change of nucleolus and proliferation of tumour cells, respectively. To compare the effects of HA particles on cell response, the PBS without HA particles was used as control. The experiment results indicated that particle nanoscale effect rather than particle morphology of HA was more effective for the inhibition on highly malignant melanoma cells proliferation.

  20. Mutations in Coliphage P1 Affecting Host Cell Lysis

    PubMed Central

    Walker, Jean Tweedy; Walker, Donald H.

    1980-01-01

    A total of 103 amber mutants of coliphage P1 were tested for lysis of nonpermissive cells. Of these, 83 caused cell lysis at the normal lysis time and have defects in particle morphogenesis. Five amber mutants, with mutations in the same gene (gene 2), caused premature lysis and may have a defect in a lysis regulator. Fifteen amber mutants were unable to cause cell lysis. Artificially lysed cells infected with five of these mutants produced viable phage particles, and phage particles were seen in thin sections of unlysed, infected cells. However, phage production by these mutants was not continued after the normal lysis time. We conclude that the defect of these five mutants is in a lysis function. The five mutations were found to be in the same gene (designated gene 17). The remaining 10 amber mutants, whose mutations were found to be in the same gene (gene 10), were also unable to cause cell lysis. They differed from those in gene 17 in that no viable phage particles were produced from artificially lysed cells, and no phage particles were seen in thin sections of unlysed, infected cells. We conclude that the gene 10 mutants cannot synthesize late proteins, and it is possible that gene 10 may code for a regulator of late gene expression for P1. Images PMID:16789200

  1. Glycosaminoglycans affect heparanase location in CHO cell lines.

    PubMed

    Piva, Maria B R; Suarez, Eloah R; Melo, Carina M; Cavalheiro, Renan P; Nader, Helena B; Pinhal, Maria A S

    2015-09-01

    Glycosaminoglycans (GAG) play a ubiquitous role in tissues and cells. In eukaryotic cells, heparan sulfate (HS) is initially degraded by an endo-β-glucuronidase called heparanase-1 (HPSE). HS oligosaccharides generated by the action of HPSE intensify the activity of signaling molecules, activating inflammatory response, tumor metastasis, and angiogenesis. The aim of the present study was to understand if sulfated GAG could modulate HPSE, since the mechanisms that regulate HPSE have not been completely defined. CHO-K1 cells were treated with 4-methylumbelliferone (4-MU) and sodium chlorate, to promote total inhibition of GAG synthesis, and reduce the sulfation pattern, respectively. The GAG profile of the wild CHO-K1 cells and CHO-745, deficient in xylosyltransferase, was determined after [(35)S]-sulfate labeling. HPSE expression was determined via real-time quantitative polymerase chain reaction. Total ablation of GAG with 4-MU in CHO-K1 inhibited HPSE expression, while the lack of sulfation had no effect. Interestingly, 4-MU had no effect in CHO-745 cells for these assays. In addition, a different enzyme location was observed in CHO-K1 wild-type cells, which presents HPSE mainly in the extracellular matrix, in comparison with the CHO-745 mutant cells, which is found in the cytoplasm. In view of our results, we can conclude that GAG are essential modulators of HPSE expression and location. Therefore, GAG profile could impact cell behavior mediated by the regulation of HPSE.

  2. Cell culture density affects the proliferation activity of human adipose tissue stem cells.

    PubMed

    Kim, Dae Seong; Lee, Myoung Woo; Ko, Young Jong; Chun, Yong Hoon; Kim, Hyung Joon; Sung, Ki Woong; Koo, Hong Hoe; Yoo, Keon Hee

    2016-01-01

    In this study, we investigated the effect of cell density on the proliferation activity of human mesenchymal stem cells (MSCs) derived from adipose tissue (AT-MSCs) over time in culture. Passage #4 (P4) and #12 (P12) AT-MSCs from two donors were plated at a density of 200 (culture condition 1, CC1) or 5000 (culture condition 2, CC2) cells cm(-2) . After 7 days of incubation, P4 and P12 AT-MSCs cultured in CC1 were thin and spindle-shaped, whereas those cultured in CC2 had extensive cell-to-cell contacts and an expanded cell volume. In addition, P4 and P12 AT-MSCs in CC1 divided more than three times, while those in CC2 divided less than once on average. Flow cytometric analysis using 5(6)-carboxyfluorescein diacetate N-succinimidyl ester dye showed that the fluorescence intensity of AT-MSCs was lower in CC1 than in CC2. Furthermore, expression of proliferation-associated genes, such as CDC45L, CDC20A and KIF20A, in P4 AT-MSCs was higher in CC1 than in CC2, and this difference was also observed in P12 AT-MSCs. These data demonstrated that cell culture density affects the proliferation activity of MSCs, suggesting that it is feasible to design a strategy to prepare suitable MSCs using specific culture conditions.

  3. Quantification of cell, actin, and nuclear DNA morphology with high-throughput microscopy and CalMorph.

    PubMed

    Okada, Hiroki; Ohnuki, Shinsuke; Ohya, Yoshikazu

    2015-04-01

    Automated image acquisition and processing systems have been developed to quantitatively describe yeast cell morphology. These systems are superior to the preceding qualitative methods in terms of reproducibility, as they completely avoid subjective recognition of images. Because high-throughput microscopy has enabled rapid production of numerous cellular images, reinforcement of high-performance and high-throughput automated image-processing techniques has been in increasing demand in the field of biology. This protocol describes how to use a high-throughput microscope in conjunction with the image-processing software CalMorph, which outputs more than 500 morphological parameters, for quantification of cell, actin, and nuclear DNA morphology.