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Sample records for cell volume determine

  1. Determination of cell electroporation in small-volume samples.

    Science.gov (United States)

    Saulis, Gintautas; Praneviciŭte, Rita

    2007-01-01

    Expose of cells to electric field pulses increases the cell membrane permeability. Intracellular potassium ions leak out of the cells through aqueous pores created in the membrane. This release is used here for the determination of the fraction of electroporated cells. To determine cell membrane electroporation in small-volume samples (40-50 miacrol), mini both potassium ion-selective and reference electrodes, with tip diameters of 1-1.5 mm and minimum immersion depths of 1 mm, were utilized. The obtained calibration graph was linear within the concentration range 0.2-100 mM. The slope was 50-51 and 53-56 mV per concentration order at 10-11 and 19-21 degrees C, respectively. Detection limit of the electrode was determined to be 0.05-0.08 mM, however, it was possible to work down to concentrations in the range of 0.01 mM. Experiments have been carried out on human erythrocytes exposed to a square-wave electric pulse with the duration of 0.1-2 ms. The extracellular potassium concentrations were in the range between 0.04-0.08 mM (intact cells) and 3-5 mM (100% electroporation). The obtained dependences of the fraction of electroporated cells on the pulse intensity were of a sigmoid shape. The dependence of the pulse amplitude required to electroporate 50% of cells on the pulse duration, obtained from the release of intracellular potassium ions, coincided with the one determined from the extent of hemolysis after 24 h-incubation at low temperature.

  2. Determination of micro-litre volumes with high accuracy for flow cytometric blood cell counting

    Science.gov (United States)

    Reitz, S.; Kummrow, A.; Kammel, M.; Neukammer, J.

    2010-07-01

    We have gravimetrically calibrated the volumes dispensed by 1 mL syringes in the range between 1 µL and 100 µL using ultra-pure water. Protocols are based on series of consecutive difference measurements of masses in order to precisely compensate for evaporation, being the most important disturbing quantity. We determined expanded uncertainties of volume measurements for glass syringes of typically 0.2% (expansion factor 2) when dispensing volumes of 10 µL. For polypropylene syringes, selected with respect to the manufacturer, expanded uncertainties of 0.25% (expansion factor 2) were observed. Calibrated syringes were applied for measuring concentrations of blood cells in a flow cytometer demonstrating the capability to determine reference measurement values. Since the direct interaction of blood cells and syringe walls may lead to cell adhesion, glass syringes as well as (disposable) polypropylene syringes were calibrated.

  3. Cell volume and geometric parameters determination in living cells using confocal microscopy and 3D reconstruction

    OpenAIRE

    sprotocols

    2015-01-01

    Authors: David Hevia, Aida Rodriguez-Garcia, Marta Alonso-Gervós, Isabel Quirós-González, Henar M Cimadevilla, Carmen Gómez-Cordovés, Rosa M Sainz & Juan C Mayo ### Abstract The protocol reported here describes a simple, easy, fast and reproducible method aimed to know the geometric parameters of living cells based on confocal laser scanning microscopy combined with 3D reconstruction software. Briefly, the method is based on intrinsic fluorescence properties of acridine orange (AO...

  4. 36 CFR 223.36 - Volume determination.

    Science.gov (United States)

    2010-07-01

    ... 36 Parks, Forests, and Public Property 2 2010-07-01 2010-07-01 false Volume determination. 223.36... Volume determination. (a) Timber sale contracts may provide for volume determination by scaling... the contract or permit provides for the determination of volume by tree measurement and the timber has...

  5. Does the intracellular ionic concentration or the cell water content (cell volume) determine the activity of TonEBP in NIH3T3 cells?

    DEFF Research Database (Denmark)

    Rødgaard, Tina; Schou, Kenneth; Friis, Martin Barfred

    2008-01-01

    of the present investigation was to investigate whether cell shrinkage or high intracellular ionic concentration induced the activation of TonEBP. We designed a model system for isotonically shrinking cells over a prolonged period of time. Cells swelled in hypotonic medium and performed a regulatory volume......Cl(-) co-transporter, and Gadolinium inhibited shrinkage-activated Na(+) channels. Cells remained shrunken for at least 4 hours (isotonically shrunken cells). The activity of TonEBP was investigated with a Luciferase assay after isotonic shrinkage and after shrinkage in a high NaCl hypertonic medium......(+) than isotonically shrunken cells. This strongly suggested that an increase in intracellular ionic concentration and not cell shrinkage is involved in TonEBP activation....

  6. Can body volume be determined by PET?

    Science.gov (United States)

    Hentschel, Michael; Paul, Dominik; Korsten-Reck, Ulrike; Mix, Michael; Müller, Frank; Merk, Stefan; Moser, Ernst; Brink, Ingo

    2005-05-01

    To avoid dependence on body weight, the standardised uptake value (SUV) in positron emission tomography (PET) can instead be normalised to the lean body mass (LBM), which can be determined from body volume and mass. This study was designed to answer the following questions: Firstly, can the total body volume in principle be determined using PET? Secondly, is the precision of this measurement comparable to that achieved using an established standard method. Ten patients were examined during oncological whole-body PET examinations. The whole-body volume of the patients was determined from the transmission scan in PET. Air displacement plethysmography with BOD POD was used for comparison as the standard method of volume determination. In all patients, the whole-body volumes could be determined using PET and the standard method. Bland and Altman [23] analysis for agreement between the volumes determined by the two methods (presentation of differences vs means) revealed a very small difference of -0.14 l. With a mean patient volume of 71.81+/-15.93 l, the relative systematic error is only <0.1%. On this basis, equality of the volume values determined by the two methods can be assumed. PET can be used as a supplementary method for experimental determination of whole-body volume and total body fat in tumour patients. The fat content can be used to calculate the LBM and to determine body weight-independent SUVs (SUV(LBM)).

  7. Can body volume be determined by PET?

    Energy Technology Data Exchange (ETDEWEB)

    Hentschel, Michael; Paul, Dominik; Mix, Michael; Moser, Ernst; Brink, Ingo [University Hospital Freiburg, Division of Nuclear Medicine, Section of Positron Emission Tomography, Freiburg (Germany); Korsten-Reck, Ulrike [University Hospital Freiburg, Division of Sports Medicine, Freiburg (Germany); Mueller, Frank [PET-Institute Rhein-Neckar, Ludwigshafen (Germany); Merk, Stefan [Kantonsspital Basel, Division of Nuclear Medicine, Basel (Switzerland)

    2005-04-01

    To avoid dependence on body weight, the standardised uptake value (SUV) in positron emission tomography (PET) can instead be normalised to the lean body mass (LBM), which can be determined from body volume and mass. This study was designed to answer the following questions: Firstly, can the total body volume in principle be determined using PET? Secondly, is the precision of this measurement comparable to that achieved using an established standard method. Ten patients were examined during oncological whole-body PET examinations. The whole-body volume of the patients was determined from the transmission scan in PET. Air displacement plethysmography with BOD POD was used for comparison as the standard method of volume determination. In all patients, the whole-body volumes could be determined using PET and the standard method. Bland and Altman [23] analysis for agreement between the volumes determined by the two methods (presentation of differences vs means) revealed a very small difference of -0.14 l. With a mean patient volume of 71.81{+-}15.93 l, the relative systematic error is only <0.1%. On this basis, equality of the volume values determined by the two methods can be assumed. PET can be used as a supplementary method for experimental determination of whole-body volume and total body fat in tumour patients. The fat content can be used to calculate the LBM and to determine body weight-independent SUVs (SUV{sub LBM}). (orig.)

  8. Ion Channels Involved in Cell Volume Regulation

    DEFF Research Database (Denmark)

    Hoffmann, Else Kay

    2011-01-01

    This mini review outlines studies of cell volume regulation in two closely related mammalian cell lines: nonadherent Ehrlich ascites tumour cells (EATC) and adherent Ehrlich Lettre ascites (ELA) cells. Focus is on the regulatory volume decrease (RVD) that occurs after cell swelling, the volume...

  9. Shear-induced Volume Decrease in MDCK Cells

    Science.gov (United States)

    Heo, Jinseok; Sachs, Frederick; Wang, Jianbin; Hua, Susan Z.

    2013-01-01

    Using a microfluidic cell volume sensor we measured the change in the cell volume of Madin-Darby Canine Kidney (MDCK) cells induced by shear stress. An increase in shear stress from 0.2 to 2.0 dyn/cm2 resulted in a volume decrease to a steady state volume ~ 20 – 30 % smaller than the initial resting cell volume. Independent experiments based on fluorescence quenching confirmed the volume reduction. This shear-induced cell shrinkage was irreversible on the time scale of the experiment (~ 30 min). Treatment of 0.1 μM Hg2+ significantly inhibited the volume decrease, suggesting that the shear-induced cell shrinkage is associated with water efflux through aquaporins. The volume decrease cannot be inhibited by 75 mM TEA, 100 μM DIDS, or 100 μM Gd3+ suggesting that volume reduction is not directly mediated by K+ and Cl− channels that typically function during regulatory volume decrease (RVD), nor is it through cationic stretch-activated ion channels (SACs). The process also appears to be Ca2+ independent because it was insensitive to intracellular Ca2+ level. Since cell volume is determined by the intracellular water content, we postulate that the shear induced reductions in cell volume may arise from increased intracellular hydrostatic pressure as the cell is deformed under flow, which promotes the efflux of water. The increase in internal pressure in a deformable object under the flow is supported by the finite element mechanical model. PMID:22759987

  10. Apparatus for determining changes in limb volume

    Science.gov (United States)

    Bhagat, P. K.; Wu, V. C. (Inventor)

    1981-01-01

    Measuring apparatus for determining changes in the volume of limbs or other boty extremities by determining the cross-sectional area of such limbs many comprise a transmitter including first and second transducers for positioning on the surface of the limb at a predetermined distance there between, and a receiver including a receiver crystal for positioning on the surface of the limb. The distance between the receiver crystal and the first and second transducers are represented by respective first and second chords of the cross-section of the limb and the predetermined distance between the first and second transducers is represented by a third chord of the limb cross section.

  11. Thermodynamics of Condensed Phases: Formula Unit Volume, "V[subscript m]", and the Determination of the Number of Formula Units, "Z", in a Crystallographic Unit Cell

    Science.gov (United States)

    Glasser, Leslie

    2011-01-01

    Formula unit (or molecular) volume, "V[subscript m]", is related to many thermodynamic and physical properties of materials, so that knowledge of "V[subscript m]" is useful in prediction of such properties for known and even hypothetical materials. The symbol "Z" represents the number of formula units in a crystallographic unit cell; "Z" thus…

  12. A revised and unified pressure-clamp/relaxation theory for studying plant cell water relations with pressure probes: in-situ determination of cell volume for calculation of volumetric elastic modulus and hydraulic conductivity.

    Science.gov (United States)

    Knipfer, T; Fei, J; Gambetta, G A; Shackel, K A; Matthews, M A

    2014-10-21

    The cell-pressure-probe is a unique tool to study plant water relations in-situ. Inaccuracy in the estimation of cell volume (νo) is the major source of error in the calculation of both cell volumetric elastic modulus (ε) and cell hydraulic conductivity (Lp). Estimates of νo and Lp can be obtained with the pressure-clamp (PC) and pressure-relaxation (PR) methods. In theory, both methods should result in comparable νo and Lp estimates, but this has not been the case. In this study, the existing νo-theories for PC and PR methods were reviewed and clarified. A revised νo-theory was developed that is equally valid for the PC and PR methods. The revised theory was used to determine νo for two extreme scenarios of solute mixing between the experimental cell and sap in the pressure probe microcapillary. Using a fully automated cell-pressure-probe (ACPP) on leaf epidermal cells of Tradescantia virginiana, the validity of the revised theory was tested with experimental data. Calculated νo values from both methods were in the range of optically determined νo (=1.1-5.0nL) for T. virginiana. However, the PC method produced a systematically lower (21%) calculated νo compared to the PR method. Effects of solute mixing could only explain a potential error in calculated νo of cell turgor) of 19%, which is a fundamental parameter in calculating νo. It followed from the revised theory that the ratio of ΔV/ΔP was inversely related to the solute reflection coefficient. This highlighted that treating the experimental cell as an ideal osmometer in both methods is potentially not correct. Effects of non-ideal osmotic behavior by transmembrane solute movement may be minimized in the PR as compared to the PC method.

  13. Physiology of cell volume regulation in vertebrates

    DEFF Research Database (Denmark)

    Hoffmann, Else K; Lambert, Ian H; Pedersen, Stine F

    2009-01-01

    cases, activation of volume regulatory osmolyte transport. After acute swelling, cell volume is regulated by the process of regulatory volume decrease (RVD), which involves the activation of KCl cotransport and of channels mediating K(+), Cl(-), and taurine efflux. Conversely, after acute shrinkage......, cell volume is regulated by the process of regulatory volume increase (RVI), which is mediated primarily by Na(+)/H(+) exchange, Na(+)-K(+)-2Cl(-) cotransport, and Na(+) channels. Here, we review in detail the current knowledge regarding the molecular identity of these transport pathways...... and their regulation by, e.g., membrane deformation, ionic strength, Ca(2+), protein kinases and phosphatases, cytoskeletal elements, GTP binding proteins, lipid mediators, and reactive oxygen species, upon changes in cell volume. We also discuss the nature of the upstream elements in volume sensing in vertebrate...

  14. Simultaneous determination of extracellular volume and blood volume with the Volemetron

    NARCIS (Netherlands)

    Planque, B.A. de; Geyskes, C.G.; Dongen, R. van; Dorhout Mees, E.J.

    A new instrument, the “Volemetron”***, constructed to measure blood volume with radioactive isotopes, was adapted to determine 82Br distribution volume. Details of the technique arc given. Mean values of both RISA and 82Br distribution volume in normal men and women were determined. They were in

  15. Mean corpuscular volume of control red blood cells determines the interpretation of eosin-5'-maleimide (EMA) test result in infants aged less than 6 months.

    Science.gov (United States)

    Ciepiela, Olga; Adamowicz-Salach, Anna; Bystrzycka, Weronika; Łukasik, Jan; Kotuła, Iwona

    2015-08-01

    Eosin-5'-maleimide (EMA) binding test is a flow cytometric test used to detect hereditary spherocytosis (HS). To perform the test sample from patients, 5-6 reference samples of red blood are needed. Our aim was to investigate how the mean corpuscular volume (MCV) of red blood cells influences on the value of fluorescence of bounded EMA dye and how the choice of reference samples affects the test result. EMA test was performed in peripheral blood from 404 individuals, including 31 children suffering from HS. Mean fluorescence channel of EMA-RBCs was measured with Cytomics FC500 flow cytometer. Mean corpuscular volume of RBCs was assessed with LH750 Beckman Coulter. Statistical analysis was performed using Graph Pad Prism. The correlation Spearman coefficient between mean channel of fluorescence of EMA-RBCs and MCV was r = 0.39, p result might be negative. Due to different MCV values of RBCs in infancy and ca. Three months later, EMA test in neonates might be interpreted falsely negative. Samples from children younger than 3 months old had EMA test result 86.1 ± 11.7 %, whereas same samples that analyzed 4.1 ± 2.1 later had results of 75.4 ± 4.5 %, p results; thus, we recommend selection of reference samples with MCV in range of ±2 fL compared to MCV of patient RBC's.

  16. Volume morphology of printable solar cells

    Directory of Open Access Journals (Sweden)

    Joachim Loos

    2010-10-01

    Full Text Available Printable polymer or hybrid solar cells (PSCs have the potential to become one of the leading technologies of the 21st century in conversion of sunlight to electrical energy. Because of their ease of processing from solution fast and low cost mass production of devices is possible in a roll-to-roll printing fashion. The performance of such printed devices, in turn, is determined to a large extent by the three-dimensional organization of the photoactive layer, i.e. layer where light is absorbed and converted into free electrical charges, and its contacts with the charge collecting electrodes. In this review I briefly introduce our current understanding of morphology-performance relationships in PSCs with specific focus on electron tomography as analytical tool providing volume information with nanometer resolution.

  17. Rotation Effects on the Target-Volume Margin Determination

    CERN Document Server

    Zhang, Qinghui; Chan, M; Song, Y; Burman, C

    2014-01-01

    Rotational setup errors are usually neglected in most clinical centers. An analytical formula is developed to determine the extra margin between clinical target volume (CTV) and planning target volume (PTV) to account for setup errors. The proposed formula corrects for both translational and rotational setup errors and then incorporated into margin determination for PTV.

  18. Combined left and right ventricular volume determination by radionuclide angiocardiography using double bolus and equilibrium technique

    DEFF Research Database (Denmark)

    Stokholm, K H; Stubgaard, M; Møgelvang, J;

    1990-01-01

    by indicator dilution. The radionuclide technique comprised four steps: (1) a first-pass study of right ventricle; (2) a bolus study of left ventricle; (3) an equilibrium study of left ventricle; (4) determination of the distribution volume of red blood cells. Absolute volumes of left ventricle were determined......Eighteen patients with ischaemic heart disease were studied. Left and right ventricular volumes including cardiac output (forward flow) were determined by radionuclide angiocardiography using a double bolus and equilibrium technique. As reference, cardiac output was simultaneously measured...... from steps 2 + 3 + 4. Absolute volumes of right ventricle were calculated from stroke volume and right ventricular ejection fraction (EF) which in turn was determined from step 1 by creating composite systolic and composite diastolic images. There was an acceptable agreement between stroke volume...

  19. Immunobiology of natural killer cells. Volume II

    Energy Technology Data Exchange (ETDEWEB)

    Lotzova, E.; Herberman, R.B.

    1986-01-01

    This book provides a review of natural killer (NK) cell-mediated immunity in humans and experimental animal system. Topics for the volume include: In vivo activities of NK cells against primary and metastatic tumors in experimental animals; involvement of NK cells in human malignant disease; impaired NK cell profile in leukemia patients; in vivo modulation of NK activity in cancer patients; implications of aberrant NK cell activity in nonmalignant, chronic diseases; NK cell role in regulation of the growth and functions of hemopoietic and lymphoid cells; NK cells active against viral, bacterial, protozoan, and fungal infections; cytokine secretion and noncytotoxic functions of human large granular lymphocytes; augmentation of NK activity; regulation of NK cell activity by suppressor cells; NK cell cloning technology and characteristics of NK cell clones; comparison of antibody-dependent cellular cytotoxicity (ADCC) and NK activity, and index.

  20. Cell swelling and volume regulation

    DEFF Research Database (Denmark)

    Hoffmann, Else Kay

    1992-01-01

    been reported. The role of the different channels was discussed. A taurine leak pathway is clearly activated after cell swelling both in astrocytes and in neurones. The relations between the effect of glutamate and cell swelling were discussed. Discussion on the clearance of potassium from...... the extracellular space was continued. Neither the spatial buffering mechanism nor the role of the Na+/K+ pump was discussed here, but additional possibilities for K+ removal were discussed. At a high extracellular K+ concentration there is a possibility of channel-mediated KCl uptake by the glia cells, and a Na......-K-2Cl cotransport system is present in astrocytes and seems to play a role at relatively low extracellular K+ concentrations (6-20 mM). The cotransport system is likely to be regulated, but little is known about its regulation in glia cells....

  1. Cell volume regulation: physiology and pathophysiology

    DEFF Research Database (Denmark)

    Lambert, I H; Hoffmann, E K; Pedersen, Stine Helene Falsig

    2008-01-01

    not only under physiological conditions, e.g. following accumulation of nutrients, during epithelial absorption/secretion processes, following hormonal/autocrine stimulation, and during induction of apoptosis, but also under pathophysiological conditions, e.g. hypoxia, ischaemia and hyponatremia....../hypernatremia. On the other hand, it has recently become clear that an increase or reduction in cell volume can also serve as a specific signal in the regulation of physiological processes such as transepithelial transport, cell migration, proliferation and death. Although the mechanisms by which cell volume perturbations...... are sensed are still far from clear, significant progress has been made with respect to the nature of the sensors, transducers and effectors that convert a change in cell volume into a physiological response. In the present review, we summarize recent major developments in the field, and emphasize...

  2. Sperm flagellum volume determines freezability in red deer spermatozoa

    National Research Council Canada - National Science Library

    Ros-Santaella, José Luis; Domínguez-Rebolledo, Alvaro Efrén; Garde, José Julián

    2014-01-01

    ... provides sperm motility. Here, for the first time, we determined the volumes of the flagellum structures in fresh epididymal red deer spermatozoa using a stereological method under phase contrast microscopy...

  3. Aquaporin expression and cell volume regulation in the SV40 immortalized rat submandibular acinar cell line.

    Science.gov (United States)

    Hansen, Ann-Kristin; Galtung, Hilde Kanli

    2007-03-01

    The amount of aquaporins present and the cellular ability to perform regulatory volume changes are likely to be important for fluid secretions from exocrine glands. In this work these phenomena were studied in an SV40 immortalized rat submandibular acinar cell line. The regulatory cell volume characteristics have not previously been determined in these cells. Cell volume regulation following hyposmotic exposure and aquaporin induction was examined with Coulter counter methodology, radioactive efflux studies, fura-2 fluorescence, and polymerase chain reaction and Western blot techniques. Cell volume regulation was inhibited by the K(+) channel antagonists quinine and BaCl(2) and the Cl(-) channel blocker 5-nitro-2-(3-phenypropylamino)benzoic acid. A concomitant increase in cellular (3)H-taurine release and Ca(2+) concentration was also observed. Chelation of both intra- and extracellular Ca(2+) with EGTA and the Ca(2+) ionophore A23187 did not, however, affect cell volume regulation. Aquaporin 5 (AQP5) mRNA and protein levels were upregulated in hyperosmotic conditions and downregulated upon return to isosmotic solutions, but were reduced by the mitogen-activated ERK-activating kinase (MEK) inhibitor U0126. A 24-h MEK inhibition also diminished hyposmotically induced cell swelling and cell volume regulation. In conclusion, it was determined that regulatory volume changes in this immortalized cell line are due to KCl and taurine efflux. In conditions that increased AQP5 levels, the cells showed a faster cell swelling and a more complete volume recovery following hyposmotic exposure. This response could be overturned by MEK inhibition.

  4. Determination of the autonomously functioning volume of the thyroid

    Energy Technology Data Exchange (ETDEWEB)

    Emrich, D.; Erlenmaier, U.; Pohl, M.; Luig, H. (Goettingen Univ. (Germany). Abt. fuer Nuklearmedizin)

    1993-05-01

    The aim of this work was to determine the autonomously functioning volume in euthyroid and hyperthyroid goitres for prognostic and therapeutic pruposes. To this end, various groups of patients were selected: Individuals without evidence of thyroid disease, euthyroid patients with diffuse goitre of normal structure and function, euthyroid patients with evidence of autonomy and patients with hyperthyroidism due to autonomy. In all of them the thyroid uptake of Technetium-99m was determined under exogeneous suppression (TcU[sub s]) in the euthyroid state and under endogenous suppression (TcU) in the hyperthyroid state. It was demonstrated that: 1. In patients with unifocal autonomy the TcU[sub s] and TcU correlated linearly with the autonomous volume delineated and measured by sonography. 2. A nearly identical result was obtained if the mean autonomous volume in individuals without thyroid disease of 2.2[+-]1.1 ml calculated by TcU[sub s]/TcU x total thyroid volume was used as a basis. 3. The critical autonomous volume, i.e. the volume at which hyperthryroidism will occur, was found to be 16 ml at a cumulated sensitivity and specificity of >0.9. The method can be used to select patients for definitive treatment before hyperthryroidism occurs and to measure the autonomously functioning volume independent of its distribution within the thyroid for treatment with radioiodine. The method is easy to perform and is also an example of how a relative parameter of a function can be converted into an absolute parameter of a functioning volume. (orig.).

  5. Measurement and determinants of infrarenal aortic thrombus volume

    Energy Technology Data Exchange (ETDEWEB)

    Golledge, Jonathan; Wolanski, Philippe [James Cook University, The Vascular Biology Unit, Townsville, Queensland (Australia); Townsville Hospital, Townsville, Queensland (Australia); Parr, Adam [James Cook University, The Vascular Biology Unit, Townsville, Queensland (Australia); Buttner, Petra [James Cook University, School of Public Health and Tropical Medicine, Townsville, Queensland (Australia)

    2008-09-15

    Intra-luminal thrombus has been suggested to play a role in the progression of abdominal aortic aneurysm (AAA). The aims of this study were twofold. Firstly, to assess the reproducibility of a computer tomography (CT)-based technique for measurement of aortic thrombus volume. Secondly, to examine the determinants of infrarenal aortic thrombus volume in a cohort of patients with aortic dilatation. A consecutive series of 75 patients assessed by CT angiography with maximum aortic diameter {>=}25 mm were recruited. Intra-luminal thrombus volume was measured by a semi-automated workstation protocol based on a previously defined technique to quantitate aortic calcification. Intra- and inter-observer reproducibility were assessed using correlation coefficients, coefficient of variation and Bland-Altman plots. Infrarenal aortic thrombus volume percentage was related to clinical, anatomical and blood characteristics of the patients using univariate and multivariate tests. Infrarenal aortic thrombus volume was related to the severity of aortic dilatation assessed by total aortic volume (r=0.87, P<0.0001) or maximum aortic diameter (r=0.74, P< 0.0001). We therefore examined the clinical determinates of aortic thrombus expressed as a percentage of total aortic volume. Aortic thrombus percentage was negatively correlated with serum high density lipoprotein (HDL, r=-0.31). By ordinal multiple logistic regression analysis serum HDL below median ({<=}1.2 mM) was associated with aortic thrombus percentage in the upper quartile adjusting for other risk factors (odds ratio 5.3, 95% CI 1.1-25.0). Infrarenal aortic thrombus volume can be measured reproducibly on CT. Serum HDL, which can be therapeutically raised, may play a role in discouraging aortic thrombus accumulation with implications in terms of delaying progression of AAA. (orig.)

  6. Determination of adrenal volume by MRI in healthy children

    DEFF Research Database (Denmark)

    Mouritsen, Annette; Johansen, Marie Lindhardt; Wohlfahrt-Veje, Christine

    2014-01-01

    BACKGROUND: Adrenal disorders such as congenital adrenal hyperplasia result in abnormal adrenal size and morphology, but little is known about the clinical value of magnetic resonance imaging (MRI) in determining adrenal volume. OBJECTIVE: To evaluate the potential usefulness of MR methodology...

  7. Determination of Thyroid Volume by Ultrasonography among Schoolchildren in Philippines

    Directory of Open Access Journals (Sweden)

    Bu Kyung Kim

    2012-01-01

    Full Text Available Objective. Iodine deficiency is defined by the goiter and the urinary iodine concentration. However, a lack of local thyroid volume reference data resulted in the vague definition of goiter, especially in school-aged children. The aim of this paper was to determine the thyroid volumes by ultrasonography in schoolchildren aged 6 to 12 years living in Cagayan areas in Philippine. Methods. Cross-sectional thyroid ultrasonographic data of 158 schoolchildren aged 6–12 years from Tuguegarao and Lagum in Cagayan valley, Philippine were used. Thyroid volumes were compared based on logistic issue and urban and rural area and compared with other previously reported data. Results. The mean values of thyroid volume in Tuguerago and Lagum were 2.99±1.34 mL and 2.42±0.92 mL. The thyroid size was significantly in association with age (P<0.00, weight (P<0.00, height (P<0.00, and BSA (P<0.00 by Pearson’s correlation. The median thyroid volumes of schoolchildren investigated in this study were generally low compared to international reference data by age group but not by BSA. Conclusions. We propose for the first time local reference ultrasound values for thyroid volumes in 6–12 aged schoolchildren that should be used for monitoring iodine deficiency disorders.

  8. Measurement of cell volume changes by fluorescence self-quenching

    DEFF Research Database (Denmark)

    Hamann, Steffen; Kiilgaard, J.F.; Litman, Thomas

    2002-01-01

    At high concentrations, certain fluorophores undergo self-quenching, i.e., fluorescence intensity decreases with increasing fluorophore concentration. Accordingly, the self-quenching properties can be used for measuring water volume changes in lipid vesicles. In cells, quantitative determination...... of water transport using fluorescence self-quenching has been complicated by the requirement of relatively high (mM) and often toxic loading concentrations. Here we report a simple method that uses low (muM) loading concentrations of calcein-acetoxymethyl ester (calcein-AM) to obtain intracellular...... concentrations of the fluorophore calcein suitable for measurement of changes in cell water volume by self-quenching. The relationship between calcein fluorescence intensity, when excited at 490 nm (its excitation maximum), and calcein concentration was investigated in vitro and in various cultured cell types...

  9. Impaired migration and cell volume regulation in aquaporin 5-deficient SPC-A1 cells.

    Science.gov (United States)

    Chen, Zhihong; Zhang, Ziqiang; Gu, Yutong; Bai, Chunxue

    2011-05-31

    Aquaporin 5 (AQP5) is widely expressed in various organ and tissues. In light of the novel oncogenic properties of aquaporins (AQPs), here we investigated the effect of AQP5 knockdown by RNAi on transmembrane osmotic water permeability, cell migration potential and cell volume regulation ability. AQP5 expression was inhibited by short hairpin RNA in SPC-A1 cells, a lung adenocarcinoma cell line. Cells loaded with a fluoroprobe (calcein-AM) were immersed in either isosmotic, hyperosmotic or hyposmotic solutions, and fluorescence intensity was recorded using confocal microscopy. These measurements were used to calculate osmotic water permeability coefficients (Pf) and to monitor regulated volume decrease (RVD). Tumor cell migration and invasion assays were performed in a modified Boyden chamber. Wound healing and colony forming ability were also tested. Although self-quenching was not found in SPC-A1 cells, we observed a linear relationship between fluorescence intensity and cell water volume, suggesting that this method is a sensitive and reproducible way to measure single-cell transmembrane water permeability. Cells in which the AQP5 gene was silenced showed a 49.4% decrease in osmotic water permeability, a 55.3% decrease in migration and a 28.4% decrease in invasion potential. In addition, RVD decreased remarkably with reduced osmotic water permeability. Our results suggest that AQP5, which mediates water permeability and thus regulates cell shape and volume, is a potentially important determinant in cell migration. Copyright © 2011 Elsevier B.V. All rights reserved.

  10. Automated three-dimensional single cell phenotyping of spindle dynamics, cell shape, and volume

    CERN Document Server

    Plumb, Kemp; Pelletier, Vincent; Kilfoil, Maria L

    2015-01-01

    We present feature finding and tracking algorithms in 3D in living cells, and demonstrate their utility to measure metrics important in cell biological processes. We developed a computational imaging hybrid approach that combines automated three-dimensional tracking of point-like features with surface determination from which cell (or nuclear) volume, shape, and planes of interest can be extracted. After validation, we applied the technique to real space context-rich dynamics of the mitotic spindle, and cell volume and its relationship to spindle length, in dividing living cells. These methods are additionally useful for automated segregation of pre-anaphase and anaphase spindle populations in budding yeast. We found that genetic deletion of the yeast kinesin-5 mitotic motor cin8 leads to large mother and daughter cells that were indistinguishable based on size, and that in those cells the spindle length becomes uncorrelated with cell size. The technique can be used to visualize and quantify tracked feature c...

  11. Determination of right ventricular volumes during aortic surgery.

    Science.gov (United States)

    Van der Linden, P; Gilbart, E; Engelman, E; de Rood, M; Vincent, J L

    1989-06-01

    The aim of the present study was to evaluate right ventricular (RV) preload by measurements of right ventricular volumes during aortic clamping and unclamping. Nine patients (aged 67 +/- 9 years) undergoing infrarenal aortic aneurysmectomy were monitored with a pulmonary artery catheter equipped with a fast-response thermistor, allowing determination of RV volumes by the thermodilution technique. Anesthesia consisted of a continuous infusion of alfentanil and 50% N2O. Aortic clamping resulted in a significant decrease in cardiac index (CI) and a significant increase in systemic vascular resistance (SVR). There was no significant change in right ventricular ejection fraction (RVEF) (from 35% +/- 6% to 33% +/- 8%) in the presence of a significant decrease in stroke index (from 37.2 +/- 9.8 to 31.1 +/- 10.0 mL/beat/m2, P less than 0.05), indicating a significant decrease in RV end-diastolic volume (from 106 +/- 17 to 92 +/- 19 mL, P less than 0.01). There were no significant changes in cardiac filling pressures. Aortic unclamping was associated with a significant increase in CI and a significant decrease in SVR. There were no significant changes in cardiac filling pressures, RVEF, or RV volumes. Measurements of RV volumes indicated that aortic clamping resulted in a decrease in RV preload, which is usually not demonstrated by measurements of right atrial pressure alone.

  12. Cell volume-regulated cation channels.

    Science.gov (United States)

    Wehner, Frank

    2006-01-01

    Considering the enormous turnover rates of ion channels when compared to carriers it is quite obvious that channel-mediated ion transport may serve as a rapid and efficient mechanism of cell volume regulation. Whenever studied in a quantitative fashion the hypertonic activation of non-selective cation channels is found to be the main mechanism of regulatory volume increase (RVI). Some channels are inhibited by amiloride (and may be related to the ENaC), others are blocked by Gd(3) and flufenamate (and possibly linked to the group of transient receptor potential (TRP) channels). Nevertheless, the actual architecture of hypertonicity-induced cation channels remains to be defined. In some preparations, hypertonic stress decreases K(+) channel activity so reducing the continuous K(+) leak out of the cell; this is equivalent to a net gain of cell osmolytes facilitating RVI. The hypotonic activation of K(+) selective channels appears to be one of the most common principles of regulatory volume decrease (RVD) and, in most instances, the actual channels involved could be identified on the molecular level. These are BKCa (or maxi K(+)) channels, IK(Ca) and SK(Ca) channels (of intermediate and small conductance, respectively), the group of voltage-gated (Kv) channels including their Beta (or Kv ancilliary) subunits, two-pore K(2P) channels, as well as inwardly rectifying K(+) (Kir) channels (also contributing to K(ATP) channels). In some cells, hypotonicity activates non-selective cation channels. This is surprising, at first sight, because of the inside negative membrane voltage and the sum of driving forces for Na(+) and K(+) diffusion across the cell membrane rather favouring net cation uptake. Some of these channels, however, exhibit a P(K)/P(Na) significantly higher than 1, whereas others are Ca(++) permeable linking hypotonic stress to the activation of Ca(++) dependent ion channels. In particular, the latter holds for the group of TRPs which are specialised in the

  13. Determination of volumes in pipeline drain operation; Determinacao de volumes em operacao de drenagem de oleodutos

    Energy Technology Data Exchange (ETDEWEB)

    Pires, Luis Fernando G.; Barreto, Claudio V.; Silva, Bruno G. de [Pontificia Univ. Catolica do Rio de Janeiro, RJ (Brazil). Dept. de Engenharia Mecanica. Nucleo de Simulacao Termohidraulica de Dutos - SIMDUT

    2005-07-01

    Usually, pipeline and equipment maintenance requires the partial or fully drainage of the line. To develop this operation, it is necessary to preview the volumes at the drainage points to allocate the available resources to remove the liquid. With this main objective it was developed a numerical tool that is able to calculate the drainage volumes using only the data source from the GIS profile. This is possible if we consider a pipeline at no-flow state where only the gravitational forces can act in the fluid. The present paper uses a spreadsheet to determine the product volumes based in the drain points sequence and considering no flow boundaries along the pipeline. To validate the methodology a comparison is done with the results acquired in two real state drainage operations. (author)

  14. Direct volume rendering methods for cell structures.

    Science.gov (United States)

    Martišek, Dalibor; Martišek, Karel

    2012-01-01

    The study of the complicated architecture of cell space structures is an important problem in biology and medical research. Optical cuts of cells produced by confocal microscopes enable two-dimensional (2D) and three-dimensional (3D) reconstructions of observed cells. This paper discuses new possibilities for direct volume rendering of these data. We often encounter 16 or more bit images in confocal microscopy of cells. Most of the information contained in these images is unsubstantial for the human vision. Therefore, it is necessary to use mathematical algorithms for visualization of such images. Present software tools as OpenGL or DirectX run quickly in graphic station with special graphic cards, run very unsatisfactory on PC without these cards and outputs are usually poor for real data. These tools are black boxes for a common user and make it impossible to correct and improve them. With the method proposed, more parameters of the environment can be set, making it possible to apply 3D filters to set the output image sharpness in relation to the noise. The quality of the output is incomparable to the earlier described methods and is worth increasing the computing time. We would like to offer mathematical methods of 3D scalar data visualization describing new algorithms that run on standard PCs very well.

  15. Blood volume measurements in gopher snakes, using autologous 51Cr-labeled red blood cells.

    Science.gov (United States)

    Smeller, J M; Bush, M; Seal, U S

    1978-02-01

    Blood volume determinations were performed in 5 anesthetized gopher snakes (Pituophis melanoleucus catenifer) by means of a 51Cr-labeled red blood cell (RBC) method. The mean blood volume was 52.8 ml/kg of body weight (+/- 6.21 SE). Previous blood volume measurements have not been reported for this species. The RBC survival rate was estimated to be greater than 660 days. The RBC survival rate is long, but it cannot be determined accurately by this method.

  16. Determinants of Left Atrial Volume in Patients with Atrial Fibrillation

    Science.gov (United States)

    Hochgruber, Thomas; Krisai, Philipp; Zimmermann, Andreas J.; Aeschbacher, Stefanie; Pumpol, Katrin; Kessel-Schaefer, Arnheid; Stephan, Frank-Peter; Handschin, Nadja; Sticherling, Christian; Osswald, Stefan; Kaufmann, Beat A.; Paré, Guillaume; Kühne, Michael; Conen, David

    2016-01-01

    Introduction Left atrial (LA) enlargement is an important risk factor for incident stroke and a key determinant for the success of rhythm control strategies in patients with atrial fibrillation (AF). However, factors associated with LA volume in AF patients remain poorly understood. Methods Patients with paroxysmal or persistent AF were enrolled in this study. Real time 3-D echocardiography was performed in all participants and analyzed offline in a standardized manner. We performed stepwise backward linear regression analyses using a broad set of clinical parameters to determine independent correlates for 3-D LA volume. Results We included 210 patients (70.9% male, mean age 61±11years). Paroxysmal and persistent AF were present in 95 (45%) and 115 (55%) patients, respectively. Overall, 115 (55%) had hypertension, 11 (5%) had diabetes, and 18 (9%) had ischemic heart disease. Mean indexed LA volume was 36±12ml/m2. In multivariable models, significant associations were found for female sex (β coefficient -10.51 (95% confidence interval (CI) -17.85;-3.16), p = 0.0053), undergoing cardioversion (β 11.95 (CI 5.15; 18.74), p = 0.0006), diabetes (β 14.23 (CI 2.36; 26.10), p = 0.019), body surface area (BSA) (β 34.21 (CI 19.30; 49.12), pglomerular filtration rate (β -0.21 (CI -0.36; -0.06), p = 0.0064) and plasma levels of NT-pro brain natriuretic peptide (NT-proBNP) (β 6.79 (CI 4.05; 9.52), p<0.0001), but not age (p = 0.59) or hypertension (p = 0.42). Our final model explained 52% of the LA volume variability. Conclusions In patients with AF, the most important correlates with LA volume are sex, BSA, diabetes, renal function and NT-proBNP, but not age or hypertension. These results may help to refine rhythm control strategies in AF patients. PMID:27701468

  17. Sperm flagellum volume determines freezability in red deer spermatozoa.

    Science.gov (United States)

    Ros-Santaella, José Luis; Domínguez-Rebolledo, Alvaro Efrén; Garde, José Julián

    2014-01-01

    The factors affecting the inter-individual differences in sperm freezability is a major line of research in spermatology. Poor sperm freezability is mainly characterised by a low sperm velocity, which in turn is associated with low fertility rates in most animal species. Studies concerning the implications of sperm morphometry on freezability are quite limited, and most of them are based on sperm head size regardless of the structural parts of the flagellum, which provides sperm motility. Here, for the first time, we determined the volumes of the flagellum structures in fresh epididymal red deer spermatozoa using a stereological method under phase contrast microscopy. Sperm samples from thirty-three stags were frozen and classified as good freezers (GF) or bad freezers (BF) at two hours post-thawing using three sperm kinetic parameters which are strongly correlated with fertility in this species. Fourteen stags were clearly identified as GF, whereas nineteen were BF. No significant difference in sperm head size between the two groups was found. On the contrary, the GF exhibited a lower principal piece volume than the BF (6.13 µm3 vs 6.61 µm3, respectively, p = 0.006). The volume of the flagellum structures showed a strong negative relationship with post-thawing sperm velocity. For instance, the volume of the sperm principal piece was negatively correlated with sperm velocity at two hours post-thawing (r = -0.60; psperm principal piece results in poor freezability, and highlights the key role of flagellum size in sperm cryopreservation success.

  18. Mechanisms underlying KCNQ1channel cell volume sensitivity

    DEFF Research Database (Denmark)

    Hammami, Sofia

    Cells are constantly exposed to changes in cell volume during cell metabolism, nutrient uptake, cell proliferation, cell migration and salt and water transport. In order to cope with these perturbations, potassium channels in line with chloride channels have been shown to be likely contributors...... to the process of cell volume adjustments. A great diversity of potassium channels being members of either the 6TM, 4 TM or 2 TM K+ channel gene family have been shown to be strictly regulated by small, fast changes in cell volume. However, the precise mechanism underlying the K+ channel sensitivity to cell...... mechanism of regulation. Besides being regulated by cell volume, KCNQ1 is also modulated by the interaction of the ß subunit KCNE1 giving rise to the cardiac IKs delayed rectifier potassium current. Apart from altering the kinetic characteristics of the KCNQ1 channel current, KCNE1 also augments the KCNQ1...

  19. Oil Formation Volume Factor Determination Through a Fused Intelligence

    Directory of Open Access Journals (Sweden)

    Gholami Amin

    2016-12-01

    Full Text Available Volume change of oil between reservoir condition and standard surface condition is called oil formation volume factor (FVF, which is very time, cost and labor intensive to determine. This study proposes an accurate, rapid and cost-effective approach for determining FVF from reservoir temperature, dissolved gas oil ratio, and specific gravity of both oil and dissolved gas. Firstly, structural risk minimization (SRM principle of support vector regression (SVR was employed to construct a robust model for estimating FVF from the aforementioned inputs. Subsequently, an alternating conditional expectation (ACE was used for approximating optimal transformations of input/output data to a higher correlated data and consequently developing a sophisticated model between transformed data. Eventually, a committee machine with SVR and ACE was constructed through the use of hybrid genetic algorithm-pattern search (GA-PS. Committee machine integrates ACE and SVR models in an optimal linear combination such that makes benefit of both methods. A group of 342 data points was used for model development and a group of 219 data points was used for blind testing the constructed model. Results indicated that the committee machine performed better than individual models.

  20. Oil Formation Volume Factor Determination Through a Fused Intelligence

    Science.gov (United States)

    Gholami, Amin

    2016-12-01

    Volume change of oil between reservoir condition and standard surface condition is called oil formation volume factor (FVF), which is very time, cost and labor intensive to determine. This study proposes an accurate, rapid and cost-effective approach for determining FVF from reservoir temperature, dissolved gas oil ratio, and specific gravity of both oil and dissolved gas. Firstly, structural risk minimization (SRM) principle of support vector regression (SVR) was employed to construct a robust model for estimating FVF from the aforementioned inputs. Subsequently, an alternating conditional expectation (ACE) was used for approximating optimal transformations of input/output data to a higher correlated data and consequently developing a sophisticated model between transformed data. Eventually, a committee machine with SVR and ACE was constructed through the use of hybrid genetic algorithm-pattern search (GA-PS). Committee machine integrates ACE and SVR models in an optimal linear combination such that makes benefit of both methods. A group of 342 data points was used for model development and a group of 219 data points was used for blind testing the constructed model. Results indicated that the committee machine performed better than individual models.

  1. Determination of Acetonitrile Volume Fraction in Mobile Phase by HPLC

    Institute of Scientific and Technical Information of China (English)

    WU Yi; WANG Zhi-wu; GU Jing-kai; WANG Ying-wu

    2008-01-01

    This paper reports the development and validation of an assay for the determination of acetonitrile in the recycled mobile phase using high performance liquid chromatography(HPLC).The method is based on that the retention in reversed-phase liquid chromatography increases with decreasing concentration of organic phase in the mobile phase.The natural logarithm of the capacity ratio for a given solute is linearly related to the volume fraction of the organic modifier in the mobile phase.For dimethylphthalate and diethylphthalate,the linearity range is 30%--60%,and for biphenyl and terphenyl,the range is 60%-95%.Precision values(RSD) were both <1% and the accuracy(RE) was in the range of ±1%.The assay was successfully applied to the determination of acetonitrile concentration of recycled mobile phase after the distillation of the column eluent in our laboratory.

  2. Volume regulation and shape bifurcation in the cell nucleus.

    Science.gov (United States)

    Kim, Dong-Hwee; Li, Bo; Si, Fangwei; Phillip, Jude M; Wirtz, Denis; Sun, Sean X

    2015-09-15

    Alterations in nuclear morphology are closely associated with essential cell functions, such as cell motility and polarization, and correlate with a wide range of human diseases, including cancer, muscular dystrophy, dilated cardiomyopathy and progeria. However, the mechanics and forces that shape the nucleus are not well understood. Here, we demonstrate that when an adherent cell is detached from its substratum, the nucleus undergoes a large volumetric reduction accompanied by a morphological transition from an almost smooth to a heavily folded surface. We develop a mathematical model that systematically analyzes the evolution of nuclear shape and volume. The analysis suggests that the pressure difference across the nuclear envelope, which is influenced by changes in cell volume and regulated by microtubules and actin filaments, is a major factor determining nuclear morphology. Our results show that physical and chemical properties of the extracellular microenvironment directly influence nuclear morphology and suggest that there is a direct link between the environment and gene regulation. © 2015. Published by The Company of Biologists Ltd.

  3. Cell volume homeostatic mechanisms: effectors and signalling pathways

    DEFF Research Database (Denmark)

    Hoffmann, E K; Pedersen, Stine Helene Falsig

    2011-01-01

    . Later work addressed the mechanisms through which cellular signalling pathways regulate the volume regulatory effectors or flux pathways. These studies were facilitated by the molecular identification of most of the relevant channels and transporters, and more recently also by the increased...... understanding of their structures. Finally, much current research in the field focuses on the most up- and downstream components of these paths: how cells sense changes in cell volume, and how cell volume changes in turn regulate cell function under physiological and pathophysiological conditions....

  4. Relation Between the Cell Volume and the Cell Cycle Dynamics in Mammalian cell

    Science.gov (United States)

    Magno, A. C. G.; Oliveira, I. L.; Hauck, J. V. S.

    2016-08-01

    The main goal of this work is to add and analyze an equation that represents the volume in a dynamical model of the mammalian cell cycle proposed by Gérard and Goldbeter (2011) [1]. The cell division occurs when the cyclinB/Cdkl complex is totally degraded (Tyson and Novak, 2011)[2] and it reaches a minimum value. At this point, the cell is divided into two newborn daughter cells and each one will contain the half of the cytoplasmic content of the mother cell. The equations of our base model are only valid if the cell volume, where the reactions occur, is constant. Whether the cell volume is not constant, that is, the rate of change of its volume with respect to time is explicitly taken into account in the mathematical model, then the equations of the original model are no longer valid. Therefore, every equations were modified from the mass conservation principle for considering a volume that changes with time. Through this approach, the cell volume affects all model variables. Two different dynamic simulation methods were accomplished: deterministic and stochastic. In the stochastic simulation, the volume affects every model's parameters which have molar unit, whereas in the deterministic one, it is incorporated into the differential equations. In deterministic simulation, the biochemical species may be in concentration units, while in stochastic simulation such species must be converted to number of molecules which are directly proportional to the cell volume. In an effort to understand the influence of the new equation a stability analysis was performed. This elucidates how the growth factor impacts the stability of the model's limit cycles. In conclusion, a more precise model, in comparison to the base model, was created for the cell cycle as it now takes into consideration the cell volume variation

  5. Cell Volume Regulation and Apoptotic Volume Decrease in Rat Distal Colon Superficial Enterocytes

    Directory of Open Access Journals (Sweden)

    Stefania Antico

    2013-12-01

    Full Text Available Background: The colon epithelium is physiologically exposed to osmotic stress, and the activation of cell volume regulation mechanisms is essential in colonocyte physiology. Moreover, colon is characterized by a high apoptotic rate of mature cells balancing the high division rate of stem cells. Aim: The aim of the present work was to investigate the main cell volume regulation mechanisms in rat colon surface colonocytes and their role in apoptosis. Methods: Cell volume changes were measured by light microscopy and video imaging on colon explants; apoptosis sign appearance was monitored by confocal microscopy on annexin V/propidium iodide labeled explants. Results: Superficial colonocytes showed a dynamic regulation of their cell volume during anisosmotic conditions with a Regulatory Volume Increase (RVI response following hypertonic shrinkage and Regulatory Volume Decrease (RVD response following hypotonic swelling. RVI was completely inhibited by bumetanide, while RVD was completely abolished by high K+ or iberiotoxin treatment and by extracellular Ca2+ removal. DIDS incubation was also able to affect the RVD response. When colon explants were exposed to H2O2 as apoptotic inducer, colonocytes underwent an isotonic volume decrease ascribable to Apoptotic Volume Decrease (AVD within about four hours of exposure. AVD was shown to precede annexin V positivity. It was also inhibited by high K+ or iberiotoxin treatment. Interestingly, treatment with iberiotoxin significantly inhibited apoptosis progression. Conclusions: In rat superficial colonocytes K+ efflux through high conductance Ca2+-activated K+ channels (BK channels was demonstrated to be the main mechanism of RVD and to plays also a crucial role in the AVD process and in the progression of apoptosis.

  6. Cell elasticity determines macrophage function.

    Directory of Open Access Journals (Sweden)

    Naimish R Patel

    Full Text Available Macrophages serve to maintain organ homeostasis in response to challenges from injury, inflammation, malignancy, particulate exposure, or infection. Until now, receptor ligation has been understood as being the central mechanism that regulates macrophage function. Using macrophages of different origins and species, we report that macrophage elasticity is a major determinant of innate macrophage function. Macrophage elasticity is modulated not only by classical biologic activators such as LPS and IFN-γ, but to an equal extent by substrate rigidity and substrate stretch. Macrophage elasticity is dependent upon actin polymerization and small rhoGTPase activation, but functional effects of elasticity are not predicted by examination of gene expression profiles alone. Taken together, these data demonstrate an unanticipated role for cell elasticity as a common pathway by which mechanical and biologic factors determine macrophage function.

  7. Accurately determining log and bark volumes of saw logs using high-resolution laser scan data

    Science.gov (United States)

    R. Edward Thomas; Neal D. Bennett

    2014-01-01

    Accurately determining the volume of logs and bark is crucial to estimating the total expected value recovery from a log. Knowing the correct size and volume of a log helps to determine which processing method, if any, should be used on a given log. However, applying volume estimation methods consistently can be difficult. Errors in log measurement and oddly shaped...

  8. Intracellular ion concentrations and cell volume during cholinergic stimulation of eccrine secretory coil cells

    Energy Technology Data Exchange (ETDEWEB)

    Takemura, T.; Sato, F.; Saga, K.; Suzuki, Y.; Sato, K. (Univ. of Iowa College of Medicine, Iowa City (USA))

    1991-02-01

    Methacholine (MCh)-induced changes in intracellular concentrations of Na, K, and Cl (( Na)i, (K)i, and (Cl)i, respectively) and in cellular dry mass (a measure of cell shrinkage) were examined in isolated monkey eccrine sweat secretory coils by electron probe X-ray microanalysis using the peripheral standard method. To further confirm the occurrence of cell shrinkage during MCh stimulation, the change in cell volume of dissociated clear and dark cells were directly determined under a light microscope equipped with differential interference contrast (DIC) optics. X-ray microanalysis revealed a biphasic increase in cellular dry mass in clear cells during continuous MCh stimulation; an initial increase of dry mass to 158% (of control) followed by a plateau at 140%, which correspond to the decrease in cell volume of 37 and 29%, respectively. The latter agrees with the MCh-induced cell shrinkage of 29% in dissociated clear cells. The MCh-induced increase in dry mass in myoepithelial cells was less than half that of clear cells. During the steady state of MCh stimulation, both (K+)i and (Cl)i of clear cells decreased by about 45%, whereas (Na)i increased in such a way to maintain the sum of (Na) i + (K)i constant. There was a small (12-15 mM) increase in (Na)i and a decrease in (K)i in myoepithelial cells during stimulation with MCh. Dissociated dark cells failed to significantly shrink during MCh stimulation. The decrease in (Cl)i in the face of constant (Na)i + (K)i suggests the accumulation of unknown anion(s) inside the clear cell during MCh stimulation.

  9. Cell volume regulation in epithelial physiology and cancer

    DEFF Research Database (Denmark)

    Pedersen, Stine Helene Falsig; Hoffmann, Else Kay; Novak, Ivana

    2013-01-01

    The physiological function of epithelia is transport of ions, nutrients, and fluid either in secretory or absorptive direction. All of these processes are closely related to cell volume changes, which are thus an integrated part of epithelial function. Transepithelial transport and cell volume...... transporters and channels with key physiological functions in epithelia and known roles in the development of cancer in these tissues. Their roles in cell survival, cell cycle progression, and development of drug resistance in epithelial cancers will be discussed....... such as cancer, transepithelial and cell volume regulatory ion transport are dys-regulated. Furthermore, epithelial architecture and coordinated ion transport function are lost, cell survival/death balance is altered, and new interactions with the stroma arise, all contributing to drug resistance. Since altered...

  10. Determination of a comfortable volume of mouthwash for rinsing

    NARCIS (Netherlands)

    Keukenmeester, R.S.; Slot, D.E.; Rosema, N.A.M.; van der Weijden, G.A.

    2012-01-01

    AIM: The purpose of this study was to assess patient comfort when rinsing for 30 s with 5, 10, 15, 20 or 30 ml volumes of mouthwash, with the goal of establishing the most agreeable volume. MATERIAL AND METHODS: The study was designed as a single-blind, clinical trial with duplicate assessments. Par

  11. 99mTc-albumin can replace 125I-albumin to determine plasma volume repeatedly

    DEFF Research Database (Denmark)

    Bonfils, Peter K; Damgaard, Morten; Stokholm, Knud H

    2012-01-01

    OBJECTIVE: Plasma volume assessment may be of importance in several disorders. The purpose of the present study was to compare the reliability of plasma volume measurements by technetium-labeled human serum albumin ((99m)Tc-HSA) with a simultaneously performed plasma volume determination with iod......OBJECTIVE: Plasma volume assessment may be of importance in several disorders. The purpose of the present study was to compare the reliability of plasma volume measurements by technetium-labeled human serum albumin ((99m)Tc-HSA) with a simultaneously performed plasma volume determination...

  12. Cell volume and membrane stretch independently control K+ channel activity

    DEFF Research Database (Denmark)

    Bomholtz, Sofia Hammami; Willumsen, Niels J; Olsen, Hervør L

    2009-01-01

    A number of potassium channels including members of the KCNQ family and the Ca(2+) activated IK and SK, but not BK, are strongly and reversibly regulated by small changes in cell volume. It has been argued that this general regulation is mediated through sensitivity to changes in membrane stretch...... was not affected by membrane stretch. The results indicate that (1) activation of BK channels by local membrane stretch is not mimicked by membrane stress induced by cell swelling, and (2) activation of KCNQ1 channels by cell volume increase is not mediated by local tension in the cell membrane. We conclude....... To test this hypothesis we have studied the regulation of KCNQ1 and BK channels after expression in Xenopus oocytes. Results from cell-attached patch clamp studies (approximately 50 microm(2) macropatches) in oocytes expressing BK channels demonstrate that the macroscopic volume-insensitive BK current...

  13. Determinants of Extraocular Muscle Volume in Patients with Graves' Disease

    Directory of Open Access Journals (Sweden)

    Samer El-Kaissi

    2012-01-01

    Full Text Available Background. To examine factors contributing to extraocular muscle (EOM volume enlargement in patients with Graves’ hyperthyroidism. Methods. EOM volumes were measured with orbital magnetic resonance imaging (MRI in 39 patients with recently diagnosed Graves’ disease, and compared to EOM volumes of 13 normal volunteers. Thyroid function tests, uptake on thyroid scintigraphy, anti-TSH-receptor antibody positivity and other parameters were then evaluated in patients with EOM enlargement. Results. 31/39 patients had one or more enlarged EOM, of whom only 2 patients had clinical EOM dysfunction. Compared to Graves’ disease patients with normal EOM volumes, those with EOM enlargement had significantly higher mean serum TSH (0.020±0.005 versus 0.007±0.002 mIU/L; P value 0.012, free-T4 (52.9±3.3 versus 41.2±1.7 pmol/L; P value 0.003 and technetium uptake on thyroid scintigraphy (13.51±1.7% versus 8.55±1.6%; P value 0.045. There were no differences between the 2 groups in anti-TSH-receptor antibody positivity, the proportion of males, tobacco smokers, or those with active ophthalmopathy. Conclusions. Patients with recently diagnosed Graves’ disease and EOM volume enlargement have higher serum TSH and more severe hyperthyroidism than patients with normal EOM volumes, with no difference in anti-TSH-receptor antibody positivity between the two groups.

  14. Signal Transduction Involved in Cell Volume Regulation

    NARCIS (Netherlands)

    Th. van der Wijk (Thea)

    2001-01-01

    textabstract1.fammalian cells are surrounded by a selective permeable plasma membrane that allmvs the interior of the cell to differ in composition from the surrounding solution. The plasma membrane is formed by a bilayer of (phospho-) lipids and contains many different proteins. Hydrophobic molecul

  15. Effects of Lugol's iodine solution and formalin on cell volume of three bloom-forming dinoflagellates

    Science.gov (United States)

    Yang, Yang; Sun, Xiaoxia; Zhao, Yongfang

    2016-09-01

    Fixatives are traditionally used in marine ecosystem research. The bias introduced by fixatives on the dimensions of plankton cells may lead to an overestimation or underestimation of the carbon biomass. To determine the impact of traditional fixatives on dinoflagellates during short- and long-term fixation, we analyzed the degree of change in three bloom-forming dinoflagellates (Prorocentrum micans, Scrippsiella trochoidea and Noctiluca scintillans) brought about by Lugol's iodine solution (hereafter Lugol's) and formalin. The fixation effects were species-specific. P. micans cell volume showed no significant change following long-term preservation, and S. trochoidea swelled by approximately 8.06% in Lugol's and by 20.97% in formalin as a percentage of the live cell volume, respectively. N. scintillans shrank significantly in both fixatives. The volume change due to formalin in N. scintillans was not concentration-dependent, whereas the volume shrinkage of N. scintillans cells fixed with Lugol's at a concentration of 2% was nearly six-fold that in cells fixed with Lugol's at a concentration of 0.6%-0.8%. To better estimate the volume of N. scintillans fixed in formalin at a concentration of 5%, we suggest that the conversion relationship was as follows: volume of live cell=volume of intact fixed cell/0.61. Apart from size change, damage induced by fixatives on N. scintillans was obvious. Lugol's is not a suitable fixative for N. scintillans due to high frequency of broken cells. Accurate carbon biomass estimate of N. scintillans should be performed on live samples. These findings help to improve the estimate of phytoplankton cell volume and carbon biomass in marine ecosystem.

  16. DETERMINANTS OF RED-BLOOD-CELL DEFORMABILITY IN RELATION TO CELL AGE

    NARCIS (Netherlands)

    BOSCH, FH; WERRE, JM; ROERDINKHOLDERSTOELWINDER, B; HULS, T; WILLEKENS, FLA; WICHERS, G; HALIE, MR

    1994-01-01

    Red blood cell (RBC) deformability was determined with an ektacytometer in fractions separated on the basis of differences in cell volume or density. Deformability was measured with ektacytometry (rpm-scan and osmo-scan). We studied three groups of RBC fractions:l. By counterflow centrifugation we o

  17. Sensors and signal transduction pathways in vertebrate cell volume regulation

    DEFF Research Database (Denmark)

    Hoffmann, Else K; Pedersen, Stine F

    2006-01-01

    will be discussed. In contrast to the simple pathway of osmosensing in yeast, cells from vertebrate organisms appear to exhibit multiple volume sensing systems, the specific mechanism(s) activated being cell type- and stimulus-dependent. Candidate sensors include integrins and growth factor receptors, while other...

  18. Bistability: requirements on cell-volume, protein diffusion, and thermodynamics.

    Directory of Open Access Journals (Sweden)

    Robert G Endres

    Full Text Available Bistability is considered wide-spread among bacteria and eukaryotic cells, useful, e.g., for enzyme induction, bet hedging, and epigenetic switching. However, this phenomenon has mostly been described with deterministic dynamic or well-mixed stochastic models. Here, we map known biological bistable systems onto the well-characterized biochemical Schlögl model, using analytical calculations and stochastic spatiotemporal simulations. In addition to network architecture and strong thermodynamic driving away from equilibrium, we show that bistability requires fine-tuning towards small cell volumes (or compartments and fast protein diffusion (well mixing. Bistability is thus fragile and hence may be restricted to small bacteria and eukaryotic nuclei, with switching triggered by volume changes during the cell cycle. For large volumes, single cells generally loose their ability for bistable switching and instead undergo a first-order phase transition.

  19. Determination of fiber volume in graphite/epoxy materials using computer image analysis

    Science.gov (United States)

    Viens, Michael J.

    1990-01-01

    The fiber volume of graphite/epoxy specimens was determined by analyzing optical images of cross sectioned specimens using image analysis software. Test specimens were mounted and polished using standard metallographic techniques and examined at 1000 times magnification. Fiber volume determined using the optical imaging agreed well with values determined using the standard acid digestion technique. The results were found to agree within 5 percent over a fiber volume range of 45 to 70 percent. The error observed is believed to arise from fiber volume variations within the graphite/epoxy panels themselves. The determination of ply orientation using image analysis techniques is also addressed.

  20. Cell volume and membrane stretch independently control K+ channel activity.

    Science.gov (United States)

    Hammami, Sofia; Willumsen, Niels J; Olsen, Hervør L; Morera, Francisco J; Latorre, Ramón; Klaerke, Dan A

    2009-05-15

    A number of potassium channels including members of the KCNQ family and the Ca(2+) activated IK and SK, but not BK, are strongly and reversibly regulated by small changes in cell volume. It has been argued that this general regulation is mediated through sensitivity to changes in membrane stretch. To test this hypothesis we have studied the regulation of KCNQ1 and BK channels after expression in Xenopus oocytes. Results from cell-attached patch clamp studies (approximately 50 microm(2) macropatches) in oocytes expressing BK channels demonstrate that the macroscopic volume-insensitive BK current increases with increasing negative hydrostatic pressure (suction) applied to the pipette. Thus, at a pipette pressure of -5.0 +/- 0.1 mmHg the increase amounted to 381 +/- 146% (mean +/- S.E.M., n = 6, P < 0.025). In contrast, in oocytes expressing the strongly volume-sensitive KCNQ1 channel, the current was not affected by membrane stretch. The results indicate that (1) activation of BK channels by local membrane stretch is not mimicked by membrane stress induced by cell swelling, and (2) activation of KCNQ1 channels by cell volume increase is not mediated by local tension in the cell membrane. We conclude that stretch and volume sensitivity can be considered two independent regulatory mechanisms.

  1. Volume regulatory hormones and plasma volume in pregnant women with sickle cell disorder

    Directory of Open Access Journals (Sweden)

    Bosede B Afolabi

    2016-09-01

    Full Text Available Background: Sickle cell disease (haemoglobin SS (HbSS mainly affects those of West African origin and is associated with hypervolaemia. Plasma volume rises by up to 50% in normal pregnancy but was previously found to be paradoxically contracted in late sickle cell pregnancy. The renin–angiotensin–aldosterone system is activated very early in human pregnancy to support the plasma volume expansion. We hypothesised that activation of the renin–angiotensin–aldosterone system would be blunted in pregnant women with sickle cell disease. Materials and methods: We measured plasma volume and concentrations of plasma renin, angiotensinogen, aldosterone and other volume-related hormones in a cross-sectional study of pregnant and non-pregnant Nigerian women with HbSS or HbAA. Results: Plasma volume was higher in non-pregnant HbSS than HbAA women, but had not risen by 16 weeks, unlike plasma volume in HbAA women. The concentration of plasma renin also rose significantly less by 16 weeks in HbSS; angiotensinogen and aldosterone concentrations increased. Conclusions: The lower plasma renin concentration at 16 weeks with HbSS could be either primary or secondary to vasoconstriction related to inadequate vasodilator activity. The contracted plasma volume might then stimulate aldosterone synthesis by non-angiotensin II dependent stimulation. Studies of vasodilators such as nitric oxide, vasodilator eicosanoids or the PlGF/VEGF/sFlT-1 axis in pregnant HbSS and HbAA women will test this hypothesis.

  2. Methods for determining enzymatic activity comprising heating and agitation of closed volumes

    Energy Technology Data Exchange (ETDEWEB)

    Thompson, David Neil; Henriksen, Emily DeCrescenzo; Reed, David William; Jensen, Jill Renee

    2016-03-15

    Methods for determining thermophilic enzymatic activity include heating a substrate solution in a plurality of closed volumes to a predetermined reaction temperature. Without opening the closed volumes, at least one enzyme is added, substantially simultaneously, to the closed volumes. At the predetermined reaction temperature, the closed volumes are agitated and then the activity of the at least one enzyme is determined. The methods are conducive for characterizing enzymes of high-temperature reactions, with insoluble substrates, with substrates and enzymes that do not readily intermix, and with low volumes of substrate and enzyme. Systems for characterizing the enzymes are also disclosed.

  3. Methods for determining enzymatic activity comprising heating and agitation of closed volumes

    Energy Technology Data Exchange (ETDEWEB)

    Thompson, David Neil; Henriksen, Emily DeCrescenzo; Reed, David William; Jensen, Jill Renee

    2016-03-15

    Methods for determining thermophilic enzymatic activity include heating a substrate solution in a plurality of closed volumes to a predetermined reaction temperature. Without opening the closed volumes, at least one enzyme is added, substantially simultaneously, to the closed volumes. At the predetermined reaction temperature, the closed volumes are agitated and then the activity of the at least one enzyme is determined. The methods are conducive for characterizing enzymes of high-temperature reactions, with insoluble substrates, with substrates and enzymes that do not readily intermix, and with low volumes of substrate and enzyme. Systems for characterizing the enzymes are also disclosed.

  4. Microfluidic Cell Volume Biosensor for High Throughput Drug Screening

    Science.gov (United States)

    2005-01-01

    method is non- invasive, and it provides real-time measurement of changes in cell volume for both adherent and suspended cells. The microfluidic nature of...channels as well as the release of organic osmolytes, such as taurine [11]. Upon returning to isotonic solution at the end of RVD we observed cell...Haussinger, Biochem. J, 313,697 (1996). [2] Sachs, F. Nature Structural Biology 9, 636-637 (2002). [3] F. Schliess, and D. Haussinger, Bio. Chem. 383

  5. Low Volume Resuscitation with Cell Impermeants

    Science.gov (United States)

    2014-10-01

    analysis of total tissue water (TTW) content by calculating [ wet -dry]/dry weight ratios. Dry weights were determined after drying the tissue slices in a 65...confirm whether the correct spelling should be “natriuresis.” [AQ8]: Please check and approve the inserted section head “CONCLUSIONS.” [AQ9]: Ref 1

  6. Cell volume regulation in epithelial physiology and cancer

    DEFF Research Database (Denmark)

    Pedersen, Stine Helene Falsig; Hoffmann, Else Kay; Novak, Ivana

    2013-01-01

    The physiological function of epithelia is transport of ions, nutrients, and fluid either in secretory or absorptive direction. All of these processes are closely related to cell volume changes, which are thus an integrated part of epithelial function. Transepithelial transport and cell volume...... expression of ion transporters and channels is now recognized as one of the hallmarks of cancer, it is timely to consider this especially for epithelia. Epithelial cells are highly proliferative and epithelial cancers, carcinomas, account for about 90% of all cancers. In this review we will focus on ion...... transporters and channels with key physiological functions in epithelia and known roles in the development of cancer in these tissues. Their roles in cell survival, cell cycle progression, and development of drug resistance in epithelial cancers will be discussed....

  7. Quantitative determination of volume on MR tomograms in communicating hydrocephalus

    Energy Technology Data Exchange (ETDEWEB)

    Langkowski, J.H.; Palmie, S.G.; Koschitzky, H. v.; Imme, M.; Maas, R.; Schmidt, K.H.; Heller, M.

    1989-02-01

    For patients with communicating hydrocephalus the implementation of a cardioventricular shunt is mostly an approved therapy. The proof of periventricular oedemas in addition to clinical signs seems to be a criterion for an indication for implantation and for the selection of a special valve. With the aid of a newly developed computer programme volume estimation of the ventricular system and the periventricular oedemas has been effected for six patients on T/sub 2/-weighted MR-tomograms (1.5 tesla). Quotients of the different volumina are correlated with cerebrospinal fluid (CSF) pressure obtained during shunt implantation or lumbar measurement. Distinct indications have been found for a correlation between the volume of the periventricular oedemas and the pressure measurements. (orig./GDG).

  8. Sickle cell patients are characterized by a reduced glycocalyx volume

    NARCIS (Netherlands)

    E.J. van Beers (Eduard); M. Nieuwdorp (Max); A.J. Duits (Ashley); L.M. Evers (Ludo); J.J.B. Schnog; B.J. Biemond (Bart)

    2008-01-01

    textabstractThe glycocalyx is an important anti-inflammatory and anti-adhesive barrier at the luminal side of endothelial cells. Glycocalyx volume was significantly reduced in sickle cell patients (HbSS/HbSβ0-thalassemia median 0.47L, IQR 0.27-0.66, HbSC/HbSβ+-thalassemia 0.23L, 0.0-0.58) compared w

  9. Transient receptor potential channels in mechanosensing and cell volume regulation

    DEFF Research Database (Denmark)

    Pedersen, Stine Falsig; Nilius, Bernd

    2007-01-01

    Transient receptor potential (TRP) channels are unique cellular sensors responding to a wide variety of extra- and intracellular signals, including mechanical and osmotic stress. In recent years, TRP channels from multiple subfamilies have been added to the list of mechano- and/or osmosensitive...... and involvement in cell volume regulation....

  10. KCNQ1 channels sense small changes in cell volume

    DEFF Research Database (Denmark)

    Grunnet, Morten; Jespersen, Thomas; MacAulay, Nanna

    2003-01-01

    . Incubation of the oocytes in cytochalasin D and experiments with truncated KCNQ1 channels suggest that KCNQ1 channels sense cell volume changes through interactions between the cytoskeleton and the N-terminus of the channel protein. From our results we propose that KCNQ1 and KCNQ4 channels play an important...

  11. Large volume high-pressure cell for inelastic neutron scattering

    Energy Technology Data Exchange (ETDEWEB)

    Wang, W.; Kamenev, K. V. [Centre for Science at Extreme Conditions and School of Engineering, University of Edinburgh, Edinburgh EH9 3JZ (United Kingdom); Sokolov, D. A.; Huxley, A. D. [SUPA, Centre for Science at Extreme Conditions and School of Physics and Astronomy, University of Edinburgh, Edinburgh EH9 3JZ (United Kingdom)

    2011-07-15

    Inelastic neutron scattering measurements typically require two orders of magnitude longer data collection times and larger sample sizes than neutron diffraction studies. Inelastic neutron scattering measurements on pressurised samples are particularly challenging since standard high-pressure apparatus restricts sample volume, attenuates the incident and scattered beams, and contributes background scattering. Here, we present the design of a large volume two-layered piston-cylinder pressure cell with optimised transmission for inelastic neutron scattering experiments. The design and the materials selected for the construction of the cell enable its safe use to a pressure of 1.8 GPa with a sample volume in excess of 400 mm{sup 3}. The design of the piston seal eliminates the need for a sample container, thus providing a larger sample volume and reduced absorption. The integrated electrical plug with a manganin pressure gauge offers an accurate measurement of pressure over the whole range of operational temperatures. The performance of the cell is demonstrated by an inelastic neutron scattering study of UGe{sub 2}.

  12. Large volume high-pressure cell for inelastic neutron scattering

    Science.gov (United States)

    Wang, W.; Sokolov, D. A.; Huxley, A. D.; Kamenev, K. V.

    2011-07-01

    Inelastic neutron scattering measurements typically require two orders of magnitude longer data collection times and larger sample sizes than neutron diffraction studies. Inelastic neutron scattering measurements on pressurised samples are particularly challenging since standard high-pressure apparatus restricts sample volume, attenuates the incident and scattered beams, and contributes background scattering. Here, we present the design of a large volume two-layered piston-cylinder pressure cell with optimised transmission for inelastic neutron scattering experiments. The design and the materials selected for the construction of the cell enable its safe use to a pressure of 1.8 GPa with a sample volume in excess of 400 mm3. The design of the piston seal eliminates the need for a sample container, thus providing a larger sample volume and reduced absorption. The integrated electrical plug with a manganin pressure gauge offers an accurate measurement of pressure over the whole range of operational temperatures. The performance of the cell is demonstrated by an inelastic neutron scattering study of UGe2.

  13. High-Volume Production of Lightweight Multijunction Solar Cells

    Science.gov (United States)

    Youtsey, Christopher

    2015-01-01

    MicroLink Devices, Inc., has transitioned its 6-inch epitaxial lift-off (ELO) solar cell fabrication process into a manufacturing platform capable of sustaining large-volume production. This Phase II project improves the ELO process by reducing cycle time and increasing the yield of large-area devices. In addition, all critical device fabrication processes have transitioned to 6-inch production tool sets designed for volume production. An emphasis on automated cassette-to-cassette and batch processes minimizes operator dependence and cell performance variability. MicroLink Devices established a pilot production line capable of at least 1,500 6-inch wafers per month at greater than 80 percent yield. The company also increased the yield and manufacturability of the 6-inch reclaim process, which is crucial to reducing the cost of the cells.

  14. Influence of skeletal muscle satellite cells implanted into infarcted myocardium on remnant myocyte volumes

    Institute of Scientific and Technical Information of China (English)

    钟竑; 朱洪生; 卫洪超; 张臻

    2003-01-01

    Objective To study the effects of skeletal muscle satellite cells implanted into infarcted myocardium on the volume of remnant myocytes.Methods Thirty-six adult mongrel canines were divided randomly into implantation group and control group. In the implantation group, skeletal muscle satellite cells taken from the gluteus maximus muscles of the dogs were cultured, proliferated and labeled with 4', 6-diamidino-2-phenylindone (DAPI) in vitro. In both groups, a model of acute myocardial infarction was established in every dog. In the implantation group, each dog was injected with M199 solution containing autologous skeletal muscle satellite cells. The dogs in the control group received M199 solution without skeletal muscle satellite cells. The dogs of both groups were killed 2, 4 and 8 weeks after implantation (six dogs in a separate group each time). Both infarcted myocardium and normal myocytes distal from the infracted regions isolated were observed under optical and fluorescent microscope. Their volumes were determined using a confocal microscopy image analysis system and analyzed using SAS. A P<0.05 was considered significant.Results A portion of the implanted cells differentiated into muscle fiber with striations and were connected with intercalated discs. Cross-sectional area and cell volume were increased in normal myocardium. Hypertrophy of remnant myocytes in the infarcted site after skeletal muscle cell implantation was much more evident than in the control group. Cross-sectional area, cell area and cell volume differed significantly from those of the control group (P< 0.05). Hypertrophy of the cells occurred predominantly in terms of width and thickness, whereas cell length remained unchanged. Conclusion Skeletal muscle satellite cells implanted into infarct myocardium, could induce the hypertrophy of remnant myocyte cells in the infarcted site and could also aid in the recovery of the contractile force of the infarcted myocardium.

  15. Technical Note: The determination of enclosed water volume in large flexible-wall mesocosms "KOSMOS"

    Directory of Open Access Journals (Sweden)

    J. Czerny

    2013-03-01

    Full Text Available The volume of water enclosed inside flexible-wall mesocosm bags is hard to estimate using geometrical calculations and can be strongly variable among bags of the same dimensions. Here we present a method for precise water volume determination in mesocosms using salinity as a tracer. Knowledge of the precise volume of water enclosed allows establishment of exactly planned treatment concentrations and calculation of elemental budgets.

  16. Determination of the column hold-up volume in supercritical fluid chromatography using nitrous-oxide.

    Science.gov (United States)

    Vajda, Péter; Guiochon, Georges

    2013-09-27

    This study introduces a new tracer that is useful for the determination of the hold-up time or volume of packed columns, particularly of those used in supercritical fluid chromatography. The thermodynamic void volume of three columns packed with different adsorbents were determined using the weight difference method. These void volumes were used as the reference point in the further discussion. The hold-up volumes of these columns were determined under dynamic conditions, using nitrous oxide dissolved in methanol as the hold-up time marker. Changes in the hold-up volumes of these columns were monitored during changes of the volumetric flow rate of pure supercritical carbon dioxide and of dilute mixtures of organic modifier and supercritical carbon dioxide. The results suggest significant methanol enrichment on the adsorbent surface.

  17. Effect of intraarticular osmic acid on synovial membrane volume and inflammation, determined by magnetic resonance imaging

    DEFF Research Database (Denmark)

    Østergaard, Mikkel; Stoltenberg, M; Gideon, P;

    1995-01-01

    The changes in MR-determined synovial membrane volume, early synovial enhancement, and cartilage and bone erosions after osmic acid knee synovectomy were studied. Gadolinium-DTPA enhanced magnetic resonance imaging (MRI) of 18 knees with persistent arthritis was performed before and 1 month after...... treatment. The synovial membrane volume was significantly reduced (median -52%) in all 9 patients brought into clinical remission (p

  18. Ultrasonographic determination of the thyroid volume in 7- 10 years old children of Bushehr port 2007

    Directory of Open Access Journals (Sweden)

    Farzad Morad Haseli

    2009-02-01

    Full Text Available Background: Determination of thyroid volume using ultrasound has been recommended for monitoring public control of iodine deficiency programs in areas with mild iodine deficiency or with no iodine deficiency, instead of physical examination, in order to estimate the prevalence rate of goiter. The aim of this study was to determine the reference ranges of thyroid volume in Bushehr port. Methods: Thyroid volume of 1247 primary schoolchildren aged 7-10 years who were selected by probability proportionate to size method from rural and urban areas of Bushehr port was determined using ultrasonography. Medians and percentiles of thyroid volumes for age and body surface area were calculated for both genders. Results: The 97th percentiles of thyroid volume of Bushehr port schoolchildren according to age and sex were all lower than the corresponding sex-specific normative WHO reference values. Goiter prevalence was 1.68% according to WHO thyroid volume references for age in the studied population. The goiter prevalence according to age and body surface area-specific new normative WHO reference values were 7.13 and 7.17%, respectively. Conclusion: The 97th percentile for thyroid volume, calculated by ultrasonography in Bushehr port schoolchildren, were lower than the international WHO newly recommended reference values in all ages. Therefore, determination of a native reference value for estimating the prevalence rate of thyroid is highly recommended. According to ultrasonographically determined goiter prevalence, Bushehr is an iodine sufficient area.

  19. Determining Outdoor CPV Cell Temperature (Presentation)

    Energy Technology Data Exchange (ETDEWEB)

    Muller, M.

    2011-04-01

    An accurate method is needed for determining cell temperature when measuring CPV modules outdoors. It has been suggested that cell temperature can be calculated though a procedure that shutters sunlight to the cells while measuring the transients in open-circuit voltage (Voc) and heat sink temperature. This presentation documents application of this shutter procedure to multiple CPV modules at NREL. The challenges and limitations are presented along with an alternate approach to measuring CPV cell operating temperature.

  20. Ion channels involved in cell volume regulation: effects on migration, proliferation, and programmed cell death in non adherent EAT cells and adherent ELA cells.

    Science.gov (United States)

    Hoffmann, Else Kay

    2011-01-01

    This mini review outlines studies of cell volume regulation in two closely related mammalian cell lines: nonadherent Ehrlich ascites tumour cells (EATC) and adherent Ehrlich Lettre ascites (ELA) cells. Focus is on the regulatory volume decrease (RVD) that occurs after cell swelling, the volume regulatory ion channels involved, and the mechanisms (cellular signalling pathways) that regulate these channels. Finally, I shall also briefly review current investigations in these two cell lines that focuses on how changes in cell volume can regulate cell functions such as cell migration, proliferation, and programmed cell death.

  1. Electroanalysis of metabolic flux from single cells in simple picoliter-volume microsystems.

    Science.gov (United States)

    Yasukawa, Tomoyuki; Glidle, Andrew; Cooper, Jonathan M; Matsue, Tomokazu

    2002-10-01

    A picoliter-volume electrochemical analytical chamber has been developed for detecting the metabolic flux resulting from the stress responses of a single plant cell. Electrochemical cells, with volumes as small as 100 pL, were fabricated by controlled electrochemical dissolution of a gold wire sealed in glass (the back-etching of the metal realizing an ultralow-volume titer chamber). In the first instance, the electrode contained within the chamber was characterized by the microinjection of standard aliquots of either ascorbic acid or hydrogen peroxide. In all cases, experimental currents obtained correlated well with theoretical calculations. Subsequently, single plant cells were micromanipulated into the chambers and were exposed to amounts of the detergent SDS (which permeabilized the cell membrane and released the intracellular contents). The flux of metabolite released from a single cell was estimated by using electrochemical-linked assays based upon the enzymes catalase, ascorbate oxidase, and horseradish peroxidase (in each case), in the presence of a mediator. In so doing, we investigated the activity of the cellular protection mechanisms through the determination of peroxides, while the individual cell was "stressed". The technique was found to provide a reliable and reproducible method for making single-cell measurements, using fabrication procedures that are both simple and do not require photolithographic methods.

  2. Activation of Salivary Secretion: Coupling of Cell Volume and [Ca2+]i in Single Cells

    Science.gov (United States)

    Foskett, J. Kevin; Melvin, James E.

    1989-06-01

    High-resolution differential interference contrast microscopy and digital imaging of the fluorescent calcium indicator dye fura-2 were performed simultaneously in single rat salivary gland acinar cells to examine the effects of muscarinic stimulation on cell volume and cytoplasmic calcium concentration ([Ca2+]i). Agonist stimulation of fluid secretion is initially associated with a rapid tenfold increase in [Ca2+]i as well as a substantial cell shrinkage. Subsequent changes of cell volume in the continued presence of agonist are tightly coupled to dynamic levels of [Ca2+]i, even during [Ca2+]i oscillations. Experiments with Ca2+ chelators and ionophores showed that physiological elevations of [Ca2+]i are necessary and sufficient to cause changes in cell volume. The relation between [Ca2+]i and cell volume suggests that the latter reflects the secretory state of the acinar cell. Agonist-induced changes in [Ca2+]i, by modulating specific ion permeabilities, result in solute movement into or out of the cell. The resultant cell volume changes may be important in modulating salivary secretion.

  3. Different approaches to synovial membrane volume determination by magnetic resonance imaging: manual versus automated segmentation

    DEFF Research Database (Denmark)

    Østergaard, Mikkel

    1997-01-01

    Automated fast (5-20 min) synovial membrane volume determination by MRI, based on pre-set post-gadolinium-DTPA enhancement thresholds, was evaluated as a substitute for a time-consuming (45-120 min), previously validated, manual segmentation method. Twenty-nine knees [rheumatoid arthritis (RA) 13...... or synovial membrane volume, e.g. no systematic errors were found. The inter-MRI variation, evaluated in three knees and three wrists, was higher than by manual segmentation, particularly due to sensitivity to malalignment artefacts. Examination of test objects proved the high accuracy of the general...... methodology for volume determinations (maximal error 6.3%). Preceded by the determination of reproducibility and the optimal threshold at the available MR unit, automated 'threshold' segmentation appears to be acceptable when changes rather than absolute values of synovial membrane volumes are most important...

  4. Comments on 'Ionization chamber volume determination and quality assurance using micro-CT imaging'.

    Science.gov (United States)

    Ross, C K

    2009-03-21

    The authors of a recent paper (McNiven et al 2008 Phys. Med. Biol. 53 5029-43) measured the volume of a particular type of a small ionization chamber using CT images. Using four Exradin A1SL chambers, they find that the volume measured using CT imaging is, on average, 4.3% larger than the value derived from the chamber calibration coefficient. Although they point out that the effective chamber volume is defined by electric field lines between the collector and the chamber body, they do not estimate how the mechanical volume might differ from the effective volume. We have used a commercial software package to calculate the electric field in the cavity and we show that the field lines define a volume that is about 11% smaller than the mechanical volume. We also show that the effective volume is very sensitive to small changes in the chamber geometry near the base of the collector. We conclude that simply determining the mechanical volume without careful consideration of the electric field lines within the cavity is not a useful dosimetric technique.

  5. Power law relationship between cell cycle duration and cell volume in the early embryonic development of Caenorhabditis elegans.

    Science.gov (United States)

    Arata, Yukinobu; Takagi, Hiroaki; Sako, Yasushi; Sawa, Hitoshi

    2014-01-01

    Cell size is a critical factor for cell cycle regulation. In Xenopus embryos after midblastula transition (MBT), the cell cycle duration elongates in a power law relationship with the cell radius squared. This correlation has been explained by the model that cell surface area is a candidate to determine cell cycle duration. However, it remains unknown whether this second power law is conserved in other animal embryos. Here, we found that the relationship between cell cycle duration and cell size in Caenorhabditis elegans embryos exhibited a power law distribution. Interestingly, the powers of the time-size relationship could be grouped into at least three classes: highly size-correlated, moderately size-correlated, and potentially a size-non-correlated class according to C. elegans founder cell lineages (1.2, 0.81, and power law relationship is conserved in Xenopus and C. elegans, while the absolute powers in C. elegans were different from that in Xenopus. Furthermore, we found that the volume ratio between the nucleus and cell exhibited a power law relationship in the size-correlated classes. The power of the volume relationship was closest to that of the time-size relationship in the highly size-correlated class. This correlation raised the possibility that the time-size relationship, at least in the highly size-correlated class, is explained by the volume ratio of nuclear size and cell size. Thus, our quantitative measurements shed a light on the possibility that early embryonic C. elegans cell cycle duration is coordinated with cell size as a result of geometric constraints between intracellular structures.

  6. Left ventricular volume determined from scintigraphy and digital angiography by a semi-automated geometric method

    Energy Technology Data Exchange (ETDEWEB)

    Seldin, D.W.; Esser, P.D.; Nichols, A.B.; Ratner, S.J.; Alderson, P.O.

    1983-12-01

    The utility of a semi-automatic method of measuring left ventricular (LV) volume geometrically from gated blood-pool studies and digital subtraction angiography (DSA) was investigated using computerized edge detection and spatial calibration algorithms. LAO LV volumes determined from gated blood-pool studies were compared to volumes obtained from contrast left ventriculograms in 21 patients and the applicability of this method to DSA was evaluated in 25 additional patients who also had conventional left ventriculography. There was excellent correlation between the two, both for radionuclide studies and for DSA. Computer-based geometric determinations of LV volume appear to be rapid, accurate, and less dependent on subjective operator decisions than previously reported geometric approaches.

  7. 40 CFR Appendix D to Part 112 - Determination of a Worst Case Discharge Planning Volume

    Science.gov (United States)

    2010-07-01

    ... 40 Protection of Environment 21 2010-07-01 2010-07-01 false Determination of a Worst Case Discharge Planning Volume D Appendix D to Part 112 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) WATER PROGRAMS OIL POLLUTION PREVENTION Pt. 112, App. D Appendix D to Part 112—Determination of...

  8. Determination of equine deep digital flexor muscle volume based on distances between anatomical landmarks

    NARCIS (Netherlands)

    Hardeman, L C; van der Meij, B R; Lamers, A A H; van der Kolk, J H; Back, W; Wijnberg, I D

    2014-01-01

    In equine medicine the use of Botox® is experimental. Dosages are determined from human treatment-protocols and limited numbers of equine studies. Determination of target-muscle volume can be helpful to extrapolate human dosages. The aim of the study was to calculate a formula enabling the

  9. Evaluation of a simple method for determining muscle volume in vivo.

    Science.gov (United States)

    Infantolino, Benjamin W; Challis, John H

    2016-06-14

    The quantification in vivo of muscle volume is important, for example, to understand how muscles change with aging, and respond to rehabilitation. Albracht et al. (2008) suggested that muscle volume can be estimated in vivo from the measurement of muscle cross-sectional area and muscle belly length only. The purpose of this study was to evaluate this proposed relationship for determining muscle volume for both the Vastus Lateralis (VL) and First Dorsal Interosseous (FDI) using ultrasound imaging. The cross-sectional area and length of 22 cadaver FDI and 6 VL muscles in cadavers were imaged using ultrasound, these muscles were then dissected and muscle volumes measured directly using the water displacement technique. Estimated muscle volumes were compared with their direct measurement, and for the VL the percentage root mean square error in the estimation of muscle volume was 5.0%, and the Bland-Altman analysis had all volume estimates within the 95% confidence interval, with no evidence of bias (proportional or constant) in the volume estimates. In contrast, percentage root mean square error for the FDI was 18.8%, with the Bland-Altman analysis showing volume estimates outside of the 95% confidence interval and proportional bias. These results indicate that the simple method proposed by Albracht et al. (2008) for the estimation of muscle volume is appropriate the VL but not the FDI using ultrasound imaging. Morphological disparities likely account for these differences, if accurate and fast measures of the volume of the FDI are required other approaches should be explored.

  10. Left ventricular volume determination from single-photon emission computed tomography

    Energy Technology Data Exchange (ETDEWEB)

    Bunker, S.R.; Hartshorne, M.F.; Schmidt, W.P.; Cawthon, M.A.; Karl, R.D. Jr.; Bauman, J.M.; Howard, W.H. III; Rubal, B.J.

    1985-02-01

    To compare the accuracy of single-photon emission computed tomography (SPECT) with that of contrast cineangiography in measuring left ventricular end-diastolic volume, 25 consecutive patients undergoing catheterizaiton for coronary artery or valvular heart disease were first evaluated scintigraphically. SPECT volume values showed a high degree of correlation with those determined by angiography with a standard error of the estimate of 23 ml. SPECT offers a highly accurate and essentially noninvasive method for measuring chamber volumes that is independent of geometric assumptions about ventricular configuration and chest wall attenuation and does not require blood sample counting.

  11. Determinative developmental cell lineages are robust to cell deaths.

    Directory of Open Access Journals (Sweden)

    Jian-Rong Yang

    2014-07-01

    Full Text Available All forms of life are confronted with environmental and genetic perturbations, making phenotypic robustness an important characteristic of life. Although development has long been viewed as a key component of phenotypic robustness, the underlying mechanism is unclear. Here we report that the determinative developmental cell lineages of two protostomes and one deuterostome are structured such that the resulting cellular compositions of the organisms are only modestly affected by cell deaths. Several features of the cell lineages, including their shallowness, topology, early ontogenic appearances of rare cells, and non-clonality of most cell types, underlie the robustness. Simple simulations of cell lineage evolution demonstrate the possibility that the observed robustness arose as an adaptation in the face of random cell deaths in development. These results reveal general organizing principles of determinative developmental cell lineages and a conceptually new mechanism of phenotypic robustness, both of which have important implications for development and evolution.

  12. DETERMINATION OF THE AGR-1 CAPSULE TO FPMS SPECTROMETER TRANSPORT VOLUMES FROM LEADOUT FLOW TEST DATA

    Energy Technology Data Exchange (ETDEWEB)

    J. K. Hartwell; J. B. Walter; D. M. Scates; M. W. Drigert

    2007-05-01

    The AGR-1 experiment is a fueled multiple-capsule irradiation experiment being conducted in the Advanced Test Reactor (ATR) in support of the Advanced Gas Reactor (AGR) Fuel Development and Qualification Program. A flow experiment conducted during the AGR-1 irradiation provided data that included the effect of flow rate changes on the decay of a short-lived radionuclide (23Ne). This data has been analyzed to determine the capsule-specific downstream transport volume through which the capsule effluents must pass before arrival at the fission product monitoring system spectrometers. These resultant transport volumes when coupled with capsule outlet flow rates determine the transport times from capsule-to-detector. In this work an analysis protocol is developed and applied in order to determine capsule-specific transport volumes to precisions of better than +/- 7%.

  13. Aquaporin 2-increased renal cell proliferation is associated with cell volume regulation.

    Science.gov (United States)

    Di Giusto, Gisela; Flamenco, Pilar; Rivarola, Valeria; Fernández, Juan; Melamud, Luciana; Ford, Paula; Capurro, Claudia

    2012-12-01

    We have previously demonstrated that in renal cortical collecting duct cells (RCCD(1)) the expression of the water channel Aquaporin 2 (AQP2) raises the rate of cell proliferation. In this study, we investigated the mechanisms involved in this process, focusing on the putative link between AQP2 expression, cell volume changes, and regulatory volume decrease activity (RVD). Two renal cell lines were used: WT-RCCD(1) (not expressing aquaporins) and AQP2-RCCD(1) (transfected with AQP2). Our results showed that when most RCCD(1) cells are in the G(1)-phase (unsynchronized), the blockage of barium-sensitive K(+) channels implicated in rapid RVD inhibits cell proliferation only in AQP2-RCCD(1) cells. Though cells in the S-phase (synchronized) had a remarkable increase in size, this enhancement was higher and was accompanied by a significant down-regulation in the rapid RVD response only in AQP2-RCCD(1) cells. This decrease in the RVD activity did not correlate with changes in AQP2 function or expression, demonstrating that AQP2-besides increasing water permeability-would play some other role. These observations together with evidence implying a cell-sizing mechanism that shortens the cell cycle of large cells, let us to propose that during nutrient uptake, in early G(1), volume tends to increase but it may be efficiently regulated by an AQP2-dependent mechanism, inducing the rapid activation of RVD channels. This mechanism would be down-regulated when volume needs to be increased in order to proceed into the S-phase. Therefore, during cell cycle, a coordinated modulation of the RVD activity may contribute to accelerate proliferation of cells expressing AQP2. Copyright © 2012 Wiley Periodicals, Inc.

  14. Determination of air-loop volume and radon partition coefficient for measuring radon in water sample.

    Science.gov (United States)

    Lee, Kil Yong; Burnett, William C

    A simple method for the direct determination of the air-loop volume in a RAD7 system as well as the radon partition coefficient was developed allowing for an accurate measurement of the radon activity in any type of water. The air-loop volume may be measured directly using an external radon source and an empty bottle with a precisely measured volume. The partition coefficient and activity of radon in the water sample may then be determined via the RAD7 using the determined air-loop volume. Activity ratios instead of absolute activities were used to measure the air-loop volume and the radon partition coefficient. In order to verify this approach, we measured the radon partition coefficient in deionized water in the temperature range of 10-30 °C and compared the values to those calculated from the well-known Weigel equation. The results were within 5 % variance throughout the temperature range. We also applied the approach for measurement of the radon partition coefficient in synthetic saline water (0-75 ppt salinity) as well as tap water. The radon activity of the tap water sample was determined by this method as well as the standard RAD-H2O and BigBottle RAD-H2O. The results have shown good agreement between this method and the standard methods.

  15. A simple, quantitative method using alginate gel to determine rat colonic tumor volume in vivo.

    Science.gov (United States)

    Irving, Amy A; Young, Lindsay B; Pleiman, Jennifer K; Konrath, Michael J; Marzella, Blake; Nonte, Michael; Cacciatore, Justin; Ford, Madeline R; Clipson, Linda; Amos-Landgraf, James M; Dove, William F

    2014-04-01

    Many studies of the response of colonic tumors to therapeutics use tumor multiplicity as the endpoint to determine the effectiveness of the agent. These studies can be greatly enhanced by accurate measurements of tumor volume. Here we present a quantitative method to easily and accurately determine colonic tumor volume. This approach uses a biocompatible alginate to create a negative mold of a tumor-bearing colon; this mold is then used to make positive casts of dental stone that replicate the shape of each original tumor. The weight of the dental stone cast correlates highly with the weight of the dissected tumors. After refinement of the technique, overall error in tumor volume was 16.9% ± 7.9% and includes error from both the alginate and dental stone procedures. Because this technique is limited to molding of tumors in the colon, we utilized the Apc(Pirc/+) rat, which has a propensity for developing colonic tumors that reflect the location of the majority of human intestinal tumors. We have successfully used the described method to determine tumor volumes ranging from 4 to 196 mm³. Alginate molding combined with dental stone casting is a facile method for determining tumor volume in vivo without costly equipment or knowledge of analytic software. This broadly accessible method creates the opportunity to objectively study colonic tumors over time in living animals in conjunction with other experiments and without transferring animals from the facility where they are maintained.

  16. Temporal dynamics and determinants of whole brain tissue volume changes during recovery from alcohol dependence.

    Science.gov (United States)

    Gazdzinski, Stefan; Durazzo, Timothy C; Meyerhoff, Dieter J

    2005-06-01

    Brain shrinkage and its partial reversibility with abstinence is a common neuroimaging finding in alcohol dependent individuals. We used an automated three-dimensional whole brain magnetic resonance imaging method (boundary shift integral) in 23 alcohol dependent individuals to measure the temporal dynamics of cerebral tissue and spinal fluid volume changes over a 12-month interval and to examine the major determinants of brain tissue change rates during abstinence and non-abstinence. We found more rapid brain tissue gain during the first month of sobriety than in the following months. The most rapid volume recovery was observed in abstinent individuals with the greatest baseline brain shrinkage and drinking severity. The rapid reversal of brain volume gains in non-abstinent individuals and tissue volume changes are modulated by duration of abstinence and non-abstinence periods, as well as recency of non-abstinence. Age, family history density of alcoholism, relapse severity, and duration or age of onset of heavy drinking were not major determinants of brain shrinkage and brain volume recovery rates. Treatment providers may use this tangible information to reinforce the biomedical benefits of sobriety. Previous quantitative measurements of brain volumes in alcohol dependent individuals performed after several weeks of abstinence likely underestimated the full extent of chronic alcohol-associated brain shrinkage.

  17. Method and system for determining a volume of an object from two-dimensional images

    Science.gov (United States)

    Abercrombie, Robert K [Knoxville, TN; Schlicher, Bob G [Portsmouth, NH

    2010-08-10

    The invention provides a method and a computer program stored in a tangible medium for automatically determining a volume of three-dimensional objects represented in two-dimensional images, by acquiring at two least two-dimensional digitized images, by analyzing the two-dimensional images to identify reference points and geometric patterns, by determining distances between the reference points and the component objects utilizing reference data provided for the three-dimensional object, and by calculating a volume for the three-dimensional object.

  18. Influence of flotation cell volume and solids mass on kinetics of sulfide ore flotation

    OpenAIRE

    Plawski Michal; Bakalarz Alicja

    2016-01-01

    The paper presents studies on the influence of flotation cell capacity and mass of solids in the suspension on the flotation kinetics of sulfide copper ore. A sample of copper ore that was collected from the Polkowice Mine of KGHM Polska Miedz S.A. in Poland was used in the experiments. It was determined that neither the volume of flotation cell nor the mass of solids had influence on the type of kinetics equation of flotation. Copper-bearing minerals floated according to the second-order equ...

  19. Clinical value of prostate segmentation and volume determination on MRI in benign prostatic hyperplasia.

    Science.gov (United States)

    Garvey, Brian; Türkbey, Barış; Truong, Hong; Bernardo, Marcelino; Periaswamy, Senthil; Choyke, Peter L

    2014-01-01

    Benign prostatic hyperplasia (BPH) is a nonmalignant pathological enlargement of the prostate, which occurs primarily in the transitional zone. BPH is highly prevalent and is a major cause of lower urinary tract symptoms in aging males, although there is no direct relationship between prostate volume and symptom severity. The progression of BPH can be quantified by measuring the volumes of the whole prostate and its zones, based on image segmentation on magnetic resonance imaging. Prostate volume determination via segmentation is a useful measure for patients undergoing therapy for BPH. However, prostate segmentation is not widely used due to the excessive time required for even experts to manually map the margins of the prostate. Here, we review and compare new methods of prostate volume segmentation using both manual and automated methods, including the ellipsoid formula, manual planimetry, and semiautomated and fully automated segmentation approaches. We highlight the utility of prostate segmentation in the clinical context of assessing BPH.

  20. Cell volume regulation in cultured human retinal Muller cells is associated with changes in transmembrane potential.

    Directory of Open Access Journals (Sweden)

    Juan M Fernández

    Full Text Available Müller cells are mainly involved in controlling extracellular homeostasis in the retina, where intense neural activity alters ion concentrations and osmotic gradients, thus favoring cell swelling. This increase in cell volume is followed by a regulatory volume decrease response (RVD, which is known to be partially mediated by the activation of K(+ and anion channels. However, the precise mechanisms underlying osmotic swelling and subsequent cell volume regulation in Müller cells have been evaluated by only a few studies. Although the activation of ion channels during the RVD response may alter transmembrane potential (Vm, no studies have actually addressed this issue in Müller cells. The aim of the present work is to evaluate RVD using a retinal Müller cell line (MIO-M1 under different extracellular ionic conditions, and to study a possible association between RVD and changes in Vm. Cell volume and Vm changes were evaluated using fluorescent probe techniques and a mathematical model. Results show that cell swelling and subsequent RVD were accompanied by Vm depolarization followed by repolarization. This response depended on the composition of extracellular media. Cells exposed to a hypoosmotic solution with reduced ionic strength underwent maximum RVD and had a larger repolarization. Both of these responses were reduced by K(+ or Cl(- channel blockers. In contrast, cells facing a hypoosmotic solution with the same ionic strength as the isoosmotic solution showed a lower RVD and a smaller repolarization and were not affected by blockers. Together, experimental and simulated data led us to propose that the efficiency of the RVD process in Müller glia depends not only on the activation of ion channels, but is also strongly modulated by concurrent changes in the membrane potential. The relationship between ionic fluxes, changes in ion permeabilities and ion concentrations -all leading to changes in Vm- define the success of RVD.

  1. Computer aided classification of cell nuclei in the gastrointestinal tract by volume and principal axis

    Science.gov (United States)

    Sagstetter, Ann M.; Camp, Jon J.; Lurken, Matthew S.; Szurszewski, Joseph H.; Farrugia, Gianrico; Gibbons, Simon J.; Robb, Richard A.

    2007-03-01

    Normal function of the gastrointestinal tract involves the coordinated activity of several cell types Human disorders of motor function of the gastrointestinal tract are often associated with changes in the number of these cells. For example, in diabetic patients, abnormalities in gastrointestinal transit are associated with changes in nerves and interstitial cells of Cajal (ICC), two key cells that generate and regulate motility. ICC are cells of mesenchymal origin that function as pacemakers and amplify neuronal signals in the gastrointestinal tract. Quantifying the changes in number of specific cell types in tissues from patients with motility disorders is challenging and requires immunolabeling for specific antigens. The shape of nuclei differs between the cell types in the wall of the gastrointestinal tract. Therefore the objective of this study was to determine whether cell nuclei can be classified by analyzing the 3D morphology of the nuclei. Furthermore, the orientation of the long axis of nuclei changes within and between the muscle layers. These features can be used to classify and differentially label the nuclei in confocal volume images of the tissue by computing the principal axis of the coordinates of the set of voxels forming each nucleus and thereby to identify cells by their nuclear morphology. Using this approach, we were able to separate and quantify nuclei in the smooth muscle layers of the tissue. Therefore we conclude that computer-aided classification of cell nuclei can be used to identify changes in the cell types expressed in gastrointestinal smooth muscle.

  2. Determining physical properties of the cell cortex

    CERN Document Server

    Saha, A; Behrndt, M; Heisenberg, C -P; Jülicher, F; Grill, S W

    2015-01-01

    Actin and myosin assemble into a thin layer of a highly dynamic network underneath the membrane of eukaryotic cells. This network generates the forces that drive cell and tissue-scale morphogenetic processes. The effective material properties of this active network determine large-scale deformations and other morphogenetic events. For example,the characteristic time of stress relaxation (the Maxwell time)in the actomyosin sets the time scale of large-scale deformation of the cortex. Similarly, the characteristic length of stress propagation (the hydrodynamic length) sets the length scale of slow deformations, and a large hydrodynamic length is a prerequisite for long-ranged cortical flows. Here we introduce a method to determine physical parameters of the actomyosin cortical layer (in vivo). For this we investigate the relaxation dynamics of the cortex in response to laser ablation in the one-cell-stage {\\it C. elegans} embryo and in the gastrulating zebrafish embryo. These responses can be interpreted using ...

  3. Effect of intraarticular osmic acid on synovial membrane volume and inflammation, determined by magnetic resonance imaging

    DEFF Research Database (Denmark)

    Østergaard, Mikkel; Stoltenberg, M; Gideon, P

    1995-01-01

    The changes in MR-determined synovial membrane volume, early synovial enhancement, and cartilage and bone erosions after osmic acid knee synovectomy were studied. Gadolinium-DTPA enhanced magnetic resonance imaging (MRI) of 18 knees with persistent arthritis was performed before and 1 month after...

  4. Tutorial for Collecting and Processing Images of Composite Structures to Determine the Fiber Volume Fraction

    Science.gov (United States)

    Conklin, Lindsey

    2017-01-01

    Fiber-reinforced composite structures have become more common in aerospace components due to their light weight and structural efficiency. In general, the strength and stiffness of a composite structure are directly related to the fiber volume fraction, which is defined as the fraction of fiber volume to total volume of the composite. The most common method to measure the fiber volume fraction is acid digestion, which is a useful method when the total weight of the composite, the fiber weight, and the total weight can easily be obtained. However, acid digestion is a destructive test, so the material will no longer be available for additional characterization. Acid digestion can also be difficult to machine out specific components of a composite structure with complex geometries. These disadvantages of acid digestion led the author to develop a method to calculate the fiber volume fraction. The developed method uses optical microscopy to calculate the fiber area fraction based on images of the cross section of the composite. The fiber area fraction and fiber volume fraction are understood to be the same, based on the assumption that the shape and size of the fibers are consistent in the depth of the composite. This tutorial explains the developed method for optically determining fiber area fraction performed at NASA Langley Research Center.

  5. Closed system generation of dendritic cells from a single blood volume for clinical application in immunotherapy.

    Science.gov (United States)

    Elias, M; van Zanten, J; Hospers, G A P; Setroikromo, A; de Jong, M A; de Leij, L F M H; Mulder, N H

    2005-12-01

    Dendritic cells (DC) used for clinical trials should be processed on a large scale conforming to current good manufacturing practice (cGMP) guidelines. The aim of this study was to develop a protocol for clinical grade generation of immature DC in a closed-system. Aphereses were performed with the Cobe Spectra continuous flow cell separator and material was derived from one volume of blood processed. Optimisation of a 3-phase collection autoPBSC technique significantly improved the quality of the initial mononuclear cell (MNC) product. Monocytes were then enriched from MNC by immunomagnetic depletion of CD19+ B cells and CD2+ T cells and partial depletion of NK cells using the Isolex 300I Magnetic cell selector. The quality of the initial mononuclear cell product was found to determine the outcome of monocyte enrichment. Enriched monocytes were cultured in Opticyte gas-permeable containers using CellGro serum-free medium supplemented with GM-CSF and IL-4 to generate immature DC. A seeding concentration of 1 x 10(6) was found optimal in terms of DC phenotype expression, monocyte percentage in culture, and cell viability. The differentiation pattern favours day 7 for harvest of immature DC. DC recovery, viability, as well as phenotype expression after cryopreservation of immature DC was considered in this study. DC were induced to maturation and evaluated in FACS analysis for phenotype expression and proliferation assays. Mature DC were able to generate an allogeneic T-cell response as well as an anti-CMV response as detected by proliferation assays. These data indicate that the described large-scale GMP-compatible system results in the generation of stable DC derived from one volume of blood processed, which are qualitatively and quantitatively sufficient for clinical application in immunotherapeutic protocols.

  6. Influence of flotation cell volume and solids mass on kinetics of sulfide ore flotation

    Directory of Open Access Journals (Sweden)

    Plawski Michal

    2016-01-01

    Full Text Available The paper presents studies on the influence of flotation cell capacity and mass of solids in the suspension on the flotation kinetics of sulfide copper ore. A sample of copper ore that was collected from the Polkowice Mine of KGHM Polska Miedz S.A. in Poland was used in the experiments. It was determined that neither the volume of flotation cell nor the mass of solids had influence on the type of kinetics equation of flotation. Copper-bearing minerals floated according to the second-order equation, while the remaining components according to the first-order equation. The kinetic rate constants and maximum recovery of the studied components decreased with increasing solids mass in the flotation cell, regardless of the capacity of the cell. The best results were obtained for tests using a 1.0 dm3 cell, while the less favorable kinetics results were observed in the test with the smallest cell of 0.75 dm3 volume. The obtained results can be helpful in choosing the most appropriate methodology of upgrading the sulfide copper ore from Poland in order to obtain the best kinetics results.

  7. Diffusion on a curved surface coupled to diffusion in the volume: Application to cell biology

    Science.gov (United States)

    Novak, Igor L.; Gao, Fei; Choi, Yung-Sze; Resasco, Diana; Schaff, James C.; Slepchenko, Boris M.

    2007-10-01

    An algorithm is presented for solving a diffusion equation on a curved surface coupled to diffusion in the volume, a problem often arising in cell biology. It applies to pixilated surfaces obtained from experimental images and performs at low computational cost. In the method, the Laplace-Beltrami operator is approximated locally by the Laplacian on the tangential plane and then a finite volume discretization scheme based on a Voronoi decomposition is applied. Convergence studies show that mass conservation built in the discretization scheme and cancellation of sampling error ensure convergence of the solution in space with an order between 1 and 2. The method is applied to a cell-biological problem where a signaling molecule, G-protein Rac, cycles between the cytoplasm and cell membrane thus coupling its diffusion in the membrane to that in the cell interior. Simulations on realistic cell geometry are performed to validate, and determine the accuracy of, a recently proposed simplified quantitative analysis of fluorescence loss in photobleaching. The method is implemented within the Virtual Cell computational framework freely accessible at http://www.vcell.org.

  8. Functional Response of Tumor Vasculature to PaCO2: Determination of Total and Microvascular Blood Volume by MRI

    Directory of Open Access Journals (Sweden)

    Scott D. Packard

    2003-07-01

    Full Text Available In order to identify differences in functional activity, we compared the reactivity of glioma vasculature and the native cerebral vasculature to both dilate and constrict in response to altered PaCO2. Gliomas were generated by unilateral implantation of U87MGdEGFR human glioma tumor cells into the striatum of adult female athymic rats. Relative changes in total and microvascular cerebral blood volume were determined by steady state contrast agent-enhanced magnetic resonance imaging for transitions from normocarbia to hypercarbia and hypocarbia. Although hypercarbia induced a significant increase in both total and microvascular blood volume in normal brain and glioma, reactivity of glioma vasculature was significantly blunted in comparison to normal striatum; glioma total CBV increased by 0.6±0.1%/mm Hg CO2 whereas normal striatum increased by 1.5±0.2%/mm Hg CO2, (P < .0001, group t-test. Reactivity of microvascular blood volume was also significantly blunted. In contrast, hypocarbia decreased both total and microvascular blood volumes more in glioma than in normal striatum. These results indicate that cerebral blood vessels derived by tumor-directed angiogenesis do retain reactivity to CO2. Furthermore, reduced reactivity of tumor vessels to a single physiological perturbation, such as hypercarbia, should not be construed as a generalized reduction of functional activity of the tumor vascular bed.

  9. Tumor volume in subcutaneous mouse xenografts measured by microCT is more accurate and reproducible than determined by 18F-FDG-microPET or external caliper

    DEFF Research Database (Denmark)

    Jensen, Mette Munk; Jørgensen, Jesper Tranekjaer; Binderup, Tina;

    2008-01-01

    and reproducible measures of tumor size in mice compared with caliper measurements. Furthermore, we evaluated the accuracy of tumor volume determined from 18F-fluorodeoxyglucose (18F-FDG) PET. METHODS: Subcutaneously implanted human breast adenocarcinoma cells in NMRI nude mice served as tumor model. Tumor volume...... systematic bias compared to reference volume. Coefficients of variation for intra-observer variation were 7% and 14% for microCT and caliper measurements, respectively. Regression coefficients between observers were 0.97 for microCT and 0.91 for caliper measurements. CONCLUSION: MicroCT was more accurate...

  10. Determination of Partial Molar Volumes of EPA and DHA Ethyl Esters in Supercritical Carbon Dioxide

    Institute of Scientific and Technical Information of China (English)

    2002-01-01

    The use of supercritical-fluid chromatography for determining partial molar volumes of ethyl esters of cis-5,8,11,14,17-eicosapentaenoic acid (EPA) and cis -4,7,10,13,16,19- docosa-hexaenoic acid (DHA) in supercritical carbon dioxide is presented and discussed. Partial molar volumes of EPA and DHA esters are obtained from the variation of the retention properties with the density of mobile phase at 313.15 K, 323.15 K, 333.15 K and in the pressure range from 9 MPa to 21 MPa.

  11. Regulation of Cytoplasmic and Vacuolar Volumes by Plant Cells in Suspension Culture

    DEFF Research Database (Denmark)

    Owens, Trevor; Poole, Ronald J

    1979-01-01

    Quantitative microscopical measurements have been made of the proportion of cell volume occupied by cytoplasm in a cell suspension culture derived from cotyledons of bush bean (cv. Contender). On a 7-day culture cycle, the content of cytoplasm varies from 25% at the time of transfer to 45......% at the start of the phase of rapid cell division. If the culture is continued beyond 7 days, the vacuole volume reaches 90% of cell volume by day 12.Attempts to measure relative cytoplasmic volumes by compartmental analysis of nonelectrolyte efflux were unsuccessful. The proportion of cell volume occupied...... by cytoplasm is roughly correlated with protein content, but shows no correlation with cell size or with intracellular concentrations of K or Na. The most striking observation is that the growth of cytoplasmic volume for the culture as a whole appears to be constant throughout the culture cycle, despite...

  12. The determinants of fuel use in the trucking industry – volume, size and the rebound effect

    DEFF Research Database (Denmark)

    Mulalic, Ismir

    2011-01-01

    We analyse the determinants of trucking firm fuel use. We develop a simple model to show that trucking firm fuel use depends, in addition to the fuel price and the traffic volume, also on the output of the trucking firm’s production process (the movement of cargo) measured in tonkilometres...... decompose the standard definition of the rebound effect for motor vehicles, i.e. the elasticity of traffic volume with respect to fuel cost, into the elasticity by which changes in fuel costs affects freight activity and the elasticity by which changes in freight activity affect traffic volume. We estimate...... these elasticities using a simultaneous-equation model based on aggregate time-series data for Denmark for 1980-2007. Our best estimates of the short run and the long run rebound effects for road freight transportation are 19% and 28%, respectively. We also find that an increase in the fuel price surprisingly has...

  13. Short transmembrane domains with high-volume exoplasmic halves determine retention of Type II membrane proteins in the Golgi complex.

    Science.gov (United States)

    Quiroga, Rodrigo; Trenchi, Alejandra; González Montoro, Ayelén; Valdez Taubas, Javier; Maccioni, Hugo J F

    2013-12-01

    It is still unclear why some proteins that travel along the secretory pathway are retained in the Golgi complex whereas others make their way to the plasma membrane. Recent bioinformatic analyses on a large number of single-spanning membrane proteins support the hypothesis that specific features of the transmembrane domain (TMD) are relevant to the sorting of these proteins to particular organelles. Here we experimentally test this hypothesis for Golgi and plasma membrane proteins. Using the Golgi SNARE protein Sft1 and the plasma membrane SNARE protein Sso1 from Saccharomyces cerevisiae as model proteins, we modified the length of their TMDs and the volume of their exoplasmic hemi-TMD, and determined their subcellular localization both in yeast and mammalian cells. We found that short TMDs with high-volume exoplasmic hemi-TMDs confer Golgi membrane residence, whereas TMDs with low-volume exoplasmic hemi-TMDs, either short or long, confer plasma membrane residence to these proteins. Results indicate that the shape of the exoplasmic hemi-TMD, in addition to the length of the entire TMD, determine retention in the Golgi or exit to the plasma membrane of Type II membrane proteins.

  14. Membrane mechanisms and intracellular signalling in cell volume regulation

    DEFF Research Database (Denmark)

    Hoffmann, Else Kay; Dunham, Philip B.

    1995-01-01

    Volume regulation, Signal transduction, Calcium-calmodulin, Stretch-activated channels, Eicosanoids, Macromolecular crowding, Cytoskeleton, Protein phosphorylation, dephosphorylation.......Volume regulation, Signal transduction, Calcium-calmodulin, Stretch-activated channels, Eicosanoids, Macromolecular crowding, Cytoskeleton, Protein phosphorylation, dephosphorylation....

  15. Evidence for P-Glycoprotein Involvement in Cell Volume Regulation Using Coulter Sizing in Flow Cytometry.

    Science.gov (United States)

    Pasquier, Jennifer; Rioult, Damien; Abu-Kaoud, Nadine; Hoarau-Véchot, Jessica; Marin, Matthieu; Le Foll, Frank

    2015-06-24

    The regulation of cell volume is an essential function that is coupled to a variety of physiological processes such as receptor recycling, excitability and contraction, cell proliferation, migration, and programmed cell death. Under stress, cells undergo emergency swelling and respond to such a phenomenon with a regulatory volume decrease (RVD) where they release cellular ions, and other osmolytes as well as a concomitant loss of water. The link between P-glycoprotein, a transmembrane transporter, and cell volume regulation is controversial, and changes in cells volume are measured using microscopy or electrophysiology. For instance, by using the patch-clamp method, our team demonstrated that chloride currents activated in the RVD were more intense and rapid in a breast cancer cell line overexpressing the P-glycoprotein (P-gp). The Cell Lab Quanta SC is a flow cytometry system that simultaneously measures electronic volume, side scatter and three fluorescent colors; altogether this provides unsurpassed population resolution and accurate cell counting. Therefore, here we propose a novel method to follow cellular volume. By using the Coulter-type channel of the cytometer Cell Lab Quanta SC MPL (multi-platform loading), we demonstrated a role for the P-gp during different osmotic treatments, but also a differential activity of the P-gp through the cell cycle. Altogether, our data strongly suggests a role of P-gp in cell volume regulation.

  16. Evidence for P-Glycoprotein Involvement in Cell Volume Regulation Using Coulter Sizing in Flow Cytometry

    Directory of Open Access Journals (Sweden)

    Jennifer Pasquier

    2015-06-01

    Full Text Available The regulation of cell volume is an essential function that is coupled to a variety of physiological processes such as receptor recycling, excitability and contraction, cell proliferation, migration, and programmed cell death. Under stress, cells undergo emergency swelling and respond to such a phenomenon with a regulatory volume decrease (RVD where they release cellular ions, and other osmolytes as well as a concomitant loss of water. The link between P-glycoprotein, a transmembrane transporter, and cell volume regulation is controversial, and changes in cells volume are measured using microscopy or electrophysiology. For instance, by using the patch-clamp method, our team demonstrated that chloride currents activated in the RVD were more intense and rapid in a breast cancer cell line overexpressing the P-glycoprotein (P-gp. The Cell Lab Quanta SC is a flow cytometry system that simultaneously measures electronic volume, side scatter and three fluorescent colors; altogether this provides unsurpassed population resolution and accurate cell counting. Therefore, here we propose a novel method to follow cellular volume. By using the Coulter-type channel of the cytometer Cell Lab Quanta SC MPL (multi-platform loading, we demonstrated a role for the P-gp during different osmotic treatments, but also a differential activity of the P-gp through the cell cycle. Altogether, our data strongly suggests a role of P-gp in cell volume regulation.

  17. Tumor volume in subcutaneous mouse xenografts measured by microCT is more accurate and reproducible than determined by 18F-FDG-microPET or external caliper

    Directory of Open Access Journals (Sweden)

    Jørgensen Jesper

    2008-10-01

    Full Text Available Abstract Background In animal studies tumor size is used to assess responses to anticancer therapy. Current standard for volumetric measurement of xenografted tumors is by external caliper, a method often affected by error. The aim of the present study was to evaluate if microCT gives more accurate and reproducible measures of tumor size in mice compared with caliper measurements. Furthermore, we evaluated the accuracy of tumor volume determined from 18F-fluorodeoxyglucose (18F-FDG PET. Methods Subcutaneously implanted human breast adenocarcinoma cells in NMRI nude mice served as tumor model. Tumor volume (n = 20 was determined in vivo by external caliper, microCT and 18F-FDG-PET and subsequently reference volume was determined ex vivo. Intra-observer reproducibility of the microCT and caliper methods were determined by acquiring 10 repeated volume measurements. Volumes of a group of tumors (n = 10 were determined independently by two observers to assess inter-observer variation. Results Tumor volume measured by microCT, PET and caliper all correlated with reference volume. No significant bias of microCT measurements compared with the reference was found, whereas both PET and caliper had systematic bias compared to reference volume. Coefficients of variation for intra-observer variation were 7% and 14% for microCT and caliper measurements, respectively. Regression coefficients between observers were 0.97 for microCT and 0.91 for caliper measurements. Conclusion MicroCT was more accurate than both caliper and 18F-FDG-PET for in vivo volumetric measurements of subcutaneous tumors in mice.18F-FDG-PET was considered unsuitable for determination of tumor size. External caliper were inaccurate and encumbered with a significant and size dependent bias. MicroCT was also the most reproducible of the methods.

  18. Determining matrix elements and resonance widths from finite volume: the dangerous mu-terms

    CERN Document Server

    Takacs, G

    2011-01-01

    The standard numerical approach to determining matrix elements of local operators and width of resonances uses the finite volume dependence of energy levels and matrix elements. Finite size corrections that decay exponentially in the volume are usually neglected or taken into account using perturbation expansion in effective field theory. Using two-dimensional sine-Gordon field theory as "toy model" it is shown that some exponential finite size effects could be much larger than previously thought, potentially spoiling the determination of matrix elements in frameworks such as lattice QCD. The particular class of finite size corrections considered here are mu-terms arising from bound state poles in the scattering amplitudes. In sine-Gordon model, these can be explicitly evaluated and shown to explain the observed discrepancies to high precision. It is argued that the effects observed are not special to the two-dimensional setting, but rather depend on general field theoretic features that are common with model...

  19. Determining Physical Properties of the Cell Cortex

    Science.gov (United States)

    Saha, Arnab; Nishikawa, Masatoshi; Behrndt, Martin; Heisenberg, Carl-Philipp; Jülicher, Frank; Grill, Stephan W.

    2016-03-01

    Actin and myosin assemble into a thin layer of a highly dynamic network underneath the membrane of eukaryotic cells. This network generates the forces that drive cell and tissue-scale morphogenetic processes. The effective material properties of this active network determine large-scale deformations and other morphogenetic events. For example,the characteristic time of stress relaxation (the Maxwell time)in the actomyosin sets the time scale of large-scale deformation of the cortex. Similarly, the characteristic length of stress propagation (the hydrodynamic length) sets the length scale of slow deformations, and a large hydrodynamic length is a prerequisite for long-ranged cortical flows. Here we introduce a method to determine physical parameters of the actomyosin cortical layer (in vivo). For this we investigate the relaxation dynamics of the cortex in response to laser ablation in the one-cell-stage {\\it C. elegans} embryo and in the gastrulating zebrafish embryo. These responses can be interpreted using a coarse grained physical description of the cortex in terms of a two dimensional thin film of an active viscoelastic gel. To determine the Maxwell time, the hydrodynamic length and the ratio of active stress and per-area friction, we evaluated the response to laser ablation in two different ways: by quantifying flow and density fields as a function of space and time, and by determining the time evolution of the shape of the ablated region. Importantly, both methods provide best fit physical parameters that are in close agreement with each other and that are similar to previous estimates in the two systems. We provide an accurate and robust means for measuring physical parameters of the actomyosin cortical layer.It can be useful for investigations of actomyosin mechanics at the cellular-scale, but also for providing insights in the active mechanics processes that govern tissue-scale morphogenesis.

  20. A Bacterial Cell Shape-Determining Inhibitor.

    Science.gov (United States)

    Liu, Yanjie; Frirdich, Emilisa; Taylor, Jennifer A; Chan, Anson C K; Blair, Kris M; Vermeulen, Jenny; Ha, Reuben; Murphy, Michael E P; Salama, Nina R; Gaynor, Erin C; Tanner, Martin E

    2016-04-15

    Helicobacter pylori and Campylobacter jejuni are human pathogens and causative agents of gastric ulcers/cancer and gastroenteritis, respectively. Recent studies have uncovered a series of proteases that are responsible for maintaining the helical shape of these organisms. The H. pylori metalloprotease Csd4 and its C. jejuni homologue Pgp1 cleave the amide bond between meso-diaminopimelate and iso-d-glutamic acid in truncated peptidoglycan side chains. Deletion of either csd4 or pgp1 results in bacteria with a straight rod phenotype, a reduced ability to move in viscous media, and reduced pathogenicity. In this work, a phosphinic acid-based pseudodipeptide inhibitor was designed to act as a tetrahedral intermediate analog against the Csd4 enzyme. The phosphinic acid was shown to inhibit the cleavage of the alternate substrate, Ac-l-Ala-iso-d-Glu-meso-Dap, with a Ki value of 1.5 μM. Structural analysis of the Csd4-inhibitor complex shows that the phosphinic acid displaces the zinc-bound water and chelates the metal in a bidentate fashion. The phosphinate oxygens also interact with the key acid/base residue, Glu222, and the oxyanion-stabilizing residue, Arg86. The results are consistent with the "promoted-water pathway" mechanism for carboxypeptidase A catalysis. Studies on cultured bacteria showed that the inhibitor causes significant cell straightening when incubated with H. pylori at millimolar concentrations. A diminished, yet observable, effect on the morphology of C. jejuni was also apparent. Cell straightening was more pronounced with an acapsular C. jejuni mutant strain compared to the wild type, suggesting that the capsule impaired inhibitor accessibility. These studies demonstrate that a highly polar compound is capable of crossing the outer membrane and altering cell shape, presumably by inhibiting cell shape determinant proteases. Peptidoglycan proteases acting as cell shape determinants represent novel targets for the development of antimicrobials

  1. Intracellular Ca2+ responses and cell volume regulation upon cholinergic and purinergic stimulation in an immortalized salivary cell line.

    Science.gov (United States)

    Aure, Marit H; Røed, Asbjørn; Galtung, Hilde Kanli

    2010-06-01

    The water channel aquaporin 5 (AQP5) seems to play a key role in salivary fluid secretion and appears to be critical in the cell volume regulation of acinar cells. Recently, the cation channel transient potential vanilloid receptor 4 (TRPV4) was shown to be functionally connected to AQP5 and also to cell volume regulation in salivary glands. We used the Simian virus 40 (SV40) immortalized cell line SMG C10 from the rat submandibular salivary gland to investigate the effect of ATP and the neurotransmitter analogue carbachol on Ca(2+) signalling and cell volume regulation, as well as the involvement of TRPV4 in the responses. We used fura-2-AM imaging, cell volume measurements, and western blotting. Both carbachol and ATP increased the concentration of intracellular Ca(2+), but no volume changes could be measured. Inhibition of TRPV4 with ruthenium red impaired both ATP- and carbachol-stimulated Ca(2+) signals. Peak Ca(2+) signalling during hyposmotic exposure was significantly decreased following inhibition of TRPV4, while the cells' ability to volume regulate appeared to be unaffected. These results show that in the SMG C10 cells, simulation of nervous stimulation did not induce cell swelling, although the cells had intact volume regulatory mechanisms. Furthermore, even though Ca(2+) signals were not needed for this volume regulation, TRPV4 seems to play a role during ATP and carbachol stimulation.

  2. Morphometry and cell volumes of diatoms from a tropical estuary of India

    Digital Repository Service at National Institute of Oceanography (India)

    RaviKumar, M.S.; Ramaiah, N.; Tang, D.

    Cell volumes and per cell carbon contents of different diatoms (10 centric, 14 pennate and 1 dinoflagellate) collected from Dona Paula Bay in the central west coast of India have been analyzed. Morphometric information on the phytoplankton types...

  3. Optical volume and mass measurements show that mammalian cells swell during mitosis.

    Science.gov (United States)

    Zlotek-Zlotkiewicz, Ewa; Monnier, Sylvain; Cappello, Giovanni; Le Berre, Mael; Piel, Matthieu

    2015-11-23

    The extent, mechanism, and function of cell volume changes during specific cellular events, such as cell migration and cell division, have been poorly studied, mostly because of a lack of adequate techniques. Here we unambiguously report that a large range of mammalian cell types display a significant increase in volume during mitosis (up to 30%). We further show that this increase in volume is tightly linked to the mitotic state of the cell and not to its spread or rounded shape and is independent of the presence of an intact actomyosin cortex. Importantly, this volume increase is not accompanied by an increase in dry mass and thus corresponds to a decrease in cell density. This mitotic swelling might have important consequences for mitotic progression: it might contribute to produce strong pushing forces, allowing mitotic cells to round up; it might also, by lowering cytoplasmic density, contribute to the large change of physicochemical properties observed in mitotic cells.

  4. Analytical Chemistry Laboratory (ACL) procedure compendium. Volume 7, Safety operation procedure for hot cell

    Energy Technology Data Exchange (ETDEWEB)

    1993-08-01

    This volume contains the interim change notice for the safety operation procedure for hot cell. It covers the master-slave manipulators, dry waste removal, cell transfers, hoists, cask handling, liquid waste system, and physical characterization of fluids.

  5. Fuzzy hidden Markov chains segmentation for volume determination and quantitation in PET

    Energy Technology Data Exchange (ETDEWEB)

    Hatt, M [INSERM U650, Laboratoire du Traitement de l' Information Medicale (LaTIM), CHU Morvan, Bat 2bis (I3S), 5 avenue Foch, Brest, 29609 (France); Lamare, F [INSERM U650, Laboratoire du Traitement de l' Information Medicale (LaTIM), CHU Morvan, Bat 2bis (I3S), 5 avenue Foch, Brest, 29609, (France); Boussion, N [INSERM U650, Laboratoire du Traitement de l' Information Medicale (LaTIM), CHU Morvan, Bat 2bis (I3S), 5 avenue Foch, Brest, 29609 (France); Turzo, A [INSERM U650, Laboratoire du Traitement de l' Information Medicale (LaTIM), CHU Morvan, Bat 2bis (I3S), 5 avenue Foch, Brest, 29609 (France); Collet, C [Ecole Nationale Superieure de Physique de Strasbourg (ENSPS), ULP, Strasbourg, F-67000 (France); Salzenstein, F [Institut d' Electronique du Solide et des Systemes (InESS), ULP, Strasbourg, F-67000 (France); Roux, C [INSERM U650, Laboratoire du Traitement de l' Information Medicale (LaTIM), CHU Morvan, Bat 2bis (I3S), 5 avenue Foch, Brest, 29609 (France); Jarritt, P [Medical Physics Agency, Royal Victoria Hospital, Belfast (United Kingdom); Carson, K [Medical Physics Agency, Royal Victoria Hospital, Belfast (United Kingdom); Rest, C Cheze-Le [INSERM U650, Laboratoire du Traitement de l' Information Medicale (LaTIM), CHU Morvan, Bat 2bis (I3S), 5 avenue Foch, Brest, 29609 (France); Visvikis, D [INSERM U650, Laboratoire du Traitement de l' Information Medicale (LaTIM), CHU Morvan, Bat 2bis (I3S), 5 avenue Foch, Brest, 29609 (France)

    2007-07-21

    Accurate volume of interest (VOI) estimation in PET is crucial in different oncology applications such as response to therapy evaluation and radiotherapy treatment planning. The objective of our study was to evaluate the performance of the proposed algorithm for automatic lesion volume delineation; namely the fuzzy hidden Markov chains (FHMC), with that of current state of the art in clinical practice threshold based techniques. As the classical hidden Markov chain (HMC) algorithm, FHMC takes into account noise, voxel intensity and spatial correlation, in order to classify a voxel as background or functional VOI. However the novelty of the fuzzy model consists of the inclusion of an estimation of imprecision, which should subsequently lead to a better modelling of the 'fuzzy' nature of the object of interest boundaries in emission tomography data. The performance of the algorithms has been assessed on both simulated and acquired datasets of the IEC phantom, covering a large range of spherical lesion sizes (from 10 to 37 mm), contrast ratios (4:1 and 8:1) and image noise levels. Both lesion activity recovery and VOI determination tasks were assessed in reconstructed images using two different voxel sizes (8 mm{sup 3} and 64 mm{sup 3}). In order to account for both the functional volume location and its size, the concept of % classification errors was introduced in the evaluation of volume segmentation using the simulated datasets. Results reveal that FHMC performs substantially better than the threshold based methodology for functional volume determination or activity concentration recovery considering a contrast ratio of 4:1 and lesion sizes of <28 mm. Furthermore differences between classification and volume estimation errors evaluated were smaller for the segmented volumes provided by the FHMC algorithm. Finally, the performance of the automatic algorithms was less susceptible to image noise levels in comparison to the threshold based techniques. The

  6. Surface area and volume determination of subgingival calculus using laser fluorescence.

    Science.gov (United States)

    Shakibaie, Fardad; Walsh, Laurence J

    2014-03-01

    Visible red (655 nm) laser fluorescence (LF) devices are currently used for identifying deposits of subgingival calculus on the root surfaces of teeth during dental examination and treatment; however, it is not known how the fluorescence readings produced by commercially available LF systems correlate to the nature of the deposits. This laboratory study explored the correlation between LF digital readings and the surface area and volume of subgingival calculus deposits on teeth. A collection of 30 extracted human posterior teeth with various levels of subgingival deposits of calculus across 240 sites were used in a clinical simulation, with silicone impression material used to replicate periodontal soft tissues. The teeth were scored by two examiners by using three commercial LF systems (DIAGNOdent, DIAGNOdent Pen and KEY3). The silicone was removed, and the teeth were removed for photography at × 20 magnification under white or ultraviolet light. The surface area, thickness, and volume were calculated, and both linear least squares regression and nonlinear (Spearman's rank method) correlation coefficients were determined. Visible red LF digital readings showed better correlation to calculus volume than to surface area. Overall, the best performance was found for the KEY3 system (Spearman coefficient 0.59), compared to the Classic DIAGNOdent (0.56) and the DIAGNOdent Pen (0.49). These results indicate that while visible red LF systems vary somewhat in performance, their LF readings provide a useful estimation of the volume of subgingival calculus deposits present on teeth.

  7. Determination of the Volume of Water for Suppressing the Thermal Decomposition of Forest Combustibles

    Science.gov (United States)

    Volkov, R. S.; Zhdanova, A. O.; Kuznetsov, G. V.; Strizhak, P. A.

    2017-07-01

    From the results of experimental studies of the processes of suppressing the thermal decomposition of the typical forest combustibles (birch leaves, fir needles, asp twigs, and a mixture of these three materials) by water aerosol, the minimum volumes of the fire-extinguishing liquid have been determined (by varying the volume of samples of the forest combustibles from 0.00002 m3 to 0.0003 m3 and the area of their open surface from 0.0001 m2 to 0.018 m2). The dependences of the minimum volume of water on the area of the open surface of the forest combustible have been established. Approximation expressions for these dependences have been obtained. Forecast has been made of the minimum volume of water for suppressing the process of thermal decomposition of forest combustibles in areas from 1 cm2 to 1 km2, as well as of the characteristic quenching times by varying the water concentration per unit time. It has been shown that the amount of water needed for effective suppression of the process of thermal decomposition of forest combustibles is several times less than is customarily assumed.

  8. Modulation of KCNQ4 channel activity by changes in cell volume

    DEFF Research Database (Denmark)

    Hougaard, Charlotte; Klaerke, Dan A; Hoffmann, Else K;

    2004-01-01

    KCNQ4 channels expressed in HEK 293 cells are sensitive to cell volume changes, being activated by swelling and inhibited by shrinkage, respectively. The KCNQ4 channels contribute significantly to the regulatory volume decrease (RVD) process following cell swelling. Under isoosmotic conditions......, the KCNQ4 channel activity is modulated by protein kinases A and C, G protein activation, and a reduction in the intracellular Ca2+ concentration, but these signalling pathways are not responsible for the increased channel activity during cell swelling....

  9. Determination of volume, shape and refractive index of individual blood platelets

    CERN Document Server

    Kolesnikova, Irina V; Yurkin, Maxim A; Hoekstra, Alfons G; Maltsev, Valeri P; Semyanov, Konstantin A

    2008-01-01

    Light scattering patterns (LSP) of blood platelets were theoretically and experimentally analyzed. We used spicular spheroids as a model for the platelets with pseudopodia. The discrete dipole approximation was employed to simulate light scattering from an individual spicular spheroid constructed from a homogeneous oblate spheroid and 14 rectilinear parallelepipeds rising from the cell centre. These parallelepipeds have a weak effect on the LSP over the measured angular range. Therefore, a homogeneous oblate spheroid was taken as a simplified optical model for platelets. Using the T-matrix method, we computed the LSP over a range of volumes, aspect ratios and refractive indices. Measured LSPs of individual platelets were compared one by one with the theoretical set and the best fit was taken to characterize the measured platelets, resulting in distributions of volume, aspect ratio and refractive index.

  10. Tracer techniques for urine volume determination and urine collection and sampling back-up system

    Science.gov (United States)

    Ramirez, R. V.

    1971-01-01

    The feasibility, functionality, and overall accuracy of the use of lithium were investigated as a chemical tracer in urine for providing a means of indirect determination of total urine volume by the atomic absorption spectrophotometry method. Experiments were conducted to investigate the parameters of instrumentation, tracer concentration, mixing times, and methods for incorporating the tracer material in the urine collection bag, and to refine and optimize the urine tracer technique to comply with the Skylab scheme and operational parameters of + or - 2% of volume error and + or - 1% accuracy of amount of tracer added to each container. In addition, a back-up method for urine collection and sampling system was developed and evaluated. This back-up method incorporates the tracer technique for volume determination in event of failure of the primary urine collection and preservation system. One chemical preservative was selected and evaluated as a contingency chemical preservative for the storage of urine in event of failure of the urine cooling system.

  11. Air-kerma determination using a variable-volume cavity ionization chamber standard.

    Science.gov (United States)

    Burns, D T; Kessler, C; Roger, P

    2007-12-01

    A graphite-walled cavity ionization chamber of modular design and variable volume has been used to determine the air-kerma rate in the reference 60Co field at the BIPM. The chamber can be configured in five sizes. High-accuracy mechanical measurements of the volume of the air cavity were made for each configuration using a co-ordinate measuring machine. Ionization current measurements were made for each configuration and corrected for the effects of ion recombination and diffusion, stem scatter and chamber orientation. Monte Carlo calculations of cavity dose were made to evaluate the correction factors kwall and kan. A reproducibility of the ionization current per mass of 1.5 parts in 10(4) was achieved on the repeated assembly of each configuration. The results show an air-kerma rate determination that increases with volume, the total change being around 8 parts in 10(4). When analysed differentially, the air-kerma rate relative to the BIPM standard is Kdiff/KBIPM = 1.0026(6). A detailed uncertainty budget is presented. Possible reasons for the observed behaviour are discussed that might have consequences for all existing standards for air-kerma.

  12. Volume change determination of metastatic lung tumors in CT images using 3-D template matching

    Science.gov (United States)

    Ambrosini, Robert D.; Wang, Peng; O'Dell, Walter G.

    2009-02-01

    The ability of a clinician to properly detect changes in the size of lung nodules over time is a vital element to both the diagnosis of malignant growths and the monitoring of the response of cancerous lesions to therapy. We have developed a novel metastasis sizing algorithm based on 3-D template matching with spherical tumor appearance models that were created to match the expected geometry of the tumors of interest while accounting for potential spatial offsets of nodules in the slice thickness direction. The spherical template that best-fits the overall volume of each lung metastasis was determined through the optimization of the 3-D normalized cross-correlation coefficients (NCCC) calculated between the templates and the nodules. A total of 17 different lung metastases were extracted manually from real patient CT datasets and reconstructed in 3-D using spherical harmonics equations to generate simulated nodules for testing our algorithm. Each metastasis 3-D shape was then subjected to 10%, 25%, 50%, 75% and 90% scaling of its volume to allow for 5 possible volume change combinations relative to the original size per each reconstructed nodule and inserted back into CT datasets with appropriate blurring and noise addition. When plotted against the true volume change, the nodule volume changes calculated by our algorithm for these 85 data points exhibited a high degree of accuracy (slope = 0.9817, R2 = 0.9957). Our results demonstrate that the 3-D template matching method can be an effective, fast, and accurate tool for automated sizing of metastatic tumors.

  13. Remote-Sensing Technique for Determination of the Volume Absorption Coefficient of Turbid Water

    Science.gov (United States)

    Sydor, Michael; Arnone, Robert A.; Gould, Richard W., Jr.; Terrie, Gregory E.; Ladner, Sherwin D.; Wood, Christoper G.

    1998-07-01

    We use remote-sensing reflectance from particulate R rs to determine the volume absorption coefficient a of turbid water in the 400 700-nm spectral region. The calculated and measured values of a ( ) show good agreement for 0 . 5 a 10 (m 1 ). To determine R rs from a particulate, we needed to make corrections for remote-sensing reflectance owing to surface roughness S rs . We determined the average spectral distribution of S rs from the difference in total remote-sensing reflectance measured with and without polarization. The spectral shape of S rs showed an excellent fit to theoretical formulas for glare based on Rayleigh and aerosol scattering from the atmosphere.

  14. Effect of radiologic contrast media on cell volume regulatory mechanisms in human red blood cells.

    Science.gov (United States)

    Galtung, Hilde Kanli; Sørlundsengen, Vibeke; Sakariassen, Kjell S; Benestad, Haakon B

    2002-08-01

    The authors performed this study to evaluate cell volume regulation in human red blood cells (RBCs) after incubation in solutions of three contrast media: iohexol (830 mOsm), ioxaglate (520 mOsm), and iodixanol (300 mOsm). Whole blood sampled from six healthy subjects was exposed to Ringer solutions containing 25% or 5% vol/vol iohexol (final osmolality, 440 or 340 mOsm, respectively), ioxaglate (final osmolality, 395 or 335 mOsm, respectively), iodixanol (final osmolality, 330 or 315 mOsm, respectively), or NaCl (control solutions with the same osmolality as that of the contrast media). In some experiments, control RBCs were subjected to a hyposmotic solution (100 mOsm). RBC volumes were obtained with a Coulter counter. The RBCs showed normal regulatory cell shrinkage after hyposmotically induced swelling. All 25% vol/vol contrast material solutions and their control solutions induced RBC shrinkage (range, 6% +/- 1 [standard error] to 22% +/- 3). The same was true for cells exposed to 5% vol/vol contrast material (range, 4% +/- 1 to 7% +/- 1). The shrinkage phase was followed by cell swelling (10% +/- 2 to 20% +/- 2 for 25% contrast material and their control solutions and 8% +/- 1 to 15% +/- 2 for 5% contrast material and their control solutions). No contrast material-exposed RBCs increased their volumes to the level reached with their control solutions. RBCs exposed to hyperosmotic iohexol, ioxaglate, or iodixanol solutions shrank and then swelled. The degree of shrinkage and subsequent swelling could not be explained simply with the osmolality of the test solutions. Physicochemical properties of the contrast media must be involved, putatively affecting electrolyte fluxes over the RBC membrane. Possible targets of these effects are the K+/Cl- symporter, K+ channels, and the Na+/K+/Cl- symporter.

  15. Effect of vagus nerve stimulation on the secretory-granule volume of the principal cells of the mouse gallbladder epithelium.

    Science.gov (United States)

    Wahlin, T; Hulliger, M; Axelsson, H

    1979-07-01

    Experiments in mice were performed in order to investigate whether vagal activity could affect glycoprotein secretion from gallbladder principal cells. This cell type was studied with the electron microscope in control animals and after electric stimulation of the right or left nervus vagus. The volume density of glycoprotein containing granules was determined using morphometry. It was found that stimulation of the left vagus nerve significantly reduced the relative cellular volume of secretory granules in the principal cells of the gallbladder. Right vagus stimulation was accompanied by a weak but insignificant increase in secretory granule content. It is suggested that the left vagus nerve may exert a direct influence on glycoprotein secretion from gallbladder principal cells.

  16. Lung function, transfusion, pulmonary capillary blood volume and sickle cell disease.

    Science.gov (United States)

    Lunt, Alan; McGhee, Emily; Robinson, Polly; Rees, David; Height, Susan; Greenough, Anne

    2016-02-01

    Lung function abnormalities occur in children with sickle cell disease (SCD) and may be associated with elevated pulmonary blood volume. To investigate that association, we determined whether blood transfusion in SCD children acutely increased pulmonary capillary blood volume (PCBV) and increased respiratory system resistance (Rrs5). Measurements of Rrs5 and spirometry were made before and after blood transfusion in 18 children, median age 14.2 (6.6-18.5) years. Diffusing capacity for carbon monoxide and nitric oxide were assessed to calculate the PCBV. Post transfusion, the median Rrs5 had increased from 127.4 to 141.3% predicted (pvolume from 39.7 to 64.1 ml/m2 (pvolume in one second (p=0.0056) and vital capacity (p=0.0008) decreased. The increase in Rrs5 correlated with the increase in PCBV (r=0.50, p=0.0493). Increased pulmonary capillary blood volume may at least partially explain the lung function abnormalities in SCD children.

  17. Determination of volume fractions in two-phase flows from sound speed measurement

    Energy Technology Data Exchange (ETDEWEB)

    Chaudhuri, Anirban [Los Alamos National Laboratory; Sinha, Dipen N. [Los Alamos National Laboratory; Osterhoudt, Curtis F. [University of Alaska

    2012-08-15

    Accurate measurement of the composition of oil-water emulsions within the process environment is a challenging problem in the oil industry. Ultrasonic techniques are promising because they are non-invasive and can penetrate optically opaque mixtures. This paper presents a method of determining the volume fractions of two immiscible fluids in a homogenized two-phase flow by measuring the speed of sound through the composite fluid along with the instantaneous temperature. Two separate algorithms are developed by representing the composite density as (i) a linear combination of the two densities, and (ii) a non-linear fractional formulation. Both methods lead to a quadratic equation with temperature dependent coefficients, the root of which yields the volume fraction. The densities and sound speeds are calibrated at various temperatures for each fluid component, and the fitted polynomial is used in the final algorithm. We present results when the new algorithm is applied to mixtures of crude oil and process water from two different oil fields, and a comparison of our results with a Coriolis meter; the difference between mean values is less than 1%. Analytical and numerical studies of sensitivity of the calculated volume fraction to temperature changes and calibration errors are also presented.

  18. 2,3-Dihydroxybenzoic acid attenuates kanamycin-induced volume reduction in mouse utricular type I hair cells

    DEFF Research Database (Denmark)

    Severinsen, Stig Åvall; Kirkegaard, Mette; Nyengaard, Jens Randel

    2006-01-01

    injection. Total volume of the utricle, as well as total number of hair and supporting cells, were estimated on light microscopic sections. Total volume and mean volume of hair cell types I and II and supporting cells were estimated on digital transmission electron micrographs. Total volume of the utricular...... macula, hair cell type I and supporting cells decreased significantly in animals injected with kanamycin but not in animals co-treated with DHB. Hair and supporting cell numbers remained unchanged in all three groups. In conclusion, the kanamycin-induced volume reduction of type I hair cells...

  19. Islands of surviving cells within necrotic volume at liver induced by PDT

    Science.gov (United States)

    Ferreira, J.; Kurachi, C.; Zucoloto, S.; Castro e Silva, O., Jr.; Bagnato, V. S.

    2009-06-01

    Tissue heterogeneities as well as distinct metabolic status and cellular types within a tumor may results in a nonhomogenous necrosis. The possibility of PDT surviving cells within a treated volume has relevant clinical significance. The major aim of this study was to analyze, on normal rat liver, the cell viability and surviving after PDT. The porphyrin was injected through the cava vein at concentration of 1.5 mg/Kg. The induced necrosis was qualitatively investigated varying the used drug light interval (30 min, 1, 3, 6, 24 and 36 h) and total delivered dose (20, 50, 100, 150 and 200 J/cm2). A diode laser at 630 nm and irradiance of 150 mW/cm2 was employed. The exposed livers were removed after the animals were killed by anesthesia overdose, 30 h after illumination. Slides were processed by HE analysis for the determination of the overall aspects of the necrotic and non-treated liver. We observed necrosis of central vein and presence of surviving cell around the portal triad within the necrotic volume, suggesting PDT-resistant regions of the tissue. Possible hypothesis for the observation may be: the absence of photosensitizer; insufficient light dose (below threshold); and distinct metabolic status in the portal triad microregions decreases oxygen availability to photodynamic reaction. The cells surrounding portal triad presented a higher resistance even when 200 J/cm2 was applied. In contrast, the cells closer to the central vein after 20 J/cm2 were already susceptible to the action of PDT. Different aspects of the problem are presented.

  20. Effects of methylprednisolone on concanavalin A-induced human lymphocyte blastogenesis: a comparative analysis by flow cytometry, volume determination and /sup 3/H-thymidine incorporation

    Energy Technology Data Exchange (ETDEWEB)

    Marder, P.; Schmidtke, J.R.

    1983-08-01

    The inhibition of concanavalin A-induced human peripheral blood lymphocyte blastogenesis by methylprednisolone (MP) was studied by using flow cytometry and tritiated thymidine (/sup 3/H-TdR) incorporation. Flow cytometric determinations of volume, low angle forward light scatter, and nucleic acid showed MP to be a potent inhibitor of blastogenesis. The effects were concentration-dependent and correlated with /sup 3/H-TdR uptake. By using the single cell analytic capability of flow cytometry, the target stages of the cell cycle where MP affects lymphocyte activation were determined. Evidence is presented that steroids can block both early and late phases of this process.

  1. Determining cardiac fiber orientation using FSL and registered ultrasound/DTI volumes

    Science.gov (United States)

    Dormer, James; Qin, Xulei; Shen, Ming; Wang, Silun; Zhang, Xiaodong; Jiang, Rong; Wagner, Mary B.; Fei, Baowei

    2016-04-01

    Accurate extraction of cardiac fiber orientation from diffusion tensor imaging is important for determining heart structure and function. However, the acquisition of magnetic resonance (MR) diffusion tensor images is costly and time consuming. By comparison, cardiac ultrasound imaging is rapid and relatively inexpensive, but it lacks the capability to directly measure fiber orientations. In order to create a detailed heart model from ultrasound data, a three-dimensional (3D) diffusion tensor imaging (DTI) with known fiber orientations can be registered to an ultrasound volume through a geometric mask. After registration, the cardiac orientations from the template DTI can be mapped to the heart using a deformable transformation field. This process depends heavily on accurate fiber orientation extraction from the DTI. In this study, we use the FMRIB Software Library (FSL) to determine cardiac fiber orientations in diffusion weighted images. For the registration between ultrasound and MRI volumes, we achieved an average Dice similarity coefficient (DSC) of 81.6+/-2.1%. For the estimation of fiber orientations from the proposed method, we achieved an acute angle error (AAE) of 22.7+/-3.1° as compared to the direct measurements from DTI. This work provides a new approach to generate cardiac fiber orientation that may be used for many cardiac applications.

  2. Quantitative absorption cytometry for measuring red blood cell hemoglobin mass and volume.

    Science.gov (United States)

    Schonbrun, Ethan; Malka, Roy; Di Caprio, Giuseppe; Schaak, Diane; Higgins, John M

    2014-04-01

    We present an optical system, called the quantitative absorption cytometer (QAC), to measure the volume and hemoglobin mass of red blood cells flowing through a microfluidic channel. In contrast to clinical hematology analyzers, where cells are sphered in order for both volume and hemoglobin to be measured accurately, the QAC measures cells in their normal physiological shape. Human red blood cells are suspended in a refractive index-matching absorbing buffer, driven through a microfluidic channel, and imaged using a transmission light microscope onto a color camera. A red and a blue LED illuminate cells and images at each color are used to independently retrieve cell volume and hemoglobin mass. This system shows good agreement with red blood cell indices retrieved by a clinical hematology analyzer and in fact measures a smaller coefficient of variation of hemoglobin concentration. In addition to cell indices, the QAC returns height and mass maps of each measured cell. These quantitative images are valuable for analyzing the detailed morphology of individual cells as well as statistical outliers found in the data. We also measured red blood cells in hypertonic and hypotonic buffers to quantify the correlation between volume and hemoglobin mass under osmotic stress. Because this method is invariant to cell shape, even extremely nonspherical cells in hypertonic buffers can be measured accurately.

  3. Role of BK channels in the apoptotic volume decrease in native eel intestinal cells

    DEFF Research Database (Denmark)

    Lionetto, Maria Giulia; Giordano, Maria Elena; Calisi, Antonio

    2010-01-01

    of these channels in the Apoptotic Volume Decrease (AVD) of isolated eel enterocytes, and the possible interaction between BK channels and the progression of apoptosis. The detection of apoptosis was performed by confocal microscopy and annexin V and propidium iodide labelling; cell volume changes were monitored...

  4. Determination of aflatoxin M1 in milk by ultra-performance liquid chromatography and fluorimetric detection combined with large volume flow cell%超高效液相色谱-大体积流通池荧光法检测奶及奶制品中的黄曲霉毒素M1

    Institute of Scientific and Technical Information of China (English)

    王军淋; 蔡增轩; 任一平

    2013-01-01

    affecting the pericellular membrane ,inhibiting the synthesis of RNA and interfering the inductive style of specific enzymes . In December 2011 , the General Administration of Quality Supervision , Inspection and Quarantine of the People s Republic of China ( AQSIQ) announced the selective examination results of 200 kinds of liquid milk products . The aflatoxin M 1 in parts of milk products were over ranging the maximum residue limits ( MRLs) of M 1 in milk and milk products , of which the maximum superscalar were 140% exceeded . Moreover , the contents of aflatoxin M 1 were found exceeded in milk powder again in July 2012 . The method used now was to first heat the milk and milk products in water bath , then samples were clean‐up and concentrated by immunoaffinity column after filtered or centrifuged . In this method , no clear sample solutions were obtained , when passing through the immunoaffinity column , sometimes the immunoaffinity column would be blocked , and the recovery would not in expectation . So a better method was needed for determinating the aflatoxin M 1 in milk and milk products . The present study developed an improved analytical method for the fast determination of aflatoxin M 1 in milk and milk products by ultra‐performance liquid chromatography ( UPLC ) combined with large volume flow cell fluorescence detection ( FLD) . The milk sample was extracted by acetonitrile , and the ratio of the milk sample to acetonitrile was 1 to 2 .5 in mass to volume . Then , the sample was extracted by vortex and ultrasound assisted liquid‐liquid extraction ( ULLE) , and was cleaned‐up and concentrated by aflatoxin M 1 immunoaffinity column . The analyte was separated by UPLC BEH C18 column (100 mm × 2 .1 mm , 1 .7μm) , and was eluted with acetonitrile‐methanol ( 50∶50) and pure water . The results showed that the limit of quantitation ( LOQ) of aflatoxin M 1 was 0 .03 μg/kg , which was lower than the national criteria on determination of the minimum level

  5. Preoperative determination of prostate cancer tumor volume: analysis through biopsy fragments

    Directory of Open Access Journals (Sweden)

    Alberto A. Antunes

    2007-08-01

    Full Text Available OBJECTIVE: Preoperative determination of prostate cancer (PCa tumor volume (TV is still a big challenge. We have assessed variables obtained in prostatic biopsy aiming at determining which is the best method to predict the TV in radical prostatectomy (RP specimens. MATERIALS AND METHODS: Biopsy findings of 162 men with PCa submitted to radical prostatectomy were revised. Preoperative characteristics, such as PSA, the percentage of positive fragments (PPF, the total percentage of cancer in the biopsy (TPC, the maximum percentage of cancer in a fragment (MPC, the presence of perineural invasion (PNI and the Gleason score were correlated with postoperative surgical findings through an univariate analysis of a linear regression model. RESULTS: The TV correlated significantly to the PPF, TPC, MPC, PSA and to the presence of PNI (p < 0.001. However, the Pearson correlation analysis test showed an R2 of only 24%, 12%, 17% and 9% for the PPF, TPC, MPC, and PSA respectively. The combination of the PPF with the PSA and the PNI analysis showed to be a better model to predict the TV (R2 of 32.3%. The TV could be determined through the formula: Volume = 1.108 + 0.203 x PSA + 0.066 x PPF + 2.193 x PNI. CONCLUSIONS: The PPF seems to be better than the TPC and the MPC to predict the TV in the surgical specimen. Due to the weak correlation between those variables and the TV, the PSA and the presence of PNI should be used together.

  6. Modeling Degradation in Solid Oxide Electrolysis Cells - Volume II

    Energy Technology Data Exchange (ETDEWEB)

    Manohar Motwani

    2011-09-01

    Idaho National Laboratory has an ongoing project to generate hydrogen from steam using solid oxide electrolysis cells (SOECs). To accomplish this, technical and degradation issues associated with the SOECs will need to be addressed. This report covers various approaches being pursued to model degradation issues in SOECs. An electrochemical model for degradation of SOECs is presented. The model is based on concepts in local thermodynamic equilibrium in systems otherwise in global thermodynamic non-equilibrium. It is shown that electronic conduction through the electrolyte, however small, must be taken into account for determining local oxygen chemical potential,, within the electrolyte. The within the electrolyte may lie out of bounds in relation to values at the electrodes in the electrolyzer mode. Under certain conditions, high pressures can develop in the electrolyte just near the oxygen electrode/electrolyte interface, leading to oxygen electrode delamination. These predictions are in accordance with the reported literature on the subject. Development of high pressures may be avoided by introducing some electronic conduction in the electrolyte. By combining equilibrium thermodynamics, non-equilibrium (diffusion) modeling, and first-principles, atomic scale calculations were performed to understand the degradation mechanisms and provide practical recommendations on how to inhibit and/or completely mitigate them.

  7. Determining the lymph node clinical target volume of upper esophageal carcinoma with computed tomography

    Directory of Open Access Journals (Sweden)

    Li Minghuan

    2013-01-01

    Full Text Available Radiation is an important modality for cervical and upper-thoracic esophageal squamous cell carcinoma (ESCC. Delineating the lymph node clinical target volume (CTVn for EC remains a challenging task. The present paper retrospectively analyzes the distribution of affected lymph nodes of cervical and upper thoracic ESCC on CT images to provide a reference for determination of CTVn. The cases of untreated cervical or upper-thoracic ESCC patients with regional lymph node metastases at diagnosis were retrospectively analyzed. CT scans were done to assess the extent of circumferential involvement and the local-regional lymph node status. Based on the CT criteria (cervical, mediastinal and upper abdominal lymph nodes were considered to be positive for malignancy when they were larger than 8-12 mm in short-axis diameter according to different station respectively. Detailed lymph node stations were recorded for every case and the distribution information of loco-regional node metastasis for these patients was analyzed. A total of 256 patients were diagnosed with node metastasis and qualified for the study, including 206 men and 50 women (age range 37-85 years, median 60. This included 205 upper thoracic cases and 51 of cervical lesion. The length of the primary tumors ranged from 1.0 cm to 9.0 cm, median 4.5 cm. The size of the enlarged lymph nodes ranged from 0.8 to 5.0 cm median 1.4 cm, mean 1.61 cm. The number of involved stations ranged from 1 to 7 median 2. The lymph node stations, with an involved probability of 10% or more, included the upper and middle neck, supraclavicular and lower neck, upper paraesophageal and upper paratracheal area for cervical lesions, and the supraclavicular and lower neck, upper paraesophageal, upper paratracheal, lower paratracheal, aortopulmonary and subcarinal areas for upper thoracic EC, respectively. The mid-upper neck nodes were more likely to be involved in cervical EC than thoracic EC (X 2 test, p=0.000. Fewer

  8. Regulatory Volume Decrease in Neural Precursor Cells: Taurine Efflux and Gene Microarray Analysis

    Directory of Open Access Journals (Sweden)

    Reyna Hernández-Benítez

    2014-11-01

    Full Text Available Background/Aims: Neural stem/ progenitor cells (NPCs endure important changes in cell volume during growth, proliferation and migration. As a first approach to know about NPC response to cell volume changes, the Regulatory Volume Decrease (RVD subsequent to hypotonic swelling was investigated. Methods: NPCs obtained from the mesencephalon and the subventricular zone of embryonic and adult mice, respectively, were grown and cultured as neurospheres. Cell volume changes were measured by large-angle light-scattering and taurine efflux by [3H]-taurine. Expression of genes encoding molecules related to RVD was analysed using a DNA microarray obtained from NPC samples. Results: Embryonic and adult NPCs exposed to osmolarity reduction (H15, H30, H40 exhibited rapid swelling followed by RVD. The magnitude, efficiency and pharmacological profile, of RVD and of [3H]-taurine osmosensitive efflux were comparable to those found in cultured brain cells, astrocytes and neurons. The relative expression of genes encoding molecules related to volume regulation, i.e. K+ and Cl- channels, cotransporters, exchangers and aquaporins were identified in NPCs. Conclusion: NPCs show the ability to respond to hypotonic-evoked volume changes by adaptative recovery processes, similar to those found in other cultured brain cells. Genes related to molecules involved in RVD were found expressed in NPCs.

  9. Power law relationship between cell cycle duration and cell volume in the early embryonic development of Caenorhabditis elegans

    Directory of Open Access Journals (Sweden)

    Yukinobu eArata

    2015-01-01

    Full Text Available Cell size is a critical factor for cell cycle regulation. In Xenopus embryos after midblastula transition, the cell cycle duration elongates in a power law relationship with the cell radius squared. This correlation has been explained by the model that cell surface area is a candidate to determine cell cycle duration. However, it remains unknown whether this second power law is conserved in other animal embryos. Here, we found that the relationship between cell cycle duration and cell size in Caenorhabditis elegans embryos exhibited a power law distribution. Interestingly, the powers of the time-size relationship could be grouped into at least three classes: highly size-correlated, moderately size-correlated, and potentially a size-noncorrelated class according to C. elegans founder cell lineages (1.2, 0.81, and Xenopus and C. elegans, while the absolute powers in C. elegans were different from that in Xenopus. Furthermore, we found that the volume ratio between the nucleus and cell exhibited a power law relationship in the size-correlated classes. The power of the volume relationship was closest to that of the time-size relationship in the highly size-correlated class. This correlation raised the possibility that the time-size relationship, at least in the highly size-correlated class, is explained by the volume ratio of nuclear size and cell size. Thus, our quantitative measurements shed a light on the possibility that early embryonic C. elegans cell cycle duration is coordinated with cell size as a result of geometric constraints between intracellular structures.

  10. Measurement of Single Cell Refractive Index, Dry Mass, Volume, and Density Using a Transillumination Microscope

    Science.gov (United States)

    Phillips, Kevin G.; Jacques, Steven L.; McCarty, Owen J. T.

    2012-09-01

    Phase contrast microscopy has become ubiquitous in the field of biology, particularly in qualitative investigations of cellular morphology. However, the use of quantitative phase retrieval methods and their connection to cellular refractive index and dry mass density remain under utilized. This is due in part to the restriction of phase and cellular mass determination to custom built instruments, involved mathematical analysis, and prohibitive sample perturbations. We introduce tomographic bright field imaging, an accessible optical imaging technique enabling the three dimensional measurement of cellular refractive index and dry mass density using a standard transillumination optical microscope. The validity of the technique is demonstrated on polystyrene spheres. The technique is then applied to the measurement of the refractive index, dry mass, volume, and density of red blood cells. This optical technique enables a simple and robust means to perform quantitative investigations of engineered and biological specimens in three dimensions using standard optical microscopes.

  11. Determination of 137Cs in large volume seawater using Cu-hexacyanoferrate cartridge filters

    Science.gov (United States)

    Visetpotjanakit, S.; Tumnoi, Y.

    2017-06-01

    A simple method to determine 137Cs in seawater has been developed based on the use of a Cu-hexacyanoferrate scavenger. The Cu-hexacyanoferrate supported on cotton wound cartridge filter was used to absorb 137Cs from seawater by passing large volumes over the cartridge filters with flowrate of 240 L hr-1. Results from the Cu-hexacyanoferrate method were proved acceptable for accuracy with bias below ± 20 % i.e. - 9.16 to + 18.55 % when compared with the traditional ammonium molybdophosphate pre-concentration method. This developed method is cost-effective and less time consuming. In addition it can be easily performed at sampling fields.

  12. Using CO to Determine Inhaled Contaminant Volumes and Blower Effectiveness in Several Types of Respirators

    Directory of Open Access Journals (Sweden)

    Arthur T. Johnson

    2011-01-01

    Full Text Available This experiment was conducted to determine how much contaminant could be expected to be inhaled when overbreathing several different types of respirators. These included several tight-fitting and loose-fitting powered air-purifying respirators (PAPRs and one air-purifying respirator (APR. CO2 was used as a tracer gas in the ambient air, and several loose-and tight-fitting respirators were tested on the head form of a breathing machine. CO2 concentration in the exhaled breath was monitored as well as CO2 concentration in the ambient air. This concentration ratio was able to give a measurement of protection factor, not for the respirator necessarily, but for the wearer. Flow rates in the filter/blower inlet and breathing machine outlet were also monitored, so blower effectiveness (defined as the blower contribution to inhaled air could also be determined. Wearer protection factors were found to range from 1.1 for the Racal AirMate loose-fitting PAPR to infinity for the 3M Hood, 3M Breath-Easy PAPR, and SE 400 breath-responsive PAPR. Inhaled contaminant volumes depended on tidal volume but ranged from 2.02 L to 0 L for the same respirators, respectively. Blower effectiveness was about 1.0 for tight-fitting APRs, 0.18 for the Racal, and greater than 1.0 for two of the loose-fitting PAPRs. With blower effectiveness greater than 1.0, some blower flow during the exhalation phase contributes to the subsequent inhalation. Results from this experiment point to different ways to measure respirator efficacy.

  13. Using CO(2) to determine inhaled contaminant volumes and blower effectiveness in several types of respirators.

    Science.gov (United States)

    Johnson, Arthur T; Koh, Frank C; Scott, William H; Rehak, Timothy E

    2011-01-01

    This experiment was conducted to determine how much contaminant could be expected to be inhaled when overbreathing several different types of respirators. These included several tight-fitting and loose-fitting powered air-purifying respirators (PAPRs) and one air-purifying respirator (APR). CO(2) was used as a tracer gas in the ambient air, and several loose-and tight-fitting respirators were tested on the head form of a breathing machine. CO(2) concentration in the exhaled breath was monitored as well as CO(2) concentration in the ambient air. This concentration ratio was able to give a measurement of protection factor, not for the respirator necessarily, but for the wearer. Flow rates in the filter/blower inlet and breathing machine outlet were also monitored, so blower effectiveness (defined as the blower contribution to inhaled air) could also be determined. Wearer protection factors were found to range from 1.1 for the Racal AirMate loose-fitting PAPR to infinity for the 3M Hood, 3M Breath-Easy PAPR, and SE 400 breath-responsive PAPR. Inhaled contaminant volumes depended on tidal volume but ranged from 2.02  L to 0  L for the same respirators, respectively. Blower effectiveness was about 1.0 for tight-fitting APRs, 0.18 for the Racal, and greater than 1.0 for two of the loose-fitting PAPRs. With blower effectiveness greater than 1.0, some blower flow during the exhalation phase contributes to the subsequent inhalation. Results from this experiment point to different ways to measure respirator efficacy.

  14. Using CO2 to Determine Inhaled Contaminant Volumes and Blower Effectiveness in Several Types of Respirators

    Science.gov (United States)

    Johnson, Arthur T.; Koh, Frank C.; Scott, William H.; Rehak, Timothy E.

    2011-01-01

    This experiment was conducted to determine how much contaminant could be expected to be inhaled when overbreathing several different types of respirators. These included several tight-fitting and loose-fitting powered air-purifying respirators (PAPRs) and one air-purifying respirator (APR). CO2 was used as a tracer gas in the ambient air, and several loose-and tight-fitting respirators were tested on the head form of a breathing machine. CO2 concentration in the exhaled breath was monitored as well as CO2 concentration in the ambient air. This concentration ratio was able to give a measurement of protection factor, not for the respirator necessarily, but for the wearer. Flow rates in the filter/blower inlet and breathing machine outlet were also monitored, so blower effectiveness (defined as the blower contribution to inhaled air) could also be determined. Wearer protection factors were found to range from 1.1 for the Racal AirMate loose-fitting PAPR to infinity for the 3M Hood, 3M Breath-Easy PAPR, and SE 400 breath-responsive PAPR. Inhaled contaminant volumes depended on tidal volume but ranged from 2.02 L to 0 L for the same respirators, respectively. Blower effectiveness was about 1.0 for tight-fitting APRs, 0.18 for the Racal, and greater than 1.0 for two of the loose-fitting PAPRs. With blower effectiveness greater than 1.0, some blower flow during the exhalation phase contributes to the subsequent inhalation. Results from this experiment point to different ways to measure respirator efficacy. PMID:21792358

  15. 30 CFR 254.47 - Determining the volume of oil of your worst case discharge scenario.

    Science.gov (United States)

    2010-07-01

    ... drilling operations, the size of your worst case discharge scenario is the daily volume possible from an... break. You must calculate this volume as follows: (1) Add the pipeline system leak detection time to...

  16. Volume regulation in rat pheochromocytoma cultured cells submitted to hypoosmotic conditions.

    Science.gov (United States)

    Delpire, E; Cornet, M; Gilles, R

    1991-02-01

    The mechanisms at work in cell volume regulation have been studied in PC12 cultured cells. Results show, for the first time to our knowledge, that the volume readjustment process occurring after application of a hypoosmotic saline is sensitive to amiloride, IBMX and forskoline. The process is also inhibited by quinine hydrochloride and trifluoperazine. Volume readjustment is concomtant with a decrease in K+ and Cl- intracellular levels. The decrease in K+ level can be related to an assymetrical change in the fluxes in and out of the ion as shown by flux kinetics studies using Rb86. These results are interpreted considering that the control of the activity of the ion channel pathways associated with volume readjustment in PC12 cells may implicate the Ca(2+)-calmodulin - cAMP system.

  17. Anions Govern Cell Volume: A Case Study of Relative Astrocytic and Neuronal Swelling in Spreading Depolarization.

    Directory of Open Access Journals (Sweden)

    Niklas Hübel

    Full Text Available Cell volume changes are ubiquitous in normal and pathological activity of the brain. Nevertheless, we know little about the dynamics of cell and tissue swelling, and the differential changes in the volumes of neurons and glia during pathological states such as spreading depolarizations (SD under ischemic and non-ischemic conditions, and epileptic seizures. By combining the Hodgkin-Huxley type spiking dynamics, dynamic ion concentrations, and simultaneous neuronal and astroglial volume changes into a comprehensive model, we elucidate why glial cells swell more than neurons in SD and the special case of anoxic depolarization (AD, and explore the relative contributions of the two cell types to tissue swelling. Our results demonstrate that anion channels, particularly Cl-, are intrinsically connected to cell swelling and blocking these currents prevents changes in cell volume. The model is based on a simple and physiologically realistic description. We introduce model extensions that are either derived purely from first physical principles of electroneutrality, osmosis, and conservation of particles, or by a phenomenological combination of these principles and known physiological facts. This work provides insights into numerous studies related to neuronal and glial volume changes in SD that otherwise seem contradictory, and is broadly applicable to swelling in other cell types and conditions.

  18. Kinetic assay shows that increasing red cell volume could be a treatment for sickle cell disease

    Science.gov (United States)

    Li, Quan; Henry, Eric R.; Hofrichter, James; Smith, Jeffrey F.; Cellmer, Troy; Dunkelberger, Emily B.; Metaferia, Belhu B.; Jones-Straehle, Stacy; Boutom, Sarah; Christoph, Garrott W.; Wakefield, Terri H.; Link, Mary E.; Staton, Dwayne; Vass, Erica R.; Miller, Jeffery L.; Hsieh, Matthew M.; Tisdale, John F.; Eaton, William A.

    2017-01-01

    Although it has been known for more than 60 years that the cause of sickle cell disease is polymerization of a hemoglobin mutant, hydroxyurea is the only drug approved for treatment by the US Food and Drug Administration. This drug, however, is only partially successful, and the discovery of additional drugs that inhibit fiber formation has been hampered by the lack of a sensitive and quantitative cellular assay. Here, we describe such a method in a 96-well plate format that is based on laser-induced polymerization in sickle trait cells and robust, automated image analysis to detect the precise time at which fibers distort (“sickle”) the cells. With this kinetic method, we show that small increases in cell volume to reduce the hemoglobin concentration can result in therapeutic increases in the delay time prior to fiber formation. We also show that, of the two drugs (AES103 and GBT440) in clinical trials that inhibit polymerization by increasing oxygen affinity, one of them (GBT440) also inhibits sickling in the absence of oxygen by two additional mechanisms. PMID:28096387

  19. Sex determination in mammalian germ cells

    Directory of Open Access Journals (Sweden)

    Cassy M Spiller

    2015-06-01

    Full Text Available Germ cells are the precursors of the sperm and oocytes and hence are critical for survival of the species. In mammals, they are specified during fetal life, migrate to the developing gonads and then undergo a critical period during which they are instructed, by the soma, to adopt the appropriate sexual fate. In a fetal ovary, germ cells enter meiosis and commit to oogenesis, whereas in a fetal testis, they avoid entry into meiosis and instead undergo mitotic arrest and mature toward spermatogenesis. Here, we discuss what we know so far about the regulation of sex-specific differentiation of germ cells, considering extrinsic molecular cues produced by somatic cells, as well as critical intrinsic changes within the germ cells. This review focuses almost exclusively on our understanding of these events in the mouse model.

  20. Determining cell division symmetry through the dissection of dividing cells using single-cell expression analysis.

    Science.gov (United States)

    Jasnos, Lukasz; Sawado, Tomoyuki

    2014-03-01

    Symmetric cell divisions give rise to two sister cells that are identical to each other, whereas asymmetric divisions produce two sister cells with distinctive phenotypes. Although cell division symmetry is usually determined on the basis of a few markers or biological functions, the overall similarity between sister cells has not been thoroughly examined at a molecular level. Here we provide a protocol to separate sister embryonic stem cells (ESCs) and to conduct multiplexed gene expression analyses at the single-cell level by using 48 ESC genes. The procedure includes the dissection of dividing, paired sister cells by micromanipulation, followed by cell lysis, reverse transcription, gene-specific cDNA amplification and multiplexed quantitative PCR analyses. This protocol can be completed in 10 d, and it can be readily adapted to other cell types that are able to grow in suspension culture.

  1. Quantitative analysis of rat adipose tissue cell recovery, and non-fat cell volume, in primary cell cultures

    Directory of Open Access Journals (Sweden)

    Floriana Rotondo

    2016-11-01

    Full Text Available Background White adipose tissue (WAT is a complex, diffuse, multifunctional organ which contains adipocytes, and a large proportion of fat, but also other cell types, active in defense, regeneration and signalling functions. Studies with adipocytes often require their isolation from WAT by breaking up the matrix of collagen fibres; however, it is unclear to what extent adipocyte number in primary cultures correlates with their number in intact WAT, since recovery and viability are often unknown. Experimental Design Epididymal WAT of four young adult rats was used to isolate adipocytes with collagenase. Careful recording of lipid content of tissue, and all fraction volumes and weights, allowed us to trace the amount of initial WAT fat remaining in the cell preparation. Functionality was estimated by incubation with glucose and measurement of glucose uptake and lactate, glycerol and NEFA excretion rates up to 48 h. Non-adipocyte cells were also recovered and their sizes (and those of adipocytes were measured. The presence of non-nucleated cells (erythrocytes was also estimated. Results Cell numbers and sizes were correlated from all fractions to intact WAT. Tracing the lipid content, the recovery of adipocytes in the final, metabolically active, preparation was in the range of 70–75%. Cells showed even higher metabolic activity in the second than in the first day of incubation. Adipocytes were 7%, erythrocytes 66% and other stromal (nucleated cells 27% of total WAT cells. However, their overall volumes were 90%, 0.05%, and 0.2% of WAT. Non-fat volume of adipocytes was 1.3% of WAT. Conclusions The methodology presented here allows for a direct quantitative reference to the original tissue of studies using isolated cells. We have also found that the “live cell mass” of adipose tissue is very small: about 13 µL/g for adipocytes and 2 µL/g stromal, plus about 1 µL/g blood (the rats were killed by exsanguination. These data translate (with

  2. Determination of the efficiency of a detector in gamma spectrometry of large-volume samples

    CERN Document Server

    Tertyshnik, E G

    2012-01-01

    The experimental - calculated method is proposed to determine the full energy peak efficiency (FEPE) of detectors {\\epsilon}(E) in case a measurement of the large-volume samples. Water is used as standard absorber in which the linear attenuation coefficients for photons {\\mu}0 (E) is well known. The value {\\mu} (E) in sample material (matrix of the sample) is determined experimentally by means of spectrometer. The formulas are given for calculation of the ratio {\\epsilon}(E)/ {\\epsilon}0(E), where {\\epsilon}0(E) is FEPE of the detector for photons those are arising in the container filled with water (it is found by adding in the container of the Reference radioactive solutions). To prove the validity of the method ethanol (density 0,8 g/cm3) and water solutions of salts (density 1,2 and 1,5 g/cm3) were used for simulation of the samples with different attenuation coefficients. Standard deviation between experimental and calculated efficiencies has been about 5 %.

  3. Sex determination in mammalian germ cells

    OpenAIRE

    Spiller, Cassy M; Josephine Bowles

    2015-01-01

    Germ cells are the precursors of the sperm and oocytes and hence are critical for survival of the species. In mammals, they are specified during fetal life, migrate to the developing gonads and then undergo a critical period during which they are instructed, by the soma, to adopt the appropriate sexual fate. In a fetal ovary, germ cells enter meiosis and commit to oogenesis, whereas in a fetal testis, they avoid entry into meiosis and instead undergo mitotic arrest and mature toward spermatog...

  4. Determinants of leader cells in collective cell migration.

    NARCIS (Netherlands)

    Khalil, A.; Friedl, P.H.A.

    2010-01-01

    Collective migration is a basic mechanism of cell translocation during morphogenesis, wound repair and cancer invasion. Collective movement requires cells to retain cell-cell contacts, exhibit group polarization with defined front-rear asymmetry, and consequently move as one multicellular unit. Depe

  5. Insensitivity of volume-sensitive chloride currents to chromones in human airway epithelial cells

    Science.gov (United States)

    Zegarra-Moran, Olga; Lantero, Sabina; Sacco, Oliviero; Rossi, Giovanni A; Galietta, Luis J V

    1998-01-01

    Chromones (sodium cromoglycate and sodium nedocromil) block cell swelling-activated Cl− channels in NIH-3T3 fibroblasts and endothelial cells. This has led to hypothesize that cell volume regulation might be involved in asthma pathogenesis.Using whole-cell patch-clamp experiments, we studied the effect of chromones on volume-sensitive Cl− currents in transformed human tracheal epithelial cells (9HTEo-) and in primary cultures of human bronchial epithelial cells (BE).Cl− currents activated by hypotonic shock were poorly blocked by extracellular nedocromil or cromoglycate. The block was voltage-dependent since it was observed only at positive membrane potentials. At the concentration of 5 mM, the current inhibition by both chromones at +80 mV was about 40% for 9HTEo- and only 20% for BE.Intracellular application of chromones elicited a voltage-independent inhibition in 9HTEo- cells. Under this condition, volume-sensitive Cl− currents were reduced at all membrane potentials (60 and 45% inhibition by 2 mM nedocromil and cromoglycate respectively). In contrast intracellular chromones were ineffective in BE cells.The relative refractoriness to chromones, in contrast with the high sensitivity shown by other Cl− channels, suggests that the epithelial volume-sensitive Cl− channel is not involved in asthma. PMID:9863671

  6. The volume of Purkinje cells decreases in the cerebellum of acrylamide-intoxicated rats, but no cells are lost

    DEFF Research Database (Denmark)

    Larsen, Jytte Overgaard; Tandrup, T; Braendgaard, H

    1994-01-01

    The effects of acrylamide intoxication on the numbers of granule and Purkinje cells and the volume of Purkinje cell perikarya have been evaluated with stereological methods. The analysis was carried out in the cerebella of rats that had received a dose of 33.3 mg/kg acrylamide, twice a week, for 7...

  7. Mean reticulocyte volume enhances the utility of red cell mean sphered cell volume in differentiating peripheral blood spherocytes of hereditary spherocytosis from other causes

    Directory of Open Access Journals (Sweden)

    Sukesh C Nair

    2015-01-01

    Full Text Available Context: Mean sphered cell volume (MSCV and mean reticulocyte volume (MRV are additional reticulocyte parameters generated while processing the blood samples on Beckman coulter LH 755 in the reticulocyte mode using the volume, conductivity and scatter technology. It has been observed that the difference between mean corpuscular volume (MCV and MSCV is higher in the cases of hereditary spherocytosis (HS and this difference is increasingly being utilized as a screening tool for spherocytes. In addition now there have been new observations that reticulocyte volume in cases of HS is less as compared to normal reticulocyte. Aims: Our aim was to test the usefulness of reticulocyte parameters like MSCV and MRV in distinguishing cases of HS and autoimmune hemolytic anemia (AIHA. Materials and Methods: This is a retrospective and partly prospective study where peripheral blood ethylenediaminetetraacetic acid samples from cases of HS (n = 57 and AIHA (n = 29 were processed on LH 755 in both the differential and the reticulocyte mode. The data generated were analyzed and compared with data from normal healthy donors (n = 46. Results: Using an algorithm of MCV - MSCV >10 and MRV - MSCV <25, a sensitivity of 84.2% and specificity of 94.7% was observed in cases of HS. Conclusions: With the reticulocyte analysis, we may now have a simple and cheap additional tool for screening of HS.

  8. Determinants of Indices of Cerebral Volume in Former Very Premature Infants at Term Equivalent Age

    Science.gov (United States)

    Wirth, Maelle

    2017-01-01

    Conventional magnetic resonance imaging (MRI) at term equivalent age (TEA) is suggested to be a reliable tool to predict the outcome of very premature infants. The objective of this study was to determine simple reproducible MRI indices, in premature infants and to analyze their neonatal determinants at TEA. A cohort of infants born before 32 weeks gestational age (GA) underwent a MRI at TEA in our center. Two axial images (T2 weighted), were chosen to realize nine measures. We defined 4 linear indices (MAfhlv: thickness of lateral ventricle; CSI: cortex-skull index; VCI: ventricular-cortex index; BOI: bi occipital index) and 1 surface index (VS.A: volume slice area). Perinatal data were recorded. Sixty-nine infants had a GA (median (interquartile range)) of 30.0 weeks GA (27.0; 30.0) and a birth weight of 1240 grams (986; 1477). MRI was done at 41.0 (40.0; 42.0) weeks post menstrual age (PMA). The inter-investigator reproducibility was good. Twenty one MRI (30.5%) were quoted abnormal. We observed an association with retinopathy of prematurity (OR [95CI] = 4.205 [1.231–14.368]; p = 0.017), surgery for patent ductus arteriosus (OR = 4.688 [1.01–21.89]; p = 0.036), early onset infection (OR = 4.688 [1.004–21.889]; p = 0.036) and neonatal treatment by cefotaxime (OR = 3.222 [1.093–9.497]; p = 0.03). There was a difference for VCI between normal and abnormal MRI (0.412 (0.388; 0.429) vs. 0.432 (0.418; 0.449); p = 0,019); BOI was higher when fossa posterior lesions were observed; VS.A seems to be the best surrogate for cerebral volume, 80% of VS.As’ variance being explained by a multiple linear regression model including 7 variables (head circumference at birth and at TEA, PMA, dopamine, ibuprofen treatment, blood and platelets transfusions). These indices, easily and rapidly achievable, seem to be useful but need to be validated in a large population to allow generalization for diagnosis and follow-up of former premature infants. PMID:28125676

  9. Purinergic signalling - a possible mechanism for KCNQ1 channel response to cell volume challenges

    DEFF Research Database (Denmark)

    Bomholtz, Sofia Hammami; Willumsen, Niels J.; Meinild, A.-K.

    2013-01-01

    to ion channel stimulation and cell volume back-regulation. Our aim was to investigate whether volume sensitivity of the voltage-gated K(+) channel, KCNQ1, is dependent on ATP release and regulation by purinergic signalling. METHODS: We used Xenopus oocytes heterologously expressing human KCNQ1, KCNE1......, water channels (AQP1) and P2Y2 receptors. ATP release was monitored by a luciferin-luciferase assay and ion channel conductance was recorded by two-electrode voltage clamp. RESULTS: The luminescence assay showed that oocytes released ATP in response to mechanical, hypoosmotic stimuli and hyperosmotic...... to mechanical stimuli and cell volume changes. Purinergic P2 and P1 receptors confer some of the KCNQ1 channel volume sensitivity, although endogenous adenosine receptors and expressed P2Y2 receptors do so in the negative direction....

  10. Fluorescence exclusion: A simple versatile technique to calculate cell volumes and local heights (Conference Presentation)

    Science.gov (United States)

    Thouvenin, Olivier; Fink, Mathias; Boccara, A. Claude

    2017-02-01

    Understanding volume regulation during mitosis is technically challenging. Indeed, a very sensitive non invasive imaging over time scales ranging from seconds to hours and over large fields is required. Therefore, Quantitative Phase Imaging (QPI) would be a perfect tool for such a project. However, because of asymmetric protein segregation during mitosis, an efficient separation of the refractive index and the height in the phase signal is required. Even though many strategies to make such a separation have been developed, they usually are difficult to implement, have poor sensitivity, or cannot be performed in living cells, or in a single shot. In this paper, we will discuss the use of a new technique called fluorescence exclusion to perform volume measurements. By coupling such technique with a simultaneous phase measurement, we were also able to recover the refractive index inside the cells. Fluorescence exclusion is a versatile and powerful technique that allows the volume measurement of many types of cells. A fluorescent dye, which cannot penetrate inside the cells, is mixed with the external medium in a confined environment. Therefore, the fluorescent signal depends on the inverse of the object's height. We could demonstrate both experimentally and theoretically that fluorescence exclusion can accurately measure cell volumes, even for cells much higher than the depth of focus of the objective. A local accurate height and RI measurement can also be obtained for smaller cells. We will also discuss the way to optimize the confinement of the observation chamber, either mechanically or optically.

  11. Calcium signaling and cell volume regulation are altered in Sjögren's Syndrome.

    Science.gov (United States)

    Enger, Tone Berge; Aure, Marit Høyberg; Jensen, Janicke Liaaen; Galtung, Hilde Kanli

    2014-10-01

    Sjögren's Syndrome (SS) is a chronic autoimmune disease, leading to deficient secretion from salivary and lacrimal glands. Saliva production is normally increased by cholinergic innervation, giving rise to intracellular calcium signaling and water transport through water channels (aquaporins, AQPs). The aim of this study was to investigate possible pathophysiological changes in cell volume regulation, AQP expression and localization, and intracellular calcium signaling in glandular cells from SS patients compared to controls. A total of 35 SS patients and 41 non-SS controls were included. Real time qPCR was combined with immunohistochemistry to analyze the mRNA expression and cellular distribution of AQP1, 3 and 5. Cell volume regulation and intracellular calcium signaling were examined in fresh acinar cells. We show for the first time a reduced mRNA expression of AQP1 and 5 in SS compared to controls, accompanied by a decrease in staining intensity of AQP1, 3 and 5 in areas adjacent to local lymphocytic infiltration. Furthermore, we observed that the SS cells' capacity for volume regulation was abnormal. Similarly, the calcium response after parasympathetic agonist (carbachol) stimulation was markedly decreased in SS cells. It is concluded that mRNA expression of AQP1 and 5, protein distribution of AQP1, 3 and 5, glandular cell volume regulation and intracellular calcium signaling are all altered in SS, pointing to possible pathophysiological mechanisms in SS.

  12. Changes in subcutaneous fat cell volume and insulin sensitivity after weight loss.

    Science.gov (United States)

    Andersson, Daniel P; Eriksson Hogling, Daniel; Thorell, Anders; Toft, Eva; Qvisth, Veronica; Näslund, Erik; Thörne, Anders; Wirén, Mikael; Löfgren, Patrik; Hoffstedt, Johan; Dahlman, Ingrid; Mejhert, Niklas; Rydén, Mikael; Arner, Erik; Arner, Peter

    2014-07-01

    Large subcutaneous fat cells associate with insulin resistance and high risk of developing type 2 diabetes. We investigated if changes in fat cell volume and fat mass correlate with improvements in the metabolic risk profile after bariatric surgery in obese patients. Fat cell volume and number were measured in abdominal subcutaneous adipose tissue in 62 obese women before and 2 years after Roux-en-Y gastric bypass (RYGB). Regional body fat mass by dual-energy X-ray absorptiometry; insulin sensitivity by hyperinsulinemic-euglycemic clamp; and plasma glucose, insulin, and lipid profile were assessed. RYGB decreased body weight by 33%, which was accompanied by decreased adipocyte volume but not number. Fat mass in the measured regions decreased and all metabolic parameters were improved after RYGB (P fat cell size correlated strongly with improved insulin sensitivity (P = 0.0057), regional changes in fat mass did not, except for a weak correlation between changes in visceral fat mass and insulin sensitivity and triglycerides. The curve-linear relationship between fat cell size and fat mass was altered after weight loss (P = 0.03). After bariatric surgery in obese women, a reduction in subcutaneous fat cell volume associates more strongly with improvement of insulin sensitivity than fat mass reduction per se. An altered relationship between adipocyte size and fat mass may be important for improving insulin sensitivity after weight loss. Fat cell size reduction could constitute a target to improve insulin sensitivity. © 2014 by the American Diabetes Association.

  13. Volume measurements of 28Si-enriched spheres using an improved optical interferometer for the determination of the Avogadro constant

    Science.gov (United States)

    Kuramoto, Naoki; Azuma, Yasushi; Inaba, Hajime; Fujii, Kenichi

    2017-04-01

    For the determination of the Avogadro constant by the x-ray crystal density method, an accurate volume measurement of 1 kg Si spheres is of primary importance. For this purpose, an optical interferometer was improved and used to measure the volumes of two 1 kg silicon spheres which were manufactured from a silicon crystal highly enriched in 28Si. The apparent volumes of the spheres, which do not take into account the influence of the surface layers on the volume measurement by interferometry, were determined with a relative standard uncertainty of as small as 2.0  ×  10‑8. The surface of the spheres was characterized by using an improved spectroscopic ellipsometer. By considering the influence of the surface layers, the core volumes of the spheres, which exclude the surface layers, were determined. These results were used for the determination of the Avogadro constant in 2015 as a framework organized by the International Avogadro Coordination project. This paper provides details on the measurements, the improvements made to the apparatus, the data analysis and the uncertainty evaluation.

  14. Determination of protein and solvent volumes in protein crystals from contrast variation data

    Energy Technology Data Exchange (ETDEWEB)

    Badger, J. [Brandeis Univ., Waltham, MA (United States)

    1994-12-31

    By varying the relative values of protein and solvent scattering densities in a crystal, it is possible to obtain information on the shape and dimensions of protein molecular envelopes. Neutron diffraction methods are ideally suited to these contrast variation experiments because H/D exchange leads to large differential changes in the protein and solvent scattering densities and is structurally non-perturbing. Low resolution structure factors have been measured from cubic insulin crystals with differing H/D contents. Structure factors calculated from a simple binary density model, in which uniform scattering densities represent the protein and solvent volumes in the crystals, were compared with these data. The contrast variation differences in the sets of measured structure factors were found to be accurately fitted by this simple model. Trial applications to two problems in crystal structure determination illustrate how this fact may be exploited. (1) A translation function that employs contrast variation data gave a sharp minimum within 1-9{Angstrom} of the correctly positioned insulin molecule and is relatively insensitive to errors in the atomic model. (2) An ab initio phasing method for the contrast variation data, based on analyzing histograms of the density distributions in trial maps, was found to recover the correct molecular envelope.

  15. Cell Volume Regulation and Signaling in 3T3-L1 Pre-adipocytes and Adipocytes

    DEFF Research Database (Denmark)

    Eduardsen, Kathrine; Larsen, Susanne; Novak, Ivana

    2011-01-01

    Caveolae have been implicated in sensing of cell volume perturbations, yet evidence is still limited and findings contradictory. Here, we investigated the possible role of caveolae in cell volume regulation and volume sensitive signaling in an adipocyte system with high (3T3-L1 adipocytes......) and osmotic cell shrinkage (RVI), accompanied by increased swelling-activated taurine efflux. However, caveolin-1 distribution was not detectably altered after osmotic swelling or shrinkage, and caveolae integrity, as studied by cholesterol depletion or expression of dominant negative Cav-1, was not required...... no detectable effect on InsR, yet increased ERK1/2 phosphorylation. In conclusion, differentiated 3T3-L1 adipocytes exhibit greatly accelerated RVD and RVI responses and increased swelling-activated taurine efflux compared to pre-adipocytes. Furthermore, in pre-adipocytes, Cav-1/caveolae integrity...

  16. Red cell volume with changes in plasma osmolarity during maximal exercise.

    Science.gov (United States)

    Van Beaumont, W.

    1973-01-01

    The volume of the red cell in vivo was measured during acute changes in plasma osmolarity evoked through short (6 to 8 min) maximal exercise in six male volunteer subjects. Simultaneous measurements of mean corpuscular red cell volume (MCV), hematocrit, blood hemoglobin, mean corpuscular hemoglobin concentration (MCHC), and plasma osmolarity showed that there was no change in the MCV or MCHC with a concomitant rise of nearly 6% in plasma osmolarity. Apparently, in vivo, the volume of the red cell in exercising healthy human subjects does not change measurably, in spite of significant changes in osmotic pressure of the surrounding medium. Consequently, it is not justified to correct postexercise hematocrit measurements for changes in plasma osmolarity.

  17. Red cell volume with changes in plasma osmolarity during maximal exercise.

    Science.gov (United States)

    Van Beaumont, W.

    1973-01-01

    The volume of the red cell in vivo was measured during acute changes in plasma osmolarity evoked through short (6 to 8 min) maximal exercise in six male volunteer subjects. Simultaneous measurements of mean corpuscular red cell volume (MCV), hematocrit, blood hemoglobin, mean corpuscular hemoglobin concentration (MCHC), and plasma osmolarity showed that there was no change in the MCV or MCHC with a concomitant rise of nearly 6% in plasma osmolarity. Apparently, in vivo, the volume of the red cell in exercising healthy human subjects does not change measurably, in spite of significant changes in osmotic pressure of the surrounding medium. Consequently, it is not justified to correct postexercise hematocrit measurements for changes in plasma osmolarity.

  18. Cell fate determination in the Caenorhabditis elegans epidermal lineages

    NARCIS (Netherlands)

    Soete, G.A.J.

    2007-01-01

    The starting point for this work was to use the hypodermal seam of C. elegans as a model system to study cell fate determination. Even though the seam is a relatively simple developmental system, the mechanisms that control cell fate determination in the seam lineages are connected in a highly compl

  19. Cell fate determination in the Caenorhabditis elegans epidermal lineages

    NARCIS (Netherlands)

    Soete, G.A.J.

    2007-01-01

    The starting point for this work was to use the hypodermal seam of C. elegans as a model system to study cell fate determination. Even though the seam is a relatively simple developmental system, the mechanisms that control cell fate determination in the seam lineages are connected in a highly compl

  20. In vitro and in vivo accuracy of sonographic gallbladder volume determinations

    DEFF Research Database (Denmark)

    Andersen, I B; Monrad, H; Grønvall, S;

    1993-01-01

    being more precise. The absolute deviation was independent of the size of the volume and of the shape of the gallbladder. In vivo Simpson's method was validated on 11 patients with cholecystitis. The gallbladder volumes (mean 65 mL; Range 20 mL to 130 mL) measured by sonography differed from...

  1. Two-dimensional echocardiographic determination of right atrial emptying volume: a noninvasive index in quantifying the degree of tricuspid regurgitation.

    Science.gov (United States)

    DePace, N L; Ren, J F; Kotler, M N; Mintz, G S; Kimbiris, D; Kalman, P

    1983-09-01

    Contrast echocardiography and inferior vena cava ultrasonography are useful techniques in diagnosing tricuspid regurgitation (TR) but are not helpful in estimating the severity. Using a computerized light-pen method for tracing the right atrial (RA) border during systole and diastole in the apical 4-chamber view, single-plane volume determinations were calculated in 10 normal subjects (Group I), 18 patients with atrial fibrillation (AF) and no TR (Group II), 14 patients with mitral stenosis and mild TR (Group IIIa), and 8 patients with mitral stenosis and severe TR (Group IIIb). TR was quantitated as absent, mild or severe by contrast right ventriculography. The RA end-systolic volume was 36.4 +/- 13.1 ml in Group I patients, 59.1 +/- 16.8 ml in Group II patients, 76.9 +/- 55.4 ml in Group IIIa patients, and 154.6 +/- 57.3 ml in Group IIIb patients (all Groups versus Group I, p less than 0.001). The mean RA emptying volume, which equals RA end-systolic volume--RA end-diastolic volume, was 15.3 +/- 5.0 for Group I, 17.7 +/- 3.0 for Group II, 30.4 +/- 8.0 for Group IIIa, and 71.6 +/- 25.4 for Group IIIb. All 8 patients with severe TR but none of the 14 patients with mild TR had an RA emptying volume greater than 40 ml (p less than 0.001). In addition, all 28 patients in Groups I and II but only 4 of 14 patients in Group III had an RA emptying volume less than 26 ml (p less than 0.01). The mean RA pressure measured at cardiac catheterization correlated with RA emptying volume (r = 0.71, p less than 0.001). Thus, RA emptying volume is useful for separating severe TR from mild TR in patients with mitral stenosis.

  2. Quantifying volume reduction and peak flow mitigation for three bioretention cells in clay soils in northeast Ohio.

    Science.gov (United States)

    Winston, Ryan J; Dorsey, Jay D; Hunt, William F

    2016-05-15

    Green infrastructure aims to restore watershed hydrologic function by more closely mimicking pre-development groundwater recharge and evapotranspiration (ET). Bioretention has become a popular stormwater control due to its ability to reduce runoff volume through these pathways. Three bioretention cells constructed in low permeability soils in northeast Ohio were monitored for non-winter quantification of inflow, drainage, ET, and exfiltration. The inclusion of an internal water storage (IWS) zone allowed the three cells to reduce runoff by 59%, 42%, and 36% over the monitoring period, in spite of the tight underlying soils. The exfiltration rate and the IWS zone thickness were the primary determinants of volume reduction performance. Post-construction measured drawdown rates were higher than pre-construction soil vertical hydraulic conductivity tests in all cases, due to lateral exfiltration from the IWS zones and ET, which are not typically accounted for in pre-construction soil testing. The minimum rainfall depths required to produce outflow for the three cells were 5.5, 7.4, and 13.8mm. During events with 1-year design rainfall intensities, peak flow reduction varied from 24 to 96%, with the best mitigation during events where peak rainfall rate occurred before the centroid of the rainfall volume, when adequate bowl storage was available to limit overflow.

  3. Peripheral Lymphoid Volume Expansion and Maintenance Are Controlled by Gut Microbiota via RALDH+ Dendritic Cells.

    Science.gov (United States)

    Zhang, Zongde; Li, Jianjian; Zheng, Wencheng; Zhao, Guang; Zhang, Hong; Wang, Xiaofei; Guo, Yaqian; Qin, Chuan; Shi, Yan

    2016-02-16

    Lymphocyte homing to draining lymph nodes is critical for the initiation of immune responses. Secondary lymphoid organs of germ-free mice are underdeveloped. How gut commensal microbes remotely regulate cellularity and volume of secondary lymphoid organs remains unknown. We report here that, driven by commensal fungi, a wave of CD45(+)CD103(+)RALDH(+) cells migrates to the peripheral lymph nodes after birth. The arrival of these cells introduces high amounts of retinoic acid, mediates the neonatal to adult addressin switch on endothelial cells, and directs the homing of lymphocytes to both gut-associated lymphoid tissues and peripheral lymph nodes. In adult mice, a small number of these RALDH(+) cells might serve to maintain the volume of secondary lymphoid organs. Homing deficiency of these cells was associated with lymph node attrition in vitamin-A-deficient mice, suggesting a perpetual dependence on retinoic acid signaling for structural and functional maintenance of peripheral immune organs.

  4. Chronic effects of ultraviolet-B radiation on growth and cell volume of Phaeodactylum tricornutum (Bacillariophyceae)

    Energy Technology Data Exchange (ETDEWEB)

    Behrenfeld, M.J. (AScI Corp., Newport, OR (United States)); Hardy, J.T. (Western Washington Univ., Bellingham (United States)); Lee, H. II (Environmental Protection Agency, Newport, OR (United States))

    1992-12-01

    Cultures of the temperate estuarine diatom, Phaeodactylum tricornutum Bohlin (NEPCC Clone 31), were grown under ambient intensities of ultraviolet-A radiation (UVAR), photosynthetically active radiation (PAR), and various intensities of ultraviolet-B radiation (UVBR; 290-320 nm). Growth rates and cell volumes were monitored for 36 d. UVBR decreased growth rates and increased cell volumes. Sensitivity of growth to UVBR increased with time. Growth rates of P. tricornutum decreased with increasing ratios of UVBR:UVAR + PAR. 25 refs., 4 figs., 1 tab.

  5. Induction of Inducible Nitric Oxide Synthase by Lipopolysaccharide and the Influences of Cell Volume Changes, Stress Hormones and Oxidative Stress on Nitric Oxide Efflux from the Perfused Liver of Air-Breathing Catfish, Heteropneustes fossilis: e0150469

    National Research Council Canada - National Science Library

    Mahua G Choudhury; Nirmalendu Saha

    2016-01-01

    ...) (a bacterial endotoxin), and (b) to determine the effects of hepatic cell volume changes due to anisotonicity or by infusion of certain metabolites, stress hormones and by induction of oxidative stress on production of NO from...

  6. Cell type-specific adaptation of cellular and nuclear volume in micro-engineered 3D environments.

    Science.gov (United States)

    Greiner, Alexandra M; Klein, Franziska; Gudzenko, Tetyana; Richter, Benjamin; Striebel, Thomas; Wundari, Bayu G; Autenrieth, Tatjana J; Wegener, Martin; Franz, Clemens M; Bastmeyer, Martin

    2015-11-01

    Bio-functionalized three-dimensional (3D) structures fabricated by direct laser writing (DLW) are structurally and mechanically well-defined and ideal for systematically investigating the influence of three-dimensionality and substrate stiffness on cell behavior. Here, we show that different fibroblast-like and epithelial cell lines maintain normal proliferation rates and form functional cell-matrix contacts in DLW-fabricated 3D scaffolds of different mechanics and geometry. Furthermore, the molecular composition of cell-matrix contacts forming in these 3D micro-environments and under conventional 2D culture conditions is identical, based on the analysis of several marker proteins (paxillin, phospho-paxillin, phospho-focal adhesion kinase, vinculin, β1-integrin). However, fibroblast-like and epithelial cells differ markedly in the way they adapt their total cell and nuclear volumes in 3D environments. While fibroblast-like cell lines display significantly increased cell and nuclear volumes in 3D substrates compared to 2D substrates, epithelial cells retain similar cell and nuclear volumes in 2D and 3D environments. Despite differential cell volume regulation between fibroblasts and epithelial cells in 3D environments, the nucleus-to-cell (N/C) volume ratios remain constant for all cell types and culture conditions. Thus, changes in cell and nuclear volume during the transition from 2D to 3D environments are strongly cell type-dependent, but independent of scaffold stiffness, while cells maintain the N/C ratio regardless of culture conditions.

  7. Using thermal balance model to determine optimal reactor volume and insulation material needed in a laboratory-scale composting reactor.

    Science.gov (United States)

    Wang, Yongjiang; Pang, Li; Liu, Xinyu; Wang, Yuansheng; Zhou, Kexun; Luo, Fei

    2016-04-01

    A comprehensive model of thermal balance and degradation kinetics was developed to determine the optimal reactor volume and insulation material. Biological heat production and five channels of heat loss were considered in the thermal balance model for a representative reactor. Degradation kinetics was developed to make the model applicable to different types of substrates. Simulation of the model showed that the internal energy accumulation of compost was the significant heat loss channel, following by heat loss through reactor wall, and latent heat of water evaporation. Lower proportion of heat loss occurred through the reactor wall when the reactor volume was larger. Insulating materials with low densities and low conductive coefficients were more desirable for building small reactor systems. Model developed could be used to determine the optimal reactor volume and insulation material needed before the fabrication of a lab-scale composting system.

  8. Sonographic Determination of Residual Bladder Volume after Application of Different Cystotomy Closure Techniques in Dogs

    Directory of Open Access Journals (Sweden)

    IU Khan1*, MA Khan2, SG Bokhari2, A Safdar2, M Shoaib1, H Akbar2, S Aslam2, MA Khan2 and A Noor2

    2016-11-01

    Full Text Available Residual urine volume is measured to diagnose various neurogenic and obstructive disorders of the urinary bladder. However, it is hypothesized that cystotomy closure using inverting patterns decreases intraluminal diameter of urinary bladder which consequently reduce residual bladder volume. This study aimed to investigate the ideal suturing style for cystotomy incision closure which would exert the least effect on residual bladder volume. The effect of various suturing styles on residual bladder volume was studied sonographically. Residual Bladder Volume (RBV was calculated by the formula, i.e. RBV=L×W× (DL+DT/2×0.625, where L=longitudinal diameter, W=transverse diameter, DL= depth at longitudinal diameter, DT=depth at transverse diameter. 24 healthy mongrel dogs were selected and randomly divided into four equal groups A, B, C and D (n=6. In groups A, B and C, the cystotomy incision was subsequently closed by two-layered appositional suturing pattern, two-layered inverting pattern and three layers (using a combination of appositional and inverting styles, respectively. Group-D remained as sham-operated Control. The results clearly showed that the three-layered closure technique using a combination of appositional and inverting patterns (Group-C, significantly reduced the bladder volume (P<0.01. Two-layered inverting patterns (Group B, also reduced the bladder volume but not up to a significant level, whereas, the appositional suturing technique (group-A exerted the least effect on residual bladder volume. Conclusively, it was inferred that a two-layered appositional suturing pattern should be preferred for closure of cystotomy incision to avoid significant changes in residual bladder volume.

  9. Liquid chromatography-mass spectrometric determination of rufinamide in low volume plasma samples.

    Science.gov (United States)

    Gáll, Zsolt; Vancea, Szende; Dogaru, Maria T; Szilágyi, Tibor

    2013-12-01

    Quantification of rufinamide in plasma was achieved using a selective and sensitive liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS) method. The chromatographic separation was achieved on a reversed phase column (Zorbax SB-C18 100mm×3mm, 3.5μm) under isocratic conditions. The mobile phase consisted of a mixture of water containing 0.1% formic acid and methanol (50:50, v/v). The mass spectrometric detection of the analyte was in multiple reaction monitoring mode (MRM) using an electrospray positive ionization (ESI positive). The monitored ions were 127m/z derived from 239m/z rufinamide and 108m/z derived from 251m/z the internal standard (lacosamide). Protein precipitation with methanol was applied for sample preparation using only 50μl aliquots. The concentration range was 40-2000ng/ml for rufinamide in plasma. The limit of detection was 1.25ng/ml and the lower limit of quantification was established at 5ng/ml rufinamide concentration. Selectivity and matrix effect was verified using individual human, rat and rabbit plasma samples. Short-term, post-preparative and freeze-thaw stability was also investigated. The proposed method provides accuracy, precision and high-throughput (short runtime 4.5min) for quantitative determination of rufinamide in plasma. This is the first reported liquid chromatography-tandem mass spectrometric (LC-MS/MS) method for analysis of rufinamide from low volume plasma samples. The LC-MS/MS method was validated according to the current official guidelines and can be applied to accurately measure rufinamide level of large number of plasma samples from clinical studies or therapeutic drug monitoring.

  10. Cell Polarity Determinants Establish Asymmetry in MEN Signaling

    OpenAIRE

    Monje-Casas, Fernando; Amon, Angelika

    2009-01-01

    Components of the Mitotic Exit Network (MEN), a signaling pathway that triggers exit from mitosis, localize to the spindle pole body (SPB) that migrates into the daughter cell during anaphase but are largely absent from the SPB that remains in the mother cell. Through the analysis of one of the determinants of this asymmetry, Bfa1, we find that the machinery responsible for establishing cell polarity and cytoplasmic microtubules collaborate to establish MEN asymmetry. In cells defective in th...

  11. Method for Determining Language Objectives and Criteria. Volume II. Methodological Tools: Computer Analysis, Data Collection Instruments.

    Science.gov (United States)

    1979-05-25

    This volume presents (1) Methods for computer and hand analysis of numerical language performance data (includes examples) (2) samples of interview, observation, and survey instruments used in collecting language data. (Author)

  12. Role of AQP2 in activation of calcium entry by hypotonicity: implications in cell volume regulation.

    Science.gov (United States)

    Galizia, L; Flamenco, M P; Rivarola, V; Capurro, C; Ford, P

    2008-03-01

    We previously reported in a rat cortical collecting duct cell line (RCCD(1)) that the presence of aquaporin 2 (AQP2) in the cell membrane is critical for the rapid activation of regulatory volume decrease mechanisms (RVD) (Ford et al. Biol Cell 97: 687-697, 2005). The aim of our present work was to investigate the signaling pathway that links AQP2 to this rapid RVD activation. Since it has been previously described that hypotonic conditions induce intracellular calcium ([Ca(2+)](i)) increases in different cell types, we tested the hypothesis that AQP2 could have a role in activation of calcium entry by hypotonicity and its implication in cell volume regulation. Using a fluorescent probe technique, we studied [Ca(2+)](i) and cell volume changes in response to a hypotonic shock in WT-RCCD(1) (not expressing aquaporins) and in AQP2-RCCD(1) (transfected with AQP2) cells. We found that after a hypotonic shock only AQP2-RCCD(1) cells exhibit a substantial increase in [Ca(2+)](i). This [Ca(2+)](i) increase is strongly dependent on extracellular Ca(2+) and is partially inhibited by thapsigargin (1 muM) indicating that the rise in [Ca(2+)](i) reflects both influx from the extracellular medium and release from intracellular stores. Exposure of AQP2-RCCD(1) cells to 100 muM gadolinium reduced the increase in [Ca(2+)](i) suggesting the involvement of a mechanosensitive calcium channel. Furthermore, exposure of cells to all of the above described conditions impaired rapid RVD. We conclude that the expression of AQP2 in the cell membrane is critical to produce the increase in [Ca(2+)](i) which is necessary to activate RVD in RCCD(1) cells.

  13. Distribution and determinants of choroidal thickness and volume using automated segmentation software in a population-based study.

    Science.gov (United States)

    Gupta, Preeti; Jing, Tian; Marziliano, Pina; Cheung, Carol Y; Baskaran, Mani; Lamoureux, Ecosse L; Wong, Tien Yin; Cheung, Chui Ming Gemmy; Cheng, Ching-Yu

    2015-02-01

    To objectively quantify choroidal thickness and choroidal volume using fully automated choroidal segmentation software applied to images obtained from enhanced depth imaging spectral-domain optical coherence tomography (EDI SD OCT) in a population-based study; and evaluate the ocular and systemic determinants of choroidal thickness and choroidal volume. Prospective cross-sectional study. Participants ranging in age from 45 to 85 years were recruited from the Singapore Malay Eye Study-2 (SiMES-2), a follow-up population-based study. All participants (n = 540) underwent a detailed ophthalmic examination, including EDI SD OCT for measurements of thickness and volume of the choroid. The intrasession repeatability of choroidal thickness at 5 measured horizontal locations and macular choroidal volume using automated choroidal segmentation software was excellent (intraclass correlation coefficient, 0.97-0.99). Choroid was significantly thicker under the fovea (242.28 ± 97.58 μm), followed by 3 mm temporal (207.65 ± 80.98 μm), and was thinnest at 3 mm nasal (142.44 ± 79.19 μm) location. The mean choroidal volume at central macular region (within a circle of 1 mm diameter) was 0.185 ± 0.69 mm(3). Among the range of ocular and systemic factors studied, age, sex, and axial length were the only significant predictors of choroidal thickness and choroidal volume (all P choroidal segmentation software, we provide fast, reliable, and objective measurements of choroidal thickness and volume in a population-based sample. Male sex, younger age, and shorter axial length are the factors independently associated with thicker choroid and larger choroidal volume. These factors should be taken into consideration when interpreting EDI SD OCT-based choroidal thickness measurements in clinics. Copyright © 2015 Elsevier Inc. All rights reserved.

  14. Advances in understanding the pathogenesis of the red cell volume disorders.

    Science.gov (United States)

    Badens, Catherine; Guizouarn, Hélène

    2016-09-01

    Genetic defects of erythrocyte transport proteins cause disorders of red blood cell volume that are characterized by abnormal permeability to the cations Na(+) and K(+) and, consequently, by changes in red cell hydration. Clinically, these disorders are associated with chronic haemolytic anaemia of variable severity and significant co-morbidities, such as iron overload. This review provides an overview of recent insights into the molecular basis of this group of rare anaemias involving cation channels and transporters dysfunction. To date, a total of 5 different membrane proteins have been reported to be responsible for volume homeostasis alteration when mutated, 3 of them leading to overhydrated cells (AE1 [also termed SLC4A1], RHAG and GLUT1 [also termed SCL2A1) and 2 others to dehydrated cells (PIEZO1 and the Gardos Channel). These findings are not only of basic scientific interest, but also of direct clinical significance for improving diagnostic procedures and identify potential approaches for novel therapeutic strategies.

  15. On-chip acidification rate measurements from single cardiac cells confined in sub-nanoliter volumes

    OpenAIRE

    Ges, Igor A.; Dzhura, Igor A.; Baudenbacher, Franz J.

    2008-01-01

    The metabolic activity of cells can be monitored by measuring the pH in the extracellular environment. Microfabrication and microfluidic technologies allow the sensor size and the extracellular volumes to be comparable to single cells. A glass substrate with thin film pH sensitive IrOx electrodes was sealed to a replica-molded polydimethylsiloxane (PDMS) microfluidic network with integrated valves. The device, termed NanoPhysiometer, allows the trapping of single cardiac myocytes and the meas...

  16. Statistical representative elementary volumes of porous media determined using greyscale analysis of 3D tomograms

    Science.gov (United States)

    Bruns, S.; Stipp, S. L. S.; Sørensen, H. O.

    2017-09-01

    Digital rock physics carries the dogmatic concept of having to segment volume images for quantitative analysis but segmentation rejects huge amounts of signal information. Information that is essential for the analysis of difficult and marginally resolved samples, such as materials with very small features, is lost during segmentation. In X-ray nanotomography reconstructions of Hod chalk we observed partial volume voxels with an abundance that limits segmentation based analysis. Therefore, we investigated the suitability of greyscale analysis for establishing statistical representative elementary volumes (sREV) for the important petrophysical parameters of this type of chalk, namely porosity, specific surface area and diffusive tortuosity, by using volume images without segmenting the datasets. Instead, grey level intensities were transformed to a voxel level porosity estimate using a Gaussian mixture model. A simple model assumption was made that allowed formulating a two point correlation function for surface area estimates using Bayes' theory. The same assumption enables random walk simulations in the presence of severe partial volume effects. The established sREVs illustrate that in compacted chalk, these simulations cannot be performed in binary representations without increasing the resolution of the imaging system to a point where the spatial restrictions of the represented sample volume render the precision of the measurement unacceptable. We illustrate this by analyzing the origins of variance in the quantitative analysis of volume images, i.e. resolution dependence and intersample and intrasample variance. Although we cannot make any claims on the accuracy of the approach, eliminating the segmentation step from the analysis enables comparative studies with higher precision and repeatability.

  17. Determination of volume fractions of texture components with standard distributions in Euler space

    Science.gov (United States)

    Cho, Jae-Hyung; Rollett, A. D.; Oh, K. H.

    2004-03-01

    The intensities of texture components are modeled by Gaussian distribution functions in Euler space. The multiplicities depend on the relation between the texture component and the crystal and sample symmetry elements. Higher multiplicities are associated with higher maximum values in the orientation distribution function (ODF). The ODF generated by Gaussian function shows that the S component has a multiplicity of 1, the brass and copper components, 2, and the Goss and cube components, 4 in the cubic crystal and orthorhombic sample symmetry. Typical texture components were modeled using standard distributions in Euler space to calculate a discrete ODF, and their volume fractions were collected and verified against the volume used to generate the ODF. The volume fraction of a texture component that has a standard spherical distribution can be collected using the misorientation approach. The misorientation approach means integrating the volume-weighted intensity that is located within a specified cut-off misorientation angle from the ideal orientation. The volume fraction of a sharply peaked texture component can be collected exactly with a small cut-off value, but textures with broad distributions (large full-width at half-maximum (FWHM)) need a larger cut-off value. Larger cut-off values require Euler space to be partitioned between texture components in order to avoid overlapping regions. The misorientation approach can be used for texture's volume in Euler space in a general manner. Fiber texture is also modeled with Gaussian distribution, and it is produced by rotation of a crystal located at g 0, around a sample axis. The volume of fiber texture in wire drawing or extrusion also can be calculated easily in the unit triangle with the angle distance approach.

  18. Application of Nuclear Volume Measurements to Comprehend the Cell Cycle in Root-Knot Nematode-Induced Giant Cells

    Directory of Open Access Journals (Sweden)

    José Dijair Antonino de Souza Junior

    2017-06-01

    Full Text Available Root-knot nematodes induce galls that contain giant-feeding cells harboring multiple enlarged nuclei within the roots of host plants. It is recognized that the cell cycle plays an essential role in the set-up of a peculiar nuclear organization that seemingly steers nematode feeding site induction and development. Functional studies of a large set of cell cycle genes in transgenic lines of the model host Arabidopsis thaliana have contributed to better understand the role of the cell cycle components and their implication in the establishment of functional galls. Mitotic activity mainly occurs during the initial stages of gall development and is followed by an intense endoreduplication phase imperative to produce giant-feeding cells, essential to form vigorous galls. Transgenic lines overexpressing particular cell cycle genes can provoke severe nuclei phenotype changes mainly at later stages of feeding site development. This can result in chaotic nuclear phenotypes affecting their volume. These aberrant nuclear organizations are hampering gall development and nematode maturation. Herein we report on two nuclear volume assessment methods which provide information on the complex changes occurring in nuclei during giant cell development. Although we observed that the data obtained with AMIRA tend to be more detailed than Volumest (Image J, both approaches proved to be highly versatile, allowing to access 3D morphological changes in nuclei of complex tissues and organs. The protocol presented here is based on standard confocal optical sectioning and 3-D image analysis and can be applied to study any volume and shape of cellular organelles in various complex biological specimens. Our results suggest that an increase in giant cell nuclear volume is not solely linked to increasing ploidy levels, but might result from the accumulation of mitotic defects.

  19. A compactly integrated flow cell with a chemiluminescent FIA system for determining lactate concentration in serum.

    Science.gov (United States)

    Nakamura, H; Murakami, Y; Yokoyama, K; Tamiya, E; Karube, I; Suda, M; Uchiyama, S

    2001-01-15

    We have fabricated an integrated flow cell as a total microanalysis system (microTAS). This flow cell (size, 15 x 20 mm; total inner volume, 12.2 microL) was designed for a rational analyzing system of lactate determination for serum. This cell was made by micromachining techniques and consisted of two hollows of a lactate oxidase (LOD) reactor and a mixing cell, a spiral groove, and three penetrated holes. To form the reactor and capillary, these patterns, etched on a silicon wafer, were attached to a glass plate by the anodic bonding method. A photodiode was put under part of the spiral capillary. The compactly accumulated devices were integrated into a flow injection analysis (FIA) system. In the flow cell, lactate was catalyzed to pyruvate and hydrogen peroxide at the LOD reactor; subsequently, hydrogen peroxide reacted with the luminol-ferricyanic reagent at the mixing cell. The resulting chemiluminescent light was detected by the photodiode. Using the miniaturized flow cell, the sample volume for one measurement was greatly reduced to 0.2 microL. The response to lactate was obtained within 30 s and was linear between 0.5 and 5.0 mM (4.5 and 45 mg/dL) lactate with excellent correlative variances of 3.2% (average of three measurements at 5.0 mM). For practical application, the lactate concentration in control human serum was determined using this system. The results showed a good correlation coefficient (r = 0.979) with the results obtained by the spectrophotometric reference method. No difference in sera (normal or pathological) was found. Consequently, this integrated flow cell shows potential as a clinical device for lactate determination in serum. In this article, the effect of the design on the chemiluminescent FIA system is also described.

  20. Rapid determination of benzene derivatives in water samples by trace volume solvent DLLME prior to GC-FID

    Energy Technology Data Exchange (ETDEWEB)

    Diao, Chun Peng; Wei, Chao Hai; Feng, Chun Hua [South China Univ. of Technology, Guangzhou Higher Education Mega Center (China). College of Environmental Science and Engineering; Guangdong Regular Higher Education Institutions, Guangzhou (China). Key Lab. of Environmental Protection and Eco-Remediation

    2012-05-15

    An inexpensive, simple and environmentally friendly method based on dispersive liquid liquid microextraction (DLLME) for rapid determination of benzene derivatives in water samples was proposed. A significant improvement of DLLME procedure was achieved. Trace volume ethyl acetate (60 {mu}L) was exploited as dispersion solvent instead of common ones such as methanol and acetone, the volume of which was more than 0.5 mL, and the organic solvent required in DLLME was reduced to a great extent. Only 83-{mu}L organic solvent was consumed in the whole analytic process and the preconcentration procedure was less than 10 min. The advantageous approach coupled with gas chromatograph-flame ionization detector was proposed for the rapid determination of benzene, toluene, ethylbenzene and xylene isomers in water samples. Results showed that the proposed approach was an efficient method for rapid determination of benzene derivatives in aqueous samples. (orig.)

  1. Idealized Shale Sorption Isotherm Measurements to Determine Pore Volume, Pore Size Distribution, and Surface Area

    Science.gov (United States)

    Holmes, R.; Wang, B.; Aljama, H.; Rupp, E.; Wilcox, J.

    2014-12-01

    One method for mitigating the impacts of anthropogenic CO2-related climate change is the sequestration of CO2 in depleted gas and oil reservoirs, including shale. The accurate characterization of the heterogeneous material properties of shale, including pore volume, surface area, pore size distributions (PSDs) and composition is needed to understand the interaction of CO2 with shale. Idealized powdered shale sorption isotherms were created by varying incremental amounts of four essential components by weight. The first two components, organic carbon and clay, have been shown to be the most important components for CO2 uptake in shales. Organic carbon was represented by kerogen isolated from a Silurian shale, and clay groups were represented by illite from the Green River shale formation. The rest of the idealized shale was composed of equal parts by weight of SiO2 to represent quartz and CaCO3 to represent carbonate components. Baltic, Eagle Ford, and Barnett shale sorption measurements were used to validate the idealized samples. The idealized and validation shale sorption isotherms were measured volumetrically using low pressure N2 (77K) and CO2 (273K) adsorbates on a Quantachrome Autosorb IQ2. Gravimetric isotherms were also produced for a subset of these samples using CO2 and CH4adsorbates under subsurface temperature and pressure conditions using a Rubotherm magnetic suspension balance. Preliminary analyses were inconclusive in validating the idealized samples. This could be a result of conflicting reports of total organic carbon (TOC) content in each sample, a problem stemming from the heterogeneity of the samples and different techniques used for measuring TOC content. The TOC content of the validation samples (Eagle Ford and Barnett) was measured by Rock-Eval pyrolysis at Weatherford Laboratories, while the TOC content in the Baltic validation samples was determined by LECO TOC. Development of a uniform process for measuring TOC in the validation samples is

  2. Design and construct an optical device to determine relative blood volume in patients undergoing hemodialysis.

    Science.gov (United States)

    Dormanesh, Banafshe; Tofangchiha, Shahnaz; Abouei, Vahid; Sharifian, Hani

    2014-04-01

    Occurrence of hypotension during hemodialysis in nearly 20-30% of patients, shows is the necessity of continuous monitoring the patients' blood pressure during hemodialysis. Since directly and non-invasively continuous blood pressure monitoring, is not easy, finding a parameter related to blood pressure, for indirect monitoring is of great value. Related blood volume (RBV) is one of the parameters, related to blood pressure and have a good potential to reflect the patient's hemodynamic condition. The main objective of this study was to design and construct an optical device to determine the RBV in patients undergoing hemodialysis, during the process. After initial studies in order to select a proper sensor, using the ORCAD software, an analog circuit was designed. The implementation and modification of the circuit was done by the clinical tests, using expired blood. Afterwards, for calculation the RBV, controlling the display, data storage and sending it to the computer, an ATmega16 microcontroller was used. For programing the microcontroller, CodeVision software and then Altium Designer software were used for the circuit compression, in order to design the printed circuit board. Finally, all parts of the analog and digital circuit, AC to DC converter and the LCD were embedded in a box. After finalization of the device and before testing it in a real situation, expired blood was used for final evaluation. The evaluation was done by changing the blood concentration, at the start point by adding water to it. In fact, the device can track the changes in blood concentration and display the RBV. After this evaluation, the device was tested in a clinical situation. The results showed there are no interactions between this device and the other devices used in the dialysis section and it can work properly in order to measure the RBV. Considering the hypotension and its consequences in a patient on hemodialysis, solving this problem seems necessary. One method for

  3. Determination Of Partial Molar Volumes Of Iron Species In Silicate Melts

    Science.gov (United States)

    Dwyer, C. A.; Kress, V. C.

    2009-12-01

    One of the major questions of planetary geology is the composition and conditions of the Earth interior. Magmas are among our most important windows into the interior. Iron, being a major element with multiple valence states, is an important influence on redox state. One way to investigate redox state is via a volume equation of state. Density data are used in computing partial molar volumes, which may then be used to construct a volume equation of state. We have devised an apparatus which permits high-quality density measurements in silicate melts at high temperatures (up to ≈1500 °C) and reduced oxygen fugacity at atmospheric pressure using the Archimedean single-bob method. With this apparatus, we have collected the first high-quality density data of iron-bearing silicate melts which had their iron predominately as ferrous. We measured the density of Columbia River Basalt (approximate temperature range: 1250 °C -- 1450 °C) and an artificial composition in the FeO-FeO1.5-CaO-SiO2 system (≈1460 °C) at the oxygen fugacity corresponding to the Iron-Wüstite buffer. We used these data in conjunction with literature density data gathered at thermodynamic equilibrium to calculate the partial molar volume of the iron species as a function of temperature at standard pressure. We speciated iron according to: 0.4 FeO + 0.6 FeO1.5 ↔ FeO1.3 The partial molar volumes of FeO, FeO1.5, and FeO1.3 were fit well with no compositional dependence, despite the wide range of compositions included in this study. To the extent that melt structure can be inferred from partial molar volumes, this result suggests that there is no compositional dependency to the structure of any of these iron species. Our partial molar volumes of FeO and FeO1.5 are consistent within one standard error of those found by Lange & Carmichael (1987) and Ghiorso & Kress (2004). However, our partial molar volume for FeO1.3 at reference temperature was one-third that of Ghiorso & Kress (2004) (Lange

  4. Cell polarity determinants establish asymmetry in MEN signaling.

    Science.gov (United States)

    Monje-Casas, Fernando; Amon, Angelika

    2009-01-01

    Components of the mitotic exit network (MEN), a signaling pathway that triggers exit from mitosis, localize to the spindle pole body (SPB) that migrates into the daughter cell during anaphase but are largely absent from the SPB that remains in the mother cell. Through the analysis of one of the determinants of this asymmetry, Bfa1, we find that the machinery responsible for establishing cell polarity and cytoplasmic microtubules collaborate to establish MEN asymmetry. In cells defective in the Cdc42 signaling pathway or the formin Bni1, Bfa1 localizes to both SPBs. The quantitative analysis of Bfa1 localization further shows that Bfa1 can associate with both SPBs in a transient and highly dynamic fashion, but the protein is stabilized on the SPB that migrates into the daughter cell during anaphase through microtubule-bud cortex interactions. Our results indicate that mother-daughter cell asymmetry determinants establish MEN signaling asymmetry through microtubule-bud cortex interactions.

  5. Diagnostic value of kidney length and volume in sonography for determining kidney scar among infants with urinary infection

    Directory of Open Access Journals (Sweden)

    Mahmoud Jalili

    2011-11-01

    Full Text Available Background: Diagnosis of kidney scar could prevent vesico urethral reflax and its complications such as hypertension and renal failure. This study was conducted to determine average kidney length and volume in infants by sonography to accurate diagnosis of scars.Methods: This study was conducted on infants with the history of UTI who referred to Imam Reza Hospital, Kermanshah Iran, for DMSA scan. They underwent kidney sonography before DMSA scan. Difference in kidney length and volume were calculated and data was recorded and were analyzed using statistical tests.Results: Right kidney length was less than left in 60% of right kidney scars this rate was 77.8% in nonscar patients (P=0.657. Left kidney length was less than right in 50% of left kidney scar and that was 22% in nonscar patients (P=0.241. Kidney volume was smaller than other side in 55.6% of right kidney scar and 62.5% of left kidney scar. Higher length detected in 100% and 47% of right and left kidney inflammation respectively. Higher volume detected in 40% and 76.5% of right and left kidney inflammation.Conclusion: In comparison with other studies suggesting kidney length as a valuble predictor for diagnosis of scar we did not find any relationship between scar and kidney length or volume.

  6. Cell fate determination in zebrafish embryonic and adult muscle development

    NARCIS (Netherlands)

    Tee, J.M.

    2010-01-01

    We are interested in how the genetic basis of muscle precursor cells determines the outcome of the muscle cell fate, and thus leading to disruption in muscle formation and maintenance. We utilized the zebrafish carrying mutations in both Axin1 and Apc1, resulting in overactivation of the Wnt/beta-ca

  7. Minimal volume regulation after shrinkage of red blood cells from five species of reptiles

    DEFF Research Database (Denmark)

    Kristensen, Karina; Berenbrink, Michael; Koldkjær, Pia

    2008-01-01

    Red blood cells (RBCs) from most vertebrates restore volume upon hypertonic shrinkage and the mechanisms underlying this regulatory volume increase (RVI) have been studied extensively in these cells. Despite the phylogenetically interesting position of reptiles, very little is known about their red...... cell function. The present study demonstrates that oxygenated RBCs in all major groups of reptiles exhibit no or a very reduced RVI upon ~ 25% calculated hyperosmotic shrinkage. Thus, RBCs from the snakes Crotalus durissus and Python regius, the turtle Trachemys scripta and the alligator Alligator...... was not characterized. It seems, therefore, that the RVI response based on NHE activation was lost among the early sauropsids that gave rise to modern reptiles and birds, while it was retained in mammals. An RVI response has then reappeared in birds, but based on activation of the NKCC. Alternatively, the absence...

  8. Resonant microchannel volume and mass measurements show that suspended cells swell during mitosis.

    Science.gov (United States)

    Son, Sungmin; Kang, Joon Ho; Oh, Seungeun; Kirschner, Marc W; Mitchison, T J; Manalis, Scott

    2015-11-23

    Osmotic regulation of intracellular water during mitosis is poorly understood because methods for monitoring relevant cellular physical properties with sufficient precision have been limited. Here we use a suspended microchannel resonator to monitor the volume and density of single cells in suspension with a precision of 1% and 0.03%, respectively. We find that for transformed murine lymphocytic leukemia and mouse pro-B cell lymphoid cell lines, mitotic cells reversibly increase their volume by more than 10% and decrease their density by 0.4% over a 20-min period. This response is correlated with the mitotic cell cycle but is not coupled to nuclear osmolytes released by nuclear envelope breakdown, chromatin condensation, or cytokinesis and does not result from endocytosis of the surrounding fluid. Inhibiting Na-H exchange eliminates the response. Although mitotic rounding of adherent cells is necessary for proper cell division, our observations that suspended cells undergo reversible swelling during mitosis suggest that regulation of intracellular water may be a more general component of mitosis than previously appreciated.

  9. Antigen Processing by Autoreactive B Cells Promotes Determinant Spreading

    Institute of Scientific and Technical Information of China (English)

    Yang D.Dai; George Carayanniotis; Eli Sercarz

    2005-01-01

    Acute primary immune responses tend to focus on few immunodominant determinants using a very limited number of T cell clones for expansion, whereas chronic inflammatory responses generally recruit a large number of different T cell clones to attack a broader range of determinants of the invading pathogens or the inflamed tissues.In T cell-mediated organ-specific autoimmune disease, a transition from the acute to the chronic phase contributes to pathogenesis, and the broadening process is called determinant spreading. The cellular components catalyzing the spreading reaction are not identified. It has been suggested that autoreactive B cells may play a central role in diversifying autoreactive T cell responses, possibly through affecting antigen processing and presentation. The clonal identity and diversity of the B cells and antibodies seem critical in regulating T cell activity and subsequent tissue damage or repair. Here, we use two autoimmune animal models, experimental autoimmune thyroiditis (EAT)and type 1 diabetes (T1D), to discuss how autoreactive B cells or antibodies alter the processing and presentation of autoantigens to regulate specific T cell response.

  10. Nanofiber density determines endothelial cell behavior on hydrogel matrix

    Energy Technology Data Exchange (ETDEWEB)

    Berti, Fernanda V., E-mail: fernanda@intelab.ufsc.br [Department of Chemical and Food Engineering, Federal University of Santa Catarina, 88040-900 Florianópolis, SC (Brazil); Rambo, Carlos R. [Department of Electrical Engineering, Federal University of Santa Catarina, 88040-900 Florianópolis, SC (Brazil); Dias, Paulo F. [Department of Cell Biology, Embryology and Genetics, Federal University of Santa Catarina, 88040-900 Florianópolis, SC (Brazil); Porto, Luismar M. [Department of Chemical and Food Engineering, Federal University of Santa Catarina, 88040-900 Florianópolis, SC (Brazil)

    2013-12-01

    When cultured under static conditions, bacterial cellulose pellicles, by the nature of the polymer synthesis that involves molecular oxygen, are characterized by two distinct surface sides. The upper surface is denser in fibers (entangled) than the lower surface that shows greater surface porosity. Human umbilical vein endothelial cells (HUVECs) were used to exploit how the microarchitecture (i.e., surface porosity, fiber network structure, surface topology, and fiber density) of bacterial cellulose pellicle surfaces influence cell–biomaterial interaction and therefore cell behavior. Adhesion, cell ingrowth, proliferation, viability and cell death mechanisms were evaluated on the two pellicle surface sides. Cell behavior, including secondary necrosis, is influenced only by the microarchitecture of the surface, since the biomaterial is extremely pure (constituted of cellulose and water only). Cell–cellulose fiber interaction is the determinant signal in the cell–biomaterial responses, isolated from other frequently present interferences such as protein and other chemical traces usually present in cell culture matrices. Our results suggest that microarchitecture of hydrogel materials might determine the performance of biomedical products, such as bacterial cellulose tissue engineering constructs (BCTECs). - Highlights: • Topography of BC pellicle is relevant to determine endothelial cells' fate. • Cell–biomaterial response is affected by the topography of BC-pellicle surface. • Endothelial cells exhibit different behavior depending on the BC topography. • Apoptosis and necrosis of endothelial cells were affected by the BC topography.

  11. Effect of radiologic contrast media on cell volume regulation in rabbit proximal renal tubules.

    Science.gov (United States)

    Galtung, H K; Løken, M; Sakariassen, K S

    2001-05-01

    Most radiographic contrast media are hyperosmotic and able to shrink cells with which they are in contact. The authors studied cell volume control in rabbit proximal renal tubules after incubation with three contrast media: iohexol, ioxaglate, and iodixanol. Proximal renal tubules were isolated from rabbit kidneys. The tubules were exposed to Ringer solutions containing 5% vol/vol iohexol (final osmolality, 330 mOsm), ioxaglate (323 mOsm), iodixanol (305 mOsm), or mannitol (control solutions with identical osmolalities), and tubule volumes were monitored. After 2 hours of incubation, the tubules were stimulated with a hyposmotic Ringer solution (165 mOsm). Three groups of 10 experiments were performed. All solutions induced cell shrinkage (8.3%+/-3.8 [standard error] to 15.4%+/-0.5), which was completely or partly reversible in most experiments (volume increase, 44.8%+/-14.7 to 149.9%+/-107.3) but not those with iohexol and iodixanol. With exposure to the hyposmotic solution, the cells swelled by 11.0%+/-1.8 to 39.7%+/-4.8. In general, the tubules that had been exposed to the most hyperosmotic solution swelled the most. Those exposed to contrast media showed less swelling than the mannitol-exposed controls. In all control experiments, the cells exhibited a gradual shrinkage (43.6%+/-28.5 to 87.0%+/-13). This regulatory response was partly inhibited in tubules exposed to iohexol (39.9%+/-15.8 shrinkage) or iodixanol (8.9%+/-15.8) and completely inhibited in those exposed to ioxaglate. Iohexol and ioxaglate exposure also led to a decrease in water permeability. Exposure to hyperosmotic contrast medium tends to induce prolonged cell shrinkage, decrease the water permeability of the cellular plasma membranes, and compromise the ability to regulate cellular volume. These changes seem to reflect both the hyperosmolality of the solutions and their inherent chemical properties.

  12. Heteromeric Slick/Slack K+ channels show graded sensitivity to cell volume changes

    DEFF Research Database (Denmark)

    Tejada, Maria A; Hashem, Nadia; Callø, Kirstine

    2017-01-01

    Slick and Slack high-conductance K+ channels are found in the CNS, kidneys, pancreas, among other organs, where they play an important role in cell excitability as well as in ion transport processes. They are both activated by Na+ and Cl- but show a differential regulation by cell volume changes...... to osmotic challenges. In order to provide with the adequate water permeability to the cell membrane of Xenopus laevis oocytes, mRNA of aquaporin 1 was co-expressed with homomeric or heteromeric Slick and Slack α-subunits. Oocytes were superfused with hypotonic or hypertonic buffers and changes in currents...

  13. Life-threatening megaloblastic pancytopenia with normal mean cell volume: case series

    Science.gov (United States)

    Sekhar, Jeevan; Stabler, Sally P.

    2007-01-01

    The mean red blood cell volume (MCV) is usually increased in severe megaloblastic anemia due to pernicious anemia. However, during one year in a university hospital, three patients with life-threatening pancytopenia and normal MCV were proven to have severe vitamin B12 deficiency. The red blood cell distribution width was markedly increased (three times normal) and led to review of the blood smear and recognition of megaloblastosis as well as prominent red cell fragmentation. These three cases illustrate that vitamin B12 status should be evaluated in cases of pancytopenia, independent of the MCV value. PMID:17967337

  14. Formative cell divisions: principal determinants of plant morphogenesis.

    Science.gov (United States)

    Smolarkiewicz, Michalina; Dhonukshe, Pankaj

    2013-03-01

    Formative cell divisions utilizing precise rotations of cell division planes generate and spatially place asymmetric daughters to produce different cell layers. Therefore, by shaping tissues and organs, formative cell divisions dictate multicellular morphogenesis. In animal formative cell divisions, the orientation of the mitotic spindle and cell division planes relies on intrinsic and extrinsic cortical polarity cues. Plants lack known key players from animals, and cell division planes are determined prior to the mitotic spindle stage. Therefore, it appears that plants have evolved specialized mechanisms to execute formative cell divisions. Despite their profound influence on plant architecture, molecular players and cellular mechanisms regulating formative divisions in plants are not well understood. This is because formative cell divisions in plants have been difficult to track owing to their submerged positions and imprecise timings of occurrence. However, by identifying a spatiotemporally inducible cell division plane switch system applicable for advanced microscopy techniques, recent studies have begun to uncover molecular modules and mechanisms for formative cell divisions. The identified molecular modules comprise developmentally triggered transcriptional cascades feeding onto microtubule regulators that now allow dissection of the hierarchy of the events at better spatiotemporal resolutions. Here, we survey the current advances in understanding of formative cell divisions in plants in the context of embryogenesis, stem cell functionality and post-embryonic organ formation.

  15. Involvement of regulatory volume decrease in the migration of nasopharyngeal carcinoma cells

    Institute of Scientific and Technical Information of China (English)

    Jian Wen MAO; Li Xin CHEN; Li Wei WANG; Tim JACOB; Xue Rong SUN; Hui LI; Lin Yan ZHU; Pan LI; Ping ZHONG; Si Huai NIE

    2005-01-01

    The transwell chamber migration assay and CCD digital camera imaging techniques were used to investigate the relationship between regulatory volume decrease (RVD) and cell migration in nasopharyngeal carcinoma cells (CNE-2Z cells). Both migrated and non-migrated CNE-2Z cells, when swollen by 47% hypotonic solution, exhibited RVD which was inhibited by extracellular application of chloride channel blockers adenosine 5'-triphosphate (ATP), 5-nitro-2-(3-phenylpropylamino) benzoic acid (NPPB) and tamoxifen. However, RVD rate in migrated CNE-2Z cells was bigger than that of non-migrated cells and the sensitivity of migrated cells to NPPB and tamoxifen was higher than that of nonmigrated cells. ATP, NPPB and tamoxifen also inhibited migration of CNE-2Z cells. The inhibition of migration was positively correlated to the blockage of RVD, with a correlation coefficient (r) = 0.99, suggesting a functional relationship between RVD and cell migration. We conclude that RVD is involved in cell migration and RVD may play an important role in migratory process in CNE-2Z cells.

  16. Cell cycle-dependent activity of the volume- and Ca2+-activated anion currents in Ehrlich lettre ascites cells

    DEFF Research Database (Denmark)

    Klausen, Thomas Kjaer; Bergdahl, Andreas; Christophersen, Palle

    2007-01-01

    Recent evidence implicates the volume-regulated anion current (VRAC) and other anion currents in control or modulation of cell cycle progression; however, the precise involvement of anion channels in this process is unclear. Here, Cl- currents in Ehrlich Lettre Ascites (ELA) cells were monitored...... during cell cycle progression, under three conditions: (i) after osmotic swelling (i.e., VRAC), (ii) after an increase in the free intracellular Ca2+ concentration (i.e., the Ca2+-activated Cl- current, CaCC), and (iii) under steady-state isotonic conditions. The maximal swelling-activated VRAC current......+ in the pipette), was unaltered from G0 to G1, but decreased in early S phase. A novel high-affinity anion channel inhibitor, the acidic di-aryl-urea NS3728, which inhibited both VRAC and CaCC, attenuated ELA cell growth, suggesting a possible mechanistic link between cell cycle progression and cell cycle...

  17. Uncertainty reevaluation in determining the volume of a silicon sphere by spherical harmonics in an Avogadro project

    Institute of Scientific and Technical Information of China (English)

    Zhang Ji-Tao; Wu Xue-Jian; Li Yan

    2011-01-01

    To determine the Avogadro constant with a target relative uncertainty of 2×10-8,the uncertainty component of the silicon sphere's volume introduced by the spherical harmonics method,which is usually used in determining the sphere's volume,is reevaluated. By means of representing the shape of the silicon sphere by an ellipsoid with Gaussian white noise in its diameters,the uncertainty of the current mapping methods based on the spherical harmonics theory can be estimated theoretically. It is evidenced that the uncertainty component attributed to the current mapping method is underestimated. To eliminate this effect as much as possible,the number of mapping points should be increased to more than before. Moreover,a new mapping method is proposed to accomplish the equal-area mapping with large number points on the silicon sphere.

  18. Determination of turnover and cushion gas volume of a prospected gas storage reservoir under uncertainty

    Energy Technology Data Exchange (ETDEWEB)

    Gubik, A. [RAG-AG Wien (Austria); Baffoe, J.; Schulze-Riegert, R. [SPT Group GmbH, Hamburg (Germany)

    2013-08-01

    Gas storages define a key contribution for building a reliable gas supply chain from production to consumers. In a competitive gas market with short reaction times to seasonal and other gas injection and extraction requirements, gas storages also receive a strong focus on availability and precise prediction estimates for future operation scenarios. Reservoir management workflows are increasingly built on reservoir simulation support for optimizing production schemes and estimating the impact of subsurface uncertainties on field development scenarios. Simulation models for gas storages are calibrated to geological data and accurate reproduction of historical production data are defined as a prerequisite for reliable production and performance forecasts. The underlying model validation process is called history matching, which potentially generates alternative simulation models due to prevailing geological uncertainties. In the past, a single basecase reference model was used to predict production capacities of a gas storage. The working gas volume was precisely defined over a contracted plateau delivery and the required cushion gas volume maintains the reservoir pressure during the operation. Cushion and working gas Volume are strongly dependent on reservoir parameters. In this work an existing depleted gas reservoir and the operation target as a gas storage is described. Key input data to the reservoir model description and simulation is reviewed including production history and geological uncertainties based on large well spacing, limited core and well data and a limited seismic resolution. Target delivery scenarios of the prospected gas storage are evaluated under uncertainty. As one key objective, optimal working gas and cushion gas volumes are described in a probabilistic context reflecting geological uncertainties. Several work steps are defined and included in an integrated workflow design. Equiprobable geological models are generated and evaluated based on

  19. Precise determination of universal finite volume observables in the Gross-Neveu model

    Energy Technology Data Exchange (ETDEWEB)

    Korzec, T.

    2007-01-26

    The Gross-Neveu model is a quantum field theory in two space time dimensions that shares many features with quantum chromo dynamics. In this thesis the continuum model and its discretized versions are reviewed and a finite volume renormalization scheme is introduced and tested. Calculations in the limit of infinitely many fermion flavors as well as perturbative computations are carried out. In extensive Monte-Carlo simulations of the one flavor and the four flavor lattice models with Wilson fermions a set of universal finite volume observables is calculated to a high precision. In the one flavor model which is equivalent to the massless Thirring model the continuum extrapolated Monte-Carlo results are confronted with an exact solution of the model. (orig.)

  20. Enrichment of diluted cell populations from large sample volumes using 3D carbon-electrode dielectrophoresis.

    Science.gov (United States)

    Islam, Monsur; Natu, Rucha; Larraga-Martinez, Maria Fernanda; Martinez-Duarte, Rodrigo

    2016-05-01

    Here, we report on an enrichment protocol using carbon electrode dielectrophoresis to isolate and purify a targeted cell population from sample volumes up to 4 ml. We aim at trapping, washing, and recovering an enriched cell fraction that will facilitate downstream analysis. We used an increasingly diluted sample of yeast, 10(6)-10(2) cells/ml, to demonstrate the isolation and enrichment of few cells at increasing flow rates. A maximum average enrichment of 154.2 ± 23.7 times was achieved when the sample flow rate was 10 μl/min and yeast cells were suspended in low electrically conductive media that maximizes dielectrophoresis trapping. A COMSOL Multiphysics model allowed for the comparison between experimental and simulation results. Discussion is conducted on the discrepancies between such results and how the model can be further improved.

  1. Streaming Model Based Volume Ray Casting Implementation for Cell Broadband Engine

    Directory of Open Access Journals (Sweden)

    Jusub Kim

    2009-01-01

    Full Text Available Interactive high quality volume rendering is becoming increasingly more important as the amount of more complex volumetric data steadily grows. While a number of volumetric rendering techniques have been widely used, ray casting has been recognized as an effective approach for generating high quality visualization. However, for most users, the use of ray casting has been limited to datasets that are very small because of its high demands on computational power and memory bandwidth. However the recent introduction of the Cell Broadband Engine (Cell B.E. processor, which consists of 9 heterogeneous cores designed to handle extremely demanding computations with large streams of data, provides an opportunity to put the ray casting into practical use. In this paper, we introduce an efficient parallel implementation of volume ray casting on the Cell B.E. The implementation is designed to take full advantage of the computational power and memory bandwidth of the Cell B.E. using an intricate orchestration of the ray casting computation on the available heterogeneous resources. Specifically, we introduce streaming model based schemes and techniques to efficiently implement acceleration techniques for ray casting on Cell B.E. In addition to ensuring effective SIMD utilization, our method provides two key benefits: there is no cost for empty space skipping and there is no memory bottleneck on moving volumetric data for processing. Our experimental results show that we can interactively render practical datasets on a single Cell B.E. processor.

  2. Correlation of live-cell imaging with volume scanning electron microscopy.

    Science.gov (United States)

    Lucas, Miriam S; Günthert, Maja; Bittermann, Anne Greet; de Marco, Alex; Wepf, Roger

    2017-01-01

    Live-cell imaging is one of the most widely applied methods in live science. Here we describe two setups for live-cell imaging, which can easily be combined with volume SEM for correlative studies. The first procedure applies cell culture dishes with a gridded glass support, which can be used for any light microscopy modality. The second approach is a flow-chamber setup based on Ibidi μ-slides. Both live-cell imaging strategies can be followed up with serial blockface- or focused ion beam-scanning electron microscopy. Two types of resin embedding after heavy metal staining and dehydration are presented making best use of the particular advantages of each imaging modality: classical en-bloc embedding and thin-layer plastification. The latter can be used only for focused ion beam-scanning electron microscopy, but is advantageous for studying cell-interactions with specific substrates, or when the substrate cannot be removed. En-bloc embedding has diverse applications and can be applied for both described volume scanning electron microscopy techniques. Finally, strategies for relocating the cell of interest are discussed for both embedding approaches and in respect to the applied light and scanning electron microscopy methods. Copyright © 2017 Elsevier Inc. All rights reserved.

  3. Optical quantification of cellular mass, volume and density of circulating tumor cells identified in an ovarian cancer patient

    Directory of Open Access Journals (Sweden)

    Kevin Gregory Phillips

    2012-07-01

    Full Text Available Clinical studies have demonstrated that circulating tumor cells (CTCs are present in the blood of cancer patients with known metastatic disease across the major types of epithelial malignancies. Recent studies have shown that the concentration of CTCs in the blood is prognostic of overall survival in breast, prostate, colorectal and non-small cell lung cancer. This study characterizes CTCs identified using the high-definition (HD-CTC assay in an ovarian cancer patient with stage IIIC disease. We characterized the physical properties of 31 HD-CTCs and 50 normal leukocytes from a single blood draw taken just prior to the initial debulking surgery. We utilized a non-interferometric quantitative phase microscopy technique using brightfield imagery to measure cellular dry mass. Next we used a quantitative differential interference contrast microscopy technique to measure cellular volume. These techniques were combined to determine cellular dry mass density. We found that HD-CTCs were more massive than leukocytes: 33.6 ± 3.2 pg (HD-CTC compared to 18.7 ± 0.6 pg (leukocytes, p < 0.001; had greater volumes: 518.3 ± 24.5 fL (HD-CTC compared to 230.9 ± 78.5 fL (leukocyte, p<0.001; and possessed a decreased dry mass density with respect to leukocytes: 0.065 ± 0.006 pg/fL (HD-CTC compared to 0.085 ± 0.004 pg/fL (leukocyte, p < 0.006. Quantification of HD-CTC dry mass content and volume provide key insights into the fluid dynamics of cancer, and may provide the rationale for strategies to isolate, monitor or target CTCs based on their physical properties. The parameters reported here can also be incorporated into blood cell flow models to better understand metastasis.

  4. Current Injection Provokes Rapid Expansion of the Guard Cell Cytosolic Volume and Triggers Ca(2+) Signals.

    Science.gov (United States)

    Voss, Lena J; Hedrich, Rainer; Roelfsema, M Rob G

    2016-03-07

    High-resolution microscopy opens the door for detailed single-cell studies with fluorescent reporter dyes and proteins. We used a confocal spinning disc microscope to monitor fluorescent dyes and the fluorescent protein Venus in tobacco and Arabidopsis guard cells. Multi-barreled microelectrodes were used to inject dyes and apply voltage pulses, which provoke transient rises in the cytosolic Ca(2+) level. Voltage pulses also caused changes in the distribution of Lucifer Yellow and Venus, which pointed to a reversible increase of guard cell cytosolic volume. The dynamic cytosolic volume changes turned out to be provoked by current injection of ions. A reduction of the clamp current, by blocking K(+) uptake channels with Cs(+), strongly suppressed the cytosolic volume changes. Cs(+) not only inhibited the expansion of the cytosol, but also inhibited hyperpolarization-induced elevations of the cytosolic Ca(2+) concentration. A complete loss of voltage-induced Ca(2+) signals occurred when Ca(2+)-permeable plasma membrane channels were simultaneously blocked with La(3+). This shows that two mechanisms cause hyperpolarization-induced elevation of the cytosolic Ca(2+)-concentration: (i) activation of voltage-dependent Ca(2+)-permeable channels, (ii) osmotically induced expansion of the cytosol, which leads to a release of Ca(2+) from intracellular stores.

  5. Synapse loss from chronically elevated glucocorticoids: relationship to neuropil volume and cell number in hippocampal area CA3.

    Science.gov (United States)

    Tata, Despina A; Marciano, Veronica A; Anderson, Brenda J

    2006-09-20

    Individuals with clinical disorders associated with elevated plasma glucocorticoids, such as major depressive disorder and Cushing's syndrome, are reported to have smaller hippocampal volume. To understand how the hippocampus responds at the cellular and subcellular levels to glucocorticoids and how such changes are related to volume measures, we have undertaken a comprehensive study of glucocorticoid effects on hippocampal CA3 volume and identified elements in the neuropil including astrocytic volume and cell and synapse number and size. Male Sprague-Dawley rats were injected with corticosterone (40 mg/kg), the primary glucocorticoid in rodents, or vehicle for 60 days. The CA3 was further subdivided so that the two-thirds of CA3 (nearest the dentate gyrus) previously shown to be vulnerable to corticosterone could be analyzed as two separate subfields. Corticosterone had no effect on neuropil volume or glial volume in the proximal subfield but caused a strong tendency for astrocytic processes to make up a larger proportion of the tissue and for volume of tissue made of constituents other than glial cells (primarily neuronal processes) to be smaller in the middle subfield. Within the neuropil, there were no cellular or subcellular profiles that indicated degeneration, suggesting that corticosterone does not cause prolonged damage. Corticosterone did not reduce cell number or cell or nonperforated synapse size but did cause a pronounced loss of synapses. This loss occurred in both subfields and, therefore, was independent of volume loss. Together, the findings suggest that volume measures can underestimate corticosterone effects on neural structure.

  6. Cell fate determination by ubiquitin-dependent regulation of translation

    Science.gov (United States)

    Werner, Achim; Iwasaki, Shintaro; McGourty, Colleen; Medina-Ruiz, Sofia; Teerikorpi, Nia; Fedrigo, Indro; Ingolia, Nicholas T.; Rape, Michael

    2015-01-01

    Metazoan development depends on accurate execution of differentiation programs that allow pluripotent stem cells to adopt specific fates 1. Differentiation requires changes to chromatin architecture and transcriptional networks, yet whether other regulatory events support cell fate determination is less well understood. Here, we have identified the vertebrate-specific ubiquitin ligase CUL3KBTBD8 as an essential regulator of neural crest specification. CUL3KBTBD8 monoubiquitylates NOLC1 and its paralog TCOF1, whose mutation underlies the neurocristopathy Treacher Collins Syndrome 2,3. Ubiquitylation drives formation of a TCOF1-NOLC1 platform that connects RNA polymerase I with ribosome modification enzymes and remodels the translational program of differentiating cells in favor of neural crest specification. We conclude that ubiquitin-dependent regulation of translation is an important feature of cell fate determination. PMID:26399832

  7. Convergence of Cell Based Finite Volume Discretizations for Problems of Control in the Conduction Coefficients

    DEFF Research Database (Denmark)

    Evgrafov, Anton; Gregersen, Misha Marie; Sørensen, Mads Peter

    2011-01-01

    design, in particular shape and topology optimization, and are most often solved numerically utilizing a finite element approach. Within the FV framework for control in the coefficients problems the main difficulty we face is the need to analyze the convergence of fluxes defined on the faces of cells......We present a convergence analysis of a cell-based finite volume (FV) discretization scheme applied to a problem of control in the coefficients of a generalized Laplace equation modelling, for example, a steady state heat conduction. Such problems arise in applications dealing with geometric optimal...

  8. Determination of Internal Target Volume for Radiation Treatment Planning of Esophageal Cancer by Using 4-Dimensional Computed Tomography (4DCT)

    Energy Technology Data Exchange (ETDEWEB)

    Chen, Xiaojian [Department of Radiation Oncology, Medical College of Wisconsin, Milwaukee, Wisconsin (United States); Lu, Haijun [Department of Radiation Oncology, Medical College of Wisconsin, Milwaukee, Wisconsin (United States); Radiation Oncology Center, Affiliated Hospital of Medical College, Qingdao University, Qingdao (China); Tai, An; Johnstone, Candice; Gore, Elizabeth [Department of Radiation Oncology, Medical College of Wisconsin, Milwaukee, Wisconsin (United States); Li, X. Allen, E-mail: ali@mcw.edu [Department of Radiation Oncology, Medical College of Wisconsin, Milwaukee, Wisconsin (United States)

    2014-09-01

    Purpose: To determine an efficient strategy for the generation of the internal target volume (ITV) for radiation treatment planning for esophageal cancer using 4-dimensional computed tomography (4DCT). Methods and Materials: 4DCT sets acquired for 20 patients with esophageal carcinoma were analyzed. Each of the 4DCT sets was binned into 10 respiratory phases. For each patient, the gross tumor volume (GTV) was delineated on the 4DCT set at each phase. Various strategies to derive ITV were explored, including the volume from the maximum intensity projection (MIP; ITV{sub M}IP), unions of the GTVs from selected multiple phases ITV2 (0% and 50% phases), ITV3 (ITV2 plus 80%), and ITV4 (ITV3 plus 60%), as well as the volumes expanded from ITV2 and ITV3 with a uniform margin. These ITVs were compared to ITV10 (the union of the GTVs for all 10 phases) and the differences were measured with the overlap ratio (OR) and relative volume ratio (RVR) relative to ITV10 (ITVx/ITV10). Results: For all patients studied, the average GTV from a single phase was 84.9% of ITV10. The average ORs were 91.2%, 91.3%, 94.5%, and 96.4% for ITV{sub M}IP, ITV2, ITV3, and ITV4, respectively. Low ORs were associated with irregular breathing patterns. ITV3s plus 1 mm uniform margins (ITV3+1) led to an average OR of 98.1% and an average RVR of 106.4%. Conclusions: The ITV generated directly from MIP underestimates the range of the respiration motion for esophageal cancer. The ITV generated from 3 phases (ITV3) may be used for regular breathers, whereas the ITV generated from 4 phases (ITV4) or ITV3 plus a 1-mm uniform margin may be applied for irregular breathers.

  9. Biomaterial surface proteomic signature determines interaction with epithelial cells.

    Science.gov (United States)

    Abdallah, Mohamed-Nur; Tran, Simon D; Abughanam, Ghada; Laurenti, Marco; Zuanazzi, David; Mezour, Mohamed A; Xiao, Yizhi; Cerruti, Marta; Siqueira, Walter L; Tamimi, Faleh

    2017-03-01

    Cells interact with biomaterials indirectly through extracellular matrix (ECM) proteins adsorbed onto their surface. Accordingly, it could be hypothesized that the surface proteomic signature of a biomaterial might determine its interaction with cells. Here, we present a surface proteomic approach to test this hypothesis in the specific case of biomaterial-epithelial cell interactions. In particular, we determined the surface proteomic signature of different biomaterials exposed to the ECM of epithelial cells (basal lamina). We revealed that the biomaterial surface chemistry determines the surface proteomic profile, and subsequently the interaction with epithelial cells. In addition, we found that biomaterials with surface chemistries closer to that of percutaneous tissues, such as aminated PMMA and aminated PDLLA, promoted higher selective adsorption of key basal lamina proteins (laminins, nidogen-1) and subsequently improved their interactions with epithelial cells. These findings suggest that mimicking the surface chemistry of natural percutaneous tissues can improve biomaterial-epithelial integration, and thus provide a rationale for the design of improved biomaterial surfaces for skin regeneration and percutaneous medical devices.

  10. Volume regulation of intestinal cells of echinoderms: Putative role of ion transporters (Na(+)/K(+)-ATPase and NKCC).

    Science.gov (United States)

    Castellano, Giovanna C; Souza, Marta M; Freire, Carolina A

    2016-11-01

    Echinoderms are exclusively marine osmoconformer invertebrates. Some species occupy the challenging intertidal region. Upon salinity changes, the extracellular osmotic concentration of these animals also varies, exposing tissues and cells to osmotic challenges. Cells and tissues may then respond with volume regulation mechanisms, which involve transport of ions and water into and/or out of the cells, through ion transporters, such as the Na(+)/K(+)-ATPase and NKCC. The goal of this study was to relate the cell volume regulation capacity of echinoderm intestinal cells Na(+)/K(+)-ATPase and NKCC activities, in three echinoderm species: Holothuria grisea, Arbacia lixula, and Echinometra lucunter. Isolated cells of these species displayed some control of their cell volume upon exposure to anisosmotic media (isolated intestinal cells, calcein fluorescence as indicator of volume change), with a distinct higher capacity shown by H. grisea, which did not swell even upon 50% hyposmotic shock. The holothuroid cells showed indirect evidence (effect of furosemide) of the participation of NKCC in this process, with a secretory function, and of a secondary role by the NKA (effect of ouabain). Other mechanisms are probably responsible for this function in the urchins. Variable expression of these transporters, and others not examined here, may to some extent account for the variability in cell volume regulation capacity in echinoderm cells.

  11. Determination of thermal properties of commercial Ni-MH cells

    Science.gov (United States)

    Darcy, Eric C.

    1994-01-01

    The test objectives were to evaluate the electrical and thermal performance of commercial Ni-MH cells, evaluate the effectiveness of commercial charge control circuits, assess the abuse tolerance of these cells, and correlate performance and abuse tolerances to cell design via disassembly. Design objectives were to determine which cell designs are most suitable for scale-up and to guide the design of future shuttle and space station based battery chargers. Results, displayed in viewgraph format, include: reflex charging with ICS circuit resulted in premature charge termination; Ni-MH cells appear very tolerant to overcharge at low rates; Enstore's charger is more electrically and thermally efficient at high rates; and Ni-MH cycles much more efficiently than Ni-Cd with the delta-V/delta-t termination.

  12. Determinism and probability in the development of the cell theory.

    Science.gov (United States)

    Duchesneau, François

    2012-09-01

    A return to Claude Bernard's original use of the concept of 'determinism' displays the fact that natural laws were presumed to rule over all natural processes. In a more restricted sense, the term boiled down to a mere presupposition of constant determinant causes for those processes, leaving aside any particular ontological principle, even stochastic. The history of the cell theory until around 1900 was dominated by a twofold conception of determinant causes. Along a reductionist trend, cells' structures and processes were supposed to be accounted for through their analysis into detailed partial mechanisms. But a more holistic approach tended to subsume those analytic means and the mechanism involved under a program of global functional determinations. When mitotic and meiotic sequences in nuclear replication were being unveiled and that neo-Mendelian genetics was being grafted onto cytology and embryology, a conception of strict determinism at the nuclear level, principally represented by Wilhelm Roux and August Weismann, would seem to rule unilaterally over the mosaic interpretation of the cleavage of blastomeres. But, as shown by E.B. Wilson, in developmental processes there occur contingent outcomes of cell division which observations and experiments reveal. This induces the need to admit 'epigenetic' determinants and relativize the presumed 'preformation' of thedevelopmental phases by making room for an emergent order which the accidental circumstances of gene replication would trigger on.

  13. Small Cell Lung Cancer: The Influence of Dose and Treatment Volume on Outcome.

    Science.gov (United States)

    Lichter; Turrisi

    1995-01-01

    The treatment of small cell lung cancer is clearly enhanced by the addition of radiation therapy. Survival increases modestly while local thoracic failure as first site of progression is reduced from approximately 60% when chemotherapy alone is used to 30% after combined modality therapy. The variables of radiation dose and treatment volume seem to be important in the successful management of this disease. Local chest control appears to increase as doses are escalated from low levels (25 Gy) to moderate levels (45 to 50 Gy(. With about one third of patients experiencing local chest progression, one can speculate that higher radiation doses might be of value. However, at this time there is no proof that increased dose or dose intensity bears out this promise. Indeed, increasing dose intensity of radiotherapy, eg, twice-daily treatment, increases esophagitis, perhaps reduces local failure, but has not improved overall survival. Using larger total doses or altered fraction schemes must still be considered to be under investigation. To increase dose in a safe manner, reduction in the volume covered by radiation portals will likely need to take place. Modern trials suggest that prophylactic treatment of the radiographically or clinically negative contralateral hilum and/or supraclavicular nodal regions may not be necessary for survival or local control. Importantly, reducing treatment volumes may permit increasing doses without exceding normal tissue tolerance. Also, reduced volumes pave the way for further clinical trials that improve radiation dose delivery by better target definition and more conformal therapy.

  14. Determination of Energy Characteristic and Microporous Volume by Immersion Calorimetry in Carbon Monoliths

    Directory of Open Access Journals (Sweden)

    Juan Carlos Moreno-Piraján

    2012-01-01

    Full Text Available Activated carbon monoliths disc and honeycomb type were prepared by chemical activation of coconut shell with zinc chloride at different concentrations, without using a binder. The structures were characterized by N2 adsorption at 77 K and immersion calorimetry into benzene. The experimental results showed that the activation with zinc chloride produces a wide microporous development, with micropore volume between 0,38 and 0,79 cm3g-1, apparent BET surface area between 725 and 1523 m2g-1 and immersion enthalpy between 73,5 and 164,2 Jg-1. We compared the experimental enthalpy with calculated enthalpy by equation Stoeckli-Kraehenbuehl finding a data dispersion from which can infer that the structures are not purely microporous; this fact is ratified with similar behavior that the evidence t the product EoWo.

  15. Galvanic Cells and the Determination of Equilibrium Constants

    Science.gov (United States)

    Brosmer, Jonathan L.; Peters, Dennis G.

    2012-01-01

    Readily assembled mini-galvanic cells can be employed to compare their observed voltages with those predicted from the Nernst equation and to determine solubility products for silver halides and overall formation constants for metal-ammonia complexes. Results obtained by students in both an honors-level first-year course in general chemistry and…

  16. Galvanic Cells and the Determination of Equilibrium Constants

    Science.gov (United States)

    Brosmer, Jonathan L.; Peters, Dennis G.

    2012-01-01

    Readily assembled mini-galvanic cells can be employed to compare their observed voltages with those predicted from the Nernst equation and to determine solubility products for silver halides and overall formation constants for metal-ammonia complexes. Results obtained by students in both an honors-level first-year course in general chemistry and…

  17. Impaired Cell Volume Regulation in Intestinal Crypt Epithelia of Cystic Fibrosis Mice

    Science.gov (United States)

    Valverde, M. A.; O'Brien, J. A.; Sepulveda, F. V.; Ratcliff, R. A.; Evans, M. J.; Colledge, W. H.

    1995-09-01

    Cystic fibrosis is a disease characterized by abnormalities in the epithelia of the lungs, intestine, salivary and sweat glands, liver, and reproductive systems, often as a result of inadequate hydration of their secretions. The primary defect in cystic fibrosis is the altered activity of a cAMP-activated Cl^- channel, the cystic fibrosis transmembrane conductance regulator (CFTR) channel. However, it is not clear how a defect in the CFTR Cl^- channel function leads to the observed pathological changes. Although much is known about the structural properties and regulation of the CFTR, little is known of its relationship to cellular functions other than the cAMP-dependent Cl^- secretion. Here we report that cell volume regulation after hypotonic challenge is also defective in intestinal crypt epithelial cells isolated from CFTR -/- mutant mice. Moreover, the impairment of the regulatory volume decrease in CFTR -/- crypts appears to be related to the inability of a K^+ conductance to provide a pathway for the exit of this cation during the volume adjustments. This provides evidence that the lack of CFTR protein may have additional consequences for the cellular function other than the abnormal cAMP-mediated Cl^- secretion.

  18. A large volume cell for in situ neutron diffraction studies of hydrothermal crystallizations

    Science.gov (United States)

    Xia, Fang; Qian, Gujie; Brugger, Joël; Studer, Andrew; Olsen, Scott; Pring, Allan

    2010-10-01

    A hydrothermal cell with 320 ml internal volume has been designed and constructed for in situ neutron diffraction studies of hydrothermal crystallizations. The cell design adopts a dumbbell configuration assembled with standard commercial stainless steel components and a zero-scattering Ti-Zr alloy sample compartment. The fluid movement and heat transfer are simply driven by natural convection due to the natural temperature gradient along the fluid path, so that the temperature at the sample compartment can be stably sustained by heating the fluid in the bottom fluid reservoir. The cell can operate at temperatures up to 300 °C and pressures up to 90 bars and is suitable for studying reactions requiring a large volume of hydrothermal fluid to damp out the negative effect from the change of fluid composition during the course of the reactions. The capability of the cell was demonstrated by a hydrothermal phase transformation investigation from leucite (KAlSi2O6) to analcime (NaAlSi2O6ṡH2O) at 210 °C on the high intensity powder diffractometer Wombat in ANSTO. The kinetics of the transformation has been resolved by collecting diffraction patterns every 10 min followed by Rietveld quantitative phase analysis. The classical Avrami/Arrhenius analysis gives an activation energy of 82.3±1.1 kJ mol-1. Estimations of the reaction rate under natural environments by extrapolations agree well with petrological observations.

  19. Sendai virus utilizes specific sialyloligosaccharides as host cell receptor determinants.

    Science.gov (United States)

    Markwell, M A; Paulson, J C

    1980-10-01

    Purified sialyltransferases (CMP-N-acetyl-neuraminate:D-galactosyl-glycoprotein N-acetylneuraminyl-transferase, EC 2.4.99.1) in conjunction with neuraminidase (acylneuraminyl hydrolase, EC 3.2.1.18) were used to produce cell surface sialyloligosaccharides of defined sequence to investigate their role in paramyxovirus infection of host cells. Infection of Madin-Darby bovine kidney cells by Sendai virus was monitored by hemagglutination titer of the virus produced and by changes in morphological characteristics. By either criterion, treatment of the cells with Vibrio cholerae neuraminidase to remove cell surface sialic acids rendered them resistant to infection by Sendai virus. Endogenous replacement of receptors by the cell occurred slowly but supported maximal levels of infection within 6 hr. In contrast, sialylation during a 20-min incubation with CMP-sialic acid and beta-galactoside alpha 2,3-sialytransferase restored full susceptibility to infection. This enzyme elaborates the NeuAc alpha 2,3Gal beta 1,3GalNAc (NeuAc, N-acetylneuraminic acid) sequence on glycoproteins and glycolipids. No restoration of infectivity was observed when neuraminidase-treated cells were sialylated by using beta-galactoside alpha 2,6-sialytransferase, which elaborates the NeuAc-alpha 2,6Gal beta 1,4GlcNAc sequence. These results suggest that sialyloligosaccharide receptor determinants of defined sequence are required for Sendai virus infection of host cells.

  20. Effect of radiologic contrast material on cell volume regulation in proximal renal tubules from trout (Salmo trutta).

    Science.gov (United States)

    Galtung, H K; Løken, M; Sakariassen, K S

    2000-11-01

    Most radiographic contrast media (CM) are hyperosmotic and pose an osmotic threat to cells they are in contact with. To study these effects at the cellular level, cell volume regulatory mechanisms were observed in proximal renal tubules following exposure to the CM iohexol, ioxaglate, and iodixanol. Isolated renal tubules from trout (Salmo trutta) were exposed to 5% vol/vol iohexol (326 mOsm), ioxaglate (314 mOsm), or iodixanol (300 mOsm) or mannitol (to achieve the same osmolalities), and cell volume changes were observed videometrically. Iohexol and ioxaglate solutions induced a rapid shrinkage (12%-13%) not followed by cell volume regulation. Without CM (same osmolality), the cells shrank 11% but then showed a 77%-88% volume recovery. This reswelling was inhibited by 55% with the Na+, K+, Cl- symporter inhibitor bumetanide (50 micromol/L). Iodixanol did not significantly affect cell volume. Tubules preincubated with CM or mannitol were then stimulated with a hypoosmotic Ringer solution (160 mOsm) resulting in a 26%-36% cellular volume increase. Compared with results of experiments without mannitol and CM, preexposure to iohexol or ioxaglate almost completely inhibited the expected regulatory shrinkage phase, while previous exposure to hyperosmotic solutions with mannitol reduced the shrinkage response by 40%-53%. In this system, the hyperosmotic iohexol and ioxaglate cause cell shrinkage followed by an impaired cell volume regulatory response. Exposure to these two CM also inhibits cell volume regulation on hypoosmotic stimulation. The isosmotic iodixanol has no such effects. These changes appear to some extent to be a result of the CM's degree of hyperosmolality, but this property alone does not explain these findings.

  1. Direct determination of phosphatase activity from physiological substrates in cells.

    Directory of Open Access Journals (Sweden)

    Zhongyuan Ren

    Full Text Available A direct and continuous approach to determine simultaneously protein and phosphate concentrations in cells and kinetics of phosphate release from physiological substrates by cells without any labeling has been developed. Among the enzymes having a phosphatase activity, tissue non-specific alkaline phosphatase (TNAP performs indispensable, multiple functions in humans. It is expressed in numerous tissues with high levels detected in bones, liver and neurons. It is absolutely required for bone mineralization and also necessary for neurotransmitter synthesis. We provided the proof of concept that infrared spectroscopy is a reliable assay to determine a phosphatase activity in the osteoblasts. For the first time, an overall specific phosphatase activity in cells was determined in a single step by measuring simultaneously protein and substrate concentrations. We found specific activities in osteoblast like cells amounting to 116 ± 13 nmol min(-1 mg(-1 for PPi, to 56 ± 11 nmol min(-1 mg(-1 for AMP, to 79 ± 23 nmol min(-1 mg(-1 for beta-glycerophosphate and to 73 ± 15 nmol min(-1 mg(-1 for 1-alpha-D glucose phosphate. The assay was also effective to monitor phosphatase activity in primary osteoblasts and in matrix vesicles. The use of levamisole--a TNAP inhibitor--served to demonstrate that a part of the phosphatase activity originated from this enzyme. An IC50 value of 1.16 ± 0.03 mM was obtained for the inhibition of phosphatase activity of levamisole in osteoblast like cells. The infrared assay could be extended to determine any type of phosphatase activity in other cells. It may serve as a metabolomic tool to monitor an overall phosphatase activity including acid phosphatases or other related enzymes.

  2. Determination of the steam volume fraction in the event of loss of cooling of the spent fuel storage pool

    Science.gov (United States)

    Sledkov, R. M.; Galkin, I. Yu.; Stepanov, O. E.; Strebnev, N. A.

    2017-01-01

    When one solves engineering problems related to the cooling of fuel assemblies (FAs) in a spent fuel storage pool (SFSP) and the assessment of nuclear safety of FA storage in an SFSP in the initial event of loss of SFSP cooling, it is essential to determine the coolant density and, consequently, steam volume fractions φ in bundles of fuel elements at a pressure of 0.1-0.5 MPa. Such formulas for calculating φ that remain valid in a wide range of operating parameters and geometric shapes of channels and take the conditions of loss of SFSP cooling into account are currently almost lacking. The results of systematization and analysis of the available formulas for φ are reported in the present study. The calculated values were compared with the experimental data obtained in the process of simulating the conditions of FA cooling in an SFSP in the event of loss of its cooling. Six formulas for calculating the steam volume fraction, which were used in this comparison, were chosen from a total of 11 considered relations. As a result, the formulas producing the most accurate values of φ in the conditions of loss of SFSP cooling were selected. In addition, a relation that allows one to perform more accurate calculations of steam volume fractions in the conditions of loss of SFSP cooling was derived based on the Fedorov formula in the two-group approximation.

  3. Evaluation of a Novel Approach for Automatic Volume Determination of Glioblastomas Based on Several Manual Expert Segmentations

    CERN Document Server

    Egger, Jan; Kuhnt, Daniela; Carl, Barbara; Kappus, Christoph; Freisleben, Bernd; Nimsky, Christopher

    2011-01-01

    The glioblastoma multiforme is the most common malignant primary brain tumor and is one of the highest malignant human neoplasms. During the course of disease, the evaluation of tumor volume is an essential part of the clinical follow-up. However, manual segmentation for acquisition of tumor volume is a time-consuming process. In this paper, a new approach for the automatic segmentation and volume determination of glioblastomas (glioblastoma multiforme) is presented and evaluated. The approach uses a user-defined seed point inside the glioma to set up a directed 3D graph. The nodes of the graph are obtained by sampling along rays that are sent through the surface points of a polyhedron. After the graph has been constructed, the minimal s-t cut is calculated to separate the glioblastoma from the background. For evaluation, 12 Magnetic Resonance Imaging (MRI) data sets were manually segmented slice by slice, by neurosurgeons with several years of experience in the resection of gliomas. Afterwards, the manual se...

  4. Determination of respirable mass concentration using a high volume air sampler and a sedimentation method for fractionation

    Energy Technology Data Exchange (ETDEWEB)

    Johnson, J.

    1995-12-31

    A preliminary study of a new method for determining respirable mass concentration is described. This method uses a high volume air sampler and subsequent fractionation of the collected mass using a particle sedimentation technique. Side-by-side comparisons of this method with cyclones were made in the field and in the laboratory. There was good agreement among the samplers in the laboratory, but poor agreement in the field. The effect of wind on the samplers` capture efficiencies is the primary hypothesized source of error among the field results. The field test took place at the construction site of a hazardous waste landfill located on the Hanford Reservation.

  5. Analytical challenges of determining composition and structure in small volumes with applications to semiconductor technology, nanostructures and solid state science

    Science.gov (United States)

    Ma, Zhiyong; Kuhn, Markus; Johnson, David C.

    2017-03-01

    Determining the structure and composition of small volumes is vital to the ability to understand and control nanoscale properties and critical for advancing both fundamental science and applications, such as semiconductor device manufacturing. While metrology of nanoscale materials (nanoparticles, nanocomposites) and nanoscale semiconductor structures is challenging, both basic research and cutting edge technology benefit from new and enhanced analytical techniques. This focus issue contains articles describing approaches to overcome the challenges in obtaining statistically significant atomic-scale quantification of structure and composition in a variety of materials and devices using electron microscopy and atom probe tomography.

  6. Condition-dependent cell volume and concentration of Escherichia coli to facilitate data conversion for systems biology modeling

    NARCIS (Netherlands)

    Volkmer, Benjamin; Heinemann, Matthias

    2011-01-01

    Systems biology modeling typically requires quantitative experimental data such as intracellular concentrations or copy numbers per cell. In order to convert population-averaging omics measurement data to intracellular concentrations or cellular copy numbers, the total cell volume and number of cell

  7. Robust high-performance nanoliter-volume single-cell multiple displacement amplification on planar substrates

    Science.gov (United States)

    Leung, Kaston; Klaus, Anders; Lin, Bill K.; Laks, Emma; Biele, Justina; Lai, Daniel; Bashashati, Ali; Huang, Yi-Fei; Aniba, Radhouane; Moksa, Michelle; Steif, Adi; Mes-Masson, Anne-Marie; Hirst, Martin; Shah, Sohrab P.; Aparicio, Samuel; Hansen, Carl L.

    2016-01-01

    The genomes of large numbers of single cells must be sequenced to further understanding of the biological significance of genomic heterogeneity in complex systems. Whole genome amplification (WGA) of single cells is generally the first step in such studies, but is prone to nonuniformity that can compromise genomic measurement accuracy. Despite recent advances, robust performance in high-throughput single-cell WGA remains elusive. Here, we introduce droplet multiple displacement amplification (MDA), a method that uses commercially available liquid dispensing to perform high-throughput single-cell MDA in nanoliter volumes. The performance of droplet MDA is characterized using a large dataset of 129 normal diploid cells, and is shown to exceed previously reported single-cell WGA methods in amplification uniformity, genome coverage, and/or robustness. We achieve up to 80% coverage of a single-cell genome at 5× sequencing depth, and demonstrate excellent single-nucleotide variant (SNV) detection using targeted sequencing of droplet MDA product to achieve a median allelic dropout of 15%, and using whole genome sequencing to achieve false and true positive rates of 9.66 × 10−6 and 68.8%, respectively, in a G1-phase cell. We further show that droplet MDA allows for the detection of copy number variants (CNVs) as small as 30 kb in single cells of an ovarian cancer cell line and as small as 9 Mb in two high-grade serous ovarian cancer samples using only 0.02× depth. Droplet MDA provides an accessible and scalable method for performing robust and accurate CNV and SNV measurements on large numbers of single cells. PMID:27412862

  8. A Volume-Weighting Cloud-in-Cell Model for Particle Simulation of Axially Symmetric Plasmas

    Institute of Scientific and Technical Information of China (English)

    李永东; 何锋; 刘纯亮

    2005-01-01

    A volume-weighting cloud-in-cell (VW-CIC) model is developed to implement the particle-in-cell (PIC) simulation in axially symmetric systems. This model gives a first-order accuracy in the cylindrical system, and it is incorporated into a PIC code. A planar diode with a finite-radius circular emitter is simulated with the code. The simulation results show that the VW-CIC model has a better accuracy and a lower noise than the conventional area-weighting cloud-in-cell (AW-CIC) model, especially on those points near the axis. The two-dimensional (2-D) space-charge-limited current density obtained from VW-CIC model is in better agreement with Lau's analytical result. This model is more suitable for 2.5-D PIC simulation of axially symmetric plasmas.

  9. Mimetic Theory for Cell-Centered Lagrangian Finite Volume Formulation on General Unstructured Grids

    Energy Technology Data Exchange (ETDEWEB)

    Sambasivan, Shiv Kumar [Los Alamos National Laboratory; Shashkov, Mikhail J. [Los Alamos National Laboratory; Burton, Donald E. [Los Alamos National Laboratory; Christon, Mark A. [Los Alamos National Laboratory

    2012-07-19

    A finite volume cell-centered Lagrangian scheme for solving large deformation problems is constructed based on the hypo-elastic model and using the mimetic theory. Rigorous analysis in the context of gas and solid dynamics, and arbitrary polygonal meshes, is presented to demonstrate the ability of cell-centered schemes in mimicking the continuum properties and principles at the discrete level. A new mimetic formulation based gradient evaluation technique and physics-based, frame independent and symmetry preserving slope limiters are proposed. Furthermore, a physically consistent dissipation model is employed which is both robust and inexpensive to implement. The cell-centered scheme along with these additional new features are applied to solve solids undergoing elasto-plastic deformation.

  10. Inhibition of volume-activated chloride currents in endothelial cells by chromones.

    Science.gov (United States)

    Heinke, S.; Szücs, G.; Norris, A.; Droogmans, G.; Nilius, B.

    1995-01-01

    1. We have studied the effects of the reported chloride channel blocker, sodium cromoglycate, on volume-activated Cl- currents in endothelial cells from bovine pulmonary artery by means of the whole-cell patch clamp technique. Cl- currents were activated by challenging the cells with a hypotonic extracellular solution of 60% of the normal osmolarity. 2. Half maximal activation of the current at +95 mV occurred after exposure of the cells for 148 +/- 10 s (n = 6) to hypotonic solution (HTS). At the same membrane potential but in the presence of 100 microM sodium cromoglycate (disodium-1,3-bis (2'-carboxylate-chromone-5'-yloxy)-2-hydroxy-propane) activation was delayed (253 +/- 25 s, n = 6) and the maximal current amplitude was reduced to 63 +/- 7% of the control (n = 13). 3. In comparison, an equimolar concentration of NPPB (5-nitro-2(3-phenyl) propylamino-benzoic acid), another Cl- channel blocker, completely blocked the volume-activated current in less than 20 s. 4. Sodium cromoglycate, applied at the time when the HTS-induced current was completely activated, dose-dependently inhibited this current with a concentration for half maximal inhibition of 310 +/- 70 microM. Data for nedocromil sodium were not significantly different from those for sodium cromoglycate. 5. Sodium cromoglycate, loaded into the endothelial cells via the patch pipette in ruptured patches, resulted in a decline of the HTS activated current with a time course that was compatible with diffusion of the compound from the pipette into the cell.(ABSTRACT TRUNCATED AT 250 WORDS) PMID:8564197

  11. Reliability and validity of non-invasive determined haemoglobin mass and blood volumes

    DEFF Research Database (Denmark)

    Fagoni, Nazzareno; Andersen, Andreas Breenfeldt; Oberholzer, Laura

    2017-01-01

    INTRODUCTION: The carbon monoxide (CO) rebreathing method used for the determination of haemoglobin mass (Hbmass ) is associated with blood sample analysis (in this study: Radiometer ABL800). As an alternative hereto the aim of the present study was to evaluate the use of a portable and non-invas...

  12. Cell volume control in phospholemman (PLM) knockout mice: do cardiac myocytes demonstrate a regulatory volume decrease and is this influenced by deletion of PLM?

    Science.gov (United States)

    Bell, James R; Lloyd, David; Curl, Claire L; Delbridge, Lea M D; Shattock, Michael J

    2009-03-01

    In addition to modulatory actions on Na+-K+-ATPase, phospholemman (PLM) has been proposed to play a role in cell volume regulation. Overexpression of PLM induces ionic conductances, with 'PLM channels' exhibiting selectivity for taurine. Osmotic challenge of host cells overexpressing PLM increases taurine efflux and augments the cellular regulatory volume decrease (RVD) response, though a link between PLM and cell volume regulation has not been studied in the heart. We recently reported a depressed cardiac contractile function in PLM knockout mice in vivo, which was exacerbated in crystalloid-perfused isolated hearts, indicating that these hearts were osmotically challenged. To address this, the present study investigated the role of PLM in osmoregulation in the heart. Isolated PLM wild-type and knockout hearts were perfused with a crystalloid buffer supplemented with mannitol in a bid to prevent perfusate-induced cell swelling and maintain function. Accordingly, and in contrast to wild-type control hearts, contractile function was improved in PLM knockout hearts with 30 mM mannitol. To investigate further, isolated PLM wild-type and knockout cardiomyocytes were subjected to increasing hyposmotic challenges. Initial validation studies showed the IonOptix video edge-detection system to be a simple and accurate 'real-time' method for tracking cell width as a marker of cell size. Myocytes swelled equally in both genotypes, indicating that PLM, when expressed at physiological levels in cardiomyocytes, is not essential to limit water accumulation in response to a hyposmotic challenge. Interestingly, freshly isolated adult cardiomyocytes consistently failed to mount RVDs in response to cell swelling, adding to conflicting reports in the literature. A proposed perturbation of the RVD response as a result of the cell isolation process was not restored, however, with short-term culture in either adult or neonatal cardiomyocytes.

  13. Genetic Determinants of Metabolism and Benign Prostate Enlargement: Associations with Prostate Volume.

    Directory of Open Access Journals (Sweden)

    Ayush Giri

    Full Text Available Prostate enlargement leading to clinical benign prostatic hyperplasia (BPH is associated with metabolic dysregulation and obesity. The genetic basis of this association is unclear. Our objective was to evaluate whether single nucleotide polymorphisms (SNPs previously associated with metabolic disorders are also associated with prostate volume (PV. Participants included 876 men referred for prostate biopsy and found to be prostate cancer free. PV was measured by transrectal ultrasound. Samples were genotyped using the Illumina Cardio-MetaboChip platform. Multivariable adjusted linear regression models were used to evaluate SNPs (additive coding in relation to natural-log transformed (log PV. We compared SNP-PV results from biopsy-negative men to 442 men with low-grade prostate cancer with similar levels of obesity and PV. Beta-coefficients from the discovery and replication samples were then aggregated with fixed effects inverse variance weighted meta-analysis. SNP rs11736129 (near the pseudo-gene LOC100131429 was significantly associated with log-PV (beta: 0.16, p-value 1.16x10(-8 after adjusting for multiple testing. Other noteworthy SNPs that were nominally associated (p-value < 1x10(-4 with log-PV included rs9583484 (intronic SNP in COL4A2, rs10146527 (intronic SNP in NRXN3, rs9909466 (SNP near RPL32P31, and rs2241606 (synonymous SNP in SLC12A7. We found several SNPs in metabolic loci associated with PV. Further studies are needed to confirm our results and elucidate the mechanism between these genetic loci, PV, and clinical BPH.

  14. Genetic Determinants of Metabolism and Benign Prostate Enlargement: Associations with Prostate Volume.

    Science.gov (United States)

    Giri, Ayush; Edwards, Todd L; Motley, Saundra S; Byerly, Susan H; Fowke, Jay H

    2015-01-01

    Prostate enlargement leading to clinical benign prostatic hyperplasia (BPH) is associated with metabolic dysregulation and obesity. The genetic basis of this association is unclear. Our objective was to evaluate whether single nucleotide polymorphisms (SNPs) previously associated with metabolic disorders are also associated with prostate volume (PV). Participants included 876 men referred for prostate biopsy and found to be prostate cancer free. PV was measured by transrectal ultrasound. Samples were genotyped using the Illumina Cardio-MetaboChip platform. Multivariable adjusted linear regression models were used to evaluate SNPs (additive coding) in relation to natural-log transformed (log) PV. We compared SNP-PV results from biopsy-negative men to 442 men with low-grade prostate cancer with similar levels of obesity and PV. Beta-coefficients from the discovery and replication samples were then aggregated with fixed effects inverse variance weighted meta-analysis. SNP rs11736129 (near the pseudo-gene LOC100131429) was significantly associated with log-PV (beta: 0.16, p-value 1.16x10(-8)) after adjusting for multiple testing. Other noteworthy SNPs that were nominally associated (p-value < 1x10(-4)) with log-PV included rs9583484 (intronic SNP in COL4A2), rs10146527 (intronic SNP in NRXN3), rs9909466 (SNP near RPL32P31), and rs2241606 (synonymous SNP in SLC12A7). We found several SNPs in metabolic loci associated with PV. Further studies are needed to confirm our results and elucidate the mechanism between these genetic loci, PV, and clinical BPH.

  15. Early Site Permit Demonstration Program: Guidelines for determining design basis ground motions. Volume 2, Appendices

    Energy Technology Data Exchange (ETDEWEB)

    1993-03-18

    This report develops and applies a methodology for estimating strong earthquake ground motion. The motivation was to develop a much needed tool for use in developing the seismic requirements for structural designs. An earthquake`s ground motion is a function of the earthquake`s magnitude, and the physical properties of the earth through which the seismic waves travel from the earthquake fault to the site of interest. The emphasis of this study is on ground motion estimation in Eastern North America (east of the Rocky Mountains), with particular emphasis on the Eastern United States and southeastern Canada. Eastern North America is a stable continental region, having sparse earthquake activity with rare occurrences of large earthquakes. While large earthquakes are of interest for assessing seismic hazard, little data exists from the region to empirically quantify their effects. The focus of the report is on the attributes of ground motion in Eastern North America that are of interest for the design of facilities such as nuclear power plants. This document, Volume II, contains Appendices 2, 3, 5, 6, and 7 covering the following topics: Eastern North American Empirical Ground Motion Data; Examination of Variance of Seismographic Network Data; Soil Amplification and Vertical-to-Horizontal Ratios from Analysis of Strong Motion Data From Active Tectonic Regions; Revision and Calibration of Ou and Herrmann Method; Generalized Ray Procedure for Modeling Ground Motion Attenuation; Crustal Models for Velocity Regionalization; Depth Distribution Models; Development of Generic Site Effects Model; Validation and Comparison of One-Dimensional Site Response Methodologies; Plots of Amplification Factors; Assessment of Coupling Between Vertical & Horizontal Motions in Nonlinear Site Response Analysis; and Modeling of Dynamic Soil Properties.

  16. Chemically Modified Plastic Tube for High Volume Removal and Collection of Circulating Tumor Cells.

    Directory of Open Access Journals (Sweden)

    Angelo Gaitas

    Full Text Available In this preliminary effort, we use a commercially available and chemically modified tube to selectively capture circulating tumor cells (CTCs from the blood stream by immobilizing human anti-EpCAM antibodies on the tube's interior surface. We describe the requisite and critical steps required to modify a tube into a cancer cell-capturing device. Using these simple modifications, we were able to capture or entrap about 85% of cancer cells from suspension and 44% of cancer cells from spiked whole blood. We also found that the percentage of cells captured was dependent on the tube's length and also the number of cancer cells present. It is our strong belief that with the utilization of appropriate tube lengths and procedures, we can ensure capture and removal of nearly the entire CTC population in whole blood. Importantly after a patient's entire blood volume has circulated through the tube, the tube can then be trypsinized to release the captured live CTCs for further analysis and testing.

  17. Determination of fugitive coal-dust emissions from rotary railcar dumping. Volume 1

    Energy Technology Data Exchange (ETDEWEB)

    Brookman, E.T.; Carnes, D.H.; Catizone, P.A.; Kelley, K.J.

    1984-09-01

    TRC Environmental Consultants, Inc. conducted 72 field tests to determine particulate emission levels for a rotary railcar coal dumper operated by the Potomac Electric Power Company. Approximately 60 parameters were monitored as part of the testing program. These parameters related to particulate emissions, meteorology, and coal properties. The railcar dumper was situated inside of a shed. No other control devices were in operation during the test program. Emissions were measured at the entrance and exit doorways of the shed using the mass flux technique. Background levels were accounted for using the upwind/downwind technique. The amount of particulate material that deposited within the shed was also recorded. The study resulted in the determination of emission levels that were one to two orders of magnitude less than those cited in the literature. These emission levels were found to correlate the strongest with the moisture content of the coal and whether the coal was washed or unwashed.

  18. Combined first pass and equilibrium radionuclide cardiographic determination of stroke volume for quantitation of valvular regurgitation

    DEFF Research Database (Denmark)

    Kelbaek, H; Aldershvile, J; Svendsen, Jesper Hastrup

    1988-01-01

    A new noninvasive procedure for quantitation of cardiac valve regurgitation was evaluated using a combination of first pass and gated equilibrium radionuclide cardiography in 38 subjects with and without cardiac valve disease. Left-sided cardiac catheterization was performed to determine the seve......A new noninvasive procedure for quantitation of cardiac valve regurgitation was evaluated using a combination of first pass and gated equilibrium radionuclide cardiography in 38 subjects with and without cardiac valve disease. Left-sided cardiac catheterization was performed to determine...... with mild mitral incompetence and 2+ aortic regurgitation, 37% in patients with moderate mitral incompetence and 3+ aortic regurgitation and 57% in patients with severe mitral incompetence and 4+ aortic regurgitation. These findings suggest that combined first pass and gated equilibrium radionuclide...

  19. Determination and correlation of mass transfer coefficients in a stirred cell. [Molten Salt Breeder Reactor

    Energy Technology Data Exchange (ETDEWEB)

    Herranz, J.; Bloxom, S.R.; Keeler, J.B.; Roth, S.R.

    1975-12-17

    In the proposed Molten Salt Breeder Reactor flowsheet, a fraction of the rare earth fission products is removed from the fuel salt in mass transfer cells. To obtain design parameters for this extraction, the effect of cell size, blade diameter, phase volume, and agitation rate on the mass transfer for a high density ratio system (mercury/water) in nondispersing square cross section contactors was determined. Aqueous side mass transfer coefficients were measured by polarography over a wide range of operating conditions. Correlations for the experimental mass transfer coefficients as functions of the operating parameters are presented. Several techniques for measuring mercury-side mass transfer coefficients were evaluated and a new one is recommended. (auth)

  20. Mathematical determination of cell population doubling times for multiple cell lines.

    Science.gov (United States)

    Daukste, Liene; Basse, Britta; Baguley, Bruce C; Wall, David J N

    2012-10-01

    Cell cycle times are vital parameters in cancer research, and short cell cycle times are often related to poor survival of cancer patients. A method for experimental estimation of cell cycle times, or doubling times of cultured cancer cell populations, based on addition of paclitaxel (an inhibitor of cell division) has been proposed in literature. We use a mathematical model to investigate relationships between essential parameters of the cell division cycle following inhibition of cell division. The reduction in the number of cells engaged in DNA replication reaches a plateau as the concentration of paclitaxel is increased; this can be determined experimentally. From our model we have derived a plateau log reduction formula for proliferating cells and established that there are linear relationships between the plateau log reduction values and the reciprocal of doubling times (i.e. growth rates of the populations). We have therefore provided theoretical justification of an important experimental technique to determine cell doubling times. Furthermore, we have applied Monte Carlo experiments to justify the suggested linear relationships used to estimate doubling time from 5-day cell culture assays. We show that our results are applicable to cancer cell populations with cell loss present.

  1. Optimization, Application, and Interpretation of Lactate Dehydrogenase Measurements in Microwell Determination of Cell Number and Toxicity

    NARCIS (Netherlands)

    Wolterbeek, H.T.; Van der Meer, A.J.G.M.

    2005-01-01

    The lactate dehydrogenase (LDH) assay was addressed for its sensitivity, disturbances by foaming, and cell number and size. Cells were from a U-251 MG grade IV human glioblastoma brain tumor cell line used in 100-µl well volumes. Cells were counted by microscopy and Coulter counting; assays were LDH

  2. Impedance recordings to determine change in extracellular volume in the brain following cardiac arrest in broiler chickens

    NARCIS (Netherlands)

    Ruis-Heutinck, LFM; Savenije, B; Postema, F; Van Voorst, A; Lambooij, E; Korf, J

    1998-01-01

    The present study describes a method to determine the onset and development of brain damage in broiler chickens. Exsanguination disrupts the brain metabolism and causes the brain to become ischemic. Energy-requiring systems in the cell membrane fail, which results in an ionic shift over the membrane

  3. Galvanic cell without liquid junction for potentiometric determination of copper.

    Science.gov (United States)

    Migdalski, Jan; Błaz, Teresa; Zrałka, Barbara; Lewenstam, Andrzej

    2007-07-01

    This paper describes potentiometric measurements in an integrated galvanic cell with both indicator and reference electrodes. Both electrodes are conducting polymer-based. The copper-sensitive indicator electrode is made by using poly(3,4-ethylenedioxythiophene) (PEDOT) doped with 2-(o-arsenophenylazo)-1,8-dihydroxynaphthalene-3,6-disulphonic sodium salt (Arsenazo-I) as the electroactive substance in the film, while the reference electrode is based on PEDOT doped by 2-morpholineoethanesulfonic acid (MES). It is shown that the galvanic cell can be used for determination of copper both in non-aqueous media (where all PVC-based membranes failed) and in the presence of chloride ions, which disturb the signal of conventional copper ion-selective electrodes with solid-state membranes. It is further shown that the titration of copper ions can be successfully monitored using the described electrochemical cell.

  4. Recurrence pattern of squamous cell carcinoma in the midthoracic esophagus: implications for the clinical target volume design of postoperative radiotherapy

    Directory of Open Access Journals (Sweden)

    Wang X

    2016-10-01

    Full Text Available Xiaoli Wang,1,2,* Yijun Luo,1,2,* Minghuan Li,2 Hongjiang Yan,2 Mingping Sun,2 Tingyong Fan2 1School of Medicine and Life Sciences, Jinan University-Shandong Academy of Medical Sciences, Jinan, Shandong, People’s Republic of China; 2Department of Radiation Oncology, Shandong Cancer Hospital and Institute, Jinan, Shandong, People’s Republic of China *These authors contributed equally to this work Background: Postoperative radiotherapy has shown positive efficacy in lowering the recurrence rate and improving the survival rate for patients with esophageal squamous cell carcinoma (ESCC. However, controversies still exist about the postoperative prophylactic radiation target volume. This study was designed to analyze the patterns of recurrence and to provide a reference for determination of the postoperative radiotherapy target volume for patients with midthoracic ESCC.Patients and methods: A total of 338 patients with recurrent or metastatic midthoracic ESCC after radical surgery were retrospectively examined. The patterns of recurrence including locoregional and distant metastasis were analyzed for these patients.Results: The rates of lymph node (LN metastasis were 28.4% supraclavicular, 77.2% upper mediastinal, 32.0% middle mediastinal, 50.0% lower mediastinal, and 19.5% abdominal LNs. In subgroup analyses, the rate of abdominal LN metastasis was significantly higher in patients with histological node-positive than that in patients with histological node-negative (P=0.033. Further analysis in patients with histological node-positive demonstrated that patients with three or more positive nodes are more prone to abdominal LN metastasis, compared with patients with one or two positive nodes (χ2=4.367, P=0.037. The length of tumor and histological differentiation were also the high-risk factors for abdominal LN metastasis.Conclusion: For midthoracic ESCC with histological node-negative, or one or two positive nodes, the supraclavicular and

  5. Effects of beta interferon on human fibroblasts at different population doubling levels. Proliferation, cell volume, thymidine uptake, and DNA synthesis

    OpenAIRE

    1984-01-01

    Cellular aging had no effect on the ability of beta interferon to increase cell volume and population doubling time in 76-109 cells, a line of human skin fibroblasts. However, DNA synthesis in cells at high population doubling levels (PDL 55-70) was inhibited after 72 h of beta interferon treatment (1,000 U/ml) while no inhibition of DNA synthesis was observed in cells at middle population doubling levels (PDL 30-40).

  6. Applying a new method for direct collection, volume quantification and determination of N2 emission from water.

    Science.gov (United States)

    Liu, Xinhong; Gao, Yan; Wang, Honglian; Guo, Junyao; Yan, Shaohua

    2015-01-01

    The emission of N2 is important to remove excess N from lakes, ponds, and wetlands. To investigate the gas emission from water, Gao et al. (2013) developed a new method using a bubble trap device to collect gas samples from waters. However, the determination accuracy of sampling volume and gas component concentration was still debatable. In this study, the method was optimized for in situ sampling, accurate volume measurement and direct injection to a gas chromatograph for the analysis of N2 and other gases. By the optimized new method, the recovery rate for N2 was 100.28% on average; the mean coefficient of determination (R(2)) was 0.9997; the limit of detection was 0.02%. We further assessed the effects of the new method, bottle full of water, vs. vacuum bag and vacuum vial methods, on variations of N2 concentration as influenced by sample storage times of 1, 2, 3, 5, and 7 days at constant temperature of 15°C, using indices of averaged relative peak area (%) in comparison with the averaged relative peak area of each method at 0 day. The indices of the bottle full of water method were the lowest (99.5%-108.5%) compared to the indices of vacuum bag and vacuum vial methods (119%-217%). Meanwhile, the gas chromatograph determination of other gas components (O2, CH4, and N2O) was also accurate. The new method was an alternative way to investigate N2 released from various kinds of aquatic ecosystems.

  7. Applying a new method for direct collection, volume quantification and determination of N2 emission from water

    Institute of Scientific and Technical Information of China (English)

    Xinhong Liu; Yan Gao; Honglian Wang; Junyao Guo; Shaohua Yan

    2015-01-01

    The emission of N2 is important to remove excess N from lakes,ponds,and wetlands.To investigate the gas emission from water,Gao et al.(2013) developed a new method using a bubble trap device to collect gas samples from waters.However,the determination accuracy of sampling volume and gas component concentration was still debatable.In this study,the method was optimized for in situ sampling,accurate volume measurement and direct injection to a gas chromatograph for the analysis of N2 and other gases.By the optimized new method,the recovery rate for N2 was 100.28% on average; the mean coefficient of determination (R2) was 0.9997; the limit of detection was 0.02%.We further assessed the effects of the new method,bottle full of water,vs.vacuum bag and vacuum vial methods,on variations of N2 concentration as influenced by sample storage times of 1,2,3,5,and 7 days at constant temperature of 15℃,using indices of averaged relative peak area (%) in comparison with the averaged relative peak area of each method at 0 day.The indices of the bottle full of water method were the lowest (99.5%--108.5%) compared to the indices of vacuum bag and vacuum vial methods (119%-217%).Meanwhile,the gas chromatograph determination of other gas components (O2,CH4,and N2O) was also accurate.The new method was an alternative way to investigate N2 released from various kinds of aquatic ecosystems.

  8. EGF stimulates IClswell by a redistribution of proteins involved in cell volume regulation.

    Science.gov (United States)

    Tamma, Grazia; Dossena, Silvia; Nofziger, Charity; Valenti, Giovanna; Svelto, Maria; Paulmichl, Markus

    2011-01-01

    ICln is a multifunctional protein involved in the generation of chloride currents activated during regulatory volume decrease (RVD) after cell swelling (ICl(swell)). Growth factor receptors play a key role in different cellular processes and epidermal growth factor (EGF) regulates swelling-activated chloride permeability. We set out to investigate if the EGF-induced upregulation of ICl(swell) could be explained by a rearrangement of ICln subcellular distribution and interaction with its molecular partners. NIH-3T3 fibroblasts were serum-deprived for 24 hours and stimulated with EGF (40 ng/ml) for 30 minutes. ICl(swell) activation, ICln distribution and interaction with its molecular partner HSPC038 were assessed by whole cell patch clamp and fluorescence resonance energy transfer (FRET). EGF treatment significantly enhanced the direct molecular interaction between ICln and HSPC038 and also resulted in an increase of ICln and HSPC038 association with the plasma membrane. Importantly, these events are associated with a significant increase of ICl(swell). The present data indicate that EGF might exert its role in the modulation of volume-sensitive chloride currents in part through activation and translocation of ICln and HSPC038 to the plasma membrane. Copyright © 2011 S. Karger AG, Basel.

  9. Hemorrhagic shock impairs myocardial cell volume regulation and membrane integrity in dogs

    Energy Technology Data Exchange (ETDEWEB)

    Horton, J.W.

    1987-06-01

    An in vitro myocardial slice technique was used to quantitate alterations in cell volume regulation and membrane integrity after 2 h or hemorrhagic shock. After in vitro incubation in Krebs-Ringer-phosphate medium containing trace (/sup 14/C)inulin, values (ml H/sub 2/O/g dry wt) for control nonshocked myocardial slices were 4.03 /plus minus/ 0.11 (SE) for total water, 2.16 /plus minus/ 0.07 for inulin impermeable space, and 1.76 /plus minus/ 0.15 for inulin diffusible space. Shocked myocardial slices showed impaired response to cold incubation. After 2 h of in vivo shock, total tissue water, inulin diffusible space, and inulin impermeable space increased significantly for subendocardium, whereas changes in subepicardium parameters were minimal. Shock-induced cellular swelling was accompanied by an increased total tissue sodium, but no change in tissue potassium. Calcium entry blockade in vivo significantly reduced subendocardial total tissue water as compared with shock-untreated dogs. In addition, calcium entry blockade reduced shock-induced increases in inulin diffusible space. In vitro myocardial slice studies confirm alterations in subendocardial membrane integrity after 2 h of in vivo hemorrhagic shock. Shock-induced abnormalities in myocardial cell volume regulation are reduced by calcium entry blockade in vivo.

  10. Cell and Tissue Organization in the Circulatory and Ventilatory Systems Volume 1 Signaling in Cell Organization, Fate, and Activity, Part A Cell Structure and Environment

    CERN Document Server

    Thiriet, Marc

    2011-01-01

    The volumes in this authoritative series present a multidisciplinary approach to modeling and simulation of flows in the cardiovascular and ventilatory systems, especially multiscale modeling and coupled simulations. The cardiovascular and respiratory systems are tightly coupled, as their primary function is to supply oxygen to and remove carbon dioxide from the body's cells. Because physiological conduits have deformable and reactive walls, macroscopic flow behavior and prediction must be coupled to nano- and microscopic events in a corrector scheme of regulated mechanisms. Therefore, investigation of flows of blood and air in physiological conduits requires an understanding of the biology, chemistry, and physics of these systems together with the mathematical tools to describe their functioning.  The present volume is devoted to cellular events that allow adaptation to environmental conditions, particularly mechanotransduction. It begins with cell organization and a survey of cell types in the vasculatur...

  11. Effects of pressure- or volume-overload hypertrophy on passive stiffness in isolated adult cardiac muscle cells

    Science.gov (United States)

    Kato, S.; Koide, M.; Cooper, G. 4th; Zile, M. R.

    1996-01-01

    It has been hypothesized that the changes in myocardial stiffness induced by chronic hemodynamic overloading are dependent on changes in the passive stiffness of the cardiac muscle cell (cardiocyte). However, no previous studies have examined the passive constitutive properties of cardiocytes isolated from animals with myocardial hypertrophy. Accordingly, changes in relative passive stiffness of cardiocytes isolated from animals with chronic pressure- or volume-overload hypertrophy were determined by examining the effects of anisosmotic stress on cardiocyte size. Anisosmotic stress was produced by altering superfusate osmolarity. Hypertrophied cardiocytes were enzymatically isolated from 16 adult cats with right ventricular (RV) pressure-overload hypertrophy induced by pulmonary artery banding (PAB) and from 6 adult cats with RV volume-overload hypertrophy induced by creating an atrial septal defect (ASD). Left ventricular (LV) cardiocytes from each cat served as nonhypertrophied, normally loaded, same-animal controls. Superfusate osmolarity was decreased from 305 +/- 3 to 135 +/- 5 mosM and increased to 645 +/- 4 mosM. During anisosmotic stress, there were no significant differences between hypertrophied RV and normal LV cardiocytes in pressure overload PAB cats with respect to percent change in cardiocyte area (47 +/- 2% in RV vs. 48 +/- 2% in LV), diameter (46 +/- 3% in RV vs. 48 +/- 2% in LV), or length (2.4 +/- 0.2% in RV vs. 2.0 +/- 0.3% in LV), or sarcomere length (1.5 +/- 0.1% in RV vs. 1.3 +/- 0.3% in LV). Likewise, there were no significant differences in cardiocyte strain between hypertrophied RV and normal LV cardiocytes from ASD cats. In conclusion, chronic pressure-overload hypertrophy and chronic volume-overload hypertrophy did not alter the cardiocyte response to anisosmotic stress. Thus chronic overload hypertrophy did not alter relative passive cardiocyte stiffness.

  12. Effects of pressure- or volume-overload hypertrophy on passive stiffness in isolated adult cardiac muscle cells

    Science.gov (United States)

    Kato, S.; Koide, M.; Cooper, G. 4th; Zile, M. R.

    1996-01-01

    It has been hypothesized that the changes in myocardial stiffness induced by chronic hemodynamic overloading are dependent on changes in the passive stiffness of the cardiac muscle cell (cardiocyte). However, no previous studies have examined the passive constitutive properties of cardiocytes isolated from animals with myocardial hypertrophy. Accordingly, changes in relative passive stiffness of cardiocytes isolated from animals with chronic pressure- or volume-overload hypertrophy were determined by examining the effects of anisosmotic stress on cardiocyte size. Anisosmotic stress was produced by altering superfusate osmolarity. Hypertrophied cardiocytes were enzymatically isolated from 16 adult cats with right ventricular (RV) pressure-overload hypertrophy induced by pulmonary artery banding (PAB) and from 6 adult cats with RV volume-overload hypertrophy induced by creating an atrial septal defect (ASD). Left ventricular (LV) cardiocytes from each cat served as nonhypertrophied, normally loaded, same-animal controls. Superfusate osmolarity was decreased from 305 +/- 3 to 135 +/- 5 mosM and increased to 645 +/- 4 mosM. During anisosmotic stress, there were no significant differences between hypertrophied RV and normal LV cardiocytes in pressure overload PAB cats with respect to percent change in cardiocyte area (47 +/- 2% in RV vs. 48 +/- 2% in LV), diameter (46 +/- 3% in RV vs. 48 +/- 2% in LV), or length (2.4 +/- 0.2% in RV vs. 2.0 +/- 0.3% in LV), or sarcomere length (1.5 +/- 0.1% in RV vs. 1.3 +/- 0.3% in LV). Likewise, there were no significant differences in cardiocyte strain between hypertrophied RV and normal LV cardiocytes from ASD cats. In conclusion, chronic pressure-overload hypertrophy and chronic volume-overload hypertrophy did not alter the cardiocyte response to anisosmotic stress. Thus chronic overload hypertrophy did not alter relative passive cardiocyte stiffness.

  13. Determination of the Representative Elementary Volume for the study of sandstones and siltstones by X-Ray microtomography

    Directory of Open Access Journals (Sweden)

    Jaquiel Salvi Fernandes

    2012-08-01

    Full Text Available X-Ray computerized microtomography (µ-CT besides providing two-dimensional images (2-D of the transversal sections of the sample, the biggest attraction of the methodology is the rendering of three-dimensional images (3-D, enabling a more real analysis of the porous structure of the rock. However, the reconstruction, visualization and analysis of such 3-D images are limited in computer terms. Thus, it is not always possible to reconstruct the images with the total size of the microtomographed sample. Therefore, this study aims at determining the Representative Elementary Volume (REV in reservoir rocks concerning their porosity. In order to collect microtomographic data from reservoir rocks, a microtomograph Skyscan model 1172 was utilized for the sandstone and siltstone samples scanning. After the analysis of the graphs obtained by REV, it was concluded that the most adequate dimensions for the reconstructed volume in each analyzed sample were approximately 1400 × 1400 × 1400 µm, which are dimensions that can easily be reconstructed, visualized and analyzed.

  14. Pancreatic islet amyloidosis, β-cell apoptosis, and α-cell proliferation are determinants of islet remodeling in type-2 diabetic baboons

    Science.gov (United States)

    Guardado-Mendoza, Rodolfo; Davalli, Alberto M.; Chavez, Alberto O.; Hubbard, Gene B.; Dick, Edward J.; Majluf-Cruz, Abraham; Tene-Perez, Carlos E.; Goldschmidt, Lukasz; Hart, John; Perego, Carla; Comuzzie, Anthony G.; Tejero, Maria Elizabeth; Finzi, Giovanna; Placidi, Claudia; La Rosa, Stefano; Capella, Carlo; Halff, Glenn; Gastaldelli, Amalia; DeFronzo, Ralph A.; Folli, Franco

    2009-01-01

    β-Cell dysfunction is an important factor in the development of hyperglycemia of type-2 diabetes mellitus, and pancreatic islet amyloidosis (IA) has been postulated to be one of the main contributors to impaired insulin secretion. The aim of this study was to evaluate the correlation of IA with metabolic parameters and its effect on islets of Langerhans remodeling and relative endocrine-cell volume in baboons. We sequenced the amylin peptide, determined the fibrillogenic propensities, and evaluated pancreatic histology, clinical and biochemical characteristics, and endocrine cell proliferation and apoptosis in 150 baboons with different metabolic status. Amylin sequence in the baboon was 92% similar to humans and showed superimposable fibrillogenic propensities. IA severity correlated with fasting plasma glucose (FPG) (r = 0.662, P < 0.001) and HbA1c (r = 0.726, P < 0.001), as well as with free fatty acid, glucagon values, decreased homeostasis model assessment (HOMA) insulin resistance, and HOMA-B. IA severity was associated with a decreased relative β-cell volume, and increased relative α-cell volume and hyperglucagonemia. These results strongly support the concept that IA and β-cell apoptosis in concert with α-cell proliferation and hypertrophy are key determinants of islets of Langerhans “dysfunctional remodeling” and hyperglycemia in the baboon, a nonhuman primate model of type-2 diabetes mellitus. The most important determinants of IA were age and FPG (R2 = 0.519, P < 0.0001), and different FPG levels were sensitive and specific to predict IA severity. Finally, a predictive model for islet amyloid severity was generated with age and FPG as required variables. PMID:19666551

  15. Determinants of intrinsic radiosensitivity of mammalian cell lines

    Energy Technology Data Exchange (ETDEWEB)

    Radford, I.R. [Peter MacCallum Cancer Institute, East Melbourne, VIC (Australia). Research Division

    1998-12-31

    Differences in the radiosensitivity of normal and cancerous cells could arise in various ways. Although there is no compelling data to support the view, the currently prevailing opinion is that differences in radiosensitivity are related to differences in some aspect of enzymatic DNA repair. A test of the importance of possible differences in enzymatic DNA repair in determining relative radiosensitivity would be to compare lethality in cells containing equivalent numbers of DNA lesions. Six cell lines were used in these studies: two Chinese hamster (CHO and V79) and a monkey (Vero) fibroblast-like line, a mouse melanoma line (B16-F1), and a rat (RUC-2) and a human (SQ-20B) carcinoma line. This group of cell lines displays a wide range of sensitivities to external beam low-LET radiation, ranging from the relatively radiosensitive B16-F1 and Vero lines through to the highly radioresistant RUC-2 line. However, it is important to note that none of the lines has a demonstrated defect in enzymatic DNA repair and that all appear to die by necrosis following a lethal radiation insult. Despite having significantly different radiosensitivities, CHO and V79 cells showed comparable responses to DNA-associated {sup 125}I-decays with D{sub o} values of around 65. More surprisingly, the radiosensitive B16-F1 line and the radioresistant RUC-2 line both had responses with D{sub o} values of around 133 {sup 125}I-decays. The factor of two difference between the D{sub o} values for these two pairs of cell lines is probably attributable to CHO and V79 cells being pseudo-diploid whereas B 16-F1 and RUC2 appear to have derived from tetraploid cells. The generality of the above result, for DNA lesions of different quality, was tested by comparing the sensitivities of CHO and V79 cells to DNA-associated {sup 3}H-decays. Again, consistent with the {sup 125}I-decay data, there was no significant difference in the D{sub o} values for these lines. Our {sup 3}H- and {sup 125}I-decay data are

  16. Stem cell harvesting protocol research in autologous transplantation setting: Large volume vs. conventional cytapheresis

    Directory of Open Access Journals (Sweden)

    Balint Bela

    2008-01-01

    Full Text Available Background/Aim. The use of peripheral blood as a source of hematopoietic stem cells (SCs is progressively increasing and has nearly supplanted bone marrow transplantation. Interpatient variability in the degree and kinetics of SC mobilization into peripheral blood is an expected event after conventional chemotherapy-based treatment, followed by sequential administration of recombinant granulocyte-colony- stimulating factor (rHu-CSF. In this study, specific factors associated with the application of two different SC-harvesting approaches, including the use of large volume leukapheresis (LVL vs. repetitive conventional apheresis (RCA, were analyzed. The basic goal of the study was to evaluate the influence of apheresis protocol (collection timing, processed blood volume and cell yield upon the clinical outcome of transplantation. Methods. Results obtained by LVL (76 pts and RCA (20 pts - control group were compared. The SC mobilizing regimen used was cyclophosphamide (4-7 g/m2 or polychemotherapy and rHuG-CSF 10-16 μg/kg of body mess (bm per day. Cell harvesting was performed using COBE-Spectra (Caridian-BCT, USA. The volume of processed blood in LVL setting was ≥ 3.5 - fold of the patient's circulating blood quantity (ranged from 12.7 to 37.8 l. All patients tolerated well the use of intensive treatment, without any side or adverse effects. Our original controlled-rate cryopreservation was carried out with 10% dimethyl sulfoxide (DMSO using Planer R203/200R or Planer 560-16 equipments (Planer Products Ltd, UK. Total nucleated cell (NC and mononuclear cell (MNC counts were examined by flow cytometry (Advia-2120 Bayer, Germany; Technicon H-3 System, USA. The CD34+ cell surface antigen was investigated by the EPICS XL-MCL device (Coulter, Germany. Results. Performing LVL-apheresis, high-level MNC and CD34+ cell yields (7.6±4.6 × 108/kg bm and 11.8±6.5 × 106/kg bm, respectively were obtained. As a result, rapid hematopoietic reconstitution

  17. Determination of telomerase activity in stem cells and non-stem cells of breast cancer

    Institute of Scientific and Technical Information of China (English)

    LI Zhi; HE Yanli; ZHANG Jiahua; ZHANG Jinghui; HUANG Tao

    2007-01-01

    Although all normal tissue cells,including stem cells,are genetically homologous,variation in gene expression patterns has already determined the distinct roles for individual cells in the physiological process due to the occurrence of epigenetic modification.This is of special importance for the existenee of tissue stem cells because they are exclusively immortal within the body,capable of selfreplicating and differentiating by which tissues renew and repair itself and the total tissue cell population maintains a steady-state.Impairment of tissue stem cells is usually accompanied by a reduction in cell number,slows down the repair process and causes hypofunction.For instance,chemotherapy usually leads to depression of bone marrow and hair loss.Cellular aging is closely associated with the continuous erosion of the telomere while activation of telomerase repairs and maintains telomeres,thus slowing the aging process and prolonging cell life.In normal adults,telomerase activation mainly presents in tissue stem cells and progenitor cells giving them unlimited growth potential.Despite the extensive demonstration of telomerase activation in malignancy(>80%),scientists found that heterogeneity also exists among the tumor cells and only minorities of cells,designated as cancer stem cells,andergo processes analogous to the self-renewal and differentiation of normal stem ceils while the rest have limited lifespans.In this study,telomerase activity was measured and compared in breast cancer stem cells and non-stem cells that were phenotypically sorted by examining surface marker expression.The results indicated that cancer stem cells show a higher level of enzyme activity than non-stem cells.In addition,associated with the repair of cancer tissue(or relapse)after chemotherapy,telomerase activity in stem cells was markedly increased.

  18. Determining the benefits of Vorticella cell body motion

    Science.gov (United States)

    Specht, Matty C.; Pepper, Rachel E.

    2016-11-01

    Microscopic sessile suspension feeders are single-celled organisms found in aquatic ecosystems. They live attached to underwater surfaces and create a fluid flow in order to feed on bacteria and debris. They participate in the natural degradation of contaminants in water. Understanding the fluid flow they create enhances our knowledge of their environmental impact. One type of suspension feeder, Vorticella, have been observed to vary their cell body orientation with respect to their surface, but the benefits of this motion are still unknown. We use simulations to investigate the effect of Vorticella body motion on the feeding current and the nutrient flux to the cell body to determine whether or not the motion increases nutrient consumption. We determine the nutrient flux using COMSOL Multiphysics software to solve the advection-diffusion equation with the flow given by a stokeslet model. We use a range of motions similar and dissimilar to that of live Vorticella. We find that most patterns of motion do not increase the nutrient flux, since the Vorticella feed from regions where they already have depleted the water of nutrients. However, it is possible that their motion could help the Vorticella find nutrients that are inhomogenously distributed in water.

  19. Two-dimensional echocardiographic determination of left atrial emptying volume: a noninvasive index in quantifying the degree of nonrheumatic mitral regurgitation.

    Science.gov (United States)

    Ren, J F; Kotler, M N; DePace, N L; Mintz, G S; Kimbiris, D; Kalman, P; Ross, J

    1983-10-01

    Several noninvasive techniques, including radionuclide angiography and Doppler echocardiography, have attempted to measure the regurgitant volume in patients with mitral regurgitation; however, none of these techniques are entirely satisfactory. Utilizing a computerized light pen method for tracing the left atrial endocardial border during systole and diastole in two orthogonal planes (apical four and two chamber views), biplane volume determinations were calculated in 12 normal subjects and 30 patients with nonrheumatic mitral regurgitation. Left atrial emptying volume determinations were performed by subtracting the left atrial end-diastolic volume from the left atrial end-systolic volume. The degree of mitral regurgitation was visually assessed as normal (0, trivial, Group I, 12 patients), mild (1+, Group II, 4 patients), moderate (2+, Group III, 8 patients), moderately severe (3+, Group IV, 12 patients) and severe (4+, Group V, 6 patients) by contrast left ventricular angiography and also quantitatively by regurgitant fraction at cardiac catheterization. All 18 patients with moderately severe (Group IV) and severe (Group V) mitral regurgitation had a left atrial emptying volume greater than 40 ml compared with none of the normal subjects and patients with mild (Group II) or moderate (Group III) mitral regurgitation. There was good correlation between left atrial emptying volume and mitral regurgitant fraction (r = 0.85, p less than 0.01). Thus, in patients with nonrheumatic mitral regurgitation, left atrial emptying volume is useful in separating mild from severe mitral regurgitation.

  20. Determination of amphetamines in hair by GC/MS after small-volume liquid extraction and microwave derivatization.

    Science.gov (United States)

    Meng, Pinjia; Zhu, Dan; He, Hongyuan; Wang, Yanyan; Guo, Fei; Zhang, Liang

    2009-09-01

    We report here on the results of a procedure for the determination of amphetamine drugs in hair. The procedure is simple and sensitive. The results from the procedure using small-volume extraction matches perfectly with those either from using the derivatization method or selected ion monitoring (SIM) detection. We validated our method using four different amine drugs, including amphetamine, methamphetamine, methylenedioxy-amphetamine and methylenedioxy-methamphetamine. The detection limit for these drugs is about 50 +/- 7.5 pg/mg in hair and the intra-day and inter-day reproducibility are within 15% at most drug concentrations. Moreover, we also showed the utility of the procedure in analyses of authentic hair samples taken from amphetamine abusers, and demonstrated that the method meets the requirement for the analysis of a trace amounts of amphetamines in human hair.

  1. Determining the unsaturated hydraulic conductivity of a compacted sand-bentonite mixture under constant volume and free-swell conditions

    CERN Document Server

    Cui, Yu-Jun; Loiseau, Cyril; Delage, Pierre; 10.1016/j.pce.2008.10.017

    2008-01-01

    Highly compacted sand-bentonite mixtures are often considered as possible engineered barriers in deep high-level radioactive waste disposals. In-situ, the saturation of these barriers from their initially unsaturated state is a complex hydro-mechanical coupled process in which temperature effects also play a role. The key parameter of this process is the unsaturated hydraulic conductivity of the barrier. In this paper, isothermal infiltration experiments were conducted to determine the unsaturated hydraulic conductivity according to the instantaneous profile method. To do so, total suction changes were monitored at different locations along the soil specimen by using resistivity relative humidity probes. Three constant volume infiltration tests were conducted showing, unexpectedly, a decrease of the hydraulic conductivity during infiltration. One test performed under free-swell conditions showed the opposite and standard trend. These observations were interpreted in terms of microstructure changes during wett...

  2. Glucose Levels in Culture Medium Determine Cell Death Mode in MPP(+)-treated Dopaminergic Neuronal Cells.

    Science.gov (United States)

    Yoon, So-Young; Oh, Young J

    2015-09-01

    We previously demonstrated that 1-methyl-4-phenylpyridinium (MPP(+)) causes caspase-independent, non-apoptotic death of dopaminergic (DA) neuronal cells. Here, we specifically examined whether change of glucose concentration in culture medium may play a role for determining cell death modes of DA neurons following MPP(+) treatment. By incubating MN9D cells in medium containing varying concentrations of glucose (5~35 mM), we found that cells underwent a distinct cell death as determined by morphological and biochemical criteria. At 5~10 mM glucose concentration (low glucose levels), MPP(+) induced typical of the apoptotic dell death accompanied with caspase activation and DNA fragmentation as well as cell shrinkage. In contrast, MN9D cells cultivated in medium containing more than 17.5 mM (high glucose levels) did not demonstrate any of these changes. Subsequently, we observed that MPP(+) at low glucose levels but not high glucose levels led to ROS generation and subsequent JNK activation. Therefore, MPP(+)-induced cell death only at low glucose levels was significantly ameliorated following co-treatment with ROS scavenger, caspase inhibitor or JNK inhibitor. We basically confirmed the quite similar pattern of cell death in primary cultures of DA neurons. Taken together, our results suggest that a biochemically distinct cell death mode is recruited by MPP(+) depending on extracellular glucose levels.

  3. The cell-cycle state of stem cells determines cell fate propensity.

    Science.gov (United States)

    Pauklin, Siim; Vallier, Ludovic

    2013-09-26

    Self-renewal and differentiation of stem cells are fundamentally associated with cell-cycle progression to enable tissue specification, organ homeostasis, and potentially tumorigenesis. However, technical challenges have impaired the study of the molecular interactions coordinating cell fate choice and cell-cycle progression. Here, we bypass these limitations by using the FUCCI reporter system in human pluripotent stem cells and show that their capacity of differentiation varies during the progression of their cell cycle. These mechanisms are governed by the cell-cycle regulators cyclin D1-3 that control differentiation signals such as the TGF-β-Smad2/3 pathway. Conversely, cell-cycle manipulation using a small molecule directs differentiation of hPSCs and provides an approach to generate cell types with a clinical interest. Our results demonstrate that cell fate decisions are tightly associated with the cell-cycle machinery and reveal insights in the mechanisms synchronizing differentiation and proliferation in developing tissues.

  4. The roles and regulation of Sertoli cells in fate determinations of spermatogonial stem cells and spermatogenesis.

    Science.gov (United States)

    Hai, Yanan; Hou, Jingmei; Liu, Yun; Liu, Yang; Yang, Hao; Li, Zheng; He, Zuping

    2014-05-01

    Spermatogenesis is a complex process by which spermatogonial stem cells (SSCs) self-renew and differentiate into spermatozoa under the elaborate coordination of testicular microenvironment, namely, niche. Sertoli cells, which locate around male germ cells, are the most critical component of the niche. Significant progress has recently been made by peers and us on uncovering the effects of Sertoli cells on regulating fate determinations of SSCs. Here we addressed the roles and regulation of Sertoli cells in normal and abnormal spermatogenesis. Specifically, we summarized the biological characteristics of Sertoli cells, and we emphasized the roles of Sertoli cells in mediating the self-renewal, differentiation, apoptosis, de-differentiation, and trans-differentiation of SSCs. The association between abnormal function of Sertoli cells and impaired spermatogenesis was discussed. Finally, we highlighted several issues to be addressed for further investigation on the effects and mechanisms of Sertoli cells in spermatogenesis. Since Sertoli cells are the key supportive cells for SSCs and they are very receptive to modification, a better understanding of the roles and regulation of Sertoli cells in SSC biology and spermatogenesis would make it feasible to identify novel targets for gene therapy of male infertility as well as seek more efficient and safer strategies for male contraception.

  5. Comparison between methods using copper, lanthanum, and colorimetry for the determination of the cation exchange capacity of plant cell walls.

    Science.gov (United States)

    Wehr, J Bernhard; Blamey, F Pax C; Menzies, Neal W

    2010-04-28

    The determination of the cation exchange capacity (CEC) of plant cell walls is important for many physiological studies. We describe the determination of cell wall CEC by cation binding, using either copper (Cu) or lanthanum (La) ions, and by colorimetry. Both cations are strongly bound by cell walls, permitting fast and reproducible determinations of the CEC of small samples. However, the dye binding methods using two cationic dyes, Methylene Blue and Toluidine Blue, overestimated the CEC several-fold. Column and centrifugation methods are proposed for CEC determination by Cu or La binding; both provide similar results. The column method involves packing plant material (2-10 mg dry mass) in a chromatography column (10 mL) and percolating with 20 bed volumes of 1 mM La or Cu solution, followed by washing with deionized water. The centrifugation method uses a suspension of plant material (1-2 mL) that is centrifuged, and the pellet is mixed three times with 10 pellet volumes of 1 mM La or Cu solution followed by centrifugation and final washing with deionized water. In both methods the amount of La or Cu bound to the material was determined by spectroscopic methods.

  6. Potassium-chloride cotransporter 3 interacts with Vav2 to synchronize the cell volume decrease response with cell protrusion dynamics.

    Directory of Open Access Journals (Sweden)

    Adèle Salin-Cantegrel

    Full Text Available Loss-of-function of the potassium-chloride cotransporter 3 (KCC3 causes hereditary motor and sensory neuropathy with agenesis of the corpus callosum (HMSN/ACC, a severe neurodegenerative disease associated with defective midline crossing of commissural axons in the brain. Conversely, KCC3 over-expression in breast, ovarian and cervical cancer is associated with enhanced tumor cell malignancy and invasiveness. We identified a highly conserved proline-rich sequence within the C-terminus of the cotransporter which when mutated leads to loss of the KCC3-dependent regulatory volume decrease (RVD response in Xenopus Laevis oocytes. Using SH3 domain arrays, we found that this poly-proline motif is a binding site for SH3-domain containing proteins in vitro. This approach identified the guanine nucleotide exchange factor (GEF Vav2 as a candidate partner for KCC3. KCC3/Vav2 physical interaction was confirmed using GST-pull down assays and immuno-based experiments. In cultured cervical cancer cells, KCC3 co-localized with the active form of Vav2 in swelling-induced actin-rich protruding sites and within lamellipodia of spreading and migrating cells. These data provide evidence of a molecular and functional link between the potassium-chloride co-transporters and the Rho GTPase-dependent actin remodeling machinery in RVD, cell spreading and cell protrusion dynamics, thus providing new insights into KCC3's involvement in cancer cell malignancy and in corpus callosum agenesis in HMSN/ACC.

  7. Variation of Red Blood Cell Distribution Width and Mean Platelet Volume after Moderate Endurance Exercise

    Directory of Open Access Journals (Sweden)

    Giuseppe Lippi

    2014-01-01

    Full Text Available Although physical exercise strongly influences several laboratory parameters, data about the hematological changes after medium distance running are scarce. We studied 31 middle-trained athletes (mean training regimen 217±32 min/week who performed a 21.1 km, half-marathon run. Blood samples were collected before the run, at the end, and 3 and 20 hours thereafter. The complete blood count was performed on Advia 2120 and included red blood cell (RBC, reticulocyte, and platelet counts; hemoglobin; mean corpuscular volume (MCV; mean corpuscular hemoglobin (MCH; reticulocyte haemoglobin content (Ret CHR; RBC distribution width (RDW, mean platelet volume (MPV. No significant variations were observed for MCH and Ret CHR. The RBC, reticulocyte, and hemoglobin values modestly decreased after the run. The MCV significantly increased at the end of running but returned to baseline 3 hours thereafter. The RDW constantly increased, reaching a peak 20 hours after the run. The platelet count and MPV both increased after the run and returned to baseline 3 hours thereafter. These results may have implications for definition of reference ranges and antidoping testing, and may also contribute to explaining the relationship between endurance exercise and mortality, since previous studies reported that RDW and MPV may be significantly associated with cardiovascular disease.

  8. Cell volume changes affect gluconeogenesis in the perfused liver of the catfish Clarias batrachus

    Indian Academy of Sciences (India)

    Carina Goswami; Shritapa Datta; Kuheli Biswas; Nirmalendu Saha

    2004-09-01

    In addition to lactate and pyruvate, some amino acids were found to serve as potential gluconeogenic substrates in the perfused liver of Clarias batrachus. Glutamate was found to be the most effective substrate, followed by lactate, pyruvate, serine, ornithine, proline, glutamine, glycine, and aspartate. Four gluconeogenic enzymes, namely phosphoenolpyruvate carboxykinase (PEPCK), pyruvate carboxylase (PC), fructose 1,6-bisphosphatase (FBPase) and glucose 6-phosphatase (G6Pase) could be detected mainly in liver and kidney, suggesting that the latter are the two major organs responsible for gluconeogenic activity in this fish. Hypo-osmotically induced cell swelling caused a significant decrease of gluconeogenic efflux accompanied with significant decrease of activities of PEPCK, FBPase and G6Pase enzymes in the perfused liver. Opposing effects were seen in response to hyper-osmotically induced cell shrinkage. These changes were partly blocked in the presence of cycloheximide, suggesting that the aniso-osmotic regulations of gluconeogenesis possibly occurs through an inverse regulation of enzyme proteins and/or a regulatory protein synthesis in this catfish. In conclusion, gluconeogenesis appears to play a vital role in C. batrachus in maintaining glucose homeostasis, which is influenced by cell volume changes possibly for proper energy supply under osmotic stress.

  9. Vertebrate blood cell volume increases with temperature: implications for aerobic activity

    Directory of Open Access Journals (Sweden)

    James F. Gillooly

    2014-04-01

    Full Text Available Aerobic activity levels increase with body temperature across vertebrates. Differences in these levels, from highly active to sedentary, are reflected in their ecology and behavior. Yet, the changes in the cardiovascular system that allow for greater oxygen supply at higher temperatures, and thus greater aerobic activity, remain unclear. Here we show that the total volume of red blood cells in the body increases exponentially with temperature across vertebrates, after controlling for effects of body size and taxonomy. These changes are accompanied by increases in relative heart mass, an indicator of aerobic activity. The results point to one way vertebrates may increase oxygen supply to meet the demands of greater activity at higher temperatures.

  10. Determination of the minimum size of a statistical representative volume element from a fibre-reinforced composite based on point pattern statistics

    DEFF Research Database (Denmark)

    Hansen, Jens Zangenberg; Brøndsted, Povl

    2013-01-01

    In a previous study, Trias et al. [1] determined the minimum size of a statistical representative volume element (SRVE) of a unidirectional fibre-reinforced composite primarily based on numerical analyses of the stress/strain field. In continuation of this, the present study determines the minimum...

  11. Glucose concentration and medium volume influence cell viability and glycosaminoglycan synthesis in chondrocyte-seeded alginate constructs.

    Science.gov (United States)

    Heywood, Hannah K; Bader, Dan L; Lee, David A

    2006-12-01

    Increasing the thickness of tissue-engineered cartilage is associated with loss of chondrocyte viability and biosynthetic activity at the tissue center. Exceptionally high volumes of culture medium, however, can maintain cellularity and glycosaminoglycan synthesis throughout 4-mm-thick constructs. We hypothesized that glucose supplementation could replicate the augmentation of tissue formation achieved by medium volume. Chondrocyte-alginate constructs (40x10(6) cells/mL) were cultured for 14 days in 0.4-6.4 mL/10(-6) cells of either low- (5.1 mM) or high- (20.4 mM) glucose medium. Glucose was critical to chondrocyte viability, and glucose uptake increased significantly (P cells of low-glucose medium had a mass of 172 +/- 6.1 mg and glycosaminoglycan (GAG) content of 0.32 +/- 0.03 mg (mean +/- standard deviation). A 4-fold increase in medium volume increased the final construct mass by 44% and GAG content by 207%. An equivalent increase in glucose supply in the absence of volume change increased these parameters by just 10% and 73%, respectively. A similar trend was observed from 0.8 to 3.2 mL/10(-6) cells, when maximal values of construct GAG content and mass were obtained. Therefore, medium volume remains an important consideration for the optimal culture of tissue-engineered cartilage.

  12. Electrolyte Volume Effects on Electrochemical Performance and Solid Electrolyte Interphase in Si-Graphite/NMC Lithium-Ion Pouch Cells

    Energy Technology Data Exchange (ETDEWEB)

    An, Seong Jin; Li, Jianlin; Daniel, Claus; Meyer, Harry M.; Trask, Stephen E.; Polzin, Bryant J.; Wood, David L.

    2017-05-23

    This study aims to explore the correlations between electrolyte volume, electrochemical performance, and properties of the solid electrolyte interphase in pouch cells with Si-graphite composite anodes. The electrolyte is 1.2 M LiPF6 in ethylene carbonate:ethylmethyl carbonate with 10 wt.% fluoroethylene carbonate. Single layer pouch cells (100 mAh) were constructed with 15 wt.% Si-graphite / LiNi0.5Mn0.3CO0.2O2 electrodes. It is found that a minimum electrolyte volume factor of 3.1 times the total pore volume of cell components (cathode, anode, and separator) is needed for better cycling stability. Less electrolyte causes increases in ohmic and charge transfer resistances. Lithium dendrites are observed when the electrolyte volume factor is low. The resistances from the anodes become significant as the cells are discharged. Solid electrolyte interphase thickness grows as the electrolyte volume factor increases and is non-uniform after cycling.

  13. Progenitor/Stem Cell Fate Determination: Interactive Dynamics of Cell Cycle and Microvesicles

    Science.gov (United States)

    Lee, David; Puente, Napoleon; Faradyan, Sam; Sears, Edmund H.; Amaral, Ashley; Goldberg, Laura; Dooner, Mark S.; Pereira, Mandy; Quesenberry, Peter J.

    2012-01-01

    We have shown that hematopoietic stem/progenitor cell phenotype and differentiative potential change throughout cell cycle. Lung-derived microvesicles (LDMVs) also change marrow cell phenotype by inducing them to express pulmonary epithelial cell-specific mRNA and protein. These changes are accentuated when microvesicles isolated from injured lung. We wish to determine if microvesicle-treated stem/progenitor cell phenotype is linked to cell cycle and to the injury status of the lung providing microvesicles. Lineage depleted, Sca-1+ (Lin-/Sca-1+) marrow isolated from mice were cultured with interleukin 3 (IL-3), IL-6, IL-11, and stem cell factor (cytokine-cultured cells), removed at hours zero (cell cycle phase G0/G1), 24 (late G1/early S), and 48 (late S/early G2/M), and cocultured with lung tissue, lung conditioned media (LCM), or LDMV from irradiated or nonirradiated mice. Alternatively, Lin-/Sca-1+ cells not exposed to exogenous cytokines were separated into G0/G1 and S/G2/M cell cycle phase populations by fluorescence-activated cell sorting (FACS) and used in coculture. Separately, LDMV from irradiated and nonirradiated mice were analyzed for the presence of adhesion proteins. Peak pulmonary epithelial cell-specific mRNA expression was seen in G0/G1 cytokine-cultured cells cocultured with irradiated lung and in late G1/early S cells cocultured with nonirradiated lung. The same pattern was seen in cytokine-cultured Lin-/Sca-1 cells cocultured with LCM and LDMV and when FACS-separated Lin-/Sca-1 cells unexposed to exogenous cytokines were used in coculture. Cells and LDMV expressed adhesion proteins whose levels differed based on cycle status (cells) or radiation injury (LDMV), suggesting a mechanism for microvesicle entry. These data demonstrate that microvesicle modification of progenitor/stem cells is influenced by cell cycle and the treatment of the originator lung tissue. PMID:22214238

  14. Mean platelet volume and red cell distribution width levels in initial evaluation of panic disorder

    Directory of Open Access Journals (Sweden)

    Asoglu M

    2016-09-01

    Full Text Available Mehmet Asoglu,1 Mehmet Aslan,2 Okan Imre,1 Yuksel Kivrak,3 Oznur Akil,1 Emin Savik,4 Hasan Buyukaslan,5 Ulker Fedai,1 Abdurrahman Altındag6 1Department of Psychiatry, Faculty of Medicine, Harran University, Sanliurfa, 2Department of Internal Medicine, Faculty of Medicine, Yuzuncu Yil University, Van, 3Department of Psychiatry, Faculty of Medicine, Kafkas University, Kars, 4Department of Clinical Biochemistry, Faculty of Medicine, Harran University, 5Department of Emergency Medicine, Faculty of Medicine, Harran University, Sanliurfa, 6Department of Psychiatry, Faculty of Medicine, Gaziantep University, Gaziantep, Turkey Background: As the relationship between psychological stress and platelet activation has been widely studied in recent years, activated platelets lead to certain biochemical changes, which occur in the brain in patients with mental disorders. However, data relating to the mean platelet volume (MPV in patients with panic disorder (PD are both limited and controversial. Herein, we aimed to evaluate, for the first time, the red cell distribution width (RDW levels combined with MPV levels in patients with PD.Patients and methods: Between January 2012 and June 2015, data of 30 treatment-naïve patients (16 females, 14 males; mean age: 37±10 years; range: 18–59 years who were diagnosed with PD and 25 age- and sex-matched healthy volunteers (10 females, 15 males; mean age: 36±13 years; range: 18–59 years (control group were retrospectively analyzed. The white blood cell count (WBC, MPV, and RDW levels were measured in both groups.Results: The mean WBC, MPV, and RDW levels were 9,173.03±2,400.31/mm3, 8.19±1.13 fl, and 12.47±1.14%, respectively, in the PD group. These values were found to be 7,090.24±1,032.61, 6.85±0.67, and 11.63±0.85, respectively, in the healthy controls. The WBC, MPV, and RDW levels were significantly higher in the patients with PD compared to the healthy controls (P=0.001, P=0.001, and P=0

  15. Comparison of imaging-based gross tumor volume and pathological volume determined by whole-mount serial sections in primary cervical cancer

    Directory of Open Access Journals (Sweden)

    Zhang Y

    2013-07-01

    Full Text Available Ying Zhang,1,* Jing Hu,1,* Jianping Li,1 Ning Wang,1 Weiwei Li,1 Yongchun Zhou,1 Junyue Liu,1 Lichun Wei,1 Mei Shi,1 Shengjun Wang,2 Jing Wang,2 Xia Li,3 Wanling Ma4 1Department of Radiation Oncology, 2Department of Nuclear Medicine, 3Department of Pathology, 4Department of Radiology, Xijing Hospital, Xi'an, People's Republic of China*These authors contributed equally to this workObjective: To investigate the accuracy of imaging-based gross tumor volume (GTV compared with pathological volume in cervical cancer.Methods: Ten patients with International Federation of Gynecology and Obstetrics stage I–II cervical cancer were eligible for investigation and underwent surgery in this study. Magnetic resonance imaging (MRI and fluorine-18 fluorodeoxyglucose positron emission tomography (18F-FDG PET/computed tomography (CT scans were taken the day before surgery. The GTVs under MRI and 18F-FDG PET/CT (GTV-MRI, GTV-PET, GTV-CT were calculated automatically by Eclipse treatment-planning systems. Specimens of excised uterine cervix and cervical cancer were consecutively sliced and divided into whole-mount serial sections. The tumor border of hematoxylin and eosin-stained sections was outlined under a microscope by an experienced pathologist. GTV through pathological image (GTV-path was calculated with Adobe Photoshop.Results: The GTVs (average ± standard deviation delineated and calculated under CT, MRI, PET, and histopathological sections were 19.41 ± 11.96 cm3, 12.66 ± 10.53 cm3, 11.07 ± 9.44 cm3, and 10.79 ± 8.71 cm3, respectively. The volume of GTV-CT or GTV-MR was bigger than GTV-path, and the difference was statistically significant (P 0.05. Spearman correlation analysis showed that GTV-CT, GTV-MRI, and GTV-PET were significantly correlated with GTV-path (P < 0.01. There was no significant difference in the lesion coverage factor among the three modalities.Conclusion: The present study showed that GTV defined under 40% of maximum standardized

  16. Acquired cisplatin resistance in human ovarian A2780 cancer cells correlates with shift in taurine homeostasis and ability to volume regulate

    DEFF Research Database (Denmark)

    Sørensen, Belinda Halling; Thorsteinsdottir, Unnur Arna; Lambert, Ian Henry

    2014-01-01

    Cisplatin resistance is a major challenge in the treatment of cancer and develops through reduced drug accumulation and an increased ability to avoid drug-induced cell damage, cell shrinkage, and hence initiation of apoptosis. Uptake and release of the semiessential amino acid taurine contribute...... to cell volume homeostasis, and taurine has been reported to have antiapoptotic effects. Here we find that volume-sensitive taurine release in cisplatin-sensitive [wild-type (WT)] human ovarian cancer A2780 cells is reduced in the presence of the phospholipase A2 inhibitor bromenol lactone, the 5......-induced cell death in RES A2780 cells correlates with an increased accumulation of taurine, due to an increased taurine uptake and a concomitant impairment of the volume-sensitive taurine release pathway, as well an inability to reduce cell volume after osmotic cell swelling. Downregulation of volume...

  17. A numerical-statistical approach to determining the representative elementary volume (REV of cement paste for measuring diffusivity

    Directory of Open Access Journals (Sweden)

    Zhang, M. Z.

    2010-12-01

    Full Text Available Concrete diffusivity is a function of its microstructure on many scales, ranging from nanometres to millimetres. Multi-scale techniques are therefore needed to model this parameter. Representative elementary volume (REV, in conjunction with the homogenization principle, is one of the most common multi-scale approaches. This study aimed to establish a procedure for establishing the REV required to determine cement paste diffusivity based on a three-step, numerical-statistical approach. First, several series of 3D cement paste microstructures were generated with HYMOSTRUC3D, a cement hydration and microstructure model, for different volumes of cement paste and w/c ratios ranging from 0.30 to 0.60. Second, the finite element method was used to simulate the diffusion of tritiated water through these microstructures. Effective cement paste diffusivity values for different REVs were obtained by applying Fick’s law. Finally, statistical analysis was used to find the fluctuation in effective diffusivity with cement paste volume, from which the REV was then determined. The conclusion drawn was that the REV for measuring diffusivity in cement paste is 100x100x100 μm3.

    La difusividad del hormigón depende de su microestructura a numerosas escalas, desde nanómetros hasta milímetros, por lo que se precisa de técnicas multiescala para representar este parámetro. Junto con el principio de homogeneización, uno de los métodos multiescala más habituales es el volumen elemental representativo (VER. El objeto de este estudio era establecer un procedimiento que permitiera determinar el VER necesario para calcular la difusividad de la pasta de cemento, basándose en un método numéricoestadístico que consta de tres etapas. Primero, se crearon varias series de microestructuras de pasta de cemento en 3D con HYMOSTRUC3D, un programa que permite crear un modelo de la hidratación y microestructura del cemento. Luego se empleó el método de

  18. [Dynamic expression profile of HBsAg according to hepatic parenchyma cells' volume at different liver fibrosis stages in the immune clearance phase].

    Science.gov (United States)

    Wu, Zhe-bin; Cao, Hong; Liu, Ting; Wu, Ze-qian; Ke, Wei-min; Gao, Zhi-liang

    2012-10-01

    The aim of this study was to determine the dynamic expression profile of hepatitis B surface antigen (HBsAg) according to hepatic parenchyma cells' volume at different stages of liver fibrosis during the immune clearance phase. Eighty-nine patients with HBeAg-positive chronic hepatitis B (CHB) in the immune clearance stage were recruited for study. Each patient's serum HBsAg levels were detected by electrochemiluminescence. The serum HBsAg levels were apportioned according to hepatic parenchyma cells' volume at liver fibrosis stages 1, 2, 3, and 4 and compared by ANOVA. The unapportioned serum HBsAg levels (IU/mL) at liver fibrosis stages 1 (227.2+/-237.7), 2 (211.0+/-131.4), 3(300.1+/-144.6), and 4 (278.7+/-148.8) were not significantly different (all comparisons, P range: 0.061 to 0.759). However, when the serum HBsAg levels were apportioned by the same hepatic parenchyma cells' volume at liver fibrosis stages 1 (343.9+/-359.8), 2 (336.4+/-209.5), 3 (508.7+/-245.1), and 4 (525.2+/-274.8), the levels were significantly different (all comparisons, F = 3.045 and P = 0.033; stage 1 vs. 3, P = 0.041; stage 1 vs. 4, P = 0.046; stage 2 vs. 3, P = 0.028; stage 2 vs. 4, P = 0.034). During the immune clearance phase of chronic hepatitis B, increased HBsAg expression is associated with increased hepatic parenchyma cells' volume and progressive liver fibrosis stage.

  19. Cell differentiation versus cell death: extracellular glucose is a key determinant of cell fate following oxidative stress exposure

    OpenAIRE

    Poulsen, R C; Knowles, H.J.; Carr, A. J.; HULLEY, P. A.

    2014-01-01

    Cells, particularly mechano-sensitive musculoskeletal cells such as tenocytes, routinely encounter oxidative stress. Oxidative stress can not only stimulate tissue repair, but also cause damage leading to tissue degeneration. As diabetes is associated with increased oxidative damage as well as increased risk of tendon degeneration, the aim of this study was to determine if extracellular glucose levels alter the response of tendon cells to oxidative stress. Primary human tenocytes were culture...

  20. Analysis of variance in determinations of equivalence volume and of the ionic product of water in potentiometric titrations.

    Science.gov (United States)

    Braibanti, A; Bruschi, C; Fisicaro, E; Pasquali, M

    1986-06-01

    Homogeneous sets of data from strong acid-strong base potentiometric titrations in aqueous solution at various constant ionic strengths have been analysed by statistical criteria. The aim is to see whether the error distribution matches that for the equilibrium constants determined by competitive potentiometric methods using the glass electrode. The titration curve can be defined when the estimated equivalence volume VEM, with standard deviation (s.d.) sigma (VEM), the standard potential E(0), with s.d. sigma(E(0)), and the operational ionic product of water K(*)(w) (or E(*)(w) in mV), with s.d. sigma(K(*)(w)) [or sigma(E(*)(w))] are known. A special computer program, BEATRIX, has been written which optimizes the values of VEM, E(0) and K(*)(w) by linearization of the titration curve as a Gran plot. Analysis of variance applied to a set of 11 titrations in 1.0M sodium chloride medium at 298 K has demonstrated that the values of VEM belong to a normal population of points corresponding to individual potential/volume data-pairs (E(i); v(i)) of any titration, whereas the values of pK(*)(w) (or of E(*)(w)) belong to a normal population with members corresponding to individual titrations, which is also the case for the equilibrium constants. The intertitration variation is attributable to the electrochemical component of the system and appears as signal noise distributed over the titrations. The correction for junction-potentials, introduced in a further stage of the program by optimization in a Nernst equation, increases the noise, i.e., sigma(pK(*)(w)). This correction should therefore be avoided whenever it causes an increase of sigma(pK(*)(w)). The influence of the ionic medium has been examined by processing data from acid-base titrations in 0.1M potassium chloride and 0.5M potassium nitrate media. The titrations in potassium chloride medium showed the same behaviour as those in sodium chloride medium, but with an s.d. for pK(*)(w) that was smaller and close to the

  1. Role of volume-regulated and calcium-activated anion channels in cell volume homeostasis, cancer and drug resistance

    DEFF Research Database (Denmark)

    Hoffmann, Else Kay; Sørensen, Belinda Halling; Sauter, Daniel Rafael Peter

    2015-01-01

    to be an essential component of both VRAC and VSOAC. Reduced VRAC and VSOAC activities are seen in drug resistant cancer cells. ANO1 is a calcium-activated chloride channel expressed on the plasma membrane of e.g. secretory epithelia. ANO1 is amplified and highly expressed in a large number of carcinomas. The gene...... functions as well as their role in cancer and drug resistance....

  2. Evaluation of the Efficacy of Standardized Uptake Value (SUV-shape Scheme for Thyroid Volume Determination in Graves’ Disease: A Comparison with Ultrasonography

    Directory of Open Access Journals (Sweden)

    yangchun chen

    2017-01-01

    Full Text Available Objective(s: In this study, we aimed to evaluate the efficacy of thyroid volume measurement using 99mTc pertechnetate single-photon emission computed tomography (SPECT images, acquired by the standardized uptake value (SUV-shape scheme designed by our expert team.Methods: A total of 18 consecutive patients with Graves’ disease (GD were subjected to both ultrasonographic and 99mTc pertechnetate SPECT examinations of thyroid within a five-day interval. The volume of thyroid lobes and isthmus was measured by ultrasonography (US according to the ellipsoid volume equation. The total thyroid volume, determined as the sum of the volume of both lobes and isthmus, was recorded as TV-US (i.e., thyroid volume measured by US and set as the reference. The thyroid volume was defined according to our SUV-shape scheme and was recorded as TV-SS (i.e., thyroid volume determined by the SUV-shape scheme. The data were analyzed using the Bland-Altman plot, linear regression analysis, Spearman’s rank correlation, and paired t-test, if necessary.Results: The values of TV-SS (40.2±29.4 mL and TV-US (43.0±34.7 mL were not significantly different (t=0.813; P=0.43. The linear regression equation of the two values was determined as TV-US= 1.072 × TV-SS − 0.29(r=0.906; P

  3. Determination of Shale Volume and Distribution Patterns and Effective Porosity from Well Log Data Based On Cross-Plot Approach for A Shaly Carbonate Gas Reservoir

    Science.gov (United States)

    Moradi, Siyamak; Moeini, Mohammad; Kamal Ghassem al-Askari, Mohammad; Hamed Mahvelati, Elaheh

    2016-10-01

    Determination of shale volume distribution is one of the most important factors that has to be considered in formation evaluation, since existence of shale reduces effective porosity and permeability of the reservoir. In this paper, shale volume and distribution (dispersed, laminar and structural) and formation effective porosity are estimated from well log data and cross-plots. Results show that distribution of shale is mainly dispersed with few of laminar ones, and the quality of reservoir (effective porosity) decreases with depth resulting in low productivity of gas wells drilled in lower zones. Good agreement of estimated shale volumes and effective porosities from neutron-density cross-plot with the values determined from gamma ray log (CGR) and core analysis demonstrates the accuracy and applicability of these plots in determination of petrophysical parameters from conventional log data.

  4. Determination of blood leukocyte concentration with constant volume acquisition on a flow cytometer is comparable to individualized single platform testing with beads as internal reference standard

    DEFF Research Database (Denmark)

    Hansen, Susan; Dahl, Ronald; Hoffmann, Hans Jürgen

    2008-01-01

    Flow cytometers have a constant flow rate. This enables flow cytometers to measure leukocyte concentrations in a determined volume by acquiring data at a fixed rate over a fixed time and is called constant volume acquisition (CVA). The volume aspirated by a FACS Calibur flow cytometer in 4 min...... at a high rate has a median of 163 microl (IQR 156-170) with TruCount tubes. Leukocyte concentrations of 26 healthy volunteers were measured twice on up to four occasions with a Bürker-Türk chamber, by single platform technology (SPT) with TruCount tubes and on the same data set using CVA. Total leukocyte...... concentrations determined by CVA correlated better with measurements in a Bürker-Türk (BT) chamber than with SPT. Concentrations determined with CVA were 1.86% higher than with BT whereas SPT data were 5.35% higher than BT (pCVA (p

  5. Prevention of cytotoxic T cell escape using a heteroclitic subdominant viral T cell determinant.

    Directory of Open Access Journals (Sweden)

    Noah S Butler

    2008-10-01

    Full Text Available High affinity antigen-specific T cells play a critical role during protective immune responses. Epitope enhancement can elicit more potent T cell responses and can subsequently lead to a stronger memory pool; however, the molecular basis of such enhancement is unclear. We used the consensus peptide-binding motif for the Major Histocompatibility Complex molecule H-2K(b to design a heteroclitic version of the mouse hepatitis virus-specific subdominant S598 determinant. We demonstrate that a single amino acid substitution at a secondary anchor residue (Q to Y at position 3 increased the stability of the engineered determinant in complex with H-2K(b. The structural basis for this enhanced stability was associated with local alterations in the pMHC conformation as a result of the Q to Y substitution. Recombinant viruses encoding this engineered determinant primed CTL responses that also reacted to the wildtype epitope with significantly higher functional avidity, and protected against selection of virus mutated at a second CTL determinant and consequent disease progression in persistently infected mice. Collectively, our findings provide a basis for the enhanced immunogenicity of an engineered determinant that will serve as a template for guiding the development of heteroclitic T cell determinants with applications in prevention of CTL escape in chronic viral infections as well as in tumor immunity.

  6. Cell-centered nonlinear finite-volume methods for the heterogeneous anisotropic diffusion problem

    Science.gov (United States)

    Terekhov, Kirill M.; Mallison, Bradley T.; Tchelepi, Hamdi A.

    2017-02-01

    We present two new cell-centered nonlinear finite-volume methods for the heterogeneous, anisotropic diffusion problem. The schemes split the interfacial flux into harmonic and transversal components. Specifically, linear combinations of the transversal vector and the co-normal are used that lead to significant improvements in terms of the mesh-locking effects. The harmonic component of the flux is represented using a conventional monotone two-point flux approximation; the component along the parameterized direction is treated nonlinearly to satisfy either positivity of the solution as in [29], or the discrete maximum principle as in [9]. In order to make the method purely cell-centered, we derive a homogenization function that allows for seamless interpolation in the presence of heterogeneity following a strategy similar to [46]. The performance of the new schemes is compared with existing multi-point flux approximation methods [3,5]. The robustness of the scheme with respect to the mesh-locking problem is demonstrated using several challenging test cases.

  7. Mast cell blocking reduces brain edema and hematoma volume and improves outcome after experimental intracerebral hemorrhage.

    Science.gov (United States)

    Strbian, Daniel; Tatlisumak, Turgut; Ramadan, Usama Abo; Lindsberg, Perttu J

    2007-04-01

    Intracerebral hemorrhage (ICH) is associated with high mortality and disability, and there is no widely approved clinical therapy. Poor outcome after ICH results mostly from a mass effect owing to enlargement of the hematoma and brain swelling, leading to displacement and disruption of brain structures. Cerebral mast cells (MC) are resident inflammatory cells that are located perivascularly and contain potent vasoactive, proteolytic, and fibrinolytic substances. We previously found pharmacological MC stabilization and genetic MC deficiency to be associated with up to 50% reduction of postischemic brain swelling in rats. Here, we studied the role of MC and MC stabilization in ICH using in vivo magnetic resonance imaging and ex vivo digital imaging for calculating brain edema and hematoma volume. In a rat ICH model of autologous blood injection into the basal ganglia, four groups of Wistar rats received either saline or sodium cromoglycate (MC stabilizer, two groups) or compound 48/80 (MC degranulator). Evaluated 24 h later, MC stabilization had resulted in highly significantly better neurologic scores (Pbrain swelling (Pbrain swelling (P<0.05), and smaller hematoma growth (P<0.05) than WT. The role of MC deserves a close evaluation as a potential target in the development of novel forms of ICH therapy.

  8. Natural biased coin encoded in the genome determines cell strategy.

    Science.gov (United States)

    Dorri, Faezeh; Mahini, Hamid; Sharifi-Zarchi, Ali; Totonchi, Mehdi; Tusserkani, Ruzbeh; Pezeshk, Hamid; Sadeghi, Mehdi

    2014-01-01

    Decision making at a cellular level determines different fates for isogenic cells. However, it is not yet clear how rational decisions are encoded in the genome, how they are transmitted to their offspring, and whether they evolve and become optimized throughout generations. In this paper, we use a game theoretic approach to explain how rational decisions are made in the presence of cooperators and competitors. Our results suggest the existence of an internal switch that operates as a biased coin. The biased coin is, in fact, a biochemical bistable network of interacting genes that can flip to one of its stable states in response to different environmental stimuli. We present a framework to describe how the positions of attractors in such a gene regulatory network correspond to the behavior of a rational player in a competing environment. We evaluate our model by considering lysis/lysogeny decision making of bacteriophage lambda in E. coli.

  9. Natural biased coin encoded in the genome determines cell strategy.

    Directory of Open Access Journals (Sweden)

    Faezeh Dorri

    Full Text Available Decision making at a cellular level determines different fates for isogenic cells. However, it is not yet clear how rational decisions are encoded in the genome, how they are transmitted to their offspring, and whether they evolve and become optimized throughout generations. In this paper, we use a game theoretic approach to explain how rational decisions are made in the presence of cooperators and competitors. Our results suggest the existence of an internal switch that operates as a biased coin. The biased coin is, in fact, a biochemical bistable network of interacting genes that can flip to one of its stable states in response to different environmental stimuli. We present a framework to describe how the positions of attractors in such a gene regulatory network correspond to the behavior of a rational player in a competing environment. We evaluate our model by considering lysis/lysogeny decision making of bacteriophage lambda in E. coli.

  10. A robust method for pulse peak determination in a digital volume pulse waveform with a wandering baseline.

    Science.gov (United States)

    Jang, Dae-Geun; Farooq, Umar; Park, Seung-Hun; Hahn, Minsoo

    2014-10-01

    This paper presents a robust method for pulse peak determination in a digital volume pulse (DVP) waveform with a wandering baseline. A proposed new method uses a modified morphological filter (MMF) to eliminate a wandering baseline signal of the DVP signal with minimum distortion and a slope sum function (SSF) with an adaptive thresholding scheme to detect pulse peaks from the baseline-removed DVP signal. Further in order to cope with over-detected and missed pulse peaks, knowledge based rules are applied as a postprocessor. The algorithm automatically adjusts detection parameters periodically to adapt to varying beat morphologies and fluctuations. Compared with conventional methods (highpass filtering, linear interpolation, cubic spline interpolation, and wavelet adaptive filtering), our method performs better in terms of the signal-to-error ratio, the computational burden (0.125 seconds for one minute of DVP signal analysis with the Intel Core 2 Quad processor @ 2.40 GHz PC), the true detection rate (97.32% with an acceptance level of 4 ms ) as well as the normalized error rate (0.18%). In addition, the proposed method can detect true positions of pulse peaks more accurately and becomes very useful for pulse transit time (PTT) and pulse rate variability (PRV) analyses.

  11. MGA: A gamma-ray spectrum analysis code for determining plutonium isotopic abundances. Volume 3, FORTRAN listing of the GA code

    Energy Technology Data Exchange (ETDEWEB)

    Gunnink, R

    1991-09-01

    Nondestructive measurements of x-ray and gamma-ray emissions can be used to determine the abundances of various actinides in a sample. Volume 1 of this report describes the methods and algorithms we have developed to determine the relative isotopic abundances of actinides in a sample, by analyzing gamma-ray spectra obtained using germanium detector systems. Volume 2 is a guide to using the MGA (Multiple Group Analysis) computer program we have written to perform plutonium isotopic analyses. This report contains a listing of the FORTRAN instructions of the code.

  12. Factors influencing the determination of cell traction forces.

    Science.gov (United States)

    Zündel, Manuel; Ehret, Alexander E; Mazza, Edoardo

    2017-01-01

    Methods summarized by the term Traction Force Microscopy are widely used to quantify cellular forces in mechanobiological studies. These methods are inverse, in the sense that forces must be determined such that they comply with a measured displacement field. This study investigates how several experimental and analytical factors, originating in the realization of the experiments and the procedures for the analysis, affect the determined traction forces. The present results demonstrate that even for very high resolution measurements free of noise, traction forces can be significantly underestimated, while traction peaks are typically overestimated by 50% or more, even in the noise free case. Compared to this errors, which are inherent to the nature of the mechanical problem and its discretization, the effect of ignoring the out-of-plane displacement component, the interpolation scheme used between the discrete measurement points and the disregard of the geometrical non-linearities when using a nearly linear substrate material are less consequential. Nevertheless, a nonlinear elastic substrate, with strain-stiffening response and some degree of compressibility, can substantially improve the robustness of the reconstruction of traction forces over a wide range of magnitudes. This poses the need for a correct mechanical representation of these non-linearities during the traction reconstruction and a correct mechanical characterization of the substrate itself, especially for the large strain shear domain which is shown to plays a major role in the deformations induced by cells.

  13. Determination of saturation pressure and enthalpy of vaporization of semi-volatile aerosols: the integrated volume mentod

    Science.gov (United States)

    This study presents the integrated volume method for estimating saturation pressure and enthalpy of vaporization of a whole aerosol distribution. We measure the change of total volume of an aerosol distribution between a reference state and several heated states, with the heating...

  14. Reliability of In Vivo Determination of Forearm Muscle Volume Using 3.0 T Magnetic Resonance Imaging

    NARCIS (Netherlands)

    M.J.C. Smeulders; S. van den Berg; J. Oudeman; A.J. Nederveen; M. Kreulen; M. Maas

    2010-01-01

    Purpose: To apply magnetic resonance imaging (MRI) as a tool for quantifying muscle volume of forearm muscles feasibility and reliability of volume estimation of the flexor carpi ulnaris (FCU) and the extensor carpi ulnaris (ECU). Materials and Methods: Forearms of 10 subjects were scanned twice. Mu

  15. Multiplexing spheroid volume, resazurin and acid phosphatase viability assays for high-throughput screening of tumour spheroids and stem cell neurospheres.

    Science.gov (United States)

    Ivanov, Delyan P; Parker, Terry L; Walker, David A; Alexander, Cameron; Ashford, Marianne B; Gellert, Paul R; Garnett, Martin C

    2014-01-01

    Three-dimensional cell culture has many advantages over monolayer cultures, and spheroids have been hailed as the best current representation of small avascular tumours in vitro. However their adoption in regular screening programs has been hindered by uneven culture growth, poor reproducibility and lack of high-throughput analysis methods for 3D. The objective of this study was to develop a method for a quick and reliable anticancer drug screen in 3D for tumour and human foetal brain tissue in order to investigate drug effectiveness and selective cytotoxic effects. Commercially available ultra-low attachment 96-well round-bottom plates were employed to culture spheroids in a rapid, reproducible manner amenable to automation. A set of three mechanistically different methods for spheroid health assessment (Spheroid volume, metabolic activity and acid phosphatase enzyme activity) were validated against cell numbers in healthy and drug-treated spheroids. An automated open-source ImageJ macro was developed to enable high-throughput volume measurements. Although spheroid volume determination was superior to the other assays, multiplexing it with resazurin reduction and phosphatase activity produced a richer picture of spheroid condition. The ability to distinguish between effects on malignant and the proliferating component of normal brain was tested using etoposide on UW228-3 medulloblastoma cell line and human neural stem cells. At levels below 10 µM etoposide exhibited higher toxicity towards proliferating stem cells, whereas at concentrations above 10 µM the tumour spheroids were affected to a greater extent. The high-throughput assay procedures use ready-made plates, open-source software and are compatible with standard plate readers, therefore offering high predictive power with substantial savings in time and money.

  16. Multiplexing spheroid volume, resazurin and acid phosphatase viability assays for high-throughput screening of tumour spheroids and stem cell neurospheres.

    Directory of Open Access Journals (Sweden)

    Delyan P Ivanov

    Full Text Available Three-dimensional cell culture has many advantages over monolayer cultures, and spheroids have been hailed as the best current representation of small avascular tumours in vitro. However their adoption in regular screening programs has been hindered by uneven culture growth, poor reproducibility and lack of high-throughput analysis methods for 3D. The objective of this study was to develop a method for a quick and reliable anticancer drug screen in 3D for tumour and human foetal brain tissue in order to investigate drug effectiveness and selective cytotoxic effects. Commercially available ultra-low attachment 96-well round-bottom plates were employed to culture spheroids in a rapid, reproducible manner amenable to automation. A set of three mechanistically different methods for spheroid health assessment (Spheroid volume, metabolic activity and acid phosphatase enzyme activity were validated against cell numbers in healthy and drug-treated spheroids. An automated open-source ImageJ macro was developed to enable high-throughput volume measurements. Although spheroid volume determination was superior to the other assays, multiplexing it with resazurin reduction and phosphatase activity produced a richer picture of spheroid condition. The ability to distinguish between effects on malignant and the proliferating component of normal brain was tested using etoposide on UW228-3 medulloblastoma cell line and human neural stem cells. At levels below 10 µM etoposide exhibited higher toxicity towards proliferating stem cells, whereas at concentrations above 10 µM the tumour spheroids were affected to a greater extent. The high-throughput assay procedures use ready-made plates, open-source software and are compatible with standard plate readers, therefore offering high predictive power with substantial savings in time and money.

  17. Local positive feedback regulation determines cell shape in root hair cells.

    Science.gov (United States)

    Takeda, Seiji; Gapper, Catherine; Kaya, Hidetaka; Bell, Elizabeth; Kuchitsu, Kazuyuki; Dolan, Liam

    2008-02-29

    The specification and maintenance of growth sites are tightly regulated during cell morphogenesis in all organisms. ROOT HAIR DEFECTIVE 2 reduced nicotinamide adenine dinucleotide phosphate (RHD2 NADPH) oxidase-derived reactive oxygen species (ROS) stimulate a Ca2+ influx into the cytoplasm that is required for root hair growth in Arabidopsis thaliana. We found that Ca2+, in turn, activated the RHD2 NADPH oxidase to produce ROS at the growing point in the root hair. Together, these components could establish a means of positive feedback regulation that maintains an active growth site in expanding root hair cells. Because the location and stability of growth sites predict the ultimate form of a plant cell, our findings demonstrate how a positive feedback mechanism involving RHD2, ROS, and Ca2+ can determine cell shape.

  18. Disorders of Erythrocyte Volume Homeostasis

    OpenAIRE

    Glogowska, Edyta; Gallagher, Patrick G.

    2015-01-01

    Inherited disorders of erythrocyte volume homeostasis are a heterogeneous group of rare disorders with phenotypes ranging from dehydrated to overhydrated erythrocytes. Clinical, laboratory, physiologic, and genetic heterogeneity characterize this group of disorders. A series of recent reports have provided novel insights into our understanding of the genetic bases underlying some of these disorders of red cell volume regulation. This report reviews this progress in understanding determinants ...

  19. Differentiation state determines neural effects on microvascular endothelial cells

    Energy Technology Data Exchange (ETDEWEB)

    Muffley, Lara A., E-mail: muffley@u.washington.edu [University of Washington, Campus Box 359796, 300 9th Avenue, Seattle, WA 98104 (United States); Pan, Shin-Chen, E-mail: pansc@mail.ncku.edu.tw [University of Washington, Campus Box 359796, 300 9th Avenue, Seattle, WA 98104 (United States); Smith, Andria N., E-mail: gnaunderwater@gmail.com [University of Washington, Campus Box 359796, 300 9th Avenue, Seattle, WA 98104 (United States); Ga, Maricar, E-mail: marga16@uw.edu [University of Washington, Campus Box 359796, 300 9th Avenue, Seattle, WA 98104 (United States); Hocking, Anne M., E-mail: ahocking@u.washington.edu [University of Washington, Campus Box 359796, 300 9th Avenue, Seattle, WA 98104 (United States); Gibran, Nicole S., E-mail: nicoleg@u.washington.edu [University of Washington, Campus Box 359796, 300 9th Avenue, Seattle, WA 98104 (United States)

    2012-10-01

    Growing evidence indicates that nerves and capillaries interact paracrinely in uninjured skin and cutaneous wounds. Although mature neurons are the predominant neural cell in the skin, neural progenitor cells have also been detected in uninjured adult skin. The aim of this study was to characterize differential paracrine effects of neural progenitor cells and mature sensory neurons on dermal microvascular endothelial cells. Our results suggest that neural progenitor cells and mature sensory neurons have unique secretory profiles and distinct effects on dermal microvascular endothelial cell proliferation, migration, and nitric oxide production. Neural progenitor cells and dorsal root ganglion neurons secrete different proteins related to angiogenesis. Specific to neural progenitor cells were dipeptidyl peptidase-4, IGFBP-2, pentraxin-3, serpin f1, TIMP-1, TIMP-4 and VEGF. In contrast, endostatin, FGF-1, MCP-1 and thrombospondin-2 were specific to dorsal root ganglion neurons. Microvascular endothelial cell proliferation was inhibited by dorsal root ganglion neurons but unaffected by neural progenitor cells. In contrast, microvascular endothelial cell migration in a scratch wound assay was inhibited by neural progenitor cells and unaffected by dorsal root ganglion neurons. In addition, nitric oxide production by microvascular endothelial cells was increased by dorsal root ganglion neurons but unaffected by neural progenitor cells. -- Highlights: Black-Right-Pointing-Pointer Dorsal root ganglion neurons, not neural progenitor cells, regulate microvascular endothelial cell proliferation. Black-Right-Pointing-Pointer Neural progenitor cells, not dorsal root ganglion neurons, regulate microvascular endothelial cell migration. Black-Right-Pointing-Pointer Neural progenitor cells and dorsal root ganglion neurons do not effect microvascular endothelial tube formation. Black-Right-Pointing-Pointer Dorsal root ganglion neurons, not neural progenitor cells, regulate

  20. Relationship between blood group, packed cell volume, knowledge and practices of adults to malaria prevalence in Lagos Island, Nigeria

    Institute of Scientific and Technical Information of China (English)

    Bamidele Akinsanya; Adedotun Adesegun Adenusi; Oluwatobi Dolapo

    2016-01-01

    Objective:To determine malaria prevalence, knowledge, attitude and practices at residents of Lagos Island Local Government. Methods: Two hundred blood samples were collected from adults attending the General Hospital, Marina while 100 questionnaires were administered to individuals still in the same hospital. Majority of the respondents were between the ages of 18 and 30 years (44%), while the age group 60 and above had the lowest population (9%). Results:More than half of the respondents (68%) were employed and engaged in different occupations such as entrepreneur (32%), students (31%), trader (24%), and civil servant (10%). Stratification of the respondents by income revealed that 71% earned above the minimum wage, while 29% earned below. The prevalence of malaria by microscopy method was 10.5%. Malaria parasite had significant effect on the packed cell volume of infected individuals (P < 0.05). Preventive measures employed by the respondents were majorly insecticide (72.2%), while some others (5.6%) used mosquito net. Majority of the respondents (98%) believed that malaria was caused by mosquito. Conclusions: Therefore, it is appropriate to put in place preventive measures against malaria to avoid high prevalence of the number one killer in tropical Africa.

  1. Pretreatment metabolic tumour volume is predictive of disease-free survival and overall survival in patients with oesophageal squamous cell carcinoma

    Energy Technology Data Exchange (ETDEWEB)

    Lemarignier, Charles; Gouel, Pierrick [Centre Henri Becquerel and Rouen University Hospital, Department of Nuclear Medicine, Rouen (France); Di Fiore, Frederic [Rouen University Hospital, Department of Gastroenterology, Rouen (France); Centre Henri Becquerel, Department of Medical oncology, Rouen (France); Marre, Charline; Michel, Pierre [Rouen University Hospital, Department of Gastroenterology, Rouen (France); Hapdey, Sebastien; Modzelewski, Romain; Vera, Pierre [Centre Henri Becquerel and Rouen University Hospital, Department of Nuclear Medicine, Rouen (France); University of Rouen, QuantIF (Litis EA 4108 - FR CNRS 3638), Rouen (France); Dubray, Bernard [University of Rouen, QuantIF (Litis EA 4108 - FR CNRS 3638), Rouen (France); Centre Henri Becquerel and Rouen University Hospital, Department of Radiotherapy and Medical Physics, Rouen (France)

    2014-11-15

    It has been suggested that FDG PET has predictive value for the prognosis of treated oesophageal carcinoma. However, the studies reported in the literature have shown discordant results. The aim of this study was to determine whether pretherapy quantitative metabolic parameters correlate with patient outcomes. Included in the study were 67 patients with a histological diagnosis of oesophageal squamous cell carcinoma. Each patient underwent {sup 18}F-FDG PET (4.5 MBq/kg) before chemoradiotherapy. Quantitative analysis was performed using the following parameters: age, weight loss, location, N stage, OMS performance status, MTV{sub p} and MTV{sub p'} (metabolic tumour volume determined by two different physicians), MTV{sub 40%} (volume for a threshold of 40 % of SUVmax), MTV{sub a} (volume automatically determined with a contrast-based adaptive threshold method), SUVmax, SUVmean and TLG (total lesion glycolysis). MTV{sub p} and MTV{sub 40%} were highly correlated (Pearson's index 0.92). SUVmean{sub p} and SUVmean{sub 40%} were also correlated (Pearson's index 0.86), as were TLG{sub p} and TLG{sub 40%} (Pearson's index 0.98). Similarly, the parameters obtained with the adaptive threshold method (MTV{sub a}, SUVmean{sub a} and TLG{sub a}) were correlated with those obtained manually (MTV{sub p}, SUVmean{sub p} and TLG{sub p}). The manual metabolic tumour volume determination (MTV{sub p} and MTV{sub p'}) was reproducible. Multivariate analysis for disease-free survival (DFS) showed that a larger MTV{sub p} was associated with a shorter DFS (p = 0.004) and that a higher SUVmax was associated with a longer DFS (p = 0.02). Multivariate analysis for overall survival (OS) showed that a larger MTV{sub p} was associated with a shorter OS (p = 0.01) and that a tumour in the distal oesophagus was associated with a longer OS (p = 0.005). The associations among the other parameters were not statistically significant. Metabolic tumour volume is a major

  2. A GIS-based method to determine the volume of lahars: Popocatépetl volcano, Mexico

    Science.gov (United States)

    Muñoz-Salinas, E.; Renschler, C. S.; Palacios, D.

    2009-10-01

    Lahars are flows composed of water and volcanic sediment which are often dangerous for people living near volcanoes. Therefore, a reliable estimation of lahar volume is needed to effectively assess the risk. This paper proposes a new method to calculate the volume of lahar sediments found in channels of volcanic landscapes. The method requires surveys of several cross-sections along a gorge, a Digital Elevation Model of the study area and measurements of the thickness of the lahar deposits. With these data and a Geographical Information System (GIS), the volume is calculated for the erosive section where deposit volume is divided into oblique parallelepipeds and the sedimentary section where deposit volume is divided into polyhedrons. This new method was applied to the 1997 and 2001 lahars that occurred in the channel of a gorge at Popocatépetl volcano, Mexico. The estimated volumes are 1.85 × 10 5 and 1.6 × 10 5 m 3, respectively, which is about 40% less than those obtained by the traditional method that multiplies lahar flow-path length, sediment width and sediment depth. This observation suggests that the traditional method tends to overestimate volumes.

  3. Genetic determinants and stroke in children with sickle cell disease.

    Science.gov (United States)

    Rodrigues, Daniela O W; Ribeiro, Luiz C; Sudário, Lysla C; Teixeira, Maria T B; Martins, Marina L; Pittella, Anuska M O L; Junior, Irtis de O Fernandes

    To verify genetic determinants associated with stroke in children with sickle cell disease (SCD). Prospective cohort with 110 children submitted to neonatal screening by the Neonatal Screening Program, between 1998 and 2007, with SCD diagnosis, followed at a regional reference public service for hemoglobinopathies. The analyzed variables were type of hemoglobinopathy, gender, coexistence with alpha thalassemia (α-thal), haplotypes of the beta globin chain cluster, and stroke. The final analysis was conducted with 66 children with sickle cell anemia (SCA), using the chi-squared test in the program SPSS(®) version 14.0. Among children with SCD, 60% had SCA. The prevalence of coexistence with α-thal was 30.3% and the Bantu haplotype (CAR) was identified in 89.2%. The incidence of stroke was significantly higher in those with SCA (27.3% vs. 2.3%; p=0.001) and males (24.1% vs. 9.6%; p=0.044). The presence of α-thal (p=0.196), the CAR haplotype (p=0.543), and socioeconomic factors were not statistically significant in association with the occurrence of stroke. There is a high incidence of stroke in male children and in children with SCA. Coexistence with α-thal and haplotypes of the beta globin chain cluster did not show any significant association with stroke. The heterogeneity between previously evaluated populations, the non-reproducibility between studies, and the need to identify factors associated with stroke in patients with SCA indicate the necessity of conducting further research to demonstrate the relevance of genetic factors in stroke related to SCD. Copyright © 2016 Sociedade Brasileira de Pediatria. Published by Elsevier Editora Ltda. All rights reserved.

  4. Genetic determinants and stroke in children with sickle cell disease,

    Directory of Open Access Journals (Sweden)

    Daniela O.W. Rodrigues

    Full Text Available Abstract Objective: To verify genetic determinants associated with stroke in children with sickle cell disease (SCD. Methods: Prospective cohort with 110 children submitted to neonatal screening by the Neonatal Screening Program, between 1998 and 2007, with SCD diagnosis, followed at a regional reference public service for hemoglobinopathies. The analyzed variables were type of hemoglobinopathy, gender, coexistence with alpha thalassemia (α-thal, haplotypes of the beta globin chain cluster, and stroke. The final analysis was conducted with 66 children with sickle cell anemia (SCA, using the chi-squared test in the program SPSS® version 14.0. Results: Among children with SCD, 60% had SCA. The prevalence of coexistence with α-thal was 30.3% and the Bantu haplotype (CAR was identified in 89.2%. The incidence of stroke was significantly higher in those with SCA (27.3% vs. 2.3%; p = 0.001 and males (24.1% vs. 9.6%; p = 0.044. The presence of α-thal (p = 0.196, the CAR haplotype (p = 0.543, and socioeconomic factors were not statistically significant in association with the occurrence of stroke. Conclusion: There is a high incidence of stroke in male children and in children with SCA. Coexistence with α-thal and haplotypes of the beta globin chain cluster did not show any significant association with stroke. The heterogeneity between previously evaluated populations, the non-reproducibility between studies, and the need to identify factors associated with stroke in patients with SCA indicate the necessity of conducting further research to demonstrate the relevance of genetic factors in stroke related to SCD.

  5. Systematic determination of thyroid volume by ultrasound examination from infancy to adolescence in Japan: the Fukushima Health Management Survey.

    Science.gov (United States)

    Suzuki, Satoru; Midorikawa, Sanae; Fukushima, Toshihiko; Shimura, Hiroki; Ohira, Tetsuya; Ohtsuru, Akira; Abe, Masafumi; Shibata, Yoshisada; Yamashita, Shunichi; Suzuki, Shinichi

    2015-01-01

    Although several reports have defined normal thyroid volume depending on either age or body surface, there are no sequential reference values on childhood thyroid volume evaluated by using ultrasonography and epidemiological analysis in Japan. The aim of the present study was to establish updated reference values for thyroid volume by ultrasound examination and epidemiological analysis in 0-19 year-old Japanese children. It is based on a cross-sectional study conducted from October 9, 2011 to March 31, 2012. The subjects were 38,063 children who were examined by ultrasonography as the initial preliminary survey of the Fukushima Health Management Survey in October 9, 2011 to March 31, 2012. The width, thickness, and height of each lobe were measured and the volume of each lobe was calculated by the mean of the elliptical shape volume formula. The values of thyroid volume at the 2.5 and 97.5 percentiles of age and body surface area for each gender group were obtained from 0-19 year-old children. Positive correlation was observed between thyroid volume and either age or body surface. The right lobe was significantly larger than the left lobe. The thyroid volume in females was larger than that in males after adjusting body surface area. The reference values of childhood thyroid for each age or body surface area were obtained by this extensive survey using ultrasound. These reference values may be used to define the normal size of thyroid gland by echosonography in Japanese children, although thyroid volume may be affected by dimorphic factors such as sex hormones.

  6. Dynamic Regulation of Cell Volume and Extracellular ATP of Human Erythrocytes

    Science.gov (United States)

    Leal Denis, M. Florencia; Alvarez, H. Ariel; Lauri, Natalia; Alvarez, Cora L.; Chara, Osvaldo; Schwarzbaum, Pablo J.

    2016-01-01

    Introduction The peptide mastoparan 7 (MST7) triggered in human erythrocytes (rbcs) the release of ATP and swelling. Since swelling is a well-known inducer of ATP release, and extracellular (ATPe), interacting with P (purinergic) receptors, can affect cell volume (Vr), we explored the dynamic regulation between Vr and ATPe. Methods and Treatments We made a quantitative assessment of MST7-dependent kinetics of Vr and of [ATPe], both in the absence and presence of blockers of ATP efflux, swelling and P receptors. Results In rbcs 10 μM MST7 promoted acute, strongly correlated changes in [ATPe] and Vr. Whereas MST7 induced increases of 10% in Vr and 190 nM in [ATPe], blocking swelling in a hyperosmotic medium + MST7 reduced [ATPe] by 40%. Pre-incubation of rbcs with 10 μM of either carbenoxolone or probenecid, two inhibitors of the ATP conduit pannexin 1, reduced [ATPe] by 40–50% and swelling by 40–60%, while in the presence of 80 U/mL apyrase, an ATPe scavenger, cell swelling was prevented. While exposure to 10 μM NF110, a blocker of ATP-P2X receptors mediating sodium influx, reduced [ATPe] by 48%, and swelling by 80%, incubation of cells in sodium free medium reduced swelling by 92%. Analysis and Discussion Results were analyzed by means of a mathematical model where ATPe kinetics and Vr kinetics were mutually regulated. Model dependent fit to experimental data showed that, upon MST7 exposure, ATP efflux required a fast 1960-fold increase of ATP permeability, mediated by two kinetically different conduits, both of which were activated by swelling and inactivated by time. Both experimental and theoretical results suggest that, following MST7 exposure, ATP is released via two conduits, one of which is mediated by pannexin 1. The accumulated ATPe activates P2X receptors, followed by sodium influx, resulting in cell swelling, which in turn further activates ATP release. Thus swelling and P2X receptors constitute essential components of a positive feedback loop

  7. Dynamic Regulation of Cell Volume and Extracellular ATP of Human Erythrocytes.

    Directory of Open Access Journals (Sweden)

    M Florencia Leal Denis

    Full Text Available The peptide mastoparan 7 (MST7 triggered in human erythrocytes (rbcs the release of ATP and swelling. Since swelling is a well-known inducer of ATP release, and extracellular (ATPe, interacting with P (purinergic receptors, can affect cell volume (Vr, we explored the dynamic regulation between Vr and ATPe.We made a quantitative assessment of MST7-dependent kinetics of Vr and of [ATPe], both in the absence and presence of blockers of ATP efflux, swelling and P receptors.In rbcs 10 μM MST7 promoted acute, strongly correlated changes in [ATPe] and Vr. Whereas MST7 induced increases of 10% in Vr and 190 nM in [ATPe], blocking swelling in a hyperosmotic medium + MST7 reduced [ATPe] by 40%. Pre-incubation of rbcs with 10 μM of either carbenoxolone or probenecid, two inhibitors of the ATP conduit pannexin 1, reduced [ATPe] by 40-50% and swelling by 40-60%, while in the presence of 80 U/mL apyrase, an ATPe scavenger, cell swelling was prevented. While exposure to 10 μM NF110, a blocker of ATP-P2X receptors mediating sodium influx, reduced [ATPe] by 48%, and swelling by 80%, incubation of cells in sodium free medium reduced swelling by 92%.Results were analyzed by means of a mathematical model where ATPe kinetics and Vr kinetics were mutually regulated. Model dependent fit to experimental data showed that, upon MST7 exposure, ATP efflux required a fast 1960-fold increase of ATP permeability, mediated by two kinetically different conduits, both of which were activated by swelling and inactivated by time. Both experimental and theoretical results suggest that, following MST7 exposure, ATP is released via two conduits, one of which is mediated by pannexin 1. The accumulated ATPe activates P2X receptors, followed by sodium influx, resulting in cell swelling, which in turn further activates ATP release. Thus swelling and P2X receptors constitute essential components of a positive feedback loop underlying ATP-induced ATP release of rbcs.

  8. Sphaeropsidin A shows promising activity against drug-resistant cancer cells by targeting regulatory volume increase

    Science.gov (United States)

    Mathieu, Véronique; Chantôme, Aurélie; Lefranc, Florence; Cimmino, Alessio; Miklos, Walter; Paulitschke, Verena; Mohr, Thomas; Maddau, Lucia; Kornienko, Alexander; Berger, Walter; Vandier, Christophe; Evidente, Antonio; Delpire, Eric; Kiss, Robert

    2016-01-01

    Despite the recent advances in the treatment of tumors with intrinsic chemotherapy resistance, such as melanoma and renal cancers, their prognosis remains poor and new chemical agents with promising activity against these cancers are urgently needed. Sphaeropsidin A, a fungal metabolite whose anticancer potential had previously received little attention, was isolated from Diplodia cupressi and found to display specific anticancer activity in vitro against melanoma and kidney cancer subpanels in the National Cancer Institute (NCI) 60-cell line screen. The NCI data revealed a mean LC50 of ca. 10 μM and a cellular sensitivity profile that did not match that of any other agent in the 765,000 compound database. Subsequent mechanistic studies in melanoma and other multidrug-resistant in vitro cancer models showed that sphaeropsidin A can overcome apoptosis as well as multidrug resistance by inducing a marked and rapid cellular shrinkage related to the loss of intracellular Cl− and the decreased HCO3− concentration in the culture supernatant. These changes in ion homeostasis and the absence of effects on the plasma membrane potential were attributed to the sphaeropsidin A-induced impairment of regulatory volume increase (RVI). Preliminary results also indicate that depending on the type of cancer, the sphaeropsidin A effects on RVI could be related to Na–K–2Cl electroneutral cotransporter or Cl−/HCO3− anion exchanger(s) targeting. This study underscores the modulation of ion-transporter activity as a promising therapeutic strategy to combat drug-resistant cancers and identifies the fungal metabolite, sphaeropsidin A, as a lead to develop anticancer agents targeting RVI in cancer cells. PMID:25868554

  9. Red cell distribution width and mean platelet volume in patients with irritable bowel syndrome

    Science.gov (United States)

    Alcelik, Aytekin; Tekce, Buket Kin; Tekelioglu, Vildan; Sit, Mustafa; Savli, Haluk

    2014-01-01

    Introduction Possible pathophysiological mechanisms of irritable bowel syndrome (IBS) are interactions between microbial flora of the gut and the mucosal/systemic immune system, post-infectious status and inflammation. Mean platelet volume (MPV) and red cell distribution width (RDW) have been reported as inflammatory markers in patients with inflammatory bowel disease, but they have not been studied in functional gastrointestinal disorders. Aim To investigate whether there was an association between haemogram parameters (RDW and MPV) and IBS. Material and methods Forty patients with IBS and 44 healthy controls were included to this retrospective study. Patients diagnosed with IBS according to Rome III criteria were included as the IBS group. They were all screened for psychiatric or organic bowel diseases for the sake of precise diagnosis. Results Both RDW (p < 0.001) and MPV (p = 0.046) were increased in patients with IBS compared to controls. This increase in RDW and MPV was independent of the type of IBS. Conclusions The RDW and MPV should be laboratory indicators of IBS. More prospective studies with larger cohorts are needed to confirm our results. PMID:25097713

  10. Water equilibria and management using a two-volume model of a polymer electrolyte fuel cell

    Science.gov (United States)

    Karnik, Amey Y.; Stefanopoulou, Anna G.; Sun, Jing

    In this paper, we introduce a modified interpretation of the water activity presented in Springer et al. [T.E. Springer, T.A. Zawodzinski, S. Gottesfeld, Polymer electrolyte fuel cell model, J. Electrochem. Soc. 138 (8) (1991) 2334-2342]. The modification directly affects the membrane water transport between the anode and the cathode (two electrodes) of the polymer electrolyte membrane (PEM) fuel cell in the presence of liquid water inside the stack. The modification permits calibration of a zero-dimensional isothermal model to predict the flooding and drying conditions in the two electrodes observed at various current levels [D. Spernjak, S. Advani, A.K. Prasad, Experimental investigation of liquid water formation and transport in a transparent single-serpentine PEM fuel cell, in: Proceedings of the Fourth International Conference on Fuel Cell Science, Engineering and Technology (FUELCELL2006-97271), June 2006]. Using this model the equilibria of the lumped water mass in the two electrodes are analyzed at various flow conditions of the stack to determine stable and unstable (liquid water growth) operating conditions. Two case studies of water management through modification of cathode inlet humidification and anode water removal are then evaluated using this model. The desired anode water removal and the desired cathode inlet humidification are specified based upon (i) the water balance requirements, (ii) the desired conditions in the electrodes, and (iii) the maximum membrane transport at those conditions.

  11. Bone volume fraction and fabric anisotropy are better determinants of trabecular bone stiffness than other morphological variables.

    Science.gov (United States)

    Maquer, Ghislain; Musy, Sarah N; Wandel, Jasmin; Gross, Thomas; Zysset, Philippe K

    2015-06-01

    As our population ages, more individuals suffer from osteoporosis. This disease leads to impaired trabecular architecture and increased fracture risk. It is essential to understand how morphological and mechanical properties of the cancellous bone are related. Morphology-elasticity relationships based on bone volume fraction (BV/TV) and fabric anisotropy explain up to 98% of the variation in elastic properties. Yet, other morphological variables such as individual trabeculae segmentation (ITS) and trabecular bone score (TBS) could improve the stiffness predictions. A total of 743 micro-computed tomography (μCT) reconstructions of cubic trabecular bone samples extracted from femur, radius, vertebrae, and iliac crest were analyzed. Their morphology was assessed via 25 variables and their stiffness tensor (CFE) was computed from six independent load cases using micro finite element (μFE) analyses. Variance inflation factors were calculated to evaluate collinearity between morphological variables and decide upon their inclusion in morphology-elasticity relationships. The statistically admissible morphological variables were included in a multiple linear regression model of the dependent variable CFE. The contribution of each independent variable was evaluated (ANOVA). Our results show that BV/TV is the best determinant of CFE(r(2) adj  = 0.889), especially in combination with fabric anisotropy (r(2) adj  = 0.968). Including the other independent predictors hardly affected the amount of variance explained by the model (r(2) adj  = 0.975). Across all anatomical sites, BV/TV explained 87% of the variance of the bone elastic properties. Fabric anisotropy further described 10% of the bone stiffness, but the improvement in variance explanation by adding other independent factors was marginal (variables do not bring any further contribution. These overall conclusions remain to be confirmed for specific bone diseases and postelastic properties.

  12. Colorimetric determinations of lithium levels in drop-volumes of human plasma for monitoring patients with bipolar mood disorder.

    Science.gov (United States)

    Qassem, M; Hickey, M; Kyriacou, P A

    2016-08-01

    Lithium preparations are considered the most reliable form of mood stabilizing medication for patients with Bipolar disorder. Nevertheless, lithium is a toxic element and its therapeutic range is extremely narrow, with levels of 0.61.0 mEq considered normal, whereas levels above 1.5 mEq are toxic. Thus unfortunately, many patients reach toxic levels that lead to unnecessary complications. It is believed that personal monitoring of blood lithium levels would benefit patients taking lithium medication. Therefore, our aim is to develop a personal lithium blood level analyzer for patients with bipolar mood disorder, and we report here our initial results of a colorimetric-based method used to test drop-volumes of human plasma that had been spiked with lithium. It was possible to validate results with standard flame photometry readings. Applying the Partial Least Squares (PLS) method on preprocessed spectra, therapeutic concentrations of lithium in a single drop can be predicted in a rapid manner, and furthermore, the calibration results were used to select effective wavelengths which were employed as inputs in Multiple Linear Regression (MLR). The simplified algorithms of this would prove useful when developing a personal lithium analyzer. Overall, both calibration methods gave high correlation and small error outputs with a R2= 0.99036 and RMSEC = 0.03778, and R2= 0.994148 and RMSEC= 0.0294404, for PLS and MLR methods, respectively. The results show that the spectrophotometric determination of blood lithium levels can be extended beyond laboratory applications and indicate the capability of this testing principle to be employed in a personal monitoring device. Future work will now focus on the technical development of a miniaturized system for measurement of lithium levels in blood with an acceptable level of accuracy and sensitivity.

  13. Determination of the Surface and Volume Porosity, on the Basis of the Main Fraction of the Polifractional Matrix of Moulding and Core Sands

    Directory of Open Access Journals (Sweden)

    Dańko R.

    2016-12-01

    Full Text Available The aim of the hereby paper is to present the developed model of determining the volume and surface porosity based on the main fraction of polifractional materials, its experimental verification and utilisation for the interpretation of effects accompanying the formation of a moulding sand apparent density, porosity and permeability in the blowing processes of the core and moulds technology.

  14. Red blood cell volume as a predictor of fatal reactions in cattle infected with Theileria parva Katete

    Directory of Open Access Journals (Sweden)

    P. Fandamu

    2007-09-01

    Full Text Available A comparison of mean corpuscular volume (MCV and packed cell volume (PCV was made between cattle undergoing lethal and non-lethal reactions following experimental infections with the apicomplexan protozoa, Theileria parva Katete. This work confirmed that anaemia occurs in infected animals. However, the fall in PCV was steeper in lethal reactions compared to non-lethal reactions. Our results show that animals with initially lower MCV values are more prone to fatal reaction, despite having normal PCV profiles. The study also found that small red blood cells are more likely to be infected with T. parva. These findings suggest that animals with a higher proportion of small red blood cells in circulation will be more likely to succumb to T. parva infections. The potential for using MCV as a predictor of the outcome of infection challenge is discussed.

  15. Are you bleeding? Validation of a machine-learning algorithm for determination of blood volume status: application to remote triage

    National Research Council Canada - National Science Library

    Caroline A. Rickards; Nisarg Vyas; Kathy L. Ryan; Kevin R. Ward; David Andre; Gennifer M. Hurst; Chelsea R. Barrera; Victor A. Convertino

    2014-01-01

    .... The purpose of this study was to test the hypothesis that low-level physiological signals can be used to develop a machine-learning algorithm for tracking changes in central blood volume that will...

  16. A stereological analysis of NPY, POMC, Orexin, GFAP astrocyte, and Iba1 microglia cell number and volume in diet-induced obese male mice.

    Science.gov (United States)

    Lemus, Moyra B; Bayliss, Jacqueline A; Lockie, Sarah H; Santos, Vanessa V; Reichenbach, Alex; Stark, Romana; Andrews, Zane B

    2015-05-01

    The hypothalamic arcuate nucleus (ARC) contains 2 key neural populations, neuropeptide Y (NPY) and proopiomelanocortin (POMC), and, together with orexin neurons in the lateral hypothalamus, plays an integral role in energy homeostasis. However, no studies have examined total neuronal number and volume after high-fat diet (HFD) exposure using sophisticated stereology. We used design-based stereology to estimate NPY and POMC neuronal number and volume, as well as glial fibrillary acidic protein (astrocyte marker) and ionized calcium-binding adapter molecule 1 (microglia marker) cell number in the ARC; as well as orexin neurons in the lateral hypothalamus. Stereological analysis indicated approximately 8000 NPY and approximately 9000 POMC neurons in the ARC, and approximately 7500 orexin neurons in the lateral hypothalamus. HFD exposure did not affect total neuronal number in any population. However, HFD significantly increased average NPY cell volume and affected NPY and POMC cell volume distribution. HFD reduced orexin cell volume but had a bimodal effect on volume distribution with increased cells at relatively small volumes and decreased cells with relatively large volumes. ARC glial fibrillary acidic protein cells increased after 2 months on a HFD, although no significant difference after 6 months on chow diet or HFD was observed. No differences in ARC ionized calcium-binding adapter molecule 1 cell number were observed in any group. Thus, HFD affects ARC NPY or POMC neuronal cell volume number not cell number. Our results demonstrate the importance of stereology to perform robust unbiased analysis of cell number and volume. These data should be an empirical baseline reference to which future studies are compared.

  17. Is the Maxillary Sinus Really Suitable in Sex Determination? A Three-Dimensional Analysis of Maxillary Sinus Volume and Surface Depending on Sex and Dentition.

    Science.gov (United States)

    Möhlhenrich, Stephan Christian; Heussen, Nicole; Peters, Florian; Steiner, Timm; Hölzle, Frank; Modabber, Ali

    2015-11-01

    The morphometric analysis of maxillary sinus was recently presented as a helpful instrument for sex determination. The aim of the present study was to examine the volume and surface of the fully dentate, partial, and complete edentulous maxillary sinus depending on the sex. Computed tomography data from 276 patients were imported in DICOM format via special virtual planning software, and surfaces (mm) and volumes (mm) of maxillary sinuses were measured. In sex-specific comparisons (women vs men), statistically significant differences for the mean maxillary sinus volume and surface were found between fully dentate (volume, 13,267.77 mm vs 16,623.17 mm, P sexes, no differences were only measured for female and male partially dentate fully edentulous sinuses (2 teeth missing) and between partially edentulous sinuses in women and men (1 teeth vs 2 teeth missing). With a corresponding software program, it is possible to analyze the maxillary sinus precisely. The dentition influences the volume and surface of the pneumatic maxillary sinus. Therefore, sex determination is possible by analysis of the maxillary sinus event through the increase in pneumatization.

  18. Therapeutic analysis of high-dose-rate {sup 192}Ir vaginal cuff brachytherapy for endometrial cancer using a cylindrical target volume model and varied cancer cell distributions

    Energy Technology Data Exchange (ETDEWEB)

    Zhang, Hualin, E-mail: hualin.zhang@northwestern.edu; Donnelly, Eric D.; Strauss, Jonathan B. [Department of Radiation Oncology, Robert H. Lurie Comprehensive Cancer Center, Northwestern University Feinberg School of Medicine, Northwestern Memorial Hospital, Chicago, Illinois 60611 (United States); Qi, Yujin [Centre for Medical Radiation Physics, University of Wollongong, Wollongong, NSW 2522 (Australia)

    2016-01-15

    Purpose: To evaluate high-dose-rate (HDR) vaginal cuff brachytherapy (VCBT) in the treatment of endometrial cancer in a cylindrical target volume with either a varied or a constant cancer cell distributions using the linear quadratic (LQ) model. Methods: A Monte Carlo (MC) technique was used to calculate the 3D dose distribution of HDR VCBT over a variety of cylinder diameters and treatment lengths. A treatment planning system (TPS) was used to make plans for the various cylinder diameters, treatment lengths, and prescriptions using the clinical protocol. The dwell times obtained from the TPS were fed into MC. The LQ model was used to evaluate the therapeutic outcome of two brachytherapy regimens prescribed either at 0.5 cm depth (5.5 Gy × 4 fractions) or at the vaginal mucosal surface (8.8 Gy × 4 fractions) for the treatment of endometrial cancer. An experimentally determined endometrial cancer cell distribution, which showed a varied and resembled a half-Gaussian distribution, was used in radiobiology modeling. The equivalent uniform dose (EUD) to cancer cells was calculated for each treatment scenario. The therapeutic ratio (TR) was defined by comparing VCBT with a uniform dose radiotherapy plan in term of normal cell survival at the same level of cancer cell killing. Calculations of clinical impact were run twice assuming two different types of cancer cell density distributions in the cylindrical target volume: (1) a half-Gaussian or (2) a uniform distribution. Results: EUDs were weakly dependent on cylinder size, treatment length, and the prescription depth, but strongly dependent on the cancer cell distribution. TRs were strongly dependent on the cylinder size, treatment length, types of the cancer cell distributions, and the sensitivity of normal tissue. With a half-Gaussian distribution of cancer cells which populated at the vaginal mucosa the most, the EUDs were between 6.9 Gy × 4 and 7.8 Gy × 4, the TRs were in the range from (5.0){sup 4} to (13

  19. Therapeutic analysis of high-dose-rate (192)Ir vaginal cuff brachytherapy for endometrial cancer using a cylindrical target volume model and varied cancer cell distributions.

    Science.gov (United States)

    Zhang, Hualin; Donnelly, Eric D; Strauss, Jonathan B; Qi, Yujin

    2016-01-01

    To evaluate high-dose-rate (HDR) vaginal cuff brachytherapy (VCBT) in the treatment of endometrial cancer in a cylindrical target volume with either a varied or a constant cancer cell distributions using the linear quadratic (LQ) model. A Monte Carlo (MC) technique was used to calculate the 3D dose distribution of HDR VCBT over a variety of cylinder diameters and treatment lengths. A treatment planning system (TPS) was used to make plans for the various cylinder diameters, treatment lengths, and prescriptions using the clinical protocol. The dwell times obtained from the TPS were fed into MC. The LQ model was used to evaluate the therapeutic outcome of two brachytherapy regimens prescribed either at 0.5 cm depth (5.5 Gy × 4 fractions) or at the vaginal mucosal surface (8.8 Gy × 4 fractions) for the treatment of endometrial cancer. An experimentally determined endometrial cancer cell distribution, which showed a varied and resembled a half-Gaussian distribution, was used in radiobiology modeling. The equivalent uniform dose (EUD) to cancer cells was calculated for each treatment scenario. The therapeutic ratio (TR) was defined by comparing VCBT with a uniform dose radiotherapy plan in term of normal cell survival at the same level of cancer cell killing. Calculations of clinical impact were run twice assuming two different types of cancer cell density distributions in the cylindrical target volume: (1) a half-Gaussian or (2) a uniform distribution. EUDs were weakly dependent on cylinder size, treatment length, and the prescription depth, but strongly dependent on the cancer cell distribution. TRs were strongly dependent on the cylinder size, treatment length, types of the cancer cell distributions, and the sensitivity of normal tissue. With a half-Gaussian distribution of cancer cells which populated at the vaginal mucosa the most, the EUDs were between 6.9 Gy × 4 and 7.8 Gy × 4, the TRs were in the range from (5.0)(4) to (13.4)(4) for the radiosensitive normal

  20. Therapeutic analysis of high-dose-rate 192Ir vaginal cuff brachytherapy for endometrial cancer using a cylindrical target volume model and varied cancer cell distributions

    Science.gov (United States)

    Zhang, Hualin; Donnelly, Eric D.; Strauss, Jonathan B.; Qi, Yujin

    2016-01-01

    Purpose: To evaluate high-dose-rate (HDR) vaginal cuff brachytherapy (VCBT) in the treatment of endometrial cancer in a cylindrical target volume with either a varied or a constant cancer cell distributions using the linear quadratic (LQ) model. Methods: A Monte Carlo (MC) technique was used to calculate the 3D dose distribution of HDR VCBT over a variety of cylinder diameters and treatment lengths. A treatment planning system (TPS) was used to make plans for the various cylinder diameters, treatment lengths, and prescriptions using the clinical protocol. The dwell times obtained from the TPS were fed into MC. The LQ model was used to evaluate the therapeutic outcome of two brachytherapy regimens prescribed either at 0.5 cm depth (5.5 Gy × 4 fractions) or at the vaginal mucosal surface (8.8 Gy × 4 fractions) for the treatment of endometrial cancer. An experimentally determined endometrial cancer cell distribution, which showed a varied and resembled a half-Gaussian distribution, was used in radiobiology modeling. The equivalent uniform dose (EUD) to cancer cells was calculated for each treatment scenario. The therapeutic ratio (TR) was defined by comparing VCBT with a uniform dose radiotherapy plan in term of normal cell survival at the same level of cancer cell killing. Calculations of clinical impact were run twice assuming two different types of cancer cell density distributions in the cylindrical target volume: (1) a half-Gaussian or (2) a uniform distribution. Results: EUDs were weakly dependent on cylinder size, treatment length, and the prescription depth, but strongly dependent on the cancer cell distribution. TRs were strongly dependent on the cylinder size, treatment length, types of the cancer cell distributions, and the sensitivity of normal tissue. With a half-Gaussian distribution of cancer cells which populated at the vaginal mucosa the most, the EUDs were between 6.9 Gy × 4 and 7.8 Gy × 4, the TRs were in the range from (5.0)4 to (13.4)4 for the

  1. An Approach to Determining the Optimal Cell Number of Manufacturing Cell Formation

    Directory of Open Access Journals (Sweden)

    Jianwei Wang

    2013-04-01

    Full Text Available An approach to determining the optimal cell number of manufacturing cell formation is presented. Firstly, the difference of weighting exponent, cluster center and metrics how to have an impact upon the clustering results and membership function are studied. Secondly, a method to determine the optimal m value is given. Two-order partial derivative of the objective function for FCM is calculated, and the variational weighting exponent m is obtained that can prevent the parameter from being the unique value and play an important role in the process of fuzzy clustering. Moreover, in order to avoid a single validity index can not assess correctly, partition coefficient (PC, classification entropy (CE, Fukuyama and Sugeno (FS and Xie and Beni (XB are considered as multi-performance indexes to evaluate the cluster validity, and then an optimal number c is chosen based on these validity measures. Finally, test exampls are given to illustrate the validity of the proposed approach.

  2. Cell Volume Effect on the Ferroelectric Stability of Perovskite Oxides PbTiO3 and BaTiO3 from First Principle Calculation

    Institute of Scientific and Technical Information of China (English)

    王渊旭; 王春雷

    2003-01-01

    Electronic structure of ferroelectric PbTiO3 and BaTiO3 is calculated by the full potential linearized augmented plane wave method. The total energy as a function of the displacement of Ti-cation is obtained for PbTiO3 and BaTiO3 at different cell volumes. At experimental cell volume, Ti-displacement lowers the total energy and the ferroelectricity is stable. When the cell volume is reduced to 90%, total energy is increased with Ti-displacement and ferroelectricity will disappear. The cell volume effect is also confirmed by comparison of the density of states of Ti and O at different cell volumes.

  3. Myeloid dendritic cells frequencies are increased in children with autism spectrum disorder and associated with amygdala volume and repetitive behaviors.

    Science.gov (United States)

    Breece, Elizabeth; Paciotti, Brian; Nordahl, Christine Wu; Ozonoff, Sally; Van de Water, Judy A; Rogers, Sally J; Amaral, David; Ashwood, Paul

    2013-07-01

    The pathophysiology of autism spectrum disorder (ASD) is not yet known; however, studies suggest that dysfunction of the immune system affects many children with ASD. Increasing evidence points to dysfunction of the innate immune system including activation of microglia and perivascular macrophages, increases in inflammatory cytokines/chemokines in brain tissue and CSF, and abnormal peripheral monocyte cell function. Dendritic cells are major players in innate immunity and have important functions in the phagocytosis of pathogens or debris, antigen presentation, activation of naïve T cells, induction of tolerance and cytokine/chemokine production. In this study, we assessed circulating frequencies of myeloid dendritic cells (defined as Lin-1(-)BDCA1(+)CD11c(+) and Lin-1(-)BDCA3(+)CD123(-)) and plasmacytoid dendritic cells (Lin-1(-)BDCA2(+)CD123(+) or Lin-1(-)BDCA4(+) CD11c(-)) in 57 children with ASD, and 29 typically developing controls of the same age, all of who were enrolled as part of the Autism Phenome Project (APP). The frequencies of dendritic cells and associations with behavioral assessment and MRI measurements of amygdala volume were compared in the same participants. The frequencies of myeloid dendritic cells were significantly increased in children with ASD compared to typically developing controls (pfrequencies of myeloid dendritic cells were positively associated with abnormal right and left amygdala enlargement, severity of gastrointestinal symptoms and increased repetitive behaviors. The frequencies of plasmacytoid dendritic cells were also associated with amygdala volumes as well as developmental regression in children with ASD. Dendritic cells play key roles in modulating immune responses and differences in frequencies or functions of these cells may result in immune dysfunction in children with ASD. These data further implicate innate immune cells in the complex pathophysiology of ASD.

  4. Cancer Detection via Determination of Fractal Cell Dimension

    CERN Document Server

    Bauer, W; Bauer, Wolfgang; Mackenzie, Charles D.

    1995-01-01

    We utilize the fractal dimension of the perimeter surface of cell sections as a new observable to characterize cells of different types. We propose that it is possible to distinguish cancerous from healthy cells with the aid of this new approach. As a first application we show that it is possible to perform this distinction between patients with hairy-cell lymphocytic leukemia and those with normal blood lymphocytes.

  5. Stereological estimation of nuclear volume and other quantitative histopathological parameters in the prognostic evaluation of supraglottic laryngeal squamous cell carcinoma

    DEFF Research Database (Denmark)

    Sørensen, Flemming Brandt; Bennedbaek, O; Pilgaard, J;

    1989-01-01

    observers of the latter was poor in the material which consisted of 35 biopsy specimens. Unbiased estimates of nuclear Vv were on the average 385 microns3 (CV = 0.44), with more than 90% of the associated variance attributable to differences in nuclear Vv among individual lesions. Nuclear Vv was positively......The aim of this study was to investigate various approaches to the grading of malignancy in pre-treatment biopsies from patients with supraglottic laryngeal squamous cell carcinoma. The prospects of objective malignancy grading based on stereological estimation of the volume-weighted mean nuclear...... volume, nuclear Vv, and nuclear volume fraction, Vv(nuc/tis), along with morphometrical 2-dimensional estimation of nuclear density index, NI, and mitotic activity index, MI, were investigated and compared with the current morphological, multifactorial grading system. The reproducibility among two...

  6. Induction of Inducible Nitric Oxide Synthase by Lipopolysaccharide and the Influences of Cell Volume Changes, Stress Hormones and Oxidative Stress on Nitric Oxide Efflux from the Perfused Liver of Air-Breathing Catfish, Heteropneustes fossilis.

    Science.gov (United States)

    Choudhury, Mahua G; Saha, Nirmalendu

    2016-01-01

    The air-breathing singhi catfish (Heteropneustes fossilis) is frequently being challenged by bacterial contaminants, and different environmental insults like osmotic, hyper-ammonia, dehydration and oxidative stresses in its natural habitats throughout the year. The main objectives of the present investigation were to determine (a) the possible induction of inducible nitric oxide synthase (iNOS) gene with enhanced production of nitric oxide (NO) by intra-peritoneal injection of lipopolysaccharide (LPS) (a bacterial endotoxin), and (b) to determine the effects of hepatic cell volume changes due to anisotonicity or by infusion of certain metabolites, stress hormones and by induction of oxidative stress on production of NO from the iNOS-induced perfused liver of singhi catfish. Intra-peritoneal injection of LPS led to induction of iNOS gene and localized tissue specific expression of iNOS enzyme with more production and accumulation of NO in different tissues of singhi catfish. Further, changes of hydration status/cell volume, caused either by anisotonicity or by infusion of certain metabolites such as glutamine plus glycine and adenosine, affected the NO production from the perfused liver of iNOS-induced singhi catfish. In general, increase of hydration status/cell swelling due to hypotonicity caused decrease, and decrease of hydration status/cell shrinkage due to hypertonicity caused increase of NO efflux from the perfused liver, thus suggesting that changes in hydration status/cell volume of hepatic cells serve as a potent modulator for regulating the NO production. Significant increase of NO efflux from the perfused liver was also observed while infusing the liver with stress hormones like epinephrine and norepinephrine, accompanied with decrease of hydration status/cell volume of hepatic cells. Further, oxidative stress, caused due to infusion of t-butyl hydroperoxide and hydrogen peroxide separately, in the perfused liver of singhi catfish, resulted in

  7. Detection of atomic scale changes in the free volume void size of three-dimensional colorectal cancer cell culture using positron annihilation lifetime spectroscopy.

    Science.gov (United States)

    Axpe, Eneko; Lopez-Euba, Tamara; Castellanos-Rubio, Ainara; Merida, David; Garcia, Jose Angel; Plaza-Izurieta, Leticia; Fernandez-Jimenez, Nora; Plazaola, Fernando; Bilbao, Jose Ramon

    2014-01-01

    Positron annihilation lifetime spectroscopy (PALS) provides a direct measurement of the free volume void sizes in polymers and biological systems. This free volume is critical in explaining and understanding physical and mechanical properties of polymers. Moreover, PALS has been recently proposed as a potential tool in detecting cancer at early stages, probing the differences in the subnanometer scale free volume voids between cancerous/healthy skin samples of the same patient. Despite several investigations on free volume in complex cancerous tissues, no positron annihilation studies of living cancer cell cultures have been reported. We demonstrate that PALS can be applied to the study in human living 3D cell cultures. The technique is also capable to detect atomic scale changes in the size of the free volume voids due to the biological responses to TGF-β. PALS may be developed to characterize the effect of different culture conditions in the free volume voids of cells grown in vitro.

  8. Comparison of two flow-based imaging methods to measure individual red blood cell area and volume.

    Science.gov (United States)

    Tomaiuolo, Giovanna; Rossi, Domenico; Caserta, Sergio; Cesarelli, Mario; Guido, Stefano

    2012-12-01

    The red blood cells (RBCs) population is characterized by a high heterogeneity in membrane area, cellular volume, and mechanical properties, mainly due to the variety of mechanical and chemical stresses that a red cell undergoes in its entire life span. Here, we provide the first simultaneous area and volume measurements of RBCs flowing in microcapillaries, by using high-speed video microscopy imaging and quantitative data processing based on image analysis techniques. Both confined and unbounded flow conditions (depending on the relative size of RBCs and microcapillary diameter) are investigated. The results are compared with micropipette experiments from the literature and data from Coulter counter routine clinical blood tests. Good agreement is found for RBC volume, especially in the case of confined flow conditions. Surface area measurements, which are lacking in the routine clinical test, are of special interest being a potential diagnostic parameter of altered cell deformability and aggregability. Overall, our results provide a novel flow methodology suitable for high-throughput measurements of RBC geometrical parameters, allowing one to overcome the limits of classical static methods, such as micropipette aspiration, which are not suitable for handling a large number of cells.

  9. Sex determination using discriminant analysis of upper and lower extremity bones: New approach using the volume and surface area of digital model.

    Science.gov (United States)

    Lee, U-Young; Kim, In-Beom; Kwak, Dai-Soon

    2015-08-01

    This study used 110 CT images taken from donated Korean cadavers to create 3-D models of the following upper and lower limb bones: the clavicle, scapula, humerus, radius, ulna, hip bone (os coxa), femur, patella (knee cap), tibia, talus, and calcaneus. In addition, the bone volume and surface area were calculated to determine sex differences using discriminant analysis. Significant sex differences were found in all bones with respect to volume and surface area (phip bone>tibia>humerus>scapula), although the order of surface area was different. The largest surface area in men was the femur and in women was the hip bone (psex determination (94%). When using the surface area of multiple bones, the maximum accuracy (99.4%) was achieved. The equation was as follows: (discriminant equation of surface area; femalesex determination.

  10. Using Ray Tracing to Evaluate the Performance of Several Methods for Determining the Ground Range and Refractive Index of Ionospheric Scattering Volumes

    Science.gov (United States)

    Greenwald, R. A.; Frissell, N. A.; de Larquier, S.

    2016-12-01

    In this paper, we evaluate the performance of three methods used by HF radars in the SuperDARN network for determining the ground ranges to ionospheric scattering volumes. Each method uses somewhat different approaches, but the same equivalent-path analysis. We also show that Snell's Law can be added to this analysis to determine the refractive index of each scattering volume and thereby correct Doppler velocity measurements for ionospheric refraction. Two of these methods make their predictions using the group range to the scattering volume and a virtual height model, while the third method uses the group range and the elevation angle each backscattered return. The effectiveness of each of these methods is evaluated using ray tracing analyses through the International Reference Ionosphere. Ray tracings analysis provides determinations of the initial elevation angle, group range, group range, and refractive index of each ionospheric volume that backscatters signals to the radar. The initial or final elevation angle and the group range are used as inputs to the geolocation methods and the ground range and refractive index serve as reference data against which the predictions of the geolocation methods can be evaluated. We find that the methods using virtual height models actually change the initial elevation angle determined from ray tracing to a different elevation angle that is consistent with the virtual height model. Due to this change, predictions of the ground range and refractive index of scattering volumes located with virtual-height models are rarely consistent with the predictions obtained from ray tracing. In contrast, the geolocation method that uses the group range and initial or final elevation angle yields predictions that are in good agreement with ray tracing. Modifications to the equivalent-path analysis are required to obtain consistent predictions of the ground range and refractive index of backscatter from the topside F-layer.

  11. [Relationship between testis volume and types of spermatogenic cells from testicular biopsy in patients with azoospermia or cryptozoospermia].

    Science.gov (United States)

    Liu, Xing-Zhang; Tang, Yun-Ge; Liu, Huang; Tang, Li-Xin; Wen, Ren-Qian

    2010-01-01

    To study the relationship between the testis volume and types of spermatogenic cells derived from testicular biopsy in patients with azoospermia or cryptozoospermia. We collected testicular pathological biopsies from 492 infertile patients with azoospermia or cryptozoospermia reported in our hospital, classified them according to the testicular histological classification methods in WHO Manual for Standardized Investigation, Diagnosis and Management of the Infertile Male, and analyzed the relationship of the testis volume with the results of semen analyses and testicular histology. Of the 492 cases, 90.5% (445/492) were azoospermia and 9.5% (47/492) cryptozoospermia; mature spermatozoa were present in the seminiferous tubules in 17.9% (88/492) but absent in 42.9% (211/492), and Sertoli cell-only syndrome indicated in 39.2% (193/492); the testis volume was biopsy, and it may exist even if the testis volume is significantly below the reference value. The indications for testicular biopsy should not be improperly expanded. The WHO testicular histological classification methods have provided a convenient and effective guidance for further clinical examinations and establishment of a protocol.

  12. Artifacts in conventional computed tomography (CT) and free breathing four-dimensional CT induce uncertainty in gross tumor volume determination

    DEFF Research Database (Denmark)

    Persson, Gitte Fredberg; Nygaard, Ditte Eklund; Af Rosenschöld, Per Munck;

    2011-01-01

    PURPOSE: Artifacts impacting the imaged tumor volume can be seen in conventional three-dimensional CT (3DCT) scans for planning of lung cancer radiotherapy but can be reduced with the use of respiration-correlated imaging, i.e., 4DCT or breathhold CT (BHCT) scans. The aim of this study...... was to compare delineated gross tumor volume (GTV) sizes in 3DCT, 4DCT, and BHCT scans of patients with lung tumors. METHODS AND MATERIALS: A total of 36 patients with 46 tumors referred for stereotactic radiotherapy of lung tumors were included. All patients underwent positron emission tomography (PET)/CT, 4DCT......, and BHCT scans. GTVs in all CT scans of individual patients were delineated during one session by a single physician to minimize systematic delineation uncertainty. The GTV size from the BHCT was considered the closest to true tumor volume and was chosen as the reference. The reference GTV size...

  13. Sendai virus utilizes specific sialyloligosaccharides as host cell receptor determinants.

    OpenAIRE

    1980-01-01

    Purified sialyltransferases (CMP-N-acetyl-neuraminate:D-galactosyl-glycoprotein N-acetylneuraminyl-transferase, EC 2.4.99.1) in conjunction with neuraminidase (acylneuraminyl hydrolase, EC 3.2.1.18) were used to produce cell surface sialyloligosaccharides of defined sequence to investigate their role in paramyxovirus infection of host cells. Infection of Madin-Darby bovine kidney cells by Sendai virus was monitored by hemagglutination titer of the virus produced and by changes in morphologica...

  14. A transient, HEX-Z nodal code corrected by discontinuity factors. Volume 2, Monte Carlo determination of discontinuity factors

    Energy Technology Data Exchange (ETDEWEB)

    Mohamed, A.S.G.; Henry, A.F.

    1993-12-31

    This document constitutes Volume 2 of the Final Report of a three- year study supported by the special Research Grant Program for Nuclear Energy Research set up by the US Department of Energy. Since the material content is entirely distinct from that of Volume 1, the present report is written as a stand-along document. The original motivation for the overall research effort was to provide a fast and accurate computer program for the analysis of transients in heavy water or graphite-moderated reactors being considered as candidates for the New Production Reactor.

  15. Steady-state solutions of cell volume in a cardiac myocyte model elaborated for membrane excitation, ion homeostasis and Ca2+ dynamics.

    Science.gov (United States)

    Cha, Chae Young; Noma, Akinori

    2012-08-21

    The cell volume continuously changes in response to varying physiological conditions, and mechanisms underlying volume regulation have been investigated in both experimental and theoretical studies. Here, general formulations concerning cell volume change are presented in the context of developing a comprehensive cell model which takes Ca(2+) dynamics into account. Explicit formulas for charge conservation and steady-state volumes of the cytosol and endoplasmic reticulum (ER) are derived in terms of membrane potential, amount of ions, Ca(2+)-bound buffer molecules, and initial cellular conditions. The formulations were applied to a ventricular myocyte model which has plasma-membrane Ca(2+) currents with dynamic gating mechanisms, Ca(2+)-buffering reactions with diffusive and non-diffusive buffer proteins, and Ca(2+) uptake into or release from the sarcoplasmic reticulum (SR) accompanied by compensatory cationic or anionic currents through the SR membrane. Time-dependent volume changes in cardiac myocytes induced by varying extracellular osmolarity or by action potential generation were successfully simulated by the novel formulations. Through application of bifurcation analysis, the existence and uniqueness of steady-state solutions of the cell volume were validated, and contributions of individual ion channels and transporters to the steady-state volume were systematically analyzed. The new formulas are consistent with previous fundamental theory derived from simple models of minimum compositions. The new formulations may be useful for examination of the relationship between cell function and volume change in other cell types.

  16. Multimodality imaging with CT, MR and FDG-PET for radiotherapy target volume delineation in oropharyngeal squamous cell carcinoma

    OpenAIRE

    Bird, David; Scarsbrook, Andrew F.; Sykes, Jonathan; Ramasamy, Satiavani; Subesinghe, Manil; Carey, Brendan; Wilson, Daniel J.; Roberts, Neil; McDermott, Gary; KARAKAYA, Ebru; BAYMAN, Evrim; Sen, Mehmet; Speight, Richard; Prestwich, Robin J. D.

    2015-01-01

    Background This study aimed to quantify the variation in oropharyngeal squamous cell carcinoma gross tumour volume (GTV) delineation between CT, MR and FDG PET-CT imaging. Methods A prospective, single centre, pilot study was undertaken where 11 patients with locally advanced oropharyngeal cancers (2 tonsil, 9 base of tongue primaries) underwent pre-treatment, contrast enhanced, FDG PET-CT and MR imaging, all performed in a radiotherapy treatment mask. CT, MR and CT-MR GTVs were contoured by ...

  17. Tolerance and dose-volume relationship of intrathoracic stomach irradiation after esophagectomy for patients with thoracic esophageal squamous cell carcinoma

    OpenAIRE

    Liu, Qi; Cai, Xu-Wei; Fu, Xiao-Long; Chen, Jun-Chao; Xiang, Jia-Qing

    2015-01-01

    Purpose To identify the tolerance of radiation with a high prescribed dose and predictors for the development of intrathoracic stomach toxicity in patients with thoracic esophageal squamous cell carcinoma (SCC) after esophagectomy followed by gastric conduit reconstruction. Methods and Materials From 2011 to 2013, 105 patients after esophagectomy were treated with postoperative radiotherapy. The intrathoracic stomach was outlined with the calculation of a dose-volume histogram (DVH) for the i...

  18. Experience with a Fourier method for determining the extracellular potential fields of excitable cells with cylindrical geometry.

    Science.gov (United States)

    Clark, J W; Greco, E C; Harman, T L

    1978-11-01

    In this chapter, well-known solutions that utilize a Fourier transform method for determining the extracellular, volume-conductor potential distribution surrounding elongated excitable cells of cylindrical geometry are reformulated as a discrete Fourier transform (DFT) problem, which subsequently permits the volume-conductor problem to be viewed as an equivalent linear-filtering problem. This DFT formulation is fast and computationally efficient. In addition, it lends itself to the application of some rather well-known techniques in linear systems theory (e.g., the DFT for convolution and least mean-square (Wiener) filtering for optimal prediction of a signal in random noise). Two specific examples are employed to demonstrate the utility of this discrete Fourier method: (1) the single, isolated, active nerve fiber in an essentially infinite volume conductor and (2) the isolated, active nerve trunk in a similar type of extracellular medium. In each of these, our DFT method is employed to obtain both the classical "forward" and "inverse" potential solutions for each volume conductor problem. In the case where the single, active nerve fiber is the bioelectric source in the volume conductor, simulated action-potential data from an invertebrate giant axon is utilized, and potentials at various points in the extracellular medium are calculated. The calculated potential distributions in axial distance z, at various radial distances r, are consistent with well-known experimental fact. When the active nerve trunk acts as the bioelectric source, the DFT method provides calculated potential distributions that are fairly consistent with experimental data under a variety of experimental conditions. For example, in these experiments, a special, isolated frog spinal cord preparation is used that permits separate or combined stimulation of the motor and sensory nerve fiber components of the attached sciatic nerve trunk. By manipulating the stimulus intensity applied to the motor

  19. Determination of individual cell Michaelis-Menten constants.

    Science.gov (United States)

    Sunray, Merav; Zurgil, Naomi; Shafran, Yana; Deutsch, Mordechai

    2002-01-01

    A novel methodology for the measurement and analysis of apparent K(M) (Michaelis-Menten constant) and V(MAX) values of individual cells is suggested. It is based on a mathematical model that considers substrate influx into the cell, its intracellular enzymatic hydrolysis, and the product efflux. The mathematical formulation was approximated linearly in order to analyze intracellular substrate conversion characteristics via Michaelis-Menten theory. Utilizing static cytometry, the time dependence of the fluorescence intensity [FI(t)] emitted from prelocalized and defined FDA stained cells was recorded. This required frequent periodical measurements of the same cells, which are sequentially exposed to various fluorogenic substrate concentrations. Model simulations correlated with experimental results. Differences in distributions of individual K(M) and V(MAX) values of cells incubated with and without PHA were evident. Average K(M) and V(MAX) values of PHA-stimulated cells increased by 99% and 540%, respectively. This study may provide a tool for assessing intracellular enzymatic activity in individual intact cells under defined physiologic conditions. This may open new vistas in various areas, giving answers to critical questions arising in the field of cell and developmental biology, immunology, oncology, and pharmacology. Copyright 2001 Wiley-Liss, Inc.

  20. Quantitative sodium MRI of the human brain at 9.4 T provides assessment of tissue sodium concentration and cell volume fraction during normal aging.

    Science.gov (United States)

    Thulborn, Keith; Lui, Elaine; Guntin, Jonathan; Jamil, Saad; Sun, Ziqi; Claiborne, Theodore C; Atkinson, Ian C

    2016-02-01

    Sodium ion homeostasis is a fundamental property of viable tissue, allowing the tissue sodium concentration to be modeled as the tissue cell volume fraction. The modern neuropathology literature using ex vivo tissue from selected brain regions indicates that human brain cell density remains constant during normal aging and attributes the volume loss that occurs with advancing age to changes in neuronal size and dendritic arborization. Quantitative sodium MRI performed with the enhanced sensitivity of ultrahigh-field 9.4 T has been used to investigate tissue cell volume fraction during normal aging. This cross-sectional study (n = 49; 21-80 years) finds that the in vivo tissue cell volume fraction remains constant in all regions of the brain with advancing age in individuals who remain cognitively normal, extending the ex vivo literature reporting constant neuronal cell density across the normal adult age range. Cell volume fraction, as measured by quantitative sodium MRI, is decreased in diseases of cell loss, such as stroke, on a time scale of minutes to hours, and in response to treatment of brain tumors on a time scale of days to weeks. Neurodegenerative diseases often have prodromal periods of decades in which regional neuronal cell loss occurs prior to clinical presentation. If tissue cell volume fraction can detect such early pathology, this quantitative parameter may permit the objective measurement of preclinical disease progression. This current study in cognitively normal aging individuals provides the basis for the pursuance of investigations directed towards such neurodegenerative diseases.

  1. Oyster's cells regulatory volume decrease: A new tool for evaluating the toxicity of low concentration hydrocarbons in marine waters.

    Science.gov (United States)

    Ben Naceur, Chiraz; Maxime, Valérie; Ben Mansour, Hedi; Le Tilly, Véronique; Sire, Olivier

    2016-11-01

    Human activities require fossil fuels for transport and energy, a substantial part of which can accidentally or voluntarily (oil spillage) flow to the marine environment and cause adverse effects in human and ecosystems' health. This experiment was designed to estimate the suitability of an original cellular biomarker to early quantify the biological risk associated to hydrocarbons pollutants in seawater. Oocytes and hepatopancreas cells, isolated from oyster (Crassostrea gigas), were tested for their capacity to regulate their volume following a hypo-osmotic challenge. Cell volumes were estimated from cell images recorded at regular time intervals during a 90min-period. When exposed to diluted seawater (osmolalities from 895 to 712mosmkg(-1)), both cell types first swell and then undergo a shrinkage known as Regulatory Volume Decrease (RVD). This process is inversely proportional to the magnitude of the osmotic shock and is best fitted using a first-order exponential decay model. The Recovered Volume Factor (RVF) calculated from this model appears to be an accurate tool to compare cells responses. As shown by an about 50% decrease in RVF, the RVD process was significantly inhibited in cells sampled from oysters previously exposed to a low concentration of diesel oil (8.4mgL(-1) during 24h). This toxic effect was interpreted as a decreased permeability of the cell membranes resulting from an alteration of their lipidic structure by diesel oil compounds. In contrast, the previous contact of oysters with diesel did not induce any rise in the gills glutathione S-transferase specific activity. Therefore, this work demonstrates that the study of the RVD process of cells selected from sentinel animal species could be an alternative bioassay for the monitoring of hydrocarbons and probably, of various chemicals in the environment liable to alter the cellular regulations. Especially, given the high sensitivity of this biomarker compared with a proven one, it could become a

  2. Definition of molecular determinants of prostate cancer cell bone extravasation.

    Science.gov (United States)

    Barthel, Steven R; Hays, Danielle L; Yazawa, Erika M; Opperman, Matthew; Walley, Kempland C; Nimrichter, Leonardo; Burdick, Monica M; Gillard, Bryan M; Moser, Michael T; Pantel, Klaus; Foster, Barbara A; Pienta, Kenneth J; Dimitroff, Charles J

    2013-01-15

    Advanced prostate cancer commonly metastasizes to bone, but transit of malignant cells across the bone marrow endothelium (BMEC) remains a poorly understood step in metastasis. Prostate cancer cells roll on E-selectin(+) BMEC through E-selectin ligand-binding interactions under shear flow, and prostate cancer cells exhibit firm adhesion to BMEC via β1, β4, and αVβ3 integrins in static assays. However, whether these discrete prostate cancer cell-BMEC adhesive contacts culminate in cooperative, step-wise transendothelial migration into bone is not known. Here, we describe how metastatic prostate cancer cells breach BMEC monolayers in a step-wise fashion under physiologic hemodynamic flow. Prostate cancer cells tethered and rolled on BMEC and then firmly adhered to and traversed BMEC via sequential dependence on E-selectin ligands and β1 and αVβ3 integrins. Expression analysis in human metastatic prostate cancer tissue revealed that β1 was markedly upregulated compared with expression of other β subunits. Prostate cancer cell breaching was regulated by Rac1 and Rap1 GTPases and, notably, did not require exogenous chemokines as β1, αVβ3, Rac1, and Rap1 were constitutively active. In homing studies, prostate cancer cell trafficking to murine femurs was dependent on E-selectin ligand, β1 integrin, and Rac1. Moreover, eliminating E-selectin ligand-synthesizing α1,3 fucosyltransferases in transgenic adenoma of mouse prostate mice dramatically reduced prostate cancer incidence. These results unify the requirement for E-selectin ligands, α1,3 fucosyltransferases, β1 and αVβ3 integrins, and Rac/Rap1 GTPases in mediating prostate cancer cell homing and entry into bone and offer new insight into the role of α1,3 fucosylation in prostate cancer development.

  3. Effects of serum of the rats ventilated with high tidal volume on endothelial cell permeability and therapeutic effects of ulinastatin

    Institute of Scientific and Technical Information of China (English)

    HUO Guo-dong; CAI Shao-xi; CHEN Bo; CHEN Ying-hua

    2006-01-01

    Background With the widespread use of ventilators in treating critically ill patients, the morbidity of ventilator-induced lung injury (VILI) is increasing accordingly. VILI is characterized by a considerable increase in microvascular leakiness and activation of inflammatory processes. In this study we investigated the effects of inflammatory mediators in VILI rat serum on endothelial cytoskeleton and monolayer cellular permeability, as well as the therapeutic effect of ulinastatin, to explore the pathogenesis and the relationship between biotrauma and lung oedema induced by VILI.Methods Thirty healthy male Sprague-Dawley rats were randomly divided into three groups: group A (normal tidal volume ventilation), group B (high tidal volume ventilation) and group C (high tidal volume ventilation plus ulinastatin). The serum of each rat after ventilation was added to endothelial cell line ECV-304 medium for two hours to observe the effects of serum and/or ulinastatin on endothelial fibrous actin and permeability. Results Compared to rats ventilated with normal tidal volume, serum of rats ventilated with high tidal volume caused a striking reorganization of actin cytoskeleton with a weakening of fluorescent intensity at the peripheral filament bands and formation of the long and thick stress fibres in the centre resulting in endothelial contraction and higher permeability. Prior treatment with ulinastatin lessened the above changes significantly. The changes of permeability coefficient of endothelial permeability after group A, B or C rats serum stimulation were (6.95 ±1.66)%, (27.50±7.77)% and (17.71±4.66)% respectively with statistically significant differences (P<0.05)among the three groups.Conclusions The proinflammatory mediators in the serum of the rats given high tidal volume ventilation increases endothelial permeability by reorganizing actin cytoskeleton, and pretreatment with ulinastatin lessens the permeability by inhibiting of proinflammatory mediators.

  4. Prenatal infection decreases calbindin, decreases Purkinje cell volume and density and produces long-term motor deficits in Sprague-Dawley rats.

    Science.gov (United States)

    Wallace, K; Veerisetty, S; Paul, I; May, W; Miguel-Hidalgo, J J; Bennett, W

    2010-01-01

    The cerebellum is involved in the control of motor functions with Purkinje cells serving as the only output from the cerebellum. Purkinje cells are important targets for toxic substances and are vulnerable to prenatal insults. Intrauterine infection (IUI) has been shown to selectively target the developing cerebral white matter through lesioning, necrosis and inflammatory cytokine activation. Developmental and cognitive delays have been associated with animal models of IUI. The aim of this study was to determine if IUI leads to damage to Purkinje cells in the developing cerebellum and if any damage is associated with decreases in calbindin and motor behaviors in surviving pups. Pregnant rats were injected with Escherichia coli (1 × 10⁵ colony-forming units) or sterile saline at gestational day 17. Beginning at postnatal day (PND) 2, the pups were subjected to a series of developmental tests to examine developmental milestones. At PND 16, some pups were sacrificed and their brains extracted and processed for histology or protein studies. Hematoxylin and eosin (HE) staining was done to examine the general morphology of the Purkinje cells and to examine Purkinje cell density, area and volume. Calbindin expression was examined in the cerebellum via immunohistochemistry and Western blot techniques. The remaining rat pups were used to examine motor coordination and balance on a rotating rotarod at the prepubertal and adult ages. Prenatal E. coli injection did not significantly change birth weight or delivery time, but did delay surface righting and negative geotaxis in pups. Pups in the E. coli group also had a decrease in the number of Purkinje cells, as well as a decrease in Purkinje cell density and volume. HE staining demonstrated a change in Purkinje cell morphology. Calbindin expression was decreased in rats from the E. coli group as well. Locomotor tests indicated that while there were no significant changes in gross motor activity, motor coordination and

  5. Automatic cell cloning assay for determining the clonogenic capacity of cancer and cancer stem-like cells.

    Science.gov (United States)

    Fedr, Radek; Pernicová, Zuzana; Slabáková, Eva; Straková, Nicol; Bouchal, Jan; Grepl, Michal; Kozubík, Alois; Souček, Karel

    2013-05-01

    The clonogenic assay is a well-established in vitro method for testing the survival and proliferative capability of cells. It can be used to determine the cytotoxic effects of various treatments including chemotherapeutics and ionizing radiation. However, this approach can also characterize cells with different phenotypes and biological properties, such as stem cells or cancer stem cells. In this study, we implemented a faster and more precise method for assessing the cloning efficiency of cancer stem-like cells that were characterized and separated using a high-speed cell sorter. Cell plating onto a microplate using an automatic cell deposition unit was performed in a single-cell or dilution rank mode by the fluorescence-activated cell sorting method. We tested the new automatic cell-cloning assay (ACCA) on selected cancer cell lines and compared it with the manual approach. The obtained results were also compared with the results of the limiting dilution assay for different cell lines. We applied the ACCA to analyze the cloning capacity of different subpopulations of prostate and colon cancer cells based on the expression of the characteristic markers of stem (CD44 and CD133) and cancer stem cells (TROP-2, CD49f, and CD44). Our results revealed that the novel ACCA is a straightforward approach for determining the clonogenic capacity of cancer stem-like cells identified in both cell lines and patient samples. Copyright © 2013 International Society for Advancement of Cytometry.

  6. 2,3-Dihydroxybenzoic acid attenuates kanamycin-induced volume reduction in mouse utricular type I hair cells

    DEFF Research Database (Denmark)

    Severinsen, Stig Åvall; Kirkegaard, Mette; Nyengaard, Jens Randel

    2006-01-01

    The aminoglycoside kanamycin is a commonly used antibiotic, but unfortunately it is oto- and nephrotoxic in large doses. The negative effects are thought to be due to the formation of free radicals which is why strong antioxidants and iron chelators like 2,3-dihydroxybenzoic acid (DHB) are of great...... interest. This study estimates cellular quantitative changes in the utricular macula of mice following systemic treatment with kanamycin alone or in combination with DHB. The animals were injected with either saline, kanamycin or kanamycin+DHB for 15 days and perfusion fixed three weeks after last...... macula, hair cell type I and supporting cells decreased significantly in animals injected with kanamycin but not in animals co-treated with DHB. Hair and supporting cell numbers remained unchanged in all three groups. In conclusion, the kanamycin-induced volume reduction of type I hair cells...

  7. Cell fate determination during tooth development and regeneration.

    Science.gov (United States)

    Mitsiadis, Thimios A; Graf, Daniel

    2009-09-01

    Teeth arise from sequential and reciprocal interactions between the oral epithelium and the underlying cranial neural crest-derived mesenchyme. Their formation involves a precisely orchestrated series of molecular and morphogenetic events, and gives us the opportunity to discover and understand the nature of the signals that direct cell fates and patterning. For that reason, it is important to elucidate how signaling factors work together in a defined number of cells to generate the diverse and precise patterned structures of the mature functional teeth. Over the last decade, substantial research efforts have been directed toward elucidating the molecular mechanisms that control cell fate decisions during tooth development. These efforts have contributed toward the increased knowledge on dental stem cells, and observation of the molecular similarities that exist between tooth development and regeneration.

  8. Modular, High-Volume Fuel Cell Leak-Test Suite and Process

    Energy Technology Data Exchange (ETDEWEB)

    Ru Chen; Ian Kaye

    2012-03-12

    Fuel cell stacks are typically hand-assembled and tested. As a result the manufacturing process is labor-intensive and time-consuming. The fluid leakage in fuel cell stacks may reduce fuel cell performance, damage fuel cell stack, or even cause fire and become a safety hazard. Leak check is a critical step in the fuel cell stack manufacturing. The fuel cell industry is in need of fuel cell leak-test processes and equipment that is automatic, robust, and high throughput. The equipment should reduce fuel cell manufacturing cost.

  9. Determination of the volume-specific surface area by using transmission electron tomography for characterization and definition of nanomaterials

    Directory of Open Access Journals (Sweden)

    Francisco Michel

    2011-05-01

    Full Text Available Abstract Background Transmission electron microscopy (TEM remains an important technique to investigate the size, shape and surface characteristics of particles at the nanometer scale. Resulting micrographs are two dimensional projections of objects and their interpretation can be difficult. Recently, electron tomography (ET is increasingly used to reveal the morphology of nanomaterials (NM in 3D. In this study, we examined the feasibility to visualize and measure silica and gold NM in suspension using conventional bright field electron tomography. Results The general morphology of gold and silica NM was visualized in 3D by conventional TEM in bright field mode. In orthoslices of the examined NM the surface features of a NM could be seen and measured without interference of higher or lower lying structures inherent to conventional TEM. Segmentation by isosurface rendering allowed visualizing the 3D information of an electron tomographic reconstruction in greater detail than digital slicing. From the 3D reconstructions, the surface area and the volume of the examined NM could be estimated directly and the volume-specific surface area (VSSA was calculated. The mean VSSA of all examined NM was significantly larger than the threshold of 60 m2/cm3. The high correlation between the measured values of area and volume gold nanoparticles with a known spherical morphology and the areas and volumes calculated from the equivalent circle diameter (ECD of projected nanoparticles (NP indicates that the values measured from electron tomographic reconstructions are valid for these gold particles. Conclusion The characterization and definition of the examined gold and silica NM can benefit from application of conventional bright field electron tomography: the NM can be visualized in 3D, while surface features and the VSSA can be measured.

  10. Dispersed and piled woody residues volumes in coastal Douglas-fir cutblocks determined using high-resolution imagery from a UAV and from ground-based surveys.

    Science.gov (United States)

    Trofymow, J. A.; Gougeon, F.

    2015-12-01

    After forest harvest significant amounts of woody residues are left dispersed on site and some subsequently piled and burned. Quantification of residues is required for estimating C budgets, billable waste, harvest efficiency, bioenergy potential and smoke emissions. Trofymow (et al 2014 CJFR) compared remote sensing methods to ground-based waste and residue survey (WRS) methods for residue piles in 4 cutblocks in the Oyster River (OR) area in coastal BC. Compared to geospatial methods using 15cm orthophotos and LiDAR acquired in 2011 by helicopter, the WRS method underestimated pile wood by 30% to 50% while a USFS volume method overestimated pile wood by 50% if site specific packing ratios were not used. A geospatial method was developed in PCI Geomatica to analyze 2-bit images of logs >15cm diameters to determine dispersed wood residues in OR and compare to WRS methods. Across blocks, geospatial and WRS method wood volumes were correlated (R2=0.69), however volumes were 2.5 times larger for the geospatial vs WRS method. Methods for dispersed residues could not be properly compared as individual WRS plots were not georeferenced, only 12 plots were sampled in total, and low-resolution images poorly resolved logs. Thus, a new study in 2 cutblocks in the Northwest Bay (NWB) area acquired 2cm resolution RGB air-photography in 2014-15 using an Aeryon Sky Ranger UAV prior to and after burn pile construction. A total of 57 dispersed WRS plots and 24 WRS pile or accumulation plots were georeferenced and measured. Stero-pairs were used to generate point-clouds for pile bulk volumes. Images processed to 8-bit grey scale are being analyzed with a revised PCI method that better accounts for log overlaps. WRS methods depend on a good sample of plots and accurate determination of stratum (dispersed, roadside, piles, accumulations) areas. Analysis of NWB blocks shows WRS field methods for stratum area differ by 5-20% from that determined using orthophotos. Plot-level wood

  11. Scintigraphic determination of the spleen volume in case of partial splenectomy for a Minkowski-Chauffard disease. About 15 cases; Determination scintigraphique du volume splenique en cas de splenectomie partielle pour une maladie de Minkowski-Chauffard. A propos de 15 cas

    Energy Technology Data Exchange (ETDEWEB)

    Berehou, F.; Ben Rais, N. [CHU Ibn Sina, Rabat, Service de medecine nucleaire (Morocco); Calzada, M.; Keller, I.; Devaux, J. [Hopital Saint-Antoine, service de medecine nucleaire, 75 - Paris (France)

    2010-07-01

    Splenic scintigraphy with labeled and fragile red blood cells is a simple, reliable, noninvasive and low radiating method to assess at long-term residual splenic volume after partial splenectomy, it is the only treatment for a good quality of life with fewer infections, fewer transfusions and without use of lifelong vaccinations for a chronic illness. (N.C.)

  12. Stereological analysis of the mediodorsal thalamic nucleus in schizophrenia: volume, neuron number, and cell types

    DEFF Research Database (Denmark)

    Dorph-Petersen, Karl-Anton; Pierri, Joseph N; Sun, Zhuoxin

    2004-01-01

    The mediodorsal thalamic nucleus (MD) is the principal relay nucleus for the prefrontal cortex, a brain region thought to be dysfunctional in schizophrenia. Several, but not all, postmortem studies of the MD in schizophrenia have reported decreased volume and total neuronal number. However......, it is not clear whether the findings are specific for schizophrenia nor is it known which subtypes of thalamic neurons are affected. We studied the left MD in 11 subjects with schizophrenia, 9 control subjects, and 12 subjects with mood disorders. Based on morphological criteria, we divided the neurons into two...... subclasses, presumably corresponding to projection neurons and local circuit neurons. We estimated MD volume and the neuron number of each subclass using methods based on modern unbiased stereological principles. We also estimated the somal volumes of each subclass using a robust, but biased, approach...

  13. A new method to address unmet needs for extracting individual cell migration features from a large number of cells embedded in 3D volumes.

    Directory of Open Access Journals (Sweden)

    Ivan Adanja

    Full Text Available BACKGROUND: In vitro cell observation has been widely used by biologists and pharmacologists for screening molecule-induced effects on cancer cells. Computer-assisted time-lapse microscopy enables automated live cell imaging in vitro, enabling cell behavior characterization through image analysis, in particular regarding cell migration. In this context, 3D cell assays in transparent matrix gels have been developed to provide more realistic in vitro 3D environments for monitoring cell migration (fundamentally different from cell motility behavior observed in 2D, which is related to the spread of cancer and metastases. METHODOLOGY/PRINCIPAL FINDINGS: In this paper we propose an improved automated tracking method that is designed to robustly and individually follow a large number of unlabeled cells observed under phase-contrast microscopy in 3D gels. The method automatically detects and tracks individual cells across a sequence of acquired volumes, using a template matching filtering method that in turn allows for robust detection and mean-shift tracking. The robustness of the method results from detecting and managing the cases where two cell (mean-shift trackers converge to the same point. The resulting trajectories quantify cell migration through statistical analysis of 3D trajectory descriptors. We manually validated the method and observed efficient cell detection and a low tracking error rate (6%. We also applied the method in a real biological experiment where the pro-migratory effects of hyaluronic acid (HA were analyzed on brain cancer cells. Using collagen gels with increased HA proportions, we were able to evidence a dose-response effect on cell migration abilities. CONCLUSIONS/SIGNIFICANCE: The developed method enables biomedical researchers to automatically and robustly quantify the pro- or anti-migratory effects of different experimental conditions on unlabeled cell cultures in a 3D environment.

  14. Avaliação cronológica da variação no volume globular sanguíneo de bovinos leiteiros Cronological evaluation of variation in packed cell volume on dairy cattle

    Directory of Open Access Journals (Sweden)

    Wilmar Sachetin Marçal

    1995-01-01

    Full Text Available Os autores avaliaram a variação no volume globular sangüíneo de 321 bovinos da raça Holandês preta e branca, sadios e criados em granjas leiteiras no Estado de São Paulo. Todos os animais trabalhados na presente pesquisa eram sadios, não reagentes ao vírus da Leucose Bovina, livres de hemoparasitas, brucelose e tuberculose. O volume globular sangüíneo foi efetuado através do método do hematócrito com tubos capilares. Os resultados mostram haver influência da idade sobre o volume globular sangüíneo, com valores médios encontrados de 30,12 ± 2,72%.The packet cell volume was evaluated by the authors in 321 healthy female Holstein cattle raised at Campinas dairy region. São Paulo State. All the animais used in this assay were healthy and free of Leucosis, Tuberculosis, Brucellosis and blood parasites. The packed cell volume has been studied by method of hematocrit with capilary tubos. The results showed an influence of age on packed cell volume with average reference values of 30.12 ± 2.72%.

  15. Brownian Dynamics of a Suspension of Particles with Constrained Voronoi Cell Volumes

    KAUST Repository

    Singh, John P.

    2015-06-23

    © 2015 American Chemical Society. Solvent-free polymer-grafted nanoparticle fluids consist of inorganic core particles fluidized by polymers tethered to their surfaces. The attachment of the suspending fluid to the particle surface creates a strong penalty for local variations in the fluid volume surrounding the particles. As a model of such a suspension we perform Brownian dynamics of an equilibrium system consisting of hard spheres which experience a many-particle potential proportional to the variance of the Voronoi volumes surrounding each particle (E = α(Vi-V0)2). The coefficient of proportionality α can be varied such that pure hard sphere dynamics is recovered as α → 0, while an incompressible array of hairy particles is obtained as α →. As α is increased the distribution of Voronoi volumes becomes narrower, the mean coordination number of the particle increases and the variance in the number of nearest neighbors decreases. The nearest neighbor peaks in the pair distribution function are suppressed and shifted to larger radial separations as the constraint acts to maintain relatively uniform interstitial regions. The structure factor of the model suspension satisfies S(k=0) → 0 as α → in accordance with expectation for a single component (particle plus tethered fluid) incompressible system. The tracer diffusivity of the particles is reduced by the volume constraint and goes to zero at φ 0.52, indicating an earlier glass transition than has been observed in hard sphere suspensions. The total pressure of the suspension grows in proportion to (αkBT)1/2 as the strength of the volume-constraint potential grows. This stress arises primarily from the interparticle potential forces, while the hard-sphere collisional contribution to the stress is suppressed by the volume constraint.

  16. Brownian Dynamics of a Suspension of Particles with Constrained Voronoi Cell Volumes.

    Science.gov (United States)

    Singh, John P; Walsh, Stuart D C; Koch, Donald L

    2015-06-23

    Solvent-free polymer-grafted nanoparticle fluids consist of inorganic core particles fluidized by polymers tethered to their surfaces. The attachment of the suspending fluid to the particle surface creates a strong penalty for local variations in the fluid volume surrounding the particles. As a model of such a suspension we perform Brownian dynamics of an equilibrium system consisting of hard spheres which experience a many-particle potential proportional to the variance of the Voronoi volumes surrounding each particle (E = α(Vi-V0)(2)). The coefficient of proportionality α can be varied such that pure hard sphere dynamics is recovered as α → 0, while an incompressible array of hairy particles is obtained as α → ∞. As α is increased the distribution of Voronoi volumes becomes narrower, the mean coordination number of the particle increases and the variance in the number of nearest neighbors decreases. The nearest neighbor peaks in the pair distribution function are suppressed and shifted to larger radial separations as the constraint acts to maintain relatively uniform interstitial regions. The structure factor of the model suspension satisfies S(k=0) → 0 as α → ∞ in accordance with expectation for a single component (particle plus tethered fluid) incompressible system. The tracer diffusivity of the particles is reduced by the volume constraint and goes to zero at ϕ ∼ 0.52, indicating an earlier glass transition than has been observed in hard sphere suspensions. The total pressure of the suspension grows in proportion to (αkBT)(1/2) as the strength of the volume-constraint potential grows. This stress arises primarily from the interparticle potential forces, while the hard-sphere collisional contribution to the stress is suppressed by the volume constraint.

  17. Space-time adaptive ADER discontinuous Galerkin finite element schemes with a posteriori sub-cell finite volume limiting

    CERN Document Server

    Zanotti, Olindo; Dumbser, Michael; Hidalgo, Arturo

    2015-01-01

    In this paper we present a novel arbitrary high order accurate discontinuous Galerkin (DG) finite element method on space-time adaptive Cartesian meshes (AMR) for hyperbolic conservation laws in multiple space dimensions, using a high order \\aposteriori sub-cell ADER-WENO finite volume \\emph{limiter}. Notoriously, the original DG method produces strong oscillations in the presence of discontinuous solutions and several types of limiters have been introduced over the years to cope with this problem. Following the innovative idea recently proposed in \\cite{Dumbser2014}, the discrete solution within the troubled cells is \\textit{recomputed} by scattering the DG polynomial at the previous time step onto a suitable number of sub-cells along each direction. Relying on the robustness of classical finite volume WENO schemes, the sub-cell averages are recomputed and then gathered back into the DG polynomials over the main grid. In this paper this approach is implemented for the first time within a space-time adaptive ...

  18. Molecular determinants of treatment response in human germ cell tumors

    NARCIS (Netherlands)

    F. Mayer; J.A. Stoop (Hans); G.L. Scheffer (George); R. Scheper; J.W. Oosterhuis (Wolter); L.H.J. Looijenga (Leendert); C. Bokemeyer

    2003-01-01

    textabstractPURPOSE: Germ cell tumors (GCTs) are highly sensitive to cisplatin-based chemotherapy. This feature is unexplained, as is the intrinsic chemotherapy resistance of mature teratomas and the resistant phenotype of a minority of refractory GCTs. Various cellular pathways ma

  19. Time constant determination for electrical equivalent of biological cells

    Science.gov (United States)

    Dubey, Ashutosh Kumar; Dutta-Gupta, Shourya; Kumar, Ravi; Tewari, Abhishek; Basu, Bikramjit

    2009-04-01

    The electric field interactions with biological cells are of significant interest in various biophysical and biomedical applications. In order to study such important aspect, it is necessary to evaluate the time constant in order to estimate the response time of living cells in the electric field (E-field). In the present study, the time constant is evaluated by considering the hypothesis of electrical analog of spherical shaped cells and assuming realistic values for capacitance and resistivity properties of cell/nuclear membrane, cytoplasm, and nucleus. In addition, the resistance of cytoplasm and nucleoplasm was computed based on simple geometrical considerations. Importantly, the analysis on the basis of first principles shows that the average values of time constant would be around 2-3 μs, assuming the theoretical capacitance values and the analytically computed resistance values. The implication of our analytical solution has been discussed in reference to the cellular adaptation processes such as atrophy/hypertrophy as well as the variation in electrical transport properties of cellular membrane/cytoplasm/nuclear membrane/nucleoplasm.

  20. STRIPAK components determine mode of cancer cell migration and metastasis

    DEFF Research Database (Denmark)

    Madsen, Chris D; Hooper, Steven; Tozluoglu, Melda;

    2015-01-01

    and MST4 kinases, which promote the co-localization of the contractile actomyosin machinery with the Ezrin/Radixin/Moesin family proteins by phosphorylating the inhibitors of PPP1CB, PPP1R14A-D. Using computational modelling, in vitro cell migration assays and in vivo breast cancer metastasis assays we...

  1. Molecular determinants of treatment response in human germ cell tumors

    NARCIS (Netherlands)

    F. Mayer; J.A. Stoop (Hans); G.L. Scheffer (George); R. Scheper; J.W. Oosterhuis (Wolter); L.H.J. Looijenga (Leendert); C. Bokemeyer

    2003-01-01

    textabstractPURPOSE: Germ cell tumors (GCTs) are highly sensitive to cisplatin-based chemotherapy. This feature is unexplained, as is the intrinsic chemotherapy resistance of mature teratomas and the resistant phenotype of a minority of refractory GCTs. Various cellular pathways

  2. Quantitative Determination of Ceramide Molecular Species in Dendritic Cells

    Directory of Open Access Journals (Sweden)

    Samar Al Makdessi

    2016-09-01

    Full Text Available Background/Aims: The activation of acid sphingomyelinase by cellular stress or receptors or the de novo synthesis lead to the formation of ceramide (N-acylsphingosine, which in turn modifies the biophysical properties of cellular membrane and greatly amplifies the intensity of the initial signal. Ceramide, which acts by re-organizing a given signalosome rather than being a second messenger, has many functions in infection biology, cancer, cardiovascular syndromes, and immune regulation. Experimental studies on the infection of human cells with different bacterial agents demonstrated the activation of the acid sphingomyelinase/ceramide system. Moreover, the release of ceramide was found to be a requisite for the uptake of the pathogen. Considering the particular importance of the cellular role of ceramide, it was necessary to develop sensitive and accurate methods for its quantification. Methods: Here, we describe a method quantifying ceramide in dendritic cells and defining the different fatty acids (FA bound to sphingosine. The main steps of the method include extraction of total lipids, separation of the ceramide by thin-layer chromatography, derivatization of ceramide-fatty acids (Cer-FA, and quantitation of these acids in their methyl form by gas chromatography on polar capillary columns. The identification of FA was achieved by means of known standards and confirmed by mass spectrometry. Results: FA ranging between C10 and C24 could be detected and quantified. The concentration of the sum of Cer-FA amounted to 14.88 ± 8.98 nmol/106 cells (n=10. Oleic acid, which accounted for approximately half of Cer-FA (7.73 ± 6.52 nmol/106 cells was the predominant fatty acid followed by palmitic acid (3.47 ± 1.54 nmol/106 cells. Conclusion: This highly sensitive method allows the quantification of different molecular species of ceramides.

  3. Environmental determinants of severity in sickle cell disease.

    Science.gov (United States)

    Tewari, Sanjay; Brousse, Valentine; Piel, Frédéric B; Menzel, Stephan; Rees, David C

    2015-09-01

    Sickle cell disease causes acute and chronic illness, and median life expectancy is reduced by at least 30 years in all countries, with greater reductions in low-income countries. There is a wide spectrum of severity, with some patients having no symptoms and others suffering frequent, life-changing complications. Much of this variability is unexplained, despite increasingly sophisticated genetic studies. Environmental factors, including climate, air quality, socio-economics, exercise and infection, are likely to be important, as demonstrated by the stark differences in outcomes between patients in Africa and USA/Europe. The effects of weather vary with geography, although most studies show that exposure to cold or wind increases hospital attendance with acute pain. Most of the different air pollutants are closely intercorrelated, and increasing overall levels seem to correlate with increased hospital attendance, although higher concentrations of atmospheric carbon monoxide may offer some benefit for patients with sickle cell disease. Exercise causes some adverse physiological changes, although this may be off-set by improvements in cardiovascular health. Most sickle cell disease patients live in low-income countries and socioeconomic factors are undoubtedly important, but little studied beyond documenting that sickle cell disease is associated with decreases in some measures of social status. Infections cause many of the differences in outcomes seen across the world, but again these effects are relatively poorly understood. All the above factors are likely to account for much of the pathology and variability of sickle cell disease, and large prospective studies are needed to understand these effects better.

  4. Estimation of cell volume and biomass of penicillium chrysogenum using image analysis.

    Science.gov (United States)

    Packer, H L; Keshavarz-Moore, E; Lilly, M D; Thomas, C R

    1992-02-20

    A methodology for the estimation of biomass for the penicillin fermentation using image analysis is presented. Two regions of hyphae are defined to describe the growth of mycelia during fermentation: (1) the cytoplasmic region, and (2) the degenerated region including large vacuoles. The volume occupied by each of these regions in a fixed volume of sample is estimated from area measurements using image analysis. Areas are converted to volumes by treating the hyphae as solid cylinders with the hyphal diameter as the cylinder diameter. The volumes of the cytoplasmic and degenerated regions are converted into dry weight estimations using hyphal density values available from the literature. The image analysis technique is able to estimate biomass even in the presence of nondissolved solids of a concentration of up to 30 gL(-1). It is shown to estimate successfully concentrations of mycelia from 0.03 to 38 gL(-1). Although the technique has been developed for the penicillin fermentation, it should be applicable to other (nonpellected) fungal fermentations.

  5. GADD45β Determines Chemoresistance and Invasive Growth of Side Population Cells of Human Embryonic Carcinoma

    Directory of Open Access Journals (Sweden)

    Toshihiko Inowa

    2010-01-01

    Full Text Available Side population (SP cells are an enriched population of stem, and the existence of SP cells has been reported in human cancer cell lines. In this study, we performed an SP analysis using 11 human cancer cell lines and confirmed the presence of SP cells in an embryonic carcinoma cell line, NEC8. NEC8 SP cells showed characteristics of cancer stem cells, such as high growth rate, chemoresistance and high invasiveness. To further characterize the NEC8 SP cells, we used DNA microarrays. Among 38,500 genes, we identified 12 genes that were over-expressed in SP cells and 1 gene that was over-expressed in non-SP cells. Among these 13 genes, we focused on GADD45b. GADD45b was over-expressed in non-SP cells, but the inhibition of GADD45b had no effect on non-SP cells. Paradoxically, the inhibition of GADD45b significantly reduced the viability of NEC8 SP cells. The inhibition of ABCG2, which determines the SP phenotype, had no effect on the invasiveness of NEC8 SP cells, but the inhibition of GADD45b significantly reduced invasiveness. These results suggest that GADD45b, but not ABCG2, might determine the cancer stem cell-like phenotype, such as chemoresistance and the high invasiveness of NEC8 SP cells, and might be a good therapeutic target.

  6. Effect of different segmentation algorithms on metabolic tumor volume measured on 18F-FDG PET/CT of cervical primary squamous cell carcinoma

    Science.gov (United States)

    Xu, Weina; Yu, Shupeng; Ma, Ying; Liu, Changping

    2017-01-01

    Background and purpose It is known that fluorine-18 fluorodeoxyglucose PET/computed tomography (CT) segmentation algorithms have an impact on the metabolic tumor volume (MTV). This leads to some uncertainties in PET/CT guidance of tumor radiotherapy. The aim of this study was to investigate the effect of segmentation algorithms on the PET/CT-based MTV and their correlations with the gross tumor volumes (GTVs) of cervical primary squamous cell carcinoma. Materials and methods Fifty-five patients with International Federation of Gynecology and Obstetrics stage Ia∼IIb and histologically proven cervical squamous cell carcinoma were enrolled. A fluorine-18 fluorodeoxyglucose PET/CT scan was performed before definitive surgery. GTV was measured on surgical specimens. MTVs were estimated on PET/CT scans using different segmentation algorithms, including a fixed percentage of the maximum standardized uptake value (20∼60% SUVmax) threshold and iterative adaptive algorithm. We divided all patients into four different groups according to the SUVmax within target volume. The comparisons of absolute values and percentage differences between MTVs by segmentation and GTV were performed in different SUVmax subgroups. The optimal threshold percentage was determined from MTV20%∼MTV60%, and was correlated with SUVmax. The correlation of MTViterative adaptive with GTV was also investigated. Results MTV50% and MTV60% were similar to GTV in the SUVmax up to 5 (P>0.05). MTV30%∼MTV60% were similar to GTV (P>0.05) in the 50.05) in the 100.05) in the SUVmax of at least 15 group. MTViterative adaptive was similar to GTV in both total and different SUVmax groups (P>0.05). Significant differences were observed among the fixed percentage method and the optimal threshold percentage was inversely correlated with SUVmax. The iterative adaptive segmentation algorithm led to the highest accuracy (6.66±50.83%). A significantly positive correlation was also observed between MTViterative

  7. In vivo protein synthesis determinations in human immune cells

    OpenAIRE

    Januszkiewicz, Anna

    2005-01-01

    Intact immune responses are essential for defeating severe infections in individual patients. Insufficient function of the immune system contributes to a poor prognosis in these patients, in particular the ICU patients. Nevertheless, the immune system function is not easily monitored and evaluated. The ongoing metabolic activity of immune competent cells is reflected by their in vivo protein synthesis rate. The aim of this thesis was to apply in vivo protein synthesis measur...

  8. STRIPAK components determine mode of cancer cell migration and metastasis

    Science.gov (United States)

    Madsen, Chris D.; Hooper, Steven; Tozluoglu, Melda; Bruckbauer, Andreas; Fletcher, Georgina; Erler, Janine T.; Bates, Paul A.; Thompson, Barry; Sahai, Erik

    2017-01-01

    The contractile actomyosin cytoskeleton and its connection to the plasma membrane are critical for control of cell shape and migration. We identify three STRIPAK complex components, FAM40A, FAM40B, and STRN3, as regulators of the actomyosin cortex. We show that FAM40A negatively regulates the MST3 and MST4 kinases, which promote the co-localisation of the contractile actomyosin machinery with the Ezrin/Radixin/Moesin family proteins by phosphorylating the inhibitors of PPP1CB, PPP1R14A-D. Using computational modelling, in vitro cell migration assays and in vivo breast cancer metastasis assays we demonstrate that co-localisation of contractile activity and actin-plasma membrane linkage reduces cell speed on planar surfaces, but favours migration in confined environments similar to those observed in vivo. We further show that FAM40B mutations found in human tumours uncouple it from PP2A and enable it to drive a contractile phenotype, which may underlie its role in human cancer. PMID:25531779

  9. Determinants of action potential propagation in cerebellar Purkinje cell axons.

    Science.gov (United States)

    Monsivais, Pablo; Clark, Beverley A; Roth, Arnd; Häusser, Michael

    2005-01-12

    Axons have traditionally been viewed as highly faithful transmitters of action potentials. Recently, however, experimental evidence has accumulated to support the idea that under some circumstances axonal propagation may fail. Cerebellar Purkinje neurons fire highfrequency simple spikes, as well as bursts of spikes in response to climbing fiber activation (the "complex spike"). Here we have visualized the axon of individual Purkinje cells to directly investigate the relationship between somatic spikes and axonal spikes using simultaneous somatic whole-cell and cell-attached axonal patch-clamp recordings at 200-800 microm from the soma. We demonstrate that sodium action potentials propagate at frequencies up to approximately 260 Hz, higher than simple spike rates normally observed in vivo. Complex spikes, however, did not propagate reliably, with usually only the first and last spikes in the complex spike waveform being propagated. On average, only 1.7 +/- 0.2 spikes in the complex spike were propagated during resting firing, with propagation limited to interspike intervals above approximately 4 msec. Hyperpolarization improved propagation efficacy without affecting total axonal spike number, whereas strong depolarization could abolish propagation of the complex spike. These findings indicate that the complex spike waveform is not faithfully transmitted to downstream synapses and that propagation of the climbing fiber response may be modulated by background activity.

  10. In vitro isotopic determination of diffusion volumes by transferrin labelled with indium 111. study of the correlation with SARI 125; Determination isotopique in vitro de volumes de diffusion par la transferrine marquee a l'indium 111. Etude de la correlation avec la SARI 125

    Energy Technology Data Exchange (ETDEWEB)

    Porot, C.L. [CHU Jean-Minjoz, Service de medecine nucleaire, 25 - Besancon (France); Angoue, O.R. [CHU Jean-Minjoz, laboratoire de biophysique et statistiques, 25 - Besancon (France); Berthetc, L.O. [CHU Jean-Minjoz, 25 - Besancon (France); Ungureanu, C.O.; Boulahdour, H.A.

    2010-07-01

    Serum albumin labeled with iodine 125 (S.A.R.I. 125) is the reference tracer used in measuring isotopic plasma volume. It has been causing a suspension of manufacturing leading to a supply disruption and resulting in the search for an alternative to measure plasma volume under consideration for measuring blood volume. Plasma transferrin labeled with indium-111 (Tf-{sup 111}In) is a potentially useful marker. To this end, we assessed the level of activity to be administered to determine a volume of distribution. The study of the correlation between the volume of distribution values obtained with S.A.R.I. 125 and Tf-{sup 111}In was then performed. Tf is an autologous protein which the labelling is easy and stable. Tf-{sup 111}In is a valid alternative to the S.A.R.I. 125 for measuring the plasma volume. The activity required for this examination shall not exceed 100 micro curies. (N.C.)

  11. International review of cytology. Volume 109: A survey of cell biology

    Energy Technology Data Exchange (ETDEWEB)

    Bourne, G.; Jeon, K.W.; Friedlander, M.

    1987-01-01

    This book's contents are: Local Regulation of Testicular Function;Microtubules and DNA Replication;Differentiation of Spermatogenic Cells from Vertebrates in Vitro;The Developmental Program of Spermiogenesis in Drosophila: A Genetic Analysis;Cell Motility and Ionic Relations in Characean Cells as Revealed by Internal Perfusion and Other Cell Models;and The Culture of Oral Epithelium. Each chapter includes references.

  12. Liver volume measurement: reason of the difference between in vivo CT-volumetry and intraoperative ex vivo determination and how to cope it

    Directory of Open Access Journals (Sweden)

    Niehues SM

    2010-08-01

    Full Text Available Abstract Purpose Volumetric assessment of the liver regularly yields discrepant results between pre- and intraoperatively determined volumes. Nevertheless, the main factor responsible for this discrepancy remains still unclear. The aim of this study was to systematically determine the difference between in vivo CT-volumetry and ex vivo volumetry in a pig animal model. Material and Methods Eleven pigs were studied. Liver density assessment, CT-volumetry and water displacement volumetry was performed after surgical removal of the complete liver. Known possible errors of volume determination like resection or segmentation borders were eliminated in this model. Regression analysis was performed and differences between CT-volumetry and water displacement determined. Results Median liver density was 1.07 g/ml. Regression analysis showed a high correlation of r2 = 0.985 between CT-volumetry and water displacement. CTvolumetry was found to be 13% higher than water displacement volumetry (p Conclusion In this study the only relevant factor leading to the difference between in vivo CT-volumetry and ex vivo water displacement volumetry seems to be blood perfusion of the liver. The systematic difference of 13 percent has to be taken in account when dealing with those measures.

  13. Factors determining cell killing by chemotherapeutic agents In Vivo-II : Melphalan, chlorambucil and nitrogen mustard

    NARCIS (Netherlands)

    Putten, L.M. van; Lelieveld, P.

    1971-01-01

    The sensitivity of resting spleen colony-forming cells in normal mouse bone marrow and of rapidly-proliferating spleen colony-forming cells in repopulating mouse spleen to a number of bis-chloro-aethyl-amines was determined. The rapidly-growing cells were more sensitive to each of the agents than

  14. Determination of optimum electrolyte composition for molten carbonate fuel cells

    Energy Technology Data Exchange (ETDEWEB)

    Yuh, C.Y.; Pigeaud, A.

    1987-01-01

    The goal of this program is the development of a molten carbonate fuel cell (MCFC) anode which is more tolerant of sulfur contaminants in the fuel than the current state-of-the-art nickel-based anode structures. This program addresses two different but related aspects of the sulfur contamination problem. The primary aspect is concerned with the development of a sulfur tolerant electrocatalyst for the fuel oxidation reaction. A secondary issue is the development of a sulfur tolerant water-gas-shift reaction catalyst and an investigation of potential steam reforming catalysts which also have some sulfur tolerant capabilities. These two aspects are being addressed as two separate tasks.

  15. Determination of the Antibiotic Resistance Profile of Student Cell Phones

    Directory of Open Access Journals (Sweden)

    Lisa Ann Blankinship

    2012-08-01

    Full Text Available Sampling of common use items (e.g., student cell phones for bacterial presence, identification, and antibiotic resistance profiling helps students to recognize the need for routine cleaning of personal items and encourages thoughtful use of currently available medications. This multilab period project can be used to teach or reinforce several methods from general microbiology including aseptic technique, isolation streak, serial dilution, spread plating, Kirby Bauer testing, unknown identification, and media production. The data generated can be saved and added to each semester, thus providing a data set that reflects a local trend of antibiotic resistance.      

  16. Epidermal Patterning Genes Impose Non-cell Autonomous Cell Size Determination and have Additional Roles in Root Meristem Size Control

    Institute of Scientific and Technical Information of China (English)

    Christian L?fke; Kai Dünser; Jürgen Kleine-Vehn

    2013-01-01

    The regulation of cellular growth is of vital importance for embryonic and postembryonic patterning. Growth regulation in the epidermis has importance for organ growth rates in roots and shoots, proposing epidermal cells as an interesting model for cellular growth regulation. Here we assessed whether the root epidermis is a suitable model system to address cell size determination. In Arabidopsis thaliana L., root epidermal cells are regularly spaced in neighbouring tricho-(root hair) and atrichoblast (non-hair) cells, showing already distinct cell size regulation in the root meristem. We determined cell sizes in the root meristem and at the onset of cellular elongation, revealing that not only division rates but also cellular shape is distinct in tricho-and atrichoblasts. Intriguingly, epidermal-patterning mutants, failing to define differential vacuolization in neighbouring epidermal cell files, also display non-differential growth. Using these epidermal-patterning mutants, we show that polarized growth behaviour of epidermal tricho-and atrichoblast is interdependent, suggesting non-cell autonomous signals to integrate tissue expansion. Besides the interweaved cell-type-dependent growth mechanism, we reveal an additional role for epidermal patterning genes in root meristem size and organ growth regulation. We conclude that epidermal cells represent a suitable model system to study cell size determination and interdependent tissue growth.

  17. Design of flow chamber with electronic cell volume capability and light detection optics for multilaser flow cytometry.

    Science.gov (United States)

    Schuette, W H; Shackney, S E; Plowman, F A; Tipton, H W; Smith, C A; MacCollum, M A

    1984-11-01

    A multibeam optical detection system has been developed with a high optical efficiency, achieved through a reduction in the number of optical interfaces employed in the system. This reduction is made possible by a combination of employing simple lenses, gluing the objective lens directly upon the face of the flow cuvette and the extraction of only one fluorescence signal from each laser beam. A modified flow chamber is also described that includes fluidic resistance elements for the elimination of most of the electric shielding normally associated with electronic cell volume measurements.

  18. Experimental Evaluation of the Applicability of Capacitive and Optical Measurement Methods for the Determination of Liquid Hydrogen Volume Flow

    Directory of Open Access Journals (Sweden)

    Gert HOLLER

    2009-08-01

    Full Text Available This paper presents a capacitive and a vision-based method for measuring the velocity of cryogenic hydrogen flows. The capacitive sensing principle exploits the spatial frequency signature of perturbations moving through a multi-electrode structure. This setup increases the sensitivity to dielectric permittivity variations compared to a simple two-electrode structure while preserving the ability to detect small perturbations. The vision-based method relies on a high-speed camera system that monitors the liquid hydrogen flow through an optical window yielding the flow velocity by cross-correlating subsequent images of the flow. Although a comprehensive analysis of the obtainable measurement uncertainty was not performed yet, current measurement results show the applicability of both principles for the non-invasive measurement of the volume flow of cryogenic fuels inside conveyor pipes.

  19. The effect of strain path change on subgrain volume fraction determined from in situ X-ray measurements

    DEFF Research Database (Denmark)

    Wejdemann, Christian; Poulsen, Henning Friis; Lienert, U.

    2009-01-01

    The evolution of dislocation structures in individual bulk grains in copper during strain path changes is studied with a new in situ synchrotron technique which combines high angular resolution with fast three-dimensional reciprocal space mapping. Deformed copper contains regions with vanishing...... dislocation density called subgrains bounded by dislocation rich walls. With the new technique reciprocal space maps, consisting of sharp peaks arising from the subgrains superimposed on a cloud of lower intensity arising from the dislocation walls, are obtained, which allows properties such as subgrain...... volume fraction to be quantified. The studied strain path changes are tension-tension sequences. Polycrystalline copper sheets are pre-deformed in tension to 5% strain, and tensile samples are cut with varying angles between the first and second loading axis. The second tensile deformation up...

  20. Improved method to determine the molar volume and compositions of the NaCl-H2O-CO2 system inclusion

    Institute of Scientific and Technical Information of China (English)

    2007-01-01

    On the basis of Parry’s method (1986), an improved method was established to determine the molar volume (Vm) and compositions (X) of the NaCl-H2O-CO2 (NHC) system inclusion. To use this method, the determination of Vm-X only requires three microthermometric data of a NHC inclusion: partial homog-enization temperature (Th ,CO2), salinity (S) and total homogenization temperature (Th). Theoretically, four associated equations are needed containing four unknown parameters: X CO2, XNaCl, Vm and F (volume fraction of CO2 phase in total inclusion when occurring partial homogenization). When they are released, the Vm-X are determined. The former three equations, only correlated with Th ,CO2, S and F, have simplified expressions:XCO2=f1(Th,CO2,S,F),XNaCl=f2(Th,CO2,S,F),Vm=f3(Th,CO2,S,F). The last one is the thermodynamic relationship of X CO2, XNaCl, Vm and Th:f4(XCO2,XNaCl,Vm,Th)=0.Since the above four associated equations are complicated, it is necessary to adopt iterative technique to release them. The technique can be described by:(i) Freely input a F value (0≤F≤1),with Th ,CO2 and S, into the former three equations. As a result,X CO 2,XNaCl and the molar volume value recorded as Vm1 are derived. (ii) Input the X CO2 and XNaCl gotten in the step above into the last equation, and another molar volume value recorded as Vm2 is determined. (iii) If Vm1 is unequal to Vm2, the calculation will be restarted from “(i)”. The iteration is completed until Vm1 is equal to Vm2, which means that the four associated equations are released. Compared to Parry’s (1986) solution method, the improved method is more convenient to use, as well as more accurate to determine X CO 2. It is available for a NHC inlusion whose partial homogenization temperature is higher than clatherate melting temperature and there are no solid salt crystals in the inclusion at parital homogenization.

  1. Stochastic reconstruction and a scaling method to determine effective transport coefficients of a proton exchange membrane fuel cell catalyst layer

    Energy Technology Data Exchange (ETDEWEB)

    Barbosa, R. [Centro de Investigacion en Energia, UNAM, Privada Xochicalco S/N, 62580 Temixco (Mexico); Andaverde, J. [Centro de Investigacion en Ingenieria y Ciencias Aplicadas, UAEM, Av. Universidad 1001, Col. Chamilpa, 62210 Cuernavaca (Mexico); Escobar, B. [Instituto Tecnologico de Cancun, Av. Kabah 3, 77515 Cancun (Mexico); Cano, U. [Instituto de Investigaciones Electricas, Av. Reforma 113, col. Palmira, 62490 Cuernavaca (Mexico)

    2011-02-01

    This work uses a method for the stochastic reconstruction of catalyst layers (CLs) proposing a scaling method to determine effective transport properties in proton exchange membrane fuel cell (PEMFC). The algorithm that generates the numerical grid makes use of available information before and after manufacturing the CL. The structures so generated are characterized statistically by two-point correlation functions and by the resultant pore size distribution. As an example of this method, the continuity equation for charge transport is solved directly on the three-dimensional grid of finite control volumes (FCVs), to determine effective electrical and proton conductivities of different structures. The stochastic reconstruction and the electrical and proton conductivity of a 45 {mu}m side size cubic sample of a CL, represented by more than 3.3 x 10{sup 12} FVCs were realized in a much shorter time compared with non-scaling methods. Variables studied in an example of CL structure were: (i) volume fraction of dispersed electrolyte, (ii) total CL porosity and (iii) pore size distribution. Results for the conduction efficiency for this example are also presented. (author)

  2. Techniques for mapping the types, volumes, and distribution of clays in petroleum reservoirs and for determining their effects on oil production. Final report

    Energy Technology Data Exchange (ETDEWEB)

    Sharma, B.

    1993-05-01

    This report presents the results of correlation of log signatures with information on distribution of the types and volumes of clays in sandstone pore spaces determined from detailed CT-scan, XRD, SEM, and thin section analyses of core samples from three sandstone reservoirs. The log signatures are then analyzed to determine if suitable mathematical/statistical parameter(s) could be calculated from the logs to determine their effects on permeability and oil production. The variability measures obtained from power spectral analysis of permeability and wireline log data in clayey formations have been correlated with oil production from two oil fields. Compared with the conventional measures of permeability variations like the Dykstra-Parsons coefficients, the new measure appears to correlate better with oil production.

  3. Techniques for mapping the types, volumes, and distribution of clays in petroleum reservoirs and for determining their effects on oil production

    Energy Technology Data Exchange (ETDEWEB)

    Sharma, B.

    1993-05-01

    This report presents the results of correlation of log signatures with information on distribution of the types and volumes of clays in sandstone pore spaces determined from detailed CT-scan, XRD, SEM, and thin section analyses of core samples from three sandstone reservoirs. The log signatures are then analyzed to determine if suitable mathematical/statistical parameter(s) could be calculated from the logs to determine their effects on permeability and oil production. The variability measures obtained from power spectral analysis of permeability and wireline log data in clayey formations have been correlated with oil production from two oil fields. Compared with the conventional measures of permeability variations like the Dykstra-Parsons coefficients, the new measure appears to correlate better with oil production.

  4. Investigation of Adaptive-threshold Approaches for Determining Area-Time Integrals from Satellite Infrared Data to Estimate Convective Rain Volumes

    Science.gov (United States)

    Smith, Paul L.; VonderHaar, Thomas H.

    1996-01-01

    The principal goal of this project is to establish relationships that would allow application of area-time integral (ATI) calculations based upon satellite data to estimate rainfall volumes. The research is being carried out as a collaborative effort between the two participating organizations, with the satellite data analysis to determine values for the ATIs being done primarily by the STC-METSAT scientists and the associated radar data analysis to determine the 'ground-truth' rainfall estimates being done primarily at the South Dakota School of Mines and Technology (SDSM&T). Synthesis of the two separate kinds of data and investigation of the resulting rainfall-versus-ATI relationships is then carried out jointly. The research has been pursued using two different approaches, which for convenience can be designated as the 'fixed-threshold approach' and the 'adaptive-threshold approach'. In the former, an attempt is made to determine a single temperature threshold in the satellite infrared data that would yield ATI values for identifiable cloud clusters which are closely related to the corresponding rainfall amounts as determined by radar. Work on the second, or 'adaptive-threshold', approach for determining the satellite ATI values has explored two avenues: (1) attempt involved choosing IR thresholds to match the satellite ATI values with ones separately calculated from the radar data on a case basis; and (2) an attempt involved a striaghtforward screening analysis to determine the (fixed) offset that would lead to the strongest correlation and lowest standard error of estimate in the relationship between the satellite ATI values and the corresponding rainfall volumes.

  5. A method for rapid in situ extraction and laboratory determination of Th, Pb, and Ra isotopes from large volumes of seawater

    Science.gov (United States)

    Baskaran, M.; Murphy, David J.; Santschi, Peter H.; Orr, James C.; Schink, David R.

    1993-04-01

    An in situ pump and six channel extraction system, operating typically at 351 min -1, can not only filter seawater samples but also extract dissolved Th, Pb and Ra isotopes from large volumes of seawater at six different depths on a single lowering. By gamma counting the ash residues of three cartridges (one particle extractor and two nuclide extractors), we can determine 234Th and 210Pb concentrations in the particulate and dissolved phase. Extraction efficiencies for 234Th from the dissolved phase average 93 ± 5% and for 210Pb average ≥90%. At the same time we can determine radium isotopic ratios precisely, but, at present radium concentrations can only be established by the determination of 226Ra in a 30-1 cohort sample. This system can be deployed easily from ships having an A-frame. It eliminates the problems of transporting and processing large volumes of samples and involves relatively little manual labor and analytical time. We have tested and used this device repeatedly in the Gulf of Mexico. Our preliminary results agree well with values of 234Th and 210Pb concentrations and 228Ra/ 226Ra activity ratios reported in the literature for the Gulf of Mexico.

  6. PKB and megalin determine the survival or death of renal proximal tubule cells

    OpenAIRE

    Caruso-Neves, Celso; Pinheiro, Ana Acacia S.; Cai, Hui; Souza-Menezes, Jackson; Guggino, William B.

    2006-01-01

    Renal proximal tubule cells have a remarkable ability to reabsorb large quantities of albumin through megalin-mediated endocytosis. This is an essential process for overall body homeostasis. Overstressing this endocytic system with a prolonged excess of albumin is injurious to proximal tubule cells. How these cells function and protect themselves from injury is unknown. Here, we show that megalin is the sensor that determines whether cells will be protected or injured by albumin. Megalin, thr...

  7. Waves in cell monolayer without proliferation: density determines cell velocity and wave celerity

    CERN Document Server

    Tlili, S; Li, B; Cardoso, O; Ladoux, B; Delanoë-Ayari, H; Graner, F

    2016-01-01

    Collective cell migration contributes to morphogenesis, wound healing or tumor metastasis. Culturing epithelial monolayers on a substrate is an in vitro configuration suitable to quantitatively characterize such tissue migration by measuring cell velocity, density and cell-substrate interaction force. Inhibiting cell division, we limit cell density increase and favor steady cell migration, while by using long narrow strips we stabilise the migrating front shape, so that we observe migration over a day or more. In the monolayer bulk, the cell velocity is a function of the cell density, namely it increases as a linear function of the cell radius. At least ten periods of propagating velocity waves are detected with a high signal-to-noise ratio, enabling for their quantitative spatio-temporal analysis. Cell density displays waves, in phase opposition with the velocity, as predicted by mass conservation; similarly, cell-substrate force appear to display small amplitude waves, in phase quadrature with respect to ve...

  8. Parallel Changes in Intracellular Water Volume and pH Induced by NH3/NH4+ Exposure in Single Neuroblastoma Cells

    Directory of Open Access Journals (Sweden)

    Víctor M. Blanco

    2013-12-01

    Full Text Available Background: Increased blood levels of ammonia (NH3 and ammonium (NH4+, i.e. hyperammonemia, leads to cellular brain edema in humans with acute liver failure. The pathophysiology of this edema is poorly understood. This is partly due to incomplete understanding of the osmotic effects of the pair NH3/NH4+ at the cellular and molecular levels. Cell exposure to solutions containing NH3/NH4+ elicits changes in intracellular pH (pHi, which can in turn affect cell water volume (CWV by activating transport mechanisms that produce net gain or loss of solutes and water. The occurrence of CWV changes caused by NH3/NH4+ has long been suspected, but the mechanisms, magnitude and kinetics of these changes remain unknown. Methods: Using fluorescence imaging microscopy we measured, in real time, parallel changes in pHi and CWV caused by brief exposure to NH3/NH4+ of single cells (N1E-115 neuroblastoma or NG-108 neuroblastoma X glioma loaded with the fluorescent indicator BCECF. Changes in CWV were measured by exciting BCECF at its intracellular isosbestic wavelength (∼438 nm, and pHi was measured ratiometrically. Results: Brief exposure to isosmotic solutions (i.e. having the same osmolality as that of control solutions containing NH4Cl (0.5- 30 mM resulted in a rapid, dose-dependent swelling, followed by isosmotic regulatory volume decrease (iRVD. NH4Cl solutions in which either extracellular [NH3] or [NH4+] was kept constant while the other was changed by varying the pH of the solution, demonstrated that [NH3]o rather than [NH4+]o is the main determinant of the NH4Cl-induced swelling. The iRVD response was sensitive to the anion channel blocker NPPB, and partly dependent on external Ca2+. Upon removal of NH4Cl, cells shrank and displayed isosmotic regulatory volume increase (iRVI. Regulatory volume responses could not be activated by comparable CWV changes produced by anisosmotic solutions, suggesting that membrane stretch or contraction by themselves are

  9. Extracellular Regulation of the Mitotic Spindle and Fate Determinants Driving Asymmetric Cell Division.

    Science.gov (United States)

    Smith, Prestina; Azzam, Mark; Hinck, Lindsay

    2017-01-01

    Stem cells use mode of cell division, symmetric (SCD) versus asymmetric (ACD), to balance expansion with self-renewal and the generation of daughter cells with different cell fates. Studies in model organisms have identified intrinsic mechanisms that govern this process, which involves partitioning molecular components between daughter cells, frequently through the regulation of the mitotic spindle. Research performed in vertebrate tissues is revealing both conservation of these intrinsic mechanisms and crucial roles for extrinsic cues in regulating the frequency of these divisions. Morphogens and positional cues, including planar cell polarity proteins and guidance molecules, regulate key signaling pathways required to organize cell/ECM contacts and spindle pole dynamics. Noncanonical WNT7A/VANGL2 signaling governs asymmetric cell division and the acquisition of cell fates through spindle pole orientation in satellite stem cells of regenerating muscle fibers. During cortical neurogenesis, the same pathway regulates glial cell fate determination by regulating spindle size, independent of its orientation. Sonic hedgehog (SHH) stimulates the symmetric expansion of cortical stem and cerebellar progenitor cells and contributes to cell fate acquisition in collaboration with Notch and Wnt signaling pathways. SLIT2 also contributes to stem cell homeostasis by restricting ACD frequency through the regulation of spindle orientation. The capacity to influence stem cells makes these secreted factors excellent targets for therapeutic strategies designed to enhance cell populations in degenerative disease or restrict cell proliferation in different types of cancers.

  10. Experimental evidence for negative turgor pressure in small leaf cells of Robinia pseudoacacia L versus large cells of Metasequoia glyptostroboides Hu et W.C. Cheng. 2. Höfler diagrams below the volume of zero turgor and the theoretical implication for pressure-volume curves of living cells.

    Science.gov (United States)

    Yang, Dongmei; Li, Junhui; Ding, Yiting; Tyree, Melvin T

    2017-03-01

    The physiological advantages of negative turgor pressure, Pt , in leaf cells are water saving and homeostasis of reactants. This paper advances methods for detecting the occurrence of negative Pt in leaves. Biomechanical models of pressure-volume (PV) curves predict that negative Pt does not change the linearity of PV curve plots of inverse balance pressure, PB , versus relative water loss, but it does predict changes in either the y-intercept or the x-intercept of the plots depending on where cell collapse occurs in the PB domain because of negative Pt . PV curve analysis of Robinia leaves revealed a shift in the x-intercept (x-axis is relative water loss) of PV curves, caused by negative Pt of palisade cells. The low x-intercept of the PV curve was explained by the non-collapse of palisade cells in Robinia in the PB domain. Non-collapse means that Pt smoothly falls from positive to negative values with decreasing cell volume without a dramatic change in slope. The magnitude of negative turgor in non-collapsing living cells was as low as -1.3 MPa and the relative volume of the non-collapsing cell equaled 58% of the total leaf cell volume. This study adds to the growing evidence for negative Pt . © 2016 John Wiley & Sons Ltd.

  11. An optimization protocol for Swiss 3T3 feeder cell growth-arrest by Mitomycin C dose-to-volume derivation strategy.

    Science.gov (United States)

    Chugh, Rishi Man; Chaturvedi, Madhusudan; Yerneni, Lakshmana Kumar

    2017-04-01

    Feeder cell functionality following growth-arrest with the cost-effective Mitomycin C vis-à-vis irradiation is controversial due to several methodological variables reported. Earlier, we demonstrated variability in growth arrested Swiss 3T3 feeder cell life-span following titration of feeder cell densities with Mitomycin C concentrations which led to the derivation of doses per cell. Alternatively, to counter the unexpected feeder regrowth at high exposure cell density, we proposed titration of a fixed density with arithmetically derived volumes of Mitomycin C solution that corresponded to permutations of specific concentrations and doses per cell. We now describe an experimental procedure of inducing differential feeder cell growth-arrest by titrating with such volumes and validating the best feeder batch through target cell growth assessment. A safe cell density of Swiss 3T3 tested for the exclusion of Mitomycin C resistant variants was titrated with a range of volumes of a Mitomycin C solution. The differentially growth-arrested feeder batches generated were tested for short-term and long-term viability and human epidermal keratinocyte growth supporting ability. The feeder cell extinction rate was directly proportional to the volume of Mitomycin C solution within a given concentration per se. The keratinocyte colony forming efficiency and the overall growth in mass cultures were maximal with a median extinction rate produced by an intermediate volume, while the faster and slower extinction rates by high and low volumes, respectively, were suboptimal. The described method could counter the inadequacies of growth-arrest with Mitomycin C.

  12. Determination of ice thickness, subice topography and ice volume at Glacier No. 1 in Tien Shan, China by ground penetrating radar

    Institute of Scientific and Technical Information of China (English)

    孙波; 张鹏; 焦克勤; 邓新生; 温家洪

    2003-01-01

    We describe a radio-echo sounding (RES) survey for the determination of ice thickness, subglacial topography and ice volume of Glacier No. 1, in Tien Shah,China, using ground-penetrating radar (GPR). Radar data were collected with 100-MHz antennas that were spaced at 4 m with a step size of 8 m. The images produced from radar survey clearly show the continuity of bedrock echoes and the undulation features of the bedrock surface. Radar results show that the maximum ice thickness of Glacier No. 1 is 133 m, the thickness of the east branch of Glacier No. 1 averages at 58.77 m while that of the west branch of Glacier No. 1 averages at 44.84 m. Calculation on ice volume indicates that the ice volume of the east branch of Glacier No. 1 is 51.87 × 106 m3 and that of the west branch of Glacier No. 1 is 20.21 × 106 m3. The amplitude of the undulation of the bedrock surface topography revealed by radar profiles is larger than that of the glacier surface topography, indicating that the surface relief does not directly depend on that of the bedrock undulation in Glacier No. 1, in Tien Shan.

  13. [Determination of trace and ultra-trace level bromate in water by large volume sample injection with enrichment column for on-line preconcentration coupled with ion chromatography].

    Science.gov (United States)

    Liu, Jing; He, Qingqing; Yang, Lili; Hu, Enyu; Wang, Meifei

    2015-10-01

    A method for the determination of trace and ultra-trace level bromate in water by ion chromatography with large volume sample injection for on-line preconcentration was established. A high capacity Dionex IonPac AG23 guard column was simply used as the enrichment column instead of the loop for the preconcentration of bromate. High purity KOH solution used as eluent for gradient elution was on-line produced by an eluent generator automatically. The results showed that a good linear relationship of bromate was exhibited in the range of 0.05-51.2 μg/L (r ≥ 0.999 5), and the method detection limit was 0.01 μg/L. Compared with conventional sample injection, the injection volume was up to 5 mL, and the enrichment factor of this method was about 240 times. This method was successfully applied for several real samples of pure water which were purchased in the supermarket, and the recoveries of bromate were between 90%-100% with the RSDs (n = 6) of 2.1%-6.4% at two spiked levels. This method without pretreatment is simple, and of high accuracy and precision. The preconcentration can be achieved by large volume sample injection. It is suitable for the analysis of trace and ultra-trace level bromate.

  14. Serum thyroxine and age - rather than thyroid volume and serum TSH - are determinants of the thyroid radioiodine uptake in patients with nodular goiter

    DEFF Research Database (Denmark)

    Bonnema, S J; Fast, S; Nielsen, V E;

    2011-01-01

    patients (146 females; age range: 22-87 yrs.) with nodular goiter (median 64 ml, range: 20-464 ml) selected for 131I therapy. Serum TSH was sub-normal in 42.4%. None were treated with anti-thyroid drugs. The thyroid RAIU was determined at 24h and 96h. The goiter volume was measured by ultrasound (n=127......Background: 131I therapy is widely used for treatment of non-toxic goiters. A limitation for this treatment is a low thyroid radioiodine uptake (RAIU), often encountered in these patients. Aim: To estimate the impact of various factors on the thyroid RAIU. Methods: We examined prospectively 170...

  15. [Cell-ELA-based determination of binding affinity of DNA aptamer against U87-EGFRvIII cell].

    Science.gov (United States)

    Tan, Yan; Liang, Huiyu; Wu, Xidong; Gao, Yubo; Zhang, Xingmei

    2013-05-01

    A15, a DNA aptamer with binding specificity for U87 glioma cells stably overexpressing the epidermal growth factor receptor variant III (U87-EGFRvIII), was generated by cell systematic evolution of ligands by exponential enrichment (cell-SELEX) using a random nucleotide library. Subsequently, we established a cell enzyme-linked assay (cell-ELA) to detect the affinity of A15 compared to an EGFR antibody. We used A15 as a detection probe and cultured U87-EGFRvIII cells as targets. Our data indicate that the equilibrium dissociation constants (K(d)) for A15 were below 100 nmol/L and had similar affinity compared to an EGFR antibody for U87-EGFRvIII. We demonstrated that the cell-ELA was a useful method to determine the equilibrium dissociation constants (K(d)) of aptamers generated by cell-SELEX.

  16. CD8+ T Lymphocyte Self-Renewal during Effector Cell Determination

    Directory of Open Access Journals (Sweden)

    Wen-Hsuan W. Lin

    2016-11-01

    Full Text Available Selected CD8+ T cells must divide, produce differentiated effector cells, and self-renew, often repeatedly. We now show that silencing expression of the transcription factor TCF1 marks loss of self-renewal by determined effector cells and that this requires cell division. In acute infections, the first three CD8+ T cell divisions produce daughter cells with unequal proliferative signaling but uniform maintenance of TCF1 expression. The more quiescent initial daughter cells resemble canonical central memory cells. The more proliferative, effector-prone cells from initial divisions can subsequently undergo division-dependent production of a TCF1-negative effector daughter cell along with a self-renewing TCF1-positive daughter cell, the latter also contributing to the memory cell pool upon resolution of infection. Self-renewal in the face of effector cell determination may promote clonal amplification and memory cell formation in acute infections, sustain effector regeneration during persistent subclinical infections, and be rate limiting, but remediable, in chronic active infections and cancer.

  17. A numerical study on the elastic modulus of volume and area dilation for a deformable cell in a microchannel.

    Science.gov (United States)

    Moon, Ji Young; Tanner, Roger I; Lee, Joon Sang

    2016-07-01

    A red blood cell (RBC) in a microfluidic channel is highly interesting for scientists in various fields of research on biological systems. This system has been studied extensively by empirical, analytical, and numerical methods. Nonetheless, research of predicting the behavior of an RBC in a microchannel is still an interesting area. The complications arise from deformation of an RBC and interactions among the surrounding fluid, wall, and RBCs. In this study, a pressure-driven RBC in a microchannel was simulated with a three-dimensional lattice Boltzmann method of an immersed boundary. First, the effect of boundary thickness on the interaction between the wall and cell was analyzed by measuring the time of passage through the narrow channel. Second, the effect of volume conservation stiffness was studied. Finally, the effect of global area stiffness was analyzed.

  18. Preparation of cell blocks for lung cancer diagnosis and prediction: protocol and experience of a high-volume center.

    Science.gov (United States)

    Kossakowski, Claudia A; Morresi-Hauf, Alicia; Schnabel, Philipp A; Eberhardt, Ralf; Herth, Felix J F; Warth, Arne

    2014-01-01

    Minimally invasive diagnostic techniques are increasingly being used to obtain specimens for pathological diagnosis and prediction. Referring to lung cancer, both endobronchial and endoesophageal ultrasound are used worldwide as diagnostic routine methods. Consequently, an increasing number of pathological samples are cytological and fewer are histological. On the other hand, the requirements for specific and sensitive tumor subtyping complemented by predictive analyses are steadily increasing and are an essential basis for evidence-based treatment decisions. In this article we focus on the cell block method as a helpful tool for diagnostic and predictive analyses in lung cancer and point out its advantages and disadvantages in comparison to conventional cytological and biopsy specimens. Furthermore, we retrospectively analyze the diagnostic results of the cell block method in a high-volume center over 5 years. The main advantages of cell blocks are the availability of established and validated protocols, archiving and the opportunity to have serial sections from the same specimens to provide or repeat molecular analyses. Actually, in case of tumor progression, even additional biomarkers can be tested using the original cell block when re-biopsies are not feasible. The cell block method should be considered as a reliable, complimentary approach to conventional cytological or biopsy procedures, which is helpful to fulfill the increasing requirements of high-quality diagnostics and prediction.

  19. Establishment of HEK293 cell line expressing GFPAQP1 to determine water osmotic permeability

    Institute of Scientific and Technical Information of China (English)

    Jun-weiGAO; He-mingYU; Qian-liuSONG; Shu-xinLI; Xue-junLI

    2004-01-01

    AIM: To develop an osmotic water permeability assay.METHODS: We subcloned the rat AQP1 cDNA into pEGFPC3 vector. HEK293 cells were transfected with pEGFP-C3/AQP 1 or pEGFP-C3 and selected by G418. The expression of AQP1was detected by RT-PCR and Western blot. Confocal laser fluorescence microscopy was used to record the change of fluorescent density corresponding to the volume change of the cells

  20. Cell fate after mitotic arrest in different tumor cells is determined by the balance between slippage and apoptotic threshold

    Energy Technology Data Exchange (ETDEWEB)

    Galán-Malo, Patricia; Vela, Laura; Gonzalo, Oscar; Calvo-Sanjuán, Rubén; Gracia-Fleta, Lucía; Naval, Javier; Marzo, Isabel, E-mail: imarzo@unizar.es

    2012-02-01

    Microtubule poisons and other anti-mitotic drugs induce tumor death but the molecular events linking mitotic arrest to cell death are still not fully understood. We have analyzed cell fate after mitotic arrest produced by the microtubule-destabilizing drug vincristine in a panel of human tumor cell lines showing different response to vincristine. In Jurkat, RPMI 8226 and HeLa cells, apoptosis was triggered shortly after vincristine-induced mitotic arrest. However, A549 cells, which express a great amount of Bcl-x{sub L} and undetectable amounts of Bak, underwent mitotic slippage prior to cell death. However, when Bcl-x{sub L} gene was silenced in A549 cells, vincristine induced apoptosis during mitotic arrest. Another different behavior was found in MiaPaca2 cells, where vincristine caused death by mitotic catastrophe that switched to apoptosis when cyclin B1 degradation was prevented by proteasome inhibition. Overexpression of Bcl-x{sub L} or silencing Bax and Bak expression delayed the onset of apoptosis in Jurkat and RPMI 8226 cells, enabling mitotic slippage and endoreduplication. In HeLa cells, overexpression of Bcl-x{sub L} switched cell death from apoptosis to mitotic catastrophe. Mcl-1 offered limited protection to vincristine-induced cell death and Mcl-1 degradation was not essential for vincristine-induced death. All these results, taken together, indicate that the Bcl-x{sub L}/Bak ratio and the ability to degrade cyclin B1 determine cell fate after mitotic arrest in the different tumor cell types. Highlights: ► Vincristine induces cell death by apoptosis or mitotic catastrophe. ► Apoptosis-proficient cells die by apoptosis during mitosis upon vincristine treatment. ► p53wt apoptosis-deficient cells undergo apoptosis from a G1-like tetraploid state. ► p53mt apoptosis-deficient cells can survive and divide giving rise to 8N cells.

  1. Antioxidants and NOS inhibitors selectively targets manganese-induced cell volume via Na-K-Cl cotransporter-1 in astrocytes.

    Science.gov (United States)

    Alahmari, Khalid A; Prabhakaran, Harini; Prabhakaran, Krishnan; Chandramoorthy, Harish C; Ramugounder, Ramakrishnan

    2015-06-12

    Manganese has shown to be involved in astrocyte swelling. Several factors such as transporters, exchangers and ion channels are attributed to astrocyte swelling as a result in the deregulation of cell volume. Products of oxidation and nitration have been implied to be involved in the pathophysiology of swelling; however, the direct link and mechanism of manganese induced astrocyte swelling has not been fully elucidated. In the current study, we used rat primary astrocyte cultures to investigate the activation of Na-K-Cl cotransporter-1 (NKCC1) a downstream mechanism for free radical induced astrocyte swelling as a result of manganese toxicity. Our results showed manganese, oxidants and NO donors as potent inducer of oxidation and nitration of NKCC1. Our results further confirmed that manganese (50 μM) increased the total protein, phosphorylation and activity of NKCC1 as well as cell volume (p manganese or oxidants and NO induced activation, oxidation/nitration of NKCC1 play an important role in the astrocyte swelling. Copyright © 2015 Elsevier B.V. All rights reserved.

  2. Pressure-volume curves: revisiting the impact of negative turgor during cell collapse by literature review and simulations of cell micromechanics.

    Science.gov (United States)

    Ding, Yiting; Zhang, Yanxiang; Zheng, Quan-Shui; Tyree, Melvin T

    2014-07-01

    The Scholander-Hammel pressure chamber has been used in thousands of papers to measure osmotic pressure, πc , turgor pressure, Pt , and bulk modulus of elasticity, ε, of leaf cells by pressure-volume (PV) curve analysis. PV analysis has been questioned in the past. In this paper we use micromechanical analysis of leaf cells to examine the impact on PV curve analysis of negative turgor in living cells (Pt ). Models predict negative Pt (-0.1 to -1.8 MPa) depending on leaf cell size and shape in agreement with experimental values reported by J. J. Oertli. Modeled PV curves have linear regions even when Pt is quite negative, contrary to the arguments of M.T. Tyree. Negative Pt is totally missed by PV curve analysis and results in large errors in derived πc and Pt but smaller errors in ε. A survey of leaf cell sizes vs habitat (arid, temperate, and rainforest), suggests that the majority of published PV curves result in errors of 0.1-1.8 MPa in derived πc and Pt , whereby the error increases with decreasing cell size. We propose that small cell size in leaves is an ecological adaptation that permits plants to endure negative values of water potential with relatively little water loss. © 2014 The Authors. New Phytologist © 2014 New Phytologist Trust.

  3. Use of combined maximum and minimum intensity projections to determine internal target volume in 4-dimensional CT scans for hepatic malignancies

    Directory of Open Access Journals (Sweden)

    Liu Jin

    2012-01-01

    Full Text Available Abstract Background To evaluate the accuracy of the combined maximum and minimum intensity projection-based internal target volume (ITV delineation in 4-dimensional (4D CT scans for liver malignancies. Methods 4D CT with synchronized IV contrast data were acquired from 15 liver cancer patients (4 hepatocellular carcinomas; 11 hepatic metastases. We used five approaches to determine ITVs: (1. ITVAllPhases: contouring gross tumor volume (GTV on each of 10 respiratory phases of 4D CT data set and combining these GTVs; (2. ITV2Phase: contouring GTV on CT of the peak inhale phase (0% phase and the peak exhale phase (50% and then combining the two; (3. ITVMIP: contouring GTV on MIP with modifications based on physician's visual verification of contours in each respiratory phase; (4. ITVMinIP: contouring GTV on MinIP with modification by physician; (5. ITV2M: combining ITVMIP and ITVMinIP. ITVAllPhases was taken as the reference ITV, and the metrics used for comparison were: matching index (MI, under- and over-estimated volume (Vunder and Vover. Results 4D CT images were successfully acquired from 15 patients and tumor margins were clearly discernable in all patients. There were 9 cases of low density and 6, mixed on CT images. After comparisons of metrics, the tool of ITV2M was the most appropriate to contour ITV for liver malignancies with the highest MI of 0.93 ± 0.04 and the lowest proportion of Vunder (0.07 ± 0.04. Moreover, tumor volume, target motion three-dimensionally and ratio of tumor vertical diameter over tumor motion magnitude in cranio-caudal direction did not significantly influence the values of MI and proportion of Vunder. Conclusion The tool of ITV2M is recommended as a reliable method for generating ITVs from 4D CT data sets in liver cancer.

  4. Rhodopsin gene expression determines rod outer segment size and rod cell resistance to a dominant-negative neurodegeneration mutant.

    Directory of Open Access Journals (Sweden)

    Brandee A Price

    Full Text Available Two outstanding unknowns in the biology of photoreceptors are the molecular determinants of cell size, which is remarkably uniform among mammalian species, and the mechanisms of rod cell death associated with inherited neurodegenerative blinding diseases such as retinitis pigmentosa. We have addressed both questions by performing an in vivo titration with rhodopsin gene copies in genetically engineered mice that express only normal rhodopsin or an autosomal dominant allele, encoding rhodopsin with a disease-causing P23H substitution. The results reveal that the volume of the rod outer segment is proportional to rhodopsin gene expression; that P23H-rhodopsin, the most common rhodopsin gene disease allele, causes cell death via a dominant-negative mechanism; and that long term survival of rod cells carrying P23H-rhodopsin can be achieved by increasing the levels of wild type rhodopsin. These results point to promising directions in gene therapy for autosomal dominant neurodegenerative diseases caused by dominant-negative mutations.

  5. Gluten-free diet increases beta-cell volume and improves glucose tolerance in an animal model of type 2 diabetes.

    Science.gov (United States)

    Haupt-Jorgensen, Martin; Buschard, Karsten; Hansen, Axel K; Josefsen, Knud; Antvorskov, Julie Christine

    2016-10-01

    Gluten-free (GF) diet alleviates type 1 diabetes in animal models and possibly in humans. We recently showed that fatty acid-induced insulin secretion is enhanced by enzymatically digested gluten (gliadin) stimulation in INS-1E insulinoma cells. We therefore hypothesized that GF diet would induce beta-cell rest and ameliorate type 2 diabetes. C57BL/6JBomTac (B6) mice were fed a high-fat (HF), gluten-free high-fat (GF-HF), standard (STD) or gluten-free (GF) diet for 42 weeks. Short-term (6-24 weeks) GF-HF versus HF feeding impaired glucose tolerance and increased fasting glucose. Long-term (36-42 weeks) GF-HF versus HF feeding improved glucose tolerance and decreased fasting leptin. Mice fed a GF-HF versus HF diet for 42 weeks showed higher volumes of beta cells, islets and pancreas. The beta-cell volume correlated with the islet- and pancreas volume as well as body weight. GF-HF versus HF diet did not influence toll-like receptor 4 (Tlr4), interleukin 1 (IL-1), interleukin 6 (IL-6) or tumour necrosis factor-alpha (TNF-alpha) mRNA expression in intestine. STD versus GF feeding did not affect any parameter studied. Long-term feeding with GF-HF versus HF increases beta-cell volume and improves glucose tolerance in B6 mice. The mechanism may include beta-cell rest, but is unlikely to include TLR4 and proinflammatory cytokines in the intestine. Beta-cell volume correlates with pancreas volume and body weight, indicating that insulin secretion capacity controls pancreas volume. Thus, long-term GF diets may be beneficial for obese type 2 diabetes patients and trials should be performed. Copyright © 2016 John Wiley & Sons, Ltd. Copyright © 2016 John Wiley & Sons, Ltd.

  6. Determinants of Proteolysis and Cell-Binding for the Shigella flexneri Cytotoxin, SigA.

    Science.gov (United States)

    Chua, Eng Guan; Al-Hasani, Keith; Scanlon, Martin; Adler, Ben; Sakellaris, Harry

    2015-11-01

    Shigella flexneri secretes an enterotoxic, SPATE family autotransporter (AT), SigA, which has cytopathic activity towards cultured epithelial cells. Its cytopathic activity is due to its ability to degrade the cytoskeletal protein, α-fodrin. The mechanisms by which AT toxins target cells and tissues differ and the details of how SigA acts are not known. In the current study, the determinants of proteolysis and cell-targeting for SigA were determined. We demonstrate that the SigA passenger or α-domain consists of two functionally distinct domains, designated α1 and α2, which are sufficient to specify proteolytic and cell-binding activities, respectively.

  7. High order sub-cell finite volume schemes for solving hyperbolic conservation laws I: basic formulation and one-dimensional analysis

    Science.gov (United States)

    Pan, JianHua; Ren, YuXin

    2017-08-01

    In this paper, a family of sub-cell finite volume schemes for solving the hyperbolic conservation laws is proposed and analyzed in one-dimensional cases. The basic idea of this method is to subdivide a control volume (main cell) into several sub-cells and the finite volume discretization is applied to each of the sub-cells. The averaged values on the sub-cells of current and face neighboring main cells are used to reconstruct the polynomial distributions of the dependent variables. This method can achieve arbitrarily high order of accuracy using a compact stencil. It is similar to the spectral volume method incorporating with PNPM technique but with fundamental differences. An elaborate utilization of these differences overcomes some shortcomings of the spectral volume method and results in a family of accurate and robust schemes for solving the hyperbolic conservation laws. In this paper, the basic formulation of the proposed method is presented. The Fourier analysis is performed to study the properties of the one-dimensional schemes. A WENO limiter based on the secondary reconstruction is constructed.

  8. Determinants of the renin-angiotensin-aldosterone system in cirrhosis with special emphasis on the central blood volume

    DEFF Research Database (Denmark)

    Møller, Søren; Bendtsen, Flemming; Henriksen, Jens Henrik

    2006-01-01

    OBJECTIVE: Several studies have shown activation of the renin-angiotensin-aldosterone system (RAAS) in cirrhosis. Although the activated RAAS may have several determinants, the system is often considered a surrogate marker of effective hypovolaemia. In this study we investigated the activity...

  9. The number of circulating CD14+ cells is related to infarct size and postinfarct volumes in ST segment elevation myocardial infarction but not non-ST segment elevation myocardial infarction

    Science.gov (United States)

    Montange, Damien; Davani, Siamak; Deschaseaux, Frédéric; Séronde, Marie France; Chopard, Romain; Schiele, François; Jehl, Jérome; Bassand, Jean Pierre; Kantelip, Jean-Pierre; Meneveau, Nicolas

    2012-01-01

    OBJECTIVE: To determine the relationship between the number of CD14+ cells, myocardial infarct (MI) size and left ventricular (LV) volumes in ST segment elevation MI (STEMI) and non-ST segment elevation MI (NSTEMI) patients. METHODS: A total of 62 patients with STEMI (n=34) or NSTEMI (n=28) were enrolled. The number of CD14+ cells was assessed at admission. Infarct size, left ventricular ejection fraction (LVEF) and LV volumes were measured using magnetic resonance imaging five days after MI and six months after MI. Results: In STEMI patients, the number of CD14+ cells was positively and significantly correlated with infarct size at day 5 (r=0.40; P=0.016) and after six months (r=0.34; P=0.047), negatively correlated with LVEF at day 5 (r=−0.50; P=0.002) and after six months (r=−0.46; P=0.005) and positively correlated with end-diastolic (r=0.38; P=0.02) and end-systolic (r=0.49; P=0.002) volumes after six months. In NSTEMI patients, no significant correlation was found between the number of CD14+ cells and infarct size, LVEF or LV volumes at day 5 or after six months. CONCLUSIONS: The number of CD14+ cells at admission was associated with infarct size and LV remodelling in STEMI patients with large infarct size, whereas in NSTEMI patients, no relationship was observed between numbers of CD14+ cells and LV remodelling. PMID:23620701

  10. Disorders of erythrocyte volume homeostasis.

    Science.gov (United States)

    Glogowska, E; Gallagher, P G

    2015-05-01

    Inherited disorders of erythrocyte volume homeostasis are a heterogeneous group of rare disorders with phenotypes ranging from dehydrated to overhydrated erythrocytes. Clinical, laboratory, physiologic, and genetic heterogeneities characterize this group of disorders. A series of recent reports have provided novel insights into our understanding of the genetic bases underlying some of these disorders of red cell volume regulation. This report reviews this progress in understanding determinants that influence erythrocyte hydration and how they have yielded a better understanding of the pathways that influence cellular water and solute homeostasis.

  11. Determinants of acute mortality of Hippodamia convergens (Coleoptera: Coccinellidae) to ultra-low volume permethrin used for mosquito management.

    Science.gov (United States)

    Peterson, Robert K D; Preftakes, Collin J; Bodin, Jennifer L; Brown, Christopher R; Piccolomini, Alyssa M; Schleier, Jerome J

    2016-01-01

    There are relatively few experimental studies and risk assessments of the effects on non-target insects from ultra-low volume (ULV) insecticides used for management of adult mosquitoes. Therefore, we evaluated factors that may influence the ability of an insect to intercept the insecticide at the time of application by using Hippodamia convergens (Coleoptera: Coccinellidae) in field bioassay experiments in 2011 and 2015. Treatment factors included different distances, two cage heights (ground-level and 1.5 m above ground) to the point of the application, and covered vs. uncovered cage faces (2015 only). Insecticides used included a water-based formulation (Aqua-Reslin®) and an oil-based formulation (Permanone® 30-30) of permethrin. Cage height was highly significant both years, with much less acute (i.e., short-term exposure) mortality at ground-level compared with 1.5 m. In 2011, acute mortality was less at ground-level (mean = 3.2%, median = 0%) compared to 1.5 m (mean = 85.2%, median = 100%). Cage type also was highly significant, with less mortality in covered cages compared to uncovered cages. Mortality by cage height and cage type was as follows: ground level, covered cage (mean = 2.8%, median = 0.1%); ground level, uncovered cage (mean = 41.9%, median = 9.6%); 1.5 m, covered cage (mean = 6.8%, median = 0%); 1.5 m, uncovered cage (mean = 83.7%, median = 100%). Results suggest that acute mortality to non-target insects may vary considerably based on their height and their ability to directly intercept the insecticide as the aerosol passes through the area being sprayed.

  12. Comparative method validation for closantel determination in cattle and sheep milk according to European Union Volume 8 and Veterinary International Conference on Harmonization guidelines.

    Science.gov (United States)

    Devreese, Mathias; Maes, An; De Baere, Siegrid; De Backer, Patrick; Croubels, Siska

    2014-08-01

    A specific and sensitive LC-MS/MS method was developed for quantitative determination of closantel in bovine and ovine colostrum and tank milk. Sample preparation consisted of extracting milk samples with acetonitrile/acetone (80/20, v/v) followed by SPE clean-up with Oasis mixed anion exchange columns. After elution with 5% formic acid in acetonitrile and evaporation to dryness, the residue was reconstituted in acetonitrile and water. HPLC separation was achieved on a Zorbax Eclipse Plus C18 column and a gradient elution program with 1mM ammonium acetate in water and acetonitrile. For closantel determination in bovine milk, the method was validated according to EU Volume 8 guidelines whereas for ovine milk both EU Volume 8 and VICH GL49 criteria were applied. The linear range of the method is between 10 and 2000 μg/kg, the limit of quantification 10 μg/kg and limit of detection is 0.63 and 0.32 μg/kg for sheep colostrum and tank milk and 1.27 and 1.24 μg/kg for cattle colostrum and tank milk, respectively. Both guidelines cover a similar set of parameters (linearity, accuracy, precision, limit of detection and limit of quantification), although the acceptance criteria might differ (accuracy and precision) or no specific acceptability ranges are specified in neither guidelines (LOD and LOQ). For some parameters, only one of the guidelines indicates acceptance criteria: EU Volume 8 for applicability, practicability and susceptibility and VICH GL 49 for linearity, specificity and analyte stability. Copyright © 2014 Elsevier B.V. All rights reserved.

  13. Protein segregation between dividing hematopoietic progenitor cells in the determination of the symmetry/asymmetry of cell division.

    Science.gov (United States)

    Nteliopoulos, Georgios; Gordon, Myrtle Y

    2012-09-20

    In the present study, we investigated how the symmetry/asymmetry of cell division in mitotic CD34(+) cells can be evaluated by determining the plane of cell division and the potential distribution of proteins between daughter cells. The orientation of the mitotic spindle is dependent upon the positioning of the centrosomes, which determine the plane of cell division and the sharing of proteins. If the functions of unequally shared proteins are relevant to the kinetics of cell division, they could determine whether the daughter cells undergo self-renewal or differentiation. The kinetic function of the proteins of interest was investigated using a colony-replating assay and carboxyfluorescein succinimidyl ester (CFSE) staining. We used Notch/Numb as a model system, since they have a role in balancing symmetric/asymmetric divisions. Mitotic cells were examined microscopically and centrosomal markers γ-tubulin/pericentrin were used with activated Notch-1 and Numb. We monitored the first crucial divisions by CFSE staining and found an inverse relationship between activated Notch and Numb expression, suggesting a reciprocal regulation. We suggest that the subpopulations expressing activated Notch or Numb have different cell fates. To determine the influence of Notch signaling on progenitor cell self-renewal, we used the γ-secretase inhibitor N-[N-(3,5-Difluorophenacetyl-L-alanyl)]-S-phenylglycine t-Butyl ester (DAPT). DAPT influences self-renewal/differentiation outcome by affecting the frequency of symmetric renewal divisions without affecting the rate of divisions. Overall, the purpose of this study was to establish a cellular system for predicting the symmetry/asymmetry of hematopoietic progenitor divisions at the level of centrosomes and protein distribution and to investigate the influence of these proteins on progenitor cell kinetics.

  14. Impact of medium volume and oxygen concentration in the incubator on pericellular oxygen concentration and differentiation of murine chondrogenic cell culture.

    Science.gov (United States)

    Oze, Hiroki; Hirao, Makoto; Ebina, Kosuke; Shi, Kenrin; Kawato, Yoshitaka; Kaneshiro, Shoichi; Yoshikawa, Hideki; Hashimoto, Jun

    2012-02-01

    Previous studies have demonstrated that oxygen environment is an important determinate factor of cell phenotypes and differentiation, although factors which affect pericellular oxygen concentration (POC) in murine chondrogenic cell culture remain unidentified. Oxygen concentrations in vivo were measured in rabbit musculoskeletal tissues, which were by far hypoxic compared to 20% O(2) (ranging from 2.29 ± 1.16 to 4.36 ± 0.51%). Oxygen concentrations in murine chondrogenic cell (C3H10T1/2) culture medium were monitored in different oxygen concentrations (20% or 5%) in the incubator and in different medium volumes (3,700 or 7,400 μl) within 25-cm(2) flasks. Chondrogenic differentiation was assessed by glycosaminoglycan production with quantitative evaluation of Alcian blue staining in 12-well culture dishes. Expression of chondrogenic genes, aggrecan, and type II collagen α1, was examined by quantitative real-time polymerase chain reaction. Oxygen concentrations in medium decreased accordingly with the depth from medium surface, and POC at Day 6 was 18.99 ± 0.81% in 3,700-μl medium (1,480-μm depth) and 13.26 ± 0.23% in 7,400-μl medium (2,960-μm depth) at 20% O(2) in the incubator, which was 4.96 ± 0.08% (1,480-μm depth) and 2.83 ± 0.42% (2,960-μm depth) at 5% O(2), respectively. The differences of POC compared by medium volume were statistically significant (p = 0.0003 at 20% and p = 0.001 at 5%). Glycosaminoglycan production and aggrecan gene expression were most promoted when cultured in moderately low POC, 1,000 μl (2,960-μm depth) at 20% O(2) and 500 μl (1,480-μm depth) at 5% O(2) in 12-well culture dishes. We demonstrate that medium volume and oxygen concentration in the incubator affect not only POC but also chondrogenic differentiation.

  15. Cell proliferation is a key determinant of the outcome of FOXO3a activation

    Energy Technology Data Exchange (ETDEWEB)

    Poulsen, Raewyn C., E-mail: raewyn.poulsen@gmail.com; Carr, Andrew J.; Hulley, Philippa A.

    2015-06-19

    The FOXO family of forkhead transcription factors have a pivotal role in determining cell fate in response to oxidative stress. FOXO activity can either promote cell survival or induce cell death. Increased FOXO-mediated cell death has been implicated in the pathogenesis of degenerative diseases affecting musculoskeletal tissues. The aim of this study was to determine the conditions under which one member of the FOXO family, FOXO3a, promotes cell survival as opposed to cell death. Treatment of primary human tenocytes with 1 pM hydrogen peroxide for 18 h resulted in increased protein levels of FOXO3a. In peroxide-treated cells cultured in low serum media, FOXO3a inhibited cell proliferation and protected against apoptosis. However in peroxide treated cells cultured in high serum media, cell proliferation was unchanged but level of apoptosis significantly increased. Similarly, in tenocytes transduced to over-express FOXO3a, cell proliferation was inhibited and level of apoptosis unchanged in cells cultured in low serum. However there was a robust increase in cell death in FOXO3a-expressing cells cultured in high serum. Inhibition of cell proliferation in either peroxide-treated or FOXO3a-expressing cells cultured in high serum protected against apoptosis induction. Conversely, addition of a Chk2 inhibitor to peroxide-treated or FOXO3a-expressing cells overrode the inhibitory effect of FOXO3a on cell proliferation and led to increased apoptosis in cells cultured in low serum. This study demonstrates that proliferating cells may be particularly susceptible to the apoptosis-inducing actions of FOXO3a. Inhibition of cell proliferation by FOXO3a may be a critical event in allowing the pro-survival rather than the pro-apoptotic activity of FOXO3a to prevail. - Highlights: • FOXO3a activity can result in either promotion of cell survival or apoptosis. • The outcome of FOXO3a activation differs in proliferating compared to non-proliferating cells. • Proliferating

  16. (99m) Tc-labelled human serum albumin cannot replace (125) I-labelled human serum albumin to determine plasma volume in patients with liver disease

    DEFF Research Database (Denmark)

    Henriksen, Ulrik Lütken; Henriksen, Jens H; Bendtsen, Flemming

    2013-01-01

    -labelled human serum albumin (99mTc-HSA) and iodine-labelled human serum albumin (125I-HSA), as the former may have advantages at repeated measurements and the latter is the classical gold standard. Study population and methods In 88 patients, (64 with liver disease, mainly cirrhosis, and 24 patients without......Summary Background and aims Determination of plasma volume (PV) is important in several clinical situations. Thus, patients with liver disease often have augmented PV as part of their sodium–water retention. This study was undertaken to compare PV determination by two indicators: technetium...... liver disease), simultaneous measurements of PV were taken with 99mTc-HSA and 125I-HSA after 1 h in the supine position. Blood samples were obtained before and 10 min after quantitative injection of the two indicators. In a subset of patients (n = 32), the measurements were repeated within 1 h. Results...

  17. Determination of fugitive coal-dust emissions from rotary railcar dumping. Volume 2. Appendices A-H

    Energy Technology Data Exchange (ETDEWEB)

    Brookman, E.T.; Carnes, D.H.; Catizone, P.A.; Kelley, K.J.

    1984-09-01

    TRC Environmental Consultants, Inc. conducted 72 field tests to determine particulate emission levels for a rotary railcar coal dumper operated by the Potomac Electric Power Company. Approximately 60 parameters were monitored as part of the testing program. These parameters related to particulate emissions, meteorology, and coal properties. The railcar dumper was situated inside of a shed. No other control devices were in operation during the test program. Emissions were measured at the entrance and exit doorways of the shed using the mass flux technique. Background levels were accounted for using the upwind/downwind technique. The amount of particulate material that deposited within the shed was also recorded. The study resulted in the determination of emission levels that were one to two orders of magnitude less than those cited in the literature. These emission levels were found to correlate the strongest with the moisture content of the coal and whether the coal was washed or unwashed.

  18. Determinação da curva volume versus duração por meio da distribuição do valor extremo generalizada para estimativa do volume de espera em reservatórios = Determination of volume duration curve by generalized extreme value distribution to estimate waiting water volume in reservoirs

    Directory of Open Access Journals (Sweden)

    2007-01-01

    Full Text Available Nos reservatórios de usos múltiplo de água, existe conflito para o controle de cheias, principalmente para a geração de energia elétrica. Para controlar cheias é necessário um volume vazio capaz de absorver uma eventual cheia sem causar danos nas áreas à jusante. Por outro lado, para gerar energia é desejável alocar o menor volume de proteção possível, ao passo que para controlar cheias é desejável ter uma estimativa confiável da possibilidade do reservatório falhar quando um dado volume de proteção é alocado. O objetivo deste trabalho foi construir a Curva Volume x Duração, empregada para estimar volume de espera, através do ajuste das séries de volumes máximos afluentes usando a distribuição GEV e momentos LH. Foram usados volumes afluentes observados em várias estações fluviométricas de rios do estado do Paraná, Brasil. Todos os ajustes dos volumesmáximos afluentes foram aceitos através do teste de qualidade de ajuste proposto por Wang (1998. A curva volume x duração foi adequadamente construída usando valores de volumes máximos afluentes observados em diferentes estações fluviométricas.In the reservoirs of multiple uses of water, exist conflict to control floods, mainly for generation of electric energy. To control floods it's necessary an empty volume able to absorb an accidental flood without causing damage in the areas downstream. On the other side, to generate energy it's desirable to place the smallest volume of possible protection, whereas to control floods it's desirable to have a reliable estimate in case the reservoir fails when a given volume of protection is placed. The objective of this work was to construct the Volume Curve x Length, used to estimate the waiting volume, through the adjustments of the series of the affluent maximum volume using the distribution GEV and moments LH. Affluent volumeswere used and observed in several fluviometrics stations of rivers in Paraná state, Brazil

  19. 脂质体包封体积测定方法的研究进展%Progress on the determination of liposome encapsulated volume

    Institute of Scientific and Technical Information of China (English)

    隋月; 宋艳志; 邓意辉

    2013-01-01

    Objective To review the methods on the determination of liposome encapsulated volume.Methods The review was summarized on the basis of 57 published literatures.Results Based on different principles,the determination of liposome encapsulated volume was divided into two types:theoretical model and experimental determination.Experimental determination included solute encapsulation method,electron spin resonance (ESR) method,conductivity and nuclear magnetic resonance (NMR) method.Conclusions The article provides an important reference for research and development of encapsulated water-soluble,nucleic acid and protein drugs in the aqueous phase of the liposome.%目的 对脂质体包封体积的测定方法进行综述.方法 查阅国内外相关文献57篇,对包封体积的测定方法进行归纳、总结和分析.结果 基于不同原理,包封体积的测定方法分为理论模型和实验测定两类,其中,实验测定包括溶质包封(solute encapsulation)、电子自旋共振(electron spin resonance,ESR)、电导率(conductivity)及核磁共振(nuclear magnetic resonance,NMR)法.结论 为进一步研究脂质体内水相中包封的水溶性、核酸和蛋白质类药物提供重要参考.

  20. Determining the cellular diversity of hepatitis C virus quasispecies by single-cell viral sequencing.

    Science.gov (United States)

    McWilliam Leitch, E Carol; McLauchlan, John

    2013-12-01

    Single-cell genomics is emerging as an important tool in cellular biology. We describe for the first time a system to investigate RNA virus quasispecies diversity at the cellular level utilizing hepatitis C virus (HCV) replicons. A high-fidelity nested reverse transcription (RT)-PCR assay was developed, and validation using control transcripts of known copy number indicated a detection limit of 3 copies of viral RNA/reaction. This system was used to determine the cellular diversity of subgenomic JFH-1 HCV replicons constitutively expressed in Huh7 cells. Each cell contained a unique quasispecies that was much less diverse than the quasispecies of the bulk cell population from which the single cells were derived, suggesting the occurrence of independent evolution at the cellular level. An assessment of the replicative fitness of the predominant single-cell quasispecies variants indicated a modest reduction in fitness compared to the wild type. Real-time RT-PCR methods capable of determining single-cell viral loads were developed and indicated an average of 113 copies of replicon RNA per cell, correlating with calculated RNA copy numbers in the bulk cell population. This study introduces a single-cell RNA viral-sequencing method with numerous potential applications to explore host-virus interactions during infection. HCV quasispecies diversity varied greatly between cells in vitro, suggesting different within-cell evolutionary pathways. Such divergent trajectories in vivo could have implications for the evolution and establishment of antiviral-resistant variants and host immune escape mutants.

  1. ROS-induced autophagy in cancer cells assists in evasion from determinants of immunogenic cell death

    NARCIS (Netherlands)

    Garg, A.D.; Dudek, A.M.D.; Ferreira, G.B.; Verfaillie, T.; Vandenabeele, P.; Krysko, D.V.; Mathieu, C.; Agostinis, P.

    2013-01-01

    Calreticulin surface exposure (ecto-CALR), ATP secretion, maturation of dendritic cells (DCs) and stimulation of T cells are prerequisites for anticancer therapy-induced immunogenic cell death (ICD). Recent evidence suggests that chemotherapy-induced autophagy may positively regulate ICD by favoring

  2. Volume Regulated Channels

    DEFF Research Database (Denmark)

    Klausen, Thomas Kjær

    - serves a multitude of functions in the mammalian cell, regulating the membrane potential (Em), cell volume, protein activity and the driving force for facilitated transporters giving Cl- and Cl- channels a major potential of regulating cellular function. These functions include control of the cell cycle...... of volume perturbations evolution have developed system of channels and transporters to tightly control volume homeostasis. In the past decades evidence has been mounting, that the importance of these volume regulated channels and transporters are not restricted to the defense of cellular volume......, controlled cell death and cellular migration. Volume regulatory mechanisms has long been in focus for regulating cellular proliferation and my thesis work have been focusing on the role of Cl- channels in proliferation with specific emphasis on ICl, swell. Pharmacological blockage of the ubiquitously...

  3. Testosterone level, nasal gland volume and Leydig cell morphometry in capybaras (Hydrochoerus hydrochaeris Níveis de testosterona, volume da glândula nasal e morfometria das células de Leydig em capivaras (Hydrochoerus hydrochaeris

    Directory of Open Access Journals (Sweden)

    D.S. Costa

    2006-12-01

    Full Text Available The testosterone levels with nasal gland volume and Leydig cell morphometry in sexually mature capybaras were compared using 10 adult capybaras, aging 28 months. Body and testicular weight were 50kg and 28g, respectively. The gonadosomatic index was 0.11±0.02%, while nasal gland volume was 34.12±13.04ml. Serum testosterone concentration was 25.07±14.73ng/ml. Leydig cell volume density and leydigosomatic index were, respectively, 28.8±8.3% and 0.030±0.007%. Total and individual Leydig cell volumes were 6.88±1.92ml and 2,228.14±272.59x10-12ml, respectively. Leydig cell numbers per testis and per gram of testis were, respectively, 3,079.4±719.9x10(6 and 115.09±31.78x10(6 cells. These results show that increase in serum testosterone concentration is correlated to a proportional increase of nasal gland and Leydig cell volume in capybaras.Estimaram-se as correlações entre os níveis de testosterona e o volume da glândula nasal e a morfometria das células de Leydig no parênquima testicular de capivaras sexualmente maduras. Utilizaram-se 10 capivaras com aproximadamente 28 meses de idade e com peso médio corporal de 50kg e testicular de 28g. O índice gonadossomático e o volume médio da glândula nasal foram 0,11±0,02% e 34,12±13,04ml, respectivamente, e a concentração sérica de testosterona 25,07±14,73ng/ml. A proporção volumétrica de células de Leydig e o índice leydigossomático foram 28,8± 8,3% e 0,030±0,007%, respectivamente. Os volumes total e individual de células de Leydig foram 6,88±1,92ml e 2228,14±272,59´10-12ml, respectivamente. O número de células de Leydig por testículo e por grama de testículo foram, respectivamente, 3079,4±719,9´10(6 e 115,09±31,78´10(6. Concluiu-se que o aumento dos níveis séricos de testosterona foi acompanhado por aumento proporcional do volume da glândula nasal de capivaras e que a concentração desse hormônio é tanto mais alta quanto maior for o volume da célula de

  4. Right ventricular volume and mass determined by cine magnetic resonance imaging in HIV patients with possible right ventricular dysfunction

    DEFF Research Database (Denmark)

    Kjaer, Andreas; Lebech, Anne-Mette; Gerstoft, Jan

    2006-01-01

    Impaired right ventricular (RV) function has been reported to occur in patients with HIV when studied by echocardiography. However, for accurate evaluation of RV function and morphology, first-pass radionuclide ventriculography (RNV) and cine magnetic resonance imaging (cine-MRI) are methods...... ventricular ejection fraction (RVEF). To do so, we screened patients with RNV and performed an additional cine-MRI in those with reduced RVEF determined by RNV. Ninety patients with HIV were included. To evaluate the MRI measures exactly we included 18 age- and gender-matched healthy volunteers to establish...

  5. Right ventricular volume and mass determined by cine magnetic resonance imaging in HIV patients with possible right ventricular dysfunction

    DEFF Research Database (Denmark)

    Kjaer, Andreas; Lebech, Anne-Mette; Gerstoft, Jan

    2006-01-01

    Impaired right ventricular (RV) function has been reported to occur in patients with HIV when studied by echocardiography. However, for accurate evaluation of RV function and morphology, first-pass radionuclide ventriculography (RNV) and cine magnetic resonance imaging (cine-MRI) are methods...... ventricular ejection fraction (RVEF). To do so, we screened patients with RNV and performed an additional cine-MRI in those with reduced RVEF determined by RNV. Ninety patients with HIV were included. To evaluate the MRI measures exactly we included 18 age- and gender-matched healthy volunteers to establish...

  6. Correlation between [{sup 18}F]FDG PET/CT and volume perfusion CT in primary tumours and mediastinal lymph nodes of non-small-cell lung cancer

    Energy Technology Data Exchange (ETDEWEB)

    Sauter, Alexander W.; Spira, Daniel; Schulze, Maximilian; Pfannenberg, Christina; Claussen, Claus D.; Horger, Marius S. [Eberhard Karls University, Diagnostic and Interventional Radiology, Department of Radiology, Tuebingen (Germany); Hetzel, Juergen [Eberhard Karls University, Department of Oncology, Hematology, Immunology, Rheumatology and Pulmonology, Tuebingen (Germany); Reimold, Matthias [Eberhard Karls University, Nuclear Medicine, Department of Radiology, Tuebingen (Germany); Klotz, Ernst [Siemens Healthcare, Computed Tomography, Forchheim (Germany)

    2013-05-15

    The aim of this study was to investigate correlations between glucose metabolism as determined by [{sup 18}F]FDG PET/CT and tumour perfusion as quantified by volume perfusion CT in primary tumours and mediastinal lymph nodes (MLN) of patients with non-small-cell lung cancer (NSCLC). Enrolled in the study were 17 patients with NSCLC. [{sup 18}F]FDG uptake was quantified in terms of SUV{sub max} and SUV{sub avg}. Blood flow (BF), blood volume (BV) and flow extraction product (K{sup trans}) were determined as perfusion parameters. The correlations between the perfusion parameters and [{sup 18}F]FDG uptake values were subsequently evaluated. For the primary tumours, no correlations were found between perfusion parameters and [{sup 18}F]FDG uptake. In MLN, there were negative correlations between BF and SUV{sub avg} (r = -0.383), BV and SUV{sub avg} (r = -0.406), and BV and SUV{sub max} (r = -0.377), but not between BF and SUV{sub max}, K{sup trans} and SUV{sub avg}, or K{sup trans} and SUV{sub max}. Additionally, in MLN with SUV{sub max} >2.5 there were negative correlations between BF and SUV{sub avg} (r = -0.510), BV and SUV{sub avg} (r = -0.390), BF and SUV{sub max} (r = -0.536), as well as BV and SUV{sub max} (r = -0.346). Perfusion and glucose metabolism seemed to be uncoupled in large primary tumours, but an inverse correlation was observed in MLN. This information may help improve therapy planning and response evaluation. (orig.)

  7. Circuits and methods for determination and control of signal transition rates in electrochemical cells

    Science.gov (United States)

    Jamison, David Kay

    2016-04-12

    A charge/discharge input is for respectively supplying charge to, or drawing charge from, an electrochemical cell. A transition modifying circuit is coupled between the charge/discharge input and a terminal of the electrochemical cell and includes at least one of an inductive constituent, a capacitive constituent and a resistive constituent selected to generate an adjusted transition rate on the terminal sufficient to reduce degradation of a charge capacity characteristic of the electrochemical cell. A method determines characteristics of the transition modifying circuit. A degradation characteristic of the electrochemical cell is analyzed relative to a transition rate of the charge/discharge input applied to the electrochemical cell. An adjusted transition rate is determined for a signal to be applied to the electrochemical cell that will reduce the degradation characteristic. At least one of an inductance, a capacitance, and a resistance is selected for the transition modifying circuit to achieve the adjusted transition rate.

  8. Cell number and transfection volume dependent peptide nucleic acid antisense activity by cationic delivery methods

    DEFF Research Database (Denmark)

    Llovera Nadal, Laia; Berthold, Peter; Nielsen, Peter E

    2012-01-01

    Efficient intracellular delivery is essential for high activity of nucleic acids based therapeutics, including antisense agents. Several strategies have been developed and practically all rely on auxiliary transfection reagents such as cationic lipids, cationic polymers and cell penetrating...... peptides as complexing agents and carriers of the nucleic acids. However, uptake mechanisms remain rather poorly understood, and protocols always require optimization of transfection parameters. Considering that cationic transfection complexes bind to and thus may up-concentrate on the cell surface, we...... have now quantitatively compared the cellular activity (in the pLuc705 HeLa cell splice correction system) of PNA antisense oligomers using lipoplex delivery of cholesterol- and bisphosphonate-PNA conjugates, polyplex delivery via a PNA-polyethyleneimine conjugate and CPP delivery via a PNA...

  9. Large volume unresectable locally advanced non-small cell lung cancer: acute toxicity and initial outcome results with rapid arc

    Directory of Open Access Journals (Sweden)

    Fogliata Antonella

    2010-10-01

    Full Text Available Abstract Background To report acute toxicity, initial outcome results and planning therapeutic parameters in radiation treatment of advanced lung cancer (stage III with volumetric modulated arcs using RapidArc (RA. Methods Twenty-four consecutive patients were treated with RA. All showed locally advanced non-small cell lung cancer with stage IIIA-IIIB and with large volumes (GTV:299 ± 175 cm3, PTV:818 ± 206 cm3. Dose prescription was 66Gy in 33 fractions to mean PTV. Delivery was performed with two partial arcs with a 6 MV photon beam. Results From a dosimetric point of view, RA allowed us to respect most planning objectives on target volumes and organs at risk. In particular: for GTV D1% = 105.6 ± 1.7%, D99% = 96.7 ± 1.8%, D5%-D95% = 6.3 ± 1.4%; contra-lateral lung mean dose resulted in 13.7 ± 3.9Gy, for spinal cord D1% = 39.5 ± 4.0Gy, for heart V45Gy = 9.0 ± 7.0Gy, for esophagus D1% = 67.4 ± 2.2Gy. Delivery time was 133 ± 7s. At three months partial remission > 50% was observed in 56% of patients. Acute toxicities at 3 months showed 91% with grade 1 and 9% with grade 2 esophageal toxicity; 18% presented grade 1 and 9% with grade 2 pneumonia; no grade 3 acute toxicity was observed. The short follow-up does not allow assessment of local control and progression free survival. Conclusions RA proved to be a safe and advantageous treatment modality for NSCLC with large volumes. Long term observation of patients is needed to assess outcome and late toxicity.

  10. Volume of white matter hyperintensities is an independent predictor of intelligence quotient and processing speed in children with sickle cell disease.

    Science.gov (United States)

    van der Land, Veronica; Hijmans, Channa T; de Ruiter, Marieke; Mutsaerts, Henri J M M; Cnossen, Marjon H; Engelen, Marc; Majoie, Charles B L M; Nederveen, Aart J; Grootenhuis, Martha A; Fijnvandraat, Karin

    2015-02-01

    Sickle cell disease can be complicated by cerebral white matter hyperintensities (WMHs), which are associated with diminished neurocognitive functioning. The influence of the total volume of WMHs on the degree of neurocognitive dysfunction has not yet been characterized. In our study of 38 patients (mean age 12·5 years) we demonstrated that a higher volume of WMHs was associated with lower full-scale intelligence quotient (IQ), verbal IQ, Processing Speed Index and more fatigue. Our results suggest that volume of WMHs is an additional parameter to take into account when planning individual diagnostic and treatment options. © 2014 John Wiley & Sons Ltd.

  11. Sample Size Determination: A Comparison of Attribute, Continuous Variable, and Cell Size Methods.

    Science.gov (United States)

    Clark, Philip M.

    1984-01-01

    Describes three methods of sample size determination, each having its use in investigation of social science problems: Attribute method; Continuous Variable method; Galtung's Cell Size method. Statistical generalization, benefits of cell size method (ease of use, trivariate analysis and trichotyomized variables), and choice of method are…

  12. Discovering naturally processed antigenic determinants that confer protective T cell immunity

    DEFF Research Database (Denmark)

    Gilchuk, Pavlo; Spencer, Charles T; Conant, Stephanie B;

    2013-01-01

    CD8+ T cells (TCD8) confer protective immunity against many infectious diseases, suggesting that microbial TCD8 determinants are promising vaccine targets. Nevertheless, current T cell antigen identification approaches do not discern which epitopes drive protective immunity during active infectio...

  13. Local 3D matrix confinement determines division axis through cell shape.

    Science.gov (United States)

    He, Lijuan; Chen, Weitong; Wu, Pei-Hsun; Jimenez, Angela; Wong, Bin Sheng; San, Angela; Konstantopoulos, Konstantinos; Wirtz, Denis

    2016-02-09

    How the division axis is determined in mammalian cells embedded in three-dimensional (3D) matrices remains elusive, despite that many types of cells divide in 3D environments. Cells on two-dimensional (2D) substrates typically round up completely to divide. Here, we show that in 3D collagen matrices, mammalian cells such as HT1080 human fibrosarcoma and MDA-MB-231 breast cancer cells exhibit division modes distinct from their Counterparts on 2D substrates, with a markedly higher fraction of cells remaining highly elongated through mitosis in 3D matrices. The long axis of elongated mitotic cells accurately predicts the division axis, independently of matrix density and cell-matrix interactions. This 3D-specific elongated division mode is determined by the local confinement produced by the matrix and the ability of cells to protrude and locally remodel the matrix via β1 integrin. Elongated division is readily recapitulated using collagen-coated microfabricated channels. Cells depleted of β1 integrin still divide in the elongated mode in microchannels, suggesting that 3D confinement is sufficient to induce the elongated cell-division phenotype.

  14. Principles of Bacterial Cell-Size Determination Revealed by Cell-Wall Synthesis Perturbations

    Directory of Open Access Journals (Sweden)

    Carolina Tropini

    2014-11-01

    Full Text Available Although bacterial cell morphology is tightly controlled, the principles of size regulation remain elusive. In Escherichia coli, perturbation of cell-wall synthesis often results in similar morphologies, making it difficult to deconvolve the complex genotype-phenotype relationships underlying morphogenesis. Here we modulated cell width through heterologous expression of sequences encoding the essential enzyme PBP2 and through sublethal treatments with drugs that inhibit PBP2 and the MreB cytoskeleton. We quantified the biochemical and biophysical properties of the cell wall across a wide range of cell sizes. We find that, although cell-wall chemical composition is unaltered, MreB dynamics, cell twisting, and cellular mechanics exhibit systematic large-scale changes consistent with altered chirality and a more isotropic cell wall. This multiscale analysis enabled identification of distinct roles for MreB and PBP2, despite having similar morphological effects when depleted. Altogether, our results highlight the robustness of cell-wall synthesis and physical principles dictating cell-size control.

  15. Closed system generation of dendritic cells from a single blood volume for clinical application in immunotherapy

    NARCIS (Netherlands)

    Elias, M; van Zanten, J; Hospers, GAP; Setroikromo, A; de Jong, MA; de Leij, LFMH; Mulder, NH

    2005-01-01

    Dendritic cells (DC) used for clinical trials should be processed oil a large scale conforming to current good manufacturing practice (cGM P) guidelines. The aim of this study was to develop a protocol for clinical grade generation of immature DC in a closed-systern. Aphereses were performed with th

  16. Observations from using models to fit the gas production of varying volume test cells and landfills.

    Science.gov (United States)

    Lamborn, Julia

    2012-12-01

    Landfill operators are looking for more accurate models to predict waste degradation and landfill gas production. The simple microbial growth and decay models, whilst being easy to use, have been shown to be inaccurate. Many of the newer and more complex (component) models are highly parameter hungry and many of the required parameters have not been collected or measured at full-scale landfills. This paper compares the results of using different models (LANDGEM, HBM, and two Monod models developed by the author) to fit the gas production of laboratory scale, field test cell and full-scale landfills and discusses some observations that can be made regarding the scalability of gas generation rates. The comparison of these results show that the fast degradation rate that occurs at laboratory scale is not replicated at field-test cell and full-scale landfills. At small scale, all the models predict a slower rate of gas generation than actually occurs. At field test cell and full-scale a number of models predict a faster gas generation than actually occurs. Areas for future work have been identified, which include investigations into the capture efficiency of gas extraction systems and into the parameter sensitivity and identification of the critical parameters for field-test cell and full-scale landfill predication.

  17. The Relationship between Erythrocyte Volume and Cell Age in Humans and Baboons.

    Science.gov (United States)

    1983-03-30

    IM weekly to prevent reutilization of s9Fe. Red cell ....................... .F. -7 -J %J W. J W.I 7A 01 fractionation into size-dependent...management of thalassemia . Blood 55:55, 1980. 12. Corash L, Klein H, Deisseroth A, Shafer B, Rosen S, Berron J, Griffith P, Nienhuis A: Selective isolation

  18. Stereological estimates of nuclear volume in squamous cell carcinoma of the uterine cervix and its precursors

    DEFF Research Database (Denmark)

    Sørensen, Flemming Brandt; Bichel, P; Jakobsen, A

    1991-01-01

    -sampling of nuclear intercepts in 51 pre-treatment biopsies from patients with invasive squamous cell carcinomas (SCC). Vertical sections from 27 specimens with cervical intraepithelial neoplasia (CIN) grades I through III were also investigated, along with 10 CIN III associated with microinvasion (CIN III + M...

  19. Canola Oil Fuel Cell Demonstration: Volume 3 - Technical, Commercialization, and Application Issues Associated with Harvested Biomass

    Science.gov (United States)

    2006-08-17

    Fuels for Fuel Cells.” International Journal of Hydrogen Energy , vol 26, pp. 291-301. Arthur D. Little, Inc. 2001. Conceptual Design of POX / SOFC 5...Compact Plasmatron-Boosted Hydrogen Generation Technology for Vehicular Applications.” International Journal of Hydrogen Energy , No. 24. pp. 341

  20. Cost of fuel cell systems on a mass basis as a function of production volume; Kosten von Brennstoffzellensystemen auf Massenbasis in Abhaengigkeit von der Absatzmenge

    Energy Technology Data Exchange (ETDEWEB)

    Werhahn, Johannes

    2009-07-01

    The currently high cost of fuel cells is determined by expensive materials and low production volume. A detailed understanding of the cost structures reveals unexploited potential that can reduce costs in future. However, this requires a method of predicting costs that can be applied with little effort and which offers both a sufficient degree of detail and also good accuracy. Existing forecasting methods do not, however, fulfil these requirements. The major objective of the present work was to apply mass-specific cost forecasting to fuel cell systems for the first time and to modify the approach for this application. In this method, the cost of an object is estimated solely by means of the object mass with the aid of empirical values (Euro/kg). The advantages of the method are its simple application and the accuracy of the forecast. Due to the considerable complexity of the fuel cell and the heterogeneity of the materials used, the application of mass-specific cost forecasting does not provide the desired benefits on the level of the aggregated system. The mass-specific cost forecast approach was therefore expanded and optimized. Instead of determining costs on the level of the aggregated system, the cost forecast was applied directly to the individual components. Cost parameters were also embedded in the method in order to include component-internal cost-relevant differences. Due to the great influence of the production rate on the manufacturing costs, an additional dependence on number of units was also integrated. Expanding the empirical values from discrete values to distribution functions enabled a detailed error analysis to be performed and also a statistical localization of the predicted production costs. Empirical values are necessary in order to implement the modified method and therefore an extensive data search was performed. To this end, a methodology was developed which comprehensively described the data acquisition and the required data evaluation on

  1. Pretherapy metabolic tumour volume is an independent predictor of outcome in patients with diffuse large B-cell lymphoma

    Energy Technology Data Exchange (ETDEWEB)

    Sasanelli, Myriam; Meignan, Michel; Haioun, Corinne; Itti, Emmanuel [Paris-Est University, Nuclear Medicine and Lymphoid Malignancies Unit, Henri Mondor Hospital, Creteil (France); Berriolo-Riedinger, Alina; Casasnovas, Rene-Olivier [Nuclear Medicine and Hematology, Georges-Francois Leclerc Center, Le Bocage Hospital, Dijon (France); Biggi, Alberto; Gallamini, Andrea [Nuclear Medicine and Hematology, Santa Croce e Carle Hospital, Cuneo (Italy); Siegel, Barry A.; Cashen, Amanda F. [Washington University School of Medicine, Nuclear Medicine and Oncology, Siteman Cancer Center, St. Louis, MO (United States); Vera, Pierre; Tilly, Herve [Nuclear Medicine and Hematology, Henri Becquerel Center, Rouen (France); Versari, Annibale [Nuclear Medicine, Santa Maria Nuova Hospital-IRCCS, Reggio Emilia (Italy)

    2014-11-15

    We investigated the prognostic value of total metabolic tumour volume (TMTV) in diffuse large B-cell lymphoma (DLBCL). TMTV was measured in 114 patients with newly diagnosed DLBCL who underwent {sup 18}F-FDG PET/CT at baseline before immunochemotherapy. TMTV was computed by summing the volumes of all lymphomatous lesions after applying the local SUVmax threshold of 41 % using semiautomatic software. Prognostic value was assessed by Kaplan-Meier estimates of progression-free survival (PFS) and overall survival (OS). Median follow-up was 39 months. Average pretherapy TMTV was 509 ± 568 cm{sup 3}. The 3-year estimates of PFS were 77 % in the low metabolic burden group (TMTV ≤550 cm{sup 3}) and 60 % in the high metabolic burden group (TMTV >550 cm{sup 3}, p = 0.04), and prediction of OS was even better (87 % vs. 60 %, p = 0.0003). Cox regression showed independence of TMTV for OS prediction (p = 0.002) compared with other pretherapy indices of tumour burden, such as tumour bulk and the International Prognostic Index. Pretherapy TMTV is an independent predictor of outcome in patients with DLBCL. (orig.)

  2. [Metastatic characteristics of lymph node in supraclavicular zone and radiotherapy target volume for limited-stage small cell lung cancer].

    Science.gov (United States)

    Feng, Zhen-xing; Zhao, Lu-jun; Guan, Yong; Sun, Yao; Ji, Kai; Wang, Ping

    2013-05-21

    To explore the reasonable radiotherapy range by analyzing the characteristics of supraclavicular lymph node metastasis in limited-stage small cell lung cancer (LS-SCLC). From January 2005 to December 2011, patients of LS-SCLC were reviewed. Supraclavicular zone was further divided into five subgroups including para-recurrent laryngeal nerve (region I and region II ), para-internal jugular vein (region III ), supraclavicular region (region IV), as well as the other regions except for the mentioned above (region V). The characteristics of the lymph nodes in each region were analyzed. The supraclavicular lymph node metastasis was found in 60 patients, with a positive rate of 34.5%. In multivariate Logistic regression analysis,intra-thoracic lymph node metastasis in the lymph node stations of level 2 and 3 were found to be the risk factors of supraclavicular lymph node metastasis (P = 0.006,P = 0.000). Our data suggests that the frequencies of metastasis in region I and III were much higher than those in the other areas.Among the sixty patients with supraclavicular lymph node metastasis, 95.0% were found at region I or III while the incidence of skip metastasis was only 5.0%. It is advisable to contain the bilateral supraclavicular nodes in patients with mediastinal lymph nodes metastasis to the level 2 or 3 for elective radiation target volume.The clinical target volume (CTV) exterior margin containing the outer margin of internal jugular vein may be suitable.

  3. Kidney volume correlates with tumor diameter in renal cell carcinoma and is associated with histological poor prognostic features.

    Science.gov (United States)

    Hayes, Brian D; Finn, Stephen P

    2014-02-01

    We aimed to correlate kidney volume (KV) in renal cell carcinoma nephrectomy specimens with tumor diameter (TD), macroscopic growth pattern, and histological features associated with poor prognosis. Histopathology reports, macroscopic specimen photographs, and selected glass slides were retrospectively reviewed. KV was approximated to the volume of an ellipsoid. A total of 273 specimens were identified with median KV 245 cm(3). Kidneys larger than this contained larger tumors (7.5 vs 4.5 cm). KV was significantly greater in tumors of high grade, involving perinephric fat, exhibiting venous invasion, and involving renal sinus. There was a robust linear correlation between KV and TD (r = 0.602) and a weaker correlation between kidney diameter (KD) and TD (r = 0.53). In pT1 tumors, KV (r = 0.40) also correlated better with TD than did KD (r = 0.27). By multiple regression analysis, both TD and venous invasion independently predicted both KD (R (2) = 38.27%) and KV (R (2) = 51.97%). KV and KD correlate well with TD and histopathological features of aggressiveness, although KD correlates better overall and in the pT1 subset.

  4. Accurate determination of plasmid copy number of flow-sorted cells using droplet digital PCR.

    Science.gov (United States)

    Jahn, Michael; Vorpahl, Carsten; Türkowsky, Dominique; Lindmeyer, Martin; Bühler, Bruno; Harms, Hauke; Müller, Susann

    2014-06-17

    Many biotechnological processes rely on the expression of a plasmid-based target gene. A constant and sufficient number of plasmids per cell is desired for efficient protein production. To date, only a few methods for the determination of plasmid copy number (PCN) are available, and most of them average the PCN of total populations disregarding heterogeneous distributions. Here, we utilize the highly precise quantification of DNA molecules by droplet digital PCR (ddPCR) and combine it with cell sorting using flow cytometry. A duplex PCR assay was set up requiring only 1000 sorted cells for precise determination of PCN. The robustness of this method was proven by thorough optimization of cell sorting, cell disruption, and PCR conditions. When non plasmid-harboring cells of Pseudomonas putida KT2440 were spiked with different dilutions of the expression plasmid pA-EGFP_B, a PCN from 1 to 64 could be accurately detected. As a proof of principle, induced cultures of P. putida KT2440 producing an EGFP-fused model protein by means of the plasmid pA-EGFP_B were investigated by flow cytometry and showed two distinct subpopulations, fluorescent and nonfluorescent cells. These two subpopulations were sorted for PCN determination with ddPCR. A remarkably diverging plasmid distribution was found within the population, with nonfluorescent cells showing a much lower PCN (≤1) than fluorescent cells (PCN of up to 5) under standard conditions.

  5. Human bone marrow cell culture: a sensitive method for determination of the biocompatibility of implant materials.

    Science.gov (United States)

    Wilke, A; Landgraff, M; Orth, J; Poenitz, H; Kienapfel, H; Boelte, K; Griss, P; Franke, R P

    1999-01-01

    The objective of this study was to develop a test method for determining the cytotoxicity and biocompatibility of various biomaterials that are used in orthopaedic surgery. This method is based on the use of a human bone marrow cell culture and was developed as an alternative to animal experiments. Human bone marrow cell culture has certain advantages over other cell culture models, as its results show a greater conformity with animal experimental results and clinical studies. Primary cell adherence, cell number, cell proliferation, production of extracellular matrix, cell viability and cell differentiation were used as indicative parameters of biocompatibility. After 2 weeks in culture, differences could be observed between the biomaterials with respect to these parameters. Cell numbers were greatest on the hydroxyapatite ceramic specimens, but were decreased on the titanium alloy specimens. Extracellular matrix hydroxyapatite production was high for ceramics, but reduced for titanium specimens. The polymers allowed only a few cells to adhere, and there were no signs of extracellular matrix production. The influence of biomaterials on differentiation of large numbers of cells was analysed by using flow cytophotometry. There were similar populations of T cells and monocytes on all specimens. However, extended B cell and granulocyte populations were observed with titanium and polyethylene.

  6. Large volume of water samples introduced in dispersive liquid-liquid microextraction for the determination of 15 triazole fungicides by gas chromatography-tandem mass spectrometry.

    Science.gov (United States)

    Nie, Jing; Chen, Fujiang; Song, Zhiyu; Sun, Caixia; Li, Zuguang; Liu, Wenhan; Lee, Mawrong

    2016-10-01

    A novel method of large volume of water samples directly introduced in dispersive liquid-liquid microextraction was developed, which is based on ultrasound/manual shaking-synergy-assisted emulsification and self-generating carbon dioxide gas (CO2) breaking down the emulsion for the determination of 15 triazole fungicides by gas chromatography-tandem mass spectrometry. This technique makes low-density extraction solvent toluene (180 μL) dissolve in 200 mL of samples containing 0.05 mol L(-1) of HCl and 5 % of NaCl (w/v) to form a well emulsion by synergy of ultrasound and manual shaking, and injects NaHCO3 solution (1.0 mol L(-1)) to generate CO2 achieving phase separation with the assistance of ultrasound. The entire process is accomplished within 8 min. The injection of NaHCO3 to generate CO2 achieves phase separation that breaks through the centrifugation limited large volume aqueous samples. In addition, the device could be easily cleaned, and this kind of vessel could be reconfigured for any volume of samples. Under optimal conditions, the low limits of detection ranging from 0.7 to 51.7 ng L(-1), wide linearity, and enrichment factors obtained were in the range 924-3669 for different triazole fungicides. Southern end of the Beijing-Hangzhou Grand Canal water (Hangzhou, China) was used to verify the applicability of the developed method. Graphical Abstract Flow chart of ultrasound/manual shaking-synergy-assisted emulsification and self-generating carbon dioxide gas breaking down the emulsion.

  7. The gametic central cell of Arabidopsis determines the lifespan of adjacent accessory cells

    OpenAIRE

    Kägi, Christina; Baumann, Nadine; Nielsen, Nicola; Stierhof, York-Dieter; Groß-Hardt, Rita

    2010-01-01

    Plant germ cells develop in specialized haploid structures, termed gametophytes. The female gametophyte patterns of flowering plants are diverse, with often unknown adaptive value. Here we present the Arabidopsis fiona mutant, which forms a female gametophyte that is structurally and functionally reminiscent of a phylogenetic distant female gametophyte. The respective changes include a modified reproductive behavior of one of the female germ cells (central cell) and an extended lifespan of th...

  8. Development of a mathematical model for a single alkaline membrane fuel cell (AMFC) with fixed volume and general square section

    Energy Technology Data Exchange (ETDEWEB)

    Sommer, Elise Meister; Vargas, Jose Viriato Coelho [Universidade Federal do Parana (UFPR), Curitiba, PR (Brazil). Centro Politecnico. Setor de Tecnologia], Email: jvargas@demec.ufpr.br; Martins, Lauber de Souza; Ordonez, Juan Carlos [Florida State University, Tallahasse, FL (United States). Dept. of Mechanical Engineering and Center for Advanced Power Systems], Emails: martins@caps.fsu.edu, ordonez@eng.fsu.edu

    2010-07-01

    The Alkaline Membrane Fuel Cell (AMFC) is a recently developed fuel cell type, which has shown good experimental results in the laboratory. This paper introduces a mathematical model for the single AMFC with fixed volume and general square section. The main objective is to produce a reliable model (and computationally fast) to predict the response of the single AMFC according to variations of the physical properties of manufacturing materials and operating and design parameters. The model is based on mass, momentum, energy and species conservation, and electrochemical principles, and takes into account pressure drops in the gas channels and temperature gradients with respect to space in the flow direction. The simulation results comprise the AMFC temperature distribution, net power and polarization curves. It is shown that temperature spatial gradients and gas channels pressure drops significantly affect fuel cell performance. Such effects are not usually investigated in the models available in the literature, with most of them assuming uniform pressure and temperature operation. Therefore, the model is expected to be a useful tool for AMFC design and optimization. (author)

  9. Dependence of Regulatory Volume Decrease on Transient Receptor Potential Vanilloid 4 (TRPV4) Expression in Human Corneal Epithelial Cells

    Science.gov (United States)

    Pan, Zan; Yang, Hua; Mergler, Stefan; Liu, Hongshan; Tachado, Souvenir D.; Zhang, Fan; Kao, Winston W. Y.; Koziel, Henry; Pleyer, Uwe; Reinach, Peter S.

    2008-01-01

    TRPV4 is a non-selective cation channel with moderate calcium permeability, which is activated by exposure to hypotonicity. Such a stress induces regulatory volume decrease (RVD) behavior in human corneal epithelial cells (HCEC). We hypothesize that TRPV4 channel mediates RVD in HCEC. Immunohistochemistry revealed centrally and superficially concentrated TRPV4 localization in the corneal tissue. Immunocytochemical and Fluorescence Activated Cell Sorter (FACS) analyses identified TRPV4 membrane surface and cytosolic expression. RT-PCR and Western blot analyses identified TRPV4 gene and protein expression in HCEC, respectively. In addition, 4α-PDD or a 50% hypotonic medium induced up to three-fold transient intracellular Ca2+ ([Ca2+]i) increases. Following TRPV4 siRNA HCEC transfection, its protein expression level declined by 64%, which abrogated these [Ca2+]i transients. Similarly, exposure to either ruthenium red or Ca2+-free Ringer's solution also eliminated this response. In these transfected cells, RVD declined by 51% whereas in the non-transfected counterpart, ruthenium red and Ca2+-free solution inhibited RVD by 54% and 64%, respectively. In contrast, capsazepine, a TRPV1 antagonist, failed to suppress [Ca2+]i transients and RVD. TRPV4 activation contributes to RVD since declines in TRPV4 expression and activity are associated with suppression of this response. In conclusion, there is TRPV4 functional expression in HCEC. PMID:18355916

  10. Sulphonic acid derivatives as probes of pore properties of volume-regulated anion channels in endothelial cells.

    Science.gov (United States)

    Droogmans, G; Maertens, C; Prenen, J; Nilius, B

    1999-09-01

    1. We have used the whole-cell patch-clamp technique to study the effects of 4-sulphonic-calixarenes and some other poly-sulphonic acid agents, such as suramin and basilen blue, on volume-regulated anion channel (VRAC) currents in cultured endothelial cells (CPAE cells). 2. The 4-sulphonic-calixarenes induced a fast inhibition at positive potentials but were ineffective at negative potentials. At small positive potentials, 4-sulphonic-calix[4]arene was a more effective inhibitor than 4-sulphonic-calix[6]arene and -calix[8]arene, which became more effective at more positive potentials. 3. Also suramin and basilen blue induced a voltage dependent current inhibition, reaching a maximum around +40 mV and declining at more positive potentials. 4. The voltage dependence of inhibition was modelled by assuming that these negatively charged molecules bind to a site inside VRAC that senses a fraction delta of the applied electrical field, ranging beween 0.16 to 0.32. 4-Sulphonic-calix[4]arene, suramin and basilen blue bind and occlude VRAC at moderate potentials, but permeate the channel at more positive potentials. 4-Sulphonic-calix[6]arene and -calix[8]arene however do not permeate the channel. From the structural information of the calixarenes, we estimate a lower and upper limit of 11*12 and 17*12 A2 respectively for the cross-sectional area of the pore.

  11. Hypotonic challenge modulates cell volumes differently in the superficial zone of intact articular cartilage and cartilage explant.

    Science.gov (United States)

    Turunen, Siru M; Lammi, Mikko J; Saarakkala, Simo; Koistinen, Arto; Korhonen, Rami K

    2012-05-01

    The objective of this study was to evaluate the effect of sample preparation on the biomechanical behaviour of chondrocytes. We compared the volumetric and dimensional changes of chondrocytes in the superficial zone (SZ) of intact articular cartilage and cartilage explant before and after a hypotonic challenge. Calcein-AM labelled SZ chondrocytes were imaged with confocal laser scanning microscopy through intact cartilage surfaces and through cut surfaces of cartilage explants. In order to clarify the effect of tissue composition on cell volume changes, Fourier Transform Infrared microspectroscopy was used for estimating the proteoglycan and collagen contents of the samples. In the isotonic medium (300 mOsm), there was a significant difference (p integrity of the mechanical environment of chondrocytes.

  12. TRPV4 and AQP4 Channels Synergistically Regulate Cell Volume and Calcium Homeostasis in Retinal Müller Glia

    DEFF Research Database (Denmark)

    Jo, Andrew O; Ryskamp, Daniel A; Phuong, Tam T T

    2015-01-01

    Brain edema formation occurs after dysfunctional control of extracellular volume partly through impaired astrocytic ion and water transport. Here, we show that such processes might involve synergistic cooperation between the glial water channel aquaporin 4 (AQP4) and the transient receptor...... potential isoform 4 (TRPV4), a polymodal swelling-sensitive cation channel. In mouse retinas, TRPV4 colocalized with AQP4 in the end feet and radial processes of Müller astroglia. Genetic ablation of TRPV4 did not affect the distribution of AQP4 and vice versa. However, retinas from Trpv4(-/-) and Aqp4......(-/-) mice exhibited suppressed transcription of genes encoding Trpv4, Aqp4, and the Kir4.1 subunit of inwardly rectifying potassium channels. Swelling and [Ca(2+)]i elevations evoked in Müller cells by hypotonic stimulation were antagonized by the selective TRPV4 antagonist HC-067047 (2-methyl-1...

  13. Comparison of Several Methods for Determining the Internal Resistance of Lithium Ion Cells

    Directory of Open Access Journals (Sweden)

    Hans-Georg Schweiger

    2010-06-01

    Full Text Available The internal resistance is the key parameter for determining power, energy efficiency and lost heat of a lithium ion cell. Precise knowledge of this value is vital for designing battery systems for automotive applications. Internal resistance of a cell was determined by current step methods, AC (alternating current methods, electrochemical impedance spectroscopy and thermal loss methods. The outcomes of these measurements have been compared with each other. If charge or discharge of the cell is limited, current step methods provide the same results as energy loss methods.

  14. Comparison of Several Methods for Determining the Internal Resistance of Lithium Ion Cells

    Science.gov (United States)

    Schweiger, Hans-Georg; Obeidi, Ossama; Komesker, Oliver; Raschke, André; Schiemann, Michael; Zehner, Christian; Gehnen, Markus; Keller, Michael; Birke, Peter

    2010-01-01

    The internal resistance is the key parameter for determining power, energy efficiency and lost heat of a lithium ion cell. Precise knowledge of this value is vital for designing battery systems for automotive applications. Internal resistance of a cell was determined by current step methods, AC (alternating current) methods, electrochemical impedance spectroscopy and thermal loss methods. The outcomes of these measurements have been compared with each other. If charge or discharge of the cell is limited, current step methods provide the same results as energy loss methods. PMID:22219678

  15. Fuel Cell Auxiliary Power Study Volume 1: RASER Task Order 5

    Science.gov (United States)

    Mak, Audie; Meier, John

    2007-01-01

    This study evaluated the feasibility of a hybrid solid oxide fuel cell (SOFC) auxiliary power unit (APU) and the impact in a 90-passenger More-Electric Regional Jet application. The study established realistic hybrid SOFC APU system weight and system efficiencies, and evaluated the impact on the aircraft total weight, fuel burn, and emissions from the main engine and the APU during cruise, landing and take-off (LTO) cycle, and at the gate. Although the SOFC APU may be heavier than the current conventional APU, its weight disadvantage can be offset by fuel savings in the higher SOFC APU system efficiencies against the main engine bleed and extraction during cruise. The higher SOFC APU system efficiency compared to the conventional APU on the ground can also provide considerable fuel saving and emissions reduction, particularly at the gate, but is limited by the fuel cell stack thermal fatigue characteristic.

  16. Cell Shapes and Traction Forces Determine Stress in Motile Confluent Tissue

    Science.gov (United States)

    Yang, Xingbo; Bi, Dapeng; Czajkowski, Michael; Manning, Lisa; Marchetti, Cristina

    Collective cell migration is a highly regulated process involved in wound healing, cancer metastasis and morphogenesis. The understanding of the regulatory mechanism requires the study of mechanical interactions among cells that coordinate their active motion. To this end, we develop a method that determines cellular forces and tissue stresses from experimentally accessible cell shapes and traction forces. This approach allows us for the first time to calculate membrane tensions and hydrostatic pressures at a cellular level in collective migrating cell layers out of equilibrium. It helps us understand the mechanical origin of tissue stresses as previous inferred using Traction Force Microscopy (TFM). We test this approach on a new model of motile confluent tissue, which we term Self-propelled Voronoi Model (SPV) that incorporates cell elasticity, Contractility and motility. With the model, we explore the mechanical properties of confluent motile tissue as a function of cell activities and cell shapes in various geometries.

  17. Cripto regulates skeletal muscle regeneration and modulates satellite cell determination by antagonizing myostatin.

    Science.gov (United States)

    Guardiola, Ombretta; Lafuste, Peggy; Brunelli, Silvia; Iaconis, Salvatore; Touvier, Thierry; Mourikis, Philippos; De Bock, Katrien; Lonardo, Enza; Andolfi, Gennaro; Bouché, Ann; Liguori, Giovanna L; Shen, Michael M; Tajbakhsh, Shahragim; Cossu, Giulio; Carmeliet, Peter; Minchiotti, Gabriella

    2012-11-20

    Skeletal muscle regeneration mainly depends on satellite cells, a population of resident muscle stem cells. However, our understanding of the molecular mechanisms underlying satellite cell activation is still largely undefined. Here, we show that Cripto, a regulator of early embryogenesis, is a novel regulator of muscle regeneration and satellite cell progression toward the myogenic lineage. Conditional inactivation of cripto in adult satellite cells compromises skeletal muscle regeneration, whereas gain of function of Cripto accelerates regeneration, leading to muscle hypertrophy. Moreover, we provide evidence that Cripto modulates myogenic cell determination and promotes proliferation by antagonizing the TGF-β ligand myostatin. Our data provide unique insights into the molecular and cellular basis of Cripto activity in skeletal muscle regeneration and raise previously undescribed implications for stem cell biology and regenerative medicine.

  18. The NF-κB regulator MALT1 determines the encephalitogenic potential of Th17 cells.

    Science.gov (United States)

    Brüstle, Anne; Brenner, Dirk; Knobbe, Christiane B; Lang, Philipp A; Virtanen, Carl; Hershenfield, Brian M; Reardon, Colin; Lacher, Sonja M; Ruland, Jürgen; Ohashi, Pamela S; Mak, Tak W

    2012-12-01

    Effector functions of inflammatory IL-17-producing Th (Th17) cells have been linked to autoimmune diseases such as experimental autoimmune encephalomyelitis (EAE), a mouse model of multiple sclerosis (MS). However, what determines Th17 cell encephalitogenicity is still unresolved. Here, we show that after EAE induction, mice deficient for the NF-κB regulator MALT1 (Malt1-/- mice) exhibit strong lymphocytic infiltration in the CNS, but do not develop any clinical signs of EAE. Loss of Malt1 interfered with expression of the Th17 effector cytokines IL-17 and GM-CSF both in vitro and in vivo. In line with their impaired GM-CSF secretion, Malt1-/- Th cells failed to recruit myeloid cells to the CNS to sustain neuroinflammation, whereas autoreactive WT Th cells successfully induced EAE in Malt1-/- hosts. In contrast, Malt1 deficiency did not affect Th1 cells. Despite their significantly decreased secretion of Th17 effector cytokines, Malt1-/- Th17 cells showed normal expression of lineage-specific transcription factors. Malt1-/- Th cells failed to cleave RelB, a suppressor of canonical NF-κB, and exhibited altered cellular localization of this protein. Our results indicate that MALT1 is a central, cell-intrinsic factor that determines the encephalitogenic potential of inflammatory Th17 cells in vivo.

  19. Visualization of Cell Cycle Variations and Determination of Nucleation in Postnatal Cardiomyocytes.

    Science.gov (United States)

    Raulf, Alexandra; Voeltz, Nadine; Korzus, Daniel; Fleischmann, Bernd K; Hesse, Michael

    2017-02-24

    Cardiomyocytes are prone to variations of the cell cycle, such as endoreduplication (continuing rounds of DNA synthesis without karyokinesis and cytokinesis) and acytokinetic mitosis (karyokinesis but no cytokinesis). Such atypical cell cycle variations result in polyploid and multinucleated cells rather than in cell division. Therefore, to determine cardiac turnover and regeneration, it is of crucial importance to correctly identify cardiomyocyte nuclei, the number of nuclei per cell, and their cell cycle status. This is especially true for the use of nuclear markers for identifying cell cycle activity, such as thymidine analogues Ki-67, PCNA, or pHH3. Here, we present methods for recognizing cardiomyocytes and their nuclearity and for determining their cell cycle activity. We use two published transgenic systems: the Myh6-H2B-mCh transgenic mouse line, for the unequivocal identification of cardiomyocyte nuclei, and the CAG-eGFP-anillin mouse line, for distinguishing cell division from cell cycle variations. Combined together, these two systems ease the study of cardiac regeneration and plasticity.

  20. Near-infrared diffuse reflectance spectroscopy with sample spots and chemometrics for fast determination of bovine serum albumin in micro-volume samples

    Institute of Scientific and Technical Information of China (English)

    Cai-Jing Cui; Wen-Sheng Cai; Xue-Guang Shao

    2013-01-01

    Near-infrared diffuse reflectance spectroscopy (NIRDRS) has attracted more and more attention in analyzing the components in samples with complex matrices.However,to apply this technique to micro-analysis,there are still some obstacles to overcome such as the low sensitivity and spectral overlapping associated with this approach.A method for fast determination of bovine serum albumin (BSA) in micro-volume samples was studied using NIRDRS with sample spots and chemometric techniques.10 μL of sample spotted on a filter paper substrate was used for the spectral measurements.Quantitative analysis was obtained by partial least squares (PLS) regression with signal processing and variable selection.The results show that the correlation coefficient (R) between the predicted and the reference concentration is 0.9897 and the recoveries are in the range of 87.4%-114.4% for the validation samples in the concentration range of 0.61-8.10 mg/mL.These results suggest that the method has the potential to quickly measure proteins in micro-volume solutions.

  1. The gametic central cell of Arabidopsis determines the lifespan of adjacent accessory cells.

    Science.gov (United States)

    Kägi, Christina; Baumann, Nadine; Nielsen, Nicola; Stierhof, York-Dieter; Gross-Hardt, Rita

    2010-12-21

    Plant germ cells develop in specialized haploid structures, termed gametophytes. The female gametophyte patterns of flowering plants are diverse, with often unknown adaptive value. Here we present the Arabidopsis fiona mutant, which forms a female gametophyte that is structurally and functionally reminiscent of a phylogenetic distant female gametophyte. The respective changes include a modified reproductive behavior of one of the female germ cells (central cell) and an extended lifespan of three adjacent accessory cells (antipodals). FIONA encodes the cysteinyl t-RNA synthetase SYCO ARATH (SYCO), which is expressed and required in the central cell but not in the antipodals, suggesting that antip