930 resultados para Flow-cytometric analysis
Resumo:
In [4], Guillard and Viozat propose a finite volume method for the simulation of inviscid steady as well as unsteady flows at low Mach numbers, based on a preconditioning technique. The scheme satisfies the results of a single scale asymptotic analysis in a discrete sense and comprises the advantage that this can be derived by a slight modification of the dissipation term within the numerical flux function. Unfortunately, it can be observed by numerical experiments that the preconditioned approach combined with an explicit time integration scheme turns out to be unstable if the time step Dt does not satisfy the requirement to be O(M2) as the Mach number M tends to zero, whereas the corresponding standard method remains stable up to Dt=O(M), M to 0, which results from the well-known CFL-condition. We present a comprehensive mathematical substantiation of this numerical phenomenon by means of a von Neumann stability analysis, which reveals that in contrast to the standard approach, the dissipation matrix of the preconditioned numerical flux function possesses an eigenvalue growing like M-2 as M tends to zero, thus causing the diminishment of the stability region of the explicit scheme. Thereby, we present statements for both the standard preconditioner used by Guillard and Viozat [4] and the more general one due to Turkel [21]. The theoretical results are after wards confirmed by numerical experiments.
Resumo:
Para el presente trabajo realizado en la Universidad del Rosario, buscamos hacer un mejoramiento productivo de la empresa Apparel Basic Ltda. Teniendo en cuenta todas las herramientas aprehendidas durante la academia y aplicando esto a una empresa del sector real de confecciones. Se centrara el análisis principalmente en las tres áreas donde se observan los mayores problemas organizacionales: Producción, manejo de inventarios y entrega de producto. En el primer análisis se realizara todo el estudio del proceso de producción, teniendo en cuenta la distribución en planta, las rutas críticas de proceso, los diagramas de flujo de producto, el análisis de las principales referencias, entre otros. Todo esto con el fin de identificar los principales errores y poder proponer herramientas y procesos de mejora que sean de ayuda para esta organización. El siguiente análisis se desarrollara en el manejo de inventarios; dentro de este aspecto se analizaran la distribución de las bodegas de producto, la identificación de los productos de mayor rotación, el planteamiento de indicadores de gestión, entre otros procesos, con el fin de identificar procesos benéficos para la empresa que aceleren y mejoren el flujo de producto al interior de la organización. Luego de esto, se analiza todo el proceso de alistamiento y entrega de producto ya que es uno de los principales problemas dentro de la organización porque se esta incumpliendo con los pedidos de los clientes lo que genera un problema de insatisfacción por parte de los clientes.
Resumo:
Sodium chloride-induced cell and nuclear degradation in the root meristems of sweetpotato [Ipomoea batatas (L.) Lam.] were determined using fluorescent microscopy and flow cytometry analysis. Two sweetpotato cultivars were grown in liquid Murashige and Skoog medium and subjected to 0 mM and 500 mM NaCl, with or without 15 mM CaCl2, for periods up to 24 h. Changes to the nuclei of root meristematic cells showed a similar pattern of damage to the nuclei using both fluorescent microscopy and flow cytometry analysis. Damage occurring after only a few hours was followed by nuclear degradation at 24 h. Flow cytometry histograms showed a reduction in G1 and G2 nuclei and an increase in degraded nuclei in NaCl-stressed roots. Salinity-induced nuclear degradation was alleviated by the addition of CaCl2.
Resumo:
Abnormal vascular smooth muscle cell (VSMC) proliferation plays an important role in the pathogenesis of both atherosclerosis and restenosis. Recent studies suggest that high-dose salicylates, in addition to inhibiting cyclooxygenase activity, exert an antiproliferative effect on VSMC growth both in-vitro and in-vivo. However, whether all non-steroidal anti-inflammatory drugs (NSAIDs) exert similar anti proliferative effects on VSMCs, and do so via a common mechanism of action, remains to be shown. In this study, we demonstrate that the NSAIDs aspirin, sodium salicylate, diclofenac, ibuprofen, indometacin and sulindac induce a dose-dependent inhibition of proliferation in rat A10 VSMCs in the absence of significant cytotoxicity. Flow cytometric analyses showed that exposure of A10 cells to diclofenac, indometacin, ibuprofen and sulindac, in the presence of the mitotic inhibitor, nocodazole, led to a significant G0/G1 arrest. In contrast, the salicylates failed to induce a significant G1 arrest since flow cytometry profiles were not significantly different from control cells. Cyclin A levels were elevated, and hyperphosphorylated p107 was present at significant levels, in salicylate-treated A10 cells, consistent with a post-G1/S block, whereas cyclin A levels were low, and hypophosphorylated p107 was the dominant form, in cells treated with other NSAIDs consistent with a G1 arrest. The ubiquitously expressed cyclin-dependent kinase (CDK) inhibitors, p21 and p27, were increased in all NSAID-treated cells. Our results suggest that diclofenac, indometacin, ibuprofen and sulindac inhibit VSMC proliferation by arresting the cell cycle in the G1 phase, whereas the growth inhibitory effect of salicylates probably affects the late S and/or G2/M phases. Irrespective of mechanism, our results suggest that NSAIDs might be of benefit in the treatment of certain vasculoproliferative disorders.
Resumo:
Abnormal vascular smooth muscle cell (VSMC) proliferation is known to play an important role in the pathogenesis of atherosclerosis, restenosis and instent stenosis. Recent studies suggest that salicylates, in addition to inhibiting cyclooxygenase activity, exert an antiproliferative effect on VSMC growth both in vitro and in vivo. However, whether all non-steroidal anti-inflammatory drugs (NSAID) exert similar antiproliferative effects on VSMCs, and do so via a common mechanism of action, remains unknown. In the present study, we demonstrated that the NSAIDs, aspirin, ibuprofen and sulindac induced a dose-dependent inhibition of proliferation in rat A10 VSMCs (IC50 = 1666 mumol/L, 937 mumol/L and 520 mumol/L, respectively). These drugs did not show significant cytotoxic effects as determined by LDH release assay, even at the highest concentrations tested (aspirin, 5000 mumol/L; ibuprofen, 2500 mumol/L; and sulindac, 1000 mumol/L). Flow cytometric analyses showed that a 48 h exposure of A10 VSMCs to ibuprofen (1000 mumol/L) and sulindac (750 mumol/L) led to a significant G1 arrest (from 68.7 +/- 2.0% of cells in G1 to 76.6 +/- 2.2% and 75.8 +/- 2.2%, respectively, p < 0.05). In contrast, aspirin (2500 mumol/L) failed to induce a significant G1 arrest (68.1 +/- 5.2%). Clearer evidence of a G1 block was obtained by treatment of cells with the mitotic inhibitor, nocodazole (40 ng/ml), for the final 24 h of the experiment. Under these conditions, aspirin still failed to induce a G1 arrest (from 25.9 +/- 10.9% of cells in G1 to 19.6 +/- 2.3%) whereas ibuprofen and sulindac led to a significant accumulation of cells in G1(51.8% +/- 17.2% and 54.1% +/- 10.6%, respectively, p < 0.05). These results indicate that ibuprofen and sulindac inhibit VSMC proliferation by arresting the cell cycle in the G1 phase whereas the effect of aspirin appears to be independent of any special phase of the cell cycle. Irrespective of mechanism, our results suggest that NSAIDs might be of benefit to the treatment of vascular proliferative disorders.
Resumo:
The genome structure of Colletotrichum lindemuthianum in a set of diverse isolates was investigated using a combination of physical and molecular approaches. Flow cytometric measurement of genome size revealed significant variation between strains, with the smallest genome representing 59% of the largest. Southern-blot profiles of a cloned fungal telomere revealed a total chromosome number varying from 9 to 12. Chromosome separations using pulsed-field gel electrophoresis (PFGE) showed that these chromosomes belong to two distinct size classes: a variable number of small (< 2.5 Mb) polymorphic chromosomes and a set of unresolved chromosomes larger than 7 Mb. Two dispersed repeat elements were shown to cluster on distinct polymorphic minichromosomes. Single-copy flanking sequences from these repeat-containing clones specifically marked distinct small chromosomes. These markers were absent in some strains, indicating that part of the observed variability in genome organization may be explained by the presence or absence, in a given strain, of dispensable genomic regions and/or chromosomes.
Resumo:
BACKGROUND: Evidence suggests the wide variation in platelet response within the population is genetically controlled. Unraveling the complex relationship between sequence variation and platelet phenotype requires accurate and reproducible measurement of platelet response. OBJECTIVE: To develop a methodology suitable for measuring signaling pathway-specific platelet phenotype, to use this to measure platelet response in a large cohort, and to demonstrate the effect size of sequence variation in a relevant model gene. METHODS: Three established platelet assays were evaluated: mobilization of [Ca(2+)](i), aggregometry and flow cytometry, each in response to adenosine 5'-diphosphate (ADP) or the glycoprotein (GP) VI-specific crosslinked collagen-related peptide (CRP). Flow cytometric measurement of fibrinogen binding and P-selectin expression in response to a single, intermediate dose of each agonist gave the best combination of reproducibility and inter-individual variability and was used to measure the platelet response in 506 healthy volunteers. Pathway specificity was ensured by blocking the main subsidiary signaling pathways. RESULTS: Individuals were identified who were hypo- or hyper-responders for both pathways, or who had differential responses to the two agonists, or between outcomes. 89 individuals, retested three months later using the same methodology, showed high concordance between the two visits in all four assays (r(2) = 0.872, 0.868, 0.766 and 0.549); all subjects retaining their phenotype at recall. The effect of sequence variation at the GP6 locus accounted for approximately 35% of the variation in the CRP-XL response. CONCLUSION: Genotyping-phenotype association studies in a well-characterized, large cohort provides a powerful strategy to measure the effect of sequence variation in genes regulating the platelet response.
Resumo:
Observations by the EISCAT experiments “POLAR” and Common Programme CP-3 reveal non-Maxwellian ion velocity distributions in the auroral F-region ionosphere. Analysis of data from three periods is presented. During the first period, convection velocities are large (≈2 km s-1) and constant over part of a CP-3 latitude scan; the second period is one of POLAR data containing a short-lived (<1 min.) burst of rapid (>1.5 km s-1) flow. We concentrate on these two periods as they allow the study of a great many features of the ion-neutral interactions which drive the plasma non-thermal and provide the best available experimental test for models of the 3-dimensional ion velocity distribution function. The third period is included to illustrate the fact that non-thermal plasma frequently exists in the auroral ionosphere: the data, also from the POLAR experiment, cover a three-hour period of typical auroral zone flow and analysis reveals that the ion distribution varies from Maxwellian to the threshold of a toroidal form.
Resumo:
Background: The role of osteocytes in bone structure and function remains partially unresolved. Their participation in mechanotransduction, i.e., the conversion of a physical stimulus into a cellular response, has been hypothesized. The present study was an evaluation of the osteocyte density in the peri-implant bone of immediately loaded and submerged dental implants. Methods: Fourteen male patients were included in the study; all of them were partially edentulous and needed a posterior mandibular restoration. Implants were inserted in these areas; half of the sample was loaded immediately (included in a fixed provisional prosthesis on the same day as implant surgery), whereas the other half was left to heal submerged. Fourteen implants (seven immediately loaded and seven unloaded) were retrieved with a trephine after a healing period of 8 weeks. The specimens were treated to obtain thin ground sections, and histomorphometry was used to evaluate the osteocyte index in the peri-implant bone. Results: A higher and statistically significant number of osteocytes was found in the peri-implant bone around immediately loaded implants (P=0.0081). A correlation between the percentage of bone-implant contact and osteocyte density was found for immediately loaded implants (P=0.0480) but not for submerged implants (P=0.2667). Conclusion: The higher number of osteocytes in the peri-implant bone around immediately loaded implants could be related to the functional adaptation required by the loading stimulus, which also explains the hypothesized involvement of the osteocytes in the maintenance of the bone matrix. J Periodontol 2009;80:499-504.
Resumo:
Large pore ordered mesoporous silica FDU-1 with three-dimensional (3D) face-centered cubic, Fm3m arrangement of rnesopores, was synthesized under strong acid media using B-50-6600 poly(ethylene oxide)-poly(butylene oxide)-poly(ethylene oxide) triblock copolymer (EO(39)BO(47)EO(39)), tetraethyl orthosilicate (TEOS) and trimethyl-benzene (TMB). Large pore FDU-1 silica was obtained by using the following gel composition 1TEOS:0.00735B50-6600:0.00735TMB:6HCl:155H(2)O. The pristine material exhibited a BET specific surface area of 684 m(2) g(-1), total pore volume of 0.89 cm(3) g(-1), external surface area of 49 m(2) g(-1) and microporous volume of 0.09 cm(3) g(-1). The enzyme activity was determined by the Flow Injection Analysis-Chemiluminescence (FIA-CL) method. For GOD immobilized on the FDU-1 silica, GOD supernatant and GOD solution, the FIA-CL results were 9.0, 18.6 and 34.0 U, respectively. The value obtained for the activity of the GOD solution with FIA-CL method is in agreement with the 35 U, obtained by spectrophotometry. (C) 2011 Elsevier B.V. All rights reserved.
Resumo:
We describe the development of a label free method to analyze the interactions between Ca(2+) and the porcine S100A12 protein immobilized on polyvinyl butyral (PVB). The modified gold electrodes were characterized using cyclic voltammetry (CV), electrochemical impedance spectroscopy (EIS), scanning electron microscopy (SEM) and surface plasmon resonance (SPR) techniques. SEM analyses of PVB and PVB-S100A12 showed a heterogeneous distribution of PVB spherules on gold surface. EIS and CV measurements have shown that redox probe reactions on the modified gold electrodes were partially blocked due the adsorption of PVB-S100A12, and confirm the existence of a positive response of the immobilized S100Al2 to the presence of calcium ions. The biosensor exhibited a wide linear response to Ca(2+) concentrations ranging from 12.5 to 200 mM. The PVB-S100A12 seems to be bound to the gold electrode surface by physical adsorption: we observed an increase of 1184.32 m degrees in the SPR angle after the adsorption of the protein on the PVB surface (in an indication that 9.84 ng of S100A12 are adsorbed per mm(2) of the Au-PVB electrode), followed by a further increase of 581.66 m degrees after attachment of the Ca(2+) ions. In addition, no SPR response is obtained for non-specific ions. These studies might be useful as a platform for the design of new reusable and sensitive biosensing devices that could find use in the clinical applications. (C) 2010 Elsevier B.V. All rights reserved.
Resumo:
Magnetic fabric and rock magnetism studies were performed on 25 unmetamorphosed mafic dikes of the Meso-Late Proterozoic (similar to 1.02 Ga) dike swarm from Salvador (Bahia State, NE Brazil). This area lies in the north-eastern part of the Sao Francisco Craton, which was dominantly formed/reworked during the Transamazonian orogeny (2.14-1.94 Ga). The dikes crop out along the beaches and in quarries around Salvador city, and cut across both amphibolite dikes and granulites. Their widths range from a few centimeters up to 30 m with an average of similar to 4 m, and show two main trends N 140-190 and N 100-120 with vertical dips. Magnetic fabrics were determined using both anisotropy of low-field magnetic susceptibility (AMS) and anisotropy of anhysteretic remanent magnetization (AARM). The magnetic mineralogy was investigated by many experiments including remanent magnetization measurements at variable low temperatures (10-300 K), Mossbauer spectroscopy, high temperature magnetization curves (25-700 degrees C) and scanning electron microscopy (SEM). The rock magnetism study suggests pseudo-single-domain magnetite grains carrying the bulk magnetic susceptibility and AARM fabrics. The magnetite grains found in these dikes are large and we discard the presence of single-domain grains. Its composition is close to stoichiometric with low Ti substitution, and its Verwey transition occurs around 120 K. The main AMS fabric recognized in the swarm is so-called normal, in which the K(max)-K(int) plane is parallel to the dike plane and the magnetic foliation pole K(min)) is perpendicular to it. This fabric is interpreted as due to magma flow, and analysis of the K m inclination permitted to infer that approximately 80% of the dikes were fed by horizontal or sub-horizontal flows (K(max) < 30 degrees). This interpretation is supported by structural field evidence found in five dikes. In addition, based on the plunge of K(max), two mantle sources could be inferred; one of them which fed about 80% of the swarm would be located in the southern part of the region, and the other underlied the Valeria quarry. However, for all dikes the AARM tensors are not coaxial with AMS fabrics and show a magnetic lineation (AARM(max)) oriented to N30-60E, suggesting that magnetite grains were rotated clockwise from dike plane. The orientation of AARM lineation is similar to the orientation of a system of faults in which the Salvador normal fault is the most important. These faults were formed during Cretaceous rifting in the Reconcavo-Tucano-jatoba assemblage that corresponds to an aborted intra-continental rift formed during the opening of the South Atlantic. Therefore, the AARM fabric found for the Salvador dikes is probably tectonic in origin and suggests that the dike swarm was affected by the important tectonic event responsible for the break-up of the Gondwanaland. (C) 2008 Elsevier B.V. All rights reserved.
Resumo:
This paper describes the applications of anew carbon paste electrode containing fibers of coconut (Cocus nucifera L) fruit, which are very rich in peroxidase enzymes naturally immobilized on its structure. The new sensor was applied for the amperometric quantification of benzoyl peroxide in facial creams and dermatological shampoos. The amperometric measurements were performed in 0.1 mol L(-1) phosphate buffer (pH 5.2), at 0.0 V (versus Ag/AgCl). On these conditions, benzoyl peroxide was rapidly determined in the 5.0-55 mu mol L(-1), with a detection limit of 2.5 mu mol L(-1) (s/n = 3), response time of 4.1 s (90% of the steady state) and sensitivity limit of 0.33 A mol L(-1) cm(-2). The amperometric results are in good agreement with those obtained by spectrophotometric technique, used as a standard method. (C) 2009 Elsevier B.V. All rights reserved.
Resumo:
A sensor for H2O2 amperometric detection based on a Prussian blue (PB) analogue was developed. The electrocatalytic process allows the determination of hydrogen peroxide at 0.0 V with a limit of detection of 1.3 mu mol L-1 in a flow injection analysis (FIA) configuration. Studies on the optimization of the FIA parameters were performed and under optimal FIA operational conditions the linear response of the method was extended up to 500 mu mol L-1 hydrogen peroxide with good stability. The possibility of using the developed sensor in medium containing sodium ions and the increased operational stability constitute advantages in comparison with PB-based amperometric sensors. The usefulness of the methodology was demonstrated by addition-recovery experiments with rainwater samples and values were in the 98.8 to 103% range.
Resumo:
The anodic oxidation of ascorbic acid on a ruthenium oxide hexacyanoferrate modified electrode was characterized by cyclic voltammetry. On this modified surface, the electrocatalytic process allows the determination of ascorbic acid to be performed at 0.0 V and pH 6.9 with a limit of detection of 2.2 mu M in a flow injection configuration. Under this experimental condition, no interference from glucose, nitrite and uric acid was noticed. Lower detection limit values were obtained by measuring flow injection analysis (FIA) responses at 0.4 V (0.14 mu M), but a concurrent loss of selectivity is expected at this more positive potential. Under optimal FIA operating conditions, the linear response of the method was extended up to 1 mM ascorbic acid. The repeatability of the method for injections of a 1.0 mM ascorbic acid solution was 2.0% (n=10). The usefulness of the method was demonstrated by an addition-recovery experiment with urine samples and the recovered values were in the 98-104% range. (c) 2007 Elsevier B.V. All rights reserved.