Sample records for fluid cell count

  1. Body fluid cell counts by automated methods.

    PubMed

    Sandhaus, Linda M

    2015-03-01

    Automated cell counting for body fluids is gradually replacing manual cell counting by hemocytometer. Automation offers potential benefits of improved accuracy, efficiency, and standardization. The addition of body fluid modes to some hematology analyzers adapts the technology and software to meet the particular requirements of body fluid analysis. However, the functional sensitivity for low cell counts currently limits applicability of automated methods to all types of body fluid. Microscopic review is indicated when malignancy is a diagnostic consideration. PMID:25676374

  2. Automated counting of cells in cerebrospinal fluid using the CellDyn-4000 haematology analyser.

    PubMed

    Hoffmann, Johannes J M L; Janssen, Willy C M

    2002-11-01

    Counting of cells in cerebrospinal fluid is currently performed manually. Because of the inherent analytical and economical disadvantages, we attempted to introduce a fully automated method. Therefore, we validated the Abbott CellDyn-4000 haematology analyser for counting cells in cerebrospinal fluid. The analyser was used in its standard configuration with the simple precaution of a preceding blank sample. As for leukocyte counting the analyser yielded high precision (CV approximately 5% above the upper reference limit), good linearity, low limit of detection (2/microl) and excellent correlation (r > 0.99) with the counting chamber method. The differential leukocyte count was equally accurate and precise, even in the low concentration range. Performance of the erythrocyte count was impaired by its high limit of detection (6/nl) and it appeared satisfactory only for detecting blood admixture due to traumatic puncture. The specificity of the analyser is excellent, since it correctly classified non-viable leukocytes and excluded yeast cells from the leukocyte count in a patient with cryptococcal meningitis. We conclude that the CellDyn-4000 is well suited for quickly and reliably counting leukocytes in cerebrospinal fluid. Developing some software modifications might make the analyser useful also for performing erythrocyte counting in cerebrospinal fluid. PMID:12521237

  3. Validation of automated blood cell counter for the determination of polymorphonuclear cell count in the ascitic fluid of cirrhotic patients with or without spontaneous bacterial peritonitis

    Microsoft Academic Search

    Stefania Angeloni; Giorgia Nicolini; Manuela Merli; Francesca Nicolao; Giorgio Pinto; Teresa Aronne; Adolfo Francesco Attili; Oliviero Riggio

    2003-01-01

    ObjectivePolymorphonuclear (PMN) cell count in ascitic fluid is the most useful test for the diagnosis of spontaneous bacterial peritonitis (SBP). We evaluated the validity of an automated blood cell counter for the PMN determination in ascitic fluid by comparing it with the traditional hematologic method with a light microscope in a manual counting chamber.

  4. Validation of automated blood cell counter for the determination of polymorphonuclear cell count in the ascitic fluid of cirrhotic patients with or without spontaneous bacterial peritonitis

    Microsoft Academic Search

    Stefania Angeloni; Giorgia Nicolini; Manuela Merli; Francesca Nicolao; Giorgio Pinto; Teresa Aronne; Adolfo Francesco Attili; Oliviero Riggio

    2003-01-01

    OBJECTIVE:Polymorphonuclear (PMN) cell count in ascitic fluid is the most useful test for the diagnosis of spontaneous bacterial peritonitis (SBP). We evaluated the validity of an automated blood cell counter for the PMN determination in ascitic fluid by comparing it with the traditional hematologic method with a light microscope in a manual counting chamber.METHODS:A total of 130 ascitic fluid samples

  5. White blood cell counting system

    NASA Technical Reports Server (NTRS)

    1972-01-01

    The design, fabrication, and tests of a prototype white blood cell counting system for use in the Skylab IMSS are presented. The counting system consists of a sample collection subsystem, sample dilution and fluid containment subsystem, and a cell counter. Preliminary test results show the sample collection and the dilution subsystems are functional and fulfill design goals. Results for the fluid containment subsystem show the handling bags cause counting errors due to: (1) adsorption of cells to the walls of the container, and (2) inadequate cleaning of the plastic bag material before fabrication. It was recommended that another bag material be selected.

  6. Effects of Somatic Cell Count on Quality and ShelfLife of Pasteurized Fluid Milk1

    Microsoft Academic Search

    Y. Ma; C. Ryan; D. M. Barbano; D. M. Galton; M. A. Rudan; K. J. Boor

    2000-01-01

    Milk was collected from eight Holstein cows four times before and four times after intramammary infec- tion with Streptococcus agalactiae. Postinfection milk had significantly higher somatic cell count (SCC) (849,000 cells\\/ml) than preinfection milk (45,000 cells\\/ ml). High SCC raw milk had more lipolysis and proteol- ysis than low SCC raw milk. Pasteurized, homogenized, 2% fat milks from pre- and

  7. A method for counting monosodium urate crystals in synovial fluid.

    PubMed

    Montagna, P; Brizzolara, R; Ferrone, C; Cutolo, M; Paolino, S; Cimmino, M A

    2015-01-01

    This study was aimed to standardize the technique for counting monosodium urate (MSU) crystals in the synovial fluid (SF) of patients with gout. A total of 52 SF specimens were examined under a polarized light microscope. The amount of SF ranged between 0.1 and 45 mL (median 3 mL). MSU crystals were counted in four areas with the same size at 400x magnification. Cytological examination of the same specimens was also performed. Median leukocyte count was 400 cells/mm3 (range 50-14,000 cells/mm3), with a median percentage of polymorphonuclear leukocytes of 9% (range 0%-98%). Median crystal count was 179.5 (range 3-1600). Inter- reader and intra-reader agreement in crystal counting were good with a weighed k of 0.89 [95% confidence interval (CI) 0.85-0.94] and 0.89 (95% CI 0.84-0.93), respectively. Our data indicate that the SF MSU crystal count is a feasible and highly reliable technique. PMID:26150273

  8. Somatic Cell Counts in Holsteins

    Microsoft Academic Search

    B. W. KENNEDY; M. S. SETHAR; A. K. W. TONG; J. E. MOXLEY; B. R. DOWNEY; Dairy Herd

    Between February and December, 1977, 133,493 test-day observations of somatic cell count were taken on 27,009 Holstein cows in 676 herds on the Quebec Dairy Herd Analysis Service. Data were transformed to a log (natural) scale, and analyses were separate within lacta- tion age group (42, 3, 4, 5, and \\/>6 yr). Joint estimates of fixed effects of month of

  9. White blood cell counts: reference methodology.

    PubMed

    Chabot-Richards, Devon S; George, Tracy I

    2015-03-01

    Modern hematology laboratories use automated hematology analyzers to perform cell counts. These instruments provide accurate, precise, low-cost differential counts with fast turnaround times. Technologies commonly used include electrical impedance, radiofrequency conductivity, laser light scattering, and cytochemistry. This article reviews the principles of these methodologies and possible sources of error, provides guidance for selecting flagging criteria, and discusses novel, clinically relevant white blood cell parameters provided by new instruments, including immature granulocyte count and granularity index. PMID:25676369

  10. Automatic counting of immunocytochemically stained cells

    Microsoft Academic Search

    F. Arambula Cosio; J. A. Marquez Flores; M. A. Padilla Castaneda; S. Solano; P. Tato

    2003-01-01

    In this work is described the development of an automatic color image segmentation and cell counting system for immunocytochemical analysis of stained tissue samples. The system is designed to automatically count the total number of positive and negative cells in tissue samples treated with cytokines DNA probes of pigs naturally parasitized with Taenia solium metacestodes and using in situ hybridization.

  11. Obesity and Immune Cell Counts in Women

    PubMed Central

    Womack, Julie; Tien, Phyllis C.; Feldman, Joseph; Shin, Ja Hyun; Fennie, Kristopher; Anastos, Kathryn; Cohen, Mardge H.; Bacon, Melanie C.; Minkoff, Howard

    2007-01-01

    Objective Obesity is common in women and associated with a number of adverse health outcomes including cardiovascular disease, infectious diseases, and cancer. We explore the relationship between obesity and immune cell counts in women. Design Longitudinal study of 322 women from 1999 through 2003 enrolled as HIV-negative comparators in the Women’s Interagency HIV Study. Methods Body mass index (BMI) was categorized as normal weight (BMI 18.5 - 24.9), overweight (BMI 25 - 29.9), obese (BMI 30 - 34.9), and morbid obesity (BMI ?35). CD4 and CD8 counts and percents, total lymphocyte and white blood cell (WBC) counts were measured annually using standardized techniques. A mixed model repeated measures analysis was performed using an autoregressive correlation matrix. Results At the index visit, 61% of women were African-American; mean age was 35 years, and median BMI was 29 kg/m2. Immunologic parameters were in the normal range (median CD4 count: 995 cells/mm3; CD8 count: 488 cells/mm3; total lymphocyte count: 206 cells/mm3; median WBC: 6 × 103 cells/mm3). In multivariate analyses, being overweight, obese or morbidly obese were independently associated with higher CD4, total lymphocyte, and WBC counts than being normal weight; morbid obesity was associated with a higher CD8 count. The strongest associations between body weight and immune cell counts were demonstrated in the morbidly obese. Conclusion Increasing body weight is associated with higher CD4, CD8, total lymphocyte, and WBC counts in women. Investigation into the impact of obesity on immune function and long term adverse outcomes is needed. PMID:17570264

  12. Trapping cells in paper for white blood cell count.

    PubMed

    Zhang, Yi; Bai, Jianhao; Wu, Hong; Ying, Jackie Y

    2015-07-15

    White blood cell count is an important indicator of each individual's health condition. An abnormal white blood cell count usually results from an infection, cancer, or other conditions that trigger systemic inflammation responses. White blood cell count also provides predictive information on the incidence of cardiovascular diseases and Type 2 diabetes. Therefore, monitoring white blood cell count on a regular basis can potentially help individuals to take preventive measures and improve healthcare outcomes. Currently, white blood cell count is primarily conducted in centralized laboratories, and it requires specialized equipment and dedicated personnel to perform the test and interpret the results. So far there has been no rapid test that allows white blood cell count in low-resource settings. In this study, we have demonstrated a vertical flow platform that quantifies white blood cells by trapping them in the paper. White blood cells were tagged with gold nanoparticles, and flowed through the paper via a small orifice. The white blood cell count was determined by measuring the colorimetric intensity of gold nanoparticles on the surface of white blood cells that were trapped in the paper mesh. Using this platform, we were able to quantify white blood cells in 15?L of blood, and visually differentiate the abnormal count of white blood cells from the normal count. The proposed platform enabled rapid white blood cell count in low resource settings with a small sample volume requirement. Its low-cost, instrument-free operations would be attractive for point-of-care applications. PMID:25721975

  13. CELL TYPES, DIFFERENTIAL CELL COUNTS, AND BLOOD CELL MEASUREMENTS OF

    E-print Network

    count than those repolted for other sharks. 10 Microns I I FIGURE I.-Mitosis-Prophase. Cell Differentials FIGURE 2.-Mitosis-Anaphase. The mean size of the POltuguese shark mature erythrocytes (Figure 3

  14. Why Count Types of White Blood Cells?

    NSDL National Science Digital Library

    Ethel D. Stanley (Beloit College; Biology)

    2006-05-20

    How can we make use of complex cellular level responses in the human body to microbial infections and other disorders? Why is it important to differentiate between white blood cells in a blood sample and keep a record of their numbers? Improve skills at cell identification and explore these questions with the program Cell Differentials. * identify lymphocytes in a clinical laboratory simulation of blood cell counts

  15. Monitoring udder health and milk quality using somatic cell counts

    Microsoft Academic Search

    Ynte H. Schukken; David J. Wilson; Francis Welcome; Linda Garrison-Tikofsky; Ruben N. Gonzalez

    2003-01-01

    In this article the use of somatic cell counts for monitoring udder health and milk quality is discussed. Somatic cell count dynamics at quarter, cow, herd and population level are discussed and illustrated with examples. Quarter and cow somatic cell counts directly represent the inflammatory status of the mammary gland. Herd and population somatic cell count are related to the

  16. Differential white cell count by centrifugal microfluidics.

    SciTech Connect

    Sommer, Gregory Jon; Tentori, Augusto M.; Schaff, Ulrich Y.

    2010-07-01

    We present a method for counting white blood cells that is uniquely compatible with centrifugation based microfluidics. Blood is deposited on top of one or more layers of density media within a microfluidic disk. Spinning the disk causes the cell populations within whole blood to settle through the media, reaching an equilibrium based on the density of each cell type. Separation and fluorescence measurement of cell types stained with a DNA dye is demonstrated using this technique. The integrated signal from bands of fluorescent microspheres is shown to be proportional to their initial concentration in suspension. Among the current generation of medical diagnostics are devices based on the principle of centrifuging a CD sized disk functionalized with microfluidics. These portable 'lab on a disk' devices are capable of conducting multiple assays directly from a blood sample, embodied by platforms developed by Gyros, Samsung, and Abaxis. [1,2] However, no centrifugal platform to date includes a differential white blood cell count, which is an important metric complimentary to diagnostic assays. Measuring the differential white blood cell count (the relative fraction of granulocytes, lymphocytes, and monocytes) is a standard medical diagnostic technique useful for identifying sepsis, leukemia, AIDS, radiation exposure, and a host of other conditions that affect the immune system. Several methods exist for measuring the relative white blood cell count including flow cytometry, electrical impedance, and visual identification from a stained drop of blood under a microscope. However, none of these methods is easily incorporated into a centrifugal microfluidic diagnostic platform.

  17. 21 CFR 864.8185 - Calibrator for red cell and white cell counting.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ...2014-04-01 false Calibrator for red cell and white cell counting. 864...Reagents § 864.8185 Calibrator for red cell and white cell counting. (a) Identification. A calibrator for red cell and white cell counting is a...

  18. 21 CFR 864.8185 - Calibrator for red cell and white cell counting.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ...2011-04-01 false Calibrator for red cell and white cell counting. 864...Reagents § 864.8185 Calibrator for red cell and white cell counting. (a) Identification. A calibrator for red cell and white cell counting is a...

  19. 21 CFR 864.8185 - Calibrator for red cell and white cell counting.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ...2012-04-01 false Calibrator for red cell and white cell counting. 864...Reagents § 864.8185 Calibrator for red cell and white cell counting. (a) Identification. A calibrator for red cell and white cell counting is a...

  20. 21 CFR 864.8185 - Calibrator for red cell and white cell counting.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ...2013-04-01 false Calibrator for red cell and white cell counting. 864...Reagents § 864.8185 Calibrator for red cell and white cell counting. (a) Identification. A calibrator for red cell and white cell counting is a...

  1. 21 CFR 864.8185 - Calibrator for red cell and white cell counting.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ...2010-04-01 false Calibrator for red cell and white cell counting. 864...Reagents § 864.8185 Calibrator for red cell and white cell counting. (a) Identification. A calibrator for red cell and white cell counting is a...

  2. Bacterial Cell Counts in Goat Milk and Their Correlations with Somatic Cell Counts, Percent Fat, and Protein1

    Microsoft Academic Search

    Y. W. Park; R. D. Humphrey

    1986-01-01

    Representative milk samples at morning and afternoon milking were collected periodically for 5 mo from 32 does in a Prairie View A&M University milking herd to test the concentrations of total bacterial, coliform, and staphylococcus counts and to determine the correlations among the bacterial cell counts, somatic cell counts, percent fat, and percent protein. Bacterial cell counts were assayed by

  3. Cytospin centrifuge in differential counts of milk somatic cells.

    PubMed

    Dulin, A M; Paape, M J; Weinland, B T

    1982-07-01

    A procedure was developed for a cytospin centrifuge to concentrate cells from milk onto microscope slides in a circle with 6 mm diameter. Differential somatic cell counts with cytospin were compared to the conventional hand smearing technique to ascertain variation in differential counts. Milk samples were from each of 30 cow quarters, 10 within each of three total milk somatic cell count ranges of less than .7, .7 to 1.5, and more than 1.5 x 10(6)/ml. Two prepared from each sample. All smears were air dried; and stained with modified Wright's stain, the first 200 cells were counted. Differential cell counts from smears prepared by the two procedures were not different. Variation between duplicate smears in differential cell counts was less for cytospin technique. Cytospin can be used to obtain rapid and accurate differential cell counts in milk over a wide range of total somatic cell counts. PMID:7050194

  4. Rapid count of microbial cells in dialysate.

    PubMed

    Shimakita, Tomonori; Yamamoto, Hidenori; Naramura, Tomotaka; Fujimori, Akira; Ide, Takao; Tashiro, Yoshikazu; Saito, Mikako; Matsuoka, Hideaki

    2007-10-01

    An apparatus for the non-culture method (NCM) of microbial cell count was formerly developed and named a bioplorer. The bioplorer NCM is based on the double staining of cells with 4', 6-diamidino-2-phenylindole (DAPI) and propidium iodide (PI) and the automatic analysis of their fluorescent microscopic images. Viable cells can be stained with DAPI, while dead cells can be stained with DAPI and PI. In this study, the bioplorer NCM has been applied to the dialysate. The viable and dead cells in dialysate could be counted within 20 min. The detection limit expressed by log(10)[cells/100 mL] was 2.0. When cell-spiked dialysate samples containing prescribed number of Bacillus subtilis cells were assayed, the numbers of cells determined by the bioplorer NCM (N(VIA)(NCM)) and a conventional culture method (CM) on R2A medium (N(VIA)(R2A-CM)) were similar in the range of 2.6-4.6 within the 95% confidence interval (NCM-CM equivalent range). When test solutions sampled from a practical facility in a hospital were assayed, N(VIA)(NCM) was greater than, but comparable to, N(VIA)(R2A-CM). The endotoxin (ET) in the test samples were assayed as well using a test kit for limulus amoebocyte lysate assay. The results of microbial cells and ET concentration indicated that the dialysate supplying line was clean and well maintained. The bioplorer NCM can determine if the microbial contamination of dialysate supplying facilities is greater than 2.6 (398 cells/100 mL). PMID:17845395

  5. RAPID GLUTARALDEHYDE FIXATION FOR COW MILK CELL COUNT

    E-print Network

    Paris-Sud XI, Université de

    ) is regarded as the reference method for counting somatic cells in cow milk. The use of an electronic particle by the for- maldehyde technique. Materials and Methods Somatic cell counts in 40 samples of milk were carriedRAPID GLUTARALDEHYDE FIXATION FOR COW MILK CELL COUNT BY COULTER COUNTER B. POUTREL G. DUBRAY INRA

  6. Somatic Cell Counts and Infection Rates for Cows of Varying Somatic Cell Count in Initial Test of First Lactation

    Microsoft Academic Search

    E. M. COFFEYfl; W. E. Vinson; R. E. Pearson

    1986-01-01

    Somatic cell counts (log, base 2)and rates of infection in first and subsequent lactations were examined by classes of somatic cell count in initial test day of first lactation to determine if cows with initially low\\

  7. Differential Leukocyte Count Method for Bovine Low Somatic Cell Count Milk

    Microsoft Academic Search

    H. Dosogne; F. Vangroenweghe; J. Mehrzad; A. M. Massart-Leën; C. Burvenich

    2003-01-01

    Whereas many differential leukocyte count methods for high somatic cell count (SCC) milk from mastitic cows are available, only a few have been developed for low SCC milk. We have developed a flow cytometric differential leukocyte count method for low SCC milk. The procedure consists of 1) 1.5 ml of diluted milk sam- ple (30%, vol\\/vol dilution with PBS), 2)

  8. Counting

    NSDL National Science Digital Library

    Mrs. Beck

    2006-12-08

    Students will practice counting different objects. Have fun counting with this counting game. Play the game three times. Go under the sea with Fishy Count. Play the game three times. These spooky ghosts want you to practice counting by 2 s. ...

  9. Counting.

    ERIC Educational Resources Information Center

    Thwaites, G. N.

    1989-01-01

    Discusses a counting system and number operations. Suggests six distinct areas in a "number" subject: one-to-one correspondences; simple counting process; complicated counting process; addition and multiplication; algorithms for the operations; and the decimal system. (YP)

  10. Electrical cell counting process characterization in a microfluidic impedance cytometer.

    PubMed

    Hassan, Umer; Bashir, Rashid

    2014-10-01

    Particle counting in microfluidic devices with coulter principle finds many applications in health and medicine. Cell enumeration using microfluidic particle counters is fast and requires small volumes of sample, and is being used for disease diagnostics in humans and animals. A complete characterization of the cell counting process is critical for accurate cell counting especially in complex systems with samples of heterogeneous population interacting with different reagents in a microfluidic device. In this paper, we have characterized the electrical cell counting process using a microfluidic impedance cytometer. Erythrocytes were lysed on-chip from whole blood and the lysing was quenched to preserve leukocytes which subsequently pass through a 15 ?m?×?15 ?m measurement channel used to electrically count the cells. We show that cell counting over time is a non-homogeneous Poisson process and that the electrical cell counts over time show the log-normal distribution, whose skewness can be attributed to diffusion of cells in the buffer that is used to meter the blood. We further found that the heterogeneous cell population (i.e. different cell types) shows different diffusion characteristics based on the cell size. Lymphocytes spatially diffuse more as compared to granulocytes and monocytes. The time difference between the cell occurrences follows an exponential distribution and when plotted over time verifies the cell diffusion characteristics. We also characterized the probability of occurrence of more than one cell at the counter within specified time intervals using Poisson counting statistics. For high cell concentration samples, we also derived the required sample dilution based on our particle counting characterization. Buffer characterization by considering the size based particle diffusion and estimating the required dilution are critical parameters for accurate counting results. PMID:24898912

  11. Epidemiologic Considerations in Reporting Herd Somatic Cell Counts

    Microsoft Academic Search

    W. D. Hueston; L. E. Heider

    1986-01-01

    The reporting of herd summaries of individual cow somatic cell counts differs between the nine Dairy Records Pro- cessing Centers. The objectives of this paper are 1) to review the present re- porting; 2) to compare the methods used to calculate herd somatic cell counts; and 3) to discuss the epidemio- logic implications of these methods. Estimates of central tendency

  12. Heritabilities of Measures of Somatic Cell Count per Lactation

    Microsoft Academic Search

    H. G. Monardes; B. W. Kennedy; J. E. Moxley

    1983-01-01

    Measures per lactation of somatic cell count were developed from monthly test- day observations for 3,966 Holstein cows on the official test option of the Quebec Dairy Herd Analysis Service. Cows were daughters of 99 sires that had five or more daughters in two or more herds. Two lactation periods were considered. One was based on all somatic cell counts

  13. Cell Counts in Cerebral Cortex of an Autistic Patient.

    ERIC Educational Resources Information Center

    Coleman, Paul D.; And Others

    1985-01-01

    Numbers of neurons and glia were counted in the cerebral cortex of one case of autism and two age- and sex-matched controls. Cell counts were made in primary auditory cortex, Broca's speech area, and auditory association cortex. No consistent differences in cell density were found between brains of autistic and control patients. (Author/CL)

  14. Optimization of direct cell counting in sediment

    Microsoft Academic Search

    Heidi L Gough; David A Stahl

    2003-01-01

    This study reports a method for optimizing direct counts of bacteria in sediment, designed to reduce the masking by sediment particles. The protocol was designed to determine appropriate dilution factors by incorporating counting statistics and was used to measure depth-associated changes in microbial abundance in metal-impacted freshwater sediments. We demonstrated a direct method to determine appropriate sample dilution for accurate

  15. Counting unstained, confluent cells by modified bright-field microscopy

    PubMed Central

    Drey, L. Louis; Graber, Michael C.; Bieschke, Jan

    2013-01-01

    We present a very simple procedure yielding high-contrast images of adherent, confluent cells such as human neuroblastoma (SH-EP) cells by ordinary bright-field microscopy. Cells are illuminated through a color filter and a pinhole aperture placed between the condenser and the cell culture surface. Refraction by each cell body generates a sharp, bright spot when the image is defocused. The technique allows robust, automatic cell counting from a single bright-field image in a wide range of focal positions; it does this via free, readily available image-analysis tools. Contrast may be enhanced by swelling cell bodies by brief incubation in PBS. The procedure was benchmarked against manual counting and automated counting of fluorescently labeled cell nuclei.. Counts from day-old and freshly seeded plates were compared in a range of densities, from sparse to densely overgrown. On average bright-field images produced the same counts as fluorescent images, with less than 5% error. This method will allow routine cell counting using a plain bright-field microscope, absent cell-line modification or cell staining. PMID:23834382

  16. Impact of Early Lactation Somatic Cell Count in Heifers on Somatic Cell Counts Over the First Lactation

    Microsoft Academic Search

    S. De Vliegher; H. W. Barkema; H. Stryhn; G. Opsomer; A. de Kruif

    2004-01-01

    The objective of this study was to estimate the impact of somatic cell count in early lactation (SCCel) from Belgian dairy heifers on test-day somatic cell count (SCC) in first lactation. Geometric mean SCCel (5 to 14 d in milk (DIM)) of the 14,766 available samples was 104,000 cells\\/mL, and decreased from 178,000 at 5 DIM to 74,000 cells\\/mL at

  17. Fluid Flow in Cell Printing

    NASA Astrophysics Data System (ADS)

    Jalaal, Maziyar; Cheng, Eric; Ahmadi, Ali; Cheung, Karen; Stoeber, Boris

    2013-11-01

    Inkjet drop-on-demand (DOD) dispensing of cells has numerous applications including cell-based assays and tissue engineering. In our experiments, using a transparent inkjet nozzle, high speed camera, and a shadowgraphy technique, we have observed three different characteristic cell behaviors during droplet ejection: 1) traveling toward the nozzle tip, 2) ejection from the nozzle, and 3) reflection away from the nozzle tip, where the reflection is an unwanted effect which contributes to the unpredictability of current cell printing systems. To understand the reflection mechanisms, we use numerical simulation to resolve the fluid motion inside the nozzle in presence of a cell during drop formation. For this purpose an adaptive finite volume method is employed. To track the interfaces (cell-liquid, gas-liquid) a volume of fluid (VOF) method is used, where the cell is modeled as an immiscible fluid droplet with different physical properties from the suspending fluid. It is shown that after a short period of time, a recirculation zone close to the nozzle tip is generated due to droplet pinch-off. This causes a reverse flow (velocity away from the nozzle) in the center of the nozzle. This dynamic flow field inside the nozzle causes a cell to show one of the three behaviors described above depending on its initial position. Moreover, it is shown that, depending on the size, deformability, and location of the cell, the drop formation process may be influenced.

  18. DIFFERENTIAL BLOOD CELL COUNTS OF ATLANTIC HERRING, CLUPEA HARENGUS HARENGUS

    E-print Network

    DIFFERENTIAL BLOOD CELL COUNTS OF ATLANTIC HERRING, CLUPEA HARENGUS HARENGUS STUART W. SHERBURNE1 ABSTRACT In differential blood counts of 200 herring, Clupea haretll(us haretll(us. the percentages reported in the literature. Herring were sampled from February 1969 through July 1969 from the Boothbay

  19. The cytospin technique improves the detection of calcium pyrophosphate crystals in synovial fluid samples with a low leukocyte count.

    PubMed

    Robier, Christoph; Quehenberger, Franz; Neubauer, Manfred; Stettin, Mariana; Rainer, Franz

    2014-06-01

    In synovial fluids (SF) with low leukocyte or/and crystal counts, important features may be missed, if exclusively smears are examined by polarized microscopy. That may be overcome by cytocentrifuges, which use low-speed centrifugal force to concentrate cells onto a glass slide and thus enhance the number of cells per high power field (HPF). We compared the calcium pyrophosphate (CPP) crystal counts in cytospin preparations with those in common smears of SF. The number of CPP crystals was counted in 50 SF samples by polarized microscopy, and statistical comparisons of the mean values of the cytospin and smear preparations were performed using the Wilcoxon test. The reproducibility within the slides of the cytocentrifuge and smear samples was determined by Spearman's rank correlation. The crystal counts were significantly higher in the cytospin than in the smear preparations (median 96/10 HPF vs. 2.5/10 HPF, p < 0.0001). The correlation in the crystal count between the slides 1 and 2 was significantly higher within the cytocentrifuge than in the smear group (0.97 vs. 0.73, p = 0.0004). CPP-negative cytospin preparations in initially smear-positive slides were not observed. We confirmed that the cytospin technique significantly enhances the number of examinable crystals per HPF, compared to common smears. PMID:23388697

  20. Somatic cells count in cow's bulk tank milk.

    PubMed

    Olechnowicz, Jan; Ja?kowski, Jedrzej M

    2012-06-01

    The objective of this study was therefore to present factors affecting somatic cell counts in bovine bulk milk as a result of intramammary infections as well as non-infectious factors. The paper presents also the impact of on-farm management practices on the level of bulk milk somatic cell counts and presents quality indicators in bulk tank milk. At the farm level bulk milk bacterial infection takes place through three main sources: bacterial contamination from the external surface of the udder and teats, from the surface of the milking equipment, and from mastitis microorganisms within the udder. The threshold of 200,000 cells/ml identifies bacteriological negative quarters of the udder. The counts of mammary pathogens in bulk tank milk are relatively low, on average not exceeding 1,000 cfu/ml. Environmental pathogens predominate in bulk tank milk samples with somatic cells count <300 × 10(3) ml. PMID:22230979

  1. Sample to answer visualization pipeline for low-cost point-of-care blood cell counting

    NASA Astrophysics Data System (ADS)

    Smith, Suzanne; Naidoo, Thegaran; Davies, Emlyn; Fourie, Louis; Nxumalo, Zandile; Swart, Hein; Marais, Philip; Land, Kevin; Roux, Pieter

    2015-03-01

    We present a visualization pipeline from sample to answer for point-of-care blood cell counting applications. Effective and low-cost point-of-care medical diagnostic tests provide developing countries and rural communities with accessible healthcare solutions [1], and can be particularly beneficial for blood cell count tests, which are often the starting point in the process of diagnosing a patient [2]. The initial focus of this work is on total white and red blood cell counts, using a microfluidic cartridge [3] for sample processing. Analysis of the processed samples has been implemented by means of two main optical visualization systems developed in-house: 1) a fluidic operation analysis system using high speed video data to determine volumes, mixing efficiency and flow rates, and 2) a microscopy analysis system to investigate homogeneity and concentration of blood cells. Fluidic parameters were derived from the optical flow [4] as well as color-based segmentation of the different fluids using a hue-saturation-value (HSV) color space. Cell count estimates were obtained using automated microscopy analysis and were compared to a widely accepted manual method for cell counting using a hemocytometer [5]. The results using the first iteration microfluidic device [3] showed that the most simple - and thus low-cost - approach for microfluidic component implementation was not adequate as compared to techniques based on manual cell counting principles. An improved microfluidic design has been developed to incorporate enhanced mixing and metering components, which together with this work provides the foundation on which to successfully implement automated, rapid and low-cost blood cell counting tests.

  2. Proteolysis in Milk Associated with Increasing Somatic Cell Counts

    Microsoft Academic Search

    G. F. Senyk; D. M. Barbano; W. F. Shipe

    1985-01-01

    Individual cow samples were collected and preserved with potassium dichromate. Somatic cells counts were determined. Tyrosine value was used as an index of pro- teolysis. Sixty-six samples ranged in somatic cell count from < 50,000 to > 2,000,000\\/ml. Initial milk tyrosine values and tyrosine values for milks incubated for 24 h at 37°C showed proteolytic activity increased with increasing somatic

  3. Automating Single Subunit Counting of Membrane Proteins in Mammalian Cells*

    PubMed Central

    McGuire, Hugo; Aurousseau, Mark R. P.; Bowie, Derek; Blunck, Rikard

    2012-01-01

    Elucidating subunit stoichiometry of neurotransmitter receptors is preferably carried out in a mammalian expression system where the rules of native protein assembly are strictly obeyed. Although successful in Xenopus oocytes, single subunit counting, manually counting photobleaching steps of GFP-tagged subunits, has been hindered in mammalian cells by high background fluorescence, poor control of expression, and low GFP maturation efficiency. Here, we present a fully automated single-molecule fluorescence counting method that separates tagged proteins on the plasma membrane from background fluorescence and contaminant proteins in the cytosol or the endoplasmic reticulum and determines the protein stoichiometry. Lower GFP maturation rates observed in cells cultured at 37 °C were partly offset using a monomeric version of superfolder GFP. We were able to correctly identify the stoichiometry of GluK2 and ?1 glycine receptors. Our approach permits the elucidation of stoichiometry for a wide variety of plasma membrane proteins in mammalian cells with any commercially available TIRF microscope. PMID:22930752

  4. Dynamics and regulation of bulk milk somatic cell counts.

    PubMed Central

    Schukken, Y H; Weersink, A; Leslie, K E; Martin, S W

    1993-01-01

    Somatic cell count (SCC) in milk is inversely related to dairy cow productivity and milk quality. In an effort to improve product quality, and indirectly farm productivity, regulatory limits on somatic cell counts have been established by many of the major dairy producing countries. The purpose of this paper was to assess the impact of regulations on bulk milk somatic cell counts in Ontario and to assist producers in meeting regulatory limits through development of prediction models. Through the use of a transfer function model, provincial SCC was found to have dropped by approximately 60,000 as a result of the reduction program. Limits of the regulatory program, seasonality and herd characteristics were found through time series cross-sectional models to have an impact on prediction of SCC at the farm level, but the major influence was historical SCC levels. PMID:8490807

  5. Tennessee Quality Milk Initiative Using Milk Somatic Cell Count Information

    Microsoft Academic Search

    Kristy H. Campbell

    The level of mastitis infection in a dairy herd can have a significant impact on herd profitability. Losses due to mastitis include decreased milk production, increased treatment costs, dis- carded milk, premature culling, death, decreased genetic potential, decreased reproductive performance, load rejection due to violation of somatic cell counts (SCC) or antibiotic resi- dues and loss of milk quality premiums

  6. Prognostic Value of Elevated White Blood Cell Count in Hypertension

    Microsoft Academic Search

    Giuseppe Schillaci; Matteo Pirro; Giacomo Pucci; Tiziana Ronti; Gaetano Vaudo; Massimo R. Mannarino; Carlo Porcellati; Elmo Mannarino

    2007-01-01

    Background: Chronic low-grade inflammation may contribute to vascular injury and atherogenesis, and has been described in association to high blood pressure (BP). However, as yet the prognostic significance of white blood cell (WBC) count in the setting of uncomplicated hypertension has not been investigated.Methods: In the Progetto Ipertensione Umbria Monitoraggio Ambulatoriale (PIUMA) study, 1617 white patients with essential hypertension (aged

  7. Effect of Freezing on Fossomatic Cell Counting in Ewe Milk

    Microsoft Academic Search

    J. R. Martínez; C. Gonzalo; J. A. Carriedo; F. San Primitivo

    2003-01-01

    Using the Fossomatic method, a total of 10,072 ana- lytical somatic cell count (SCC) observations were car- ried out on 4760 aliquots taken from 70 individual ewe milk samples with the objective of studying whether freezing showed significant differences of SCC in com- parison with refrigeration, according to different ana- lytical conditions. These conditions were four preserva- tion procedures (without

  8. Effect of Clinical Contagious Agalactia on the Bulk Tank Milk Somatic Cell Count in Murciano-Granadina Goat Herds

    Microsoft Academic Search

    J. C. Corrales; A. Sánchez; C. Luengo; J. B. Poveda; A. Contreras

    2004-01-01

    From 19 herds of Murciano-Granadina goats, weekly bulk tank somatic cell count (BTSCC) were performed from October to April, and suspicious milk (n = 182), synovial fluid, and ocular swabs (n = 15) from diseased goats were processed for mycoplasma isolation and identification. Also BTSCC from 65 herds were deter- mined (n = 2693). A mixed model procedure was used

  9. An optical counting technique with vertical hydrodynamic focusing for biological cells

    PubMed Central

    Chiavaroli, Stefano; Newport, David; Woulfe, Bernie

    2010-01-01

    A barrier in scaling laboratory processes into automated microfluidic devices has been the transfer of laboratory based assays: Where engineering meets biological protocol. One basic requirement is to reliably and accurately know the distribution and number of biological cells being dispensed. In this study, a novel optical counting technique to efficiently quantify the number of cells flowing into a microtube is presented. REH, B-lymphoid precursor leukemia, are stained with a fluorescent dye and frames of moving cells are recorded using a charge coupled device (CCD) camera. The basic principle is to calculate the total fluorescence intensity of the image and to divide it by the average intensity of a single cell. This method allows counting the number of cells with an uncertainty ±5%, which compares favorably to the standard biological methodology, based on the manual Trypan Blue assay, which is destructive to the cells and presents an uncertainty in the order of 20%. The use of a microdevice for vertical hydrodynamic focusing, which can reduce the background noise of out of focus cells by concentrating the cells in a thin layer, has further improved the technique. Computational fluid dynamics (CFD) simulation and confocal laser scanning microscopy images have shown an 82% reduction in the vertical displacement of the cells. For the flow rates imposed during this study, a throughput of 100–200 cells?s is achieved. PMID:20697579

  10. Predicting Somatic Cell Count Standard Violations Based on Herd's Bulk Tank Somatic Cell Count. Part II: Consistency Index

    Microsoft Academic Search

    J. M. Lukas; J. K. Reneau; C. Munoz-Zanzi; M. L. Kinsel

    2008-01-01

    The present study examines the capability of 1,501 herds in the Upper Midwest and the performance of statistical process control charts and indices as a way of monitoring and controlling milk quality on the farm. For 24 mo, daily or every other day bulk tank somatic cell count (SCC) data were collected. Consistency indi- ces for 5 different SCC standards

  11. The association between high milk somatic cell counts in the first lactation and somatic cell counts in the second lactation

    Microsoft Academic Search

    H. J. Williams; P. J. Cripps; D. H. Grove-White

    With the advent of web-based recording and analysis systems, individual cow composite somatic cell count (SCC) data are being increasingly used for decision support in mastitis control at both the individual cow and herd level. SCC data from first and second lactation dairy cows (n=1912) from 12 farms were analysed using multinomial logistic regression to investigate possible associations between high

  12. Laser rastering flow cytometry: fast cell counting and identification

    NASA Astrophysics Data System (ADS)

    Vacca, G.; Junnarkar, M. R.; Goldblatt, N. R.; Yee, M. W.; Van Slyke, B. M.; Briese, T. C.

    2009-02-01

    We describe the concept of laser rastering flow cytometry, where a rapidly scanning laser beam allows counting and classification of cells at much higher rates than currently possible. Modifications to existing flow cytometers to implement the concept include an acousto-optic deflector, fast analog-to-digital conversion, and a two-step digital-signal-processing scheme that handles the high data rates and provides key assay information. Results are shown that prove the concept, demonstrating the ability to resolve closely spaced cells and to measure cells at rates more than an order of magnitude faster than on conventional flow-cytometer-based hematology analyzers.

  13. Intracellular fluid flow in rapidly moving cells.

    PubMed

    Keren, Kinneret; Yam, Patricia T; Kinkhabwala, Anika; Mogilner, Alex; Theriot, Julie A

    2009-10-01

    Cytosolic fluid dynamics have been implicated in cell motility because of the hydrodynamic forces they induce and because of their influence on transport of components of the actin machinery to the leading edge. To investigate the existence and the direction of fluid flow in rapidly moving cells, we introduced inert quantum dots into the lamellipodia of fish epithelial keratocytes and analysed their distribution and motion. Our results indicate that fluid flow is directed from the cell body towards the leading edge in the cell frame of reference, at about 40% of cell speed. We propose that this forward-directed flow is driven by increased hydrostatic pressure generated at the rear of the cell by myosin contraction, and show that inhibition of myosin II activity by blebbistatin reverses the direction of fluid flow and leads to a decrease in keratocyte speed. We present a physical model for fluid pressure and flow in moving cells that quantitatively accounts for our experimental data. PMID:19767741

  14. Correlation between standard plate count and somatic cell count milk quality results for Wisconsin dairy producers.

    PubMed

    Borneman, Darand L; Ingham, Steve

    2014-05-01

    The objective of this study was to determine if a correlation exists between standard plate count (SPC) and somatic cell count (SCC) monthly reported results for Wisconsin dairy producers. Such a correlation may indicate that Wisconsin producers effectively controlling sanitation and milk temperature (reflected in low SPC) also have implemented good herd health management practices (reflected in low SCC). The SPC and SCC results for all grade A and B dairy producers who submitted results to the Wisconsin Department of Agriculture, Trade, and Consumer Protection, in each month of 2012 were analyzed. Grade A producer SPC results were less dispersed than grade B producer SPC results. Regression analysis showed a highly significant correlation between SPC and SCC, but the R(2) value was very small (0.02-0.03), suggesting that many other factors, besides SCC, influence SPC. Average SCC (across 12 mo) for grade A and B producers decreased with an increase in the number of monthly SPC results (out of 12) that were ? 25,000 cfu/mL. A chi-squared test of independence showed that the proportion of monthly SCC results >250,000 cells/mL varied significantly depending on whether the corresponding SPC result was ? 25,000 or >25,000 cfu/mL. This significant difference occurred in all months of 2012 for grade A and B producers. The results suggest that a generally consistent level of skill exists across dairy production practices affecting SPC and SCC. PMID:24630657

  15. Reference system for somatic cell counting in milk

    Microsoft Academic Search

    Silvia Orlandini; Harrie van den Bijgaart

    2011-01-01

    With some analytical parameters, certified reference materials are lacking and the reference method shows limited performance.\\u000a Somatic cell counting in milk is a clear example. It is one of the most frequently performed measurements, estimated at over\\u000a 500 000 000 tests\\/year world wide. It serves as an indicator for the udder health status of lactating animals, is relevant\\u000a in food legislation,

  16. Predictability by Somatic Cell Counts Related to Prevalence of Intrammary Infection Within Herds

    Microsoft Academic Search

    M. P. McDermott; H. N. Erb; R. P. Natzke

    1982-01-01

    Somatic cells were counted and bacteria identified for milk samples from 719 lactating dairy cows in 12 commercial herds. These pooled data were used to look at the accuracy of alternative thresholds of somatic cell counts as indicators of intramammary infection. Sensitivity, specificity, and predictability positive and negative at alternative cell count thresholds were calculated. There was an increase of

  17. Mastitis Therapy for Cows with Elevated Somatic Cell Counts or Clinical Mastitis1

    Microsoft Academic Search

    L. L. Timms; L. H. Schultz

    1984-01-01

    Intramammary treatment with a broad spectrum antibiotic was evaluated for cows treated after a single high monthly somatic cell count or for cows with clinical mastitis. Forty-three quarters of 36 cows were treated after a high somatic cell count, and 56 quarters of 48 cows were treated after clinical symptoms. There was no significant decrease of cell count in response

  18. A cell number-counting factor regulates the cytoskeleton and cell motility in Dictyostelium

    Microsoft Academic Search

    Lei Tang; Tong Gao; Catherine McCollum; Wonhee Jang; Michael G. Vicker; Robin R. Ammann; Richard H. Gomer

    2002-01-01

    Little is known about how a morphogenetic rearrangement of a tissue is affected by individual cells. Starving Dictyostelium discoideum cells aggregate to form dendritic streams, which then break up into groups of 2 × 104 cells. Cell number is sensed at this developmental stage by using counting factor (CF), a secreted complex of polypeptides. A high extracellular concentration of CF

  19. Cell Volume to Aid Analysis and Technique of Somatic Cell Counts in Milk

    Microsoft Academic Search

    R. F. Sheldrake; R. J. T. Hoare; V. E. Woodhouse; G. D. McGregor

    1977-01-01

    In conjunction with a Coulter Coun- ter, somatic cells in milk were sized by electronic analysis. Quarter milk from cows with mastitis had a cell volume peak with a modal cell volume of 102\\/23 while milk from healthy quarters had no peak. Bulk milks with a peak had higher cell counts than milks where there was no peak. Dimensions of

  20. Cell detection and counting through cell lysate impedance spectroscopy in microfluidic devices†

    PubMed Central

    Cheng, Xuanhong; Liu, Yi-shao; Irimia, Daniel; Demirci, Utkan; Yang, Liju; Zamir, Lee; Rodríguez, William R.; Toner, Mehmet; Bashir, Rashid

    2015-01-01

    Cell-based microfluidic devices have attracted interest for a wide range of applications. While optical cell counting and flow cytometry-type devices have been reported extensively, sensitive and efficient non-optical methods to detect and quantify cells attached over large surface areas within microdevices are generally lacking. We describe an electrical method for counting cells based on the measurement of changes in conductivity of the surrounding medium due to ions released from surface-immobilized cells within a microfluidic channel. Immobilized cells are lysed using a low conductivity, hypotonic media and the resulting change in impedance is measured using surface patterned electrodes to detect and quantify the number of cells. We found that the bulk solution conductance increases linearly with the number of isolated cells contributing to solution ion concentration. The method of cell lysate impedance spectroscopy is sensitive enough to detect 20 cells ?L–1, and offers a simple and efficient method for detecting and enumerating cells within microfluidic devices for many applications including measurement of CD4 cell counts in HIV patients in resource-limited settings. To our knowledge, this is the most sensitive approach using non-optical setups to enumerate immobilized cells. The microfluidic device, capable of isolating specific cell types from a complex bio-fluidic and quantifying cell number, can serve as a single use cartridge for a hand-held instrument to provide simple, fast and affordable cell counting in point-of-care settings. PMID:17538717

  1. Comparison of Milk Somatic Cell Counts by Coulter and Fossomatic Counters1

    Microsoft Academic Search

    R. H. Miller; M. J. Paape; J. C. Acton

    1986-01-01

    Unpreserved milk samples from 28 quarters of 18 cows were used to compare milk somatic cell counts obtained by Fossomatic and Coulter Counter and to determine effect of temperature and sample age on Fossomatic counts. Samples represented high and low cell count milk (16 cows) and colostrum (2 cows). Fifteen milliliters of both foremilk (after milking preparation) and strippings were

  2. Normal values for peripheral blood white cell counts in women of four different ethnic origins

    Microsoft Academic Search

    B Bain; M Seed; I Godsland

    1984-01-01

    Total and differential white cell counts were studied in 399 women living in the same community in Britain but drawn from four different ethnic groups. The groups were white (northern European), Indian, black (African and West Indian), and Oriental. The total white cell count and absolute neutrophil count were significantly lower in the black group than in each of the

  3. Prediction of bulk tank somatic cell count violations based on monthly individual cow somatic cell count data.

    PubMed

    Fauteux, V; Bouchard, E; Haine, D; Scholl, D T; Roy, J P

    2015-04-01

    The regulatory limit in Canada for bulk tank somatic cell count (BTSCC) was recently lowered from 500,000 to 400,000 cells/mL. Herd indices based on changes in cow somatic cell count over 2 consecutive months (e.g., proportion of healthy or chronically infected cows, cows cured, and new intramammary infection rate) could be used as predictors for BTSCC violations. The objective of this study was to develop a predictive model for exceeding the limit of 400,000 cells/mL in the next month using these herd indices. Dairy Herd Improvement (DHI) data were used from 924 dairy herds in Québec, Canada. Test-day BTSCC was estimated by dividing the sum of all cows' DHI test-day somatic cell count times DHI test-day milk production by the total volume of milk produced by the herd on that test-day. In total, 986 of 8,681 (11.4%) estimated BTSCC exceeded 400,000 cells/mL. The final predictive model included 6 variables: mean herd somatic cell score at the current test-month, proportion of cows >500,000 cells/mL at the current test-month, proportion of healthy cows during lactation at the current test-month, proportion of chronically infected cows at the current test-month, average days in milk at the current test-month, and annual mean daily milk production. The optimized sensitivity and specificity of the model were 76 and 74%, respectively. The positive predictive value and negative predictive value were 25 and 95%, respectively. This low positive predictive value and high negative predictive value demonstrated that the model was less accurate at predicting herds that would violate the estimated BTSCC threshold but very accurate at identifying herds that would not. In addition, the area under the curve for the receiver operating characteristic curve was 0.82, suggesting that the model had excellent discrimination between test-months that did and did not exceed 400,000 cells/mL. An internal validation was completed using a bootstrapped resampling-based estimation method and confirmed that the final model provided a validated estimate of predictive accuracy. This model could be used to monitor and advise clients on impending risks of exceeding the BTSCC limit. PMID:25704970

  4. Use of domestic detergents in the California mastitis test for high somatic cell counts in milk.

    PubMed

    Leach, K A; Green, M J; Breen, J E; Huxley, J N; Macaulay, R; Newton, H T; Bradley, A J

    2008-11-01

    The California mastitis test (CMT) is used on farms to identify subclinical mastitis by an indirect estimation of the somatic cell count (SCC) in milk. Four commercially available detergents were compared with a bespoke cmt fluid for their ability to detect milk samples with a scc above 200,000 cells/ml; differences between the interpretation of the results of the tests by eight operators were also investigated. The sensitivity and specificity of the test were affected by the type of detergent, and by the operators' interpretations. When used by the most sensitive operator, suitably diluted Fairy Liquid performed almost identically to cmt fluid in identifying milk samples with more than 200,000 cells/ml. The average sensitivities achieved by the eight operators for detecting this threshold were 82 per cent for Fairy Liquid and 84 per cent for cmt fluid, and the specificities were 93 and 91 per cent respectively. The other detergents contained less anionic surfactants and were less sensitive but similarly specific. PMID:18997186

  5. CELLCOUNTER: Novel Open-Source Software for Counting Cell Migration and Invasion In Vitro

    PubMed Central

    Yang, Hongshun; Zhu, Tao

    2014-01-01

    Transwell Boyden chamber based migration/invasion assay is a simple and extensively used approach for the characterization of cell motility in vitro. Cell motility is quantified by counting the number of cells that pass through the filter membrane. The counting is usually performed manually, which is laborious and error prone. We have therefore developed CELLCOUNTER, an application that is capable of recognizing and counting the total number of cells through an intuitive graphical user interface. The counting can be performed in batch, and the counting results can be visualized and further curated manually. CELLCOUNTER will be helpful in streamlining the experimental process and improving the reliability of the data acquisition. PMID:25054152

  6. Sensitive fluorometric nanoparticle assays for cell counting and viability.

    PubMed

    Pihlasalo, Sari; Pellonperä, Lotta; Martikkala, Eija; Hänninen, Pekka; Härmä, Harri

    2010-11-15

    We have developed easy-to-use homogeneous methods utilizing time-resolved fluorescence resonance energy transfer (TR-FRET) and fluorescence quenching for quantification of eukaryotic cells. The methods rely on a competitive adsorption of cells and fluorescently labeled protein onto citrate-stabilized colloidal gold nanoparticles or carboxylate-modified polystyrene nanoparticles doped with an Eu(III) chelate. In the gold nanoparticle sensor, the adsorption of the labeled protein to the gold nanoparticles leads to quenching of the fluorochrome. Eukaryotic cells reduce the adsorption of labeled protein to the gold particles increasing the fluorescence signal. In the Eu(III) nanoparticle sensor, the time-resolved fluorescence resonance energy transfer between the nanoparticles and an acceptor-labeled protein is detected; a decrease in the magnitude of the time-resolved energy transfer signal (sensitized time-resolved fluorescence) is proportional to the cell-nanoparticle interaction and subsequent reduced adsorption of the labeled protein. Less than five cells were detected and quantified with the nanoparticle sensors in the homogeneous microtiter assay format with a coefficient of variation of 6% for the gold and 12% for the Eu(III) nanoparticle sensor. The Eu(III) nanoparticle sensor was also combined with a cell impermeable nucleic acid dye assay to measure cell viability in a single tube test with cell counts below 1000 cells/tube. This sensitive and easy-to-use nanoparticle sensor combined with a viability test for a low concentration of cells could potentially replace existing microscopic methods in biochemical laboratories. PMID:20954745

  7. Counting Legionella cells within single amoeba host cells

    EPA Science Inventory

    Here we present the first attempt to quantify L. pneumophila cell numbers within individual amoebae hosts that may be released into engineered water systems. The maximum numbers of culturable L. pneumophila cells grown within Acanthamoeba polyphaga and Naegleria fowleri were 134...

  8. Counting Legionella cells within single amoeba host cells.

    PubMed

    Buse, Helen Y; Ashbolt, Nicholas J

    2012-03-01

    Here we present the first attempt to quantify Legionella pneumophila cell numbers within individual amoeba hosts that may be released into engineered water systems. The maximum numbers of culturable L. pneumophila cells grown within Acanthamoeba polyphaga and Naegleria fowleri were 1,348 (mean, 329) and 385 (mean, 44) CFU trophozoite(-1), respectively. PMID:22226955

  9. Use of somatic cell counts for the detection of subclinical mastitis in sheep

    Microsoft Academic Search

    A. P. Mavrogenis; A. Koumas; C. K. Kakoyiannis; C. H. Taliotis

    1995-01-01

    A study using 100 Chios ewes was undertaken in 1992 to determine rate of clinical mastitis, prevalent bacterial pathogens in normal and abnormal milk and to investigate relationships between somatic cell counts and milk production. Mean somatic cell count from mastitis negative (non-infected) samples was 1.574 × 106 cells per ml?1. All mastitis positive samples had somatic cells in excess

  10. Bulk milk somatic cell counts are related to bulk milk total bacterial counts and several herd-level risk factors in dairy goats

    Microsoft Academic Search

    G. Koop; M. Nielen; T. van Werven

    2009-01-01

    The aim of this study was to describe the temporal variation in bulk milk somatic cell count (BMSCC) on Dutch dairy goat farms and to assess the correla- tion of BMSCC with bulk milk total bacterial counts (BMTBC) and with several herd management factors. Bulk milk somatic cell count and BMTBC data were recorded from 90% of the dairy goat

  11. Intracellular fluid flow in rapidly moving cells

    PubMed Central

    Keren, Kinneret; Yam, Patricia T.; Kinkhabwala, Anika; Mogilner, Alex; Theriot, Julie A.

    2010-01-01

    Cytosolic fluid dynamics have been implicated in cell motility1–5 because of the hydrodynamic forces they induce and because of their influence on transport of components of the actin machinery to the leading edge. To investigate the existence and the direction of fluid flow in rapidly moving cells, we introduced inert quantum dots into the lamellipodia of fish epithelial keratocytes and analysed their distribution and motion. Our results indicate that fluid flow is directed from the cell body towards the leading edge in the cell frame of reference, at about 40% of cell speed. We propose that this forward-directed flow is driven by increased hydrostatic pressure generated at the rear of the cell by myosin contraction, and show that inhibition of myosin II activity by blebbistatin reverses the direction of fluid flow and leads to a decrease in keratocyte speed. We present a physical model for fluid pressure and flow in moving cells that quantitatively accounts for our experimental data. PMID:19767741

  12. Effect of Streess on Blood Leucocyte and Milk Somatic Cell Counts in Dairy Cows1

    Microsoft Academic Search

    T. N. Wegner; J. D. Schuh; F. E. Nelson; G. H. Stott

    1976-01-01

    Blood and milk samples from Holstein cows were examined for total blood leucocyte count, differential blood leuco- cyte count, milk quality test, and somatic cell count in milk while the cows were stressed by corticotropin injection, con- finement in a heat-humidity chamber, or environmental-heat stress by exposure during the hot summer months of June through November in southern Arizona. All

  13. FISH and Chips: Automation of Fluorescent Dot Counting in Interphase Cell Nuclei

    E-print Network

    van Vliet, Lucas J.

    in situ hybridization (FISH) techniques in interphase cell nuclei have great potential, both in re- searchFISH and Chips: Automation of Fluorescent Dot Counting in Interphase Cell Nuclei Hans Netten,1 Ian abnormalities in inter- phase cell nuclei. This process is called dot counting. To estimate the distribution

  14. Determination of mammalian cell counts, cell size and cell health using the Moxi Z mini automated cell counter.

    PubMed

    Dittami, Gregory M; Sethi, Manju; Rabbitt, Richard D; Ayliffe, H Edward

    2012-01-01

    Particle and cell counting is used for a variety of applications including routine cell culture, hematological analysis, and industrial controls(1-5). A critical breakthrough in cell/particle counting technologies was the development of the Coulter technique by Wallace Coulter over 50 years ago. The technique involves the application of an electric field across a micron-sized aperture and hydrodynamically focusing single particles through the aperture. The resulting occlusion of the aperture by the particles yields a measurable change in electric impedance that can be directly and precisely correlated to cell size/volume. The recognition of the approach as the benchmark in cell/particle counting stems from the extraordinary precision and accuracy of its particle sizing and counts, particularly as compared to manual and imaging based technologies (accuracies on the order of 98% for Coulter counters versus 75-80% for manual and vision-based systems). This can be attributed to the fact that, unlike imaging-based approaches to cell counting, the Coulter Technique makes a true three-dimensional (3-D) measurement of cells/particles which dramatically reduces count interference from debris and clustering by calculating precise volumetric information about the cells/particles. Overall this provides a means for enumerating and sizing cells in a more accurate, less tedious, less time-consuming, and less subjective means than other counting techniques(6). Despite the prominence of the Coulter technique in cell counting, its widespread use in routine biological studies has been prohibitive due to the cost and size of traditional instruments. Although a less expensive Coulter-based instrument has been produced, it has limitations as compared to its more expensive counterparts in the correction for "coincidence events" in which two or more cells pass through the aperture and are measured simultaneously. Another limitation with existing Coulter technologies is the lack of metrics on the overall health of cell samples. Consequently, additional techniques must often be used in conjunction with Coulter counting to assess cell viability. This extends experimental setup time and cost since the traditional methods of viability assessment require cell staining and/or use of expensive and cumbersome equipment such as a flow cytometer. The Moxi Z mini automated cell counter, described here, is an ultra-small benchtop instrument that combines the accuracy of the Coulter Principle with a thin-film sensor technology to enable precise sizing and counting of particles ranging from 3-25 microns, depending on the cell counting cassette used. The M type cassette can be used to count particles from with average diameters of 4 - 25 microns (dynamic range 2 - 34 microns), and the Type S cassette can be used to count particles with and average diameter of 3 - 20 microns (dynamic range 2 - 26 microns). Since the system uses a volumetric measurement method, the 4-25 microns corresponds to a cell volume range of 34 - 8,180 fL and the 3 - 20 microns corresponds to a cell volume range of 14 - 4200 fL, which is relevant when non-spherical particles are being measured. To perform mammalian cell counts using the Moxi Z, the cells to be counted are first diluted with ORFLO or similar diluent. A cell counting cassette is inserted into the instrument, and the sample is loaded into the port of the cassette. Thousands of cells are pulled, single-file through a "Cell Sensing Zone" (CSZ) in the thin-film membrane over 8-15 seconds. Following the run, the instrument uses proprietary curve-fitting in conjunction with a proprietary software algorithm to provide coincidence event correction along with an assessment of overall culture health by determining the ratio of the number of cells in the population of interest to the total number of particles. The total particle counts include shrunken and broken down dead cells, as well as other debris and contaminants. The results are presented in histogram format with an automatic curve fit, with gates that can

  15. Determination of Mammalian Cell Counts, Cell Size and Cell Health Using the Moxi Z Mini Automated Cell Counter

    PubMed Central

    Dittami, Gregory M.; Sethi, Manju; Rabbitt, Richard D.; Ayliffe, H. Edward

    2012-01-01

    Particle and cell counting is used for a variety of applications including routine cell culture, hematological analysis, and industrial controls1-5. A critical breakthrough in cell/particle counting technologies was the development of the Coulter technique by Wallace Coulter over 50 years ago. The technique involves the application of an electric field across a micron-sized aperture and hydrodynamically focusing single particles through the aperture. The resulting occlusion of the aperture by the particles yields a measurable change in electric impedance that can be directly and precisely correlated to cell size/volume. The recognition of the approach as the benchmark in cell/particle counting stems from the extraordinary precision and accuracy of its particle sizing and counts, particularly as compared to manual and imaging based technologies (accuracies on the order of 98% for Coulter counters versus 75-80% for manual and vision-based systems). This can be attributed to the fact that, unlike imaging-based approaches to cell counting, the Coulter Technique makes a true three-dimensional (3-D) measurement of cells/particles which dramatically reduces count interference from debris and clustering by calculating precise volumetric information about the cells/particles. Overall this provides a means for enumerating and sizing cells in a more accurate, less tedious, less time-consuming, and less subjective means than other counting techniques6. Despite the prominence of the Coulter technique in cell counting, its widespread use in routine biological studies has been prohibitive due to the cost and size of traditional instruments. Although a less expensive Coulter-based instrument has been produced, it has limitations as compared to its more expensive counterparts in the correction for "coincidence events" in which two or more cells pass through the aperture and are measured simultaneously. Another limitation with existing Coulter technologies is the lack of metrics on the overall health of cell samples. Consequently, additional techniques must often be used in conjunction with Coulter counting to assess cell viability. This extends experimental setup time and cost since the traditional methods of viability assessment require cell staining and/or use of expensive and cumbersome equipment such as a flow cytometer. The Moxi Z mini automated cell counter, described here, is an ultra-small benchtop instrument that combines the accuracy of the Coulter Principle with a thin-film sensor technology to enable precise sizing and counting of particles ranging from 3-25 microns, depending on the cell counting cassette used. The M type cassette can be used to count particles from with average diameters of 4 - 25 microns (dynamic range 2 - 34 microns), and the Type S cassette can be used to count particles with and average diameter of 3 - 20 microns (dynamic range 2 - 26 microns). Since the system uses a volumetric measurement method, the 4-25 microns corresponds to a cell volume range of 34 - 8,180 fL and the 3 - 20 microns corresponds to a cell volume range of 14 - 4200 fL, which is relevant when non-spherical particles are being measured. To perform mammalian cell counts using the Moxi Z, the cells to be counted are first diluted with ORFLO or similar diluent. A cell counting cassette is inserted into the instrument, and the sample is loaded into the port of the cassette. Thousands of cells are pulled, single-file through a "Cell Sensing Zone" (CSZ) in the thin-film membrane over 8-15 seconds. Following the run, the instrument uses proprietary curve-fitting in conjunction with a proprietary software algorithm to provide coincidence event correction along with an assessment of overall culture health by determining the ratio of the number of cells in the population of interest to the total number of particles. The total particle counts include shrunken and broken down dead cells, as well as other debris and contaminants. The results are presented in histogram format with an automatic curve fit, with gates that can be adjusted manually as

  16. Effect of parity and milk production on somatic cell count, standard plate count and composition of goat milk

    Microsoft Academic Search

    S. S. Zeng; E. N. Escobar

    1995-01-01

    Fifteen milking does were randomly selected from the Alpine herd of the Langston University farm, with five each in Parities 1, 2, and 3 (P1, P2 and P3, respectively). Composite milk samples of equal volume of morning and evening milkings were taken once a month throughout the lactation period of 8 months. All samples were analyzed for somatic cell count

  17. Temporal trends in bulk tank somatic cell count and total bacterial count in Irish dairy herds during the past decade.

    PubMed

    Berry, D P; O'Brien, B; O'Callaghan, E J; Sullivan, K O; Meaney, W J

    2006-10-01

    The objective of this study was to document temporal trends in bulk tank somatic cell count (SCC) and total bacterial counts (TBC) in Irish dairy herds during the years 1994 to 2004. Three milk processors participated in the study, providing data on 2,754,270 individual bulk tank SCC and 2,056,992 individual bulk tank TBC records from 9,113 herds. Somatic cell counts decreased during the years 1994 to 2000, followed by an annual increase thereafter of more than 2,000 cells/mL. A tendency existed for TBC to decrease over time. Across all years, bulk tank SCC were the lowest in April and highest in November; TBC were the lowest in May and highest in December. The significant seasonal pattern observed in herd SCC and TBC was an artifact of seasonal calving in Ireland. In general, herds selling more milk had lower bulk tank SCC and TBC. Herds having the highest SCC (i.e., > 450,000 cells/mL) and the lowest SCC (i.e., < or = 150,000 cells/mL) both contributed substantially to the mean SCC of the milk pool collected by the milk processors. Derived transition matrices showed that between adjacent years, herds had the greatest probability of remaining in the same annual mean SCC or TBC category. PMID:16960086

  18. Analysis of bone marrow aspiration fluid using automated blood cell counters.

    PubMed

    d'Onofrio, Giuseppe; Zini, Gina

    2015-03-01

    Cytomorphological examination of aspirate smears remains the basic method to diagnose hematologic disorders and to evaluate treatment-related changes. Last-generation hematological analyzers can count, besides cells normally circulating in peripheral blood, some types of immature and abnormal cells, such as erythroblasts and immature granulocytes. The complex nature of bone marrow fluid, however, has prevented until now the routine utilization of blood cell counters in this area. Recent studies have shown the possibility of using bone marrow fluid as a substitute for peripheral blood for clinical tests in particular situations and for repetitive cytologic examinations in specific clinical and research fields. PMID:25676370

  19. Studies on Somatic Cell Counts in Milk from Swedish Dairy Cows

    Microsoft Academic Search

    Ulf Emanuelson; Elisabeth Persson

    1984-01-01

    A program has started in Sweden to screeen the udder health status of individual cows by bucket somatic cell counting. A study was undertaken to investigate the causes of variation in test-day somatic cell counts (SCC) in milk sampled at monthly intervals. The material included data from two breeds and totalled 87760 observations. The SCC were transformed to a log

  20. Zinc Supplementation and Somatic Cell Count in Milk of Dairy Cows

    Microsoft Academic Search

    A. Pechová; L. Pavlata; E. Lokajová

    2006-01-01

    Pechová A., L. Pavlata, E. Lokajová: Zinc Supplementation and Somatic Cell Count in Milk of Dairy Cows. Acta Vet Brno 2006, 75: 355-361. The goal of the study was to test the possibility of raising milk zinc (Zn) concentration by increasing the supplementation of Zn, and to assess the effect on the somatic cell count. The experiment was performed at

  1. Studies on Somatic Cell Counts in Milk from Swedish Dairy Cows

    Microsoft Academic Search

    Ulf Emanuelson; Jan Philipsson

    1984-01-01

    A program has started in Sweden to screen the udder health status of individual cows by bucket somatic cell counting. A study was undertaken to investigate the genetic variation in test-day somatic cell counts (SCC) in milk sampled at monthly intervals. The material included data from two breeds and totalled 87760 observations. The SCC were transformed to a log scale

  2. The Effect of Season on Somatic Cell Count and the Incidence of Clinical Mastitis

    Microsoft Academic Search

    R. G. M. Olde Riekerink; H. W. Barkema; H. Stryhn

    2007-01-01

    Bulk milk somatic cell count (BMSCC), individual cow somatic cell count (ICSCC), and incidence rate of clinical mastitis (IRCM) are all udder health parame- ters. So far, no studies have been reported on the effect of season on BMSCC, IRCM, and ICSCC in the same herds and period over multiple years. The objectives of this study were to determine the

  3. Short Communication: Contribution of Vibration and Noise During Milking to the Somatic Cell Count of Milk

    Microsoft Academic Search

    L. Gygax; D. Nosal

    2006-01-01

    We investigated the hypothesis that somatic cell counts (SCC) in milk are influenced by the vibration and noise experienced by dairy cows during milking. We therefore measured vibration and noise on 50 Swiss dairy farms (with herringbone, autotandem, side-by- side, or carousel parlors), where we also collected bulk tank SCC. Somatic cell counts increased with an in- creasing intensity of

  4. Somatic cell count and milk yield in relation to haemoglobin concentration in Finnish dairy goats

    Microsoft Academic Search

    F. Atroshi; S. Sankari; U. B. Lindström

    1986-01-01

    Using 400 Finnish goats, the relationship between haemoglobin concentration and somatic cell counts and milk yield was studied. On the basis of haemoglobin concentration the goats were divided into two groups (> or ?1). Goats with high haemoglobin concentrations had a markedly higher milk yield and the milk had a lower somatic cell count. In contrast, goats with low haemoglobin

  5. Automated counting of cell bodies using Nissl stained cross-sectional images 

    E-print Network

    D'Souza, Aswin Cletus

    2009-05-15

    Cell count is an important metric in neurological research. The loss in numbers of certain cells like neurons has been found to accompany not only the deterioration of important brain functions but disorders like clinical ...

  6. Automated counting of cell bodies using Nissl stained cross-sectional images 

    E-print Network

    D'Souza, Aswin Cletus

    2008-10-10

    Cell count is an important metric in neurological research. The loss in numbers of certain cells like neurons has been found to accompany not only the deterioration of important brain functions but disorders like clinical ...

  7. For Most Children with HIV and Low Immune Cell Count, Cells Rebound After Treatment

    MedlinePLUS

    ... higher CD4+ counts.” — Rohan Hazra Chief, Maternal and Pediatric Infectious Disease Branch, NICHD Failure of CD4+ T cells , a ... author Rohan Hazra, chief of the Maternal and Pediatric Infectious Disease Branch at NIH’s Eunice Kennedy ShriverNational Institute of ...

  8. Effect of Thyroid Dysfunctions on Blood Cell Count and Red Blood Cell Indice

    PubMed Central

    Dorgalaleh, A; Mahmoodi, M; Varmaghani, B; Kiani node, F; Saeeidi Kia, O; Alizadeh, Sh; Tabibian, Sh; Bamedi, T; Momeni, M; Abbasian, S; Kashani Khatib, Z

    2013-01-01

    Background Thyroid hormones have a crucial role in metabolism and proliferation of blood cells. Thyroid dysfunction induces different effects on blood cells such as anemia, erythrocytosis leukopenia, thrombocytopenia, and in rare cases causes’ pancytopenia. It also alter RBC indices include MCV, MCH, MCHC and RDW. Thus this study attempted to evaluate effect of hypo & hyperthyroidism on blood cell count and RBC indices. Materials and Methods This study performed on 102 patients with hypothyroid (14.1 years), 84 with hyperthyroid (15.6 years) and 118 healthy individuals (15.2 years) as control group. Initially patients TSH level of patients was determined by ELISA method, and then according to TSH ranges (0.3-5.5µIU/mL) patients were divided into two Hyperthyroidism (TSH<0.3µIU/mL) and hypothyroidism (TSH>5.5µIU/mL) groups. Then, complete blood count was measured by cell counter. Finally, obtained results were analyzed by SPSS software. Results Analyzes of obtained data revealed statistically significant difference between two groups of patients in RBC count, MCH, MCHC, RDW, HB and HCT(P-value<0.05), but the difference was not significant for WBC and PLT counts and MCV (P-value>0.05). Conclusion In case of patients with unknown hematological dysfunctions, must be evaluated for thyroid hormones. PMID:24575274

  9. Counting White Blood Cells from a Blood Smear Using Fourier Ptychographic Microscopy

    PubMed Central

    Kulkarni, Rajan P.; Yang, Changhuei

    2015-01-01

    White blood cell (WBC) count is a valuable metric for assisting with diagnosis or prognosis of various diseases such as coronary heart disease, type 2 diabetes, or infection. Counting WBCs can be done either manually or automatically. Automatic methods are capable of counting a large number of cells to give a statistically more accurate reading of the WBC count of a sample, but the specialized equipment tends to be expensive. Manual methods are inexpensive since they only involve a conventional light microscope setup. However, it is more laborious and error-prone because the small field-of-view (FOV) of the microscope necessitates mechanical scanning of a specimen for counting an adequate number of WBCs. Here, we investigate the use of Fourier ptychographic microscopy (FPM) to bypass these issues of the manual methods. With a 2x objective, FPM can provide a FOV of 120 mm2 with enhanced resolution comparable to that of a 20x objective, which is adequate for non-differentially counting WBCs in just one FOV. A specialist was able to count the WBCs in FPM images with 100% accuracy compared to the count as determined from conventional microscope images. An automatic counting algorithm was also developed to identify WBCs from FPM’s captured images with 95% accuracy, paving the way for a cost-effective WBC counting setup with the advantages of both the automatic and manual counting methods. PMID:26186353

  10. Improving reliability of live/dead cell counting through automated image mosaicing.

    PubMed

    Piccinini, Filippo; Tesei, Anna; Paganelli, Giulia; Zoli, Wainer; Bevilacqua, Alessandro

    2014-12-01

    Cell counting is one of the basic needs of most biological experiments. Numerous methods and systems have been studied to improve the reliability of counting. However, at present, manual cell counting performed with a hemocytometer still represents the gold standard, despite several problems limiting reproducibility and repeatability of the counts and, at the end, jeopardizing their reliability in general. We present our own approach based on image processing techniques to improve counting reliability. It works in two stages: first building a high-resolution image of the hemocytometer's grid, then counting the live and dead cells by tagging the image with flags of different colours. In particular, we introduce GridMos (http://sourceforge.net/p/gridmos), a fully-automated mosaicing method to obtain a mosaic representing the whole hemocytometer's grid. In addition to offering more significant statistics, the mosaic "freezes" the culture status, thus permitting analysis by more than one operator. Finally, the mosaic achieved can thus be tagged by using an image editor, thus markedly improving counting reliability. The experiments performed confirm the improvements brought about by the proposed counting approach in terms of both reproducibility and repeatability, also suggesting the use of a mosaic of an entire hemocytometer's grid, then labelled trough an image editor, as the best likely candidate for the new gold standard method in cell counting. PMID:25438936

  11. Photoreceptor cell counting in adaptive optics retinal images using content-adaptive filtering

    NASA Astrophysics Data System (ADS)

    Mohammad, Fatimah; Ansari, Rashid; Wanek, Justin; Shahidi, Mahnaz

    2010-03-01

    Automated counting of photoreceptor cells in high-resolution retinal images generated by adaptive optics (AO) imaging systems is important due to its potential for screening and diagnosis of diseases that affect human vision. A drawback in recently reported photoreceptor cell counting methods is that they require user input of cell structure parameters. This paper introduces a method that overcomes this shortcoming by using content-adaptive filtering (CAF). In this method, image frequency content is initially analyzed to design a customized filter with a passband to emphasize cell structures suitable for subsequent processing. The McClellan transform is used to design a bandpass filter with a circularly symmetric frequency response since retinal cells have no preferred orientation. The automated filter design eliminates the need for manual determination of cell structure parameters, such as cell spacing. Following the preprocessing step, cell counting is performed on the binarized filtered image by finding regional points of high intensity. Photoreceptor cell count estimates using this automated procedure were found to be comparable to manual counts (gold standard). The new counting method when applied to test images showed overall improved performance compared with previously reported methods requiring user-supplied input. The performance of the method was also examined with retinal images with variable cell spacing.

  12. Effect of freezing on Fossomatic cell counting in ewe milk.

    PubMed

    Martínez, J R; Gonzalo, C; Carriedo, J A; San Primitivo, F

    2003-08-01

    Using the Fossomatic method, a total of 10,072 analytical somatic cell count (SCC) observations were carried out on 4760 aliquots taken from 70 individual ewe milk samples with the objective of studying whether freezing showed significant differences of SCC in comparison with refrigeration, according to different analytical conditions. These conditions were four preservation procedures (without preservation, potassium dichromate, azidiol, and bronopol), two storage temperatures (refrigeration and freezing), five milk ages within storage (24 h postcollection in refrigeration, and 24 h, 15, 30, and 60 d postcollection in freezing), two thawing types (rapid and slow), and two analytical temperatures (40 and 60 degrees C). Preservation, storage, and analytical temperature, type of thawing and milk age within storage, and most of the interactions showed a significant effect on the SCC variation. On average, the SCC was lower after freezing than in refrigeration. This effect depended specifically on type of preservation and analytical temperature of milk. The SCC of milk unpreserved or preserved with bronopol or potassium dichromate, and analyzed at 40 degrees C, was not affected by freezing; however, use of azidiol as a preservative before freezing, and heating the milk to 60 degrees C following thawing resulted in significantly decreased SCC. Milk age had little quantitative influence on SCC of thawed milk. The type of thawing (rapid and slow) did not significantly influence SCC of milk analyzed at 40 degrees C. As a result, when using properly handled samples, the Fossomatic method could be used to enumerate SCC in samples frozen over the 60 d. PMID:12939082

  13. Current automated 3D cell detection methods are not a suitable replacement for manual stereologic cell counting.

    PubMed

    Schmitz, Christoph; Eastwood, Brian S; Tappan, Susan J; Glaser, Jack R; Peterson, Daniel A; Hof, Patrick R

    2014-01-01

    Stereologic cell counting has had a major impact on the field of neuroscience. A major bottleneck in stereologic cell counting is that the user must manually decide whether or not each cell is counted according to three-dimensional (3D) stereologic counting rules by visual inspection within hundreds of microscopic fields-of-view per investigated brain or brain region. Reliance on visual inspection forces stereologic cell counting to be very labor-intensive and time-consuming, and is the main reason why biased, non-stereologic two-dimensional (2D) "cell counting" approaches have remained in widespread use. We present an evaluation of the performance of modern automated cell detection and segmentation algorithms as a potential alternative to the manual approach in stereologic cell counting. The image data used in this study were 3D microscopic images of thick brain tissue sections prepared with a variety of commonly used nuclear and cytoplasmic stains. The evaluation compared the numbers and locations of cells identified unambiguously and counted exhaustively by an expert observer with those found by three automated 3D cell detection algorithms: nuclei segmentation from the FARSIGHT toolkit, nuclei segmentation by 3D multiple level set methods, and the 3D object counter plug-in for ImageJ. Of these methods, FARSIGHT performed best, with true-positive detection rates between 38 and 99% and false-positive rates from 3.6 to 82%. The results demonstrate that the current automated methods suffer from lower detection rates and higher false-positive rates than are acceptable for obtaining valid estimates of cell numbers. Thus, at present, stereologic cell counting with manual decision for object inclusion according to unbiased stereologic counting rules remains the only adequate method for unbiased cell quantification in histologic tissue sections. PMID:24847213

  14. HIV viral load levels and CD4+ cell counts of youth in 14 cities

    PubMed Central

    Ellen, Jonathan M.; Kapogiannis, Bill; Fortenberry, J. Dennis; Xu, Jiahong; Willard, Nancy; Duval, Anna; Pace, Jill; Loeb, Jackie; Monte, Dina; Bethel, James

    2014-01-01

    Objectives To describe the HIV viral load and CD4+ cell counts of youth (12–24 years) in 14 cities from March 2010 through November 2011. Methods Baseline HIV viral load and CD4+ cell count data were electronically abstracted in a central location and in an anonymous manner through a random computer-generated coding system without any ability to link codes to individual cases. Results Among 1409 HIV reported cases, 852 participants had data on both viral load and CD4+ cell counts. Of these youth, 34% had CD4+ cell counts of 350 or less, 27% had cell counts from 351 to 500, and 39% had CD4+ cell counts greater than 500. Youth whose transmission risk was male-to-male sexual contact had higher viral loads compared with youth whose transmission risk was perinatal or heterosexual contact. Greater than 30% of those who reported male-to-male sexual contact had viral loads greater than 50 000 copies, whereas less than 20% of heterosexual contact youth had viral loads greater than 50 000 copies. There were no differences noted in viral load by type of testing site. Conclusion Most HIV-infected youth have CD4+ cell counts and viral load levels associated with high rates of sexual transmission. Untreated, these youth may directly contribute to high rates of ongoing transmission. It is essential that any public health test and treat strategy place a strong emphasis on youth, particularly young MSM. PMID:25028912

  15. The Geophysical Fluid Flow Cell Experiment

    NASA Technical Reports Server (NTRS)

    Hart, J. E.; Ohlsen, D.; Kittleman, S.; Borhani, N.; Leslie, F.; Miller, T.

    1999-01-01

    The Geophysical Fluid Flow Cell (GFFC) experiment performed visualizations of thermal convection in a rotating differentially heated spherical shell of fluid. In these experiments dielectric polarization forces are used to generate a radially directed buoyancy force. This enables the laboratory simulation of a number of geophysically and astrophysically important situations in which sphericity and rotation both impose strong constraints on global scale fluid motions. During USML-2 a large set of experiments with spherically symmetric heating were carried out. These enabled the determination of critical points for the transition to various forms of nonaxisymmetric convection and, for highly turbulent flows, the transition latitudes separating the different modes of motion. This paper presents a first analysis of these experiments as well as data on the general performance of the instrument during the USML-2 flight.

  16. Risk Factors Associated with Clinical Mastitis in Low Somatic Cell Count British Dairy Herds

    Microsoft Academic Search

    E. J. Peeler; M. J. Green; J. L. Fitzpatrick; K. L. Morgan; L. E. Green

    2000-01-01

    A cross-sectional survey of dairy farms with low bulk milk somatic cell counts was carried out to assess the level of clinical mastitis and to quantify risk factors associated with the incidence rate of clinical mastitis. Questionnaires were sent to 3009 milk operations with an annual mean bulk milk somatic cell count of less than 100,000 cells\\/ml during 1997. A

  17. Glial origin of rapidly adhering amniotic fluid cells

    Microsoft Academic Search

    P Aula; H von Koskull; K Teramo; O Karjalainen; I Virtanen; V P Lehto; D Dahl

    1980-01-01

    Rapidly adhering cells (RA cells) from the amniotic fluid of a pregnancy with fetal anencephaly were investigated by immunofluorescence assay with an antiserum against glial cells. After 24 hours' cultivation a high proportion of the cells showed positive glial-specific fluorescence, whereas no staining was seen in cells from samples of normal amniotic fluid. At the 24th week the mother was

  18. Seasonal variation of bulk milk somatic cell counts in UK dairy herds: Investigations of the summer rise

    Microsoft Academic Search

    M. J. Green; A. J. Bradley; H. Newton; W. J. Browne

    2006-01-01

    Individual cow somatic cell count (SCC) patterns were explored over a one year period in 33 dairy herds to investigate the reason for a summer rise in bulk milk somatic cell counts (BMSCC). Cow test day somatic cell counts were categorised according to the magnitude of change since the previous test day reading, to examine which categories were responsible for

  19. Predicting somatic cell count standard violations based on herd's bulk tank somatic cell count. Part II: Consistency index.

    PubMed

    Lukas, J M; Reneau, J K; Munoz-Zanzi, C; Kinsel, M L

    2008-01-01

    The present study examines the capability of 1,501 herds in the Upper Midwest and the performance of statistical process control charts and indices as a way of monitoring and controlling milk quality on the farm. For 24 mo, daily or every other day bulk tank somatic cell count (SCC) data were collected. Consistency indices for 5 different SCC standards were developed. The indices calculate the maximum variation allowed to meet a desired SCC level at a given mean bulk tank SCC and were used to identify herds not capable of meeting a specific SCC standard. Consistency index method was compared with a test identifying future bulk tank SCC standard violators based on herds' past violations. The performance of the consistency index test and the past violation method was evaluated by logistic regression. The comparison focused on detection probability and certainty associated with a result. For the 5 SCC levels, detection probability and certainty associated with a result ranged from 51 to 98%. Detection probability of all violators and certainty associated with a negative result was greater for the consistency index across all 5 SCC levels (by 0.7 to 7.4% and 2.1 to 5.1%, respectively). Control charts were plotted and monthly consistency indices calculated for individual farms. Charts in combination with the consistency indices would warn from 66 to 80% of the herds about an upcoming violation within 30 d before it occurred. They offer a proactive approach to maintaining consistently high milk quality. By assessing process capability and distinguishing between significant changes and random variation in bulk tank SCC, tools presented in this article encourage fact-based decisions in dairy farm milk quality management. PMID:18096968

  20. Non-parametric and integrated framework for segmenting and counting neuroblastic cells within neuroblastoma tumor images.

    PubMed

    Tafavogh, Siamak; Navarro, Karla Felix; Catchpoole, Daniel R; Kennedy, Paul J

    2013-06-01

    Neuroblastoma is a malignant tumor and a cancer in childhood that derives from the neural crest. The number of neuroblastic cells within the tumor provides significant prognostic information for pathologists. An enormous number of neuroblastic cells makes the process of counting tedious and error-prone. We propose a user interaction-independent framework that segments cellular regions, splits the overlapping cells and counts the total number of single neuroblastic cells. Our novel segmentation algorithm regards an image as a feature space constructed by joint spatial-intensity features of color pixels. It clusters the pixels within the feature space using mean-shift and then partitions the image into multiple tiles. We propose a novel color analysis approach to select the tiles with similar intensity to the cellular regions. The selected tiles contain a mixture of single and overlapping cells. We therefore also propose a cell counting method to analyse morphology of the cells and discriminate between overlapping and single cells. Ultimately, we apply watershed to split overlapping cells. The results have been evaluated by a pathologist. Our segmentation algorithm was compared against adaptive thresholding. Our cell counting algorithm was compared with two state of the art algorithms. The overall cell counting accuracy of the system is 87.65 %. PMID:23359256

  1. Optimization of a Cell Counting Algorithm for Mobile Point-of-Care Testing Platforms

    PubMed Central

    Ahn, DaeHan; Kim, Nam Sung; Moon, SangJun; Park, Taejoon; Son, Sang Hyuk

    2014-01-01

    In a point-of-care (POC) setting, it is critically important to reliably count the number of specific cells in a blood sample. Software-based cell counting, which is far faster than manual counting, while much cheaper than hardware-based counting, has emerged as an attractive solution potentially applicable to mobile POC testing. However, the existing software-based algorithm based on the normalized cross-correlation (NCC) method is too time- and, thus, energy-consuming to be deployed for battery-powered mobile POC testing platforms. In this paper, we identify inefficiencies in the NCC-based algorithm and propose two synergistic optimization techniques that can considerably reduce the runtime and, thus, energy consumption of the original algorithm with negligible impact on counting accuracy. We demonstrate that an Android™ smart phone running the optimized algorithm consumes 11.5× less runtime than the original algorithm. PMID:25195851

  2. CD8+ T-cells count in acute myocardial infarction in HIV disease in a predominantly male cohort.

    PubMed

    Badejo, Oluwatosin A; Chang, Chung-Chou; So-Armah, Kaku A; Tracy, Russell P; Baker, Jason V; Rimland, David; Butt, Adeel A; Gordon, Adam J; Rinaldo, Charles R; Kraemer, Kevin; Samet, Jeffrey H; Tindle, Hilary A; Goetz, Matthew B; Rodriguez-Barradas, Maria C; Bedimo, Roger; Gibert, Cynthia L; Leaf, David A; Kuller, Lewis H; Deeks, Steven G; Justice, Amy C; Freiberg, Matthew S

    2015-01-01

    Human Immunodeficiency Virus- (HIV-) infected persons have a higher risk for acute myocardial infarction (AMI) than HIV-uninfected persons. Earlier studies suggest that HIV viral load, CD4+ T-cell count, and antiretroviral therapy are associated with cardiovascular disease (CVD) risk. Whether CD8+ T-cell count is associated with CVD risk is not clear. We investigated the association between CD8+ T-cell count and incident AMI in a cohort of 73,398 people (of which 97.3% were men) enrolled in the U.S. Veterans Aging Cohort Study-Virtual Cohort (VACS-VC). Compared to uninfected people, HIV-infected people with high baseline CD8+ T-cell counts (>1065?cells/mm3) had increased AMI risk (adjusted HR=1.82, P<0.001, 95% CI: 1.46 to 2.28). There was evidence that the effect of CD8+ T-cell tertiles on AMI risk differed by CD4+ T-cell level: compared to uninfected people, HIV-infected people with CD4+ T-cell counts?200?cells/mm3 had increased AMI risk with high CD8+ T-cell count, while those with CD4+ T-cell counts<200?cells/mm3 had increased AMI risk with low CD8+ T-cell count. CD8+ T-cell counts may add additional AMI risk stratification information beyond that provided by CD4+ T-cell counts alone. PMID:25688354

  3. Effective Use of Dairy Herd Improvement Somatic Cell Counts in Mastitis Control

    Microsoft Academic Search

    Jeffrey K. Reneau

    1986-01-01

    The single most important factor affecting somatic cell count in milk is mammary gland infection status. In comparison, all other factors are minor. Consideration needs to be given to diurnal effects on Dairy Herd Improve- ment a.m.-p.m, sampling schemes. Somatic cell count linear score of 5 (283,000) appears to be a good choice of threshold for mastitis control applications. A

  4. Consequence for dairy herds in the United States of imposing different standards for somatic cell count

    Technology Transfer Automated Retrieval System (TEKTRAN)

    New European Union (E.U.) regulations may require that a somatic cell count (SCC) limit of 400,000 cells/mL for milk be met by every farm that contributes to pooled milk exported to Europe. In the United States, the standard is 750,000 cells/mL. Because bulk tank SCC is not readily available through...

  5. Somatic cell counts of milk from Dairy Herd Improvement herds during 2010

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Test-day data from all herds enrolled in Dairy Herd Improvement (DHI) somatic cell testing during 2010 were examined to assess the status of national milk quality. Somatic cell score (SCS) is reported to AIPL and was converted to somatic cell count (SCC) for calculating herd and State averages. The ...

  6. On distinguishing cause and consequence: Do high somatic cell counts lead to lower milk yield or does high milk yield lead to lower somatic cell count?

    Microsoft Academic Search

    L. E. Green; Y. H. Schukken; M. J. Green

    2006-01-01

    Researchers have reported that as milk yield increases composite milk somatic cell count (SCC) is diluted in cattle with no intramammary infection (IMI) and as a consequence, estimates of SCC from high yields are lower than estimates of SCC from low yields in dairy cows without an IMI. To date, estimates of reduced milk yield associated with high SCC because

  7. Differential white cell count and incident type 2 diabetes: the Insulin Resistance Atherosclerosis Study

    PubMed Central

    Lorenzo, Carlos; Hanley, Anthony J.; Haffner, Steven M.

    2014-01-01

    Aims/hypothesis White cell count has been shown to predict incident type 2 diabetes, but differential white cell count has received scant attention. We examined the risk of developing diabetes associated with differential white cell count and neutrophil:lymphocyte ratio and the effect of insulin sensitivity and subclinical inflammation on white cell associations. Methods Incident diabetes was ascertained in 866 participants aged 40–69 years in the Insulin Resistance Atherosclerosis Study after a 5 year follow-up period. The insulin sensitivity index (SI) was measured by the frequently sampled IVGTT. Results C-reactive protein was directly and independently associated with neutrophil (p<0.001) and monocyte counts (p<0.01) and neutrophil:lymphocyte ratio (p<0.001), whereas SI was inversely and independently related to lymphocyte count (p<0.05). There were 138 (15.9%) incident cases of diabetes. Demographically adjusted ORs for incident diabetes, comparing the top and bottom tertiles of white cell (1.80 [95% CI 1.10, 2.92]), neutrophil (1.67 [1.04, 2.71]) and lymphocyte counts (2.30 [1.41, 3.76]), were statistically significant. No association was demonstrated for monocyte count (1.18 [0.73, 1.90]) or neutrophil:lymphocyte ratio (0.89 [0.55, 1.45]). White cell and neutrophil associations were no longer significant after further adjusting for family history of diabetes, fasting glucose and smoking, but the OR comparing the top and bottom tertiles of lymphocyte count remained significant (1.92 [1.12, 3.29]). This last relationship was better explained by SI rather than C-reactive protein. Conclusions/interpretation A lymphocyte association with incident diabetes, which was the strongest association among the major white cell types, was partially explained by insulin sensitivity rather than subclinical inflammation. PMID:24141640

  8. Chloride and fluid secretion by cultured human polycystic kidney cells

    Microsoft Academic Search

    Darren P Wallace; Jared J Grantham; Lawrence P Sullivan

    1996-01-01

    Chloride and fluid secretion by cultured human polycystic kidney cells. Epithelial cells cultured from the renal cysts of patients with autosomal dominant polycystic kidney disease (ADPKD) secrete fluid via a process stimulated by adenosine 3?,5?-cyclic monophosphate (cAMP). We have investigated the hypothesis that fluid secretion by these cells is dependent on cAMP-mediated chloride secretion. Individual cultured ADPKD cells were suspended

  9. The Effects of Gamma and Proton Radiation Exposure on Hematopoietic Cell Counts in the Ferret Model.

    PubMed

    Sanzari, Jenine K; Wan, X Steven; Krigsfeld, Gabriel S; Wroe, Andrew J; Gridley, Daila S; Kennedy, Ann R

    2013-10-01

    Exposure to total-body radiation induces hematological changes, which can detriment one's immune response to wounds and infection. Here, the decreases in blood cell counts after acute radiation doses of ?-ray or proton radiation exposure, at the doses and dose-rates expected during a solar particle event (SPE), are reported in the ferret model system. Following the exposure to ?-ray or proton radiation, the ferret peripheral total white blood cell (WBC) and lymphocyte counts decreased whereas neutrophil count increased within 3 hours. At 48 hours after irradiation, the WBC, neutrophil, and lymphocyte counts decreased in a dose-dependent manner but were not significantly affected by the radiation type (?-rays verses protons) or dose rate (0.5 Gy/minute verses 0.5 Gy/hour). The loss of these blood cells could accompany and contribute to the physiological symptoms of the acute radiation syndrome (ARS). PMID:25356435

  10. ICSH guidelines for the verification and performance of automated cell counters for body fluids.

    PubMed

    Bourner, G; De la Salle, B; George, T; Tabe, Y; Baum, H; Culp, N; Keng, T B

    2014-12-01

    One of the many challenges facing laboratories is the verification of their automated Complete Blood Count cell counters for the enumeration of body fluids. These analyzers offer improved accuracy, precision, and efficiency in performing the enumeration of cells compared with manual methods. A patterns of practice survey was distributed to laboratories that participate in proficiency testing in Ontario, Canada, the United States, the United Kingdom, and Japan to determine the number of laboratories that are testing body fluids on automated analyzers and the performance specifications that were performed. Based on the results of this questionnaire, an International Working Group for the Verification and Performance of Automated Cell Counters for Body Fluids was formed by the International Council for Standardization in Hematology (ICSH) to prepare a set of guidelines to help laboratories plan and execute the verification of their automated cell counters to provide accurate and reliable results for automated body fluid counts. These guidelines were discussed at the ICSH General Assemblies and reviewed by an international panel of experts to achieve further consensus. PMID:24628711

  11. Automatic counting of FISH spots in interphase cells for prenatal characterization of aneuploidies

    NASA Astrophysics Data System (ADS)

    Ravkin, Ilya; Temov, Vladimir

    1999-06-01

    Fluorescent In-Situ Hybridization (FISH) is becoming an accepted technique for identification of aneuploidies in interphase fetal cells obtained by either CVS (chorionic villus sampling) or amniocentesis. Currently the analysis is done manually by a skilled operator and is a lengthy and fatiguing process. Applied Imaging is developing an automated procedure for counting FISH spots in these samples. Spot counting involves slide preparation, probe hybridization, filter selection, FISH image acquisition, image analysis, operator verification, and analysis of count distributions. We concentrate on the tasks starting with image acquisition. The following topics are covered: selection of appropriate cells, acquisition and processing of Z-stacks of FISH images for presentation and spot counting, background removal, formation of segmentation tree and selection of spot markers, growing of spot markers by means of constrained watershed, detection of irregular spots and flagging them for the user, time and accuracy compared with manual method, and applicability to a clinical research setting.

  12. Elevated White Blood Cell Count and Carotid Plaque Thickness The Northern Manhattan Stroke Study

    Microsoft Academic Search

    Mitchell S. Elkind; Jianfeng Cheng; Bernadette Boden-Albala; Myunghee C. Paik; Ralph L. Sacco

    Background and Purpose—Elevated leukocyte count has been associated with cardiovascular and cerebrovascular disease in several epidemiological studies. We sought to determine whether white blood cell count (WBC) is associated with carotid plaque thickness in a stroke-free, multiethnic cohort. Methods—For this cross-sectional analysis, WBC was measured in stroke-free community subjects undergoing carotid duplex Doppler ultrasound. Maximal internal carotid plaque thickness (MICPT)

  13. Consequence of changing standards for somatic cell count on US Dairy Herd Improvement herds

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Consequence of noncompliance with European Union (EU) and current US standards for somatic cell count (SCC) as well as SCC standards proposed by the National Milk Producers Federation was examined for US herds. Somatic cell scores (SCS) from 14,854 Dairy Herd Improvement (DHI) herds were analyzed. H...

  14. Predicting Progression in Glaucoma Suspects With Longitudinal Estimates of Retinal Ganglion Cell Counts

    PubMed Central

    Meira-Freitas, Daniel; Lisboa, Renato; Tatham, Andrew; Zangwill, Linda M.; Weinreb, Robert N.; Girkin, Christopher A.; Liebmann, Jeffrey M.; Medeiros, Felipe A.

    2013-01-01

    Purpose. We evaluated the ability of baseline and longitudinal estimates of retinal ganglion cell (RGC) counts in predicting progression in eyes suspected of having glaucoma. Methods. The study included 288 glaucoma suspect eyes of 288 patients followed for an average of 3.8 ± 1.0 years. Participants had normal standard automated perimetry (SAP) at baseline. Retinal nerve fiber layer thickness assessment was performed with optical coherence tomography (OCT). Progression was defined as development of repeatable abnormal SAP or glaucomatous progressive optic disc changes. Estimates of RGC counts were obtained by combining data from SAP and OCT according to a previously described method. Joint longitudinal survival models were used to evaluate the ability of baseline and rates of change in estimated RGC counts for predicting progression over time, adjusting for confounding variables. Results. A total of 48 eyes (17%) showed progression during follow-up. The mean rate of change in estimated RGC counts was ?18,987 cells/y in progressors versus ?8,808 cells/y for nonprogressors (P < 0.001). Baseline RGC counts and slopes of RGC loss were significantly predictive of progression, with HRs of 1.56 per 100,000 cells lower (95% confidence interval [CI], 1.18–2.08; P = 0.002) and 2.68 per 10,000 cells/y faster loss (95% CI, 1.22–5.90; P = 0.014), respectively. The longitudinal model including estimates of RGC counts performed significantly better than models including only structural or functional indexes separately. Conclusions. Baseline and longitudinal estimates of RGC counts may be helpful in predicting progression and performed significantly better than conventional approaches for risk stratification of glaucoma suspects. PMID:23661375

  15. Skin Tags: A Link Between Lesional Mast Cell Count/Tryptase Expression and Obesity and Dyslipidemia

    PubMed Central

    Salem, Samar Abdallah M; Attia, Enas AS; Osman, Wesam M; El Gendy, Marwa A

    2013-01-01

    Background: The etiology of skin tags (STs) is not fully understood. A relation to diabetes mellitus and obesity was suggested. Few studies of possible mast cells (MCs) involvement were reported. Tyrptase is a mast cell mediator and a potent fibroblast growth factor. It may provide a molecular link between mast cell activation and fibrosis. Aims: The aim was to assess clinical and laboratory findings in patients with STs, and the possible link between obesity, dyslipidemia, and lesional MC count/tryptase expression. Materials and Methods: A total of 20 patients with STs were subjected to clinical examination, estimation of body mass index (BMI), fasting blood glucose (FBG), postprandial blood glucose (PPBG), serum cholesterol and triglycerides, abdominal ultrasound for fatty liver assessment, in addition to study of MCs through staining for MC tryptase in two skin biopsies; lesional and nonlesional (control). Results: All patients showed abnormally high BMI and hypertriglyceridemia, with abnormal sonographic pattern in 15 patients (75%). STs number positively correlated with the age of patients. STs showed significantly higher MC counts and tryptase expression, compared with control skin (P < 0.001), with no correlation of the STs number or MC count with BMI, FBG, PPBG or serum cholesterol. Obese patients showed a significantly higher MC count than overweight and there was a positive correlation between MC count and serum triglycerides. Axilla and under breast STs showed a higher MC count compared with other sites. Conclusions: STs seem to be related to obesity and hypertriglyceridemia. MCs with their tryptase are possibly involved in pathogenesis of STs. MC count is related to the associated factors; obesity and serum triglycerides. MC tryptase expression is a reliable method for accurate tissue MC counting. PMID:23723485

  16. Reliability in somatic cell count measurement of clinical mastitis milk using DeLaval cell counter.

    PubMed

    Kawai, Kazuhiro; Hayashi, Tomohito; Kiku, Yoshio; Chiba, Tomoyuki; Nagahata, Hajime; Higuchi, Hidetoshi; Obayashi, Tetsu; Itoh, Seigo; Onda, Ken; Arai, Sachiko; Sato, Reiichiro; Oshida, Toshio

    2013-12-01

    Somatic cell counts (SCC) measurements are typically performed using quantitative methods, such as the Breed method (Breed) and the Fossomatic method (FSCC). The DeLaval cell counter (DCC) developed recently is a quantitative somatic cell counter with a low initial cost and superior portability. However, since the DCC was specifically developed for measuring SCC of ? 4 × 10(6) cells/mL milk from bulk tanks or individual cows, its reliability for estimating SCC that exceed this concentration has not yet been clarified. This study therefore examined whether it is possible to accurately measure SCC by diluting milk samples with initial SCC of 4 × 10(6) cells/mL, as seen in clinical mastitis milk. We collected milk samples from 99 quarters of 99 Holstein cows with clinical mastitis. These milk samples were diluted 10-fold with saline and thoroughly mixed before performing SCC measurement with the DCC. The correlation coefficients of SCC measured by the FSCC, Breed and DCC methods indicated strong correlations between each pair of methods. The findings showed that DCC can be used to identify bovine clinical mastitis milk and is useful as a quantitative SCC measurement device on farm sites. PMID:24128130

  17. Preoperative White Blood Cell Count and Risk of 30-Day Readmission after Cardiac Surgery

    PubMed Central

    Brown, Jeremiah R.; Landis, R. Clive; Chaisson, Kristine; Ross, Cathy S.; Dacey, Lawrence J.; Boss, Richard A.; Helm, Robert E.; Horton, Susan R.; Westbrook, Benjamin M.; LeBlond, Kelly; Quinn, Reed D.; Magnus, Patrick C.; Malenka, David J.; DiScipio, Anthony W.

    2013-01-01

    Approximately 1 in 5 patients undergoing cardiac surgery are readmitted within 30 days of discharge. Among the primary causes of readmission are infection and disease states susceptible to the inflammatory cascade, such as diabetes, chronic obstructive pulmonary disease, and gastrointestinal complications. Currently, it is not known if a patient's baseline inflammatory state measured by crude white blood cell (WBC) counts could predict 30-day readmission. We collected data from 2,176 consecutive patients who underwent cardiac surgery at seven hospitals. Patient readmission data was abstracted from each hospital. The independent association with preoperative WBC count was determined using logistic regression. There were 259 patients readmitted within 30 days, with a median time of readmission of 9 days (IQR 4–16). Patients with elevated WBC count at baseline (10,000–12,000 and >12,000?mm3) had higher 30-day readmission than those with lower levels of WBC count prior to surgery (15% and 18% compared to 10%–12%, P = 0.037). Adjusted odds ratios were 1.42 (0.86, 2.34) for WBC counts 10,000–12,000 and 1.81 (1.03, 3.17) for WBC count?>?12,000. We conclude that WBC count measured prior to cardiac surgery as a measure of the patient's inflammatory state could aid clinicians and continuity of care management teams in identifying patients at heightened risk of 30-day readmission after discharge from cardiac surgery. PMID:23970996

  18. Preoperative white blood cell count and risk of 30-day readmission after cardiac surgery.

    PubMed

    Brown, Jeremiah R; Landis, R Clive; Chaisson, Kristine; Ross, Cathy S; Dacey, Lawrence J; Boss, Richard A; Helm, Robert E; Horton, Susan R; Hofmaster, Patricia; Jones, Cheryl; Desaulniers, Helen; Westbrook, Benjamin M; Duquette, Dennis; Leblond, Kelly; Quinn, Reed D; Magnus, Patrick C; Malenka, David J; Discipio, Anthony W

    2013-01-01

    Approximately 1 in 5 patients undergoing cardiac surgery are readmitted within 30 days of discharge. Among the primary causes of readmission are infection and disease states susceptible to the inflammatory cascade, such as diabetes, chronic obstructive pulmonary disease, and gastrointestinal complications. Currently, it is not known if a patient's baseline inflammatory state measured by crude white blood cell (WBC) counts could predict 30-day readmission. We collected data from 2,176 consecutive patients who underwent cardiac surgery at seven hospitals. Patient readmission data was abstracted from each hospital. The independent association with preoperative WBC count was determined using logistic regression. There were 259 patients readmitted within 30 days, with a median time of readmission of 9 days (IQR 4-16). Patients with elevated WBC count at baseline (10,000-12,000 and >12,000?mm(3)) had higher 30-day readmission than those with lower levels of WBC count prior to surgery (15% and 18% compared to 10%-12%, P = 0.037). Adjusted odds ratios were 1.42 (0.86, 2.34) for WBC counts 10,000-12,000 and 1.81 (1.03, 3.17) for WBC count?>?12,000. We conclude that WBC count measured prior to cardiac surgery as a measure of the patient's inflammatory state could aid clinicians and continuity of care management teams in identifying patients at heightened risk of 30-day readmission after discharge from cardiac surgery. PMID:23970996

  19. Neutron confinement cell for investigating complex fluids

    SciTech Connect

    Kuhl, Tonya L.; Smith, Gregory S.; Israelachvili, Jacob N.; Majewski, Jaroslaw; Hamilton, William

    2001-03-01

    We describe an apparatus for measuring the molecular density and orientation of confined, ultrathin complex fluids under static and dynamic flow conditions. The device essentially couples the utility of the surface forces apparatus -- ability to control surface separation and alignment under applied loads -- with in situ structural characterization of the intervening material utilizing neutron reflectivity measurements. The apparatus is designed such that single crystal substrates of quartz or sapphire with areas up to tens of square centimeters can be kept parallel at controlled and well-defined separations from millimeters to less than 100 nm. The large substrate surface area enables direct structural measurements of the density profile of ''soft'' material placed between the aligned substrates. In addition, the cell is also designed to enable steady shear rates from 0.001 to 20 Hz to be applied in order to follow the dynamic structural response of the confined material, especially at the solid-solution interface. Faster shear rates of order 10{sup 4} can be obtained using oscillatory motion. Current design specifications focus on the use of neutron reflectivity to characterize the structure of end-grafted polymer brush layers, but the device can be employed to probe the structure of any complex fluid of interest and is amenable to other characterization techniques.

  20. White blood cell differential counts in severely leukopenic samples: a comparative analysis of different solutions available in modern laboratory hematology

    PubMed Central

    Kim, Ah Hyun; Lee, Wonbae; Kim, Myungshin; Kim, Yonggoo

    2014-01-01

    Background We evaluated the efficacy of white blood cell (WBC) differential counts in severely leukopenic samples by the Hematoflow method and by automated hematology analyzers and compared the results with manual counts. Methods EDTA-anticoagulated blood samples (175 samples) with WBC counts of 40-990/µL were selected. Hematoflow differential counts were performed in duplicates employing flow cytometry using the CytoDiff reagent and analysis software. Differential counts were also performed using the DxH 800 (Beckman Coulter) and XE-2100 (Sysmex) automated hematology analyzers. The sum of the manual counts by a hematology technician and a resident were used as the manual counts. Results The total analysis time and hands-on time required by the Hematoflow method were shorter than those required by manual counting. Hematoflow counts were reproducible, showed a good correlation with automated analyzers, and also showed strong correlation with manual counts (r > 0.8) in neutrophils, lymphocytes, and monocytes. None of the cases containing less than 4% blasts as analyzed by the Hematoflow method had blasts in the manual counts, but 8 cases of 21 cases (38.1%) with over 4% blasts by Hematoflow had blasts in manual counts. Conclusion Hematoflow counts of severely leukopenic samples were reproducible and showed a good correlation with manual counts in terms of neutrophil, lymphocyte, and monocyte counts. The Hematoflow method also detected the presence of blasts. Manual slide review is recommended when over 4% blasts are found by Hematoflow. PMID:25025014

  1. Genetic and Phenotypic Relationships Among Milk Yield and Somatic Cell Count Before and After Clinical Mastitis

    Microsoft Academic Search

    M. Koivula; E. A. Mäntysaari; E. Negussie; T. Serenius

    2005-01-01

    ABSTRACT This paper,studies,whether,cows,with,originally lower somatic,cell count (SCC) are more,susceptible,to clinical mastitis,(CM) than,cows,with,higher,somatic cell count, and evaluates the correlations between CM, SCC, and milk yield. Data were extracted from the Finnish national milk-recording,database,and from the health recording,system. First and second,lactation re- cords of 87,861 Ayrshire cows calving between January 1998 and December,2000 were included. Traits studied were incidence of CM, test-day

  2. Interphase argyrophilic nucleolar organiser regions and nucleolar counts in transitional cell bladder tumours

    PubMed Central

    Korneyev, I A; Mamaev, N N; Kozlov, V V; Rybakova, M G; Al-Shukri, S H

    2000-01-01

    Aims—To see whether a correlation exists between clinicopathological parameters, argyrophilic nucleolar organiser regions (AgNORs), and nucleolar counts in the nuclei of tumour cells in patients with transitional cell bladder carcinoma. Methods—Paraffin wax embedded sections from a total of 62 cases of primary transitional cell bladder carcinoma were stained with the silver colloid method. The numbers of individual silver grains (AgNORs) in nucleoli and the numbers of nucleoli were counted in 100 nuclei. The correlation between AgNORs and nucleolar counts and patients' sex, tumour grade, disease stage, recurrence pattern, and tumour related survival was analysed. Results—The numbers of nucleoli in tumour cells were higher in male patients (p < 0.032). AgNOR numbers correlated with tumour grade (p = 0.017) and recurrence (p = 0.046). In multivariate analysis, the variation coefficient of AgNOR scores was found to be the only independent predictor of the duration of tumour free period in patients with recurrent disease (p < 0.002). AgNOR scores and nucleolar counts were of no value in distinguishing superficial and invasive tumours or in predicting tumour related survival. Conclusions—AgNOR scores in transitional cell bladder carcinoma reflect variations in tumour biological behaviour; however, the clinical value of this technique in patients with urinary bladder carcinoma is limited. PMID:10897331

  3. Natural killer cell subsets in cerebrospinal fluid of patients with multiple sclerosis.

    PubMed

    Rodríguez-Martín, E; Picón, C; Costa-Frossard, L; Alenda, R; Sainz de la Maza, S; Roldán, E; Espiño, M; Villar, L M; Álvarez-Cermeño, J C

    2015-05-01

    Changes in blood natural killer (NK) cells, important players of the immune innate system, have been described in multiple sclerosis (MS). We studied percentages and total cell counts of different effector and regulatory NK cells in cerebrospinal fluid (CSF) of MS patients and other neurological diseases to gain clearer knowledge of the role of these cells in neuroinflammation. NK cell subsets were assessed by flow cytometry in CSF of 85 consecutive MS patients (33 with active disease and 52 with stable MS), 16 with other inflammatory diseases of the central nervous system (IND) and 17 with non-inflammatory neurological diseases (NIND). MS patients showed a decrease in percentages of different CSF NK subpopulations compared to the NIND group. However, absolute cell counts showed a significant increase of all NK subsets in MS and IND patients, revealing that the decrease in percentages does not reflect a real reduction of these immune cells. Remarkably, MS patients showed a significant increase of regulatory/effector (CD56(bright) /CD56(dim) ) NK ratio compared to IND and NIND groups. In addition, MS activity associated with an expansion of NK?T cells. These data show that NK cell subsets do not increase uniformly in all inflammatory neurological disease and suggest strongly that regulatory CD56(bright) and NK?T cells may arise in CSF of MS patients as an attempt to counteract the CNS immune activation characteristic of the disease. PMID:25565222

  4. Genotype by environment interaction for somatic cell score across bulk milk somatic cell count and days in milk

    Microsoft Academic Search

    M. P. L. Calus; L. L. G. Janss; R. F. Veerkamp

    2006-01-01

    The objective of this paper was to investigate the importance of a genotype x environment interaction (G x E) for somatic cell score (SCS) across levels of bulk milk somatic cell count (BMSCC), number of days in milk (DIM), and their interaction. Variance components were estimated with a model including random regressions for each sire on herd test-day BMSCC, DIM,

  5. An alternative staining method for counting red-eared slider turtle (Trachemys scripta) blood cells using crystal violet in cells diluted with 0.45% sodium chloride.

    PubMed

    Tsai, Chyong-Ying; Yu, Jane-Fang; Wang, Yu-Wen; Fan, Pei-Chia; Cheng, Ting-Yu; Wang, Lih-Chiann

    2014-09-01

    Various staining methods are available for reptilian species blood cell quantification. However, these methods have shown inaccurate differentiation limitations. The current study evaluates staining effects and blood cell counting results using an alternative method, counting blood cells diluted with 0.45% sodium chloride solution and stained with crystal violet. Blood samples from 8 red-eared slider turtles (Trachemys scripta) were collected. Red and white blood cell counts were performed using different methods: the unstained method, the Unopette method, Liu stain, and crystal violet method using blood cells diluted in various sodium chloride solution osmolarities. The staining properties and blood cell count results were compared. The crystal violet method using blood cells diluted in 0.45% sodium chloride solution delivered the best staining and counting results among all of the tested methods, with the lowest average coefficient of variance. The proposed method can easily be performed, serving as a feasible method for blood cell counting in chelonians. PMID:25080443

  6. Total and differential bulk cow milk somatic cell counts and their relation with antioxidant factors

    Microsoft Academic Search

    Houda Hamed; Abdelfettah El Feki; Ahmed Gargouri

    2008-01-01

    In the present study, the relationship between total bulk milk somatic cell counts (BMSCC), differential BMSCC (macrophage, lymphocyte, and polymorphonuclear leukocytes), and antioxidant enzymes was investigated. Forty-three samples of bulk milk were selected randomly from eight dairy farms in the region of Sfax (Tunisia) in winter, from November 2005 to February 2006. Bulk milk samples were analyzed for antioxidant enzymes

  7. Total and differential bulk cow milk somatic cell counts and their relation with lipolysis

    Microsoft Academic Search

    Ahmed Gargouri; Houda Hamed; Abdelfettah ElFeki

    2008-01-01

    In the present study, the relationship between total bulk milk somatic cell counts (BMSCC), differential BMSCC (macrophage, lymphocyte and polymorphonuclear leukocytes) and lipolysis level was investigated. Eighty samples of bulk milk were selected randomly from eight dairy farms in the region of Sfax (Tunisia) from November 2005 to April 2006. Bulk milk samples were analyzed for titratable acidity, total solids,

  8. Transcript counting in single cells reveals dynamics of rDNA transcription

    E-print Network

    van Oudenaarden, Alexander

    REPORT Transcript counting in single cells reveals dynamics of rDNA transcription Rui Zhen Tan1 and transcription; RNA Keywords: rDNA; single-molecule FISH; Rpd3 This is an open-access article distributed under copies of tandemly repeated rRNA genes. The regulation of rDNA transcription occurs by controlling both

  9. Quarter milk somatic cell count in infected dairy cows: a meta-analysis

    Microsoft Academic Search

    Belgacem Djabri; Nathalie Bareille; Henri Seegers

    2002-01-01

    The aim of this paper was to evaluate the effects associated with intramammary infection (IMI) by a bacterium or a group of bacteria (Staphylococcus aureus, Streptococcus agalactiae, Strep- tococcus dysgalactiae, Streptococcus uberis, coliforms, Staphylococci other than S. aureus ,a nd Corynebacterium bovis) on the somatic cell count (SCC) in quarter milk of dairy cows. Papers se- lected for analysis had

  10. Survey of Milking Hygiene Practices and Their Relationships to Somatic Cell Counts and Milk Production

    Microsoft Academic Search

    J. E. Moxley; B. W. Kennedy; B. R. Downey; J. S. T. Bowman

    1978-01-01

    A total of 581 official test herds on the Quebec Dairy Herd Analysis Service were surveyed to measure frequency of practices of milking hygiene and effects of milking hygiene on herd averages of somatic cell counts and milk production. Over 99% of the dairymen washed or sprayed cows' udders, 55% used a sepa- rate towel for washing, 18% dried udders

  11. Cost Benefit Analysis of Lactation Therapy with Somatic Cell Counts as Indications for Treatment1

    Microsoft Academic Search

    Michael P. McDermott; Hollis N. Erb; Roger P. Natzke; Frances D. Barnes; David Bray

    1983-01-01

    Lactating dairy cows (487) from five commercial herds were in a study of benefits from lactation therapy of sub- clinical mastitis. Bacterial isolations and composite milk samples for somatic cell counts were taken from each cow each month for 15 mo. Cows (254) in the ex- perimental group were infused with cephapirin in all quarters for two con- secutive milkings

  12. Somatic cell count as a selection criterion for mastitis resistance in dairy cattle

    Microsoft Academic Search

    J. Philipsson; G. Ral; B. Berglund

    1995-01-01

    The purpose of this study was to investigate whether it is possible to increase the resistance to clinical mastitis by selecting for bulls siring daughters whose milk has a low somatic cell count. Data from the animal disease recording scheme, including all veterinary treatments for clinical mastitis, were used in the analysis. A total of 1462 progeny tested Swedish Red

  13. Estimation of Variance Components for Somatic Cell Counts to Determine Thresholds for Uninfected Quarters

    Microsoft Academic Search

    A. J. Schepers; T. J. G. M. Lam; Y. H. Schukken; J. B. M. Wilmink; W. J. A. Hanekamp

    1997-01-01

    The objective of this study was to determine the factors affecting somatic cell count (SCC), to esti- mate variance components of these factors, and to calculate and evaluate the thresholds for intramam- mary infection based on SCC. The infection status from 22,467 quarter milk samples from 544 cows in seven herds was determined. Infection status was the most important factor

  14. The relationship between dairy cow hygiene and somatic cell count in milk

    Microsoft Academic Search

    A. C. Sant’Anna; M. J. R. Paranhos da Costa

    2011-01-01

    Corporal hygiene is an important indicator of welfare for dairy cows and is dependent on facilities, climate conditions, and the behavior of the animals. The objectives of this study were to describe how the hygiene conditions of dairy cows vary over time and to assess whether a relationship exists between hygiene and somatic cell count (SCC) in milk. Monthly hygiene

  15. Relationship of somatic cell counts in goat milk to mastitis and productivity

    Microsoft Academic Search

    George F. W Haenlein

    2002-01-01

    This review covers research since the Symposium at Bella, Italy, in 1994, and as published in three major dairy research journals. It examines in particular non-pathological influences on somatic cell counts (SCCs) levels as they are unique for goat milk and different from cow milk in order to aid towards progress for establishing equitable quality standards for goat and sheep

  16. Bulk tank milk somatic cell count and sources of raw milk contamination with mastitis pathogens

    Microsoft Academic Search

    D. Rysanek; V. Babak; M. Zouharova

    ABstr Act : The objective of this study was to probe the relationship between prevalence of selected principal mastitis pathogens and somatic cell counts in bulk tank milk samples. The sources of milk contamination were evaluated. The samples were collected from 298 dairy herds (with approximately 32 000 dairy cows). Only 48.3% of the bulk tank milk samples were free

  17. Mammary Pathogens and Their Relationship to Somatic Cell Count and Milk Yield Losses in Dairy Ewes

    Microsoft Academic Search

    C. Gonzalo; A. Ariznabarreta; J. A. Carriedo; F. San Primitivo

    2002-01-01

    A total of 9592 samples of half udder milk were col- lected monthly throughout lactation for bacteriological and somatic cell count (SCC) study from 1322 Churra ewe lactations from seven separate flocks enrolled in the recording scheme of the National Association of Spanish Churra Breeders in the Castile-Leon region of Spain. Statistical analyses were carried out from a mixed model

  18. Composition and bulk tank somatic cell counts of milk from dairy goat herds in Southeastern Brazil

    Microsoft Academic Search

    Maria Aparecida; Vasconcelos Paiva BRITO; Carla LANGE; Rafael Guedes FONSECA; Yuri de Almeida SILVA

    The milk composition and somatic cell count (SCC) are requirements for assessment milk quality and mastitis in goat herds. Studies conducted with dairy goat herds indicated that the milk composition differed among them to due to factors such as genetic, feeding, system of production, stage of lactation, year and year-season. The objective of this study was to assess SCC and

  19. Relationship of Somatic Cell Count with Milk Yield and Composition in Chinese Holstein Population

    Microsoft Academic Search

    Jia-zhong GUO; Xiao-lin LIU; A-juan XU; Zhi XIA

    2010-01-01

    The objective of this study was to analyze the relationship of somatic cell count (SCC) with milk yield, fat and protein percentage, fat and protein yield using analysis of variance and correlation analysis in Chinese Holstein population. The 10 524 test-day records of 568 Chinese Holstein Cattle were obtained from 2 commercial herds in Xi'an region of China during February

  20. In Vivo Imaging and Counting of Rat Retinal Ganglion Cells Using a Scanning Laser Ophthalmoscope

    Microsoft Academic Search

    Tomomi Higashide; Ichiro Kawaguchi; Shinji Ohkubo; Hisashi Takeda; Kazuhisa Sugiyama

    2006-01-01

    RESULTS. Fluorescent RGCs were visible in vivo with an SLO. RGC survival decreased gradually after the crush. In the retina after the optic nerve crush, newly emerged DiA fluorescence detected by image overlay analysis corresponded to fluores- cent cells morphologically different from RGCs in the retinal flatmount and was colocalized mostly with lectin-stained mi- croglial processes. RGC counts by SLO

  1. The future role of CD4 cell count for monitoring antiretroviral therapy.

    PubMed

    Ford, Nathan; Meintjes, Graeme; Pozniak, Anton; Bygrave, Helen; Hill, Andrew; Peter, Trevor; Davies, Mary-Ann; Grinsztejn, Beatriz; Calmy, Alexandra; Kumarasamy, N; Phanuphak, Praphan; deBeaudrap, Pierre; Vitoria, Marco; Doherty, Meg; Stevens, Wendy; Siberry, George K

    2015-02-01

    For more than two decades, CD4 cell count measurements have been central to understanding HIV disease progression, making important clinical decisions, and monitoring the response to antiretroviral therapy (ART). In well resourced settings, the monitoring of patients on ART has been supported by routine virological monitoring. Viral load monitoring was recommended by WHO in 2013 guidelines as the preferred way to monitor people on ART, and efforts are underway to scale up access in resource-limited settings. Recent studies suggest that in situations where viral load is available and patients are virologically suppressed, long-term CD4 monitoring adds little value and stopping CD4 monitoring will have major cost savings. CD4 cell counts will continue to play an important part in initial decisions around ART initiation and clinical management, particularly for patients presenting late to care, and for treatment monitoring where viral load monitoring is restricted. However, in settings where both CD4 cell counts and viral load testing are routinely available, countries should consider reducing the frequency of CD4 cell counts or not doing routine CD4 monitoring for patients who are stable on ART. PMID:25467647

  2. Analysis of Treatment for HIV-infected Patients Considering CD4 T Cell Count in STI

    Microsoft Academic Search

    Ki Yeon Park; Han Byul Chung; Chung Choo Chung

    2006-01-01

    Determining drug dosage for STI (structured treatment interruption) according to the clinically healthy CD4 T cell count has recently been proposed due to its simple scheduling and effectiveness. In this paper, we investigated its effectiveness and validity from a mathematical biological point of view and analyzed the human immune system based on a mathematical model of a HIV-infected patient. Compared

  3. Low somatic cell count : a risk factor for subsequent clinical mastitis in a dairy herd

    Microsoft Academic Search

    W. Suriyasathaporn; Y.H. Schukken; M. Nielen; A. Brand

    2000-01-01

    A case-control study was conducted to evaluate factors measured at the udder inflammation-free state as risk factors for subsequent clinical mastitis. The factors including somatic cell count (SCC), body condition score, milk yield, percentages of milk fat and milk protein, and diseases were evaluated for their association with the results of udder inflammatory response. The results of the response were

  4. Influence of Storage and Preservation on Fossomatic Cell Count and Composition of Goat Milk

    Microsoft Academic Search

    A. Sánchez; D. Sierra; C. Luengo; J. C. Corrales; C. T. Morales; A. Contreras; C. Gonzalo

    2005-01-01

    This study was designed to evaluate the effects of different test conditions on the somatic cell count (SCC) and composition of goat milk. To this end, 3600 tests were performed on 1800 aliquots taken from 40 goat milk samples using a combined instrument set-up based on flow cytometry for SCC and Fourier transform infra- red analysis for fat, total protein,

  5. Genetic analysis of somatic cell count and milk traits in Manchega ewes

    Microsoft Academic Search

    M. Serrano; M. D. Pérez-Guzmán; V. Montoro; J. J. Jurado

    2003-01-01

    A total of 36?873 lactation records of Somatic Cell Counts (SCC) and 32?844 of milk yield, protein and dry matter contents of 28?694 Spanish Manchega ewes from 118 herds were used to estimate genetic parameters under different approaches. In a first step estimations were carried out under multivariate animal models including lactation mean somatic cell score (SCSL) and other milk

  6. Antibiotic therapy for subclinical mastitis in early lactation; effects on infection, somatic cell count & milk production

    Microsoft Academic Search

    S. M. Mwakipesile; C. W. Holmes; Y. F. Moore

    1983-01-01

    Cows with consistently high somatic cell counts (SCC) and bacterial infection, but no clinical signs of mastitis, were identified early in lactation. Some of these quarters were treated with Cloxacillin, while other similar quarters were left untreated. Prior to treatment, 74% of infected quarters had SCC higher than 300,000 cells\\/ml while 85% of uninfected quarters has SCC lower than 300,000

  7. Correlations Between Somatic Cells Count and Milk Composition with Regard to the Season

    Microsoft Academic Search

    Marija RAJÈEVIÈ; Klemen POTOÈNIK

    SUMMARY On four farms with 1100 cows of Black-and-White breed the correlations between somatic cell count (SCC) in milk and milk composition in different seasons of the year were studied. Three-year data from AP milk recording were analysed (years 2000 to 2002), which meant 29,298 samples of milk. Milk fat, proteins and lactose were determined using Milco-Scan while somatic cells

  8. Evaluation of the Overall Accuracy of the DeLaval Cell Counter for Somatic Cell Counts in Ovine Milk

    Microsoft Academic Search

    C. Gonzalo; B. Linage; J. A. Carriedo; F. de la Fuente; F. San Primitivo

    2006-01-01

    The DeLaval cell counter (DCC) is a portable device designed for on-farm somatic cell count (SCC) analysis in bovine milk. This study evaluated the performance of the DCC when analyzing ovine milk. A total of 29 composite ovine milk samples, ranging between 20 × 103 and 2,200 × 103 cells\\/mL, were divided into 15 aliquots\\/ milk sample corresponding to 5

  9. Automated imaging, identification, and counting of similar cells from digital hologram reconstructions

    NASA Astrophysics Data System (ADS)

    Mihailescu, Mona; Scarlat, Mihaela; Gheorghiu, Alexandru; Costescu, Julia; Kusko, Mihai; Paun, Irina Alexandra; Scarlat, Eugen

    2011-07-01

    This paper presents our method, which simultaneously combines automatic imaging, identification, and counting with the acquisition of morphological information for at least 1000 blood cells from several three-dimensional images of the same sample. We started with seeking parameters to differentiate between red blood cells that are similar but different with respect to their development stage, i.e., mature or immature. We highlight that these cells have different diffractive patterns with complementary central intensity distribution in a given plane along the propagation axis. We use the Fresnel approximation to simulate propagation through cells modeled as spheroid-shaped phase objects and to find the cell property that has the dominant influence on this behavior. Starting with images obtained in the reconstruction step of the digital holographic microscopy technique, we developed a code for automated simultaneous individual cell image separation, identification, and counting, even when the cells are partially overlapped on a slide, and accurate measuring of their morphological features. To find the centroids of each cell, we propose a method based on analytical functions applied at threshold intervals. Our procedure separates the mature from the immature red blood cells and from the white blood cells through a decision based on gradient and radius values.

  10. HEMODOSE: A Biodosimetry Tool Based on Multi-type Blood Cell Counts

    PubMed Central

    Hu, Shaowen; Blakely, William F.; Cucinotta, Francis A.

    2015-01-01

    Abstract Peripheral blood cell counts are important biomarkers of radiation exposure. In this work, a simplified compartmental modeling approach is applied to simulate the perturbation of the hematopoiesis system in humans after radiation exposure, and HemoDose software is reported to estimate individuals’ absorbed doses based on multi-type blood cell counts. Testing with patient data in some historical accidents indicates that either single or serial granulocyte, lymphocyte, leukocyte, and platelet counts after exposure can be robust indicators of the absorbed doses. In addition, such correlation exists not only in the early time window (1 or 2 d) but also in the late phase (up to 4 wk) after exposure, when the four types of cell counts are combined for analysis. These demonstrate the capability of HemoDose as a rapid point-of-care diagnostic or centralized high-throughput assay system for personnel exposed to unintended high doses of radiation, especially in large-scale nuclear/radiological disaster scenarios involving mass casualties. PMID:26011498

  11. HEMODOSE: A Biodosimetry Tool Based on Multi-type Blood Cell Counts.

    PubMed

    Hu, Shaowen; Blakely, William F; Cucinotta, Francis A

    2015-07-01

    Peripheral blood cell counts are important biomarkers of radiation exposure. In this work, a simplified compartmental modeling approach is applied to simulate the perturbation of the hematopoiesis system in humans after radiation exposure, and HemoDose software is reported to estimate individuals' absorbed doses based on multi-type blood cell counts. Testing with patient data in some historical accidents indicates that either single or serial granulocyte, lymphocyte, leukocyte, and platelet counts after exposure can be robust indicators of the absorbed doses. In addition, such correlation exists not only in the early time window (1 or 2 d) but also in the late phase (up to 4 wk) after exposure, when the four types of cell counts are combined for analysis. These demonstrate the capability of HemoDose as a rapid point-of-care diagnostic or centralized high-throughput assay system for personnel exposed to unintended high doses of radiation, especially in large-scale nuclear/radiological disaster scenarios involving mass casualties. PMID:26011498

  12. Spectral counting assessment of protein dynamic range in cerebrospinal fluid following depletion with plasma-designed immunoaffinity columns

    PubMed Central

    2011-01-01

    Background In cerebrospinal fluid (CSF), which is a rich source of biomarkers for neurological diseases, identification of biomarkers requires methods that allow reproducible detection of low abundance proteins. It is therefore crucial to decrease dynamic range and improve assessment of protein abundance. Results We applied LC-MS/MS to compare the performance of two CSF enrichment techniques that immunodeplete either albumin alone (IgYHSA) or 14 high-abundance proteins (IgY14). In order to estimate dynamic range of proteins identified, we measured protein abundance with APEX spectral counting method. Both immunodepletion methods improved the number of low-abundance proteins detected (3-fold for IgYHSA, 4-fold for IgY14). The 10 most abundant proteins following immunodepletion accounted for 41% (IgY14) and 46% (IgYHSA) of CSF protein content, whereas they accounted for 64% in non-depleted samples, thus demonstrating significant enrichment of low-abundance proteins. Defined proteomics experiment metrics showed overall good reproducibility of the two immunodepletion methods and MS analysis. Moreover, offline peptide fractionation in IgYHSA sample allowed a 4-fold increase of proteins identified (520 vs. 131 without fractionation), without hindering reproducibility. Conclusions The novelty of this study was to show the advantages and drawbacks of these methods side-to-side. Taking into account the improved detection and potential loss of non-target proteins following extensive immunodepletion, it is concluded that both depletion methods combined with spectral counting may be of interest before further fractionation, when searching for CSF biomarkers. According to the reliable identification and quantitation obtained with APEX algorithm, it may be considered as a cheap and quick alternative to study sample proteomic content. PMID:21906361

  13. Eosinophil count - absolute

    MedlinePLUS

    ... you have certain allergic diseases, infections, and other medical conditions. ... to show up as orange-red granules. The technician then counts how ... white blood cell count to give the absolute eosinophil count.

  14. A Comparison of Pyronin Y-Methyl Green Stain and Methylene Blue Stain for Somatic Cell Count in Sheep Milk

    Microsoft Academic Search

    Emily Mirek; Stacey O’Donnell

    2007-01-01

    Somatic cell count is a key method used to evaluate the quality of milk in today’s growing dairy sheep industry. Somatic cells are body cells, primarily the white blood cells, found in a milk sample. If an infection such as mastitis is present, the number of somatic cells in the milk increases (Gonzalo, et al. 1992). Producers routinely perform somatic

  15. Low Counts of Plasmacytoid Dendritic Cells after Engraftment Are Associated with High Early Mortality after Allogeneic Stem Cell Transplantation.

    PubMed

    Gonçalves, Matheus Vescovi; Yamamoto, Mihoko; Kimura, Eliza Yurico Sugano; Colturato, Vergílio Antônio Rensi; de Souza, Mair Pedro; Mauad, Marcos; Ikoma, Maura Valerio; Novis, Yana; Rocha, Vanderson; Ginani, Valeria Cortez; Wanderley de Oliveira Felix, Olga Margareth; Seber, Adriana; Kerbauy, Fabio Rodrigues; Hamerschlak, Nelson; Orfao, Alberto; Rodrigues, Celso Arrais

    2015-07-01

    Dendritic cells (DCs) are antigen-presenting cells that drive immune responses and tolerance and are divided in different subsets: myeloid DCs (mDCs: lineage-; HLA-DR+, 11c+), plasmacytoid dendritic cells (pDCs: HLA-DR+, CD123+), and monocyte-derived DCs (moDC: lineage-, 11c+, 16+). After hematopoietic stem cell transplantation (HSCT), low DC counts in the recipients' peripheral blood (PB) have been associated with worse outcomes, but the relevance of DC graft content remains unclear, and there are few data in the setting of unrelated donor HSCT. We evaluated the DC graft content and monitored DC recovery in PB from 111 HSCT recipients (median age, 17 years; range 1 to 74), who received bone marrow (46%), umbilical cord blood (32%), or PB (22%) from unrelated (81%) or related donors (19%). In 86 patients with sustained allogeneic recovery, patients with higher counts of all DC subsets (pDC, mDC, and moDC) 3 weeks after engraftment had lower incidence of nonrelapse mortality (NMR) and acute graft-versus-host disease (aGVHD) and better survival. pDC counts were associated with more striking results: patients with higher pDC counts had much lower incidences of NRM (3% versus 47%, P < .0001), lower incidence of aGVHD (24% versus 67%, P < .0001), and better overall survival (92% versus 45%, P < .0001). In contrast, higher pDC counts in the graft was associated with an increased risk of aGVHD (55% versus 26%, P = .02). Our results indicate that DC counts are closely correlated with HSCT outcomes and warrant further prospective evaluation and possible early therapeutic interventions to ameliorate severe aGVHD and decrease mortality. PMID:25792371

  16. Short communication: factors influencing time to CD4(+) T cell counts >200 cells/mm(3) in HIV-infected individuals with CD4(+) T cell <50 cells/mm(3) at the time of starting combination antiretroviral therapy.

    PubMed

    Ku, Nam Su; Song, Young Goo; Han, Sang Hoon; Kim, Sun Bean; Kim, Hye-won; Jeong, Su Jin; Kim, Chang Oh; Kim, June Myung; Choi, Jun Yong

    2012-12-01

    We evaluated factors influencing time to CD4(+) T cell counts >200 cells/mm(3) in HIV-infected individuals with CD4(+) T cell <50 cells/mm(3) starting combination antiretroviral therapy (cART). We included a total of 29 patients on successful cART for more than 1 year. In a logistic regression model, higher pre-cART CD4(+) T cell counts were significantly associated with shorter time to CD4(+) T cell counts >200cells/mm(3) in HIV-infected individuals with baseline CD4(+) T cell <50 cells/mm(3). In survival analysis, patients having higher pre-cART CD4(+) T cell counts, especially 40-49 cells/mm(3), were at significantly higher risk of achieving CD4(+) T cell counts >200 cells/mm(3). PMID:22632127

  17. Signet ring cells in ascitic fluid -- a diagnostic dilemma.

    PubMed

    Padma, K; Sundari, N

    2011-07-01

    Traditional teaching is that signet ring cells are the hall-mark of ovarian metastatic tumours. Paracentesis done for therapeutic purpose showed these cells and momentarily misled us. Other differentiating data are important in the correct management of cases with signet ring cells found in either ascitic fluid or on histopathologic examination. PMID:22315843

  18. A comparative study of white blood cell counts and disease risk in carnivores.

    PubMed Central

    Nunn, Charles L; Gittleman, John L; Antonovics, Janis

    2003-01-01

    In primates, baseline levels of white blood cell (WBC) counts are related to mating promiscuity. It was hypothesized that differences in the primate immune system reflect pathogen risks from sexually transmitted diseases (STDs). Here, we test for the generality of this result by examining hypotheses involving behavioural, ecological and life-history factors in carnivores. Again, we find a significant correlation in carnivores between mating promiscuity and elevated levels of WBC counts. In addition, we find relationships with measures of sociality, substrate use and life-history parameters. These comparative results across independent taxonomic orders indicate that the evolution of the immune system, as represented by phylogenetic differences in basal levels of blood cell counts, is closely linked to disease risk involved with promiscuous mating and associated variables. We found only limited support for an association between the percentage of meat in the diet and WBC counts, which is consistent with the behavioural and physiological mechanisms that carnivores use to avoid parasite transmission from their prey. We discuss additional comparative questions related to taxonomic differences in disease risk, modes of parasite transmission and implications for conservation biology. PMID:12639313

  19. Environmental Factors Influencing Test-Day Somatic Cell Counts in Holsteins

    Microsoft Academic Search

    B. W. Kennedy; M. S. Sethar; A. K. W. Tong; J. E. Moxley; B. R. Downey

    1982-01-01

    ABSTRACT Between February and December, 1977, 133,493 test-day observations of somatic cell count were taken on 27,009 Holstein,cows,in,676 herds,on,the Quebec Dairy Herd Analysis Service. Data were transformed to a log (natural) scale, and,analyses,were,separate,within,lacta- tion age group (42, 3, 4, 5, and \\/>6 yr). Joint estimates,of fixed effects of month of test, age of sample at time of labora- tory analysis,

  20. Variation in blood leucocytes, somatic cell count, yield and composition of milk of crossbred goats

    Microsoft Academic Search

    Mainak Das; Mahendra Singh

    2000-01-01

    Ten multiparous crossbred goats, five each of alpine×beetal (AB) and saanen×beetal (SB) were selected from the National Dairy Research Institute goat herd immediately after parturition. These were managed as per the practices followed in the institute’s goatherd. Blood and milk samples were collected at biweekly intervals from day 14 post-kidding for 22 weeks (154 days). Somatic cell count, electrical conductivity,

  1. Genetic and environmental correlations between test-day somatic cell count and milk yield traits

    Microsoft Academic Search

    M Haile-Mariam; P. J Bowman; M. E Goddard

    2001-01-01

    Australian Holstein cattle data were analysed to estimate genetic and environmental correlations between test-day yield (milk, protein, fat) and somatic cell count (SCC) using a random regression sire model. The estimated correlations were also compared to those from multi-trait and repeatability models. Estimates obtained from different models showed a similar trend in which genetic correlations between yield and logSCC were

  2. Effect of Mastitis Treatment and Somatic Cell Counts on Milk Yield in Danish Organic Dairy Cows

    Microsoft Academic Search

    T. W. Bennedsgaard; C. Enevoldsen; S. M. Thamsborg; M. Vaarst

    2003-01-01

    Production and disease data from 17,488 lactations in 48 Danish organic dairy herds from 1997 to 2001 were analyzed to obtain estimates on the effect of so- matic cell counts (SCC) and mastitis treatment on milk production. A multilevel three-parameter piecewise random coefficients linear model with energy-corrected milk (ECM) as dependent variable and herd, lactation, and test days as levels,

  3. Treatment of childhood lymphocytic leukaemia with high white-cell counts

    Microsoft Academic Search

    H Ekert; K D Waters; P J Smith; R N Matthews; W M Ellis

    1978-01-01

    Combination chemotherapy with cytosine arabinoside, cyclophosphamide and L-asparaginase (Asnase) was given to 22 children with acute lymphocytic leukaemia (ALL) with a white-cell count greater than 30 X 10(9)\\/1, and other features suggestive of poor prognosis. Complete remission was induced in all patients--in 19 after 2 courses of chemotherapy and in the remainder after a third course. During induction, neutropenia occurred

  4. Obesity and the White Blood Cell Count: Changes with Sustained Weight Loss

    Microsoft Academic Search

    John B. Dixon; Paul E O' Brien

    2006-01-01

    Background: Obesity is a chronic inflammatory condition, and elevated white blood cell counts (WBC) have widely recognized\\u000a associations with inflammatory conditions. The authors explored the relationship between the WBC and degree of obesity, basic\\u000a anthropometry, and clinical and biochemical markers of the metabolic syndrome at baseline, and with weight loss following\\u000a Lap-Band surgery. Methods: 477 patients with complete biochemical and

  5. Relationship among specific bacterial counts and total bacterial and somatic cell counts and factors influencing their variation in ovine bulk tank milk.

    PubMed

    de Garnica, M L; Linage, B; Carriedo, J A; De La Fuente, L F; García-Jimeno, M C; Santos, J A; Gonzalo, C

    2013-02-01

    To analyze the relationship among the counts of different organisms and total bacterial count (BTTBC) and somatic cell count (BTSCC) as determined in dairy laboratories in ovine bulk tank milk, 751 bulk tank milk samples from 205 dairy sheep flocks belonging to Consortium for Ovine Promotion (CPO) were collected between January and December 2011. Four samplings were carried out in each flock, once per season, throughout 1 yr. Variables analyzed were bulk tank counts of thermoduric, psychrotrophic, coliform, and gram-positive catalase-negative cocci (GPCNC) bacterial groups. Thermoduric, psychrotrophic, and coliform species were significantly related to BTTBC, whereas GPCNC were correlated with both BTTBC and BTSCC variables. Highest counts were for psychrotroph and coliform groups, and a moderate to high correlation (r=0.51) was found between both variables, indicating that poor cleaning practices in the flocks tend to select for less-resistant organisms, such as gram-negative rods. In addition, BTTBC correlated with BTSCC (r=0.42). Some variation factors for specific bacterial counts, such as breed, season, milking type, dry therapy, and milk yield, were also analyzed. Flock information was collected from flock books, annual audits, and the CPO traceability system. Psychrotrophs and coliforms had elevated counts in winter, whereas GPCNC were higher in summer and in hand-milked flocks. Dry therapy contributed to the reduction in psychrotrophic bacteria; therefore, some strains of mammary pathogens could also be psychrotrophic bacteria. Results of this study would be helpful for troubleshooting milk quality problems and developing premium payment systems in dairy sheep. PMID:23200475

  6. Relationship of somatic cell count, physical, chemical and enzymatic properties to the bacterial standard plate count in dairy goat milk

    Microsoft Academic Search

    Chingwen Ying; Han-Tsung Wang; Jih-Tay Hsu

    2002-01-01

    Weekly milk samples, collected from individual half mammary gland of three late lactation (experiment 1) and four early lactation (experiment 2) Alpine dairy goats in second lactation, were subjected to various physical and chemical measurements to examine the relationship of the logarithm of bacterial standard plate count (SPC) to electrical conductivity (EC), neutrophils\\/lymphocytes ratio, lactate dehydrogenase (LDH), tyrosine concentration, alkaline

  7. The relationship between absolute lymphocyte count with PFS in patients with Hodgkin's lymphoma undergoing autologous hematopoietic cell transplant

    Microsoft Academic Search

    T Seshadri; M Pintilie; A Keating; M Crump; J Kuruvilla

    2008-01-01

    Previous reports in Hodgkin's lymphoma (HL) patients undergoing autologous hematopoietic cell transplantation (AHCT) have demonstrated a significant association between the absolute lymphocyte count at day 15 (ALC-15) with survival. To evaluate this finding further, we analyzed 146 patients with relapsed\\/refractory HL who underwent AHCT to evaluate the relationship between lymphocyte counts at apheresis and at two time points (days 15

  8. A microfluidic device for practical label-free CD4+ T cell counting of HIV-infected subjects

    E-print Network

    Demirci, Utkan

    counting of a particular white blood cell subset, the CD4+ T lymphocyte. To address the limitations shear flow to specifically isolate CD4+ T lymphocytes with high efficiency directly from 10 microlitres cytometry among HIV-positive subjects over a wide range of absolute CD4 counts (R2 = 0.93). This CD4

  9. Comparison of Two Methods for the Determination of the Effects of Ionizing Radiation on Blood Cell Counts in Mice

    PubMed Central

    Romero-Weaver, Ana L.; Kennedy, Ann R.

    2012-01-01

    A reliable technique is needed to determine the effect of ionizing radiation on white blood cell (WBC) counts. Facilities that utilize automated methods can provide this service. However, utilizing external facilities can introduce additional variables, such as differences between time of sample collection and time of sample processing, which may affect the results. The purpose of the present study was to determine whether an automated method at an external facility can accurately determine radiation-induced changes in total WBC, lymphocyte and granulocyte counts when samples are analyzed at periods of time up to 24 hours after collection and stored either at room temperature or at 4°C. To accomplish this, we compared automated blood cell counts determined at an external facility with our manual blood cell counts processed immediately after sample collection or 24 h after sample collection and stored either at room temperature or 4°C from mice exposed to 2 Gy proton or 2 Gy gamma radiation. Our results show a close correlation and good agreement between the two methods, indicating that neither a delay of 24 hours in sample processing nor storage temperature affected white blood cell counts. Analysis of the effects of radiation on blood cell counts by either manual or automated cell counts revealed a statistically significant decrease in lymphocyte and granulocyte counts at different days post-irradiation, with no statistically significant difference between the methods employed; therefore both manual and automated blood cell counts are reliable methods to determine the effects of ionizing radiation in blood cells. PMID:23450807

  10. Measurement of radionuclides using ion chromatography and flow-cell scintillation counting with pulse shape discrimination

    SciTech Connect

    DeVol, T.A.; Fjeld, R.A.

    1995-12-31

    A project has been initiated at Clemson Univ. to develop a HPLC/flow- cell system for analysis of non-gamma emitting radionuclides in environmental samples; an important component is development of a low background flow-cell detector that counts alpha and beta particles separately through pulse shape discrimination. Objective of the work presented here is to provide preliminary results of an evaluation of the following scintillators: CaF{sub 2}:Eu, scintillating glass, and BaF{sub 2}. Slightly acidic aqueous solutions of the alpha emitter {sup 233}U and the beta emitter {sup 45}Ca were used. Detection efficiencies and minimum detectable activities were determined.

  11. Correlation of striatal dopamine transporter imaging with post mortem substantia nigra cell counts.

    PubMed

    Kraemmer, Julia; Kovacs, Gabor G; Perju-Dumbrava, Laura; Pirker, Susanne; Traub-Weidinger, Tatiana; Pirker, Walter

    2014-12-01

    Dopamine transporter imaging is widely used for the differential diagnosis of parkinsonism. Only limited data are available on the relationship between striatal dopamine transporter binding and dopaminergic cell loss in the substantia nigra (SN). We analyzed postmortem SN cell counts in patients who had previously undergone dopamine transporter single-photon emission computed tomography (SPECT). Pathological diagnoses included Parkinson's disease (n?=?1), dementia with Lewy bodies (n?=?2), multiple system atrophy (n?=?1), corticobasal degeneration (n?=?2), atypical parkinsonism with multiple pathological conditions (n?=?1), Alzheimer's disease (n?=?1), and Creutzfeldt-Jakob disease (n?=?1). [(12) (3) I]?-CIT SPECT had been performed in all subjects using a standardized protocol on the same triple-head gamma camera. The density of neuromelanin-containing and tyrosine hydroxylase-positive substantia nigra neurons/mm(2) was evaluated in paraffin-embedded tissue sections by morphometric methods. Mean disease duration at the time of dopamine transporter imaging was 2.3 years, and the mean interval from imaging to death was 29.3 months (range, 4-68 months). Visual analysis of dopamine transporter images showed reduced striatal uptake in all seven patients with neurodegenerative parkinsonism, but not in Alzheimer's and Creutzfeldt-Jakob disease cases. Averaged [(right+left)/2] striatal uptake was highly correlated with averaged SN cell counts (rs ?=?0.98, P?cells). Similar strong correlations were found in separate analyses for the right and left sides. Striatal dopamine transporter binding highly correlated with postmortem SN cell counts, confirming the validity of dopamine transporter imaging as an excellent in vivo marker of nigrostriatal dopaminergic degeneration. PMID:25048738

  12. Fluorescence based cell counting in collagen monolayer cultures of primary hepatocytes.

    PubMed

    Priesnitz, C; Sperber, S; Garg, R; Orsini, M; Noor, F

    2014-11-26

    Accurate determination of cell number is essential for the quantitative description of biological processes. The changes should be related to a measurable reference e.g. in the case of cell culture, the viable cell number is a very valuable reference parameter. Indirect methods of cell number/viability measurements may have up to 10 % standard deviation. This can lead to undesirable large deviations in the analysis of "-omics" data as well as time course studies. Such data should be preferably normalized to the exact viable cell number at a given time to allow meaningful interpretation and understanding of the biological processes. Manual counting of cell number is very laborious and not possible in certain experimental setups. We therefore, developed a simple and reliable fluorescence based method with an accuracy of 95-98 % for the determination of the viable cell number in situ. We optimized the seeding cell densities for primary rat hepatocytes for optimal cell adhesion. This will help in efficient use of primary cells which are usually limited in availability. The method will be very useful in the application of "-omics" techniques, especially metabolome analysis where the specific rates of uptake/production of metabolites can be reliably calculated. PMID:25424145

  13. Analysis of White Blood Cell Counts in Mice after Gamma- or Proton-Radiation Exposure

    PubMed Central

    Maks, Casey J.; Wan, X. Steven; Ware, Jeffrey H.; Romero-Weaver, Ana L.; Sanzari, Jenine K.; Wilson, Jolaine M.; Rightnar, Steve; Wroe, Andrew J.; Koss, Peter; Gridley, Daila S.; Slater, James M.; Kennedy, Ann R.

    2013-01-01

    In the coming decades human space exploration is expected to move beyond low-Earth orbit. This transition involves increasing mission time and therefore an increased risk of radiation exposure from solar particle event (SPE) radiation. Acute radiation effects after exposure to SPE radiation are of prime importance due to potential mission-threatening consequences. The major objective of this study was to characterize the dose–response relationship for proton and ? radiation delivered at doses up to 2 Gy at high (0.5 Gy/min) and low (0.5 Gy/h) dose rates using white blood cell (WBC) counts as a biological end point. The results demonstrate a dose-dependent decrease in WBC counts in mice exposed to high- and low-dose-rate proton and ? radiation, suggesting that astronauts exposed to SPE-like radiation may experience a significant decrease in circulating leukocytes. PMID:21476859

  14. Short Communication: Bulk Tank Total Bacterial Count in Dairy Sheep: Factors of Variation and Relationship with Somatic Cell Count

    Microsoft Academic Search

    C. Gonzalo; J. A. Carriedo; E. Beneitez; M. T. Juárez; L. F. De La Fuente; F. San Primitivo

    2006-01-01

    A total of 9,353 records for bulk tank total bacterial count (TBC) were obtained over 1 yr from 315 dairy ewe flocks belonging to the Sheep Improvement Consortium (CPO) in Castilla-Leon (Spain). Analysis of variance showed significant effects of flock, breed, month within flock, dry therapy, milking type and installation, and logSCC on logTBC. Flock and month within flock were

  15. Intramammary infections and risk factors for clinical mastitis in herds with low somatic cell counts in bulk milk

    Microsoft Academic Search

    YH Schukken; D Van de Geer; FJ Grommers; JA Smit; A Brand

    1989-01-01

    Ten herds with low somatic cell counts in bulk milk had an incidence of clinical mastitis of only 2.2 per 100 cows whereas 10 other herds with similarly low cell counts had an incidence of 53.6 per 100 cows. The major pathogens in the herds with a high incidence were Escherichia coli, Streptococcus uberis, Staphylococcus aureus and the coagulase-negative staphylococci.

  16. Effect of season on milk temperature, milk growth hormone, prolactin, and somatic cell counts of lactating cattle

    NASA Astrophysics Data System (ADS)

    Igono, M. O.; Johnson, H. D.; Steevens, B. J.; Hainen, W. A.; Shanklin, M. D.

    1988-09-01

    Monthly fluctuations in milk temperature, somatic cell counts, milk growth hormone and prolactin of lactating cows were measured in milk samples over a 1 year period. The seasonal patterns in milk temperature, somatic cell count and milk prolactin concentration showed a positive trend with increasing environmental temperatures. Milk growth hormone concentration increased with lactation level and declined significantly during summer heat. Milk temperature and the measured hormonal levels may serve as indicators of the impact of the climatic environment on lactating cattle.

  17. Somatic cell counts determined by the Coulter or Fossomatic Counter and their relationship to administration of oxytocin

    Microsoft Academic Search

    A. R Burriel

    1999-01-01

    The somatic cell count of milk from experimentally inoculated and control mammary glands of primiparous ewes from meat-type breeds were determined by Coulter and Fossomatic Counters. Oxytocin was administered prior to milk collection to avoid stripping of the mammary epithelium while assuring sufficient quantity of milk, but was found to increase (***p<0.0001) the somatic cell counts when they were determined

  18. Effects of season, milking routine and cow cleanliness on bacterial and somatic cell counts of bulk tank milk.

    PubMed

    Zucali, Maddalena; Bava, Luciana; Tamburini, Alberto; Brasca, Milena; Vanoni, Laura; Sandrucci, Anna

    2011-11-01

    The aim of the study was to investigate the effects of season, cow cleanliness and milking routine on bacterial and somatic cell counts of bulk tank milk. A total of 22 dairy farms in Lombardy (Italy) were visited three times in a year in different seasons. During each visit, samples of bulk tank milk were taken for bacterial and somatic cell counts; swabs from the teat surface of a group of cows were collected after teat cleaning and before milking. Cow cleanliness was assessed by scoring udder, flanks and legs of all milking cows using a 4-point scale system. Season affected cow cleanliness with a significantly higher percentage of non-clean (NC) cows during Cold compared with Mild season. Standard plate count (SPC), laboratory pasteurization count (LPC), coliform count (CC) and somatic cell count, expressed as linear score (LS), in milk significantly increased in Hot compared with Cold season. Coagulase-positive staphylococci on teat swabs showed higher counts in Cold season in comparison with the other ones. The effect of cow cleanliness was significant for SPC, psychrotrophic bacterial count (PBC), CC and Escherichia coli in bulk tank milk. Somatic cell count showed a relationship with udder hygiene score. Milking operation routine strongly affected bacterial counts and LS of bulk tank milk: farms that accomplished a comprehensive milking scheme including two or more operations among forestripping, pre-dipping and post-dipping had lower teat contamination and lower milk SPC, PBC, LPC, CC and LS than farms that did not carry out any operation. PMID:21843397

  19. CD4 T cell survival after intermittent interleukin-2 therapy is predictive of an increase in the CD4 T cell count of HIV-infected patients.

    PubMed

    Read, Sarah W; Lempicki, Richard A; Di Mascio, Michele; Srinivasula, Sharat; Burke, Rosanne; Sachau, William; Bosche, Marjorie; Adelsberger, Joseph W; Sereti, Irini; Davey, Richard T; Tavel, Jorge A; Huang, Chiung-Yu; Issaq, Haleem J; Fox, Stephen D; Lane, H Clifford; Kovacs, Joseph A

    2008-09-15

    Administration of interleukin (IL)-2 to human immunodeficiency virus (HIV)-infected patients leads to significant increases in CD4 T cell counts. We previously have shown that IL-2 induces increased proliferation and survival of CD4 T cells. Deuterium labeling studies were undertaken to study the relationship between IL-2-induced increases in the CD4 T cell count and the effects of IL-2 on cell proliferation and survival. A strong inverse correlation was noted between the rate of decay of the label in CD4 cells and increases in CD4 cell counts (R =or- 0.67; P<.001). This correlation was not seen with the level of proliferating cells. Although the CD4 cell count at baseline and the number of CD4 cells expressing CD25 were also predictive of increases in the CD4 cell count, the rate of decay remained the most statistically significant predictor in multivariate regression models. Thus, an increase in the survival of CD4 T cells appears to be the critical mechanism leading to sustained increases in the CD4 cell counts of HIV-infected patients receiving intermittent IL-2 therapy. PMID:18684102

  20. Recruitment of host’s progenitor cells to sites of human amniotic fluid stem cells implantation

    Microsoft Academic Search

    Teodelinda Mirebella; Alessandro Poggi; Monica Scaranari; Massimo Mogni; Mario Lituania; Chiara Baldo; Ranieri Cancedda; Chiara Gentili

    2011-01-01

    The amniotic fluid is a new source of multipotent stem cells with a therapeutic potential for human diseases. Cultured at low cell density, human amniotic fluid stem cells (hAFSCs) were still able to generate colony-forming unit-fibroblast (CFU-F) after 60 doublings, thus confirming their staminal nature. Moreover, after extensive in vitro cell expansion hAFSCs maintained a stable karyotype. The expression of

  1. Design, fabrication, and applications of in situ fluid cell TEM.

    PubMed

    Li, Dongsheng; Nielsen, Michael H; De Yoreo, James J

    2013-01-01

    In situ fluid cell TEM is a powerful new tool for understanding dynamic processes during liquid phase chemical reactions, including mineral formation. This technique, which operates in the high vacuum of a TEM chamber, provides information on crystal structure, phase, morphology, size, aggregation/segregation, and crystal growth mechanisms in real time. In situ TEM records both crystal structure and morphology at spatial resolutions down to the atomic level with high temporal resolution of up to 10(-6)s per image, giving it distinct advantages over other in situ techniques such as optical microscopy, AFM, or X-ray scattering or diffraction. This chapter addresses the design, fabrication, and assembly of TEM fluid cells and applications of fluid cell TEM to understanding mechanisms of mineralization. PMID:24188766

  2. Fluid dynamics and noise in bacterial cell–cell and cell–surface scattering

    PubMed Central

    Drescher, Knut; Dunkel, Jörn; Cisneros, Luis H.; Ganguly, Sujoy; Goldstein, Raymond E.

    2011-01-01

    Bacterial processes ranging from gene expression to motility and biofilm formation are constantly challenged by internal and external noise. While the importance of stochastic fluctuations has been appreciated for chemotaxis, it is currently believed that deterministic long-range fluid dynamical effects govern cell–cell and cell–surface scattering—the elementary events that lead to swarming and collective swimming in active suspensions and to the formation of biofilms. Here, we report direct measurements of the bacterial flow field generated by individual swimming Escherichia coli both far from and near to a solid surface. These experiments allowed us to examine the relative importance of fluid dynamics and rotational diffusion for bacteria. For cell–cell interactions it is shown that thermal and intrinsic stochasticity drown the effects of long-range fluid dynamics, implying that physical interactions between bacteria are determined by steric collisions and near-field lubrication forces. This dominance of short-range forces closely links collective motion in bacterial suspensions to self-organization in driven granular systems, assemblages of biofilaments, and animal flocks. For the scattering of bacteria with surfaces, long-range fluid dynamical interactions are also shown to be negligible before collisions; however, once the bacterium swims along the surface within a few microns after an aligning collision, hydrodynamic effects can contribute to the experimentally observed, long residence times. Because these results are based on purely mechanical properties, they apply to a wide range of microorganisms. PMID:21690349

  3. Straw blood cell count, growth, inhibition and comparison to apoptotic bodies

    PubMed Central

    Wu, Yonnie; Henry, David C; Heim, Kyle; Tomkins, Jeffrey P; Kuan, Cheng-Yi

    2008-01-01

    Background Mammalian cells transform into individual tubular straw cells naturally in tissues and in response to desiccation related stress in vitro. The transformation event is characterized by a dramatic cellular deformation process which includes: condensation of certain cellular materials into a much smaller tubular structure, synthesis of a tubular wall and growth of filamentous extensions. This study continues the characterization of straw cells in blood, as well as the mechanisms of tubular transformation in response to stress; with specific emphasis placed on investigating whether tubular transformation shares the same signaling pathway as apoptosis. Results There are approximately 100 billion, unconventional, tubular straw cells in human blood at any given time. The straw blood cell count (SBC) is 45 million/ml, which accounts for 6.9% of the bloods dry weight. Straw cells originating from the lungs, liver and lymphocytes have varying nodules, hairiness and dimensions. Lipid profiling reveals severe disruption of the plasma membrane in CACO cells during transformation. The growth rates for the elongation of filaments and enlargement of rabbit straw cells is 0.6~1.1 (?m/hr) and 3.8 (?m3/hr), respectively. Studies using apoptosis inhibitors and a tubular transformation inhibitor in CACO2 cells and in mice suggested apoptosis produced apoptotic bodies are mediated differently than tubular transformation produced straw cells. A single dose of 0.01 mg/kg/day of p38 MAPK inhibitor in wild type mice results in a 30% reduction in the SBC. In 9 domestic animals SBC appears to correlate inversely with an animal's average lifespan (R2 = 0.7). Conclusion Straw cells are observed residing in the mammalian blood with large quantities. Production of SBC appears to be constant for a given animal and may involve a stress-inducible protein kinase (P38 MAPK). Tubular transformation is a programmed cell survival process that diverges from apoptosis. SBCs may be an important indicator of intrinsic aging-related stress. PMID:18492269

  4. Computer-assisted counting of retinal cells by automatic segmentation after TV denoising

    PubMed Central

    2013-01-01

    Background Quantitative evaluation of mosaics of photoreceptors and neurons is essential in studies on development, aging and degeneration of the retina. Manual counting of samples is a time consuming procedure while attempts to automatization are subject to various restrictions from biological and preparation variability leading to both over- and underestimation of cell numbers. Here we present an adaptive algorithm to overcome many of these problems. Digital micrographs were obtained from cone photoreceptor mosaics visualized by anti-opsin immuno-cytochemistry in retinal wholemounts from a variety of mammalian species including primates. Segmentation of photoreceptors (from background, debris, blood vessels, other cell types) was performed by a procedure based on Rudin-Osher-Fatemi total variation (TV) denoising. Once 3 parameters are manually adjusted based on a sample, similarly structured images can be batch processed. The module is implemented in MATLAB and fully documented online. Results The object recognition procedure was tested on samples with a typical range of signal and background variations. We obtained results with error ratios of less than 10% in 16 of 18 samples and a mean error of less than 6% compared to manual counts. Conclusions The presented method provides a traceable module for automated acquisition of retinal cell density data. Remaining errors, including addition of background items, splitting or merging of objects might be further reduced by introduction of additional parameters. The module may be integrated into extended environments with features such as 3D-acquisition and recognition. PMID:24138794

  5. Clinical judgement of airway inflammation versus sputum cell counts in patients with asthma

    Microsoft Academic Search

    K. Parameswaran; E. Pizzichini; M. M. Pizzichini; P. Hussack; A. Efthimiadis; F. E. Hargreave

    2000-01-01

    ABSTRACT: The inflammatory,component,of asthma,is usually assessed indirectly by symptoms and spirometry, these may be inaccurate. It can now be assessed directly and reliably by the examination,of sputum,cell counts. There is no information,on how clinical assessment,of the presence,and,type of airway,inflammation,compares,with actual measurements. In this single-centre observational study, sputum was collected from 76 consecutive adults with asthma,attending a tertiary chest clinic after

  6. Fluid transport by cultured corneal epithelial cell layers

    PubMed Central

    Yang, H; Reinach, P; Koniarek, J; Wang, Z; Iserovich, P; Fischbarg, J

    2000-01-01

    BACKGROUND/AIMS—Fluid transport across the in vitro corneal epithelium is short lived, hence difficult to detect and characterise. Since stable rates of fluid transport across several cultured epithelial cell layers have been demonstrated, the behaviour of confluent SV40 transformed rabbit corneal epithelial cells (tRCEC) grown on permeable supports was examined.?METHODS—Fluid transport was determined with a nanoinjector volume clamp; the specific electrical resistance of the layers was 184 (SEM 9) ? cm2. tRCEC layers transported fluid (from basal to apical) against a pressure head of 3 cm H2O for 2-3 hours.?RESULTS—In the first hour, the rate of fluid transport was 5.2 (0.5) µl/h/cm?2 (n=23), which is comparable with that found in other epithelia. Fluid transport was completely inhibited in 15-30 minutes by either 100 µM ouabain (n=6), 50 µM bumetanide (n=6), or 1 µM endothelin-1 (ET-1; n=6). Preincubation with 10 µM BQ123 (an ETA receptor antagonist) obviated inhibition by ET-1 (n=6). ET-1 also caused a 22% decrease in specific resistance.?CONCLUSIONS—Fluid transport appears to depend on transepithelial Cl transport since (1) their directions are the same (stroma?tear), and (2) both bumetanide and ouabain inhibit it with similar time course. tRCEC appear useful to investigate aspects of the physiology and pharmacology of fluid transport across this layer, including receptor mediated control of this process.?? PMID:10655198

  7. Influence of NK cell count on the survival of patients with diffuse large B-cell lymphoma treated with R-CHOP

    PubMed Central

    Kim, Joong-Keun; Shin, Ho-Jin; Song, Moo-Kon; Yi, Ji-Won; Shin, Dong-Hun; Lee, Dae-Sung; Baek, Sung-Min

    2014-01-01

    Background Although adding rituximab to the chemotherapy regimen of cyclophosphamide, vincristine, doxorubicin, and prednisone (R-CHOP) has improved clinical outcomes of patients with diffuse large B-cell lymphoma (DLBCL), several recent studies have shown that the effect of rituximab is dominantly in the non-germinal center (non-GC) subtype compared to the germinal center (GC) subtype. Natural killer (NK) cell count, a surrogate marker of immune status, is associated with clinical outcomes in DLBCL patients in the rituximab era. We investigated whether the impact of NK cells on clinical outcomes differed according to the immunophenotype of DLBCL. Methods This study analyzed 72 DLBCL patients treated with R-CHOP between January 2010 and January 2014. Results Low NK cell counts (<100/µL) were associated with poor progression-free survival (PFS) and overall survival (OS) compared to high NK cell counts. In multivariate analysis, low NK cell count was an independent prognostic factor for PFS and OS. However, survival did not significantly differ between the GC and non-GC subtypes. We examined the clinical influence of NK cells according to the immunophenotype and found that low NK cell counts were significantly associated with poor PFS and OS in non-GC cases, but not in GC cases. Conclusion Low NK cell counts at diagnosis are associated with poor clinical outcomes in DLBCL patients treated with R-CHOP therapy. However, the impact is significant only in non-GC subtype DLBCL, not in the GC subtype. PMID:25325035

  8. Effect of coagulase-negative staphylococci on somatic cell count in Dutch dairy herds.

    PubMed

    Sampimon, Otlis; van den Borne, Bart Hp; Santman-Berends, Inge; Barkema, Herman W; Lam, Theo

    2010-08-01

    The effect was quantified of coagulase-negative staphylococci (CNS) intramammary infections on quarter- and cow-level somatic cell count (SCC) and on bulk milk somatic cell count (BMSCC) in different BMSCC cohorts in Dutch dairy herds. Two datasets were used for this purpose. In the first dataset, on 49 randomly selected dairy farms a total of 4220 quarter milk samples of 1072 cows were collected of all cows and heifers with a test-day SCC 250 000 and 150 000 cells/ml, respectively, and of 25% of cows and heifers below these thresholds. In the second dataset, on 39 selected dairy farms a total of 8329 quarter milk samples of 2115 cows were collected of all cows with a test-day SCC 250 000 cells/ml following two consecutive SCC <250 000 cells/ml, and of heifers using the same SCC criteria but with a threshold of 150 000 cells/ml. These cows and heifers were defined as new high SCC. In both datasets, CNS was the most frequently isolated pathogen, 11% in the first dataset and 12% in the second dataset. In both datasets, quarters with CNS IMI had a lower SCC than quarters infected with major pathogens, and a higher SCC than culture-negative quarters. The same was found for SCC at cow level. Coagulase-negative staphylococci were more often found in quarters with SCC 200 000 cells/ml in dairy farms with a BMSCC <150 000 cells/ml compared with dairy farms with a higher BMSCC. Prevalence of CNS in cows and heifers with a high SCC was higher in dairy farms with a BMSCC <150 000 cells/ml compared with dairy farms with a medium or high BMSCC: 30, 19 and 18%, respectively. This indicates that CNS IMI as a cause of subclinical mastitis is relatively more important in dairy farms with a low BMSCC and may become a point of attention in udder health management on that type of farm. PMID:20450528

  9. Fluid transport by cultured corneal epithelial cell layers

    Microsoft Academic Search

    H Yang; P S Reinach; J P Koniarek; Z Wang; P Iserovich; J Fischbarg

    2000-01-01

    BACKGROUND\\/AIMSFluid transport across the in vitro corneal epithelium is short lived, hence difficult to detect and characterise. Since stable rates of fluid transport across several cultured epithelial cell layers have been demonstrated, the behaviour of confluent SV40 transformed rabbit corneal epithelial cells (tRCEC) grown on permeable supports was examined.METHODSFluid transport was determined with a nanoinjector volume clamp; the specific electrical

  10. Benchmarks for evaluation and comparison of udder health status using monthly individual somatic cell count

    PubMed Central

    Fauteux, Véronique; Roy, Jean-Philippe; Scholl, Daniel T.; Bouchard, Émile

    2014-01-01

    The objectives of this study were to propose benchmarks for the interpretation of herd udder health using monthly individual somatic cell counts (SCC) from dairy herds in Quebec, Canada and to evaluate the association of risk factors with intramammary infection (IMI) dynamics relative to these benchmarks. The mean and percentiles of indices related to udder infection status [e.g., proportion of healthy or chronically infected cows, cows cured and new IMI (NIMI) rate] during lactation and over the dry period were calculated using a threshold of ? 200 000 cells/mL at test day. Mean NIMI proportion and proportion of cows cured during lactation were 0.11 and 0.27. Benchmarks of 0.70 and 0.03 for healthy and chronically infected cows over the dry period were proposed. Season and herd mean SCC were risk factors influencing IMI dynamics during lactation and over the dry period. PMID:25082989

  11. Modeling Centrifugal Cell Washers Using Computational Fluid Dynamics

    Microsoft Academic Search

    Beth E. Kellet; Binbing Han; David S. Dandy; S. Ranil Wickramasinghe

    2004-01-01

    Reinfusion of shed blood during surgery could avoid the need for blood transfusions. Prior to reinfusion of the red blood cells, the shed blood must be washed in order to remove leukocytes, platelets, and other contami- nants. Further, the hematocrit of the washed blood must be increased. The feasibility of using computational fluid dynamics (CFD) to guide the design of

  12. Neutron confinement cell for investigating complex fluids Tonya L. Kuhla)

    E-print Network

    Kuhl, Tonya L.

    Neutron confinement cell for investigating complex fluids Tonya L. Kuhla) Department of Chemical Manuel Lujan, Jr. Neutron Scattering Center, LANSCE-12, MS H805, Los Alamos National Laboratory, Los Lujan, Jr. Neutron Scattering Center, LANSCE-12, MS H805, Los Alamos National Laboratory, Los Alamos

  13. Absolute Lymphocyte Count on Day 30 Is a Surrogate for Robust Hematopoietic Recovery and Strongly Predicts Outcome after T Cell-Depleted Allogeneic Stem Cell Transplantation

    Microsoft Academic Search

    Bipin N. Savani; Stephan Mielke; Katayoun Rezvani; Aldemar Montero; Agnes S. Yong; Laura Wish; Jeannine Superata; Roger Kurlander; Anurag Singh; Richard Childs; A. John Barrett

    2007-01-01

    Several studies have shown that a higher lymphocyte count 3-4 weeks after allogeneic stem cell transplantation (SCT) is associated with better transplant outcome. However, the factors determining early lymphocyte recovery are not defined. To further explore the relationship between lymphocyte recovery and outcome we analyzed lymphocyte counts and other engraftment parameters in 157 patients with leukemia (48 acute myelogenous leukemia,

  14. Validation of analytical methods in GMP: the disposable Fast Read 102® device, an alternative practical approach for cell counting

    PubMed Central

    2012-01-01

    Background The quality and safety of advanced therapy products must be maintained throughout their production and quality control cycle to ensure their final use in patients. We validated the cell count method according to the International Conference on Harmonization of Technical Requirements for Registration of Pharmaceuticals for Human Use and European Pharmacopoeia, considering the tests’ accuracy, precision, repeatability, linearity and range. Methods As the cell count is a potency test, we checked accuracy, precision, and linearity, according to ICH Q2. Briefly our experimental approach was first to evaluate the accuracy of Fast Read 102® compared to the Bürker chamber. Once the accuracy of the alternative method was demonstrated, we checked the precision and linearity test only using Fast Read 102®. The data were statistically analyzed by average, standard deviation and coefficient of variation percentages inter and intra operator. Results All the tests performed met the established acceptance criteria of a coefficient of variation of less than ten percent. For the cell count, the precision reached by each operator had a coefficient of variation of less than ten percent (total cells) and under five percent (viable cells). The best range of dilution, to obtain a slope line value very similar to 1, was between 1:8 and 1:128. Conclusions Our data demonstrated that the Fast Read 102® count method is accurate, precise and ensures the linearity of the results obtained in a range of cell dilution. Under our standard method procedures, this assay may thus be considered a good quality control method for the cell count as a batch release quality control test. Moreover, the Fast Read 102® chamber is a plastic, disposable device that allows a number of samples to be counted in the same chamber. Last but not least, it overcomes the problem of chamber washing after use and so allows a cell count in a clean environment such as that in a Cell Factory. In a good manufacturing practice setting the disposable cell counting devices will allow a single use of the count chamber they can then be thrown away, thus avoiding the waste disposal of vital dye (e.g. Trypan Blue) or lysing solution (e.g. Tuerk solution). PMID:22650233

  15. PCR-Based Multiple Species Cell Counting for In Vitro Mixed Culture

    PubMed Central

    Huang, Ruijie; Zhang, Junjie; Yang, X. Frank; Gregory, Richard L.

    2015-01-01

    Changes of bacterial profiles in microbial communities are strongly associated with human health. There is an increasing need for multiple species research in vitro. To avoid high cost or measurement of a limited number of species, PCR-based multiple species cell counting (PCR-MSCC) has been conceived. Species-specific sequence is defined as a unique sequence of one species in a multiple species mixed culture. This sequence is identified by comparing a random 1000 bp genomic sequence of one species with the whole genome sequences of the other species in the same artificial mixed culture. If absent in the other genomes, it is the species-specific sequence. Species-specific primers were designed based on the species-specific sequences. In the present study, ten different oral bacterial species were mixed and grown in Brain Heart Infusion Yeast Extract with 1% sucrose for 24 hours. Biofilm was harvested and processed for DNA extraction and q-PCR amplification with the species-specific primers. By comparing the q-PCR data of each species in the unknown culture with reference cultures, in which the cell number of each species was determined by colony forming units on agar plate, the cell number of that strain in the unknown mixed culture was calculated. This technique is reliable to count microorganism numbers that are less than 100,000 fold different from other species within the same culture. Theoretically, it can be used in detecting a species in a mixed culture of over 200 species. Currently PCR-MSCC is one of the most economic methods for quantifying single species cell numbers, especially for the low abundant species, in a multiple artificial mixed culture in vitro. PMID:25970462

  16. Smoking status and differential white cell count in men and women in the EPIC-Norfolk population

    Microsoft Academic Search

    Megan R. Smith; Ann-Louise Kinmonth; Robert N. Luben; Sheila Bingham; Nicholas E. Day; Nicholas J. Wareham; Ailsa Welch; Kay-Tee Khaw

    2003-01-01

    The total white blood cell (WBC) count is reported to be an independent predictor of mortality in several prospective studies. We investigated the association between total and differential WBC counts and cigarette smoking habit in a cross-sectional population-based study of 6902 men and 8405 women 39–79 years of age participating between July 1994 and 1997 in the European Prospective Investigation

  17. Amniotic Fluid-Derived Stem Cells for Cardiovascular Tissue Engineering Applications

    E-print Network

    Petsche Connell, Jennifer

    Recent research has demonstrated that a population of stem cells can be isolated from amniotic fluid removed by amniocentesis that are broadly multipotent and nontumorogenic. These amniotic fluid-derived stem cells (AFSC) ...

  18. Fluid phase biopsy for detection and characterization of circulating endothelial cells in myocardial infarction

    PubMed Central

    Bethel, Kelly; Luttgen, Madelyn S; Damani, Samir; Kolatkar, Anand; Lamy, Rachelle; Sabouri-Ghomi, Mohsen; Topol, Sarah; Topol, Eric J; Kuhn, Peter

    2014-01-01

    Elevated levels of circulating endothelial cells (CECs) occur in response to various pathological conditions including myocardial infarction (MI). Here, we adapted a fluid phase biopsy technology platform that successfully detects circulating tumor cells in blood of cancer patients (HD-CTC assay), to create a High-Definition Circulating Endothelial Cell (HD-CEC) assay for the detection and characterization of CECs. Peripheral blood samples were collected from 79 MI patients, 25 healthy controls and 6 patients undergoing vascular surgery (VS). CECs were defined by positive staining for DAPI, CD146 and von Willebrand Factor and negative staining for CD45. In addition, CECs exhibited distinct morphological features that enable differentiation from surrounding white blood cells. CECs were found both as individual cells and as aggregates. CEC numbers were higher in MI patients compared with healthy controls. VS patients had lower CEC counts when compared with MI patients but were not different from healthy controls. Both HD-CEC and CellSearch® assays could discriminate MI patients from healthy controls with comparable accuracy but the HD-CEC assay exhibited higher specificity while maintaining high sensitivity. Our HD-CEC assay may be used as a robust diagnostic biomarker in MI patients. PMID:24406475

  19. Fluid phase biopsy for detection and characterization of circulating endothelial cells in myocardial infarction.

    PubMed

    Bethel, Kelly; Luttgen, Madelyn S; Damani, Samir; Kolatkar, Anand; Lamy, Rachelle; Sabouri-Ghomi, Mohsen; Topol, Sarah; Topol, Eric J; Kuhn, Peter

    2014-02-01

    Elevated levels of circulating endothelial cells (CECs) occur in response to various pathological conditions including myocardial infarction (MI). Here, we adapted a fluid phase biopsy technology platform that successfully detects circulating tumor cells in the blood of cancer patients (HD-CTC assay), to create a high-definition circulating endothelial cell (HD-CEC) assay for the detection and characterization of CECs. Peripheral blood samples were collected from 79 MI patients, 25 healthy controls and six patients undergoing vascular surgery (VS). CECs were defined by positive staining for DAPI, CD146 and von Willebrand Factor and negative staining for CD45. In addition, CECs exhibited distinct morphological features that enable differentiation from surrounding white blood cells. CECs were found both as individual cells and as aggregates. CEC numbers were higher in MI patients compared with healthy controls. VS patients had lower CEC counts when compared with MI patients but were not different from healthy controls. Both HD-CEC and CellSearch® assays could discriminate MI patients from healthy controls with comparable accuracy but the HD-CEC assay exhibited higher specificity while maintaining high sensitivity. Our HD-CEC assay may be used as a robust diagnostic biomarker in MI patients. PMID:24406475

  20. Fluid phase biopsy for detection and characterization of circulating endothelial cells in myocardial infarction

    NASA Astrophysics Data System (ADS)

    Bethel, Kelly; Luttgen, Madelyn S.; Damani, Samir; Kolatkar, Anand; Lamy, Rachelle; Sabouri-Ghomi, Mohsen; Topol, Sarah; Topol, Eric J.; Kuhn, Peter

    2014-02-01

    Elevated levels of circulating endothelial cells (CECs) occur in response to various pathological conditions including myocardial infarction (MI). Here, we adapted a fluid phase biopsy technology platform that successfully detects circulating tumor cells in the blood of cancer patients (HD-CTC assay), to create a high-definition circulating endothelial cell (HD-CEC) assay for the detection and characterization of CECs. Peripheral blood samples were collected from 79 MI patients, 25 healthy controls and six patients undergoing vascular surgery (VS). CECs were defined by positive staining for DAPI, CD146 and von Willebrand Factor and negative staining for CD45. In addition, CECs exhibited distinct morphological features that enable differentiation from surrounding white blood cells. CECs were found both as individual cells and as aggregates. CEC numbers were higher in MI patients compared with healthy controls. VS patients had lower CEC counts when compared with MI patients but were not different from healthy controls. Both HD-CEC and CellSearch® assays could discriminate MI patients from healthy controls with comparable accuracy but the HD-CEC assay exhibited higher specificity while maintaining high sensitivity. Our HD-CEC assay may be used as a robust diagnostic biomarker in MI patients.

  1. Farm management factors associated with bulk tank somatic cell count in Irish dairy herds

    PubMed Central

    2009-01-01

    The relationship between bulk tank somatic cell count (SCC) and farm management and infrastructure was examined using data from 398 randomly selected, yet representative, Irish dairy farms where the basal diet is grazed grass. Median bulk tank SCC for the farms was 282,887 cells/ml ranging from 82,209 to 773,028 cells/ml. Two questionnaires were administered through face-to-face contact with each farmer. Herd-level factors associated with bulk tank SCC were determined using linear models with annual somatic cell score (i.e., arithmetic mean of the natural logarithm of bulk tank SCC) included as the dependent variable. All herd level factors were analysed individually in separate regression models, which included an adjustment for geographical location of the farm; a multiple regression model was subsequently developed. Management practices associated with low SCC included the use of dry cow therapy, participation in a milk recording scheme and the use of teat disinfection post-milking. There was an association between low SCC and an increased level of hygiene and frequency of cleaning of the holding yard, passageways and cubicles. Herd management factors associated with bulk tank SCC in Irish grazing herds are generally in agreement with most previous studies from confinement systems of milk production. PMID:22081962

  2. Association between somatic cell count and serial locomotion score assessments in UK dairy cows.

    PubMed

    Archer, S C; Green, M J; Madouasse, A; Huxley, J N

    2011-09-01

    This research investigated the effect of lameness, measured by locomotion score (LS) on the somatic cell count (SCC) of UK dairy cows. The data set consisted of 11,141 records of SCC and LS collected monthly on 12 occasions from 1,397 cows kept on 7 farms. The data were analyzed to account for the correlation of repeated measures of SCC within cow. Results were controlled for farm of origin, stage of lactation, parity, season, and test-day milk yield. Compared with the geometric mean SCC for cows with LS 1 on each farm, cows on farm 3 with LS 2 produced milk with 28,000 fewer somatic cells/mL, and cows with LS 2 on farm 6 produced milk with 30,000 fewer somatic cells/mL at a test day within 10 d. Cows that would have LS 3 six months later produced milk with 16,000 fewer somatic cells/mL compared with the geometric mean SCC for cows that would have LS 1 in 6 mo time. These results illustrate differences in disease dynamics between farms, highlight potential conflict between lameness and mastitis control measures, and emphasize the importance of developing farm-specific estimates of disease costs, and hence, health management plans in clinical practice. PMID:21854911

  3. Quantification of cell lysis during CHO bioprocesses: Impact on cell count, growth kinetics and productivity.

    PubMed

    Klein, Tobias; Heinzel, Nicole; Kroll, Paul; Brunner, Matthias; Herwig, Christoph; Neutsch, Lukas

    2015-08-10

    High cell densities and high viability are critical quality attributes for mammalian bioprocesses. Determination of living and dead cell numbers is nowadays routinely performed by automated image-based cell analyzers or flow cytometry. However, complete lysis of cells is usually neglected by these devices. We present a novel method for robust quantification of lysed cell populations over the course of a CHO bioprocess. The release of lactate dehydrogenase (LDH) and double stranded genomic DNA in culture supernatants were used as markers for cell lysis. We considered the degradation of both markers over cultivation time, which significantly increased the amount of released LDH and DNA. For correct and robust estimation of lysed cell fractions, degradation of both markers over cultivation time was considered, where redundancy of markers allowed data reconciliation. Calculating the number of cells which were subject to complete cell lysis, we could show that this fraction makes up as much as 30% of the total produced biomass and is not described by measurements of image-based analyzers. Finally, we demonstrate that disregarding cell lysis heavily affects the calculation of biomass yields and growth rates and that increasing levels of cell lysis are related to decreased productivity. PMID:25956245

  4. Measuring High-Order Moments of the Galaxy Distribution from Counts in Cells -- The Edgeworth Approximation

    E-print Network

    Rita Seungjung Kim; Michael A. Strauss

    1997-09-24

    To probe the weakly non-linear regime, past the point where simple linear theory is sufficient to describe the statistics of the density distribution, we measure the skewness (S_3) and kurtosis (S_4) of the Count Probability Distribution Function (CPDF) of the IRAS 1.2 Jy sample obtained from counts in cells. These quantities are free parameters in a maximum likelihood fit of an Edgeworth expansion convolved with a Poissonian to the observed CPDF. This method, applicable on scales greater than 5 Mpc, is appreciably less sensitive to the tail of the distribution than are measurements of S_3 and S_4 from moments of the CPDF. We measure S_3 and S_4 to l~50 h^{-1} Mpc; the data are consistent with scale invariance, yielding averages of = 2.83 +/- 0.09, and = 6.89 +/- 0.68. These values are higher than those found using the moments method on the same data set, = 1.5 +/- 0.5 and = 4.4 +/- 3.7, due to lack of correction in the latter work for finite-volume effects. Unlike the moments method, our results are quite robust to the fact that IRAS galaxies are under-represented in cluster cores. We use N-body simulations to show that our method yields unbiased results.

  5. Cultured leptomeningeal cells secrete cerebrospinal fluid proteins.

    PubMed

    Ohe, Y; Ishikawa, K; Itoh, Z; Tatemoto, K

    1996-09-01

    To extrapolate the function of the leptomeninges, we examined the profile of the proteins secreted from the cultured leptomeningeal cells prepared from 1-2-day-old rats. In sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis of the medium conditioned with the cultured cells, 20-25 differentially distinctive protein bands were noted. Through several chromatographic procedures (Sephadex G-75, Mono Q, and 7C8-300), altogether 18 proteins were purified to homogeneity, and the partial amino acid sequence of each protein was determined. Homology search revealed that the major proteins included prostaglandin-D-synthase or beta-trace protein, insulin-like growth factor (IGF)-II, IGF-binding protein-2, apolipoprotein E, beta 2-microglobulin, cystatin C, transferrin, peptidyl-prolyl cis-trans isomerase or cyclophilin C, secreted protein acidic and rich in cysteine, ubiquitin, lysozyme C, extracellular superoxide dismutase, and collagen alpha-1 (III). Most of these proteins are known to be the major brain-derived protein constituents of CSF and are thought to play important roles in certain biological events in the brain. Considering the morphological features, the present findings suggest the importance of the leptomeninges as an origin of such proteins in CSF. PMID:8752101

  6. Under-filled blood collection tubes containing K2EDTA as anticoagulant are acceptable for automated complete blood counts, white blood cell differential, and reticulocyte count.

    PubMed

    Xu, M; Robbe, V A; Jack, R M; Rutledge, J C

    2010-10-01

    Current laboratory standards from Clinical Laboratory Standards Institute (CLSI) and manufacturer's (Becton Dickinson) data indicate that under-filling K(2)EDTA blood collection tubes can result in erroneous hematology values. To accommodate under-filled tubes and reduce collection volumes while optimizing our automation, we explored the acceptable limit of under-filled tubes for hematology values. We collected 8.0 ml of blood from 30 normal adult volunteers. Each donation was aliquoted in the following volumes: 4.0, 2.0, 1.0, 0.5 ml x 2. These samples were analyzed within 1 h of blood collection on Sysmex XE-2100 (Sysmex America Inc., Mundelein, IL, USA) for complete blood count, reticulocyte, and white blood cell differentials. Results of the under-filled tubes were compared to those of the standard volume. The Deming regression analysis show excellent correlation for all parameters between each under-filled blood collection volume compared to a standard 4 ml volume. The Bland and Altman analysis shows good agreement between both 1.0 and 2.0 ml compared to a 4.0 ml volume. The 0.5 ml compared to a 4.0 ml volume, however, shows increased variation on many parameters. In addition all three collection volumes show negative bias compared to the standard volume for platelet count, but the difference is considered insignificant with a percent difference of 5.5%, 3.2%, and 1.5% for 0.5, 1.0, and 2.0 ml collection volume respectively. Finally for 0.5 ml collection volume we noticed a low level of false positive flagging rate for white blood cell. Acceptable complete blood count values of under-filled powdered K(2)EDTA tubes can be obtained with as little as 1.0 ml of blood. PMID:20041968

  7. Somatic cell count dynamics in a large sample of dairy herds in England and Wales.

    PubMed

    Madouasse, A; Huxley, J N; Browne, W J; Bradley, A J; Green, M J

    2010-08-01

    An essential reason to record and evaluate patterns of cow somatic cell count (SCC) within a dairy herd is to help in making clinical decisions on the control of mastitis. An understanding of when new infections occur and how patterns of infection influence herd bulk milk somatic cell count (BMSCC) are critical when implementing mastitis control because it enables advisors to target specific problem areas. The objective of this research was to evaluate individual cow SCC patterns in terms of their contribution to BMSCC. Data collected in 2128 herds from England and Wales between 2004 and 2006 were used. Cows were categorised as having a low, medium or high SCC based on thresholds of 100,000 cells/mL and 200,000 cells/mL. Movements between these categories in consecutive months, before or after 30 days in milk, in primiparous (heifers) and multiparous cows (cows) were used to predict BMSCC. From these categories, new variables representing different SCC patterns, were calculated and included in different models: the medium SCC category was grouped with either the low or the high category, and the denominator was either the total number of cows recorded during the herd-year or the number of cows eligible for a particular transition. Model fitting and predictions were carried out in a Bayesian framework. A random sample of 1500 herds was used for parameter estimation and the remaining 628 herds for model validation. Heifers were more likely to remain at, or to move to, a low SCC than cows. A transition threshold of 100,000 cells/mL for heifers resulted in a poorer model fit and predictive ability than a threshold of 200,000 cells/mL. A model using a single threshold of 200,000 cells/mL regardless of parity was the best to predict BMSCC. The sensitivity and specificity of this final model to correctly predict a BMSCC > 200, 000 cells/mL in the validation dataset were 86.5% and 86.8%, respectively. Important SCC patterns that influenced BMSCC were cows and heifers staying above 200,000 cells/mL for two consecutive recordings during lactation, cows moving from below to above 200,000 cells/mL across the dry period, cows remaining above 200,000 cells/mL across the dry period and heifers calving with an SCC above 200,000 cells/mL in the first month of lactation. The variation between herds in SCC transitions was evaluated and it was concluded that the performance of the top 10% of herds would be useful to provide benchmarks to evaluate dairy herd mastitis. PMID:20627342

  8. Possible Prognostic and Therapeutic Significance of c-Kit Expression, Mast Cell Count and Microvessel Density in Renal Cell Carcinoma

    PubMed Central

    Marech, Ilaria; Gadaleta, Cosmo Damiano; Ranieri, Girolamo

    2014-01-01

    Renal cell carcinoma (RCC) is the most frequent renal tumor and its incidence is increasing worldwide. Tumor angiogenesis is known to play a crucial role in the etiopathogenesis of RCC and over the last few years an even deeper knowledge of its contribution in metastatic RCC development has led to the development of numerous molecular targeting agents (such as sunitinib, sorafenib, pazopanib, axitinib, tivozanib, and dovitinib). The above agents are principally directed against vascular endothelial growth factor receptor (VEGFR) members and also against c-Kit receptor (c-KitR). The role of c-kitR inhibition on clear cell RCC (ccRCC), the main RCC subtype, is less well established. Whether c-kitR activation through its ligand, stem cell factor (SCF) contributes significantly to the effects of tyrosine kinase inhibitors (TKIs) treatment remains to be established. It is important to underscore that the c-KitR is expressed on mast cells (MCs) and cancer cells. After an examination of the c-KitR/SCF pathway, we review here the principal studies that have evaluated c-Kit expression in RCC. Moreover, we summarize some investigations that have observed the distribution of MCs in primary renal cancer and in adjacent normal tissue with appropriate histological immunohistochemical techniques. We also focus on few studies that have evaluated the correlation between RCC proliferation, MC count and microvessel density (MVD), as hallmarks of tumor angiogenesis. Thus, the aim of this review of the literature is to clarify if c-KitR expression, MC count and MVD could have prognostic significance and the possible predictive therapeutic implications in RCC. PMID:25056544

  9. Possible prognostic and therapeutic significance of c-Kit expression, mast cell count and microvessel density in renal cell carcinoma.

    PubMed

    Marech, Ilaria; Gadaleta, Cosmo Damiano; Ranieri, Girolamo

    2014-01-01

    Renal cell carcinoma (RCC) is the most frequent renal tumor and its incidence is increasing worldwide. Tumor angiogenesis is known to play a crucial role in the etiopathogenesis of RCC and over the last few years an even deeper knowledge of its contribution in metastatic RCC development has led to the development of numerous molecular targeting agents (such as sunitinib, sorafenib, pazopanib, axitinib, tivozanib, and dovitinib). The above agents are principally directed against vascular endothelial growth factor receptor (VEGFR) members and also against c-Kit receptor (c-KitR). The role of c-kitR inhibition on clear cell RCC (ccRCC), the main RCC subtype, is less well established. Whether c-kitR activation through its ligand, stem cell factor (SCF) contributes significantly to the effects of tyrosine kinase inhibitors (TKIs) treatment remains to be established. It is important to underscore that the c-KitR is expressed on mast cells (MCs) and cancer cells. After an examination of the c-KitR/SCF pathway, we review here the principal studies that have evaluated c-Kit expression in RCC. Moreover, we summarize some investigations that have observed the distribution of MCs in primary renal cancer and in adjacent normal tissue with appropriate histological immunohistochemical techniques. We also focus on few studies that have evaluated the correlation between RCC proliferation, MC count and microvessel density (MVD), as hallmarks of tumor angiogenesis. Thus, the aim of this review of the literature is to clarify if c-KitR expression, MC count and MVD could have prognostic significance and the possible predictive therapeutic implications in RCC. PMID:25056544

  10. MBL or CLL: Which classification best categorizes the clinical course of patients with an absolute lymphocyte count ? 5 × 10 9 L ?1 but a B-cell lymphocyte count < 5 × 10 9 L ?1?

    Microsoft Academic Search

    Tait D. Shanafelt; Neil E. Kay; Tim G. Call; Clive S. Zent; Diane F. Jelinek; Betsy LaPlant; William G. Morice; Curtis A. Hanson

    2008-01-01

    To eliminate overlap with monoclonal B-cell lymphocytosis (MBL), some have proposed basing the diagnosis of chronic lymphocytic leukemia (CLL) on B lymphocyte count rather than absolute lymphocyte count (ALC). Such criteria should be based, in part, on patient outcomes. We evaluated the clinical implications of the proposed re-classification in 112 consecutive, newly diagnosed, Rai stage 0 patients. The new criteria

  11. Red blood cell count as an indicator of microvascular complications in Chinese patients with type 2 diabetes mellitus

    PubMed Central

    Wang, Zhan-Sheng; Song, Zhan-Chun; Bai, Jing-Hui; Li, Fei; Wu, Tao; Qi, Ji; Hu, Jian

    2013-01-01

    Background Rheological disorders of red blood cells (RBC) and decreased RBC deformability have been involved in the development of diabetic microangiopathy. However, few studies have evaluated the association of RBC count with microvascular complications in patients with type 2 diabetes mellitus (T2DM). The purpose of this study was to investigate the association of RBC count with microvascular complications in patients with T2DM. Methods This study involved 369 patients with T2DM: 243 with one or more microvascular complications and 126 without microvascular complications. Anticoagulated blood was collected and analyzed in an automated blood cell counter. The presence of risk factors for microvascular complications was determined. Results The proportion of patients with microvascular complications increased as the RBC count decreased (P < 0.001). After adjustment for known risk factors for microvascular complications by logistic regression analysis, lower quartiles of RBC count were associated with a higher risk of microvascular complications compared with the reference group composed of the highest quartile (first quartile, odds ratio 4.98, 95% confidence interval 1.54–6.19, P = 0.008; second quartile, odds ratio 3.21, 95% confidence interval 1.17–5.28, P = 0.024). Conclusion A decreased RBC count is associated with microvascular complications in Chinese patients with T2DM. The RBC count is a potential marker to improve further the ability to identify diabetic patients at high risk of microvascular complications. PMID:23690689

  12. Effect of exercise on erythrocyte count and blood activity concentration after technetium-99m in vivo red blood cell labeling

    SciTech Connect

    Konstom, M.A. (New England Medical Center, Boston, MA); Tu'meh, S.; Wynne, J.; Beck, J.R.; Kozlowski, J.; Holman, B.L.

    1982-09-01

    The effects of exercise on blood radiotracer concentration after technetium-99m in vivo red blood cell labeling was studied. After red blood cell labeling, 13 subjects underwent maximal supine bicycle exercise. Radioactivity, analyzed with a well counter, was measured in heparinized venous blood samples drawn at rest and during peak exercise. Changes in activity were compared with changes in erythrocyte count. Activity and erythrocyte counts increased in erythrocyte count (r=0.78), but did not correlate with either duration of exercise or maximal heart rate. Twenty minutes after termination of exercise, activity and erythrocyte count had decreased from peak exercise values but remained higher than preexercise values. In nine nonexercised control subjects, samples drawn 20 minutes apart showed no change in activity or in erythrocyte count. It was concluded that exercise increases blood activity, primarily because of an increase in erythrocyte count. During radionuclide ventriculography, blood activity must be measured before and after any intervention, particularly exercise, before a change in left ventricular activity can be attributed to a change in left ventricular volume.

  13. Prevalence of isosporiasis in relation to CD4 cell counts among HIV-infected patients with diarrhea in Odisha, India

    PubMed Central

    Mohanty, Indrani; Panda, Pritilata; Sahu, Susmita; Dash, Mutikesh; Narasimham, Moningi Venkat; Padhi, Sanghamitra; Parida, Banojini

    2013-01-01

    Background: To determine the prevalence of Isospora belli and its correlation with CD4+ cell counts in HIV-positive patients with diarrhea in this region. Materials and Methods: Stool samples from 250 HIV-positive patients, including 200 with diarrhea and 50 without diarrhea included in the study were examined for the presence of enteric parasites under microscopy. Prevalence of the enteric parasites with special reference to I. belli in HIV-positive patients with and without diarrhea were calculated and correlated with their CD4+ cell counts. Results: Enteric parasites were detected in 39% of the HIV patients with diarrhea compared to 30% without diarrhea. I. belli was detected in 22% of the patients with diarrhea and in 4% without diarrhea (P = 0.0019). I. belli was the most common parasite, followed by Entamoeba histolytica/dispar (8%) and Cryptosporidium parvum (5%) in HIV-positive patients with diarrhea. In HIV-positive patients without diarrhea, the most common parasite detected was E. histolytica/dispar (12%) followed by C. parvum (6%) and I. belli (4%). The mean CD4 cell count of HIV-positive patients with diarrhea suffering from isosporiasis was 138.35 ± 70.71. In patients with CD4 cell counts <200/?l, I. belli was seen in 36/123 stool samples and 2/27 stool samples which was statistically significant (P = 0.0157). Conclusion: I. belli was the predominant parasite with a prevalence of 22% among HIV-positive patients with diarrhea, majority having CD4 cell count <200/?l. This study highlights the importance of routine screening for coccidian parasites in HIV-positive patients with and without diarrhea especially in those with low CD4 cell counts. PMID:24223376

  14. Association between somatic cell count after first parturition and cumulative milk yield in dairy cows.

    PubMed

    Archer, S C; Mc Coy, F; Wapenaar, W; Green, M J

    2013-10-01

    The aim was to assess the association between the somatic cell count of parity 1 cows between 5 and 30 days in milk (SCC1), and subsequent cumulative milk yield over approximately two years for cows in English and Welsh dairy herds. The dataset included records from 43,461 cows in 2111 herds, from 2004 to 2006. Cumulative milk yield was the model outcome, and a random effect was included to account for variation between herds. The model fitted the data well and was used to make predictions of cumulative milk yield, based on SCC1. A unit increase in the natural logarithm of SCC1/1000 was associated with a median decrease in cumulative milk yield of 482 kg, over a median study period of 868 days. PMID:23920363

  15. Parallel-plate fluid flow systems for bone cell stimulation.

    PubMed

    Huesa, Carmen; Helfrich, Miep H; Aspden, Richard M

    2010-04-19

    Bone responds to changes in its mechanical environment, but the mechanisms by which it does so are poorly understood. One hypothesis of mechanosensing in bone states that osteocytes can sense the flow of fluid through the canalicular system. To study this in vitro a number of fluid flow devices have been designed in which cells are placed between parallel plates in sealed chambers. Fluid flows through the chambers at controlled rates, most commonly driven by a peristaltic pump. In addition to fluid flow, high pressures have been observed in these chambers, but the effect of this on the cellular responses has generally been ignored or considered irrelevant, something challenged by recent cellular experiments using pressure only. We have, therefore, devised a system in which we can considerably reduce the pressure while maintaining the flow rate to enable study of their effects individually and in combination. As reducing pressure also reduces the risk of leaks in flow chambers, our system is suitable for real-time microscopical experiments. We present details of the new systems and of experiments with osteoblasts to illustrate the effects of fluid flow with and without additional pressure on the translocation of beta-catenin to the nucleus. PMID:20031135

  16. Immunophenotypic analysis of cerebrospinal fluid cell populations with the Cell-Dyn Sapphire haematology analyser: method feasibility and preliminary observations.

    PubMed

    Adam, P; Sobek, O; Scott, C S; Dolezil, D; Kasik, J; Hajdukova, L; Adam, D

    2010-02-01

    Cerebrospinal fluid (CSF) samples (n=50) from patients with neurological disease (bacterial infection, viral infection, neuroborreliosis and multiple sclerosis) were analysed to characterize cell populations by fluorescent immunocytometry with the CD-Sapphire haematology analyser. Reagent combinations applied to all CSF samples comprised CD3/CD19/HLA-DR and CD4/CD8, with some being further analysed using CD3/CD4, CD3/CD16 and CD3/CD25 protocols. Of the 50 samples, 11 were excluded because of high proportions of nonviable cells (n=2) or insufficient cell numbers (n=9). Apart from bacterial infection with granulocytosis, all diagnostic groups showed high proportions (51.4-77.0%) of CD3+ T cells. There was a modest association between T-cell and B-cell counts, but absolute B-cell numbers exceeded 5 cells/microl in only 7/39 cases (neuroborreliosis, n=6; bacterial meningitis, n=1). CD3/Ia antigen (activation) co-expression was low and only exceeded 5% in 7/39 samples with no diagnostic correlation. Primary CD4+ and CD8+ T-cell subsets showed similar quantitative trends and CD4/CD8 co-analysis revealed the presence in all diagnostic groups (neuroborreliosis and multiple sclerosis in particular) of a CD4+CD8int fraction that was predominantly CD3+ and CD16- and had a morphological profile consistent with small lymphoid cells. Supplementary CD-Sapphire cellular immunological analysis of most CSF samples is feasible using the procedure detailed in this communication. PMID:19500178

  17. Each cell counts: Hematopoiesis and immunity research in the era of single cell genomics.

    PubMed

    Jaitin, Diego Adhemar; Keren-Shaul, Hadas; Elefant, Naama; Amit, Ido

    2015-02-01

    Hematopoiesis and immunity are mediated through complex interactions between multiple cell types and states. This complexity is currently addressed following a reductionist approach of characterizing cell types by a small number of cell surface molecular features and gross functions. While the introduction of global transcriptional profiling technologies enabled a more comprehensive view, heterogeneity within sampled populations remained unaddressed, obscuring the true picture of hematopoiesis and immune system function. A critical mass of technological advances in molecular biology and genomics has enabled genome-wide measurements of single cells - the fundamental unit of immunity. These new advances are expected to boost detection of less frequent cell types and fuzzy intermediate cell states, greatly expanding the resolution of current available classifications. This new era of single-cell genomics in immunology research holds great promise for further understanding of the mechanisms and circuits regulating hematopoiesis and immunity in both health and disease. In the near future, the accuracy of single-cell genomics will ultimately enable precise diagnostics and treatment of multiple hematopoietic and immune related diseases. PMID:25727184

  18. Cell detection and counting through cell lysate impedance spectroscopy in microfluidic devices{

    E-print Network

    Demirci, Utkan

    -fluidic and quantifying cell number, can serve as a single use cartridge for a hand-held instrument to provide simple approach can be easily adapted to a wide spectrum of clinical applications, detecting these isolated cells remains a technical challenge to be addressed. The use of optical microscopy for detection and quantifica

  19. Fluid and Cell Transport Through a Microfabricated Flow Chamber.

    NASA Astrophysics Data System (ADS)

    Brody, James Patrick

    We use silicon processing techniques to construct microfabricated fluid flow chambers. Custom designed silicon wafers with feature sizes of 1-10 ?m and etch depths from 0.5-5 ?m are anodically bonded to Pyrex glass to create a hermetically sealed chamber. A pressure gradient is placed across the chamber to induce bulk fluid flow. Properties of fluid flow and red blood cells are recorded using video microscopy. The human red blood cell is ideal for studying cellular membranes. It is an 8 ?m diameter biconcave disc containing a membrane and associated cytoskeleton which surrounds a thick solution of hemoglobin. The material properties of individual red blood cells have been extensively studied in the past using micropipettes. However, we can get statistics on hundreds of red blood cells by fabricating an array of narrow channels 4 mu m x 4 ?m in cross-section (the diameter of the smallest capillaries in the human body) and 13 ?m long. These narrow channels are followed by an open space. This geometry forces red cells to repeatedly fold and unfold. Using these arrays, we show that the shear modulus of the membrane does not have a unique value, but has a distribution that ranges from 3-12 times 10 ^{-6} N/m. The surprisingly wide distribution is not due to cell size or cell age. It does seem to be correlated with intracellular Ca^ {2+}<=vels, leading us to believe that cell rigidity is controlled by some active process. We also report observations on red blood cells changing their rigidity by factors of fifty over tens of seconds. These microfabricated flow chambers are ideal for studying fluid flow through porous media. We construct custom designed two-dimensional environments with micron size features. These environments can be described by simple analytical theories which also attempt to describe flow through rock. For example, we image viscous imbibition of water into a percolation grid with 5 mu m edges in real time, and measure the permeability as a function of concentration for a simple rectangular array geometry.

  20. Efficacy of optimized in vitro predegeneration period on the cell count and purity of canine Schwann cell cultures

    PubMed Central

    Niapour, Nazila; Mohammadi-Ghalehbin, Behnam; Golmohammadi, Mohammad Ghasem; Amani, Mohammad; Salehi, Hossein; Niapour, Ali

    2015-01-01

    Objective(s): Predegeneration is a standard technique to obtain mitotically activated and enriched cultures of Schwann cells (SCs). This study, for the first time, evaluated the impact of various duration of predegeneration on cell yield and enrichment of SCs from dog peripheral nerve. Materials and Methods: Dog sural nerves were subjected to 5, 10, 15 day-long in vitro predegeneration. The total cell yield and the purity of SCs were evaluated in each group on the first and seventh day after plating. Results: The maximum and minimum numbers of cells were counted in 15 day-long predegene-ration and control groups which underwent no predegeneration. The 10 day-long in vitro predegeneration group with 80±0.5% SCs enrichment had the best purity after plating day and could maintain its purity with elapsing on cultures. Conclusion: 10 day-long predegeneration results in the higher cell number and the better and prolonged purity of SCs in culture. PMID:25945245

  1. White Blood Cell Count Measured Prior to Cancer Development Is Associated with Future Risk of Venous Thromboembolism – The Tromsø Study

    PubMed Central

    Blix, Kristine; Jensvoll, Hilde; Brækkan, Sigrid K.; Hansen, John-Bjarne

    2013-01-01

    Background Elevated white blood cell (WBC) count is associated with risk of venous thromboembolism (VTE) in cancer patients initiating chemotherapy. It is not known whether the risk of VTE by WBC count in cancer patients is causal or merely a consequence of the malignant disease. To address this question, we studied the association between WBC count, measured prior to cancer development, and risk of VTE in subjects who did and did not develop cancer during follow-up in a prospective population-based study. Methods Baseline characteristics, including WBC and neutrophil counts, were measured in 24304 initially cancer-free subjects who participated in the Tromsø Study in 1994-1995. Incident cancer diagnosis and VTE events were registered up to September 1, 2007. In the cancer cohort, WBC and neutrophil counts were measured in average 7.1 years before cancer development. Cox-regression models were used to calculate hazard ratios (HRs) for VTE by WBC and neutrophil counts as categorized variables (<40th, 40-80th, and >80th percentile) with 95% confidence intervals (CIs). Results During follow-up, 1720 subjects developed cancer and there were 388 VTE events, of which 116 occurred in the cancer-group (6.9 per 1000 person-years) and 272 in the cancer-free group (1.1 per 1000 person-years). In those who developed cancer, WBC count above the 80th percentile (?8.6x109 cells/L) was associated with a 2.4-fold higher risk (HR 2.36, 95% CI: 1.44-3.87) of VTE compared to WBC count below the 40th percentile (<6.4x109 cells/L). No association was found between WBC count and VTE in those who stayed cancer-free (HR 0.94, 95% CI 0.65-1.36). Similar findings were observed for neutrophils. Comment Pre-cancer WBC count was associated with risk of VTE in cancer patients, but not in cancer-free subjects. Our findings suggest that leukocytes may play a causal role in cancer-related VTE rather than only reflecting the low-grade inflammation associated with cancer. PMID:24023876

  2. Prolonged effect of fluid flow stress on the proliferative activity of mesothelial cells after abrupt discontinuation of fluid streaming

    SciTech Connect

    Aoki, Shigehisa, E-mail: aokis@cc.saga-u.ac.jp [Department of Pathology and Microbiology, Faculty of Medicine, Saga University, Saga (Japan)] [Department of Pathology and Microbiology, Faculty of Medicine, Saga University, Saga (Japan); Ikeda, Satoshi [Department of Pathology and Microbiology, Faculty of Medicine, Saga University, Saga (Japan)] [Department of Pathology and Microbiology, Faculty of Medicine, Saga University, Saga (Japan); Takezawa, Toshiaki [Transgenic Animal Research Center, National Institute of Agrobiological Sciences, Ibaraki (Japan)] [Transgenic Animal Research Center, National Institute of Agrobiological Sciences, Ibaraki (Japan); Kishi, Tomoya [Department of Internal Medicine, Saga University, Saga (Japan)] [Department of Internal Medicine, Saga University, Saga (Japan); Makino, Junichi [Makino Clinic, Saga (Japan)] [Makino Clinic, Saga (Japan); Uchihashi, Kazuyoshi; Matsunobu, Aki [Department of Pathology and Microbiology, Faculty of Medicine, Saga University, Saga (Japan)] [Department of Pathology and Microbiology, Faculty of Medicine, Saga University, Saga (Japan); Noguchi, Mitsuru [Department of Urology, Faculty of Medicine, Saga University, Saga (Japan)] [Department of Urology, Faculty of Medicine, Saga University, Saga (Japan); Sugihara, Hajime [Department of Physical Therapy, International University of Health and Welfare, Fukuoka (Japan)] [Department of Physical Therapy, International University of Health and Welfare, Fukuoka (Japan); Toda, Shuji [Department of Pathology and Microbiology, Faculty of Medicine, Saga University, Saga (Japan)] [Department of Pathology and Microbiology, Faculty of Medicine, Saga University, Saga (Japan)

    2011-12-16

    Highlights: Black-Right-Pointing-Pointer Late-onset peritoneal fibrosis leading to EPS remains to be elucidated. Black-Right-Pointing-Pointer Fluid streaming is a potent factor for peritoneal fibrosis in PD. Black-Right-Pointing-Pointer We focused on the prolonged effect of fluid streaming on mesothelial cell kinetics. Black-Right-Pointing-Pointer A history of fluid streaming exposure promoted mesothelial proliferative activity. Black-Right-Pointing-Pointer We have thus identified a potent new factor for late-onset peritoneal fibrosis. -- Abstract: Encapsulating peritoneal sclerosis (EPS) often develops after transfer to hemodialysis and transplantation. Both termination of peritoneal dialysis (PD) and transplantation-related factors are risks implicated in post-PD development of EPS, but the precise mechanism of this late-onset peritoneal fibrosis remains to be elucidated. We previously demonstrated that fluid flow stress induced mesothelial proliferation and epithelial-mesenchymal transition via mitogen-activated protein kinase (MAPK) signaling. Therefore, we speculated that the prolonged bioactive effect of fluid flow stress may affect mesothelial cell kinetics after cessation of fluid streaming. To investigate how long mesothelial cells stay under the bioactive effect brought on by fluid flow stress after removal of the stress, we initially cultured mesothelial cells under fluid flow stress and then cultured the cells under static conditions. Mesothelial cells exposed to fluid flow stress for a certain time showed significantly high proliferative activity compared with static conditions after stoppage of fluid streaming. The expression levels of protein phosphatase 2A, which dephosphorylates MAPK, in mesothelial cells changed with time and showed a biphasic pattern that was dependent on the duration of exposure to fluid flow stress. There were no differences in the fluid flow stress-related bioactive effects on mesothelial cells once a certain time had passed. The present findings show that fluid flow stress exerts a prolonged bioactive effect on mesothelial cells after termination of fluid streaming. These findings support the hypothesis that a history of PD for a certain period could serve as a trigger of EPS after stoppage of PD.

  3. Influence of storage and preservation on fossomatic cell count and composition of goat milk.

    PubMed

    Sánchez, A; Sierra, D; Luengo, C; Corrales, J C; Morales, C T; Contreras, A; Gonzalo, C

    2005-09-01

    This study was designed to evaluate the effects of different test conditions on the somatic cell count (SCC) and composition of goat milk. To this end, 3600 tests were performed on 1800 aliquots taken from 40 goat milk samples using a combined instrument set-up based on flow cytometry for SCC and Fourier transform infrared analysis for fat, total protein, lactose, total solids, and freezing point determinations. The conditions tested were storage temperature (refrigeration and freezing), use of a preservative [no preservative (NP), azidiol (AZ), and bronopol (BR)], and age of the milk samples at each storage temperature (24 h to 42 d at refrigeration temperature and 21 to 105 d at freezing temperature). Significant effects on logSCC variation were shown by the storage temperature, the preservation treatment, the interaction of storage temperature x preservation treatment, and milk age within the interaction of storage temperature x preservative. Highest counts were recorded in the BR-preserved milk samples (logSCC = 5.877), and lowest counts were recorded in milk samples preserved using AZ (logSCC = 5.803). The use of frozen/thawed samples led to a significantly decreased logSCC for the treatments AZ and NP; the logSCC was not modified when BR-preserved frozen/thawed samples were analyzed. During storage, variations in the SCC observed for BR-preserved samples stored at refrigeration temperature for up to 25 d and at freezing temperature for all times tested were always < 10%. The preservation treatment was the main factor affecting the milk composition variables examined. Highest values of most variables were obtained in the BR-preserved samples, and the lowest values were obtained in the AZ-preserved samples. The freezing point was lower in the preserved samples than in the NP samples. The levels of milk constituents recorded in the BR-preserved samples were independent of both the storage temperature and age of milk sample. Our findings indicate that the freezing point of goat milk must be interpreted according to the preservative used. PMID:16107398

  4. Cell sourcing for bone tissue engineering: Amniotic fluid stem cells have a delayed, robust differentiation compared to mesenchymal stem cells

    Microsoft Academic Search

    Alexandra Peister; Maria A. Woodruff; Jarod J. Prince; Derwin P. Gray; Dietmar W. Hutmacher; Robert E. Guldberg

    2011-01-01

    Cell based therapies for bone regeneration are an exciting emerging technology, but the availability of osteogenic cells is limited and an ideal cell source has not been identified. Amniotic fluid-derived stem cells (AFS) and bone-marrow derived mesenchymal stem cells (MSCs) were compared to determine their osteogenic differentiation capacity in both 2D and 3D environments. In 2D culture, the AFS cells

  5. Three-dimensional counting of morphologically normal human red blood cells via digital holographic microscopy

    NASA Astrophysics Data System (ADS)

    Yi, Faliu; Moon, Inkyu; Lee, Yeon H.

    2015-01-01

    Counting morphologically normal cells in human red blood cells (RBCs) is extremely beneficial in the health care field. We propose a three-dimensional (3-D) classification method of automatically determining the morphologically normal RBCs in the phase image of multiple human RBCs that are obtained by off-axis digital holographic microscopy (DHM). The RBC holograms are first recorded by DHM, and then the phase images of multiple RBCs are reconstructed by a computational numerical algorithm. To design the classifier, the three typical RBC shapes, which are stomatocyte, discocyte, and echinocyte, are used for training and testing. Nonmain or abnormal RBC shapes different from the three normal shapes are defined as the fourth category. Ten features, including projected surface area, average phase value, mean corpuscular hemoglobin, perimeter, mean corpuscular hemoglobin surface density, circularity, mean phase of center part, sphericity coefficient, elongation, and pallor, are extracted from each RBC after segmenting the reconstructed phase images by using a watershed transform algorithm. Moreover, four additional properties, such as projected surface area, perimeter, average phase value, and elongation, are measured from the inner part of each cell, which can give significant information beyond the previous 10 features for the separation of the RBC groups; these are verified in the experiment by the statistical method of Hotelling's T-square test. We also apply the principal component analysis algorithm to reduce the dimension number of variables and establish the Gaussian mixture densities using the projected data with the first eight principal components. Consequently, the Gaussian mixtures are used to design the discriminant functions based on Bayesian decision theory. To improve the performance of the Bayes classifier and the accuracy of estimation of its error rate, the leaving-one-out technique is applied. Experimental results show that the proposed method can yield good results for calculating the percentage of each typical normal RBC shape in a reconstructed phase image of multiple RBCs that will be favorable to the analysis of RBC-related diseases. In addition, we show that the discrimination performance for the counting of normal shapes of RBCs can be improved by using 3-D features of an RBC.

  6. Microvessel count predicts metastatic disease and survival in non-small cell lung cancer.

    PubMed

    Fontanini, G; Bigini, D; Vignati, S; Basolo, F; Mussi, A; Lucchi, M; Chine, S; Angeletti, C A; Harris, A L; Bevilacqua, G

    1995-09-01

    The growth of newly formed vessels, or neoangiogenesis, represents an important step in both physiological and pathological situations: in particular, tumour growth and metastasis require angiogenesis. Microvessel count (MC), which represents a measure of tumour angiogenesis, has been associated with metastatic spread in cutaneous, mammary, prostatic, head and neck, and early-stage lung cancer. In this study, the role of tumour angiogenesis as a prognostic indicator was examined in 253 primary non-small lung cancer (NSCLC) patients. Microvessels were counted by highlighting endothelial cells with anti-Factor VIII monoclonal antibody (Mab) in methacarn-fixed tumour samples. In univariat analysis, MC (P< 0.000001), sex (P=0.0036), histotype (P < 0.014), tumour status (P <0.007), and vessel invasion (P < 0.019) were significantly related to hilar and/or mediastinal nodal involvement. However, in the stepwise logistic regression analysis, MC (P<0.000003) retained the most important influence on nodal metastasis. The overall survival analysis calculated by the Kaplan-Meier method revealed that tumours with high MC ( > 25 vessels/field) were significantly associated with increased death risk (log-rank test P = 0.00067; Cox's test P = 0.00046; Gehan's Wilcoxon test P = 0.00108). In 94 patients, the development of metastatic disease during follow-up was significantly related to MC. Indeed, patients who developed metastasis during follow-up showed a higher MC, either as a dichotomous (P = 0.01) or as a continuous (P = 0.003) variable, than patients who had developed no metastasis at the time of the analysis. Moreover, in the stepwise logistic regression analysis, MC retained the most important influence on distant metastases.(ABSTRACT TRUNCATED AT 250 WORDS) PMID:7472781

  7. Relation of admission white blood cell count to left ventricular remodeling after anterior wall acute myocardial infarction.

    PubMed

    Bauters, Anne; Ennezat, Pierre V; Tricot, Olivier; Lallemant, Robert; Aumégeat, Valérie; Segrestin, Benoit; Quandalle, Philippe; Lamblin, Nicolas; Bauters, Christophe

    2007-07-15

    We investigated whether a high white blood cell (WBC) count on admission for acute myocardial infarction (AMI) may be associated with a higher risk of subsequent left ventricular (LV) remodeling. We included 107 patients with anterior AMI. Echocardiographic studies were performed at hospital discharge, at 3 months, and at 1 year after AMI. LV remodeling (>20% increase in end-diastolic volume) was observed in 27% of patients. WBC counts during hospitalization were higher in patients who subsequently underwent LV remodeling (p = 0.003 for WBC count on admission). The increase in end-diastolic volume from baseline to 1 year was greater for patients in the higher tertile of WBC count on admission (p = 0.04). When adjusting for baseline clinical and echocardiographic characteristics by multivariate analysis, WBC count on admission was independently associated with LV remodeling (odds ratio 1.23, 95% confidence interval 1.04 to 1.45, p = 0.018). In conclusion, a high WBC count on admission for AMI is an independent predictor of LV remodeling, even when predischarge echocardiographic variables are taken into account. PMID:17631066

  8. Influence of Calicophoron microbothrium amphistomosis on the biochemical and blood cell counts of cattle.

    PubMed

    Mavenyengwa, M; Mukaratirwa, S; Monrad, J

    2010-12-01

    Sixteen Tuli steers aged 1 year were subdivided into four equal groups (I-IV) and infected with Calicophoron microbothrium metacercariae. Group I received a low dose (LD) of 5000 metacercariae, group II a medium dose (MD) of 15,000 metacercariae, group III a high dose (HD) of 25,000 metacercariae while group IV was the non-infected control (C) group. The experimental animals were monitored daily for clinical signs while ethylene diamine tetraacetic acid (EDTA) blood and serum samples were collected every 7 days until day 28 post-infection, when sample collection was terminated. Samples were processed for full blood count, eosinophils and blood biochemical values for calcium, magnesium, phosphorus, total protein and albumin. Moderate to severe diarrhoea developed in the MD and HD groups at day 21 post-infection. The diarrhoea coincided with a significant decrease (P < 0.05) in total plasma protein, calcium and phosphorus levels, particularly in the MD group. Similarly, a significant decrease (P < 0.05) in the packed cell volume (PCV), the haemoglobin (Hb) and red blood cell (RBC) levels occurred in the MD and HD groups from day 21 post-infection, while a significant increase (P < 0.05) in the circulating eosinophils occurred between 7 and 21 days post-infection in the LD and the HD groups. PMID:20109245

  9. The use of the white cell count and haemoglobin in combination as an effective screen to predict the normality of the full blood count

    PubMed Central

    OSEI-BIMPONG, A; McLEAN, R; BHONDA, E; LEWIS, S M

    2012-01-01

    Introduction The utility of the full blood count (FBC) is vast with each parameter serving as a tool to aid diagnosis and monitor disease progression. However, the effectiveness of the test is hampered because of increased workload and lack of interpretation. In the effort to redress this issue, the combined use of the white blood cell count (WBC) and haemoglobin in predicting the normality of the FBC is evaluated. Method FBC data were collated from 2191 patients and classified into two groups depending on whether the WBC and the haemoglobin were within the reference range. Blood films were examined on the abnormal FBC samples in each group and graded on morphology. Results The FBC was normal in 89.6% of cases in the presence of a normal WBC and haemoglobin with subtle abnormalities in the remainder; 1+ grading of abnormal morphology in 93%. However, when the WBC and/or haemoglobin was abnormal, the remaining FBC was significantly abnormal (P < 0.05) and the corresponding blood films were grossly abnormal; 2+/3+ grading in 96% of cases. Conclusion We concluded that in the presence of a normal WBC and haemoglobin, the FBC is normal in almost all cases and measuring these two parameters could be used as an effective screen to predict FBC normality. PMID:21883968

  10. Consequence of alternative standards for bulk tank somatic cell count of dairy herds in the United States

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Comparison of dairy operations failing compliance with current US and European Union (EU) standards for bulk-tank somatic cell count (BTSCC) as well as BTSCC standards proposed by 3 national organizations were evaluated using 2 populations of US dairy herds: Dairy Herd Improvement Association (DHI) ...

  11. Modeling HIV-1 Dynamics and the Effects of Decreasing Activated Infected T-cell Count by Filtration

    E-print Network

    Stryker, Gabrielle A.

    Modeling HIV-1 Dynamics and the Effects of Decreasing Activated Infected T-cell Count by Filtration. In this paper a model is developed that characterizes the rate of filtrations impact on disease progression of on modeling HIV-1 treatment as a filtration process where disease progression is studied as a function

  12. Influence of Estrus of Dairy Goats on Somatic Cell Count, Milk Traits, and Sex Steroid Receptors in the Mammary Gland

    Microsoft Academic Search

    P. Moroni; G. Pisoni; G. Savoini; E. van Lier; S. Acuña; J. P. Damián; A. Meikle

    2007-01-01

    Two experiments were conducted to study the effect of the stage of a spontaneous estrus cycle on milk yield and constituents (somatic cell count (SCC), fat, protein, caseins, lactose, and urea content) and on estrogen re- ceptor-? (ER?) and progesterone receptor (PR) immu- nostaining in the mammary gland. In experiment I, the major components of milk and SCC were monitored

  13. The influence of the season on the chemical composition and the somatic cell count of bulk tank cow's milk

    Microsoft Academic Search

    Bela Njari; Marko Samardžija

    The aim of our research was to establish the influence of the season on the chemical composition of bulk tank cow's milk and the somatic cell count in it. The bulk tank cow's milk samples were collected daily during a period of one year. When the milk was brought to the dairy in special tanks, before pouring bulk tank milk,

  14. Effects of stage of lactation, production, parity and season on somatic cell counts in infected and uninfected dairy goats

    Microsoft Academic Search

    David J. Wilson; Keith N. Stewart; Philip M. Sears

    1995-01-01

    A commercial dairy goat farm with 380 Alpine milking does maintained monthly records of milk production and somatic cell counts (SCC). Composite milk samples from all lactating does were cultured for mastitis pathogens every 6 months. Dairy Herd Improvement Association records were combined with culture results. Goats' SCC increased with intramammary infection (IMI). Increasing stage of lactation was also associated

  15. Management Style and Its Association with Bulk Milk Somatic Cell Count and Incidence Rate of Clinical Mastitis

    Microsoft Academic Search

    H. W. Barkema; J. D. Van der Ploeg; Y. H. Schukken; T. J. G. M. Lam; G. Benedictus; A. Brand

    1999-01-01

    Management style and its association with bulk milk somatic cell count (SCC) and the incidence rate of clinical mastitis were studied in 300 Dutch dairy herds. Cluster analysis was used to identify groups of farmers who had similar management styles for the prevention of mastitis. Two groups of farmers could be differentiated. The management style of the first group of

  16. Fluid models and simulations of biological cell phenomena

    NASA Technical Reports Server (NTRS)

    Greenspan, H. P.

    1982-01-01

    The dynamics of coated droplets are examined within the context of biofluids. Of specific interest is the manner in which the shape of a droplet, the motion within it as well as that of aggregates of droplets can be controlled by the modulation of surface properties and the extent to which such fluid phenomena are an intrinsic part of cellular processes. From the standpoint of biology, an objective is to elucidate some of the general dynamical features that affect the disposition of an entire cell, cell colonies and tissues. Conventionally averaged field variables of continuum mechanics are used to describe the overall global effects which result from the myriad of small scale molecular interactions. An attempt is made to establish cause and effect relationships from correct dynamical laws of motion rather than by what may have been unnecessary invocation of metabolic or life processes. Several topics are discussed where there are strong analogies droplets and cells including: encapsulated droplets/cell membranes; droplet shape/cell shape; adhesion and spread of a droplet/cell motility and adhesion; and oams and multiphase flows/cell aggregates and tissues. Evidence is presented to show that certain concepts of continuum theory such as suface tension, surface free energy, contact angle, bending moments, etc. are relevant and applicable to the study of cell biology.

  17. Automatic choroid cells segmentation and counting based on approximate convexity and concavity of chain code in fluorescence microscopic image

    NASA Astrophysics Data System (ADS)

    Lu, Weihua; Chen, Xinjian; Zhu, Weifang; Yang, Lei; Cao, Zhaoyuan; Chen, Haoyu

    2015-03-01

    In this paper, we proposed a method based on the Freeman chain code to segment and count rhesus choroid-retinal vascular endothelial cells (RF/6A) automatically for fluorescence microscopy images. The proposed method consists of four main steps. First, a threshold filter and morphological transform were applied to reduce the noise. Second, the boundary information was used to generate the Freeman chain codes. Third, the concave points were found based on the relationship between the difference of the chain code and the curvature. Finally, cells segmentation and counting were completed based on the characteristics of the number of the concave points, the area and shape of the cells. The proposed method was tested on 100 fluorescence microscopic cell images, and the average true positive rate (TPR) is 98.13% and the average false positive rate (FPR) is 4.47%, respectively. The preliminary results showed the feasibility and efficiency of the proposed method.

  18. Micro Flow Cytometer Chip Integrated with Micro-Pumps/Micro-Valves for Multi-Wavelength Cell Counting and Sorting

    NASA Astrophysics Data System (ADS)

    Chang, Chen-Min; Hsiung, Suz-Kai; Lee, Gwo-Bin

    2007-05-01

    Flow cytometry is a popular technique for counting and sorting of individual cells. This study presents a new chip-based flow cytometer capable of cell injection, counting and switching in an automatic format. The new microfluidic system is also capable of multi-wavelength detection of fluorescence-labeled cells by integrating multiple buried optical fibers within the chip. Instead of using large-scale syringe pumps, this study integrates micro-pumps and micro-valves to automate the entire cell injection and sorting process. By using pneumatic serpentine-shape (S-shape) micro-pumps to drive sample and sheath flows, the developed chip can generate hydrodynamic focusing to allow cells to pass detection regions in sequence. Two pairs of optical fibers are buried and aligned with the microchannels, which can transmit laser light sources with different wavelengths and can collect induced fluorescence signals. The cells labeled with different fluorescent dyes can be excited by the corresponding light source at different wavelengths. The fluorescence signals are then collected by avalanche photodiode (APD) sensors. Finally, a flow switching device composed of three pneumatic micro-valves is used for cell sorting function. Experimental data show that the developed flow cytometer can distinguish specific cells with different dye-labeling from mixed cell samples in one single process. The target cell samples can be also switched into appropriate outlet channels utilizing the proposed microvalve device. The developed microfluidic system is promising for miniature cell-based biomedical applications.

  19. Somatic cell counts in milk of Welsh-Mountain, Dorset-Horn and Chios ewes throughout lactation

    Microsoft Academic Search

    G. C. Fthenakis

    1996-01-01

    During four experiments the somatic cell counts (SCC) of 2642 milk samples were determined. SCC were studied in the milk of healthy Welsh-Mountain and Dorset-Horn ewes (experiment I), healthy Chios ewes (experiment II), Welsh-Mountain ewes subjected to intramammary inoculation with Staphylococcus simulans (experiment III) or Dorset-Horn ewes similarly inoculated (experiment IV). SCC of less than 1.0 × 106 cells ml?1

  20. Incidence of Clinical Mastitis in Dairy Herds Grouped in Three Categories by Bulk Milk Somatic Cell Counts

    Microsoft Academic Search

    H. W. Barkema; Y. H. Schukken; T. J. G. M. Lam; M. L. Beiboer; H. Wilmink; G. Benedictus; A. Brand

    1998-01-01

    Incidence of clinical mastitis was studied in 274 herds grouped in three categories by bulk milk so- matic cell count (SCC). Mean incidence rate of clini- cal mastitis was 0.278, 0.257, and 0.252 cases per 365 cow-days at risk in herds with low ( ?150,000), medium (150,000 to 250,000), and high (250,000 to 400,000 cells\\/ml) bulk milk SCC, respectively. The

  1. CD4 Cell Count Trends after Commencement of Antiretroviral Therapy among HIV-Infected Patients in Tigray, Northern Ethiopia: A Retrospective Cross-Sectional Study

    PubMed Central

    Asfaw, Addisu; Ali, Dagim; Eticha, Tadele; Alemayehu, Adissu; Alemayehu, Mussie; Kindeya, Filmon

    2015-01-01

    Background The rate and extent of CD4 cell recovery varies widely among HIV-infected patients with different baseline CD4 cell count strata. The objective of the study was to assess trends in CD4 cell counts in HIV-infected patients after initiation of antiretroviral therapy in Tigray, Northern Ethiopia. Methods A retrospective cross-sectional study was conducted by reviewing medical records of HIV patients who received antiretroviral treatment at twenty health centers in Tigray region during 2008–2012. Multi-stage cluster sampling technique was employed to collect data, and the data were analyzed using SPSS version 20.0 software. Results The median change from baseline to the most recent CD4 cell count was +292 cells/?l. By 5 years, the overall median (inter-quartile range, IQR) CD4 cell count was 444(263-557) cells/?l while the median (IQR) CD4 cell count was 342(246-580) cells/?l among patients with baseline CD4 cell counts ?200 cells/?l, 500(241-557) cells/?l among those with baseline CD4 cell counts of 201–350 cells/?l, and 652(537-767) cells/?l among those with baseline CD4 cell counts >350 cells/?l. Higher baseline CD4 cell counts and being male were independently associated with the risk of immunological non-response at 12 months. Furthermore, it was also investigated that these factors were significant predictors of subsequent CD4 cell recovery. Conclusions Patients with higher baseline CD4 cell stratum returned to normal CD4 Cell counts though they had an increased risk of immunological non-response at 12 months compared to those with the least baseline CD4 cell stratum. The findings suggest that consideration be given to initiation of HAART at a CD4 cell count >350 cells/?l to achieve better immune recovery, and to HIV-infected male patients to improve their health seeking behavior. PMID:25816222

  2. Multipotency of equine mesenchymal stem cells derived from synovial fluid.

    PubMed

    Murata, D; Miyakoshi, D; Hatazoe, T; Miura, N; Tokunaga, S; Fujiki, M; Nakayama, K; Misumi, K

    2014-10-01

    Cartilage regeneration with cell therapy following arthroscopic surgery could be used in racehorses with intra-articular fractures (IAF) and osteochondritis dissecans (OCD). The aims of this study were to investigate the origin and multipotency of stromal cells in the synovial fluid (SF) of horses with intra-articular injury and synovitis, and to provide a new strategy for regeneration of lost articular cartilage. Mesenchymal stromal cells were isolated from SF of horses with IAF and OCD. Multipotency was analysed by RT-PCR for specific mRNAs and staining for production of specific extracellular matrices after induction of differentiation. The total number of SF-derived mesenchymal stromal cells reached >1?×?10(7) by the fourth passage. SF-derived cells were strongly positive (>90% cells positive) for CD44, CD90 and major histocompatibility complex (MHC) class I, and moderately positive (60-80% cells positive) for CD11a/CD18, CD105 and MHC class II by flow cytometry. SF-derived cells were negative for CD34 and CD45. Under specific nutrient conditions, SF-derived cells differentiated into osteogenic, chondrogenic, adipogenic and tenogenic lineages, as indicated by the expression of specific marker genes and by the production of specific extracellular matrices. Chondrogenic induction in culture resulted in a change in cell shape to a 'stone-wall' appearance and formation of a gelatinous sheet that was intensely stained with Alcian blue. SF may be a novel source of multipotent mesenchymal stem cells with the ability to regenerate chondrocytes. PMID:25151209

  3. A cell-based sensor of fluid shear stress for microfluidics

    E-print Network

    Varma, Sarvesh

    2013-01-01

    Fluid flow is an essential feature of every microsystem involving cell handling, culture or sorting. The particular application determines the relevant flow rates used in a device. Flows inevitably generate fluid shear ...

  4. Factors affecting somatic cell counts and their relations with milk and milk constituent yield in buffaloes.

    PubMed

    Cerón-Muñoz, M; Tonhati, H; Duarte, J; Oliveira, J; Muñoz-Berrocal, M; Jurado-Gámez, H

    2002-11-01

    Data concerning daily milk yield (MY), percentage of milk fat (%F), protein (%P), lactose (%LT), and total solids (%TS), and somatic cell counts (SCC) for a herd of 222 Murrah buffalo reared in the state of São Paulo, Brazil, were collected monthly from 1997 to 2000 in order to study the factors affecting SCC and their relation to milk production and constituents during lactation. SCC decreased in the second month of lactation and increased thereafter, up to the ninth month of lactation. The interaction of month of lactation x order of calving was significant. Mean MY observed during the first month of lactation was 6.87 kg, which increased to 7.65 kg during the second month, and then decreased until the ninth month of lactation (3.83 kg). During the different months of lactation, %F, %P, %LT, and %TS ranged from 6.28 to 8.38%, 4.05 to 4.59%, 4.96 to 5.34%, and 16.94 to 18.55%, respectively. Calving year, calving order, and order of month of lactation significantly affected MY, %F, %P, %LT, and %TS. The regression coefficients of transformed SCC on MY and %LT were negative and significant during all months of lactation, showing that milk and lactose yield decreased with increased transformed SCC, causing losses to buffalo milk producers. PMID:12487456

  5. Serum Uric Acid, Alanine Aminotransferase, Hemoglobin and Red Blood Cell Count Levels in Pseudoexfoliation Syndrome

    PubMed Central

    Simavl?, Hüseyin; Bucak, Yasin Yücel; Tosun, Mehmet; Erdurmu?, Mesut

    2015-01-01

    Purpose. The pathogenesis of pseudoexfoliation (PEX), the most common cause of secondary glaucoma, has not been clearly identified, but there is increasing evidence that points out the role of oxidative stress. The aim of this study is to evaluate some of the most commonly used blood parameters, hemoglobin (Hb), red blood cell count (RBC), alanine aminotransferase (ALT), and uric acid (UA) levels, in subjects with PEX. Materials and Methods. This study is performed in a state hospital between November 2011 and December 2012. Retrospective chart review of subjects who underwent cataract surgery was performed. Thirty-one healthy subjects with PEX and 34 healthy subjects without PEX were evaluated. Hb, RBC, ALT, and UA levels were recorded. Student's t-test was used to compare the two groups. Results. The mean age was 73.6 ± 14.1 years in PEX group and 70.1 ± 12.7 in control group (p = 0.293). Hb, RBC, ALT, and UA levels did not show a statistically significant difference among PEX and control groups (p > 0.05 for all). Conclusion. Serum levels of Hb, RBC, ALT, and UA levels were similar in subjects with and without PEX. Further studies are needed to clarify the precise role of Hb, RBC, ALT, and UA in the pathogenesis of PEX.

  6. Presence of viral and bacterial organisms in milk and their association with somatic cell counts.

    PubMed

    Herlekar, D A; Shashikant, C S; Gurjar, A A; Jayarao, B M

    2013-10-01

    About 20 to 35% of milk samples from cows with intramammary infection or high somatic cell count (SCC) are negative on bacteriological culture analysis. However, little is known about SCC in milk of cows infected with viruses. In the first part of our study, we developed a real-time PCR assay for detection of bovine herpesvirus (BHV) 1, BHV2, and BHV4, and bovine viral diarrhea virus (BVDV) in composite quarter milk samples. A total of 1,479 lactating cows of 1,964 cows in the dairy herd were initially selected because these cows had complete SCC data for at least 3 consecutive test results, of which 139 lactating cows from different lactation age groups were selected randomly and studied extensively. Composite quarter milk samples were collected on 3 alternate days and examined for viruses, SCC, and bacteriological analysis. In total, 10, 28, and 0.7% of the composite quarter milk samples from cows were positive for BHV1, BHV2, and BHV4, respectively; BVDV was not detected in composite quarter milk samples. Bovine herpesvirus was not associated with a particular bacterial species. Our study results indicate that cows positive for BHV in composite quarter milk samples alone are less likely to have elevated SCC compared with cows with bacterial intramammary infection; BHV1, BHV2, and BHV4 are probably not major udder pathogens. PMID:23972495

  7. Coupling EELS/EFTEM Imaging with Environmental Fluid Cell Microscopy

    SciTech Connect

    Unocic, Raymond R [ORNL; Baggetto, Loic [ORNL; Veith, Gabriel M [ORNL; Dudney, Nancy J [ORNL; More, Karren Leslie [ORNL

    2012-01-01

    Insight into dynamically evolving electrochemical reactions and mechanisms encountered in electrical energy storage (EES) and conversion technologies (batteries, fuel cells, and supercapacitors), materials science (corrosion and oxidation), and materials synthesis (electrodeposition) remains limited due to the present lack of in situ high-resolution characterization methodologies. Electrochemical fluid cell microscopy is an emerging in-situ method that allows for the direct, real-time imaging of electrochemical processes within a fluid environment. This technique is facilitated by the use of MEMS-based biasing microchip platforms that serve the purpose of sealing the highly volatile electrolyte between two electron transparent SiNx membranes and interfacing electrodes to an external potentiostat for controlled nanoscale electrochemislly experiments [!]. In order to elucidate both stmctural and chemical changes during such in situ electrochemical experiments, it is impmtant to first improve upon the spatial resolution by utilizing energy-filtered transmission electron microscopy (EFTEM) (to minimize chromatic aben ation), then to detennine the chemical changes via electron energy loss spectroscopy (EELS). This presents a formidable challenge since the overall thickness through which electrons are scattered through the multiple layers of the cell can be on the order of hundreds of nanometers to microns, scattering through which has the deleterious effect of degrading image resolution and decreasing signal-to noise for spectroscopy [2].

  8. Effect of an automated dipping and backflushing system on somatic cell counts.

    PubMed

    Olde Riekerink, R G M; Ohnstad, I; van Santen, B; Barkema, H W

    2012-09-01

    Postmilking teat disinfection is an effective management practice to prevent transmission of contagious mastitis pathogens from cow to cow. With farms increasing in size and an increase in the number of rotary milking parlors, the need for automation of postmilking teat disinfection is mounting. Automated teat dipping and backflushing (ADB) systems have existed for some years, but their effect on udder health was never examined in a field study on commercial dairy farms. The objectives of this study were, therefore, to evaluate the effect of introducing an ADB system in a herd on (1) bulk milk somatic cell count (SCC), (2) individual cow SCC, and (3) the proportion of newly elevated SCC. Dairy herd improvement data were collected over a 30-mo period on 25 sets of 3 farms. Each set of 3 farms contained a farm that installed an ADB system, one that disinfected teats using dipping after milking, and one that sprayed teats after milking. Data were analyzed using linear mixed models. Bulk milk SCC on farms that sprayed or dipped before installing an ADB system were 16,000 and 30,000 cells/mL lower in the period 6 to 18 mo after installation, respectively, than on farms that continued spraying or dipping the teats after milking. In the same period after installing an ADB system, proportions of cows with elevated SCC were 4.3 and 1.2% lower, respectively, compared with spraying and with dipping. Similarly, proportions of cows that had newly elevated SCC were 1.5% lower and 0.3% higher, respectively, compared with farms that sprayed or dipped. Installing an ADB system had a beneficial effect on bulk milk SCC, individual cow SCC, and the proportion of newly elevated SCC. The effect was most prominent in the period 6 to 18 mo after installation of an ADB system. PMID:22916897

  9. Increased counts and degranulation of duodenal mast cells and eosinophils in functional dyspepsia- a clinical study.

    PubMed

    Song, Shijun; Song, Yan; Zhang, Haishan; Li, Gaiqin; Li, Xiaopei; Wang, Xiaohong; Liu, Zhen

    2015-02-01

    The above article published in Medicinski Glasnik online on 26 June 2014 by the Medical Association of Zenica-Doboj Canton (http://www.ljkzedo.com.ba/index.php/u-sljedecem-broju) and in Volume 11, Issue 2, pages 276-282, has been retracted by agreement between the authors, the journal Editor-in-Chief, Professor Selma Uzunovi?, and the Medical Association of Zenica-Doboj Canton. The reasons for this retraction are as follows: The work reported in the paper was about the role of duodenal eosinophils and mast cells in the pathogenesis of functional dyspepsia. Most of the experiments were carried out by a former member of the authors' team named Yuan Haipeng, who has left the team for more than two years. A high proportion of data in the paper had been reported in the doctoral dissertation of Yuan Haipeng in 2012, and the paper was published without the knowledge or permission of Yuan. Besides the data previously reported in the doctoral dissertation of Yuan Haipeng, the authors calculated the other data in the paper before the submission. However, it has come to the authors' attention that they had made quite a few mistakes due to a loss of the original data, which was not described in details in the dissertation. REFERENCE Shijun Song, Yan Song, Haishan Zhang, Gaiqin Li, Xiaopei Li, Xiaohong Wang, Zhen Liu. Increased counts and degranulation of duodenal mast cells and eosinophils in functional dyspepsia- a clinical study. Med Glas (Zenica) 2014; 11(2):276-82. PMID:25669347

  10. The relationship between dairy cow hygiene and somatic cell count in milk.

    PubMed

    Sant'anna, A C; Paranhos da Costa, M J R

    2011-08-01

    Corporal hygiene is an important indicator of welfare for dairy cows and is dependent on facilities, climate conditions, and the behavior of the animals. The objectives of this study were to describe how the hygiene conditions of dairy cows vary over time and to assess whether a relationship exists between hygiene and somatic cell count (SCC) in milk. Monthly hygiene evaluations were conducted on lactating cows in 2 dairy farms for 9 consecutive months, totaling 3,554 evaluations from 545 animals. Hygiene was measured using a 4-point scoring system (very clean, clean, dirty, and very dirty) for 4 areas of the animal's body (leg, flank, abdomen, and udder) and combining these scores to generate a composite cleanliness score. A total of 2,218 milk samples was analyzed from 404 cows to determine SCC and somatic cell linear scores (SCLS). Individual variation was observed in the hygiene of cows throughout the year, with the highest proportion of clean cows being observed in August and the lowest in January. In spite of this seasonal variation, approximately half (55.62%) of the cows displayed consistent cleanliness scores, with 45.86% of them remaining consistently clean (very clean or clean) and 9.76% remaining dirty (very dirty or dirty) over the course of the study. The very clean cows had the lowest SCLS, followed by the clean, dirty, and very dirty cows (no statistically significant differences were found between the latter 2 groups). The most critical months for cow hygiene were those with the greatest rainfall, when a reduction in the welfare of cows and higher SCC values were observed. The evaluation and control of dairy cow hygiene are useful in defining management strategies to reduce problems with milk and improve the welfare of the animals. PMID:21787920

  11. Effects of season and herd milk volume on somatic cell counts of Florida dairy farms.

    PubMed

    Ferreira, F C; De Vries, A

    2015-06-01

    Dairy farms in Florida produce less milk and milk with higher somatic cell counts (SCC) in the hot and humid summer. This has consequences for the interpretation of average milk quality. The objectives were to describe the associations of bulk tank SCC (BTSCC) with time of the year and the milk volume per farm. Monthly BTSCC and milk volume records from 84% (in 2012; n=1,308) and 77% (in 2013; n=1,200) of the 130 dairy farms in Florida were used. Data were analyzed separately per year. We calculated arithmetic averages of the BTSCC for each farm (ASCCf), each month (ASCCm), and each year (ASCCy). We used the milk volume to calculate a milk-weighted average for each farm (WSCCf), each month (WSCCm), and each year (WSCCy). Period 1 (P1) was defined as February, March, and April, and period 2 (P2) was defined as August, September, and October. These periods generally had the lowest and highest BTSCC throughout the year, respectively. Seasonality was expressed by the P2/P1 ratios of BTSCC and milk volume in both periods. In 2012 and 2013, 72 and 74% of the monthly milk volume observations were <400,000cells/mL. A clear seasonal pattern with lower milk volume and higher ASCCm during P2 was observed for most farms. The averages of the P2/P1 ratio of milk volume were 0.68 and 0.74 in 2012 and 2013. The averages of the P2/P1 ratio of SCC were 1.30 and 1.65 for 2012 and 2013, respectively. The WSCCy was 297,000cells/mL in 2012 and 274,000cells/mL in 2013. These values were 13 and 16% lower than the ASCCy in the respective years. In 2012, 82% of the farms shipped milk with a lower WSCCf than their ASCCf. In 2013, 97% of the farms shipped milk with a lower WSCCf than their ASCCf. The difference between a farm's WSCCf and its ASCCf tended to be greater in more-seasonal farms for BTSCC and milk volume. The WSCCm was lower than the ASCCm in every calendar month in both years. Collectively, these results show that the SCC of pooled milk from Florida was substantially lower than the arithmetic averages of monthly BTSCC values. Therefore, it should be made clear if the SCC is weighted by milk volume when "average" SCC results are reported. Programs to improve milk quality in Florida might be focused on conditions during August, September, and October because the BTSCC is then markedly increased on many farms. PMID:25795483

  12. Validation of a single-platform, volumetric, flow cytometry for CD4 T cell count monitoring in therapeutic mobile unit

    PubMed Central

    2012-01-01

    Background A mobile health unit may be useful to follow up adult and pediatric patients on antiretroviral treatment and living in remote areas devoid of laboratory facilities. The study evaluated the use of the simplified, robust, single-plateform, volumetric, pan-leucogating Auto40 flow cytometer (Apogee Flow Systems Ltd, Hemel Hempstead, UK) for CD4 T cell numeration in a mobile unit, compared against a reference flow cytometry method. Methods The therapeutic mobile unit of the Laboratoire National de Santé Hygiène Mobile, Yaoundé, Cameroon, was equipped with the Auto40. A FACSCalibur flow cytometer (Becton Dickinson Immuno-cytometry System, San Jose, CA, USA) was used as reference method. EDTA-blood samples from volunteers were first subjected to CD4 T cell count in the mobile unit, and an aliquot was sent within 4 hours to Centre International de Référence Chantal Biya, Yaoundé, for FACSCalibur assay. Results Two HIV screening campaigns with the mobile unit were organised in December 2009 and January 2010. The campaign in the suburb of Yaoundé which was 20 km from the reference laboratory included 188 volunteers comprising 93 children less than 5 years old. The campaign in Ambang Bikok (53 km far from Yaoundé) included 69 adult volunteers. In Yaoundé suburb, mean ± standard deviation (SD) CD4 T cell count was 996 ± 874 cells/?l by Auto40, and 989 ± 883 cells/?l by FACSCalibur; in Ambang Bikok, mean ± SD CD4 T cell count was 1041 ± 317 cells/?l by Auto40, and 1032 ± 294 cells/?l by FACSCalibur. Results by Auto40 and FACSCalibur were highly correlated in Yaoundé (r2 = 0.982) as in Ambang Bikok (r2 = 0.921). Bland-Altman analysis showed a close agreement between Auto40 and FACSCalibur results expressed in absolute count as in percentage in Yaoundé and Ambang Bikok. When pooling the 257 CD4 T cell count measurements, the Auto40 yielded a mean difference of +7.6 CD4 T cells/?l higher than by reference flow cytometry; and the sensitivity and specificity of Auto40 in enumerating absolute CD4 T cell counts of less than 200 cells/?l were 87% and 99%, respectively, and in enumerating absolute CD4 T cell counts of less than 350 cells/?l were 87% and 98%, respectively. The intrarun and interun precisions of the Auto40 assay assessed in the mobile unit were 5.5% and 7.9%, respectively. Conclusions The Auto40 flow cytometer installed in a therapeutic mobile unit and operated far from its reference laboratory gave a perfect correlation with the reference method, and could be useful in carrying out immunological monitoring of HIV-infected patients living in areas without access to laboratory facilities. PMID:22309994

  13. V?9V?2 T-Cell Polyfunctionality Is Differently Modulated in HAART-Treated HIV Patients according to CD4 T-Cell Count

    PubMed Central

    Casetti, Rita; De Simone, Gabriele; Sacchi, Alessandra; Rinaldi, Alessandra; Viola, Domenico; Agrati, Chiara; Bordoni, Veronica; Cimini, Eleonora; Tumino, Nicola; Besi, Francesca; Martini, Federico

    2015-01-01

    Alteration of ?? T-cell distribution and function in peripheral blood is among the earliest defects during HIV-infection. We asked whether the polyfunctional response could also be affected, and how this impairment could be associated to CD4 T-cell count. To this aim, we performed a cross-sectional study on HIV-infected individuals. In order to evaluate the polyfunctional-V?9V?2 T-cell response after phosphoantigen-stimulation, we assessed the cytokine/chemokine production and cytotoxicity by flow-cytometry in HAART-treated-HIV+ persons and healthy-donors. During HIV-infection V?9V?2-polyfunctional response quality is affected, since several V?9V?2 T-cell subsets resulted significantly lower in HIV+ patients in respect to healthy donors. Interestingly, we found a weak positive correlation between V?9V?2 T-cell-response and CD4 T-cell counts. By dividing the HIV+ patients according to CD4 T-cell count, we found that Low-CD4 patients expressed a lower number of two V?9V?2 T-cell subsets expressing MIP-1? in different combinations with other molecules (CD107a/IFN?) in respect to High-CD4 individuals. Our results show that the V?9V?2 T-cell-response quality in Low-CD4 patients is specifically affected, suggesting a direct link between innate V?9V?2 T-cells and CD4 T-cell count. These findings suggest that V?9V?2 T-cell quality may be indirectly influenced by HAART therapy and could be included in a new therapeutical strategy which would perform an important role in fighting HIV infection. PMID:26161861

  14. Absolute peripheral monocyte count at diagnosis predicts central nervous system relapse in diffuse large B-cell lymphoma

    PubMed Central

    Nitta, Hideaki; Terui, Yasuhito; Yokoyama, Masahiro; Mishima, Yuko; Nishimura, Noriko; Ueda, Kyoko; Kusano, Yoshiharu; Tsuyama, Naoko; Takeuchi, Kengo; Kanda, Yoshinobu; Hatake, Kiyohiko

    2015-01-01

    Recently, elevated peripheral blood monocyte counts at diagnosis have been shown to be an independent marker associated with poor prognosis in patients with both non-Hodgkin and Hodgkin lymphoma. In this study, we retrospectively analyzed the data from a total of 550 patients with diffuse large B-cell lymphoma and evaluated the relationship between central nervous system relapse and absolute monocyte counts at diagnosis. Twenty-six patients developed central nervous system relapse. The central nervous system relapse-free survival rate was significantly lower in patients with the absolute monocyte counts ?0.51×109/L (87.8% versus 96.4%; P<0.001). This association was independently significant after adjusting for other significant factors, including systemic relapse as a time-dependent covariate by multivariate analysis (hazard ratio 2.46; 95% confidence intervals 1.05–5.75; P=0.039). These results suggest that the absolute monocyte count at diagnosis is an independent significant risk factor for central nervous system relapse in patients with diffuse large B-cell lymphoma. PMID:25261092

  15. [Influence of fluid shear stress on cultured vascular endothelial cells].

    PubMed

    Takakuwa, O

    1990-03-01

    Vascular endothelial cells modulate their functions in response to hemodynamic forces such as fluid shear stress. In the present study, we applied shear to cultured bovine aortic endothelial cells (EC) by using a saecially designed apparatus and examined the effects of their homogenate and conditioned medium on such EC and smooth muscle cell (SMC) functions as adhesion, growth, migration or collagen synthesis. Cultured bovine aortic SMC were stimulated to adhere to wells and grow in the presence of EC conditioned medium. This conditioned medium had no effect on EC adhesion and growth. The activities of stimulating SMC adhesion and growth were almost the same in both EC conditioned medium obtained from static cultures and shear-loaded cultures. Studies with filters in a modified Boyden chamber showed that shear-loaded EC homogenate yielded stimulated SMC migration. Also shear-loaded EC cell layer contained increased amount of collagen compared with static EC cell layer. These observations indicate that:(a) EC secrets the substances which stimulate SMC adhesion and growth, but these functions are not affected by shear stress application, (b) EC produces SMC migration stimulators in response to shear stress, and (c) shear stress can enhance EC collagen synthesis. These results are relevant to EC response to hemodynamic forces and its role in the localization of atherosclerotic lesions in vivo. PMID:2365276

  16. Human amniotic fluid stem cell differentiation along smooth muscle lineage.

    PubMed

    Ghionzoli, Marco; Repele, Andrea; Sartiani, Laura; Costanzi, Giulia; Parenti, Astrid; Spinelli, Valentina; David, Anna L; Garriboli, Massimo; Totonelli, Giorgia; Tian, Jun; Andreadis, Stelios T; Cerbai, Elisabetta; Mugelli, Alessandro; Messineo, Antonio; Pierro, Agostino; Eaton, Simon; De Coppi, Paolo

    2013-12-01

    Functional smooth muscle engineering requires isolation and expansion of smooth muscle cells (SMCs), and this process is particularly challenging for visceral smooth muscle tissue where progenitor cells have not been clearly identified. Herein we showed for the first time that efficient SMCs can be obtained from human amniotic fluid stem cells (hAFSCs). Clonal lines were generated from c-kit(+) hAFSCs. Differentiation toward SM lineage (SMhAFSCs) was obtained using a medium conditioned by PDGF-BB and TGF-?1. Molecular assays revealed higher level of ? smooth muscle actin (?-SMA), desmin, calponin, and smoothelin in SMhAFSCs when compared to hAFSCs. Ultrastructural analysis demonstrated that SMhAFSCs also presented in the cytoplasm increased intermediate filaments, dense bodies, and glycogen deposits like SMCs. SMhAFSC metabolism evaluated via mass spectrometry showed higher glucose oxidation and an enhanced response to mitogenic stimuli in comparison to hAFSCs. Patch clamp of transduced hAFSCs with lentiviral vectors encoding ZsGreen under the control of the ?-SMA promoter was performed demonstrating that SMhAFSCs retained a smooth muscle cell-like electrophysiological fingerprint. Eventually SMhAFSCs contractility was evident both at single cell level and on a collagen gel. In conclusion, we showed here that hAFSCs under selective culture conditions are able to give rise to functional SMCs. PMID:23995291

  17. Effect of sensor systems for cow management on milk production, somatic cell count, and reproduction.

    PubMed

    Steeneveld, W; Vernooij, J C M; Hogeveen, H

    2015-06-01

    To improve management on dairy herds, sensor systems have been developed that can measure physiological, behavioral, and production indicators on individual cows. It is not known whether using sensor systems also improves measures of health and production in dairy herds. The objective of this study was to investigate the effect of using sensor systems on measures of health and production in dairy herds. Data of 414 Dutch dairy farms with (n=152) and without (n=262) sensor systems were available. For these herds, information on milk production per cow, days to first service, first calving age, and somatic cell count (SCC) was provided for the years 2003 to 2013. Moreover, year of investment in sensor systems was available. For every farm year, we determined whether that year was before or after the year of investment in sensor systems on farms with an automatic milking system (AMS) or a conventional milking system (CMS), or whether it was a year on a farm that never invested in sensor systems. Separate statistical analyses were performed to determine the effect of sensor systems for mastitis detection (color, SCC, electrical conductivity, and lactate dehydrogenase sensors), estrus detection for dairy cows, estrus detection for young stock, and other sensor systems (weighing platform, rumination time sensor, fat and protein sensor, temperature sensor, milk temperature sensor, urea sensor, ?-hydroxybutyrate sensor, and other sensor systems). The AMS farms had a higher average SCC (by 12,000cells/mL) after sensor investment, and CMS farms with a mastitis detection system had a lower average SCC (by 10,000cells/mL) in the years after sensor investment. Having sensor systems was associated with a higher average production per cow on AMS farms, and with a lower average production per cow on CMS farms in the years after investment. The most likely reason for this lower milk production after investment was that on 96% of CMS farms, the sensor system investment occurred together with another major change at the farm, such as a new barn or a new milking system. Most likely, these other changes had led to a decrease in milk production that could not be compensated for by the use of sensor systems. Having estrus detection sensor systems did not improve reproduction performance. Labor reduction was an important reason for investing in sensor systems. Therefore, economic benefits from investments in sensor systems can be expected more from the reduction in labor costs than from improvements in measures of health and production in dairy herds. PMID:25841965

  18. Prognostic effect of peripheral blood cell counts in advanced diffuse large B-cell lymphoma treated with R-CHOP-like chemotherapy: A single institution analysis

    PubMed Central

    YAMAUCHI, TAKAHIRO; TASAKI, TOSHIKI; TAI, KATSUNORI; IKEGAYA, SATOSHI; TAKAGI, KAZUTAKA; NEGORO, EIJU; KISHI, SHINJI; YOSHIDA, AKIRA; IWASAKI, HIROMICHI; UEDA, TAKANORI

    2015-01-01

    The primary objective of the present study was to correlate blood cell counts (lymphocyte, monocyte and platelet counts) with early disease relapse following the attainment of complete remission (CR) by the rituximab, cyclophosphamide, doxorubicin, vincristine and prednisolone (R-CHOP)-like regimen in patients with advanced diffuse large B-cell lymphoma (DLBCL). In total, 30 patients were evaluated, with a median follow-up period of 43 months. All the participating patients attained CR. In total, eight patients experienced relapse within two years of the diagnosis, and the three-year overall survival rate was recorded as 77%. The peripheral counts for lymphocytes, monocytes and platelets, and the lymphocyte-monocyte ratio, all of which have been reported to be prognostic in DLBCL, were assessed. None of these parameters were correlated with the incidence of early relapse or with the prognosis. The lymphocyte count was higher in the patients with durable remission than in those who relapsed, however, no significant differences were identified. Thus, the present study concluded that early disease relapse was not predicted by peripheral blood cell counts in advanced DLBCL that reached CR using the R-CHOP-like regimen. PMID:25621059

  19. Cluster of differentiation 4+ cell count mean value, reference range and its influencing factors in Human Immunodeficiency Virus-seronegative pregnant women in Lagos

    PubMed Central

    Akinbami, A. A.; Dosunmu, A. O.; Adediran, A.; Adewunmi, A. A.; Rabiu, K. A.; Osunkalu, V.; Ajibola, S.; Uche, E. I.; Adelekan, A.

    2014-01-01

    Background: Immunity in pregnancy is physiologically compromised and this may affect cluster of differentiation four (CD4) count levels. It is well established that several factors affect CD4 count level in pregnancy. This study aims to determine the effects of maternal age, gestational age, parity and level of education as they influence CD4 count in pregnancy and also to determine the mean and reference range of CD4 count in pregnancy in Lagos, Nigeria. Materials and Methods: A descriptive cross-sectional study was carried out at Ante-natal clinics in Lagos State, Nigeria. About 5 mls of blood was collected into Ethylene Diamine Tetracetic Acid (EDTA) bottles from HIV-negative pregnant women in various gestational ages of pregnancy. CD4+ cell count and full blood count of all samples were done within 3 hours of collection. The descriptive data was given as means ± standard deviation (SD). Pearson's chi-squared test and correlation were used for analytical assessment. Results: A total of 74 pregnant women were recruited. The age range was 19–41 years and a mean age of 30.42 ± 5.34 years. The CD4+ cell count was not statistically significant when compared with participants ages P = 0.417, neither with gestational ages P = 0.323, nor with parity P = 0.247 nor level of education P = 0.96. An overall mean CD4+ cell count was 771.96 ± 250 cells/?l and the range was 193–1370 cells/?l. Conclusion: Maternal age, gestational age, parity and level of education had no significant effects on CD4+ cell count levels in pregnancy. The mean CD4+ cell count of HIV-negative pregnant women in Lagos is 771.96 ± 250 cells/?l. PMID:24791043

  20. ELISPOTs Produced by CD8 and CD4 Cells Follow Log Normal Size Distribution Permitting Objective Counting

    PubMed Central

    Karulin, Alexey Y.; Karacsony, Kinga; Zhang, Wenji; Targoni, Oleg S.; Moldovan, Ioana; Dittrich, Marcus; Sundararaman, Srividya; Lehmann, Paul V.

    2015-01-01

    Each positive well in ELISPOT assays contains spots of variable sizes that can range from tens of micrometers up to a millimeter in diameter. Therefore, when it comes to counting these spots the decision on setting the lower and the upper spot size thresholds to discriminate between non-specific background noise, spots produced by individual T cells, and spots formed by T cell clusters is critical. If the spot sizes follow a known statistical distribution, precise predictions on minimal and maximal spot sizes, belonging to a given T cell population, can be made. We studied the size distributional properties of IFN-?, IL-2, IL-4, IL-5 and IL-17 spots elicited in ELISPOT assays with PBMC from 172 healthy donors, upon stimulation with 32 individual viral peptides representing defined HLA Class I-restricted epitopes for CD8 cells, and with protein antigens of CMV and EBV activating CD4 cells. A total of 334 CD8 and 80 CD4 positive T cell responses were analyzed. In 99.7% of the test cases, spot size distributions followed Log Normal function. These data formally demonstrate that it is possible to establish objective, statistically validated parameters for counting T cell ELISPOTs. PMID:25612115

  1. The cleavage pattern of the axolotl egg studied by cinematography and cell counting

    Microsoft Academic Search

    K. Hara

    1977-01-01

    The temporal pattern of cleavage in the egg of the axolotl,Ambystoma mexicanum, was studied 1. by time-lapse microcinematography, and 2. by counting the total number of blastomeres dissociated at successive stages.

  2. Letters to Analytical Chemistry Microfluidic CD4+ T-Cell Counting Device Using

    E-print Network

    Sia, Samuel K.

    This letter demonstrates a microfluidic platform for enumerating CD4+ T-lymphocytes from whole blood using are based on clinical symptoms and absolute CD4 counts.7 In addition to identifying the onset of AIDS in HIV

  3. [Reduced performance and high somatic cell counts in a dairy herd fed high amounts of brewers' grain].

    PubMed

    Wenzinger, B

    2013-09-01

    The present case report describes a herd problem on a Holstein Friesian dairy farm in Switzerland, which could be attributed to the feeding of high amounts of wet brewers' grain over several months. Apathy and reduced general appearance, reduced feed intake as well as a decline in milk yield could be observed. A strong increase in milk somatic cell counts as well as an increase in the incidence of mastitis could be found. The milk fat content was highly elevated in all cows, whereas the milk protein content was reduced. The exclusion of wet brewers' grain from the partial mixed ration resulted in a considerable improvement of the general appearance of the cows and a decrease of the milk somatic cell counts. Feed that is easily spoiled could be a health risk for animals, particularly under hot and humid weather conditions and if fed in high amounts. PMID:23985095

  4. Determination of the abundance of cosmic matter via the cell count moments of the galaxy distribution

    NASA Astrophysics Data System (ADS)

    Bel, J.; Marinoni, C.

    2014-03-01

    We demonstrate that accurate and precise information about the matter content of the universe can be retrieved via a simple cell count analysis of the 3D spatial distribution of galaxies. A new clustering statistic, the galaxy clustering ratio?, is the key to this process. This is defined as the ratio between one- and two-point second-order moments of the smoothed galaxy density distribution. The distinguishing feature of this statistic is its universality: on large cosmic scales both galaxies (in redshift space) and mass (in real space) display the same ? amplitude. This quantity, in addition, does not evolve as a function of redshift. As a consequence, the ? statistic provides insight into characteristic parameters of the real-space power spectrum of mass density fluctuations without the need to specify the galaxy biasing function, neither a model for galaxy redshift distortions, nor the growing mode of density ripples. We demonstrate the method with the luminous red galaxy (LRG) sample extracted from the spectroscopic Sloan Digital Sky Survey (SDSS) data release 7 (DR7) catalogue. Taking weak (flat) priors of the curvature of the universe (?k) and of the constant value of the dark energy equation of state (w), and strong (Gaussian) priors of the physical baryon density ?bh2, of the Hubble constant H0, and of the spectral index of primordial density perturbations ns, we estimate the abundance of matter with a relative error of 8% (?m=0.283±0.023). We expect that this approach will be instrumental in searching for evidence of new physics beyond the standard model of cosmology and in planning future redshift surveys, such as BigBOSS or EUCLID.

  5. COMPARISON OF SOMATIC CELL COUNTS, MILK CONSTITUENTS AND WEANING WEIGHTS IN EWES WITH AND WITHOUT SUBCLINICAL MASTITIS

    Microsoft Academic Search

    A. B. Ciliax; D. W. Holcombe; D. Redelman; G. C. J. Fernandez

    Fifty-four multiparous Rambouillet ewes and their offspring were used to 1) assess the effectiveness of using flow cytometry (FC) relative to traditional somatic cell counts (SCC) and to determine the incidence of subclinical mastitis at weaning and 2) assess the relationships among FC, milk constituents, weaning weights and subclinical mastitis. At weaning, lambs were weighed and 15-mL milk samples were

  6. The contribution of mammary infections by coagulase-negative staphylococci to the herd bulk milk somatic cell count

    Microsoft Academic Search

    P. Rainard; M. Ducelliez; B. Poutrel

    1990-01-01

    Quarter foremilk samples were taken at 2–3 weekly intervals for several years in a experimental herd comprising about 45 cows. The samples were submitted to bacteriological analysis and somatic cell counting. The most prevalent quarter infections from 1982 to 1988 were by coagulase-negative staphylococci (15–20% of all the quarters sampled). Most of these (75.6%) persisted until drying-off Dry cow therapy

  7. The influence of somatic cell count in milk on reproductive traits and production of Black-and-White cows

    Microsoft Academic Search

    Vida Juozaitiene; Arunas Juozaitis

    2005-01-01

    The aim of this study was to estimate the relationships between somatic cell count in milk, reproduction traits and milk production of cows of the Black-and-White breed. The research was carried out on 412 herds of the active Black-and-White cattle population in Lithuania. Data on cows in the first three lactations during the period between 1998 and 2003 were used.

  8. Marginal Structural Models for Estimating the Effect of Highly Active Antiretroviral Therapy Initiation on CD4 Cell Count

    Microsoft Academic Search

    Stephen R. Cole; Miguel A. Hernan; Joseph B. Margolick; Mardge H. Cohen

    2005-01-01

    The effect of highly active antiretroviral therapy (HAART) on the evolution of CD4-positive T-lymphocyte (CD4 cell) count among human immunodeficiency virus (HIV)-positive participants was estimated using inverse probability-of-treatment-and-censoring (IPTC)-weighted estimation of a marginal structural model. Of 1,763 eligible participants from two US cohort studies followed between 1996 and 2002, 60 percent initiated HAART. The IPTC- weighted estimate of the difference

  9. CD4 T Cell Survival Following Intermittent IL-2 Therapy is Predictive of Increase in CD4 T Cell Count in HIV-Infected Patients

    PubMed Central

    Read, Sarah W.; Lempicki, Richard A.; Di Mascio, Michele; Srinivasula, Sharat; Burke, Rosanne; Sachau, William; Bosche, Marjorie; Adelsberger, Joseph W.; Sereti, Irini; Davey, Richard T.; Tavel, Jorge A.; Huang, Chiung-Yu; Issaq, Haleem J.; Fox, Stephen D.; Clifford Lane, H.; Kovacs, Joseph A.

    2009-01-01

    Administration of IL-2 to HIV-infected patients leads to significant increases in CD4 T cell counts. Previously we have shown that IL-2 induces increased proliferation and survival of CD4 T cells. Deuterium labeling studies were undertaken to study the relationship between IL-2 induced increases in CD4 T cell numbers and IL-2 effects on cell proliferation and survival. A strong inverse correlation was seen between the decay rate of label in CD4 cells and increases in CD4 cell numbers (R= -0.67; p<0.001). This correlation was not seen with the level of proliferating cells. Although the baseline CD4 cell count and number of CD4 cells expressing CD25 were also predictive of CD4 cell increases, the decay rate remained the most statistically significant predictor in multivariate regression models. Thus, increase in survival of CD4 T cells appears to be the critical mechanism leading to sustained CD4 cell increases in HIV-infected patients receiving intermittent IL-2 therapy. PMID:18684102

  10. Evaluation of the overall accuracy of the DeLaval cell counter for somatic cell counts in ovine milk.

    PubMed

    Gonzalo, C; Linage, B; Carriedo, J A; de la Fuente, F; Primitivo, F San

    2006-12-01

    The DeLaval cell counter (DCC) is a portable device designed for on-farm somatic cell count (SCC) analysis in bovine milk. This study evaluated the performance of the DCC when analyzing ovine milk. A total of 29 composite ovine milk samples, ranging between 20 x 10(3) and 2,200 x 10(3) cells/mL, were divided into 15 aliquots/milk sample corresponding to 5 SCC methods using 3 types of preservation (unpreserved, azidiol, and bronopol). The SCC methods were the Fossomatic (FSCC), the DCC in undiluted samples, and the DCC in samples diluted 1:1 in 3 different types of diluents (PBS + Triton X-100, PBS + ethidium bromide + Triton X-100, and PBS + propidium iodide + Triton X-100). All analyses were carried out in duplicate. In addition, each sample was analyzed in quadruplicate by the direct microscopic method (DMSCC) using Pyronin Y-methyl green as a stain. Comparison of methods was based on overall accuracy studies (means comparison, repeatability, and regression studies vs. DMSCC and FSCC as reference methods). The DCC methods used to analyze milk samples diluted in staining solution (with ethidium bromide or propidium iodide) showed large coefficients of regression (b = 0.91 to 1.01) and correlation (r > 0.99) when compared with the DMSCC and FSCC methods. In these samples the DCC gave repeatability values (s(r) = 33 to 48 x 10(3) cells/mL) similar to the DMSCC (s(r) = 36 x 10(3) cells/mL), and their log SCC means (5.52 to 5.54) did not differ from the reference value (5.54). However, undiluted samples analyzed by the DCC method showed large standard deviations of repeatability and SCC values lower than those by the DMSCC or FSCC methods, probably because of the high solids content in ovine milk. The type of preservation did not affect the outcomes. Consequently, the DCC was determined to be accurate when analyzing diluted ovine milk based on comparison with the SCC reference methods. PMID:17106093

  11. Reduction in Preterm Delivery and Neonatal Mortality after the Introduction of Antenatal Cotrimoxazole Prophylaxis among HIV-Infected Women with Low CD4 Cell Counts

    PubMed Central

    Walter, Jan; Mwiya, Mwiya; Scott, Nancy; Kasonde, Prisca; Sinkala, Moses; Kankasa, Chipepo; Kauchali, Shuaib; Aldrovandi, Grace M.; Thea, Donald M.; Kuhn, Louise

    2006-01-01

    Background. Cotrimoxazole prophylaxis is recommended for subgroups of human immunodeficiency virus (HIV)-infected adults and children to reduce all-cause morbidity and mortality. We investigated whether antenatal cotrimoxazole prophylaxis begun during pregnancy for HIV-infected pregnant women with low CD4 cell counts would affect birth outcomes. Methods. Cotrimoxazole prophylaxis was introduced as a routine component of antenatal care for HIV-infected women with CD4 cell counts <200 cells/?L during the course of a trial of mother-to-child HIV transmission in Lusaka, Zambia. Rates of preterm delivery, low birth weight, and neonatal mortality were compared for women with low CD4 cell counts before and after its introduction. Results. Among 255 women with CD4 cell counts <200 cells/?L, the percentage of preterm births (?34 weeks of gestation) was lower (odds ratio [OR], 0.49 [95% confidence interval {CI}, 0.24-0.98]) after cotrimoxazole prophylaxis was introduced than before; there was a significant decrease in neonatal mortality (9% to 0%; P = .01) and a trend toward increased birth weight (? = 114 g [95% CI, -42 to 271 g]). In contrast, there were no significant changes in these parameters over the same time interval among women with CD4 cell counts ?200 cells/?L. Conclusion. Antenatal provision of cotrimoxazole for HIV-infected pregnant women with low CD4 cell counts may have indirect benefits for neonatal health. PMID:17083035

  12. Demonstration of mast cell chemotactic activity in synovial fluid from rheumatoid patients

    PubMed Central

    Olsson, N; Ulfgren, A; Nilsson, G

    2001-01-01

    OBJECTIVES—The significance of the mast cell in the pathogenesis of rheumatic diseases has become more evident. Although mast cell hyperplasia is a feature of rheumatoid arthritis, the nature of mast cell chemoattractants involved in the recruitment of mast cells in joint diseases has not been studied in any detail. In this study the presence of mast cell chemotactic activity in synovial fluids was examined.?METHODS—Synovial fluids from seven rheumatoid patients were tested in a modified Boyden chamber, where a human mast cell line was used as responder. The presence of stem cell factor (SCF) and transforming growth factor ? (TGF?) was measured by enzyme linked immunosorbent assay (ELISA).?RESULTS—Six of the seven synovial fluids tested exhibited mast cell chemotactic activity. Two characterised human mast cell chemotaxins, SCF and TGF?, were highly expressed in the synovium. Soluble SCF could be detected in all fluids analysed. Blocking antibodies against SCF or TGF? almost completely blocked the activity in one fluid, partially blocked the activity in three, and did not affect the activity in two. Treatment of the responder cells with pertussis toxin reduced the migratory response against seven fluids, indicating the presence of chemoattractants mediating their effect through Gi coupled receptors.?CONCLUSION—These data demonstrate the presence of multiple factors in synovial fluid acting as mast cell chemoattractants, two of which are SCF and TGF? that contribute to the effect. These findings may be of importance for developing new strategies to inhibit mast cell accumulation in rheumatic diseases.?? PMID:11171676

  13. White Blood Cell Counts in Persons Aged 65 Years or More from the Cardiovascular Health Study Correlations with Baseline Clinical and Demographic Characteristics

    Microsoft Academic Search

    Edwin G. Bovill; Diane E. Bild; Gerardo Heiss; Lewis H. Kuller; Marshall H. Lee; Robert Rock; Patricia W. Wahl

    A higher white blood cell (WBC) count has been shown to be a risk factor for myocardial infarction and stroke in middle-aged populations. This study evaluated the relation between baseline WBC count and other risk factors, as well as subclinical and prevalent disease, in the Cardiovascular Health Study, an epidemiologic study of coronary heart disease and stroke in 5,201 persons

  14. Baseline total and specific differential white blood cell counts and 5-year all-cause mortality in community-dwelling older women

    Microsoft Academic Search

    Sean X. Leng; Qian-Li Xuea; Yi Huang; Luigi Ferrucci; Linda P. Fried; Jeremy D. Walston

    Increasing evidence demonstrates that inflammation is associated with many pathophysiologic processes and mortality in older adults. Increase in total white blood cell (WBC) counts is recognized as an important cellular marker of systemic inflammation. However, relationships of total WBC and individual differential counts with mortality in older adults, particularly in older women, have not been adequately evaluated. To address this

  15. Modeling centrifugal cell washers using computational fluid dynamics.

    PubMed

    Kellet, Beth E; Han, Binbing; Dandy, David S; Wickramasinghe, S Ranil

    2004-11-01

    Reinfusion of shed blood during surgery could avoid the need for blood transfusions. Prior to reinfusion of the red blood cells, the shed blood must be washed in order to remove leukocytes, platelets, and other contaminants. Further, the hematocrit of the washed blood must be increased. The feasibility of using computational fluid dynamics (CFD) to guide the design of better centrifuges for processing shed blood is explored here. The velocity field within a centrifuge bowl and the rate of protein removal from the shed blood has been studied. The results obtained indicate that CFD could help screen preliminary centrifuge bowl designs, thus reducing the number of initial experimental tests required when developing new centrifuge bowls. Although the focus of this work is on washing shed blood, the methods developed here are applicable to the design of centrifuge bowls for other blood-processing applications. PMID:15504118

  16. Associations of dairy cow behavior, barn hygiene, cow hygiene, and risk of elevated somatic cell count.

    PubMed

    Devries, T J; Aarnoudse, M G; Barkema, H W; Leslie, K E; von Keyserlingk, M A G

    2012-10-01

    Poor dairy cow hygiene has been consistently associated with elevated somatic cell count (SCC) and the risk of subclinical mastitis. The objective of this study was to determine the associations between dairy cow standing and lying behavior, barn hygiene, cow hygiene, and the risk of experiencing elevated SCC. Lactating Holstein dairy cows (n=69; 86 ± 51 DIM; parity: 2.0 ± 1.2; means ± SD), kept in 1 of 2 groups, were monitored over a 4-mo period. Each group contained 61 ± 1 (mean ± SD) cows over the study period; complete data were obtained from 37 and 32 animals within each respective group. Cows were housed in a sand-bedded, freestall barn with 2 symmetrical pens, each with a free cow traffic automatic milking system. To vary barn hygiene, in 4 consecutive 28-d periods, alley manure scrapers in each of the 2 pens were randomly assigned to frequencies of operation of 3, 6, 12, and 24 times per day. During the last 7 d of each period, cow hygiene (upper leg/flank, lower legs, and udder; scale of 1 = very clean to 4 = very dirty) and stall hygiene (number of 0.15×0.15-m squares contaminated with manure in a 1.20×1.65-m grid) were recorded. Standing and lying behavior of the cows were collected during those days using data loggers. Individual-cow SCC was recorded at the beginning and end of each 28-d period. Elevated SCC was used as an indicator of subclinical mastitis; incidence of elevated SCC was defined as having a SCC >200,000 cells/mL at the end of each 28-d period, when SCC was <100,000 cells/mL at the beginning of the period. Less frequent scraping of the barn alleys was associated with cows having poorer hygiene. Poor udder hygiene was associated with poor stall hygiene. Longer lying duration was associated with poor hygiene of the upper legs/flank and udder. Greater premilking standing duration was associated with poor udder hygiene and decreased frequency of lying bouts was associated with poor hygiene of the lower legs. Higher milk yield was associated with poor hygiene of the udder and lower legs; multiparous cows had poorer hygiene of the upper legs/flank and udder. Over the study period, 24 new cases of elevated SCC were detected. No associations existed for the risk of experiencing an elevated SCC with alley scraping frequency or cow behavior patterns. However, increased odds of occurrence of elevated SCC were noted for cows of lower milk yield as well as for multiparous cows. In summary, these results show that cow hygiene is affected by the standing and lying behavior of cows and by the cleanliness of the cow's environment. These findings emphasize the need for cows to be provided clean standing and lying environments. The results also show that frequent cleaning of barn alley floors will help improve cow hygiene. PMID:22884345

  17. An improved micro-extraction cell for supercritical fluid extraction and chromatography of fatty acids

    Microsoft Academic Search

    William E. Artz; Robert M. Sauer

    1992-01-01

    An improved supercritical fluid micro-extraction cell of increased reliability was designed for on-line supercritical fluid\\u000a extraction and chromatography (SFE\\/SFC) of food and other lipid-related samples. The key components in the modified cell include\\u000a a Swagelok stainless steel reducing union with a dual ferrule as the cell, with polyetherether-ketone (PEEK) ferrules and\\u000a nuts to connect the cell to the control valve.

  18. Highly sensitive automated method for DNA damage assessment: gamma-H2AX foci counting and cell cycle sorting.

    PubMed

    Hernández, Laia; Terradas, Mariona; Martín, Marta; Tusell, Laura; Genescà, Anna

    2013-01-01

    Phosphorylation of the H2AX protein is an early step in the double strand break (DSB) repair pathway; therefore, phosphorylated histone (?H2AX) foci scoring is widely used as a measure for DSBs. Foci scoring is performed either manually or semi-automatically using hand-operated capturing and image analysis software. In general, both techniques are laborious and prone to artifacts associated with manual scoring. While a few fully automated methods have been described in the literature, none of them have been used to quantify ?H2AX foci in combination with a cell cycle phase analysis. Adding this feature to a rapid automated ?H2AX foci quantification method would reduce the scoring uncertainty that arises from the variations in the background level of the ?H2AX signal throughout the cell cycle. The method was set up to measure DNA damage induced in human mammary epithelial cells by irradiation under a mammogram device. We adapted a FISH (fluorescent in situ hybridization) Spot-counting system, which has a slide loader with automatic scanning and cell capture system throughout the thickness of each cell (z-stack), to meet our assay requirements. While scanning the sample, the system classifies the selected nuclei according to the signal patterns previously described by the user. For our purposes, a double staining immunofluorescence was carried out with antibodies to detect ?H2AX and pericentrin, an integral component of the centrosome. We could thus distinguish both the number of ?H2AX foci per cell and the cell cycle phase. Furthermore, restrictive settings of the program classifier reduced the "touching nuclei" problem described in other image analysis software. The automated scoring was faster than and as sensitive as its manually performed counterpart. This system is a reliable tool for ?H2AX radio-induced foci counting and provides essential information about the cell cycle stage. It thus offers a more complete and rapid assessment of DNA damage. PMID:23903043

  19. Has the Rate of CD4 Cell Count Decline before Initiation of Antiretroviral Therapy Changed over the Course of the Dutch HIV Epidemic among MSM?

    PubMed Central

    Gras, Luuk; Geskus, Ronald B.; Jurriaans, Suzanne; Bakker, Margreet; van Sighem, Ard; Bezemer, Daniela; Fraser, Christophe; Prins, Jan M.; Berkhout, Ben

    2013-01-01

    Introduction Studies suggest that the HIV-1 epidemic in the Netherlands may have become more virulent, leading to faster disease progression if untreated. Analysis of CD4 cell count decline before antiretroviral therapy (ART) initiation, a surrogate marker for disease progression, may be hampered by informative censoring as ART initiation is more likely with a steeper CD4 cell count decline. Methods Development of CD4 cell count from 9 to 48 months after seroconversion was analyzed using a mixed-effects model and 2 models that jointly modeled CD4 cell counts and time to censoring event (start ART, <100 CD4 cells/mm3, or AIDS) among therapy-naïve MSM HIV-1 seroconverters in the Netherlands. These models make different assumptions about the censoring process. Results All 3 models estimated lower median CD4 cell counts 9 months after seroconversion in later calendar years (623, 582, and 541 cells/mm3 for 1984–1995 [n?=?111], 1996–2002 [n?=?139], and 2003–2007 seroconverters [n?=?356], respectively, shared-parameter model). Only the 2 joint-models found a trend for a steeper decline of CD4 cell counts with seroconversion in later calendar years (overall p-values 0.002 and 0.06 for the pattern-mixture and the shared-parameter model, respectively). In the shared-parameter model the median decline from 9 to 48 months was 276 cellsmm3 for 1984–1995 seroconverters and 308 cells/mm3 for 2003–2007 seroconverters (difference in slope, p?=?0.045). Conclusion Mixed-effects models underestimate the CD4 cell decline prior to starting ART. Joint-models suggest that CD4 cell count declines more rapidly in patients infected between 2003 and 2007 compared to patients infected before 1996. PMID:23724048

  20. Association of White Blood Cell Count and C-Reactive Protein with Outcomes in Children Hospitalized for Community-acquired Pneumonia.

    PubMed

    Williams, Derek J; Hall, Matthew; Auger, Katherine A; Tieder, Joel S; Jerardi, Karen E; Queen, Mary Ann; Statile, Angela M; Myers, Angela L; Shah, Samir S

    2015-07-01

    We examined the association between baseline peripheral white blood cell count and C-reactive protein (CRP) values with outcomes among 153 children hospitalized with pneumonia. In multivariable analyses, CRP, but not white blood cell count, was significantly associated with both fever duration and hospital length of stay. For every 1mg/dL increase in CRP, length of stay increased by 1 hour. PMID:25961893

  1. Relationship between antiretrovirals used as part of a cART regimen and CD4 cell count increases in patients with suppressed viremia

    Microsoft Academic Search

    Amanda Mocroft; Andrew N Phillips; Bruno Ledergerber; Christine Katlama; Antonio Chiesi; Frank-Detlef Goebel; Brygioa Knysz; Francisco Antunes; Peter Reiss; Jens D Lundgren

    2006-01-01

    Background: It is unknown if the CD4 cell count response differs according to antiretroviral drugs used in combination antiretroviral therapy (cART) in patients with maximal virological suppression [viral load (VL) < 50 copies\\/ml]. Objectives: To compare the change in CD4 cell count over consecutive measurements with VL < 50 copies\\/ml at both time-points according to nucleoside backbones and other antiretrovirals

  2. T E C H N I C A L R E P O R T Single-molecule transcript counting of stem-cell

    E-print Network

    van Oudenaarden, Alexander

    T E C H N I C A L R E P O R T Single-molecule transcript counting of stem-cell markers in the mouse the molecular identities of adult stem cells requires technologies for sensitive transcript detection in tissues distinct stem-cell populations, residing either at crypt bases or at position +4, but a detailed analysis

  3. Quantitative Assessment of Intra-Patient Variation in CD4+ T Cell Counts in Stable, Virologically-Suppressed, HIV-Infected Subjects

    PubMed Central

    Gordon, Claire L.; Cheng, Allen C.; Cameron, Paul U.; Bailey, Michael; Crowe, Suzanne M.; Mills, John

    2015-01-01

    Objectives Counts of absolute CD4+ T lymphocytes (CD4+ T cells) are known to be highly variable in untreated HIV-infected individuals, but there are no data in virologically-suppressed individuals. We investigated CD4+ T cell variability in stable, virologically-suppressed, HIV-1 infected adults on combination antiretroviral therapy (cART). Methods From a large hospital database we selected patients with stable virological suppression on cART for >3 years with >10 CD4+ T cell measurements performed over a further >2 years; and a control group of 95 patients not on cART. Results We identified 161 HIV-infected patients on cART without active HCV or HBV infection, with stable virological suppression for a median of 6.4 years. Over the study period 88 patients had reached a plateau in their absolute CD4+ T cell counts, while 65 patients had increasing and 8 patients had decreasing absolute CD4+ T cell counts. In patients with plateaued CD4+ T cell counts, variability in absolute CD4+ T cell counts was greater than in percent CD4+ T cells (median coefficient of variation (CV) 16.6% [IQR 13.8-20.1%] and CV 9.6% [IQR 7.4-13.0%], respectively). Patients with increasing CD4+ T cell counts had greater variability in absolute CD4+ T cell counts than those with plateaued CD4 T cell counts (CV 19.5% [IQR 16.1-23.8%], p<0.001) while there was no difference in percent CD4+ T cell variability between the two groups. As previously reported, untreated patients had CVs significantly higher than patients on cART (CVs of 21.1% [IQR 17.2-32.0%], p<0.001 and 15.2% (IQR 10.7-20.0%), p<0.001, respectively). Age or sex did not affect the degree of CD4+ variation. Conclusions Adults with stable, virologically-suppressed HIV infection continue to have significant variations in individual absolute CD4+ T cell and percent CD4+ T cell counts; this variation can be of clinical relevance especially around CD4+ thresholds. However, the variation seen in individuals on cART is substantially less than in untreated subjects. PMID:26110761

  4. Cerebrospinal fluid B cells from multiple sclerosis patients are subject to normal germinal center selection

    Microsoft Academic Search

    Christopher Harp; Jane Lee; Doris Lambracht-Washington; Elizabeth Cameron; Gregory Olsen; Elliot Frohman; Michael Racke; Nancy Monson

    2007-01-01

    Previous findings from our laboratory demonstrated that some clonally expanded cerebrospinal fluid (CSF) B cells from MS patients exhibit diminished mutation targeting patterns in comparison to typical B cells selected in the context of germinal centers (GCs). In order to determine whether the overall CSF B cell repertoires adhered to mutation patterns typical of GC-selected B cells, we analyzed the

  5. White blood cell counts, leukocyte ratios, and eosinophils as inflammatory markers in patients with coronary artery disease.

    PubMed

    Kounis, Nicholas G; Soufras, George D; Tsigkas, Grigorios; Hahalis, George

    2015-03-01

    Inflammation is a key feature of atherosclerosis and its clinical manifestations. The leukocyte count has emerged as a marker of inflammation that is widely available in clinical practice. Since inflammation plays a key role in atherosclerosis and its end results, discovering new biomarkers of inflammation becomes important in order to help diagnostic accuracy and provide prognostic information about coronary cardiac disease. In acute coronary syndromes and percutaneous coronary intervention, elevated levels of almost all subtypes of white blood cell counts, including eosinophils, monocytes, neutrophils, and lymphocytes, and neutrophil-lymphocyte ratio and eosinophil-leukocyte ratio constitute independent predictors of adverse outcomes. Eosinophil count and eosinophil-leukocyte ratio, in particular, emerge as novel biomarkers for risk stratification in patients with coronary artery disease. Since the presence of eosinophils denotes hypersensitivity inflammation and hypersensitivity associated with Kounis syndrome, this reality is essential for elucidating the etiology of inflammation in order to consider predictive and preventive measures and to apply the appropriate therapeutic methods. PMID:24770327

  6. Counting Quail

    E-print Network

    Rollins, Dale; Brooks, Jason; Wilkins, Neal; Ransom, Dean

    2005-10-05

    at least three counts. Helicopter surveys Aerial surveys (conducted from a helicopter or airplane) are commonly used to count deer and pronghorn antelope in Texas, and have been used recently to survey quail. Generally, a quail survey is conducted...

  7. The Risk of Metabolic Syndrome According to the White Blood Cell Count in Apparently Healthy Korean Adults

    PubMed Central

    Jung, Chan-Hee; Kim, Bo-Yeon; Park, Se Eun; Rhee, Eun-Jung; Park, Cheol-Young; Oh, Ki-Won; Mok, Ji-Oh; Kim, Chul-Hee; Park, Sung-Woo; Kim, Sun-Woo; Kang, Sung-Koo

    2013-01-01

    Purpose Considerable amount of interest has been focused on the positive relationship between inflammation and the metabolic syndrome (MS). However, few studies have been performed to evaluate the relationship between baseline white blood cell (WBC) count and future risk for developing MS. Therefore, we investigated whether the baseline plasma levels of WBC count could be associated with future risk for MS in apparently healthy Korean. Materials and Methods A total of 1135 subjects (781 men and 354 women with a mean age of 49 years), who underwent health examinations at Kangbuk Samsung Hospital in both 2002 and 2005 were enrolled. The presence of MS was defined using the modified criteria of the National Cholesterol Education Program Adult Treatment Panel III using BMI instead of waist circumference. Results The baseline levels of WBC count were significantly higher among incident MS cases than among subjects without MS. The relative risks of incident MS were 1.4, 3.2 and 2.7 for WBC quartiles 2, 3, and 4, respectively, when compared with the first quartile (p-value for trend <0.001). These positive associations persisted after adjustment for baseline body mass index, blood pressure, fasting glucose, high density lipoprotein-cholesterol, triglyceride and homeostatic model assessment-insulin resistance; adjusted relative risk of incident MS for the 2nd, 3rd and 4th quartile groups vs. the lowest quartile were 1.2, 2.4 and 1.7, respectively (p-value for trend =0.011). Conclusion This retrospective cohort study suggests that an elevated WBC count could be associated with incident MS, suggesting that baseline inflammation mirrored by WBC level can impact future MS development. PMID:23549805

  8. Mobilization and collection of peripheral blood stem cells: guidelines for blood volume to process, based on CD34-positive blood cell count in adults and children.

    PubMed

    Anguita-Compagnon, A T; Dibarrart, M T; Palma, J; Paredes, L; Mosso, C; Montalva, R; Salas, L; Araos, D; Delgado, I; Majlis, A

    2010-01-01

    We report 189 mobilizations and 489 collections of peripheral blood stem cells (PBSC) performed in 139 autologous transplantation patients and in 28 donors for allogeneic transplantations whose ages ranged from 2-68 years. We observed a correlation (P < .001; Pearson's coefficient 0.64) between CD34-positive cells and granulocyte-macrophage colony-forming units examined to estimate PBSC. In a subset of 287 collections (97 adults and 49 children) we obtained peripheral blood (PB) CD34-positive cell counts at 2 to 4 hours before leukapheresis. We noted a correlation between PB CD34-positive cell counts before leukapheresis and the number of CD34-positive cells per kilogram of body weight collected in the whole apheresis of the day (P < .001; Pearson's coefficient 0.82). An even better correlation was obtained between PB CD34-positive cells preapheresis and the yield of each individual blood volume (BV) processed (P < .001; Pearson's coefficient 0.87). Healthy donors and patients in each age group behaved similarly. In addition, the collection yield was greater among children than adults. These findings allowed us to develop a simple predictive model to estimate the BV to process for a target dose of CD34-positive cells per kilogram, based on the level of PBSC before apheresis in children and adults. PMID:20172346

  9. Let's Count!

    NSDL National Science Digital Library

    Ms. Popwell

    2010-09-20

    Let's practice our counting skills with these fun games! Let's soar into the sky and practice Counting on a Cloud! The ants need lining up, let's Count the Ants! Help Rabbit eat his carrots by dropping the correct number of food into the basket! ...

  10. Satellited 4q identified in amniotic fluid cells

    SciTech Connect

    Miller, I.; Hsieh, C.L.; Songster, G. [Stanford Univ. Medical Center, Stanford, CA (United States)] [and others

    1995-01-16

    Extra material was identified on the distal long arm of a chromosome 4 in an amniotic fluid specimen sampled at 16.6 weeks of gestational age. There was no visible loss of material from chromosome 4, and no evidence for a balanced rearrangement. The primary counseling issue in this case was advanced maternal age. Ultrasound findings were normal, and family history was unremarkable. The identical 4qs chromosome was observed in cells from a paternal peripheral blood specimen and appeared to be an unbalanced rearrangement. This extra material was NOR positive in lymphocytes from the father, but was negative in the fetal amniocytes. Father`s relatives were studied to verify the familial origin of this anomaly. In situ hybridization with both exon and intron sequences of ribosomal DNA demonstrated that ribosomal DNA is present at the terminus of the 4qs chromosome in the fetus, father, and paternal grandmother. This satellited 4q might have been derived from a translocation event that resulted in very little or no loss from the 4q and no specific phenotype. This derivative chromosome 4 has been inherited through at least 3 generations of phenotypically normal individuals. 8 refs., 3 figs.

  11. Computational fluid dynamics modeling of proton exchange membrane fuel cells

    SciTech Connect

    UM,SUKKEE; WANG,C.Y.; CHEN,KEN S.

    2000-02-11

    A transient, multi-dimensional model has been developed to simulate proton exchange membrane (PEM) fuel cells. The model accounts simultaneously for electrochemical kinetics, current distribution, hydrodynamics and multi-component transport. A single set of conservation equations valid for flow channels, gas-diffusion electrodes, catalyst layers and the membrane region are developed and numerically solved using a finite-volume-based computational fluid dynamics (CFD) technique. The numerical model is validated against published experimental data with good agreement. Subsequently, the model is applied to explore hydrogen dilution effects in the anode feed. The predicted polarization cubes under hydrogen dilution conditions are found to be in qualitative agreement with recent experiments reported in the literature. The detailed two-dimensional electrochemical and flow/transport simulations further reveal that in the presence of hydrogen dilution in the fuel stream, hydrogen is depleted at the reaction surface resulting in substantial kinetic polarization and hence a lower current density that is limited by hydrogen transport from the fuel stream to the reaction site.

  12. Increased CD133+ cell infiltration in the rat brain following fluid percussion injury?

    PubMed Central

    Wei, Ming; Zhou, Ziwei; Li, Shenghui; Jing, Chengwei; Zhi, Dashi; Zhang, Jianning

    2012-01-01

    The prominin-1/CD133 epitope is expressed in undifferentiated cells. Studies have reported that craniocerebral trauma in animal models of fluid percussion injury induces production of a specific stem cell subgroup. It has been hypothesized that fluid percussion injury induces CD133+ cell infiltration in the brain tissue. The present study established a traumatic brain injury model through fluid percussion injury. Immunohistochemical staining showed significantly increased CD133 antigen expression in the rat brain following injury. CD133+ cells were mainly distributed in hippocampal CA1–3 regions, as well as the dentate gyrus and hilus, of the lesioned hemisphere. Occasional cells were also detected in the cortex. In addition, reverse transcription-PCR revealed that no change in CD133 mRNA expression in injured brain tissue. These results suggested that fluid percussion injury induced CD133 antigen expression in the brain tissues as a result of conformational epitope changes, but not transcriptional expression. PMID:25806069

  13. Count Around

    NSDL National Science Digital Library

    2011-08-23

    Learners explore their surroundings while reasoning about categories and counting. Pose a question that involves locating items in the room or building, and have learners count how many they can find—and figure out "what counts." It’s easy to vary the question for different levels of challenge. For instance, for less challenge, ask: How many light switches are in the room? For more, ask: How many light sources are in the room? Once everyone has counted, engage the group in discussing findings: Why might the answers differ even if everyone counted correctly? Available as a web page or downloadable pdf. Students should be able to write the numbers to 12.

  14. The Fluid-Kinetic Particle-in-Cell Solver for Plasma Simulations

    E-print Network

    Markidis, Stefano; Lapenta, Giovanni; Ronnmark, Kjell; Hamrin, Maria; Meliani, Zakaria; Laure, Erwin

    2013-01-01

    A new method that solves concurrently the multi-fluid and Maxwell's equations has been developed for plasma simulations. By calculating the stress tensor in the multi-fluid momentum equation by means of computational particles moving in a self-consistent electromagnetic field, the kinetic effects are retained while solving the multi-fluid equations. The Maxwell's and multi-fluid equations are discretized implicitly in time enabling kinetic simulations over time scales typical of the fluid simulations. The fluid-kinetic Particle-in-Cell solver has been implemented in a three-dimensional electromagnetic code, and tested against the ion cyclotron resonance and magnetic reconnection problems. The new method is a promising approach for coupling fluid and kinetic methods in a unified framework.

  15. The Time Series Image Analysis of the HeLa Cell Using Viscous Fluid Registration

    Microsoft Academic Search

    Soichiro Tokuhisa; Kunihiko Kaneko

    2010-01-01

    \\u000a Optical microscopy image analysis is important in the life science research. To obtain the motion of the cell, we use the\\u000a viscous fluid registration method based on fluid dynamics. Viscous fluid registration deforms an image at time t to the next image at time t+1. In this algorithm, there is a problem that an object cannot be divided into two.

  16. Enhanced generation of retinal progenitor cells from human retinal pigment epithelial cells induced by amniotic fluid

    PubMed Central

    2012-01-01

    Background Retinal progenitor cells are a convenient source of cell replacement therapy in retinal degenerative disorders. The purpose of this study was to evaluate the expression patterns of the homeobox genes PAX6 and CHX10 (retinal progenitor markers) during treatment of human retinal pigment epithelium (RPE) cells with amniotic fluid (AF), RPE cells harvested from neonatal cadaver globes were cultured in a mixture of DMEM and Ham's F12 supplemented with 10% FBS. At different passages, cells were trypsinized and co-cultured with 30% AF obtained from normal fetuses of 1416 weeks gestational age. Results Compared to FBS-treated controls, AF-treated cultures exhibited special morphological changes in culture, including appearance of spheroid colonies, improved initial cell adhesion and ordered cell alignment. Cell proliferation assays indicated a remarkable increase in the proliferation rate of RPE cells cultivated in 30% AF-supplemented medium, compared with those grown in the absence of AF. Immunocytochemical analyses exhibited nuclear localization of retinal progenitor markers at a ratio of 33% and 27% for CHX10 and PAX6, respectively. This indicated a 3-fold increase in retinal progenitor markers in AF-treated cultures compared to FBS-treated controls. Real-time PCR data of retinal progenitor genes (PAX6, CHX10 and VSX-1) confirmed these results and demonstrated AF's capacity for promoting retinal progenitor cell generation. Conclusion Taken together, the results suggest that AF significantly promotes the rate of retinal progenitor cell generation, indicating that AF can be used as an enriched supplement for serum-free media used for the in vitro propagation of human progenitor cells. PMID:22490806

  17. Somatic Cell Count, Electrical Conductivity, and Serum Albumin Concentration for Detecting Bovine Mastitis

    Microsoft Academic Search

    R. F. Sheldrake; G. D. McGregor; R. J. T. Hoare

    1983-01-01

    Cell concentration, electrical conduc- tivity, and serum albumin concentration of milk were evaluated for predicting infection status of quarters in three herds. Probability of misclassifying quarters was lowest for cell concentration. For discriminating quarters infected with Staphylococcus aureus from quarters free from infection, probability of misclassifi- cation for cell concentration ranged from 8 to 20% among herds. For electrical conductivity

  18. Responses of Escherichia coli, Listeria monocytogenes, and Staphylococcus aureus to Simulated Food Processing Treatments, Determined Using Fluorescence-Activated Cell Sorting and Plate Counting?

    PubMed Central

    Kennedy, Deirdre; Cronin, Ultan P.; Wilkinson, Martin G.

    2011-01-01

    Three common food pathogenic microorganisms were exposed to treatments simulating those used in food processing. Treated cell suspensions were then analyzed for reduction in growth by plate counting. Flow cytometry (FCM) and fluorescence-activated cell sorting (FACS) were carried out on treated cells stained for membrane integrity (Syto 9/propidium iodide) or the presence of membrane potential [DiOC2(3)]. For each microbial species, representative cells from various subpopulations detected by FCM were sorted onto selective and nonselective agar and evaluated for growth and recovery rates. In general, treatments giving rise to the highest reductions in counts also had the greatest effects on cell membrane integrity and membrane potential. Overall, treatments that impacted cell membrane permeability did not necessarily have a comparable effect on membrane potential. In addition, some bacterial species with extensively damaged membranes, as detected by FCM, appeared to be able to replicate and grow after sorting. Growth of sorted cells from various subpopulations was not always reflected in plate counts, and in some cases the staining protocol may have rendered cells unculturable. Optimized FCM protocols generated a greater insight into the extent of the heterogeneous bacterial population responses to food control measures than did plate counts. This study underlined the requirement to use FACS to relate various cytometric profiles generated by various staining protocols with the ability of cells to grow on microbial agar plates. Such information is a prerequisite for more-widespread adoption of FCM as a routine microbiological analytical technique. PMID:21602370

  19. Microfluidics-based in situ padlock/rolling circle amplification system for counting single DNA molecules in a cell.

    PubMed

    Kuroda, Arisa; Ishigaki, Yuri; Nilsson, Mats; Sato, Kiichi; Sato, Kae

    2014-01-01

    In situ padlock/rolling circle amplification (RCA) is a method used to amplify, visualize, and quantify target DNA molecules in cells. However, the multiple reaction steps involved make this technique costly and cumbersome. We developed a novel, simplified, automated microfluidic system for RCA, and demonstrated its effectiveness by counting amplified mitochondrial DNA fragments in HeLa cells. After optimizing the volume of the reaction solutions and washing buffer composition, the product yield was equal to that obtained by the conventional manual method. The required volume of reagents was reduced to 10 ?L, which is less than half the volume used in the conventional method. To the best of our knowledge, this is the first report of an automated microfluidic method for in situ padlock/RCA, which can be useful for making highly efficient pathological diagnoses. PMID:25492458

  20. Response of Listeria monocytogenes to Disinfection Stress at the Single-Cell and Population Levels as Monitored by Intracellular pH Measurements and Viable-Cell Counts?

    PubMed Central

    Kastbjerg, Vicky G.; Nielsen, Dennis S.; Arneborg, Nils; Gram, Lone

    2009-01-01

    Listeria monocytogenes has a remarkable ability to survive and persist in food production environments. The purpose of the present study was to determine if cells in a population of L. monocytogenes differ in sensitivity to disinfection agents as this could be a factor explaining persistence of the bacterium. In situ analyses of Listeria monocytogenes single cells were performed during exposure to different concentrations of the disinfectant Incimaxx DES to study a possible population subdivision. Bacterial survival was quantified with plate counting and disinfection stress at the single-cell level by measuring intracellular pH (pHi) over time by fluorescence ratio imaging microscopy. pHi values were initially 7 to 7.5 and decreased in both attached and planktonic L. monocytogenes cells during exposure to sublethal and lethal concentrations of Incimaxx DES. The response of the bacterial population was homogenous; hence, subpopulations were not detected. However, pregrowth with NaCl protected the planktonic bacterial cells during disinfection with Incimaxx (0.0015%) since pHi was higher (6 to 6.5) for the bacterial population pregrown with NaCl than for cells grown without NaCl (pHi 5 to 5.5) (P < 0.05). The protective effect of NaCl was reflected by viable-cell counts at a higher concentration of Incimaxx (0.0031%), where the salt-grown population survived better than the population grown without NaCl (P < 0.05). NaCl protected attached cells through drying but not during disinfection. This study indicates that a population of L. monocytogenes cells, whether planktonic or attached, is homogenous with respect to sensitivity to an acidic disinfectant studied on the single-cell level. Hence a major subpopulation more tolerant to disinfectants, and hence more persistent, does not appear to be present. PMID:19411424

  1. Single cell rheometry with a microfluidic constriction: Quantitative control of friction and fluid leaks between cell and channel walls

    E-print Network

    Bico,José

    leaks between cell and channel walls Pascal Preira, Marie-Pierre Valignat, José Bico, and Olivier leaks between cell and channel walls Pascal Preira,1 Marie-Pierre Valignat,1 Jose Bico,2 and Olivier the deformation of a cell in a microfluidic constriction, provided that friction and fluid leaks effects between

  2. Cell growth-promoting activity of fluid from eye sacs of the bubble-eye goldfish (Carassius auratus).

    PubMed

    Sawatari, Etsuko; Hashimoto, Hisashi; Matsumura, Takaharu; Iwata, Yasuhiro; Yamamoto, Naoki; Yokoyama, Yoshihiro; Wakamatsu, Yuko

    2009-04-01

    The growth-promoting effects of fish body fluids, such as serum and embryonic extract, on fish cell cultures have been widely demonstrated. The bubble-eye variety of aquarium goldfish is characterized as having a large sac filled with fluid (sac fluid) under each eye. These sacs are believed to contain lymph, which is similar in composition to serum or blood plasma. In order to test whether the sac fluid can be used as an additive for fetal bovine serum (FBS) in growth factor supplements, we compared cell growth in media containing FBS together with different concentrations of sac fluid. A dose-dependent growth-promotion effect was observed in early passage caudal fin cells from both medaka and zebrafish. Cell-growth promotion was also confirmed in early passage medaka blastula cells and in a zebrafish embryonic cell line (ZF4). Replacement of the fluid in the eye sacs of bubble-eyes occurs within a couple of months after the sac fluid has been harvested, and the cell-growth promoting activity of the new fluid is similar to that of the fluid that was tapped initially. These findings suggest that sac fluid can be used as a growth-promoting supplement for fish cell culture. Importantly, the ability of the goldfish to replace the fluid combined with the fact that equipotent fluid can be repeatedly harvested from the eye sacs means that a sustainable source of the fluid can be obtained without needing to sacrifice the fish. PMID:19798918

  3. Impact on life expectancy of HIV-1 positive individuals of CD4+ cell count and viral load response to antiretroviral therapy

    PubMed Central

    May, Margaret T.; Gompels, Mark; Delpech, Valerie; Porter, Kholoud; Orkin, Chloe; Kegg, Stephen; Hay, Phillip; Johnson, Margaret; Palfreeman, Adrian; Gilson, Richard; Chadwick, David; Martin, Fabiola; Hill, Teresa; Walsh, John; Post, Frank; Fisher, Martin; Ainsworth, Jonathan; Jose, Sophie; Leen, Clifford; Nelson, Mark; Anderson, Jane; Sabin, Caroline

    2014-01-01

    Objective: The objective of this study is to estimate life expectancies of HIV-positive patients conditional on response to antiretroviral therapy (ART). Methods: Patients aged more than 20 years who started ART during 2000–2010 (excluding IDU) in HIV clinics contributing to the UK CHIC Study were followed for mortality until 2012. We determined the latest CD4+ cell count and viral load before ART and in each of years 1–5 of ART. For each duration of ART, life tables based on estimated mortality rates by sex, age, latest CD4+ cell count and viral suppression (HIV-1 RNA <400?copies/ml), were used to estimate expected age at death for ages 20–85 years. Results: Of 21?388 patients who started ART, 961 (4.5%) died during 110?697 person-years. At start of ART, expected age at death [95% confidence interval (CI)] of 35-year-old men with CD4+ cell count less than 200, 200–349, at least 350?cells/?l was 71 (68–73), 78 (74–82) and 77 (72–81) years, respectively, compared with 78 years for men in the general UK population. Thirty-five-year-old men who increased their CD4+ cell count in the first year of ART from less than 200 to 200–349 or at least 350?cells/?l and achieved viral suppression gained 7 and 10 years, respectively. After 5 years on ART, expected age at death of 35-year-old men varied from 54 (48–61) (CD4+ cell count <200?cells/?l and no viral suppression) to 80 (76–83) years (CD4+ cell count ?350?cells/?l and viral suppression). Conclusion: Successfully treated HIV-positive individuals have a normal life expectancy. Patients who started ART with a low CD4+ cell count significantly improve their life expectancy if they have a good CD4+ cell count response and undetectable viral load. PMID:24556869

  4. Validation of World Health Organisation HIV\\/AIDS Clinical Staging in Predicting Initiation of Antiretroviral Therapy and Clinical Predictors of Low CD4 Cell Count in Uganda

    Microsoft Academic Search

    Steven Baveewo; Francis Ssali; Charles Karamagi; Joan N. Kalyango; Judith A. Hahn; Kenneth Ekoru; Peter Mugyenyi; Elly Katabira; Malcolm Gracie Semple

    2011-01-01

    IntroductionThe WHO clinical guidelines for HIV\\/AIDS are widely used in resource limited settings to represent the gold standard of CD4 counts for antiviral therapy initiation. The utility of the WHO-defined stage 1 and 2 clinical factors used in WHO HIV\\/AIDS clinical staging in predicting low CD4 cell count has not been established in Uganda. Although the WHO staging has shown

  5. arXiv:astro-ph/0401559v126Jan2004 Self-Calibration of Cluster Dark Energy Studies: Counts in Cells

    E-print Network

    Hu, Wayne

    arXiv:astro-ph/0401559v126Jan2004 Self-Calibration of Cluster Dark Energy Studies: Counts in Cells of Chicago, Chicago IL 60637 Cluster number counts can constrain the properties of dark energy if and only constraints on the dark energy equation of state by a factor of 2 or more to (w) = 0.06 for a deep 4000 deg2

  6. Bulk tank milk somatic cell counts in dairy herds with different bovine viral diarrhoea virus status in Poland.

    PubMed

    Rola, Jolanta G; Larska, Magdalena; Grzeszuk, Monika; Bocian, Lukasz; Kuta, Aleksandra; Polak, Miroslaw P; Rola, Jerzy

    2014-09-01

    The aim of the study was to examine the effect of bovine viral diarrhoea virus (BVDV) infection on bulk tank milk somatic cell counts (BMSCC). Twenty nine dairy farms supplying milk to a dairy in Eastern Poland were recruited for the study. Bulk milk ELISA and RT-PCR were used to determine the BVDV infection status and the presence of PI animals in the farms. The BMSCC mean values for the BVDV seronegative (218.7 × 10(3)cells/ml; SD: 89.8) and seropositive (214.9 × 10(3)cells/ml; SD: 74.0) herds did not differ significantly. To assess the relationship between BVDV infection and BMSCC a multilevel mixed-effects linear model was used. No statistically significant effect of BVDV infection on BMSCC was found. The mean values of BMSCC for the herds with PI individuals measured before (230.1 × 10(3)cells/ml, SD: 64.9) and after (223.3 × 10(3)cells/ml, SD: 62.4) the PI removal were not statistically different. An increase in herd size was associated with a significant decrease in BMSCC. An increase in BMSCC was observed during summer (from May to September) compared to during winter (from October to April). PMID:25023907

  7. A FLUID-CELL INTERACTION AND ADHESION ALGORITHM FOR TISSUE-COATING OF CARDIOVASCULAR IMPLANTS

    E-print Network

    Canic, Suncica

    A FLUID-CELL INTERACTION AND ADHESION ALGORITHM FOR TISSUE-COATING OF CARDIOVASCULAR IMPLANTS JIAN and adhesion algorithm applied to modeling the cell coating of artificial surfaces of cardiovascular implants describing cell coating of cardiovascular implants are presented in Section 2. To study this problem we

  8. Demonstration of astrocytes in cultured amniotic fluid cells of three cases with neural-tube defect

    Microsoft Academic Search

    Marion Cremer; Melitta Schachner; Thomas Cremer; Werner Schmidt; Theda Voigtlfinder

    1981-01-01

    We have investigated the origin of rapidly adhering (RA) cells in three cases of neural tube defects (two anencephali, one encephalocele). We were able to demonstrate the presence of glial fibrillary acidic (GFA) protein in variable percentages (4–80%) of RA cells cultured for 4–6 days by use of indirect immunofluorescence with GFA antiserum. Cells cultured from amniotic fluids of normal

  9. The FASEB Journal Research Communication Fluid shear stress primes mouse embryonic stem cells

    E-print Network

    Voldman, Joel

    The FASEB Journal · Research Communication Fluid shear stress primes mouse embryonic stem cells stress is a ubiquitous environmen- tal cue experienced by stem cells when they are being differentiated or expanded in perfusion cultures. How- ever, its role in modulating self-renewing stem cell phenotypes

  10. FluidNet: A Flexible Cloud-based Radio Access Network for Small Cells

    E-print Network

    Krishnamurthy, Srikanth

    FluidNet: A Flexible Cloud-based Radio Access Network for Small Cells Karthikeyan Sundaresan NEC-RAN) have been proposed as a cost-efficient way of deploying small cells. Unlike conven- tional RANs, a C operation of BBUs and scalable deployment of light-weight RRHs as small cells. In this work, we argue

  11. Effect of somatic cell count in goat milk on yield, sensory quality and fatty acid profile of semi-hard cheese

    Technology Transfer Automated Retrieval System (TEKTRAN)

    This study investigated the effect of somatic cell count (SCC) of goat milk on yield, free fatty acid (FFA) profile, and sensory quality of semi-hard cheese. Thirty kilograms of goat milk with mean SCC levels of 410,000 (Low), 770,000 (Medium), and 1,250,000 cells/mL (High) was obtained for the manu...

  12. CYTOLOGICAL QUALITY OF GOAT MILK ON THE BASIS OF THE SOMATIC CELL COUNT JAKO?C CYTOLOGICZNA MLEKA KÓZ NA PODSTAWIE ZAWARTO?CI KOMÓREK SOMATYCZNYCH

    Microsoft Academic Search

    Henryka BERNACKA

    The aim of the present paper was to evaluate the cytological quality of goat milk based on the somatic cell count in respective months of lactation. Besides there was defined the effect of somatic cell on the milk production and chemical composition of milk. The research covered goats of color improved breed in the 2nd and 3rd lactation. Daily milk

  13. Supplemental Figure 1: Pil1 and Lsp1 behave identically during de novo eisosome formation (a) Number of eisosomes where counted in cells expressing either Lsp1-GFP,

    E-print Network

    Walter, Peter

    Supplemental Figure 1: Pil1 and Lsp1 behave identically during de novo eisosome formation (a) Number of eisosomes where counted in cells expressing either Lsp1-GFP, TWY113 (red) or Pil1-GFP, TWY110 (green) and are shown as a function of the bud cell surface. (b) Eisosome deposition as visualized by Lsp

  14. Comparison of particle in cell and fluid models in plasma display panels simulation

    NASA Astrophysics Data System (ADS)

    Benstâali, W.; Belasri, A.

    2013-03-01

    In this paper, a particle in cell model (PIC) was developed. We compare the major characteristics of a single discharge obtained from our PIC model and the fluid model. Fluid model uses different approximations as Local field (LFA). The particle in cell (PIC) model, although its costs in time, still an efficient tool to understand a PDP discharge. Even there is some differences as the delay time and the ionization, the discharge still be correctly described as the energy balance.

  15. Photon counting.

    PubMed

    Morton, G A

    1968-01-01

    The fundamentals of photon counting using photomultipliers are described, including criteria for selecting suitable photomultipliers, some of the precautions that must be taken in using these devices, and methods of calculating the counting errors that may occur under various conditions of measurement. Problems of determining the time distribution of photons and, in particular, the coincident emission of photons which may be encountered in lasers and other simulated emission sources are also discussed. The question of photon counting with photoconductors is reviewed, and it is shown that it is extremely difficult, if not impossible, to achieve photon counting with simple photoconductors. However, carrier multiplication with photoconductive multipliers should eventually make possible photon counting with photoconductors. Photoconductive multipliers in one form or another have high quantum efficiency and wide spectral response, and will almost inevitably replace photomultipliers for photon counting. PMID:20062394

  16. Method for filling the cavities of cells with a chromogenic fluid

    DOEpatents

    Tonazzi, J.C.L.; Kucharczyk, J.E. Jr.; Agrawal, A.

    1999-01-05

    A method and apparatus are disclosed for filling a cell cavity positioned between a first substrate and a second substrate with a cell filling liquid. The method entails forming at least one evacuation cavity encompassing at least a portion of an outer surface of each of the first and second substrates of a cell containing a cell cavity and isolating the cell cavity from the evacuation cavity; reducing a pressure in each of the evacuation cavity and the cell cavity; and dispensing the cell filling fluid into the cell cavity. The application is to the fabrication of electrochromic windows. 22 figs.

  17. Method for filling the cavities of cells with a chromogenic fluid

    DOEpatents

    Tonazzi, Juan C. Lopez (Tucson, AZ); Kucharczyk, Jr., Joseph E. (Tucson, AZ); Agrawal, Anoop (Tucson, AZ)

    1999-01-01

    A method and apparatus are disclosed for filling a cell cavity positioned between a first substrate and a second substrate with a cell filling liquid. The method entails forming at least one evacuation cavity encompassing at least a portion of an outer surface of each of the first and second substrates of a cell containing a cell cavity and isolating the cell cavity from the evacuation cavity; reducing a pressure in each of the evacuation cavity and the cell cavity; and dispensing the cell filling fluid into the cell cavity.

  18. Infused peripheral blood autograft absolute lymphocyte count correlates with day 15 absolute lymphocyte count and clinical outcome after autologous peripheral hematopoietic stem cell transplantation in non-Hodgkin's lymphoma

    Microsoft Academic Search

    L F Porrata; M R Litzow; D J Inwards; D A Gastineau; S B Moore; A A Pineda; K L Bundy; D J Padley; D Persky; S M Ansell; I N M Micallef; S N Markovic

    2004-01-01

    Absolute lymphocyte count at day 15 (ALC-15) after autologous peripheral blood hematopoietic stem cell transplantation (APHSCT) is an independent prognostic factor for survival in non-Hodgkin's lymphoma (NHL). Factors affecting ALC-15 remain unknown. We hypothesized that dose of infused autograft lymphocytes (A-ALC) directly impacts upon ALC-15. A total of 190 consecutive NHL patients received A-ALC between 1993 and 2001. The primary

  19. Somatic cell counts of milk from Dairy Herd Improvement herds during 2009

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Test-day data from all herds enrolled in Dairy Herd Improvement (DHI) somatic cell testing during 2009 were examined to assess the status of national milk quality. Cows with records failing some AIPL editing procedures were excluded. Somatic cell score (SCS) is reported to AIPL and was converted to ...

  20. Somatic Cell Counts of Milk from Dairy Herd Improvement Herds during 2007

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Test-day data from all herds enrolled in Dairy Herd Improvement (DHI) somatic cell testing during 2007 were examined to assess the status of national milk quality. Cows with records failing some AIPL editing procedures were excluded. Somatic cell score (SCS) is reported to AIPL and was converted to ...

  1. Somatic cell counts of milk from Dairy Herd Improvement herds during 2008

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Test-day data from all herds enrolled in Dairy Herd Improvement (DHI) somatic cell testing during 2008 were examined to assess the status of national milk quality. Cows with records failing some AIPL editing procedures were excluded. Somatic cell score (SCS) is reported to AIPL and was converted to ...

  2. Induction of E-cadherin+ human amniotic fluid cell differentiation into oocyte-like cells via culture in medium supplemented with follicular fluid

    PubMed Central

    LIU, TE; HUANG, YONGYI; BU, YANZHEN; ZHAO, YANHUI; ZOU, GANG; LIU, ZHIXUE

    2014-01-01

    Pluripotent human amniotic fluid cells (HuAFCs) can differentiate into various types of somatic cell in vitro. However, their differentiation into oocyte-like cells has never been described to the best of our knowledge. In the present study, differentiation of E-cadherin+ and E-cadherin? HuAFC sub-populations into oocyte-like cells was induced via culture in medium containing bovine follicular fluid and ?-mercaptoethanol. The E-cadherin+ HuAFCs expressed DAZL highly. Post-induction, cells with an oocyte-like phenotype were found among the E-cadherin+ HuAFCs, expressing markers specific to germ cells and oocytes (VASA, ZP3 and GDF9) and meiosis (DMC1 and SCP3). When specific small interfering RNA (siRNA) was used to suppress E-cadherin in the E-cadherin+ HuAFCs, the levels of DAZL expression were reduced. Post-induction, the morphology of the siRNA-E-cadherin HuAFCs was poorer and the expression levels of germ cell-specific markers were lower compared with those of the siRNA-mock HuAFCs. Therefore, E-cadherin+ HuAFCs could be more easily induced to differentiate into oocyte-like cells by bovine follicular fluid and ?-mercaptoethanol. In addition, the E-cadherin+ HuAFCs exhibited potential characteristics of DAZL protein expression, and thus it was conjectured that bovine follicular fluid acts on DAZL protein and promotes E-cadherin+ HuAFC differentiation into oocyte-like cells. PMID:24788191

  3. Induction of E-cadherin+ human amniotic fluid cell differentiation into oocyte-like cells via culture in medium supplemented with follicular fluid.

    PubMed

    Liu, Te; Huang, Yongyi; Bu, Yanzhen; Zhao, Yanhui; Zou, Gang; Liu, Zhixue

    2014-07-01

    Pluripotent human amniotic fluid cells (HuAFCs) can differentiate into various types of somatic cell in vitro. However, their differentiation into oocyte-like cells has never been described to the best of our knowledge. In the present study, differentiation of E-cadherin+ and E-cadherin- HuAFC sub-populations into oocyte-like cells was induced via culture in medium containing bovine follicular fluid and ?-mercaptoethanol. The E-cadherin+ HuAFCs expressed DAZL highly. Post-induction, cells with an oocyte-like phenotype were found among the E-cadherin+ HuAFCs, expressing markers specific to germ cells and oocytes (VASA, ZP3 and GDF9) and meiosis (DMC1 and SCP3). When specific small interfering RNA (siRNA) was used to suppress E-cadherin in the E-cadherin+ HuAFCs, the levels of DAZL expression were reduced. Post-induction, the morphology of the siRNA?E?cadherin HuAFCs was poorer and the expression levels of germ cell-specific markers were lower compared with those of the siRNA-mock HuAFCs. Therefore, E-cadherin+ HuAFCs could be more easily induced to differentiate into oocyte-like cells by bovine follicular fluid and ?-mercaptoethanol. In addition, the E-cadherin+ HuAFCs exhibited potential characteristics of DAZL protein expression, and thus it was conjectured that bovine follicular fluid acts on DAZL protein and promotes E-cadherin+ HuAFC differentiation into oocyte-like cells. PMID:24788191

  4. Prognosis of patients treated with cART from 36 months after initiation, according to current and previous CD4 cell count and plasma HIV-1 RNA measurements

    PubMed Central

    2011-01-01

    Objectives CD4 cell count and plasma viral load are well known predictors of AIDS and mortality in HIV-1-infected patients treated with combination antiretroviral therapy (cART). This study investigated, in patients treated for at least 3 years, the respective prognostic importance of values measured at cART initiation, and 6 and 36 months later, for AIDS and death. Methods Patients from 15 HIV cohorts included in the ART Cohort Collaboration, aged at least 16 years, antiretroviral-naive when they started cART and followed for at least 36 months after start of cART were eligible. Results Among 14 208 patients, the median CD4 cell counts at 0, 6 and 36 months were 210, 320 and 450 cells/µl, respectively, and 78% of patients achieved viral load less than 500 copies/ml at 6 months. In models adjusted for characteristics at cART initiation and for values at all time points, values at 36 months were the strongest predictors of subsequent rates of AIDS and death. Although CD4 cell count and viral load at cART initiation were no longer prognostic of AIDS or of death after 36 months, viral load at 6 months and change in CD4 cell count from 6 to 36 months were prognostic for rates of AIDS from 36 months. Conclusions Although current values of CD4 cell count and HIV-1 RNA are the most important prognostic factors for subsequent AIDS and death rates in HIV-1-infected patients treated with cART, changes in CD4 cell count from 6 to 36 months and the value of 6-month HIV-1 RNA are also prognostic for AIDS. PMID:19779320

  5. Physical and chemical characteristics of yoghurt produced from whole milk with different levels of somatic cell counts.

    PubMed

    Hachana, Y; Paape, Max J

    2012-05-01

    For yoghurts were made from milk with different levels of somatic cell count (SCC) (low 95,000 cells ml(- 1), intermediate 398,000 cells ml(- 1) and high 1,150,000 cells ml(- 1)). Yoghurt samples were analysed for the degree of proteolysis, lipolysis (free fatty acid (FFA) content), acidity, pH and apparent viscosity on days 1, 14 and 28. The SCC had no significant effect (p>0.05) on either the acidity or the pH of the yoghurt after 1 day of cold storage. However, significant effects (p < 0.05) of SCC were observed after 14 and 28 days of storage. Yoghurt samples made from intermediate and high SCC milk showed higher viscosity (p < 0.05) and lower (p < 0.05) casein content on days 14 and 28 of cold storage than yoghurt made from low SCC milk. High FFA concentrations (p < 0.05) were observed only in yoghurt made from high SCC milk. High SCC in milk increased both proteolysis and lipolysis in yoghurt during storage. PMID:21999630

  6. Fluorescence photon migration techniques for the on-farm measurement of somatic cell count in fresh cow's milk

    NASA Astrophysics Data System (ADS)

    Khoo, Geoffrey; Kuennemeyer, Rainer; Claycomb, Rod W.

    2005-04-01

    Currently, the state of the art of mastitis detection in dairy cows is the laboratory-based measurement of somatic cell count (SCC), which is time consuming and expensive. Alternative, rapid, and reliable on-farm measurement methods are required for effective farm management. We have investigated whether fluorescence lifetime measurements can determine SCC in fresh, unprocessed milk. The method is based on the change in fluorescence lifetime of ethidium bromide when it binds to DNA from the somatic cells. Milk samples were obtained from a Fullwood Merlin Automated Milking System and analysed within a twenty-four hour period, over which the SCC does not change appreciably. For reference, the milk samples were also sent to a testing laboratory where the SCC was determined by traditional methods. The results show that we can quantify SCC using the fluorescence photon migration method from a lower bound of 4x105 cells mL-1 to an upper bound of 1 x 107 cells mL-1. The upper bound is due to the reference method used while the cause of the lower boundary is unknown, yet.

  7. Flow Cytometry Total Cell Counts: A Field Study Assessing Microbiological Water Quality and Growth in Unchlorinated Drinking Water Distribution Systems

    PubMed Central

    Liu, G.; Van der Mark, E. J.; Verberk, J. Q. J. C.; Van Dijk, J. C.

    2013-01-01

    The objective of this study was to evaluate the application of flow cytometry total cell counts (TCCs) as a parameter to assess microbial growth in drinking water distribution systems and to determine the relationships between different parameters describing the biostability of treated water. A one-year sampling program was carried out in two distribution systems in The Netherlands. Results demonstrated that, in both systems, the biomass differences measured by ATP were not significant. TCC differences were also not significant in treatment plant 1, but decreased slightly in treatment plant 2. TCC values were found to be higher at temperatures above 15°C than at temperatures below 15°C. The correlation study of parameters describing biostability found no relationship among TCC, heterotrophic plate counts, and Aeromonas. Also no relationship was found between TCC and ATP. Some correlation was found between the subgroup of high nucleic acid content bacteria and ATP (R2 = 0.63). Overall, the results demonstrated that TCC is a valuable parameter to assess the drinking water biological quality and regrowth; it can directly and sensitively quantify biomass, detect small changes, and can be used to determine the subgroup of active HNA bacteria that are related to ATP. PMID:23819117

  8. Genetic parameters for faecal egg count, packed-cell volume and body-weight in Santa Inês lambs

    PubMed Central

    2009-01-01

    Worm infection is one of the main factors responsible for economic losses in sheep breeding in Brazil. Random regression analysis was used to estimate genetic parameters for the factors faecal egg-count (FEC), packed-cell volume (PCV) and body weight (BW) in Santa Inês lambs. Data from 119 female, offspring of nine rams, were collected between December, 2005 and December, 2006, from the experimental flock of Embrapa Tabuleiros Costeiros, the Brazilian Agricultural Research Corporation located in Frei Paulo, SE, Brazil. After weaning, females were drenched until the faecal egg count had dropped to zero. Two natural challenges were undertaken. FEC heritability was extremely variable, this increasing from 0.04 to 0.27 in the first challenge and from 0.01 to 0.52 during the second. PCV heritability peaks were 0.31 and 0.12 in the first and second challenges, respectively. In the second challenge, BW heritability was close to 0.90. The genetic correlations among these traits did not differ from zero. There is the possibility of increasing parasite resistance in Santa Inês by selecting those animals with lower FEC. Selection to increase resistance will not adversely affect lamb-growth, although lambs with a slow growth-rate may be more susceptible to infection. PMID:21637682

  9. Different Immunological Phenotypes Associated with Preserved CD4+ T Cell Counts in HIV-Infected Controllers and Viremic Long Term Non-Progressors

    PubMed Central

    Gaardbo, Julie Christine; Hartling, Hans J.; Ronit, Andreas; Thorsteinsson, Kristina; Madsen, Hans Ole; Springborg, Karoline; Gjerdrum, Lise Mette Rahbek; Birch, Carsten; Laye, Matthew; Ullum, Henrik; Andersen, Åse Bengaard; Nielsen, Susanne Dam

    2013-01-01

    Background HIV-infected controllers control viral replication and maintain normal CD4+ T cell counts. Long Term Non-Progressors (LTNP) also maintain normal CD4+ T cell counts, but have on-going viral replication. We hypothesized that different immunological mechanisms are responsible for preserved CD4+ T cell counts in controllers and LTNP. Methods 25 HIV-infected controllers and 14 LTNP were included in this cross-sectional study. For comparison, 25 progressors and 34 healthy controls were included. Production and destruction of T cells were addressed by determination of T cell receptor excision circles (TREC), recent thymic emigrants, naïve cells, immune activation, senescence and apoptosis. Furthermore, telomere length was determined, and the amount of lymphoid tissue in tonsil biopsies was quantified. Results Controllers presented with partly preserved thymic output, preserved expression of the IL-7 receptor and IL-7 receptor density, and lower levels of destruction of cells than progressors resembling HIV-negative healthy controls. In contrast, LTNP appeared much like progressors, and different from controllers in immune activation, senescence, and apoptosis. Interestingly, CD8+ RTE, TREC and telomere length were partly preserved. Finally, both controllers and LTNP displayed decreased amounts of lymphoid tissue compared to healthy controls. Conclusions Controllers presented with an immunological profile different from LTNP. While controllers resembled healthy controls, LTNP were similar to progressors, suggesting different immunological mechanisms to be responsible for preserved CD4+ T cell counts in LTNP and controllers. However, both controllers and LTNP presented with reduced amounts of lymphoid tissue despite preserved CD4+ T cell counts, indicating HIV to cause damage even in non-progressors. PMID:23696852

  10. Single-molecule transcript counting of stem-cell markers in the mouse intestine

    E-print Network

    Itzkovitz, Shaul Shalev

    Determining the molecular identities of adult stem cells requires technologies for sensitive transcript detection in tissues. In mouse intestinal crypts, lineage-tracing studies indicated that different genes uniquely mark ...

  11. Variation in Somatic Cell Counts in Dairy Herd Improvement Milk Samples1

    Microsoft Academic Search

    G. W. Bodoh; W. J. Battista; L. H. Schultz; R. P. Johnston Jr.

    1976-01-01

    Information on variability of somatic cell numbers in Dairy Herd Improvement samples analyzed by the Filter-DNA method came from two field studies. Data consisted of 13,733 samples taken in 16 herds monthly for 2 yr in one sur- vey and 6285 observations in 134 herds each sampled once in the other. Mean cell numbers in the studies were 692,000 and

  12. Effect of exercise on erythrocyte count and blood activity concentration after /sup 99m/Tc in vivo red blood cell labeling

    SciTech Connect

    Konstam, M.A.; Tu'meh, S.; Wynne, J.; Beck, J.R.; Kozlowski, J.; Holman, B.L.

    1982-09-01

    We studied the effect of exercise on blood radiotracer concentration after /sup 99m/Tc in vivo red blood cell labeling. After red blood cell labeling, 13 subjects underwent maximal supine bicycle exercise. Radioactivity, analyzed with a well counter, was measured in heparinized venous blood samples drawn at rest and during peak exercise. Changes in activity were compared with changes in erythrocyte count. Activity and erythrocyte counts increased during exercise in all 13 subjects. Percent increase in activity correlated with percent increase in erythrocyte count (r . -0.78), but did not correlate with either duration of exercise or maximal heart rate. Twenty minutes after termination of exercise, activity and erythrocyte count had decreased from peak exercise values but remained higher than preexercise values. In nine nonexercised control subjects, samples drawn 20 minutes apart showed no change in activity or in erythrocyte count. We conclude that exercise increases blood activity, primarily because of an increase in erythrocyte count. During radionuclide ventriculography, blood activity must be measured before and after any intervention, particularly exercise, before a change in left ventricular activity can be attributed to a change in left ventricular volume.

  13. Human transgene-free amniotic-fluid-derived induced pluripotent stem cells for autologous cell therapy.

    PubMed

    Jiang, Guihua; Di Bernardo, Julie; Maiden, Michael M; Villa-Diaz, Luis G; Mabrouk, Omar S; Krebsbach, Paul H; O'Shea, K Sue; Kunisaki, Shaun M

    2014-11-01

    The establishment of a reliable prenatal source of autologous, transgene-free progenitor cells has enormous potential in the development of regenerative-medicine-based therapies for infants born with devastating birth defects. Here, we show that a largely CD117-negative population of human amniotic fluid mesenchymal stromal cells (AF-MSCs) obtained from fetuses with or without prenatally diagnosed anomalies are readily abundant and have limited baseline differentiation potential when compared with bone-marrow-derived MSCs and other somatic cell types. Nonetheless, the AF-MSCs could be easily reprogrammed into induced pluripotent stem cells (iPSCs) using nonintegrating Sendai viral vectors encoding for OCT4, SOX2, KLF4, and cMYC. The iPSCs were virtually indistinguishable from human embryonic stem cells in multiple assays and could be used to generate a relatively homogeneous population of neural progenitors, expressing PAX6, SOX2, SOX3, Musashi-1, and PSA-NCAM, for potential use in neurologic diseases. Further, these neural progenitors showed engraftment potential in vivo and were capable of differentiating into mature neurons and astrocytes in vitro. This study demonstrates the usefulness of AF-MSCs as an excellent source for the generation of human transgene-free iPSCs ideally suited for autologous perinatal regenerative medicine applications. PMID:25014361

  14. Counting Money

    NSDL National Science Digital Library

    Ms. Bunn

    2010-10-30

    Students will reinforce the idea of counting coins as well as adding different amounts of coins. First, play Shoot your fruit! to identify your numbers! Then, dive into Underwater Counting!! Ms. Eppes Class: First, visit farm stand to figure out how much it will cost to buy eggs and apples. Once you have completed the farm stand go on a spending spree! ...

  15. Associations between decreased fertility and management factors, claw health, and somatic cell count in Swedish dairy cows.

    PubMed

    Lomander, H; Svensson, C; Hallén-Sandgren, C; Gustafsson, H; Frössling, J

    2013-10-01

    The aim of this retrospective single-cohort study was to investigate if a rapid change in feeding, management, or housing or an increasing incidence of claw diseases or udder health problems is associated with decreased reproductive performance. Data on individual cows and herds were retrieved from the Swedish official milk recording system and questionnaire data on feeding system was obtained from the regional dairy associations. In total, 63,561 cows in 759 herds were included in the study. The associations between the probability of pregnancy at first insemination and number of inseminations per animal submitted for artificial insemination and potential predictor variables were investigated using a logistic regression model and a Poisson regression model, respectively. The results indicated that cows with severe claw lesions or an increasing somatic cell count after calving had a lower probability of pregnancy at first insemination and had a higher number of inseminations per animal submitted for artificial insemination than healthy cows. Variables representing a change in housing, production system, or milking system within the period from 6 mo before calving until establishment of a new pregnancy were significantly associated with decreased reproductive performance. No differences in fertility were observed between cows milked in an automatic milking system compared with cows milked conventionally. The results indicate that a change of system, rather than the actual type of milking or housing system negatively affects reproductive performance. Special attention should therefore be paid to the fertility of cows when the herd management is changing. It is also important to prevent claw lesions and increasing cell counts after calving to avoid a decrease in reproductive performance. PMID:23958022

  16. ?? versus ?? fate choice: counting the T-cell lineages at the branch point.

    PubMed

    Kreslavsky, Taras; Gleimer, Michael; Garbe, Annette I; von Boehmer, Harald

    2010-11-01

    Both ?? and ?? T cells develop in the thymus from a common progenitor. Historically distinguished by their T-cell receptor (TCR), these lineages are now defined on the basis of distinct molecular programs. Intriguingly, in many transgenic and knockout systems these programs are mismatched with the TCR type, leading to the development of ?? lineage cells driven by ??TCR and vice versa. These puzzling observations were recently explained by the demonstration that TCR signal strength, rather than TCR type per se, instructs lineage fate, with stronger TCR signal favoring ?? and weaker signal favoring ?? lineage fates. These studies also highlighted the ERK (extracellular signal regulated kinase)-Egr (early growth response)-Id3 (inhibitor of differentiation 3) axis as a potential molecular switch downstream of TCR that determines lineage choice. Indeed, removal of Id3 was sufficient to redirect TCR?? transgenic cells to the ?? lineage, even in the presence of strong TCR signal. However, in TCR non-transgenic Id3 knockout mice the overall number of ?? lineage cells was increased due to an outgrowth of a V?1V?6.3 subset, suggesting that not all ?? T cells depend on this molecular switch for lineage commitment. Thus, the ?? lineage may in fact be a collection of two or more lineages not sharing a common molecular program and thus equipollent to the ?? lineage. TCR signaling is not the only factor that is required for development of ?? and ?? lineage cells; other pathways, such as signaling from Notch and CXCR4 receptors, cooperate with the TCR in this process. PMID:20969592

  17. Racial/ethnic differences in CD4 T cell count and viral load at presentation for medical care and in follow-up after HIV-1 infection.

    PubMed

    Swindells, Susan; Cobos, Daniel G; Lee, Nancy; Lien, Elizabeth A; Fitzgerald, Ann P; Pauls, Jennifer S; Anderson, James R

    2002-09-01

    The baseline characteristics of antiretroviral-naive patients were compared by ethnic/racial groups. First CD4 T cell counts were lower for Latino (P = 0.0004) and black patients (P = 0.10) when compared with white patients. First HIV-1-RNA levels were higher in Latino patients (P = 0.08), who were also more likely to present with major opportunistic infections (P < 0.004). Once in care, changes in CD4 T cell counts and viral loads over time did not differ significantly between groups. PMID:12218399

  18. Osteogenic differentiation of amniotic fluid mesenchymal stromal cells and their bone regeneration potential

    PubMed Central

    Pipino, Caterina; Pandolfi, Assunta

    2015-01-01

    In orthopedics, tissue engineering approach using stem cells is a valid line of treatment for patients with bone defects. In this context, mesenchymal stromal cells of various origins have been extensively studied and continue to be a matter of debate. Although mesenchymal stromal cells from bone marrow are already clinically applied, recent evidence suggests that one may use mesenchymal stromal cells from extra-embryonic tissues, such as amniotic fluid, as an innovative and advantageous resource for bone regeneration. The use of cells from amniotic fluid does not raise ethical problems and provides a sufficient number of cells without invasive procedures. Furthermore, they do not develop into teratomas when transplanted, a consequence observed with pluripotent stem cells. In addition, their multipotent differentiation ability, low immunogenicity, and anti-inflammatory properties make them ideal candidates for bone regenerative medicine. We here present an overview of the features of amniotic fluid mesenchymal stromal cells and their potential in the osteogenic differentiation process. We have examined the papers actually available on this regard, with particular interest in the strategies applied to improve in vitro osteogenesis. Importantly, a detailed understanding of the behavior of amniotic fluid mesenchymal stromal cells and their osteogenic ability is desirable considering a feasible application in bone regenerative medicine. PMID:26029340

  19. A noninvasive approach to determine viscoelastic properties of an individual adherent cell under fluid flow.

    PubMed

    Qiu, Jun; Baik, Andrew D; Lu, X Lucas; Hillman, Elizabeth M C; Zhuang, Zhuo; Dong, Cheng; Guo, X Edward

    2014-04-11

    Mechanical properties of cells play an important role in their interaction with the extracellular matrix as well as the mechanotransduction process. Several in vitro techniques have been developed to determine the mechanical properties of cells, but none of them can measure the viscoelastic properties of an individual adherent cell in fluid flow non-invasively. In this study, techniques of fluid-structure interaction (FSI) finite element method and quasi-3-dimensional (quasi-3D) cell microscopy were innovatively applied to the frequently used flow chamber experiment, where an adherent cell was subjected to fluid flow. A new non-invasive approach, with cells at close to physiological conditions, was established to determine the viscoelastic properties of individual cells. The results showed an instantaneous modulus of osteocytes of 0.49 ± 0.11 kPa, an equilibrium modulus of 0.31 ± 0.044 kPa, and an apparent viscosity coefficient of 4.07 ± 1.23 kPas. This new quantitative approach not only provides an excellent means to measure cell mechanical properties, but also may help to elucidate the mechanotransduction mechanisms for a variety of cells under fluid flow stimulation. PMID:24581798

  20. Dual-Color Photon Counting Histogram Analysis of mRFP1 and EGFP in Living Cells

    PubMed Central

    Hillesheim, Lindsey N.; Chen, Yan; Müller, Joachim D.

    2006-01-01

    We investigate the potential of dual-color photon counting histogram (PCH) analysis to resolve fluorescent protein mixtures directly inside cells. Because of their small spectral overlap, we have chosen to look at the fluorescent proteins EGFP and mRFP1. We experimentally demonstrate that dual-color PCH quantitatively resolves a mixture of EGFP and mRFP1 in cells from a single measurement. To mimic the effect of protein association, we constructed a fusion protein of EGFP and mRFP1 (denoted EGFP-mRFP1). Fluorescence resonant energy transfer within the fusion protein alters the dual-channel brightness of the fluorophores. We describe a model for fluorescence resonant energy transfer effects on the brightness and incorporate it into dual-color PCH analysis. The model is verified using fluorescence lifetime measurements. Dual-color PCH analysis demonstrated that not all of the expressed EGFP-mRFP1 fusion proteins contained a fluorescent mRFP1 molecule. Fluorescence lifetime and emission spectra measurements confirmed this surprising result. Additional experiments show that the missing fluorescent fraction of mRFP1 is consistent with a dark state population of mRFP1. We successfully resolved this mixture of fusion proteins with a single dual-color PCH measurement. These results highlight the potential of dual-color PCH to directly detect and quantify protein mixtures in living cells. PMID:16980358

  1. Photoacoustic and photothermal detection of circulating tumor cells, bacteria and nanoparticles in cerebrospinal fluid in vivo and ex vivo

    PubMed Central

    Nedosekin, Dmitry A.; Juratli, Mazen A.; Sarimollaoglu, Mustafa; Moore, Christopher L.; Rusch, Nancy J.; Smeltzer, Mark S.; Zharov, Vladimir P.; Galanzha, Ekaterina I.

    2014-01-01

    Circulating cells, proteins, microparticles, and DNA in cerebrospinal fluid (CSF) are excellent biomarkers of many diseases, including cancer and infections. However, the sensitivity of existing methods is limited in their ability to detect rare CSF biomarkers at the treatable, early-stage of diseases. Here, we introduce novel CSF tests based on in vivo multicolor photoacoustic flow cytometry (PAFC) and ex vivo photothermal scanning cytometry. In the CSF of tumor-bearing mice, we molecularly detected in vivo circulating tumor cells (CTCs) before the development of breast cancer brain metastasis with 20-times higher sensitivity than with current assays. For the first time, we demonstrated assessing three pathways (i.e., blood, lymphatic, and CSF) of CTC dissemination, tracking nanoparticles in CSF in vivo and their imaging ex vivo. In label-free CSF samples, we counted leukocytes, erythrocytes, melanoma cells, and bacteria and imaged intracellular cytochromes, hemoglobin, melanin, and carotenoids, respectively. Taking into account the safety of PAFC, its translation for use in humans is expected to improve disease diagnosis beyond conventional detection limits. PMID:23681943

  2. Photoacoustic and photothermal detection of circulating tumor cells, bacteria and nanoparticles in cerebrospinal fluid in vivo and ex vivo.

    PubMed

    Nedosekin, Dmitry A; Juratli, Mazen A; Sarimollaoglu, Mustafa; Moore, Christopher L; Rusch, Nancy J; Smeltzer, Mark S; Zharov, Vladimir P; Galanzha, Ekaterina I

    2013-06-01

    Circulating cells, bacteria, proteins, microparticles, and DNA in cerebrospinal fluid (CSF) are excellent biomarkers of many diseases, including cancer and infections. However, the sensitivity of existing methods is limited in their ability to detect rare CSF biomarkers at the treatable, early-stage of diseases. Here, we introduce novel CSF tests based on in vivo photoacoustic flow cytometry (PAFC) and ex vivo photothermal scanning cytometry. In the CSF of tumor-bearing mice, we molecularly detected in vivo circulating tumor cells (CTCs) before the development of breast cancer brain metastasis with 20-times higher sensitivity than with current assays. For the first time, we demonstrated assessing three pathways (i.e., blood, lymphatic, and CSF) of CTC dissemination, tracking nanoparticles in CSF in vivo and their imaging ex vivo. In label-free CSF samples, we counted leukocytes, erythrocytes, melanoma cells, and bacteria and imaged intracellular cytochromes, hemoglobin, melanin, and carotenoids, respectively. Taking into account the safety of PAFC, its translation for use in humans is expected to improve disease diagnosis beyond conventional detection limits. PMID:23681943

  3. Flagellar Kinematics and Swimming of Algal Cells in Viscoelastic Fluids

    PubMed Central

    Qin, B.; Gopinath, A.; Yang, J.; Gollub, J. P.; Arratia, P. E.

    2015-01-01

    The motility of microorganisms is influenced greatly by their hydrodynamic interactions with the fluidic environment they inhabit. We show by direct experimental observation of the bi-flagellated alga Chlamydomonas reinhardtii that fluid elasticity and viscosity strongly influence the beating pattern - the gait - and thereby control the propulsion speed. The beating frequency and the wave speed characterizing the cyclical bending are both enhanced by fluid elasticity. Despite these enhancements, the net swimming speed of the alga is hindered for fluids that are sufficiently elastic. The origin of this complex response lies in the interplay between the elasticity-induced changes in the spatial and temporal aspects of the flagellar cycle and the buildup and subsequent relaxation of elastic stresses during the power and recovery strokes. PMID:25778677

  4. Effects of supplementation with a phytobiotics-rich herbal mixture on performance, udder health, and metabolic status of Holstein cows with various levels of milk somatic cell counts.

    PubMed

    Hashemzadeh-Cigari, F; Khorvash, M; Ghorbani, G R; Kadivar, M; Riasi, A; Zebeli, Q

    2014-12-01

    This study evaluated the effects of dietary supplementation of a novel phytobiotics-rich herbal mixture (PRHM) on feed intake, performance, udder health, ruminal fermentation, and plasma metabolites in cows with moderate or high somatic cell counts (SCC) in the milk. Twenty-four Holstein dairy cows (117 ± 26 d in milk and 46.3 ± 4.7 kg of milk/d at the start of the experiment) were blocked by parity and days in milk and split into 2 groups, based on SCC in the milk; 12 cows were with moderate SCC (260,000cells/mL), whereas 12 other cows had high levels of SCC (>500,000 cells/mL) in the milk. Within each SCC group, cows were blocked by milk yield and parity, and were randomly assigned to 2 different feeding regimens. Half of the cows in each SCC group (n=6) were supplemented with PRHM (185 g/cow per day, providing 12.4 g of phenolic compounds per day), and the other half (n=6) were not supplemented in their diets. The experiment lasted 36 d, whereby the first 24 d were used for adaptation to the diets and the last 12 d for sampling. Data showed that supplementation of PRHM decreased somatic cell score in the milk, indicating improved udder health of cows with high initial SCC, but not in cows with moderate SCC. Also, cows supplemented with PRHM consumed more feed DM, produced greater amounts of milk, and showed an improvement of feed utilization efficiency. However, these cows also lost more back-fat thickness during the experiment. Supplementation of PRHM increased fat- and energy-corrected milk yields in cows with high initial SCC, but not in cows with moderate SCC. Supplementation of PRHM decreased milk fat content, whereas other milk components were not affected by PRHM feeding. The PRHM supplementation decreased the acetate-to-propionate ratio in the rumen fluid, but increased ?-hydroxybutyrate and cholesterol concentration in the plasma, irrespective of the initial SCC level in the milk. Other plasma metabolites and liver enzymes were not affected by PRHM supplementation. Apparent nutrient digestibility did not differ among treatments. Overall, supplementation of PRHM seems to be an effective strategy to enhance performance and lower SCC, particularly in cows having high SCC levels in the milk. Further research is warranted to evaluate long-term effects of PRHM supplementation, especially with regard to metabolic health status and reproduction. PMID:25306268

  5. Transcript counting in single cells reveals dynamics of rDNA transcription

    Microsoft Academic Search

    Rui Zhen Tan; Alexander van Oudenaarden

    2010-01-01

    Most eukaryotes contain many tandem repeats of ribosomal RNA genes of which only a subset is transcribed at any given time. Current biochemical methods allow for the determination of the fraction of transcribing repeats (ON) versus non-transcribing repeats (OFF) but do not provide any dynamical information and obscure any transcription activity at the single-cell level. Here, we use a fluorescence

  6. Electrokinetic Focusing and Separation of Mammalian Cells in Conductive Biological Fluids

    PubMed Central

    Gao, Jian Gao; Riahi, Reza; Sin, Mandy L. Y.; Zhang, Shufeng; Wong, Pak Kin

    2014-01-01

    Active manipulation of cells, such as trapping, focusing, and isolation, is essential for various bioanalytical applications. Herein, we report a hybrid electrokinetic technique for manipulating mammalian cells in physiological fluids. This technique applies a combination of negative dielectrophoretic force and hydrodynamic drag force induced by electrohydrodynamics, which is effective in conductive biological fluids. With a three-electrode configuration, the stable equilibrium positions of cells can be adjusted for separation and focusing applications. Cancer cells and white blood cells can be positioned and isolated into specific locations in the microchannel under both static and dynamic flow conditions. To investigate the sensitivity of the hybrid electrokinetic process, AC voltage, frequency, and bias dependences of the cell velocity were studied systematically. The applicability of the hybrid electrokinetic technique for manipulating cells in physiological samples is demonstrated by continuous focusing human breast adenocarcinoma spiked in urine, buffy coats, and processed blood samples with 98% capture efficiency. PMID:22937529

  7. Sex and species differences in plasma testosterone and in counts of androgen receptor-positive cells in key brain regions of Sceloporus lizard species that differ in aggression

    PubMed Central

    Hews, Diana K.; Hara, Erina; Anderson, Maurice C.

    2012-01-01

    We studied neuroendocrine correlates of aggression differences in adults of two Sceloporus lizard species. These species differ in the degree of sex difference in aggressive color signals (belly patches) and in aggression: S. undulatus (males blue, high aggression; females white, low aggression) and S. virgatus (both sexes white, lower aggression). We measured plasma testosterone and counted cells expressing androgen receptor-like immunoreactivity to the affinity-purified polyclonal AR antibody, PG-21, in three brain regions of breeding season adults. Male S. undulatus had the highest mean plasma testosterone and differed significantly from conspecific females. In contrast, there was no sex difference in plasma testosterone concentrations in S. virgatus. Male S. undulatus also had the highest mean number of AR-positive cells in the preoptic area: the sexes differed in S. undulatus but not in S. virgatus, and females of the two species did not differ. In the ventral medial hypothalamus, S. undulatus males had higher mean AR cell counts compared to females, but again there was no sex difference in S. virgatus. In the habenula, a control brain region, the sexes did not differ, and although the sex by species interaction significant was not significant, there was a trend (p = 0.050) for S. virgatus to have higher mean AR cell counts than S. undulatus. Thus hypothalamic AR cell counts paralleled sex and species differences in aggression, as did mean plasma testosterone levels in these breeding-season animals. PMID:22230767

  8. Computational Fluid Dynamic Simulation of Aggregation of Deformable Cells in a Shear Flow

    Microsoft Academic Search

    Prosenjit Bagchi; Paul C. Johnson; Aleksander S. Popel

    2005-01-01

    We present computational fluid dynamic (CFD) simulation of aggregation of two deform- able cells in a shear flow. This work is motivated by an attempt to develop computational models of aggregation of red blood cells (RBCs). Aggregation of RBCs is a major deter- minant of blood viscosity in microcirculation under physiological and pathological con- ditions. Deformability of the RBCs plays

  9. Characterization of Peripheral Blood and Peritoneal Fluid Mononuclear Cell Subsets in Fertile and Infertile Women

    Microsoft Academic Search

    Michael S. Opsahl; Clifford C. Hayslip; Thomas A. Klein; Dean S. Cunningham

    1994-01-01

    Mononuclear cell subpopulations from the peripheral blood (PB) and peritoneal fluid (PF) of fertile and infertile women were quantified by flow cytometry using a double-staining monoclonal antibody technique. No differences in the percentage distribution of mononuclear cells between fertile and infertile women were demonstrated when either the PB constituents or the PF components were compared to one another. When the

  10. Intracellular [Na +], Na + pathways, and fluid transport in cultured bovine corneal endothelial cells

    Microsoft Academic Search

    Kunyan Kuang; Yansui Li; Maimaiti Yiming; José M. Sánchez; Pavel Iserovich; E. J. Cragoe; Friedrich P. J. Diecke; Jorge Fischbarg

    2004-01-01

    The mechanism of fluid transport across corneal endothelium remains unclear. We examine here the relative contributions of cellular mechanisms of Na+ transport and the homeostasis of intracellular [Na+] in cultured bovine corneal endothelial cells, and the influence of ambient Na+ and HCO3?on the deturgescence of rabbit cornea. Bovine corneal endothelial cells plated on glass coverslips were incubated for 60 min

  11. Regulation of bronchoalveolar lavage fluids cell function by the immunomodulatory agents from Cordyceps sinensis

    Microsoft Academic Search

    Yuh-Chi Kuo; Wei-Jern Tsai; Jir-Yenn Wang; Shi-Chung Chang; Ching-Yuang Lin; Ming-Shi Shiao

    2001-01-01

    Cordyceps sinensis (C. sinensis) is one of the well known fungi used in traditional Chinese medicine for treatment asthma and bronchial and lung inflammation. In this study, effects of C. sinensis methanolic extracts on bronchoalveolar lavage fluids (BALF) cells proliferation, inflammatory cytokines production, and genes expression were evaluated. The proliferative response of BALF cells to lipopolysaccharide (LPS) was determined by

  12. Counting Coins

    NSDL National Science Digital Library

    thorsen

    2012-11-24

    We are learning about money and how to count coins. We need to learn about coins so we can pay for things we need to buy. These activities will help you practice counting money. Remember to record your learning as you work! Coin Paper We have been learning about coins. Listen to the coin song to remember the names of U.S. coins. U.S. Coin Song Before we can count coins, we need to know the names of the different coins and how much each coin is worth. Click the link below to review ...

  13. Prognostic significance of bcl-xL gene expression and apoptotic cell counts in follicular lymphoma

    Microsoft Academic Search

    Wei-Li Zhao; Marjan Ertault Daneshpouy; Nicolas Mounier; Josette Briere; Christophe Leboeuf; Louis-Francois Plassa; Elisabeth Turpin; Jean-Michel Cayuela; Jean-Claude Ameisen; Christian Gisselbrecht; Anne Janin

    2003-01-01

    bcl-xL, a member of the Bcl-2 family, exerts an antiapoptotic effect on lympho- cytes. To assess its clinical significance in patients with follicular lymphoma, real- time quantitative reverse transcription- polymerase chain reaction (RT-PCR) anal- ysis of bcl-xL gene expression was investigated in whole lymph node sec- tions and laser-microdissected lymphoma cells of 27 patients. Compared with 10 patients with reactive

  14. Fluid biopsy for circulating tumor cell identification in patients with early-and late-stage non-small cell lung cancer: a glimpse into lung cancer biology

    NASA Astrophysics Data System (ADS)

    Wendel, Marco; Bazhenova, Lyudmila; Boshuizen, Rogier; Kolatkar, Anand; Honnatti, Meghana; Cho, Edward H.; Marrinucci, Dena; Sandhu, Ajay; Perricone, Anthony; Thistlethwaite, Patricia; Bethel, Kelly; Nieva, Jorge; van den Heuvel, Michel; Kuhn, Peter

    2012-02-01

    Circulating tumor cell (CTC) counts are an established prognostic marker in metastatic prostate, breast and colorectal cancer, and recent data suggest a similar role in late stage non-small cell lung cancer (NSCLC). However, due to sensitivity constraints in current enrichment-based CTC detection technologies, there are few published data about CTC prevalence rates and morphologic heterogeneity in early-stage NSCLC, or the correlation of CTCs with disease progression and their usability for clinical staging. We investigated CTC counts, morphology and aggregation in early stage, locally advanced and metastatic NSCLC patients by using a fluid-phase biopsy approach that identifies CTCs without relying on surface-receptor-based enrichment and presents them in sufficiently high definition (HD) to satisfy diagnostic pathology image quality requirements. HD-CTCs were analyzed in blood samples from 78 chemotherapy-naïve NSCLC patients. 73% of the total population had a positive HD-CTC count (>0 CTC in 1 mL of blood) with a median of 4.4 HD-CTCs mL-1 (range 0-515.6) and a mean of 44.7 (±95.2) HD-CTCs mL-1. No significant difference in the medians of HD-CTC counts was detected between stage IV (n = 31, range 0-178.2), stage III (n = 34, range 0-515.6) and stages I/II (n = 13, range 0-442.3). Furthermore, HD-CTCs exhibited a uniformity in terms of molecular and physical characteristics such as fluorescent cytokeratin intensity, nuclear size, frequency of apoptosis and aggregate formation across the spectrum of staging. Our results demonstrate that despite stringent morphologic inclusion criteria for the definition of HD-CTCs, the HD-CTC assay shows high sensitivity in the detection and characterization of both early- and late-stage lung cancer CTCs. Extensive studies are warranted to investigate the prognostic value of CTC profiling in early-stage lung cancer. This finding has implications for the design of extensive studies examining screening, therapy and surveillance in lung cancer patients.

  15. Genetic variants associated with the white blood cell count in 13,923 subjects in the eMERGE Network

    PubMed Central

    McDavid, Andrew; Weston, Noah; Nelson, Sarah C.; Zheng, Xiuwen; Hart, Eugene; de Andrade, Mariza; Kullo, Iftikhar J.; McCarty, Catherine A.; Doheny, Kimberly F.; Pugh, Elizabeth; Kho, Abel; Hayes, M. Geoffrey; Pretel, Stephanie; Saip, Alexander; Ritchie, Marylyn D.; Crawford, Dana C.; Crane, Paul K.; Newton, Katherine; Li, Rongling; Mirel, Daniel B.; Crenshaw, Andrew; Larson, Eric B.; Carlson, Chris S.; Jarvik, Gail P.

    2013-01-01

    White blood cell count (WBC) is unique among identified inflammatory predictors of chronic disease in that it is routinely measured in asymptomatic patients in the course of routine patient care. We led a genome-wide association analysis to identify variants associated with WBC levels in 13,923 subjects in the electronic Medical Records and Genomics (eMERGE) Network. We identified two regions of interest that were each unique to subjects of genetically determined ancestry to the African continent (AA) or to the European continent (EA). WBC varies among different ancestry groups. Despite being ancestry specific, these regions were identifiable in the combined analysis. In AA subjects, the region surrounding the Duffy antigen/chemokine receptor gene (DARC) on 1q21 exhibited significant association (p value = 6.71e–55). These results validate the previously reported association between WBC and of the regulatory variant rs2814778 in the promoter region, which causes the Duffy negative phenotype (Fy?/?). A second missense variant (rs12075) is responsible for the two principal antigens, Fya and Fyb of the Duffy blood group system. The two variants, consisting of four alleles, act in concert to produce five antigens and subsequent phenotypes. We were able to identify the marginal and novel interaction effects of these two variants on WBC. In the EA subjects, we identified significantly associated SNPs tagging three separate genes in the 17q21 region: (1) GSDMA, (2) MED24, and (3) PSMD3. Variants in this region have been reported to be associated with WBC, neutrophil count, and inflammatory diseases including asthma and Crohn’s disease. PMID:22037903

  16. Impact of red blood cells count and high density lipoproteins with the prevalence and extent of coronary artery disease.

    PubMed

    Schaffer, Alon; Verdoia, Monica; Cassetti, Ettore; Barbieri, Lucia; Perrone-Filardi, Pasquale; Marino, Paolo; De Luca, Giuseppe

    2015-07-01

    We have hypothesized that high red blood cells (RBC) count can potentially play an atheroprotective role in patients with coronary atherosclerosis. We, therefore, have investigated the relationship between high density lipoproteins cholesterol (HDL-C) and RBC levels in patients undergoing coronary angiography. Coronary artery disease (CAD) is a major cause of mortality. Impaired lipid profile represents a major risk factor for atherosclerosis. High density lipoprotein (HDL) is a key factor in atherosclerosis disease development. RBC can mimic HDL's reverse cholesterol transportation with a potential atheroprotective role. Coronary angiography has been evaluated in 3,534 patients. Fasting samples were collected for haematology and lipids levels assessment. Coronary disease was defined for at least 1 vessel stenosis >50 %. Patients were divided according to HDL-C and RBC tertiles. Lower HDL-C was significantly associated to the prevalence of CAD (84.8 vs 78.5 vs 67.3 %, p ? 0.001; adjusted OR [95 % CI] = 1.55 [1.3-1.8], p < 0.001) and severe CAD (30 % vs 30 % vs 24.4 %, p = 0.002; adjusted OR [95 % CI] = 1.08 [1.01-1.16], p = 0.02), this relationship was maintained even dividing our population according to RBC tertiles (p < 0.001).In conclusion, HDL-C levels are directly related to RBC count and inversely to the prevalence and extent of coronary disease. Higher RBC levels can reduce the risk of CAD in patients with lower HDL-C levels, suggesting an important atheroprotective role. PMID:25680891

  17. Clock Counting

    NSDL National Science Digital Library

    Ms. McDuffee

    2008-11-12

    Students will practice telling time. Review clock counting with the interactive clock. Now match the clocks. Move over the hour clock to see if you chose correctly. Click the arrows to match the dragon clock to the written time. ...

  18. High-pressure cell for neutron reflectometry of supercritical and subcritical fluids at solid interfaces

    SciTech Connect

    Carmichael, Justin R [ORNL; Rother, Gernot [ORNL; Browning, Jim [ORNL; Ankner, John Francis [ORNL; Banuelos, Jose Leo [ORNL; Anovitz, Lawrence {Larry} M [ORNL; Wesolowski, David J [ORNL; Cole, David [Ohio State University

    2012-01-01

    A new high-pressure cell design for use in neutron reflectometry (NR) for pressures up to 50 MPa and a temperature range of 300 473 K is described. The cell design guides the neutron beam through the working crystal without passing through additional windows or the bulk fluid, which provides for a high neutron transmission, low scattering background, and low beam distortion. The o-ring seal is suitable for a wide range of subcritical and supercritical fluids and ensures high chemical and pressure stability. Wafers with a diameter of 5.08 cm (2 in.) and 5 mm or 10 mm thickness can be used with the cells, depending on the required pressure and momentum transfer range. The fluid volume in the sample cell is very small at about 0.1 ml, which minimizes scattering background and stored energy. The cell design and pressure setup for measurements with supercritical fluids are described. NR data are shown for silicon/silicon oxide and quartz wafers measured against air and subsequently within the high-pressure cell to demonstrate the neutron characteristics of the high-pressure cell. Neutron reflectivity data for supercritical CO2 in contact with quartz and Si/SiO2 wafers are also shown.

  19. Automated Static Culture System Cell Module Mixing Protocol and Computational Fluid Dynamics Analysis

    NASA Technical Reports Server (NTRS)

    Kleis, Stanley J.; Truong, Tuan; Goodwin, Thomas J,

    2004-01-01

    This report is a documentation of a fluid dynamic analysis of the proposed Automated Static Culture System (ASCS) cell module mixing protocol. The report consists of a review of some basic fluid dynamics principles appropriate for the mixing of a patch of high oxygen content media into the surrounding media which is initially depleted of oxygen, followed by a computational fluid dynamics (CFD) study of this process for the proposed protocol over a range of the governing parameters. The time histories of oxygen concentration distributions and mechanical shear levels generated are used to characterize the mixing process for different parameter values.

  20. Accuracy of Methods Using Somatic Cell Count and N-Acetyl-?-D-Glucosaminidase Activity in Milk to Assess the Bacteriological Cure of Bovine Clinical Mastitis

    Microsoft Academic Search

    S. PYORALAand E. PYORALA; E. Pyörälä

    1997-01-01

    This study examined the capability of milk somatic cell count (SCC) andNAGase activity to discriminate between quarters that had been cured versus those that had not been cured at 4 wk after antimicrobial therapy for clinical mastitis. The distribution of microorganisms that were isolated before therapy from 630 quarters with mastitis was as follows: 225 strains of Staphylococcus aureus, 96

  1. Genome-Wide Association Study of White Blood Cell Count in 16,388 African Americans: the Continental Origins and Genetic Epidemiology Network (COGENT)

    Microsoft Academic Search

    Alexander P. Reiner; Guillaume Lettre; Michael A. Nalls; Santhi K. Ganesh; Rasika Mathias; Melissa A. Austin; Eric Dean; Sampath Arepalli; Angela Britton; Zhao Chen; David Couper; J. David Curb; Charles B. Eaton; Myriam Fornage; Struan F. A. Grant; Tamara B. Harris; Dena Hernandez; Naoyuki Kamatini; Brendan J. Keating; Michiaki Kubo; Andrea LaCroix; Leslie A. Lange; Simin Liu; Kurt Lohman; Yan Meng; Emile R. Mohler; Solomon Musani; Yusuke Nakamura; Christopher J. ODonnell; Yukinori Okada; Cameron D. Palmer; George J. Papanicolaou; Kushang V. Patel; Andrew B. Singleton; Atsushi Takahashi; Hua Tang; Herman A. Taylor; Kent Taylor; Cynthia Thomson; Lisa R. Yanek; Lingyao Yang; Elad Ziv; Alan B. Zonderman; Aaron R. Folsom; Michele K. Evans; Yongmei Liu; Diane M. Becker; Beverly M. Snively; James G. Wilson

    2011-01-01

    Total white blood cell (WBC) and neutrophil counts are lower among individuals of African descent due to the common African-derived “null” variant of the Duffy Antigen Receptor for Chemokines (DARC) gene. Additional common genetic polymorphisms were recently associated with total WBC and WBC sub-type levels in European and Japanese populations. No additional loci that account for WBC variability have been

  2. Increases in platelet and red cell counts, blood viscosity, and arterial pressure during mild surface cooling: factors in mortality from coronary and cerebral thrombosis in winter

    Microsoft Academic Search

    W R Keatinge; S R Coleshaw; F Cotter; M Mattock; M Murphy; R Chelliah

    1984-01-01

    Six hours of mild surface cooling in moving air at 24 degrees C with little fall in core temperature (0.4 degree C) increased the packed cell volume by 7% and increased the platelet count and usually the mean platelet volume to produce a 15% increase in the fraction of plasma volume occupied by platelets. Little of these increases occurred in

  3. Genetic and Phenotypic Correlations Between Milk Coagulation Properties, Milk Production Traits, Somatic Cell Count, Casein Content, and pH of Milk

    Microsoft Academic Search

    T. Ikonen; S. Morri; A.-M. Tyrisevä; O. Ruottinen; M. Ojala

    2004-01-01

    Genetic and phenotypic correlations between milk coagulation properties (MCP: coagulation time and curd firmness), milk yield, fat content, protein content, ln(somatic cell count) (SCS), casein content, and pH of milk and heritability of these traits were estimated from data consisting of milk samples of 4664 Finnish Ayrshire cows sired by 91 bulls. In addition, differences in average estimated breeding values

  4. APOE Polymorphism Is Associated with C-reactive Protein Levels but Not with White Blood Cell Count: Dong-gu Study and Namwon Study.

    PubMed

    Yun, Yong-Woon; Kweon, Sun-Seog; Choi, Jin-Su; Rhee, Jung-Ae; Lee, Young-Hoon; Nam, Hae-Sung; Jeong, Seul-Ki; Park, Kyeong-Soo; Ryu, So-Yeon; Choi, Seong-Woo; Kim, Hee Nam; Cauley, Jane A; Shin, Min-Ho

    2015-07-01

    We evaluated the association of the APOE polymorphism with serum C-reactive protein levels and white blood cell count in two large population-based studies in Korean. The datasets included the Dong-gu study (n = 8,893) and the Namwon Study (n = 10,032). APOE genotypes were identified by polymerase chain reaction-restriction fragment length polymorphism. Multivariable linear regression analysis was performed to evaluate the relationship of APOE genotypes with C-reactive protein levels and white blood cell count with adjustments for age, sex, body mass index, smoking, diabetes, hypertension, and serum lipids. In the multivariate model, carriers of E3E4 or E4E4 genotype had significantly lower C-reactive protein levels compared with carriers of E3E3 genotype group (0.50 mg/L vs. 0.67 mg/L; 0.37 mg/L vs. 0.67 mg/L, respectively, for the Dong-gu Study and 0.47 mg/L vs. 0.66 mg/L; 0.45 mg/L vs. 0.66 mg/L, respectively, for the Namwon Study). However, there was no difference in white blood cell count among APOE genotypes. We found that the APOE E4 allele is associated with lower C-reactive protein levels, but not white blood cell count. Our results suggest that APOE genotype may influence C-reactive protein levels through non-inflammatory pathway. PMID:26130946

  5. Ca2+ Signaling and Regulation of Fluid Secretion in Salivary Gland Acinar Cells

    PubMed Central

    Ambudkar, Indu S.

    2014-01-01

    Neurotransmitter stimulation of plasma membrane receptors stimulates salivary gland fluid secretion via a complex process that is determined by coordinated temporal and spatial regulation of several Ca2+ signaling processes as well as ion flux systems. Studies over the past four decades have demonstrated that Ca2+ is a critical factor in the control of salivary gland function. Importantly, critical components of this process have now been identified, including plasma membrane receptors, calcium channels, and regulatory proteins. The key event in activation of fluid secretion is an increase in intracellular [Ca2+] ([Ca2+]i) triggered by IP3-induced the release of Ca2+ from ER via the IP3R. This increase regulates the ion fluxes required to drive vectorial fluid secretion. IP3Rs determine the site of initiation and the pattern of [Ca2+]i signal in the cell. However, Ca2+ entry into the cell is required to sustain the elevation of [Ca2+]i and fluid secretion. This Ca2+ influx pathway, store-operated calcium influx pathway (SOCE), has been studied in great detail and the regulatory mechanisms as well as key molecular components have now been identified. Orai1, TRPC1, and STIM1 are critical components of SOCE and among these, Ca2+ entry via TRPC1 is a major determinant of fluid secretion. The receptor-evoked Ca2+ signal in salivary gland acinar cells is unique in that it starts at the apical pole and then rapidly increases across the cell. The basis for the polarized Ca2+ signal can be ascribed to the polarized arrangement of the Ca2+ channels, transporters, and signaling proteins. Distinct localization of these proteins in the cell suggests compartmentalization of Ca2+ signals during regulation of fluid secretion. This chapter will discuss new concepts and findings regarding the polarization and control of Ca2+ signals in the regulation of fluid secretion. PMID:24646566

  6. Emergent long-range couplings in arrays of fluid cells

    SciTech Connect

    Abraham, Douglas Bruce [Los Alamos National Lab. (LANL), Los Alamos, NM (United States)

    2014-08-07

    We present a system exhibiting extraordinarily long-range cooperative effects, on a length scale far exceeding the bulk correlation length. We give a theoretical explanation of these phenomena based on the mesoscopic picture of phase coexistence in finite systems, which is confirmedly Monte Carlo (MC) simulation studies. Our work demonstrates that such action-at-a-distance can occur in classical systems involving simple or complex fluids, such as colloid-polymer mixtures, or ferromagnets.

  7. Trends in and correlates of CD4+ cell count at antiretroviral therapy initiation after changes in national ART guidelines in Rwanda

    PubMed Central

    Mutimura, Eugene; Addison, Diane; Anastos, Kathryn; Hoover, Donald; Dusingize, Jean Claude; Karenzie, Ben; Izimukwiye, Isabelle; Mutesa, Leo; Nsanzimana, Sabin; Nashi, Denis

    2015-01-01

    Background Initiation of antiretroviral therapy (ART) in the advanced stages of HIV infection remains a major challenge in sub-Saharan Africa. This study was conducted to better understand barriers and enablers to timely ART initiation in Rwanda where ART coverage is high and national ART eligibility guidelines first expanded in 2007–2008. Methods Using data on 6326 patients (?15 years) at five Rwandan clinics, we assessed trends and correlates of CD4+ cell count at ART initiation and the proportion initiating ART with advanced HIV disease (CD4+ <200 cells/µl or WHO stage IV). Results Out of 6326 patients, 4486 enrolling in HIV care initiated ART with median CD4+ cell count of 211 cells/µl [interquartile range: 131–300]. Median CD4+ cell counts at ART initiation increased from 183 cells/µl in 2007 to 293 cells/µl in 2011–2012, and the proportion with advanced HIV disease decreased from 66.2 to 29.4%. Factors associated with a higher odds of advanced HIV disease at ART initiation were male sex [adjusted odds ratios (AOR) = 1.7; 95% confidence interval (CI): 1.3–2.1] and older age (AOR46–55+ vs. <25 = 2.3; 95% CI: 1.2–4.3). Among those initiating ART more than 1 year after enrollment in care, those who had a gap in care of 12 or more months prior to ART initiation had higher odds of advanced HIV disease (AOR = 5.2; 95% CI: 1.2–21.1). Conclusion Marked improvements in the median CD4+ cell count at ART initiation and proportion initiating ART with advanced HIV disease were observed following the expansion of ART eligibility criteria in Rwanda. However, sex disparities in late treatment initiation persisted through 2011–2012, and appeared to be driven by later diagnosis and/or delayed linkage to care among men. PMID:25562492

  8. Short-Chain PEG Mixed-Monolayer Protected Gold Clusters Increase Clearance and Red Blood Cell Counts

    PubMed Central

    Simpson, Carrie A.; Agrawal, Amanda C.; Balinski, Andrzej; Harkness, Kellen M.; Cliffel, David E.

    2011-01-01

    Monolayer-protected gold nanoparticles have great potential as novel building blocks for the design of new drugs and therapeutics based on the easy ability to multifunctionalize them for biological targeting and drug activity. In order to create nanoparticles that are biocompatible in vivo, poly-ethylene glycol functional groups have been added to many previous multifunctionalized particles to eliminate non-specific binding. Recently, monolayer-protected gold nanoparticles with mercaptoglycine functionalities were shown to elicit deleterious effects on the kidney in vivo that were eliminated by incorporating a long-chain, mercapto-undecyl-tetraethylene glycol, at very high loadings into a mixed monolayer. These long-chain PEGs induced an immune response to the particle presumably generating an anti-PEG antibody as seen in other long-chain PEG-ylated nanoparticles in vivo. In the present work, we explore the in vivo effects of high and low percent ratios of a shorter chain, mercapto-tetraethylene glycol, within the monolayer using simple place-exchange reactions. The shorter chain PEG MPCs were expected to have better water solubility due to elimination of the alkyl chain, no toxicity, and long-term circulation in vivo. Shorter chain lengths at lower concentrations should not trigger the immune system into creating an anti-PEG antibody. We found that a 10% molar exchange of this short chain PEG within the monolayer met three of the desired goals: high water solubility, no toxicity, and no immune response as measured by white blood cell counts, but none of the short chain PEG mixed monolayer compositions enabled the nanoparticles to have a long circulation time within the blood as compared to mercapto-undecyl-ethylene glycol, which had a residence time of 4 weeks. We also compared the effects of a hydroxyl versus a carboxylic acid terminal functional group on the end of the PEG thiol on both clearance and immune response. The results indicate that short-chain length PEGs, regardless of termini, increase clearance rates compared to the previous long-chain PEG studies while carboxylated-termini increase red blood cell counts at high loadings. Given these findings, short-chain, alcohol-terminated PEG, exchanged at 10% was identified as a potential nanoparticle for further in vivo applications requiring short circulation lifetimes with desired features of no toxicity, no immune response, and high water solubility. PMID:21473648

  9. Fluid intake and incidence of renal cell carcinoma in UK women

    PubMed Central

    Allen, N E; Balkwill, A; Beral, V; Green, J; Reeves, G

    2011-01-01

    Background: It has been suggested that the apparent protective effect of alcohol intake on renal cell carcinoma may be due to the diluting effect of carcinogens by a high total fluid intake. We assessed the association between intakes of total fluids and of specific beverages on the risk of renal cell carcinoma in a large prospective cohort of UK women. Methods: Information on beverage consumption was obtained from a questionnaire sent ?3 years after recruitment into the Million Women Study. Cox proportional hazards models were used to estimate relative risks (RRs) and 95% confidence intervals (CIs) for renal cell carcinoma associated with beverage consumption adjusted for age, region of residence, socioeconomic status, smoking, and body mass index. Results: After an average of 5.2 years of follow-up, 588 cases of renal cell carcinoma were identified among 779?369 women. While alcohol intake was associated with a reduced risk of renal cell carcinoma (RR for ?2 vs <1 drink per day: 0.76; 95% CI: 0.61–0.96; P for trend=0.02), there was no association with total fluid intake (RR for ?12 vs <7 drinks per day: 1.15; 95% CI: 0.91–1.45; P for trend=0.3) or with intakes of specific beverages. Conclusions: The apparent protective effect of alcohol on the risk of renal cell carcinoma is unlikely to be related to a high fluid intake. PMID:21407222

  10. Characteristics of Human Amniotic Fluid Mesenchymal Stem Cells and Their Tropism to Human Ovarian Cancer

    PubMed Central

    Li, Liru; Wang, Dejun; Zhou, Jun; Cheng, Yan; Liang, Tian; Zhang, Guangmei

    2015-01-01

    The mesenchymal stem cells (MSCs) derived from amniotic fluid (AF) have become an attractive stem cells source for cell-based therapy because they can be harvested at low cost and avoid ethical disputes. In human research, stem cells derived from AF gradually became a hot research direction for disease treatment, specifically for their plasticity, their reduced immunogenicity and their tumor tropism regardless of the tumor size, location and source. Our work aimed to obtain and characterize human amniotic fluid mesenchymal stem cells (AFMSCs) and detect their ovarian cancer tropsim in nude mice model. Ten milliliters of twenty independent amniotic fluid samples were collected from 16-20 week pregnant women who underwent amniocentesis for fetal genetic determination in routine prenatal diagnosis in the first affiliated hospital of Harbin medical university. We successfully isolated the AFMSCs from thirteen of twenty amniotic fluid samples. AFMSCs presented a fibroblastic-like morphology during the culture. Flow cytometry analyses showed that the cells were positive for specific stem cell markers CD73,CD90, CD105, CD166 and HLA-ABC (MHC class I), but negative for CD 45,CD40, CD34, CD14 and HLA-DR (MHC class II). RT-PCR results showed that the AFMSCs expressed stem cell marker OCT4. AFMSCs could differentiate into bone cells, fat cells and chondrocytes under certain conditions. AFMSCs had the high motility to migrate to ovarian cancer site but didn’t have the tumorigenicity. This study enhances the possibility of AFMSCs as drug carrier in human cell-based therapy. Meanwhile, the research emphasis in the future can also put in targeting therapy of ovarian cancer. PMID:25880317

  11. Clinical Outcome of HIV Viraemic Controllers and Noncontrollers with Normal CD4 Counts Is Exclusively Determined by Antigen-Specific CD8+ T-Cell-Mediated HIV Suppression

    PubMed Central

    Tansiri, Yada; Rowland-Jones, Sarah L.; Ananworanich, Jintanat; Hansasuta, Pokrath

    2015-01-01

    In this cross-sectional study we evaluated T-cell responses using several assays to determine immune correlates of HIV control that distinguish untreated viraemic controllers (VC) from noncontrollers (NC) with similar CD4 counts. Samples were taken from 65 ART-naïve chronically HIV-infected VC and NC from Thailand with matching CD4 counts in the normal range (>450 cells/?l). We determined HIVp24-specific T-cell responses using standard Interferon-gamma (IFN?) ELISpot assays, and compared the functional quality of HIVp24-specific CD8+ T-cell responses using polychromatic flow cytometry. Finally, in vitro HIV suppression assays were performed to evaluate directly the activity of CD8+ T cells in HIV control. Autologous CD4+ T cells were infected with primary patient-derived HIV isolates and the HIV suppressive activity of CD8+ T cells was determined after co-culture, measuring production of HIVp24 Ag by ELISA. The HIVp24-specific T-cell responses of VC and NC could not completely be differentiated through measurement of IFN?-producing cells using ELISpot assays, nor by the absolute cell numbers of polyfunctional HIVp24-specific CD8+ T cells. However, in vitro HIV suppression assays showed clear differences between VC and NC. HIV suppressive activity, mediated by either ex vivo unstimulated CD8+ T cells or HIVp24-specific T-cell lines, was significantly greater using cells from VC than NC cells. Additionally, we were able to demonstrate a significant correlation between the level of HIV suppressive activity mediated by ex vivo unstimulated CD8+ T cells and plasma viral load (pVL) (Spearman r = -0.7345, p = 0.0003). This study provides evidence that in vitro HIV suppression assays are the most informative in the functional evaluation of CD8+ T-cell responses and can distinguish between VC and NC. PMID:25764310

  12. Compressed Counting

    E-print Network

    Li, Ping

    2008-01-01

    Counting is among the most fundamental operations in computing. For example, counting the pth frequency moment has been a very active area of research, in theoretical computer science, databases, and data mining. When p=1, the task (i.e., counting the sum) can be accomplished using a simple counter. Compressed Counting (CC) is proposed for efficiently computing the pth frequency moment of a data stream signal A_t, where 0= 0, which includes the strict Turnstile model as a special case. For natural data streams encountered in practice, this restriction is minor. The underly technique for CC is what we call skewed stable random projections, which captures the intuition that, when p=1 a simple counter suffices, and when p = 1+/\\Delta with small \\Delta, the sample complexity of a counter system should be low (continuously as a function of \\Delta). We show at small \\Delta the sample complexity (number of projections) k = O(1/\\epsilon) instead of O(1/\\epsilon^2). Compressed Counting can serve a basic building block...

  13. Detection of metallothionein in bronchoalveolar cells and lavage fluid following repeated cadmium inhalation

    SciTech Connect

    Hart, B.A.; Garvey, J.S.

    1986-08-01

    Metallothionein (MT) levels were measured by radioimmunoassay in bronchoalveolar cells and fluids derived from rat lungs following 0, 8, 12, 18, or 24 exposures to Cd acetate aerosols. Alveolar cells methallothionein levels increased from a base line value of 0.35 ..mu..g MT/mg protein in control rats to a value of 21.73 ..mu..g MT/mg protein in rats exposed 24 times to Cd aerosols. Cell-free lavage fluid levels of methallothionein also increased in response to frequency of Cd exposure both but 5 to 10-fold lower than in the free alveolar cells. Gel chromatographic analysis of free alveolar cells from Cd-exposed animals revealed the presence of a 11,600-Da Cd-binding protein with an absorption maximum of 254 nm. Metallothionein analyses of purified populations of alveolar macrophages and polymorphonuclear leukocytes from exposed animals demonstrated that macrophages but not polymorphonuclear leukocytes contained metallothionein.

  14. Normal Distribution of CD8+ T-Cell-Derived ELISPOT Counts within Replicates Justifies the Reliance on Parametric Statistics for Identifying Positive Responses

    PubMed Central

    Karulin, Alexey Y.; Caspell, Richard; Dittrich, Marcus; Lehmann, Paul V.

    2015-01-01

    Accurate assessment of positive ELISPOT responses for low frequencies of antigen-specific T-cells is controversial. In particular, it is still unknown whether ELISPOT counts within replicate wells follow a theoretical distribution function, and thus whether high power parametric statistics can be used to discriminate between positive and negative wells. We studied experimental distributions of spot counts for up to 120 replicate wells of IFN-? production by CD8+ T-cell responding to EBV LMP2A (426 – 434) peptide in human PBMC. The cells were tested in serial dilutions covering a wide range of average spot counts per condition, from just a few to hundreds of spots per well. Statistical analysis of the data using diagnostic Q-Q plots and the Shapiro-Wilk normality test showed that in the entire dynamic range of ELISPOT spot counts within replicate wells followed a normal distribution. This result implies that the Student t-Test and ANOVA are suited to identify positive responses. We also show experimentally that borderline responses can be reliably detected by involving more replicate wells, plating higher numbers of PBMC, addition of IL-7, or a combination of these. Furthermore, we have experimentally verified that the number of replicates needed for detection of weak responses can be calculated using parametric statistics. PMID:25738924

  15. Symptomatic Illness and Low CD4 Cell Count at HIV Seroconversion as Markers of Severe Primary HIV Infection

    PubMed Central

    Lodi, Sara; Fisher, Martin; Phillips, Andrew; De Luca, Andrea; Ghosn, Jade; Malyuta, Ruslan; Zangerle, Robert; Moreno, Santiago; Vanhems, Philippe; Boufassa, Faroudy; Guiguet, Marguerite; Porter, Kholoud

    2013-01-01

    Background The risk/benefit of initiating ART in primary HIV infection (PHI) is unclear. The benefits are more likely to outweigh the risks in patients with severe PHI. An accepted definition of severe PHI is, however, lacking. Methods CASCADE patients with HIV test interval <6 months were classified as severe and non-severe PHI based on whether the following traits were recorded in the first 6 months following seroconversion: severe specific pre-defined symptoms, central nervous system-implicated illness, and ?1, ?2 CD4<350 (and <500) cells/mm3. For each definition, we used Kaplan-Meier curves and Cox survival models to compare time to AIDS/death, censoring at the earlier of last clinic visit or 1/1/1997, when combination antiretroviral therapy (cART) became available. Results Among 1108 included patients mostly males (85%) infected through sex between men (71%), 366 were diagnosed with AIDS/died. The risk of AIDS/death was significantly higher for individuals with severe symptoms, those with ?1 CD4<350 cells/mm3 or ?2 CD4 <500 cells/mm3 in the first 6 months [aHR (95% confidence interval) 2.1 (1.4,3.2), 2.0 (1.5,2.7), and 2.3, (1.5–3.5) respectively]. Median [interquantile range] survival for patients with ?2, ?1 and no CD4<350 cells/mm3 within 6 months of seroconversion was 3.9 [2.7,6.5], 5.4 [4.5,8.4] and 8.1 [4.3,10.3] years, respectively. The diagnosis of CNS-implicated symptoms was rare and did not appear to be prognostic. Conclusion One CD4 count <350 or two <500 cells/mm3 within 6 months of seroconversion and/or severe illness in PHI may be useful early indicators of individuals at high risk of disease progression. PMID:24244330

  16. Fossomatic cell-counting on ewe milk: comparison with direct microscopy and study of variation factors.

    PubMed

    Gonzalo, C; Martínez, J R; Carrledo, J A; San Primitivo, F

    2003-01-01

    Using the Fossomatic method (FSCC) a total of 23,003 analytical SCC observations were carried out on 6400 aliquots taken from 80 individual ewe milk samples with the objective of studying the influence of 4 preservation procedures (without preservation, potassium dichromate, azidiol, and bronopol), 2 storage temperatures (ambient and refrigeration), 10 milk ages (3,6,12, and 24h, and 2,3,4,5,7, and 9d postcollection), and two analytical temperatures (40 and 60 degrees C). In addition, each sample was analyzed with direct microscopic method (DMSCC), using 3 different stainings for each sample: methylene blue (MB), May-Grünwald-Giemsa (MGG) and Pyronin Y-methyl green (PGM). This allowed DMSCC and FSCC (at 24 h of age) to be compared. The reference DMSCC from MB staining was a reliable method in ewe milk, though more specific stainings such as MGG and PMG slightly improve the residual standard deviation for repeated SCC. Between DMSCC and FSCC, the highest coefficients of correlation (0.972 to 0.996) corresponded to preserved and refrigerated milk, and the lowest (0.708 to 0.919) to unpreserved and ambient stored aliquots. Except for the unpreserved and ambient stored aliquots, SCC values were similar in all aliquots. Under FSCC, preservation, storage and analytical temperature, milk age, and most of the interactions showed a significant effect on SCC variation. In preserved samples, logSCC values ranged between 5.67 (bronopol) and 5.62 (azidiol). The higest values (5.72) were for unpreserved milk, which showed false overestimation of SCC due to bacterial proliferation. LogSCC was higher at 60 degrees C (5.68) than at 40 degrees C (5.65). The interaction between age, preservation and storage temperature showed no cell degeneration in properly handled samples over the 9 d of study. PMID:12613858

  17. COMPUTATIONAL FLUID DYNAMICS MODELING OF SOLID OXIDE FUEL CELLS

    E-print Network

    -dimensional model has been developed to simulate solid oxide fuel cells (SOFC). The model fully couples. The developed model is a full cell model, including all components of SOFC, flow channels, active and inactive. It is found that mass transfer limitation plays an important role in SOFC performance, especially under high

  18. Rate and predictors of non-AIDS events in a cohort of HIV-infected patients with a CD4 T cell count above 500 cells/mm³.

    PubMed

    Lucero, Constanza; Torres, Berta; León, Agathe; Calvo, Marta; Leal, Lorna; Pérez, Iñaki; Plana, Montserrat; Arnedo, Mireia; Mallolas, Josep; Gatell, Josep M; García, Felipe

    2013-08-01

    The reduction of risk of non-AIDS events after combined antiretroviral therapy (cART) initiation and the crude incidence rate (CIR) of these events in patients who control the viral load without cART (controllers) in a cohort of 574 antiretroviral-naive patients with a baseline CD4 T cell count above 500 cells/mm³ were assessed. Non-AIDS severe events were defined as a first admission to the hospital due to non-AIDS-defining malignancies, cardiovascular, neuropsychiatric, liver-related, or end-stage renal disease events. Potential determinants of non-AIDS/death events were studied using Cox regression models. Eighty-five non-AIDS/death events occurred during 6,062 persons-years of follow-up (PYFU) with a CIR of 1.4 per 100 PYFU. Factors associated with non-AIDS/death event were age (HR 3.4; 95% CI: 1.6-6.9), nadir CD4 below 350 cells/mm³ (HR 2.5; 95% CI: 1.4-4.6), and a last determination of viral load above the median (HR 1.9; 95% CI: 1.0-3.3). The CIR of non-AIDS/death events was 2.1 and 1.8 per 100 PYFU before and after cART in patients who started cART (n=446). A reduction of CIR of non-AIDS events after cART initiation was observed only in patients with a nadir of CD4 above 350 cells/mm³ (2.5 vs. 0.6 per 100 PYFU, p=0.004, and remained stable after cART in patients with a median nadir of CD4 below 350 cells/mm³. CIR was similar in elite, viremic, and noncontrollers (1.1, 1.0, and 1.5 per 100 PYFU, respectively, p=0.25). Reduction of CIR of non-AIDS events after cART initiation depends on nadir CD4 T cell count. Most of the controllers patients had a CIR similar to noncontrollers. These data support the early initiation of cART in HIV-infected patients. PMID:23530980

  19. C-reactive protein and white blood cell count as triage test between urgent and nonurgent conditions in 2961 patients with acute abdominal pain.

    PubMed

    Gans, Sarah L; Atema, Jasper J; Stoker, Jaap; Toorenvliet, Boudewijn R; Laurell, Helena; Boermeester, Marja A

    2015-03-01

    The purpose of this article is to assess the diagnostic accuracy of C-reactive protein (CRP) and white blood cell (WBC) count to discriminate between urgent and nonurgent conditions in patients with acute abdominal pain at the emergency department, thereby guiding the selection of patients for immediate diagnostic imaging.Data from 3 large published prospective cohort studies of patients with acute abdominal pain were combined in an individual patient data meta-analysis. CRP levels and WBC counts were compared between patients with urgent and nonurgent final diagnoses. Parameters of diagnostic accuracy were calculated for clinically applicable cutoff values of CRP levels and WBC count, and for combinations.A total of 2961 patients were included of which 1352 patients (45.6%) had an urgent final diagnosis. The median WBC count and CRP levels were significantly higher in the urgent group than in the nonurgent group (12.8?×10/L; interquartile range [IQR] 9.9-16) versus (9.3?×10/L; IQR 7.2-12.1) and (46? mg/L; IQR 12-100 versus 10 ?mg/L; IQR 7-26) (P?50 ?mg/L and WBC count >15?×10/L were combined; however, 85.3% of urgent cases was missed.A high CRP level (>50? mg/L) combined with a high WBC count (>15?×10/L) leads to the highest PPV. However, this applies only to a small subgroup of patients (8.7%). Overall, CRP levels and WBC count are insufficient markers to be used as a triage test in the selection for diagnostic imaging, even with a longer duration of complaints (>48 ?hours). PMID:25738473

  20. Electrical conductivity measurements of aqueous fluids under pressure with a hydrothermal diamond anvil cell.

    PubMed

    Ni, Huaiwei; Chen, Qi; Keppler, Hans

    2014-11-01

    Electrical conductivity data of aqueous fluids under pressure can be used to derive the dissociation constants of electrolytes, to assess the effect of ionic dissociation on mineral solubility, and to interpret magnetotelluric data of earth's interior where a free fluid phase is present. Due to limitation on the tensile strength of the alloy material of hydrothermal autoclaves, previous measurements of fluid conductivity were mostly restricted to less than 0.4 GPa and 800?°C. By adapting a Bassett-type hydrothermal diamond anvil cell, we have developed a new method for acquiring electrical conductivity of aqueous fluids under pressure. Our preliminary results for KCl solutions using the new method are consistent with literature data acquired with the conventional method, but the new method has great potential for working in a much broader pressure range. PMID:25430149

  1. Efficacy of standard vs. extended intramammary cefquinome treatment of clinical mastitis in cows with persistent high somatic cell counts.

    PubMed

    Swinkels, Jantijn M; Krömker, Volker; Lam, Theo J G M

    2014-11-01

    Extended duration of clinical mastitis (CM) treatment has been advocated, although results showing its higher efficacy compared with standard treatment are difficult to compare and seem conflicting. In a non-blinded, positively controlled clinical trial with systematic allocation, the efficacy of a standard, 1·5-d cefquinome treatment (ST), and an extended, 5-d intramammary cefquinome treatment (ET) were evaluated. The latter is frequently performed in cows with persistent high somatic cell count (SCC), expecting a better cure. Therefore, cows with CM immediately preceded by at least two consecutive monthly elevated SCC >200 000 cells/ml, were studied. The primary efficacy criteria were bacteriological cure (BC) and clinical cure (CC), while SCC cure was considered a secondary criterion of cure. Least square means of overall BC were not different after ET (79%, n=206) compared with ST (72%, n=203). ET, as compared with ST, improved BC of CM when caused by streptococci, specifically Streptococcus uberis. At day 1·5, only 13% of quarters showed CC, increasing significantly towards 60% at day 5, and 99% at day 14 and at day 21. No significant difference in CC was present between treatment groups. Overall SCC cure was low (22%) and not significantly different between treatment groups, but significantly higher for cases due to enterobacteriacae compared with staphylococci. In conclusion, ET with cefquinome of CM in cows with a persistent high SCC seems to be only indicated when caused by streptococci, mainly Str. uberis but shows no advantage when no information on bacteriological causes of mastitis is available. In our data, absence of CC directly after ST was not related to eventual BC. PMID:25230074

  2. A Novel Marker for Screening Paroxysmal Nocturnal Hemoglobinuria Using Routine Complete Blood Count and Cell Population Data

    PubMed Central

    Kahng, Jimin; Kim, Yonggoo; Kim, Jung Ok; Koh, Kwangsang; Lee, Jong Wook

    2015-01-01

    Background Final diagnosis of paroxysmal nocturnal hemoglobinuria (PNH) may take years demanding a quick diagnosis measure. We used the facts that PNH cells are damaged in acid, and reagents for measuring reticulocytes in Coulter DxH800 (Beckman Coulter, USA) are weakly acidic and hypotonic, to create a new PNH screening marker. Methods We analyzed 979 complete blood counts (CBC) data from 963 patients including 57 data from 44 PNH patients. Standard criteria for PNH assay for population selection were followed: flow cytometry for CD55 and CD59 on red blood cells (RBCs) to a detection level of 1%; and fluorescent aerolysin, CD24 and CD15 in granulocytes to 0.1%. Twenty-four PNH minor clone-positive samples (minor-PNH+) were taken, in which the clone population was <5% of RBCs and/or granulocytes. Excluding PNH and minor-PNH+ patients, the population was divided into anemia, malignancy, infection, and normal groups. Parameters exhibiting a distinct demarcation between PNH and non-PNH groups were identified, and each parameter cutoff value was sought that includes the maximum [minimum] number of PNH [non-PNH] patients. Results Cutoff values for 5 selected CBC parameters (MRV, RDWR, MSCV, MN-AL2-NRET, and IRF) were determined. Positive rates were: PNH (86.0%), minor-PNH+ (33.3%), others (5.0%), anemia (13.4%), malignancy (5.3%), infection (3.7%), normal (0.0%); within anemia group, aplastic anemia (40.0%), immune hemolytic anemia (11.1%), iron deficiency anemia (1.6%). Sensitivity (86.0%), specificity (95.0%), PPV (52.1%), and NPV (99.1%) were achieved in PNH screening. Conclusion A new PNH screening marker is proposed with 95% specificity and 86% sensitivity. The flag identifies PNH patients, reducing time to final diagnosis by flow cytometry. PMID:25553278

  3. Analysis of changes in the total lymphocyte and eosinophil count during immunotherapy for metastatic renal cell carcinoma: correlation with response and survival.

    PubMed

    Jeong, In Gab; Han, Kyung Seok; Joung, Jae Young; Choi, Woo Suk; Hwang, Seung Sik; Yang, Seung Ok; Seo, Ho Kyung; Chung, Jinsoo; Lee, Kang Hyun

    2007-09-01

    The aims of this study were to analyze lymphocyte and eosinophil counts in consecutive peripheral blood samples taken during immunotherapy for metastatic renal cell carcinoma (mRCC) and to correlate the findings with objective response and survival. A total of 40 patients with mRCC who received immunotherapy with interleukin-2, interferon-alpha, and 5-fluorouracil were analyzed. Objective responses were observed in 14 patients, including 2 (5%) who showed a complete response (CR) and 12 (30%) who showed a partial response (PR). Eleven patients (27%) achieved stable disease (SD), and 15 patients (38%) had progressive disease (PD). Changes from baseline in the total lymphocyte counts were significantly higher in the responding patients (CR+PR+SD) than in the non-responding patients (PD) (p=0.017), but no difference was seen in the total eosinophil counts (p=0.275). Univariate analysis identified the Eastern Cooperative Oncology Group (ECOG) performance status (p=0.017), the presence of a primary renal tumor (p<0.001) and the peripheral lymphocyte counts at week 4 (p=0.034) as prognostic factors, but a low ECOG performance status (p=0.003) and the presence of a primary renal tumor (p=0.001) were identified as independent poor prognostic factors by multivariate analysis. This study provides further evidence that changes in blood lymphocyte counts may serve as an objective indicator of objective responses. PMID:17923738

  4. Exploring the characteristics and dynamics of Ontario dairy herds experiencing increases in bulk milk somatic cell count during the summer.

    PubMed

    Shock, D A; LeBlanc, S J; Leslie, K E; Hand, K; Godkin, M A; Coe, J B; Kelton, D F

    2015-06-01

    Regionally aggregated bulk milk somatic cell count (BMSCC) data from around the world shows a repeatable cyclicity, with the highest levels experienced during warm, humid seasons. No studies have evaluated this seasonal phenomenon at the herd level. The objectives of this study were to define summer seasonality in BMSCC on an individual herd basis, and subsequently to describe the characteristics and dynamics of herds with increased BMSCC in the summer. The data used for this analysis were from all dairy farms in Ontario, Canada, between January 2000 and December 2011 (n ? 4,000 to 6,000 herds/yr). Bulk milk data were obtained from the milk marketing board and consisted of bulk milk production, components (fat, protein, lactose, other solids), and quality (BMSCC, bacterial count, inhibitor presence, freezing point), total milk quota of the farm, and milk quota and incentive fill percentage. A time-series linear mixed model, with random slopes and intercepts, was constructed using sine and cosine terms as predictors to describe seasonality, with herd as a random effect. For each herd, seasonality was described with reference to 1 cosine function of variable amplitude and phase shift. The predicted months of maximal and minimal BMSCC were then calculated. Herds were assigned as low, medium, and high summer increase (LSI, MSI, and HSI, respectively) based on percentiles of amplitude in BMSCC change for each of the 4 seasons. Using these seasonality classifications, 2 transitional repeated measures logistic regression models were built to assess the characteristics of MSI and HSI herds, using LSI herds as controls. Based on the analyses performed, a history of summer BMSCC increases increased the odds of experiencing a subsequent increase. As herd size decreased, the odds of experiencing HSI to MSI in BMSCC increased. Herds with more variability in daily BMSCC were at higher odds of experiencing MSI and HSI in BMSCC, as were herds with lower annual mean BMSCC. Finally, a negative association was noted between filling herd production targets and experiencing MSI to HSI in BMSCC. These findings provide farm advisors direction for predicting herds likely to experience increases in SCC over the summer, allowing them to proactively focus udder health prevention strategies before the high-risk summer period. PMID:25864052

  5. Antibody-Functionalized Fluid-Permeable Surfaces for Rolling Cell Capture at High Flow Rates

    PubMed Central

    Mittal, Sukant; Wong, Ian Y.; Deen, William M.; Toner, Mehmet

    2012-01-01

    Adhesion-based cell capture on surfaces in microfluidic devices forms the basis of numerous biomedical diagnostics and in vitro assays. However, the performance of these platforms is partly limited by interfacial phenomena that occur at low Reynolds numbers. In contrast, cell homing to porous vasculature is highly effective in vivo during inflammation, stem cell trafficking, and cancer metastasis. Here, we show that a porous, fluid-permeable surface functionalized with cell-specific antibodies promotes efficient and selective cell capture in vitro. This architecture is advantageous due to enhanced transport as streamlines are diverted toward the surface. Moreover, specific cell-surface interactions are promoted due to reduced shear, allowing gentle cell rolling and arrest. Together, these synergistic effects enable highly effective cell capture at flow rates more than an order of magnitude larger than those provided by existing devices with solid surfaces. PMID:22385842

  6. Therapeutic outcomes of transplantation of amniotic fluid-derived stem cells in experimental ischemic stroke.

    PubMed

    Tajiri, Naoki; Acosta, Sandra; Portillo-Gonzales, Gabriel S; Aguirre, Daniela; Reyes, Stephanny; Lozano, Diego; Pabon, Mibel; Dela Peña, Ike; Ji, Xunming; Yasuhara, Takao; Date, Isao; Solomita, Marianna A; Antonucci, Ivana; Stuppia, Liborio; Kaneko, Yuji; Borlongan, Cesar V

    2014-01-01

    Accumulating preclinical evidence suggests the use of amnion as a source of stem cells for investigations of basic science concepts related to developmental cell biology, but also for stem cells' therapeutic applications in treating human disorders. We previously reported isolation of viable rat amniotic fluid-derived stem (AFS) cells. Subsequently, we recently reported the therapeutic benefits of intravenous transplantation of AFS cells in a rodent model of ischemic stroke. Parallel lines of investigations have provided safety and efficacy of stem cell therapy for treating stroke and other neurological disorders. This review article highlights the need for investigations of mechanisms underlying AFS cells' therapeutic benefits and discusses lab-to-clinic translational gating items in an effort to optimize the clinical application of the cell transplantation for stroke. PMID:25165432

  7. Therapeutic outcomes of transplantation of amniotic fluid-derived stem cells in experimental ischemic stroke

    PubMed Central

    Tajiri, Naoki; Acosta, Sandra; Portillo-Gonzales, Gabriel S.; Aguirre, Daniela; Reyes, Stephanny; Lozano, Diego; Pabon, Mibel; Dela Peña, Ike; Ji, Xunming; Yasuhara, Takao; Date, Isao; Solomita, Marianna A.; Antonucci, Ivana; Stuppia, Liborio; Kaneko, Yuji; Borlongan, Cesar V.

    2014-01-01

    Accumulating preclinical evidence suggests the use of amnion as a source of stem cells for investigations of basic science concepts related to developmental cell biology, but also for stem cells’ therapeutic applications in treating human disorders. We previously reported isolation of viable rat amniotic fluid-derived stem (AFS) cells. Subsequently, we recently reported the therapeutic benefits of intravenous transplantation of AFS cells in a rodent model of ischemic stroke. Parallel lines of investigations have provided safety and efficacy of stem cell therapy for treating stroke and other neurological disorders. This review article highlights the need for investigations of mechanisms underlying AFS cells’ therapeutic benefits and discusses lab-to-clinic translational gating items in an effort to optimize the clinical application of the cell transplantation for stroke. PMID:25165432

  8. Effect of Varying Fluid Shear Stress on Cancer Stem Cell Viability & Protein Expression

    NASA Astrophysics Data System (ADS)

    Domier, Ria; Kim, Yonghyun; Dozier, David; Triantafillu, Ursula

    2013-11-01

    Cancer stem cells cultured in vitro in stirred bioreactors are exposed to shear stress. By observing the effect of shear stress on cancer stem cell viability, laboratory cell growth could be optimized. In addition, metastasized cancer stem cells in vivo are naturally exposed to shear stress, a factor influencing stem cell differentiation, while circulating in the bloodstream. Changes in protein expression after exposure to shear stress could allow for identification and targeting of circulating cancer cells. In this study, blood flow through capillaries was simulated by using a syringe pump to inject suspensions of Kasumi-1 leukemia stem cells into model blood vessels composed of PEEK tubing 125 microns in diameter. The Hagen-Poisseuille equation was used to solve for operating flow rates based on specified amounts of shear stress. After exposure, cell counts and viabilities were observed using an optical microscope and proteins were analyzed using Western blotting. It was observed that at a one minute exposure to stress, cell viability increased as the amount of shear was increased from 10 to 60 dynes per square centimeter. Results from this research are applicable to optimization of large-scale stem cell growth in bioreactors as well as to the design of targeted cancer therapies. Cancer stem cells cultured in vitro in stirred bioreactors are exposed to shear stress. By observing the effect of shear stress on cancer stem cell viability, laboratory cell growth could be optimized. In addition, metastasized cancer stem cells in vivo are naturally exposed to shear stress, a factor influencing stem cell differentiation, while circulating in the bloodstream. Changes in protein expression after exposure to shear stress could allow for identification and targeting of circulating cancer cells. In this study, blood flow through capillaries was simulated by using a syringe pump to inject suspensions of Kasumi-1 leukemia stem cells into model blood vessels composed of PEEK tubing 125 microns in diameter. The Hagen-Poisseuille equation was used to solve for operating flow rates based on specified amounts of shear stress. After exposure, cell counts and viabilities were observed using an optical microscope and proteins were analyzed using Western blotting. It was observed that at a one minute exposure to stress, cell viability increased as the amount of shear was increased from 10 to 60 dynes per square centimeter. Results from this research are applicable to optimization of large-scale stem cell growth in bioreactors as well as to the design of targeted cancer therapies. Funding from NSF REU grant #1062611 is gratefully acknowledged.

  9. Consequence of alternative standards for bulk tank somatic cell count of dairy herds in the United States.

    PubMed

    Norman, H D; Lombard, J E; Wright, J R; Kopral, C A; Rodriguez, J M; Miller, R H

    2011-12-01

    Noncompliance with current US and European Union (EU) standards for bulk-tank somatic cell count (BTSCC) as well as BTSCC standards recently proposed by 3 US organizations was evaluated using US Dairy Herd Improvement Association (DHI) herds and herds supplying milk to 4 Federal Milk Marketing Orders (FMO). Herds with 15 to 26 tests (frequently monthly) from January 2009 through October 2010 were included. Somatic cell scores (SCS) from 14,854 herds and 164,794 herd-tests were analyzed for DHI herds with ?10 cows for all tests. Herd test-day SCC was derived as a proxy for BTSCC and was the basis for determining noncompliance and percentage of the milk it represented. For FMO herds, actual milk marketed and BTSCC were available from 27,759 herds and 325,690 herd-tests. A herd was noncompliant for the current EU BTSCC standard after 4 consecutive rolling 3-test geometric means (geometric method) were >400,000 cells/mL. A herd was noncompliant for the current US BTSCC standard after 3 of 5 consecutive monthly BTSCC shipments (frequency method) were >750,000 cells/mL. Alternative proposed standards (600,000, 500,000, or 400,000 cells/mL) also were examined. A third method designated noncompliance when a single 3-mo geometric mean of >550,000 or >400,000 cells/mL and a subsequent test exceeded the same level. Results were examined based on herd size or milk shipped by month. Noncompliance for the current US standard for the 12 mo ending October 2010 in DHI and FMO herds was 0.9 and 1.0%, respectively, compared with 7.8 and 16.1% for the current EU standard. Noncompliance was always greater for the frequency method than for the geometric method and was inversely related to herd size or milk shipped. Using the frequency method at 400,000 cells/mL, noncompliance was 19.1% for DHI herd-tests in herds with <50 cows compared with 1.1% for herds with ? 1,000 cows. For FMO herds shipping <900 t, noncompliance was 44.5% using the frequency method at 400,000 cells/mL compared with 8.0% for herds marketing >9,000 t. All methods proposed increased the percentages of herds and shipped milk that exceeded the regulatory limit. Producers will need to place more emphasis on reducing the incidence and prevalence of subclinical mastitis through known management practices such as proper milking techniques, well-functioning milking machines, postmilking teat disinfectant, dry cow treatment, and culling of problem cows to meet any of the proposed new standards. PMID:22118112

  10. Counting Penguins.

    ERIC Educational Resources Information Center

    Perry, Mike; Kader, Gary

    1998-01-01

    Presents an activity on the simplification of penguin counting by employing the basic ideas and principles of sampling to teach students to understand and recognize its role in statistical claims. Emphasizes estimation, data analysis and interpretation, and central limit theorem. Includes a list of items for classroom discussion. (ASK)

  11. Biodiversity Count

    NSDL National Science Digital Library

    Suzanne Savanick, Science Education Resource Center, Carleton College, ssavanic@carleton.edu

    In this class exercise, students count the number of species they can find in a five minute block of time in both an urban lawn and natural, remnant forest area. The students are introduced to the concept of low and high biodiversity areas and engage in a discussion about biodiversity loss.

  12. Counting Populations

    ERIC Educational Resources Information Center

    Damonte, Kathleen

    2004-01-01

    Scientists use sampling to get an estimate of things they cannot easily count. A population is made up of all the organisms of one species living together in one place at the same time. All of the people living together in one town are considered a population. All of the grasshoppers living in a field are a population. Scientists keep track of the…

  13. Counting Coins

    NSDL National Science Digital Library

    K12, Inc.

    2011-03-23

    In this iOS app students practice counting U.S. coins by matching the value, making the total, telling how much, and creating their own values. Students drag coins onto a digital mat or enter values with a keypad to complete the tasks, and then receive feedback.

  14. Increased S100B+ NK cell counts in acutely ill schizophrenia patients are correlated with the free cortisol index, but not with S100B serum levels.

    PubMed

    Steiner, Johann; Westphal, Sabine; Schroeter, Matthias L; Schiltz, Kolja; Jordan, Wolfgang; Müller, Ulf J; Bernstein, Hans-Gert; Bogerts, Bernhard; Schmidt, Reinhold E; Jacobs, Roland

    2012-05-01

    Several studies have provided evidence for increased S100B serum concentrations in schizophrenia. The pathophysiological significance of this finding is still uncertain because S100B is involved in many cellular mechanisms and is not astrocyte-specific as was previously assumed. S100B is also expressed by subsets of CD3+ CD8+ T cells and natural killer (NK) cells and may therefore be linked to the immune hypothesis of schizophrenia. We have quantified S100B+ CD3+ CD8+ T cells and NK cells by flow cytometry in the peripheral blood of 26 acutely ill schizophrenia cases and 32 matched controls. In parallel, S100B concentrations and the free cortisol index (FCI), a surrogate marker for stress axis activity, were determined in serum samples from the same blood draw. Psychopathology was monitored using the Positive and Negative Syndrome Scale (PANSS). The patient group had increased S100B+ NK cell counts (P=0.045), which correlated with the FCI (r=0.299, P=0.026) but not with the PANSS or the elevated (P=0.021) S100B serum concentrations. S100B+ CD3+ CD8+ T cell counts were not significantly changed in the patient group and did neither correlate with the FCI and PANSS, nor with S100B serum concentrations. In conclusion, despite the observation of an increase in S100B+ NK cells in schizophrenia patients, the lack of a correlation with serum S100B concentrations suggests that these cells are probably not a major source of S100B in the blood of schizophrenia patients. Notably, elevated S100B+ NK cell counts may be linked with stress axis activation. PMID:22326439

  15. Effect of cell phone magnetic fields on adjustable cerebrospinal fluid shunt valves

    Microsoft Academic Search

    Sadahiro Nomura; Hirosuke Fujisawa; Michiyasu Suzuki

    2005-01-01

    The rapid increase in the number of cell phone users has led to the suggestion that electromagnetic waves might affect medical devices. Cerebrospinal fluid shunt valves contain a magnetic device to allow the intracranial pressure setting to be adjusted transcutaneously. Among the valves tested, the settings of the Strata valve, the Hakim valve, and the Sophy valve were affected by

  16. Computational fluid dynamics modeling of a solid oxide electrolyzer cell for hydrogen production

    Microsoft Academic Search

    Meng Ni

    2009-01-01

    A 2D computational fluid dynamics (CFD) model was developed to study the performance of a planar solid oxide electrolyzer cell (SOEC) for hydrogen production. The governing equations for mass continuity, momentum conservation, energy conservation and species conservation were discretized with the finite volume method (FVM). The coupling of velocity and pressure was treated with the SIMPLEC (Semi-Implicit Method for Pressure

  17. Impact of Baseline HIV-1 Tropism on Viral Response and CD4 Cell Count Gains in HIV-Infected Patients Receiving First-line Antiretroviral Therapy

    PubMed Central

    Seclén, Eduardo; Soriano, Vicente; González, María M.; Martín-Carbonero, Luz; Gellermann, Holger; Distel, Manuel; Kadus, Werner

    2011-01-01

    Background.?Viral tropism influences the natural history of human immunodeficiency type 1 (HIV-1) disease: X4 viruses are associated with faster decreases in CD4 cell count. There is scarce information about the influence of viral tropism on treatment outcomes. Methods.?Baseline plasma samples from patients recruited to the ArTEN (Atazanavir/ritnoavir vs. Nevirapine on a background of Tenofovir and Emtricitabine) trial were retrospectively tested for HIV-1 tropism using the genotypic tool geno2phenoFPR=5.75%. ArTEN compared nevirapine with atazanavir-ritonavir, both along with tenofovir-emtricitabine, in drug-naïve patients. Results.?Of 569 ArTEN patients, 428 completed 48 weeks of therapy; 282 of these received nevirapine and 146 of these received atazanavir-ritonavir. Overall, non-B subtypes of HIV-1 were recognized in 96 patients (22%) and X4 viruses were detected in 55 patients (14%). At baseline, patients with X4 viruses had higher plasma HIV RNA levels (5.4 vs 5.2 log copies/mL, respectively; P = .044) and lower CD4 cell counts (145 vs 188 cells/?L, respectively; P < .001) than those with R5 strains. At week 48, virologic responses were lower in patients with X4 viruses than in patients with R5 viruses (77% vs 92%, respectively; P = .009). Multivariate analysis confirmed HIV-1 tropism as an independent predictor of virologic response at week 24 (P = .012). This association was extended to week 48 (P = .007) in clade B viruses. Conversely, CD4 cell count recovery was not influenced by baseline HIV-1 tropism. Conclusions.?HIV-1 tropism is an independent predictor of virologic response to first-line antiretroviral therapy. In contrast, it does not seem to influence CD4 cell count recovery. Clinical Trials Registration.?NCT00389207. PMID:21628668

  18. Fluid Flow through a High Cell Density Fluidized-Bed during Centrifugal Bioreactor Culture

    PubMed Central

    Detzel, Christopher J.; Van Wie, Bernard J.; Ivory, Cornelius F.

    2010-01-01

    An increasing demand for products such as tissues, proteins, and antibodies from mammalian cell suspension cultures is driving interest in increasing production through high-cell density bioreactors. The centrifugal bioreactor (CCBR) retains cells by balancing settling forces with surface drag forces due to medium throughput and is capable of maintaining cell densities above 108 cells/mL. This article builds on a previous study where the fluid mechanics of an empty CCBR were investigated showing fluid flow is nonuniform and dominated by Coriolis forces, raising concerns about nutrient and cell distribution. In this article, we demonstrate that the previously reported Coriolis forces are still present in the CCBR, but masked by the presence of cells. Experimental dye injection observations during culture of 15 ?m hybridoma cells show a continual uniform darkening of the cell bed, indicating the region of the reactor containing cells is well mixed. Simulation results also indicate the cell bed is well mixed during culture of mammalian cells ranging in size from 10 to 20 ?m. However, simulations also allow for a slight concentration gradient to be identified and attributed to Coriolis forces. Experimental results show cell density increases from 0.16 to 0.26 when centrifugal force is doubled by increasing RPM from 650 to 920 at a constant inlet velocity of 6.5 cm/s; an effect also observed in the simulation. Results presented in this article indicate cells maintained in the CCBR behave as a high-density fluidized bed of cells providing a homogeneous environment to ensure optimal growth conditions. PMID:20205172

  19. Obstructive renal injury: from fluid mechanics to molecular cell biology

    PubMed Central

    Ucero, Alvaro C; Gonçalves, Sara; Benito-Martin, Alberto; Santamaría, Beatriz; Ramos, Adrian M; Berzal, Sergio; Ruiz-Ortega, Marta; Egido, Jesus; Ortiz, Alberto

    2010-01-01

    Urinary tract obstruction is a frequent cause of renal impairment. The physiopathology of obstructive nephropathy has long been viewed as a mere mechanical problem. However, recent advances in cell and systems biology have disclosed a complex physiopathology involving a high number of molecular mediators of injury that lead to cellular processes of apoptotic cell death, cell injury leading to inflammation and resultant fibrosis. Functional studies in animal models of ureteral obstruction using a variety of techniques that include genetically modified animals have disclosed an important role for the renin-angiotensin system, transforming growth factor-?1 (TGF-?1) and other mediators of inflammation in this process. In addition, high throughput techniques such as proteomics and transcriptomics have identified potential biomarkers that may guide clinical decision-making. PMID:24198613

  20. Association between somatic cell count early in the first lactation and the longevity of Irish dairy cows.

    PubMed

    Archer, S C; Mc Coy, F; Wapenaar, W; Green, M J

    2013-05-01

    Reduced longevity of cows is an important component of mastitis costs, and increased somatic cell count (SCC) early in the first lactation has been reported to increase culling risk throughout the first lactation. Generally, cows must survive beyond the first lactation to break even on their rearing costs. The aim of this research was to assess the association between SCC of primiparous cows at 5 to 30 days in milk (SCC1), and survival over a 5-y period for cows in Irish dairy herds. The data set used for model development was based on 147,458 test day records from 7,537 cows in 812 herds. Cows were censored at their last recording if identified at a later date in other herds or if recorded at the last available test date for their herd, otherwise, date of disposal was taken to be at the last test date for each cow. Survival time was calculated as the number of days between the dates of first calving and the last recording, which was split into 50-d intervals. Data were analyzed in discrete time logistic survival models that accounted for clustering of 50-d intervals within cows, and cows within herds. Models were fitted in a Bayesian framework using Markov chain Monte Carlo simulations. Model fit was assessed by comparison of posterior predictions to the observed disposal risk for cows aggregated by parameters in the model. Model usefulness was assessed by cross validation in a separate data set, which contained 144,113 records from 7,353 cows in 808 herds, and posterior predictions were compared with the observed disposal risk for cows aggregated by parameters of biological importance. Disposal odds increased by a factor of 5% per unit increase in ln SCC1. Despite this, posterior predictive distributions revealed that the probability of reducing replacement costs by >€10 per heifer calved, through decreasing the herd level prevalence of cows with SCC1 ? 400,000 cells/mL (from an initial prevalence of ? 20 to <10%) only exceeded 50% for less than 1 in 5 Irish herds. These results indicate that the effect of a reduction in the prevalence of cows with SCC1 ? 400,000 cells/mL on replacement costs alone for most Irish dairy herds is small, and future research should investigate other potential losses, such as the effect of SCC1 on lifetime milk yield. PMID:23522676

  1. Investigating a dilution effect between somatic cell count and milk yield and estimating milk production losses in Irish dairy cattle.

    PubMed

    Boland, F; O'Grady, L; More, S J

    2013-03-01

    Increased somatic cell counts (SCC) are associated with reduced milk yield. Additionally, it has been hypothesized that as milk yield increases, SCC is diluted in cattle without an intramammary infection (IMI). If the hypothesis is correct, estimates of SCC from high-yielding cattle without an IMI are likely to be lower than those from low-yielding cattle without an IMI. The objectives of this paper were to investigate the presence of a potential dilution effect between SCC and milk yield, overall and by parity, and to estimate lactation milk production losses with increasing SCC in Irish dairy cattle. The data consisted of 100 randomly selected herds from all milk recording herds between 2008 and 2010. The data set comprised 8,229 cows, of which approximately 90% were Holstein or Holstein crossbred animals. Various adjustments were used to investigate the presence of a potential dilution effect between SCC and milk yield; additionally, lactation milk production losses with increasing SCC and parity were estimated. The data had an inherent hierarchical structure, with lactations nested within cows and cows within herds; thus, a linear mixed model with 2 random effects was used. We found no evidence of a dilution effect of SCC with increasing milk yield in Irish dairy cattle. Average milk production losses were estimated, and they increased with increasing SCC compared with the referent of ? 50,000 cells/mL. Additionally, for all SCC values for parities 1 to 3, this production loss increased significantly with increasing parity. Estimated milk losses for parities 4 and 5 did not differ, and differences between parities 3 and 4 were significant only for SCC values <300,000 cells/mL. The estimated milk loss with increasing SCC varies greatly across studies, with the results from the current study exceeding most previously published results (except for results from the UK). Several factors could explain these differences, including geographic factors such as milk yield and predominant mastitis pathogens. The dilution effect warrants further work, as does the effect of prior duration of increased SCC on milk yield and the potential for compensation of milk yield losses over a lactation. PMID:23295120

  2. Fluid and cell behaviors along a 3D printed alginate/gelatin/fibrin channel.

    PubMed

    Xu, Yufan; Wang, Xiaohong

    2015-08-01

    Three-dimensional (3D) cell manipulation is available with the integration of microfluidic technology and rapid prototyping techniques. High-Fidelity (Hi-Fi) constructs hold enormous therapeutic potential for organ manufacturing and regenerative medicine. In the present paper we introduced a quasi-three-dimensional (Q3D) model with parallel biocompatible alginate/gelatin/fibrin hurdles. The behaviors of fluids and cells along the microfluidic channels with various widths were studied. Cells inside the newly designed microfluidic channels attached and grew well. Morphological changes of adipose-derived stem cells (ADSCs) in both two-dimensional (2D) and 3D milieu were found on the printed constructs. Endothelialization occurred with the co-cultures of ADSCs and hepatocytes. This study provides insights into the interactions among fluids, cells and biomaterials, the behaviors of fluids and cells along the microfluidic channels, and the applications of Q3D techniques. Biotechnol. Bioeng. 2015;112: 1683-1695. © 2015 Wiley Periodicals, Inc. PMID:25727058

  3. High interindividual variability in the CD4/CD8 T cell ratio and natalizumab concentration levels in the cerebrospinal fluid of patients with multiple sclerosis.

    PubMed

    Harrer, A; Pilz, G; Wipfler, P; Oppermann, K; Sellner, J; Hitzl, W; Haschke-Becher, E; Afazel, S; Rispens, T; van der Kleij, D; Trinka, E; Kraus, J

    2015-06-01

    Strongly decreased leucocyte counts and a reduced CD4/CD8 T cell ratio in the cerebrospinal fluid (CSF) of natalizumab (NZB)-treated multiple sclerosis (MS) patients may have implications on central nervous (CNS) immune surveillance. With regard to NZB-associated progressive multi-focal leucoencephalopathy, we aimed at delineating a relationship between free NZB, cell-bound NZB, adhesion molecule (AM) expression and the treatment-associated shift in the CSF T cell ratio. Peripheral blood (PB) and CSF T cells from 15 NZB-treated MS patients, and CSF T cells from 10 patients with non-inflammatory neurological diseases and five newly diagnosed MS patients were studied. Intercellular adhesion molecule-1 (ICAM-1), leucocyte function antigen-1 (LFA-1), very late activation antigen-4 (VLA-4), NZB saturation levels, and T cell ratios were analysed by flow cytometry. NZB concentrations were measured by enzyme-linked immunosorbent assay (ELISA). Lower NZB saturation levels (P?cells. CSF T cell ratios (0·3-2·1) and NZB concentrations (0·01-0·42 µg/ml) showed a pronounced interindividual variance. A correlation between free NZB, cell-bound NZB or AM expression levels and the CSF T cell ratio was not found. Extremely low NZB concentrations and a normalized CSF T cell ratio were observed in one case. The differential NZB saturation and AM expression of CSF CD8 T cells may contribute to their relative enrichment in the CSF. The reduced CSF T cell ratio appeared sensitive to steady-state NZB levels, as normalization occurred quickly. The latter may be important concerning a fast reconstitution of CNS immune surveillance. PMID:25603898

  4. Fluid Shear Stress Pre-Conditioning Promotes Endothelial Morphogenesis of Embryonic Stem Cells Within Embryoid Bodies

    PubMed Central

    Nsiah, Barbara A.; Ahsan, Tabassum; Griffiths, Sarah; Cooke, Marissa; Nerem, Robert M.

    2014-01-01

    Pluripotent embryonic stem cells (ESCs) are capable of differentiating into all mesoderm-derived cell lineages, including endothelial, hematopoietic, and cardiac cell types. Common strategies to direct mesoderm differentiation of ESCs rely on exposing the cells to a series of biochemical and biophysical cues at different stages of differentiation to promote maturation toward specific cell phenotypes. Shear forces that mimic cardiovascular physiological forces can evoke a myriad of responses in somatic and stem cell populations, and have, thus, been studied as a means to direct stem cell differentiation. However, elucidating the effects of shear pre-conditioning on the subsequent vascular differentiation and morphogenesis of ESCs has yet to be examined. In this study, ESC monolayers were subjected to physiological shear (5?dyn/cm2) or static conditions for 2 days on collagen IV-coated substrates before initiating embryoid body (EB) differentiation. Immediately after the pre-conditioning period, shear pre-conditioned and statically cultured ESCs exhibited similar morphologies and largely retained a pluripotent phenotype; however, ESCs exposed to fluid shear expressed increased levels of endothelial marker genes Flk-1 (?3-fold), VE-cadherin (?3-fold), and PECAM (?2-fold), compared with statically cultured ESCs. After 7 days of EB culture, ?70% of EBs formed from shear pre-conditioned ESCs expressed significantly higher levels of endothelial marker genes compared with EBs formed from statically cultured ESCs. Interestingly, unlike EBs formed from statically cultured ESCs, EBs formed from fluid shear stress pre-conditioned ESCs exhibited a centrally localized region of VE-cadherin+ cells that persisted for at least 10 days of differentiation. These results demonstrate that fluid shear stress pre-conditioning not only promotes ESC endothelial gene expression but also subsequently impacts the organization of endothelial cells within EBs. Together, these studies highlight a novel approach to promote in vitro morphogenesis of developmental vasculogenic models and potentially promote pre-vascularization of tissue-engineered constructs derived from pluripotent stem cells. PMID:24138406

  5. A mathematical model of fluid secretion from a parotid acinar cell

    PubMed Central

    Gin, Elan; Crampin, Edmund J.; Brown, David A.; Shuttleworth, Trevor J.; Yule, David I.; Sneyd, James

    2007-01-01

    Salivary fluid secretion is crucial for preventing problems such as dryness of mouth, difficulty with mastication and swallowing, as well as oral pain and dental cavities. Fluid flow is driven primarily by the transepithelial movement of chloride and sodium ions into the parotid acinus lumen. The activation of Cl? channels is calcium dependent, with the average elevated calcium concentration during calcium oscillations increasing the conductance of the channels, leading to an outflow of Cl?. The accumulation of NaCl in the lumen drives water flow by osmosis. We construct a mathematical model of the calcium concentration oscillations and couple this to a model for Cl? efflux. We also construct a model governing fluid flow in an isolated parotid acinar cell, which includes a description of the rate of change of intracellular ion concentrations, cell volume, membrane potential and water flow rate. We find that [Ca2+] oscillations lead to oscillations in fluid flow, and that the rate of fluid flow is regulated by the average calcium concentration and not the frequency of the oscillations. PMID:17559884

  6. Mastitis Is Associated with Increased Free Fatty Acids, Somatic Cell Count, and Interleukin-8 Concentrations in Human Milk

    PubMed Central

    Hunt, Katherine M.; Williams, Janet E.; Shafii, Bahman; Hunt, Martha K.; Behre, Rebecca; Ting, Robert; McGuire, Michelle K.

    2013-01-01

    Abstract Background Research in bovine lactation has demonstrated that milk produced by a mammary gland displaying inflammation-based symptoms of mastitis has increased levels of free fatty acids (FFAs) compared with milk produced by a contralateral asymptomatic gland. However, the effects of mastitis on lipid classes in milk have not been investigated in humans. Methods The study described here compared milk collected from the symptomatic breast of women with mastitis (n=14) with that collected from the contralateral asymptomatic breast to determine if mastitis caused alterations in the quantity of total lipids, FFAs, and phospholipids (PLs), as well as the fatty acid profiles of these lipid classes. To assess their efficacy as biomarkers of mastitis, samples were also analyzed for selected markers of local inflammation: sodium, somatic cell count (SCC), and interleukin-8 (IL-8). Results FFAs were higher in milk from the mastitic breast compared with that from the healthy breast (1.31 vs. 1.07±0.10?g/100?g of lipid, p<0.05). Similarly, SCC and IL-8 were elevated roughly 10-fold in milk from mastitic breasts, compared with milk from healthy breasts, and sodium tended to be higher in milk from mastitic breasts (p<0.10). However, there were no differences in total lipid, PLs, or fatty acid profiles within each lipid class. Conclusions In summary, mastitis is associated with increased lipolysis in the human breast but not alterations in milk fat synthesis, as evidenced by a lack of alteration in total milk lipids. Additionally, these results indicate that SCC and IL-8 may be better indicators of mammary inflammation than sodium content. PMID:22283504

  7. Factors associated with high milk test day somatic cell counts in large dairy herds in Brandenburg. II. Milking practices.

    PubMed

    Köster, G; Tenhagen, B-A; Scheibe, N; Heuwieser, W

    2006-05-01

    The objective of this study was to examine the influences of different milking practices on cow udder health in 80 large dairy herds (range 100-1100 cows) in Brandenburg, Germany. Milking practices were evaluated during one complete milking using a standardized data capture form. The somatic cell count (SCC) of all lactating cows on each farm was determined monthly by the local milk recording association 'Landeskontrollverband Brandenburg'. Factor analysis was used to investigate the relationship between the different aspects of the milking practices. The components extracted by the factor analysis were examined for their influence on the SCC of the current month (CMSCC) and the year before the visit (YASCC) using univariate analysis of variance. Three components were extracted from the milking practices. 'Reasonable use of water' was significantly related to CMSCC (P = 0.019) and YASCC (P = 0.003). It included information on the use of a hose to clean udders before milking, cleaning of the floor between groups and use of water to clean teats. 'Attention of the milkers' was also significantly associated with CMSCC (P = 0.012) and YASCC (P = 0.014). It included information on the accuracy of mastitis detection by foremilk screening and the regular use of post-milking teat and cluster disinfection. The component 'preparation routines' (method of udder cleaning and forestripping) did not significantly influence CMSCC and YASCC. These results indicate that excessive use of water in the parlour during milking time is harmful to udder health and that the consistency of procedures in the milking parlour presents significant room for improvement in large dairy herds in Brandenburg. PMID:16629957

  8. The effect of bluetongue virus serotype 8 on milk production and somatic cell count in Dutch dairy cows in 2008.

    PubMed

    Santman-Berends, I M G A; Hage, J J; Lam, T J G M; Sampimon, O C; van Schaik, G

    2011-03-01

    The effect of bluetongue virus serotype 8 (BTV-8) infections was quantified on milk production and udder health. From July 2008 to December 2008, 1,074 seronegative cows in 15 herds that were not vaccinated against BTV-8 were tested every 3 wk for BTV-8 antibodies. Sampling stopped when cows seroconverted. Test-day records were provided and 3 traits were defined to evaluate the effect of BTV-8 on milk production and udder health: 1) the difference between observed and predicted fat- and protein-corrected milk production; 2) the natural logarithm of the somatic cell count (lnSCC); and 3) the occurrence of a new high SCC. In the default model, the variables were assumed influenced by BTV-8 when the test-day record of the seroconverted cow was taken within 30 d before seroconversion, thus, in the period in which the cow was infected. In sensitivity analyses, the time intervals were varied in which BTV-8 was assumed to affect milk production and udder health. During the study, 185 cows (17%) had a subclinical infection and seroconverted and 77 had a test-day result within 30 d before seroconversion. In this period, in cows that seroconverted, the fat- and protein-corrected milk production was 52 (95% confidence interval: 26 to 77) kg less than in the period before and after seroconversion and was 51 (95% CI: 26 to 76) kg less than in cows that remained seronegative. When the time interval was increased to within 42 d before seroconversion, the milk production in BTV-8-seroconverted cows decreased by 61 (95% CI: 28 to 94) kg compared with the period before and after seroconversion and decreased by 59 (95% CI: 27 to 92) kg compared with cows that remained BTV-8 seronegative. No significant effect of BTV-8 was found on SCC and odds for a high SCC. Subclinical BTV-8 infection in dairy cattle results in a decreased milk production. PMID:21338800

  9. Quantification of Clara cell protein in rat and mouse biological fluids using a sensitive immunoassay.

    PubMed

    Halatek, T; Hermans, C; Broeckaert, F; Wattiez, R; Wiedig, M; Toubeau, G; Falmagne, P; Bernard, A

    1998-03-01

    Clara cell protein is a 16-17 kDa protein (CC16) secreted by Clara cells in the bronchiolar lining fluid of the lung. In order to investigate the potential of this protein as a pulmonary marker in animals, CC16 was isolated from rat bronchoalveolar lavage fluid (BALF) and a sensitive latex immunoassay applicable to both rat and mouse CC16 was developed. The pattern of CC16 concentrations in rat biological fluids determined by the immunoassay was consistent with the hypothesis of a passive diffusion of the protein across the bronchoalveolar/blood barriers showing a difference of more than 5,000 fold between the concentration in the epithelial lining fluid (mean, 140 mg x L(-1)) and that in serum (20 microg x L(-1)) or urine (3 microg x L(-1)). In BALF, the CC16 concentration averaged 5,500 microg x L(-1) and was of the same magnitude as that determined on lung and trachea homogenates. CC16 was also detectable in amniotic fluid with a mean value of 800 microg x L(-1) before delivery. Damage of Clara cells produced by methylcyclopentadienyl manganese tricarbonyl resulted in a significant decrease of CC16 in BALF but did not affect the serum levels of the protein. The nephrotoxicant sodium chromate by contrast had no influence on the CC16 content of BALF but markedly increased CC16 levels in both serum and urine as a result of impaired glomerular filtration and tubular reabsorption, respectively. In conclusion, mouse or rat Clara cell protein of 16-17 kDa can easily be quantified, not only in bronchoalveolar lavage fluid, but also in extrapulmonary fluids such as serum or urine. Thus, in rodents, Clara cell protein of 16-17 kDa follows the same metabolic pathway as in humans, diffusing from the respiratory tract into serum where it is eliminated by the kidneys. This serum Clara cell protein of 16-17 kDa may be useful as a peripheral marker of events taking place in the respiratory tract. PMID:9596129

  10. Androgen action via testicular arteriole smooth muscle cells is important for Leydig cell function, vasomotion and testicular fluid dynamics.

    PubMed

    Welsh, Michelle; Sharpe, Richard M; Moffat, Lindsey; Atanassova, Nina; Saunders, Philippa T K; Kilter, Sigrid; Bergh, Anders; Smith, Lee B

    2010-01-01

    Regulation of blood flow through the testicular microvasculature by vasomotion is thought to be important for normal testis function as it regulates interstitial fluid (IF) dynamics which is an important intra-testicular transport medium. Androgens control vasomotion, but how they exert these effects remains unclear. One possibility is by signalling via androgen receptors (AR) expressed in testicular arteriole smooth muscle cells. To investigate this and determine the overall importance of this mechanism in testis function, we generated a blood vessel smooth muscle cell-specific AR knockout mouse (SMARKO). Gross reproductive development was normal in SMARKO mice but testis weight was reduced in adulthood compared to control littermates; this reduction was not due to any changes in germ cell volume or to deficits in testosterone, LH or FSH concentrations and did not cause infertility. However, seminiferous tubule lumen volume was reduced in adult SMARKO males while interstitial volume was increased, perhaps indicating altered fluid dynamics; this was associated with compensated Leydig cell failure. Vasomotion was impaired in adult SMARKO males, though overall testis blood flow was normal and there was an increase in the overall blood vessel volume per testis in adult SMARKOs. In conclusion, these results indicate that ablating arteriole smooth muscle AR does not grossly alter spermatogenesis or affect male fertility but does subtly impair Leydig cell function and testicular fluid exchange, possibly by locally regulating microvascular blood flow within the testis. PMID:21049031

  11. Counting Money

    NSDL National Science Digital Library

    areese

    2008-09-25

    Today you are going to practice counting money. We will be reviewing the penny, nickel, and dime, and quarter. The coin with the lowest value is the penny. Here is a picture of a penny. A penny is worth one cent or $0.01picture of a penny The next coin of the lowest value is the nickel. Here is a picture of a nickel. picture of a nickel A nickel is worth five cents or $0.05 The next coin ...

  12. Relationships of milk culture status at calving with somatic cell counts and milk production of dairy heifers during early lactation on a Californian dairy

    Microsoft Academic Search

    John H. Kirk; Jim C. Wright; Steven L. Berry; James P. Reynolds; John P. Maas; Abbas Ahmadi

    1996-01-01

    Four-quarter, composite milk samples were collected from 339 heifers calving for the first time in a large Californian dairy which consistently had low herd somatic cell counts and low prevalence of major mastitis pathogens. The milk samples were collected on average at 6.4 days post partum (range 1–17). Thirty-nine percent of the heifers were subclinically infected with coagulase-negative Staphylococcus spp.

  13. Variability of Test-Day Milk Production and Composition and Relation of Somatic Cell Counts with Yield and Compositional Changes of Bovine Milk

    Microsoft Academic Search

    K. F. Ng-Kwai-Hang; J. F. Hayes; J. E. Moxley; H. G. Monardes

    1984-01-01

    Between November 1979 and No- vember 1981, 41,783 test-day observa- tions were obtained from 63 Holstein herds in the province of Quebec. Measured were milk yield, percentages of fat, protein, casein, and serum protein, and somatic cell counts that had unadjusted means with standard errors 20.44 + .04 kg, 3.684 + .003%, 3.314 + 002%, 2.694 + .001, .699 -+

  14. A rapid effect of handling on counts of white blood cells in a wintering passerine bird: a more practical measure of stress?

    Microsoft Academic Search

    Dina C?rule; Tatjana Krama; Jolanta Vrublevska; Markus J. Rantala; Indrikis Krams

    Measuring circulating glucocorticoids is a widely used method to assess stress in animals. However, hormones must be sampled\\u000a within the first few minutes of capture, which makes it difficult to discriminate between hormone baseline levels and the\\u000a levels caused by capture and handling stress. The use of white blood cell (WBC) counts made from blood smears represents an\\u000a alternate method

  15. The economic impact of mastitis-control procedures used in Scottish dairy herds with high bulk-tank somatic-cell counts

    Microsoft Academic Search

    C Yalcin; A. W Stott; D. N Logue; J Gunn

    1999-01-01

    We used multiple-regression analysis of field data to quantify the marginal impacts of various mastitis-control procedures on bulk-tank somatic-cell count (BTSCC). Estimates of milk-yield depression and the probability of herds paying a BTSCC penalty due to the presence of subclinical mastitis were made. An assessment of the economic efficiency of mastitis control by high BTSCC producers was also made using

  16. Atypical manifestation of progressive outer retinal necrosis in AIDS patient with CD4+ T-cell counts more than 100 cells/microL on highly active antiretroviral therapy.

    PubMed

    Vichitvejpaisal, Pornpattana; Reeponmahar, Somporn; Tantisiriwat, Woraphot

    2009-06-01

    Typical progressive outer retinal necrosis (PORN) is an acute ocular infectious disease in acquired immunodeficiency syndrome (AIDS) patients with extremely low CD4+ T-cell counts. It is a form of the Varicella- zoster virus (VZV) infection. This destructive infection has an extremely rapid course that may lead to blindness in affected eyes within days or weeks. Attempts at its treatment have had limited success. We describe the case of a bilateral PORN in an AIDS patient with an initial CD4+ T-cell count >100 cells/microL that developed after initiation of highly active antiretroviral therapy (HAART). A 29-year-old Thai female initially diagnosed with human immunodeficiency virus (HIV) in 1998, presented with bilaterally decreased visual acuity after initiating HAART two months earlier. Multiple yellowish spots appeared in the deep retina without evidence of intraocular inflammation or retinal vasculitis. Her CD4+ T-cell count was 127 cells/microL. She was diagnosed as having PORN based on clinical features and positive VZV in the aqueous humor and vitreous by polymerase chain reaction (PCR). Despite combined treatment with intravenous acyclovir and intravitreous ganciclovir, the patient's visual acuity worsened with no light-perception in either eye. This case suggests that PORN should be included in the differential diagnosis of reduced visual acuity in AIDS patients initiating HAART with higher CD4+ T-cell counts. PORN may be a manifestation of the immune reconstitution syndrome. PMID:19702067

  17. Cytologic features of ovarian granulosa cell tumors in pleural and ascitic fluids.

    PubMed

    Omori, Makiko; Kondo, Tetsuo; Yuminamochi, Tsutomu; Nakazawa, Kumiko; Ishii, Yoshio; Fukasawa, Hiroko; Hashi, Akihiko; Hirata, Shuji

    2015-07-01

    Adult granulosa cell tumor (AGCT) is an uncommon neoplasm of the ovary with potential for aggressive behavior and late recurrence. The most important prognostic factor for AGCT is tumor stage. Thus, cytological assessment of pleural or ascitic fluids is crucial for initial staging and subsequent patient management. We report herein two cases of ovarian AGCT presenting with exfoliated tumor cells in pleural and ascitic fluid. The first case involved a 61-year-old woman who presented with stage Ic (a) AGCT. Seven years after initial diagnosis, pleural effusion and pleural dissemination were identified. The second case involved a 50-year-old woman who presented with stage IV AGCT with massive ascites and right pleural effusion. Fluid cytology from both cases showed cohesive or loose clusters of small uniform neoplastic cells with round-to-oval nuclei, coffee-bean-shaped nuclear grooves, small nucleoli, and scant cytoplasm. Call-Exner bodies were also observed in these cytologic specimens. In the differential diagnosis of small monomorphic tumor cells in pleural effusion or ascites, coffee-bean-shaped nuclear grooves and cell clusters forming Call-Exner bodies are diagnostic clues of AGCT. Diagn. Cytopathol. 2015;43:581-584. © 2015 Wiley Periodicals, Inc. PMID:25605680

  18. Self-similar relaxation dynamics of a fluid wedge in a Hele-Shaw cell

    NASA Astrophysics Data System (ADS)

    Gat, Omri; Meerson, Baruch; Vilenkin, Arkady

    2006-06-01

    Let the interface between two immiscible fluids in a Hele-Shaw cell have, at t=0 , a wedge shape. As a wedge is scale-free, the fluid relaxation dynamics are self-similar. We find the dynamic exponent of this self-similar flow and show that the interface shape is given by the solution of an unusual inverse problem of potential theory. We solve this problem analytically for an almost flat wedge, and numerically otherwise. The wedge solution is useful for analysis of pinch-off singularities.

  19. The effects of HIV-1 subtype and ethnicity on the rate of CD4 cell count decline in patients naive to antiretroviral therapy: a Canadian?European collaborative retrospective cohort study

    PubMed Central

    Young, Jim; Dunn, David; Ledergerber, Bruno; Sabin, Caroline; Cozzi-Lepri, Alessandro; Dabis, Francois; Harrigan, Richard; Tan, Darrell H.; Walmsley, Sharon; Gill, John; Cooper, Curtis; Scherrer, Alexandra U.; Mocroft, Amanda; Hogg, Robert S.; Smaill, Fiona

    2014-01-01

    Background Ethnic differences have the potential to confound associations between HIV-1 subtype and immunologic progression. We compared declines in CD4 cell counts during untreated infection for the most prevalent HIV-1 subtypes, focusing on distinguishing between the effects of viral subtype and ethnicity. Methods We combined data from 4 European and 6 Canadian cohorts, selecting adults in the stable chronic phase of untreated HIV infection. We estimated the change in square root CD4 cell count over time for subtypes and ethnicities using mixed models, adjusting for covariates selected for their potential effect on initial CD4 cell count or its decline. Results Data from 9772 patients were analyzed, contributing 79 175 measurements of CD4 cell count and 24 157 person-years of follow-up. Overall, there were no appreciable differences in CD4 cell count decline for viral subtypes A, CRF01_AE, CRF02_AG, C and G compared with viral subtype B; whereas the decline in CD4 cell count in patients of African ancestry was considerably slower than in patients of other ethnicity. When ethnic groups were studied separately, there was evidence for slower declines in CD4 cell count in viral subtypes C, and possibly A and G, compared with viral subtype B in patients of African ancestry but not among patients of other ethnicities, suggesting an interaction between subtype and ethnicity. Interpretation Ethnicity is a major determinant of CD4 cell count decline; viral subtype differences may have existed but were small compared with the effect of ethnicity and were most apparent in patients of African ancestry. In developing countries, slower CD4 cell count declines among individuals of African descent may translate to a longer asymptomatic phase and increase the opportunity for HIV transmission. PMID:25485259

  20. A Higher Correlation of HCV Core Antigen with CD4+ T Cell Counts Compared with HCV RNA in HCV/HIV-1 Coinfected Patients

    PubMed Central

    Zhang, Weidong; Xi, Yuanlin; Cao, Guanghua; Zhi, Yuhong; Wang, Shuiwang; Xu, Chunhui; Wei, Lai; Lu, Fengmin; Zhuang, Hui

    2011-01-01

    Development of HCV infection is typically followed by chronic hepatitis C (CHC) in most patients, while spontaneous HCV viral clearance (SVC) occurs in only a minority of subjects. Compared with the widespread application of HCV RNA testing by quantitative RT-PCR technique, HCV core antigen detection may be an alternative indicator in the diagnosis of hepatitis C virus infections and in monitoring the status of infectious individuals. However, the correlation and differences between these two indicators in HCV infection need more investigation, especially in patients coinfected by HIV-1. In this study, a total of 354 anti-HCV and/or anti-HIV serum positive residents from a village of central China were enrolled. Besides HCV-related hepatopathic variables including clinical status, ALT, AST, anti-HCV Abs, as well as the altered CD4+/CD8+ T cell counts, HCV core antigen and HCV viral load were also measured. The concentration of serum HCV core antigen was highly correlated with level of HCV RNA in CHC patients with or without HIV-1 coinfection. Of note, HCV core antigen concentration was negatively correlated with CD4+ T cell count, while no correlation was found between HCV RNA level and CD4+ T cell count. Our findings suggested that quantitative detection of plasma HCV core antigen may be an alternative indicator of HCV RNA qPCR assay when evaluating the association between HCV replication and host immune status in HCV/HIV-1 coinfected patients. PMID:21858166

  1. Neuronal death and synapse elimination in the olivocerebellar system. II. Cell counts in the inferior olive of adult x-irradiated rats and weaver and reeler mutant mice

    SciTech Connect

    Shojaeian, H.; Delhaye-Bouchaud, N.; Mariani, J.

    1985-02-15

    Cell death in the developing rat inferior olive precedes the regression of the polyneuronal innervation of Purkinje cells by olivary axons (i.e., climbing fibers), suggesting that the involution of the redundant olivocerebellar contacts is caused by a withdrawal of supernumerary axonal collaterals rather than by degeneration of the parent cell. However, a subsequent apparent increase of the olivary population occurs, which could eventually mask a residual presynaptic cell death taking place at the same time. Therefore, cell counts were performed in the inferior olive of adult rodents in which the multiple innervation of Purkinje cells by olivary axons is maintained, with the idea that if cell death plays a role in the regression of supernumerary climbing fibers, the number of olivary cells should be higher in these animals than in their controls. The results show that the size of the cell population in the inferior olive of weaver and reeler mutant mice and rats degranulated by early postnatal x-irradiation does not differ significantly from that of their controls. Similarly, the distribution of the cells in the four main olivary subnuclei is not modified in weaver mice and x-irradiated rats. The present data further support the assumption that the regression of the polyneuronal innervation of Purkinje cells occurs independently of cell death in the presynaptic population.

  2. Cerebrospinal fluid B cells from Multiple Sclerosis patients are subject to normal germinal center selection

    PubMed Central

    Harp, Christopher; Lee, Jane; Lambracht-Washington, Doris; Cameron, Elizabeth; Olsen, Gregory; Frohman, Elliot; Racke, Michael; Monson, Nancy

    2007-01-01

    Previous findings from our laboratory demonstrated that some clonally expanded cerebrospinal fluid (CSF) B cells from MS patients exhibit diminished mutation targeting patterns in comparison to typical B cells selected in the context of germinal centers (GCs). In order to determine whether the overall CSF B cell repertoires adhered to mutation patterns typical of GC-selected B cells, we analyzed the immunoglobulin repertoires from CSF B cells of 8 MS patients for mutation characteristics typical of GC-derived B cells. Mutation targeting was preserved. Thus, clonal expansion of some CSF B cells may occur independently of GC, but the CSF B cell pool is governed by typical GC selection. Interestingly, the heavy chain CDR3’s of CSF B cells from MS patients had a net acidic charge, similar to GC-derived B cells, but a tendency towards longer CDR3’s, consistent with autoreactive B cells. How these findings may support current hypotheses regarding the origin of CSF B cells is discussed. PMID:17169437

  3. The interaction between a solid body and viscous fluid by marker-and-cell method

    NASA Technical Reports Server (NTRS)

    Cheng, R. Y. K.

    1976-01-01

    A computational method for solving nonlinear problems relating to impact and penetration of a rigid body into a fluid type medium is presented. The numerical techniques, based on the Marker-and-Cell method, gives the pressure and velocity of the flow field. An important feature in this method is that the force and displacement of the rigid body interacting with the fluid during the impact and sinking phases are evaluated from the boundary stresses imposed by the fluid on the rigid body. A sample problem of low velocity penetration of a rigid block into still water is solved by this method. The computed time histories of the acceleration, pressure, and displacement of the block show food agreement with experimental measurements. A sample problem of high velocity impact of a rigid block into soft clay is also presented.

  4. An altered repertoire of T cell receptor V gene expression by rheumatoid synovial fluid T lymphocytes.

    PubMed Central

    Lunardi, C; Marguerie, C; So, A K

    1992-01-01

    The pattern of T cell receptor V gene expression by lymphocytes from rheumatoid synovial fluid and paired peripheral blood samples was compared using a polymerase chain reaction (PCR)-based assay. Eight rheumatoid arthritis (RA) patients who had varying durations of disease (from 2 to 20 years) were studied. In all patients there was evidence of a different pattern of V gene expression between the two compartments. Significantly increased expression of at least one V alpha or V beta gene family by synovial fluid T cells was observed in all the patients studied. Three different V alpha (V alpha 10, 15 and 18) and three V beta (V beta 4, 5 and 13) families were commonly elevated. Sequencing of synovial V beta transcripts demonstrated that the basis of increased expression of selected V gene families in the synovial fluid was due to the presence of dominant clonotypes within those families, which constituted up to 53% of the sequences isolated from one particular synovial V gene family. There were considerable differences in the NDJ sequences found in synovial and peripheral blood T cell receptor (TCR) transcripts of the same V beta gene family. These data suggest that the TCR repertoire in the two compartments differs, and that antigen-driven expansion of particular synovial T cell populations is a component of rheumatoid synovitis, and is present in all stages of the disease. PMID:1458680

  5. Biodiversity Counts

    NSDL National Science Digital Library

    1998-01-01

    This extensive collection of activities from the American Museum of Natural History offers middle school students an exciting and creative context for involving students in the scientific process while introducing them to the rich diversity and beauty of their local ecosystem. Lesson plans, Web-based interactive activities, useful Web links, profiles of AMNH scientists and staff, and other features help students inventory and analyze the plants and arthropods found in their own neighborhoods. All activities address national science standards, and have been field tested in schools around the nation. Biodiversity Counts even has students develop their own exhibitions for their findings -- a great way to build science communication skills.

  6. Bone cell mechanosensation of fluid flow stimulation: a fluid-structure interaction model characterising the role integrin attachments and primary cilia.

    PubMed

    Vaughan, T J; Mullen, C A; Verbruggen, S W; McNamara, L M

    2015-08-01

    Load-induced fluid flow acts as an important biophysical signal for bone cell mechanotransduction in vivo, where the mechanical environment is thought to be monitored by integrin and primary cilia mechanoreceptors on the cell body. However, precisely how integrin- and primary cilia-based mechanosensors interact with the surrounding fluid flow stimulus and ultimately contribute to the biochemical response of bone cells within either the in vitro or in vivo environment remains poorly understood. In this study, we developed fluid-structure interaction models to characterise the deformation of integrin- and primary cilia-based mechanosensors in bone cells under fluid flow stimulation. Under in vitro fluid flow stimulation, these models predicted that integrin attachments on the cell-substrate interface were highly stimulated [Formula: see text], while the presence of a primary cilium on the cell also resulted in significant strain amplifications, arising at the ciliary base. As such, these mechanosensors likely play a role in mediating bone mechanotransduction in vitro. Under in vivo fluid flow stimulation, integrin attachments along the canalicular wall were highly stimulated and likely play a role in mediating cellular responses in vivo. The role of the primary cilium as a flow sensor in vivo depended upon its configuration within the lacunar cavity. Specifically, our results showed that a short free-standing primary cilium could not effectively fulfil a flow sensing role in vivo. However, a primary cilium that discretely attaches the lacunar wall can be highly stimulated, due to hydrodynamic pressure in the lacunocanalicular system and, as such, could play a role in mediating bone mechanotransduction in vivo. PMID:25399300

  7. A Study of Alternate Biomarkers in HIV Disease and Evaluating their Efficacy in Predicting T CD4+ Cell Counts and Disease Progression in Resource Poor Settings in Highly Active Antiretroviral Therapy (HAART) Era

    PubMed Central

    Ramana, K V; Sabitha, V; Rao, Ratna

    2013-01-01

    Introduction: Human Immunodeficiency Virus (HIV), the causative agent of AIDS, has been a challenge to medical fraternity since it was first discovered in 1983. About 40 million people are living with HIV infection globally and 99% of the infected people are in south East Asia (SEA). Traditionally, HIV disease and progression, initiation of HAART and response to therapy is monitored by assessing in regular intervals, the T CD4+ cell counts and plasma HIV/RNA viral load. Resource poor, low and low – middle income group countries still have no finances to acquire infrastructure and scientific technology for performing such tests. Objectives: Since very few studies are available, they have demonstrated the role of alternate biomarkers that can be used to predict CD4 cell counts and thereby, monitor HIV disease progression and HAART. We aimed to measure certain haematological parameters in HIV seropositive patients and to evaluate their efficacy in predicting TCD4+ cell counts. Methods: The study group included 250 HIV seropositive patients with an age range of 18-65 years. 140(56%) males and 110(44%) females were included in the study. Absolute TCD4+cell counts and CD8+T cell counts were measured by using a flow cytometer. (MMWR Recommendations and Reports, 1992) TLC; HB%, AEC and ESR were estimated by using conventional haematological methods. CRP was evaluated by latex agglutination test (Immuno CRP Latex Agglutination Test). Results: Among the tested haematological markers, a TLC of <1800 cells/mm3 showed high specificity (100%) in predicting CD4 counts of < 200 cells/mm3, with an accuracy of 61.46%. Haemoglobin and Absolute Eosinophilic counts showed high specificities of 84.09% and 94.32% respectively in predicting CD4 counts which were below 350 cells/mm3. ESR with 98.98% sensitivity and AEC which had 83.67% sensitivity were able to predict CD4 counts of <200 cells/mm3. Conclusion: Among the tested biomarkers, it was seen that Absolute Eosinophilic counts of more than 550 cells/mm3, Blood Haemoglobin which was less than 10 g%, ESR which measured more than 20 mm, CRP values of >1.2 and TLC of <1800 cells/mm3 could be helpful in predicting CD4 cell counts of < 350 and <200 cells/mm3. PMID:23998059

  8. Hematopoietic progenitor cell collection after autologous transplant for multiple myeloma: low platelet count predicts for poor collection and sole use of resulting graft enhances risk of myelodysplasia.

    PubMed

    Papanikolaou, X; Rosenbaum, E R; Tyler, L N; Sawyer, J; Heuck, C J; Barlogie, B; Cottler-Fox, M

    2014-04-01

    Collection of hematopoietic progenitor cells (HPC) after previous autologous hematopoietic progenitor cell transplant (aHCT) was studied in 221 patients with multiple myeloma (MM). With a total of 333 collections, the median number of CD34+ cells collected was 4.7 × 10(6) CD34+ cells/kg, and 74% of the patients collected ? 2.5 × 10(6) CD34+ cells/kg. Among 26 variables examined, the strongest predictor for poor collection was a platelet count <100 × 10(6)/l before mobilization (P<0.001). A subsequent aHCT was performed in 154 of the 221 patients. Sole use of HPC procured after aHCT in 86 patients was associated with delayed platelet recovery (P<0.001) and linked to development of myelodysplastic syndrome (MDS)-associated cytogenetic abnormalities (MDS-CA; P=0.027, odds ratio (OR) 10.34) and a tendency towards clinical MDS/acute myeloid leukemia (AML; P=0.091, OR 3.57). However, treatment-related mortality (P=0.766) and time to absolute neutrophil count recovery ?0.5 × 10(9)/l (P=0.879) were similar to when a pre-aHCT graft was used. Indeed, adding HPC collected before any aHCT neutralized the risk of MDS-CA or MDS/AML. Therefore, we advise generous initial HPC collection to broaden the salvage armamentarium for patients with MM. PMID:23852547

  9. Hematopoietic progenitor cell collection after autologous transplant for multiple myeloma: low platelet count predicts for poor collection and sole use of resulting graft enhances risk of myelodysplasia

    PubMed Central

    Papanikolaou, X; Rosenbaum, ER; Tyler, LN; Sawyer, J; Heuck, CJ; Barlogie, B; Cottler-Fox, M

    2014-01-01

    Collection of hematopoietic progenitor cells (HPC) after previous autologous hematopoietic progenitor cell transplant (aHCT) was studied in 221 patients with multiple myeloma (MM). With a total of 333 collections, the median number of CD34 cells collected was 4.7 × 106 CD34 + cells/kg, and 74% of the patients collected ?2.5 × 106 CD34 + cells/kg. Among 26 variables examined, the strongest predictor for poor collection was a platelet count <100 × 106/l before mobilization (P<0.001). A subsequent aHCT was performed in 154 of the 221 patients. Sole use of HPC procured after aHCT in 86 patients was associated with delayed platelet recovery (P<0.001) and linked to development of myelodysplastic syndrome (MDS)-associated cytogenetic abnormalities (MDS-CA; P = 0.027, odds ratio (OR) 10.34) and a tendency towards clinical MDS/acute myeloid leukemia (AML; P = 0.091, OR 3.57). However, treatment-related mortality (P = 0.766) and time to absolute neutrophil count recovery ?0.5 × 109/l (P = 0.879) were similar to when a pre-aHCT graft was used. Indeed, adding HPC collected before any aHCT neutralized the risk of MDS-CA or MDS/AML. Therefore, we advise generous initial HPC collection to broaden the salvage armamentarium for patients with MM. PMID:23852547

  10. Hydrothermal diamond-anvil cell: Application to studies of geologic fluids

    USGS Publications Warehouse

    Chou, I.-Ming

    2003-01-01

    The hydrothermal diamond-anvil cell (HDAC) was designed to simulate the geologic conditions of crustal processes in the presence of water or other fluids. The HDAC has been used to apply external pressure to both synthetic and natural fluid inclusions in quartz to minimize problems caused by stretching or decrepitation of inclusions during microthermometric analysis. When the HDAC is loaded with a fluid sample, it can be considered as a large synthetic fluid inclusion and therefore, can be used to study the PVTX properties as well as phase relations of the sample fluid. Because the HDAC has a wide measurement pressure-temperature range and also allows in-situ optical observations, it has been used to study critical phenomena of various chemical systems, such as the geologically important hydrous silicate melts. It is possible, when the HDAC is combined with synchrotron X-ray sources, to obtain basic information on speciation and structure of metal including rare-earth elements (REE) complexes in hydrothermal solutions as revealed by X-ray absorption fine structure (XAFS) spectra. Recent modifications of the HDAC minimize the loss of intensity of X-rays due to scattering and absorption by the diamonds. These modifications are especially important for studying elements with absorption edges below 10 keV and therefore particularly valuable for our understanding of transport and deposition of first-row transition elements and REE in hydrothermal environments.

  11. Different screening tests and milk somatic cell count for the prevalence of subclinical bovine mastitis in Bangladesh.

    PubMed

    Hoque, Md Nazmul; Das, Ziban Chandra; Talukder, Anup Kumar; Alam, Mohammad Shah; Rahman, Abu Nasar Md Aminoor

    2015-01-01

    Identification of cows with subclinical mastitis (SCM) is an important tool for sustainable dairying and implementing effective mastitis control strategies. A total of 892 quarters milk samples from 228 lactating cows were screened by California mastitis test (CMT), White side test (WST), Surf field mastitis test (SFMT), and somatic cell count (SCC) to study the prevalence of bovine SCM in some selected areas of Bangladesh. Out of 228 cows, 148 (64.9%), 138 (60.5%), 132 (57.9%), and 164 (71.9%) were found positive for SCM by CMT, WST, SFMT, and SCC, respectively. The prevalence of bovine SCM was diagnosed 45.7, 40.2, 36.6, and 29.6% in Chittagong, Sirajgonj, Mymensingh, and Gazipur districts, respectively, based on a combination of all tests. The overall quarter-wise prevalence of SCM was 45.7, 43.5, 41.2, and 55.0% for CMT, WST, SFMT, and SCC. Single quarters and left front quarters were more prone to SCM (P?CMT>WST>SFMT). Thus, CMT was concluded to be the most accurate (r?=?0.782) field diagnostic test after laboratory test like SCC (r?=?0.924). However, the use of any single test may not be reliable in diagnosing SCM, while the result of CMT supported by SCC might be used effectively to pinpoint diagnosis of SCM in dairy animals than alone. PMID:25326717

  12. Joint estimation of genetic parameters for test-day somatic cell count and mastitis in the United Kingdom.

    PubMed

    Mrode, R; Pritchard, T; Coffey, M; Wall, E

    2012-08-01

    Genetic parameters were estimated in a joint analysis of log(e)-transformed somatic cell count (TSCC) with either mastitis as a binary trait (MAS) or the number of mastitis cases (NMAS) in Holstein-Friesian cows for the first 3 lactations using a random regression model. In addition, a multi-trait analysis of MAS and NMAS was also implemented. There were 67,175, 30,617, and 16,366 cows with records for TSCC, MAS, and NMAS in lactations 1, 2, and 3, respectively. The frequency of MAS was 14, 20, and 25% in lactations 1, 2, and 3 respectively. The model for TSCC included herd-test-day, age at calving and month of calving, fixed lactation curves nested with calving year groups, and random regressions with Legendre polynomials of order 2 for animal and permanent environmental effects. The model for MAS and NMAS included fixed herd-year-season, age at calving and month of calving, and random animal and permanent environmental effects. All analyses were carried out using Gibbs sampling. Estimates of mean daily heritability averaged over a 305-d lactation were 0.11, 0.14, and 0.15 for TSCC for lactations 1, 2, and 3, respectively. Corresponding heritability estimates for MAS were 0.05, 0.07, and 0.09. The heritabilities for NMAS were similar at 0.06, 0.07, and 0.12, respectively, for lactations 1, 2, and 3. The genetic correlations between lactations 1 and 2, 1 and 3, and 2 and 3 were 0.75, 0.64, and 0.92 for computed 305-d lactation TSCC; 0.55, 0.48, and 0.89 for MAS; and 0.62, 0.42, and 0.85 for NMAS, respectively. The genetic correlations between MAS and TSCC were positive and generally moderate to high. The genetic correlations between computed 305-d lactation TSCC and MAS were 0.53, 0.61, and 0.68 in lactations 1, 2, and 3, respectively. Similar corresponding genetic correlations were obtained between computed 305-d lactation TSCC and NMAS in the respective parities. Mastitis as a binary trait and NMAS in the same lactation were very highly correlated and were genetically the same trait. It is intended that the new parameters will be used in setting up a national evaluation system for the joint analysis of TSCC and MAS. PMID:22818477

  13. ‘Living cantilever arrays’ for characterization of mass of single live cells in fluids

    PubMed Central

    Park, Kidong; Jang, Jaesung; Irimia, Daniel; Sturgis, Jennifer; Lee, James; Robinson, J. Paul; Toner, Mehmet; Bashir, Rashid

    2013-01-01

    The size of a cell is a fundamental physiological property and is closely regulated by various environmental and genetic factors. Optical or confocal microscopy can be used to measure the dimensions of adherent cells, and Coulter counter or flow cytometry (forward scattering light intensity) can be used to estimate the volume of single cells in a flow. Although these methods could be used to obtain the mass of single live cells, no method suitable for directly measuring the mass of single adherent cells without detaching them from the surface is currently available. We report the design, fabrication, and testing of ‘living cantilever arrays’, an approach to measure the mass of single adherent live cells in fluid using silicon cantilever mass sensor. HeLa cells were injected into microfluidic channels with a linear array of functionalized silicon cantilevers and the cells were subsequently captured on the cantilevers with positive dielectrophoresis. The captured cells were then cultured on the cantilevers in a microfluidic environment and the resonant frequencies of the cantilevers were measured. The mass of a single HeLa cell was extracted from the resonance frequency shift of the cantilever and was found to be close to the mass value calculated from the cell density from the literature and the cell volume obtained from confocal microscopy. This approach can provide a new method for mass measurement of a single adherent cell in its physiological condition in a non-invasive manner, as well as optical observations of the same cell. We believe this technology would be very valuable for single cell time-course studies of adherent live cells. PMID:18584076

  14. Optical imaging of Ca2+-evoked fluid secretion by murine nasal submucosal gland serous acinar cells.

    PubMed

    Lee, Robert J; Limberis, Maria P; Hennessy, Michael F; Wilson, James M; Foskett, J Kevin

    2007-08-01

    Airway submucosal glands are sites of high expression of the cystic fibrosis transmembrane conductance regulator (CFTR) Cl(-) channel and contribute to fluid homeostasis in the lung. However, the molecular mechanisms of gland ion and fluid transport are poorly defined. Here, submucosal gland serous acinar cells were isolated from murine airway, identified by immunofluorescence and gene expression profiling, and used in physiological studies. Stimulation of isolated acinar cells with carbachol (CCh), histamine or ATP was associated with marked decreases in cell volume (20 +/- 2% within 62 +/- 5 s) that were tightly correlated with increases in cytoplasmic Ca(2+) concentration ([Ca(2+)](i)) as revealed by simultaneous DIC and fluorescent indicator dye microscopy. Simultaneous imaging of cell volume and the Cl(-)-sensitive fluorophore SPQ indicated that the 20% shrinkage was associated with a fall of [Cl(-)](i) from 65 mm to 28 mm, reflecting loss of 67% of cell Cl(-) content, accompanied by parallel efflux of K(+). Upon agonist removal, [Ca(2+)](i) relaxed and the cells swelled back to resting volume via a bumetanide-sensitive Cl(-) influx pathway, likely to be NKCC1. Accordingly, agonist-induced serous acinar cell shrinkage and swelling are caused by activation of solute efflux and influx pathways, respectively, and cell volume reflects the secretory state of these cells. In contrast, elevation of cAMP failed to elicit detectible volume responses, or enhance those induced by submaximal [CCh], because the magnitude of the changes were likely to be below the threshold of detection using optical imaging. Finally, when stimulated with cholinergic or cAMP agonists, cells from mice that lacked CFTR, as well as wild-type cells treated with a CFTR inhibitor, exhibited identical rates and magnitudes of shrinkage and Cl(-) efflux compared with control cells. These results provide insights into the molecular mechanisms of salt and water secretion by lung submucosal glands, and they suggest that while murine submucosal gland fluid secretion in response to cholinergic stimulation can originate from CFTR-expressing serous acinar cells, it is not dependent upon CFTR function. PMID:17525116

  15. Short-time scale variation of phytoplankton succession in Lisbon bay (Portugal) as revealed by microscopy cell counts and HPLC pigment analysis

    NASA Astrophysics Data System (ADS)

    Silva, A.; Mendes, C. R.; Palma, S.; Brotas, V.

    2008-08-01

    The phytoplankton distribution and composition in Lisbon bay was studied, at a short time scale based on a weekly sampling, during one year (April 2004 - May 2005), using microscopic examination and pigment analysis with high-performance liquid chromatography (HPLC). This work is a contribution to the knowledge on species succession and ecology of coastal communities. The frequency of the sampling permitted monitoring peak blooming and decaying, a process which frequently occurred within 1 -2 weeks. Cell counts determined that the classes Dinophyceae, Bacillariophyceae and Prymnesiophyceae dominated the assemblages. Maxima abundances and diversity of phytoplankton were observed from spring to autumn. HPLC analysis reflected the major seasonal variations observed by the cell counts and in addition detected the presence of four small sized phytoplankton classes that were not identified by microscopy. Phytoplankton counts were essential to identify the main contributing species to total chlorophyll a. Fucoxantin, peridinin and 19'-hexanoyloxyfucoxanthin appeared as good indicators for diatoms, dinoflagellates and coccolithophores, respectively, with synchronized seasonal variations and significant positive correlations.

  16. Intracellular Calcium Changes in Rat Aortic Smooth Muscle Cells in Response to Fluid Flow

    PubMed Central

    Sharma, Ritu; Yellowley, Clare E.; Civelek, Mete; Ainslie, Kristy; Hodgson, Louis; Tarbell, John M.; Donahue, Henry J.

    2015-01-01

    Vascular smooth muscle cells (VSM) are normally exposed to transmural fluid flow shear stresses, and after vascular injury, blood flow shear stresses are imposed upon them. Since Ca2+ is a ubiquitous intracellular signaling molecule, we examined the effects of fluid flow on intracellular Ca2+ concentration in rat aortic smooth muscle cells to assess VSM responsiveness to shear stress. Cells loaded with fura 2 were exposed to steady flow shear stress levels of 0.5–10.0 dyn/cm2 in a parallel-plate flow chamber. The percentage of cells displaying a rise in cytosolic Ca2+ ion concentration ([Ca2+]i) increased in response to increasing flow, but there was no effect of flow on the ([Ca2+]i) amplitude of responding cells. Addition of Gd3+ (10 ?M) or thapsigargin (50 nM) significantly reduced the percentage of cells responding and the response amplitude, suggesting that influx of Ca2+ through ion channels and release from intracellular stores contribute to the rise in ([Ca2+]i) in response to flow. The addition of nifedipine (1 or 10 ?M) or ryanodine (10 ?M) also significantly reduced the response amplitude, further defining the role of ion channels and intracellular stores in the Ca2+ response. PMID:12051621

  17. Glutathione and GSH-dependent enzymes in bronchoalveolar lavage fluid cells in response to ozone

    SciTech Connect

    Boehme, D.S.; Hotchkiss, J.A.; Henderson, R.F. (Inhalation Toxicology Research Institute, Lovelace Biomedical and Environmental Research Institute, Albuquerque, NM (United States))

    1992-02-01

    The purpose of this study was to determine if in vivo ozone exposure results in elevations in the levels of glutathione and glutathione-dependent enzymes in cells derived from bronchoalveolar lavage fluid (BALF). Our hypothesis was that, as part of a defense mechanism against oxygen toxicity, such cells would have increased levels of glutathione (GSH) in response to an oxidant stress. Female F344/N rats were exposed to 0.8 ppm ozone, 6 hr/day, for 1, 3, or 7 days, after which cells were collected by lung lavage. The GSH and GSH-peroxidase activity per milligram of protein in the cellular fraction, both necessary for reducing cellular peroxides, were elevated after 3 days of ozone exposure. After 7 days of exposure, cellular GSH had returned to control values, but the activity of glutathione reductase, the enzyme that reduces oxidized glutathione to GSH, was increased. Extracellular GSH concentration and glutathione reductase activity in BALF were also increased after 7 days of exposure. The total glutathione equivalents (GSH and GSSG, both cellular and extracellular) in BALF increased throughout the 7-day exposure, with GSH increasing first in the cells, and then in the extracellular fluid. This study demonstrated that the glutathione anti-oxidant system of BALF cells is stimulated by exposure to ozone. This response may serve to protect cells from the toxic effects of oxidant stress.

  18. Sex based levels of C-reactive protein and white blood cell count in subjects with metabolic syndrome: Isfahan Healthy Heart Program

    PubMed Central

    Gharipour, Mojgan; Ramezani, Mohammad Arash; Sadeghi, Masuomeh; Khosravi, Alireza; Masjedi, Mohsen; Khosravi-Boroujeni, Hossein; Rafieian-kopaei, Mahmoud; Sarrafzadegan, Nizal

    2013-01-01

    Background: C-reactive protein (CRP) and white blood cell (WBC) are proinflammatory markers. They are major pathophysiological for the development of metabolic syndrome (MetS). This study aimed to address the independent associations between MetS and WBC counts and serum CRP levels and evaluation of their magnitude in relation to the MetS, based on the sex in the Iranian adults. Materials and Methods: In this cross-sectional study, subjects who met the MetS criteria, based on the Adult Treatment Panel III were selected from the Isfahan Healthy Heart Program database. A questionnaire containing the demographic data, weight, height, waist, and hip circumference of the respondents was completed for each person. Blood pressure was measured and the anthropometric measurements were done, and fasting blood samples were taken for 2 h postload plasma glucose (2 hpp). Serum [total, high-density lipoprotein (HDL), and low-density lipoprotein] levels of cholesterol, triglyceride, and CRP as well as WBC counts were determined. The univariate analyses were carried out to assess the relation between the CRP levels, WBC counts with the MetS in both sexes the. Results: In men with the abdominal obesity, the higher levels of WBC count, high serum triglyceride and blood glucose levels, a low serum HDL level, and raised systolic and diastolic blood pressure were observed. However, the higher serum CRP levels were only observed in those with the low serum HDL-cholesterol levels. The mean values of the WBC counts were statistically different between the men with and without MetS, but the mean values of the CRP levels were similar between the two groups. In women, the mean values of WBC count and CRP levels were statistically different in the subjects with and without a MetS components (except for the low serum HDL levels and high diastolic blood pressure for the WBC measures and abdominal obesity for the CRP measures) and for those with and without MetS. The age and smoking adjusted changes in the CRP levels and WBC counts correlated with the number of Mets components in the women. Conclusion: The findings of this study suggest substantial implications for the prevention and management of the MetS and atherosclerotic diseases, as these involve the suppression of inflammatory conditions rather than the incitement of anti-inflammatory conditions. PMID:24250693

  19. Endothelial cell cytoskeletal alignment independent of fluid shear stress on micropatterned surfaces

    Microsoft Academic Search

    Keri B. Vartanian; Sean J. Kirkpatrick; Stephen R. Hanson; Monica T. Hinds

    2008-01-01

    Endothelial cells (ECs) in athero-protective regions are elongated with actin and microtubule fibers aligned parallel to the direction of blood flow. Fluid shear stress (FSS) affects EC shape and functions, but little is known about shape-dependent EC properties that are independent of FSS. To evaluate these properties, ECs were elongated on micropatterned (MP) 25?m wide collagen-coated lanes (MPECs) and characterized

  20. Affordable CD4+ T cell counts by flow cytometry. II. The use of fixed whole blood in resource-poor settings.

    PubMed

    Jani, V; Janossy, G; Iqbal, A; Mhalu, F S; Lyamuya, E F; Biberfeld, G; Glencross, D K; Scott, L; Reilly, J T; Granger, V; Barnett, D

    2001-11-01

    We tested the feasibility and precision of affordable CD4+ T cell counting in resource-poor settings using a recently standardised fixative, TransFix in whole blood (WB) by flow cytometry (FCM). The precision of the assays was established under optimal conditions for single-platform FCM such as the volumetric CytoronAbsolute and the bead-based FACSCan. Fresh WB samples from HIV-seropositive and seronegative patients were tested in Tanzania and South Africa, fixed and sent to the UK for reanalysis 7 days later. Correlation, bias and limits of agreements were analysed by linear regression and the Bland-Altman test. Absolute CD4+ T cell counts remained stable for at least 10 days when TransFix was added to WB in 1:10 dilution at 20-25 degrees C, and for 7 days when added in 1:10 or 1:5 dilution to samples stored to mimic 'tropical' conditions at 37 degrees C. Higher temperatures such as 42 degrees C were tolerated for only short periods since the recovery had decreased to 63% by day 3. The reproducibility of lymphocyte subset analysis remained unchanged by TransFix with coefficient of variations <6% for all T cell subsets. Absolute CD4+ T cell counts and CD4+ T cell % values on fixed samples in the UK showed a high correlation with the results using fresh samples in Tanzania (r=0.993 and 0.969, respectively) and with the samples handled in Johannesburg (r=0.991 and 0.981) with minimal bias. Primary CD4 gating using only a single CD4 antibody also remained accurate in TransFixed samples (r=0.999). Thus, TransFix permits optimal fixation and transport of WB samples in the developing world for FCM to local regional laboratories and for quality assurance in international centres. When used together with inexpensive primary CD4 gating, TransFix will allow reliable and affordable CD4+ T cell counting by FCM in resource-poor settings. PMID:11687248

  1. Surfactant Protein D Is Present in Human Tear Fluid and the Cornea and Inhibits Epithelial Cell Invasion by Pseudomonas aeruginosa

    Microsoft Academic Search

    Minjian Ni; David J. Evans; Samuel Hawgood; E. Margot Anders; Robert A. Sack; Suzanne M. J. Fleiszig

    2005-01-01

    We have previously shown that human tear fluid protects corneal epithelial cells against Pseudomonas aeru- ginosa in vitro and in vivo and that protection does not depend upon tear bacteriostatic activity. We sought to identify the responsible tear component(s). The hypothesis tested was that collectins (collagenous calcium- dependent lectins) were involved. Reflex tear fluid was collected from healthy human subjects

  2. The quantitative effect of microextractor cell geometry on the analytical supercritical fluid extraction efficiencies of environmentally important compounds

    Microsoft Academic Search

    K. G. Furton; J. Rein

    1991-01-01

    The quantitative effect of microextractor cell geometries on supercritical fluid extraction (SFE) efficiencies of polycyclic aromatic hydrocarbons (PAHs) and methoxychlor from octadecyl-bonded sorbents has been evaluated and compared to similar effects seen upon increasing the supercritical fluid density. For the PAHs studied, correlations between the fused ring number and the relative increase in recoveries have been established. SFE recoveries can

  3. Polymer dynamics and fluid flow in actin-based cell motility

    NASA Astrophysics Data System (ADS)

    Theriot, Julie

    2005-03-01

    In living cells, nonequilibrium protein polymerization reactions are frequently used to convert chemical energy into mechanical energy and thereby generate useful force for cellular movements. We have examined the polymer and fluid dynamics in two biological cases where the assembly of branched actin filament networks generates force: the intracellular movement of the bacterial pathogen Listeria monocytogenes, and the extension of the leading edge of skin epithelial cells during wound-healing. In both cases, net actin filament assembly occurs at the front of the network structure and net disassembly occurs at the rear. Actin protein subunits and other network components must be recycled through the fluid phase to the front of the polymerizing network in order for forward movement to continue at steady state. For actin-based movement of Listeria monocytogenes, we have found that actin recycling is not rate-limiting; instead, the speed of movement is governed by the cooperative dissociation of groups of noncovalent protein-protein bonds attaching the filamentous network to the bacterial surface. In contrast, rapid actin-based extension at the leading edge of moving epithelial cells is associated with unusual perturbations in intracellular fluid flow.

  4. Mechanism of vibration-induced repulsion force on a particle in a viscous fluid cell.

    PubMed

    Saadatmand, Mehrrad; Kawaji, Masahiro

    2013-08-01

    Space platforms such as the Space Shuttle and International Space Station have been considered an ideal environment for production of protein and semiconductor crystals of superior quality due to the negligible gravity-induced convection. Although it was believed that under microgravity environment diffusive mass transport would dominate the growth of the crystals, some related experiments have not shown satisfactory results possibly due to the movement of the growing crystals in fluid cells caused by small vibrations present in the space platforms called g-jitter. In ground-based experiments, there have been clear observations of attraction and repulsion of a solid particle with respect to a nearby wall of the fluid cell due to small vibrations. The present work is a numerical investigation on the physical mechanisms responsible for the repulsion force, which has been predicted to increase with the cell vibration frequency and amplitude, as well as the fluid viscosity. Moreover, the simulations have revealed that the repulsion force occurs mostly due to the increased pressure in the narrow gap between the particle and the nearest wall. PMID:24032936

  5. Bronchoalveolar lavage fluid from normal rats stimulates DNA synthesis in rat alveolar type II cells

    SciTech Connect

    Leslie, C.C.; McCormick-Shannon, K.; Mason, R.J.

    1989-02-01

    Proliferation of alveolar type II cells after lung injury is important for the restoration of the alveolar epithelium. Bronchoalveolar lavage fluid (BALF) may represent an important source of growth factors for alveolar type II cells. To test this possibility, BALF fluid was collected from normal rats, concentrated 10-fold by Amicon filtration, and tested for its ability to stimulate DNA synthesis in rat alveolar type II cells in primary culture. BALF induced a dose-dependent increase in type II cell DNA synthesis resulting in a 6-fold increase in (3H)thymidine incorporation. Similar doses also stimulated (3H)thymidine incorporation into rat lung fibroblasts by 6- to 8-fold. Removal of pulmonary surface active material by centrifugation did not significantly reduce the stimulatory activity of BALF for type II cells. The stimulation of type II cell DNA synthesis by BALF was reduced by 100% after heating at 100 degrees C for 10 min, and by approximately 80% after reduction with dithiothreitol, and after trypsin treatment. Dialysis of BALF against 1 N acetic acid resulted in a 27% reduction in stimulatory activity. The effect of BALF in promoting type II cell DNA synthesis was more pronounced when tested in the presence of serum, although serum itself has very little effect on type II cell DNA synthesis. When BALF was tested in combination with other substances that stimulate type II cell DNA synthesis (cholera toxin, insulin, epidermal growth factor, and acidic fibroblast growth factor), additive effects or greater were observed. When BALF was chromatographed over Sephadex G150, the activity eluted with an apparent molecular weight of 100 kDa.

  6. Ocular Fluid As a Replacement for Serum in Cell Cryopreservation Media

    PubMed Central

    Venna, Naresh Kumar; Murthy, Ch Lakshmi N.; Idris, Mohammed M.; Goel, Sandeep

    2015-01-01

    Cryostorage is of immense interest in biomedical research, especially for stem cell-based therapies and fertility preservation. Several protocols have been developed for efficient cryopreservation of cells and tissues, and a combination of dimethyl sulfoxide (DMSO) and fetal bovine serum (FBS) is commonly used. However, there is a need for an alternative to FBS because of ethical reasons, high cost, and risk of contamination with blood-borne diseases. The objective of the present study was to examine the possibility of using buffalo (Bubalus bubalis) ocular fluid (BuOF) to replace FBS in cryomedia. Frozen–thawed cells, which were cryopreserved in a cryomedia with BuOF, were assessed for viability, early and late apoptosis, and proliferation. Three cell lines (CHO, HEK, and C18-4), mouse embryonic stem (mES) cells, and primary cells, such as mouse embryonic fibroblast (MEF) cells, human peripheral blood mononuclear cells (hPBMCs), and mouse bone marrow cells (mBMCs), were cryopreserved in cryomedia containing 10% DMSO (D10) with 20% FBS (D10S20) or D10 with 20% BuOF (D10O20). For all three cell lines and mES cells cryopreserved in either D10S20 or D10O20, thawed cells showed no difference in cell viability or cell recovery. Western blot analysis of frozen–thawed-cultured cells revealed that the expression of Annexin V and proliferating cell nuclear antigen (PCNA) proteins, and the ratio of BAX/BCL2 proteins were similar in all three cell lines, mES cells, and hPBMCs cryopreserved in D10S20 and D10O20. However, initial cell viability, cell recovery after culture, and PCNA expression were significantly lower in MEF cells, and the BAX/BCL2 protein ratio was elevated in mBMCs cryopreserved in D10O20. Biochemical and proteomic analysis of BuOF showed the presence of several components that may have roles in imparting the cryoprotective property of BuOF. These results encourage further research to develop an efficient serum-free cryomedia for several cell types using BuOF. PMID:26135924

  7. A Fluid Membrane-Based Soluble Ligand Display System for Live CellAssays

    SciTech Connect

    Nam, Jwa-Min; Nair, Pradeep N.; Neve, Richard M.; Gray, Joe W.; Groves, Jay T.

    2005-10-14

    Cell communication modulates numerous biological processes including proliferation, apoptosis, motility, invasion and differentiation. Correspondingly, there has been significant interest in the development of surface display strategies for the presentation of signaling molecules to living cells. This effort has primarily focused on naturally surface-bound ligands, such as extracellular matrix components and cell membranes. Soluble ligands (e.g. growth factors and cytokines) play an important role in intercellular communications, and their display in a surface-bound format would be of great utility in the design of array-based live cell assays. Recently, several cell microarray systems that display cDNA, RNAi, or small molecules in a surface array format were proven to be useful in accelerating high-throughput functional genetic studies and screening therapeutic agents. These surface display methods provide a flexible platform for the systematic, combinatorial investigation of genes and small molecules affecting cellular processes and phenotypes of interest. In an analogous sense, it would be an important advance if one could display soluble signaling ligands in a surface assay format that allows for systematic, patterned presentation of soluble ligands to live cells. Such a technique would make it possible to examine cellular phenotypes of interest in a parallel format with soluble signaling ligands as one of the display parameters. Herein we report a ligand-modified fluid supported lipid bilayer (SLB) assay system that can be used to functionally display soluble ligands to cells in situ (Figure 1A). By displaying soluble ligands on a SLB surface, both solution behavior (the ability to become locally enriched by reaction-diffusion processes) and solid behavior (the ability to control the spatial location of the ligands in an open system) could be combined. The method reported herein benefits from the naturally fluid state of the supported membrane, which allows surface-linked ligands to diffuse freely in two dimensions. Ligands can become reorganized beneath cells, by reaction-diffusion processes, and may also adopt spatial configurations reflecting those of their cognate receptors on the cell surface (Figure 1B). This provides a significant benefit over conventional cell signaling and culturing systems that present inflexible distributions of signaling molecules. In this study, we observe marked differences in the response of cells to membrane surface displayed soluble ligands as a function of membrane fluidity. Tethering of soluble signaling molecules to fluid supported membranes opens up opportunities to use already developed membrane fabrication technologies to present soluble components within a surface array format.

  8. Spindle Shaped Human Mesenchymal Stem/Stromal Cells from Amniotic Fluid Promote Neovascularization

    PubMed Central

    Pappa, Kalliopi I.; Anagnou, Nicholas P.; Watt, Suzanne M.

    2013-01-01

    Human amniotic fluid obtained at amniocentesis, when cultured, generates at least two morphologically distinct mesenchymal stem/stromal cell (MSC) subsets. Of these, the spindle shaped amniotic fluid MSCs (SS-AF-MSCs) contain multipotent cells with enhanced adipogenic, osteogenic and chondrogenic capacity. Here, we demonstrate, for the first time, the capacity of these SS-AF-MSCs to support neovascularization by umbilical cord blood (UCB) endothelial colony forming cell (ECFC) derived cells in both in vitro and in vivo models. Interestingly, although the kinetics of vascular tubule formation in vitro were similar when the supporting SS-AF-MSCs were compared with the best vasculogenic supportive batches of bone marrow MSCs (BMSCs) or human dermal fibroblasts (hDFs), SS-AF-MSCs supported vascular tubule formation in vivo more effectively than BMSCs. In NOD/SCID mice, the human vessels inosculated with murine vessels demonstrating their functionality. Proteome profiler array analyses revealed both common and distinct secretion profiles of angiogenic factors by the SS-AF-MSCs as opposed to the hDFs and BMSCs. Thus, SS-AF-MSCs, which are considered to be less mature developmentally than adult BMSCs, and intermediate between adult and embryonic stem cells in their potentiality, have the additional and very interesting potential of supporting increased neovascularisation, further enhancing their promise as vehicles for tissue repair and regeneration. PMID:23359810

  9. Spindle shaped human mesenchymal stem/stromal cells from amniotic fluid promote neovascularization.

    PubMed

    Roubelakis, Maria G; Tsaknakis, Grigorios; Pappa, Kalliopi I; Anagnou, Nicholas P; Watt, Suzanne M

    2013-01-01

    Human amniotic fluid obtained at amniocentesis, when cultured, generates at least two morphologically distinct mesenchymal stem/stromal cell (MSC) subsets. Of these, the spindle shaped amniotic fluid MSCs (SS-AF-MSCs) contain multipotent cells with enhanced adipogenic, osteogenic and chondrogenic capacity. Here, we demonstrate, for the first time, the capacity of these SS-AF-MSCs to support neovascularization by umbilical cord blood (UCB) endothelial colony forming cell (ECFC) derived cells in both in vitro and in vivo models. Interestingly, although the kinetics of vascular tubule formation in vitro were similar when the supporting SS-AF-MSCs were compared with the best vasculogenic supportive batches of bone marrow MSCs (BMSCs) or human dermal fibroblasts (hDFs), SS-AF-MSCs supported vascular tubule formation in vivo more effectively than BMSCs. In NOD/SCID mice, the human vessels inosculated with murine vessels demonstrating their functionality. Proteome profiler array analyses revealed both common and distinct secretion profiles of angiogenic factors by the SS-AF-MSCs as opposed to the hDFs and BMSCs. Thus, SS-AF-MSCs, which are considered to be less mature developmentally than adult BMSCs, and intermediate between adult and embryonic stem cells in their potentiality, have the additional and very interesting potential of supporting increased neovascularisation, further enhancing their promise as vehicles for tissue repair and regeneration. PMID:23359810

  10. Application of Fluid Mechanical Force to Embryonic Sources of Hemogenic Endothelium and Hematopoietic Stem Cells

    PubMed Central

    Li, Nan; Diaz, Miguel F.; Wenzel, Pamela L.

    2014-01-01

    During embryonic development, hemodynamic forces caused by blood flow support vascular remodeling, arterialization of luminal endothelium, and hematopoietic stem cell (HSC) emergence. Previously, we reported that fluid shear stress plays a key role in stimulating nitric oxide (NO) signaling in the aorta-gonad-mesonephros (AGM) and is essential for definitive hematopoiesis. We employed a Dynamic Flow System modified from a cone-and-plate assembly to precisely regulate in vitro exposure of AGM cells to a defined pattern of laminar shear stress. Here, we present the design of a microfluidic platform accessible to any research group that requires small cell numbers and allows for recirculation of paracrine signaling factors with minimal damage to nonadherent hematopoietic progenitors and stem cells. We detail the assembly of the microfluidic platform using commercially available components and provide specific guidance in the use of an emerging standard in the measurement of embryonic HSC potential, intravenous neonatal transplantation. PMID:25063503

  11. Neuronal death and synapse elimination in the olivocerebellar system: III. Cell counts in the inferior olive of developing rats X-irradiated from birth

    SciTech Connect

    Geoffroy, B.; Shojaeian, H.; Delhaye-Bouchaud, N.; Mariani, J.

    1988-01-08

    The change with age of cell number in the developing inferior olivary nucleus (ION) of the normal rat, compared to the time course of the regression of the polyneuronal innervation of Purkinje cells by olivary axons (i.e., the climbing fibers), suggests that the involution of the redundant olivocerebellar contacts is caused by a reduction of axonal branching rather than by degeneration of the parent cells, this being also suggested by the normal size of the olivary population in adult rodents whose Purkinje cells retain polyneuronal innervation. However, the similar size of the adult ION population does not necessarily imply that the development history is the same in normal and multiply innervated adult rodents. Therefore, cell counts were performed in developing rats which had been repeatedly X-irradiated from birth until postnatal day 14 and which retained polyneuronal innervation. The results show that, although less marked than during normal development, the evolution of the ION population is also characterized by a phase of cell loss followed by a slow increase. However, the number of cells in X-irradiated rats is higher than in their controls from birth to postnatal day 15 but becomes identical at 20 days and later. These data confirm that cell death in the ION does not play a major role in the shaping of olivocerebellar connections.

  12. Effect of Embryonic Cerebrospinal Fluid on Proliferation and Differentiation of Neuroprogenitor Cells

    PubMed Central

    Yari, Siamak; Parivar, Kazem; Nabiuni, Mohammad; Keramatipour, Mohammad

    2013-01-01

    Objective: Embryonic cerebrospinal fluid (e-CSF) has an important role in development of embryonic and adult brain. Proteomic analysis suggests that this fluid has many morphogenes and cytokines that alter in time and space throughout embryonic life. The aim of this study was to evaluate the developmental effect of embryonic CSF on proliferation and differentiation of neuroprogenitor cells in different gestational age. Materials and Methods: In this In this experimental study, we examined the role of e- CSF on proliferation and differentiation of neuroprogenitor cells using neurosphere culture method. Neurospheres size analysis and MTT assay were used to assess cell proliferation after four days in vitro. Glial differentiation study was carried out by immunocytochemistry. Neurospheres size and percentage of glial fibrialy acidic protein (GFAP) positive cells were measured by image analyzer (image J). The data were analyzed by one-way ANOVA, followed by the Tukey’s post hoc test. Data were expressed as mean ± SEM, and differences were considered significant when p<0.05, 0.01 and 0.001. Results: Viability and proliferation of neuro progenitor cells in cultures conditioned with E16 CSF and E18 CSF were significantly increased compare to control group. A dramatic decrease in percentage of GFAP-positive cells was found following the application of CSF from E16 and E18 embryos, but not E20 CSF. Conclusion: Our data suggest that, e-CSF altered proliferation and differentiation of neuro progenitor cells in age dependent manner. E16 and E18 CSF enhanced proliferation and viability of neuro progenitor cells, and inhibited differentiation to glial fate in comparison with control group. PMID:23700558

  13. New apparatus for direct counting of. beta. particles from two-dimensional gels and an application to changes in protein synthesis due to cell density

    SciTech Connect

    Anderson, H.L.; Puck, T.T.; Shera, E.B.

    1987-07-01

    A new method is described for scanning two-dimensional gels by the direct counting of ..beta.. particles instead of autoradiography. The methodology is described; results are compared with autoradiographic results; and data are presented demonstrating changed patterns of protein synthesis accompanying changes in cell density. The method is rapid and permits identification of differences in protein abundance of approximately 10% for a substantial fraction of the more prominent proteins. A modulation effect of more than 5 standard deviations, accompanying contact inhibition of cell growth, is shown to occur for an appreciable number of these proteins. The method promises to be applicable to a variety of biochemical and genetic experiments designed to delineate changes in protein synthesis accompanying changes in genome, molecular environment, history, and state of differentiation of the cell populations studied. 13 refs., 8 figs., 4 tabs.

  14. A Bereavement Support Group Intervention Is Longitudinally Associated with Salutary Effects on the CD4 Cell Count and Number of Physician Visits

    PubMed Central

    Goodkin, Karl; Feaster, Daniel J.; Asthana, Deshratn; Blaney, Nancy T.; Kumar, Mahendra; Baldewicz, Teri; Tuttle, Raymond S.; Maher, Kevin J.; Baum, Marianna K.; Shapshak, Paul; Fletcher, Mary Ann

    1998-01-01

    A randomized, controlled, clinical trial was conducted to examine the impact of a semistructured, 10-week, once weekly, 90-min/session bereavement support group intervention on immunological, neuroendocrine, and clinical health status in human immunodeficiency virus type 1-seropositive (HIV-1+) and HIV-1-seronegative (HIV-1?) homosexual men, compared to a standard of care control condition. A total of 119 homosexual men (74 HIV-1+ and 45 HIV-1?) were assessed at baseline, 10 weeks, and 6 months follow-up. At the 6-month follow-up assessment, the intervention groups exhibited significant beneficial effects compared to controls on changes in CD4 cell, total T-lymphocyte, and total lymphocyte counts, when baseline levels, antiretroviral medication use, CDC stage of disease, and other potentially confounding factors were accounted for. There was no statistically significant effect on the CD4/CD8 ratio or on the CD8 cell count. The effect on CD4 cell count was associated with group attendance and with changes in plasma cortisol level. Plasma cortisol levels decreased significantly among intervention subjects, compared to controls. A significantly reduced number of health care visits over the 6-month follow-up period among the intervention subjects supported the clinical relevance of the immunological changes observed for both HIV-1+ and HIV-1? individuals. These results indicate that behavioral interventions may have salutary immunological and clinical health effects following bereavement among HIV-1-infected individuals. The effect in HIV-1? individuals suggests that this bereavement support group intervention might have similar salutary effects in the general population. Potential effects of such interventions on clinical HIV disease progression are of interest and should be studied. PMID:9605995

  15. A bereavement support group intervention is longitudinally associated with salutary effects on the CD4 cell count and number of physician visits.

    PubMed

    Goodkin, K; Feaster, D J; Asthana, D; Blaney, N T; Kumar, M; Baldewicz, T; Tuttle, R S; Maher, K J; Baum, M K; Shapshak, P; Fletcher, M A

    1998-05-01

    A randomized, controlled, clinical trial was conducted to examine the impact of a semistructured, 10-week, once weekly, 90-min/session bereavement support group intervention on immunological, neuroendocrine, and clinical health status in human immunodeficiency virus type 1-seropositive (HIV-1+) and HIV-1-seronegative (HIV-1-) homosexual men, compared to a standard of care control condition. A total of 119 homosexual men (74 HIV-1+ and 45 HIV-1-) were assessed at baseline, 10 weeks, and 6 months follow-up. At the 6-month follow-up assessment, the intervention groups exhibited significant beneficial effects compared to controls on changes in CD4 cell, total T-lymphocyte, and total lymphocyte counts, when baseline levels, antiretroviral medication use, CDC stage of disease, and other potentially confounding factors were accounted for. There was no statistically significant effect on the CD4/CD8 ratio or on the CD8 cell count. The effect on CD4 cell count was associated with group attendance and with changes in plasma cortisol level. Plasma cortisol levels decreased significantly among intervention subjects, compared to controls. A significantly reduced number of health care visits over the 6-month follow-up period among the intervention subjects supported the clinical relevance of the immunological changes observed for both HIV-1+ and HIV-1- individuals. These results indicate that behavioral interventions may have salutary immunological and clinical health effects following bereavement among HIV-1-infected individuals. The effect in HIV-1- individuals suggests that this bereavement support group intervention might have similar salutary effects in the general population. Potential effects of such interventions on clinical HIV disease progression are of interest and should be studied. PMID:9605995

  16. A fictitious domain method with a hybrid cell model for simulating motion of cells in fluid flow

    NASA Astrophysics Data System (ADS)

    Hao, Wenrui; Xu, Zhiliang; Liu, Chun; Lin, Guang

    2015-01-01

    In this study, we develop a hybrid model to represent membranes of biological cells and use the distributed-Lagrange-multiplier/fictitious-domain (DLM/FD) formulation for simulating the fluid/cell interactions. The hybrid model representing the cellular structure consists of a continuum representation of the lipid bilayer, from which the bending force is calculated through energetic variational approach, a discrete cytoskeleton model utilizing the worm-like chain to represent network filament, and area/volume constraints. For our computational scheme, a formally second-order accurate fractional step scheme is employed to decouple the entire system into three sub-systems: a fluid problem, a solid problem and a Lagrange multiplier problem. The flow problem is solved by the projection method; the solid problem based on the cell model is solved by a combination of level set method, ENO reconstruction, and the Newton method; and the Lagrange multiplier problem is solved by immerse boundary interpolation. The incompressibility of the material is implemented with the penalty function method. Numerical results compare favorably with previously reported numerical and experimental results, and show that our method is suited to the simulation of the cell motion in flow.

  17. WBC count

    MedlinePLUS

    ... the blood is 4,500-10,000 white blood cells per microliter (mcL). Normal value ranges may vary slightly among different labs. Some laboratories use different measurements or may test different specimens. ...

  18. Calcification after Myocardial Infarction is Independent of Amniotic Fluid Stem Cell Injection

    PubMed Central

    Delo, Dawn M.; Guan, Xuan; Wang, Zhan; Groban, Leanne; Callahan, Michael; Smith, Tom; Sane, David; Payne, R. Mark; Atala, Anthony

    2010-01-01

    Ischemic heart disease remains one of the most common causes of mortality in developed countries. Recently, stem cell therapy is being considered for treating ischemic heart diseases. On the other hand, there has been evidence of chondro-osteogenic mass formation after stem cell injection in the heart. In a recent publication, Chiavegato et al. has suggested that amniotic fluid derived stem (AFS) cells cause chondro-osteogenic masses in the infarcted heart. The goal of the current study was to further examine the formation of such masses, specifically, the role of AFS cells in this process. Our results confirm the presence of similar bone-like masses in the left ventricular wall of infarcted rats; however, this phenomenon occurred independent of AFS cell injection into the myocardium. Moreover, AFS cell injection did not increase the presence of chondro-osteogenic masses. Echocardiographic analysis of large infarctions in rats frequently revealed the presence of echogenic structures in the left ventricular wall. We further demonstrated a significant relationship between the infarction size and chondro-osteogenic formation and subsequently decrease in cardiac function. Collectively, our study indicates that chondro-osteogenic differentiation can take place in infarcted rat heart independent of cell injection. These results have significant implications for future design and testing of stem cell therapy for treatment of cardiac muscle diseases. PMID:20382039

  19. Drug Transporters on Arachnoid Barrier Cells Contribute to the Blood–Cerebrospinal Fluid Barrier

    PubMed Central

    Yasuda, Kazuto; Cline, Cynthia; Vogel, Peter; Onciu, Mihaela; Fatima, Soghra; Sorrentino, Brian P.; Thirumaran, Ranjit K.; Ekins, Sean; Urade, Yoshihiro; Fujimori, Ko

    2013-01-01

    The subarachnoid space, where cerebrospinal fluid (CSF) flows over the brain and spinal cord, is lined on one side by arachnoid barrier (AB) cells that form part of the blood-CSF barrier. However, despite the fact that drugs are administered into the CSF and CSF drug concentrations are used as a surrogate for brain drug concentration following systemic drug administration, the tight-junctioned AB cells have never been examined for whether they express drug transporters that would influence CSF and central nervous system drug disposition. Hence, we characterized drug transporter expression and function in AB cells. Immunohistochemical analysis showed P-glycoprotein (P-gp) and breast cancer resistance protein (BCRP) in mouse AB cells but not other meningeal tissue. The Gene Expression Nervous System Atlas (GENSAT) database and the Allen Mouse Brain Atlas confirmed these observations. Microarray analysis of mouse and human arachnoidal tissue revealed expression of many drug transporters and some drug-metabolizing enzymes. Immortalized mouse AB cells express functional P-gp on the apical (dura-facing) membrane and BCRP on both apical and basal (CSF-facing) membranes. Thus, like blood-brain barrier cells and choroid plexus cells, AB cells highly express drug transport proteins and likely contribute to the blood-CSF drug permeation barrier. PMID:23298861

  20. Effect of amniotic fluid on the in vitro culture of human corneal endothelial cells.

    PubMed

    Feizi, Sepehr; Soheili, Zahra-Soheila; Bagheri, Abouzar; Balagholi, Sahar; Mohammadian, Azam; Rezaei-Kanavi, Mozhgan; Ahmadieh, Hamid; Samiei, Shahram; Negahban, Kambiz

    2014-05-01

    The present study was designed to evaluate the effects of human amniotic fluid (HAF) on the growth of human corneal endothelial cells (HCECs) and to establish an in vitro method for expanding HCECs. HCECs were cultured in DMEM-F12 supplemented with 20% fetal bovine serum (FBS). Confluent monolayer cultures were trypsinized and passaged using either FBS- or HAF-containing media. Cell proliferation and cell death ELISA assays were performed to determine the effect of HAF on cell growth and viability. The identity of the cells cultured in 20% HAF was determined using immunocytochemistry (ICC) and real-time reverse transcription polymerase chain reaction (RT-PCR) techniques to evaluate the expression of factors that are characteristic of HCECs, including Ki-67, Vimentin, Na+/K+-ATPase and ZO-1. HCEC primary cultures were successfully established using 20% HAF-containing medium, and these cultures demonstrated rapid cell proliferation according to the cell proliferation and death ELISA assay results. The ICC and real time RT-PCR results indicated that there was a higher expression of Na+/K+-ATPase and ZO-1 in the 20% HAF cell cultures compared with the control (20% FBS) (P < 0.05). The 20% HAF-containing medium exhibited a greater stimulatory effect on HCEC growth and could represent a potential enriched supplement for HCEC regeneration studies. PMID:24726921

  1. Lymphocyte counts and functions in arterial and venous splenic blood of patients with Hodgkin's disease. Evidence for elimination of spontaneously DNA synthesizing cells in the spleen.

    PubMed Central

    Björkholm, M; Holm, G; Askergren, J; Mellstedt, H

    1983-01-01

    Lymphocyte counts and functional competence of lymphocytes from arterial and venous splenic blood were studied in six patients with Hodgkin's disease subjected to splenectomy. One patient was untreated, four were tested after mantle field treatment and a sixth patient had a splenic relapse after total nodal radiotherapy. The percentage of E binding cells in splenic venous blood was lower than that of arterial blood though no significant differences were found in total lymphocyte or E binding cell counts. The spontaneous lymphocyte DNA synthesis was lower in venous than in arterial splenic lymphocytes in all patients. Pokeweed mitogen (PWM)-induced DNA synthesis was much lower in lymphocytes from splenic venous blood than in those from arterial blood in two patients and marginally decreased in another two. The pattern of concanavalin A response was similar to that of PWM. The elimination of lymphocytes over the spleen could not be related to the presence of lymphocytotoxic serum factors or to splenic weight or histologically verified tumour involvement. The results support the notion that some facets of the blood lymphocyte abnormalities in Hodgkin's disease may be explained by removal of functionally active lymphocyte subpopulations in the spleen. It is also concluded that spontaneously activated lymphoid cells are detained in the spleen. PMID:6872314

  2. PIMA Point of Care CD4+ Cell Count Machines in Remote MNCH Settings: Lessons Learned from Seven Districts in Zimbabwe

    PubMed Central

    Mtapuri-Zinyowera, Sekesai; Chiyaka, Edward T.; Mushayi, Wellington; Musuka, Godfrey; Naluyinda-Kitabire, Florence; Mushavi, Angella; Chikwasha, Vasco

    2013-01-01

    An evaluation was commissioned to generate evidence on the impact of PIMA point-of-care CD4+ count machines in maternal and new-born child health settings in Zimbabwe; document best practices, lessons learned, challenges, and recommendations related to scale up of this new technology. A mixed methodology approach that included 31 in-depth interviews with stakeholders involved in procurement, distribution, and use of the POC machines was employed. Additionally, data was also abstracted from 207 patient records from 35 sites with the PIMA POC CD4+ count machines and 10 other comparative sites without the machine. A clearer training strategy was found to be necessary. The average time taken to initiate clients on antiretroviral treatment (ART) was substantially less, 15 days (IQR-1-149) for sites with a PIMA POC machine as compared to 32.7 days (IQR-1-192) at sites with no PIMA POC machine. There was general satisfaction because of the presence of the PIMA POC CD4+ count machine at sites that also initiated ART. PMID:24847177

  3. Short-term Garlic Supplementation and Highly Active Antiretroviral Treatment Adherence, CD4+ Cell Counts, and Human Immunodeficiency Virus Viral Load

    PubMed Central

    Liu, Chenglong; Wang, Cuiwei; Robison, Esther; Levine, Alexandra M.; Gandhi, Monica; Schwartz, Rebecca; Weber, Kathleen M.; Merenstein, Daniel

    2012-01-01

    Context Human immunodeficiency virus (HIV)–infected individuals frequently have consumed garlic, a popular complementary supplement. Researchers rarely have studied garlic’s association with antiretroviral therapies, however, even though that association is very relevant clinically. Objective To examine associations of supplemental use of garlic with highly active antiretroviral treatment (HAART) adherence level and HAART effectiveness (HIV viral load and CD4+ cell counts) in HIV-infected women. Design The research team carried out a self-controlled, longitudinal study nested within the Women’s Interagency HIV Study (WIHS). The team used a paired study design that allowed participants to serve as their own controls. The team first identified all of the study’s visits in which the participant self-reported the use of a garlic supplement since her last visit (index visit). Then for each index visit, the team identified a matching visit (a control visit) using the following criteria: (a) the visit must be one for the same participant in which that participant reported no garlic supplementation; (b) the visit must immediately precede the index visit (less than 1 year apart); and (c) at the time of the control visit, the participant must have been using antiretroviral therapy identical to that used at the time of the index visit. Participants Participants were persons using garlic supplementation who already were participants in the WIHS. Outcome Measures The research team used a logistic regression model to examine the association between garlic supplementation and HAART adherence level. The team used a mixed linear model to examine the association of garlic supplementation with HIV viral load and CD4+ cell counts. Results From October 1994 to April 2009, 390 HIV-infected women in the WIHS made 1112 visits at which they reported using garlic supplements. Seventy-seven HIV-infected women using HAART met the research team’s selection criteria and contributed 99 pairs of visits for the study. Among the women who used garlic supplements, 22% were 50 years and older; 58% were black and non-Hispanic; and 23% had less than a high-school education. Neither use of garlic supplementation nor reasons for using garlic supplements were significantly associated with the HAART adherence level, HIV viral load, or CD4+ cell counts; however, “use garlic as needed,” a potential marker of a disease state, was significantly associated with higher viral load (P = .0003). Conclusion Short-term garlic supplementation did not impact HAART adherence level, HIV viral load, and CD4+ cell counts. PMID:22516847

  4. Fuel cell assembly unit for promoting fluid service and electrical conductivity

    DOEpatents

    Jones, Daniel O. (Glenville, NY)

    1999-01-01

    Fluid service and/or electrical conductivity for a fuel cell assembly is promoted. Open-faced flow channel(s) are formed in a flow field plate face, and extend in the flow field plate face between entry and exit fluid manifolds. A resilient gas diffusion layer is located between the flow field plate face and a membrane electrode assembly, fluidly serviced with the open-faced flow channel(s). The resilient gas diffusion layer is restrained against entering the open-faced flow channel(s) under a compressive force applied to the fuel cell assembly. In particular, a first side of a support member abuts the flow field plate face, and a second side of the support member abuts the resilient gas diffusion layer. The support member is formed with a plurality of openings extending between the first and second sides of the support member. In addition, a clamping pressure is maintained for an interface between the resilient gas diffusion layer and a portion of the membrane electrode assembly. Preferably, the support member is spikeless and/or substantially flat. Further, the support member is formed with an electrical path for conducting current between the resilient gas diffusion layer and position(s) on the flow field plate face.

  5. Studies on the interaction between the Ehrlich ascites tumor cell and its fluid environment

    SciTech Connect

    Magnani, B.

    1984-01-01

    In this dissertation, the glycolytic nature of the Ehrlich ascites tumor (EAT) cell is disclosed both in vivo and in vitro by experiments challenging it with glucose. It is demonstrated that EAT cells can cause the extracellular pH to drop to values sufficiently acidic so as to inhibit EAT glycolysis. However, the extracellular fluid or the Ascites Supernatant Fluid (ASF) reduced the extent to which the pH dropped during EAT cell glycolysis. A comparison of the activities of the sera from tumor-bearing mice and normal mice revealed that the serumfrom the tumor-bearing mice reduced the pH fall generated by the EAT cell in the same way as did ASF; normal mouse serum had no such effect. The metabolic pathways utilized during glucose catabolism were examined by radio-respirometry and the results demonstrated that the high percentage of the glucose conversion to lactate occurred because of partial blockade of the TCA cycle. The databolism of glutamine, glutamic acid, asparagine, aspartic acid, and alanine was enhanced by ASF as determined by measuring /sup 14/CO/sub 2/ from /sup 14/C-labelled amino acids, with glutamine catabolism enhanced about three-fold. Fractionation experiments revealed that ASF contained a factor(s) responsible for this enhancement that had a molecular weight greater than 300,000 daltons and was heat-labile.

  6. Surface force confinement cell for neutron reflectometry studies of complex fluids under nano-confinement

    SciTech Connect

    Cho, Jae-Hie [ORNL; Smith, Gregory Scott [ORNL; Hamilton, William A. [ANSTO; Mulder, D. [University of California, Davis; Kuhl, T. L. [University of California, Davis; Mays, Jimmy [ORNL

    2008-01-01

    In this paper, we describe the construction of a new neutron surface force confinement cell (NSFCC). The NSFCC is equipped with hydraulically powered in-situ, temporally stable, force control system for simultaneous neutron reflectometry studies of nano-confined complex fluid systems. Test measurements with deuterated toluene confined between two opposing diblock copolymer (polystyrene + poly 2-vinylpyridine) coated quartz substrates demonstrate the capabilities of the NSFCC. With increasing hydraulically-applied force, a series of well-defined decreasing separations were observed from neutron reflectivity measurements. No noticeable changes in the hydraulic pressure used for controlling the surface separation were observed during the measurements, demonstrating the high stability of the apparatus. This newly designed NSFCC introduces a higher level of control for studies of confinement and consequent finite size effects on nano-scale structure in a variety of complex fluid and soft condensed matter systems.

  7. Shape transitions of fluid vesicles and red blood cells in capillary flows

    PubMed Central

    Noguchi, Hiroshi; Gompper, Gerhard

    2005-01-01

    The dynamics of fluid vesicles and red blood cells (RBCs) in cylindrical capillary flow is studied by using a three-dimensional mesoscopic simulation approach. As flow velocity increases, a model RBC is found to transit from a nonaxisymmetric discocyteto an axisymmetric parachute shape (coaxial with the flow axis), while a fluid vesicle is found to transit from a discocyte to a prolate ellipsoid. Both shape transitions reduce the flow resistance. The critical velocities of the shape transitions are linearly dependent on the bending rigidity and on the shear modulus of the membrane. Slipper-like shapes of the RBC model are observed around the transition velocities. Our results are in good agreement with experiments on RBCs. PMID:16186506

  8. Silicone fluid as a high-pressure medium in diamond anvil cells

    SciTech Connect

    Ragan, D.D.; Clarke, D.R. [Materials Department, College of Engineering, University of California, Santa Barbara, California 93106-5050 (United States)] [Materials Department, College of Engineering, University of California, Santa Barbara, California 93106-5050 (United States); Schiferl, D. [Los Alamos National Laboratory, Los Alamos, New Mexico 87545 (United States)] [Los Alamos National Laboratory, Los Alamos, New Mexico 87545 (United States)

    1996-02-01

    The usefulness of a silicone oil, Dow Corning 200, as a pressure medium in diamond anvil cells has been investigated. Common indicators of deviatoric stresses on ruby, such as changes in the {ital R}-line widths and the {ital R}2-{ital R}1 peak separation, show that this fluid does not deviate from hydrostaticity up to {approximately}15 GPa (150 kbar). The behavior of silicone is found to be very similar to the commonly used 4:1 methanol:ethanol mixture, while being much easier to use because of its higher viscosity. This ease of use and excellent performance makes silicone fluid a superior pressure medium. {copyright} {ital 1996 American Institute of Physics.}

  9. Kinetic Roughening in Two-Phase Fluid Flow through a Random Hele-Shaw Cell

    NASA Astrophysics Data System (ADS)

    Pauné, Eduard; Casademunt, Jaume

    2003-04-01

    A nonlocal interface equation is derived for two-phase fluid flow, with arbitrary wettability and viscosity contrast, c=(?1-?2)/(?1+?2), in a model porous medium defined as a Hele-Shaw cell with random gap b0+?b. Fluctuations of both capillary and viscous pressure are explicitly related to the microscopic quenched disorder, yielding conserved, nonconserved, and power-law correlated noise terms. Two length scales are identified that control the possible scaling regimes and which scale with capillary number Ca as ?1˜b0(cCa)-1/2 and ?2˜b0Ca-1. Exponents for forced fluid invasion are obtained from numerical simulation and compared with recent experiments.

  10. Proteolytic Activity Attenuates the Response of Endothelial Cells to Fluid Shear Stress

    PubMed Central

    Altshuler, Angelina E.; Morgan, Mary J.; Chien, Shu; Schmid-Schönbein, Geert W.

    2012-01-01

    Recent evidence indicates that several experimental pathophysiological conditions are associated with elevated protease activity in plasma, which impacts endothelial function. We hypothesize that extracellular structures bound to the endothelial cell (EC) membrane may be degraded by proteolytic activity and cause the cells to respond abnormally to physiological shear stress (12 dyn/cm2). To test this hypothesis, cultured bovine aortic endothelial cells (BAECs) were exposed to low levels of a serine protease, trypsin. Extracellular mechanosensor densities of the glycocalyx and vascular endothelial growth factor receptor 2 (VEGFR-2) were determined. Metabolic dysfunction was tested by examining insulin receptor and glucose uptake levels. Protease treatment impaired the cells’ ability to align in the direction of fluid flow after 12 hours of shear stress; however, cells realigned after an additional 12 hours of shear stress with protease inhibition. Proteases caused reduction in the densities of glycocalyx, VEGFR-2, and insulin receptor in static and shear conditions, except for static VEGFR-2 cells. Under static conditions, protease-treated endothelial cells had reduced glucose uptake compared to untreated controls. Under shear, however, glucose uptake for protease-treated BAECs was greater than untreated controls. In conclusion, protease activity in plasma alters the exofacial membrane components of ECs and may interfere with mechanotransduction. PMID:22545072

  11. Shifts in the concentrations of magnesium and calcium in early porcine and rat wound fluids activate the cell migratory response.

    PubMed Central

    Grzesiak, J J; Pierschbacher, M D

    1995-01-01

    Accruing evidence indicates that the levels of extracellular Mg2+ and Ca2+ can have a distinct impact on the adhesive and migratory activities of many cell types. The physiological relevance of these observations, however, has remained largely unexplored. In the present study, wound fluids collected throughout the early stages of cutaneous wound repair were examined for possible Mg2+ and Ca2+ fluctuations. Early in the process, when cell migration into the wound site is initiated, Mg2+ is elevated and Ca2+ is reduced (Mg2+:Ca2+ = 1). As wound healing progresses, wound fluid concentrations of Mg2+ and Ca2+ begin to return to normal plasma levels (Mg2+:Ca2+ = 0.4). When macrophages, keratinocytes, fibroblasts, and endothelial cells were exposed to dialyzed wound fluid, the migration stimulated by undialyzed wound fluid was lost. Addition back to dialyzed wound fluid of 24 h, postinjury concentrations of Mg2+ and Ca2+ restored all migratory stimulus. This observed migration is approximately twofold greater than when normal plasma Mg2+ and Ca2+ concentrations are present. Changes in the levels of Mg2+ and Ca2+ in wound fluid occur during the same period that inflammatory cells, keratinocytes, fibroblasts, and neovasculature have been shown to migrate during wound healing in vivo. Together, these data suggest that the impact of these changes on integrins and E-cadherin may play a direct role in the activation and maintenance of the migratory phenotypes of the cells involved in the wound healing process. PMID:7814620

  12. Imaging of Lactobacillus brevis single cells and microcolonies without a microscope by an ultrasensitive chemiluminescent enzyme immunoassay with a photon-counting television camera.

    PubMed

    Yasui, T; Yoda, K

    1997-11-01

    An ultrasensitive chemiluminescent enzyme immunoassay (CLEIA) was developed for the rapid detection and quantification of Lactobacillus brevis contaminants in beer and pitching yeast (Saccharomyces cerevisiae slurry collected for reinoculation). L. brevis cells trapped on a 47-mm nucleopore membrane (0.4-micron pore size) were reacted with a peroxidase-labelled Lactobacillus group E antibody and then subjected to an enhanced CLEIA analysis with 4-iodophenol as the enhancer. The combination of a nucleopore membrane with low background characteristics that enables the antigen-antibody reaction to proceed through the pores of the membrane and a labelled antibody prepared by the maleimide hinge method with minimal nonspecific binding characteristics was essential to minimize background in the detection of single cells. An ultrahigh sensitive charge-coupled device (CCD) camera equipped with a fiber optics image intensifier permitted the imaging of single cells. A clear correlation existed between the number of luminescent spots observed and the plate count [y (CLEIA) = 0.990x (plate count) + 15.9, where n = 7, r = 0.993, and P < 0.001]. Microscopic observation confirmed that the luminescent spots were produced by single cells. This assay could be used to detect approximately 20 L. brevis cells in 633 ml of beer within 4 h. Our ultrasensitive CLEIA could also be used to detect microcolonies approximately 20 microns in diameter which had formed on a membrane after 15 to 18 h of incubation. This method, which we called the microcolony immunoluminescence (MIL) method, increased the signal-to-noise ratio dramatically. The MIL method could be used to detect a 10(0) level of L. brevis contamination in 633 ml of beer and a 1/10(8) level of L. brevis contamination in pitching yeast within 1 day (15 to 18 h to form microcolonies and 2 h for CLEIA). PMID:9361439

  13. Molecular and phenotypic characterization of human amniotic fluid-derived cells: a morphological and proteomic approach.

    PubMed

    Pipino, Caterina; Pierdomenico, Laura; Di Tomo, Pamela; Di Giuseppe, Fabrizio; Cianci, Eleonora; D'Alimonte, Iolanda; Morabito, Caterina; Centurione, Lucia; Antonucci, Ivana; Mariggiò, Maria A; Di Pietro, Roberta; Ciccarelli, Renata; Marchisio, Marco; Romano, Mario; Angelucci, Stefania; Pandolfi, Assunta

    2015-06-15

    Mesenchymal Stem Cells derived from Amniotic Fluid (AFMSCs) are multipotent cells of great interest for regenerative medicine. Two predominant cell types, that is, Epithelial-like (E-like) and Fibroblast-like (F-like), have been previously detected in the amniotic fluid (AF). In this study, we examined the AF from 12 donors and observed the prevalence of the E-like phenotype in 5, whereas the F-like morphology was predominant in 7 samples. These phenotypes showed slight differences in membrane markers, with higher CD90 and lower Sox2 and SSEA-4 expression in F-like than in E-like cells; whereas CD326 was expressed only in the E-like phenotype. They did not show any significant differences in osteogenic, adipogenic or chondrogenic differentiation. Proteomic analysis revealed that samples with a predominant E-like phenotype (HC1) showed a different profile than those with a predominant F-like phenotype (HC2). Twenty-five and eighteen protein spots were differentially expressed in HC1 and HC2 classes, respectively. Of these, 17 from HC1 and 4 from HC2 were identified by mass spectrometry. Protein-interaction networks for both phenotypes showed strong interactions between specific AFMSC proteins and molecular chaperones, such as preproteasomes and mature proteasomes, both of which are important for cell cycle regulation and apoptosis. Collectively, our results provide evidence that, regardless of differences in protein profiling, the prevalence of E-like or F-like cells in AF does not affect the differentiation capacity of AFMSC preparations. This may be valuable information with a view to the therapeutic use of AFMSCs. PMID:25608581

  14. Counting whole numbers

    NSDL National Science Digital Library

    Ms. Hirst

    2007-10-12

    Identify and use whole numbers up to 100 Here are some links to help you learn more about counting Teach R Kids Math counting and number activity themes Here are some games to help you practice your counting counting cherrios Bunny Count Connect the Dots Game ...

  15. Short communication: Associations between teat dimensions and milking-induced changes in teat dimensions and quarter milk somatic cell counts in dairy cows.

    PubMed

    Zwertvaegher, I; De Vliegher, S; Verbist, B; Van Nuffel, A; Baert, J; Van Weyenberg, S

    2013-02-01

    Although many studies have examined the relation between a wide range of factors and quarter milk somatic cell count (qSCC), including physical characteristics of the teat and changes in teat tissue due to milking, the effect of short-term, milking-induced changes in teat dimensions on somatic cell count has not yet been investigated. To identify teat dimensions and milking-induced changes in teat dimensions associated with qSCC, we conducted a longitudinal study (n(herds)=6, n(cows)=72, n(measurements)=12). Parity, stage of lactation, teat barrel diameter, and changes in teat barrel diameter during milking were identified as factors associated with qSCC. Teats with wider barrels had higher qSCC. Negative changes in the diameter of the teat barrel during milking (i.e., thinner teats postmilking compared with premilking) were associated with lower qSCC, whereas positive changes (i.e., thicker teats postmilking compared with premilking) were associated with higher qSCC. Selection toward more optimal teat characteristics may therefore result in improved milk quality and udder health. However, a threshold might exist for the maximum reduction in teat barrel diameter below which udder health is negatively influenced. If so, changes in teat barrel diameter might serve as an indicator for suboptimal milking and incorrect choice of teatcup liner or milking machine settings and thus help improve management of the herd. PMID:23219124

  16. Parametric study of the vibration-induced repulsion or attraction force on a particle in a viscous fluid cell.

    PubMed

    Saadatmand, Mehrrad; Kawaji, Masahiro

    2014-04-01

    Experiments and three-dimensional direct numerical simulations were performed to investigate the effects of physical parameters on the repulsion or attraction force affecting the motion of a particle oscillating near a solid wall of a fluid cell under microgravity. The following physical parameters were investigated: fluid cell amplitude, fluid and particle densities, angular frequency of the cell vibration, initial distance between the particle centroid and the closest cell wall, particle radius, and dynamic viscosity. Based on the simulations, a nondimensional relation was developed to relate those physical parameters to the repulsion or attraction force affecting the particle. The relation shows that the repulsion or attraction force is increased by the increase in the cell vibration amplitude and frequency and also the force direction would change from attraction to repulsion above a threshold fluid viscosity. Relations to other physical parameters were also studied and are reported. This paper follows our previous work on the physical mechanism of observed repulsion force on a particle in a viscous fluid cell [M. Saadatmand and M. Kawaji, Phys. Rev. E 88, 023019 (2013)]. PMID:24827334

  17. Air sparging for prevention of antibody disulfide bond reduction in harvested CHO cell culture fluid.

    PubMed

    Mun, Melissa; Khoo, Stefanie; Do Minh, Aline; Dvornicky, James; Trexler-Schmidt, Melody; Kao, Yung-Hsiang; Laird, Michael W

    2015-04-01

    During the scale-up of several Chinese Hamster Ovary (CHO) cell monoclonal antibody production processes, significant reduction of the antibody interchain disulfide bonds was observed. The reduction was correlated with excessive mechanical cell shear during the harvest operations. These antibody reduction events resulted in failed product specifications and the subsequent loss of the drug substance batches. Several methods were recently developed to prevent antibody reduction, including modifying the cell culture media, using pre- and post-harvest chemical additions to the cell culture fluid (CCF), lowering the pH, and air sparging of the harvested CCF (HCCF). The work described in this paper further explores the option of HCCF air sparging for preventing antibody reduction. Here, a small-scale model was developed using a 3-L bioreactor to mimic the conditions of a manufacturing-scale harvest vessel and was subsequently employed to evaluate several air sparging strategies. In addition, these studies enabled further understanding of the relationships between cell lysis levels, oxygen consumption, and antibody reduction. Finally, the effectiveness of air sparging for several CHO cell lines and the potential impact on product quality were assessed to demonstrate that air sparging is an effective method in preventing antibody reduction. PMID:25384896

  18. Avian leucocyte counting using the hemocytometer

    USGS Publications Warehouse

    Dein, F.J.; Wilson, A.; Fischer, D.; Langenberg, P.

    1994-01-01

    Automated methods for counting leucocytes in avian blood are not available because of the presence of nucleated erythrocytes and thrombocytes. Therefore, total white blood cell counts are performed by hand using a hemocytometer. The Natt and Herrick and the Unopette methods are the most common stain and diluent preparations for this procedure. Replicate hemocytometer counts using these two methods were performed on blood from four birds of different species. Cells present in each square of the hemocytometer were counted. Counting cells in the corner, side, or center hemocytometer squares produced statistically equivalent results; counting four squares per chamber provided a result similar to that obtained by counting nine squares; and the Unopette method was more precise for hemocytometer counting than was the Natt and Herrick method. The Unopette method is easier to learn and perform but is an indirect process, utilizing the differential count from a stained smear. The Natt and Herrick method is a direct total count, but cell identification is more difficult.

  19. Kinetics of T-cell-based assays on cerebrospinal fluid and peripheral blood mononuclear cells in patients with tuberculous meningitis

    PubMed Central

    Park, Ki-Ho; Lee, Mi Suk; Lee, Sang-Oh; Choi, Sang-Ho; Kim, Yang Soo; Woo, Jun Hee; Kang, Joong Koo; Lee, Sang-Ahm

    2014-01-01

    Background/Aims The goal of this study was to monitor tuberculosis (TB)-specific T-cell responses in cerebrospinal fluid-mononuclear cells (CSF-MCs) and peripheral blood mononuclear cells (PBMCs) in patients with tuberculous meningitis (TBM) over the course of anti-TB therapy. Methods Adult patients (? 16 years) with TBM admitted to Asan Medical Center, Seoul, South Korea, were prospectively enrolled between April 2008 and April 2011. Serial blood or CSF samples were collected over the course of the anti-TB therapy, and analyzed using an enzyme-linked immunosorbent spot (ELISPOT) assay. Results Serial ELISPOT assays were performed on PBMCs from 17 patients (seven definite, four probable, and six possible TBM) and CSF-MC from nine patients (all definite TBM). The median number of interferon-gamma (IFN-?)-producing T-cells steadily increased during the first 6 months after commencement of anti-TB therapy in PBMCs. Serial CSF-MC ELISPOT assays revealed significant variability in immune responses during the first 6 weeks of anti-TB therapy, though early increases in CSF-MC ELISPOT results were associated with treatment failure or paradoxical response. Conclusions Serial analysis of PBMCs by ELISPOT during the course of treatment was ineffective for predicting clinical response. However, increases in TB-specific IFN-?-producing T-cells in CSF-MC during the early phase of anti-TB therapy may be predictive of clinical failure. PMID:25378978

  20. Monocyte count at diagnosis is a prognostic parameter in diffuse large B-cell lymphoma: results from a large multicenter study involving 1191 patients in the pre- and post-rituximab era

    PubMed Central

    Tadmor, Tamar; Bari, Alessia; Sacchi, Stefano; Marcheselli, Luigi; Liardo, Eliana Valentina; Avivi, Irit; Benyamini, Noam; Attias, Dina; Pozzi, Samantha; Cox, Maria Christina; Baldini, Luca; Brugiatelli, Maura; Federico, Massimo; Polliack, Aaron

    2014-01-01

    In this study we assessed the prognostic significance of absolute monocyte count and selected the best cut-off value at diagnosis in a large cohort of patients with diffuse large B-cell lymphoma. Data were retrieved for therapy-naïve patients with diffuse large B-cell lymphoma followed in Israel and Italy during 1993–2010. A final cohort of 1017 patients was analyzed with a median follow up of 48 months and a 5-year overall survival rate of 68%. The best absolute monocyte count cut-off level was 630/mm3 and the 5-year overall survival for patients with counts below this cut-off was 71%, whereas it was 59% for those with a count >630 mm3 (P=0.0002). Of the 1017 patients, 521 (51%) were treated with chemo-immunotherapy, and in this cohort, using multivariate analysis, elevated monocyte count retained a negative prognostic value even when adjusted for International Prognostic Index (HR1.54, P=0.009). This large study shows that a simple parameter such as absolute monocyte count (>630/mm3) can easily be used routinely in the evaluation of newly diagnosed diffuse large B-cell lymphoma to identify high-risk patients with a worse survival in the rituximab era. PMID:23935023

  1. Dendritic cells in synovial fluid of chronic inflammatory arthritis lack CD80 surface expression.

    PubMed Central

    Summers, K L; Daniel, P B; O'Donnell, J L; Hart, D N

    1995-01-01

    Dendritic cells (DC) act as potent primary antigen-presenting cells in many immune responses and therefore may have a role in the initiation and perpetuation of the synovial inflammation in chronic inflammatory arthritis. To examine their function, it is important to isolate fresh DC from arthritic joints without aberrant activation. We have developed a technique using minimal cell manipulation to isolate DC from the synovial fluid of chronic arthritic patients. Using this method, DC were shown to be potent allostimulatory cells, with 63-90% of cells lacking lineage-specific markers (lin-), but positive for MHC class II molecules. Two morphologically distinct populations of these cells were identified in 10 out of 13 DC preparations. Both populations expressed CD40, intercellular adhesion molecule-1 (ICAM-1), ICAM-2, ICAM-3 and leucocyte function associated antigen-3 (LFA-3), but the predominant population, which was larger and more typical of cultured blood DC, had a higher density of these antigens compared with the minor population, which were smaller and morphologically similar to lymphocytes. Two new MoAbs which label activated human blood DC, HB15 (CD83) and CMRF-44, were tested. CD83 labelled very weakly or not at all, whereas CMRF-44 was positive on the larger cells only. Likewise, the costimulator molecule, B7/BB1 (CD80), was not detected on the surface of either synovial lin- cell population, reverse transcriptase polymerase chain reaction (RT-PCR) showed little or no CD80 mRNA, and no binding of the CTLA-4Ig fusion protein was found. These results suggest that synovial DC are not, despite the inflammatory environment, in a fully activated state. Images Fig. 2 Fig. 6 PMID:7535211

  2. Coupled thermal, electrical, and fluid flow analyses of AMTEC multitube cell with adiabatic side wall

    SciTech Connect

    Schock, A.; Or, C. [Orbital Sciences Corporation 20301 Century Blvd. Germantown, Maryland20874 (United States); Noravian, H. [ANALYTIX Corporation Timonium, Maryland (United States)

    1997-01-01

    The paper describes a novel OSC-generated methodology for analyzing the performance of multitube AMTEC (Alkali Metal Thermal-to-Electrical Conversion) cells, which are under development by AMPS (Advanced Modular Power Systems, Inc.) for the Air Force Phillips Laboratory (AFPL) and NASA{close_quote}s Jet Propulsion Laboratory (JPL), for possible application to the Pluto Express and other space missions. The OSC study was supported by the Department of Energy (DOE), and was strongly encouraged by JPL, AFPL, and AMPS. It resulted in an iterative procedure for the coupled solution of the interdependent thermal, electrical, and fluid flow differential and integral equations governing the performance of AMTEC cells and generators. The paper clarifies the OSC procedure by presenting detailed results of its application to an illustrative example of a converter cell with an adiabatic side wall, including the non-linear axial variation of temperature, pressure, open-circuit voltage, interelectrode voltage, current density, axial current, sodium mass flow, and power density. The next paper in these proceedings describes parametric results obtained by applying the same procedure to variations of the baseline adiabatic converter design, culminating in an OSC-recommended revised cell design. A subsequent paper in these proceedings extends the procedure to analyze a variety of OSC-designed radioisotope-heated generators employing non-adiabatic multitube AMTEC cells. {copyright} {ital 1997 American Institute of Physics.}

  3. Coupled thermal, electrical, and fluid flow analyses of AMTEC multitube cell with adiabatic side wall

    NASA Astrophysics Data System (ADS)

    Schock, A.; Or, C.; Noravian, H.

    1997-01-01

    The paper describes a novel OSC-generated methodology for analyzing the performance of multitube AMTEC (Alkali Metal Thermal-to-Electrical Conversion) cells, which are under development by AMPS (Advanced Modular Power Systems, Inc.) for the Air Force Phillips Laboratory (AFPL) and NASA's Jet Propulsion Laboratory (JPL), for possible application to the Pluto Express and other space missions. The OSC study was supported by the Department of Energy (DOE), and was strongly encouraged by JPL, AFPL, and AMPS. It resulted in an iterative procedure for the coupled solution of the interdependent thermal, electrical, and fluid flow differential and integral equations governing the performance of AMTEC cells and generators. The paper clarifies the OSC procedure by presenting detailed results of its application to an illustrative example of a converter cell with an adiabatic side wall, including the non-linear axial variation of temperature, pressure, open-circuit voltage, interelectrode voltage, current density, axial current, sodium mass flow, and power density. The next paper in these proceedings describes parametric results obtained by applying the same procedure to variations of the baseline adiabatic converter design, culminating in an OSC-recommended revised cell design. A subsequent paper in these proceedings extends the procedure to analyze a variety of OSC-designed radioisotope-heated generators employing non-adiabatic multitube AMTEC cells.

  4. Three-Dimensional Computational Fluid Dynamics Modeling of Solid Oxide Electrolysis Cells and Stacks

    SciTech Connect

    Grant Hawkes; James O'Brien; Carl Stoots; Stephen Herring

    2008-07-01

    A three-dimensional computational fluid dynamics (CFD) electrochemical model has been created for detailed analysis of a high-temperature electrolysis stack (solid oxide fuel cells operated as electrolyzers). Inlet and outlet plenum flow distributions are discussed. Maldistribution of plena flow show deviations in per-cell operating conditions due to non-uniformity of species concentrations. Models have also been created to simulate experimental conditions and for code validation. Comparisons between model predictions and experimental results are discussed. Mass, momentum, energy, and species conservation and transport are provided via the core features of the commercial CFD code FLUENT. A solid-oxide fuel cell (SOFC) model adds the electrochemical reactions and loss mechanisms and computation of the electric field throughout the cell. The FLUENT SOFC user-defined subroutine was modified for this work to allow for operation in the electrolysis mode. Model results provide detailed profiles of temperature, Nernst potential, operating potential, activation over-potential, anode-side gas composition, cathode-side gas composition, current density and hydrogen production over a range of stack operating conditions. Variations in flow distribution, and species concentration are discussed. End effects of flow and per-cell voltage are also considered. Predicted mean outlet hydrogen and steam concentrations vary linearly with current density, as expected. Contour plots of local electrolyte temperature, current density, and Nernst potential indicate the effects of heat transfer, reaction cooling/heating, and change in local gas composition.

  5. Electrical Impedance Study of Cultured Endothelial Cells Under Fluid Shear Stress

    NASA Astrophysics Data System (ADS)

    Dong, Chunzhi; Depaola, Natacha; Keese, Charles R.; Giaever, Ivar

    2004-03-01

    The lumen of blood vessels is lined with a monolayer of endothelial cells (EC). In this work, electric cell-substrate impedance sensing (ECIS) was used to monitor the effect of fluid shear stress (FSS) on the morphology and function of cultured EC layers. Confluent layers of bovine aortic endothelial cells (BAEC) were grown on small gold electrodes and exposed to different flow conditions, while the impedance of the system was monitored. When the cells are subjected to FSS, the impedance rapidly increases 5-10%, and if the FSS is removed after a few minutes duration, the impedance returns back to the initial level in about 10 minutes. If the FSS remains for a long duration, the impedance will decrease 20-30% over a 10-15 hour period but ultimately returns to the original value, as the cells apparently accommodate to the FSS condition. These results suggest that ECIS may provide a sensitive means to study the response of EC to shear stress in vitro.

  6. Stem cells from foetal adnexa and fluid in domestic animals: an update on their features and clinical application.

    PubMed

    Iacono, E; Rossi, B; Merlo, B

    2015-06-01

    Over the past decade, stem cell research has emerged as an area of major interest for its potential in regenerative medicine applications. This is in constant need of new cell sources to conceive regenerative medicine approaches for diseases that are still without therapy. Scientists drew the attention towards alternative sources such as foetal adnexa and fluid, as these sources possess many advantages: first of all, cells can be extracted from discarded foetal material and it is non-invasive and inexpensive for the patient; secondly, abundant stem cells can be obtained; and finally, these stem cell sources are free from ethical considerations. Cells derived from foetal adnexa and fluid preserve some of the characteristics of the primitive embryonic layers from which they originate. Many studies have demonstrated the differentiation potential in vitro and in vivo towards mesenchymal and non-mesenchymal cell types; in addition, the immune-modulatory properties make these cells a good candidate for allo- and xenotransplantation. Naturally occurring diseases in domestic animals can be more ideal as disease model of human genetic and acquired diseases and could help to define the potential therapeutic use efficiency and safety of stem cells therapies. This review offers an update on the state of the art of characterization of domestic animals' MSCs derived from foetal adnexa and fluid and on the latest findings in pre-clinical or clinical setting of the stem cell populations isolated from these sources. PMID:25703812

  7. TTF-1 and napsin A on cell blocks and supernatants of pleural fluids for labeling malignant effusions.

    PubMed

    Porcel, José M; Palma, Rosa; Bielsa, Silvia; Esquerda, Aureli; Gatius, Sonia; Matias-Guiu, Xavier; Salud, Antonieta

    2015-07-01

    In this retrospective study of 80 pleural effusions, the combination of thyroid transcription factor 1 (TTF-1) and napsin A immunostaining on fluid cell blocks was positive in 80% of lung adenocarcinomas. Although measuring TTF-1 pleural fluid concentrations was of no value, quantification of napsin A levels allowed the identification of one third of the double-negative stained lung adenocarcinomas, with an overall accuracy similar to classical tumour markers for malignant-benign discrimination (sensitivity 40%, specificity 100%). PMID:25873201

  8. Preparation of Japanese encephalitis virus nonstructural protein NS1 obtained from culture fluid of JEV-infected Vero cells

    Microsoft Academic Search

    T. Lee; K. Watanabe; C. Aizawa; A. Nomoto; H. Hashimoto

    1991-01-01

    Summary The Japanese encephalitis virus (JEV) nonstructural protein NS1 was released efficiently into culture fluid of JEV-infected Vero cells. The JEV NS1 protein in the infected culture fluid was found almost as a high-molecular-weight form, probably a dimer form of NS1, and was converted to a monomer by boiling. Large amounts of NS1 protein were accumulated in the infected culture

  9. Baseline CD4+ T Cell Counts Correlates with HIV-1 Synonymous Rate in HLA-B*5701 Subjects with Different Risk of Disease Progression

    PubMed Central

    Norström, Melissa M.; Veras, Nazle M.; Huang, Wei; Proper, Mattia C. F.; Cook, Jennifer; Hartogensis, Wendy; Hecht, Frederick M.

    2014-01-01

    HLA-B*5701 is the host factor most strongly associated with slow HIV-1 disease progression, although risk of progression may vary among patients carrying this allele. The interplay between HIV-1 evolutionary rate variation and risk of progression to AIDS in HLA-B*5701 subjects was studied using longitudinal viral sequences from high-risk progressors (HRPs) and low-risk progressors (LRPs). Posterior distributions of HIV-1 genealogies assuming a Bayesian relaxed molecular clock were used to estimate the absolute rates of nonsynonymous and synonymous substitutions for different set of branches. Rates of viral evolution, as well as in vitro viral replication capacity assessed using a novel phenotypic assay, were correlated with various clinical parameters. HIV-1 synonymous substitution rates were significantly lower in LRPs than HRPs, especially for sets of internal branches. The viral population infecting LRPs was also characterized by a slower increase in synonymous divergence over time. This pattern did not correlate to differences in viral fitness, as measured by in vitro replication capacity, nor could be explained by differences among subjects in T cell activation or selection pressure. Interestingly, a significant inverse correlation was found between baseline CD4+ T cell counts and mean HIV-1 synonymous rate (which is proportional to the viral replication rate) along branches representing viral lineages successfully propagating through time up to the last sampled time point. The observed lower replication rate in HLA-B*5701 subjects with higher baseline CD4+ T cell counts provides a potential model to explain differences in risk of disease progression among individuals carrying this allele. PMID:25187947

  10. Fluid biopsy for circulating tumor cell identification in patients with early-and late-stage non-small cell lung cancer: a glimpse into lung cancer biology

    Microsoft Academic Search

    Marco Wendel; Lyudmila Bazhenova; Rogier Boshuizen; Anand Kolatkar; Meghana Honnatti; Edward H Cho; Dena Marrinucci; Ajay Sandhu; Anthony Perricone; Patricia Thistlethwaite; Kelly Bethel; Jorge Nieva; Michel van den Heuvel; Peter Kuhn

    2012-01-01

    Circulating tumor cell (CTC) counts are an established prognostic marker in metastatic prostate, breast and colorectal cancer, and recent data suggest a similar role in late stage non-small cell lung cancer (NSCLC). However, due to sensitivity constraints in current enrichment-based CTC detection technologies, there are few published data about CTC prevalence rates and morphologic heterogeneity in early-stage NSCLC, or the

  11. Usefulness of Cellular Analysis of Bronchoalveolar Lavage Fluid for Predicting the Etiology of Pneumonia in Critically Ill Patients

    PubMed Central

    Hong, Hyo-Lim; Kim, Sung-Han; Huh, Jin Won; Sung, Heungsup; Lee, Sang-Oh; Kim, Mi-Na; Jeong, Jin-Yong; Lim, Chae-Man; Kim, Yang Soo; Woo, Jun Hee; Koh, Younsuck

    2014-01-01

    Background The usefulness of bronchoalveolar lavage (BAL) fluid cellular analysis in pneumonia has not been adequately evaluated. This study investigated the ability of cellular analysis of BAL fluid to differentially diagnose bacterial pneumonia from viral pneumonia in adult patients who are admitted to intensive care unit. Methods BAL fluid cellular analysis was evaluated in 47 adult patients who underwent bronchoscopic BAL following less than 24 hours of antimicrobial agent exposure. The abilities of BAL fluid total white blood cell (WBC) counts and differential cell counts to differentiate between bacterial and viral pneumonia were evaluated using receiver operating characteristic (ROC) curve analysis. Results Bacterial pneumonia (n?=?24) and viral pneumonia (n?=?23) were frequently associated with neutrophilic pleocytosis in BAL fluid. BAL fluid median total WBC count (2,815/µL vs. 300/µL, P<0.001) and percentage of neutrophils (80.5% vs. 54.0%, P?=?0.02) were significantly higher in the bacterial pneumonia group than in the viral pneumonia group. In ROC curve analysis, BAL fluid total WBC count showed the best discrimination, with an area under the curve of 0.855 (95% CI, 0.750–0.960). BAL fluid total WBC count ?510/µL had a sensitivity of 83.3%, specificity of 78.3%, positive likelihood ratio (PLR) of 3.83, and negative likelihood ratio (NLR) of 0.21. When analyzed in combination with serum procalcitonin or C-reactive protein, sensitivity was 95.8%, specificity was 95.7%, PLR was 8.63, and NLR was 0.07. BAL fluid total WBC count ?510/µL was an independent predictor of bacterial pneumonia with an adjusted odds ratio of 13.5 in multiple logistic regression analysis. Conclusions Cellular analysis of BAL fluid can aid early differential diagnosis of bacterial pneumonia from viral pneumonia in critically ill patients. PMID:24824328

  12. Frequency-dependent properties of a fluid jet stimulus: calibration, modeling, and application to cochlear hair cell bundles.

    PubMed

    Dinklo, Theo; Meulenberg, Cécil J W; van Netten, Sietse M

    2007-06-01

    The investigation of small physiological mechano-sensory systems, such as hair cells or their accessory structures in the inner ear or lateral line organ, requires mechanical stimulus equipment that allows spatial manipulation with micrometer precision and stimulation with amplitudes down to the nanometer scale. Here, we describe the calibration of a microfluid jet produced by a device that was designed to excite individual cochlear hair cell bundles or cupulae of the fish superficial lateral line system. The calibration involves a precise definition of the linearity and time- and frequency-dependent characteristics of the fluid jet as produced by a pressurized fluid-filled container combined with a glass pipette having a microscopically sized tip acting as an orifice. A procedure is described that can be applied during experiments to obtain a fluid jet's frequency response, which may vary with each individual glass pipette. At small orifice diameters (<15 mum), the fluid velocity of the jet is proportional to the displacement of the piezoelectric actuator pressurizing the container's volume and is suitable to stimulate the hair bundles of sensory hair cells. With increasing diameter, the fluid jet velocity becomes proportional to the actuator's velocity. The experimentally observed characteristics can be described adequately by a dynamical model of damped fluid masses coupled by elastic components. PMID:17387553

  13. Characterization and hepatogenic differentiation of mesenchymal stem cells from human amniotic fluid and human bone marrow: A comparative study

    Microsoft Academic Search

    Yu-Bao Zheng; Zhi-Liang Gao; Chan Xie; Hai-Peng Zhu; Liang Peng; Jun-Hong Chen; Yu Tian Chong

    2008-01-01

    Since stem cells can differentiate into hepatocyte, stem cell-based therapy becomes a potential alternative treatment for terminal liver diseases. However, an appropriate source of human mesenchymal stem cells (hMSCs) for hepatocytes has not yet been clearly elucidated. The aim of the present study was to investigate the in vitro biological characterization and hepatic differentiation potential of human amniotic fluid-derived mesenchymal

  14. Somatic cell count and milk neutrophil viability of dairy heifers with specific CXCR1 genotypes following experimental intramammary infection with Staphylococcus chromogenes originating from milk.

    PubMed

    Verbeke, Joren; Piccart, Kristine; Piepers, Sofie; Van Poucke, Mario; Peelman, Luc; De Visscher, Anneleen; De Vliegher, Sarne

    2015-06-01

    Previous observational studies suggest an association between polymorphism c.980A>G in the CXCR1 gene, encoding the chemokine (C-X-C motif) receptor 1, and the innate immunity and infection status of the mammary gland. Mammary glands of eight Holstein heifers were experimentally infected with a Staphylococcus chromogenes isolate originating from a chronic intramammary infection (IMI) to study differences between CXCR1 genotypes c.980AG and c.980GG. Quarters from heifers with genotypes c.980AG and c.980GG developed subclinical mastitis but showed differences in the early response at 6-18?h post challenge. Bacterial count at 18?h post challenge tended to be higher in quarters from c.980AG heifers compared to c.980GG heifers. Somatic cell count (SCC) was higher at 6?h post challenge and tended to be higher at 9?h post challenge in c.980AG heifers compared to c.980GG heifers. Milk production decreased similarly. Milk neutrophils of c.980AG heifers showed more apoptosis at 9?h post challenge and tended to show more necrosis at 6, 9 and 12?h post challenge than c.980GG heifers. Differences were less pronounced in the later stage (>18?h) of infection. The results demonstrate that CXCR1 polymorphism can influence SCC and milk neutrophil viability following experimental IMI. PMID:25933826

  15. Recurrence and death in non-small cell lung carcinomas: a prognostic model using pathological parameters, microvessel count, and gene protein products.

    PubMed

    Fontanini, G; Vignati, S; Bigini, D; Mussi, A; Lucchi, M; Chiné, S; Angeletti, C A; Bevilacqua, G

    1996-06-01

    The 5-year survival rate of non-small cell lung carcinoma (NSCLC) has only marginally improved during the past two decades, despite advances in surgery and chemoradiotherapy. Major efforts are currently directed toward biological characterization of these tumors to define biomarkers able to add further prognostic information, thus improving new therapeutic protocols. We analyzed the predictive relevance of the microvessel count (MC), bcl-2 and p53 proteins, proliferative activity, and usual postsurgical parameters on recurrence and overall survival in a series of 70 patients with NSCLC. The expression of biological parameters (p53, bcl-2, proliferative activity, and MC) was detected using immunohistochemistry on paraffin-embedded and frozen sections from the tumors treated with surgical resection alone until relapse. In the univariate analysis, the histotype, tumor status, node status, p53, bcl-2, and MC have been shown to significantly affect progression and death. In the multiple logistic regression analysis, the MC (P < 0.000001), tumor status (P < 0.005), and node status (P < 0.0002) influenced the overall survival while prediction of relapse was strongly revealed by tumor status (P < 0.005), nodal metastatic involvement (P < 0.000001), and the assessment of the vascular count (P < 0.0004). These data have allowed the creation of a multivariate model which may add more information on risk of recurrence and death in patients with NSCLC and can form the basis for future randomized clinical trials. PMID:9816269

  16. An efficient and simple method for measuring (226)Ra using the scintillation cell in a delayed coincidence counting system (RaDeCC).

    PubMed

    Waska, Hannelore; Kim, Seolwon; Kim, Guebuem; Peterson, Richard N; Burnett, William C

    2008-12-01

    A delayed coincidence counter (RaDeCC), developed to determine ultra-low levels of (223)Ra (half life = 11.1 days) and (224)Ra (half life = 3.6 days) in seawater, was adapted to measure (226)Ra (half life = 1622 years). After pre-concentration of Ra from seawater onto MnO(2)-coated fiber we show in this study that the (226)Ra activity can be determined using the RaDeCC's ability to record alpha decay of its daughters as total counts. For sufficient ingrowth of (222)Rn, the Mn-fiber is hermetically sealed in a column for a few days. Then, the ingrown (222)Rn is circulated through the RaDeCC air-loop system followed by shutting down of the pump and closure of the scintillation cell for equilibration. Counting may be completed within a few hours for seawater samples. Sample measurements with this method agreed well with data obtained using gamma-ray spectrometry. This proves that a set of Ra isotopes ((223)Ra, (224)Ra, and (226)Ra), commonly used for geophysical studies such as mixing rates of different water masses and submarine groundwater discharge, can be efficiently and rapidly measured using the RaDeCC. PMID:18950907

  17. The effect of cigarettes smoking on the blood counts of T and NK cells in subjects with occupational exposure to organic solvents.

    PubMed

    Moszczy?ski, P; Rutowski, J; S?owi?ski, S

    1996-09-01

    The study was carried out in a population of 139 men, divided into two control groups: 40 non-smokers and 39 smokers not exposed to chemical compounds, and two groups exposed to them: 19 non-smokers and 41 cigarette smokers with occupational contact with organic solvents. The results of toxicological analyses of air and chromatographic analyses of solvents demonstrated the presence of benzene, toluene, xylene and their partly hydrogenated derivatives, paraffin hydrocarbons, oleins, naphthenes (components of painter's naphtha), monohydric and polyhydric alcohols (butanol, cyclohexanol, butylene glycol) esters (ethyleneglycol acetate, butyl acetate) and ketones (methylisobutyl ketone, cyclohexanone). In the time of the studies the TWA values for benzene were 0 to 38 mg x m-3 (0 to 12 ppm), with arithmetic mean averages of about 19 mg x m-3 (6 ppm) and for the level of benzene 0-351 mg x m-3 (0-110 ppm) with arithmetic mean annual averages of about 48 mg x m-3 (15 ppm). Mean phenol concentration in the urine of the workers in groups I, II, III and IV respectively was: 7.9 +/- 3.5; 10.0 +/- 5.8; 16.8 +/- 6.2 and 18.4 +/- 9.7 mg x l-1. Hippuric acid concentration in the urine of the workers in groups I to IV was: 496 +/- 326, 538 +/- 341, 982 +/- 420 and 1107 +/- 507 mg x l-1 respectively. The absolute counts were determined of T-cells (CD 3+), T-helper (CD 4+), T-suppressor (CD 8+) cells and natural killer (NK) cells (CD 16+) in the peripheral blood by indirect immunofluorescence. In the subjects with occupational exposure to organic solvents the counts of T-cells and NK-cells were reduced, and the number of T-suppressor cells was raised which resulted in a decrease of the T-helper/T-suppressor ratio. These changes were more pronounced in cigarette smokers. The assessment of the immunotoxic effect of organic solvents during occupational exposure should take into consideration the possibility of a synergistic action with tobacco and may be of practical use for monitoring the toxic effect of organic solvents on the lymphocyte system. PMID:8884050

  18. Differences in cerebrospinal fluid inflammatory cell reaction of patients with leptomeningeal involvement by lymphoma and carcinoma.

    PubMed

    Illán, Julia; Simo, Marta; Serrano, Cristina; Castañón, Susana; Gonzalo, Raquel; Martínez-García, María; Pardo, Javier; Gómez, Lidia; Navarro, Miguel; Altozano, Javier Pérez; Alvarez, Ruth; Bruna, Jordi; Subirá, Dolores

    2014-12-01

    Dissemination of neoplastic cells into the cerebrospinal fluid (CSF) and leptomeninges is a devastating complication in patients with epithelial cell neoplasia (leptomeningeal carcinomatosis [LC]) and lymphomas (lymphomatous meningitis [LyM]). Information about the surrounding inflammatory cell populations is scarce. In this study, flow cytometry immunophenotyping was used to describe the distribution of the main leukocyte populations in the CSF of 83 patients diagnosed with neoplastic meningitis (LC, n = 65; LyM, n = 18). These data were compared with those obtained in the CSF from 55 patients diagnosed with the same groups of neoplasia without meningeal involvement (solid tumors, n = 36; high-grade lymphoma, n = 19). Median (interquartile) rates of lymphocytes, monocytes, and polymorphonuclear (PMN) cells were 59.7% (range, 35-76.6%), 24% (range, 16-53%), and 1.5% (range, 0-7.6%) in LC, respectively, and 98.5% (range, 70.8-100%), 1.5% (range, 0-29.3%), and 0% in LyM, respectively (P < 0.001). No difference was observed between patients with breast adenocarcinoma (n = 30) and lung adenocarcinoma (n = 21), nor with different rates of malignant CSF involvement. Patients with lymphoma (with or without LyM) had a similar CSF leukocyte distribution, but cancer patients with LC and without LC had a distinctive PMN cell rate (P = 0.002). These data show that CSF samples from patients with LC have a greater number of inflammatory cells and a different leukocyte distribution than seen in the CSF from patients with LyM. Description of PMN cells is a distinctive parameter of patients with LC, compared with the CSF from patients with LyM and patients with cancer but without LC. PMID:24746871

  19. Microbial Metabolism in Serpentinite Fluids

    NASA Astrophysics Data System (ADS)

    Crespo-Medina, M.; Brazelton, W. J.; Twing, K. I.; Kubo, M.; Hoehler, T. M.; Schrenk, M. O.

    2013-12-01

    Serpentinization is the process in which ultramafic rocks, characteristic of the upper mantle, react with water liberating mantle carbon and reducing power to potenially support chemosynthetic microbial communities. These communities may be important mediators of carbon and energy exchange between the deep Earth and the surface biosphere. Our work focuses on the Coast Range Ophiolite Microbial Observatory (CROMO) in Northern California where subsurface fluids are accessible through a series of wells. Preliminary analyses indicate that the highly basic fluids (pH 9-12) have low microbial diversity, but there is limited knowledge about the metabolic capabilities of these communties. Metagenomic data from similar serpentine environments [1] have identified Betaproteobacteria belonging to the order Burkholderiales and Gram-positive bacteria from the order Clostridiales as key components of the serpentine microbiome. In an effort to better characterize the microbial community, metabolism, and geochemistry at CROMO, fluids from two representative wells (N08B and CSWold) were sampled during recent field campaigns. Geochemical characterization of the fluids includes measurements of dissolved gases (H2, CO, CH4), dissolved inorganic and organic carbon, volatile fatty acids, and nutrients. The wells selected can be differentiated in that N08B had higher pH (10-11), lower dissolved oxygen, and cell counts ranging from 105-106 cells mL-1 of fluid, with an abundance of the betaproteobacterium Hydrogenophaga. In contrast, fluids from CSWold have slightly lower pH (9-9.5), DO, and conductivity, as well as higher TDN and TDP. CSWold fluid is also characterized for having lower cell counts (~103 cells mL-1) and an abundance of Dethiobacter, a taxon within the phylum Clostridiales. Microcosm experiments were conducted with the purpose of monitoring carbon fixation, methanotrophy and metabolism of small organic compounds, such as acetate and formate, while tracing changes in fluid chemistry and microbial community composition. These experiments are expected to provide insight into the biogeochemical dynamics of the serpentinite subsurface at CROMO and represent a first step for developing metatranscriptomic and RNA-based Stable Isotope Probing (RNA-SIP) experiments to trace microbial activity at this site. [1] Brazelton et al. (2012) Frontiers in Microbiology 2:268

  20. Stem cells from human amniotic fluid exert immunoregulatory function via secreted indoleamine 2,3-dioxygenase1.

    PubMed

    Romani, Rita; Pirisinu, Irene; Calvitti, Mario; Pallotta, Maria Teresa; Gargaro, Marco; Bistoni, Giovanni; Vacca, Carmine; Di Michele, Alessandro; Orabona, Ciriana; Rosati, Jessica; Pirro, Matteo; Giovagnoli, Stefano; Matino, Davide; Prontera, Paolo; Rosi, Gabriella; Grohmann, Ursula; Talesa, Vincenzo N; Donti, Emilio; Puccetti, Paolo; Fallarino, Francesca

    2015-07-01

    Although human amniotic fluid does contain different populations of foetal-derived stem cells, scanty information is available on the stemness and the potential immunomodulatory activity of in vitro expanded, amniotic fluid stem cells. By means of a methodology unrequiring immune selection, we isolated and characterized different stem cell types from second-trimester human amniotic fluid samples (human amniotic fluid stem cells, HASCs). Of those populations, one was characterized by a fast doubling time, and cells were thus designated as fHASCs. Cells maintained their original phenotype under prolonged in vitro passaging, and they were able to originate embryoid bodies. Moreover, fHASCs exhibited regulatory properties when treated with interferon (IFN)-?, including induction of the immunomodulatory enzyme indoleamine 2,3-dioxygenase 1 (IDO1). On coculture with human peripheral blood mononuclear cells, IFN-?-treated fHASCs caused significantly decreased T-cell proliferation and increased frequency in CD4(+)  CD25(+)  FOXP3(+) regulatory T cells. Both effects required an intact IDO1 function and were cell contact-independent. An unprecedented finding in our study was that purified vesicles from IFN-?-treated fHASCs abundantly expressed the functional IDO1 protein, and those vesicles were endowed with an fHASC-like regulatory function. In vivo, fHASCs were capable of immunoregulatory function, promoting allograft survival in a mouse model of allogeneic skin transplantation. This was concurrent with the expansion of CD4(+)  CD25(+)  Foxp3(+) T cells in graft-draining lymph nodes from recipient mice. Thus fHASCs, or vesicles thereof, may represent a novel opportunity for immunoregulatory maneuvers both in vitro and in vivo. PMID:25783564