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Sample records for fluid cell count

  1. Low accuracy of manual white blood cell count in amniotic fluid.

    PubMed

    McMillen, E; Bautista, J; Sireci, A; Kratz, A; Stotler, B

    2013-05-01

    Intra-amniotic infection (IAI) is a common cause of pre-term labour. Manual WBC count on amniotic fluid (AF) has been suggested as a diagnostic test for IAI using a threshold of 50 cells/mm(3). However, no validation studies assessing the accuracy of this method have been performed. AF samples were selected for cell count analysis. WBCs were introduced to 47 AF samples. The results from two technologists' counts were compared with the calculated expected value for WBCs in these samples. Results showed that a comparison between the technologists' WBC count to the expected WBC count yielded R(2) coefficients of 0.62 and 0.78, indicating moderate accuracy. Percentage agreement between the technologists was 67%, indicating low reproducibility. It was concluded that there was moderate correlation between the manual and the expected WBC in the spiked AF samples. Clinicians should be aware of the inaccuracy and imprecision associated with this test when evaluating a patient for IAI.

  2. White Blood Cell Count

    MedlinePlus

    ... limited. Home Visit Global Sites Search Help? White Blood Cell Count Share this page: Was this page helpful? ... Count; Leukocyte Count; White Count Formal name: White Blood Cell Count Related tests: Complete Blood Count , Blood Smear , ...

  3. Cerebrospinal fluid white cell count: discriminatory or otherwise for enteroviral meningitis in infants and young children?

    PubMed

    Tan, Natalie Woon Hui; Lee, Elis Yuexian; Khoo, Gloria Mei Chin; Tee, Nancy Wen Sim; Krishnamoorthy, Subramania; Choong, Chew Thye

    2016-04-01

    Non-polio enteroviruses (EV) are the most common viruses causing aseptic meningitis in children. We aim to evaluate the cerebrospinal fluid (CSF) characteristics of neonates and children with EV meningitis with a view to determine whether it could be discriminatory or otherwise in making a positive diagnosis. We performed a 3-year (July 2008-July 2011) retrospective study of children ≤16 years, treated at a tertiary children's hospital, with positive CSF EV polymerase chain reaction (PCR) and negative blood and CSF bacterial cultures. A total of 206 children were studied. The median CSF white cell count was 79 cells/mm(3) (range 0-4608 cells/mm(3)). CSF pleocytosis was observed in 99/150 (66%) aged ≤90 days, 3/4 (75%) aged 90 days-1 year, and 49/52 (94%) children ≥3 years. There was a huge variability in CSF pleocytosis in infants ≤90 days, where 34% of them had no pleocytosis, while in 66%, a wide range of pleocytosis that might even suggest bacterial meningitis was noted. CSF red cells were low, and protein or sugar values were not discriminatory. CSF pleocytosis in relation to increasing age was found to be statistically significant (p < 0.001). Early lumbar puncture within 48 h of symptoms and absence of CSF pleocytosis was also statistically significant (p = 0.039). CSF pleocytosis in EV meningitis is commoner in older children. As there was a huge variability in CSF pleocytosis in infants ≤90 days particularly, CSF analysis including EV PCR could avoid unnecessary antibiotic therapy.

  4. 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.

  5. Interpretation of Amniotic Fluid White Blood Cell Count in “Bloody Tap” Amniocenteses in Women With Symptoms of Preterm Labor

    PubMed Central

    Abdel-Razeq, Sonya S.; Buhimschi, Irina A.; Bahtiyar, Mert O.; Rosenberg, Victor A.; Dulay, Antonette T.; Han, Christina S.; Werner, Erika F.; Thung, Stephen; Buhimschi, Catalin S.

    2014-01-01

    Objective To estimate whether blood-contaminated amniotic fluid affects the performance of white blood cell (WBC) count in diagnosing intraamniotic inflammation and infection. Methods Three hundred fifty-seven consecutive women pregnant with singletons undergoing amniocentesis to rule out infection were enrolled prospectively. A “bloody tap” was defined as a red blood cell (RBC) count of 1,000 cells/mm3 or more. Proteomics analysis of amniotic fluid was used in this study as the standard for diagnosing inflammation. Infection was confirmed by positive amniotic fluid culture. An amniotic fluid WBC count correction formula was computed using maternal WBC count, hematocrit, and mean corpuscular volume. Results The prevalence of a bloody tap amniocentesis was 22% (77 of 357). In the absence of inflammation, the amniotic fluid WBC count was significantly higher in bloody tap (median [interquartile range] 18 [9–58] cells/mm3) compared with non–bloody tap specimens (4 [1–10] cells/mm3; P<.001). The correction formula reversed this difference to a nonsignificant level (bloody tap 0 [0–17] compared with non–bloody tap 3 [1–10] cells/mm3; P=.273). In the setting of inflammation, the observed WBC count of bloody tap samples (778 [197–2,062 cells/mm3]) was significantly elevated compared with that of the non–bloody tap specimens (616 [105–1,730] cells/mm3; P=.023). Correction of the WBC count in bloody tap amniocenteses improved the test accuracy and positive likelihood ratios for inflammation and infection. A correction algorithm was not useful in amniotic fluid specimens with less than 1,000/RBCs/mm3 or WBC counts more than 1,100 cells/mm3. Given the nonlinear relationship between amniotic fluid WBC and RBC, for a rapid correction of WBC count, the number of neutrophils that need to be subtracted from the observed WBC count is variable. Conclusion In the setting of an amniotic fluid sample contaminated with 1,000 RBCs/mm3 or more, WBC count is a less

  6. High Red Blood Cell Count

    MedlinePlus

    Symptoms High red blood cell count By Mayo Clinic Staff A high red blood cell count is an increase in oxygen-carrying cells in your bloodstream. Red blood cells transport oxygen from your lungs to tissues throughout ...

  7. Low White Blood Cell Count

    MedlinePlus

    Symptoms Low white blood cell count By Mayo Clinic Staff A low white blood cell count (leukopenia) is a decrease in disease-fighting cells ( ... a decrease in a certain type of white blood cell (neutrophil). The definition of low white blood cell ...

  8. Low white blood cell count and cancer

    MedlinePlus

    Neutropenia and cancer; Absolute neutrophil count and cancer; ANC and cancer ... A person with cancer can get a low white blood cell count from the cancer or from treatment for the cancer. Cancer may ...

  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.

  10. Single Entity Electrochemistry Progresses to Cell Counting.

    PubMed

    Gooding, J Justin

    2016-10-10

    Red blood cells have been counted in an electrochemical collision experiment recently described by Compton and co-workers. As a cell collides with the electrode it lyses and a current is observed from the reduction of oxygen from within the cell.

  11. 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 μL 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.

  12. 21 CFR 864.6160 - Manual blood cell counting device.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 21 Food and Drugs 8 2013-04-01 2013-04-01 false Manual blood cell counting device. 864.6160... blood cell counting device. (a) Identification. A manual blood cell counting device is a device used to count red blood cells, white blood cells, or blood platelets. (b) Classification. Class I...

  13. 21 CFR 864.6160 - Manual blood cell counting device.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 21 Food and Drugs 8 2014-04-01 2014-04-01 false Manual blood cell counting device. 864.6160... blood cell counting device. (a) Identification. A manual blood cell counting device is a device used to count red blood cells, white blood cells, or blood platelets. (b) Classification. Class I...

  14. 21 CFR 864.6160 - Manual blood cell counting device.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 21 Food and Drugs 8 2012-04-01 2012-04-01 false Manual blood cell counting device. 864.6160... blood cell counting device. (a) Identification. A manual blood cell counting device is a device used to count red blood cells, white blood cells, or blood platelets. (b) Classification. Class I...

  15. 21 CFR 864.6160 - Manual blood cell counting device.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Manual blood cell counting device. 864.6160... blood cell counting device. (a) Identification. A manual blood cell counting device is a device used to count red blood cells, white blood cells, or blood platelets. (b) Classification. Class I...

  16. 21 CFR 864.6160 - Manual blood cell counting device.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 21 Food and Drugs 8 2011-04-01 2011-04-01 false Manual blood cell counting device. 864.6160... blood cell counting device. (a) Identification. A manual blood cell counting device is a device used to count red blood cells, white blood cells, or blood platelets. (b) Classification. Class I...

  17. 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.

  18. Optical planar waveguide for cell counting

    NASA Astrophysics Data System (ADS)

    LeBlanc, John; Mueller, Andrew J.; Prinz, Adrian; Butte, Manish J.

    2012-01-01

    Low cost counting of cells has medical applications in screening, military medicine, disaster medicine, and rural healthcare. In this report, we present a shallow, buried, planar waveguide fabricated by potassium ion exchange in glass that enables low-cost and rapid counting of metal-tagged objects that lie in the evanescent field of the waveguide. Laser light transmitted through the waveguide was attenuated proportionately to the presence of metal-coated microstructures fabricated from photoresist. This technology enables the low-cost enumeration of cells from blood, urine, or other biofluids.

  19. Counting white blood cells using morphological granulometries

    NASA Astrophysics Data System (ADS)

    Theera-Umpon, Nipon; Gader, Paul D.

    2000-04-01

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

  20. Effect of saliva contamination on induced sputum cell counts, IL-8 and eosinophil cationic protein levels.

    PubMed

    Simpson, J L; Timmins, N L; Fakes, K; Talbot, P I; Gibson, P G

    2004-05-01

    Excessive salivary contamination of induced sputum samples prevents the satisfactory examination of lower airway inflammation. The effects of salivary contamination on different sputum fluid phase measures and the levels of salivary contamination preventing analysis are not defined. The present study sought to examine this by investigating the effect of increasing salivary contamination on induced sputum samples. Sputum and saliva samples from subjects with asthma and healthy controls were collected, and treated with dithiothreitol (DTT). Saliva was then added to aliquots of dispersed sputum in increasing proportions (0% to 100%). The effect of increasing saliva contamination was assessed on sputum total cell count, viability, differential cell count and fluid phase levels of interleukin (IL)-8, eosinophil cationic protein (ECP) and total protein. The addition of saliva to induced sputum reduced total cell counts and absolute cell counts but did not change the differential cell count. Levels of fluid phase ECP and IL-8 were significantly reduced with increased salivary contamination. There was a progressive reduction in ECP and IL-8, which reached significance at 70% and 80% saliva contamination, respectively. IL-8 levels corrected for total protein showed no change with increasing saliva concentrations. Induced sputum differential cell counts expressed as the proportion of nonsquamous cells are robust measures that are not influenced by salivary contamination. Studies reporting total and absolute cell counts and fluid phase mediator levels require control for squamous contamination.

  1. Smart fast blood counting of trace volumes of body fluids from various mammalian species using a compact custom-built microscope cytometer (Conference Presentation)

    NASA Astrophysics Data System (ADS)

    Smith, Zachary J.; Gao, Tingjuan; Lin, Tzu-Yin; Carrade-Holt, Danielle; Lane, Stephen M.; Matthews, Dennis L.; Dwyre, Denis M.; Wachsmann-Hogiu, Sebastian

    2016-03-01

    Cell counting in human body fluids such as blood, urine, and CSF is a critical step in the diagnostic process for many diseases. Current automated methods for cell counting are based on flow cytometry systems. However, these automated methods are bulky, costly, require significant user expertise, and are not well suited to counting cells in fluids other than blood. Therefore, their use is limited to large central laboratories that process enough volume of blood to recoup the significant capital investment these instruments require. We present in this talk a combination of a (1) low-cost microscope system, (2) simple sample preparation method, and (3) fully automated analysis designed for providing cell counts in blood and body fluids. We show results on both humans and companion and farm animals, showing that accurate red cell, white cell, and platelet counts, as well as hemoglobin concentration, can be accurately obtained in blood, as well as a 3-part white cell differential in human samples. We can also accurately count red and white cells in body fluids with a limit of detection ~3 orders of magnitude smaller than current automated instruments. This method uses less than 1 microliter of blood, and less than 5 microliters of body fluids to make its measurements, making it highly compatible with finger-stick style collections, as well as appropriate for small animals such as laboratory mice where larger volume blood collections are dangerous to the animal's health.

  2. Photon Counts Statistics in Leukocyte Cell Dynamics

    NASA Astrophysics Data System (ADS)

    van Wijk, Eduard; van der Greef, Jan; van Wijk, Roeland

    2011-12-01

    In the present experiment ultra-weak photon emission/ chemiluminescence from isolated neutrophils was recorded. It is associated with the production of reactive oxygen species (ROS) in the "respiratory burst" process which can be activated by PMA (Phorbol 12-Myristate 13-Acetate). Commonly, the reaction is demonstrated utilizing the enhancer luminol. However, with the use of highly sensitive photomultiplier equipment it is also recorded without enhancer. In that case, it can be hypothesized that photon count statistics may assist in understanding the underlying metabolic activity and cooperation of these cells. To study this hypothesis leukocytes were stimulated with PMA and increased photon signals were recorded in the quasi stable period utilizing Fano factor analysis at different window sizes. The Fano factor is defined by the variance over the mean of the number of photon within the observation time. The analysis demonstrated that the Fano factor of true signal and not of the surrogate signals obtained by random shuffling increases when the window size increased. It is concluded that photon count statistics, in particular Fano factor analysis, provides information regarding leukocyte interactions. It opens the perspective to utilize this analytical procedure in (in vivo) inflammation research. However, this needs further validation.

  3. Inflammatory Cytokines and White Blood Cell Counts ...

    EPA Pesticide Factsheets

    Epidemiological observations of urban inhalation exposures to diesel exhaust (DE) and ozone (O3) have shown pre-clinical cardiopulmonary responses in humans. Identifying the key biological mechanisms that initiate these health bioindicators is difficult due to variability in environmental exposure in time and from person to person. Previously, environmentally controlled human exposure chambers have been used to study DE and O3 dose-response patterns separately, but investigation of co-exposures has not been performed under controlled conditions. Because a mixture is a more realistic exposure scenario for the general public, in this study we investigate the relationships of urban levels of urban-level DE exposure (300 μg/m3), O3 (0.3 ppm), DE + O3 co-exposure, and innate immune system responses. Fifteen healthy human volunteers were studied for changes in ten inflammatory cytokines (interleukins 1β, 2, 4, 5, 8, 10, 12p70 and 13, IFN-γ, and TNF-α) and counts of three white blood cell types (lymphocytes, monocytes, and neutrophils) following controlled exposures to DE, O3, and DE+O3. The results show subtle cytokines responses to the diesel-only and ozone-only exposures, and that a more complex (possibly synergistic) relationship exists in the combination of these two exposures with suppression of IL-5, IL-12p70, IFN-γ, and TNF-α that persists up to 22-hours for IFN-γ and TNF-α. The white blood cell differential counts showed significant monocyte and lympho

  4. Method of detecting and counting bacteria in body fluids

    NASA Technical Reports Server (NTRS)

    Chappelle, E. W.; Picciolo, G. L. (Inventor)

    1973-01-01

    A novel method is reported for determining bacterial levels in urine samples, which method depends on the quantitative determination of bacterial adenosine triphosphate (ATP) in the presence of non-bacterial ATP. After the removal of non-bacterial ATP, the bacterial ATP is released by cell rupture and is measured by an enzymatic bioluminescent assay using an enzyme obtained from the firefly.

  5. Amniotic fluid LPCAT1 mRNA correlates with the lamellar body count.

    PubMed

    Welch, Robert A; Shaw, Michael K; Welch, Kathryn C

    2016-07-01

    Lysophosphatidylcholine acyltransferase 1 (LPCAT1) is required in the biosynthesis of pulmonary surfactant. This short communication describes our assessment of LPCAT1 mRNA levels in human amniotic fluid. We found a direct correlation between LPCAT1 mRNA copies and the amniotic fluid lamellar body count (LBC). This finding corroborates an association between LPCAT1 and surfactant phospholipid biosynthesis in humans. It may provide a model for future research in perinatal medicine.

  6. Polymorphonuclear counts in ascitic fluid and microorganisms producing spontaneous bacterial peritonitis: an under-recognized relationship.

    PubMed

    Ariza, Xavier; Lora-Tamayo, Jaime; Castellote, José; Xiol, Xavier; Ariza, Javier

    2013-10-01

    BACKGROUND AND AIMS. In cirrhotic patients with spontaneous bacterial peritonitis (SBP) higher polymorphonuclear (PMN) count in ascitic fluid have been reported in infections caused by Gram-negative bacilli (GNB) as opposed to Gram-positive cocci (GPC). However, the influence of other associated factors on the PMN count, such as the specific microorganism causing the episode of SBP, has not been well established. METHODS. Retrospective observational study of 194 episodes of positive ascitic and/or blood culture SBP in 159 patients with liver cirrhosis (2001-2009). Parameters associated with PMN count in ascitic fluid at diagnosis were evaluated. RESULTS. The multivariate analysis (model 1) showed that a virulent etiology of the infection [coefficient 3.941 (95% confidence interval (95 CI): 0.421-7.461)] and the model for end-stage liver disease (MELD) score [coefficient 0.196 (95 CI: 0.007-0.384)] were positively associated with the PMN count in ascites, while a nosocomial acquisition was inversely associated [coefficient -3.546 (95 CI: -6.855 - -0.238)]. A nonsignificant trend toward higher PMN count was found in GNB versus GPC, but there were differences between groups of microorganisms: pyogenic streptococci [median (p25-p75): 3211 (1615-8004)], Enterobacteriaceae [2958 (917-7690)], Vibrionaceae [9215 (375-17280)], nonfermenting GNB [1384 (565-3865)], viridans group streptococci [1044 (503-2354)] and enterococci [1050 (476-4655)](p = 0.005). No clear cut-offs of ascitic PMN count predicting a particular etiology could be calculated out of these data. CONCLUSIONS. In cirrhotic patients with SBP, the causing microorganism, the place of acquisition of the infection and the host liver condition were the main factors determining PMN count in ascitic fluid. Third-generation cephalosporin resistance was associated with low PMN count probably because this group included bacteria with inherent low virulence.

  7. Geophysical Fluid Flow Cell Simulation

    NASA Technical Reports Server (NTRS)

    1998-01-01

    Computer simulation of atmospheric flow corresponds well to imges taken during the second Geophysical Fluid Flow Cell (BFFC) mission. The top shows a view from the pole, while the bottom shows a view from the equator. Red corresponds to hot fluid rising while blue shows cold fluid falling. This simulation was developed by Anil Deane of the University of Maryland, College Park and Paul Fischer of Argorne National Laboratory. Credit: NASA/Goddard Space Flight Center

  8. 21 CFR 864.8185 - Calibrator for red cell and white cell counting.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 21 Food and Drugs 8 2013-04-01 2013-04-01 false Calibrator for red cell and white cell counting... Calibrator for red cell and white cell counting. (a) Identification. A calibrator for red cell and white cell counting is a device that resembles red or white blood cells and that is used to set instruments...

  9. 21 CFR 864.8185 - Calibrator for red cell and white cell counting.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 21 Food and Drugs 8 2012-04-01 2012-04-01 false Calibrator for red cell and white cell counting... Calibrator for red cell and white cell counting. (a) Identification. A calibrator for red cell and white cell counting is a device that resembles red or white blood cells and that is used to set instruments...

  10. 21 CFR 864.8185 - Calibrator for red cell and white cell counting.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 21 Food and Drugs 8 2014-04-01 2014-04-01 false Calibrator for red cell and white cell counting... Calibrator for red cell and white cell counting. (a) Identification. A calibrator for red cell and white cell counting is a device that resembles red or white blood cells and that is used to set instruments...

  11. 21 CFR 864.8185 - Calibrator for red cell and white cell counting.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 21 Food and Drugs 8 2011-04-01 2011-04-01 false Calibrator for red cell and white cell counting... Calibrator for red cell and white cell counting. (a) Identification. A calibrator for red cell and white cell counting is a device that resembles red or white blood cells and that is used to set instruments...

  12. 21 CFR 864.8185 - Calibrator for red cell and white cell counting.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Calibrator for red cell and white cell counting... Calibrator for red cell and white cell counting. (a) Identification. A calibrator for red cell and white cell counting is a device that resembles red or white blood cells and that is used to set instruments...

  13. Differential leucocyte count for ewe milk with low and high somatic cell count.

    PubMed

    Albenzio, Marzia; Caroprese, Mariangela

    2011-02-01

    This study was undertaken to compare flow cytometry (FC) and direct microscopic leucocyte count (MDLC) for the differentiation of macrophages, lymphocytes and polymorphonuclear leucocyte (PMN) and to evaluate leucocyte distribution in ewe milk with low and high somatic cell count (SCC). Milk samples were grouped for somatic cell count in low SCC (LSCC) when the content was lower than 5·00 × 10(5)/ml and high SCC (HSCC) when the content was higher than 1·00 × 10(6)/ml. No differences were found between the two methods tested suggesting that FC could be used as a routine test for rapid discrimination of leucocytes. Percentages of lymphocytes in ewe milk were higher in LSCC (50%) than in HSCC (39%) and count ranged from 273·91 ± 56·62 × 10(3) cells/ml (LSCC) to 308·90 ± 46·15 × 10(3) cells/ml (HSCC). PMN number was lower in LSCC than in HSCC (248·83 ± 46·87 × 10(3) cells/ml v. 444·38 ± 58·62 × 10(3) cells/ml); accordingly the percentage was lower in LSCC (40%) than in HSCC (57%). No differences were found for macrophages which were 36·36 ± 5·51 × 10(3) cells/ml and 39·32 ± 6·83 × 10(3) cells/ml in LSCC and HSCC, respectively. Lymphocytes in ewe milk did not vary with increased number of somatic cells and were the predominant cell type in LSCC. PMN represented the main population detected in HSCC and the correlation with SCC evidenced that this leucocyte class could be useful in differentiating ewe milk cell count, being strictly responsible for the SCC increase.

  14. Peripheral blood CD34+ cell count reliably predicts autograft yield.

    PubMed

    Chapple, P; Prince, H M; Quinn, M; Bertoncello, I; Juneja, S; Wolf, M; Januszewicz, H; Brettell, M; Gardyn, J; Seymour, C; Venter, D

    1998-07-01

    A reliable measure to predict peripheral blood progenitor cell (PBPC) autograft CD34+ cell content is required to optimize the timing of PBPC collection. We prospectively examined the peripheral blood (PB) CD34+ cell count in 59 consecutive patients with various malignancies and analyzed the correlation between the PB CD34+ cell count and various parameters in the PBPC autograft. Two hundred and thirty-five collections were performed with a median of 4.0 collections per patient (range, 2-10). The median PB CD34+ cell count at the time of collection was 39 x 10(6)/1 (range, 0.0-285.6). The PBPC autograft parameters measured were the CD34+ cell, colony-forming unit granulocyte-macrophage (CFU-GM) and mononuclear cell (MNC) content. There was a strong linear correlation between PB CD34+ cells/l and autograft CD34+ cells/kg (r = 0.8477). The correlation with CFU-GM/kg (r = 0.5512) was weaker. There was no correlation between autograft CD34+ cells/kg and PB WBC (r= 0.0684), PB MNC (r = 0.1518) or PB platelet count (r = 0.2010). At our institution we aim to obtain a minimum of 0.5 x 10(6) CD34+ cells/kg with each day of collection. We demonstrate that such a collection can be reliably obtained if the PB CD34+ cell count exceeds 5.0 x 10(6)/l.

  15. HIV-1 outcompetes HIV-2 in dually infected Senegalese individuals with low CD4+ cell counts

    PubMed Central

    Raugi, Dana N.; Gottlieb, Geoffrey S.; Sow, Papa S.; Toure, Macoumba; Sall, Fatima; Gaye, Awa; N’doye, Ibra; Kiviat, Nancy B.; Hawes, Stephen E.

    2014-01-01

    Objective Dual infection with HIV-1 and HIV-2, which is not uncommon in West Africa, has implications for transmission, progression, and antiretroviral therapy (ART). Few studies have examined viral dynamics in this setting. Our objective was to directly compare HIV-1 and HIV-2 viral loads and to examine whether this relationship is associated with CD4+ cell count. Study design This is a retrospective analysis of data from observational cohort studies. Methods We compared HIV-1 and HIV-2 viral loads from 65 dually infected, ART-naive Senegalese individuals. Participants provided blood, oral fluid, and cervicovaginal lavage (CVL) or semen samples for virologic and immunologic testing. We assessed relationships between HIV-1 and HIV-2 levels using linear regression with generalized estimating equations to account for multiple study visits. Results After adjusting for CD4+ cell count, age, sex, and commercial sex work, HIV-1 RNA levels were significantly higher than HIV-2 levels in semen, CVL, and oral fluids. Despite similar peripheral blood mononuclear cell DNA levels among individuals with CD4+ cell counts above 500 cells/µl, individuals with CD4+ cell counts below 500 cells/µl had higher HIV-1 and lower HIV-2 DNA levels. Individuals with high CD4+ cell counts had higher mean HIV-1 plasma RNA viral loads than HIV-2, with HIV-1 levels significantly higher and HIV-2 levels trending toward lower mean viral loads among individuals with low CD4+ cell counts. Conclusion Our data are consistent with the hypothesis that with disease progression, HIV-1 outcompetes HIV-2 in dually infected individuals. This finding helps explain differences in prevalence and outcomes between HIV-1, HIV-2, and HIV-dual infection. PMID:23665777

  16. Mast cell and histamine content of human bronchoalveolar lavage fluid.

    PubMed Central

    Agius, R M; Godfrey, R C; Holgate, S T

    1985-01-01

    Bronchoalveolar lavage was performed in 97 patients including control patients with bronchial carcinoma (24) and patients with sarcoidosis (20), cryptogenic fibrosing alveolitis (9), and asthma (4), and others. Cytocentrifuged slides were stained by two methods: May-Grünwald Giemsa and toluidine blue. In the last 32 subjects the bronchoalveolar lavage fluid was separated into supernatant and cell pellet for the subsequent assay of the performed mast cell mediator, histamine. Comparison of the two methods of staining showed a bias towards toluidine blue. Controls had a differential mean (SE) mast cell count of 0.07% (0.01%). Higher counts were noted in cryptogenic fibrosing alveolitis--0.61% (0.15%) (p less than 0.001)--and in sarcoidosis--0.14% (0.02%) (p less than 0.05). There was a strong correlation between absolute mast cell counts and cell lysate histamine concentration (r = 0.78, p less than 0.001). Less strong, significant, correlations between supernatant histamine concentration and absolute mast cell counts (r = 0.48, p less than 0.01) or cell lysate histamine concentration (r = 0.72, p less than 0.01) were also found. Derived mean values of histamine per mast cell ranged from 3.7 to 10.9 picograms. The mean histamine content of lavage fluid supernatant as a percentage of the total lavage fluid histamine was 24.9% (3.3%). The possible clinical significance of these findings is discussed. Images PMID:4060097

  17. 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.

  18. [Automated hematology analysers and spurious counts Part 3. Haemoglobin, red blood cells, cell count and indices, reticulocytes].

    PubMed

    Godon, Alban; Genevieve, Franck; Marteau-Tessier, Anne; Zandecki, Marc

    2012-01-01

    Several situations lead to abnormal haemoglobin measurement or to abnormal red blood cells (RBC) counts, including hyperlipemias, agglutinins and cryoglobulins, haemolysis, or elevated white blood cells (WBC) counts. Mean (red) cell volume may be also subject to spurious determination, because of agglutinins (mainly cold), high blood glucose level, natremia, anticoagulants in excess and at times technological considerations. Abnormality related to one measured parameter eventually leads to abnormal calculated RBC indices: mean cell haemoglobin content is certainly the most important RBC parameter to consider, maybe as important as flags generated by the haematology analysers (HA) themselves. In many circumstances, several of the measured parameters from cell blood counts (CBC) may be altered, and the discovery of a spurious change on one parameter frequently means that the validity of other parameters should be considered. Sensitive flags allow now the identification of several spurious counts, but only the most sophisticated HA have optimal flagging, and simpler ones, especially those without any WBC differential scattergram, do not share the same capacity to detect abnormal results. Reticulocytes are integrated into the CBC in many HA, and several situations may lead to abnormal counts, including abnormal gating, interference with intraerythrocytic particles, erythroblastosis or high WBC counts.

  19. Development of a stained cell nuclei counting system

    NASA Astrophysics Data System (ADS)

    Timilsina, Niranjan; Moffatt, Christopher; Okada, Kazunori

    2011-03-01

    This paper presents a novel cell counting system which exploits the Fast Radial Symmetry Transformation (FRST) algorithm [1]. The driving force behind our system is a research on neurogenesis in the intact nervous system of Manduca Sexta or the Tobacco Hornworm, which was being studied to assess the impact of age, food and environment on neurogenesis. The varying thickness of the intact nervous system in this species often yields images with inhomogeneous background and inconsistencies such as varying illumination, variable contrast, and irregular cell size. For automated counting, such inhomogeneity and inconsistencies must be addressed, which no existing work has done successfully. Thus, our goal is to devise a new cell counting algorithm for the images with non-uniform background. Our solution adapts FRST: a computer vision algorithm which is designed to detect points of interest on circular regions such as human eyes. This algorithm enhances the occurrences of the stained-cell nuclei in 2D digital images and negates the problems caused by their inhomogeneity. Besides FRST, our algorithm employs standard image processing methods, such as mathematical morphology and connected component analysis. We have evaluated the developed cell counting system with fourteen digital images of Tobacco Hornworm's nervous system collected for this study with ground-truth cell counts by biology experts. Experimental results show that our system has a minimum error of 1.41% and mean error of 16.68% which is at least forty-four percent better than the algorithm without FRST.

  20. Single proton counting at the RIKEN cell irradiation facility

    SciTech Connect

    Mäckel, V. Puttaraksa, N.; Kobayashi, T.; Yamazaki, Y.

    2015-08-15

    We present newly developed tapered capillaries with a scintillator window, which enable us to count single protons at the RIKEN cell irradiation setup. Their potential for performing single proton irradiation experiments at our beamline setup is demonstrated with CR39 samples, showing a single proton detection fidelity of 98%.

  1. Fluorescent Cell-Counting Neutralization Test for Psittacosis

    PubMed Central

    Hahon, Nicholas; Cooke, Kenneth O.

    1965-01-01

    Hahon, Nicholas (Fort Detrick, Frederick, Md.), and Kenneth O. Cooke. Fluorescent cell-counting neutralization test for psittacosis. J. Bacteriol. 89:1465–1471. 1965.—A sensitive, precise, and specific serological procedure, the fluorescent cell-counting neutralization test, was developed to detect and to measure quantitatively psittacosis serum-neutralizing antibodies within 24 hr. The test is based on the reduction of fluorescent cells in McCoy cell monolayers resulting from the neutralization of infective agent particles by specific antiserum. Small but significant rises in neutralizing titers were measured in serum specimens from monkeys previously exposed to the psittacosis agent and from humans with diagnoses of subclinical or established psittacosis infections. PMID:14291581

  2. Automatic counting of microglial cell activation and its applications

    PubMed Central

    Gallego, Beatriz I.; de Gracia, Pablo

    2016-01-01

    Glaucoma is a multifactorial optic neuropathy characterized by the damage and death of the retinal ganglion cells. This disease results in vision loss and blindness. Any vision loss resulting from the disease cannot be restored and nowadays there is no available cure for glaucoma; however an early detection and treatment, could offer neuronal protection and avoid later serious damages to the visual function. A full understanding of the etiology of the disease will still require the contribution of many scientific efforts. Glial activation has been observed in glaucoma, being microglial proliferation a hallmark in this neurodegenerative disease. A typical project studying these cellular changes involved in glaucoma often needs thousands of images - from several animals - covering different layers and regions of the retina. The gold standard to evaluate them is the manual count. This method requires a large amount of time from specialized personnel. It is a tedious process and prone to human error. We present here a new method to count microglial cells by using a computer algorithm. It counts in one hour the same number of images that a researcher counts in four weeks, with no loss of reliability. PMID:27651757

  3. A New Computer Controlled Blood Cell Counting and Sizing System

    PubMed Central

    Talug, E.; Arp, L. J.

    1977-01-01

    A new automated particle counting and sizing system was used in this investigation. The Electrozone/Celloscope® counter electronically detects the passage of red blood cells in a saline suspension. It was used to automatically plot the size distribution curves for graphically displaying the effects of sequentially increased hemolysis levels in blood samples. In this study the size distributions and counts of red blood cells in a saline solution of various concentrations (0.85%, 0.75%, 0.65%, 0.60%, 0.55%, 0.50%, 0.45%, 0.40%, 0.35%, 0.30%, 0.20%, 0.10%) were analyzed and compared to normal values. The accuracy and reproducibility of counts with the electronic counter are compared to hemocytometer counts. Some of the advantages and disadvantages of both systems are discussed. Normal size distribution curves first shift slightly to a larger particle size due to osmotic swelling (in 0.65%, 0.60%, 0.55% of saline solution). Upon further reduction of the saline concentration to .45%, .40%, .35%, hemolysis is observed as generation of the second peak showing clearly the smaller particle size.

  4. Flow cytometric bacterial cell counts challenge conventional heterotrophic plate counts for routine microbiological drinking water monitoring.

    PubMed

    Van Nevel, S; Koetzsch, S; Proctor, C R; Besmer, M D; Prest, E I; Vrouwenvelder, J S; Knezev, A; Boon, N; Hammes, F

    2017-04-15

    Drinking water utilities and researchers continue to rely on the century-old heterotrophic plate counts (HPC) method for routine assessment of general microbiological water quality. Bacterial cell counting with flow cytometry (FCM) is one of a number of alternative methods that challenge this status quo and provide an opportunity for improved water quality monitoring. After more than a decade of application in drinking water research, FCM methodology is optimised and established for routine application, supported by a considerable amount of data from multiple full-scale studies. Bacterial cell concentrations obtained by FCM enable quantification of the entire bacterial community instead of the minute fraction of cultivable bacteria detected with HPC (typically < 1% of all bacteria). FCM measurements are reproducible with relative standard deviations below 3% and can be available within 15 min of samples arriving in the laboratory. High throughput sample processing and complete automation are feasible and FCM analysis is arguably less expensive than HPC when measuring more than 15 water samples per day, depending on the laboratory and selected staining procedure(s). Moreover, many studies have shown FCM total (TCC) and intact (ICC) cell concentrations to be reliable and robust process variables, responsive to changes in the bacterial abundance and relevant for characterising and monitoring drinking water treatment and distribution systems. The purpose of this critical review is to initiate a constructive discussion on whether FCM could replace HPC in routine water quality monitoring. We argue that FCM provides a faster, more descriptive and more representative quantification of bacterial abundance in drinking water.

  5. Relationship between somatic cell count, polymorphonuclear leucocyte count and quality parameters in bovine bulk tank milk.

    PubMed

    Wickström, Erik; Persson-Waller, Karin; Lindmark-Månsson, Helena; Ostensson, Karin; Sternesjö, Ase

    2009-05-01

    The somatic cell count (SCC) in bovine bulk tank milk is presently used as an indicator of raw milk quality, reflecting the udder health status of the herd. During mastitis, SCC increases, mostly owing to an influx of polymorphonuclear leucocytes (PMN) from blood into milk, with a concomitant change in milk composition. Bulk tank milk samples were categorized according to their SCC, as well as polymorphonuclear leucocyte count (PMNC), to study relationships between SCC, PMNC and various raw milk quality traits, i.e. contents of total protein, whey protein, casein, fat and lactose, casein number, proteolysis and rheological properties. The proportion of PMN, obtained by direct microscopy, was significantly higher in samples with high SCC compared with low SCC samples. SCC and PMNC were strongly correlated, yielding a correlation coefficient of 0.85. High SCC samples had lower lactose and casein contents, lower casein number and more proteolysis than low SCC samples. Samples with high PMNC had a lower casein number than low PMNC samples. Samples with high and low SCC or PMNC did not differ in respect to rheological properties. Our results do not indicate that PMNC is a better biomarker than SCC for raw bulk tank milk quality, as previously proposed.

  6. 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.

  7. Bone Density Loss Is Associated With Blood Cell Counts.

    PubMed

    Valderrábano, Rodrigo J; Lui, Li-Yung; Lee, Jennifer; Cummings, Steven R; Orwoll, Eric S; Hoffman, Andrew R; Wu, Joy Y

    2017-02-01

    Hematopoiesis depends on a supportive microenvironment. Preclinical studies in mice have demonstrated that osteoblasts influence the development of blood cells, particularly erythrocytes, B lymphocytes, and neutrophils. However, it is unknown whether osteoblast numbers or function impact blood cell counts in humans. We tested the hypothesis that men with low BMD or greater BMD loss have decreased circulating erythrocytes and lymphocytes and increased myeloid cells. We performed a cross-sectional analysis and prospective analysis in the Osteoporotic Fractures in Men (MrOS) study, a multisite longitudinal cohort study. A total of 2571 community-dwelling men (≥65 years) who were able to walk without assistance, did not have a hip replacement or fracture, and had complete blood counts (CBCs) at the third study visit were analyzed. Multivariable (MV)-adjusted logistic regression estimated odds of white blood cell (WBC) subtypes (highest and lowest quintile versus middle), and anemia (clinically defined) associated with BMD by DXA scan (at visit 3), annualized percent BMD change (baseline to visit 3), and high BMD loss (>0.5%/year, from baseline to visit 3) at the femoral neck (FN) and total hip (TH). MV-adjusted models included age, BMI, cancer history, smoking status, alcohol intake, corticosteroid use, self-reported health, thiazide use, and physical activity. At visit 3 greater TH BMD loss (per 1 SD) was associated with increased odds of anemia, high neutrophils, and low lymphocytes. Annualized BMD loss of >0.5% was associated with increased odds of anemia, high neutrophils, and low lymphocytes. Similar results were observed for FN BMD regarding anemia and lymphocytes. We conclude that community-dwelling older men with declining hip BMD over about 7 years had increased risks of anemia, lower lymphocyte count, and higher neutrophil count, consistent with preclinical studies. Bone health and hematopoiesis may have greater interdependency than previously recognized.

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

    PubMed

    Chiavaroli, Stefano; Newport, David; Woulfe, Bernie

    2010-06-15

    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 cellss is achieved.

  9. Digital cell counting device integrated with a single-cell array.

    PubMed

    Saeki, Tatsuya; Hosokawa, Masahito; Lim, Tae-kyu; Harada, Manabu; Matsunaga, Tadashi; Tanaka, Tsuyoshi

    2014-01-01

    In this paper, we present a novel cell counting method accomplished using a single-cell array fabricated on an image sensor, complementary metal oxide semiconductor sensor. The single-cell array was constructed using a microcavity array, which can trap up to 7,500 single cells on microcavities periodically arranged on a plane metallic substrate via the application of a negative pressure. The proposed method for cell counting is based on shadow imaging, which uses a light diffraction pattern generated by the microcavity array and trapped cells. Under illumination, the cell-occupied microcavities are visualized as shadow patterns in an image recorded by the complementary metal oxide semiconductor sensor due to light attenuation. The cell count is determined by enumerating the uniform shadow patterns created from one-on-one relationships with single cells trapped on the microcavities in digital format. In the experiment, all cell counting processes including entrapment of non-labeled HeLa cells from suspensions on the array and image acquisition of a wide-field-of-view of 30 mm(2) in 1/60 seconds were implemented in a single integrated device. As a result, the results from the digital cell counting had a linear relationship with those obtained from microscopic observation (r(2)  = 0.99). This platform could be used at extremely low cell concentrations, i.e., 25-15,000 cells/mL. Our proposed system provides a simple and rapid miniaturized cell counting device for routine laboratory use.

  10. Baseline correlation and comparative kinetics of cerebrospinal fluid colony-forming unit counts and antigen titers in cryptococcal meningitis.

    PubMed

    Brouwer, Annemarie E; Teparrukkul, Paprit; Pinpraphaporn, Supraphada; Larsen, Robert A; Chierakul, Wirongrong; Peacock, Sharon; Day, Nicholas; White, Nicholas J; Harrison, Thomas S

    2005-08-15

    Cerebrospinal fluid (CSF) cryptococcal colony-forming unit counts and CSF cryptococcal antigen titers serve as alternative measures of organism load in cryptococcal meningitis. For these measures, we correlated baseline values and rates of decline during the first 2 weeks of therapy in 68 human immunodeficiency virus--seropositive patients with cryptococcal meningitis. At baseline, there was a strong correlation between CSF cryptococcal colony-forming unit counts and CSF cryptococcal antigen titers. During the first 2 weeks of therapy, CSF cryptococcal colony-forming unit counts decreased by >5 logs, and CSF cryptococcal antigen titers decreased by 1.5 dilutions. In individual patients, there was no correlation between the rate of decline in CSF cryptococcal colony-forming unit counts and that in CSF cryptococcal antigen titers.

  11. Polycystic ovary syndrome and the peripheral blood white cell count.

    PubMed

    Herlihy, A C; Kelly, R E; Hogan, J L; O'Connor, N; Farah, N; Turner, M J

    2011-01-01

    This retrospective cross-sectional study examined if the white cell count (WCC) is increased in women with polycystic ovary syndrome (PCOS) and if so, is it due to PCOS or to the associated obesity? Body mass index (BMI) was calculated and body composition was measured using bioelectrical impedance analysis. Of the 113 women studied, 36 had PCOS and 77 did not. The mean WCC was higher in the PCOS group compared with the non-PCOS group (8.9 × 10(9)/l vs 7.4 × 10(9)/l p = 0.002). This increase was due to a higher neutrophil count (5.6 × 10(9)/l vs 4.3 × 10(9)/l; p = 0.003). There was a leucocytosis (WCC >11 × 10(9)/l) present in 19% of the PCOS group compared with 1% in the non-PCOS group (p < 0.001). The neutrophil count was abnormally high (>7.7 × 10(9)/l) in 14% of the PCOS group compared with 4% in the non-PCOS group (p < 0.001). On regression analysis, however, the only independent variable which explained both the increased WCC and the increased neutrophil count was PCOS. We found that PCOS is associated with an increased WCC due to increased neutrophils, which supports the evidence that PCOS is associated with low-grade inflammation. The increase appears to be due to the underlying PCOS, and not to the increased adiposity associated with PCOS.

  12. WBC count

    MedlinePlus

    Leukocyte count; White blood cell count ... blood is 4,500 to 11,000 white blood cells per microliter (mcL) or 4.5 to 11. ... LOW WHITE BLOOD CELL (WBC) COUNT A low number of WBCs is called leukopenia. A WBC count below 4500 is below normal ...

  13. White blood cell counting on smartphone paper electrochemical sensor.

    PubMed

    Wang, Xinhao; Lin, Guohong; Cui, Guangzhe; Zhou, Xiangfei; Liu, Gang Logan

    2017-04-15

    White blood cell (WBC) analysis provides rich information in rapid diagnosis of acute bacterial and viral infections as well as chronic disease management. For patients with immune deficiency or leukemia WBC should be persistently monitored. Current WBC counting method relies on bulky instrument and trained personnel and is time consuming. Rapid, low-cost and portable solution is in highly demand for point of care test. Here we demonstrate a label-free smartphone based electrochemical WBC counting device on microporous paper with patterned gold microelectrodes. WBC separated from whole blood was trapped by the paper with microelectrodes. WBC trapped on the paper leads to the ion diffusion blockage on microelectrodes, therefore cell concentration is determined by peak current on the microelectrodes measured by a differential pulse voltammeter and the quantitative results are collected by a smartphone wirelessly within 1min. We are able to rapidly quantify WBC concentrations covering the common physiological and pathological range (200-20000μL(-1)) with only 10μL sample and high repeatability as low as 10% in CoV (Coefficient of Variation). The unique smartphone paper electrochemical sensor ensures fast cell quantification to achieve rapid and low-cost WBC analysis at the point-of-care under resource limited conditions.

  14. Architecture and inflammatory cell composition of the feline lung with special consideration of eosinophil counts.

    PubMed

    Shibly, S; Klang, A; Galler, A; Schwendenwein, I; Christian, M; Guija, A; Tichy, A; Hirt, R A

    2014-05-01

    An increase in the number of eosinophils in bronchoalveolar lavage fluid (BALF) is a hallmark of feline asthma; however, a wide range in the percentage of eosinophils in BALF has been documented in healthy cats. In this study, BALF and lung tissue were collected from 15 cats without respiratory disease, BALF was taken from 15 cats with asthma and lung tissue was collected from six different asthmatic cats. Total nucleated cell count (TNCC) and inflammatory cell percentages were measured in BALF and lung tissue was evaluated microscopically. Asthmatic cats had a significantly higher eosinophil count in lung tissue, but BALF TNCC did not differ significantly between groups. Cats without respiratory signs had significantly more numerous macrophages and lymphocytes in BALF than asthmatics, but significantly lower percentages of eosinophils (4.2 ± 7.8% versus 49.4 ± 20.6%, P <0.001). In healthy feline airways a BALF eosinophil percentage of <5% can be expected. Dominant microscopical findings in feline asthma include high eosinophil counts, airway remodelling and inflammation. There is good correlation between the findings in BALF and tissue in feline asthma.

  15. 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.

  16. 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...

  17. Electrolyte and Fluid Transport in Mesothelial Cells

    PubMed Central

    Ji, Hong-Long; Nie, Hong-Guang

    2008-01-01

    Mesothelial cells are specialized epithelial cells, which line the pleural, pericardial, and peritoneal cavities. Accumulating evidence suggests that the monolayer of mesothelial cells is permeable to electrolyte and fluid, and thereby govern both fluid secretion and re-absorption in the serosal cavities. Disorders in these salt and fluid transport systems may be fundamental in the pathogenesis of pleural effusion, pericardial effusion, and ascites. In this review, we discuss the location, physiological function, and regulation of active transport (Na+-K+-ATPase) systems, cation and anion channels (Na+, K+, Cl−, and Ca2+ channels), antiport (exchangers) systems, and symport (co-transporters) systems, and water channels (aquaporins). These secretive and absorptive pathways across mesothelial monolayer cells for electrolytes and fluid may provide pivotal therapeutical targets for novel clinical intervention in edematous diseases of serous cavities. PMID:19169368

  18. Automatic Biological Cell Counting Using a Modified Gradient Hough Transform.

    PubMed

    Denimal, Emmanuel; Marin, Ambroise; Guyot, Stéphane; Journaux, Ludovic; Molin, Paul

    2017-02-01

    We present a computational method for pseudo-circular object detection and quantitative characterization in digital images, using the gradient accumulation matrix as a basic tool. This Gradient Accumulation Transform (GAT) was first introduced in 1992 by Kierkegaard and recently used by Kaytanli & Valentine. In the present article, we modify the approach by using the phase coding studied by Cicconet, and by adding a "local contributor list" (LCL) as well as a "used contributor matrix" (UCM), which allow for accurate peak detection and exploitation. These changes help make the GAT algorithm a robust and precise method to automatically detect pseudo-circular objects in a microscopic image. We then present an application of the method to cell counting in microbiological images.

  19. High-resolution particle analysis--its application to platelet counting and suggestions for further application in blood cell analysis.

    PubMed

    Haynes, J L

    1980-01-01

    The characteristics of an instrument for high-resolution particle analysis in flow are discussed. It employs a combination of hydrodynamic focusing, fluid resistors, and electronic techniques to achieve precision and ease of use heretofore unobtainable in a moderate-cost clinical instrument. Its application to whole blood platelet counting is discussed, and suggestions are made for its possible application to a wide variety of blood cell measurements.

  20. Application of a non-hazardous vital dye for cell counting with automated cell counters.

    PubMed

    Kim, Soo In; Kim, Hyun Jeong; Lee, Ho-Jae; Lee, Kiwon; Hong, Dongpyo; Lim, Hyunchang; Cho, Keunchang; Jung, Neoncheol; Yi, Yong Weon

    2016-01-01

    Recent advances in automated cell counters enable us to count cells more easily with consistency. However, the wide use of the traditional vital dye trypan blue (TB) raises environmental and health concerns due to its potential teratogenic effects. To avoid this chemical hazard, it is of importance to introduce an alternative non-hazardous vital dye that is compatible with automated cell counters. Erythrosin B (EB) is a vital dye that is impermeable to biological membranes and is used as a food additive. Similarly to TB, EB stains only nonviable cells with disintegrated membranes. However, EB is less popular than TB and is seldom used with automated cell counters. We found that cell counting accuracy with EB was comparable to that with TB. EB was found to be an effective dye for accurate counting of cells with different viabilities across three different automated cell counters. In contrast to TB, EB was less toxic to cultured HL-60 cells during the cell counting process. These results indicate that replacing TB with EB for use with automated cell counters will significantly reduce the hazardous risk while producing comparable results.

  1. Fuel cell membrane hydration and fluid metering

    DOEpatents

    Jones, Daniel O.; Walsh, Michael M.

    2003-01-01

    A hydration system includes fuel cell fluid flow plate(s) and injection port(s). Each plate has flow channel(s) with respective inlet(s) for receiving respective portion(s) of a given stream of reactant fluid for a fuel cell. Each injection port injects a portion of liquid water directly into its respective flow channel. This serves to hydrate at least corresponding part(s) of a given membrane of the corresponding fuel cell(s). The hydration system may be augmented by a metering system including flow regulator(s). Each flow regulator meters an injecting at inlet(s) of each plate of respective portions of liquid into respective portion(s) of a given stream of fluid by corresponding injection port(s).

  2. Fuel cell membrane hydration and fluid metering

    DOEpatents

    Jones, Daniel O.; Walsh, Michael M.

    1999-01-01

    A hydration system includes fuel cell fluid flow plate(s) and injection port(s). Each plate has flow channel(s) with respective inlet(s) for receiving respective portion(s) of a given stream of reactant fluid for a fuel cell. Each injection port injects a portion of liquid water directly into its respective flow channel in order to mix its respective portion of liquid water with the corresponding portion of the stream. This serves to hydrate at least corresponding part(s) of a given membrane of the corresponding fuel cell(s). The hydration system may be augmented by a metering system including flow regulator(s). Each flow regulator meters an injecting at inlet(s) of each plate of respective portions of liquid into respective portion(s) of a given stream of fluid by corresponding injection port(s).

  3. Progesterone metabolism in cultured amniotic fluid cells.

    PubMed

    Beling, C G; Cederqvist, L L

    1978-01-01

    Amniotic fluid cells obtained by amnicentesis at 16-20 weeks' gestation were grown in culture until a confluent monolayer of cell had been formed. Radiolabeled pregnenolone, progesterone and 20 alpha-dihydroprogesterone were added to the cell cultures; steroid metabolites which formed after 24 and 48 hours of incubation were identified. Incubation of the cell cultures with pregnenolone-3H resulted in the formation of progesterone, 17alpha-progesterone and 20 alpha-dihydroprogesterone. A significant amount of progesterone was identified after incubating the cell cultures with 20 alpha-dihydroprogesterone. The results indicate that 3 beta-ol-dehydrogenase, 17 alpha-hydroxylase and 20 alpha-hydroxysteroid dehydrogenase enzymes are present in cultured amniotic fluid cells obtained at 16-20 weeks' gestation.

  4. Reticulocyte Count Test

    MedlinePlus

    ... Reticulocyte Count Related tests: Red Blood Cell Count ; Hemoglobin ; Hematocrit ; Complete Blood Count ; Blood Smear ; Erythropoietin ; Vitamin ... on a complete blood count (CBC) , RBC count , hemoglobin or hematocrit , to help determine the cause To ...

  5. Mini-FLOTAC for counting Toxoplasma gondii oocysts from cat feces--comparison with cell counting plates.

    PubMed

    Djokic, Vitomir; Blaga, Radu; Rinaldi, Laura; Le Roux, Delphine; Ducry, Tamara; Maurelli, Maria Paola; Perret, Catherine; Djurkovic Djakovic, Olgica; Cringoli, Giuseppe; Boireau, Pascal

    2014-12-01

    Oocysts of Toxoplasma gondii represent one of the most common environmental contaminants causing the zoonotic infection toxoplasmosis. The aim of the present study was to compare the Mini-FLOTAC device with traditional cell counting plates (Kova Slide) for the detection of T. gondii oocysts from feline feces. Two types of experiments were performed: (i) purified oocysts were counted in different dilutions and (ii) specific pathogen free T. gondii-negative cat feces was inoculated with numbers of purified oocysts and counting was performed directly from feces. Our analysis showed a thousand times higher sensitivity of Mini-FLOTAC (5 × 10(2) oocysts) compared to Kova Slide (5 × 10(5) oocysts). Also, when compared by McNemar's test, counting of the purified oocysts showed a higher sensitivity of Mini-FLOTAC compared to Kova Slide, for a dilution of 10(3) oocysts/ml (chi(2) = 6.1; P < 0.05). A better sensitivity was also found with Mini-FLOTAC in dilutions of 10(5) and 10(4) oocysts/ml, when counted from feces (chi(2) = 4.2 and 8.1, respectively, P < 0.05). Our results show that Mini-FLOTAC is more sensitive than traditional methods of T. gondii oocysts detection and quantification is more accurate. Furthermore, Mini-FLOTAC simplicity and cost effectiveness allow it to be used with light microscopes in any laboratory or field conditions. We therefore recommend its use for regular screening. Further studies are needed to validate Mini-FLOTAC for the detection of oocysts in soil and water samples in field conditions.

  6. Nucleated red blood cell count in term and preterm newborns: reference values at birth.

    PubMed

    Perrone, S; Vezzosi, P; Longini, M; Marzocchi, B; Tanganelli, D; Testa, M; Santilli, T; Buonocore, G

    2005-03-01

    The prognostic value of nucleated red blood cell count at birth in relation to neonatal outcome has been established. However, reference values were needed to usefully interpret this variable. The normal range of reference values for absolute nucleated red blood cell count in 695 preterm and term newborns is reported.

  7. Association of psychological stress response of fatigue with white blood cell count in male daytime workers.

    PubMed

    Nishitani, Naoko; Sakakibara, Hisataka

    2014-01-01

    Relationships between work-related psychological and physical stress responses and counts of white blood cells (WBCs), neutrophils, and lymphocytes were investigated in 101 daytime workers. Counts of WBCs and neutrophils were positively associated with smoking and inversely correlated with high density lipoprotein (HDL)-cholesterol levels. Additionally, general fatigue score as measured by the profile of mood state was positively correlated with WBC and neutrophil counts whereas lymphocyte counts was not significantly associated with fatigue score. Multiple regression analysis showed that WBC count was significantly related to general fatigue, age, and HDL-cholesterol levels. Neutrophil count was significantly related to HDL-cholesterol levels and fatigue score. Among various psychological stress response variables, general fatigue may be a key determinant of low-grade inflammation as represented by increases of WBC and neutrophil counts.

  8. 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.

  9. 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.

  10. Micro-a-fluidics ELISA for Rapid CD4 Cell Count at the Point-of-Care

    PubMed Central

    Wang, ShuQi; Tasoglu, Savas; Chen, Paul Z.; Chen, Michael; Akbas, Ragip; Wach, Sonya; Ozdemir, Cenk Ibrahim; Gurkan, Umut Atakan; Giguel, Francoise F.; Kuritzkes, Daniel R.; Demirci, Utkan

    2014-01-01

    HIV has become one of the most devastating pathogens in human history. Despite fast progress in HIV-related basic research, antiretroviral therapy (ART) remains the most effective method to save AIDS patients' lives. Unfortunately, ART cannot be universally accessed, especially in developing countries, due to the lack of effective treatment monitoring diagnostics. Here, we present an inexpensive, rapid and portable micro-a-fluidic platform, which can streamline the process of an enzyme-linked immunosorbent assay (ELISA) in a fully automated manner for CD4 cell count. The micro-a-fluidic CD4 cell count is achieved by eliminating operational fluid flow via “moving the substrate”, as opposed to “flowing liquid” in traditional ELISA or microfluidic methods. This is the first demonstration of capturing and detecting cells from unprocessed whole blood using the enzyme-linked immunosorbent assay (ELISA) in a microfluidic channel. Combined with cell phone imaging, the presented micro-a-fluidic ELISA platform holds great promise for offering rapid CD4 cell count to scale up much needed ART in resource-constrained settings. The developed system can be extended to multiple areas for ELISA-related assays. PMID:24448112

  11. Micro-a-fluidics ELISA for rapid CD4 cell count at the point-of-care.

    PubMed

    Wang, ShuQi; Tasoglu, Savas; Chen, Paul Z; Chen, Michael; Akbas, Ragip; Wach, Sonya; Ozdemir, Cenk Ibrahim; Gurkan, Umut Atakan; Giguel, Francoise F; Kuritzkes, Daniel R; Demirci, Utkan

    2014-01-22

    HIV has become one of the most devastating pathogens in human history. Despite fast progress in HIV-related basic research, antiretroviral therapy (ART) remains the most effective method to save AIDS patients' lives. Unfortunately, ART cannot be universally accessed, especially in developing countries, due to the lack of effective treatment monitoring diagnostics. Here, we present an inexpensive, rapid and portable micro-a-fluidic platform, which can streamline the process of an enzyme-linked immunosorbent assay (ELISA) in a fully automated manner for CD4 cell count. The micro-a-fluidic CD4 cell count is achieved by eliminating operational fluid flow via "moving the substrate", as opposed to "flowing liquid" in traditional ELISA or microfluidic methods. This is the first demonstration of capturing and detecting cells from unprocessed whole blood using the enzyme-linked immunosorbent assay (ELISA) in a microfluidic channel. Combined with cell phone imaging, the presented micro-a-fluidic ELISA platform holds great promise for offering rapid CD4 cell count to scale up much needed ART in resource-constrained settings. The developed system can be extended to multiple areas for ELISA-related assays.

  12. The impact of diurnal variation on induced sputum cell counts in healthy adults

    PubMed Central

    2013-01-01

    Background Induced sputum cell counts are a non-invasive, reliable method for evaluating the presence, type, and degree of inflammation in the airways of the lungs. Current reference values for induced sputum cell counts in healthy adults do not account for the effects of circadian rhythm, including diurnal variation. The objective of this study was to describe the diurnal variation in induced sputum cell counts, compared between early morning and late afternoon, in healthy adult individuals. Methods 100 healthy adult subjects with no history of lung disease and normal bronchial reactivity proceeded with induced sputum testing at 7 am and 4 pm on different days. The order of testing was randomized. The cytotechnologist preparing and performing the cell counts was blinded to the sample collection time and subject characteristics. Results 65 subjects were included in the final analyses. There was no significant change in the total and differential sputum cell counts between the 7 am and 4 pm collections. There was good inter-observer agreement with respect to differential sputum cell count interpretation. Conclusions This is the largest study to assess the variation in induced sputum cell counts in healthy adult subjects at different times of the day. We found no significant change in total and differential sputum cell counts between the 7 am and 4 pm collection time points. This is in contrast to studies in asthmatics that demonstrated a circadian variation in sputum cell counts and other markers of inflammation, suggesting that fluctuations in airway inflammatory cells during the day are a disease-specific effect. PMID:23537329

  13. Accurate cell counts in live mouse embryos using optical quadrature and differential interference contrast microscopy

    NASA Astrophysics Data System (ADS)

    Warger, William C., II; Newmark, Judith A.; Zhao, Bing; Warner, Carol M.; DiMarzio, Charles A.

    2006-02-01

    Present imaging techniques used in in vitro fertilization (IVF) clinics are unable to produce accurate cell counts in developing embryos past the eight-cell stage. We have developed a method that has produced accurate cell counts in live mouse embryos ranging from 13-25 cells by combining Differential Interference Contrast (DIC) and Optical Quadrature Microscopy. Optical Quadrature Microscopy is an interferometric imaging modality that measures the amplitude and phase of the signal beam that travels through the embryo. The phase is transformed into an image of optical path length difference, which is used to determine the maximum optical path length deviation of a single cell. DIC microscopy gives distinct cell boundaries for cells within the focal plane when other cells do not lie in the path to the objective. Fitting an ellipse to the boundary of a single cell in the DIC image and combining it with the maximum optical path length deviation of a single cell creates an ellipsoidal model cell of optical path length deviation. Subtracting the model cell from the Optical Quadrature image will either show the optical path length deviation of the culture medium or reveal another cell underneath. Once all the boundaries are used in the DIC image, the subtracted Optical Quadrature image is analyzed to determine the cell boundaries of the remaining cells. The final cell count is produced when no more cells can be subtracted. We have produced exact cell counts on 5 samples, which have been validated by Epi-Fluorescence images of Hoechst stained nuclei.

  14. Variations in milk somatic cell count and haematologic values of dairy cows during lactation.

    PubMed

    Nikodémusz, E; Bedö, S; Pickler, A; Szép, P

    1994-01-01

    Variations in milk somatic cell count (SCC) and haematologic values were studied in a dairy cow colony of the Holstein-Friesian and Hungarian Red-Spotted breeds (n = 23) from May 1992 to July 1993. Milk and blood samples were taken approximately at monthly intervals and data were assigned into ten lunar months of lactation. After a maximum in month I, SCC dropped abruptly in month II and continued to decline through the subsequent four months, then it again tended to increase through months VII-X. The SCCs varied within the physiological range throughout the lactation period parallel with red blood cells and white blood cells constituting a major segment of the somatic cell population. Positive correlations were recorded between SCC and the blood variables (packed cell volume, red blood cell count, white blood cell count). The lactation pattern of SCC was comparable to previous observations.

  15. Preparing nuclei from cells in monolayer cultures suitable for counting and for following synchronized cells through the cell cycle.

    PubMed

    Butler, W B

    1984-08-15

    A procedure is described for preparing nuclei from cells in monolayer culture so that they may be counted using an electronic particle counter. It takes only 10 to 15 min, and consists of swelling the cells in hypotonic buffer and then lysing them with the quaternary ammonium salt, ethylhexadecyldimethylammonium bromide. The cells are completely lysed, yielding a suspension of clean single nuclei which is stable, free of debris, and easily counted. The method was developed for a cell line of epithelial origin (MCF-7), which is often difficult to trypsinize to single cells. It works equally well at all cell densities up to and beyond confluence, and has been used with a variety of cells in culture, including 3T3 cells, bovine macrophages, rat mammary epithelial cells, mouse mammary tumor cell lines, and human fibroblasts. The size of the nuclei produced by this procedure is related to their DNA content, and the method is thus suitable for following cultures of synchronized cells through the cell cycle, and for performing differential counts of cells with substantial differences in DNA content.

  16. Non-invasive, label-free cell counting and quantitative analysis of adherent cells using digital holography.

    PubMed

    Mölder, A; Sebesta, M; Gustafsson, M; Gisselson, L; Wingren, A Gjörloff; Alm, K

    2008-11-01

    Manual cell counting is time consuming and requires a high degree of skill on behalf of the person performing the count. Here we use a technique that utilizes digital holography, allowing label-free and completely non-invasive cell counting directly in cell culture vessels with adherent viable cells. The images produced can provide both quantitative and qualitative phase information from a single hologram. The recently constructed microscope Holomonitor (Phase Holographic Imaging AB, Lund, Sweden) combines the commonly used phase contrast microscope with digital holography, the latter giving us the possibility of achieving quantitative information on cellular shape, area, confluence and optical thickness. This project aimed at determining the accuracy and repeatability of cell counting measurements using digital holography compared to the conventional manual cell counting method using a haemocytometer. The collected data were also used to determine cell size and cellular optical thickness. The results show that digital holography can be used for non-invasive automatic cell counting as precisely as conventional manual cell counting.

  17. Machine Learning Based Single-Frame Super-Resolution Processing for Lensless Blood Cell Counting

    PubMed Central

    Huang, Xiwei; Jiang, Yu; Liu, Xu; Xu, Hang; Han, Zhi; Rong, Hailong; Yang, Haiping; Yan, Mei; Yu, Hao

    2016-01-01

    A lensless blood cell counting system integrating microfluidic channel and a complementary metal oxide semiconductor (CMOS) image sensor is a promising technique to miniaturize the conventional optical lens based imaging system for point-of-care testing (POCT). However, such a system has limited resolution, making it imperative to improve resolution from the system-level using super-resolution (SR) processing. Yet, how to improve resolution towards better cell detection and recognition with low cost of processing resources and without degrading system throughput is still a challenge. In this article, two machine learning based single-frame SR processing types are proposed and compared for lensless blood cell counting, namely the Extreme Learning Machine based SR (ELMSR) and Convolutional Neural Network based SR (CNNSR). Moreover, lensless blood cell counting prototypes using commercial CMOS image sensors and custom designed backside-illuminated CMOS image sensors are demonstrated with ELMSR and CNNSR. When one captured low-resolution lensless cell image is input, an improved high-resolution cell image will be output. The experimental results show that the cell resolution is improved by 4×, and CNNSR has 9.5% improvement over the ELMSR on resolution enhancing performance. The cell counting results also match well with a commercial flow cytometer. Such ELMSR and CNNSR therefore have the potential for efficient resolution improvement in lensless blood cell counting systems towards POCT applications. PMID:27827837

  18. Gene Signature of High White Blood Cell Count in B-Precursor Acute Lymphoblastic Leukemia.

    PubMed

    Edwards, Holly; Rubenstein, Mara; Dombkowski, Alan A; Caldwell, J Timothy; Chu, Roland; Xavier, Ana C; Thummel, Ryan; Neely, Melody; Matherly, Larry H; Ge, Yubin; Taub, Jeffrey W

    2016-01-01

    In this study we sought to identify genetic factors associated with the presenting white blood cell (WBC) count in B-precursor acute lymphoblastic leukemia (BP-ALL). Using ETV6-RUNX1-positive BP-ALL patient samples, a homogeneous subtype, we identified 16 differentially expressed genes based on the presenting WBC count (< 50,000/cumm vs > 50,000). We further confirmed that IL1R1, BCAR3, KCNH2, PIR, and ZDHHC23 were differentially expressed in a larger cohort of ETV6-RUNX1-negative BP-ALL patient samples. Statistical analysis demonstrated that expression levels of these genes could accurately categorize high and low WBC count subjects using two independent patient sets, representing positive and negative ETV6-RUNX1 cases. Further studies in leukemia cell line models will better delineate the role of these genes in regulating the white blood cell count and potentially identify new therapeutic targets.

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

    PubMed

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

    2002-02-05

    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 approximately 2 x 10(4) 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 indicates that there is a large number of cells, which then causes the aggregation stream to break up. Computer simulations indicated that stream breakup could be caused by CF decreasing cell-cell adhesion and/or increasing cell motility, and we observed that CF does indeed decrease cell-cell adhesion. We find here that CF increases cell motility. In Dictyostelium, motility is mediated by actin and myosin. CF increases the amounts of polymerized actin and the ABP-120 actin-crosslinking protein. Partially inhibiting motility by using drugs that interfere with actin polymerization reduces stream dissipation, resulting in fewer stream breaks and thus larger groups. CF also potentiates the phosphorylation and redistribution of myosin while repressing its basal level of assembly. The computer simulations indicated that a narrower distribution of group sizes results when a secreted factor modulates both adhesion and motility. CF thus seems to induce the morphogenesis of streams into evenly sized groups by increasing actin polymerization, ABP-120 levels, and myosin phosphorylation and decreasing adhesion and myosin polymerization.

  20. Lower white blood cell counts in elite athletes training for highly aerobic sports.

    PubMed

    Horn, P L; Pyne, D B; Hopkins, W G; Barnes, C J

    2010-11-01

    White cell counts at rest might be lower in athletes participating in selected endurance-type sports. Here, we analysed blood tests of elite athletes collected over a 10-year period. Reference ranges were established for 14 female and 14 male sports involving 3,679 samples from 937 females and 4,654 samples from 1,310 males. Total white blood cell counts and counts of neutrophils, lymphocytes and monocytes were quantified. Each sport was scaled (1-5) for its perceived metabolic stress (aerobic-anaerobic) and mechanical stress (concentric-eccentric) by 13 sports physiologists. Substantially lower total white cell and neutrophil counts were observed in aerobic sports of cycling and triathlon (~16% of test results below the normal reference range) compared with team or skill-based sports such as water polo, cricket and volleyball. Mechanical stress of sports had less effect on the distribution of cell counts. The lower white cell counts in athletes in aerobic sports probably represent an adaptive response, not underlying pathology.

  1. 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...

  2. 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 ...

  3. A microfluidic biochip for complete blood cell counts at the point-of-care

    PubMed Central

    Hassan, U.; Reddy, B.; Damhorst, G.; Sonoiki, O.; Ghonge, T.; Yang, C.; Bashir, R.

    2016-01-01

    Complete blood cell counts (CBCs) are one of the most commonly ordered and informative blood tests in hospitals. The results from a CBC, which typically include white blood cell (WBC) counts with differentials, red blood cell (RBC) counts, platelet counts and hemoglobin measurements, can have implications for the diagnosis and screening of hundreds of diseases and treatments. Bulky and expensive hematology analyzers are currently used as a gold standard for acquiring CBCs. For nearly all CBCs performed today, the patient must travel to either a hospital with a large laboratory or to a centralized lab testing facility. There is a tremendous need for an automated, portable point-of-care blood cell counter that could yield results in a matter of minutes from a drop of blood without any trained professionals to operate the instrument. We have developed microfluidic biochips capable of a partial CBC using only a drop of whole blood. Total leukocyte and their 3-part differential count are obtained from 10 μL of blood after on-chip lysing of the RBCs and counting of the leukocytes electrically using microfabricated platinum electrodes. For RBCs and platelets, 1 μL of whole blood is diluted with PBS on-chip and the cells are counted electrically. The total time for measurement is under 20 minutes. We demonstrate a high correlation of blood cell counts compared to results acquired with a commercial hematology analyzer. This technology could potentially have tremendous applications in hospitals at the bedside, private clinics, retail clinics and the developing world. PMID:26909365

  4. Mathematical modelling of adult hippocampal neurogenesis: effects of altered stem cell dynamics on cell counts and bromodeoxyuridine-labelled cells

    PubMed Central

    Ziebell, Frederik; Martin-Villalba, Ana; Marciniak-Czochra, Anna

    2014-01-01

    In the adult hippocampus, neurogenesis—the process of generating mature granule cells from adult neural stem cells—occurs throughout the entire lifetime. In order to investigate the involved regulatory mechanisms, knockout (KO) experiments, which modify the dynamic behaviour of this process, were conducted in the past. Evaluating these KOs is a non-trivial task owing to the complicated nature of the hippocampal neurogenic niche. In this study, we model neurogenesis as a multicompartmental system of ordinary differential equations based on experimental data. To analyse the results of KO experiments, we investigate how changes of cell properties, reflected by model parameters, influence the dynamics of cell counts and of the experimentally observed counts of cells labelled by the cell division marker bromodeoxyuridine (BrdU). We find that changing cell proliferation rates or the fraction of self-renewal, reflecting the balance between symmetric and asymmetric cell divisions, may result in multiple time phases in the response of the system, such as an initial increase in cell counts followed by a decrease. Furthermore, these phases may be qualitatively different in cells at different differentiation stages and even between mitotically labelled cells and all cells existing in the system. PMID:24598209

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

    PubMed Central

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

    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 indicates that there is a large number of cells, which then causes the aggregation stream to break up. Computer simulations indicated that stream breakup could be caused by CF decreasing cell–cell adhesion and/or increasing cell motility, and we observed that CF does indeed decrease cell–cell adhesion. We find here that CF increases cell motility. In Dictyostelium, motility is mediated by actin and myosin. CF increases the amounts of polymerized actin and the ABP-120 actin-crosslinking protein. Partially inhibiting motility by using drugs that interfere with actin polymerization reduces stream dissipation, resulting in fewer stream breaks and thus larger groups. CF also potentiates the phosphorylation and redistribution of myosin while repressing its basal level of assembly. The computer simulations indicated that a narrower distribution of group sizes results when a secreted factor modulates both adhesion and motility. CF thus seems to induce the morphogenesis of streams into evenly sized groups by increasing actin polymerization, ABP-120 levels, and myosin phosphorylation and decreasing adhesion and myosin polymerization. PMID:11818526

  6. 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.

  7. Two-dimensional versus three-dimensional cell counting: a practical perspective.

    PubMed

    Benes, F M; Lange, N

    2001-01-01

    In recent years, it has been argued by some neuroanatomists that three-dimensional (3-D) counting approaches must be used in studies of neural systems, so that 'unbiased' counts of neurons can be obtained. By contrast, two-dimensional (2-D) cell-counting methods are said to be 'assumption-based' and to yield inaccurate results. Working from the premise that all scientific methodologies are assumption-based and suffer from inherent biases, the current review considers the relative strengths and weaknesses of 2-D versus 3-D counting approaches. This comparison is from the standpoint of predictive performance with respect to bias, variance and fidelity to the actual spatial arrangements of cells in the tissue under study. When these considerations are taken, together with the human resources that are required in using either methodology, 2-D methods offer more practical alternatives that might even provide more scientifically accurate estimates compared with their 3-D counterparts.

  8. Method validation of circulating tumour cell enumeration at low cell counts

    PubMed Central

    2013-01-01

    Background Circulating tumour cells (CTC) are receiving increasing attention as prognostic, predictive and pharmacodynamic biomarkers in cancer patients. However, their clinical significance can be dependent on an accurate determination of CTC around cut-off values at low cell counts (<10 cells/7.5 ml). Consequently, we have conducted method validation of the CellSearch™ system focusing on clinical samples containing CTC in the cut-off region. Methods Analytical accuracy was first assessed employing quality controls (QC) and spiked healthy volunteer blood specimens. Results were analysed by β-expectation tolerance intervals (BETI). Inter-operator error (6 different readers) was then characterised in 38 different patient samples, 68% of which had ≤5 CTC and data were analysed by β-content γ-confidence tolerance intervals (BCTI). Results Results from QCs and spiked blood confirmed a 3-4-fold higher degree of imprecision at the low (48 cells, BETI = + 0.288/-0.345, β = 95%) compared to the high QC (987 cells, BETI = +0.065/-0.140, β = 95%). However, when data for individual analysts were interrogated characteristic systematic errors were detected. In the analysis of patient samples again individual analysts introduced a highly specific error into the interpretation of CTC images, which correlated to the level of training and experience. When readers were selected based on BETI and BCTI results, the high level of between-operator error (up to 170%) observed at CTC of ≤ 5 was reduced to < 30%. Conclusions Inter-operator variability in enumeration of CTC at low cell counts can be considerable, but is also potentially avoidable by following simple guidance steps. PMID:24024881

  9. Tomographic brain imaging with nucleolar detail and automatic cell counting

    NASA Astrophysics Data System (ADS)

    Hieber, Simone E.; Bikis, Christos; Khimchenko, Anna; Schweighauser, Gabriel; Hench, Jürgen; Chicherova, Natalia; Schulz, Georg; Müller, Bert

    2016-09-01

    Brain tissue evaluation is essential for gaining in-depth insight into its diseases and disorders. Imaging the human brain in three dimensions has always been a challenge on the cell level. In vivo methods lack spatial resolution, and optical microscopy has a limited penetration depth. Herein, we show that hard X-ray phase tomography can visualise a volume of up to 43 mm3 of human post mortem or biopsy brain samples, by demonstrating the method on the cerebellum. We automatically identified 5,000 Purkinje cells with an error of less than 5% at their layer and determined the local surface density to 165 cells per mm2 on average. Moreover, we highlight that three-dimensional data allows for the segmentation of sub-cellular structures, including dendritic tree and Purkinje cell nucleoli, without dedicated staining. The method suggests that automatic cell feature quantification of human tissues is feasible in phase tomograms obtained with isotropic resolution in a label-free manner.

  10. Tomographic brain imaging with nucleolar detail and automatic cell counting

    PubMed Central

    Hieber, Simone E.; Bikis, Christos; Khimchenko, Anna; Schweighauser, Gabriel; Hench, Jürgen; Chicherova, Natalia; Schulz, Georg; Müller, Bert

    2016-01-01

    Brain tissue evaluation is essential for gaining in-depth insight into its diseases and disorders. Imaging the human brain in three dimensions has always been a challenge on the cell level. In vivo methods lack spatial resolution, and optical microscopy has a limited penetration depth. Herein, we show that hard X-ray phase tomography can visualise a volume of up to 43 mm3 of human post mortem or biopsy brain samples, by demonstrating the method on the cerebellum. We automatically identified 5,000 Purkinje cells with an error of less than 5% at their layer and determined the local surface density to 165 cells per mm2 on average. Moreover, we highlight that three-dimensional data allows for the segmentation of sub-cellular structures, including dendritic tree and Purkinje cell nucleoli, without dedicated staining. The method suggests that automatic cell feature quantification of human tissues is feasible in phase tomograms obtained with isotropic resolution in a label-free manner. PMID:27581254

  11. Response of sheep lymphocytes to PHA: quantitation by nuclear volume measurement and cell counts (40764)

    SciTech Connect

    Chandra, P.; Chanana, A.D.; Joel, D.D.

    1980-03-01

    Phytohemagglutinin response of peripheral blood lymphocytes (PBL) of sheep was studied. Assessment of proliferative response was performed by determination of nuclear volumes and cell counts in cultures from 14 sheep and by incorporation of tritiated thymidine in cultures in four additional sheep. PBL of sheep were found to transform and proliferate with PHA similarly to human peripheral blood lymphocytes with minor differences. Quantitation of the proliferative response by determining the cell count and nuclear volumes provided more information on cell kinetics in culture than the commonly used isotope-labeled thymidine incorporation method.

  12. Total lymphocyte count is a reliable surrogate marker for CD4 cell counts after the first year of antiretroviral therapy: data from an Indonesian cohort study.

    PubMed

    de Jong, Marrigje A; Wisaksana, Rudi; Meijerink, Hinta; Indrati, Agnes; van de Ven, Andre J A M; Alisjahbana, Bachti; van Crevel, Reinout

    2012-05-01

    Many studies have evaluated the total lymphocyte count (TLC) as a cheap surrogate marker for CD4 cells in HIV-infected patients not receiving antiretroviral therapy (ART). We assessed whether TLC can replace CD4 cell counts in evaluating the immunological response to ART. In a cohort of patients in Indonesia TLC, if measured after at least 1-year ART, correctly identified patients with <200 CD4 cells, and reliably excluded immunological failure, obviating the need for CD4 cell measurement in 43% of patients.

  13. Negative prognostic impact of low absolute CD4(+) T cell counts in peripheral blood in mantle cell lymphoma.

    PubMed

    Zhang, Xin-Yu; Xu, Ji; Zhu, Hua-Yuan; Wang, Yan; Wang, Li; Fan, Lei; Wu, Yu-Jie; Li, Jian-Yong; Xu, Wei

    2016-10-01

    Tumor microenvironment and host immunity are closely related to outcome in patients with mantle cell lymphoma (MCL). However, few researchers have focused on the prognostic value of peripheral blood lymphocyte subsets counts. The purpose of this study was to investigate the prognostic value of lymphocyte subsets and absolute monocyte counts. Sixty-eight patients were analyzed retrospectively. Absolute CD4(+) T cell counts (ACD4C), CD8(+) T cell counts, nature killer cell counts, and CD4/CD8 ratios were assessed by peripheral blood flow cytometry and correlated with clinical parameters and long-term outcomes. The median follow-up for all patients was 21 months and the median survival time was 44 months. The overall survival (OS) rate at 1, 3, and 5 years was 80%, 51%, and 41%, respectively. In our cohort, high absolute monocyte count, and low ACD4C and CD4/CD8 ratio were associated with unfavorable OS (P = 0.029, P = 0.027, and P = 0.045, respectively) by univariate analysis. Multivariate analysis indicated that low ACD4C was a significant predictor of unfavorable OS (P = 0.004) independent of the simplified MCL International Prognostic Index (P = 0.048) in patients treated with or without rituximab (P = 0.011). Low CD4(+) T cell counts proved to be a significant predictor of unfavorable OS in patients with MCL.

  14. 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.

  15. A Novel Computerized Cell Count Algorithm for Biofilm Analysis

    PubMed Central

    Klinger-Strobel, Mareike; Suesse, Herbert; Fischer, Dagmar; Pletz, Mathias W.; Makarewicz, Oliwia

    2016-01-01

    Biofilms are the preferred sessile and matrix-embedded life form of most microorganisms on surfaces. In the medical field, biofilms are a frequent cause of treatment failure because they protect the bacteria from antibiotics and immune cells. Antibiotics are selected according to the minimal inhibitory concentration (MIC) based on the planktonic form of bacteria. Determination of the minimal biofilm eradicating concentration (MBEC), which can be up to 1,000-fold greater than the MIC, is not currently conducted as routine diagnostic testing, primarily because of the methodical hurdles of available biofilm assessing protocols that are time- and cost-consuming. Comparative analysis of biofilms is also limited as most quantitative methods such as crystal violet staining are indirect and highly imprecise. In this paper, we present a novel algorithm for assessing biofilm resistance to antibiotics that overcomes several of the limitations of alternative methods. This algorithm aims for a computer-based analysis of confocal microscope 3D images of biofilms after live/dead stains providing various biofilm parameters such as numbers of viable and dead cells and their vertical distributions within the biofilm, or biofilm thickness. The performance of this algorithm was evaluated using computer-simulated 2D and 3D images of coccal and rodent cells varying different parameters such as cell density, shading or cell size. Finally, genuine biofilms that were untreated or treated with nitroxoline or colistin were analyzed and the results were compared with quantitative microbiological standard methods. This novel algorithm allows a direct, fast and reproducible analysis of biofilms after live/dead staining. It performed well in biofilms of moderate cell densities in a 2D set-up however the 3D analysis remains still imperfect and difficult to evaluate. Nevertheless, this is a first try to develop an easy but conclusive tool that eventually might be implemented into routine

  16. 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...

  17. A semi-automated technique for labeling and counting of apoptosing retinal cells

    PubMed Central

    2014-01-01

    Background Retinal ganglion cell (RGC) loss is one of the earliest and most important cellular changes in glaucoma. The DARC (Detection of Apoptosing Retinal Cells) technology enables in vivo real-time non-invasive imaging of single apoptosing retinal cells in animal models of glaucoma and Alzheimer’s disease. To date, apoptosing RGCs imaged using DARC have been counted manually. This is time-consuming, labour-intensive, vulnerable to bias, and has considerable inter- and intra-operator variability. Results A semi-automated algorithm was developed which enabled automated identification of apoptosing RGCs labeled with fluorescent Annexin-5 on DARC images. Automated analysis included a pre-processing stage involving local-luminance and local-contrast “gain control”, a “blob analysis” step to differentiate between cells, vessels and noise, and a method to exclude non-cell structures using specific combined ‘size’ and ‘aspect’ ratio criteria. Apoptosing retinal cells were counted by 3 masked operators, generating ‘Gold-standard’ mean manual cell counts, and were also counted using the newly developed automated algorithm. Comparison between automated cell counts and the mean manual cell counts on 66 DARC images showed significant correlation between the two methods (Pearson’s correlation coefficient 0.978 (p < 0.001), R Squared = 0.956. The Intraclass correlation coefficient was 0.986 (95% CI 0.977-0.991, p < 0.001), and Cronbach’s alpha measure of consistency = 0.986, confirming excellent correlation and consistency. No significant difference (p = 0.922, 95% CI: −5.53 to 6.10) was detected between the cell counts of the two methods. Conclusions The novel automated algorithm enabled accurate quantification of apoptosing RGCs that is highly comparable to manual counting, and appears to minimise operator-bias, whilst being both fast and reproducible. This may prove to be a valuable method of quantifying apoptosing retinal

  18. Automated white blood cell counting via classification-free granulometric methods

    NASA Astrophysics Data System (ADS)

    Theera-Umpon, Nipon; Gader, Paul D.

    1999-03-01

    In this paper we describe an application of the granulometric mixing theorem to the problem of counting different types of white blood cells in bone marrow images. In principle, an iterative algorithm based on the mixing theorem can be used to count the proportion of cells in each class without explicitly segmenting and classifying them. The algorithm does not converge well for more than two classes. Therefore, a new algorithm based on the theorem is proposed. The proposed algorithm uses prior statistics to initially segment the mixed pattern spectrum and then applies the one-primitive mixing theorem to each initial component. Applying the mixing theorem to one class at a time results in better convergence. The counts produced by the proposed algorithm on 6 classes of cell -- Myeloblast, Promyelocyte, Myelocyte, Metamyelocyte, Band, and PMN -- are very close to the actual numbers; the deviation of the algorithm counts is not larger than deviation of counts produced by human experts. An important technical point is that, unlike previous algorithms, the proposed algorithm does not require prior knowledge of the total number of cells in an image.

  19. A comparison of 2 white blood cell count devices to aid judicious antibiotic prescribing.

    PubMed

    Casey, Janet R; Pichichero, Michael E

    2009-04-01

    A low or normal white blood cell (WBC) count is usually associated with viral illnesses. This study evaluated the reliability of a new point-of-care, inexpensive, WBC count device which requires only 10 microL (1 drop) of whole blood from a finger stick to an automated Cell-Dyn counter in a busy office practice setting and assessed its reliability to assist in avoiding antibiotic prescribing. A total of 120 acutely ill children and potential antibiotic recipients were studied from October 2007 to March 2008. The mean WBC count was 7.4x10(9)/L and 8.1x10( 9)/L for the new WBC device and the automated Cell-Dyn counter, respectively. The correlation between the 2 devices was high (r=.988, P=.005). A total of 88 children (73%) did not receive antibiotics and mean WBC was 7.2x10(9)/L. In all, 32 children (27%) received an antibiotic and 1 (3%) returned for a follow-up office visit for the same or a related illness. Of the 88 children with a low blood count who did not receive an antibiotic, 3 (3%) had return visit within 30 days and received an antibiotic. A simple and quick point-of-care WBC count device produces similar results as achievable with a Cell-Dyn counter for total WBCs and may assist in judicious antibiotic prescribing.

  20. Counting cells with a low-cost integrated microfluidics-waveguide sensor.

    PubMed

    Garcia, Daniel; Ghansah, Isaac; Leblanc, John; Butte, Manish J

    2012-03-01

    The capability to count cells from biofluids at low cost has important diagnostic implications in resource-poor settings. Many approaches have been developed to address this important need, and while most envision a low per-test cost, the detector instrument can be quite expensive. In this report, we present a novel device that enables low-cost and rapid counting of cells from a drop of blood. We demonstrate a shallow, buried, planar waveguide fabricated by ion exchange in glass that underlies a microfluidic structure for capturing cells. Laser light transmitted through the waveguide was attenuated by the number of metal nanoparticles tagged to the cells because of the interaction of the metal particles with the evanescent field of the waveguide. Calibration of the sensor using bead-tagged lymphocytes captured from human blood showed that the sensor could semi-quantitatively count as few as 100 cells/µL of blood. This technology enables the enumeration of specifically captured cells, allowing for a point-of-care, hand-held device for fast and affordable cell counting in screening, remote, or resource-poor settings.

  1. Concomitant spuriously elevated white blood cell count, a previously underestimated phenomenon in EDTA-dependent pseudothrombocytopenia.

    PubMed

    Xiao, Yufei; Xu, Yang

    2015-01-01

    The proportion and potential risk of concomitant spuriously elevated white blood cell count (SEWC) are underestimated in ethylenediaminetetraacetic acid (EDTA)-dependent pseudothrombocytopenia (PTCP). The proportion, kinetics and prevention of SEWC remain poorly understood. A total of 25 patients with EDTA-dependent PTCP were enrolled in this study. With the hematology analyzer Coulter LH 750, we determined the time courses of WBC count, WBC differential and platelet count in EDTA- and sodium citrate-anticoagulated blood, respectively. Blood smears were prepared to inspect the presence of platelet clumps using light microscopy. The effect of automatic instrumental correction on the extent of SEWC was evaluated. The proportion of SEWC was 92% in EDTA-dependent PTCP and 73.9% of SEWCs were within the normal range. The development of SEWC was time-dependent, and neutrophils and lymphocytes were the main subpopulations involved in SEWC. A strong and significant correlation (r = 0.9937, p < 0.001) was found between the increased WBC count and the decreased platelet count. Both corrected and uncorrected WBC counts at 15 minutes or later after blood collection in EDTA were significantly higher than their basal counts, respectively, p < 0.05. Interestingly, in citrated blood, WBC counts after blood collection were not significantly different from its basal counts, p > 0.05. A high proportion of concomitant SEWCs, which are mainly within normal range, are present in patients with EDTA-dependent PTCP. Proper interpretation of SEWC is crucial to avoid clinic errors. SEWC develops in a time-dependent pattern, although the Coulter LH 750 only partly mitigates the extent of SEWC, sodium citrate is able to effectively prevent SEWC.

  2. Factors influencing variation of bulk milk antibiotic residue occurrence, somatic cell count, and total bacterial count in dairy sheep flocks.

    PubMed

    Gonzalo, C; Carriedo, J A; García-Jimeno, M C; Pérez-Bilbao, M; de la Fuente, L F

    2010-04-01

    To study the variations of bulk tank milk variables in dairy ewe flocks and to identify the main target practices and flock groups to improve milk quality and safety, a total of 71,228 records of antibiotic residue (AR) and milk yield and 68,781 records of somatic cell count (SCC) and total bacterial count (TBC) were obtained over 5 yr from the same 209 dairy ewe flocks of the Assaf breed belonging to the Consortium for Ovine Promotion of Castilla-León (Spain). Based on a logistic regression model, year, month, semester, SCC, TBC, dry therapy, and milk yield significantly contributed to AR variation. High SCC was associated with increased AR violations. When antibiotic dry therapy was implemented, AR occurrence was higher than when this practice was not used. A polynomial monthly distribution throughout the year was observed for AR occurrence; the highest values were in autumn, coinciding with low milk yields per flock. Yearly occurrences drastically diminished from 2004 (1.36%) to 2008 (0.30%), probably as a result of effective educational programs. The mixed-model ANOVA of factors influencing variation in SCC and TBC indicated that year, month, AR, dry therapy group, milking type, and year interactions were significant variation factors for SCC and TBC; mathematical model accounted for 74.1 and 35.4% of total variance for each variable, respectively. Differences in management and hygiene practice caused significant SCC and TBC variations among flocks and within flocks throughout the 5-yr study. Over time, continuously dry treated flocks showed lower logSCC (5.80) and logTBC (4.92) than untreated (6.10 and 5.18, respectively) or discontinuously dry treated (6.01 and 5.05, respectively) flocks. Continuously dry treated flocks had lower AR occurrences than did discontinuously dry treated flocks. As a whole, AR occurrence and SCC and TBC bulk tank milk variables can be used for monitoring mammary health and milk hygiene and safety in dairy sheep throughout time.

  3. The Application of Bulk Tank Somatic Cell Counts to Monitoring Mastitis Levels in Dairy Herds

    PubMed Central

    Meek, A.H.; Barnum, D.A.

    1982-01-01

    The objective of this study was to investigate the feasibility of developing a system whereby measurements taken on bulk tank milk samples could be used to monitor the level of subclinical mastitis in dairy herds. The variables that were examined were the logarithmically transformed total somatic cell counts and percentages of cell volume in channel 8 (volumes from 89.2 to 178.3 µm3), the presence or absence of Streptococcus agalactiae and various husbandry/management factors including herdsize and the use of teat dips. Each of the use of actual monthly and rolling average bulk tank cell count determinations was investigated. It was found that the inclusion of all variables resulted in a correct classification of approximately 85% of herds and that no improvement was achieved by the use of rolling as opposed to actual monthly values. The inclusion of various husbandry/management practices improved the percentage correct classification to some extent over that achieved by the sole use of total somatic cell counts and percentages of cell volume in channel 8 when the herds were grouped on the basis of quarter infection rate (<10%, >10%) but not in the case of the cow infection rate categories (<20%, >20%). The use of both total cell counts and percentages of cell volume in channel 8 did not improve the overall predictive value over that achieved by the sole use of percentage of cell volume in channel 8 in the case of the quarter infection rate groupings but did to some extent in the case of the cow infection rate groupings. When the classification functions were applied prospectively and considering combinations of the two cell count determinations only, it was found that they were able to correctly classify, on the basis of the quarter infection rate groupings, approximately 75% of the study herds. It is concluded that the system described herein has limited application as a basis for selecting problem herds. PMID:7042053

  4. Electrical Counting and Sizing of Mammalian Cells in Suspension

    PubMed Central

    Gregg, E. C.; Steidley, K. David

    1965-01-01

    A recently developed method of determining the number and size of particles suspended in a conducting solution is to pump the suspension through a small orifice having an immersed electrode on each side to supply electrical current. The current changes due to the passage of particles of resistivity different from that of the solution. Theoretical expressions are developed which relate the current change caused by such particles to their volume and shape. It is found that most biological cells may be treated as dielectric particles whose capacitive effects are negligible. Electrolytic tank measurements on models confirm the theoretical development, and electric field plots of model orifices are used to predict the observed pulse shapes. An equivalent circuit of the orifice-electrode system is analyzed and shows that the current pulse may be made conductivity-independent when observed with a zero input impedance amplifier. PMID:5861698

  5. [Music therapy induced alternations in natural killer cell count and function].

    PubMed

    Hasegawa, Y; Kubota, N; Inagaki, T; Shinagawa, N

    2001-03-01

    The effects of music therapy on natural killer (NK) cell count and activity (NKCA) were studied in 19 persons. Alzheimer's disease, cerebrovessel disease and Parkinson's disease subjects were assigned to a music therapy. Blood samples were drawn at rest and after completion of music therapy. Music therapy did not change the number of circulating lymphocytes. The percentage of NK cells increased during music therapy, along with an increase in the NK cell activity. The proportion of T cells, CD4 and CD8 did not change significantly during music therapy. One hour after the music therapy session, plasma adrenaline increased but cortisol and noradrenalin did not change. The results indicate that music therapy can significantly increase NK cell count and activity. The change in NK cell and function were independent of neuro-degenerative diseases.

  6. RBC count

    MedlinePlus

    ... marrow disease that causes abnormal increase in RBCs ( polycythemia vera ) Your RBC count will increase for several ... Multiple myeloma Myelofibrosis Paroxysmal nocturnal hemoglobinuria (PNH) Peripheral Polycythemia vera Renal cell carcinoma Splenomegaly Transfusion reaction - hemolytic ...

  7. Reticulocyte count

    MedlinePlus

    Anemia - reticulocyte ... A higher than normal reticulocytes count may indicate: Anemia due to red blood cells being destroyed earlier than normal ( hemolytic anemia ) Bleeding Blood disorder in a fetus or newborn ( ...

  8. Chromosomal mosaicism in amniotic fluid cell cultures.

    PubMed Central

    Peakman, D C; Moreton, M F; Corn, B J; Robinson, A

    1979-01-01

    Over the past 6 years, using in situ processing methods, we have identified 32 cases of mosaicism in amniotic fluid cell cultures prepared from 1,100 samples. Two of these (45,X/46,XX and 46,XX/47,XX, + 21) were called true mosaics because multiple colonies demonstrated the same abnormal chromosome complement, and on subsequent evaluation of the newborn blood or fetal tissues, mosaicism was confirmed. Of the remaining cases, 29 were designated as pseudomosaics because only single or partial colonies exhibited an aberrant chromosome complement, 12 having a trisomy 2 line. In the final case, a double trisomy was demonstrated in only one of eight colonies in the first culture, but in the culture from a repeat sample an additional two colonies showed the same double trisomy. Since no abnormal cells were observed in infant blood, it was postulated that the mosaicism may only have been present in the extraembryonic tissues. It is our conviction that the use of these cloning methods should diminish the danger of misdiagnosis in genetic amniocentesis. PMID:453199

  9. AUTOMATED CELL COUNTING AND CLUSTER SEGMENTATION USING CONCAVITY DETECTION AND ELLIPSE FITTING TECHNIQUES

    PubMed Central

    Kothari, Sonal; Chaudry, Qaiser; Wang, May D

    2016-01-01

    This paper presents a novel, fast and semi-automatic method for accurate cell cluster segmentation and cell counting of digital tissue image samples. In pathological conditions, complex cell clusters are a prominent feature in tissue samples. Segmentation of these clusters is a major challenge for development of an accurate cell counting methodology. We address the issue of cluster segmentation by following a three step process. The first step involves pre-processing required to obtain the appropriate nuclei cluster boundary image from the RGB tissue samples. The second step involves concavity detection at the edge of a cluster to find the points of overlap between two nuclei. The third step involves segmentation at these concavities by using an ellipse-fitting technique. Once the clusters are segmented, individual nuclei are counted to give the cell count. The method was tested on four different types of cancerous tissue samples and shows promising results with a low percentage error, high true positive rate and low false discovery rate.

  10. Somatic cell counts, mastitis and milk production in selected Ontario dairy herds.

    PubMed Central

    Barnum, D A; Meek, A H

    1982-01-01

    Somatic cell counts were performed monthly on bulk tank milk samples for all producers in the Ontario counties of Hastings, Lennox/Addington and Prince Edward throughout 1978 and 1979. Other data were obtained via a structured questionnaire and from the records of the Ontario Milk Marketing Board. Many producers have not adopted practices that have been advocated for the integrated control of mastitis. For example, 43.3% of producers surveyed used single service paper towels, 63.3% regularly used teat dip and 56.5% dry cow therapy. The mean of the average monthly somatic cell count for all producers for 1978 was 621.1 x 10(3) cells/mL. This latter value was used to divide the producers into case (higher than average) and control (lower than average) groups. Control herds averaged 95.9 liters more shipped milk per cow per month than case herds. Milk from control herds averaged 0.22 percentage points higher than case herds for each of average fat and lactose, and 0.16 percentage points higher for protein. The linear regression of monthly shipped milk on the respective monthly bulk tank somatic cell count indicated a loss of 13.26 L/cow/month for each 100,000 increase in somatic cell count. PMID:7200385

  11. Differential white blood cell count and all-cause mortality in the Korean elderly.

    PubMed

    Kim, Kwang-Il; Lee, Jaebong; Heo, Nam Ju; Kim, Sejoong; Chin, Ho Jun; Na, Ki Young; Chae, Dong-Wan; Kim, Cheol-Ho; Kim, Suhnggwon

    2013-02-01

    The circulating white blood cell (WBC) count has been considered a good biomarker of systemic inflammation, but the predictive value of this inexpensive and universally obtained test result has not been fully explored in the elderly. The objective of this study was to assess the independent association of WBC count and its individual components with mortality in an elderly population. We studied a total of 9996 participants (age ≥65 years) who underwent routine health examinations at the 2 healthcare centers affiliated with Seoul National University. Mortality data were obtained from the National Statistics Office of Korea. The mean age of the study population was 69.7 (SD 4.3) years, and 5491 of the subjects (54.9%) were male. The median length of follow-up was 44.9 months (range, 1.2-78.7 months). There were 118 deaths (1.2%) during the follow-up period. The leading cause of death was cancer. Compared with the survivors, the deceased subjects were older, predominantly male, had increased levels of inflammatory markers, and had poor nutritional status. A significant difference in mortality was identified among patients in different WBC and WBC subtype quartile groups. Cox proportional hazards analysis indicated that monocyte count (HR: 5.18, 95% CI: 2.44-11.02) was a strongest predictor of all-cause mortality than total WBC count (HR: 1.57, 95% CI: 0.88-2.80), granulocyte count (HR: 2.11, 95% CI: 1.15-3.88), and lymphocyte count (HR: 1.11, 95% CI: 0.66-1.86), even after adjusting for possible confounding variables. Monocyte counts were associated with an increased risk of cardiovascular and cancer-related mortality in the elderly population. In conclusion, the total WBC count is an independent predictor of mortality in older adults, but the monocyte subtype provides greater predictive ability.

  12. Cell Counting in Human Endobronchial Biopsies - Disagreement of 2D versus 3D Morphometry

    PubMed Central

    Bratu, Vlad A.; Erpenbeck, Veit J.; Fehrenbach, Antonia; Rausch, Tanja; Rittinghausen, Susanne; Krug, Norbert; Hohlfeld, Jens M.; Fehrenbach, Heinz

    2014-01-01

    Question Inflammatory cell numbers are important endpoints in clinical studies relying on endobronchial biopsies. Assumption-based bidimensional (2D) counting methods are widely used, although theoretically design-based stereologic three-dimensional (3D) methods alone offer an unbiased quantitative tool. We assessed the method agreement between 2D and 3D counting designs in practice when applied to identical samples in parallel. Materials and Methods Biopsies from segmental bronchi were collected from healthy non-smokers (n = 7) and smokers (n = 7), embedded and sectioned exhaustively. Systematic uniform random samples were immunohistochemically stained for macrophages (CD68) and T-lymphocytes (CD3), respectively. In identical fields of view, cell numbers per volume unit (NV) were assessed using the physical disector (3D), and profiles per area unit (NA) were counted (2D). For CD68+ cells, profiles with and without nucleus were separately recorded. In order to enable a direct comparison of the two methods, the zero-dimensional CD68+/CD3+-ratio was calculated for each approach. Method agreement was tested by Bland-Altmann analysis. Results In both groups, mean CD68+/CD3+ ratios for NV and NA were significantly different (non-smokers: 0.39 and 0.68, p<0.05; smokers: 0.49 and 1.68, p<0.05). When counting only nucleated CD68+ profiles, mean ratios obtained by 2D and 3D counting were similar, but the regression-based Bland-Altmann analysis indicated a bias of the 2D ratios proportional to their magnitude. This magnitude dependent deviation differed between the two groups. Conclusions 2D counts of cell and nuclear profiles introduce a variable size-dependent bias throughout the measurement range. Because the deviation between the 3D and 2D data was different in the two groups, it precludes establishing a ‘universal conversion formula’. PMID:24663339

  13. Effect of leukaemic sera & cell-extracts on splenic colony counts (CFU-S).

    PubMed

    Gupta, S; Rusia, U; Agarwal, S; Sood, S K

    1991-08-01

    Sera and leukaemic cell extracts from patients of acute leukaemia were evaluated for their effect on the repopulating ability of the pluripotent stem cells and erythroid differentiation by an in vivo splenic colony count (CFU-S) technique. Normal donor marrow cells of mice were treated with sera and cell extracts from patients of acute leukaemic and healthy controls and injected in the recipient mice. The CFU-S performed on the seventh day to assess repopulating ability of the stem cell showed consistently lower CFU-S counts in the test groups, with leukaemic sera (P less than 0.01) as well as leukaemic cell-extracts (P less than 0.001). The erythroid differentiation assessed by 59Fe uptake by the spleens also showed significantly reduced counts in the two test groups (P less than 0.01 and less than 0.001 respectively). The results indicate that both leukaemic sera and cell-extracts exert a significant suppressive effect on the repopulating ability of the stem cells and on their erythroid differentiation.

  14. Fast automated yeast cell counting algorithm using bright-field and fluorescence microscopic images

    PubMed Central

    2013-01-01

    Background The faithful determination of the concentration and viability of yeast cells is important for biological research as well as industry. To this end, it is important to develop an automated cell counting algorithm that can provide not only fast but also accurate and precise measurement of yeast cells. Results With the proposed method, we measured the precision of yeast cell measurements by using 0%, 25%, 50%, 75% and 100% viability samples. As a result, the actual viability measured with the proposed yeast cell counting algorithm is significantly correlated to the theoretical viability (R2 = 0.9991). Furthermore, we evaluated the performance of our algorithm in various computing platforms. The results showed that the proposed algorithm could be feasible to use with low-end computing platforms without loss of its performance. Conclusions Our yeast cell counting algorithm can rapidly provide the total number and the viability of yeast cells with exceptional accuracy and precision. Therefore, we believe that our method can become beneficial for a wide variety of academic field and industries such as biotechnology, pharmaceutical and alcohol production. PMID:24215650

  15. Diagnostic value of circulating tumor cells in cerebrospinal fluid

    PubMed Central

    Ning, Mu; Chunhua, Ma; Yuan, Lv; Jinduo, Li; Bin, Wang; Liwei, Sun

    2016-01-01

    Abstract Objective To assess circulating tumor cells in cerebrospinal fluid as a diagnostic approach to identify meningeal metastasis in patients with non-small cell lung cancer by using tumor marker immunostaining–fluorescence in situ hybridization (TM-iFISH). Methods In 5 non-small cell lung cancer patients who were confirmed to have developed meningeal metastasis by cerebrospinal fluid cytology, 20 ml of cerebrospinal fluid was obtained through lumbar puncture, from which 7.5 ml was utilized for TM-iFISH to identify and quantitate circulating tumor cells, 10ml for cerebrospinal fluid cytology, and 2.5ml for detection of cerebrospinal fluid tumor markers. Results TM-iFISH examination identified 18 to 1,823 circulating tumor cells per 7.5ml cerebrospinal fluid. In contrast, cytology assessment revealed tumor cells in only 2 cases. The expression levels of cerebrospinal fluid tumor markers were all increased in all 5 patients when compared with their respective serum levels. Contrast-enhanced MRI scans demonstrated presence of meningeal metastasis in all 5 cases. Conclusion TM-iFISH may become a novel cerebrospinal fluid-based diagnostic strategy to identify circulating tumor cells and meningeal metastasis as compared to traditional diagnostic approaches, although its superior sensitivity and specificity needs to be confirmed through additional studies with a larger sample size.

  16. 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

  17. Total white blood cell count or neutrophil count predict ischemic stroke events among adult Taiwanese: report from a community-based cohort study

    PubMed Central

    2013-01-01

    Background Evidence about whether white blood cell (WBC) or its subtypes can act as a biomarker to predict the ischemic stroke events in the general population is scanty, particularly in Asian populations. The aim of this study is to establish the predictive ability of total WBC count or subtypes for long-term ischemic stroke events in the cohort population in Taiwan. Methods The Chin-Shan Community Cohort Study began from 1990 to 2007 by recruiting 1782 men and 1814 women of Chinese ethnicity. Following a total of 3416 participants free from ischemic stroke events at baseline for a median of 15.9 years; we documented 187 new incident cases. Results The multivariate relative risk for the comparison of the participants in the fifth and first WBC count quintiles was 1.67 (95% confidence interval [CI], 1.02–2.73; P for trend=0.03), and the corresponding relative risk for neutrophil count was 1.93 (95% CI, 1.13–3.29; P for trend=0.02). The discriminative ability by WBC and neutrophil counts were similar (area under the receiver operating characteristic curve, 0.600 for adding WBC, 0.610 for adding neutrophils, 0.595 for traditional risk factor model). In addition, the net reclassification improvement (NRI) values between the neutrophil and white blood cell count models were not significant (NRI, =-2.60%, P=0.35), indicating the similar discrimination performance for both WBC and neutrophil counts. Conclusions WBC and neutrophil count had a similar ability to predict the long-term ischemic stroke events among Taiwanese. PMID:23317415

  18. Longitudinal trends of total white blood cell and differential white blood cell counts of atomic bomb survivors.

    PubMed

    Hsu, Wan-Ling; Tatsukawa, Yoshimi; Neriishi, Kazuo; Yamada, Michiko; Cologne, John; Fujiwara, Saeko

    2010-01-01

    In studying the late health effects of atomic-bomb (A-bomb) survivors, earlier findings were that white blood cell (WBC) count increased with radiation dose in cross-sectional studies. However, a persistent effect of radiation on WBC count and other risk factors has yet to be confirmed. The objectives of the present study were 1) to examine the longitudinal relationship between A-bomb radiation dose and WBC and differential WBC counts among A-bomb survivors and 2) to investigate the potential confounding risk factors (such as age at exposure and smoking status) as well as modification of the radiation dose-response. A total of 7,562 A-bomb survivors in Hiroshima and Nagasaki were included in this study from 1964-2004. A linear mixed model was applied using the repeated WBC measurements. During the study period, a secular downward trend of WBC count was observed. Radiation exposure was a significant risk factor for elevated WBC and differential WBC counts over time. A significant increase of WBC counts among survivors with high radiation dose (> 2 Gy) was detected in men exposed below the age of 20 and in women regardless of age at exposure. Effects on WBC of low dose radiation remain unclear, however. Cigarette smoking produced the most pronounced effect on WBC counts and its impact was much larger than that of radiation exposure.

  19. Baseline CD4 Cell Counts of Newly Diagnosed HIV Cases in China: 2006–2012

    PubMed Central

    Tang, Houlin; Mao, Yurong; Shi, Cynthia X.; Han, Jing; Wang, Liyan; Xu, Juan; Qin, Qianqian; Detels, Roger; Wu, Zunyou

    2014-01-01

    Background Late diagnosis of HIV infection is common. We aim to assess the proportion of newly diagnosed HIV cases receiving timely baseline CD4 count testing and the associated factors in China. Methods Data were extracted from the Chinese HIV/AIDS Comprehensive Response Information Management System. Adult patients over 15 years old who had been newly diagnosed with HIV infection in China between 2006 and 2012 were identified. The study cohort comprised individuals who had a measured baseline CD4 count. Results Among 388,496 newly identified HIV cases, the median baseline CD4 count was 294 cells/µl (IQR: 130–454), and over half (N = 130,442, 58.8%) were less than 350 cells/µl. The median baseline CD4 count increased from 221 (IQR: 63–410) in 2006 to 314 (IQR: 159–460) in 2012. A slight majority of patients (N = 221,980, 57.1%) received baseline CD4 count testing within 6 months of diagnosis. The proportion of individuals who received timely baseline CD4 count testing increased significantly from 20.0% in 2006 to 76.9% in 2012. Factors associated with failing to receiving timely CD4 count testing were: being male (OR: 1.17, 95% CI: 1.15–1.19), age 55 years or older (OR:1.03, 95% CI: 1.00–1.06), educational attainment of primary school education or below (OR: 1.30, 95% CI: 1.28–1.32), infection with HIV through injection drug use (OR: 2.07, 95% CI: 2.02–2.12) or sexual contact and injection drug use (OR: 1.87, 95% CI: 1.76–1.99), diagnosis in a hospital (OR: 1.91, 95% CI: 1.88–1.95) or in a detention center (OR: 1.75, 95% CI: 1.70–1.80), and employment as a migrant worker (OR:1.55, 95% CI:1.53–1.58). Conclusion The proportion of newly identified HIV patients receiving timely baseline CD4 testing has increased significantly in China from 2006–2012. Continued effort is needed for further promotion of early HIV diagnosis and timely baseline CD4 cell count testing. PMID:24901790

  20. A method for high throughput determination of viable bacteria cell counts in 96-well plates

    PubMed Central

    2012-01-01

    Background There are several methods for quantitating bacterial cells, each with advantages and disadvantages. The most common method is bacterial plating, which has the advantage of allowing live cell assessment through colony forming unit (CFU) counts but is not well suited for high throughput screening (HTS). On the other hand, spectrophotometry is adaptable to HTS applications but does not differentiate between dead and living bacteria and has low sensitivity. Results Here, we report a bacterial cell counting method termed Start Growth Time (SGT) that allows rapid and serial quantification of the absolute or relative number of live cells in a bacterial culture in a high throughput manner. We combined the methodology of quantitative polymerase chain reaction (qPCR) calculations with a previously described qualitative method of bacterial growth determination to develop an improved quantitative method. We show that SGT detects only live bacteria and is sensitive enough to differentiate between 40 and 400 cells/mL. SGT is based on the re-growth time required by a growing cell culture to reach a threshold, and the notion that this time is proportional to the number of cells in the initial inoculum. We show several applications of SGT, including assessment of antibiotic effects on cell viability and determination of an antibiotic tolerant subpopulation fraction within a cell population. SGT results do not differ significantly from results obtained by CFU counts. Conclusion SGT is a relatively quick, highly sensitive, reproducible and non-laborious method that can be used in HTS settings to longitudinally assess live cells in bacterial cell cultures. PMID:23148795

  1. First steps to define murine amniotic fluid stem cell microenvironment

    PubMed Central

    Bertin, E.; Piccoli, M.; Franzin, C.; Spiro, G.; Donà, S.; Dedja, A.; Schiavi, F.; Taschin, E.; Bonaldo, P.; Braghetta, P.; De Coppi, P.; Pozzobon, M.

    2016-01-01

    Stem cell niche refers to the microenvironment where stem cells reside in living organisms. Several elements define the niche and regulate stem cell characteristics, such as stromal support cells, gap junctions, soluble factors, extracellular matrix proteins, blood vessels and neural inputs. In the last years, different studies demonstrated the presence of cKit+ cells in human and murine amniotic fluid, which have been defined as amniotic fluid stem (AFS) cells. Firstly, we characterized the murine cKit+ cells present both in the amniotic fluid and in the amnion. Secondly, to analyze the AFS cell microenvironment, we injected murine YFP+ embryonic stem cells (ESC) into the amniotic fluid of E13.5 wild type embryos. Four days after transplantation we found that YFP+ sorted cells maintained the expression of pluripotency markers and that ESC adherent to the amnion were more similar to original ESC in respect to those isolated from the amniotic fluid. Moreover, cytokines evaluation and oxygen concentration analysis revealed in this microenvironment the presence of factors that are considered key regulators in stem cell niches. This is the first indication that AFS cells reside in a microenvironment that possess specific characteristics able to maintain stemness of resident and exogenous stem cells. PMID:27845396

  2. Fluid flow plate for decreased density of fuel cell assembly

    DOEpatents

    Vitale, Nicholas G.

    1999-01-01

    A fluid flow plate includes first and second outward faces. Each of the outward faces has a flow channel thereon for carrying respective fluid. At least one of the fluids serves as reactant fluid for a fuel cell of a fuel cell assembly. One or more pockets are formed between the first and second outward faces for decreasing density of the fluid flow plate. A given flow channel can include one or more end sections and an intermediate section. An interposed member can be positioned between the outward faces at an interface between an intermediate section, of one of the outward faces, and an end section, of that outward face. The interposed member can serve to isolate the reactant fluid from the opposing outward face. The intermediate section(s) of flow channel(s) on an outward face are preferably formed as a folded expanse.

  3. Microfluidic cytometers with integrated on-chip optical systems for red blood cell and platelet counting.

    PubMed

    Zhao, Yingying; Li, Qin; Hu, Xiaoming; Lo, Yuhwa

    2016-11-01

    A microfluidic cytometer with integrated on-chip optical systems was designed for red blood cell (RBC) and platelet (PLT) counting. The design, fabrication, and characterization of the microfluidic cytometer with on-chip optical signal detection were described. With process using only a single mask, the device that integrates optical fibers and on-chip microlens with microfluidic channels on a polydimethylsiloxane layer by standard soft photolithography. This compact structure increased the sensitivity of the device and eliminated time-consuming free-space optical alignments. The microfluidic cytometer was used to count red blood cells and platelets. Forward scatter and extinction were collected simultaneously for each cell. Experimental results indicated that the microfluidic cytometer exhibited comparable performance with a conventional cytometer and demonstrated superior capacity to detect on-chip optical signals in a highly compact, simple, truly portable, and low-cost format that is well suitable for point-of-care clinical diagnostics.

  4. Joint Effect of Cigarette Smoking and Body Mass Index on White Blood Cell Count in Korean Adults

    PubMed Central

    Cho, A-Ra; Choi, Won-Jun; Kim, Shin-Hye; Shim, Jae-Yong

    2017-01-01

    Background White blood cell count is an independent risk factor for cardiovascular disease. Several lifestyle and metabolic factors such as cigarette smoking and obesity are known to be associated with an elevated white blood cell count. However, the joint effect of cigarette smoking and obesity on white blood cell count has not yet been fully described. Methods We explored the joint effect of cigarette smoking and obesity on white blood cell count using multiple logistic regression analyses after adjusting for confounding variables in a population-based, cross-sectional study of 416,065 Korean adults. Results Cigarette smoking and body mass index have a dose-response relationship with a higher white blood cell count, but no synergistic interaction is observed between them (men, P for interaction=0.797; women, P for interaction=0.311). Cigarette smoking and body mass index might have an additive combination effect on high white blood cell count. Obese male smokers were 2.36 times more likely and obese female smokers 2.35 times more likely to have a high white blood cell count when compared with normal body mass index non-smokers. Conclusion Cigarette smoking and body mass index are independently associated with an elevated white blood cell count in both men and women. PMID:28360982

  5. 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

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

    PubMed Central

    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

    2015-01-01

    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 (CD56bright/CD56dim) 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 CD56bright 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

  7. 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.

  8. Melanomas and Dysplastic Nevi Differ in Epidermal CD1c+ Dendritic Cell Count

    PubMed Central

    Dyduch, Grzegorz; Tyrak, Katarzyna Ewa; Glajcar, Anna; Szpor, Joanna

    2017-01-01

    Background. Dendritic cells could be involved in immune surveillance of highly immunogenic tumors such as melanoma. Their role in the progression melanocytic nevi to melanoma is however a matter of controversy. Methods. The number of dendritic cells within epidermis, in peritumoral zone, and within the lesion was counted on slides immunohistochemically stained for CD1a, CD1c, DC-LAMP, and DC-SIGN in 21 of dysplastic nevi, 27 in situ melanomas, and 21 invasive melanomas. Results. We found a significant difference in the density of intraepidermal CD1c+ cells between the examined lesions; the mean CD1c cell count was 7.00/mm2 for invasive melanomas, 2.94 for in situ melanomas, and 13.35 for dysplastic nevi. The differences between dysplastic nevi and melanoma in situ as well as between dysplastic nevi and invasive melanoma were significant. There was no correlation in number of positively stained cells between epidermis and dermis. We did not observe any intraepidermal DC-LAMP+ cells neither in melanoma in situ nor in invasive melanoma as well as any intraepidermal DC-SIGN+ cells in dysplastic nevi. Conclusion. It was shown that the number of dendritic cells differs between dysplastic nevi, in situ melanomas, and invasive melanomas. This could eventually suggest their participation in the development of melanoma. PMID:28331853

  9. Stem cells from amniotic fluid--Potential for regenerative medicine.

    PubMed

    Loukogeorgakis, Stavros P; De Coppi, Paolo

    2016-02-01

    Regenerative medicine has recently been established as an emerging field focussing on repair, replacement or regeneration of cells, tissues and whole organs. The significant recent advances in the field have intensified the search for novel sources of stem cells with potential for therapy. Recently, researchers have identified the amniotic fluid as an untapped source of stem cells that are multipotent, possess immunomodulatory properties and do not have the ethical and legal limitations of embryonic stem cells. Stem cells from the amniotic fluid have been shown to differentiate into cell lineages representing all three embryonic germ layers without generating tumours, which make them an ideal candidate for tissue engineering applications. In addition, their ability to engraft in injured organs and modulate immune and repair responses of host tissues suggest that transplantation of such cells may be useful for the treatment of various degenerative and inflammatory diseases affecting major tissues/organs. This review summarises the evidence on amniotic fluid cells over the past 15 years and explores the potential therapeutic applications of amniotic fluid stem cells and amniotic fluid mesenchymal stem cells.

  10. High-Throughput Method for Automated Colony and Cell Counting by Digital Image Analysis Based on Edge Detection

    PubMed Central

    Choudhry, Priya

    2016-01-01

    Counting cells and colonies is an integral part of high-throughput screens and quantitative cellular assays. Due to its subjective and time-intensive nature, manual counting has hindered the adoption of cellular assays such as tumor spheroid formation in high-throughput screens. The objective of this study was to develop an automated method for quick and reliable counting of cells and colonies from digital images. For this purpose, I developed an ImageJ macro Cell Colony Edge and a CellProfiler Pipeline Cell Colony Counting, and compared them to other open-source digital methods and manual counts. The ImageJ macro Cell Colony Edge is valuable in counting cells and colonies, and measuring their area, volume, morphology, and intensity. In this study, I demonstrate that Cell Colony Edge is superior to other open-source methods, in speed, accuracy and applicability to diverse cellular assays. It can fulfill the need to automate colony/cell counting in high-throughput screens, colony forming assays, and cellular assays. PMID:26848849

  11. High-Throughput Method for Automated Colony and Cell Counting by Digital Image Analysis Based on Edge Detection.

    PubMed

    Choudhry, Priya

    2016-01-01

    Counting cells and colonies is an integral part of high-throughput screens and quantitative cellular assays. Due to its subjective and time-intensive nature, manual counting has hindered the adoption of cellular assays such as tumor spheroid formation in high-throughput screens. The objective of this study was to develop an automated method for quick and reliable counting of cells and colonies from digital images. For this purpose, I developed an ImageJ macro Cell Colony Edge and a CellProfiler Pipeline Cell Colony Counting, and compared them to other open-source digital methods and manual counts. The ImageJ macro Cell Colony Edge is valuable in counting cells and colonies, and measuring their area, volume, morphology, and intensity. In this study, I demonstrate that Cell Colony Edge is superior to other open-source methods, in speed, accuracy and applicability to diverse cellular assays. It can fulfill the need to automate colony/cell counting in high-throughput screens, colony forming assays, and cellular assays.

  12. The effect of organic status and management practices on somatic cell counts on UK dairy farms.

    PubMed

    Haskell, M J; Langford, F M; Jack, M C; Sherwood, L; Lawrence, A B; Rutherford, K M D

    2009-08-01

    The numbers of organic dairy farms are increasing in the United Kingdom and in other parts of the world. On organic farms, the use of veterinary medicines is restricted. Because of this, there is concern that cow health is poorer on these farms. As udder health is primarily maintained by the use of antimicrobials, the effect of organic status on mastitis and somatic cell counts (SCC) is important to investigate. The aim of this study was therefore to determine whether the organic status and other management factors affect SCC. A group of 80 dairy farms was used in the study: 40 organic farms and 40 nonorganic farms. The farms were recruited in pairs, and each organic:nonorganic pair was matched for herd size, housing type, genetic merit for milk production and geographical location. Somatic cell count data were extracted from national databases for a standard year (2004), and analyzed using stepwise logistic regression models. The organic status of the farm did not appear in the final model, indicating no major influence of organic status on SCC. There were, however, several effects of management on SCC. Somatic cell counts were lower on farms where the udders were not cleaned or cleaned only when dirty. Somatic cell counts were also lower on farms that kept cows in larger management groups and where the majority, but not all cases of mastitis are treated with antimicrobials. It can be concluded that the control measures used on the organic farms in this study are at least as effective as those used on nonorganic farms in controlling SCC. Other management factors are influential and attention to these factors will allow farmers to reduce SCC.

  13. Metabolic activity of bacterial cell enumerated by direct viable count. [Escherichia coli; Salmonella enteritidis

    SciTech Connect

    Roszak, D.B.; Colwell, R.R.

    1987-12-01

    The direct viable count (DVC) method was modified by incorporation radiolabeled substrates in microautoradiographic analyses to assess bacterial survival in controlled laboratory microcosms. The DVC method, which permits enumeration of culturable and nonculturable cells, discriminates those cells that are responsive to added nutrients but in which division is inhibited by the addition of nalidixic acid. The resulting elongated cells represent all viable cells; this includes those that are culturable on routine media and those that are not. Escherichia coli and Salmonella enteritidis were employed in the microcosm studies, and radiolabeled substrates included (methyl-/sup 3/H) thymidine or (U-/sup 14/C) glutamic acid. Samples taken at selected intervals during the survival experiments were examined by epifluorescence microscopy to enumerate cells by the DVC and acridine orange direct count methods, as well as by culture methods. Good correlation was obtained for cell-associated metabolic activity, measured by microautoradiography and substrate responsiveness (by the DVC method) at various stages of survival. Of the cells responsive to nutrients by the DVC method, ca. 90% were metabolically active by the microautoradiographic method. No significant difference was observed between DVC enumerations with or without added radiolabeled substrate.

  14. Prognostic significance of peripheral monocyte count in patients with extranodal natural killer/T-cell lymphoma

    PubMed Central

    2013-01-01

    Background Extranodal natural killer/T-cell lymphoma (ENKL) has heterogeneous clinical manifestations and prognosis. This study aims to evaluate the prognostic impact of absolute monocyte count (AMC) in ENKL, and provide some immunologically relevant information for better risk stratification in patients with ENKL. Methods Retrospective data from 163 patients newly diagnosed with ENKL were analyzed. The absolute monocyte count (AMC) at diagnosis was analyzed as continuous and dichotomized variables. Independent prognostic factors of survival were determined by Cox regression analysis. Results The AMC at diagnosis were related to overall survival (OS) and progression-free survival (PFS) in patients with ENKL. Multivariate analysis identified AMC as independent prognostic factors of survival, independent of International Prognostic Index (IPI) and Korean prognostic index (KPI). The prognostic index incorporating AMC and absolute lymphocyte count (ALC), another surrogate factor of immune status, could be used to stratify all 163 patients with ENKL into different prognostic groups. For patients who received chemotherapy followed by radiotherapy (102 cases), the three AMC/ALC index categories identified patients with significantly different survivals. When superimposed on IPI or KPI categories, the AMC/ALC index was better able to identify high-risk patients in the low-risk IPI or KPI category. Conclusion The baseline peripheral monocyte count is shown to be an effective prognostic indicator of survival in ENKL patients. The prognostic index related to tumor microenvironment might be helpful to identify high-risk patients with ENKL. PMID:23638998

  15. Associations between White Blood Cell Count and the Development of Incidental Nonalcoholic Fatty Liver Disease

    PubMed Central

    Chung, Goh Eun; Kim, Donghee; Kwak, Min-Sun; Yang, Jong In; Chung, Su Jin; Yang, Sun Young; Kim, Joo Sung

    2016-01-01

    Aims. Chronic low-grade inflammation is thought to be associated with the pathogenesis of nonalcoholic fatty liver disease (NAFLD). This study aimed to determine the association between serum white blood cell (WBC) counts and the development of incidental NAFLD. Methods. In this retrospective longitudinal cohort study, we recruited participants who underwent abdominal ultrasonography and blood samplings during medical checkups in both 2005 and 2010. A total of 2,216 subjects were included in our analyses. Results. The prevalence of NAFLD in 2010 increased steadily in conjunction with increasing WBC counts in 2005 after adjustment for body mass index (BMI) [odds ratio (OR) 2.44, 95% confidence interval (CI) = 1.49–4.00 for women and OR 2.42, 95% CI = 1.61–3.63 for men, lowest quartile versus highest quartile]. Multivariate regression analysis after adjusting for age, BMI, hypertension, smoking, triglycerides, HDL cholesterol, and glucose levels revealed that NAFLD was significantly associated with the highest WBC quartile compared to the lowest quartile [OR 1.85, 95% CI, 1.10−3.10 for women and OR 1.68, 95% CI, 1.08−2.61 for men]. Conclusions. We demonstrated that the risk of developing NAFLD was significantly associated with WBC counts independently of metabolic factors. This finding provides novel evidence indicating that serum WBC counts may be potential surrogate markers of NAFLD. PMID:28070183

  16. Associations between White Blood Cell Count and the Development of Incidental Nonalcoholic Fatty Liver Disease.

    PubMed

    Chung, Goh Eun; Yim, Jeong Yoon; Kim, Donghee; Kwak, Min-Sun; Yang, Jong In; Chung, Su Jin; Yang, Sun Young; Kim, Joo Sung

    2016-01-01

    Aims. Chronic low-grade inflammation is thought to be associated with the pathogenesis of nonalcoholic fatty liver disease (NAFLD). This study aimed to determine the association between serum white blood cell (WBC) counts and the development of incidental NAFLD. Methods. In this retrospective longitudinal cohort study, we recruited participants who underwent abdominal ultrasonography and blood samplings during medical checkups in both 2005 and 2010. A total of 2,216 subjects were included in our analyses. Results. The prevalence of NAFLD in 2010 increased steadily in conjunction with increasing WBC counts in 2005 after adjustment for body mass index (BMI) [odds ratio (OR) 2.44, 95% confidence interval (CI) = 1.49-4.00 for women and OR 2.42, 95% CI = 1.61-3.63 for men, lowest quartile versus highest quartile]. Multivariate regression analysis after adjusting for age, BMI, hypertension, smoking, triglycerides, HDL cholesterol, and glucose levels revealed that NAFLD was significantly associated with the highest WBC quartile compared to the lowest quartile [OR 1.85, 95% CI, 1.10-3.10 for women and OR 1.68, 95% CI, 1.08-2.61 for men]. Conclusions. We demonstrated that the risk of developing NAFLD was significantly associated with WBC counts independently of metabolic factors. This finding provides novel evidence indicating that serum WBC counts may be potential surrogate markers of NAFLD.

  17. Experience with local lymph node assay performance standards using standard radioactivity and nonradioactive cell count measurements.

    PubMed

    Basketter, David; Kolle, Susanne N; Schrage, Arnhild; Honarvar, Naveed; Gamer, Armin O; van Ravenzwaay, Bennard; Landsiedel, Robert

    2012-08-01

    The local lymph node assay (LLNA) is the preferred test for identification of skin-sensitizing substances by measuring radioactive thymidine incorporation into the lymph node. To facilitate acceptance of nonradioactive variants, validation authorities have published harmonized minimum performance standards (PS) that the alternative endpoint assay must meet. In the present work, these standards were applied to a variant of the LLNA based on lymph node cell counts (LNCC) run in parallel as a control with the standard LLNA with radioactivity measurements, with threshold concentrations (EC3) being determined for the sensitizers. Of the 22 PS chemicals tested in this study, 21 yielded the same results from standard radioactivity and cell count measurements; only 2-mercaptobenzothiazole was positive by LLNA but negative by LNCC. Of the 16 PS positives, 15 were positive by LLNA and 14 by LNCC; methylmethacrylate was not identified as sensitizer by either of the measurements. Two of the six PS negatives tested negative in our study by both LLNA and LNCC. Of the four PS negatives which were positive in our study, chlorobenzene and methyl salicylate were tested at higher concentrations than the published PS, whereas the corresponding concentrations resulted in consistent negative results. Methylmethacrylate and nickel chloride tested positive within the concentration range used for the published PS. The results indicate cell counts and radioactive measurements are in good accordance within the same LLNA using the 22 PS test substances. Comparisons with the published PS results may, however, require balanced analysis rather than a simple checklist approach.

  18. Do white-cell count spikes predict agranulocytosis in clozapine recipients?

    PubMed

    Alvir, J M; Lieberman, J A; Safferman, A Z

    1995-01-01

    The use of clozapine is limited by the risk of agranulocytosis. The incidence of agranulocytosis after 1 year was .80 percent in 11,555 patients registered in the Clozaril Patient Management System (CPMS) who received clozapine from February 1990 to April 1991. We noticed a tendency for white-cell counts to spike upward before agranulocytosis occurred. We analyzed the CPMS data to test whether a white-cell count spike at least 15 percent above the previous measurement predicted agranulocytosis within 75 days, with a 21-day lag to allow white-cell counts to decline to levels indicative of agranulocytosis. The occurrence of a spike, entered as a time-dependent covariate in proportional hazards regression, significantly predicted development of agranulocytosis (risk ratio, 3.02; 95% confidence interval, 1.38 to 6.57). Spikes were sensitive though nonspecific predictors, occurring frequently in patients who did not develop agranulocytosis. These results, while exploratory, indicate the potential usefulness of these spikes as guidelines to govern the use of clozapine.

  19. Blood cell counting and classification by nonflowing laser light scattering method

    NASA Astrophysics Data System (ADS)

    Yang, Ye; Zhang, Zhenxi; Yang, Xinhui; Jiang, Dazong; Yeo, Joon Hock

    1999-11-01

    A new non-flowing laser light scattering method for counting and classifying blood cells is presented. A linear charge- coupled device with 1024 elements is used to detect the scattered light intensity distribution of the blood cells. A pinhole plate is combined with the CCD to compete the focusing of the measurement system. An isotropic sphere is used to simulate the blood cell. Mie theory is used to describe the scattering of blood cells. In order to inverse the size distribution of blood cells from their scattered light intensity distribution, Powell method combined with precision punishment method is used as a dependent model method for measurement red blood cells and blood plates. Non-negative constraint least square method combined with Powell method and precision punishment method is used as an independent model for measuring white blood cells. The size distributions of white blood cells and red blood cells, and the mean diameter of red blood cells are measured by this method. White blood cells can be divided into three classes: lymphocytes, middle-sized cells and neutrocytes according to their sizes. And the number of blood cells in unit volume can also be measured by the linear dependence of blood cells concentration on scattered light intensity.

  20. Blood cell counting and classification by nonflowing laser light scattering method.

    PubMed

    Yang, Ye; Zhang, Zhenxi; Yang, Xinhui; Yeo, Joon Hock; Jiang, LiJun; Jiang, Dazong

    2004-01-01

    We present a nonflowing laser light scattering method for automatically counting and classifying blood cells. A linear charge-coupled device (CCD) and a silicon photoelectric cell (which is placed behind a pinhole plate on the CCD) form a double-detector structure: the CCD is used to detect the scattered light intensity distribution of the blood cells and the silicon photoelectric cell to complete the focusing process. An isotropic sphere, with relative refractivity near 1, is used to model the blood cell. Mie theory is used to describe the scattering of white blood cells and platelets, and anomalous diffraction, red blood cells. To obtain the size distribution of blood cells from their scattered light intensity distribution, the nonnegative constraint least-squares (NNLS) method combined with the Powell method and the precision punishment method are used. Both numerical simulation and experimental results are presented. This method can be used not only to measure the mean and the distribution of red blood cell size, but also to divide the white blood cells into three classes: lymphocytes, middle-sized cells, and neutrocytes. The experimental results show a linear relationship between the blood cell (both white and red blood cells) concentration and the scattered light intensity, and therefore, the number of blood cells in a unit volume can be determined from this relationship.

  1. Association Between White Blood Cell Count Following Radiation Therapy With Radiation Pneumonitis in Non-Small Cell Lung Cancer

    SciTech Connect

    Tang, Chad; Gomez, Daniel R.; Wang, Hongmei; Levy, Lawrence B.; Zhuang, Yan; Xu, Ting; Nguyen, Quynh; Komaki, Ritsuko; Liao, Zhongxing

    2014-02-01

    Purpose: Radiation pneumonitis (RP) is an inflammatory response to radiation therapy (RT). We assessed the association between RP and white blood cell (WBC) count, an established metric of systemic inflammation, after RT for non-small cell lung cancer. Methods and Materials: We retrospectively analyzed 366 patients with non-small cell lung cancer who received ≥60 Gy as definitive therapy. The primary endpoint was whether WBC count after RT (defined as 2 weeks through 3 months after RT completion) was associated with grade ≥3 or grade ≥2 RP. Median lung volume receiving ≥20 Gy (V{sub 20}) was 31%, and post-RT WBC counts ranged from 1.7 to 21.2 × 10{sup 3} WBCs/μL. Odds ratios (ORs) associating clinical variables and post-RT WBC counts with RP were calculated via logistic regression. A recursive-partitioning algorithm was used to define optimal post-RT WBC count cut points. Results: Post-RT WBC counts were significantly higher in patients with grade ≥3 RP than without (P<.05). Optimal cut points for post-RT WBC count were found to be 7.4 and 8.0 × 10{sup 3}/μL for grade ≥3 and ≥2 RP, respectively. Univariate analysis revealed significant associations between post-RT WBC count and grade ≥3 (n=46, OR=2.6, 95% confidence interval [CI] 1.4‒4.9, P=.003) and grade ≥2 RP (n=164, OR=2.0, 95% CI 1.2‒3.4, P=.01). This association held in a stepwise multivariate regression. Of note, V{sub 20} was found to be significantly associated with grade ≥2 RP (OR=2.2, 95% CI 1.2‒3.4, P=.01) and trended toward significance for grade ≥3 RP (OR=1.9, 95% CI 1.0-3.5, P=.06). Conclusions: Post-RT WBC counts were significantly and independently associated with RP and have potential utility as a diagnostic or predictive marker for this toxicity.

  2. Composition, indigenous proteolytic enzymes and coagulating behaviour of ewe milk as affected by somatic cell count.

    PubMed

    Albenzio, Marzia; Santillo, Antonella; Caroprese, Mariangela; Schena, Laura; Russo, Donatella Esterina; Sevi, Agostino

    2011-11-01

    This study was undertaken to assess the effect of somatic cell count in ewe milk on i) composition and hygienic traits; ii) plasmin, cathepsin and elastase activities; iii) leukocyte differential count; iv) renneting parameters. Individual ewe milk samples were grouped according to somatic cell count (SCC) into five classes: SC300 (<300 000 cells/ml), SC500 (from 301 000 to 500 000 cells/ml), SC1000 (from 501 000 to 1 000 000 cells/ml), SC2000 (from 1 001 000 to 2 000 000 cells/ml) and SC>2000 (>2 001 000 cells/ml). Individual milk samples were analysed for pH, chemical composition, microbial features, indigenous proteolytic enzymes, differential leukocyte population, and renneting parameters. Milk yield, lactose, protein, non casein nitrogen, microbial features were affected by SCC level. Plasmin and elastase activities were the highest in samples with more than 1 000 000 cells/ml; plasmin had intermediate values in samples with 300 000 to 1 000 000 cells/ml and the lowest in samples with less than 300 000 cells/ml of milk. Cathepsin D showed significantly lower values in SC300 and SC1000 classes than in SC500, SC2000 and SC>2000 classes. The highest percentages of lymphocyte were found in samples with less than 1 000 000 cells/ml, while the highest levels of polymorphonuclear leukocyte were found in samples with more than 1 000 000 cells/ml of milk. Longer clotting time was found in SC>2000 samples, while reduced clot firmness was observed in SC500 and SC>2000 samples. Results on milk yield and on compositional parameters evidenced an impairment of udder efficiency in ewe milk samples starting from 300 000 cells/ml. Plasmin activity in milk can be considered as a marker of the synthetic and secreting ability of the mammary gland; furthermore plasmin and elastase were consistent with the health status of the udder. Finally cathepsin D played a role in the worsening of renneting properties of ewe milk.

  3. Peripheral Immune Cell Counts and Advanced Imaging as Biomarkers of Stroke Outcome

    PubMed Central

    Pagram, Heather; Bivard, Andrew; Lincz, Lisa F.; Levi, Christopher

    2016-01-01

    Introduction Circulating neutrophil and lymphocyte levels may be modifiable outcome predictors of ischemic stroke. We sought to compare these immune cell parameters with advanced imaging assessment and the 90-day clinical outcome. Methods We used a retrospectively collected cohort of consecutive ischemic stroke patients presenting within 4.5 h of symptom onset who had acute CT perfusion and routine blood collection before treatment with intravenous thrombolysis and 24-hour MRI scanning at the John Hunter Hospital. Full blood counts were performed acutely at 24 h and 7 days. Patient outcomes were assed at 90 days after stroke with the modified Rankin Scale (mRS). Results Overall, 142 patients were assessed during the study period. Patients with a poor outcome (mRS 3-6) had increased neutrophils (44% increase, p = 0.016), decreased lymphocytes (7% decrease, p = 0.491) and an increased lymphocyte-to-neutrophil ratio (196% increase, p < 0.001). Patients with good outcomes (mRS 0-2) did not have significant changes in their full blood counts. There was no relationship between the neutrophil count at 24 h and penumbral volume (r2 = 0.217, p = 0.212), reperfusion (r2 = 0.111, p = 0.085), or core growth (r2 = 0.297, p = 0.107). A backward multivariate analysis containing the 24-hour core volume and 24-hour neutrophil count was strongly related to the 3-month outcome (r2 = 0.477, area under the curve = 0.902, p < 0.001). Conclusions Peripheral neutrophils have potential as a biomarker of outcome when used in conjunction with advanced imaging. Peripherally measured neutrophil counts change significantly over time after stroke and may be potential targets for immunomodulatory therapy in patients with a severe stroke or a large infarct volume. PMID:27771707

  4. Bacteriology and somatic cell counts in milk samples from ewes on a Scottish farm.

    PubMed

    Hariharan, Harry; Donachie, Willie; Macaldowie, Colin; Keefe, Greg

    2004-07-01

    Milk samples from 50 sheep on a single Scottish research farm were collected weekly for 10 wk postpartum. Samples were analyzed for somatic cell counts (SCC) each week and bacteriologic culture was done for 7 of the 10 wk. A total of 492 udder half samples were cultured, of which 467 had corresponding cell count data. Statistical analysis on complete SCC and culture data showed no association between SCC and bacterial isolation, even when more than 10 colonies of a single bacterial species were present. Only 3.6% of the samples were simultaneously positive for high count (> 10 colonies from 0.01 mL of milk) of any one bacterial species and high SCC (> 1 x 10(6)/mL). The bacteria recovered were: Staphylococcus equorum (19 times), S. xylosus (7 times), S. simulans (6 times), Streptococcus uberis (3 times) and other streptococci (4 times), Mannheimia (Pasteurella) haemolytica (2 times), Staphylococcus aureus (1 time), S. capitis (1 time), and Enterococcus faecium (1 time). There was an association between the test day and SCC, with higher SCC values in the first 2 wk. In addition, significantly higher SCC values were found in the oldest animals compared to the other age groups.

  5. An integrated microfluidic platform for rapid tumor cell isolation, counting and molecular diagnosis.

    PubMed

    Hung, Lien-Yu; Chuang, Ying-Hsin; Kuo, Hsin-Tzu; Wang, Chih-Hung; Hsu, Keng-Fu; Chou, Cheng-Yang; Lee, Gwo-Bin

    2013-04-01

    Ovarian cancer is the second most common of the gynecological cancers in Taiwan. It is challenging to diagnose at an early stage when proper treatment is the most effective. It is well recognized that the detection of tumor cells (TCs) is critical for determining cancer growth stages and may provide important information for accurate diagnosis and even prognosis. In this study, a new microfluidic platform integrated with a moving-wall micro-incubator, a micro flow cytometer and a molecular diagnosis module performed automated identification of ovarian cancer cells. By efficiently mixing the cells and immunomagnetic beads coated with specific antibodies, the target TCs were successfully isolated from the clinical samples. Then counting of the target cells was achieved by a combination of the micro flow cytometer and an optical detection module and showed a counting accuracy as high as 92.5 %. Finally, cancer-associated genes were amplified and detected by the downstream molecular diagnosis module. The fluorescence intensity of specific genes (CD24 and HE4) associated with ovarian cancer was amplified by the molecular diagnosis module and the results were comparable to traditional slab-gel electrophoresis analysis, with a limit of detection around 10 TCs. This integrated microfluidic platform realized the concept of a "lab-on-a-chip" and had advantages which included automation, disposability, lower cost and rapid diagnosis and, therefore, may provide a promising approach for the fast and accurate detection of cancer cells.

  6. Effects of Fluid Shear Stress on Cancer Stem Cell Viability

    NASA Astrophysics Data System (ADS)

    Sunday, Brittney; Triantafillu, Ursula; Domier, Ria; Kim, Yonghyun

    2014-11-01

    Cancer stem cells (CSCs), which are believed to be the source of tumor formation, are exposed to fluid shear stress as a result of blood flow within the blood vessels. It was theorized that CSCs would be less susceptible to cell death than non-CSCs after both types of cell were exposed to a fluid shear stress, and that higher levels of fluid shear stress would result in lower levels of cell viability for both cell types. To test this hypothesis, U87 glioblastoma cells were cultured adherently (containing smaller populations of CSCs) and spherically (containing larger populations of CSCs). They were exposed to fluid shear stress in a simulated blood flow through a 125-micrometer diameter polyetheretherketone (PEEK) tubing using a syringe pump. After exposure, cell viability data was collected using a BioRad TC20 Automated Cell Counter. Each cell type was tested at three physiological shear stress values: 5, 20, and 60 dynes per centimeter squared. In general, it was found that the CSC-enriched U87 sphere cells had higher cell viability than the CSC-depleted U87 adherent cancer cells. Interestingly, it was also observed that the cell viability was not negatively affected by the higher fluid shear stress values in the tested range. In future follow-up studies, higher shear stresses will be tested. Furthermore, CSCs from different tumor origins (e.g. breast tumor, prostate tumor) will be tested to determine cell-specific shear sensitivity. National Science Foundation Grant #1358991 supported the first author as an REU student.

  7. 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

  8. Differences in lymphocyte subpopulations and cell counts before and after experimentally induced swine dysentery.

    PubMed

    Jonasson, Robert; Johannisson, Anders; Jacobson, Magdalena; Fellström, Claes; Jensen-Waern, Marianne

    2004-04-01

    The aim of this study was to examine the levels of circulating leukocytes and lymphocyte subpopulations before and immediately after experimentally induced swine dysentery. Twenty-one healthy crossbred pigs (approximately 22 kg) were orally inoculated with Brachyspira hyodysenteriae. Blood was sampled before inoculation and when clinical signs of swine dysentery occurred. Pigs that remained healthy were sampled when killed. Total and differential white blood cell counts were performed, and lymphocyte subpopulations were analysed using flow cytometry. Following a mean incubation period of 13 days, 12 pigs developed swine dysentery, whereas nine remained healthy throughout the study. Before inoculation, pigs that subsequently developed swine dysentery displayed higher levels of circulating gamma delta T cells (mean +/- se; 30.7 +/- 3.5 %) compared with pigs that remained healthy (14.9 +/- 1.4 %). Sick animals also displayed lower levels of CD8 cells (24.6 +/- 1.5 %), cytotoxic/suppressor T cells (10.9 +/- 1.3 %) and CD4 CD8 T cells (8.1 +/- 1.0 %) than the pigs that remained healthy (34.9 +/- 3.1 %; 17.6 +/- 2.0 %; 13.6 +/- 2.3 %). No difference was observed in leukocyte counts before inoculation. At onset of swine dysentery, there was an increase in monocytes (from 1.5 +/- 0.2 x 10 to 3.8 +/- 0.5 x 10 l) and CD4 CD8 T cells (from 5.8 +/- 0.9 to 8.9 +/- 0.7 %). In conclusion, gamma delta T cells and CD8 cells may be associated with susceptibility to experimentally induced swine dysentery, whereas monocytes and CD4 CD8 T cells appear to be the major responding leukocytes during the disease.

  9. Evaluation of nano-magnetic fluid on malignant glioma cells

    PubMed Central

    Xu, Hongsheng; Zong, Hailiang; Ma, Chong; Ming, Xing; Shang, Ming; Li, Kai; He, Xiaoguang; Cao, Lei

    2017-01-01

    The temperature variation rule of nano-magnetic fluid in the specific magnetic field and the effect on the treatment of malignant glioma were examined. The temperature variation of nano-magnetic fluid in the specific magnetic field was investigated by heating in vitro, and cell morphology was observed through optical microscopy and electron microscopy. MTT detection also was used to detect the effect of Fe3O4 nanometer magnetic fluid hyperthermia (MFH) on the proliferation of human U251 glioma cell line. The Fe3O4 nano MFH experiment was used to detect the inhibition rate of the tumor volume in nude mice with tumors. The results of the experiment showed that the heating ability of magnetic fluid was positively correlated with its concentration at the same intensity of the magnetic field. The results also indicated the prominent inhibitory effect of nanometer MFH on the proliferation of glioma cells, which was a dose-dependent relationship with nanometer magnetic fluid concentration. The hyperthermia experiment of nude mice with tumors displayed a significant inhibiting effect of Fe3O4 nanometer magnetic fluid in glioma volume. These results explain that iron (II, III) oxide (Fe3O4) nanometer MFH can inhibit the proliferation of U251 glioma cells, and has an obvious inhibitory effect on glioma volume, which plays a certain role in the treatment of brain glioma. PMID:28356945

  10. Evaluation of nano-magnetic fluid on malignant glioma cells.

    PubMed

    Xu, Hongsheng; Zong, Hailiang; Ma, Chong; Ming, Xing; Shang, Ming; Li, Kai; He, Xiaoguang; Cao, Lei

    2017-02-01

    The temperature variation rule of nano-magnetic fluid in the specific magnetic field and the effect on the treatment of malignant glioma were examined. The temperature variation of nano-magnetic fluid in the specific magnetic field was investigated by heating in vitro, and cell morphology was observed through optical microscopy and electron microscopy. MTT detection also was used to detect the effect of Fe3O4 nanometer magnetic fluid hyperthermia (MFH) on the proliferation of human U251 glioma cell line. The Fe3O4 nano MFH experiment was used to detect the inhibition rate of the tumor volume in nude mice with tumors. The results of the experiment showed that the heating ability of magnetic fluid was positively correlated with its concentration at the same intensity of the magnetic field. The results also indicated the prominent inhibitory effect of nanometer MFH on the proliferation of glioma cells, which was a dose-dependent relationship with nanometer magnetic fluid concentration. The hyperthermia experiment of nude mice with tumors displayed a significant inhibiting effect of Fe3O4 nanometer magnetic fluid in glioma volume. These results explain that iron (II, III) oxide (Fe3O4) nanometer MFH can inhibit the proliferation of U251 glioma cells, and has an obvious inhibitory effect on glioma volume, which plays a certain role in the treatment of brain glioma.

  11. COPD in HIV-Infected Patients: CD4 Cell Count Highly Correlated

    PubMed Central

    Guillouet-de-Salvador, Francine; Valerio, Laure; Puglièse, Pascal; Naqvi, Alissa; Durant, Jacques; Demonchy, Elisa; Perbost, Isabelle; Cua, Eric; Marquette, Charles-Hugo; Roger, Pierre-Marie

    2017-01-01

    Background COPD is a frequent and significant cause of respiratory morbidity in HIV-infected patients despite the control of HIV. We aimed to analyze the factors correlated with COPD in this population to evaluate the existence of specific indicators of vulnerability in this population. Methods and Findings 623 HIV-infected outpatients were enrolled during one year. This population was characterised by a dedicated questionnaire and electronic patient records. COPD screening was performed according to recommended spirometric criteria. The prevalence of COPD was 9.0%. Age and smoking were independently correlated with COPD (OR, 1.61 per 10 years increase, P = 0.007; OR, 1.28 per 10 pack-year increase, P = 0.003, respectively). Body mass index (BMI) and CD4 cell-count were independently and negatively correlated with COPD (OR, 0.78, P < 0.001; 0R, 0.77 per 100 cell/mm3 increase, P < 0.001, respectively). Among COPD patients, 77% did not know their diagnosis. Five COPD-patients never smoked and 44.2% did not have any respiratory symptoms and so were not eligible to perform a spirometry according to the guidelines. Conclusions In addition to known risk factors, immune defect through CD4 cell count was independently and strongly correlated with COPD. COPD is largely underdiagnosed and thus unmanaged. However, early management and urgent smoking cessation are essential to improve prognosis. Clinicians’ awareness on the particular vulnerability for COPD in HIV-infected patients is crucial. Moreover, indications to perform conventional spirometry to diagnose COPD may include more parameters than tobacco-smoking and respiratory complaints with a particular concern toward patients with a profound CD4 cell count defect. PMID:28056048

  12. Road transportation stress promptly increases bovine peripheral blood absolute NK cell counts and cortisol levels.

    PubMed

    Ishizaki, Hiroshi; Kariya, Yoshihiro

    2010-06-01

    Livestock transportation effects on the number of circulating leukocytes have been reported. However, data related specifically to the relation between acute stress levels during transport and leukocyte differentiation, including lymphocyte subsets, are lacking. This study was undertaken to evaluate the distribution of peripheral blood leukocyte differential counts, CD25+ lymphocytes and NK cells in calves subjected to truck transportation on different road types. Healthy Japanese Black calves were divided into three treatments: 1) those moved around in a mountainous area (Group M); 2) those moved around on flatland (Group F); and 3) those that were not transported (control). The plasma cortisol levels in Group M increased during transport. The increase was significantly higher at the end of transport than in the controls (P<0.05); a slight increase was noted in Group F. Total leukocytes and the neutrophil to lymphocyte ratio in Group M were elevated with neutrophilia at 2 hr post-transport (P<0.05); the former levels remained higher than those in the controls for 4 hr. The numbers of lymphocytes, monocytes, eosinophils and CD25+ lymphocytes remained unchanged throughout the observations. The number of circulating NK cells in Group M increased during transport and peaked shortly after transport (P<0.05). Subsequent to these time points, the counts in Group F showed a trend toward elevation. The circulating NK cell counts were positively correlated with the plasma cortisol level during transport (M, r=0.755, P<0.0005; F, r=0.653; P<0.005). These results suggest that circulating NK cells might be more rapidly mobilized than other leukocytes. Therefore, they might reflect acute stress levels in calves during road transportation.

  13. Usefulness of combined white blood cell count and plasma glucose for predicting in-hospital outcomes after acute myocardial infarction.

    PubMed

    Ishihara, Masaharu; Kojima, Sunao; Sakamoto, Tomohiro; Asada, Yujiro; Kimura, Kazuo; Miyazaki, Shunichi; Yamagishi, Masakazu; Tei, Chuwa; Hiraoka, Hisatoyo; Sonoda, Masahiro; Tsuchihashi, Kazufumi; Shinoyama, Nobuo; Honda, Takashi; Ogata, Yasuhiro; Ogawa, Hisao

    2006-06-01

    Admission white blood cell (WBC) count and plasma glucose (PG) have been associated with adverse outcomes after acute myocardial infarction (AMI). This study investigated the joint effect of WBC count and PG on predicting in-hospital outcomes in patients with AMI. WBC count and PG were measured at the time of hospital admission in 3,665 patients with AMI. Patients were stratified into tertiles (low, medium, and high) based on WBC count and PG. Patients with a high WBC count had a 2.0-fold increase in in-hospital mortality compared with those with a low WBC count. Patients with a high PG level had a 2.7-fold increase in mortality compared with those with a low PG level. When a combination of different strata for each variable was analyzed, a stepwise increase in mortality was seen. There was a considerable number of patients with a high WBC count and low PG level or with a low WBC count and high PG level. These patients had an intermediate risk, whereas those with a high WBC count and high PG level had the highest risk, i.e., 4.8-fold increase in mortality, compared with those with a low WBC count and low PG level. Multivariate analysis was performed to assess the predictor for in-hospital mortality using WBC count and PG level as continuous variables and showed that WBC count and PG level were independently associated with in-hospital mortality. These findings suggested that a simple combination of WBC count and PG level might provide further information for predicting outcomes in patients with AMI.

  14. A more appropriate white blood cell count for estimating malaria parasite density in Plasmodium vivax patients in northeastern Myanmar.

    PubMed

    Liu, Huaie; Feng, Guohua; Zeng, Weilin; Li, Xiaomei; Bai, Yao; Deng, Shuang; Ruan, Yonghua; Morris, James; Li, Siman; Yang, Zhaoqing; Cui, Liwang

    2016-04-01

    The conventional method of estimating parasite densities employ an assumption of 8000 white blood cells (WBCs)/μl. However, due to leucopenia in malaria patients, this number appears to overestimate parasite densities. In this study, we assessed the accuracy of parasite density estimated using this assumed WBC count in eastern Myanmar, where Plasmodium vivax has become increasingly prevalent. From 256 patients with uncomplicated P. vivax malaria, we estimated parasite density and counted WBCs by using an automated blood cell counter. It was found that WBC counts were not significantly different between patients of different gender, axillary temperature, and body mass index levels, whereas they were significantly different between age groups of patients and the time points of measurement. The median parasite densities calculated with the actual WBC counts (1903/μl) and the assumed WBC count of 8000/μl (2570/μl) were significantly different. We demonstrated that using the assumed WBC count of 8000 cells/μl to estimate parasite densities of P. vivax malaria patients in this area would lead to an overestimation. For P. vivax patients aged five years and older, an assumed WBC count of 5500/μl best estimated parasite densities. This study provides more realistic assumed WBC counts for estimating parasite densities in P. vivax patients from low-endemicity areas of Southeast Asia.

  15. Oligonol Supplementation Affects Leukocyte and Immune Cell Counts after Heat Loading in Humans

    PubMed Central

    Lee, Jeong Beom; Shin, Young Oh

    2014-01-01

    Oligonol is a low-molecular-weight form of polyphenol and has antioxidant and anti-inflammatory activity, making it a potential promoter of immunity. This study investigates the effects of oligonol supplementation on leukocyte and immune cell counts after heat loading in 19 healthy male volunteers. The participants took a daily dose of 200 mg oligonol or a placebo for 1 week. After a 2-week washout period, the subjects were switched to the other study arm. After each supplement, half-body immersion into hot water was made, and blood was collected. Then, complete and differential blood counts were performed. Flow cytometry was used to enumerate and phenotype lymphocyte subsets. Serum concentrations of interleukin (IL)-1β and IL-6 in blood samples were analyzed. Lymphocyte subpopulation variables included counts of total T cells, B cells, and natural killer (NK) cells. Oligonol intake attenuated elevations in IL-1β (an 11.1-fold change vs. a 13.9-fold change immediately after heating; a 12.0-fold change vs. a 12.6-fold change 1h after heating) and IL-6 (an 8.6-fold change vs. a 9.9-fold change immediately after heating; a 9.1-fold change vs. a 10.5-fold change 1h after heating) immediately and 1 h after heating in comparison to those in the placebo group. Oligonol supplementation led to significantly higher numbers of leukocytes (a 30.0% change vs. a 21.5% change immediately after heating; a 13.5% change vs. a 3.5% change 1h after heating) and lymphocytes (a 47.3% change vs. a 39.3% change immediately after heating; a 19.08% change vs. a 2.1% change 1h after heating) relative to those in the placebo group. Oligonol intake led to larger increases in T cells, B cells, and NK cells at rest (p < 0.05, p < 0.05, and p < 0.001, respectively) and immediately after heating (p < 0.001) in comparison to those in the placebo group. In addition, levels of T cells (p < 0.001) and B cells (p < 0.001) were significantly higher 1 h after heating in comparison to those in the

  16. Fluid shear, intercellular stress, and endothelial cell alignment.

    PubMed

    Steward, Robert; Tambe, Dhananjay; Hardin, C Corey; Krishnan, Ramaswamy; Fredberg, Jeffrey J

    2015-04-15

    Endothelial cell alignment along the direction of laminar fluid flow is widely understood to be a defining morphological feature of vascular homeostasis. While the role of associated signaling and structural events have been well studied, associated intercellular stresses under laminar fluid shear have remained ill-defined and the role of these stresses in the alignment process has remained obscure. To fill this gap, we report here the tractions as well as the complete in-plane intercellular stress fields measured within the human umbilical vein endothelial cell (HUVEC) monolayer subjected to a steady laminar fluid shear of 1 Pa. Tractions, intercellular stresses, as well as their time course, heterogeneity, and anisotropy, were measured using monolayer traction microscopy and monolayer stress microscopy. Prior to application of laminar fluid flow, intercellular stresses were largely tensile but fluctuated dramatically in space and in time (317 ± 122 Pa). Within 12 h of the onset of laminar fluid flow, the intercellular stresses decreased substantially but continued to fluctuate dramatically (142 ± 84 Pa). Moreover, tractions and intercellular stresses aligned strongly and promptly (within 1 h) along the direction of fluid flow, whereas the endothelial cell body aligned less strongly and substantially more slowly (12 h). Taken together, these results reveal that steady laminar fluid flow induces prompt reduction in magnitude and alignment of tractions and intercellular stress tensor components followed by the retarded elongation and alignment of the endothelial cell body. Appreciably smaller intercellular stresses supported by cell-cell junctions logically favor smaller incidence of gap formation and thus improved barrier integrity.

  17. Cell sorting is analogous to phase ordering in fluids

    PubMed Central

    Beysens, D. A.; Forgacs, G.; Glazier, J. A.

    2000-01-01

    Morphogenetic processes, like sorting or spreading of tissues, characterize early embryonic development. An analogy between viscoelastic fluids and certain properties of embryonic tissues helps interpret these phenomena. The values of tissue-specific surface tensions are consistent with the equilibrium configurations that the Differential Adhesion Hypothesis predicts such tissues reach after sorting and spreading. Here we extend the fluid analogy to cellular kinetics. The same formalism applies to recent experiments on the kinetics of phase ordering in two-phase fluids. Our results provide biologically relevant information on the strength of binding between cell adhesion molecules under near-physiological conditions. PMID:10944216

  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.

  19. Neurocognitive and Motor Deficits in HIV-Infected Ugandan Children With High CD4 Cell Counts

    PubMed Central

    Boivin, Michael J.; Boal, Hannah E.; Bangirana, Paul; Charlebois, Edwin; Havlir, Diane V.; Rosenthal, Philip J.; Dorsey, Grant; Achan, Jane; Akello, Carolyne; Kamya, Moses R.; Wong, Joseph K.

    2012-01-01

    (See the Editorial Commentary by Wagner and Frenkel, on pages 1010–2.) Background. Human immunodeficiency virus (HIV) infection causes neurocognitive or motor function deficits in children with advanced disease, but it is unclear whether children with CD4 cell measures above the World Health Organization (WHO) thresholds for antiretroviral therapy (ART) initiation suffer significant impairment. Methods. The neurocognitive and motor functions of HIV-infected ART-naive Ugandan children aged 6–12 years with CD4 cell counts of >350 cells/μL and CD4 cell percentage of >15% were compared with those of HIV-uninfected children, using the Test of Variables of Attention (TOVA), the Kaufman Assessment Battery for Children, second edition (KABC-2), and the Bruininks-Oseretsky Test of Motor Proficiency, second edition (BOT-2). Results. Ninety-three HIV-infected children (median CD4 cell count, 655 cells/μL; plasma HIV RNA level, 4.7 log10 copies/mL) were compared to 106 HIV-uninfected children. HIV-infected children performed worse on TOVA visual reaction times (multivariate analysis of covariance; P = .006); KABC-2 sequential processing (P = .005), simultaneous processing (P = .039), planning/reasoning (P = .023), and global performance (P = .024); and BOT-2 total motor proficiency (P = .003). High plasma HIV RNA level was associated with worse performance in 10 cognitive measures and 3 motor measures. In analysis of only WHO clinical stage 1 or 2 HIV-infected children (n = 68), significant differences between the HIV-infected and HIV-uninfected groups (P < .05) remained for KABC-2 sequential processing, KABC-2 planning/reasoning, and BOT-2 motor proficiency. Conclusions. Significant motor and cognitive deficits were found in HIV-infected ART-naive Ugandan children with CD4 cell counts of ∼350 cells/μL and percentages of >15%. Study of whether early initiation of ART could prevent or reverse such deficits is needed. PMID

  20. A Multiple Parameters Biodosimetry Tool with Various Blood Cell Counts - the Hemodose Approach

    NASA Technical Reports Server (NTRS)

    Hu, Shaowen

    2014-01-01

    There continue to be important concerns about the possibility of the occurrence of acute radiation syndromes following nuclear and radiological terrorism or accidents that may result in mass casualties in densely populated areas. To guide medical personnel in their clinical decisions for effective medical management and treatment of the exposed individuals, biological markers are usually applied to examine radiation induced biological changes to assess the severity of radiation injury to sensitive organ systems. Among these the peripheral blood cell counts are widely used to assess the extent of radiation induced bone marrow injury. This is due to the fact that the hematopoietic system is the most vulnerable part of the human body to radiation damage. Particularly, the lymphocyte, granulocyte, and platelet cells are the most radiosensitive of the blood elements, and monitoring their changes after exposure is regarded as a practical and recommended laboratory test to estimate radiation dose and injury. Based upon years of physiological and pathophysiological investigation of mammalian hematopoietic systems, and rigorous coarse-grained bio-mathematical modeling and validation on species from mouse, to dog, monkey, and human, we have developed a set of software tools Hemodose, which can use single or serial granulocyte, lymphocyte, leukocyte, or platelet counts after exposure to estimate absorbed doses of adult victims very rapidly and accurately. Some patient data from historical accidents are utilized as examples to demonstrate the capabilities of these tools as a rapid point-of-care diagnostic or centralized high-throughput assay system in a large-scale radiological disaster scenario. Most significant to the improvement of national and local preparedness of a potential nuclear/radiological disaster, this HemoDose approach establishes robust correlations between the absorbed doses and victim's various types of blood cell counts not only in the early time window (1

  1. A robust generic method for grid detection in white light microscopy Malassez blade images in the context of cell counting.

    PubMed

    Marin, Ambroise; Denimal, Emmanuel; Guyot, Stéphane; Journaux, Ludovic; Molin, Paul

    2015-02-01

    In biology, cell counting is a primary measurement and it is usually performed manually using hemocytometers such as Malassez blades. This work is tedious and can be automated using image processing. An algorithm based on Fourier transform filtering and the Hough transform was developed for Malassez blade grid extraction. This facilitates cell segmentation and counting within the grid. For the present work, a set of 137 images with high variability was processed. Grids were accurately detected in 98% of these images.

  2. 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.

  3. Exogenous melatonin reduces somatic cell count of milk in Holstein cows.

    PubMed

    Yang, Minghui; Shi, Jianmin; Tian, Jianhua; Tao, Jingli; Chai, Menglong; Wang, Jing; Xu, Zhiyuan; Song, Yukun; Zhu, Kuanfeng; Ji, Pengyun; Liu, Guoshi

    2017-02-27

    High somatic cell counts in milk caused by mastitis significantly influence the quality of milk and result in substantial annual economic loss. This study evaluated the beneficial effects of melatonin (MT) on milk somatic cell count (SCC) in cows. To examine the effects of melatonin on SCC, one hundred twenty cows were divided into four groups based on milk SCC. In each group, half of the cows were treated with melatonin (S.C.). Melatonin treatment significantly reduced milk SCC. To explore the potential mechanism, 20 cows with relatively high SCC were selected to evaluate the biochemical and immunological profiles of their blood after melatonin treatment. Treatment with MT significantly reduced SCC in milk, lowered serum cortisol concentrations and increased the levels of albumin, alanine transaminase and lactate dehydrogenase. Following treatment with MT, the concentration of IgG and IgM rose transiently then decreased significantly, similar to changes observed for white blood cells and lymphocytes. In conclusion, MT treatment improved the quality of milk by reducing SCC. This may be due to melatonin improving immune activity in cows.

  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. Telomere Length in Elderly Caucasians Weakly Correlates with Blood Cell Counts

    PubMed Central

    Witecka, Joanna; Koscinska-Marczewska, Justyna; Szwed, Malgorzata; Owczarz, Magdalena; Mossakowska, Malgorzata; Milewicz, Andrzej; Zejda, Jan; Wiecek, Andrzej

    2013-01-01

    Background. Age-related decrease in bone marrow erythropoietic capacity is often accompanied by the telomere length shortening in peripheral white blood cells. However, limited and conflicting data hamper the conclusive opinion regarding this relationship. Therefore, the aim of this study was to assess an association between telomere length and peripheral blood cell count parameters in the Polish elderly population. Material and Methods. The substudy included 1573 of 4981 subjects aged 65 years or over, participants of the population-based PolSenior study. High-molecular-weight DNA was isolated from blood mononuclear cells. Telomere length (TL) was measured by QRT-PCR as abundance of telomere template versus a single gene copy encoding acidic ribosomal phosphoprotein P0. Results. Only white blood count (WBC) was significantly different in TL tertile subgroups in all subjects (P = 0.02) and in men (P = 0.01), but not in women. Merely in men significant but weak positive correlations were found between TL and WBC (r = 0.11, P < 0.05) and RBC (r = 0.08, P < 0.05). The multiple regression analysis models confirmed a weak, independent contribution of TL to both RBC and WBC. Conclusions. In the elderly, telomere shortening limits hematopoiesis capacity to a very limited extent. PMID:24453794

  6. Automated counting of morphologically normal red blood cells by using digital holographic microscopy and statistical methods

    NASA Astrophysics Data System (ADS)

    Moon, Inkyu; Yi, Faliu

    2015-09-01

    In this paper we overview a method to automatically count morphologically normal red blood cells (RBCs) by using off-axis digital holographic microscopy and statistical methods. Three kinds of RBC are used as training and testing data. All of the RBC phase images are obtained with digital holographic microscopy (DHM) that is robust to transparent or semitransparent biological cells. For the determination of morphologically normal RBCs, the RBC's phase images are first segmented with marker-controlled watershed transform algorithm. Multiple features are extracted from the segmented cells. Moreover, the statistical method of Hotelling's T-square test is conducted to show that the 3D features from 3D imaging method can improve the discrimination performance for counting of normal shapes of RBCs. Finally, the classifier is designed by using statistical Bayesian algorithm and the misclassification rates are measured with leave-one-out technique. Experimental results show the feasibility of the classification method for calculating the percentage of each typical normal RBC shape.

  7. Exogenous melatonin reduces somatic cell count of milk in Holstein cows

    PubMed Central

    Yang, Minghui; Shi, Jianmin; Tian, Jianhua; Tao, Jingli; Chai, Menglong; Wang, Jing; Xu, Zhiyuan; Song, Yukun; Zhu, Kuanfeng; Ji, Pengyun; Liu, Guoshi

    2017-01-01

    High somatic cell counts in milk caused by mastitis significantly influence the quality of milk and result in substantial annual economic loss. This study evaluated the beneficial effects of melatonin (MT) on milk somatic cell count (SCC) in cows. To examine the effects of melatonin on SCC, one hundred twenty cows were divided into four groups based on milk SCC. In each group, half of the cows were treated with melatonin (S.C.). Melatonin treatment significantly reduced milk SCC. To explore the potential mechanism, 20 cows with relatively high SCC were selected to evaluate the biochemical and immunological profiles of their blood after melatonin treatment. Treatment with MT significantly reduced SCC in milk, lowered serum cortisol concentrations and increased the levels of albumin, alanine transaminase and lactate dehydrogenase. Following treatment with MT, the concentration of IgG and IgM rose transiently then decreased significantly, similar to changes observed for white blood cells and lymphocytes. In conclusion, MT treatment improved the quality of milk by reducing SCC. This may be due to melatonin improving immune activity in cows. PMID:28240296

  8. Monitoring the biomass accumulation of recombinant yeast cultures: offline estimations of dry cell mass and cell counts.

    PubMed

    Palmer, Shane M; Kunji, Edmund R S

    2012-01-01

    Biomass is one of the most important parameters for process optimization, scale-up and control in recombinant protein production experiments. However, a standard unit of biomass remains elusive. Methods of biomass monitoring have increasingly been developed towards online, in situ techniques in order to advance process analysis and control. Offline, ex situ methods, such as dry cell mass determination and direct cell counts, remain the reference for determining cell mass and number, respectively, but this type of analysis is time consuming. In this chapter, protocols are presented for determining these offline measures of the biomass yield of recombinant yeast cultures.

  9. Urinary CD8+ T-cell counts discriminate between active and inactive lupus nephritis

    PubMed Central

    2013-01-01

    Introduction Lupus nephritis (LN) is a severe and frequent manifestation of systemic lupus erythematosus (SLE). Early detection of initial renal manifestations and relapses during follow-up is pivotal to prevent loss of renal function. Apart from renal biopsies, current urinary and serological diagnostic tests fail to accurately demonstrate the presence of active LN. Previously, we demonstrated that effector memory T-cells (CD45RO+CCR7-;TEM) migrate into the urine during active LN. The objective of this study was to assess the diagnostic value of urinary T-cells in comparison with traditional markers of active LN. Methods T-cells in the urine during active LN and remission were investigated. Twenty-two, in most cases biopsy-proven, active LN patients and 24 SLE patients without active LN were enrolled and serial measurements were performed in 16 patients. Results Analysis of the urinary sediment in active renal disease showed an increased number of CD8+ T-cells and absence of these cells during remission. Enumerating T-cell counts in LN patients with a history of renal involvement was a superior marker of active LN in comparison to traditional markers, such as proteinuria and s-creatinine. Conclusions In conclusion, urinary T-cells, in particular CD8+ T cells, are a promising marker to assess renal activity in LN patients, in particular in those with prior renal involvement. PMID:23445537

  10. Method of detecting and counting bacteria

    NASA Technical Reports Server (NTRS)

    Picciolo, G. L.; Chappelle, E. W. (Inventor)

    1976-01-01

    An improved method is provided for determining bacterial levels, especially in samples of aqueous physiological fluids. The method depends on the quantitative determination of bacterial adenosine triphosphate (ATP) in the presence of nonbacterial ATP. The bacterial ATP is released by cell rupture and is measured by an enzymatic bioluminescent assay. A concentration technique is included to make the method more sensitive. It is particularly useful where the fluid to be measured contains an unknown or low bacteria count.

  11. Action spectrum for bergamot-oil phototoxicity measured by sunburn cell counting.

    PubMed

    Yasui, Y; Hirone, T

    1994-05-01

    The present study investigated the phototoxic effect of bergamot oil and its photosensitive component, bergapten, on sunburn cell (SBC) production in guinea pig skin. The back skin was pretreated with bergamot oil or bergapten and exposed to monochromatic light under various conditions. After irradiation, skin specimens were excised, and histological sections were prepared. The number of sunburn cells in the interfollicular epidermis was counted. The SBC formation by bergamot oil or bergapten plus UVB radiation was the same as that without pretreatment with any photosensitizer. In contrast, a significant number of SBCs were induced by bergamot oil or bergapten plus UVA radiation, but no SBCs were found after the treatment with UVA alone. The result indicates that bergamot oil or bergapten was photosensitized by UVA irradiation. The SBCs were linearly increased in a UV-dose dependent manner. On the basis of the regression lines, an action spectrum and spectral peak for the photosensitizers plus UVA were obtained. The action spectrum for bergamot oil- and bergapten-induced SBC formation was in the ranges of 325-365 nm and 325-350 nm, and their spectral peaks were at 335-345 nm and 335-350 nm, respectively. The data are in good accordance with those estimated from skin erythema reactions. Therefore, counting SBCs is a very useful parameter for quantitative evaluation of phototoxicity.

  12. Short communication: contribution of vibration and noise during milking to the somatic cell count of milk.

    PubMed

    Gygax, L; Nosal, D

    2006-07-01

    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 increasing intensity of vibration but not with acoustic noise. Cows milked in autotandem and side-by-side parlors had lower SCC than those in the other 2 types of milking parlors. On 12 farms where the milking system was modified to reduce vibration and noise, SCC also dropped. In addition, the relative improvement in SCC seemed to be correlated with the relative improvement in the reduction of vibration but not with the improvement in acoustic noise. A reduction in vibration (structure-borne sonic waves) seemed to improve udder health, which may have been mediated by the increased well-being and reduced stress of cows during milking.

  13. Low blood cell counts in wild Japanese monkeys after the Fukushima Daiichi nuclear disaster.

    PubMed

    Ochiai, Kazuhiko; Hayama, Shin-ichi; Nakiri, Sachie; Nakanishi, Setsuko; Ishii, Naomi; Uno, Taiki; Kato, Takuya; Konno, Fumiharu; Kawamoto, Yoshi; Tsuchida, Shuichi; Omi, Toshinori

    2014-07-24

    In April 2012 we carried out a 1-year hematological study on a population of wild Japanese monkeys inhabiting the forest area of Fukushima City. This area is located 70 km from the Fukushima Daiichi Nuclear Power Plant (NPP), which released a large amount of radioactive material into the environment following the Great East Japan Earthquake of 2011. For comparison, we examined monkeys inhabiting the Shimokita Peninsula in Aomori Prefecture, located approximately 400 km from the NPP. Total muscle cesium concentration in Fukushima monkeys was in the range of 78-1778 Bq/kg, whereas the level of cesium was below the detection limit in all Shimokita monkeys. Compared with Shimokita monkeys, Fukushima monkeys had significantly low white and red blood cell counts, hemoglobin, and hematocrit, and the white blood cell count in immature monkeys showed a significant negative correlation with muscle cesium concentration. These results suggest that the exposure to some form of radioactive material contributed to hematological changes in Fukushima monkeys.

  14. Low blood cell counts in wild Japanese monkeys after the Fukushima Daiichi nuclear disaster

    PubMed Central

    Ochiai, Kazuhiko; Hayama, Shin-ichi; Nakiri, Sachie; Nakanishi, Setsuko; Ishii, Naomi; Uno, Taiki; Kato, Takuya; Konno, Fumiharu; Kawamoto, Yoshi; Tsuchida, Shuichi; Omi, Toshinori

    2014-01-01

    In April 2012 we carried out a 1-year hematological study on a population of wild Japanese monkeys inhabiting the forest area of Fukushima City. This area is located 70 km from the Fukushima Daiichi Nuclear Power Plant (NPP), which released a large amount of radioactive material into the environment following the Great East Japan Earthquake of 2011. For comparison, we examined monkeys inhabiting the Shimokita Peninsula in Aomori Prefecture, located approximately 400 km from the NPP. Total muscle cesium concentration in Fukushima monkeys was in the range of 78–1778 Bq/kg, whereas the level of cesium was below the detection limit in all Shimokita monkeys. Compared with Shimokita monkeys, Fukushima monkeys had significantly low white and red blood cell counts, hemoglobin, and hematocrit, and the white blood cell count in immature monkeys showed a significant negative correlation with muscle cesium concentration. These results suggest that the exposure to some form of radioactive material contributed to hematological changes in Fukushima monkeys. PMID:25060710

  15. Amniotic fluid-derived stem cells in regenerative medicine research.

    PubMed

    Joo, Sunyoung; Ko, In Kap; Atala, Anthony; Yoo, James J; Lee, Sang Jin

    2012-02-01

    The stem cells isolated from amniotic fluid present an exciting possible contribution to the field of regenerative medicine and amniotic fluid-derived stem (AFS) cells have significant potential for research and therapeutic applications. AFS cells are multipotent, showing the ability to differentiate into cell types from all three embryonic germ layers. They express both embryonic and adult stem cell markers, expand extensively without feeder cells, double in 36 h, and are not tumorigenic. The AFS cells can be maintained for over 250 population doublings and preserve their telomere length and a normal karyotype. They differentiate easily into specific cell lineages and do not require human embryo tissue for their isolation, thus avoiding the current controversies associated with the use of human embryonic stem (ES) cells. The discovery of the AFS cells has been recent, and a great deal of work remains to be performed on the characterization and use of these cells. This review describes the various differentiated lineages that AFS cells can form and the future of these promising new stem cells in regenerative medicine research.

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

    PubMed

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

    2014-08-01

    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.

  17. A new approach for the estimation of phytoplankton cell counts associated with algal blooms.

    PubMed

    Nazeer, Majid; Wong, Man Sing; Nichol, Janet Elizabeth

    2017-07-15

    This study proposes a method for estimating phytoplankton cell counts associated with an algal bloom, using satellite images coincident with in situ and meteorological parameters. Satellite images from Landsat Thematic Mapper (TM), Enhanced Thematic Mapper Plus (ETM+), Operational Land Imager (OLI) and HJ-1 A/B Charge Couple Device (CCD) sensors were integrated with the meteorological observations to provide an estimate of phytoplankton cell counts. All images were atmospherically corrected using the Second Simulation of the Satellite Signal in the Solar Spectrum (6S) atmospheric correction method with a possible error of 1.2%, 2.6%, 1.4% and 2.3% for blue (450-520nm), green (520-600nm), red (630-690nm) and near infrared (NIR 760-900nm) wavelengths, respectively. Results showed that the developed Artificial Neural Network (ANN) model yields a correlation coefficient (R) of 0.95 with the in situ validation data with Sum of Squared Error (SSE) of 0.34cell/ml, Mean Relative Error (MRE) of 0.154cells/ml and a bias of -504.87. The integration of the meteorological parameters with remote sensing observations provided a promising estimation of the algal scum as compared to previous studies. The applicability of the ANN model was tested over Hong Kong as well as over Lake Kasumigaura, Japan and Lake Okeechobee, Florida USA, where algal blooms were also reported. Further, a 40-year (1975-2014) red tide occurrence map was developed and revealed that the eastern and southern waters of Hong Kong are more vulnerable to red tides. Over the 40 years, 66% of red tide incidents were associated with the Dinoflagellates group, while the remainder were associated with the Diatom group (14%) and several other minor groups (20%). The developed technology can be applied to other similar environments in an efficient and cost-saving manner.

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

    PubMed

    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.

  19. Prognostic value of parameters derived from white blood cell and differential counts in patients receiving palliative radiotherapy.

    PubMed

    Saito, Tetsuo; Toya, Ryo; Matsuyama, Tomohiko; Semba, Akiko; Matsuyama, Keiya; Oya, Natsuo

    2016-09-01

    The aim of the present study was to identify white blood cell (WBC) parameters with high prognostic value for the survival of patients receiving palliative radiotherapy. The prognostic value of seven parameters derived from WBC and differential counts was retrospectively evaluated in patients who underwent palliative radiotherapy between October, 2010 and June, 2013. The analyzed parameters were the total WBC count, the absolute and relative lymphocyte count, the absolute and relative neutrophil count, and the neutrophil-to-lymphocyte and lymphocyte-to-monocyte ratios. Following univariate analysis, multivariate Cox regression analysis was performed to adjust for gender, age, disease type, previous chemotherapy, previous radiotherapy and the levels of albumin and lactate dehydrogenase. A total of 220 patients with a median survival of 4.7 months were identified. All seven parameters were found to be statistically significant predictors of survival on univariate Cox regression analysis (P<0.05). Of these parameters, the low relative lymphocyte and high relative neutrophil counts were consistent predictors of poor survival in patients who received chemotherapy within 1 month prior to blood sampling (n=68) and in patients who received steroid treatment at the time of sampling (n=49). Multivariate Cox regression analysis revealed that the relative lymphocyte and neutrophil counts were independent predictors of survival in all 220 patients (P<0.05). In conclusion, relative lymphocyte and neutrophil counts were of high prognostic value for the survival of patients receiving palliative radiotherapy, even in those receiving medications that affect WBC and differential counts.

  20. Alternative experiments using the geophysical fluid flow cell

    NASA Technical Reports Server (NTRS)

    Hart, J. E.

    1984-01-01

    This study addresses the possibility of doing large scale dynamics experiments using the Geophysical Fluid Flow Cell. In particular, cases where the forcing generates a statically stable stratification almost everywhere in the spherical shell are evaluated. This situation is typical of the Earth's atmosphere and oceans. By calculating the strongest meridional circulation expected in the spacelab experiments, and testing its stability using quasi-geostrophic stability theory, it is shown that strongly nonlinear baroclinic waves on a zonally symmetric modified thermal wind will not occur. The Geophysical Fluid Flow Cell does not have a deep enough fluid layer to permit useful studies of large scale planetary wave processes arising from instability. It is argued, however, that by introducing suitable meridional barriers, a significant contribution to the understanding of the oceanic thermocline problem could be made.

  1. Amniotic fluid stem cells increase embryo survival following injury.

    PubMed

    Prasongchean, Weerapong; Bagni, Marinella; Calzarossa, Cinzia; De Coppi, Paolo; Ferretti, Patrizia

    2012-03-20

    Although amniotic fluid cells can differentiate into several mesenchymal lineages and have been proposed as a valuable therapeutic cell source, their ability to undergo terminal neuronal differentiation remains a cause of controversy. The aim of this study was to investigate the neuronal differentiation ability of the c-Kit-positive population from GFP-transgenic rat amniotic fluid, amniotic fluid stem (AFS) cells, and to assess how they affected injury response in avian embryos. AFS cells were found to express several neural stem/progenitor cell markers. However, no overt neuronal differentiation was apparent after either treatment with small molecules known to stimulate neuronal differentiation, attempts to differentiate AFS cell-derived embryoid body-like structures, or grafting AFS cells into environments known to support neuronal differentiation (organotypic rat hippocampal cultures, embryonic chick nervous system). Nonetheless, AFS cells significantly reduced hemorrhage and increased survival when grafted at the site of an extensive thoracic crush injury in E2.5 chick embryos. Increased embryo survival was induced neither by desmopressin treatment, which also reduced hemorrhage, nor by grafting other mesenchymal or neural cells, indicating a specific effect of AFS cells. This was found to be mediated by soluble factors in a transwell coculture model. Altogether, this study shows that AFS cells reduce tissue damage and increase survival in injured embryos, providing a potentially valuable tool as therapeutic agents for tissue repair, particularly prenatal/perinatal repair of defects diagnosed during gestation, but this effect is mediated via paracrine mechanisms rather than the ability of AFS cells to fully differentiate into neuronal cells.

  2. 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.

  3. 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

  4. Evaluation of the use of dry cow antibiotics in low somatic cell count cows.

    PubMed

    Scherpenzeel, C G M; den Uijl, I E M; van Schaik, G; Olde Riekerink, R G M; Keurentjes, J M; Lam, T J G M

    2014-01-01

    The goal of dry cow therapy (DCT) is to reduce the prevalence of intramammary infections (IMI) by eliminating existing IMI at drying off and preventing new IMI from occurring during the dry period. Due to public health concerns, however, preventive use of antibiotics has become questionable. This study evaluated selective DCT in 1,657 cows with low somatic cell count (SCC) at the last milk recording before drying off in 97 Dutch dairy herds. Low SCC was defined as <150,000 cells/mL for primiparous and <250,000 cells/mL for multiparous cows. A split-udder design was used in which 2 quarters of each cow were treated with dry cow antibiotics and the other 2 quarters remained as untreated controls. The effect of DCT on clinical mastitis (CM), bacteriological status, SCC, and antibiotic use were determined at the quarter level using logistic regression and chi-squared tests. The incidence rate of CM was found to be 1.7 times (95% confidence interval = 1.4-2.1) higher in quarters dried off without antibiotics as compared with quarters dried off with antibiotics. Streptococcus uberis was the predominant organism causing CM in both groups. Somatic cell count at calving and 14 d in milk was significantly higher in quarters dried off without antibiotics (772,000 and 46,000 cells/mL, respectively) as compared with the quarters dried off with antibiotics (578,000 and 30,000 cells/mL, respectively). Quarters with an elevated SCC at drying off and quarters with a positive culture for major pathogens at drying off had a higher risk for an SCC above 200,000 cells/mL at 14 d in milk as compared with quarters with a low SCC at drying off and quarters with a negative culture for major pathogens at drying off. For quarters that were culture-positive for major pathogens at drying off, a trend for a higher risk on CM was also found. Selective DCT, not using DCT in cows that had a low SCC at the last milk recording before drying off, significantly increased the incidence rate of CM and

  5. Design of an Automated-Counting System of Cell Micronuclei in Micrographs

    NASA Astrophysics Data System (ADS)

    Lozano, A. V.; Márquez, J. A.; Buenfil, A. E.; Gonsebatt, M. E.

    2004-09-01

    We developed and tested a system for the automatic analysis of cell images in order to identify and count micronuclei configurations in digital images from a microscope. The presence of micronuclei has been used as an indicator of DNA damage, and a fast automated analysis may be suitable for early cancer detection, for example. We describe in this work the image-processing protocol and system comprising: acquisition, color normalization, contrast enhancement, color-background removing, color segmentation, mathematical-morphology filtering and restoration, morphometry, feature extraction and analysis, and counting. Among the morphological features used for discriminating micronuclei configurations, we tested compactness, area ratios and separation of selected features (ideally, a nucleus and one or more micronuclei). Among the mathematical-morphology techniques, we used for instance a modified watershed segmentation algorithm for separating touching features. We present our results on several images, the evaluation and performance of each step, and the main problems we solved by using several techniques from mathematical morphology and color-image processing.

  6. Monoclonal B-cell lymphocytosis (MBL) with normal lymphocyte counts is associated with decreased numbers of normal circulating B-cell subsets.

    PubMed

    Hauswirth, Alexander W; Almeida, Julia; Nieto, Wendy G; Teodosio, Cristina; Rodriguez-Caballero, Arancha; Romero, Alfonso; López, Antonio; Fernandez-Navarro, Paulino; Vega, Tomas; Perez-Andres, Martin; Valent, Peter; Jäger, Ulrich; Orfao, Alberto

    2012-07-01

    Monoclonal B-cell lymphocytosis (MBL) with normal lymphocyte counts is associated with decreased numbers of normal circulating B-cell subsets.Little is known about the distribution of normal lymphoid cells and their subsets in the peripheral blood (PB) of subjects with monoclonal B-cell lymphocytosis (MBL). In our study, we compared the absolute number of PB lymphoid cells and their subpopulations in 95 MBL cases with normal lymphocyte counts vs. 617 age-/sex-matched non-MBL healthy subjects (controls), using highly sensitive flow cytometry. MBL cases showed significantly reduced numbers of normal circulating B-cells, at the expense of immature and naive B-cells; in addition, CD4+CD8+ double-positive T-cells and CD8+ T-cells were significantly lower and higher vs. controls, respectively. Moreover, most normal B-cell subsets were significantly decreased in PB at >1% MBL-counts, vs. "low-count" MBL cases, and lower amounts of immature/naive B-cells were detected in biclonal (particularly in cases with coexisting CLL-like- and non-CLL-like B-cell clones) vs. monoclonal MBL subjects. In summary, our results show imbalanced (reduced) absolute numbers of recently produced normal circulating B-cells (e.g., immature and naıve B-cells) in MBL, which becomes more pronounced as the MBL cell count increases.

  7. Low Circulating Natural Killer Cell Counts are Associated With Severe Disease in Patients With Common Variable Immunodeficiency

    PubMed Central

    Ebbo, Mikael; Gérard, Laurence; Carpentier, Sabrina; Vély, Frédéric; Cypowyj, Sophie; Farnarier, Catherine; Vince, Nicolas; Malphettes, Marion; Fieschi, Claire; Oksenhendler, Eric; Schleinitz, Nicolas; Vivier, Eric

    2016-01-01

    Natural Killer (NK) cells have been shown to exert antiviral and antitumoural activities. Nevertheless most available data are derived from mouse models and functions of these cells in human remain unclear. To evaluate the impact of low circulating NK cell counts and to provide some clues to the role of NK cells in natural conditions, we studied a large cohort of patients with common variable immunodeficiency (CVID) included in a multicenter cohort of patients with primary hypogammaglobulinaemia. Patients were classified into three groups on the basis of their NK cell counts: severe and mild NK cell lymphopenia (< 50 and 50–99 × 106/L respectively), and normal NK cell counts (> 100 × 106/L). Clinical events were analyzed and compared between these three groups of patients. During study period, 457 CVID patients were included: 99 (21.7%) with severe NK cell lymphopenia, 118 (25.8%) with mild NK cell lymphopenia and 240 (52.5%) with normal NK cell counts. Non-infectious complications (57% vs. 36% and 35%), and, particularly, granulomatous complications (25.3% vs. 13.6% and 8.8%), were more frequent in patients with severe NK cell lymphopenia than in other groups. Invasive infections (68.7% vs. 60.2% and 48.8%), including bacteraemia (22.2% vs. 5.9% and 8.3%) and infectious pneumonia (63.6% vs. 59.3% and 44.2%), were also more frequent in this population. However, no difference was observed for viral infections and neoplasms. Low circulating NK cell counts are associated with more severe phenotypes of CVID, which may indicate a protective role of these immune cells against severe bacterial infections and other complications and non-redundant immune functions when the adaptive immune response is not optimal. PMID:27211564

  8. Analytic performance of the iQ200 automated urine microscopy analyzer and comparison with manual counts using Fuchs-Rosenthal cell chambers.

    PubMed

    Wah, David T; Wises, Porntip K; Butch, Anthony W

    2005-02-01

    Automated instruments that can examine urine for cells and particles have reduced the need for labor-intensive manual microscopy. We evaluated the performance of the Iris iQ200 Automated Urine Microscopy Analyzer (Iris Diagnostics, Chatsworth, CA) and compared results with manual cell and particle counts using Fuchs-Rosenthal counting chambers. Within-run imprecision (coefficient of variation) of the iQ200 for urine samples ranged from 3.0% to 45% for RBC counts between 1,029 and 3 x 10(6) cells/L, 3.4% to 40% for WBC counts between 1,006 and 4 x 10(6) cells/L, and 8.9% to 35% for epithelial cell counts between 93 and 4 x 10(6) cells/L. Between-run imprecision was 3.3% at 1,017 x 10(6) cells/L and 19.2% at 28 x 10(6) cells/L. There was good agreement between the iQ200 and manual cell counts (r > or = 0.94); however; the iQ200 produced lower results based on slopes of 0.92 (RBC count), 0.81 (WBC count), and 0.94 (epithelial cell counts). The iQ200 has satisfactory performance and correlates well with manual cell counts. Most urine samples containing RBCs, WBCs, and epithelial cells can be reported without review of captured images.

  9. Preoperative lymphocyte count is an independent prognostic factor in node-negative non-small cell lung cancer.

    PubMed

    Kobayashi, Naohiro; Usui, Shingo; Kikuchi, Shinji; Goto, Yukinobu; Sakai, Mitsuaki; Onizuka, Masataka; Sato, Yukio

    2012-02-01

    A number of prognostic factors have been reported in non-small cell lung cancer (NSCLC). Although lymph node metastasis is the most poorly predictive value in completely resected NSCLC, a significant number of patients have a fatal recurrence even in node-negative curative NSCLC. Recently inflammatory response has been shown as a predictive value in NSCLC. Neutrophils and lymphocytes play an important role in cancer immune response. In this study, we retrospectively examined the impact of preoperative peripheral neutrophil and lymphocyte counts on survival, and investigated the relationships of these factors to clinicopathological factors in node-negative NSCLC. A total 237 patients were evaluated. When the cut-off value of neutrophil count was 4500 mm(-3) with a maximum log-rank statistical value, overall 5-year survival rates were 79.7% for the low-neutrophil-count group and 69.5% for the high-neutrophil-count group (P=0.04). When the cut-off value of lymphocyte count was 1900 mm(-3) with a maximum log-rank statistical value, overall survival rates were 67.9% for the low-lymphocyte group and 87.7% for the high-lymphocyte group (P<0.001). High-neutrophil-counts were associated with tumor size (P=0.002) and pleural invasion (P<0.001). Low-lymphocyte-counts were correlated with vascular invasion (P=0.018) and recurrence of NSCLC (P=0.01). Multivariate analysis showed that the lymphocyte count was an independent prognostic factor (hazard ratio: 3.842; 95% confidence interval: 1.827-8.078; P<0.001), but the neutrophil count was not (P=0.185). We conclude that a peripheral lymphocyte count, which is associated with vascular invasion, is an independent prognostic factor in node-negative NCSLC.

  10. Intrauterine growth retardation in preterm infants ≤32 weeks of gestation is associated with low white blood cell counts.

    PubMed

    Wirbelauer, Johannes; Thomas, Wolfgang; Rieger, Lorenz; Speer, Christian P

    2010-11-01

    It is unclear if very immature preterm infants who are born small for gestational age (SGA) have similar leukocyte counts as infants who are born appropriate for gestational age (AGA). Our study included 49 preterm infants with a gestational age ≤32 weeks and without exposure to chorioamnionitis and funisitis. Blood cells were counted in the first 2 hours of life. Eighteen SGA preterm infants were compared with 31 AGA preterm infants. Gestational age, sex, rate of caesarean section, and prenatal administration of corticosteroids did not differ between the groups. Median birth weight was 583 g in the SGA group versus 1100 g in the AGA group. Infants in the SGA group had significantly lower counts of leukocytes, total neutrophils, immature neutrophils, lymphocytes, and monocytes. These findings were not affected by maternal preeclampsia. No significant difference for nucleated red blood cell counts was found. Prenatal growth retardation is an independent factor for lower counts of different leukocytes in very immature preterm infants. It is not clear if these low leukocyte counts are associated with a higher risk of neonatal infections or if lower numbers of inflammatory cells protect the lung and brain of very immature SGA infants by reducing inflammatory events postnatally.

  11. Trypanosoma cruzi infectivity assessment in "in vitro" culture systems by automated cell counting.

    PubMed

    Liempi, Ana; Castillo, Christian; Cerda, Mauricio; Droguett, Daniel; Duaso, Juan; Barahona, Katherine; Hernández, Ariane; Díaz-Luján, Cintia; Fretes, Ricardo; Härtel, Steffen; Kemmerling, Ulrike

    2015-03-01

    Chagas disease is an endemic, neglected tropical disease in Latin America that is caused by the protozoan parasite Trypanosoma cruzi. In vitro models constitute the first experimental approach to study the physiopathology of the disease and to assay potential new trypanocidal agents. Here, we report and describe clearly the use of commercial software (MATLAB(®)) to quantify T. cruzi amastigotes and infected mammalian cells (BeWo) and compared this analysis with the manual one. There was no statistically significant difference between the manual and the automatic quantification of the parasite; the two methods showed a correlation analysis r(2) value of 0.9159. The most significant advantage of the automatic quantification was the efficiency of the analysis. The drawback of this automated cell counting method was that some parasites were assigned to the wrong BeWo cell, however this data did not exceed 5% when adequate experimental conditions were chosen. We conclude that this quantification method constitutes an excellent tool for evaluating the parasite load in cells and therefore constitutes an easy and reliable ways to study parasite infectivity.

  12. Evaluation of mast cell counts and microvessel density in reactive lesions of the oral cavity

    PubMed Central

    Kouhsoltani, Maryam; Moradzadeh Khiavi, Monir; Tahamtan, Shabnam

    2016-01-01

    Background. Reliable immunohistochemical assays to assess the definitive role of mast cells (MCs) and angiogenesis in the pathogenesis of oral reactive lesions are generally not available. The aim of the present study was to evaluate mast cell counts (MCC) and microvessel density (MVD) in oral reactive lesions and determine the correlation between MCC and MVD. Methods. Seventy-five cases of reactive lesions of the oral cavity, including pyogenic granuloma, fibroma, peripheral giant cell granuloma, inflammatory fibrous hyperplasia, peripheral ossifying fibroma (15 for each category) were immunohisto-chemically stained with MC tryptase and CD31. Fifteen cases of normal gingival tissue were considered as the control group. The mean MCC and MVD in superficial and deep connective tissues were assessed and total MCC and MVD was computed for each lesion. Results. Statistically significant differences were observed in MCC and MVD between the study groups (P < 0.001). MC tryptase and CD31 expression increased in the superficial connective tissue of each lesion in comparison to the deep con-nective tissue. A significant negative correlation was not found between MCC and MVD in oral reactive lesions (P < 0.001, r = -0.458). Conclusion. Although MCs were present in the reactive lesions of the oral cavity, a direct correlation between MCC and MVD was not found in these lesions. Therefore, a significant interaction between MCs and endothelial cells and an active role for MCs in the growth of oral reactive lesions was not found in this study. PMID:28096950

  13. Basic Parameters of Blood Count as Prognostic Factors for Renal Cell Carcinoma

    PubMed Central

    Bryniarski, Piotr

    2016-01-01

    Background. Renal cell carcinoma is the most common type of kidney cancer. Taking account of morbidity and mortality increase, it is evident that searching for independent prognostic factors is needed. Aim of the Study. The aim of the study was to analyze routinely performed blood parameters as potential prognostic factors for kidney cancer. Material and Methods. We have retrospectively reviewed the records of 230 patients treated for renal cell carcinoma in the years 2000–2006. Preoperative blood parameters, postoperative histopathological results, and staging and grading were performed. To estimate the risk of tumor recurrence and cancer specific mortality (CSM) within five years of follow-up, uni- and multivariate Cox and regression analyses were used. To assess the quality of classifiers and to search for the optimal cut-off point, the ROC curve was used. Results. T stage of the tumor metastasis is the most important risk factor for early recurrence and cancer specific mortality (p < 0.001). The preoperative platelet count (PLT) above 351 × 103/uL (95.3%; 55.1%) and AUC of 77% are negative prognostic factors and correlate with increased cancer specific mortality (CSM) during the five-year follow-up (p < 0.001). Increased risk of local recurrence was observed for PLT above 243.5 × 103/ul (59%; 88%) and AUC of 80% (p = 0.001). The opposite was observed in the mean platelets volume (MPV) for cancer specific mortality (CSM). The cut-off point for the MPV was 10.1 fl (75.4%; 55.1%) and for the AUC is of 68.1% (p = 0.047). Conclusions. Many analyzed parameters in univariate regressions reached statistical significance and could be considered as potential prognostic factors for ccRCC. In multivariate analysis, only T stage, platelet count (PLT), and mean platelet volume (MPV) correlated with CSM or recurrent ccRCC. PMID:28105437

  14. Association between iron status and white blood cell counts in African schoolchildren of the North-West Province, South Africa.

    PubMed

    Onabanjo, Oluseye O; Jerling, Johann C; Covic, Namukolo; Van Graan, Averalda; Taljaard, Christine; Mamabolo, Ramoteme L

    2012-09-01

    Iron deficiency with or without anemia is associated with increased susceptibility to infection owing to impaired immune function; this study aimed to examine the associations between markers of iron status and white blood cell counts in African schoolchildren. This cross-sectional study is part of the larger BeForMi study done in the North-West province of South Africa. A total of 556 African schoolchildren (aged 7-10 years) were recruited from the three schools participating in the BeForMi multiple micronutrient intervention study. Demographic information of the children was obtained from their parents/caregivers/guardians in the language of choice using validated questionnaires. Anthropometric indices (weight and height), iron status parameters, hematological parameters (hemoglobin (Hb), red blood cell count (RBC), total and differential white blood cell counts) were measured using standard procedures. No significant gender differences were observed in most of the iron markers and hematological parameters except in C-reactive protein (CRP) (p=0.004) and eosinophils (p=0.042) which were higher in boys while RBC (p=0.018) and Hb (p=0.023) levels were higher in girls. No relationships were observed between the different iron markers and differential white blood cell counts. A positive correlation was observed between serum ferritin (SF) and CRP in girls only (r=0.336, p<0.01), and a positive correlation between SF and mean cell volume (MCV) in boys only (r=0.197, p<0.01). In both genders, no correlations were observed between the different iron markers and the differential white blood cell counts. The study revealed no associations between iron status and differential white blood cell counts in children that participated in the BeForMi study calling for more studies to be done in the area of the significance of iron supplementation in healthy children.

  15. Mechanical slowing-down of cytoplasmic diffusion allows in vivo counting of proteins in individual cells

    PubMed Central

    Okumus, Burak; Landgraf, Dirk; Lai, Ghee Chuan; Bakhsi, Somenath; Arias-Castro, Juan Carlos; Yildiz, Sadik; Huh, Dann; Fernandez-Lopez, Raul; Peterson, Celeste N.; Toprak, Erdal; El Karoui, Meriem; Paulsson, Johan

    2016-01-01

    Many key regulatory proteins in bacteria are present in too low numbers to be detected with conventional methods, which poses a particular challenge for single-cell analyses because such proteins can contribute greatly to phenotypic heterogeneity. Here we develop a microfluidics-based platform that enables single-molecule counting of low-abundance proteins by mechanically slowing-down their diffusion within the cytoplasm of live Escherichia coli (E. coli) cells. Our technique also allows for automated microscopy at high throughput with minimal perturbation to native physiology, as well as viable enrichment/retrieval. We illustrate the method by analysing the control of the master regulator of the E. coli stress response, RpoS, by its adapter protein, SprE (RssB). Quantification of SprE numbers shows that though SprE is necessary for RpoS degradation, it is expressed at levels as low as 3–4 molecules per average cell cycle, and fluctuations in SprE are approximately Poisson distributed during exponential phase with no sign of bursting. PMID:27189321

  16. Mechanical slowing-down of cytoplasmic diffusion allows in vivo counting of proteins in individual cells

    NASA Astrophysics Data System (ADS)

    Okumus, Burak; Landgraf, Dirk; Lai, Ghee Chuan; Bakhsi, Somenath; Arias-Castro, Juan Carlos; Yildiz, Sadik; Huh, Dann; Fernandez-Lopez, Raul; Peterson, Celeste N.; Toprak, Erdal; El Karoui, Meriem; Paulsson, Johan

    2016-05-01

    Many key regulatory proteins in bacteria are present in too low numbers to be detected with conventional methods, which poses a particular challenge for single-cell analyses because such proteins can contribute greatly to phenotypic heterogeneity. Here we develop a microfluidics-based platform that enables single-molecule counting of low-abundance proteins by mechanically slowing-down their diffusion within the cytoplasm of live Escherichia coli (E. coli) cells. Our technique also allows for automated microscopy at high throughput with minimal perturbation to native physiology, as well as viable enrichment/retrieval. We illustrate the method by analysing the control of the master regulator of the E. coli stress response, RpoS, by its adapter protein, SprE (RssB). Quantification of SprE numbers shows that though SprE is necessary for RpoS degradation, it is expressed at levels as low as 3-4 molecules per average cell cycle, and fluctuations in SprE are approximately Poisson distributed during exponential phase with no sign of bursting.

  17. Factors determining milk quality and implications for production structure under somatic cell count standard modification.

    PubMed

    Dong, F; Hennessy, D A; Jensen, H H

    2012-11-01

    Consumer and processor demand for high-quality milk has placed increasing pressure on US milk producers to achieve higher product standards. International standards for somatic cell count (SCC) are becoming more stringent, but in May 2011, the United States National Conference on Interstate Milk Shipments chose to retain the 750,000 cells/mL standard. Using ordinary least squares and quantile regressions on US Department of Agriculture Agricultural Resource Management Survey Dairy Costs and Returns Report data for 2005, we model producer and farm-level characteristics associated with SCC. Quantile regression analysis allows for a more parsed inquiry into statistical associations. Dairy Costs and Returns Report data provide cross-sectional information on the physical structure, input expenses, demographics, and outputs for farms in selected states. Location outside the Southeast, lower herd age, full-time farming status, use of biosecurity guidelines, good milking facilities and operations management, and application of related quality tests are all associated with lower SCC levels. Size of operation had little effect on SCC levels after controlling for other factors. Many of the operations that did not attain a more demanding SCC standard of 400,000 cells/mL had older operators, operators who expressed intention to exit within 10 yr, smaller size, and location in the Southeast when compared with those meeting the tighter standard. The results suggest that the stricter scheme favors larger farms that are more committed to production and are less likely to be sole or family proprietorships.

  18. Elevated white blood cell count and outcome in cancer patients with venous thromboembolism. Findings from the RIETE Registry.

    PubMed

    Trujillo-Santos, Javier; Di Micco, Pierpaolo; Iannuzzo, Mariateresa; Lecumberri, Ramón; Guijarro, Ricardo; Madridano, Olga; Monreal, Manuel

    2008-11-01

    A significant association between elevated white blood cell (WBC) count and mortality in patients with cancer has been reported, but the predictive value of elevated WBC on mortality in cancer patients with acute venous thromboembolism (VTE) has not been explored. RIETE is an ongoing registry of consecutive patients with acute VTE. We compared the three-month outcome of cancer patients with acute VTE according to their WBC count at baseline. As of May 2007, 3805 patients with active cancer and acute VTE had been enrolled in RIETE. Of them, 215 (5.7%) had low- (<4,000 cells/microl), 2,403 (63%) normal- (4,000-11,000 cells/microl), 1,187 (31%) elevated (>11,000 cells/microl) WBC count. During the study period 190 patients (5.0%) had recurrent VTE, 156 (4.1%) major bleeding, 889 (23%) died (399 of disseminated cancer, 113 of PE, 46 of bleeding. Patients with elevated WBC count at baseline had an increased incidence of recurrent VTE (odds ratio [OR]: 1.6; 95% confidence interval [CI]: 1.2-2.2), major bleeding (OR: 1.5; 95% CI: 1.1-2.1) or death (OR: 2.7; 95% CI: 2.3-3.2). Most of the reported causes of death were significantly more frequent in patients with elevated WBC count. Multivariate analysis confirmed that elevated WBC count was independently associated with an increased incidence of all three complications. In conclusion, cancer patients with acute VTE and elevated WBC count had an increased incidence of VTE recurrences, major bleeding or death. This worse outcome was consistent among all subgroups and persisted after multivariate adjustment.

  19. Mast Cell Hyperplasia and Eosinophilia Induced by Ascaris Body Fluid

    PubMed Central

    Archer, G. T.; Binet, J.-L.

    1971-01-01

    Daily i.p. injections of dilute Ascaris body fluid into rats induced peritoneal eosinophilia and the formation of pin-point follicles in the omentum. The follicles comprised plasma cells, macrophages and fibroblasts together with large numbers of eosinophils and mast cells. Electron microscopy of eosinophils in the follicles revealed loss of cytoplasmic granules and numerous vesicular and tubular structures in the cytoplasm. The mast cells showed clear areas round the granules, suggesting dissolution of granule components. ImagesFigs. 4-6Figs. 7-8Figs. 13-14Figs. 9-10Figs. 1-3Figs. 11-12 PMID:5135540

  20. Antiretroviral therapy suppressed participants with low CD4+ T-cell counts segregate according to opposite immunological phenotypes

    PubMed Central

    Pérez-Santiago, Josué; Ouchi, Dan; Urrea, Victor; Carrillo, Jorge; Cabrera, Cecilia; Villà-Freixa, Jordi; Puig, Jordi; Paredes, Roger; Negredo, Eugènia; Clotet, Bonaventura; Massanella, Marta; Blanco, Julià

    2016-01-01

    Background: The failure to increase CD4+ T-cell counts in some antiretroviral therapy suppressed participants (immunodiscordance) has been related to perturbed CD4+ T-cell homeostasis and impacts clinical evolution. Methods: We evaluated different definitions of immunodiscordance based on CD4+ T-cell counts (cutoff) or CD4+ T-cell increases from nadir value (ΔCD4) using supervised random forest classification of 74 immunological and clinical variables from 196 antiretroviral therapy suppressed individuals. Unsupervised clustering was performed using relevant variables identified in the supervised approach from 191 individuals. Results: Cutoff definition of CD4+ cell count 400 cells/μl performed better than any other definition in segregating immunoconcordant and immunodiscordant individuals (85% accuracy), using markers of activation, nadir and death of CD4+ T cells. Unsupervised clustering of relevant variables using this definition revealed large heterogeneity between immunodiscordant individuals and segregated participants into three distinct subgroups with distinct production, programmed cell-death protein-1 (PD-1) expression, activation and death of T cells. Surprisingly, a nonnegligible number of immunodiscordant participants (22%) showed high frequency of recent thymic emigrants and low CD4+ T-cell activation and death, very similar to immunoconcordant participants. Notably, human leukocyte antigen - antigen D related (HLA-DR) PD-1 and CD45RA expression in CD4+ T cells allowed reproducing subgroup segregation (81.4% accuracy). Despite sharp immunological differences, similar and persistently low CD4+ values were maintained in these participants over time. Conclusion: A cutoff value of CD4+ T-cell count 400 cells/μl classified better immunodiscordant and immunoconcordant individuals than any ΔCD4 classification. Immunodiscordance may present several, even opposite, immunological patterns that are identified by a simple immunological follow-up. Subgroup

  1. Comparison of cell counting methods in rodent pulmonary toxicity studies: automated and manual protocols and considerations for experimental design

    PubMed Central

    Zeidler-Erdely, Patti C.; Antonini, James M.; Meighan, Terence G.; Young, Shih-Houng; Eye, Tracy J.; Hammer, Mary Ann; Erdely, Aaron

    2016-01-01

    Pulmonary toxicity studies often use bronchoalveolar lavage (BAL) to investigate potential adverse lung responses to a particulate exposure. The BAL cellular fraction is counted, using automated (i.e. Coulter Counter®), flow cytometry or manual (i.e. hemocytometer) methods, to determine inflammatory cell influx. The goal of the study was to compare the different counting methods to determine which is optimal for examining BAL cell influx after exposure by inhalation or intratracheal instillation (ITI) to different particles with varying inherent pulmonary toxicities in both rat and mouse models. General findings indicate that total BAL cell counts using the automated and manual methods tended to agree after inhalation or ITI exposure to particle samples that are relatively nontoxic or at later time points after exposure to a pneumotoxic particle when the response resolves. However, when the initial lung inflammation and cytotoxicity was high after exposure to a pneumotoxic particle, significant differences were observed when comparing cell counts from the automated, flow cytometry and manual methods. When using total BAL cell count for differential calculations from the automated method, depending on the cell diameter size range cutoff, the data suggest that the number of lung polymorphonuclear leukocytes (PMN) varies. Importantly, the automated counts, regardless of the size cutoff, still indicated a greater number of total lung PMN when compared with the manual method, which agreed more closely with flow cytometry. The results suggest that either the manual method or flow cytometry would be better suited for BAL studies where cytotoxicity is an unknown variable. PMID:27251196

  2. 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.

  3. Genetic Modifiers of White Blood Cell Count, Albuminuria and Glomerular Filtration Rate in Children with Sickle Cell Anemia

    PubMed Central

    Flanagan, Jonathan M.; Alvarez, Ofelia A.; Nelson, Stephen C.; Aygun, Banu; Nottage, Kerri A.; George, Alex; Roberts, Carla W.; Piccone, Connie M.; Howard, Thad A.; Davis, Barry R.; Ware, Russell E.

    2016-01-01

    Discovery and validation of genetic variants that influence disease severity in children with sickle cell anemia (SCA) could lead to early identification of high-risk patients, better screening strategies, and intervention with targeted and preventive therapy. We hypothesized that newly identified genetic risk factors for the general African American population could also impact laboratory biomarkers known to contribute to the clinical disease expression of SCA, including variants influencing the white blood cell count and the development of albuminuria and abnormal glomerular filtration rate. We first investigated candidate genetic polymorphisms in well-characterized SCA pediatric cohorts from three prospective NHLBI-supported clinical trials: HUSTLE, SWiTCH, and TWiTCH. We also performed whole exome sequencing to identify novel genetic variants, using both a discovery and a validation cohort. Among candidate genes, DARC rs2814778 polymorphism regulating Duffy antigen expression had a clear influence with significantly increased WBC and neutrophil counts, but did not affect the maximum tolerated dose of hydroxyurea therapy. The APOL1 G1 polymorphism, an identified risk factor for non-diabetic renal disease, was associated with albuminuria. Whole exome sequencing discovered several novel variants that maintained significance in the validation cohorts, including ZFHX4 polymorphisms affecting both the leukocyte and neutrophil counts, as well as AGGF1, CYP4B1, CUBN, TOR2A, PKD1L2, and CD163 variants affecting the glomerular filtration rate. The identification of robust, reliable, and reproducible genetic markers for disease severity in SCA remains elusive, but new genetic variants provide avenues for further validation and investigation. PMID:27711207

  4. Human amniotic fluid stem cell preconditioning improves their regenerative potential.

    PubMed

    Rota, Cinzia; Imberti, Barbara; Pozzobon, Michela; Piccoli, Martina; De Coppi, Paolo; Atala, Anthony; Gagliardini, Elena; Xinaris, Christodoulos; Benedetti, Valentina; Fabricio, Aline S C; Squarcina, Elisa; Abbate, Mauro; Benigni, Ariela; Remuzzi, Giuseppe; Morigi, Marina

    2012-07-20

    Human amniotic fluid stem (hAFS) cells, a novel class of broadly multipotent stem cells that share characteristics of both embryonic and adult stem cells, have been regarded as promising candidate for cell therapy. Taking advantage by the well-established murine model of acute kidney injury (AKI), we studied the proregenerative effect of hAFS cells in immunodeficient mice injected with the nephrotoxic drug cisplatin. Infusion of hAFS cells in cisplatin mice improved renal function and limited tubular damage, although not to control level, and prolonged animal survival. Human AFS cells engrafted injured kidney predominantly in peritubular region without acquiring tubular epithelial markers. Human AFS cells exerted antiapoptotic effect, activated Akt, and stimulated proliferation of tubular cells possibly via local release of factors, including interleukin-6, vascular endothelial growth factor, and stromal cell-derived factor-1, which we documented in vitro to be produced by hAFS cells. The therapeutic potential of hAFS cells was enhanced by cell pretreatment with glial cell line-derived neurotrophic factor (GDNF), which markedly ameliorated renal function and tubular injury by increasing stem cell homing to the tubulointerstitial compartment. By in vitro studies, GDNF increased hAFS cell production of growth factors, motility, and expression of receptors involved in cell homing and survival. These findings indicate that hAFS cells can promote functional recovery and contribute to renal regeneration in AKI mice via local production of mitogenic and prosurvival factors. The effects of hAFS cells can be remarkably enhanced by GDNF preconditioning.

  5. PyMGC3: Finding stellar streams in the Galactic Halo using a family of Great Circle Cell counts methods

    NASA Astrophysics Data System (ADS)

    Mateu, C.

    2014-11-01

    PyMGC3 is a Python toolkit to apply the Modified Great Circle Cell Counts (mGC3) method to search for tidal streams in the Galactic Halo. The code computes pole count maps using the full mGC3/nGC3/GC3 family of methods. The original GC3 method (Johnston et al., 1996) uses positional information to search for 'great-circle-cell structures'; mGC3 makes use of full 6D data and nGC3 uses positional and proper motion data.

  6. 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

  7. Human Amniotic Fluid Stem Cell Preconditioning Improves Their Regenerative Potential

    PubMed Central

    Rota, Cinzia; Imberti, Barbara; Pozzobon, Michela; Piccoli, Martina; De Coppi, Paolo; Atala, Anthony; Gagliardini, Elena; Xinaris, Christodoulos; Benedetti, Valentina; Fabricio, Aline S.C.; Squarcina, Elisa; Abbate, Mauro; Benigni, Ariela; Remuzzi, Giuseppe

    2012-01-01

    Human amniotic fluid stem (hAFS) cells, a novel class of broadly multipotent stem cells that share characteristics of both embryonic and adult stem cells, have been regarded as promising candidate for cell therapy. Taking advantage by the well-established murine model of acute kidney injury (AKI), we studied the proregenerative effect of hAFS cells in immunodeficient mice injected with the nephrotoxic drug cisplatin. Infusion of hAFS cells in cisplatin mice improved renal function and limited tubular damage, although not to control level, and prolonged animal survival. Human AFS cells engrafted injured kidney predominantly in peritubular region without acquiring tubular epithelial markers. Human AFS cells exerted antiapoptotic effect, activated Akt, and stimulated proliferation of tubular cells possibly via local release of factors, including interleukin-6, vascular endothelial growth factor, and stromal cell–derived factor-1, which we documented in vitro to be produced by hAFS cells. The therapeutic potential of hAFS cells was enhanced by cell pretreatment with glial cell line–derived neurotrophic factor (GDNF), which markedly ameliorated renal function and tubular injury by increasing stem cell homing to the tubulointerstitial compartment. By in vitro studies, GDNF increased hAFS cell production of growth factors, motility, and expression of receptors involved in cell homing and survival. These findings indicate that hAFS cells can promote functional recovery and contribute to renal regeneration in AKI mice via local production of mitogenic and prosurvival factors. The effects of hAFS cells can be remarkably enhanced by GDNF preconditioning. PMID:22066606

  8. Time-course analysis of counts and degranulation of mast cells during early intestinal ischemia-reperfusion injury in mice.

    PubMed

    Ge, Mian; Gan, Xiaoliang; Liu, Dezhao; Zhang, Wenhua; Gao, Wanling; Huang, Pinjie; Hei, Ziqing

    2013-08-01

    Findings of previous studies have revealed that intestinal mucosal mast cells (IMMCs) are involved in small intestinal ischemia‑reperfusion injury (IIRI). However, time-course changes of mast cell counts and mast cell function in this process remain unclear. The present study aimed to observe the number of IMMCs and to investigate the correlation between their activation and small intestine injury at various time points during the period of small intestinal ischemia reperfusion (IIR). Healthy male Kunming mice were randomly divided into five groups, and were subjected to occlusion of the superior mesenteric artery (SMA) for 30 min and followed by reperfusion for 1, 3, 6 and 12 h. By contrast, the SMA was isolated but not clamped in the baseline group. Chiu's scores were assessed by light microscopy, tryptase protein and MCP7 protein expression in the intestine were quantified, and mast cell counts and levels of histamine and TNF-α in the intestine were measured. The results showed that IIR induced severe intestine injury within 12 h as demonstrated by Chiu's scores that was greatly increased as compared to the baseline group, accompanied by increased mast cell counts, histamine and TNF-α levels. However, the Chiu's scores were reduced in the IIR 12 h group compared with the IIR 1 h, IIR 3 h and IIR 6 h groups, with concomitant decreased mast cell counts, histamine and TNF-α levels. The tryptase and MCP7 protein expression was markedly increased in the IIR 1 h and IIR 3 h groups as compared with the baseline group, whereas this expression was gradually decreased at 6 and 12 h after reperfusion. The results of the present study suggest that IIR results in severe mucosal destruction within 6 h after reperfusion, associated with mast cell activation and substantial increases in the mast cell counts.

  9. Absolute lymphocyte count recovery after allogeneic hematopoietic stem cell transplantation predicts clinical outcome.

    PubMed

    Kim, Haesook T; Armand, Philippe; Frederick, David; Andler, Emily; Cutler, Corey; Koreth, John; Alyea, Edwin P; Antin, Joseph H; Soiffer, Robert J; Ritz, Jerome; Ho, Vincent T

    2015-05-01

    Immune reconstitution is critical for clinical outcome after allogeneic hematopoietic stem cell transplantation (HSCT). To determine the impact of absolute lymphocyte count (ALC) recovery on clinical outcomes, we conducted a retrospective study of 1109 adult patients who underwent a first allogeneic HSCT from 2003 through 2009, excluding patients who died or relapsed before day 30. The median age was 51 years (range, 18 to 74) with 52% undergoing reduced-intensity conditioning and 48% undergoing myeloablative conditioning HSCT with T cell-replete peripheral blood stem cells (93.7%) or marrow (6.4%) grafts. The median follow-up time was 6 years. To determine the threshold value of ALC for survival, the entire cohort was randomly split into a training set and a validation set in a 1:1 ratio, and then a restricted cubic spline smoothing method was applied to obtain relative hazard estimates of the relationship between ALC at 1 month and log hazard of progression-free survival (PFS). Based on this approach, ALC was categorized as ≤.2 × 10(9) cells/L (low) or >.2 × 10(9) cells/L. For patients with low ALC at 1, 2, or 3 months after HSCT, the overall survival (OS) (P ≤ .0001) and PFS (P ≤ .0002) were significantly lower and nonrelapse mortality (NRM) (P ≤ .002) was significantly higher compared with patients with ALC > .2 × 10(9) cells/L at each time point. When patients who had low ALC at 1, 2, or 3 months after HSCT were grouped together and compared, their outcomes were inferior to those of patients who had ALC > .2 × 10(9) cells/L at 1, 2, and 3 months after HSCT: the 5-year OS for patients with low ALC was 28% versus 46% for patients with ALC > .2 × 10(9) cells/L, P < .0001; the 5-year PFS was 21% versus 39%, P < .0001, respectively and 5-year NRM was 40% versus 18%, P < .0001, respectively. This result remained consistent when other prognostic factors, including occurrence of grade II to IV acute graft-versus-host disease (GVHD), were adjusted for in

  10. Prognostic impact of monocyte count at presentation in mantle cell lymphoma.

    PubMed

    von Hohenstaufen, Kathrin Aprile; Conconi, Annarita; de Campos, Cassio Polpo; Franceschetti, Silvia; Bertoni, Francesco; Margiotta Casaluci, Gloria; Stathis, Anastasios; Ghielmini, Michele; Stussi, Georg; Cavalli, Franco; Gaidano, Gianluca; Zucca, Emanuele

    2013-08-01

    An increased number of circulating monocytes at presentation has recently been associated with shorter survival in Hodgkin lymphoma, follicular lymphoma and diffuse large B cell lymphoma. This study aimed to assess the prognostic impact of the absolute monocyte count (AMC) at diagnosis in mantle cell lymphoma (MCL). AMC at diagnosis was available in 97 MCL cases recorded in the databases of the Oncology Institute of Southern Switzerland in Bellinzona (Switzerland) and the Division of Haematology of the Amedeo Avogadro University of Eastern Piedmont in Novara (Italy). With a median follow up of 7 years, the 5-year overall survival was 29% for patients with AMC >0·50 × 10(9) /l and 62% for patients with AMC ≤0·50 × 10(9) /l (P = 0·008). Elevated AMC and beta-2 microglobulin at diagnosis remained independent outcome predictors at multivariate analysis, controlling for the MCL International Prognostic Index (MIPI), and have been used to build a simple prognostic scoring system. In this relatively small and heterogeneous series an increased AMC identified poor-risk patients. Our results suggest that AMC together with the beta-2 microglobulin level might provide an inexpensive way to stratify MCL patient risk as a complement to the MIPI, which was confirmed to be a very powerful prognostic tool.

  11. Effects of diabetic keratopathy on corneal optical density, central corneal thickness, and corneal endothelial cell counts.

    PubMed

    Gao, Feng; Lin, Tao; Pan, Yingzhe

    2016-09-01

    Diabetic keratopathy is an ocular complication that occurs with diabetes. In the present study, the effect of diabetic keratopathy on corneal optical density, central corneal thickness, and corneal endothelial cell count was investigated. One hundred and eighty diabetic patients (360 eyes) were enrolled in the study during the period from March, 2012 to March, 2013. The patients were divided into three age groups: <5, 5-10 and >10 years, with 60 patients per group (120 eyes). During the same period, 60 healthy cases (120 eyes) were selected and labeled as the normal control group. The Pentacam was used to measure the corneal optical density, and central corneal thickness. Specular microscopy was used to examine the corneal endothelial cell density. The coefficient of partial correlation was used to control age and correlate the analysis between the corneal optical density, corneal endothelial cell density, and central corneal thickness. The stage of the disease, the medial and intimal corneal optical density and central corneal thickness was analyzed in the diabetes group. The corneal optical density in the diabetes group increased compared with that of the normal control group. The medial and intimal corneal optical density and central corneal thickness were positively correlated with the course of the disease. However, the corneal endothelial cell density was not associated with the course of diabetes. There was a positive association between the medial and intimal corneal optical density and central corneal thickness of the diabetic patients. In conclusion, the results of the present study show that medial and intimal corneal optical density and central corneal thickness were sensitive indicators for early diabetic keratopathy.

  12. Effects of diabetic keratopathy on corneal optical density, central corneal thickness, and corneal endothelial cell counts

    PubMed Central

    Gao, Feng; Lin, Tao; Pan, Yingzhe

    2016-01-01

    Diabetic keratopathy is an ocular complication that occurs with diabetes. In the present study, the effect of diabetic keratopathy on corneal optical density, central corneal thickness, and corneal endothelial cell count was investigated. One hundred and eighty diabetic patients (360 eyes) were enrolled in the study during the period from March, 2012 to March, 2013. The patients were divided into three age groups: <5, 5–10 and >10 years, with 60 patients per group (120 eyes). During the same period, 60 healthy cases (120 eyes) were selected and labeled as the normal control group. The Pentacam was used to measure the corneal optical density, and central corneal thickness. Specular microscopy was used to examine the corneal endothelial cell density. The coefficient of partial correlation was used to control age and correlate the analysis between the corneal optical density, corneal endothelial cell density, and central corneal thickness. The stage of the disease, the medial and intimal corneal optical density and central corneal thickness was analyzed in the diabetes group. The corneal optical density in the diabetes group increased compared with that of the normal control group. The medial and intimal corneal optical density and central corneal thickness were positively correlated with the course of the disease. However, the corneal endothelial cell density was not associated with the course of diabetes. There was a positive association between the medial and intimal corneal optical density and central corneal thickness of the diabetic patients. In conclusion, the results of the present study show that medial and intimal corneal optical density and central corneal thickness were sensitive indicators for early diabetic keratopathy. PMID:27588090

  13. Choroid plexus carcinoma cells in the cerebrospinal fluid of a Staffordshire Bull Terrier.

    PubMed

    Pastorello, Alice; Constantino-Casas, Fernando; Archer, Joy

    2010-12-01

    An 11-year-old female intact Staffordshire Bull Terrier was referred to the Queen's Veterinary School Hospital at the University of Cambridge with sudden onset of episodic behavioral changes, a mammary mass, and papilledema in the right eye. On physical examination the dog appeared depressed and had a head tilt to the right with anisocoria. Using magnetic resonance imaging, a broad-based lesion that obliterated the fourth ventricle was detected in the right brainstem. There was no evidence of pulmonary metastasis. Cerebrospinal fluid (CSF) was then obtained; fluid analysis showed an increased cell count (165 cells/μL, reference interval 0-7 cells/μL) and total protein (0.30 g/L, reference value <0.25 g/L). Cytologic evaluation revealed a population of atypical epithelial cells arranged in cohesive rafts and characterized by moderate to occasionally marked anisocytosis and anisokaryosis. The appearance was highly suspicious of a malignant epithelial neoplasm. The dog was euthanized and on postmortem examination an asymmetrical nonencapsulated cerebellar mass was found within the choroid plexus of the fourth ventricle with local extension into the cerebellopontine angle. Histologic sections of the cerebellar mass contained arborizing papillary structures covered by a single layer of atypical epithelial cells that showed local infiltration into the adjacent neuropil. The diagnosis was choroid plexus carcinoma. The atypical epithelial cells were negative for pancytokeratin and strongly positive for vimentin. The finding of clusters of choroid plexus epithelial cells in the CSF demonstrates the value of utilizing a relatively noninvasive diagnostic technique for diagnosis of choroid plexus tumors.

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

    SciTech Connect

    Aoki, Shigehisa; Ikeda, Satoshi; Takezawa, Toshiaki; Kishi, Tomoya; Makino, Junichi; Uchihashi, Kazuyoshi; Matsunobu, Aki; Noguchi, Mitsuru; Sugihara, Hajime; Toda, Shuji

    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

  15. Evaluation of Tumor Cell Proliferation by Ki-67 Expression and Mitotic Count in Lymph Node Metastases from Breast Cancer

    PubMed Central

    Aziz, Sura; Wik, Elisabeth; Davidsen, Benedicte; Aas, Hans; Aas, Turid; Akslen, Lars A.

    2016-01-01

    Few studies have addressed the risk of recurrence by assessing proliferation markers in lymph node metastasis from breast cancer. Here, we aimed to examine Ki-67 expression and mitotic count in lymph nodes in comparison with primary tumors. A cohort of node positive breast cancer (n = 168) was studied as a part of the prospective Norwegian Breast Cancer Screening Program (1996–2009). The percentage of Ki-67 positivity was counted per 500 tumor cells in hot-spot areas (x630). Mitotic count was conducted in the most cellular and mitotic active areas in 10 high power fields (x400). Our results showed that Ki-67 and mitotic count were significantly correlated between primary tumor and lymph nodes (Spearman`s correlation 0. 56 and 0.46, respectively) and were associated with most of the histologic features of the primary tumor. Univariate survival analysis (log-rank test) showed that high Ki-67 and mitotic count in the primary tumor and lymph node metastasis significantly predicted risk of recurrence. In multivariate analysis, mitotic count in the lymph node metastasis was an independent predictor of tumor recurrence. In conclusion, proliferation markers in lymph node metastases significantly predicted disease free survival in node positive breast cancer. PMID:26954367

  16. Evaluation of Tumor Cell Proliferation by Ki-67 Expression and Mitotic Count in Lymph Node Metastases from Breast Cancer.

    PubMed

    Aziz, Sura; Wik, Elisabeth; Knutsvik, Gøril; Klingen, Tor Audun; Chen, Ying; Davidsen, Benedicte; Aas, Hans; Aas, Turid; Akslen, Lars A

    2016-01-01

    Few studies have addressed the risk of recurrence by assessing proliferation markers in lymph node metastasis from breast cancer. Here, we aimed to examine Ki-67 expression and mitotic count in lymph nodes in comparison with primary tumors. A cohort of node positive breast cancer (n = 168) was studied as a part of the prospective Norwegian Breast Cancer Screening Program (1996-2009). The percentage of Ki-67 positivity was counted per 500 tumor cells in hot-spot areas (x630). Mitotic count was conducted in the most cellular and mitotic active areas in 10 high power fields (x400). Our results showed that Ki-67 and mitotic count were significantly correlated between primary tumor and lymph nodes (Spearman`s correlation 0. 56 and 0.46, respectively) and were associated with most of the histologic features of the primary tumor. Univariate survival analysis (log-rank test) showed that high Ki-67 and mitotic count in the primary tumor and lymph node metastasis significantly predicted risk of recurrence. In multivariate analysis, mitotic count in the lymph node metastasis was an independent predictor of tumor recurrence. In conclusion, proliferation markers in lymph node metastases significantly predicted disease free survival in node positive breast cancer.

  17. 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.

  18. Evaluation of volume and total nucleated cell count as cord blood selection parameters: a receiver operating characteristic curve modeling approach.

    PubMed

    Jaime-Pérez, José C; Monreal-Robles, Roberto; Rodríguez-Romo, Laura N; Mancías-Guerra, Consuelo; Herrera-Garza, José Luís; Gómez-Almaguer, David

    2011-11-01

    The objective of the study was to evaluate the current standard practice of using volume and total nucleated cell (TNC) count for the selection of cord blood (CB) units for cryopreservation and further transplantation. Data on 794 CB units whose CD34+ cell content was determined by flow cytometry were analyzed by using a receiver operating characteristic (ROC) curve model to validate the performance of volume and TNC count for the selection of CB units with grafting purposes. The TNC count was the best parameter to identify CB units having 2 × 10(6) or more CD34+ cells, with an area under the ROC curve of 0.828 (95% confidence interval, 0.800-0.856; P < .01) and an efficiency of 75.4%. Combination of parameters (TNC/mononuclear cells [MNCs], efficiency 74.7%; TNC/volume, efficiency 68.9%; and volume/MNCs, efficiency 68.3%) did not lead to improvement in CB selection. All CB units having a TNC count of 8 × 10(8) or more had the required CD34+ cell dose for patients weighing 10 kg or less.

  19. Hematologic variables and venous thrombosis: red cell distribution width and blood monocyte count are associated with an increased risk.

    PubMed

    Rezende, Suely Meireles; Lijfering, Willem M; Rosendaal, Frits R; Cannegieter, Suzanne C

    2014-01-01

    Recent studies suggest that leukocytes and erythrocytes play a role in coagulation. However, whether leukocytes, erythrocytes and other hematologic variables are associated with risk of venous thrombosis is not well known. To study this, we used data from 2473 patients with venous thrombosis and 2935 controls. The variables assessed were: total leukocytes, granulocytes, lymphocytes, monocytes, hematocrit, hemoglobin, erythrocytes and red cell indices (mean corpuscular volume, mean corpuscular hemoglobin, mean corpuscular hemoglobin concentration and red cell distribution width). We found a strong dose-response relation for higher red cell distribution width and monocyte count with risk of venous thrombosis, with odds ratios of 3.1 (95% confidence interval, 2.0-4.8) and 2.8 (95% confidence interval, 1.3-5.8), respectively, after adjustment for age, sex, C-reactive protein level, malignancy and co-morbidities. Monocyte count and red cell distribution width were associated with venous thrombosis even within reference ranges. A low monocyte count (<0.12 × 10(9)/L) was associated with a lower risk of venous thrombosis after full adjustment (odds ratios 0.6; 95% confidence interval, 0.4-0.8). In summary, high red cell distribution width and blood monocyte count, two parameters that are inexpensive and easily obtainable, were clearly associated with an increased risk of venous thrombosis. Future studies should evaluate the underlying mechanism and the use of these variables in prediction models for first and recurrent thrombosis.

  20. [Experiences with a sedimentation technique for the enrichment of cerebrospinal fluid cells in the dog and cat. Part 1].

    PubMed

    Grevel, V

    1991-10-01

    The differentiation of cells of the cerebrospinal fluid (CSF) gives valuable information about certain neurologic diseases. The sedimentation technique of Sayk modified by Kölmel is introduced and its application described. Cells of 300 samples of CSF from dogs and cats are evaluated. There were very good to reasonable results in 88% (263 of 300) of the samples. A comparison between cell number, morphology and protein content in 150 samples was performed. 90 CSF samples with normal cell count (less than or equal to 5/mm3, determined in the Fuchs-Rosenthal chamber) were compared with the cell yield after sedimentation. In 58% (52 of 90) of the samples more than 200 cells were found, in 13% (12 of 90) more than 800 could be differentiated. The results are compared with those of other methods mentioned in the literature.

  1. Multilineage potential research of bovine amniotic fluid mesenchymal stem cells.

    PubMed

    Gao, Yuhua; Zhu, Zhiqiang; Zhao, Yuhua; Hua, Jinlian; Ma, Yuehui; Guan, Weijun

    2014-02-28

    The use of amnion and amniotic fluid (AF) are abundant sources of mesenchymal stem cells (MSCs) that can be harvested at low cost and do not pose ethical conflicts. In human and veterinary research, stem cells derived from these tissues are promising candidates for disease treatment, specifically for their plasticity, their reduced immunogenicity, and high anti-inflammatory potential. This work aimed to obtain and characterize bovine amniotic fluid mesenchymal stem cells (AFMSC). The bovine AF from the amniotic cavity of pregnant gilts in the early stages of gestation (3- and 4-m-old bovine embryos) was collected. AFMSCs exhibit a fibroblastic-like morphology only starting from the fourth passage, being heterogeneous during the primary culture. Immunofluorescence results showed that AFMSCs were positive for β-integrin, CD44, CD73 and CD166, but negative for CD34, CD45. Meanwhile, AFMSCs expressed ES cell markers, such as Oct4, and when appropriately induced, are capable of differentiating into ectodermal and mesodermal lineages. This study reinforces the emerging importance of these cells as ideal tools in veterinary medicine; future studies aimed at a deeper evaluation of their immunological properties will allow a better understanding of their role in cellular therapy.

  2. A robust cell counting approach based on a normalized 2D cross-correlation scheme for in-line holographic images.

    PubMed

    Ra, Ho-Kyeong; Kim, Hyungseok; Yoon, Hee Jung; Son, Sang Hyuk; Park, Taejoon; Moon, Sangjun

    2013-09-07

    To achieve the important aims of identifying and marking disease progression, cell counting is crucial for various biological and medical procedures, especially in a Point-Of-Care (POC) setting. In contrast to the conventional manual method of counting cells, a software-based approach provides improved reliability, faster speeds, and greater ease of use. We present a novel software-based approach to count in-line holographic cell images using the calculation of a normalized 2D cross-correlation. This enables fast, computationally-efficient pattern matching between a set of cell library images and the test image. Our evaluation results show that the proposed system is capable of quickly counting cells whilst reliably and accurately following human counting capability. Our novel approach is 5760 times faster than manual counting and provides at least 68% improved accuracy compared to other image processing algorithms.

  3. [Isolation and gene modification of amniotic fluid derived progenitor cells].

    PubMed

    Yang, Chenmin; Fan, Shuyue; Tang, Huixiang; Gong, Zhijuan; Gong, Xiuli; Ren, Zhaorui; Zeng, Fanyi

    2014-03-01

    We established methods to isolate human amniotic fluid-derived progenitor cells (hAFPCs), and analyze the ability of hAFPCs to secrete human coagulation factor IX (hFIX) after gene modification. The hAFPCs were manually isolated by selection for attachment to gelatin coated culture dish. hFIX cDNA was transfected into hAPFCs by using a lentiviral vector. The hFIX protein concentration and activity produced from hAFPCs were determined by enzyme-linked immunosorbent assay (ELISA) and clotting assay. The isolated spindle-shaped cells showed fibroblastoid morphology after three culture passages. The doubling time in culture was 39.05 hours. Immunocytochemistry staining of the fibroblast-like cells from amniotic fluid detected expression of stem cell markers such as SSEA4 and TRA1-60. Quantitative PCR analysis demonstrated the expression of NANOG, OCT4 and SOX2 mRNAs. Transfected hAFPCs could produce and secrete hFIX into the culture medium. The observed concentration of secreted hFIX was 20.37% +/- 2.77% two days after passage, with clotting activity of 16.42% +/- 1.78%. The amount of hFIX:Ag reached a plateau of 50.35% +/- 5.42%, with clotting activity 45.34% +/- 4.67%. In conclusion, this study established method to isolate and culture amniotic fluid progenitor cells. Transfected hAFPCs can produce hFIX at stable levels in vitro, and clotting activity increases with higher hFIX concentration. Genetically engineered hAFPC are a potential method for prenatal treatment of hemophilia B.

  4. 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) ...

  5. Mesenchymal Stem Cells from Wharton's Jelly and Amniotic Fluid.

    PubMed

    Joerger-Messerli, Marianne S; Marx, Caterina; Oppliger, Byron; Mueller, Martin; Surbek, Daniel V; Schoeberlein, Andreina

    2016-02-01

    The discovery of mesenchymal stem cells (MSCs) in perinatal sources, such as the amniotic fluid (AF) and the umbilical connective tissue, the so-called Wharton's jelly (WJ), has transformed them into promising stem cell grafts for the application in regenerative medicine. The advantages of AF-MSCs and WJ-MSCs over adult MSCs, such as bone marrow-derived mesenchymal stem cells (BM-MSCs), include their minimally invasive isolation procedure, their more primitive cell character without being tumourigenic, their low immunogenicity and their potential autologous application in congenital disorders and when cryopreserved in adulthood. This chapter gives an overview of the biology of AF-MSCs and WJ-MSCs, and their regenerative potential based on the results of recent preclinical and clinical studies. In the end, open questions concerning the use of WJ-MSCs and AF-MSCs in regenerative medicine will be emphasized.

  6. Association between BoLA-DRB3 and somatic cell count in Holstein cattle from Argentina.

    PubMed

    Baltian, L R; Ripoli, M V; Sanfilippo, S; Takeshima, S N; Aida, Y; Giovambattista, G

    2012-07-01

    Different studies have proved that the resistance/susceptibility to mastitis is genetically determined. The major histocompatibility complex in cows is known as bovine lymphocyte antigen (BoLA). Genes from the BoLA have been associated with the occurrence of infectious diseases such as mastitis and leukosis, especially the BoLA-DRB gene. The object of the present study was to detect associations between BoLA-DRB3 alleles and somatic cell count (SCC), as an indicator of resistance/susceptibility to mastitis in Holstein cattle (N = 123) from La Pampa, Argentina. Fisher's exact test and Woolf-Haldane odds ratio were applied to study the association between SCC and BoLA-DRB3 allele frequencies. Significant association was noted between BoLA-DRB3.2*23 and *27 alleles (p < 0.05) and protective or susceptibility effects, respectively. In addition, alleles BoLA-DRB3.2*20 and *25 exhibit suggestive association with high SCC (p < 0.1). These results were partially in agreement with data reported from Japanese Holstein cattle, but differed from those published by other authors. A possible explanation for the contrasting results could be that the mastitis is a multifactor disease caused by different pathogens. Moreover, most of the studies were carried out using PCR-RFLP method, which has less resolution than PCR-SBT because PCR-RFLP defined alleles included more than one sequenced alleles.

  7. Preoperative Aspartate Aminotransferase to White Blood Cell Count Ratio Predicting Postoperative Outcomes of Hepatocellular Carcinoma.

    PubMed

    Liao, Weijia; Wang, Yongqin; Liao, Yan; He, Songqing; Jin, Junfei

    2016-04-01

    Effective biomarkers for predicting prognosis of hepatocellular carcinoma (HCC) patients after hepatectomy is urgently needed. The purpose of this study is to evaluate the value of the preoperative peripheral aspartate aminotransferase to white blood cell count ratio (AWR) for the prognostication of patients with HCC.Clinical data of 396 HCC patients who underwent radical hepatectomy were retrospectively analyzed. The patients were divided into the low-AWR group (AWR ≤5.2) and the high-AWR group (AWR >5.2); univariate analysis, Kaplan-Meier method analysis, and the multivariate analysis by Cox regression were conducted, respectively.The results showed that AWR was associated with alpha-fetoprotein (AFP), tumor size, Barcelona clinic liver cancer (BCLC) stage, portal vein tumor thrombus (PVTT), and alanine aminotransferase (ALT) in HCC. AWR > 5.2, AFP > 100 ng/mL, size of tumor >6 cm, number of multiple tumors, B-C of BCLC stage, PVTT, and distant metastasis were predictors of poorer disease-free survival (DFS) and overall survival (OS). Except for recurrence, which was an independent predictor for OS only, AWR >5.2, size of tumor >6 cm, and PVTT were independent predictors of both DFS and OS.We concluded that preoperative AWR > 5.2 was an adverse predictor of DFS and OS in HCC after hepatectomy, AWR might be a novel prognostic biomarker in HCC after curative resection.

  8. Vascular mechanobiology: endothelial cell responses to fluid shear stress.

    PubMed

    Ando, Joji; Yamamoto, Kimiko

    2009-11-01

    Endothelial cells (ECs) lining blood vessel walls respond to shear stress, a fluid mechanical force generated by flowing blood, and the EC responses play an important role in the homeostasis of the circulatory system. Abnormal EC responses to shear stress impair various vascular functions and lead to vascular diseases, including hypertension, thrombosis, and atherosclerosis. Bioengineering approaches in which cultured ECs are subjected to shear stress in fluid-dynamically designed flow-loading devices have been widely used to analyze EC responses at the cellular and molecular levels. Remarkable progress has been made, and the results have shown that ECs alter their morphology, function, and gene expression in response to shear stress. Shear stress affects immature cells, as well as mature ECs, and promotes differentiation of bone-marrow-derived endothelial progenitor cells and embryonic stem cells into ECs. Much research has been done on shear stress sensing and signal transduction, and their molecular mechanisms are gradually coming to be understood. However, much remains uncertain, and many candidates have been proposed for shear stress sensors. More extensive studies of vascular mechanobiology should increase our understanding of the molecular basis of the blood-flow-mediated control of vascular functions.

  9. Fluid Flow Induced Calcium Response in Bone Cell Network

    PubMed Central

    Huo, Bo; Lu, Xin L.; Hung, Clark T.; Costa, Kevin D.; Xu, Qiaobing; Whitesides, George M.; Guo, X. Edward

    2010-01-01

    In our previous work, bone cell networks with controlled spacing and functional intercellular gap junctions had been successfully established by using microcontact printing and self assembled monolayers technologies [Guo, X. E., E. Takai, X. Jiang, Q. Xu, G. M. Whitesides, J. T. Yardley, C. T. Hung, E. M. Chow, T. Hantschel, and K. D. Costa. Mol. Cell. Biomech. 3:95–107, 2006]. The present study investigated the calcium response and the underlying signaling pathways in patterned bone cell networks exposed to a steady fluid flow. The glass slides with cell networks were separated into eight groups for treatment with specific pharmacological agents that inhibit pathways significant in bone cell calcium signaling. The calcium transients of the network were recorded and quantitatively evaluated with a set of network parameters. The results showed that 18α-GA (gap junction blocker), suramin (ATP inhibitor), and thapsigargin (depleting intracellular calcium stores) significantly reduced the occurrence of multiple calcium peaks, which were visually obvious in the untreated group. The number of responsive peaks also decreased slightly yet significantly when either the COX-2/PGE2 or the NOS/nitric oxide pathway was disrupted. Different from all other groups, cells treated with 18α-GA maintained a high concentration of intracellular calcium following the first peak. In the absence of calcium in the culture medium, the intracellular calcium concentration decreased slowly with fluid flow without any calcium transients observed. These findings have identified important factors in the flow mediated calcium signaling of bone cells within a patterned network. PMID:20852730

  10. 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.

  11. Red blood cell count has an independent contribution to the prediction of ultrasonography-diagnosed fatty liver disease

    PubMed Central

    Wu, Shang-ling; Liao, Gong-cheng; Fang, Ai-ping; Zhu, Ming-fan; Zhu, Hui-lian

    2017-01-01

    Background & aims Red blood cell (RBC) indices have been demonstrated to be associated with fatty liver disease (FLD) and metabolic syndrome. However, controversy exists regarding the relationship of RBC indices with FLD to date and few has focused on RBC count. This study aimed to explore the association between RBC count and risk of FLD in Southern Chinese adults. Methods A hospital-based cross-sectional study was performed in two hospital health examination centers, including information on ultrasonography-diagnosed FLD, anthropometric indices and biochemical measurements. Covariance analysis was used to evaluate group differences. After quintile classification of RBC counts, logistic regression analysis was conducted to evaluate the odds ratios (ORs) of FLD. Results This study consisted of 8618 subjects (4137 men and 4481 women) aged between 20 and 89 years. FLD cases had higher RBC counts than non-FLD cases in both genders (P<0.001). The prevalence rates of FLD increased with the RBC quintiles in both genders (all P trend<0.001), and were higher in men than women. Binary logistic regression analysis showed positive association between RBC count and FLD, and the OR (95% confidence interval (CI)) were 2.56 (2.06–3.18) in men and 3.69 (2.74–4.98) in women, respectively, when comparing Q5 with Q1. Stratified analyses showed similar trends among subjects with and without FLD risk factors. Gender independent results were similar to gender dependent results. Conclusions Elevated RBC count is independently associated with high risk of FLD, suggesting that the RBC count may be a potential risk predictor for FLD. PMID:28187211

  12. 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 297,000 cells/mL in 2012 and 274,000 cells/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

  13. Innate immunity in free-ranging African buffalo (Syncerus caffer): associations with parasite infection and white blood cell counts.

    PubMed

    Beechler, Brianna R; Broughton, Heather; Bell, Austin; Ezenwa, Vanessa O; Jolles, Anna E

    2012-01-01

    Mammalian immunology has been studied in great detail in laboratory animals, but few of the tools and less of the insight derived from these studies have been placed in the context of natural, outbred wildlife populations subject to variable environments. We investigated patterns of innate immunity in free-ranging African buffalo in relation to host traits (age, reproductive status, body condition, white blood cell counts) and disease status (bovine tuberculosis [BTB], gastrointestinal nematodes, coccidia, ticks). We evaluated and used an in vitro assay measuring bactericidal competence of blood to assess a component of innate immunity in 200 female buffalo captured at Kruger National Park, South Africa, in June/July and October 2008. Animals with BTB had higher bactericidal competence of blood. Animals with higher neutrophil counts had higher bactericidal competence, whereas animals with lower lymphocyte counts had higher bactericidal competence. This pattern was driven by animals captured at the end of the dry season (October) and may be evidence of immune polarization, whereby individuals are unable to upregulate multiple components of immunity simultaneously. Bactericidal competence did not vary with host pregnancy status, body condition, age, lactation, tick infestation, nematode egg count, or coccidia oocyst count. Overall, we demonstrate that the bactericidal competence assay is practical and informative for field-based studies in wild bovids. Our results also show a correlation between bactericidal competence and bovine tuberculosis infection and reveal possible functional polarizations between different types of immune response in a free-ranging mammal.

  14. The severity, extent and recurrence of necrotizing periodontal disease in relation to HIV status and CD4+ T cell count.

    PubMed

    Phiri, Reality; Feller, Liviu; Blignaut, Elaine

    2010-10-01

    South Africa ranks among the three countries with the highest prevalence of HIV infection in sub-Saharan Africa, with an estimated 29.5% of women attending antenatal clinics being infected. Necrotizing periodontal disease is a well recognized HIV-associated oral condition. The objective of this investigation was to determine a possible correlation between the extent, severity and treatment outcome of necrotizing periodontal disease in relation to a person's HIV status and CD4+ T cell count. Data from 105 consecutive patients presenting with necrotizing periodontal disease at an academic oral health centre in South Africa were analysed. All patients were provided with an opportunity to undergo voluntary counseling and testing for HIV infection, were treated for necrotizing periodontal disease and followed over a period of nine months. The mean age of the cohort was 28 years old (range 12 - 52). Of 98 (93.3%) patients unaware of their HIV serostatus at the initial visit, 59 (56.2%) consented to testing. In total 45 (42.9%) were HIV-seropositive with a mean CD4+ T cell count of 222.7 cells/microl and 14 (13.3%) were HIV-seronegative, with a significantly higher mean CD4+ T cell count of 830 cells/microl (Fisher's exact test, p < 0.001), while the status of 46 (43.8%) remained unknown. In 101 (96.2%) patients, > or = 5 tooth sites were affected, and in 27 (26%) > or = 4 mm of gingival tissue were affected. This study, which included HIV-seropositive, HIV-seronegative and persons of unknown HIV status, revealed no statistical evidence that HIV infection was associated with the extent, severity or relapse of necrotizing periodontal disease. No statistically significant association could be demonstrated between the extent, severity and recurrence of necrotizing periodontal disease and a CD4+ T cell count < or = 200 cells/microl among HIV-seropositive patients.

  15. Relationship between somatic cell count and milk yield in different stages of lactation.

    PubMed

    Hagnestam-Nielsen, C; Emanuelson, U; Berglund, B; Strandberg, E

    2009-07-01

    The association between somatic cell count (SCC) and daily milk yield in different stages of lactation was investigated in cows free of clinical mastitis (CM). Data were recorded between 1989 and 2004 in a research herd, and consisted of weekly test-day (TD) records from 1,155 lactations of Swedish Holstein and Swedish Red cows. The main data set (data set A) containing 36,117 records excluded TD affected by CM. In this data set, the geometric mean SCC was 55,000 and 95,000 cells/mL in primiparous and multiparous cows, respectively. A subset of data set A (data set B), containing 27,753 records excluding all TD sampled in lactations affected by CM, was created to investigate the effect of subclinical mastitis (SCM) in lactations free of CM. Daily milk yields were analyzed using a mixed linear model with lactation stage; linear, quadratic and cubic regressions of log(2)-transformed and centered SCC nested within lactation stage; weeks in lactation; TD season; parity; breed; pregnancy status; year-season of calving; calving, reproductive, metabolic and claw disorders; and housing system as fixed effects. A random regression was included to further improve the modeling of the lactation curve. Primiparous and multiparous cows were analyzed separately. The magnitude of daily milk loss associated with increased SCC depended on stage of lactation and parity, and was most extensive in late lactation irrespective of parity. In data set A, daily milk loss at an SCC of 500,000 cells/mL ranged from 0.7 to 2.0 kg (3 to 9%) in primiparous cows, depending on stage of lactation. In multiparous cows, corresponding loss was 1.1 to 3.7 kg (4 to 18%). Regression coefficients of primiparous cows estimated from data set B were consistent with those obtained from data set A, whereas data set B generated more negative regression coefficients of multiparous cows suggesting a higher milk loss associated with increased SCC in lactations in which the cow did not develop CM. The 305-d milk

  16. 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, β-hydroxybutyrate sensor, and other sensor systems). The AMS farms had a higher average SCC (by 12,000 cells/mL) after sensor investment, and CMS farms with a mastitis detection system had a lower average SCC (by 10,000 cells/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

  17. Confocal Raman microscopy of pathologic cells in cerebrospinal fluid

    NASA Astrophysics Data System (ADS)

    Gonchukov, S. A.; Lonkina, T. V.; Minaeva, S. A.; Sundukov, A. V.; Migmanov, T. E.; Lademann, J.; Darvin, M. E.; Bagratashvili, V. N.

    2014-01-01

    In this work, the spatial localization of leucocytes, bacteria, and erythrocytes in the crystal pattern of a dried droplet of cerebrospinal fluid (CSF) is established. Characteristic lines are detected and identified in the Raman spectrum of the CSF that point to the presence of pathologic cells therein and can be used in a timely way to diagnose meningitis, the spectroscopic sample preparation procedure being simple enough. A dry CSF sample retains its characteristic spectral features for no less than three days, which is important for its safe keeping and transportation, and also for the computer processing of its spectra.

  18. Blood Count Tests

    MedlinePlus

    ... white blood cells (WBC), and platelets. Blood count tests measure the number and types of cells in ... helps doctors check on your overall health. The tests can also help to diagnose diseases and conditions ...

  19. Relationship between white blood cell count and components of metabolic syndrome among young adolescents.

    PubMed

    Wu, Chung-Ze; Hsiao, Fone-Ching; Lin, Juinn-Diann; Su, Ching-Chieh; Wang, Kwo-Syin; Chu, Yi-Min; Lee, Li-Hsiu; Wang, Kun; Hsia, Te-Lin; Pei, Dee

    2010-03-01

    Components of metabolic syndrome (MetS) have been associated with several inflammatory factors, including white blood cell count (WBCC). In the present study, the relationships between WBCC and aspects of MetS in young adolescents were investigated. We enrolled 596 participants (328 males and 268 females) from 10 to 13 years of age and with normal WBCC in this study. They were divided into four quartiles according to WBCC (WBCC1-4, from lowest to highest WBCC). The mean values of MetS components for each group were compared in males and females separately. Multivariate linear regression analysis between the WBCC and the components of MetS after adjusted for age and body mass index (BMI) were also evaluated. In the male group, the BMI of WBCC1 and WBCC2 was significantly lower than WBCC4. The total cholesterol and low-density lipoprotein-cholesterol (LDL-C) of WBCC2 were significantly higher than WBCC1 and WBCC4. Triglyceride (TG) levels of WBCC1 were significantly lower than WBCC3 and WBCC4, and TG levels of WBCC2 were significantly lower than WBCC4. Alternatively, the BMI of WBCC1 and WBCC2 were significantly lower than WBCC3 in the female group. Finally, the TG and fasting plasma glucose (FPG) levels of WBCC1 were significantly lower than WBCC3 or WBCC4, respectively. After multivariate linear regression, WBCC was positively correlated to BMI and TG, but negatively correlated to FPG in males whereas in young adolescent females, WBCC was positively correlated to BMI and FPG. In conclusion BMI was positively correlated with WBCC in young adolescent females and males. Thus, BMI is the most important component of MetS in this age group. In addition, TG levels in males and FPG in females were significantly related to WBCC. These findings could be regarded an early indication for the future development of full-blown MetS or cardiovascular diseases.

  20. Mammary pathogens and their relationship to somatic cell count and milk yield losses in dairy ewes.

    PubMed

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

    2002-06-01

    A total of 9592 samples of half udder milk were collected 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-Le6n region of Spain. Statistical analyses were carried out from a mixed model with random factor half udder or ewe for repeated measures. Test of significance of fixed effects of this mixed model showed significant effects of organisms, flock, parity, lactation stage, and birth type on SCC. Special reference must be made to novobiocin-sensitive coagulase-negative staphylococci, which represented more than 50% of the isolates and which elicited SCC geometric means of around 106/ml. In addition, the analysis of 4352 monthly test-day records for milk yield, SCC, and bacteriology showed that the ewes that were uninfected and infected by minor pathogens had the lowest SCC and the highest milk yields, whereas those infected by major pathogens had high SCC and milk yield losses between 8.8 and 10.1% according to the uni- or bilateral character of the infection. Finally, ewes infected by novobiocin-sensitive coagulase-negative staphylococci elicited SCC values similar to those of infections by major pathogens and milk yield losses ranging between those caused by minor and major pathogens. As a result, emphasis should be put on prevention of subclinical mastitis, particularly mastitis caused by novobiocin-sensitive coagulase-negative staphylococci in dairy sheep herds to improve microbiological and hygienic milk quality and to minimize losses in milk yield.

  1. Increased colostral somatic cell counts reduce pre-weaning calf immunity, health and growth.

    PubMed

    Ferdowsi Nia, E; Nikkhah, A; Rahmani, H R; Alikhani, M; Mohammad Alipour, M; Ghorbani, G R

    2010-10-01

    Our objective was to study the relationships between colostral somatic cell counts (SCC, a criterion for mastitis severity at parturition) and early calf growth, blood indicators of immunity, and pre-weaning faecal and health states. Sixty-nine Holstein cows were assigned to three groups of greater (n = 21, 5051 × 10(3)), medium (n = 38, 2138 × 10(3)) and lower (n = 10, 960 × 10(3)) colostral SCC (per ml) in a completely randomized design. Calves received 2 l of colostrum on day 1, and jugular blood was sampled at birth, at 3 h after the first colostrum feeding and at 42 days of age for immunoglobulin G (IgG) measurements. Calves were fed transition milk from their dams until 3 days of age and whole milk from 4 to 60 days of age twice daily at 10% of body weight. Health status and faecal physical scores were recorded daily for 42 days. Increased colostral SCC was associated with increased serum IgG at parturition. Colostral pH increased and fat percentage decreased linearly with the rising SCC. Feeding colostrum with greater SCC was associated with reduced serum IgG concentrations at 3 h after first colostrum feeding, greater incidences of diarrhoea and compromised health status during the first 42 days of age, and reduced weaning weight gain, but had no effects on calf body length and withers height. Colostral volume and percentages of protein, lactose, solids-non-fat, total solids and IgG were comparable among groups. Results suggest a role for SCC, as an indicator of mastitis and colostral health quality, in affecting calf health. As a result of the novelty of calf health dependence on colostral SCC found, future studies to further characterize such relationships and to uncover or rule out possible mediators are required before colostral SCC could be recommended for routine on-farm use in managing dry cow and calf production.

  2. Donor lymphocyte count and thymic activity predict lymphocyte recovery and outcomes after matched-sibling hematopoietic stem cell transplant.

    PubMed

    McIver, Zachariah; Melenhorst, Jan Joseph; Wu, Colin; Grim, Andrew; Ito, Sawa; Cho, Irene; Hensel, Nancy; Battiwalla, Minoo; Barrett, Austin John

    2013-03-01

    Delayed immune recovery is a characteristic feature of allogeneic hematopoietic stem cell transplantation in adult recipients. Although recipient thymic T-cell neogenesis contributes to T-cell regeneration after transplantation, thymic recovery in the transplant recipient decreases with increasing age, and is diminished by intensive preconditioning regimens and graft-versus-host disease. In adult recipients, most events that determine transplant success or failure occur during the period when the majority of circulating T cells is derived from the donor's post thymic T-cell repertoire. As a result, the make-up of the donor lymphocyte compartment may strongly influence immune recovery and transplant outcomes. The aim of this study was to examine donor lymphocyte counts in a series of patients undergoing an allogeneic hematopoietic stem cell transplant to identify the potential contribution of donor regulatory and conventional T lymphocyte populations to immune recovery and transplant outcomes. We examined donor lymphocyte subset counts in relation to post-transplant lymphocyte recovery and transplant events in 220 consecutive myeloablative, T-cell-depleted, HLA-identical sibling hematopoietic stem cell transplant recipients with hematologic malignancies. In a multivariate analysis, absolute numbers of donor CD4(+) recent thymic emigrants were associated with overall survival (P=0.032). The donors' absolute lymphocyte count and thymic production of regulatory T cells were both associated with extensive chronic graft-versus-host disease (P=0.002 and P=0.022, respectively). In conclusion, these results identify donor immune characteristics that are associated with lymphocyte recovery, extensive chronic graft-versus-host disease, and survival in the recipient following allogeneic hematopoietic stem cell transplantation. The study reported here was performed using peripheral blood samples drawn from donors and patients enrolled in the ClinicalTrials.gov-registered trials

  3. Counting mycobacteria in infected human cells and mouse tissue: a comparison between qPCR and CFU.

    PubMed

    Pathak, Sharad; Awuh, Jane A; Leversen, Nils Anders; Flo, Trude H; Asjø, Birgitta

    2012-01-01

    bacteria per host cell, an advantage in experiments where variable cell death can give misleading colony counts.

  4. CD4 cell count and CD4/CD8 ratio increase during rituximab maintenance in granulomatosis with polyangiitis patients

    PubMed Central

    Nossent, Johannes C.

    2016-01-01

    Introduction Rituximab (RTX) is a B cell-depleting agent approved for the treatment of granulomatosis with polyangiitis (GPA). RTX reduces antibody producing precursor plasma cells and inhibits B and T cells interaction. Infections related to T cell immunodeficiency are not infrequent during RTX treatment. Our study investigated CD4 cell count and CD4/CD8 ratio in GPA patients during the first two years of long-term RTX treatment. Methods A single centre cohort study of 35 patients who received median total cumulative dose of cyclophosphamide (CYC) of 15 g and were treated with RTX 2 g followed by retreatment with either 2 g once annually or 1 g biannually. Serum levels of total immunoglobulin (Ig) and lymphocytes subsets were recorded at RTX initiation and at 3, 6, 12, 18 and 24 months. Low CD4 count and inverted CD4/CD8 ratio were defined as CD4 < 0.3 × 109/l and ratio < 1. Results The CD4 cell count and CD4/CD8 ratio decreased slightly following the initial RTX treatment and then increased gradually during maintenance treatment. While the proportion of patients with low CD4 cell count decreased from 43% at baseline to 18% at 24 months, the ratio remained inverted in 40%. Oral daily prednisolone dose at baseline, CYC exposure and the maintenance regimen did not influence the CD4 cell count and ratio. Being older (p = 0.012) and having a higher CRP (p = 0.044) and ESR (p = 0.024) at baseline significantly increased the risk of inverted CD4/CD8 ratio at 24 months. Inverted ratio at baseline associated with lower total Ig levels during the study. Conclusions Overall, the CD4 and CD4/CD8 ratio increased during maintenance RTX therapy in GPA with no discernible impact of other immunosuppressive therapy. However the increase in CD4 was not followed by an increase in the CD4/CD8 ratio, especially in older patients. Inverted CD4/CD8 ratio associated with lower Ig levels, suggesting a more profound B cell depleting effect of RTX with a relative increase in CD8

  5. Accelerated CD4+ cell count decline in untreated HIV-1 patients points toward increasing virulence over the course of the epidemic.

    PubMed

    Brey, Florian L; Seybold, Ulrich; Kollan, Christian; Bogner, Johannes R

    2016-07-31

    Based on the assumption that the rate of CD4 cell count loss in treatment-naïve patients is correlated with the virulence of HIV-1, we evaluated 4616 patients. Patients who entered a German national database between 1985 and 1995 had a median annual CD4 cell count loss of 48 cells/μl, whereas those registered between 1999 and 2009 had a median annual CD4 cell count loss of 68 cells/μl (P < 0.001). This suggests that HIV-1 virulence has increased over the course of the epidemic.

  6. Preoperative lymphocyte count is a favorable prognostic factor of disease-free survival in non-small-cell lung cancer.

    PubMed

    Zhang, Jian; Huang, Shao-Hong; Li, Hui; Li, Yun; Chen, Xiu-Ling; Zhang, Wei-Qing; Chen, Hui-Guo; Gu, Li-Jia

    2013-03-01

    Recently, the prognostic value of cancer-related inflammatory response has been revealed. Previous studies showed that peripheral neutrophils and lymphocytes had significant impact on the prognosis of advanced and early-node-negative non-small-cell lung cancer (NSCLC). The purpose of this study was to investigate the prognostic value of preoperative lymphocyte and neutrophil counts in patients with NSCLC who underwent lobectomy and lymph node dissection and adjuvant chemotherapy. Retrospective analyses were performed to examine the impact of preoperative peripheral lymphocyte and neutrophil counts on disease-free survival (DFS) and overall survival (OS) and to analyze the relationships of these factors to clinicopathological factors. A total of 142 patients with NSCLC were evaluated of which 57 (40.1 %) patients had local recurrence or metastasis. Multivariate analyses revealed that peripheral lymphocyte count was an independent favorable prognostic factor of DFS (hazard ratio 0.548; 95 % confidence interval 0.351-0.857; P = 0.008) but not OS (P = 0.164). The maximum logrank statistical value was 9.504 (P = 0.002) when the cutoff value of lymphocyte was 1,800 mm(-3). The median DFS was 318.0 days (95 % confidence interval 226.0-410.0) for lymphocyte ≤1,800 mm(-3) group and 669.0 days (95 % confidence interval 0.0-1,431.0) for lymphocyte >1,800 mm(-3) group. Low lymphocyte count was related with lymphatic invasion (P = 0.012) and recurrence of NSCLC (P = 0.022). Peripheral neutrophil count had no impact on DFS or OS when analysis included all the 142 patients. Preoperative peripheral lymphocyte count, which is related with lymphatic invasion, is an independent favorable prognostic factor of DFS in patients with NSCLC who underwent lobectomy and lymph node dissection and adjuvant chemotherapy.

  7. Value of counting colonic mucosal Ig-containing cells in the differential diagnosis of chronic inflammatory bowel disease.

    PubMed Central

    Seldenrijk, C. A.; Meuwissen, S. G.; Schipper, N. W.; Morson, B. C.; Lindeman, J.; Meijer, C. J.

    1992-01-01

    AIMS: To investigate whether counting cells containing immunoglobulin (Ig) subclass in colonic biopsy specimens of patients with chronic inflammatory bowel disease, in addition to conventional histological evaluation, can improve the differentiation of patients with Crohn's disease from those with ulcerative colitis. METHODS: The colonic and rectal biopsy specimens of 40 patients with chronic inflammatory bowel disease, comprising 20 patients with Crohn's disease and 20 with ulcerative colitis, were used and sections were stained specifically for IgA, IgM, and IgG heavy chains using an indirect immune peroxidase method. The immunoglobulin subclass containing cells were counted using an ocular grid counting method in a light microscope. A linear stepwise discriminant analysis was performed on Ig subclass containing cell counts in combination with 16 reproducible histological features. The results of this discriminant analysis were compared with the results of the discriminant analyses in which only the histological features were used. RESULTS: Applying stepwise discriminant analysis, two histological features (an excess of histiocytes in the lamina propria and the villous or irregular aspect of the mucosal surface) in combination with IgMax were selected as the most discriminatory parameters that distinguish Crohn's disease from ulcerative colitis. IgMmax was defined as the maximum value of the mean percentage of IgM containing cells over all the biopsy locations. The use of this combination resulted in a better classification in 20% of the patients with Crohn's disease and in 9% of the patients with ulcerative colitis compared with the use of histological features alone. CONCLUSIONS: Morphometric enumeration of Ig subclass containing cells in colonic mucosal biopsy specimens has diagnostic value as a means of differentiating individual patients with Crohn's disease from those with ulcerative colitis. PMID:1556234

  8. Ultrasound-detected joint inflammation and B cell count: related variables for rituximab-treated RA patients?

    PubMed

    Valor, Lara; Martínez-Estupiñán, Lina; Janta, Iustina; Nieto, Juan Carlos; Ovalles-Bonilla, Juan Gabriel; González-Fernández, Carlos; Del Rio, Tamara; Hernández-Flórez, Diana; Monteagudo, Indalecio; López-Longo, Francisco Javier; Naredo, Esperanza

    2016-06-01

    This cross-sectional observational study aimed to explore the relationship between B cell count and ultrasound (US)-detected synovitis, in patients with rheumatoid arthritis treated with rituximab. Thirty-seven consecutive RA patients treated with RTX were recruited for the study. The patients underwent clinical [i.e., Disease Activity Score 28 joints (DAS28)], laboratory, and US assessment of 12 joints. Each joint was semiquantitatively (0-3) scored on B-mode and power Doppler mode. The scores were summed, and a global index was created for BM (BMS) and PD scores (PDI) synovitis. BM subclinical synovitis was evident in all patients, with PD synovial signal detected in 16 patients (43.2 %). No correlation was found between DAS28 and US scores. B cells were detected in 27 (72.9 %) patients, but there was no association in the mean B cell count and disease activity as measured by DAS28 (DAS28 < 2.6 = 34.53, DAS28 > 2.6 = 49.45, p = 0.52) and PDI score (PDI < 1 = 49.48, PDI > 1 = 35.44, p = 0.54). There was no correlation between the B cell count and DAS28, BMS, and PDI (r = 0.020, p = 0.907; r = -0.151, p = 0.371; r = -0.099, p = 0.558, respectively). In RTX-treated RA patients, no relationship could be established between US-detected synovitis and peripheral blood B cell count.

  9. Quantification of cells with specific phenotypes I: determination of CD4+ cell count per microliter in reconstituted lyophilized human PBMC prelabeled with anti-CD4 FITC antibody.

    PubMed

    Stebbings, Richard; Wang, Lili; Sutherland, Janet; Kammel, Martin; Gaigalas, Adolfas K; John, Manuela; Roemer, Bodo; Kuhne, Maren; Schneider, Rudolf J; Braun, Michael; Engel, Andrea; Dikshit, Dinesh K; Abbasi, Fatima; Marti, Gerald E; Sassi, Maria Paola; Revel, Laura; Kim, Sook-Kyung; Baradez, Marc-Olivier; Lekishvili, Tamara; Marshall, Damian; Whitby, Liam; Jing, Wang; Ost, Volker; Vonsky, Maxim; Neukammer, Jörg

    2015-03-01

    A surface-labeled lyophilized lymphocyte (sLL) preparation has been developed using human peripheral blood mononuclear cells prelabeled with a fluorescein isothiocyanate conjugated anti-CD4 monoclonal antibody. The sLL preparation is intended to be used as a reference material for CD4+ cell counting including the development of higher order reference measurement procedures and has been evaluated in the pilot study CCQM-P102. This study was conducted across 16 laboratories from eight countries to assess the ability of participants to quantify the CD4+ cell count of this reference material and to document cross-laboratory variability plus associated measurement uncertainties. Twelve different flow cytometer platforms were evaluated using a standard protocol that included calibration beads used to obtain quantitative measurements of CD4+ T cell counts. There was good overall cross-platform and counting method agreement with a grand mean of the laboratory calculated means of (301.7 ± 4.9) μL(-1) CD4+ cells. Excluding outliers, greater than 90% of participant data agreed within ±15%. A major contribution to variation of sLL CD4+ cell counts was tube to tube variation of the calibration beads, amounting to an uncertainty of 3.6%. Variation due to preparative steps equated to an uncertainty of 2.6%. There was no reduction in variability when data files were centrally reanalyzed. Remaining variation was attributed to instrument specific differences. CD4+ cell counts obtained in CCQM-P102 are in excellent agreement and show the robustness of both the measurements and the data analysis and hence the suitability of sLL as a reference material for interlaboratory comparisons and external quality assessment.

  10. Quantification of Cells with Specific Phenotypes I: Determination of CD4+ Cell Count Per Microliter in Reconstituted Lyophilized Human PBMC Prelabeled with Anti-CD4 FITC Antibody

    PubMed Central

    Stebbings, Richard; Wang, Lili; Sutherland, Janet; Kammel, Martin; Gaigalas, Adolfas K; John, Manuela; Roemer, Bodo; Kuhne, Maren; Schneider, Rudolf J; Braun, Michael; Engel, Andrea; Dikshit, Dinesh K; Abbasi, Fatima; Marti, Gerald E; Paola Sassi, Maria; Revel, Laura; Kim, Sook-Kyung; Baradez, Marc-Olivier; Lekishvili, Tamara; Marshall, Damian; Whitby, Liam; Jing, Wang; Ost, Volker; Vonsky, Maxim; Neukammer, Jörg

    2015-01-01

    A surface-labeled lyophilized lymphocyte (sLL) preparation has been developed using human peripheral blood mononuclear cells prelabeled with a fluorescein isothiocyanate conjugated anti-CD4 monoclonal antibody. The sLL preparation is intended to be used as a reference material for CD4+ cell counting including the development of higher order reference measurement procedures and has been evaluated in the pilot study CCQM-P102. This study was conducted across 16 laboratories from eight countries to assess the ability of participants to quantify the CD4+ cell count of this reference material and to document cross-laboratory variability plus associated measurement uncertainties. Twelve different flow cytometer platforms were evaluated using a standard protocol that included calibration beads used to obtain quantitative measurements of CD4+ T cell counts. There was good overall cross-platform and counting method agreement with a grand mean of the laboratory calculated means of (301.7 ± 4.9) μL−1 CD4+ cells. Excluding outliers, greater than 90% of participant data agreed within ±15%. A major contribution to variation of sLL CD4+ cell counts was tube to tube variation of the calibration beads, amounting to an uncertainty of 3.6%. Variation due to preparative steps equated to an uncertainty of 2.6%. There was no reduction in variability when data files were centrally reanalyzed. Remaining variation was attributed to instrument specific differences. CD4+ cell counts obtained in CCQM-P102 are in excellent agreement and show the robustness of both the measurements and the data analysis and hence the suitability of sLL as a reference material for interlaboratory comparisons and external quality assessment. © 2015 The Authors. Published by Wiley Periodicals, Inc. PMID:25655255

  11. Low plasma adiponectin level, white blood cell count and Helicobacter pylori titre independently predict abnormal pancreatic beta-cell function.

    PubMed

    So, Wing-Yee; Tong, Peter C; Ko, Gary T; Ma, Ronald C; Ozaki, Risa; Kong, Alice P; Yang, Xilin; Ho, Chung-Shun; Lam, Christopher C; Chan, Juliana C

    2009-11-01

    Adiponectin is an adipocytokine with insulin sensitizing effect while chronic inflammation damages pancreatic beta-cells leading to reduced insulin response. We aimed to prove the hypothesis that adiponectin levels and inflammatory markers (white blood cell counts [WCC], Helicobacter pylori [HP] titers, high sensitivity C-reactive protein [hs-CRP]) may interact to affect risk of diabetes. We studied 288 Chinese men (age-median: 41.0 years, IQR: 35.3-46.0 years) being recruited from the community in Hong Kong. The mean adiponectin level was 5.39+/-2.81 microg/ml and 40.9% (n=107) had low adiponectin level (<4 microg/ml). On multiple regression analysis, adiponectin was negatively associated with diabetes, HOMA insulin resistance top quartile, plasma glucose (PG) and 2h insulin; and positively associated with HOMA insulin sensitivity index. WCC was independently associated with PG and 15' insulin, and negatively associated with HOMA insulin sensitivity top quartile. HP titre was associated with 30' PG level and diabetes. hs-CRP did not enter the multivariable model. In conclusion, adiponectin, WCC and HP titer are independent predictors for hyperglycemia and reduced insulin sensitivity in Chinese men. These findings may explain the high risk for diabetes in Chinese population despite their relatively low adiposity.

  12. Clinical utility of circulating tumor cell counting through CellSearch®: the dilemma of a concept suspended in Limbo

    PubMed Central

    Raimondi, Cristina; Gradilone, Angela; Naso, Giuseppe; Cortesi, Enrico; Gazzaniga, Paola

    2014-01-01

    To date, 10 years after the first demonstration of circulating tumor cells (CTCs), prognostic significance in metastatic breast cancer using the US Food and Drug Administration–cleared system CellSearch®, the potential utility of CTCs in early clinical development of drugs, their role as a surrogate marker of response to therapy, and their molecular analysis for patient stratification for targeted therapies are still major unsolved questions. Great expectations are pinned on the ongoing interventional trials aimed to demonstrate that CTCs might be of value for guiding treatment of patients and predicting cancer progression. To fill the gap between theory and practice with regard to the clinical utility of CTCs, a bridge is needed, taking into account innovative design for clinical trials, a revised definition of traditional CTCs, next-generation CTC technology, the potential clinical application of CTC analysis in non-validated settings of disease, and finally, expanding the number of patients enrolled in the studies. In this regard, the results of the first European pooled analysis definitely validated the independent prognostic value of CTC counting in metastatic breast cancer patients. PMID:24790460

  13. Cytotoxicity of selected magnetic fluids on human adenocarcinoma cells

    NASA Astrophysics Data System (ADS)

    Hilger, Ingrid; Frühauf, Sylvia; Linß, Werner; Hiergeist, Robert; Andrä, Wilfried; Hergt, Rudolf; Kaiser, Werner A.

    2003-04-01

    Based on the knowledge that the magnetite particles seem to be well tolerated by the human body, the cytotoxic potential of coated particles was investigated, which had been selected for potential applications regarding the minimal-invasive elimination of breast tumors by magnetic thermoablation. Human adenocarcinoma cells (BT-20) were exposed (24, 48 and 72 h) to different magnetite particles with diverging total size (8, 10 and 220 nm) and coating (cationic and anionic). One sample contained only non-coated magnetite particles. The magnetite concentration ranged between 0.2 and 20 ng/cell. Cytotoxicity was estimated by measuring the succinate dehydrogenase activity. The morphologic features resulting from the interaction of magnetic fluids with BT-20 cells was determined by transmission electron microscopy. As opposed to the non-coated magnetic particles, cationic particles induced the strongest decrease in cell survival rates depending on time and concentration. Morphologically, the cationic particle samples exerted a strong binding to cellular membranes. Changes in the subcellular structure were found in relation to the coated magnetic particles. In conclusion, our results show that the coated prototype magnetic particles, particularly those with a cationic surfactant, are cytotoxic to BT-20 cells. The cytotoxicity is attributed to electrostatic bindings with cellular membranes, influences of chemical components or non-physiologic pH. Considering the in vivo applications, adverse systemic effects are conceivable and more biocompatible coatings for the selected magnetic particles should be elaborated.

  14. Cell type specific DNA methylation in cord blood: A 450K-reference data set and cell count-based validation of estimated cell type composition.

    PubMed

    Gervin, Kristina; Page, Christian Magnus; Aass, Hans Christian D; Jansen, Michelle A; Fjeldstad, Heidi Elisabeth; Andreassen, Bettina Kulle; Duijts, Liesbeth; van Meurs, Joyce B; van Zelm, Menno C; Jaddoe, Vincent W; Nordeng, Hedvig; Knudsen, Gunn Peggy; Magnus, Per; Nystad, Wenche; Staff, Anne Cathrine; Felix, Janine F; Lyle, Robert

    2016-09-01

    Epigenome-wide association studies of prenatal exposure to different environmental factors are becoming increasingly common. These studies are usually performed in umbilical cord blood. Since blood comprises multiple cell types with specific DNA methylation patterns, confounding caused by cellular heterogeneity is a major concern. This can be adjusted for using reference data consisting of DNA methylation signatures in cell types isolated from blood. However, the most commonly used reference data set is based on blood samples from adult males and is not representative of the cell type composition in neonatal cord blood. The aim of this study was to generate a reference data set from cord blood to enable correct adjustment of the cell type composition in samples collected at birth. The purity of the isolated cell types was very high for all samples (>97.1%), and clustering analyses showed distinct grouping of the cell types according to hematopoietic lineage. We explored whether this cord blood and the adult peripheral blood reference data sets impact the estimation of cell type composition in cord blood samples from an independent birth cohort (MoBa, n = 1092). This revealed significant differences for all cell types. Importantly, comparison of the cell type estimates against matched cell counts both in the cord blood reference samples (n = 11) and in another independent birth cohort (Generation R, n = 195), demonstrated moderate to high correlation of the data. This is the first cord blood reference data set with a comprehensive examination of the downstream application of the data through validation of estimated cell types against matched cell counts.

  15. Red Blood Cell Distribution Width and the Platelet Count in Iron-deficient Children Aged 0.5-3 Years.

    PubMed

    Akkermans, M D; Uijterschout, L; Vloemans, J; Teunisse, P P; Hudig, F; Bubbers, S; Verbruggen, S; Veldhorst, M; de Leeuw, T G; van Goudoever, J B; Brus, F

    2015-01-01

    Early detection of iron deficiency (ID) and iron deficiency anemia (IDA) in young children is important to prevent impaired neurodevelopment. Unfortunately, many biomarkers of ID are influenced by infection, thus limiting their usefulness. The aim of this study was to investigate the value of red blood cell distribution width (RDW) and the platelet count for detecting ID(A) among otherwise healthy children. A multicenter prospective observational study was conducted in the Netherlands to investigate the prevalence of ID(A) in 400 healthy children aged 0.5-3 years. ID was defined as serum ferritin (SF) <12 μg/L in the absence of infection (C-reactive protein [CRP] <5 mg/L) and IDA as hemoglobin <110 g/L combined with ID. RDW (%) and the platelet count were determined in the complete blood cell count. RDW was inversely correlated with SF and not associated with CRP. Calculated cutoff values for RDW to detect ID and IDA gave a relatively low sensitivity (53.1% and 57.1%, respectively) and specificity (64.7% and 69.9%, respectively). Anemic children with a RDW >14.3% had a 2.7 higher odds (95% confidence interval [CI]: 1.2-6.3) to be iron deficient, compared with anemic children with a RDW <14.3%. The platelet count showed a large range in both ID and non-ID children. In conclusion, RDW can be helpful for identifying ID as the cause of anemia in 0.5- to 3-year-old children, but not as primary biomarker of ID(A). RDW values are not influenced by the presence of infection. There appears to be no role for the platelet count in diagnosing ID(A) in this group of children.

  16. White Blood Cell Counts as Risk Markers of Developing Metabolic Syndrome and Its Components in the Predimed Study

    PubMed Central

    Babio, Nancy; Ibarrola-Jurado, Núria; Bulló, Mònica; Martínez-González, Miguel Ángel; Wärnberg, Julia; Salaverría, Itziar; Ortega-Calvo, Manuel; Estruch, Ramón; Serra-Majem, Lluís; Covas, Maria Isabel; Sorli, José Vicente; Salas-Salvadó, Jordi

    2013-01-01

    Background The Metabolic Syndrome (MetS) is a cluster of metabolic abnormalities that includes hyperglucemia, hypertension, dyslipidemia and central obesity, conferring an increased risk of cardiovascular disease. The white blood cell (WBC) count has been proposed as a marker for predicting cardiovascular risk. However, few prospective studies have evaluated the relationship between WBC subtypes and risk of MetS. Methods Participants were recruited from seven PREDIMED study centers. Both a baseline cross-sectional (n = 4,377) and a prospective assessment (n = 1,637) were performed. Participants with MetS at baseline were excluded from the longitudinal analysis. The median follow-up was 3.9 years. Anthropometric measurements, blood pressure, fasting glucose, lipid profile and WBC counts were assessed at baseline and yearly during the follow-up. Participants were categorized by baseline WBC and its subtype count quartiles. Adjusted logistic regression models were fitted to assess the risk of MetS and its components. Results Of the 4,377 participants, 62.6% had MetS at baseline. Compared to the participants in the lowest baseline sex-adjusted quartile of WBC counts, those in the upper quartile showed an increased risk of having MetS (OR, 2.47; 95%CI, 2.03–2.99; P-trend<0.001). This association was also observed for all WBC subtypes, except for basophils. Compared to participants in the lowest quartile, those in the top quartile of leukocyte, neutrophil and lymphocyte count had an increased risk of MetS incidence. Leukocyte and neutrophil count were found to be strongly associated with the MetS components hypertriglyceridemia and low HDL-cholesterol. Likewise, lymphocyte counts were found to be associated with the incidence of the MetS components low HDL-cholesterol and high fasting glucose. An increase in the total WBC during the follow-up was also associated with an increased risk of MetS. Conclusions Total WBC counts, and some subtypes, were positively

  17. Tumor infiltrating CD8(+) T lymphocyte count is independent of tumor TLR9 status in treatment naïve triple negative breast cancer and renal cell carcinoma.

    PubMed

    Mella, Mikko; Kauppila, Joonas H; Karihtala, Peeter; Lehenkari, Petri; Jukkola-Vuorinen, Arja; Soini, Ylermi; Auvinen, Päivi; Vaarala, Markku H; Ronkainen, Hanna; Kauppila, Saila; Haapasaari, Kirsi-Maria; Vuopala, Katri S; Selander, Katri S

    2015-06-01

    Toll-like receptor 9 (TLR9) is a cellular DNA-receptor of the innate immune system that is widely expressed in cancers. We demonstrated that low tumor TLR9 expression predicts poor disease-specific survival in triple negative breast cancer (TNBC) and renal cell carcinoma (RCC). We hypothesized that this is because TLR9 expression affects tumor immunophenotype. To begin to test this, we compared the number of tumor infiltrating CD8(+) T lymphocytes with TLR9 expression in treatment naïve breast cancer (n = 197) and RCC (n = 94) cohorts with known TLR9 expression status. CD8(+) T lymphocyte counts were assayed with image analysis after immunohistochemistry (IHC). Tumor TLR9 expression was not correlated with CD8(+) T cell counts in breast cancer or RCC. CD8(+) T cell counts were significantly associated with tumor proliferation index in TNBC, but not in non-TNBC. CD8(+) T cell counts were also significantly associated with tumor grade in non-TNBC, but not in TNBC. In RCC, CD8(+) T cell counts were significantly associated with tumor stage. CD8(+) T cell counts were significantly associated with prognosis in TNBC and RCC, but the presence of CD8(+) T cells in these tumors had opposite effects on disease-specific survival: High CD8(+) counts were associated with better prognosis in TNBC and worse prognosis in RCC. Among TNBC patients, those with low tumor TLR9 and low CD8(+) T cell counts had the poorest prognosis (log-rank p = 0.0002 vs. high tumor TLR9 and high CD8(+) T cell count). In conclusion, pre-treatment tumor TLR9 status is not associated with tumor infiltrating CD8(+) T lymphocytes in TNBC or RCC. The combination of TLR9 and CD8(+) TIL count might be a novel composite prognostic marker in TNBC.

  18. Effect of somatic cell count in goat milk on yield, sensory quality, and fatty acid profile of semisoft cheese.

    PubMed

    Chen, S X; Wang, J Z; Van Kessel, J S; Ren, F Z; Zeng, S S

    2010-04-01

    This study investigated the effect of somatic cell count (SCC) in goat milk on yield, free fatty acid (FFA) profile, and sensory quality of semisoft cheese. Sixty Alpine goats without evidence of clinical mastitis were assigned to 3 groups with milk SCC level of <500,000 (low), 500,000 to 1,000,000 (medium), and 1,000,000 to 1,500,000 (high) cells/mL. Thirty kilograms of goat milk with mean SCC levels of 410,000 (low), 770,000 (medium), and 1,250,000 (high) cells/mL was obtained for the manufacture of semisoft cheese for 2 consecutive weeks in 3 lactation stages. The composition of milk was analyzed and cheese yield was recorded on d 1. Cheese samples on d 1, 60, and 120 were analyzed for total sensory scores, flavor, and body and texture by a panel of 3 expert judges and were also analyzed for FFA. Results indicated that milk composition did not change when milk SCC varied from 214,000 to 1,450,000 cells/mL. Milk with higher SCC had a lower standard plate count, whereas coliform count and psychrotrophic bacteria count were not affected. However, milk components (fat, protein, lactose, casein, and total solids) among the 3 groups were similar. As a result, no significant differences in the yield of semisoft goat cheeses were detected. However, total sensory scores and body and texture scores for cheeses made from the high SCC milk were lower than those for cheeses made from the low and medium SCC milks. The difference in milk SCC levels also resulted in diverse changes in cheese texture (hardness, springiness, and so on) and FFA profiles. Individual and total FFA increased significantly during ripening, regardless the SCC levels. It is concluded that SCC in goat milk did not affect the yield of semisoft cheese but did result in inferior sensory quality of aged cheeses.

  19. Composition, proteolysis indices and coagulating properties of ewe milk as affected by bulk tank somatic cell count.

    PubMed

    Martí-De Olives, Ana; Navarro-Ríos, María Jesús; Rubert-Alemán, Joaquín; Fernández, Nemesio; Molina, Maria Pilar

    2015-08-01

    The aim of this study was to assess the effect of ovine bulk tank somatic cell count (BTSCC) on composition, proteose-peptone (p-p) content and casein fractions as indicating parameters for proteolysis and coagulating properties of milk. A total of 97 samples of bulk tank milk from Manchega breed ewe flocks were grouped according to somatic cell count (SCC) into four classes: fewer than 500,000 cells/ml, from 500,000 to 10,00000 cells/ml, from 10,00000 to 15,00000 and more than 15,00000 cells/ml. The casein : protein ratio and lactose content decreased with BTSCC. Proteolysis increased with BTSCC, causing a drop in β-casein and an increase in the γ-caseins from a concentration of 500,000 cells/ml. Regarding coagulation behaviour, the rennet clotting time (RCT) and firming time (k20) rose from 10,00000-15,00000 cells/ml of milk. The results showed that the impairment of milk quality and milk ability to make cheese as affected by intramammary infection (IMI) can be inferred from the bulk tank milk of flocks with poor udder health.

  20. Interactions between cadmium and decabrominated diphenyl ether on blood cells count in rats-Multiple factorial regression analysis.

    PubMed

    Curcic, Marijana; Buha, Aleksandra; Stankovic, Sanja; Milovanovic, Vesna; Bulat, Zorica; Đukić-Ćosić, Danijela; Antonijević, Evica; Vučinić, Slavica; Matović, Vesna; Antonijevic, Biljana

    2017-02-01

    The objective of this study was to assess toxicity of Cd and BDE-209 mixture on haematological parameters in subacutely exposed rats and to determine the presence and type of interactions between these two chemicals using multiple factorial regression analysis. Furthermore, for the assessment of interaction type, an isobologram based methodology was applied and compared with multiple factorial regression analysis. Chemicals were given by oral gavage to the male Wistar rats weighing 200-240g for 28days. Animals were divided in 16 groups (8/group): control vehiculum group, three groups of rats were treated with 2.5, 7.5 or 15mg Cd/kg/day. These doses were chosen on the bases of literature data and reflect relatively high Cd environmental exposure, three groups of rats were treated with 1000, 2000 or 4000mg BDE-209/kg/bw/day, doses proved to induce toxic effects in rats. Furthermore, nine groups of animals were treated with different mixtures of Cd and BDE-209 containing doses of Cd and BDE-209 stated above. Blood samples were taken at the end of experiment and red blood cells, white blood cells and platelets counts were determined. For interaction assessment multiple factorial regression analysis and fitted isobologram approach were used. In this study, we focused on multiple factorial regression analysis as a method for interaction assessment. We also investigated the interactions between Cd and BDE-209 by the derived model for the description of the obtained fitted isobologram curves. Current study indicated that co-exposure to Cd and BDE-209 can result in significant decrease in RBC count, increase in WBC count and decrease in PLT count, when compared with controls. Multiple factorial regression analysis used for the assessment of interactions type between Cd and BDE-209 indicated synergism for the effect on RBC count and no interactions i.e. additivity for the effects on WBC and PLT counts. On the other hand, isobologram based approach showed slight antagonism

  1. Impact of ambient air pollution on the differential white blood cell count in patients with chronic pulmonary disease.

    PubMed

    Brüske, Irene; Hampel, Regina; Socher, Martin M; Rückerl, Regina; Schneider, Alexandra; Heinrich, Joachim; Oberdörster, Günter; Wichmann, H-Erich; Peters, Annette

    2010-02-01

    Epidemiologic studies report associations between particulate air pollution and increased mortality from pulmonary diseases. This study was performed to examine whether the exposure to ambient gaseous and particulate air pollution leads to an alteration of the differential white blood cell count in patients with chronic pulmonary diseases like chronic bronchitis, chronic obstructive pulmonary disease, and asthma. A prospective panel study was conducted in Erfurt, Eastern Germany, with 12 repeated differential white blood cell counts in 38 males with chronic pulmonary diseases. Hourly particulate and gaseous air pollutants and meteorological data were acquired. Mixed models with a random intercept adjusting for trend, meteorology, weekday, and other risk variables were used. In this explorative analysis, we found an immediate decrease of polymorphonuclear leukocytes in response to an increase of most gaseous and particulate pollutants. Lymphocytes increased within 24 h in association with all gaseous pollutants but showed only minor effects in regard to particulate air pollution. Monocytes showed an increase associated with ultrafine particles, and nitrogen monoxide. The effect had two peaks in time, one 0-23 h before blood withdrawal and a second one with a time lag of 48-71 h. The increase of particulate and gaseous air pollution was associated with multiple changes in the differential white blood cell count in patients with chronic pulmonary diseases.

  2. Identification of cell morphology parameters from automatic hematology analyzers to predict the peripheral blood CD34-positive cell count after mobilization

    PubMed Central

    Shin, Saeam; Cho, Sung Ran; Kim, Sinyoung; Choi, Jong Rak; Lee, Kyung-A

    2017-01-01

    Optimal timing of apheresis initiation is important for maximizing the hematopoietic stem cell (HSC) yield. This study aimed to identify useful parameters from automatic hematology analyzers for predicting the peripheral blood CD34+ cell count after mobilization. We prospectively enrolled 53 healthy donors and 72 patients, and evaluated 43 cell morphology parameters from Unicel DxH800 (Beckman Coulter, USA) and Advia 2120i (Siemens Healthcare Diagnostics, USA). The correlation of each parameter with the CD34+ cell count in pre-apheresis blood samples was analyzed. The delta neutrophil index (DNI) from Advia 2120i, standard deviation of volume of neutrophils and monocytes (SD-V-NE and SD-V-MO), standard deviation of conductivity of neutrophils and monocytes (SD-C-NE and SD-C-MO), mean conductivity of neutrophils and monocytes (MN-C-NE and MN-C-MO), and standard deviation of axial light loss of neutrophils and monocytes (SD-AL2-NE and SD-AL2-MO) from DxH800 showed significant correlations with the CD34+ cell count. SD-V-NE, SD-C-NE, and SD-C-MO showed good or fair area under the curve values for the prediction of the CD34+ cell count. SD-V-NE, SD-C-NE, and SD-C-MO from DxH800 will provide rapid, useful information for the initiation of apheresis after mobilization. PMID:28319139

  3. Estimates of genetic parameters for daily somatic cell count of Australian dairy cattle.

    PubMed

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

    2001-05-01

    Genetic parameters for daily somatic cell counts (SCC) of the first three parities were estimated for Australian Dairy Cattle. Most of the data analyses were carried out with a sire random regression model. The estimates were compared with those from conventional ten-trait analyses and animal models. In the first-parity estimates of heritabilities (h2) were low (0.04 to 0.05) at the beginning of the lactation and higher (0.11 to 0.13) at the end. The average h2 estimated from random regression sire model, random regression animal model and conventional multitrait sire model were 0.09, 0.09, and 0.08, respectively, in the first lactation. The average h2 were 0.09 and 0.11 in the second and third parities, respectively. Genetic correlations between daily log(e) SCC within parity were high for adjacent tests (nearly 1) and low (as low as 0.30) between the beginning and the end of the lactation. Generally, the genetic correlations between parities depend on how far apart they are and on whether they are on the same day in any two parities. Across parities, on average, genetic correlations between parities 1 and 3 were the lowest and those between 1 and 2 intermediate, while those between 2 and 3 were the highest. The estimated environmental correlations were lower than the genetic correlations, but the trends were generally similar. Differences in genetic parameter estimates due to model were small, except for some genetic correlations. The high residual error variances, the low h2, and the inconsistency in genetic correlations that were observed particularly at the beginning of the first lactation suggest that log(e) SCC early in the first lactation may be related to a spike in SCC as result of infection and (or) onset of lactation while SCC later in lactation represents a sustained response to infection. Accounting for the variation in heritabilities and correlations should improve the accuracy of genetic evaluations for SCC based on test day records.

  4. 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

  5. Differentiation of mesenchymal stem cells from human amniotic fluid to vascular endothelial cells.

    PubMed

    Tancharoen, Waleephan; Aungsuchawan, Sirinda; Pothacharoen, Peraphan; Markmee, Runchana; Narakornsak, Suteera; Kieodee, Junjira; Boonma, Nonglak; Tasuya, Witoon

    2017-03-01

    Endothelial dysfunction is a principle feature of vascular-related disease. Endothelial cells have been acquired for the purposes of the restoration of damaged tissue in therapeutic angiogenesis. However, their use is limited by expansion capacity and the small amount of cells that are obtained. Human amniotic fluid mesenchymal stem cells (hAF-MSCs) are considered an important source for vascular tissue engineering. In this study, hAF-MSCs were characterized and then induced in order to differentiate into the endothelial-like cells. Human amniotic fluid cells (hAFCs) were obtained from amniocentesis at the second trimester of gestation. The cells were characterized as mesenchymal stem cells by flow cytometry. The results showed that the cells were positive for mesenchymal stem cell markers CD44, CD73, CD90 and HLA-ABC, and negative for CD31, Amniotic fluid stem cells marker: CD117, anti-human fibroblasts, HLA-DR and hematopoietic differentiation markers CD34 and CD45. The hAF-MSCs were differentiated into endothelial cells under the induction of vascular endothelial growth factor (VEGF) and analyzed for the expression of the endothelial-specific markers and function. The expression of the endothelial-specific markers was determined by reverse transcriptase-quantitative PCR (RT-qPCR), while immunofluorescent analysis demonstrated that the induced hAF-MSCs expressed von Willebrand factor (vWF), vascular endothelial growth factor receptor 2 (VEGFR2), CD31 and endothelial nitric oxide synthase (eNOS). The network formation assay showed that the induced hAF-MSCs formed partial networks. All results indicated that hAF-MSCs have the potential to be differentiated into endothelial-like cells, while human amniotic fluid might be a suitable source of MSCs for vascularized tissue engineering.

  6. Vγ9Vδ2 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 γδ 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-Vγ9Vδ2 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 Vγ9Vδ2-polyfunctional response quality is affected, since several Vγ9Vδ2 T-cell subsets resulted significantly lower in HIV+ patients in respect to healthy donors. Interestingly, we found a weak positive correlation between Vγ9Vδ2 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 Vγ9Vδ2 T-cell subsets expressing MIP-1β in different combinations with other molecules (CD107a/IFNγ) in respect to High-CD4 individuals. Our results show that the Vγ9Vδ2 T-cell-response quality in Low-CD4 patients is specifically affected, suggesting a direct link between innate Vγ9Vδ2 T-cells and CD4 T-cell count. These findings suggest that Vγ9Vδ2 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

  7. Vγ9Vδ2 T-Cell Polyfunctionality Is Differently Modulated in HAART-Treated HIV Patients according to CD4 T-Cell Count.

    PubMed

    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 γδ 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-Vγ9Vδ2 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 Vγ9Vδ2-polyfunctional response quality is affected, since several Vγ9Vδ2 T-cell subsets resulted significantly lower in HIV+ patients in respect to healthy donors. Interestingly, we found a weak positive correlation between Vγ9Vδ2 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 Vγ9Vδ2 T-cell subsets expressing MIP-1β in different combinations with other molecules (CD107a/IFNγ) in respect to High-CD4 individuals. Our results show that the Vγ9Vδ2 T-cell-response quality in Low-CD4 patients is specifically affected, suggesting a direct link between innate Vγ9Vδ2 T-cells and CD4 T-cell count. These findings suggest that Vγ9Vδ2 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.

  8. Physical activity, white blood cell count, and lung cancer risk in a prospective cohort study.

    PubMed

    Sprague, Brian L; Trentham-Dietz, Amy; Klein, Barbara E K; Klein, Ronald; Cruickshanks, Karen J; Lee, Kristine E; Hampton, John M

    2008-10-01

    Previous studies have suggested that physical activity may lower lung cancer risk. The association of physical activity with reduced chronic inflammation provides a potential mechanism, yet few studies have directly related inflammatory markers to cancer incidence. The relation among physical activity, inflammation, and lung cancer risk was evaluated in a prospective cohort of 4,831 subjects, 43 to 86 years of age, in Beaver Dam, Wisconsin. A total physical activity index was created by summing up kilocalories per week from sweat-inducing physical activities, city blocks walked, and flights of stairs climbed. Two inflammatory markers, WBC count and serum albumin, were measured at the baseline examination. During an average of 12.8 years of follow-up, 134 incident cases of lung cancer were diagnosed. After multivariable adjustment, participants in the highest tertile of total physical activity index had a 45% reduction in lung cancer risk compared with those in the lowest tertile (hazard ratio, 0.55; 95% confidence interval, 0.35-0.86). Participants with WBC counts in the upper tertile (>or=8 x 10(3)/microL) were 2.81 (95% confidence interval, 1.58-5.01) times as likely to develop lung cancer as those with counts in the lowest tertile (<6.4 x 10(3)/microL). Serum albumin was not related to lung cancer risk. There was no evidence that inflammation mediated the association between physical activity and lung cancer risk, as the physical activity risk estimates were essentially unchanged after adjustment for WBC count. Although the potential for residual confounding by smoking could not be eliminated, these data suggest that physical activity and WBC count are independent risk factors for lung cancer.

  9. Why translation counts for mitochondria - retrograde signalling links mitochondrial protein synthesis to mitochondrial biogenesis and cell proliferation.

    PubMed

    Battersby, Brendan J; Richter, Uwe

    2013-10-01

    Organelle biosynthesis is a key requirement for cell growth and division. The regulation of mitochondrial biosynthesis exhibits additional layers of complexity compared with that of other organelles because they contain their own genome and dedicated ribosomes. Maintaining these components requires gene expression to be coordinated between the nucleo-cytoplasmic compartment and mitochondria in order to monitor organelle homeostasis and to integrate the responses to the physiological and developmental demands of the cell. Surprisingly, the parameters that are used to monitor or count mitochondrial abundance are not known, nor are the signalling pathways. Inhibiting the translation on mito-ribosomes genetically or with antibiotics can impair cell proliferation and has been attributed to defects in aerobic energy metabolism, even though proliferating cells rely primarily on glycolysis to fuel their metabolic demands. However, a recent study indicates that mitochondrial translational stress and the rescue mechanisms that relieve this stress cause the defect in cell proliferation and occur before any impairment of oxidative phosphorylation. Therefore, the process of mitochondrial translation in itself appears to be an important checkpoint for the monitoring of mitochondrial homeostasis and might have a role in establishing mitochondrial abundance within a cell. This hypothesis article will explore the evidence supporting a role for mito-ribosomes and translation in a mitochondria-counting mechanism.

  10. Lymphatic vessel development: fluid flow and valve-forming cells.

    PubMed

    Kume, Tsutomu

    2015-08-03

    Hemodynamic forces regulate many aspects of blood vessel disease and development, including susceptibility to atherosclerosis and remodeling of primary blood vessels into a mature vascular network. Vessels of the lymphatic circulatory system are also subjected to fluid flow-associated forces, but the molecular and cellular mechanisms by which these forces regulate the formation and maintenance of lymphatic vessels remain largely uncharacterized. This issue of the JCI includes two articles that begin to address how fluid flow influences lymphatic vessel development and function. Sweet et al. demonstrate that lymph flow is essential for the remodeling of primary lymphatic vessels, for ensuring the proper distribution of smooth muscle cells (SMCs), and for the development and maturation of lymphatic valves. Kazenwadel et al. show that flow-induced lymphatic valve development is initiated by the upregulation of GATA2, which has been linked to lymphedema in patients with Emberger syndrome. Together, these observations and future studies inspired by these results have potential to lead to the development of strategies for the treatment of lymphatic disorders.

  11. Assumed white blood cell count of 8,000 cells/μL overestimates malaria parasite density in the Brazilian Amazon.

    PubMed

    Alves-Junior, Eduardo R; Gomes, Luciano T; Ribatski-Silva, Daniele; Mendes, Clebson Rodrigues J; Leal-Santos, Fabio A; Simões, Luciano R; Mello, Marcia Beatriz C; Fontes, Cor Jesus F

    2014-01-01

    Quantification of parasite density is an important component in the diagnosis of malaria infection. The accuracy of this estimation varies according to the method used. The aim of this study was to assess the agreement between the parasite density values obtained with the assumed value of 8,000 cells/μL and the automated WBC count. Moreover, the same comparative analysis was carried out for other assumed values of WBCs. The study was carried out in Brazil with 403 malaria patients who were infected in different endemic areas of the Brazilian Amazon. The use of a fixed WBC count of 8,000 cells/μL to quantify parasite density in malaria patients led to overestimated parasitemia and resulted in low reliability when compared to the automated WBC count. Assumed values ranging between 5,000 and 6,000 cells/μL, and 5,500 cells/μL in particular, showed higher reliability and more similar values of parasite density when compared between the 2 methods. The findings show that assumed WBC count of 5,500 cells/μL could lead to a more accurate estimation of parasite density for malaria patients in this endemic region.

  12. Interleukin-2 Therapy Induces CD4 Downregulation, Which Decreases Circulating CD4 T Cell Counts, in African Green Monkeys

    PubMed Central

    Mudd, Joseph C.; Perkins, Molly R.; DiNapoli, Sarah R.; Hirsch, Vanessa M.

    2016-01-01

    ABSTRACT African green monkeys (AGMs) are natural hosts of simian immunodeficiency virus (SIVAGM). Because these animals do not develop simian AIDS despite maintaining high viral loads, there is considerable interest in determining how these animals have evolved to avoid SIV disease progression. Unlike nonnatural hosts of SIV, adult AGMs maintain low levels of CD4+ T cells at steady states and also have a large population of virus-resistant CD8αα T cells that lack CD4 expression despite maintaining T helper cell functionalities. In recent work, we have shown that homeostatic cytokines can induce CD4 downregulation in AGM T cells in vitro. Through administering therapeutic doses of recombinant human interleukin-2 (IL-2) to AGMs, we show here that this mechanism is operative in vivo. IL-2 therapy induced transient yet robust proliferation in all major T cell subsets. Within the CD4+ T cell population, those that were induced into cycle by IL-2 exhibited characteristics of CD4-to-CD8αα conversion. In all animals receiving IL-2, circulating CD4+ T cell counts and proportions tended to be lower and CD4− CD8αα+ T cell counts tended to be higher. Despite reductions in circulating target cells, the viral load was unaffected over the course of study. IMPORTANCE The data in this study identify that homeostatic cytokines can downregulate CD4 in vivo and, when given therapeutically, can induce AGMs to sustain very low levels of circulating CD4+ T cells without showing signs of immunodeficiency. PMID:27053558

  13. Effectiveness of magnetic fluid hyperthermia against Candida albicans cells.

    PubMed

    Chudzik, Barbara; Miaskowski, Arkadiusz; Surowiec, Zbigniew; Czernel, Grzegorz; Duluk, Tomasz; Marczuk, Andrzej; Gagoś, Mariusz

    2016-12-01

    Candida albicans is one of the most frequently isolated fungal pathogens causing opportunistic infections in humans. Targeted magnetic fluid hyperthermia (MFH) is a promising method in thermal therapy facilitating selective heating of pathogen cells like C. albicans. In the paper, we used meso-2,3-dimercaptosuccinic acid (DMSA)-coated magnetic nanoparticles (MNPs) and functionalised anti-C. albicans immunomagnetic nanoparticles (IMNPs) to investigate the potential of MFH in combating C. albicans cells in vitro. Using Mössbauer spectroscopy it was found that synthesised MNPs exhibited superparamagnetic phenomena. On the basis of calorimetric experiments, the maximum SAR (specific absorption rate) was found and a proper concentration of MNPs was established to control the temperature. MFH based on both DMSA-coated MNPs and functionalised anti-C. albicans IMNPs was more effective in combating C. albicans cells in vitro than thermostat hyperthermia. Especially promising results were obtained using functionalised IMNPs, which eradicated most of the pathogen colonies at the temperature of 43 °C.

  14. 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/μL 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/μL, 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 (β = 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/μL. Conclusion. Antenatal provision of cotrimoxazole for HIV-infected pregnant women with low CD4 cell counts may have indirect benefits for neonatal health. PMID:17083035

  15. Evaluation of nucleated red blood cell count by Sysmex XE-2100 in patients with thalassaemia or sickle cell anaemia and in neonates

    PubMed Central

    Buoro, Sabrina; Vavassori, Mauro; Pipitone, Silvia; Benegiamo, Anna; Lochis, Eleonora; Fumagalli, Sabina; Falanga, Anna; Marchetti, Marina; Crippa, Alberto; Ottomano, Cosimo; Lippi, Giuseppe

    2015-01-01

    Background Current haematology analysers have variable sensitivity and accuracy for counting nucleated red blood cells in samples with low values and in all those conditions characterised by altered sensitivity of red blood cells to the lysing process, such as in beta-thalassaemia or sickle-cell diseases and in neonates. The aim of our study was to evaluate the performance of the automated analyser XE-2100 at counting nucleated red blood cells in the above-mentioned three categories of subjects with potentially altered red blood cell lysis sensitivity and yet a need for accurate nucleated red blood cell counts. Materials and methods We measured nucleated red blood cell count by XE-2100 in peripheral blood samples of 187 subjects comprising 55 patients with beta-thalassaemia (40 major and 15 traits), 26 sickle-cell patients, 56 neonates and 50 normal subject. Results were compared with those obtained by optical microscopy. Agreement between average values of the two methods was estimated by means of Pearson’s correlation and bias analysis, whereas diagnostic accuracy was estimated by analysis of receiver operating characteristic curves. Results The comparison between the two methods showed a Pearson’s correlation of 0.99 (95% CI; 0.98–0.99; p<0.001) and bias of −0.61 (95% CI, −1.5–0.3). The area under the curve of the nucleated red blood cell count in all samples was 0.98 (95% CI, 0.96–1.00; p<0.001). Sub-analysis revealed an area under curve of 0.99 (95% CI, 0.98–1.00; p<0.001) for patients with thalassaemia, 0.94 (95% CI, 0.85–1.00; p<0.001) for patients with sickle cell anaemia, and 1.00 (95% CI, 1.0–1.0) for neonates. Discussion XE-2100 has excellent performance for nucleated red blood cell counting, especially in critical populations such as patients with haemoglobinopathies and neonates. PMID:25761322

  16. Stem cells from fetal membranes and amniotic fluid: markers for cell isolation and therapy.

    PubMed

    Pozzobon, Michela; Piccoli, Martina; De Coppi, Paolo

    2014-06-01

    Stem cell therapy is in constant need of new cell sources to conceive regenerative medicine approaches for diseases that are still without therapy. Scientists drew the attention toward amniotic membrane and amniotic fluid stem cells, since these sources possess many advantages: first of all as cells can be extracted from discarded foetal material it is inexpensive, secondly abundant stem cells can be obtained and finally, these stem cell sources are free from ethical considerations. Many studies have demonstrated the differentiation potential in vitro and in vivo toward mesenchymal and non-mesenchymal cell types; in addition the immune-modulatory properties make these cells a good candidate for allo- and xenotransplantation. This review offers an overview on markers characterisation and on the latest findings in pre-clinical or clinical setting of the stem cell populations isolated from these sources.

  17. Flow-cytometric total bacterial cell counts as a descriptive microbiological parameter for drinking water treatment processes.

    PubMed

    Hammes, Frederik; Berney, Michael; Wang, Yingying; Vital, Marius; Köster, Oliver; Egli, Thomas

    2008-01-01

    There are significantly more microbial cells in drinking water than what can be cultured on synthetic growth media. Nonetheless, cultivation-based heterotrophic plate counts (HPCs) are used worldwide as a general microbial quality parameter in drinking water treatment and distribution. Total bacterial cell concentrations are normally not considered during drinking water treatment as a design, operative or legislative parameters. This is mainly because easy and rapid methods for quantification of total bacterial cell concentrations have, up to now, not been available. As a consequence, the existing lack of data does not allow demonstrating the practical value of this parameter. In this study, we have used fluorescence staining of microbial cells with the nucleic acid stain SYBR((R)) Green I together with quantitative flow cytometry (FCM) to analyse total cell concentrations in water samples from a drinking water pilot plant. The plant treats surface water (Lake Zürich) through sequential ozonation, granular active carbon (GAC) filtration and membrane ultrafiltration (UF). The data were compared with adenosine tri-phosphate (ATP) measurements and conventional HPCs performed on the same water samples. We demonstrated that the impact of all three major treatment steps on the microbiology in the system could accurately be described with total cell counting: (1) ozonation caused chemical destruction of the bacterial cells; (2) GAC filtration facilitated significant regrowth of the microbial community; and (3) membrane UF physically removed the bacterial cells from the water. FCM typically detected 1-2 log units more than HPC, while ATP measurements were prone to interference from extracellular ATP released during the ozonation step in the treatment train. We have shown that total cell concentration measured with FCM is a rapid, easy, sensitive and importantly, a descriptive parameter of several widely applied drinking water treatment processes.

  18. Fluid Inclusion Gas Analysis

    DOE Data Explorer

    Dilley, Lorie

    2013-01-01

    Fluid inclusion gas analysis for wells in various geothermal areas. Analyses used in developing fluid inclusion stratigraphy for wells and defining fluids across the geothermal fields. Each sample has mass spectrum counts for 180 chemical species.

  19. The VIMOS Public Extragalactic Redshift Survey (VIPERS). On the recovery of the count-in-cell probability distribution function

    NASA Astrophysics Data System (ADS)

    Bel, J.; Branchini, E.; Di Porto, C.; Cucciati, O.; Granett, B. R.; Iovino, A.; de la Torre, S.; Marinoni, C.; Guzzo, L.; Moscardini, L.; Cappi, A.; Abbas, U.; Adami, C.; Arnouts, S.; Bolzonella, M.; Bottini, D.; Coupon, J.; Davidzon, I.; De Lucia, G.; Fritz, A.; Franzetti, P.; Fumana, M.; Garilli, B.; Ilbert, O.; Krywult, J.; Le Brun, V.; Le Fèvre, O.; Maccagni, D.; Małek, K.; Marulli, F.; McCracken, H. J.; Paioro, L.; Polletta, M.; Pollo, A.; Schlagenhaufer, H.; Scodeggio, M.; Tasca, L. A. M.; Tojeiro, R.; Vergani, D.; Zanichelli, A.; Burden, A.; Marchetti, A.; Mellier, Y.; Nichol, R. C.; Peacock, J. A.; Percival, W. J.; Phleps, S.; Wolk, M.

    2016-04-01

    We compare three methods to measure the count-in-cell probability density function of galaxies in a spectroscopic redshift survey. From this comparison we found that, when the sampling is low (the average number of object per cell is around unity), it is necessary to use a parametric method to model the galaxy distribution. We used a set of mock catalogues of VIPERS to verify if we were able to reconstruct the cell-count probability distribution once the observational strategy is applied. We find that, in the simulated catalogues, the probability distribution of galaxies is better represented by a Gamma expansion than a skewed log-normal distribution. Finally, we correct the cell-count probability distribution function from the angular selection effect of the VIMOS instrument and study the redshift and absolute magnitude dependency of the underlying galaxy density function in VIPERS from redshift 0.5 to 1.1. We found a very weak evolution of the probability density distribution function and that it is well approximated by a Gamma distribution, independently of the chosen tracers. Based on observations collected at the European Southern Observatory, Cerro Paranal, Chile, using the Very Large Telescope under programmes 182.A-0886 and partly 070.A-9007. Also based on observations obtained with MegaPrime/MegaCam, a joint project of CFHT and CEA/DAPNIA, at the Canada-France-Hawaii Telescope (CFHT), which is operated by the National Research Council (NRC) of Canada, the Institut National des Sciences de l'Univers of the Centre National de la Recherche Scientifique (CNRS) of France, and the University of Hawaii. This work is based in part on data products produced at TERAPIX and the Canadian Astronomy Data Centre as part of the Canada-France-Hawaii Telescope Legacy Survey, a collaborative project of NRC and CNRS. The VIPERS web site is http://www.vipers.inaf.it/

  20. Compressible cell gas models for asymmetric fluid criticality

    NASA Astrophysics Data System (ADS)

    Cerdeiriña, Claudio A.; Orkoulas, Gerassimos

    2017-03-01

    We thoroughly describe a class of models recently presented by Fisher and coworkers [Phys. Rev. Lett. 116, 040601 (2016)], 10.1103/PhysRevLett.116.040601. The crucial feature of such models, termed compressible cell gases (CCGs), is that the individual cell volumes of a lattice gas are allowed to fluctuate. They are studied via the seldom-used (μ , p , T ) ensemble, which leads to their exact mapping onto the Ising model. Remarkably, CCGs obey complete scaling, a formulation for the thermodynamic behavior of fluids near the gas-liquid critical point that accommodates features inherent to the asymmetric nature of this phase transition like the Yang-Yang (YY) and singular coexistence-curve diameter anomalies. The CCG0 models generated when volumes vary freely reveal local free volume fluctuations as the origin of these phenomena. Local energy-volume coupling is found to be another relevant microscopic factor. Furthermore, the CCG class is greatly extended by using the decoration transformation, with an interesting example being the Sastry-Debenedetti-Sciortino-Stanley model for hydrogen bonding in low-temperature water. The magnitude of anomalies is characterized by a single parameter, the YY ratio, which for the models so far considered here ranges from -∞ to 1/2 .

  1. Role of cells in freezing-induced cell-fluid-matrix interactions within engineered tissues.

    PubMed

    Seawright, Angela; Ozcelikkale, Altug; Dutton, Craig; Han, Bumsoo

    2013-09-01

    During cryopreservation, ice forms in the extracellular space resulting in freezing-induced deformation of the tissue, which can be detrimental to the extracellular matrix (ECM) microstructure. Meanwhile, cells dehydrate through an osmotically driven process as the intracellular water is transported to the extracellular space, increasing the volume of fluid for freezing. Therefore, this study examines the effects of cellular presence on tissue deformation and investigates the significance of intracellular water transport and cell-ECM interactions in freezing-induced cell-fluid-matrix interactions. Freezing-induced deformation characteristics were examined through cell image deformetry (CID) measurements of collagenous engineered tissues embedded with different concentrations of MCF7 breast cancer cells versus microspheres as their osmotically inactive counterparts. Additionally, the development of a biophysical model relates the freezing-induced expansion of the tissue due to the cellular water transport and the extracellular freezing thermodynamics for further verification. The magnitude of the freezing-induced dilatation was found to be not affected by the cellular water transport for the cell concentrations considered; however, the deformation patterns for different cell concentrations were different suggesting that cell-matrix interactions may have an effect. It was, therefore, determined that intracellular water transport during freezing was insignificant at the current experimental cell concentrations; however, it may be significant at concentrations similar to native tissue. Finally, the cell-matrix interactions provided mechanical support on the ECM to minimize the expansion regions in the tissues during freezing.

  2. Models to Estimate Lactation Curves of Milk Yield and Somatic Cell Count in Dairy Cows at the Herd Level for the Use in Simulations and Predictive Models

    PubMed Central

    Græsbøll, Kaare; Kirkeby, Carsten; Nielsen, Søren Saxmose; Halasa, Tariq; Toft, Nils; Christiansen, Lasse Engbo

    2016-01-01

    Typically, central milk recording data from dairy herds are recorded less than monthly. Over-fitting early in lactation periods is a challenge, which we explored in different ways by reducing the number of parameters needed to describe the milk yield and somatic cell count of individual cows. Furthermore, we investigated how the parameters of lactation models correlate between parities and from dam to offspring. The aim of the study was to provide simple and robust models for cow level milk yield and somatic cell count for fitting to sparse data to parameterize herd- and cow-specific simulation of dairy herds. Data from 610 Danish Holstein herds were used to determine parity traits in milk production regarding milk yield and somatic cell count of individual cows. Parity was stratified in first, second, and third and higher for milk, and first to sixth and higher for somatic cell count. Fitting of herd level parameters allowed for cow level lactation curves with three, two, or one parameters per lactation. Correlations of milk yield and somatic cell count were estimated between lactations and between dam and offspring. The shape of the lactation curves varied markedly between farms. The correlation between lactations for milk yield and somatic cell count was 0.2–0.6 and significant on more than 95% of farms. The variation in the daily milk yield was observed to be a source of variation to the somatic cell count, and the total somatic cell count was less correlated with the milk production than somatic cells per milliliter. A positive correlation was found between relative levels of the total somatic cell count and the milk yield. The variation of lactation and somatic cell count curves between farms highlights the importance of a herd level approach. The one-parameter per cow model using a herd level curve allows for estimating the cow production level from first the recording in the parity, while a two-parameter model requires more recordings for a credible

  3. Multiplicity Counting

    SciTech Connect

    Geist, William H.

    2015-12-01

    This set of slides begins by giving background and a review of neutron counting; three attributes of a verification item are discussed: 240Pueff mass; α, the ratio of (α,n) neutrons to spontaneous fission neutrons; and leakage multiplication. It then takes up neutron detector systems – theory & concepts (coincidence counting, moderation, die-away time); detector systems – some important details (deadtime, corrections); introduction to multiplicity counting; multiplicity electronics and example distributions; singles, doubles, and triples from measured multiplicity distributions; and the point model: multiplicity mathematics.

  4. Consecutive results of blood cell count and retrospective biodosimetry: useful tools of health protection regulation for radiation workers

    PubMed Central

    Jang, Seongjae; Lee, Jin Kyung; Cho, Minsu; Yang, Su San; Kim, Seung Hyun; Kim, Wan Tae

    2016-01-01

    Background Industrial radiography is known to be one of the most vulnerable lines of work among the range of different radiation work. According to the relevant law in Korea, every worker registered in this work should check their blood cell counts every year in addition to their thermoluminescent dosimeter (TLD) doses. Since the law was enacted, however, few follow-up studies have been carried out based on the obtained results. Objectives To ascertain the clinical usefulness of complete blood cell count (CBC) results and suggest a proper protocol of health protection for radiation workers. Methods After reviewing all the consecutive results of CBC and TLD doses from radiation workers registered nationwide, we selected two groups of high-risk radiation workers, CBC-high risk (CBC-HR) and TLD-high risk (TLD-HR) groups. A control group of unexposed healthy adults was also included. We compared the absorbed doses calculated by cytogenetic biodosimetry among those three groups, and examined possible confounding factors for each group. Results Both groups of high-risk radiation workers, CBC-HR and TLD-HR, showed higher chromosome aberrations than the control group. In the control group, previous medical history of a CT scan increased the frequency of chromosome aberrations. In contrast, the frequency of chromosome aberrations in the high-risk radiation workers was affected not by the previous CT history but only by the duration of their work. Conclusions We ascertain that reviewing consecutive results of blood cell counts and cytogenetic biodosimetry are useful complementary tools to TLD doses for health protection regulation. Several confounding factors including work duration and previous medical history need to be considered for the interpretation of biodosimetry results. PMID:27466611

  5. Genetic modification of primate amniotic fluid-derived stem cells produces pancreatic progenitor cells in vitro.

    PubMed

    Zhou, Yu; Mack, David L; Williams, J Koudy; Mirmalek-Sani, Sayed-Hadi; Moorefield, Emily; Chun, So-Young; Wang, Jun; Lorenzetti, Diego; Furth, Mark; Atala, Anthony; Soker, Shay

    2013-01-01

    Insulin therapy for type 1 diabetes does not prevent serious long-term complications including vascular disease, neuropathy, retinopathy and renal failure. Stem cells, including amniotic fluid-derived stem (AFS) cells - highly expansive, multipotent and nontumorigenic cells - could serve as an appropriate stem cell source for β-cell differentiation. In the current study we tested whether nonhuman primate (nhp)AFS cells ectopically expressing key pancreatic transcription factors were capable of differentiating into a β-cell-like cell phenotype in vitro. nhpAFS cells were obtained from Cynomolgus monkey amniotic fluid by immunomagnetic selection for a CD117 (c-kit)-positive population. RT-PCR for endodermal and pancreatic lineage-specific markers was performed on AFS cells after adenovirally transduced expression of PDX1, NGN3 and MAFA. Expression of MAFA was sufficient to induce insulin mRNA expression in nhpAFS cell lines, whereas a combination of MAFA, PDX1 and NGN3 further induced insulin expression, and also induced the expression of other important endocrine cell genes such as glucagon, NEUROD1, NKX2.2, ISL1 and PCSK2. Higher induction of these and other important pancreatic genes was achieved by growing the triply infected AFS cells in media supplemented with a combination of B27, betacellulin and nicotinamide, as well as culturing the cells on extracellular matrix-coated plates. The expression of pancreatic genes such as NEUROD1, glucagon and insulin progressively decreased with the decline of adenovirally expressed PDX1, NGN3 and MAFA. Together, these experiments suggest that forced expression of pancreatic transcription factors in primate AFS cells induces them towards the pancreatic lineage.

  6. OpenCFU, a new free and open-source software to count cell colonies and other circular objects.

    PubMed

    Geissmann, Quentin

    2013-01-01

    Counting circular objects such as cell colonies is an important source of information for biologists. Although this task is often time-consuming and subjective, it is still predominantly performed manually. The aim of the present work is to provide a new tool to enumerate circular objects from digital pictures and video streams. Here, I demonstrate that the created program, OpenCFU, is very robust, accurate and fast. In addition, it provides control over the processing parameters and is implemented in an intuitive and modern interface. OpenCFU is a cross-platform and open-source software freely available at http://opencfu.sourceforge.net.

  7. Can C-reactive protein and white blood cell count alone rule out an urgent condition in acute abdominal pain?

    PubMed

    Paolillo, Ciro; Spallino, Ilenia

    2016-02-01

    Up to 10% of all patients at the Emergency Department present for acute abdominal pain. The C-reactive protein (CRP) and white blood cell (WBC) are routinely determined as part of the workup of patients with abdominal pain. Three large prospective cohort studies comprising a total of 2961 adult patients with acute abdominal pain were selected. CRP levels and WBC counts were compared between patients with urgent and nonurgent final diagnoses. These studies conclude that the laboratory values individually are weak discriminators and cannot be used as a triage instrument in the selection of patients with acute abdominal pain requiring additional diagnostic tests.

  8. Cryptococcal meningitis manifesting as a large abdominal cyst in a HIV-infected patient with a CD4 count greater than 400 cells/mm(3).

    PubMed

    Crum-Cianflone, Nancy; Truett, April; R Wallace, Mark

    2008-05-01

    Cryptococcal meningitis usually occurs among HIV-positive patients with CD4 counts less than 100 cells/mm(3) and manifests as headaches, fevers, and mental status changes. We present an unusual case of cryptococcal meningitis in a 34-year-old HIV-positive man presenting as a large abdominal cyst at the ventriculoperitoneal shunt site despite receiving highly active antiretroviral therapy (HAART) for more than 5 years and having a CD4 count more than 400 cells/mm(3).

  9. Intestinal Parasitosis in Relation to Anti-Retroviral Therapy, CD4(+) T-cell Count and Diarrhea in HIV Patients.

    PubMed

    Khalil, Shehla; Mirdha, Bijay Ranjan; Sinha, Sanjeev; Panda, Ashutosh; Singh, Yogita; Joseph, Anju; Deb, Manorama

    2015-12-01

    Intestinal parasitic infections are one of the major causes of diarrhea in human immunodeficiency virus (HIV) seropositive individuals. Antiretroviral therapy has markedly reduced the incidence of many opportunistic infections, but parasite-related diarrhea still remains frequent and often underestimated especially in developing countries. The present hospital-based study was conducted to determine the spectrum of intestinal parasitosis in adult HIV/AIDS (acquired immunodeficiency syndrome) patients with or without diarrhea with the levels of CD4(+) T-cell counts. A total of 400 individuals were enrolled and were screened for intestinal parasitosis. Of these study population, 200 were HIV seropositives, and the remaining 200 were HIV uninfected individuals with or without diarrhea. Intestinal parasites were identified by using microscopy as well as PCR assay. A total of 130 (32.5%) out of 400 patients were positive for any kinds of intestinal parasites. The cumulative number of parasite positive patients was 152 due to multiple infections. A significant association of Cryptosporidium (P<0.001) was detected among individuals with CD4(+) T-cell counts less than 200 cells/μl.

  10. Selected complete blood cell count and plasma protein electrophoresis parameters in pet psittacine birds evaluated for illness.

    PubMed

    Briscoe, Jeleen A; Rosenthal, Karen L; Shofer, Frances S

    2010-06-01

    Veterinarians rely on results of both the complete blood cell count (CBC) and plasma protein electrophoresis (EPH) in conjunction with the results of the plasma biochemical analysis to evaluate the health status of avian patients. Because the CBC and protein EPH measure different aspects of the immune response to disease, both tests are recommended in avian patients to rule out infectious or inflammatory disease. To evaluate results of the CBC and protein EPH in pet psittacine birds, the records of 144 pet psittacine birds, comprising 11 genera, that were presented for suspected illness were reviewed. Results of the CBC (total white blood cell count and packed cell volume) and protein EPH (alpha, beta, and gamma globulin concentrations) from submitted blood samples from each bird were evaluated. Of the 144 birds, 63 (43.8%) had abnormal CBC results, and 25 (17.4%) had abnormal EPH measurements. Results of the CBC and protein EPH were within reference ranges in 73 birds (50.7%). Abnormal results of the CBC in conjunction with normal EPH results were present in 46 birds (31.9%), compared with 8 birds (5.6%) with normal results of the CBC and abnormal EPH results. The findings of this study could aid practitioners in evaluating psittacine patients and prioritizing the value of individual diagnostic tests.

  11. Intestinal Parasitosis in Relation to Anti-Retroviral Therapy, CD4+ T-cell Count and Diarrhea in HIV Patients

    PubMed Central

    Khalil, Shehla; Mirdha, Bijay Ranjan; Sinha, Sanjeev; Panda, Ashutosh; Singh, Yogita; Joseph, Anju; Deb, Manorama

    2015-01-01

    Intestinal parasitic infections are one of the major causes of diarrhea in human immunodeficiency virus (HIV) seropositive individuals. Antiretroviral therapy has markedly reduced the incidence of many opportunistic infections, but parasite-related diarrhea still remains frequent and often underestimated especially in developing countries. The present hospital-based study was conducted to determine the spectrum of intestinal parasitosis in adult HIV/AIDS (acquired immunodeficiency syndrome) patients with or without diarrhea with the levels of CD4+ T-cell counts. A total of 400 individuals were enrolled and were screened for intestinal parasitosis. Of these study population, 200 were HIV seropositives, and the remaining 200 were HIV uninfected individuals with or without diarrhea. Intestinal parasites were identified by using microscopy as well as PCR assay. A total of 130 (32.5%) out of 400 patients were positive for any kinds of intestinal parasites. The cumulative number of parasite positive patients was 152 due to multiple infections. A significant association of Cryptosporidium (P<0.001) was detected among individuals with CD4+ T-cell counts less than 200 cells/μl. PMID:26797437

  12. Susceptibility to cephalosporins of bacteria causing intramammary infections in dairy cows with a high somatic cell count in Germany.

    PubMed

    Wente, N; Zoche-Golob, V; Behr, M; Krömker, V

    2016-09-01

    The objective of this cross-sectional study was to determine the minimal inhibitory concentrations of cephalosporins of the first (cefalonium and cefapirin) and fourth generation (cefquinome) against bacteria isolated from intramammary infections in dairy cows with elevated somatic cell counts in Germany. Additionally, possible regional differences of the minimal inhibitory concentrations within Germany should be evaluated. In total, 6936 quarter milk samples from cows with a somatic cell count >200,000cells/ml were taken in 43 herds. The concentrations of the first generation cephalosporins inhibiting at least 90% of the isolates of a pathogen (MIC90) were ≥64μg/ml against Gram-negative bacteria and enterococci whereas the respective MIC90 against the other Gram-positive bacteria were ≤4μg/ml. The MIC90 of cefquinome were ≥16μg/ml against Gram-negative bacteria, bacilli and enterococci, and ≤2μg/ml against the other Gram-positive bacteria. Only the minimal inhibitory concentrations against coagulase-negative staphylococci differed significantly between regions in parametric survival models with shared frailties for the herds. However, the minimal inhibitory concentrations of cefquinome against staphylococci were higher than the minimal inhibitory concentrations of the tested cephalosporins of the first generation. Therefore, cefquinome should not be the first choice to treat staphylococcal mastitis in dairy cows.

  13. 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

  14. On-Orbit, Immuno-Based, Label-Free White Blood Cell Counting System with Microelectromechanical Sensor Technology (OILWBCS-MEMS)

    NASA Technical Reports Server (NTRS)

    Edmonds, Jessica

    2015-01-01

    Aurora Flight Sciences, in partnership with Draper Laboratory, has developed a miniaturized system to count white blood cells in microgravity environments. The system uses MEMS technology to simultaneously count total white blood cells, the five white blood cell differential subgroups, and various lymphocyte subtypes. The OILWBCS-MEMS detection technology works by immobilizing an array of white blood cell-specific antibodies on small, gold-coated membranes. When blood flows across the membranes, specific cells' surface protein antigens bind to their corresponding antibodies. This binding can be measured and correlated to cell counts. In Phase I, the partners demonstrated surface chemistry sensitivity and specificity for total white blood cells and two lymphocyte subtypes. In Phase II, a functional prototype demonstrated end-to-end operation. This rugged, miniaturized device requires minimal blood sample preparation and will be useful for both space flight and terrestrial applications.

  15. Tower counts

    USGS Publications Warehouse

    Woody, Carol Ann; Johnson, D.H.; Shrier, Brianna M.; O'Neal, Jennifer S.; Knutzen, John A.; Augerot, Xanthippe; O'Neal, Thomas A.; Pearsons, Todd N.

    2007-01-01

    Counting towers provide an accurate, low-cost, low-maintenance, low-technology, and easily mobilized escapement estimation program compared to other methods (e.g., weirs, hydroacoustics, mark-recapture, and aerial surveys) (Thompson 1962; Siebel 1967; Cousens et al. 1982; Symons and Waldichuk 1984; Anderson 2000; Alaska Department of Fish and Game 2003). Counting tower data has been found to be consistent with that of digital video counts (Edwards 2005). Counting towers do not interfere with natural fish migration patterns, nor are fish handled or stressed; however, their use is generally limited to clear rivers that meet specific site selection criteria. The data provided by counting tower sampling allow fishery managers to determine reproductive population size, estimate total return (escapement + catch) and its uncertainty, evaluate population productivity and trends, set harvest rates, determine spawning escapement goals, and forecast future returns (Alaska Department of Fish and Game 1974-2000 and 1975-2004). The number of spawning fish is determined by subtracting subsistence, sport-caught fish, and prespawn mortality from the total estimated escapement. The methods outlined in this protocol for tower counts can be used to provide reasonable estimates ( plus or minus 6%-10%) of reproductive salmon population size and run timing in clear rivers. 

  16. [A test for sperm cell survival in peritoneal fluid].

    PubMed

    Radwan, J; Niwald, W; Bielak, A; Pawlicki, J; Banaszczyk, R; Makuła, D

    1995-06-01

    The role of the peritoneal fluid in the physiology of reproduction, as well as in the transportation and survival of gametes, is little known. The authors have examined interactions between spermatozoa and the peritoneal fluid, collected during laparoscopy in the, so-called, survival test, from 42 infertile couples. The studied survival of spermatozoa in the peritoneal fluid was relatively high--19% after 48 hours--longer than in Menezo B2 fluid. Values of the test have been indicated, especially in cases of endometriosis-caused and idiopathic infertility.

  17. Amniotic Fluid Cells Are More Efficiently Reprogrammed to Pluripotency Than Adult Cells

    PubMed Central

    Galende, Elisa; Karakikes, Ioannis; Edelmann, Lisa; Desnick, Robert J.; Kerenyi, Thomas; Khoueiry, Georges; Lafferty, James; McGinn, Joseph T.; Brodman, Michael; Fuster, Valentin; Hajjar, Roger J.

    2010-01-01

    Abstract Recently, cultured human adult skin cells were reprogrammed to induced pluripotent stem (iPS) cells, which have characteristics similar to human embryonic stem (hES) cells. Patient-derived iPS cells offer genetic and immunologic advantages for cell and tissue replacement or engineering. The efficiency of generating human iPS cells has been very low; therefore an easily and efficiently reprogrammed cell type is highly desired. Here, we demonstrate that terminally differentiated human amniotic fluid (AF) skin cells provide an accessible source for efficiently generating abundant-induced pluripotent stem (AF-iPS) cells. By induction of pluripotency with the transcription factor quartet (OCT3/4, SOX2, KLF4, and c-MYC) the terminally differentiated, cultured AF skin cells formed iPS colonies approximately twice as fast and yielded nearly a two-hundred percent increase in number, compared to cultured adult skin cells. AF-iPS cells were identical to hES cells for morphological and growth characteristics, antigenic stem cell markers, stem cell gene expression, telomerase activity, in vitro and in vivo differentiation into the three germ layers and for their capacity to form embryoid bodies (EBs) and teratomas. Our findings provide a biological interesting conclusion that these fetal AF cells are more rapidly, easily, and efficiently reprogrammed to pluripotency than neonatal and adult cells. AF-iPS cells may have a “young,” more embryonic like epigenetic background, which may facilitate and accelerate pluripotency. The ability to efficiently and rapidly reprogram terminally differentiated AF skin cells and generate induced pluripotent stem cells provides an abundant iPS cell source for various basic studies and a potential for future patient-specific personalized therapies. PMID:20677926

  18. Flow Cytometry Approach to Quantify the Viability of Milk Somatic Cell Counts after Various Physico-Chemical Treatments

    PubMed Central

    Li, Na; Richoux, Romain; Perruchot, Marie-Hélène; Boutinaud, Marion; Mayol, Jean-François; Gagnaire, Valérie

    2015-01-01

    Flow cytometry has been used as a routine method to count somatic cells in milk, and to ascertain udder health and milk quality. However, few studies investigate the viability of somatic cells and even fewer at a subpopulation level to follow up how the cells can resist to various stresses that can be encountered during technological processes. To address this issue, a flow cytometry approach was used to simultaneously identify cell types of bovine milk using cell-specific antibodies and to measure the cell viability among the identified subpopulations by using a live/dead cell viability kit. Confirmation of the cell viability was performed by using conventional microscopy. Different physico-chemical treatments were carried out on standardized cell samples, such as heat treatment, various centrifugation rates and storage in milk or in PBS pH 7.4 for three days. Cytometry gating strategy was developed by using blood cell samples stored at 4°C in PBS and milk cell samples heat-treated at 80°C for 30 min as a control for the maximum (95.9%) and minimum (0.7%) values of cell viability respectively. Cell viability in the initial samples was 39.5% for all cells and varied for each cell population from 26.7% for PMNs, to 32.6% for macrophages, and 58.3% for lymphocytes. Regarding the physico-chemical treatments applied, somatic cells did not sustain heat treatment at 60°C and 80°C in contrast to changes in centrifugation rates, for which only the higher level, i.e. 5000×g led to a cell viability decrease, down to 9.4%, but no significant changes within the cell subpopulation distribution were observed. Finally, the somatic cells were better preserved in milk after 72h storage, in particular PMNs, that maintained a viability of 34.0 ± 2.9% compared to 4.9±1.9% in PBS, while there was almost no changes for macrophages (41.7 ± 5.7% in milk vs 31.2 ± 2.4% in PBS) and lymphocytes (25.3 ± 3.0% in milk vs 11.4 ± 3.1% in PBS). This study provides a new array to better

  19. Satellited 4q identified in amniotic fluid cells

    SciTech Connect

    Miller, I.; Hsieh, C.L.; Songster, G.

    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.

  20. [Cells in the cerebrospinal fluid of dogs and cats. Part 2].

    PubMed

    Grevel, V; Machus, B

    1992-02-01

    Three groups of cerebrospinal fluid (CSF) cells can be formed: 1. cells of the normal CSF, such as monocytes, small lymphocytes and occasionally cells of the ventricle system, 2. cells found in dogs and cats with neurologic disorders, such as reactive monocytes and lymphocytes, macrophages, neutrophils and eosinophils in addition to cells of the first group, 3. neoplastic cells. The different cells are introduced and their origin, function and occurrence are discussed. Mitotic figures, degenerated cells and artefacts are also mentioned.

  1. A new cell counting method to evaluate anti-tumor compound activity.

    PubMed

    Wang, Xue-Jian; Zhang, Xiu-Rong; Zhang, Lei; Li, Qing-Hua; Wang, Lin; Shi, Li-Hong; Fang, Chun-Yan

    2014-01-01

    Determining cell quantity is a common problem in cytology research and anti-tumor drug development. A simple and low-cost method was developed to determine monolayer and adherent-growth cell quantities. The cell nucleus is located in the cytoplasm, and is independent. Thus, the nucleus cannot make contact even if the cell density is heavy. This phenomenon is the foundation of accurate cell-nucleus recognition. The cell nucleus is easily recognizable in images after fluorescent staining because it is independent. A one-to-one relationship exists between the nucleus and the cell; therefore, this method can be used to determine the quantity of proliferating cells. Results indicated that the activity of the histone deacetylase inhibitor Z1 was effective after this method was used. The nude-mouse xenograft model also revealed the potent anti-tumor activity of Z1. This research presents a new anti-tumor-drug evaluation method.

  2. Spontaneous activity of cochlear hair cells triggered by fluid secretion mechanism in adjacent support cells

    PubMed Central

    Wang, Han Chin; Lin, Chun-Chieh; Cheung, Rocky; Zhang-Hooks, YingXin; Agarwal, Amit; Ellis-Davies, Graham; Rock, Jason; Bergles, Dwight E.

    2015-01-01

    Summary Spontaneous electrical activity of neurons in developing sensory systems promotes their maturation and proper connectivity. In the auditory system, spontaneous activity of cochlear inner hair cells (IHCs) is initiated by the release of ATP from glia-like inner supporting cells (ISCs), facilitating maturation of central pathways before hearing onset. Here, we find that ATP stimulates purinergic autoreceptors in ISCs, triggering Cl− efflux and osmotic cell shrinkage by opening TMEM16A Ca2+-activated Cl− channels. Release of Cl− from ISCs also forces K+ efflux, causing transient depolarization of IHCs near ATP release sites. Genetic deletion of TMEM16A markedly reduces the spontaneous activity of IHCs and spiral ganglion neurons in the developing cochlea, and prevents ATP-dependent shrinkage of supporting cells. These results indicate that support cells in the developing cochlea have adapted a pathway used for fluid secretion in other organs to induce periodic excitation of hair cells. PMID:26627734

  3. The search for true numbers of neurons and glial cells in the human brain: A review of 150 years of cell counting.

    PubMed

    von Bartheld, Christopher S; Bahney, Jami; Herculano-Houzel, Suzana

    2016-12-15

    For half a century, the human brain was believed to contain about 100 billion neurons and one trillion glial cells, with a glia:neuron ratio of 10:1. A new counting method, the isotropic fractionator, has challenged the notion that glia outnumber neurons and revived a question that was widely thought to have been resolved. The recently validated isotropic fractionator demonstrates a glia:neuron ratio of less than 1:1 and a total number of less than 100 billion glial cells in the human brain. A survey of original evidence shows that histological data always supported a 1:1 ratio of glia to neurons in the entire human brain, and a range of 40-130 billion glial cells. We review how the claim of one trillion glial cells originated, was perpetuated, and eventually refuted. We compile how numbers of neurons and glial cells in the adult human brain were reported and we examine the reasons for an erroneous consensus about the relative abundance of glial cells in human brains that persisted for half a century. Our review includes a brief history of cell counting in human brains, types of counting methods that were and are employed, ranges of previous estimates, and the current status of knowledge about the number of cells. We also discuss implications and consequences of the new insights into true numbers of glial cells in the human brain, and the promise and potential impact of the newly validated isotropic fractionator for reliable quantification of glia and neurons in neurological and psychiatric diseases. J. Comp. Neurol. 524:3865-3895, 2016. © 2016 Wiley Periodicals, Inc.

  4. How to count chromosomes in a cell: An overview of current and novel technologies.

    PubMed

    Bakker, Bjorn; van den Bos, Hilda; Lansdorp, Peter M; Foijer, Floris

    2015-05-01

    Aneuploidy, an aberrant number of chromosomes in a cell, is a feature of several syndromes associated with cognitive and developmental defects. In addition, aneuploidy is considered a hallmark of cancer cells and has been suggested to play a role in neurodegenerative disease. To better understand the relationship between aneuploidy and disease, various methods to measure the chromosome numbers in cells have been developed, each with their own advantages and limitations. While some methods rely on dividing cells and thus bias aneuploidy rates to that population, other, more unbiased methods can only detect the average aneuploidy rates in a cell population, cloaking cell-to-cell heterogeneity. Furthermore, some techniques are more prone to technical artefacts, which can result in over- or underestimation of aneuploidy rates. In this review, we provide an overview of several "traditional" karyotyping methods as well as the latest high throughput next generation sequencing karyotyping protocols with their respective advantages and disadvantages.

  5. Cell Imaging Counting as a Novel Ex Vivo Approach for Investigating Drug-Induced Hepatotoxicity in Zebrafish Larvae.

    PubMed

    Nguyen, Xuan-Bac; Kislyuk, Stanislav; Pham, Duc-Hung; Kecskés, Angela; Maes, Jan; Cabooter, Deirdre; Annaert, Pieter; De Witte, Peter; Ny, Annelii

    2017-02-08

    Drug-induced liver injury (DILI) is the most common reason for failures during the drug development process and for safety-related withdrawal of drugs from the pharmaceutical market. Therefore, having tools and techniques that can detect hepatotoxic properties in drug candidates at an early discovery stage is highly desirable. In this study, cell imaging counting was used to measure in a fast, straightforward, and unbiased way the effect of paracetamol and tetracycline, (compounds known to cause hepatotoxicity in humans) on the amount of DsRed-labeled hepatocytes recovered by protease digestion from Tg(fabp10a:DsRed) transgenic zebrafish. The outcome was in general comparable with the results obtained using two reference methods, i.e., visual analysis of liver morphology by fluorescence microscopy and size analysis of fluorescent 2D liver images. In addition, our study shows that administering compounds into the yolk is relevant in the framework of hepatotoxicity testing. Taken together, cell imaging counting provides a novel and rapid tool for screening hepatotoxicants in early stages of drug development. This method is also suitable for testing of other organ-related toxicities subject to the organs and tissues expressing fluorescent proteins in transgenic zebrafish lines.

  6. Connectivity and HIV-1 infection: role of CD4(+) T-cell counts and HIV-1 RNA copy number.

    PubMed

    Padierna-Olivos, L; Moreno-Altamirano, M M; Sánchez-Colón, S; Massó-Rojas, F; Sánchez-García, F J

    2000-12-01

    Following primary infection with human immunodeficiency virus (HIV)-1, antibodies against specific HIV-1 epitopes are elicited. However, non-HIV-1 specific antibodies, including autoantibodies, also arise. In fact, it has been proposed that such autoantibodies have an important role in the pathogenesis of HIV-1 infection. Because an imbalance in connectivity has been associated with autoimmune processes, we investigated the connectivity status of HIV-1-infected individuals. Moreover, we tested the possible role of viral load and CD4(+) T-cell counts, in connectivity, because these parameters appear to be important in the prognosis of HIV-1 infection. Results show that indeed, there is an alteration in connectivity in these patients, both for immunoglobulin (Ig)G and IgM, which is an immune alteration not previously identified in HIV-1 infection. In addition, our results show that viral load and CD4(+) T-cell counts are both equally important in defining the characteristic pattern of connectivity in HIV-1-infected individuals, and that neither is independently responsible for alterations in patient connectivity status.

  7. Cell Imaging Counting as a Novel Ex Vivo Approach for Investigating Drug-Induced Hepatotoxicity in Zebrafish Larvae

    PubMed Central

    Nguyen, Xuan-Bac; Kislyuk, Stanislav; Pham, Duc-Hung; Kecskés, Angela; Maes, Jan; Cabooter, Deirdre; Annaert, Pieter; De Witte, Peter; Ny, Annelii

    2017-01-01

    Drug-induced liver injury (DILI) is the most common reason for failures during the drug development process and for safety-related withdrawal of drugs from the pharmaceutical market. Therefore, having tools and techniques that can detect hepatotoxic properties in drug candidates at an early discovery stage is highly desirable. In this study, cell imaging counting was used to measure in a fast, straightforward, and unbiased way the effect of paracetamol and tetracycline, (compounds known to cause hepatotoxicity in humans) on the amount of DsRed-labeled hepatocytes recovered by protease digestion from Tg(fabp10a:DsRed) transgenic zebrafish. The outcome was in general comparable with the results obtained using two reference methods, i.e., visual analysis of liver morphology by fluorescence microscopy and size analysis of fluorescent 2D liver images. In addition, our study shows that administering compounds into the yolk is relevant in the framework of hepatotoxicity testing. Taken together, cell imaging counting provides a novel and rapid tool for screening hepatotoxicants in early stages of drug development. This method is also suitable for testing of other organ-related toxicities subject to the organs and tissues expressing fluorescent proteins in transgenic zebrafish lines. PMID:28208716

  8. Milk production, water consumption, and somatic cell count responses of cows subject to one to two volts of alternating current.

    PubMed

    Southwick, L H; Wilson, D J; Sears, P M

    1992-08-01

    A dairy farm located in central New York was visited because of complaints of electrical shock in the farmhouse shower and the milk house sink. As much as 2 volts AC of potential difference was measured between the waterline and the cow platform (cow-contact voltage). Voltage was coming from the primary neutral wire. The farm's electrical service was modified so that the farmstead could be connected or disconnected from the primary neutral wire at 2-week intervals for 12 weeks. When connected to the primary neutral wire, voltage between waterline and floor ranged between 0 and 1.8 volts, producing estimated current flow through cows of 3.6 to 4.9 mA; when disconnected from primary neutral wire, voltage between waterline and floor was less than 0.1 volt. There was no difference in mean milk production, bulk tank milk somatic cell count, or water consumption among periods when cows were exposed or unexposed to voltage. Despite statistical nonsignificance, the values for somatic cell count were lower and water consumption was higher when cows were exposed to voltage than when they were not.

  9. Short communication: effect of dry therapy using an intramammary infusion on bulk tank somatic cell count in sheep.

    PubMed

    Gonzalo, C; Linage, B; Carriedo, J A; Juárez, M T; Beneitez, E; Martínez, A; De La Fuente, L F

    2009-01-01

    A total of 3,141 records of bulk tank milk somatic cell counts (BTSCC) and bulk tank milk total bacterial counts (BTTBC) were obtained over 24 mo from 25 dairy flocks of Assaf ewes belonging to the Consortium for Ovine Promotion in Castilla-León, Spain, in which a complete dry therapy program was carried out in 10,313 ewes using an antibiotic infusion containing 100 mg of penethamate hydriodide, 280 mg of benethamine penicillin, and 100 mg of framycetin sulfate. The selection criteria for all flocks were BTSCC mean values > or =1,000 x 10(3) cells/mL and absence of dry therapy before the start of this experiment. Significant effects on log BTSCC were detected for treatment, milking system, flock within milking system, month within flock by treatment, the interactions treatment by milking system and flock by treatment within milking system, and log BTTBC. After dry therapy was implemented, log BTSCC decreased significantly in machine-milked flocks (5.95 +/- 0.007) compared with values before antibiotic treatment (6.13 +/- 0.008). The effect was observed at the beginning of the second lactation posttreatment (5.98 +/- 0.013). However, dry therapy was not effective in hand-milked flocks, suggesting poor hygiene conditions. A significant relationship was found between BTSCC and BTTBC; therefore, programs for improving milk hygiene should be implemented for both BTSCC and BTTBC variables at the same time.

  10. Interstitial fluid flow: simulation of mechanical environment of cells in the interosseous membrane

    NASA Astrophysics Data System (ADS)

    Yao, Wei; Ding, Guang-Hong

    2011-08-01

    In vitro experiments have shown that subtle fluid flow environment plays a significant role in living biological tissues, while there is no in vivo practical dynamical measurement of the interstitial fluid flow velocity. On the basis of a new finding that capillaries and collagen fibrils in the interosseous membrane form a parallel array, we set up a porous media model simulating the flow field with FLUENT software, studied the shear stress on interstitial cells' surface due to the interstitial fluid flow, and analyzed the effect of flow on protein space distribution around the cells. The numerical simulation results show that the parallel nature of capillaries could lead to directional interstitial fluid flow in the direction of capillaries. Interstitial fluid flow would induce shear stress on the membrane of interstitial cells, up to 30 Pa or so, which reaches or exceeds the threshold values of cells' biological response observed in vitro. Interstitial fluid flow would induce nonuniform spacial distribution of secretion protein of mast cells. Shear tress on cells could be affected by capillary parameters such as the distance between the adjacent capillaries, blood pressure and the permeability coefficient of capillary's wall. The interstitial pressure and the interstitial porosity could also affect the shear stress on cells. In conclusion, numerical simulation provides an effective way for in vivo dynamic interstitial velocity research, helps to set up the vivid subtle interstitial flow environment of cells, and is beneficial to understanding the physiological functions of interstitial fluid flow.

  11. Influence of Fluid Cell Design on the Frequency Response of AFM Microcantilevers in Liquid Media

    PubMed Central

    Motamedi, Ramin; Wood-Adams, Paula M.

    2008-01-01

    A study of the frequency response of AFM microcantilevers in liquid media contained in a commercial fluid cell is presented. Such systems exhibit complicated dynamics which are often not well described by available theories. Their dynamic behavior has a direct effect on the use of the AFM in dynamic mode while imaging in liquid or while extracting the rheological properties of the fluid. We explore the issues related to the design of the cantilever holder/fluid cell and propose an approach for evaluating, minimizing and recognizing the ultimate limitations of commercial cantilever holders. A technique for estimating the frequency response spectrum of the fluid cell itself from experimental data is presented. This spectrum can then be used to evaluate whether or not the fluid cell is suited for the desired purpose. PMID:27873849

  12. Dead cell counts during serum cultivation are underestimated by the fluorescent live/dead assay.

    PubMed

    Zhou, Shengda; Cui, Zhanfeng; Urban, Jill

    2011-05-01

    The live/dead fluorescent assay provides a quick method for assessing the proportion of live and dead cells in cell culture systems or tissues and is widely used. Dead cells are detected by the fluorescence produced when propidium iodide (PI) binds to DNA; PI and similar molecules are excluded from live cells but can penetrate dead cells because of their loss of membrane integrity. Here we investigated the effect of serum in the culture medium on the reliability of the method. We assessed viability of chondrocytes with/without serum using both a live/dead assay kit and also trypan blue staining. We found that after 2 days of culture, the DNA-binding dye PI could no longer detect dead cells if serum was present but they were readily detected in serum-free medium or if an inhibitor to DNase I was added to the serum-containing medium. Dead cells could be detected by trypan blue staining in all cultures. Hence dead cells are no longer detected as the DNase I present in serum degrades their DNA. DNA-binding dyes may thus not give a reliable estimate of the number of dead cells in systems that have been cultured in the presence of serum for several days.

  13. Estimated Costs for Delivery of HIV Antiretroviral Therapy to Individuals with CD4+ T-Cell Counts >350 cells/uL in Rural Uganda

    PubMed Central

    Jain, Vivek; Chang, Wei; Byonanebye, Dathan M.; Owaraganise, Asiphas; Twinomuhwezi, Ellon; Amanyire, Gideon; Black, Douglas; Marseille, Elliot; Kamya, Moses R.; Havlir, Diane V.; Kahn, James G.

    2015-01-01

    Background Evidence favoring earlier HIV ART initiation at high CD4+ T-cell counts (CD4>350/uL) has grown, and guidelines now recommend earlier HIV treatment. However, the cost of providing ART to individuals with CD4>350 in Sub-Saharan Africa has not been well estimated. This remains a major barrier to optimal global cost projections for accelerating the scale-up of ART. Our objective was to compute costs of ART delivery to high CD4+count individuals in a typical rural Ugandan health center-based HIV clinic, and use these data to construct scenarios of efficient ART scale-up. Methods Within a clinical study evaluating streamlined ART delivery to 197 individuals with CD4+ cell counts >350 cells/uL (EARLI Study: NCT01479634) in Mbarara, Uganda, we performed a micro-costing analysis of administrative records, ART prices, and time-and-motion analysis of staff work patterns. We computed observed per-person-per-year (ppy) costs, and constructed models estimating costs under several increasingly efficient ART scale-up scenarios using local salaries, lowest drug prices, optimized patient loads, and inclusion of viral load (VL) testing. Findings Among 197 individuals enrolled in the EARLI Study, median pre-ART CD4+ cell count was 569/uL (IQR 451–716). Observed ART delivery cost was $628 ppy at steady state. Models using local salaries and only core laboratory tests estimated costs of $529/$445 ppy (+/-VL testing, respectively). Models with lower salaries, lowest ART prices, and optimized healthcare worker schedules reduced costs by $100–200 ppy. Costs in a maximally efficient scale-up model were $320/$236 ppy (+/- VL testing). This included $39 for personnel, $106 for ART, $130/$46 for laboratory tests, and $46 for administrative/other costs. A key limitation of this study is its derivation and extrapolation of costs from one large rural treatment program of high CD4+ count individuals. Conclusions In a Ugandan HIV clinic, ART delivery costs—including VL testing

  14. 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 ...

  15. 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 ...

  16. 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 ...

  17. My oh my(osin): Insights into how auditory hair cells count, measure, and shape

    PubMed Central

    Pollock, Lana M.; Chou, Shih-Wei

    2016-01-01

    The mechanisms underlying mechanosensory hair bundle formation in auditory sensory cells are largely mysterious. In this issue, Lelli et al. (2016. J. Cell Biol. http://dx.doi.org/10.1083/jcb.201509017) reveal that a pair of molecular motors, myosin IIIa and myosin IIIb, is involved in the hair bundle’s morphology and hearing. PMID:26754648

  18. 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.

  19. HIV disease progression to CD4 count <200 cells/μL and death in Saskatoon, Saskatchewan

    PubMed Central

    Konrad, Stephanie; Skinner, Stuart; Kazadi, Germain Bukassa; Gartner, Kali; Lim, Hyun June

    2013-01-01

    OBJECTIVE: To characterize and identify determinants of HIV disease progression among a predominantly injection drug use (IDU) HIV population in the highly active antiretroviral therapy era. METHODS: The present retrospective study was based on 343 HIV patients diagnosed from 2005 to 2010 from two clinics in Saskatoon, Saskatchewan. Disease progression was defined as the time from diagnosis to immunological AIDS (CD4 count <200 cells/μL) and death. Uni- and multivariable Cox proportional hazards models were used. RESULTS: Of the 343 patients, 79% had a history of IDU, 77% were hepatitis C virus (HCV) coinfected and 67% were of Aboriginal descent. The one-year and three-year immunological AIDS-free probabilities were 78% and 53%, respectively. The one-year and three-year survival probabilities were 97% and 88%, respectively. Multicollinearity among IDU, HCV and ethnicity was observed and, thus, separate models were built. HCV coinfection (HR 2.9 [95% CI 1.2 to 6.9]) was a significant predictor of progression to immunological AIDS when controlling for baseline CD4 counts, treatment, age at diagnosis and year of diagnosis. For survival, only treatment use was a significant predictor (HR 0.34 [95% CI 0.1 to 0.8]). HCV coinfection was marginally significant (P=0.067). CONCLUSION: Baseline CD4 count, HCV coinfection, year of diagnosis and treatment use were significant predictors of disease progression. This highlights the importance of early treatment and the need for targeted interventions for these particularly vulnerable populations to slow disease progression. PMID:24421810

  20. 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).

  1. Testicular cytology indicates differences in Sertoli cell counts between "good freezer" and "poor freezer" bulls.

    PubMed

    Rajak, Shailendra Kumar; Thippeswamy, Vijetha Bajjalli; Kumaresan, Arumugam; Layek, Siddhartha Shankar; Mohanty, Tushar Kumar; Gaurav, Mukesh Kumar; Chakravarty, Atish Kumar; Datta, Tirtha Kumar; Manimaran, Ayyasamy; Prasad, Shiv

    2016-01-01

    In artificial insemination, poor quality of semen unsuitable for cryopreservation and susceptibility of spermatozoa to cryodamage in crossbred bulls have been a matter of concern. Present study was designed to identify the testicular cytology indices that might be used to predict the semen quality and cryotolerance of spermatozoa in bulls. Based on the ejaculate rejection rate and sperm cryotolerance, bulls (Holstein Friesian X Tharparkar crossbred) were classified into either good (producing good quality semen with spermatozoa having good cryotolerance; n = 4) or poor (producing poor quality semen with spermatozoa having poor cryotolerance; n = 4). Testicular cytology was studied in all the 8 bulls using fine needle aspiration technique. Testicular cytology of good bulls and poor bulls differed significantly. The proportion of Sertoli cells was significantly higher in good bulls (25.3 ± 1.6) compared to poor bulls (11.0 ± 0.8). The Sertoli cell index was 46.1 ± 5.0 in good bulls while it was only 13.8 ± 1.3 in poor bulls. The cut off values, as determined using Receiver Operating Characteristics analysis, indicate that the bulls having testicular cytogram comprising of < 15.5% Sertoli cells, < 24.3 Sertoli cell index and > 4.0 spermatogenic cells to Sertoli cell ratio might be a poor bull in terms of semen quality and cryotolerance of spermatozoa. The proportion of Sertoli cells in the testicular cytology had positive (P < 0.05) relationship with semen quality and cryotolerance of spermatozoa.

  2. Automatic Counting of Microglial Cells in Healthy and Glaucomatous Mouse Retinas

    PubMed Central

    Rojas, Blanca; Ramírez, Ana I.; de Hoz, Rosa; Salazar, Juan J.; Triviño, Alberto; Ramírez, José M.

    2015-01-01

    Proliferation of microglial cells has been considered a sign of glial activation and a hallmark of ongoing neurodegenerative diseases. Microglia activation is analyzed in animal models of different eye diseases. Numerous retinal samples are required for each of these studies to obtain relevant data of statistical significance. Because manual quantification of microglial cells is time consuming, the aim of this study was develop an algorithm for automatic identification of retinal microglia. Two groups of adult male Swiss mice were used: age-matched controls (naïve, n = 6) and mice subjected to unilateral laser-induced ocular hypertension (lasered; n = 9). In the latter group, both hypertensive eyes and contralateral untreated retinas were analyzed. Retinal whole mounts were immunostained with anti Iba-1 for detecting microglial cell populations. A new algorithm was developed in MATLAB for microglial quantification; it enabled the quantification of microglial cells in the inner and outer plexiform layers and evaluates the area of the retina occupied by Iba-1+ microglia in the nerve fiber-ganglion cell layer. The automatic method was applied to a set of 6,000 images. To validate the algorithm, mouse retinas were evaluated both manually and computationally; the program correctly assessed the number of cells (Pearson correlation R = 0.94 and R = 0.98 for the inner and outer plexiform layers respectively). Statistically significant differences in glial cell number were found between naïve, lasered eyes and contralateral eyes (P<0.05, naïve versus contralateral eyes; P<0.001, naïve versus lasered eyes and contralateral versus lasered eyes). The algorithm developed is a reliable and fast tool that can evaluate the number of microglial cells in naïve mouse retinas and in retinas exhibiting proliferation. The implementation of this new automatic method can enable faster quantification of microglial cells in retinal pathologies. PMID:26580208

  3. A dynamic pressure view cell for acoustic stimulation of fluids--Micro-bubble generation and fluid movement in porous media.

    PubMed

    Stewart, Robert A; Shaw, J M

    2015-09-01

    The development and baseline operation of an acoustic view cell for observing fluids, and fluid-fluid and fluid-solid interfaces in porous media over the frequency range of 10-5000 Hz is described. This range includes the industrially relevant frequency range 500-5000 Hz that is not covered by existing devices. Pressure waveforms of arbitrary shape are generated in a 17.46 mm ID by 200 mm and 690.5 mm long glass tubes at flow rates up to 200 ml/min using a syringe pump. Peak-to-peak amplitudes exceeding 80 kPa are readily realized at frequencies from 10 to 5000 Hz in bubble free fluids when actuated with 20 Vpp as exemplified using castor oil. At resonant frequencies, peak-to-peak pressure amplitudes exceeding 500 kPa were obtained (castor oil at 2100 Hz when actuated with 20 Vpp). Impacts of vibration on macroscopic liquid-liquid and liquid-vapour interfaces and interface movement are illustrated. Pressure wave transmission and attenuation in a fluid saturated porous medium, randomly packed 250-330 μm spherical silica beads, is also demonstrated. Attenuation differences and frequency shifts in resonant peaks are used to detect the presence and generation of dispersed micro-bubbles (<180 μm diameter), and bubbles within porous media that are not readily visualized. Envisioned applications include assessment of the impacts of vibration on reaction, mass transfer, and flow/flow pattern outcomes. This knowledge will inform laboratory and pilot scale process studies, where nuisance vibrations may affect the interpretation of process outcomes, and large scale or in situ processes in aquifers or hydrocarbon reservoirs where imposed vibration may be deployed to improve aspects of process performance. Future work will include miscible interface observation and quantitative measurements in the bulk and in porous media where the roles of micro-bubbles comprise subjects of special interest.

  4. Clinical features and outcome of T-lineage acute lymphoblastic leukemia in adults: a low initial white blood cell count, as well as a high count predict decreased survival rates.

    PubMed

    Yanada, Masamitsu; Jinnai, Itsuro; Takeuchi, Jin; Ueda, Takanori; Miyawaki, Shuichi; Tsuzuki, Motohiro; Hatta, Yoshihiro; Usui, Noriko; Wada, Hideho; Morii, Takeshi; Matsuda, Mitsuhiro; Kiyoi, Hitoshi; Okada, Masaya; Honda, Sumihisa; Miyazaki, Yasushi; Ohno, Ryuzo; Naoe, Tomoki

    2007-07-01

    Although biological and clinical features differ between B-lineage acute lymphoblastic leukemia (ALL) and T-lineage ALL (T-ALL), there have been few reports that focused on the prognosis for T-ALL in adults, primarily due to its rarity. Here, we studied the long-term outcomes and prognostic factors specific for adult T-ALL by combining patient data from the three prospective trials conducted by the Japan Adult Leukemia Study Group (JALSG). Among 559 patients whose immunophenotypes could be evaluated, 87 (15.6%) were identified as T-ALL. Of them, 66 patients (75.8%) achieved complete remission, and relapse occurred in 41 patients. With a median follow-up for surviving patients of 7.5 years, the probability of overall survival was 35.0% at 5 years. Risk factor analysis revealed that serum albumin levels, initial white blood cell (WBC) counts, and age had independent values for predicting survival. For WBC, not only the high-count group (50 x 10(9)l(-1) or higher), but also the low-count group (less than 3 x 10(9)l(-1)) showed a significantly lower survival rates than the intermediate-count group (p=0.0055 and 0.0037, respectively). Although our findings need confirmation, these results will be helpful in the identification of prognostically distinct subgroups within adult T-ALL.

  5. Evaluation of Baseline CD4+ T Cell Counts and ART Requirement in Newly Diagnosed HIV Seropositive Individuals in a Tertiary Care Hospital of Northern India

    PubMed Central

    Bhattar, Sonali; Mehra, Bhanu; Rawat, Deepti

    2016-01-01

    Introduction Antiretroviral Therapy (ART) has changed the outlook of Human Immune-deficiency Virus (HIV)/Acquired Immuno Deficiency Syndrome (AIDS) patients worldwide. Aim To analyse the trends in baseline CD4+ T cell counts and ART requirements in newly diagnosed HIV seropositive individuals in a Tertiary care hospital of Northern India. Materials and Methods Out of 1263 HIV seropositive clients identified from January 2012 to June 2014, the baseline CD4+ T cell counts of only those 470 clients were analysed, who registered at the linked ART centre. Results The mean baseline CD4+ count of the study group was 249.77±216.0cells/mm3 and that of male and female were 300.31±240.47cells/mm3 and 232.38±204.25cells/mm3 respectively. A total of 259 of 334 (77.54%) HIV reactive males, 83 of 130 (63.85%) HIV reactive females and overall 348 of 470 (74.04%) required antiretroviral treatment on enrolment. Conclusion In the present study, about three-fourth of newly diagnosed HIV positive Indian patients required initiation of ART at registration. The relatively low baseline CD4+ T cell counts in this population highlights the need for timely baseline CD4+ counts testing of HIV positive patients and the urgency of initiating treatment in HIV reactive individuals in Indian health care settings. PMID:28050367

  6. 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…

  7. 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)

  8. Effect of somatic cell count level on functional longevity in Valle del Belice dairy sheep assessed using survival analysis.

    PubMed

    Riggio, V; Maizon, D O; Portolano, B; Bovenhuis, H; van Arendonk, J A M

    2009-12-01

    The objectives of this study were to evaluate the effect of somatic cell count (SCC) on functional longevity and to estimate the heritability of functional longevity using survival analysis in Valle del Belice dairy sheep. A total of 4,880 lactations of 2,190 ewes from 11 flocks were used. In this study, SCC was considered as an indication of subclinical mastitis. In case of clinical cases, identified by the technicians at milking time, test-day weights and milk samples of those ewes were not considered. Somatic cells were analyzed as counts, without any transformation, and were grouped in 3 classes based on the observed SCC maximum (mxSCC). The mxSCC classes, expressed as 10(3) cells/mL, were classified as 1 if mxSCC or= 1,000. An increase in SCC was associated with an increased hazard of being culled. Ewes in the highest class of SCC on a test-day had a 20% higher hazard of being culled than those in the lowest class. Therefore, SCC played a role in culling decisions of Valle del Belice dairy sheep farmers. The heritability estimate for functional longevity was 7% on the logarithmic scale and 11% on the real scale, indicating that selection for this trait is possible in sheep. The flock-year-season effect explained 19% of the variation on the logarithmic scale and 27% of the variation on the real scale.

  9. 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

  10. Genetic parameters for faecal egg count, packed-cell volume and body-weight in Santa Inês lambs.

    PubMed

    Lôbo, Raimundo N B; Vieira, Luiz S; de Oliveira, Amaury A; Muniz, Evandro N; da Silva, José M

    2009-04-01

    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.

  11. Absolute lymphocyte count is associated with minimal residual disease level in childhood B-cell precursor acute lymphoblastic leukemia.

    PubMed

    Shen, Hong-Qiang; Feng, Jian-Hua; Tang, Yong-Min; Song, Hua; Yang, Shi-Long; Shi, Shu-Wen; Xu, Wei-Qun

    2013-06-01

    The prognostic value of absolute lymphocyte count (ALC) has been a recent matter of debate in childhood acute lymphoblastic leukemia (ALL). In the current study, ALCs at the time of diagnosis (ALC-0), after 7 days of initial therapy (ALC-8) and at interim of the induction therapy (ALC-22) were examined in Chinese children with B-cell precursor (BCP) ALL and correlated with the level of minimal residual disease (MRD) at day 22 of induction therapy. Medical and laboratory records of 140 patients diagnosed with childhood BCP ALL were retrieved and analyzed. ALC-22 is significantly correlated with MRD level at day 22 of therapy and can be a good prognostic factor for childhood BCP-ALL. Furthermore, lymphocyte count at initial diagnosis is correlated with MRD level at day 22 in childhood BCP-ALL with the immnunophenotype of CD19(pos)/CD10(pos)/CD34(pos)/CD45(neg) and role as a new prognostic factor was determined.

  12. 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

  13. Segmentation technique of complex image scene for an automatic blood-cell-counting system

    NASA Astrophysics Data System (ADS)

    Kovalev, Vassili A.; Grigoriev, Andrei Y.; Ahn, Hyo-Sok; Myshkin, Nickolai K.

    1996-04-01

    The paper presents a method for automatic localization and segmentation of white blood cells (WBCs) with color images to develop an efficient automated leukocyte counter by using pattern recognition-based slide readers. The segmentation techniques consist of the following steps. On the first a smear image acquired at the low magnification. The next is extraction of WBC nuclei by chromatic properties and image mapping. After this the cells clustered according to the distances between them and regions of interest (ROI) determined. Image of ROI captured at the high magnification and its validity checked. Then nucleus segments extracted and grouped into prospective cells. The detection of blood cells is based on the intensity of G image plane and the balance between G and B intensity of the nuclei. A cytoplasm region approximated by a circle area around the nucleus center. Finally, the cytoplasm area cleaned considering a priori knowledge of background color and possible cell occlusions. The result of the segmentation is presented in the form of a cell location list and image template in which every pixel is assigned to a label such as Background, Cytoplasm, Nucleus, Hole, etc. The proposed technique has yielded correct segmentation of complex image scenes for blood smears prepared by ordinary manual staining methods in 99% of tested images.

  14. Development of Microelectrode Arrays Using Electroless Plating for CMOS-Based Direct Counting of Bacterial and HeLa Cells.

    PubMed

    Niitsu, Kiichi; Ota, Shoko; Gamo, Kohei; Kondo, Hiroki; Hori, Masaru; Nakazato, Kazuo

    2015-10-01

    The development of two new types of high-density, electroless plated microelectrode arrays for CMOS-based high-sensitivity direct bacteria and HeLa cell counting are presented. For emerging high-sensitivity direct pathogen counting, two technical challenges must be addressed. One is the formation of a bacteria-sized microelectrode, and the other is the development of a high-sensitivity and high-speed amperometry circuit. The requirement for microelectrode formation is that the gold microelectrodes are required to be as small as the target cell. By improving a self-aligned electroless plating technique, the dimensions of the microelectrodes on a CMOS sensor chip in this work were successfully reduced to 1.2 μm × 2.05 μm. This is 1/20th of the smallest size reported in the literature. Since a bacteria-sized microelectrode has a severe limitation on the current flow, the amperometry circuit has to have a high sensitivity and high speed with low noise. In this work, a current buffer was inserted to mitigate the potential fluctuation. Three test chips were fabricated using a 0.6- μm CMOS process: two with 1.2 μm × 2.05 μm (1024 × 1024 and 4 × 4) sensor arrays and one with 6- μm square (16 × 16) sensor arrays; and the microelectrodes were formed on them using electroless plating. The uniformity among the 1024 × 1024 electrodes arranged with a pitch of 3.6 μm × 4.45 μm was optically verified. For improving sensitivity, the trenches on each microelectrode were developed and verified optically and electrochemically for the first time. Higher sensitivity can be achieved by introducing a trench structure than by using a conventional microelectrode formed by contact photolithography. Cyclic voltammetry (CV) measurements obtained using the 1.2 μm × 2.05 μm 4 × 4 and 6- μm square 16 × 16 sensor array with electroless-plated microelectrodes successfully demonstrated direct counting of the bacteria-sized microbeads and HeLa cells.

  15. B cell depletion in HIV-1 subtype A infected Ugandan adults: relationship to CD4 T cell count, viral load and humoral immune responses.

    PubMed

    Oballah, Peter; Flach, Britta; Eller, Leigh A; Eller, Michael A; Ouma, Benson; de Souza, Mark; Kibuuka, Hannah N; Wabwire-Mangen, Fred; Brown, Bruce K; Michael, Nelson L; Robb, Merlin L; Montefiori, David; Polonis, Victoria R

    2011-01-01

    To better understand the nature of B cell dysfunctions in subjects infected with HIV-1 subtype A, a rural cohort of 50 treatment-naïve Ugandan patients chronically infected with HIV-1 subtype A was studied, and the relationship between B cell depletion and HIV disease was assessed. B cell absolute counts were found to be significantly lower in HIV-1+ patients, when compared to community matched negative controls (p<0.0001). HIV-1-infected patients displayed variable functional and binding antibody titers that showed no correlation with viral load or CD4+ T cell count. However, B cell absolute counts were found to correlate inversely with neutralizing antibody (NAb) titers against subtype A (p = 0.05) and subtype CRF02_AG (p = 0.02) viruses. A positive correlation was observed between subtype A gp120 binding antibody titers and NAb breadth (p = 0.02) and mean titer against the 10 viruses (p = 0.0002). In addition, HIV-1 subtype A sera showed preferential neutralization of the 5 subtype A or CRF02_AG pseudoviruses, as compared with 5 pseudoviruses from subtypes B, C or D (p<0.001). These data demonstrate that in patients with chronic HIV-1 subtype A infection, significant B cell depletion can be observed, the degree of which does not appear to be associated with a decrease in functional antibodies. These findings also highlight the potential importance of subtype in the specificity of cross-clade neutralization in HIV-1 infection.

  16. Fluid shear stress sensitizes cancer cells to receptor-mediated apoptosis via trimeric death receptors

    NASA Astrophysics Data System (ADS)

    Mitchell, Michael J.; King, Michael R.

    2013-01-01

    Cancer metastasis, the process of cancer cell migration from a primary to distal location, typically leads to a poor patient prognosis. Hematogenous metastasis is initiated by intravasation of circulating tumor cells (CTCs) into the bloodstream, which are then believed to adhere to the luminal surface of the endothelium and extravasate into distal locations. Apoptotic agents such as tumor necrosis factor apoptosis-inducing ligand (TRAIL), whether in soluble ligand form or expressed on the surface of natural killer cells, have shown promise in treating CTCs to reduce the probability of metastasis. The role of hemodynamic shear forces in altering the cancer cell response to apoptotic agents has not been previously investigated. Here, we report that human colon cancer COLO 205 and prostate cancer PC-3 cells exposed to a uniform fluid shear stress in a cone-and-plate viscometer become sensitized to TRAIL-induced apoptosis. Shear-induced sensitization directly correlates with the application of fluid shear stress, and TRAIL-induced apoptosis increases in a fluid shear stress force- and time-dependent manner. In contrast, TRAIL-induced necrosis is not affected by the application fluid shear stress. Interestingly, fluid shear stress does not sensitize cancer cells to apoptosis when treated with doxorubicin, which also induces apoptosis in cancer cells. Caspase inhibition experiments reveal that shear stress-induced sensitization to TRAIL occurs via caspase-dependent apoptosis. These results suggest that physiological fluid shear forces can modulate receptor-mediated apoptosis of cancer cells in the presence of apoptotic agents.

  17. Influence of somatic cell counts and breed on physico-chemical and sensory characteristics of hard ewes'-milk cheeses.

    PubMed

    Revilla, Isabel; Lurueña-Martínez, Miguel A; Vivar-Quintana, Ana M

    2009-08-01

    The aim of the present work was to perform a physico-chemical, descriptive quantitative and consumer-preference analysis of hard ewes'-milk cheeses that had been matured for one year and to determine the correlations between the variables studied. The cheeses were elaborated with milk from three breeds of sheep (Castellana, Churra and Assaf) with different somatic cell counts (lower than 500,000 cells ml-1; between 1,000,000 and 1,500,000 cells ml-1, and more than 2,500,000 cells ml-1). The results show that the cheeses elaborated with milk with high SCC had lower values of dry extract and fat and high values of pH and fat acidity and were described as pungent, granulose and less creamy. Regarding the effect of breed, the cheeses made with milk from the Churra breed had lower values for fat and those made with Assaf breed milk were significantly more rancid. The study of correlations showed that creaminess was positively correlated with the dry extract and total fat content and negatively correlated with ash and fat acidity, indeed grainy texture and pungency had the opposite sign in their correlation with these latter variables. The yellow colour was positively correlated with ash and negatively with protein. Finally, the consumer preferences reveals that the less accepted cheeses showed the higher values for rancidness and pungency and they were less likely to accept the cheeses made with Assaf breed milk.

  18. The effects of storage temperature on goat milk somatic cell count using the DeLaval counter.

    PubMed

    Sanchez-Macias, Davinia; Castro, Noemi; Moreno-Indias, Isabel; Morales-delaNuez, Antonio; Briggs, Heather; Capote, Juan; Argüello, Anastasio

    2010-10-01

    This study investigated the influence of storage temperature and storage time on goat milk somatic cell counts (SCCs) determined using the DeLaval cell counter (DCC). SCCs were measured in 40 Majorera goat milk samples using the DCC device. Samples were grouped from high score (>2,750 x 10(3) cells/mL) to low score (<630 x 10(3) cell/mL) according to the SCC. Each milk sample was divided into four aliquots and stored at four different temperatures (4 degrees C, 21 degrees C, 36 degrees C or 45 degrees C). The SCC was recorded every hour for 12 hours. Storage of goat milk with a high SCC for 5, 5, 2 or 1 hour at 4 degrees C, 21 degrees C, 36 degrees C or 45 degrees C, respectively, decreased the SCC value compared to fresh milk. The goat milk SCC was lower after 1 hour of storage than that determined for fresh milk at any tested temperature in low-SCC samples. The data presented herein suggest that regardless of storage temperature, goat milk samples should not be stored for more than 1 hour before measurement of SCC with a DCC device.

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

    NASA Astrophysics Data System (ADS)

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

    2010-07-01

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

  20. B-cell targeted treatments for lupus: the journey counts as much as the destination.

    PubMed

    Falgarone, Géraldine; Dhote, Robin; Boissier, Marie-Christophe

    2012-10-01

    Obstacles facing therapeutic trials in systemic lupus erythematosus (SLE) include the low incidence, seriousness, complexity, and clinical polymorphism of the disease. A large-scale multicenter design has been required in most cases. Over the last few years, several biologics have been evaluated as treatments for lupus nephritis or for the skin and joint manifestations of SLE. The central role for the B-cell in SLE, together with improved knowledge of the targets on the B-cell surface, has prompted efforts to develop monoclonal antibodies as treatments for SLE. The two available monoclonal antibodies are rituximab (anti-CD20 antibody) and belimumab (anti-BlyS antibody). The results obtained with belimumab were used to develop a new measurement tool, the SLE Responder Index (SRI), and prompted an application for a license to use belimumab in SLE. Other targets identified on the B-cell surface are being evaluated.

  1. Effect of dissolved oxygen, temperature, initial cell count, and sugar concentration on the viability of Saccharomyces cerevisiae in rapid fermentations.

    PubMed

    Nagodawithana, T W; Castellano, C; Steinkraus, K H

    1974-09-01

    By using 7 x 10(8) cells of Saccharomyces cerevisiae per ml with which 25 degrees Brix honey solutions were fermented to 9.5% (wt/vol; 12% vol/vol) ethanol in 2.5 to 3 h at 30 C, i.e., rapid fermentation, the death rate was found to be high, with only 2.1% of the yeast cells surviving at the end of 3 h under anaerobic conditions. As the dissolved oxygen in the medium was increased from 0 to 13 to 20 to 100% in rapid fermentations at 30 C, there was a progressive increase in the percentage of cells surviving. The ethanol production rate and total were not seriously affected by a dissolved oxygen concentration of 13%, but fermentation was retarded by 20% dissolved oxygen and still further decreased as the dissolved oxygen content reached 100%. When the fermentation temperature was decreased to 15 C (at 13% dissolved oxygen), the rate of fermentation decreased, and the fermentation time to 9.5% ethanol (wt/vol) increased to 6 h. It was found that the higher the temperature between 15 and 30 C, the greater the rate of death as initial cell counts were increased from 1.1 x 10(7) to 7.8 x 10(8) cells per ml. At the lowest level of inoculum, 1.1 x 10(7) cells per ml, there was actual multiplication, even at 30 C; however, the fermentation was no longer rapid. The addition of 15% sugar, initially followed after an hour by the remaining 10%, or addition of the sugar in increments of 2.5 or 5% yielded a better survival rate of yeast cells than when the fermentation was initiated with 25% sugar.

  2. AMNIOTIC FLUID STEM CELLS: THE KNOWN, THE UNKNOWN AND POTENTIAL REGENERATIVE MEDICINE APPLICATIONS.

    PubMed

    Loukogeorgakis, Stavros P; De Coppi, Paolo

    2016-12-23

    The amniotic fluid has been identified as an untapped source of cells with broad potential, which possess immunomodulatory properties and don't have the ethical and legal limitations of embryonic stem cells. CD117(c-Kit)+ cells selected from amniotic fluid have been shown to differentiate into cell lineages representing all three embryonic germ layers without generating tumours, making them ideal candidates for regenerative medicine applications. Moreover, their ability to engraft in injured organs and modulate immune and repair responses of host tissues, suggest that transplantation of such cells may be useful for the treatment of various degenerative and inflammatory diseases. Although significant questions remain regarding the origin, heterogeneous phenotype and expansion potential of amniotic fluid stem cells, evidence to date supports their potential role as a valuable stem cell source for the field of regenerative medicine. This article is protected by copyright. All rights reserved.

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

    PubMed Central

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

    2013-01-01

    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

  4. Experimental colonization of the bovine teat duct with Corynebacterium bovis and the effect on milk somatic cell counts.

    PubMed Central

    Brooks, B W; Barnum, D A

    1984-01-01

    Colonization with Corynebacterium bovis was established in 59 of 64 (92%), 58 of 59 (98%) and 19 of 34 (56%) of uninfected bovine mammary quarters following inoculation of 83.3 X 10(4) colony-forming units (CFU) of the organism into the teat cistern, 4.7 X 10(3) CFU 5 mm into the teat duct or by exposure of the teat orifice to a milk culture containing 1.6 X 10(7) CFU/mL respectively. Mean somatic cell counts for foremilk samples from 122 quarters were significantly higher after colonization with C. bovis (145,900/mL) compared to before exposure (130,900/mL). PMID:6722643

  5. Total and differential white blood cell counts in Caiman latirostris after in ovo and in vivo exposure to insecticides.

    PubMed

    Latorre, María A; Romito, María L; Larriera, Alejandro; Poletta, Gisela L; Siroski, Pablo A

    2016-11-01

    Agricultural activities associated mainly with soybean crops affect the natural environment and wildlife by habitat destruction and the extensive use of agrochemicals. The aim of this study was to evaluate immunotoxic effects of the insecticides cypermethrin (CYP) and endosulfan (END) in Caiman latirostris analyzing total blood cell count (TWBC) and differential white blood cell count (DWBC) after in ovo and in vivo exposure. Eggs (in ovo) and hatchlings (in vivo) from nests harvested in natural habitats were artificially incubated and reared under controlled conditions in the Proyecto Yacaré (Gob.Santa Fe/MUPCN) facilities. Exposure of embryos was performed by topication on the eggshell during the first stage of development. The treatments were distilled water (negative control; NC), ethanol (vehicle control; VC), four groups treated with different concentrations of CYP and four groups with END. In vivo exposure was performed by immersion; treatments were NC, VC, two groups exposed to CYP and two to END. After embryonic exposure to the insecticides, no differences were found in TWBC or DWBC among the neonates exposed to pesticides versus controls. In the in vivo scenario, similar results were obtained for TWBC, but DWBC data showed differences between NC hatchlings and CYP-1 hosts for heterophil, lymphocyte and monocyte levels, and between NC and END-1 hosts for lymphocyte and monocyte levels. Research on the effects of pesticide exposure on this species is of special interest not only to assess the impact on caiman populations, but also to further characterize the species as a potential sentinel of ecosystem health.

  6. Inflammatory Cytokines and White Blood Cell Counts Response to Environmental Levels of Diesel Exhaust and Ozone Inhalation Exposures

    PubMed Central

    Stiegel, Matthew A.; Pleil, Joachim D.; Sobus, Jon R.; Madden, Michael C.

    2016-01-01

    Epidemiological observations of urban inhalation exposures to diesel exhaust (DE) and ozone (O3) have shown pre-clinical cardiopulmonary responses in humans. Identifying the key biological mechanisms that initiate these health bioindicators is difficult due to variability in environmental exposure in time and from person to person. Previously, environmentally controlled human exposure chambers have been used to study DE and O3 dose-response patterns separately, but investigation of co-exposures has not been performed under controlled conditions. Because a mixture is a more realistic exposure scenario for the general public, in this study we investigate the relationships of urban levels of urban-level DE exposure (300 μg/m3), O3 (0.3 ppm), DE + O3 co-exposure, and innate immune system responses. Fifteen healthy human volunteers were studied for changes in ten inflammatory cytokines (interleukins 1β, 2, 4, 5, 8, 10, 12p70 and 13, IFN-γ, and TNF-α) and counts of three white blood cell types (lymphocytes, monocytes, and neutrophils) following controlled exposures to DE, O3, and DE+O3. The results show subtle cytokines responses to the diesel-only and ozone-only exposures, and that a more complex (possibly synergistic) relationship exists in the combination of these two exposures with suppression of IL-5, IL-12p70, IFN-γ, and TNF-α that persists up to 22-hours for IFN-γ and TNF-α. The white blood cell differential counts showed significant monocyte and lymphocyte decreases and neutrophil increases following the DE + O3 exposure; lymphocytes and neutrophils changes also persist for at least 22-hours. Because human studies must be conducted under strict safety protocols at environmental levels, these effects are subtle and are generally only seen with detailed statistical analysis. This study indicates that the observed associations between environmental exposures and cardiopulmonary effects are possibly mediated by inflammatory response mechanisms. PMID:27058360

  7. Elevated white blood cell count is associated with higher risk of glucose metabolism disorders in middle-aged and elderly Chinese people.

    PubMed

    Jiang, Hua; Yan, Wen-Hua; Li, Chan-Juan; Wang, An-Ping; Dou, Jing-Tao; Mu, Yi-Ming

    2014-05-20

    White blood cell (WBC) count has been associated with diabetic risk, but whether the correlation is independent of other risk factors has hardly been studied. Moreover, very few such studies with large sample sizes have been conducted in Chinese. Therefore, we investigated the relationship between WBC count and glucose metabolism in China. We also examined the relevant variables of WBC count. A total of 9,697 subjects (mean age, 58.0 ± 9.1 years) were recruited. The subjects were classified into four groups, including subjects with normal glucose tolerance, isolated impaired fasting glucose, impaired glucose tolerance and type 2 diabetes mellitus (T2DM). We found that WBC count increased as glucose metabolism disorders exacerbated. WBC count was also positively correlated with waist hip ratio, body mass index, smoking, triglycerides, glycosylated haemoglobin A1c (HbA1c) and 2-h postprandial glucose. In addition, high density lipoprotein and the female gender were inversely correlated with WBC levels. In patients with previously diagnosed T2DM, the course of T2DM was not correlated with WBC count. Our findings indicate that elevated WBC count is independently associated with worsening of glucose metabolism in middle-aged and elderly Chinese. In addition, loss of weight, smoking cessation, lipid-modifying therapies, and control of postprandial plasma glucose and HbA1c may ameliorate the chronic low-grade inflammation.

  8. 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

  9. Effect of intramammary injection of rboGM-CSF on milk levels of chemiluminescence activity, somatic cell count, and Staphylococcus aureus count in Holstein cows with S. aureus subclinical mastitis

    PubMed Central

    2004-01-01

    Abstract The effect of intramammary injection of recombinant bovine granulocyte-macrophage colony-stimulating factor (rboGM-CSF, 400 μg/10 mL) on quarter milk levels of chemiluminescence (CL) activity, and somatic cell count (SCC) and shedding pattern of Staphylococcus aureus was investigated. Ten Holstein cows, naturally infected with S. aureus were used, with either early-stage or late-stage subclinical mastitis. Injection of rboGM-CSF caused a remarkable increase in milk CL activity with a peak at 6 h after the cytokine injection in the early- and late-stage groups. In the early-stage group, milk SCC stayed around preinjection level at 6 h, rose significantly on days 1 and 2, and was followed by a smooth and significant decline to an under preinjection level (below 200 000 cells/mL) on day 7 postinjection. Alternatively, in the late-stage group, milk SCC rose significantly at 6 h after the cytokine injection and maintained high levels thereafter. The milk S. aureus count decreased drastically by the cytokine injection in the early-stage group. The bacterial count was moderately decreased in the late-stage group, but increased back to preinoculation levels on day 7 after the cytokine injection. The results suggest that the rboGM-CSF has a potential as a therapeutic agent for S. aureus infection causing subclinical mastitis of dairy cows, if the cytokine is applied at the initial stage of infection. PMID:15352542

  10. Directional Fluid Transport across Organ-Blood Barriers: Physiology and Cell Biology.

    PubMed

    Caceres, Paulo S; Benedicto, Ignacio; Lehmann, Guillermo L; Rodriguez-Boulan, Enrique J

    2017-03-01

    Directional fluid flow is an essential process for embryo development as well as for organ and organism homeostasis. Here, we review the diverse structure of various organ-blood barriers, the driving forces, transporters, and polarity mechanisms that regulate fluid transport across them, focusing on kidney-, eye-, and brain-blood barriers. We end by discussing how cross talk between barrier epithelial and endothelial cells, perivascular cells, and basement membrane signaling contribute to generate and maintain organ-blood barriers.

  11. Highly potent stem cells from full-term amniotic fluid: A realistic perspective.

    PubMed

    Hamid, Adila A; Joharry, Muhammad Khair; Mun-Fun, Hoo; Hamzah, Siti Nurusaadah; Rejali, Zulida; Yazid, Mohd Nazri; Karuppiah, Thilakavathy; Nordin, Norshariza

    2017-03-01

    Amniotic fluid (AF) is now known to harbor highly potent stem cells, making it an excellent source for cell therapy. However, most of the stem cells isolated are from AF of mid-term pregnancies in which the collection procedure involves an invasive technique termed amniocentesis. This has limited the access in getting the fluid as the technique imposes certain level of risks to the mother as well as to the fetus. Alternatively, getting AF from full-term pregnancies or during deliveries would be a better resolution. Unfortunately, very few studies have isolated stem cells from AF at this stage of gestation, the fluid that is merely discarded. The question remains whether full-term AF harbors stem cells of similar potency as of the stem cells of mid-term AF. Here, we aim to review the prospect of having this type of stem cells by first looking at the origin and contents of AF particularly during different gestation period. We will then discuss the possibility that the AF, at full term, contains a population of highly potent stem cells. These stem cells are distinct from, and probably more potent than the AF mesenchymal stem cells (AF-MSCs) isolated from full-term AF. By comparing the studies on stem cells isolated from mid-term versus full-term AF from various species, we intend to address the prospect of having highly potent amniotic fluid stem cells from AF of full-term pregnancies in human and animals.

  12. Estimating the impact of somatic cell count on the value of milk utilising parameters obtained from the published literature.

    PubMed

    Geary, Una; Lopez-Villalobos, Nicolas; O'Brien, Bernadette; Garrick, Dorian J; Shalloo, Laurence

    2014-05-01

    The impact of mastitis on milk value per litre independent of the effect of mastitis on milk volume, was quantified for Ireland using a meta-analysis and a processing sector model. Changes in raw milk composition, cheese processing and composition associated with increased bulk milk somatic cell count (BMSCC) were incorporated into the model. Processing costs and market values were representative of current industry values. It was assumed that as BMSCC increased (i) milk fat and milk protein increased and milk lactose decreased, (ii) fat and protein recoveries decreased, (iii) cheese protein decreased and cheese moisture increased. Five BMSCC categories were examined from ⩽100 000 to >400 000 cells/ml. The analysis showed that as BMSCC increased the production quantities reduced. An increase in BMSCC from 100 000 to >400 000 cells/ml saw a reduction in net revenue of 3·2% per annum (€51·3 million) which corresponded to a reduction in the value of raw milk of €0·0096 cents/l.

  13. Propidium iodide (PI) stains Nissl bodies and may serve as a quick marker for total neuronal cell count.

    PubMed

    Niu, Junfei; Li, Chunman; Wu, Haihui; Feng, Xianling; Su, Qingning; Li, Shihe; Zhang, Lihong; Yew, David Tai Wai; Cho, Eric Yu Pang; Sha, Ou

    2015-03-01

    Propidium iodide (PI) reacts with both DNA and RNA and is a commonly used fluorescent reagent for nucleic acid staining. The aim of the study was to compare the cellular staining patterns of PI with that of Nissl staining in rat nervous tissues and to report a modified staining method that selectively labels Nissl bodies in neurons. Cryosections and paraffin sections of different tissues of normal Sprague-Dawley rats, including trigeminal ganglia, dorsal root ganglia, spinal cord, liver, and small intestine, were stained by either PI or the hematoxylin and eosin method. Some sections were treated with RNase or DNase before the above staining, and some were double stained with PI and a Nissl stain. The sections were observed by light, fluorescence or confocal microscopy. Results showed strong PI signals detected as patterns of granules in the neuronal cytoplasm of all nervous tissues, whereas the staining of neuronal nuclei was weaker. In contrast, nuclei of neuroglial cells were strongly stained by PI, while the cytoplasm was not obviously stained. Pretreatment of the neural tissue with RNase abolished the PI signals. Furthermore, the PI positive granules in neuronal cytoplasm co-localized with Nissl bodies stained by the fluorescent Nissl stain. When the tissue was pretreated with DNase, PI only stained the cytoplasmic granules of neurons, but not that of glial cells. Our results show that PI stains Nissl bodies and may serve as an economical and convenient neuron marker for neuronal cell counting when specific neural markers such as antibodies are not readily available.

  14. CD4+ T cell counts in initiation of antiretroviral therapy in HIV infected asymptomatic individuals; controversies and inconsistencies.

    PubMed

    Maina, E K; Bonney, E Y; Bukusi, E A; Sedegah, M; Lartey, M; Ampofo, W K

    2015-12-01

    The primary goal when devising strategies to define the start of therapy in HIV infected individuals is to avoid HIV disease progression and toxicity from antiretroviral therapy (ART). Intermediate goals includes, avoiding resistance by suppressing HIV replication, reducing transmission, limiting spread and diversity of HIV within the body and protecting the immune system from harm. The question of how early or late to start ART and achieve both primary and intermediate goals has dominated HIV research. The distinction between early and late treatment of HIV infection is currently a matter of CD4+ T cells count, a marker of immune status, rather than on viral load, a marker of virus replication. Discussions about respective benefits of early or delayed therapy, as well as the best CD4+ T cell threshold during the course of HIV infection at which ART is initiated remains inconclusive. Guidelines issued by various agencies, provide different initiation recommendations. This can be confusing for clinicians and policy-makers when determining the best time to initiate therapy. Optimizing ART initiation strategies are clearly complex and must be balanced between individual and broader public health needs. This review assesses available data that contributes to the debate on optimal time to initiate therapy in HIV-infected asymptomatic individuals. We also review reports on CD4+ T cell threshold to guide initiation of ART and finally discuss arguments for and against early or late initiation of ART.

  15. Non-Newtonian fluid effects on surface reactions in a microfluidic flow cell

    NASA Astrophysics Data System (ADS)

    Akgül, M. Bahattin; Sarı, Gözde; Pakdemirli, Mehmet

    2012-11-01

    Mass transfer over a reactive surface in microfluidic flow cells plays a key role in understanding biomoleculer interactions and diagnosis of small molecules for biomedical and environmental applications. The effects of Non-Newtonian power law fluid on the binding reaction kinetic of immunoglobulin G in a flow cell are analyzed in this study. Governing equations for the fluid flow, mass transport and surface reaction are derived. The finite element method is employed to solve resulting equations. In addition, the effects of volumetric flow rate, fluid behavior index and reaction constants on the surface reaction are analyzed and presented graphically.

  16. 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.

  17. The importance of endometrial nerve fibers and macrophage cell count in the diagnosis of endometriosis

    PubMed Central

    Cetin, Cihan; Serdaroglu, Hasan; Tuzlali, Sitki

    2013-01-01

    Background: Endometriosis is a disease that is hard to diagnose without the gold standard method, laparoscopy. An easier diagnostic method is needed. Objective: The aim of the study is to determine whether the number of macrophage cells in the endometrium and/or the detection of nerve fibers can be used in the diagnosis of endometriosis. Materials and Methods: Endometrial sampling was done to 31 patients prior to laparoscopy (L/S) or laparotomy (L/T) at Istanbul University Istanbul School of Medicine Hospital between January 2010 February 2011. Also 34 patients who were retrospectively chosen from their files were added to the study. 5 patients were excluded from the study. Totally, 31 patients were placed in the endometriosis and 29 patients in the control group. Endometrial samples were evaluated immunohistochemically with the markers protein gene product 9.5 (PGP 9.5) and neurofilament (NF) for nerve fibers and CD68 for macrophages. Results: None of the samples were stained with PGP 9.5 and NF. As for CD68+cells, no statistically significant difference was observed between groups (endometriosis: 216.10±104.41; control: 175.93±43.05, p=0.06). Results were also evaluated in the subgroups of menstruel phases and disease stages. Only in the proliferative phase there was a significant increase in the endometriosis group (p=0.03). No significant difference was observed between the stages. Conclusion: The detection of nerve fibers in the eutopic endometrium with the markers of PGP 9.5 and NF is not found to be helpful in the diagnosis of endometriosis. Macrophage cells may be helpful in the diagnosis only in the proliferative phase. PMID:24639773

  18. Determinants, reproducibility, and seasonal variation of bacterial cell wall components and viable counts in house dust.

    PubMed

    Leppänen, H K; Täubel, M; Roponen, M; Vepsäläinen, A; Rantakokko, P; Pekkanen, J; Nevalainen, A; von Mutius, E; Hyvärinen, A

    2015-06-01

    The objectives of this study were (i) to assess the determinants that affect concentrations of the bacterial cell wall components 3-hydroxy fatty acids (3-OH FAs) and muramic acid and of total viable bacteria and actinomycetes in house dust; and (ii) to examine the seasonal variation and reproducibility of these bacterial cell wall components in house dust. A number of lifestyle and environmental factors, mostly not consistent for different bacterial measures but commonly including the type of dwelling and farming (number of livestock), explained up to 37% of the variation of the bacterial concentrations in 212 homes in Eastern Finland. The reproducibility of 3-OH FAs and muramic acid measurements in house dust were studied in five urban homes and were found to be generally high (ICC 74-84%). Temporal variation observed in repeated sampling of the same home throughout a year was more pronounced for 3-OH FAs determinations (ICC 22%) than for muramic acid (ICC 55-66%). We conclude that determinants vary largely for different types of bacterial measurements in house dust; the measured parameters represent different aspects of the bacterial content indoors. More than one sample is needed to describe bacterial concentrations in house dust in the home environment due to large temporal variation.

  19. Suppression of Dendritic Cell Maturation and T Cell Proliferation by Synovial Fluid Myeloid Cells from Mice with Autoimmune Arthritis

    PubMed Central

    Egelston, Colt; Kurkó, Júlia; Besenyei, Timea; Tryniszewska, Beata; Rauch, Tibor A.; Glant, Tibor T.; Mikecz, Katalin

    2012-01-01

    Objective To determine whether myeloid cells (such as granulocytes) present in the synovial fluid (SF) of arthritic joints have an impact on adaptive immunity. Specifically, we investigated the effects of SF cells, harvested from the joints of mice with proteoglycan (PG)-induced arthritis (PGIA), on dendritic cell (DC) maturation and antigen-specific T-cell proliferation. Methods We monitored DC maturation (MHC class II and CD86 expression) by flow cytometry upon co-culture of DCs with SF or spleen myeloid cells from mice with PGIA. The effects of these myeloid cells on T-cell proliferation were studied using T cells purified from PG-specific T cell receptor transgenic (PG-TCR-Tg) mice. Phenotypic analysis of myeloid cells was performed employing immunostaining, RT-PCR, Western blot, and biochemical assays. Results Inflammatory SF cells significantly suppressed the maturation of DCs upon co-culture. PG-TCR-Tg T cells cultured with antigen-loaded DCs showed dramatic decreases in proliferation in the presence of SF cells. Spleen myeloid cells from arthritic mice did not have suppressive effects. SF cells were unable to suppress CD3/CD28-stimulated proliferation of the same T cells, suggesting a DC-dependent mechanism. SF cells exhibited all of the characteristics of myeloid-derived suppressor cells (MDSCs), and exerted suppression primarily through production of nitric oxide and reactive oxygen species by granulocyte-like cells. Conclusion SF in the joints of mice with PGIA contains a population of granulocytic MDSCs that potently suppress DC maturation and T-cell proliferation. These MDSCs have the potential to limit the expansion of autoreactive T cells, thus breaking the vicious cycle of autoimmunity and inflammation. PMID:22492217

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

    SciTech Connect

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

    1995-10-01

    The use of ion chromatography (IC) for radiochemical separations is a well established technique. IC is commonly used in routine environmental monitoring applications as well as in specialized research applications. Typical usage involves the separation of a single radionuclide from the non-radioactive constituents. During the past decade, a limited amount of research has been conducted using automated IC systems in actinide separation applications (e.g.). More recently, separation procedures for common non-gamma emitting activation and fission products were developed utilizing a high performance liquid chromatography (HPLC) system. In addition, a separation procedure for six common actinides has been developed using a HPLC system. These latter systems used on-line flow-cell detectors for quantification of the radioactive constituents of the effluent stream.

  1. High interstitial fluid pressure promotes tumor cell proliferation and invasion in oral squamous cell carcinoma.

    PubMed

    Yu, Tao; Liu, Kun; Wu, Yingying; Fan, Jinchuan; Chen, Jianchao; Li, Chao; Zhu, Guiquan; Wang, Zhaohui; Li, Longjiang

    2013-11-01

    It has been shown that interstitial fluid pressure (IFP) is elevated in many solid tumors. The elevated IFP in tumors is responsible, at least in part, for the poor blood supply, inadequate delivery of therapeutic agents to solid tumors and poor treatment response in patients. The present study was carried out to examine alterations in malignant phenotypes in oral squamous cell carcinoma cells subjected to conditions mimicking IFP and to identify the relevant molecular mechanisms. We investigated tumor cell proliferation and invasion using SCC-4 and SCC-9 cells subjected to an increased extracellular pressure of 0, 15 and 30 mmHg in vitro. The results revealed that the increased IFP resulted in a marked increase in cancer cell proliferation, survival and invasion in vitro and altered the expression of >1,800 genes involved in invasion and metastasis, the heat shock pathway, the p38 and JNK signaling pathway, apoptosis and the cell growth and differentiation signaling pathway. These results suggest the important potential clinical application of measuring IFP, which can be used as a generic marker of prognosis and response to therapy.

  2. Association between somatic cell count during the first lactation and the cumulative milk yield of cows in Irish dairy herds.

    PubMed

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

    2014-01-01

    Reduced potential milk yield is an important component of mastitis costs in dairy cows. The first aim of this study was to assess associations between somatic cell count (SCC) during the first lactation, and cumulative milk yield over the first lactation and subsequent lifetime of cows in Irish dairy herds. The second aim was to assess the association between SCC at 5 to 30d in milk during parity 1 (SCC1), and SCC over the entire first lactation for cows in Irish dairy herds. The data set studied included records from 51,483 cows in 5,900 herds. Somatic cell count throughout the first lactation was summarized using the geometric mean and variance of SCC. Data were analyzed using linear models that included random effects to account for the lack of independence between observations, and herd-level variation in coefficients. Models were developed in a Bayesian framework and parameters were estimated from 10,000 Markov chain Monte Carlo simulations. The final models were a good fit to the data. A 1-unit increase in mean natural logarithm SCC over the first lactation was associated with a median decrease in first lactation and lifetime milk yield of 135 and 1,663kg, respectively. A 1-unit increase in the variance of natural logarithm SCC over the first lactation was associated with a median decrease in lifetime milk yield of 719kg. To demonstrate the context of lifetime milk yield results, microsimulation was used to model the trajectory of individual cows and evaluate the expected outcomes for particular changes in herd-level geometric mean SCC over the first lactation. A 75% certainty of savings of at least €199/heifer in the herd was detected if herd-level geometric mean SCC over the first lactation was reduced from ≥120,000 to ≤72,000cells/mL. The association between SCC1 and SCC over the remainder of the first lactation was highly herd dependent, indicating that control measures for heifer mastitis should be preferentially targeted on an individual

  3. Online quantitative phase imaging of vascular endothelial cells under fluid shear stress utilizing digital holographic microscopy

    NASA Astrophysics Data System (ADS)

    Odenthal-Schnittler, Maria; Schnittler, Hans Joachim; Kemper, Björn

    2016-03-01

    We have explored the utilization of quantitative phase imaging with digital holographic microscopy (DHM) as a novel tool for quantifying the dynamics of morphologic parameters (morphodynamics) of confluent endothelial cell layers under fluid shear stress conditions. Human umbilical vein endothelial cells (HUVECs) were exposed to fluid shear stress in a transparent cone/plate flow device (BioTech-Flow-System) and imaged with a modular setup for quantitative DHM phase imaging for up to 48 h. The resulting series of quantitative phase image sequences were analyzed for the average surface roughness of the cell layers and cell alignment. Our results demonstrate that quantitative phase imaging is a powerful and reliable tool to quantify the dynamics of morphological adaptation of endothelial cells to fluid shear stress.

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

    PubMed

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

    2011-07-01

    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 produced more mineralized matrix but delayed peaks in osteogenic markers. Cells were also cultured on 3D scaffolds constructed of poly-ε-caprolactone for 15 weeks. MSCs differentiated more quickly than AFS cells on 3D scaffolds, but mineralized matrix production slowed considerably after 5 weeks. In contrast, the rate of AFS cell mineralization continued to increase out to 15 weeks, at which time AFS constructs contained 5-fold more mineralized matrix than MSC constructs. Therefore, cell source should be taken into consideration when used for cell therapy, as the MSCs would be a good choice for immediate matrix production, but the AFS cells would continue robust mineralization for an extended period of time. This study demonstrates that stem cell source can dramatically influence the magnitude and rate of osteogenic differentiation in vitro.

  5. 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.

  6. Absolute lymphocyte count as predictor of overall survival for patients with multiple myeloma treated with single autologous stem cell transplant.

    PubMed

    Jimenez-Zepeda, Victor H; Reece, Donna E; Trudel, Suzanne; Chen, Christine; Franke, Norman; Winter, Andrew; Tiedemann, Rodger; Kukreti, Vishal

    2015-01-01

    Post-autologous stem cell transplant (ASCT) studies have demonstrated that absolute lymphocyte count (ALC) recovery is associated with prolonged survival in some hematological malignancies. To assess whether ALC recovery has prognostic significance in patients with multiple myeloma (MM) undergoing single ASCT, we conducted a retrospective analysis of ALC at different time-points in patients with MM. In total 769 consecutive patients who underwent single ASCT from January 2000 to December 2007 were evaluated. An ALC of ≥ 1400 cells/μL at day 0, day 15 and day 90 significantly correlated with a better overall survival (OS) (median OS of 111, 90.7 and 84 months vs. 74, 70.5 and 65 months, respectively, p < 0.001 for all time-points). Multivariate analysis showed that ALC is an independent prognostic factor for OS after ASCT. In conclusion, ALC is a surrogate marker of the host immune system that correlates with better survival in patients with MM undergoing single ASCT. Immunomodulatory drugs, vaccination strategies and cellular therapies in MM should be investigated.

  7. Protective effect of curcumin on experimentally induced arthritic rats: detailed histopathological study of the joints and white blood cell count.

    PubMed

    Kamarudin, Taty Anna; Othman, Faizah; Mohd Ramli, Elvy Suhana; Md Isa, Nurismah; Das, Srijit

    2012-01-01

    Curcuma longa (turmeric) rhizomes contains curcumin, an active compound which possesses anti-inflammatory effects. Collagen-induced arthritis (CIA) is an accepted experimental animal model of rheumatoid arthritis. The present study aimed to observe the histological changes in the joints of experimental arthritic rats treated with curcumin. Twenty four male Sprague-Dawley (approximately 7 weeks-old) rats were randomly divided into four groups. Three groups were immunized with 150 µg collagen. All rats with established CIA, with arthritis scores exceeding 1, were orally treated with betamethasone (0.5 mg/ml/kg body weight), curcumin (110 mg/ml/kg body weight) or olive oil (1.0 ml/kg body weight) daily, for two weeks. One remaining group was kept as normal control. Treatment with 110 mg/ml/kg curcumin showed significant mean differences in the average white blood cell (WBC) count (p<0.05), cell infiltration, bone and cartilage erosion scores (p<0.05) compared to the olive oil treated group. Pannus formation scores showed that curcumin supplementation successfully suppressed the pannus formation process that occurred in the articular cartilage of the CIA joints. The mean difference for histological scores for the curcumin group was insignificant compared to the betamethasone treated group. It is concluded that supplementation of curcumin has protective effect on the histopathological and degenerative changes in the joints of CIA rats which was at par with betamethasone.

  8. Emergent long-range couplings in arrays of fluid cells

    SciTech Connect

    Abraham, Douglas Bruce

    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.

  9. 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.

  10. 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

  11. Effect of Nadir CD4+ T Cell Count on Clinical Measures of Periodontal Disease in HIV+ Adults before and during Immune Reconstitution on HAART

    PubMed Central

    Vernon, Lance T.; Demko, Catherine A.; Babineau, Denise C.; Wang, Xuelei; Toossi, Zahra; Weinberg, Aaron; Rodriguez, Benigno

    2013-01-01

    Background The contribution of HIV-infection to periodontal disease (PD) is poorly understood.  We proposed that immunological markers would be associated with improved clinical measures of PD. Methods We performed a longitudinal cohort study of HIV-infected adults who had started highly active antiretroviral therapy (HAART) <2 years. PD was characterized clinically as the percent of teeth with ≥1 site with periodontal probing depth (PPD) ≥5.0mm, recession (REC) >0mm, clinical attachment level (CAL) ≥4.0mm, and bleeding on probing (BOP) at ≥4 sites/tooth and microbiologically as specific periodontopathogen concentration. Linear mixed-effects models were used to assess the associations between immune function and PD. Results Forty (40) subjects with median 2.7 months on HAART and median nadir CD4+ T-cell count of 212 cells/μl completed a median 3 visits. Over 24 months, CD4+ T-cell count increased by a mean 173 cells/µl (p<0.001) and HIV RNA decreased by 0.5 log10 copies/ml (p<0.001); concurrently, PPD, CAL and BOP decreased by a mean 11.7%, 12.1%, and 14.7% respectively (all p<0.001). Lower nadir CD4+ T-cell count was associated with worse baseline REC (-6.72%; p=0.04) and CAL (9.06%; p<0.001). Further, lower nadir CD4+ T-cell count was associated with a greater relative longitudinal improvement in PPD in subjects with higher baseline levels of Porphyromonas gingivalis (p=0.027), and BOP in subjects with higher baseline levels of Porphyromonas gingivalis or Treponema denticola (p=0.001 and p=0.006 respectively). Longitudinal changes from baseline in CD4+ T-cell count and level of HIV RNA were not independently associated with longitudinal changes in any clinical markers of PD. Conclusion Degree of immunosuppression was associated with baseline gingival recession. After HAART initiation, measures of active PD improved most in those with lower nadir CD4+ T-cell counts and higher baseline levels of specific periodontopathogens. Nadir CD4+ T-cell count

  12. Cell distribution in a scaffold with random architectures under the influence of fluid dynamics.

    PubMed

    Shanglong Xu; Pingan Du; Youzhuan Xie; Yang Yue

    2008-11-01

    Fluid dynamic environment and scaffold architectures have an important influence on cell growth and distribution inside the scaffold. A porous cylindrical scaffold with a central channel is seeded with the sheep mesenchymal stem cells (MSCs) in this study. Then the cell seeded scaffold is continuously perfused with alpha-MEM medium by a peristaltic pump for 7, 14, and 28 days. Histological study shows that the cell proliferation rates are different throughout the whole scaffolds. The different cell coverage is shown in various positions of the scaffold. A computational fluid dynamics (CFD) modeling is used to simulate the flow conditions within perfused cell-seeded scaffolds to give insight into the mechanisms of these cell growth phenomena. Relating the simulation results to perfusion experiments, the even fluid velocity (approximately 0.26-0.64 mm/s) and shear stress (approximately 0.0029-0.027 Pa) are found to correspond to increased cell proliferation within the cell-scaffold constructs. This method exhibits novel capabilities to compare results obtained for different perfusion rates or different scaffold microarchitectures and may allow specific fluid velocities and shear stresses to be determined that optimize the perfusion flow rate, porous scaffold architecture, and distribution of in vitro tissue growth.

  13. Fluid flow facilitates inward rectifier K+ current by convectively restoring [K+] at the cell membrane surface

    PubMed Central

    Kim, Jae Gon; Park, Sang Woong; Byun, Doyoung; Choi, Wahn Soo; Sung, Dong Jun; Shin, Kyung Chul; Kim, Hyun-ji; Leem, Young-Eun; Kang, Jong-Sun; Cho, Hana; Kim, Bokyung; Cho, Sung I; Bae, Young Min

    2016-01-01

    The inward rectifier Kir2.1 current (IKir2.1) was reported to be facilitated by fluid flow. However, the mechanism underlying this facilitation remains uncertain. We hypothesized that during K+ influx or efflux, [K+] adjacent to the outer mouth of the Kir2.1 channel might decrease or increase, respectively, compared with the average [K+] of the bulk extracellular solution, and that fluid flow could restore the original [K+] and result in the apparent facilitation of IKir2.1. We recorded the IKir2.1 in RBL-2H3 cells and HEK293T cells that were ectopically over-expressed with Kir2.1 channels by using the whole-cell patch-clamp technique. Fluid-flow application immediately increased the IKir2.1, which was not prevented by either the pretreatment with inhibitors of various protein kinases or the modulation of the cytoskeleton and caveolae. The magnitudes of the increases of IKir2.1 by fluid flow were driving force-dependent. Simulations performed using the Nernst-Planck mass equation indicated that [K+] near the membrane surface fell markedly below the average [K+] of the bulk extracellular solution during K+ influx, and, notably, that fluid flow restored the decreased [K+] at the cell surface in a flow rate-dependent manner. These results support the “convection-regulation hypothesis” and define a novel interpretation of fluid flow-induced modulation of ion channels. PMID:28004830

  14. In Vitro Bone Cell Models: Impact of Fluid Shear Stress on Bone Formation

    PubMed Central

    Wittkowske, Claudia; Reilly, Gwendolen C.; Lacroix, Damien; Perrault, Cecile M.

    2016-01-01

    This review describes the role of bone cells and their surrounding matrix in maintaining bone strength through the process of bone remodeling. Subsequently, this work focusses on how bone formation is guided by mechanical forces and fluid shear stress in particular. It has been demonstrated that mechanical stimulation is an important regulator of bone metabolism. Shear stress generated by interstitial fluid flow in the lacunar-canalicular network influences maintenance and healing of bone tissue. Fluid flow is primarily caused by compressive loading of bone as a result of physical activity. Changes in loading, e.g., due to extended periods of bed rest or microgravity in space are associated with altered bone remodeling and formation in vivo. In vitro, it has been reported that bone cells respond to fluid shear stress by releasing osteogenic signaling factors, such as nitric oxide, and prostaglandins. This work focusses on the application of in vitro models to study the effects of fluid flow on bone cell signaling, collagen deposition, and matrix mineralization. Particular attention is given to in vitro set-ups, which allow long-term cell culture and the application of low fluid shear stress. In addition, this review explores what mechanisms influence the orientation of collagen fibers, which determine the anisotropic properties of bone. A better understanding of these mechanisms could facilitate the design of improved tissue-engineered bone implants or more effective bone disease models. PMID:27896266

  15. Fluid flow facilitates inward rectifier K(+) current by convectively restoring [K(+)] at the cell membrane surface.

    PubMed

    Kim, Jae Gon; Park, Sang Woong; Byun, Doyoung; Choi, Wahn Soo; Sung, Dong Jun; Shin, Kyung Chul; Kim, Hyun-Ji; Leem, Young-Eun; Kang, Jong-Sun; Cho, Hana; Kim, Bokyung; Cho, Sung I; Bae, Young Min

    2016-12-22

    The inward rectifier Kir2.1 current (IKir2.1) was reported to be facilitated by fluid flow. However, the mechanism underlying this facilitation remains uncertain. We hypothesized that during K(+) influx or efflux, [K(+)] adjacent to the outer mouth of the Kir2.1 channel might decrease or increase, respectively, compared with the average [K(+)] of the bulk extracellular solution, and that fluid flow could restore the original [K(+)] and result in the apparent facilitation of IKir2.1. We recorded the IKir2.1 in RBL-2H3 cells and HEK293T cells that were ectopically over-expressed with Kir2.1 channels by using the whole-cell patch-clamp technique. Fluid-flow application immediately increased the IKir2.1, which was not prevented by either the pretreatment with inhibitors of various protein kinases or the modulation of the cytoskeleton and caveolae. The magnitudes of the increases of IKir2.1 by fluid flow were driving force-dependent. Simulations performed using the Nernst-Planck mass equation indicated that [K(+)] near the membrane surface fell markedly below the average [K(+)] of the bulk extracellular solution during K(+) influx, and, notably, that fluid flow restored the decreased [K(+)] at the cell surface in a flow rate-dependent manner. These results support the "convection-regulation hypothesis" and define a novel interpretation of fluid flow-induced modulation of ion channels.

  16. Absence of trisomy 7 in nonneoplastic human ascitic and pleural fluid cells. An interphase cytogenetic study.

    PubMed

    Larramendy, M L; Björkqvist, A M; Tammilehto, L; Taavitsainen, M; Mattson, K; Knuutila, S

    1994-11-01

    Trisomy 7 is a frequent aneuploid change in lymphomas, adenocarcinomas, and malignant mesenchymal and neurogenic tumors. Moreover, it has been observed in cultured and uncultured non-neoplastic cells from brain, kidney, liver, lung, and atherosclerotic plaques, among other tissues, opening debate on the role of this change in normal and neoplastic tissue. We used nonradioactive in situ hybridization (ISH) with a biotinylated chromosome 7-specific alpha-satellite DNA probe to seek an extra copy of chromosome 7 in ascitic and pleural fluid interphase cells from 26 donors. The donors comprised 24 patients with nonmalignant clinical history, one patient with non-Hodgkin's malignant lymphoma (positive control), and one patient with chronic myeloid leukemia (CML, negative control). The highest frequency of fluid cells with three hybridization signals in patients without neoplasia was 0.5%, in contrast to the frequency of 40.5% noted in the fluid cells of the patient with non-Hodgkin's malignant lymphoma. The results demonstrate that the frequency of trisomic cells in pleural as well as in ascitic fluid is very low, making possible use of the cells in ascitic or pleural fluids in identification of malignancy.

  17. Cooperative effects of matrix stiffness and fluid shear stress on endothelial cell behavior.

    PubMed

    Kohn, Julie C; Zhou, Dennis W; Bordeleau, François; Zhou, Allen L; Mason, Brooke N; Mitchell, Michael J; King, Michael R; Reinhart-King, Cynthia A

    2015-02-03

    Arterial hemodynamic shear stress and blood vessel stiffening both significantly influence the arterial endothelial cell (EC) phenotype and atherosclerosis progression, and both have been shown to signal through cell-matrix adhesions. However, the cooperative effects of fluid shear stress and matrix stiffness on ECs remain unknown. To investigate these cooperative effects, we cultured bovine aortic ECs on hydrogels matching the elasticity of the intima of compliant, young, or stiff, aging arteries. The cells were then exposed to laminar fluid shear stress of 12 dyn/cm(2). Cells grown on more compliant matrices displayed increased elongation and tighter EC-cell junctions. Notably, cells cultured on more compliant substrates also showed decreased RhoA activation under laminar shear stress. Additionally, endothelial nitric oxide synthase and extracellular signal-regulated kinase phosphorylation in response to fluid shear stress occurred more rapidly in ECs cultured on more compliant substrates, and nitric oxide production was enhanced. Together, our results demonstrate that a signaling cross talk between stiffness and fluid shear stress exists within the vascular microenvironment, and, importantly, matrices mimicking young and healthy blood vessels can promote and augment the atheroprotective signals induced by fluid shear stress. These data suggest that targeting intimal stiffening and/or the EC response to intima stiffening clinically may improve vascular health.

  18. Elevated granzyme B+ B-cell level in SIV-infection correlate with viral load and low CD4 T-cell count

    PubMed Central

    Kotb, Ahmad; Klippert, Antonina; Daskalaki, Maria; Sauermann, Ulrike; Stahl-Hennig, Christiane; Neumann, Berit

    2017-01-01

    Granzyme B-expressing (GrB+) B cells are thought to contribute to immune dysfunctions in HIV patients, but so far their exact role is unknown. This report demonstrates for the first time the existence of GrB+ B cells in SIV-infected rhesus macaques, which represent the most commonly used nonhuman primate model for HIV research. Similar to HIV patients, we found significantly higher frequencies of these cells in the blood of chronically SIV-infected rhesus monkeys compared with uninfected healthy ones. These frequencies correlated with plasma viral load and inversely with absolute CD4 T-cell counts. When investigating GrB+ B cells in different compartments, levels were highest in blood, spleen and bone marrow, but considerably lower in lymph nodes and tonsils. Analysis of expression of various surface markers on this particular B-cell subset in SIV-infected macaques revealed differences between the phenotype in macaques and in humans. GrB+ B cells in SIV-infected rhesus macaques exhibit an elevated expression of CD5, CD10, CD25 and CD27, while expression of CD19, CD185 and HLA-DR is reduced. In contrast to human GrB+ B cells, we did not observe a significantly increased expression of CD43 and CD86. B-cell receptor stimulation in combination with IL-21 of purified B cells from healthy animals led to the induction of GrB expression. Furthermore, initial functional analyses indicated a regulatory role on T-cell proliferation. Overall, our data pave the way for longitudinal analyses including studies on the functionality of GrB+ B cells in the nonhuman primate model for AIDS. PMID:27779180

  19. Retail display evaluation of steaks from select beef strip loins injected with a brine containing 1% ammonium hydroxide. Part 1: Fluid loss, oxidation, color, and microbial plate counts.

    PubMed

    Parsons, A N; VanOverbeke, D L; Goad, C L; Mireles Dewitt, C A

    2011-01-01

    Select beef loin pairs (n = 10) were injected (10% pump) with brine containing either 4.5% sodium-based phosphates, (CON), or 1% ammonium hydroxide treatment (AHT). Both brines also contained 3.6% NaCl and 1% Rosemary Herbalox. Steaks cut from loins were high oxygen (80% O(2)/20% CO(2)) modified atmosphere packaged, stored 4 d at 4 °C in the dark to simulate transportation, and then placed in retail display for 14 d (4 °C). On day 0, 7, and 14 of retail display steak properties were measured. Purge from AHT steaks was higher than CON (P < 0.05). Panelists were not able to visually discriminate between AHT and CON steaks through the first 6 d of retail display. After day 6, panelists rated AHT steaks higher for muscle color, percent discoloration, and overall color. Steaks from both treatments started at day 0 retail display with similar total plate counts (P > 0.05). Microbial counts increased more rapidly for AHT steaks than CON steaks (P < 0.05). AHT and CON steaks were not different in terms of lipid oxidation through day 7 retail display. By day 14 retail display CON steaks were above the threshold for consumer perception of oxidized flavors in fresh meat. However, results also indicated the AHT and CON steaks were no longer acceptable by day 14 in terms of color, were questionable in terms of microbial load, and likely were beyond their reasonable shelf life. Based on retail display properties, results indicated 1% AHT could successfully replace 4.5% SP in a meat injection brine. Practical Application: The research in this report compares steaks that have been injected with a commercial brine formulated with SP to steaks that have been injected with a brine, where the SP in the formulation are replaced with 1% AHT. Ammonium hydroxide is an USDA-FSIS approved ingredient in brines injected into fresh meats. Successful replacement of sodium phosphate with ammonium hydroxide would allow processors to significantly reduce the sodium content of injected fresh meat.

  20. An update clinical application of amniotic fluid-derived stem cells (AFSCs) in cancer cell therapy and tissue engineering.

    PubMed

    Gholizadeh-Ghaleh Aziz, Shiva; Fathi, Ezzatollah; Rahmati-Yamchi, Mohammad; Akbarzadeh, Abolfazl; Fardyazar, Zahra; Pashaiasl, Maryam

    2017-06-01

    Recent studies have elucidated that cell-based therapies are promising for cancer treatments. The human amniotic fluid stem (AFS) cells are advantageous cells for such therapeutic schemes that can be innately changed to express therapeutic proteins. HAFSCs display a natural tropism to cancer cells in vivo. They can be useful in cancer cells targeting. Moreover, they are easily available from surplus diagnostic samples during pregnancy and less ethical and legal concern are associated with the collection and application than other putative cells are subjected. This review will designate representatives of amniotic fluid and stem cell derived from amniotic fluid. For this propose, we collect state of human AFS cells data applicable in cancer therapy by dividing this approach into two main classes (nonengineered and engineered based approaches). Our study shows the advantage of AFS cells over other putative cells types in terms differentiation ability to a wide range of cells by potential and effective use in preclinical studies for a variety of diseases. This study has shown the elasticity of human AFS cells and their favorable potential as a multipotent cell source for regenerative stem cell therapy and capable of giving rise to multiple lineages including such as osteoblasts and adipocyte.

  1. 46,XY/45,X mosaicism in an amniotic fluid cell culture: suppression of abnormal cell line after subcultivation.

    PubMed Central

    Hasholt, L

    1979-01-01

    An abnormal cell population, 45,X, appeared in 3 of 4 cell lines established from an amniotic fluid specimen obtained from a normal mid-trimester pregnancy. Two of the cell lines were subjected to repeated chromosome analyses until VII passage. The abnormal cells were suppressed after repeated trypsinisations; simultaneously, fibroblast-like cells outgrew the cultures, which were previously predominated by epithelial-like cells. Polyploidy was found in 0 to 12% of the cells, the highest level existing in the early passages. The question of whether chromosomally abnormal cells present in primary cultures and the early subcultures reflect the karyotype of the fetus is discussed. PMID:573801

  2. Somatic Cells Count and Its Genetic Association with Milk Yield in Dairy Cattle Raised under Thai Tropical Environmental Conditions.

    PubMed

    Jattawa, D; Koonawootrittriron, S; Elzo, M A; Suwanasopee, T

    2012-09-01

    Somatic cells count (SCC), milk yield (MY) and pedigree information of 2,791 first lactation cows that calved between 1990 and 2010 on 259 Thai farms were used to estimate genetic parameters and trends for SCC and its genetic association with MY. The SCC were log-transformed (lnSCC) to make them normally distributed. An average information-restricted maximum likelihood procedure was used to estimate variance components. A bivariate animal model that considered herd-yr-season, calving age, and regression additive genetic group as fixed effects, and animal and residual as random effects was used for genetic evaluation. Heritability estimates were 0.12 (SE = 0.19) for lnSCC, and 0.31 (SE = 0.06) for MY. The genetic correlation estimate between lnSCC and MY was 0.26 (SE = 0.59). Mean yearly estimated breeding values during the last 20 years increased for SCC (49.02 cells/ml/yr, SE = 26.81 cells/ml/yr; p = 0.08), but not for MY (0.37 kg/yr, SE = 0.87 kg/yr; p = 0.68). Sire average breeding values for SCC and MY were higher than those of cows and dams (p<0.01). Heritability estimates for lnSCC and MY and their low but positive genetic correlation suggested that selection for low SCC may be feasible in this population as it is in other populations of dairy cows. Thus, selection for high MY and low SCC should be encouraged in Thai dairy improvement programs to increase profitability by improving both cow health and milk yield.

  3. 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.

  4. Pyogenic arthritis, pyoderma gangrenosum, and acne (PAPA) syndrome: differential diagnosis of septic arthritis by regular detection of exceedingly high synovial cell counts.

    PubMed

    Löffler, W; Lohse, P; Weihmayr, T; Widenmayer, W

    2017-03-01

    Pyogenic arthritis, pyoderma gangrenosum and acne syndrome was diagnosed in a 42-year-old patient, after an unusual persistency of high synovial cell counts had been noticed. Clinical peculiarities and problems with diagnosing septic versus non-septic arthritis are discussed.

  5. Mean cell volume of neutrophils and monocytes compared with C-reactive protein, interleukin-6 and white blood cell count for prediction of sepsis and nonsystemic bacterial infections.

    PubMed

    Mardi, D; Fwity, B; Lobmann, R; Ambrosch, A

    2010-08-01

    Clinicians are in need of better diagnostic markers for rapid diagnosis of severe infections. Therefore, we studied the diagnostic significance of mean cell volume of neutrophils (MNV) and monocytes (MMV) compared with Interleukin-6 (IL-6), C-reactive protein (CRP) and white blood cell count for predicting systemic clinical infection (sepsis). MNV and MMV were obtained by volume conductivity scatter (VCS) technique of the Coulter LH 750 hematology analyzer during automated differential counts, and blood samples from patients with sepsis (n = 37), nonsystemic bacterial infections (n = 39) and controls (n = 48) were studied. We observed a significant increase in MNV and MMV in the sepsis group compared with limited infections and controls. However, at a designated cut-off point of 250 pg/ml, IL-6 seemed to be the best predictor for sepsis with a sensitivity of 93% and a specificity of 76%. Compared with CRP (cut-off point 60 mg/dl), MNV at a cut-off of 150 had a comparable sensitivity and specificity and was the most predictive VCS parameter. Taken together, MNV and MMV seemed to be potential parameters to discriminate between sepsis and nonsystemic infections.

  6. First-line cART regimen impacts the course of CD8+ T-cell counts in HIV-infected patients that achieve sustained undetectable viral load.

    PubMed Central

    Poizot-Martin, Isabelle; Allavena, Clotilde; Delpierre, Cyrille; Duvivier, Claudine; Obry-Roguet, Véronique; Cano, Carla E.; Guillouet de Salvador, Francine; Rey, David; Dellamonica, Pierre; Cheret, Antoine; Cuzin, Lise; Katlama, Christine; Cabié, André; Hoen, Bruno

    2016-01-01

    Abstract The aim of the study was to investigate the impact of first-line combined antiretroviral therapy (cART) regimen on the course of CD8+ T-cell counts in human immunodeficiency virus (HIV)-infected patients. A retrospective observational study conducted on the French DAT’AIDS Cohort of HIV-infected patients. We selected 605 patients initiating a first-line cART between 2002 and 2009, and which achieved a sustained undetectable HIV plasma viral load (pVL) for at least 12 months without cART modification. The evolution of CD8+ T-cell counts according to cART regimen was assessed. CD8+ T-cell counts were assessed in 572 patients treated with 2NRTIs+1PI/r (n= 297), 2NRTIs+1NNRTI (n= 207) and 3NRTIs (n= 68). In multivariate analysis, after 12 months of follow-up, the 3NRTIs regimen was associated with a significantly smaller decrease of CD8+ T-cell count compared with NNRTI-containing regimens (–10.2 cells/μL in 3NRTIs vs –105.1 cells/μL; P=0.02) but not compared with PI-containing regimens (10.2 vs –60.9 cells/μL; P=0.21). After 24 months, the 3NRTIs regimen was associated with a smaller decrease of CD8+ T-cell count and % compared with PI/r- and NNRTI-containing regimens (0.2 in 3NRTIs vs –9.9 with PI/r-regimens, P=0.001, and vs –11.1 with NNRTI-regimens, p < 0.0001). A focus analysis on 11 patients treated with an INSTI-containing cART regimen during the study period showed after 12 months of follow-up, a median decrease of CD8+ T-cell count of –155 [inter quartile range: –302; –22] cells/μL. Our data highlight the fact that cART regimens have differential effects on CD8 pool down regulation. PMID:27741125

  7. A scanning acoustic microscope discriminates cancer cells in fluid

    NASA Astrophysics Data System (ADS)

    Miura, Katsutoshi; Yamamoto, Seiji

    2015-10-01

    Scanning acoustic microscopy (SAM) discriminates lesions in sections by assessing the speed of sound (SOS) or attenuation of sound (AOS) through tissues within a few minutes without staining; however, its clinical use in cytological diagnosis is unknown. We applied a thin layer preparation method to observe benign and malignant effusions using SAM. Although SAM is inferior in detecting nuclear features than light microscopy, it can differentiate malignant from benign cells using the higher SOS and AOS values and large irregular cell clusters that are typical features of carcinomas. Moreover, each single malignant cell exhibits characteristic cytoplasmic features such as a large size, irregular borders and secretory or cytoskeletal content. By adjusting the observation range, malignant cells are differentiated from benign cells easily using SAM. Subtle changes in the functional and structural heterogeneity of tumour cells were pursuable with a different digital data of SAM. SAM can be a useful tool for screening malignant cells in effusions before light microscopic observation. Higher AOS values in malignant cells compared with those of benign cells support the feasibility of a novel sonodynamic therapy for malignant effusions.

  8. Effect on quarter milk somatic cell count and antimicrobial susceptibility of Staphylococcus rostri causing intramammary infection in dairy water buffaloes.

    PubMed

    Locatelli, C; Piepers, S; De Vliegher, S; Barberio, A; Supré, K; Scaccabarozzi, L; Pisoni, G; Bronzo, V; Haesebrouck, F; Moroni, P

    2013-06-01

    In many parts of the world, coagulase-negative staphylococci (CNS) are the predominant cause of intramammary infections (IMI) in dairy cows and in water buffaloes, as well. A longitudinal field study was carried out on one well-managed dairy water buffalo herd to determine the prevalence and distribution of CNS and a recently described CNS-species, Staphylococcus rostri, in milk samples to explore its relevance for buffaloes' udder health throughout lactation, and to gain insight into the susceptibility of the latter species toward commonly used antimicrobials. Twice weekly quarter milk samples from a cohort of 11 lactating water buffaloes were collected over an 8-mo period. The CNS (n=109; 76.2% of all culture-positive samples) were the predominant pathogens causing IMI, followed by Corynebacterium bovis (n=11; 7.6%) and Streptococcus spp. (n=9; 6.2%) other than Stretococcus uberis (n=2; 1.4%). Thirty-seven hemolytic staphylococci suspected to be Staphylococcus aureus were further differentiated using transfer DNA-intergenic spacer-PCR and rpoB-gene sequencing because they were coagulase-negative. Thirty-three of those isolates were identified as Staph. rostri, whereas 2 others were identified as Staphylococcus epidermidis. None of the Staph. rostri isolates displayed resistance to the antimicrobial agents tested. Mean quarter milk somatic cell count (qSCC) of all samples collected throughout lactation was 20,970 cells/mL. The qSCC at sampling of quarters infected with Staph. rostri (34,466 cells/mL) and CNS other than Staph. rostri (34,813 cells/mL) were significantly higher than the qSCC of noninfected quarters (20,287 cells/mL), yet not significantly different from each other. These findings provide novel insight into the prevalence and distribution, antimicrobial susceptibility, and relevance of Staph. rostri compared with other CNS species causing IMI in water buffaloes. Further studies are needed to pinpoint the relevance, niches, and transmission routes of

  9. Ca²⁺ signaling and regulation of fluid secretion in salivary gland acinar cells.

    PubMed

    Ambudkar, Indu S

    2014-06-01

    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 Ca(2+) signaling processes as well as ion flux systems. Studies over the past four decades have demonstrated that Ca(2+) 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 [Ca(2+)] ([Ca(2+)]i) triggered by IP3-induced release of Ca(2+) 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 [Ca(2+)]i signal in the cell. However, Ca(2+) entry into the cell is required to sustain the elevation of [Ca(2+)]i and fluid secretion. This Ca(2+) 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, Ca(2+) entry via TRPC1 is a major determinant of fluid secretion. The receptor-evoked Ca(2+) 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 Ca(2+) signal can be ascribed to the polarized arrangement of the Ca(2+) channels, transporters, and signaling proteins. Distinct localization of these proteins in the cell suggests compartmentalization of Ca(2+) signals during regulation of fluid secretion. This chapter will discuss new concepts and findings regarding the polarization and control of Ca(2+) signals in the regulation of fluid secretion.

  10. Isolation of granulosa cells from follicular fluid; applications in biomedical and molecular biology experiments

    PubMed Central

    Aghadavod, Esmat; Zarghami, Nosratollah; Farzadi, Laya; Zare, Mina; Barzegari, Abolfazl; Movassaghpour, Ali Akbar; Nouri, Mohammad

    2015-01-01

    Background: Recently, a lot of research has been conducted to investigate the molecular mechanisms of the low quality of oocytes with granulosa cells (GCs). GCs are one of the major cell types found in follicular fluid and purification of these cells from the follicular fluid is very important for further studies. Although, there are different techniques of purification, a method for separation of highly-pure and minimally-damaged cells is necessary. In this paper, we presented a novel method for high purification of GCs with a large quantity and high purity. Materials and Methods: Follicular fluid was collected from patients who referred for in vitro fertilization and GCs in follicular fluid were extracted by Ficoll, Percoll and Red blood cell lysing buffer (RLB) methods. Then purity of extracted GCs was assessed by flow cytometry and morphological properties of GCs were observed by differential interference contrast microscopy. The purity of deoxyribonucleic acid and ribonucleic acid extracts was examined by NanoDrop 1000, pre-restriction fragment length polymorphism and electrophoresis techniques. Quality and quantity of extracting GCs were affected during the cell separation procedures. Results: Our results showed that each of purification method can affect quality and quantity of extracted cells. Conclusion: RLB method for extraction of GCs was shown to be a convenient procedure in comparison with Ficoll and Percoll methods. PMID:26918232

  11. Interstitial Fluid Flow: The Mechanical Environment of Cells and Foundation of Meridians

    PubMed Central

    Yao, Wei; Ding, Guanghong

    2012-01-01

    Using information from the deep dissection, microobservation, and measurement of acupoints in the upper and lower limbs of the human body, we developed a three-dimensional porous medium model to simulate the flow field using FLUENT software and to study the shear stress on the surface of interstitial cells (mast cells) caused by interstitial fluid flow. The numerical simulation results show the following: (i) the parallel nature of capillaries will lead to directional interstitial fluid flow, which may explain the long interstitial tissue channels or meridians observed in some experiments; (ii) when the distribution of capillaries is staggered, increases in the velocity alternate, and the velocity tends to be uniform, which is beneficial for substance exchange; (iii) interstitial fluid flow induces a shear stress, with magnitude of several Pa, on interstitial cell membranes, which will activate cells and lead to a biological response; (iv) capillary and interstitial parameters, such as capillary density, blood pressure, capillary permeability, interstitial pressure, and interstitial porosity, affect the shear stress on cell surfaces. The numerical simulation results suggest that in vivo interstitial fluid flow constitutes the mechanical environment of cells and plays a key role in guiding cell activities, which may explain the meridian phenomena and the acupuncture effects observed in experiments. PMID:23365601

  12. Study of Microfluidic System for Mechanical Property Measurement of Fluid-cell Interface

    NASA Astrophysics Data System (ADS)

    Moon, Ji Young; Lee, Jung Shin; Choi, Se Bin; Yoon, Hong Min; Tanner, Roger I.; Lee, Joon Sang

    2016-11-01

    The system for measuring the mechanical properties of active cell is studied through an integrated microfluidic system for cell separation, alignment and measurement of mechanical properties. A highly efficient lattice Boltzmann method (LBM) was employed to optimize the micro-fluidic system to investigate the interrelations between mechanical properties and various surrounding fluid ingredients which are difficult to observe using current experimental techniques. A combination model of the three dimensional LBM and the immersed boundary method (IBM) were used to simulate these systems. The LBM was used to determine incompressible fluid flow with a regular Eulerian grid. The IBM was used to solve the deformation of cells and matrix fluid interaction with a Lagrangian grid. Highly non-linear results such as cell-cell interactions, fluid-cell interactions, and optical force-cell interactions is studied. National Research Foundation of Korea (NRF) (Grant Number: NRF-2015R1A2A1A15056182, NRF-2015R1A5A1037668).

  13. T-cell count

    MedlinePlus

    ... 43. Read More Acute Acute lymphoblastic leukemia (ALL) HIV/AIDS Hodgkin lymphoma Immune response Immunodeficiency disorders Mononucleosis Multiple myeloma Radiation therapy Waldenstrom macroglobulinemia Review Date 8/29/2015 Updated by: Laura J. Martin, MD, MPH, ABIM ...

  14. CSF cell count

    MedlinePlus

    ... Fenichel GM, Jankovic J, Mazziotta JC, eds. Bradley's Neurology in Clinical Practice . 6th ed. Philadelphia, PA: Elsevier ... 1/2015 Updated by: Daniel Kantor, MD, Kantor Neurology, Coconut Creek, FL and immediate past president of ...

  15. Intradialytic aerobic cycling exercise alleviates inflammation and improves endothelial progenitor cell count and bone density in hemodialysis patients

    PubMed Central

    Liao, Min-Tser; Liu, Wen-Chih; Lin, Fu-Huang; Huang, Ching-Feng; Chen, Shao-Yuan; Liu, Chuan-Chieh; Lin, Shih-Hua; Lu, Kuo-Cheng; Wu, Chia-Chao

    2016-01-01

    Abstract Inflammation, endothelial dysfunction, and mineral bone disease are critical factors contributing to morbidity and mortality in hemodialysis (HD) patients. Physical exercise alleviates inflammation and increases bone density. Here, we investigated the effects of intradialytic aerobic cycling exercise on HD patients. Forty end-stage renal disease patients undergoing HD were randomly assigned to either an exercise or control group. The patients in the exercise group performed a cycling program consisting of a 5-minute warm-up, 20 minutes of cycling at the desired workload, and a 5-minute cool down during 3 HD sessions per week for 3 months. Biochemical markers, inflammatory cytokines, nutritional status, the serum endothelial progenitor cell (EPC) count, bone mineral density, and functional capacity were analyzed. After 3 months of exercise, the patients in the exercise group showed significant improvements in serum albumin levels, the body mass index, inflammatory cytokine levels, and the number of cells positive for CD133, CD34, and kinase insert domain-conjugating receptor. Compared with the exercise group, the patients in the control group showed a loss of bone density at the femoral neck and no increases in EPCs. The patients in the exercise group also had a significantly greater 6-minute walk distance after completing the exercise program. Furthermore, the number of EPCs significantly correlated with the 6-minute walk distance both before and after the 3-month program. Intradialytic aerobic cycling exercise programs can effectively alleviate inflammation and improve nutrition, bone mineral density, and exercise tolerance in HD patients. PMID:27399127

  16. 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

  17. 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

  18. Mounting Pressure in the Microenvironment: Fluids, Solids, and Cells in Pancreatic Ductal Adenocarcinoma.

    PubMed

    DuFort, Christopher C; DelGiorno, Kathleen E; Hingorani, Sunil R

    2016-06-01

    The microenvironment influences the pathogenesis of solid tumors and plays an outsized role in some. Our understanding of the stromal response to cancers, particularly pancreatic ductal adenocarcinoma, has evolved from that of host defense to tumor offense. We know that most, although not all, of the factors and processes in the microenvironment support tumor epithelial cells. This reappraisal of the roles of stromal elements has also revealed potential vulnerabilities and therapeutic opportunities to exploit. The high concentration in the stroma of the glycosaminoglycan hyaluronan, together with the large gel-fluid phase and pressures it generates, were recently identified as primary sources of treatment resistance in pancreas cancer. Whereas the relatively minor role of free interstitial fluid in the fluid mechanics and perfusion of tumors has been long appreciated, the less mobile, gel-fluid phase has been largely ignored for historical and technical reasons. The inability of classic methods of fluid pressure measurement to capture the gel-fluid phase, together with a dependence on xenograft and allograft systems that inaccurately model tumor vascular biology, has led to an undue emphasis on the role of free fluid in impeding perfusion and drug delivery and an almost complete oversight of the predominant role of the gel-fluid phase. We propose that a hyaluronan-rich, relatively immobile gel-fluid phase induces vascular collapse and hypoperfusion as a primary mechanism of treatment resistance in pancreas cancers. Similar properties may be operant in other solid tumors as well, so revisiting and characterizing fluid mechanics with modern techniques in other autochthonous cancers may be warranted.

  19. Sperm counts and serum follicle-stimulating hormone levels before and after radiotherapy and chemotherapy in men with testicular germ cell cancer

    SciTech Connect

    Berthelsen, J.G.

    1984-02-01

    Sperm counts were low (median, 15 X 10(6) per ejaculate) and serum follicle-stimulating hormone (FSH) levels were moderately elevated (median, 31 IU/l) after unilateral orchiectomy and immediately before radiotherapy and chemotherapy in 34 patients with seminomas and 20 patients with nonseminomatous germ cell tumors. The scattered radiation (0.2 to 1.3 Gray (Gy)) reaching the remaining testicle during radiotherapy caused azoospermia in more than two thirds of the patients. A median of 540 days elapsed after the end of treatment before spermatozoa were again found in semen samples, while a median of 1250 days passed before the pretreatment sperm count was reached. One to 5 years after treatment, sperm counts were still low (median, 6 X 10(6) per ejaculate) and serum FSH was elevated (median, 61 IU/l). The adjuvant chemotherapy given to the 20 patients with nonseminomatous tumors did not appear to affect restitution appreciably.

  20. 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

  1. Dendritic cell count in the graft predicts relapse in patients with hematologic malignancies undergoing an HLA-matched related allogeneic peripheral blood stem cell transplant.

    PubMed

    Rajasekar, Reena; Lakshmi, Kavitha M; George, Biju; Viswabandya, Auro; Thirugnanam, Rajasekar; Abraham, Aby; Chandy, Mammen; Srivastava, Alok; Mathews, Vikram

    2010-06-01

    We investigated the impact of the number of infused and reconstituted immunocompetent cells including dendritic cells (DCs) on clinical outcome of patients with hematologic malignancies undergoing an allogeneic peripheral blood stem cell transplantation. Sixty-nine consecutive patients with hematologic malignancies were included in the analysis. The median age of the cohort was 32 years (range: 2-62 years) and there were 39 (57%) males. Twenty-one (30%) patients relapsed with a cumulative incidence of 44 % +/- 14% at a median follow up of 28 months. On a multivariate analysis, a high plasmacytoid dendritic cell (PC) content in the graft was associated with higher risk of relapse. The patients were further categorized based on the median PC counts in the graft as high (> or =2.3 x 10(6)/kg) and low (<2.3 x 10(6)/kg) groups. The baseline characteristics of these 2 groups were comparable. The group that had a high PC content in the graft had significantly higher risk of relapse and lower overall survival (OS) and event-free survival (EFS). Our data suggests that PC content in the graft predicts clinical outcomes such as relapse and survival in patients with hematologic malignancies undergoing an allogeneic HLA matched related peripheral blood stem cell transplantation. There is potential for pretransplant manipulation of this cellular subset in the graft.

  2. 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

  3. Osmolarity regulates chondrogenic differentiation potential of synovial fluid derived mesenchymal progenitor cells.

    PubMed

    Bertram, Karri L; Krawetz, Roman J

    2012-06-08

    Cartilage is one of few tissues where adult stem/progenitor cells have not been putatively identified. Recent studies have provided strong evidence that a sub-population of mesenchymal progenitor cells (MPCs) derived from the synovial fluid may be able to affect some degree of cartilage repair both in vivo and in vitro/ex vivo, however this does not appear to be the case in patients with arthritis. Previously, it has been found that synovial fluid osmolarity is decreased in patients with osteoarthritis (OA) or Rheumatoid arthritis (RA) and these changes in osmolarity have been linked to changes in chondrocyte gene regulation. However, it is yet unknown if changes in osmolarity regulate the gene expression in synovial fluid MPCs (sfMPCs), and by extension, chondrogenesis of this cell population. In the present study we have collected synovial fluid samples from normal, OA and RA knee joints, quantified the osmolarity of the fluid and modified the culture/differentiation media to span a range of osmolarities (264-375 mOsm). Chondrogenesis was measured with Alcian blue staining of cultures in addition to quantitative PCR (qPCR) using probes to Sox9, ACAN and Col2A1. Overall, sfMPCs from arthritic joints demonstrated decreased chondrogenic potential compared to sfMPCs isolated from normal synovial fluid. Furthermore, the sfMPCs retained increased chondrogenic potential if differentiated under the same osmolarity conditions for which they were initially derived within. In conclusion, it does appear the synovial fluid osmolarity regulates the chondrogenic potential of sfMPCs, however, further study is required to elucidate the mechanism by which the changes in osmolarity are sensed by the cells and regulate chondrogenic gene expression.

  4. Fluorescent intensity-based differential counting of FITC-doped silica nanoparticles: applications of CD4+ T-cell detection in microchip-type flowcytometers

    NASA Astrophysics Data System (ADS)

    Yun, Hoyoung; Bang, Hyunwoo; Lee, Won Gu; Lim, Hyunchang; Park, Junha; Lee, Joonmo; Riaz, Asif; Cho, Keunchang; Chung, Chanil; Han, Dong-Chul; Chang, Jun Keun

    2007-12-01

    Although CD4+ T-cells are an important target of HIV detection, there have been still major problems in making a diagnosis and monitoring in the third world and the region with few medical facilities. Then, it is necessary to use portable diagnosis devices at low cost when you put an enumeration of CD4+ T-cells. In general, the counting of CD4 below 200cells/uL makes it necessary to initiate antiretroviral treatment in adults (over 13 years old). However, lymphocyte subsets (including CD4 counts) of infants and young children are higher than those of adults. This fact shows the percentage of CD4+ T-cells of blood subsets, i.e., CD4/CD45%, CD4/CD8% or CD4/CD3% means a more reliable indicator of HIV infection than absolute counts in children. To know the percentage of CD4+ T-cell by using two fluorescent dyes of different emission wavelength, at least, one laser and two PMT detectors are in general needed. Then, it is so hard to develop a portable device like a 'toaster size' because this makes such a device more complex including many peripheral modules. In this study, we developed a novel technique to control the intensity of fluorescent dye-doped silica nanoparticles. I synthesized FITC-doped silica nanoparticles conjugated CD4 antibody 10 times brighter than FITC-conjugated CD45 antibody. With the difference of intensity of two fluorescent dyes, we measured two parameters by using only a single detector and laser. Most experiments were achieved with uFACS (microfabricated fluorescence-activated cell sorter) on an inverted microscope (IX71, Olympus). In conclusion, this method enables us to discriminate the difference between CD4 and CD45 in an intensity domain simultaneously. Furthermore, this technique would make it possible develop much cheaper and smaller devices which can count the number of CD4 T-cells.

  5. Lamin A/C deficiency reduces circulating tumor cell resistance to fluid shear stress

    PubMed Central

    Denais, Celine; Chan, Maxine F.; Wang, Zhexiao; Lammerding, Jan

    2015-01-01

    Metastasis contributes to over 90% of cancer-related deaths and is initiated when cancer cells detach from the primary tumor, invade the basement membrane, and enter the circulation as circulating tumor cells (CTCs). While metastasis is viewed as an inefficient process with most CTCs dying within the bloodstream, it is evident that some CTCs are capable of resisting hemodynamic shear forces to form secondary tumors in distant tissues. We hypothesized that nuclear lamins A and C (A/C) act as key structural components within CTCs necessary to resist destruction from elevated shear forces of the bloodstream. Herein, we show that, compared with nonmalignant epithelial cells, tumor cells are resistant to elevated fluid shear forces in vitro that mimic those within the bloodstream, as evidenced by significant decreases in cellular apoptosis and necrosis. Knockdown of lamin A/C significantly reduced tumor cell resistance to fluid shear stress, with significantly increased cell death compared with parental tumor cell and nontargeting controls. Interestingly, lamin A/C knockdown increased shear stress-induced tumor cell apoptosis, but did not significantly affect cellular necrosis. These data demonstrate that lamin A/C is an important structural component that enables tumor cell resistance to fluid shear stress-mediated death in the bloodstream, and may thus facilitate survival and hematogenous metastasis of CTCs. PMID:26447202

  6. Computational fluid dynamic simulation of aggregation of deformable cells in a shear flow.

    PubMed

    Bagchi, Prosenjit; Johnson, Paul C; Popel, Aleksander S

    2005-12-01

    We present computational fluid dynamic (CFD) simulation of aggregation of two deformable cells in a shearflow. This work is motivated by an attempt to develop computational models of aggregation of red blood cells (RBCs). Aggregation of RBCs is a major determinant of blood viscosity in microcirculation under physiological and pathological conditions. Deformability of the RBCs plays a major role in determining their aggregability. Deformability depends on the viscosity of the cytoplasmic fluid and on the rigidity of the cell membrane, in a macroscopic sense. This paper presents a computational study of RBC aggregation that takes into account the rheology of the cells as well as cell-cell adhesion kinetics. The simulation technique considered here is two dimensional and based on the front tracking/immersed boundary method for multiple fluids. Results presented here are on the dynamic events of aggregate formation between two cells, and its subsequent motion, rolling, deformation, and breakage. We show that the rheological properties of the cells have significant effects on the dynamics of the aggregate. A stable aggregate is formed at higher cytoplasmic viscosity and membrane rigidity. We also show that the bonds formed between the cells change in a cyclic manner as the aggregate rolls in a shearflow. The cyclic behavior is related to the rolling orientation of the aggregate. The frequency and amplitude of oscillation in the number of bonds also depend on the rheological properties.

  7. Titanium dioxide nanoparticles induce cytotoxicity and reduce mitotic index in human amniotic fluid-derived cells.

    PubMed

    Acar, M S; Bulut, Z B; Ateş, A; Nami, B; Koçak, N; Yıldız, B

    2015-01-01

    Titanium dioxide (TiO2) nanoparticles (NPs) are commonly used materials present in many consumables for which most people are exposed to. The biological hazards of the NPs on human health have been demonstrated previously. In this study, we aimed to assess the cytotoxicity potency of TiO2 NPs on the primary human amniotic fluid cells. The cells derived from amniotic fluid were treated with different dosages of TiO2 NPs for some periods. Cell adhesion status was assessed using a light microscopic observation. Cell proliferation and cell death rates were determined using trypan blue staining and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. Also, mitotic index was determined using fluorescence in situ hybridization with chromosome 8 centromer-specific DNA probe. Disrupted cell adhesion, decreased proliferation, and increased mortality rates were detected in the cells that were treated with TiO2 NPs depending on the dosage (p < 0.001). Also, reduced mitotic index was determined in the cells depending on the time and TiO2 dosage when compared with the controls (p < 0.0001). These results showed that TiO2 NPs have high cytotoxicity for amniotic fluid-derived cells. Therefore, different products containing TiO2 NPs should be used with care, especially for pregnant women.

  8. Adipose tissue-derived mesenchymal stem cells acquire bone cell-like responsiveness to fluid shear stress on osteogenic stimulation.

    PubMed

    Knippenberg, Marlene; Helder, Marco N; Doulabi, Behrouz Zandieh; Semeins, Cornelis M; Wuisman, Paul I J M; Klein-Nulend, Jenneke

    2005-01-01

    To engineer bone tissue, mechanosensitive cells are needed that are able to perform bone cell-specific functions, such as (re)modeling of bone tissue. In vivo, local bone mass and architecture are affected by mechanical loading, which is thought to provoke a cellular response via loading-induced flow of interstitial fluid. Adipose tissue is an easily accessible source of mesenchymal stem cells for bone tissue engineering, and is available in abundant amounts compared with bone marrow. We studied whether adipose tissue-derived mesenchymal stem cells (AT-MSCs) are responsive to mechanical loading by pulsating fluid flow (PFF) on osteogenic stimulation in vitro. We found that ATMSCs show a bone cell-like response to fluid shear stress as a result of PFF after the stimulation of osteogenic differentiation by 1,25-dihydroxyvitamin D3. PFF increased nitric oxide production, as well as upregulated cyclooxygenase-2, but not cyclooxygenase-1, gene expression in osteogenically stimulated AT-MSCs. These data suggest that AT-MSCs acquire bone cell-like responsiveness to pulsating fluid shear stress on 1,25-dihydroxyvitamin D3-induced osteogenic differentiation. ATMSCs might be able to perform bone cell-specific functions during bone (re)modeling in vivo and, therefore, provide a promising new tool for bone tissue engineering.

  9. Amniotic fluid derived stem cells give rise to neuron-like cells without a further differentiation potential into retina-like cells.

    PubMed

    Hartmann, K; Raabe, O; Wenisch, S; Arnhold, S

    2013-01-01

    Amniotic fluid contains heterogeneous cell types and has become an interesting source for obtaining fetal stem cells. These stem cells have a high proliferative capacity and a good differentiation potential and may thus be suitable for regenerative medicine. As there is increasing evidence, that these stem cells are also able to be directed into the neural lineage, in our study we investigated the neuronal and glial differentiation potential of these cells, so that they may also be applied to cure degenerative diseases of the retina. Mesenchymal stem cells were isolated from routine prenatal amniocentesis at 15 to 18 weeks of pregnancy of human amniotic fluid and expanded in the cell culture. Cells were cultivated according to standard procedures for mesenchymal stem cells and were differentiated along the neural lineage using various protocols. Furthermore, it was also tried to direct them into cell types of the retina as well as into endothelial cells. Cells of more than 72 amniotic fluid samples were collected and characterized. While after induction neural-like phenotypes could actually be detected, which was confirmed using neural marker proteins such as GFAP and ßIII tubulina further differentiation into retinal like cells could not reliably be shown. These data suggest that amniotic fluid derived cells are an interesting cell source, which may also give rise to neural-like cells. However, a more specific differentiation into neuronal and glial cells could not unequivocally be shown, so that further investigations have to becarried out.

  10. Design and fabrication of a magnetic fluid micropump for applications in direct methanol fuel cells

    NASA Astrophysics Data System (ADS)

    Lee, Shi-Min; Kuan, Yean-Der; Sung, Min-Feng

    Direct methanol fuel cells (DMFCs) are widely considered to have great potential for portable electric applications, and the power requirements for many of them are only a few watts. Therefore, a low power liquid pump is especially desirable for driving the methanol solution fuel for an active direct methanol fuel. The main objective of this paper is to design and fabricate a magnetic fluid micropump that has characteristics of low operation voltage and current and is suitable for use in DMFCs. Two prototypes were developed and tested. The magnetic fluid micropumps are successfully applied to drive the fuel to a DMFC, and measurements of the cell performance are also conducted.

  11. Characteristics of human amniotic fluid mesenchymal stem cells and their tropism to human ovarian cancer.

    PubMed

    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.

  12. Modeling CD4+ cell count increase over a six-year period in HIV-1-infected patients on highly active antiretroviral therapy in Senegal.

    PubMed

    De Beaudrap, Pierre; Etard, Jean-François; Diouf, Assane; Ndiaye, Ibrahima; Guèye, Ndèye Fatou; Guèye, Pape Mandoumbé; Sow, Papa Salif; Mboup, Souleymane; Ndoye, Ibra; Ecochard, René; Eric, Delaporte

    2009-06-01

    To assess the extents and determinants of long-term CD4 cell increases after initiation of antiretroviral therapy (ART), changes in CD4 cell counts were analyzed in a cohort of HIV-1-infected Senegalese using a mixed-effects model. After a median follow-up of 54 months, an average of 483 CD4 cells/mm3 (95% confidence interval [CI] = 331; 680) was reached. The average asymptote level was approximately 421 cells/mm3 (95% CI = 390; 454) in patients with < 200 cells/mm3 at baseline and approximately 500 cells/mm3 in patients with > 200 cells/mm3. The independent predictors of long-term CD4 cell reconstitution were the baseline CD4 cell count and the monthly average viral load over the entire follow-up. This good long-term immune reconstitution, optimal in subjects with low average viral loads and > 200 CD4 cells/mm3 at baseline, argues in favor of the earliest possible access to ART and underlines the importance of strict compliance with the treatment.

  13. Economic consequences of mastitis and withdrawal of milk with high somatic cell count in Swedish dairy herds.

    PubMed

    Nielsen, C; Ostergaard, S; Emanuelson, U; Andersson, H; Berglund, B; Strandberg, E

    2010-10-01

    The main aim was to assess the impact of mastitis on technical and economic results of a dairy herd under current Swedish farming conditions. The second aim was to investigate the effects obtained by withdrawing milk with high somatic cell count (SCC). A dynamic and stochastic simulation model, SimHerd, was used to study the effects of mastitis in a herd with 150 cows. Results given the initial incidence of mastitis (32 and 33 clinical and subclinical cases per 100 cow-years, respectively) were studied, together with the consequences of reducing or increasing the incidence of mastitis by 50%, modelling no clinical mastitis (CM) while keeping the incidence of subclinical mastitis (SCM) constant and vice versa. Six different strategies to withdraw milk with high SCC were compared. The decision to withdraw milk was based on herd-level information in three scenarios: withdrawal was initiated when the predicted bulk tank SCC exceeded 220 000, 200 000 or 180 000 cells/ml, and on cow-level information in three scenarios: withdrawal was initiated when the predicted SCC in an individual cow's milk exceeded 1 000 000, 750 000 or 500 000 cells/ml. The accuracy with which SCC was measured and predicted was assumed to affect the profitability of withdrawing milk with high SCC and this was investigated by applying high, low or no uncertainty to true SCC. The yearly avoidable cost of mastitis was estimated at €8235, assuming that the initial incidence of mastitis could be reduced by 50%. This cost corresponded to 5% of the herd net return given the initial incidence of mastitis. Expressed per cow-year, the avoidable cost of mastitis was €55. The costs per case of CM and SCM were estimated at €278 and €60, respectively. Withdrawing milk with high SCC was never profitable because this generated a substantial amount of milk withdrawal that was not offset by a sufficient increase in the average price per delivered kg milk. It had the most negative impact on net return when

  14. Herd management and social variables associated with bulk tank somatic cell count in dairy herds in the eastern United States.

    PubMed

    Schewe, R L; Kayitsinga, J; Contreras, G A; Odom, C; Coats, W A; Durst, P; Hovingh, E P; Martinez, R O; Mobley, R; Moore, S; Erskine, R J

    2015-11-01

    The ability to reduce somatic cell counts (SCC) and improve milk quality depends on the effective and consistent application of established mastitis control practices. The US dairy industry continues to rely more on nonfamily labor to perform critical tasks to maintain milk quality. Thus, it is important to understand dairy producer attitudes and beliefs relative to management practices, as well as employee performance, to advance milk quality within the changing structure of the dairy industry. To assess the adoption rate of mastitis control practices in United States dairy herds, as well as assess social variables, including attitudes toward employees relative to mastitis control, a survey was sent to 1,700 dairy farms in Michigan, Pennsylvania, and Florida in January and February of 2013. The survey included questions related to 7 major areas: sociodemographics and farm characteristics, milking proficiency, milking systems, cow environment, infected cow monitoring and treatment, farm labor, and attitudes toward mastitis and related antimicrobial use. The overall response rate was 41% (21% in Florida, 39% in Michigan, and 45% in Pennsylvania). Herd size ranged from 9 to 5,800 cows. Self-reported 3-mo geometric mean bulk tank SCC (BTSCC) for all states was 194,000 cells/mL. Multivariate analysis determined that proven mastitis control practices such as the use of internal teat sealants and blanket dry cow therapy, and not using water during udder preparation before milking, were associated with lower BTSCC. Additionally, farmer and manager beliefs and attitudes, including the perception of mastitis problems and the threshold of concern if BTSCC is above 300,000 cells/mL, were associated with BTSCC. Ensuring strict compliance with milking protocols, giving employees a financial or other penalty if BTSCC increased, and a perceived importance of reducing labor costs were negatively associated with BTSCC in farms with nonfamily employees. These findings highlight the

  15. 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/μl). We determined HIVp24-specific T-cell responses using standard Interferon-gamma (IFNγ) 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 IFNγ-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

  16. Obstructive renal injury: from fluid mechanics to molecular cell biology.

    PubMed

    Ucero, Alvaro C; Gonçalves, Sara; Benito-Martin, Alberto; Santamaría, Beatriz; Ramos, Adrian M; Berzal, Sergio; Ruiz-Ortega, Marta; Egido, Jesus; Ortiz, Alberto

    2010-04-22

    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-β1 (TGF-β1) 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.

  17. 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-β1 (TGF-β1) 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

  18. Potential role of culture mediums for successful isolation and neuronal differentiation of amniotic fluid stem cells.

    PubMed

    Orciani, M; Emanuelli, M; Martino, C; Pugnaloni, A; Tranquilli, A L; Di Primio, R

    2008-01-01

    In recent years, the use of stem cells has generated increasing interest in regenerative medicine and cancer therapies. The most potent stem cells derive from the inner cell mass during embryonic development and their use yields serious ethical and methodological problems. Recently, a number of reports suggests that another suitable source of multipotent stem cells may be the amniotic fluid. Amniotic fluid mesenchymal stem cells (AFMSCs) are capable of extensive self-renewal, able to differentiate in specialized cells representative of all three germ layers, do not show ethical restriction, and display minimal risks of teratomas and a very low immunogenity. For all these reasons, amniotic fluid appears as a promising alternative source for stem cell therapy. Their recent discovery implies a lack of knowledge of their specific features as well as the existence of a protocol universally recognized as the most suitable for their isolation, growth and long-term conservation. In this study, we isolated stem cells from six amniotic fluids; these cells were cultured with three different culture mediums (Mesenchymal Stem Cell Medium (MSCGM), PC-1 and RPMI-1640), characterized by cytofluorimetric analysis, and then either frozen or induced to neuronal differentiation. Even if the immunophenotype seemed not to be influenced by culture medium (all six samples cultured in the above-mentioned mediums expressed surface antigens commonly found on stem cells), cells showed different abilities to differentiate into neuron-like cells and to re-start the culture after short/long-term storage. Cells isolated and cultured in MSCGM showed the highest proliferation rate, and formed neuron-like cells when sub-plated with neuronal differentiation medium. Cells from PC-1, on the contrary, displayed an increased ability to re-start culture after short/long term storage. Finally, cells from RPMI-1640, even if expressing stem cells markers, were not able to differentiate in neuron-like cells

  19. The diagnostic value of white cell count, C-reactive protein and bilirubin in acute appendicitis and its complications

    PubMed Central

    Parashar, D; Lin, R; Antonowicz, S; Wells, AD; Bajwa, FM; Krijgsman, B

    2013-01-01

    Introduction Inflammatory markers such as white cell count (WCC) and C-reactive protein (CRP) and, more recently, bilirubin have been used as adjuncts in the diagnosis of appendicitis. The aim of this study was to determine the diagnostic accuracy of the above markers in acute and perforated appendicitis as well as their value in excluding the condition. Methods A retrospective analysis of 1,169 appendicectomies was performed. Patients were grouped according to histological examination of appendicectomy specimens (normal appendix = NA, acute appendicitis = AA, perforated appendicitis = PA) and preoperative laboratory test results were correlated. Receiver operating characteristic (ROC) curve area analysis (area under the curve [AUC]) was performed to examine diagnostic accuracy. Results ROC analysis of all laboratory variables showed that no independent variable was diagnostic for AA. Good diagnostic accuracy was seen for AA when all variables were combined (WCC/CRP/bilirubin combined AUC: 0.8173). In PA, the median CRP level was significantly higher than that of AA (158mg/l vs 30mg, p<0.0001). CRP also showed the highest sensitivity (100%) and negative predictive value (100%) for PA. CRP had the highest diagnostic accuracy in PA (AUC: 0.9322) and this was increased when it was combined with WCC (AUC: 0.9388). Bilirubin added no diagnostic value in PA. Normal levels of WCC, CRP and bilirubin could not rule out appendicitis. Conclusions CRP provides the highest diagnostic accuracy for PA. Bilirubin did not provide any discriminatory value for AA and its complications. Normal inflammatory markers cannot exclude appendicitis, which remains a clinical diagnosis. PMID:23827295

  20. Changes in B-Cell Counts and Percentages during Primary HIV Infection Associated with Disease Progression in HIV-Infected Men Who Have Sex with Men: A Preliminary Study

    PubMed Central

    Cui, Chen; Jiang, Yongjun; Zhang, Zining; Hu, Qinghai; Chu, Zhenxing; Xu, Junjie; Zhao, Bin; Ding, Haibo; Liu, Jing; Han, Xiaoxu; Cao, Yaming; Shang, Hong

    2015-01-01

    Numerous anomalies in B-cell phenotypes and functions have been described in HIV-infected individuals. However, the actual relationship between B cells and disease progression remains unclear. In this study, we investigated B-cell counts/percentages during a 12-month infection period in HIV-infected individuals that eventually developed into typical progressors (TPs) or rapid progressors (RPs). We found, after 12 months of infection, the baseline B-cell counts/percentages correlated positively with CD4+ T-cell counts (P = 0.0006 and P = 0.026) and negatively with HIV viral set points (P = 0.014 and P = 0.002). Kaplan-Meier survival analysis showed that high baseline B-cell counts/percentages were associated with a slow CD4-cell decline. B-cell kinetics indicated the baseline B-cell counts/percentages could be factors distinguishing between TPs and RPs. The combination of the baseline B-cell counts and percentages was associated with rapid disease progression (a 80.7% predictive value as measured by the area under the curve). These results indicate that the baseline B-cell counts/percentages might be associated with HIV disease progression. PMID:26436092

  1. Influence of birth weight on white blood cell count in biracial (black-white) children, adolescents, and young adults: the Bogalusa Heart Study.

    PubMed

    Chen, Wei; Srinivasan, Sathanur R; Berenson, Gerald S

    2009-01-15

    The effect of birth weight on white blood cell (WBC) count among blacks and whites was examined in 2,080 children (aged 4-11 years, 57.4% white, and 49.2% male), 892 adolescents (aged 12-17 years, 57.2% white, and 50.8% male), and 1,872 adults (aged 18-38 years, 68.4% white, and 41.9% male) from Bogalusa, Louisiana, in 2005. After adjustment for age, sex, race, body mass index, and smoking status (in adolescents and adults), the WBC count decreased across quartiles of increasing birth weight specific for race, sex, and gestational age in children (P(trend) = 0.0007) and adults (P(trend) = 0.005). In multivariate regression analyses that included the covariates above, birth weight was inversely associated with WBC count in children (beta coefficients (unit, cells/microL per kg) = -256, -241, and -251 for whites, blacks, and the combined sample, with P = 0.003, 0.029, and <0.001, respectively) and in adults (beta = -224 and -211 for whites and the combined sample, with P = 0.015 and 0.008, respectively). These results show that low birth weight is associated with increased systemic inflammation as depicted by the WBC count in childhood and adulthood, thereby potentially linking fetal growth retardation to cardiovascular disease and diabetes.

  2. Low-Cost Disposable Cartridge for Performing a White Blood Cell Count and Partial Differential at the Point-of-Care.

    PubMed

    Majors, Catherine E; Pawlowski, Michal E; Tkaczyk, Tomasz; Richards-Kortum, Rebecca R

    2014-10-01

    Being able to perform a white blood cell (WBC) count and differential is a crucial laboratory test for basic diagnostic practices. In this paper, we demonstrate proof of concept results for a disposable cartridge that could be used to perform a WBC count and 3-part differential at the point-of-care. The cartridge is composed of a glass slide, a layer of transfer tape, and a glass cover slip and incorporates acridine orange for cell staining and sub-type differentiation; the stained blood is then imaged, and image analysis techniques return a WBC count and 3-part differential. The cartridge was tested on a laboratory microscope with 3 normal samples, and had promising results with 85.7% of images resulting in a WBC count with ±15% of the true value. Further, the 3-part differential concentrations determined using the disposable cartridge had strong correlations with the true concentrations (R(2) values of 0.9986, 0.9421, and 0.6942 for granulocytes, lymphocytes, and monocytes, respectively). Preliminary designs for a low-cost, portable microscope have been created and are currently being prototyped.

  3. Abnormal Glucose Tolerance, White Blood Cell Count, and Telomere Length in Newly Diagnosed, Antidepressant-Naïve Patients with Depression

    PubMed Central

    Garcia-Rizo, Clemente; Fernandez-Egea, Emilio; Miller, Brian J.; Oliveira, Cristina; Justicia, Azucena; Griffith, Jeffrey K.; Heaphy, Christopher M.; Bernardo, Miguel; Kirkpatrick, Brian

    2012-01-01

    Chronic mood disorders have been associated with a shortened telomere, a marker of increased mortality rate and ageing, and impaired cellular immunity. However, treatment may confound these relationships. We examined the relationship of glucose tolerance, white blood cell count and telomere length to depression in newly diagnosed, antidepressant-naïve patients. Subjects with major depression (n=15), and matched healthy control subjects (n=70) underwent a two-hour oral glucose tolerance test and evaluation of blood cell count and telomere content. The depression group had significantly higher two-hour glucose concentrations and a lower lymphocyte count than control subjects (respective means [SD] for two-hour glucose were 125.0 mg/dL [67.9] vs 84.6 [25.6] (p<.001); for lymphocyte count 2.1 × 109/L [0.6] vs. 2.5 ×109/L [0.7] p=.028).Telomere content was significantly shortened in the depression group (87.9 [7.6]) compared to control subjects (101.0 [14.3]; p<0.01). Abnormal glucose tolerance, lymphopenia and a shortened telomere are present early in the course of depression independently of the confounding effect of antidepressant treatment, supporting the concept of major depression as an accelerated ageing disease. PMID:23207109

  4. Low-Cost Disposable Cartridge for Performing a White Blood Cell Count and Partial Differential at the Point-of-Care*

    PubMed Central

    Majors, Catherine E.; Pawlowski, Michal E.; Tkaczyk, Tomasz; Richards-Kortum, Rebecca R.

    2015-01-01

    Being able to perform a white blood cell (WBC) count and differential is a crucial laboratory test for basic diagnostic practices. In this paper, we demonstrate proof of concept results for a disposable cartridge that could be used to perform a WBC count and 3-part differential at the point-of-care. The cartridge is composed of a glass slide, a layer of transfer tape, and a glass cover slip and incorporates acridine orange for cell staining and sub-type differentiation; the stained blood is then imaged, and image analysis techniques return a WBC count and 3-part differential. The cartridge was tested on a laboratory microscope with 3 normal samples, and had promising results with 85.7% of images resulting in a WBC count with ±15% of the true value. Further, the 3-part differential concentrations determined using the disposable cartridge had strong correlations with the true concentrations (R2 values of 0.9986, 0.9421, and 0.6942 for granulocytes, lymphocytes, and monocytes, respectively). Preliminary designs for a low-cost, portable microscope have been created and are currently being prototyped. PMID:25918749

  5. Periovulatory follicular fluid levels of estradiol trigger inflammatory and DNA damage responses in oviduct epithelial cells

    PubMed Central

    Palma-Vera, Sergio E.; Schoen, Jennifer; Chen, Shuai

    2017-01-01

    Objective Ovarian steroid hormones (mainly E2 and P4) regulate oviduct physiology. Serum-E2 acts on the oviduct epithelium from the basolateral cell compartment. Upon ovulation, the apical compartment of the oviduct epithelium is temporarily exposed to follicular fluid, which contains much higher levels of E2 than serum. The aim of this study was to evaluate the effects of human periovulatory follicular fluid levels of E2 on oviduct epithelial cells using two porcine in vitro models. Methods A cell line derived from the porcine oviductal epithelium (CCLV-RIE270) was characterized (lineage markers, proliferation characteristics and transformation status). Primary porcine oviduct epithelial cells (POEC) were cultured in air-liquid interface and differentiation was assessed histologically. Both cultures were exposed to E2 (10 ng/ml and 200 ng/ml). Proliferation of CCLV-RIE270 and POEC was determined by real-time impedance monitoring and immunohistochemical detection of Ki67. Furthermore, marker gene expression for DNA damage response (DDR) and inflammation was quantified. Results CCLV-RIE270 was not transformed and exhibited properties of secretory oviduct epithelial cells. Periovulatory follicular fluid levels of E2 (200 ng/ml) upregulated the expression of inflammatory genes in CCLV-RIE270 but not in POEC (except for IL8). Expression of DDR genes was elevated in both models. A significant increase in cell proliferation could not be detected in response to E2. Conclusions CCLV-RIE270 and POEC are complementary models to evaluate the consequences of oviduct exposure to follicular fluid components. Single administration of periovulatory follicular fluid E2 levels trigger inflammatory and DNA damage responses, but not proliferation in oviduct epithelial cells. PMID:28231273

  6. Simulation of fluid-solid coexistence via thermodynamic integration using a modified cell model.

    PubMed

    Nayhouse, Michael; Amlani, Ankur M; Heng, Vincent R; Orkoulas, G

    2012-04-18

    Despite recent advances, precise simulation of fluid-solid transitions still remains a challenging task. Thermodynamic integration techniques are the simplest methods to study fluid-solid coexistence. These methods are based on the calculation of the free energies of the fluid and the solid phases, starting from a state of known free energy which is usually an ideal-gas state. Despite their simplicity, the main drawback of thermodynamic integration techniques is the large number of states that must be simulated. In the present work, a thermodynamic integration technique, which reduces the number of simulated states, is proposed and tested on a system of particles interacting via an inverse twelfth-power potential energy function. The simulations are implemented at constant pressure and the solid phase is modeled according to the constrained cell model of Hoover and Ree. The fluid and the solid phases are linked together by performing constant-pressure simulations of a modified cell model. The modified cell model, which was originally proposed by Hoover and Ree, facilitates transitions between the fluid and the solid phase by tuning a homogeneous external field. This model is simulated on a constant-pressure path for a series of progressively increasing values of the field, thus allowing for direct determination of the free energy difference between the fluid and the solid phase via histogram reweighting. The size-dependent results are analyzed using histogram reweighting and finite-size scaling techniques. The scaling analysis is based on studying the size-dependent behavior of the second- and higher-order derivatives of the free energy as well as the dimensionless moment ratios of the order parameter. The results clearly demonstrate the importance of accounting for size effects in simulation studies of fluid-solid transitions.

  7. Spinal fluid lymphocytes responsive to autologous and allogeneic cells in multiple sclerosis and control individuals.

    PubMed Central

    Birnbaum, G; Kotilinek, L; Schwartz, M; Sternad, M

    1984-01-01

    Spinal fluid lymphocytes from multiple sclerosis (MS) patients and controls were stimulated with either autologous non-T cells or with allogeneic non-T cells followed by stimulation with autologous non-T lymphocytes. Cells responding to these stimuli were cloned and their proliferative responses to autologous and allogeneic MS and normal non-T cells were measured. Large numbers of clones with specific patterns of reaction to both autologous and allogeneic cells were obtained from lymphocytes in MS cerebrospinal fluid (CSF), but only occasionally from cells in control CSF. Patterns of responses among clones from a particular CSF were similar and often identical, which suggested that cells in MS CSF were relatively restricted in their specificities. Surface antigen phenotyping of the clones showed them to be predominantly OKT4+, with 13% OKT8+ and 11% OKT4+8+. Peripheral T cells that were stimulated and cultured in parallel with CSF cells were different in that they usually did not give rise to as many clones nor were their patterns of response similar. Many CSF clones were heteroclitic, that is they responded to particular allogeneic cells but not autologous cells. Lymphocytes in MS CSF thus appear to represent a selected population of cells with a high frequency of responsiveness to autologous and allogeneic antigens. Such responses may be evidence for immune regulation within the central nervous system or could represent responses to altered-self antigens. PMID:6237121

  8. Synovial fluid and synovial membrane mesenchymal stem cells: latest discoveries and therapeutic perspectives.

    PubMed

    de Sousa, Eduardo Branco; Casado, Priscila Ladeira; Moura Neto, Vivaldo; Duarte, Maria Eugenia Leite; Aguiar, Diego Pinheiro

    2014-10-03

    Mesenchymal stem cells (MSCs) have the ability to differentiate into osteoblasts, chondroblasts, adipocytes, and even myoblasts. Most studies have focused on finding MSCs in different parts of the body for medical treatment. Every joint structure, including bone, joint fat, articular cartilage, and synovium, potentially contains resident MSCs. Recently, a progenitor cell population has been found in synovial fluid and showed similarities with both bone marrow and synovial membrane MSCs. Synovial fluid MSCs have been studied in healthy persons and osteoarthritic patients in order to explore its potential for treatment of some orthopedic disorders. Here, we briefly review the current knowledge on synovial fluid MSCs, their origin, relation to some orthopedic diseases, and future applications.

  9. Basal CD34+ Cell Count Predicts Peripheral Blood Stem Cell Mobilization in Healthy Donors after Administration of G-CSF: a Longitudinal, Prospective, Observational, Single-Center, Cohort Study.

    PubMed

    Martino, Massimo; Gori, Mercedes; Pitino, Annalisa; Gentile, Massimo; Dattola, Antonia; Pontari, Antonella; Vigna, Ernesto; Moscato, Tiziana; Recchia, Anna Grazia; Barilla', Santina; Tripepi, Giovanni; Morabito, Fortunato

    2017-03-24

    A longitudinal, prospective, observational, single-center, cohort study on healthy donors (HDs) was designed to identify predictors of CD34+ cells on day 5 with emphasis on the predictive value of the basal CD34+ cell count. As potential predictors of mobilization, age, sex, body weight, height, blood volume as well as white blood cell count, peripheral blood (PB) mononuclear cells, platelet count, hematocrit, and hemoglobin levels were considered. Two different evaluations of CD34+ cell counts were determined for each donor: baseline (before granulocyte colony-stimulating factor [G-CSF] administration) and in PB after G-CSF administration on the morning of the fifth day (day 5). A total 128 consecutive HDs (66 males) with a median age of 43 years were enrolled. CD34+ levels on day 5 displayed a non-normal distribution with a median value of 75.5 cells/µL. To account for the non-normal distribution of the dependent variable, a quantile regression analysis to predict CD34+ on day 5 using the baseline value of CD34+ as the key predictor was performed. On crude analysis, a baseline value of CD34+ ranging from 0.5 to 1 cells/µL predicts a median value of 50 cells/µL on day 5; a value of 2 cells/µL predicts a median value of 70.7 cells/µL; a value of 3-4 cells/µL predicts a median value of 91.3 cells/µL, and a value ≥ 5 predicts a median value of 112 cells/µL. In conclusion, the baseline PB CD34+ cell count correlates with the effectiveness of allogeneic PBSC mobilization and could be useful to plan the collection.

  10. 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.

  11. Characteristics of long-term asymptomatic infection with human immunodeficiency virus type 1 in men with normal and low CD4+ cell counts.

    PubMed

    Keet, I P; Krol, A; Klein, M R; Veugelers, P; de Wit, J; Roos, M; Koot, M; Goudsmit, J; Miedema, F; Coutinho, R A

    1994-06-01

    From a cohort study of homosexual men in Amsterdam, 61 human immunodeficiency virus (HIV)-infected men who had remained asymptomatic for at least 7 years were identified. In a nested case control study, these men were compared with 142 men who progressed symptomatic HIV infection (CDC class IV) within 7 years, regarding laboratory markers, sexual behavior, psychologic coping, and drug use. Of the 61 long-term asymptomatic men, 13 had a CD4+ cell count > or = 500/mm3 after 7 years; in 2 of these 13, the CD4+ cell count had not declined during follow-up. Independent of CD4+ cell count, long-term asymptomatic HIV-1 infection was characterized by stable T cell reactivity after stimulation with monoclonal CD3 antibodies, seropositivity for antibodies to HIV core proteins, and the absence of hepatitis B markers. No association with markers of high-risk sex or the recreational use of drugs was found. Long-term asymptomatic men had a slightly lower score regarding the coping behavior active problem-solving; no other associations with coping behaviors were found.

  12. [Pilot study of the Deosan-RMTK (rapid mastitis test kit), a diagnostic test for the detection of cows with high cell count].

    PubMed

    Noordhuizen, J P; Stassen, E N; Klerx, I

    1993-05-15

    The Deosan-Rapid Mastitis Test Kit (RMTK) was evaluated in 226 lactating dairy cows on 6 farms. The Fossomatic method was used as reference standard for somatic cell counts in cows milk. The RMTK test-principle regards the reaction of the enzyme catalase released from cells in milk with the H2O2 in the RMTK-reagent. For the threshold cell count values of 250,000, 400,000 and 800,000/ml the following 95% confidence intervals were found: sensitivity 0.60-0.99 specificity 0.42-0.83, predictive value positive 0.22-0.46, predictive value negative 0.84-0.99 and kappa-value 0.14-0.52. Because this test will be most useful when the positive predictive value would be high, it is concluded that the RMTK in this study population was not an adequate tool for the detection of cows with somatic cell counts over 250,000/ml milk.

  13. Quarter and cow risk factors associated with a somatic cell count greater than 199,000 cells per milliliter in united Kingdom dairy cows

    PubMed Central

    Breen, J. E.; Bradley, A. J.; Green, M. J.

    2009-01-01

    Quarter and cow risk factors associated with a somatic cell count (SCC) >199,000 cells/mL at the next milk recording during lactation were investigated during a 12-mo longitudinal study on 8 commercial Holstein-Friesian dairy herds in Southwest England, United Kingdom. The individual risk factors studied on 1,677 cows included assessments of udder and leg hygiene, teat-end callosity and hyperkeratosis, body condition score (BCS), and measurements of monthly milk quality and yield. The outcome variable used for statistical analysis was the next recorded individual cow SCC >199,000 cells/mL. Statistical analysis included use of generalized linear mixed models. Significant covariates associated with an increased risk of SCC >199,000 cells/mL were increasing parity, increasing month of lactation, previous SCC (SCC 200,000 cells/mL and greater, odds ratio = 7.12), and cows with a BCS <1.5 (odds ratio = 2.09) or BCS >3.5 (odds ratio = 2.20). Significant covariates associated with a reduced risk of SCC >199,000 cells/mL were cows with contamination of the skin of the udder and quarters with mild (odds ratio = 0.65) and moderate (odds ratio = 0.62) hyperkeratosis of the teat-end. These results suggest that individual quarter and cow-level factors are important in the acquisition of intramammary infections as measured by SCC during lactation. Cow energy status, as measured by BCS, may influence the risk of intramammary infection during lactation. PMID:19528588

  14. Amniotic Fluid Stem Cells: A Novel Source for Modeling of Human Genetic Diseases.

    PubMed

    Antonucci, Ivana; Provenzano, Martina; Rodrigues, Melissa; Pantalone, Andrea; Salini, Vincenzo; Ballerini, Patrizia; Borlongan, Cesar V; Stuppia, Liborio

    2016-04-22

    In recent years, great interest has been devoted to the use of Induced Pluripotent Stem cells (iPS) for modeling of human genetic diseases, due to the possibility of reprogramming somatic cells of affected patients into pluripotent cells, enabling differentiation into several cell types, and allowing investigations into the molecular mechanisms of the disease. However, the protocol of iPS generation still suffers from technical limitations, showing low efficiency, being expensive and time consuming. Amniotic Fluid Stem cells (AFS) represent a potential alternative novel source of stem cells for modeling of human genetic diseases. In fact, by means of prenatal diagnosis, a number of fetuses affected by chromosomal or Mendelian diseases can be identified, and the amniotic fluid collected for genetic testing can be used, after diagnosis, for the isolation, culture and differentiation of AFS cells. This can provide a useful stem cell model for the investigation of the molecular basis of the diagnosed disease without the necessity of producing iPS, since AFS cells show some features of pluripotency and are able to differentiate in cells derived from all three germ layers "in vitro". In this article, we describe the potential benefits provided by using AFS cells in the modeling of human genetic diseases.

  15. Amniotic Fluid Stem Cells: A Novel Source for Modeling of Human Genetic Diseases

    PubMed Central

    Antonucci, Ivana; Provenzano, Martina; Rodrigues, Melissa; Pantalone, Andrea; Salini, Vincenzo; Ballerini, Patrizia; Borlongan, Cesar V.; Stuppia, Liborio

    2016-01-01

    In recent years, great interest has been devoted to the use of Induced Pluripotent Stem cells (iPS) for modeling of human genetic diseases, due to the possibility of reprogramming somatic cells of affected patients into pluripotent cells, enabling differentiation into several cell types, and allowing investigations into the molecular mechanisms of the disease. However, the protocol of iPS generation still suffers from technical limitations, showing low efficiency, being expensive and time consuming. Amniotic Fluid Stem cells (AFS) represent a potential alternative novel source of stem cells for modeling of human genetic diseases. In fact, by means of prenatal diagnosis, a number of fetuses affected by chromosomal or Mendelian diseases can be identified, and the amniotic fluid collected for genetic testing can be used, after diagnosis, for the isolation, culture and differentiation of AFS cells. This can provide a useful stem cell model for the investigation of the molecular basis of the diagnosed disease without the necessity of producing iPS, since AFS cells show some features of pluripotency and are able to differentiate in cells derived from all three germ layers “in vitro”. In this article, we describe the potential benefits provided by using AFS cells in the modeling of human genetic diseases. PMID:27110774

  16. Multiphase modelling of the effect of fluid shear stress on cell yield and distribution in a hollow fibre membrane bioreactor.

    PubMed

    Pearson, Natalie C; Waters, Sarah L; Oliver, James M; Shipley, Rebecca J

    2015-04-01

    We present a simplified two-dimensional model of fluid flow, nutrient transport and cell distribution in a hollow fibre membrane bioreactor, with the aim of exploring how fluid flow can be used to control the distribution and yield of a cell population which is sensitive to both fluid shear stress and nutrient concentration. The cells are seeded in a scaffold in a layer on top of the hollow fibre, only partially occupying the extracapillary space. Above this layer is a region of free-flowing fluid which we refer to as the upper fluid layer. The flow in the lumen and upper fluid layer is described by the Stokes equations, whilst the flow in the porous fibre membrane is assumed to follow Darcy's law. Porous mixture theory is used to model the dynamics of and interactions between the cells, scaffold and fluid in the cell-scaffold construct. The concentration of a limiting nutrient (e.g. oxygen) is governed by an advection-reaction-diffusion equation in each region. Through exploitation of the small aspect ratio of each region and asymptotic analysis, we derive a coupled system of partial differential equations for the cell volume fraction and nutrient concentration. We use this model to investigate the effect of mechanotransduction on the distribution and yield of the cell population, by considering cases in which cell proliferation is either enhanced or limited by fluid shear stress and by varying experimentally controllable parameters such as flow rate and cell-scaffold construct thickness.

  17. A new flexible titanium foil cell for hydrothermal experiments and fluid sampling.

    PubMed

    Wu, Shi-Jun; Cai, Min-Jian; Yang, Can-Jun; Li, Ke-Wei

    2016-09-01

    This paper describes the design of a flexible titanium foil cell, as well as its applications in hydrothermal experiments and in non-contaminating storage of seafloor hydrothermal fluids. A flexible cell constructed totally from pure titanium (Grade 1) can be used in corrosive environment because of the excellent chemical stability and temperature tolerance of the material. Theoretical calculation and finite element analysis of the titanium foil cell have been conducted to identify its flexibility and deformation mode. Two applications, i.e., hydrothermal reaction and non-contaminating fluid sampling, were introduced subsequently. The flexible titanium foil cell was successfully tested at elevated temperature and pressure of up to 400 °C and 40 MPa, respectively, demonstrating that it could be widely used under supercritical water conditions.

  18. A new flexible titanium foil cell for hydrothermal experiments and fluid sampling

    NASA Astrophysics Data System (ADS)

    Wu, Shi-Jun; Cai, Min-Jian; Yang, Can-Jun; Li, Ke-Wei

    2016-09-01

    This paper describes the design of a flexible titanium foil cell, as well as its applications in hydrothermal experiments and in non-contaminating storage of seafloor hydrothermal fluids. A flexible cell constructed totally from pure titanium (Grade 1) can be used in corrosive environment because of the excellent chemical stability and temperature tolerance of the material. Theoretical calculation and finite element analysis of the titanium foil cell have been conducted to identify its flexibility and deformation mode. Two applications, i.e., hydrothermal reaction and non-contaminating fluid sampling, were introduced subsequently. The flexible titanium foil cell was successfully tested at elevated temperature and pressure of up to 400 °C and 40 MPa, respectively, demonstrating that it could be widely used under supercritical water conditions.

  19. Fluid shear stress activates YAP1 to promote cancer cell motility

    NASA Astrophysics Data System (ADS)

    Lee, Hyun Jung; Diaz, Miguel F.; Price, Katherine M.; Ozuna, Joyce A.; Zhang, Songlin; Sevick-Muraca, Eva M.; Hagan, John P.; Wenzel, Pamela L.

    2017-01-01

    Mechanical stress is pervasive in egress routes of malignancy, yet the intrinsic effects of force on tumour cells remain poorly understood. Here, we demonstrate that frictional force characteristic of flow in the lymphatics stimulates YAP1 to drive cancer cell migration; whereas intensities of fluid wall shear stress (WSS) typical of venous or arterial flow inhibit taxis. YAP1, but not TAZ, is strictly required for WSS-enhanced cell movement, as blockade of YAP1, TEAD1-4 or the YAP1-TEAD interaction reduces cellular velocity to levels observed without flow. Silencing of TEAD phenocopies loss of YAP1, implicating transcriptional transactivation function in mediating force-enhanced cell migration. WSS dictates expression of a network of YAP1 effectors with executive roles in invasion, chemotaxis and adhesion downstream of the ROCK-LIMK-cofilin signalling axis. Altogether, these data implicate YAP1 as a fluid mechanosensor that functions to regulate genes that promote metastasis.

  20. Fluid shear stress activates YAP1 to promote cancer cell motility

    PubMed Central

    Lee, Hyun Jung; Diaz, Miguel F.; Price, Katherine M.; Ozuna, Joyce A.; Zhang, Songlin; Sevick-Muraca, Eva M.; Hagan, John P.; Wenzel, Pamela L.

    2017-01-01

    Mechanical stress is pervasive in egress routes of malignancy, yet the intrinsic effects of force on tumour cells remain poorly understood. Here, we demonstrate that frictional force characteristic of flow in the lymphatics stimulates YAP1 to drive cancer cell migration; whereas intensities of fluid wall shear stress (WSS) typical of venous or arterial flow inhibit taxis. YAP1, but not TAZ, is strictly required for WSS-enhanced cell movement, as blockade of YAP1, TEAD1-4 or the YAP1–TEAD interaction reduces cellular velocity to levels observed without flow. Silencing of TEAD phenocopies loss of YAP1, implicating transcriptional transactivation function in mediating force-enhanced cell migration. WSS dictates expression of a network of YAP1 effectors with executive roles in invasion, chemotaxis and adhesion downstream of the ROCK–LIMK–cofilin signalling axis. Altogether, these data implicate YAP1 as a fluid mechanosensor that functions to regulate genes that promote metastasis. PMID:28098159

  1. Acute synovial fluid eosinophilia associated with delayed pressure urticaria: a role for mast cells?

    PubMed

    Miossec, P; Sullivan, T J; Tharp, M D; Volant, A; Le Goff, P

    1987-04-01

    We report a case of exercise induced joint effusion with synovial fluid (SF) eosinophilia of 9,540/mm3 in a patient with delayed pressure urticaria. The SF eosinophilia was an acute but transient event associated with some evidence of local complement activation. Histologic assessment revealed a normal synovial membrane but with no detectable intact mast cells. These observations suggest that mast cells and eosinophils acting in concert can cause joint inflammation.

  2. Increased Hepatitis E Virus Seroprevalence Correlates with Lower CD4+ Cell Counts in HIV-Infected Persons in Argentina

    PubMed Central

    Debes, José D.; Martínez Wassaf, Maribel; Pisano, María Belén; Isa, María Beatriz; Lotto, Martin; Marianelli, Leonardo G.; Frassone, Natalia; Ballari, Estefania; Bohjanen, Paul R.; Hansen, Bettina E.; Ré, Viviana

    2016-01-01

    Hepatitis E virus (HEV) is a single-stranded RNA virus that can cause hepatitis in an epidemic fashion. HEV usually causes asymptomatic or limited acute infections in immunocompetent individuals, whereas in immunosuppressed individuals such as transplant recipients, HEV can cause chronic infections. The risks and outcomes of HEV co-infection in patients infected with human immunodeficiency virus (HIV) are poorly characterized. We used a third generation immunoassay to measure serum IgG antibodies specific for HEV in 204 HIV-infected individuals from Argentina and a control group of 433 HIV-negative individuals. We found 15 of 204 (7.3%, 95%CI 3.74–10.96%) individuals in the HIV-positive group to have positive HEV IgG levels suggestive of previous infection, compared to 19 of 433 (4.4%, 95% CI 2.5–6.3%) individuals in the HIV-negative control group (p = 0.12). Among HIV-positive individuals, those with HEV seropositivity had lower CD4 counts compared to those that were HEV seronegative (average CD4 count of 234 vs 422 mm3, p = 0.01), indicating that patients with lower CD4 counts were more likely to be HEV IgG positive. Moreover, HEV seropositivity in patients with CD4 counts <200 mm3 was 16%, compared to 4.5% in those with CD4 counts >200 mm3 (p = 0.012). We found a positive PCR result for HEV in one individual. Our study found that increased seroprevalence of HEV IgG correlated with lower CD4 counts in HIV-infected patients in Argentina. PMID:27467394

  3. Effect of supplements: Probiotics and probiotic plus honey on blood cell counts and serum IgA in patients receiving pelvic radiotherapy

    PubMed Central

    Mansouri-Tehrani, Hajar-Alsadat; Rabbani-Khorasgani, Mohammad; Hosseini, Sayyed Mohsen; Mokarian, Fariborz; Mahdavi, Hoda; Roayaei, Mahnaz

    2015-01-01

    Background: Radiotherapy is frequently used in treatment approaches of pelvic malignancies. Nevertheless, it has some known systemic effects on blood cells and the immune system that possibly results in their susceptibility to infection. Probiotics are live microbial food ingredients that provide a health advantage to the consumer. Honey has prebiotic properties. The aim of this clinical trial was to investigate probable effects of probiotic or probiotics plus honey on blood cell counts and serum IgA levels in patients receiving pelvic radiotherapy. Materials and Methods: Sixty-seven adult patients with pelvic cancer were enrolled. Patients were randomized to receive either: (1) Probiotic capsules (including: Lactobacillus casei, Lactobacillus acidophilus, Lactobacillus rhamnosus, Lactobacillus bulgaricus, Bifidobacterium breve, Bifidobacterium longum, and Streptococcus thermophiles) (n = 22), (2) probiotic capsules plus honey (n = 21) or (3) placebo capsules (n = 24) all for 6 weeks. Blood and serum samples were collected for one week before radiotherapy and 24-72 h after the end of radiotherapy. Results: White blood cells (WBC), red blood cells (RBC), platelet counts, and serum IgA level were not significantly changed in patients taking probiotic (alone or plus honey) during pelvic radiotherapy. The mean decrease in RBC count was 0.52, 0.18, and 0.23 × 106 cells/μL, WBC count was 2.3, 1.21, and 1.34 × 103 cells/μL and platelet count was, 57.6, 53.3, and 66.35 × 103 cells/μL for the probiotic, probiotic plus honey, and placebo groups, respectively. The mean decrease of serum IgA was 22.53, 29.94, and 40.73 mg/dL for the probiotic, probiotic plus honey, and placebo groups, respectively. Conclusion: The observed nonsignificant effect of probiotics may be in favor of local effects of this product in the gut rather than systemic effects, however, as a trend toward a benefit was indicated, further studies are necessary in order to extract effects of probiotics or

  4. The use of computational fluid dynamic models for the optimization of cell seeding processes.

    PubMed

    Adebiyi, Adebayo A; Taslim, Mohammad E; Crawford, Keith D

    2011-12-01

    The seeding of a porous scaffold with stem cells is a fundamental step in engineering sizeable tissue constructs that are clinically viable. However, a key problem often encountered is inhomogeneous seeding of the cells particularly when the cells are delivered through the thickness of the scaffold. The objective of this study was to establish the quantitative relationships between the cell seeding efficiency and the initial vacuum pressure in a compact perfusion seeding device that uses the effect of differential pressure induced by vacuum to seed cells on a porous scaffold. A transient CFD solution of the fluid flow in the device was used to optimize the initial vacuum pressure for efficient cell seeding. Results indicate that the optimal initial vacuum pressure for homogenous cell seeding is approximately -20 kPa for the seeding device. This study presents a 3-D computational model that can be employed in designing and optimizing cell seeding techniques and corresponding technology.

  5. Modelling the damage potential of fluid flows for animal cells undergoing cultivation in bioreactors.

    NASA Astrophysics Data System (ADS)

    Stanford Keen, Giles

    1996-11-01

    Mechanical disruption and injury sustained by animal cells undergoing cultivation in bioreactors is an important problem in biotechnology. Damage to cells is thought to be caused primarily by bubbles bursting at the free surface of the culture medium. Here we present computational studies applying a mathematical model for the cell damage rates experienced by cells in laminar flow. Two fluid dynamical systems are considered - namely a converging channel and a single bursting bubble. The flows are calculated using a fourth-order finite difference technique on a stretched grid, and a boundary integral method respectively. It is possible to obtain an estimate for the number of cells in a particular population which are likely to be disrupted by the forces they experience in the flow. This is done by calculating the maximum rate of strain experienced by fluid particles, and combining this with experimental data on the strength and size of cells, obtained by micromanipulation techniques. The resulting information is then used together with the cell damage model to produce a cell damage prediction. The computational results are compared with experimental measurements of cell death, to validate the model for cell damage.

  6. Differences Between Men Who Have Sex with Men (MSM) with Low CD4 Cell Counts at Their First HIV Test and MSM with Higher CD4 Counts in Bangkok, Thailand.

    PubMed

    Sapsirisavat, Vorapot; Phanuphak, Nittaya; Sophonphan, Jiratchaya; Egan, James E; Langevattana, Kamonthip; Avihingsanon, Anchalee; Friedman, M Reuel; Stall, Ron

    2016-12-01

    Although HIV prevalence remains high among Bangkok's MSM early HIV testing as an entry point to ART has not been successfully implemented among in this population. Men who present late for initial HIV testing are a particular concern in the context of the Bangkok HIV epidemic, in that if long-term positives have had condomless sex during the time that they remained untreated they are likely to have been efficient transmitters of infection, to say nothing of the implications for their own health. A sequential sample of MSM who tested HIV positive, and CD4 counts, was taken at the Thai Red Cross Anonymous Clinic and two drop-in centers in Bangkok. Inclusion criteria were MSM aged >18 years, having not tested HIV positive earlier, who reported ≥1 of the following in the previous 6 months: condomless sex with a male, being a sex worker, or having a sexual transmitted infection (STI) diagnosis. Analysis was conducted by distinguishing between three groups of CD4 counts: <200, 200-500, >500 cells/μ to identify the social and behavioral characteristics of the men who presented late for HIV testing. Median CD4 was 325 cells/μ(n = 95). MSM with initial CD4< 200 cells/μ were significantly more likely to report problematic alcohol use. They were also more likely to report receptive anal sex and more likely to be engaged in sex work. MSM with CD4< 200 cells/μ were less likely to report recent HIV testing. Main barriers to HIV testing included being afraid of finding out that they were HIV positive and concerns about efficacy and side effects of HIV treatment. HIV stigma and concerns about treatment are still widespread and are potential barriers to HIV care among MSM in Bangkok. These barriers may work to keep men from finding out their positive HIV status in a timely manner. Thai MSM need to be made aware of the current availability of friendly HIV testing and ART services, and public health programs need to work to change their perceptions regarding ART

  7. Factors associated with the level of CD4 cell counts at HIV diagnosis in a French cohort: a quantile regression approach.

    PubMed

    Bruneau, Léa; Billaud, Eric; Raffi, François; Hanf, Matthieu

    2017-03-01

    The consensus definition of late presentation for human immunodeficiency virus patient based on a CD4 threshold of 350 cells/mm(3) has limitations concerning risk factors identification since there is growing biomedical justification for earlier initiation of treatment. The objective was to overcome this problem by simultaneously determining factors associated with different levels of CD4 counts at the time of diagnosis. Between January 2000 and July 2014, 1179 patients with a first human immunodeficiency virus diagnosis and entering care in a French human immunodeficiency virus reference center were enrolled. Factors associated with each 5 percentile from 5th to 95th quantile of CD4 counts at diagnosis were simultaneously studied in a multivariable quantile regression model. At each of the quantiles, the factors identified as negatively associated with CD4 count at diagnosis were older age, male sex , foreign patients, hepatitis B virus or hepatitis C virus co-infection, employment status, non-MSM transmission, heterosexual transmission, suburban and rural's place of residence and earlier period of diagnosis. Association with CD4 count was not uniformly significant, most factors being significant for some quantiles. The only significant determinant for all quantiles was being born in a foreign country. These results are particularly helpful in the context of human immunodeficiency virus clinical care, management and prevention.

  8. Keto analogues and amino acids supplementation induces a decrease of white blood cell counts and a reduction of muscle damage during intense exercise under thermoneutral conditions.

    PubMed

    Lima, R C P; Camerino, S R A S; França, T C L; Rodrigues, D S A; Gouveia, M G S; Ximenes-da-Silva, A; Bassini, A; Prado, E S; Cameron, L C

    2017-04-05

    This study evaluated the acute effect of keto analogue and amino acid (AA-KAAA) supplementation on both white blood cell counts and the established biomarkers of muscle damage during exercise under thermoneutral conditions. Sixteen male cyclists received a ketogenic diet for two days and were divided into two equal groups: a group taking AA-KAAA (KA) or a control group (PL). The athletes performed a two hour cycling session followed by a maximum incremental test until voluntary exhaustion (VExh). Blood samples were obtained at rest and during exercise for further hematological and biochemical analyses. Exercise-induced ammonemia increased in the PL group at VExh (75%) but remained unchanged in the KA group. Both groups exhibited a significant increase in leukocyte and neutrophil counts of ∼85% (∼13 × 10(9) L(-1)), but the shape of the lymphocytes and the eosinophil counts suggest that AA-KAAA supplementation helps prevent lymphocytosis. AA-KAAA supplementation induced a decrease in creatine kinase and aspartate aminotransferase levels at VExh while showing a significant decrease in lactate dehydrogenase at 120 min. We found that AA-KAAA supplementation decreases both the lymphocyte count response in blood and the established biomarkers of muscle damage after intense exercise under a low heat stress environment.

  9. Effect of Acute Stress on Immune Cell Counts and the Expression of Tight Junction Proteins in the Duodenal Mucosa of Rats

    PubMed Central

    Lee, Hong Sub; Kim, Dong-Kyu; Kim, Young Bae

    2013-01-01

    Background/Aims Duodenal immune alterations have been reported in a subset of patients with functional dyspepsia (FD). The aim of this study was to investigate the effect of acute stress on immune cell counts and the expression of tight junction proteins in the duodenal mucosa. Methods Twenty-one male rats were divided into the following three experimental groups: 1) the nonstressed, control group, 2) the 2-hour-stressed group, and 3) the 4-hour-stressed group. Eosinophils, mast cells and CD4+ and CD8+ T lymphocytes in the duodenal mucosa were counted. The protein and mRNA expressions of occludin and zonula occludens-1 (ZO-1) were examined. Results Eosinophils, mast cells and CD8+ T lymphocyte counts did not differ between the stressed and control groups. The number of CD4+ T lymphocytes and the protein and mRNA expressions of occludin and ZO-1 were significantly lower in the 4-hour-stressed group compared with the control group. The plasma adrenocorticotrophic hormone and cortisol levels of the 4-hour-stressed group were significantly higher than those of the control group. Conclusions Acute stress reduces the number of CD4+ T lymphocytes and the expression of tight junction proteins in the duodenal mucosa, which might be associated with the duodenal immune alterations found in a subset of FD patients. PMID:23560155

  10. Effect of dexamethasone in feed on intestinal permeability, differential white blood cell counts, and immune organs in broiler chicks.</