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1

Automated counting of white blood cells in synovial fluid  

Microsoft Academic Search

Results. The WBC count of the DIFF channel was highly correlated with the WBC count of the microscopic reference method (r ¼ 0.99; WBC analyser ¼ 0.870 ? WBC reference method þ 0.413). In contrast, no agreement existed between WBC counts generated by the WBC\\/BASO channel of the analyser and the reference method (r ¼ 0.52; WBC analyser ¼ 0.008

R. de Jonge; R. Brouwer; M. Smit; R. J. E. M. Dolhain; J. M. W. Hazes; A. W. van Toorenenbergen; J. Lindemans

2003-01-01

2

Automated white blood cell counts in cerebrospinal fluid using the body fluid mode on the platform Sysmex XE-5000.  

PubMed

Abstract Background. The Sysmex XE-5000 offers automated quantification of red blood cells and white blood cells (WBCs) in body fluids, with differentiation of polymorphonuclear cells (PMNs) and mononuclear cells (MNCs). Methods. We evaluated automated WBC counting in cerebrospinal fluid (CSF) using the body fluid mode on the Sysmex XE-5000, comparing it with flow cytometry as the reference method, and also with manual counting by microscopy. Experimental analysis for linearity and limit of detection was performed by diluting isolated WBCs in cell-free CSF. To study the ability to discriminate between PMNs and MNCs, samples were spiked using MNCs separated from peripheral blood. Comparison of WBC counts between a counting chamber and the XE-5000 was performed for 198 CSF samples. Results. In the experimental set-up, within-run (CV 19%) and between-day imprecision (CV 15.3%) in quantitating total number of WBC on XE-5000 was acceptable for WBC counts ? 25 × 10(6)/L. Compared with expected cell counts, mean bias was + 2.6% for flow cytometry, + 5.5% for XE-5000 and - 73.2% for manual counting. Differentiation between PMNs and MNCs was in concordance with flow cytometry. In comparisons of clinical CSF samples, overall agreement between the XE-5000 and manual counting was observed in 81% of the samples, but mean difference in WBC differentiation was higher for PMN (51.1 × 10(6)/L) than for MNC (7.95 × 10(6)/L). Conclusion. Despite limited precision at low WBC counts, XE-5000 could be a favourable alternative to the labour-intensive, time-consuming and less reliable manual counting and cuts turnaround times in routine CSF-based diagnosis. PMID:25180445

Li, Aihong; Grönlund, Elisabeth; Brattsand, Göran

2014-11-01

3

Evaluation of White Cell Count and Differential in Synovial Fluid for Diagnosing Infections after Total Hip or Knee Arthroplasty  

PubMed Central

Background The accuracy of synovial fluid (SF) white cell count (WCC) and polymorphonuclear (PMN) cell evaluation for predicting prosthetic joint infection (PJI) at the total hip arthroplasty (THA) or total knee arthroplasty (TKA) site is unknown. Therefore, we performed a meta-analysis to summarize the diagnostic validity of SF-WCC and SF-PMN for diagnosing PJI. Methods The MEDLINE, EMBASE, and OVID databases were searched for studies that had evaluated the diagnostic validity of SF-WCC and SF-PMN between January 1990 and May 2013. Meta-analysis methods were used to pool sensitivity, specificity, diagnostic odd ratios (DORs), the area under the receiver-operating characteristic curve (AUC), positive likelihood ratios (PLR), negative likelihood ratios (NLR), and post-test probability. We also conducted heterogeneity, publication bias, subgroup, and meta-regression analyses. Results Fifteen articles (15 SF-WCC and 14 SF-PMN) that included a total of 2787 patients fulfilled the inclusion criteria and were considered for analysis. The pooled sensitivity and specificity for PJI detection was 0.88 (95% confidence intervals [CI], 0.81–0.93) and 0.93 (95% CI, 0.88–0.96) for SF-WCC and 0.90 (95% CI, 0.84–0.93) and 0.88 (95% CI, 0.83–0.92) for SF-PMN, respectively. The AUC was 0.96 for SF-WCC and 0.95 for SF-PMN. PLR and NLR were 13.3 and 0.13 for SF-WCC, and 7.6 and 0.12 for SF-PMN, respectively. There was no evidence of publication bias. Low-clinical-scenario (pre-test probability, 20%) post-test probabilities were 3% for both negative SF-WCC and SF-PMN results. The subgroup analyses indicated that the sensitivity/specificity of THA were 0.73/0.96 for SF-WCC and 0.85/0.83 for SF-PMN, whereas those of TKA were 0.90/0.91 for SF-WCC and 0.90/0.88 for SF-PMN. We also found that collection of SF-WCC preoperatively had a higher sensitivity than that obtained intraoperatively (0.91 vs. 0.77). Conclusions SF-WCC and SF-PMN have an adequate and clinically acceptable diagnostic value for detecting PJI, particularly after TKA. PMID:24416276

Li, Haowei; Wu, Chuanlong; Li, Yang; Li, Huiwu; Zhu, Zhenan; Qin, An; Dai, Kerong

2014-01-01

4

White blood cell count - series (image)  

MedlinePLUS

The White Blood Cell (WBC) Count measures two components: the total number of WBC's (leukocytes), and the differential count. ... and basophils) and non-granulocytes (lymphocytes and monocytes). White blood cells are a major component of the ...

5

Somatic cell counts of ewes' milk.  

PubMed

The somatic cell counts of ewes' milk were determined by an electronic particle counter (Coulter Counter). Of 1408 apparently normal milk samples, 98.2% had a somatic cell count lower than 1.0 x 10(6) cells/ml and 85.8% of 254 bacteriologically positive samples had a count higher than 1.0 x 10(6) cells/ml. Values exceeding 1.0 x 10(6) cells/ml are indicative of subclinical mastitis, if samples were collected from clinically healthy mammary glands. PMID:1777802

Fthenakis, G C; el-Masannat, E T; Booth, J M; Jones, J E

1991-01-01

6

21 CFR 864.6160 - Manual blood cell counting device.  

Code of Federal Regulations, 2010 CFR

... 2010-04-01 false Manual blood cell counting device. 864.6160 Section...Hematology Devices § 864.6160 Manual blood cell counting device. (a) Identification. A manual blood cell counting device is a device used to...

2010-04-01

7

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

E-print Network

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

8

Why Count Types of White Blood Cells?  

NSDL National Science Digital Library

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

Ethel D. Stanley (Beloit College;Biology); Donald Buckley (Quinnipiac University;Biology)

2006-05-20

9

Differential white cell count by centrifugal microfluidics.  

SciTech Connect

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.

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

2010-07-01

10

Counting  

NSDL National Science Digital Library

Practicing Counting Count the number of bunnies and match it to the correct number of characters on the right. Click on the correct one. Bunny Count Practice counting by 1 Countin by 1 Practice counting Sea Horses! Counting Sea Horses ...

Person, Ms.

2007-11-28

11

Rapid count of microbial cells in dialysate.  

PubMed

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

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

2007-10-01

12

Electrical cell counting process characterization in a microfluidic impedance cytometer  

E-print Network

measurement channel used to electrically count the cells. We show that cell counting over time is a non-homogeneous in milk (Phipps and Newbould 1996). Other examples include, particle size determination in composition

Bashir, Rashid

13

Comparison between a modified haemocytometric technique and electronic counters in goat blood cell counting.  

PubMed

Dilutions of goat blood with Hayem-Jørgensen's fluid ranging from 1:200 to 1:1,000 were used for haemocytometer counting of red blood cells (RBC) in 27 goats. The optimal dilutions were 1:400-1:500. Correlation studies between the results obtained by the haemocytometer and the Coulter counter red blood cell (RBC) and white blood cell (WBC) counts were performed in 551 goat blood samples. The haemocytometer RBC counts were 5.63% higher and WBC counts 2.79% lower than those of the electronic counter. The method of blood cell counting therefore influences the clinical haematological diagnoses and reference values in domestic animals. New cell counters specifically designed to measure cells of small volumes, e.g. goat erythrocytes, are needed. PMID:1910237

Mbassa, G K; Poulsen, J S

1991-06-01

14

Counting  

NSDL National Science Digital Library

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

Beck, Mrs.

2006-12-08

15

Photon Counts Statistics in Leukocyte Cell Dynamics  

NASA Astrophysics Data System (ADS)

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.

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

2011-12-01

16

Cell Counts in Cerebral Cortex of an Autistic Patient.  

ERIC Educational Resources Information Center

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

Coleman, Paul D.; And Others

1985-01-01

17

RAPID GLUTARALDEHYDE FIXATION FOR COW MILK CELL COUNT  

E-print Network

of milk ; a ratio of 2.5 : 1000 (V/V). Samples were homogenized and then kept at laboratory temperatureRAPID GLUTARALDEHYDE FIXATION FOR COW MILK CELL COUNT BY COULTER COUNTER B. POUTREL G. DUBRAY INRA) is regarded as the reference method for counting somatic cells in cow milk. The use of an electronic particle

Paris-Sud XI, Université de

18

Cell counts in cerebral cortex of an autistic patient  

Microsoft Academic Search

Numbers of neurons and glia were counted in the cerebral cortex of one well-documented case of autism and two age and sexmatched controls. Areas in which cell counts were made were primary auditory cortex, Broca's speech area, and auditory association cortex. No consistent differences in cell density were found between the brains of the autistic patient and the control patients.

Paul D. Coleman; John Romano; Lowell Lapham; William Simon

1985-01-01

19

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

PubMed Central

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

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

20

21 CFR 864.6160 - Manual blood cell counting device.  

... FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES HEMATOLOGY AND PATHOLOGY DEVICES Manual Hematology Devices § 864.6160 Manual blood cell counting device. (a) Identification. A...

2014-04-01

21

21 CFR 864.6160 - Manual blood cell counting device.  

Code of Federal Regulations, 2011 CFR

... FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES HEMATOLOGY AND PATHOLOGY DEVICES Manual Hematology Devices § 864.6160 Manual blood cell counting device. (a) Identification. A...

2011-04-01

22

21 CFR 864.6160 - Manual blood cell counting device.  

Code of Federal Regulations, 2012 CFR

... FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES HEMATOLOGY AND PATHOLOGY DEVICES Manual Hematology Devices § 864.6160 Manual blood cell counting device. (a) Identification. A...

2012-04-01

23

21 CFR 864.6160 - Manual blood cell counting device.  

Code of Federal Regulations, 2013 CFR

... FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES HEMATOLOGY AND PATHOLOGY DEVICES Manual Hematology Devices § 864.6160 Manual blood cell counting device. (a) Identification. A...

2013-04-01

24

A system for counting fetal and maternal red blood cells.  

PubMed

The Kleihauer-Betke (KB) test is the standard method for quantitating fetal-maternal hemorrhage in maternal care. In hospitals, the KB test is performed by a certified technologist to count a minimum of 2000 fetal and maternal red blood cells (RBCs) on a blood smear. Manual counting suffers from inherent inconsistency and unreliability. This paper describes a system for automated counting and distinguishing fetal and maternal RBCs on clinical KB slides. A custom-adapted hardware platform is used for KB slide scanning and image capturing. Spatial-color pixel classification with spectral clustering is proposed to separate overlapping cells. Optimal clustering number and total cell number are obtained through maximizing cluster validity index. To accurately identify fetal RBCs from maternal RBCs, multiple features including cell size, roundness, gradient, and saturation difference between cell and whole slide are used in supervised learning to generate feature vectors, to tackle cell color, shape, and contrast variations across clinical KB slides. The results show that the automated system is capable of completing the counting of over 60 000 cells (versus  ? 2000 by technologists) within 5 min (versus  ? 15 min by technologists). The throughput is improved by approximately 90 times compared to manual reading by technologists. The counting results are highly accurate and correlate strongly with those from benchmarking flow cytometry measurement. PMID:24879644

Ge, Ji; Gong, Zheng; Chen, Jun; Liu, Jun; Nguyen, John; Yang, Zongyi; Wang, Chen; Sun, Yu

2014-12-01

25

Development of a stained cell nuclei counting system  

NASA Astrophysics Data System (ADS)

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.

Timilsina, Niranjan; Moffatt, Christopher; Okada, Kazunori

2011-03-01

26

Making stem cells count for global health.  

PubMed

Developing countries such as China, India and Brazil are making large investments in the stem cell field. Here we argue that hands-on involvement in the field by these countries is essential if the products developed are going to be locally relevant, affordable and appropriate. However, stem cells are a high-risk investment and any global health impacts are still likely to be far off. Even if they are eventually successful, better clinical oversight and measures to ensure access are required for stem cells to have a substantial and equitable impact. PMID:21999282

McMahon, Dominique S; Thorsteinsdóttir, Halla

2011-11-01

27

WBC (White Blood Cell) Differential Count  

MedlinePLUS

... occur with bacterial infection, leukemia , myelodysplastic disorders, or myeloproliferative neoplasms , for example. Some immature cells that may be ... g., autoimmune disorders , immune deficiency) Leukemia Myelodysplastic syndrome Myeloproliferative neoplasms Some diseases trigger a response by the immune ...

28

Pediatric metabolic syndrome and cell blood counts: bivariate Bayesian modeling.  

PubMed

Cell blood counts are components of hematological parameters and indicators of pro-inflammatory states. They are proposed to be associated with metabolic syndrome (MetS). This study aimed to assess the relationship of the white blood cell (WBC) and the red blood cell (RBC) counts with components of MetS in the pediatric age group. The sample consisted of 300 children (152 boys) aged 6-12 years. Hierarchical Bayesian analysis of the bivariate Poisson regression model was used to estimate the effect of various components of MetS according to the cell blood counts. We found that RBC and WBC counts were correlated with the fasting blood glucose, the waist-to-height ratio, serum triglycerides and the blood pressure levels adjusted for age, the body mass index, gender, total cholesterol, low-density lipoprotein cholesterol and the hip circumference. The high-density lipoprotein cholesterol was correlated with the RBC counts based on 95% high posterior density regions for parameters in the Bayesian model. Our findings may serve as confirmatory evidence for the beginning of inflammatory process related to the cardio-metabolic factors from early life. PMID:24108065

Mansourian, Marjan; Kazemi, Iraj; Kelishadi, Roya

2014-02-01

29

Dynamics and regulation of bulk milk somatic cell counts.  

PubMed Central

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

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

1993-01-01

30

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

Code of Federal Regulations, 2010 CFR

...2010-04-01 2010-04-01 false Calibrator for red cell and white cell counting. 864.8185 Section 864.8185 Food...Hematology Reagents § 864.8185 Calibrator for red cell and white cell counting. (a) Identification....

2010-04-01

31

Interaural Comparison of Spiral Ganglion Cell Counts in Profound Deafness  

PubMed Central

Objectives This study is designed to measure the degree to which spiral ganglion cell (SGC) survival in the left and right ears is similar in profoundly hearing-impaired human patients with symmetric (right/left) etiology and sensitivity. This is of interest because a small difference between ears would imply that one ear could be used as a control ear in temporal bone studies evaluating the impact on SGC survival of a medical intervention in the other ear. Materials and Methods Forty-two temporal bones from 21 individuals with bilaterally symmetric profound hearing impairment were studied. Both ears in each individual were impaired by the same etiology. Rosenthal’s canal was reconstructed in two dimensions and segmental and total SGCs were counted. Correlation analysis and t-tests were used to compare segmental and total counts of left and right ears. Statistical power calculations illustrate how the results can be used to estimate the effect size (right/left difference in SGC count) that can be reliably identified as a function of sample size. Results Left counts (segmental and total) were significantly correlated with those in the right ears (p<0.01) and the coefficients of determination for segments 1 to 4 and total count were respectively 0.64, 0.91, 0.93, 0.91 and 0.98. The hypothesis that mean segmental and total counts of right and left are the same could not be rejected by paired t-test. Conclusion The variance in the between-ear difference across the temporal bones studied indicates that useful effect sizes can be reliably identified using subject numbers that are practical for temporal bone studies. For instance, there is 95% likelihood that an interaural difference in SGC count of approximately 1000 cells associated with a treatment/manipulation of one ear will be reliably detected in a bilaterally-symmetric profound hearing loss population of temporal bones from approximately 10 subjects. PMID:22008826

Seyyedi, Mohammad; Eddington, Donald K; Nadol, Joseph B

2011-01-01

32

Prognostic Value of Elevated White Blood Cell Count in Hypertension  

Microsoft Academic Search

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

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

2007-01-01

33

Digital Cell Counting Device Integrated with a Single-Cell Array  

PubMed Central

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 mm2 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 (r2?=?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. PMID:24551208

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

2014-01-01

34

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

... FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES HEMATOLOGY AND PATHOLOGY DEVICES Hematology Reagents § 864.8185 Calibrator for red cell and white cell counting. (a)...

2014-04-01

35

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

Code of Federal Regulations, 2011 CFR

... FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES HEMATOLOGY AND PATHOLOGY DEVICES Hematology Reagents § 864.8185 Calibrator for red cell and white cell counting. (a)...

2011-04-01

36

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

Code of Federal Regulations, 2012 CFR

... FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES HEMATOLOGY AND PATHOLOGY DEVICES Hematology Reagents § 864.8185 Calibrator for red cell and white cell counting. (a)...

2012-04-01

37

Counting Legionella Cells within Single Amoeba Host Cells  

PubMed Central

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

Ashbolt, Nicholas J.

2012-01-01

38

Counting Legionella cells within single amoeba host cells.  

PubMed

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

Buse, Helen Y; Ashbolt, Nicholas J

2012-03-01

39

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...

40

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

PubMed

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. PMID:24529513

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

2014-05-01

41

WBC count  

MedlinePLUS

Leukocyte count; White blood cell count ... in the blood is 4,500-10,000 white blood cells per microliter (mcL). Normal value ranges ... LOW WHITE BLOOD CELL (WBC) COUNT A low number of WBCs is called leukopenia. A WBC count below 4500 ...

42

Differential leucocyte cell counts from the pygoscelid penguins of Antarctica.  

PubMed

Differential leucocyte counts were obtained for three cogeneric species of wild antarctic penguins, Pygoscelis adelie (adelie), Pygoscelis papua (gentoo), and Pygoscelis antarctica (chinstrap). Significant differences between the differential leucocyte counts of the three species were not observed. PMID:3625920

Zinsmeister, V A; VanDerHeyden, M J

1987-07-01

43

Transient Transfection of Suspension Cells by Electroporation 1. Count cells. The cells should be in the exponential phase of growth.  

E-print Network

Transient Transfection of Suspension Cells by Electroporation 1. Count cells. The cells should be in the exponential phase of growth. 2. Collect 4 x 106 cells per transfection. Wash cells twice with Opti-MEM media. NOTE: Omit Penicillin-Streptomycin from the Opti-MEM transfection media, as electroporated cells

Gauthier, Eric

44

Interpretation of Changes in Circulating Tumor Cell Counts12  

PubMed Central

The presence of circulating tumor cells (CTCs) in the blood of cancer patients may guide the use of therapy. We investigated how to evaluate a reduction in the number of CTCs after administration of therapy. CTCs were enumerated with the CellSearch system in 111 metastatic breast and 185 metastatic prostate cancer patients before start of a new line of chemotherapy and after initiation of therapy. Different means to express changes in CTC counts were evaluated with respect to overall survival (OS). A static CTC cutoff is the best method to determine whether a therapy is effective. This is exemplified by the highest Cox hazard ratio of 2.1 for OS; three methods to express relative differences performed worse. A lookup table is provided from which the significance of a change in CTCs can be derived. The aim of therapy should be the elimination of all CTCs. A period of 10 to 12 weeks of therapy is needed to reach the treatment effect on CTCs. PMID:23323160

Coumans, Frank AW; Ligthart, Sjoerd T; Terstappen, Leon WMM

2012-01-01

45

CD117+ amniotic fluid stem cells  

PubMed Central

Broadly multipotent stem cells can be isolated from amniotic fluid by selection for the expression of the membrane stem cell factor receptor c-Kit, a common marker for multipotential stem cells. They have clonogenic capability and can be directed into a wide range of cell types representing the three primary embryonic lineages. Amniotic fluid stem cells maintained for over 250 population doublings retained long telomeres and a normal karyotype. Clonal human lines verified by retroviral marking were induced to differentiate into cell types representing each embryonic germ layer, including cells of adipogenic, osteogenic, myogenic, endothelial, neuronal and hepatic lineages. AFS cells could be differentiate toward cardiomyogenic lineages, when co-cultured with neonatal cardiomyocytes, and have the potential to generate myogenic and hematopoietic lineages both in vitro and in vivo. Very recently first trimester AFS cells could be reprogrammed without any genetic manipulation opening new possibilities in the field of fetal/neonatal therapy and disease modeling. In this review we are aiming to summarize the knowledge on amniotic fluid stem cells and highlight the most promising results. PMID:23037870

Cananzi, Mara; De Coppi, Paolo

2012-01-01

46

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

Microsoft Academic Search

White cell counts at rest might be lower in athletes participating in selected endurance-type sports. Here, we analysed blood\\u000a tests of elite athletes collected over a 10-year period. Reference ranges were established for 14 female and 14 male sports\\u000a involving 3,679 samples from 937 females and 4,654 samples from 1,310 males. Total white blood cell counts and counts of neutrophils,

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

2010-01-01

47

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

PubMed Central

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

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

48

Enhancing the performance of a point-of-care CD4+ T-cell counting microchip through monocyte depletion for HIV/AIDS diagnostics  

PubMed Central

CD4+ T cell counts are important tests used to stage HIV-postive patients, enabling clinicians to make informed antiretroviral treatment decisions and to monitor the therapeutic outcomes. However, state-of-the-art CD4 counting methods based on flow cytometry are not applicable in resource-limited settings, due to their high cost and technical requirements. In previous work, we reported the development of a cell isolation microchip that can be used at the point of care for CD4 counts. In that microfluidic chip, CD4+ T cells were separated from 10 ?L of whole blood, and enumerated via either light microscopy or impedance sensing. The microchip counts matched flow cytometry results in the intermediate CD4 count range, between 200–800 cells/?L, but displayed a positive bias at absolute CD4 counts below 200 cells/?L, due largely to monocyte contamination. To enhance the performance in the low CD4 count range, we report here an improved design of a two-stage microfluidic device to deplete monocytes from whole blood, followed by CD4+ T cell capture. Using the double-stage device combined with a high viscosity rinsing solution, we obtained microchip CD4 counts comparable to flow cytometry results in the full clinically relevant range. In addition to CD4 counting, the strategy of contaminant depletion prior to target cell isolation can be easily adapted to immunoaffinity capture of other cell types that lack a unique surface marker from a complex biological fluid. PMID:19417901

Cheng, Xuanhong; Gupta, Amit; Chen, Chihchen; Tompkins, Ronald G.; Rodriguez, William; Toner, Mehmet

2014-01-01

49

Dynamic changes in white blood cell counts in uncomplicated Plasmodium falciparum and P. vivax malaria  

Microsoft Academic Search

Total and differential white blood cell (WBC) counts are basic and essential indicators in any type of illness resulting from infection. In malaria, WBC counts are generally characterized as low to normal during treatment. WBC-counts data, before and during treatment with artemisinin derivatives, was gathered for patients with either Plasmodium falciparum or Plasmodium vivax infection (at 28-day follow-up), to investigate

Noppadon Tangpukdee; Haur-Sen Yew; Srivicha Krudsood; Nataya Punyapradit; Waraporn Somwong; Sornchai Looareesuwan; Shigeyuki Kano; Polrat Wilairatana

2008-01-01

50

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

PubMed Central

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

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

2014-01-01

51

Optimization of a cell counting algorithm for mobile point-of-care testing platforms.  

PubMed

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

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

2014-01-01

52

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

PubMed Central

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

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

2014-01-01

53

Pseudospontaneous Platelet Aggregation and Spuriously Elevated Blood Cell Counts in Cryoglobulinemia  

Microsoft Academic Search

Cryoglobulinemia has been reported sometimes to cause false elevation in platelet and\\/or leukocyte counts in au tomated blood cell counters due to temperature-dependent pro tein precipitates that are falsely interpreted as blood cells at room temperature. Upon heating to 37°C, these blood counts decrease to normal levels as cryoproteins dissolve. Therefore, cryoglobulins could theoretically lead to erroneous diagnosis of spontaneous

Yahya Büyüka?ik; N. ?emnur ileri; Ibrahim C. Haznedaroglu; Selma Karaahmetoglu; Osman Müftüo?lu; ?erafettin Kirazli; Semra Dündar

1998-01-01

54

On Efficient Query Processing of Stream Counts on the Cell Processor  

Microsoft Academic Search

In recent years, the sketch-based technique has been presented as an effective method for counting stream items on processors with limited storage and processing capabilities, such as the network processors. In this paper, we examine the implementation of a sketch-based counting algorithm on the heterogeneous multi-core Cell processor. Like the network processors, the Cell also contains on-chip special processors with

Dina Thomas; Rajesh Bordawekar; Charu C. Aggarwal; Philip S. Yu

2009-01-01

55

The prevalence of some mastitis pathogens in bulk milk of Dutch dairy goats and the relationship with bulk milk somatic cell count and bulk milk standard plate count  

Microsoft Academic Search

The aim of this study was to assess the correlation of bulk milk somatic cell count (BMSCC) with bulk milk standard plate count (BMSPC) and with the number of coliform bacteria, coagulase negative staphylococci (CNS) and S. aureus . We also described the prevalence of these pathogens in bulk milk of Dutch dairy goat farms. In total 53 dairy goat

N. Dik; G. Koop; L. Lipman

56

Leukocyte Count in the Synovial Fluid of Children with Culture-Proven Brucellar Arthritis  

Microsoft Academic Search

:   Brucellosis is an important cause of paediatric septic arthritis in endemic areas. Because the Gram stain is frequently negative\\u000a and culture results are unavailable at the time of the patient’s admission, the diagnosis of brucellar arthritis is usually\\u000a entertained on the bases of epidemiological considerations and cytological examination of the synovial fluid aspirate. The\\u000a aim of this study was

N. Peled; D. Buskila; P. Yagupsky

2002-01-01

57

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

PubMed

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. PMID:20640439

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

2010-11-01

58

Mechanotransduction of fluid stresses governs 3D cell migration  

E-print Network

Solid tumors are characterized by high interstitial fluid pressure, which drives fluid efflux from the tumor core. Tumor-associated interstitial flow (IF) at a rate of ?3 µm/s has been shown to induce cell migration in the ...

Polacheck, William J.

59

Differences in estimates of cisplatin-induced cell kill in vitro between colorimetric and cell count/colony assays.  

PubMed

The aim of this study was to evaluate some bioassays that are different in principle: cell counting, colony forming assay, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide (MTT), sulforhodamine B (SRB), crystal violet, and alamarBlue, with respect to their ability to measure cisplatin-induced cell death of in vitro-cultivated squamous cell carcinoma of the head and neck (SCCHN). Cisplatin was applied in concentrations of 1.0, 5.0, 10.0, 50.0, and 100 microM. The cells were incubated for 1 h, and the cell survival was measured 5 d after treatment. We found the colorimetric assays and cell counting to be comparable. The colony forming assay indicated a higher degree of cell kill compared with the other techniques. Measurement of cell survival after treatment with cisplatin can be done by use of any of the above tested assays. However, the majority of SCCHN cell lines available do not form colonies easily, or at all. Therefore, comparing the chemosensitivity between such cell lines is limited to alternative assays. In this respect, any of the tested colorimetric assays can be used. However, they seem to underestimate cell kill. Cell counting is also an alternative. This technique, however, is time consuming and operator dependent, as in the case of manual counting, or relatively expensive when counting is performed electronically, compared with the colorimetric assays. PMID:17316066

Henriksson, Eva; Kjellén, Elisabeth; Wahlberg, Peter; Wennerberg, Johan; Kjellström, Johan H

2006-01-01

60

Nucleated Red Blood Cells Count in Pregnancies with Idiopathic Intra-Uterine Growth Restriction  

PubMed Central

Objective Elevated nucleated red blood cell (NRBC) count is introduced as a potential marker of intra-uterine growth restriction (IUGR). To investigate the probable association regardless of any known underlying disease, we aimed to study disturbances in NRBC count in infants experiencing idiopathic IUGR. Materials and methods Twenty three infants regarded IUGR without any known cause were chosen to be compared to 48 normal neonates. Blood samples were collected instantly after birth and the same measurements were done in both groups. Results NRBC count/100 white blood cells was significantly higher in the IUGR group (P value < 0.001). pH measurements did not reveal any significant difference. Conclusion Increased NRBC count in cases of idiopathic IUGR in absence of chronic hypoxia could strengthen its predictive value suggested in previous studies. It could help early IUGR detection and beneficial intervention. PMID:24971139

Kaveh, Mahbod; Nemati, Somayeh; Javadian, Pouya; Salmanian, Bahram

2014-01-01

61

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

PubMed

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

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

2014-12-01

62

MICRO/MACROSCOPIC FLUID FLOW IN OPEN CELL FIBROUS STRUCTURES  

E-print Network

MICRO/MACROSCOPIC FLUID FLOW IN OPEN CELL FIBROUS STRUCTURES by Ali Tamayol M.Sc., Sharif: Doctor of Philosophy (PhD) Title of Thesis: MICRO/MACROSCOPIC FLUID FLOW IN OPEN CELL FIBROUS STRUCTURES including composite fabrication, filtration, compact heat exchangers, fuel cell technology, and tissue

Bahrami, Majid

63

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

PubMed Central

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?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 cells, with particular relevance to translation in the clinic, where a Phase I clinical trial of DARC in glaucoma patients is due to start shortly. PMID:24902592

2014-01-01

64

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

PubMed Central

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

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

2013-01-01

65

Antimicrobial susceptibility testing in 90 min by bacterial cell count monitoring  

PubMed Central

The rise in antimicrobial resistance has become a serious global health problem. Restrictive use of antibiotics seems the only option to temper this accession since research in new antibiotics has halted. Antimicrobial stewardship programmes rely on quick access to susceptibility data. This study evaluated the concept of bacterial cell count monitoring as a fast method to determine susceptibility. Escherichia coli, Pseudomonas aeruginosa and Staphylococcus aureus strains were tested for amoxicillin/piperacillin and gentamicin by three conventional methods (VITEK2®, Etest® and broth-macrodilution). Bacterial cell count monitoring reliably predicted susceptibility after 90 min for Escherichia coli and after 120 min for Pseudomonas aeruginosa and Staphylococcus aureus without any minor, major or very major discrepancies. Time-to-result was reduced by 74%, 83% and 76%, respectively. Bacterial cell count monitoring shows great potential for rapid susceptibility testing. PMID:22390723

Broeren, M A C; Maas, Y; Retera, E; Arents, N L A

2013-01-01

66

Complete blood cell count in psittaciformes by using high-throughput image cytometry: a pilot study.  

PubMed

The avian hemogram is usually performed in veterinary diagnostic laboratories by using manual cell counting techniques and differential counts determined by light microscopy. There is no standard automated technique for avian blood cell count and differentiation to date. These shortcomings in birds are primarily because erythrocytes and thrombocytes are nucleated, which precludes the use of automated analyzers programmed to perform mammal complete blood cell counts. In addition, there is no standard avian antibody panel, which would allow cell differentiation by immunophenotyping across all commonly seen bird species. We report an alternative hematologic approach for quantification and differentiation of avian blood cells by using high-throughput image cytometry on blood smears in psittacine bird species. A pilot study was designed with 70 blood smears of different psittacine bird species stained with a Wright-Giemsa stain. The slides were scanned at 0.23 microm/pixel. The open-source softwares CellProfiler and CellProfiler Analyst were used for analyzing and sorting each cell by image cytometry. A "pipeline" was constructed in the CellProfiler by using different modules to identify and export hundreds of measures per cell for shape, intensity, and texture. Rules for classifying the different blood cell phenotypes were then determined based on these measurements by iterative feedback and machine learning by using CellProfiler Analyst. Although this approach shows promises, avian Leukopet results could not be duplicated when using this technique as is. Further studies and more standardized prospective investigations may be needed to refine the "pipeline" strategy and the machine learning algorithm. PMID:24344512

Beaufrère, Hugues; Ammersbach, Mélanie; Tully, Thomas N

2013-09-01

67

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

PubMed Central

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

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

2014-01-01

68

Electrical Counting and Sizing of Mammalian Cells in Suspension  

PubMed Central

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

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

1965-01-01

69

Finite Element Analyses of Fluid Flow Conditions in Cell Culture  

PubMed Central

Numerous studies in tissue engineering and biomechanics use fluid flow stimulation, both unidirectional and oscillatory, to analyze the effects of shear stresses on cell behavior. However, it has typically been assumed that these shear stresses are uniform and that cell and substrate properties do not adversely affect these assumptions. With the increasing utilization of fluid flow in cell biology, it would be beneficial to determine the validity of various experimental protocols. Because it is difficult to determine the velocity profiles and shear stresses empirically, we used the finite element method (FEM). Using FEM, we determined the effects of cell confluence on fluid flow, the effects of cell height on the uniformity of shear stresses, apparent shear stresses exhibited by cells cultured on various substrates, and the effects of oscillatory fluid flow relative to the unidirectional flow. FEM analyses could successfully analyze flow patterns over cells for various cell confluence and shape and substrate characteristics. Our data suggest the benefits of the utilization of oscillatory fluid flow and the use of substrates that stimulate cell spreading in the distribution of more uniform shear stresses across the surface of cells. Also we demonstrated that the cells cultured on nanotopographies were exposed to greater apparent shear stresses than cells on flat controls when using the same fluid flow conditions. FEM thus provides an excellent tool for the development of experimental protocols and the design of bioreactor systems. PMID:19778171

Salvi, Joshua D.; Lim, Jung Yul

2010-01-01

70

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

PubMed Central

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

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

2014-01-01

71

Which observations from the complete blood cell count predict mortality for hospitalized patients?  

PubMed Central

Background Information on the prognostic utility of the admission complete blood count (CBC) and differential count is lacking. Objective To identify independent predictors of mortality from the varied number and morphology of cells in the complete blood count defined as a hemogram, automated five cell differential count and manual differential count. Design Retrospective cohort study and chart review. Setting Wishard Memorial Hospital, a large urban primary care hospital. Patients 46,522 adult inpatients admitted over ten years to Wishard Memorial Hospital from January 1993 through December of 2002. Intervention None Measurements 30-day mortality measured from day of admission as determined by electronic medical records and Indiana State Death records. Results Controlling for age and gender, the multivariable regression model identified three strong independent predictors of 30-day mortality: Presence of nucleated RBCs, burr cells, or absolute lymphocytosis was associated with a three-fold increased risk of mortality at 30 days. Nucleated RBCs were associated with a 25.5% 30-day mortality rate across a range of diagnoses, excluding sickle cell disease and obstetrical patients in which NRBCs were not associated with increased mortality. Burr cells were associated with a 27.3% mortality rate and found most commonly in patients with renal failure or liver failure. Absolute lymphocytosis predicted a poor outcome in trauma and CNS injury patients. Conclusions In patients admitted to the hospital, presence of nucleated RBCs, burr cells, or absolute lymphocytosis at admission is each associated with a three-fold increase in risk of 30-day mortality. PMID:17274042

Kho, Abel N; Hui, Siu; Kesterson, Joe G.; McDonald, Clement J

2013-01-01

72

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

E-print Network

Computer-assisted counting of retinal cells by automatic segmentation after TV denoising Kristian Bredies, Marcus Wagner, Christian Schubert and Peter Ahnelt Abstract. Background. Quantitative evaluation on Rudin-Osher-Fatemi total variation (TV) denoising. Once 3 parameters are manually adjusted based

Wagner, Marcus

73

Increased mast cell counts in benign and malignant salivary gland tumors.  

PubMed

Background and aims. Mast cells are one of the characteristic factors in angiogenesis, growth, and metastatic spread of tumors. The distribution and significance of mast cells in many tumors have been demonstrated. However, few studies have evaluated mast cell infiltration in salivary gland tumors. In this study, mast cell counts were evaluated in benign and malig-nant salivary gland tumors. Materials and methods. This descriptive and cross-sectional study assessed 30 cases of pleomorphic adenoma, 13 cases of adenoid cystic carcinoma, 7 cases of mucoepidermoid carcinoma (diagnosed on the basis of 2005 WHO classifica-tion), with adequate stroma in peritumoral and intratumoral areas, and 10 cases of normal salivary glands. The samples were stained with 5% diluted Giemsa solution and the average stained cell counts were calculated in 10 random microscopic fields in peri- and intra-tumoral areas. Data were analyzed by t-test and Mann-Whitney and Krusskal-Wallis tests. Results. The average mast cell counts increased in the tumors compared to normal salivary glands. There was no signifi-cant difference between benign and malignant tumors and also between different malignant tumors. Infiltration was signifi-cantly denser in peri-tumoral stroma in both tumoral groups (P = 0.001). Minor salivary glands contained significantly more numerous mast cells. Conclusion. Although mast cell counts increased in benign and malignant salivary gland tumors, there were no signifi-cant differences between the tumoral groups. Further studies are suggested to determine the type of these cells which might be useful in the assessment of biological nature of the tumor and its future treatment modality. PMID:25024834

Jaafari-Ashkavandi, Zohreh; Ashraf, Mohammad-Javad

2014-01-01

74

Increased Mast Cell Counts in Benign and Malignant Salivary Gland Tumors  

PubMed Central

Background and aims. Mast cells are one of the characteristic factors in angiogenesis, growth, and metastatic spread of tumors. The distribution and significance of mast cells in many tumors have been demonstrated. However, few studies have evaluated mast cell infiltration in salivary gland tumors. In this study, mast cell counts were evaluated in benign and malig-nant salivary gland tumors. Materials and methods. This descriptive and cross-sectional study assessed 30 cases of pleomorphic adenoma, 13 cases of adenoid cystic carcinoma, 7 cases of mucoepidermoid carcinoma (diagnosed on the basis of 2005 WHO classifica-tion), with adequate stroma in peritumoral and intratumoral areas, and 10 cases of normal salivary glands. The samples were stained with 5% diluted Giemsa solution and the average stained cell counts were calculated in 10 random microscopic fields in peri- and intra-tumoral areas. Data were analyzed by t-test and Mann-Whitney and Krusskal-Wallis tests. Results. The average mast cell counts increased in the tumors compared to normal salivary glands. There was no signifi-cant difference between benign and malignant tumors and also between different malignant tumors. Infiltration was signifi-cantly denser in peri-tumoral stroma in both tumoral groups (P = 0.001). Minor salivary glands contained significantly more numerous mast cells. Conclusion. Although mast cell counts increased in benign and malignant salivary gland tumors, there were no signifi-cant differences between the tumoral groups. Further studies are suggested to determine the type of these cells which might be useful in the assessment of biological nature of the tumor and its future treatment modality. PMID:25024834

Jaafari-Ashkavandi, Zohreh; Ashraf, Mohammad-Javad

2014-01-01

75

FISH and chips: automation of fluorescent dot counting in interphase cell nuclei.  

PubMed

Fluorescence in situ hybridization allows the enumeration of chromosomal abnormalities in interphase cell nuclei. This process is called dot counting. To estimate the distribution of chromosomes per cell, a large number of cells have to be analyzed, especially when the frequency of aberrant cells is low. Automation of dot counting is required because manual counting is tedious, fatiguing, and time-consuming. We developed a completely automated fluorescence microscope system that can examine 500 cells in approximately 15 min to determine the number of labeled chromosomes (seen as dots) in each cell nucleus. This system works with two fluorescent dyes, one for the DNA hybridization dots and one for the cell nucleus. After the stage has moved to a new field, the image is automatically focused, acquired by a Photometrics KAF 1400 camera (Photometrics Ltd., Tuscon, AZ, USA), and then analyzed on a Macintosh Quadra 840AV (Apple Computer, Inc., Cupertino, CA, USA) computer. After the required number of cells has been analyzed, the user may interact to correct the computer by working with a gallery of the cell images. The automated dot counter has been tested on a number of normal specimens where 4,'6-diamidino-2-phenylindole (DAPI) was used for the nucleus counterstain and a centromeric 8 probe was used to mark the desired chromosome. The slides contained lymphocytes from cultured blood. We compared the results of the dot counter with manual counting. Manually obtained results, published in the literature, were used as the "ground truth." For a normal specimen, 97.5% of cells will have two dots. Fully automated scanning of 13 slides showed that an average of 89% of all nuclei were counted correctly. In other words, an average of 11% has to be interactively corrected, using a monitor display. The machine accuracies, after interactive correction, are comparable to panels of human experts (manual). The fully automatically obtained results are biased with respect to manual counting. An error analysis is given, and different causes are discussed. PMID:9136750

Netten, H; Young, I T; van Vliet, L J; Tanke, H J; Vroljik, H; Sloos, W C

1997-05-01

76

Prognostic study of continuous variables (white blood cell count, peripheral blast cell count, haemoglobin level, platelet count and age) in childhood acute lymphoblastic leukaemia. Analysis of a population of 1545 children treated by the French Acute Lymphoblastic Leukaemia Group (FRALLE)  

PubMed Central

Many cutpoints have been proposed to categorize continuous variables in childhood acute lymphoblastic leukaemia (white blood cell count, peripheral blast cell count, haemoglobin level, platelet count and age), and have been used to define therapeutic subgroups. This variation in the choice of cutpoints leads to a bias called the ‘Will Rogers phenomenon’. The aim of this study was to analyse variations in the relative risk of relapse or death as a function of continuous prognostic variables in childhood ALL and to discuss the choice of cutpoints. We studied a population of 1545 children with ALL enrolled in three consecutive protocols named FRALLE 83, FRALLE 87 and FRALLE 89. We estimated the risk of relapse or death associated with different values of each continuous prognostic variable by dividing the sample into quintiles of the distribution of the variables. As regards age, a category of children under 1 year of age was distinguished and the rest of the population was divided into quintiles. The floated variance method was used to calculate the confidence interval of each relative risk, including the reference category. The relation between the quantitative prognostic factors and the risk was monotonic for each variable, except for age. For the white blood cell count (WBC), the relation is log linear. The risk associated with WBC values in the upper quintile was 1.9 times higher than that in the lower quintile. The peripheral blast cell count correlated strongly with WBC (correlation coefficient: 0.99). The risk increased with the haemoglobin level, and the risk in the upper quintile was 1.3 times higher than that in the lower quintile. The risk decreased as the platelet count increased: the risk in the lower quintile was 1.2 times higher than that in the upper quintile. The risk increased gradually with increasing age above one year. The small subgroup of patients (2.5% of the population) under 1 year of age at diagnosis had a risk 2.6 times higher than the reference category of patients between 3 and 4.3 years of age. When the risk associated with a quantitative prognostic factor varies monotonously, the selection of a cutpoint is arbitrary and represents a loss of information. Despite this loss of information, such arbitrary categorization may be necessary to define therapeutic stratification. In that case, consensus cutpoints must be defined if one wants to avoid the Will Rogers phenomenon. The cutpoints proposed by the Rome workshop and the NCI are arbitrary, but may represent an acceptable convention. © 2000 Cancer Research Campaign http://www.bjcancer.com PMID:11104555

Donadieu, J; Auclerc, M-F; Baruchel, A; Perel, Y; Bordigoni, P; Landman-Parker, J; Leblanc, T; Cornu, G; Sommelet, D; Leverger, G; Schaison, G; Hill, C

2000-01-01

77

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

NASA Astrophysics Data System (ADS)

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

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

2011-07-01

78

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

PubMed

This paper presents our method, which simultaneously combines automatic imaging, identification, and counting with the acquisition of morphological information for at least 1000 blood cells from several three-dimensional images of the same sample. We started with seeking parameters to differentiate between red blood cells that are similar but different with respect to their development stage, i.e., mature or immature. We highlight that these cells have different diffractive patterns with complementary central intensity distribution in a given plane along the propagation axis. We use the Fresnel approximation to simulate propagation through cells modeled as spheroid-shaped phase objects and to find the cell property that has the dominant influence on this behavior. Starting with images obtained in the reconstruction step of the digital holographic microscopy technique, we developed a code for automated simultaneous individual cell image separation, identification, and counting, even when the cells are partially overlapped on a slide, and accurate measuring of their morphological features. To find the centroids of each cell, we propose a method based on analytical functions applied at threshold intervals. Our procedure separates the mature from the immature red blood cells and from the white blood cells through a decision based on gradient and radius values. PMID:21743570

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

2011-07-10

79

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

PubMed Central

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

2012-01-01

80

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

PubMed Central

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 between physical activity, inflammation, and lung cancer risk was evaluated in a prospective cohort of 4,831 subjects, 43–86 years of age, in Beaver Dam, Wisconsin. A total physical activity index was created by summing kilocalories per week from sweat-inducing physical activities, city blocks walked, and flights of stairs climbed. Two inflammatory markers, white blood cell 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 to those in the lowest tertile (OR=0.55; 95% CI: 0.35–0.86). Participants with white blood cell counts in the upper tertile (?8×103/?L) were 2.81 (95% CI: 1.58–5.01) times as likely to develop lung cancer as those with counts in the lowest tertile (<6.4×103/?L). 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 white blood cell count. While the potential for residual confounding by smoking could not be eliminated, these data suggest that physical activity and white blood cell count are independent risk factors for lung cancer. PMID:18843014

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

2009-01-01

81

Baseline CD4 Cell Counts of Newly Diagnosed HIV Cases in China: 2006-2012  

PubMed Central

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

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

2014-01-01

82

Fluid mechanics of method of separating motile cells  

NSDL National Science Digital Library

If the Reynolds number is small enough (Re<<1), then two fluids can flow in parallel in direct contact, exchanging momentum and species only by diffusion. If the interface is stable, then this system can be used as a filter. In this problem, the flow fields in both fluids are found. The system here has a diffusing species which is motile cells with a random behavior relative to the flowing fluid.

Krane, Matthew J.; Martinez, Carlos

2008-10-25

83

Neonatal nucleated red blood cell counts in growth-restricted fetuses: Relationship to arterial and venous Doppler studies  

Microsoft Academic Search

Objective: Elevated nucleated red blood cell count in neonatal blood and Doppler-detected circulatory decompensation in fetuses with intrauterine growth restriction are associated with hypoxemia. We sought to determine the relationship between the nucleated red blood cell count at birth and the circulatory status of fetuses with intrauterine growth restriction. Study Design: Eighty-four fetuses with elevated umbilical artery pulsatility index values

Ahmet A. Baschat; Ulrich Gembruch; Irwin Reiss; Ludwig Gortner; Chris R. Harman; Carl P. Weiner

1999-01-01

84

Probing Stemness and Neural Commitment in Human Amniotic Fluid Cells  

Microsoft Academic Search

Recently, human amniotic fluid (AF) cells have attracted a great deal of attention as an alternative cell source for transplantation\\u000a and tissue engineering. AF contains a variety of cell types derived from fetal tissues, of which a small percentage is believed\\u000a to represent stem cell sub-population(s). In contrast to human embryonic stem (ES) cells, AF cells are not subject to

Anna Jezierski; Andree Gruslin; Roger Tremblay; Dao Ly; Cathie Smith; Kursad Turksen; Marianna Sikorska; Mahmud Bani-Yaghoub

2010-01-01

85

Alcohol Consumption and CD4 T-cell count response among persons initiating antiretroviral therapy  

PubMed Central

Background We evaluated the longitudinal association of alcohol use with immunological response to combination antiretroviral therapy (ART) among HIV infected individuals. Methods This was a prospective cohort study of individuals initiating ART. Participants underwent an Audio Computer-Assisted Self Interview querying drug and alcohol use within 6 months of treatment. Immunological response to ART was defined by CD4 T-cell count (CD4). Primary independent variables were self-reported number of drinks consumed per drinking day (quantity) and days of alcohol consumption in a typical week (frequency). We used linear mixed effects models to quantify the association between CD4 T-cell count and alcohol quantity and frequency and Cox proportional hazards models to estimate the relative hazard of an increase 100, 150 and 200 CD4 cells/mm3 per additional drink per drinking day. Analyses were stratified by gender. Viral suppression was examined as a time-varying covariate. Results Between 2000-2008, 1107 individuals were eligible for inclusion in this study. There was no statistically significant difference in CD4 T-cell count by average drinks per drinking day at any frequency of alcohol use irrespective of gender or viral suppression. Similarly, we found no difference in the hazard ratio for drinks per drinking day within the categories of drinking frequency for time to CD4 T-cell count increase of 100, 150 and 200 cells/mm3, respectively. Conclusions Among individuals initiating antiretroviral therapy (ART) the benefits of therapy and viral suppression on the immune system outweigh detrimental effects of alcohol, reinforcing the importance of initiating ART and ensuring adequate adherence to therapy. PMID:22955054

Kowalski, Stefan; Colantuoni, Elizabeth; Lau, Bryan; Keruly, Jeanne; McCaul, Mary E.; Hutton, Heidi E.; Moore, Richard D.; Chander, Geetanjali

2012-01-01

86

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

PubMed Central

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

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

2003-01-01

87

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

PubMed Central

(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/?L 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/?L; 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?cell counts of ?350?cells/?L and percentages of >15%. Study of whether early initiation of ART could prevent or reverse such deficits is needed. PMID:22308272

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

88

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

PubMed

The aim of this study was to describe the temporal variation in bulk milk somatic cell count (BMSCC) on Dutch dairy goat farms and to assess the correlation of BMSCC with bulk milk total bacterial counts (BMTBC) and with several herd management factors. Bulk milk somatic cell count and BMTBC data were recorded from 90% of the dairy goat farms in the Netherlands over the years 2005 to 2007. Farm characteristics and management information was collected by means of questionnaires. The bulk milk data and the questionnaire data were linked and linear mixed models were used to identify risk factors for increased BMSCC and BMTBC. Bulk milk somatic cell count was found to display a distinct pattern throughout the year, being highest around December and lowest around June. Bulk milk somatic cell count correlated to BMTBC (r = 0.4). Significant factors in the BMSCC model were month in lactation, treating mastitic animals instead of culling, caprine arthritis encephalitis status, milk fever prevalence, and liner material. Month in lactation and treating mastitic animals instead of culling were also significant in the BMTBC model. In the high-BMSCC period, a higher number of goats with an extended lactation significantly reduced the BMSCC. Thus, this study indicates that mastitis-related factors account for some of the variation in BMSCC and BMTBC levels between dairy goat herds. It shows that intramammary infection is probably the most important factor driving the correlation between BMSCC and BMTBC, suggesting that programs to improve udder health may have a positive effect on both BMSCC and BMTBC. PMID:19700695

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

2009-09-01

89

Effect of somatic cell count and mastitis pathogens on milk composition in Gyr cows  

PubMed Central

Background Gyr cows are well adapted to tropical conditions, resistant to some tropical diseases and have satisfactory milk production. However, Gyr dairy herds have a high prevalence of subclinical mastitis, which negatively affects their milk yield and composition. The objectives of this study were (i) to evaluate the effects of seasonality, mammary quarter location (rear x front), mastitis-causing pathogen species, and somatic cell count (SCC) on milk composition in Gyr cows with mammary quarters as the experimental units and (ii) to evaluate the effects of seasonality and somatic cell count (SCC) on milk composition in Gyr cows with cows as the experimental units. A total of 221 lactating Gyr cows from three commercial dairy farms were selected for this study. Individual foremilk quarter samples and composite milk samples were collected once a month over one year from all lactating cows for analysis of SCC, milk composition, and bacteriological culture. Results Subclinical mastitis reduced lactose, nonfat solids and total solids content, but no difference was found in the protein and fat content between infected and uninfected quarters. Seasonality influenced milk composition both in mammary quarters and composite milk samples. Nevertheless, there was no effect of mammary quarter position on milk composition. Mastitis-causing pathogens affected protein, lactose, nonfat solids, and total solids content, but not milk fat content. Somatic cell count levels affected milk composition in both mammary quarters and composite samples of milk. Conclusions Intramammary infections in Gyr cows alter milk composition; however, the degree of change depends on the mastitis-causing pathogen. Somatic cell count is negatively associated with reduced lactose and nonfat solids content in milk. Seasonality significantly affects milk composition, in which the concentration of lactose, fat, protein, nonfat solids and total solids differs between dry and wet seasons in Gyr cows. PMID:23566405

2013-01-01

90

A Fluid-Kinetic Particle-in-Cell Solver  

NASA Astrophysics Data System (ADS)

A fluid solver that retains kinetic effects by using the Particle-in-Cell (PIC) algorithm is presented in the context of future coupled fluid-kinetic plasma simulations. The fluid continuity and momentum equations together with the second order formulation of Maxwell's equations are solved concurrently using the finite volume box scheme. The pressure tensor in the fluid momentum equation is self-consistently computed using the computational particles. The electric field is corrected to take into account the discrepancies between the fluid densities calculated from the fluid equation and the one calculated directly from the computational particles. The magnetic field is determined from Faraday's law. Finally, the position and velocity of the computational particles are advanced in time. The fluid-kinetic PIC solver is implemented starting from the iPIC3D code, a massively parallel fully kinetic code. The fluid-kinetic PIC solver method could be used in spatial regions where kinetic effects are important, while a traditional fluid solver would be used in regions of space where the kinetic effects are negligible to save computational time. Therefore, the proposed scheme is a promising approach for coupling fluid and kinetic methods in a unified framework.

Markidis, Stefano; Henri, Pierre; Lapenta, Giovanni; Ronnmark, Kjell; Hamrin, Maria; Laure, Erwin

2012-10-01

91

Spurious platelet counts in acute leukaemia with DIC due to cell fragmentation.  

PubMed

Automated platelet counts in a patient with newly diagnosed AML M5 with extreme leukocytosis were reported as 129, 166 and 121 x 10(9)/1. Routine blood films showed a corresponding number of platelet-sized particles, judged to be platelets. The patient was treated for DIC with low-dose heparin infusion. Platelet transfusions were not given initially. The patient died 14 h after admission from intracerebral haematoma. The origin of the platelet-sized particles seen in routine stained blood films was examined by cytochemical and immunological staining for peroxidase, non-specific esterase, CD 13 and CD 33. About 1/3 of the fragments had the same staining characteristics as the leukaemia cells, indicating leukaemia cell origin. Staining for platelet-specific antigen GpIIIa was positive only in 4% of the platelet-sized fragments, with a calculated true platelet count of 4 x 10(9)/1. The presence of cell fragments masquerading as platelets should be suspected in leukaemia patients with bleeding symptoms and normal or near normal platelet counts. PMID:1451403

Hammerstrøm, J

1992-01-01

92

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

PubMed

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. PMID:21618258

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

2012-08-01

93

Salivary biomass assessed by bioluminescence ATP assay related to (bacterial and somatic) cell counts.  

PubMed

The present work aimed (1) to evaluate ATP content in saliva by the bioluminescent luciferin-luciferase method, (2) to evaluate the relationships between ATP content, bacterial count and epithelial cell numbers in saliva, (3) to study the effect of two different antiseptics (peroxidase system producing hypothiocyanite and chlorhexidine) on the salivary biomass. In 45 young adults, the salivary ATP content ranged from 8 to 1515 nM. Salivary ATP content was significantly and directly correlated to bacterial count and epithelial cell numbers (Spearman-Rank correlation, P< or =0.001). Regression analysis allowed the inference of a mean epithelial cell and bacterial ATP content of 152.7 fg and 8.3 fg per cell, respectively. The salivary ATP content decreased significantly to 38. 8+/-12.3 per cent (mean+/-SEM, N=6) of its initial value after a 30-min incubation in the presence of a peroxidase system producing hypothiocyanite (OSCN(-)). Chlorhexidine (CHX) reduced salivary ATP content to 52.0+/-16.7 per cent. OSCN(-) did not affect the transformed logarithm of bacterial count but CHX reduced it from 7. 02+/-0.26 to 0.52+/-0.33. No effect of OSCN(-) was seen on the ratio of epithelial cell viability while CHX reduced it from 46.7+/-5.1 to 3.9+/-1.1 per cent. It is concluded that the combination of the evaluations of the ATP content and cell numbers in saliva can provide reliable data about the effects of oral antiseptics on salivary biomass. PMID:10814968

Gallez, F; Fadel, M; Scruel, O; Cantraine, F; Courtois, P

2000-06-01

94

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

SciTech Connect

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

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

1995-12-31

95

Correlation of Circulating MMP-9 with White Blood Cell Count in Humans: Effect of Smoking  

PubMed Central

Background Matrix metalloproteinase-9 (MMP-9) is an emerging biomarker for several disease conditions, where white blood cell (WBC) count is also elevated. In this study, we examined the relationship between MMP-9 and WBC levels in apparently healthy smoking and non-smoking human subjects. Methods We conducted a cross-sectional study to assess the relationship of serum MMP-9 with WBC in 383 men and 356 women. Next, we divided the male population (women do not smoke in this population) into three groups: never (n?=?243), current (n?=?76) and former (n?=?64) smokers and compared the group differences in MMP-9 and WBC levels and their correlations within each group. Results Circulating MMP-9 and WBC count are significantly correlated in men (R2?=?0.13, p<0.001) and women (R2?=?0.19, p<0.001). After stratification by smoking status, MMP-9 level was significantly higher in current smokers (mean ± SE; 663.3±43.4 ng/ml), compared to never (529.7±20.6) and former smokers (568±39.3). WBC count was changed in a similar pattern. Meanwhile, the relationship became stronger in current smokers with increased correlation coefficient of r?=?0.45 or R2?=?0.21 (p<0.001) and steeper slope of ß?=?1.16±0.30 (p<0.001) in current smokers, compared to r?=?0.26 or R2?=?0.07 (p<0.001) and ß?=?0.34±0.10 (p<0.001) in never smokers. Conclusions WBC count accounts for 13% and 19% of MMP-9 variance in men and women, respectively. In non-smoking men, WBC count accounts for 7% of MMP-9 variance, but in smoking subjects, it accounts for up to 21% of MMP-9 variance. Thus, we have discovered a previously unrecognized correlation between the circulating MMP-9 and WBC levels in humans. PMID:23825535

Ryan, Kathleen A.; Yu, Daozhan; Shuldiner, Alan R.; Mitchell, Braxton D.; Gong, Da-Wei

2013-01-01

96

White blood cell count and mortality in patients with acute pulmonary embolism.  

PubMed

Although associated with adverse outcomes in other cardiovascular diseases, the prognostic value of an elevated white blood cell (WBC) count, a marker of inflammation and hypercoagulability, is uncertain in patients with pulmonary embolism (PE). We therefore sought to assess the prognostic impact of the WBC in a large, state-wide retrospective cohort of patients with PE. We evaluated 14,228 patient discharges with a primary diagnosis of PE from 186 hospitals in Pennsylvania. We used random-intercept logistic regression to assess the independent association between WBC count levels at the time of presentation and mortality and hospital readmission within 30 days, adjusting for patient and hospital characteristics. Patients with an admission WBC count <5.0, 5.0-7.8, 7.9-9.8, 9.9-12.6, and >12.6 × 10(9) /L had a cumulative 30-day mortality of 10.9%, 6.2%, 5.4%, 8.3%, and 16.3% (P < 0.001), and a readmission rate of 17.6%, 11.9%, 10.9%, 11.5%, and 15.0%, respectively (P < 0.001). Compared with patients with a WBC count 7.9-9.8 × 10(9) /L, adjusted odds of 30-day mortality were significantly greater for patients with a WBC count <5.0 × 10(9) /L (odds ratio [OR] 1.52, 95% confidence interval [CI] 1.14-2.03), 9.9-12.6 × 10(9) /L (OR 1.55, 95% CI 1.26-1.91), or >12.6 × 10(9) /L (OR 2.22, 95% CI 1.83-2.69), respectively. The adjusted odds of readmission were also significantly increased for patients with a WBC count <5.0 × 10(9) /L (OR 1.34, 95% CI 1.07-1.68) or >12.6 × 10(9) /L (OR 1.29, 95% CI 1.10-1.51). In patients presenting with PE, WBC count is an independent predictor of short-term mortality and hospital readmission. PMID:23674436

Venetz, Carmen; Labarère, José; Jiménez, David; Aujesky, Drahomir

2013-08-01

97

[The changes in complete blood count in patients treated with sunitinib malate for metastatic clear cell renal cell carcinoma].  

PubMed

Renal cell carcinoma (RCC) accounts for approximately 3% of adult malignancies. For stage I - III RCC surgery is the primary treatment. Systemic therapy is used in the patients with disseminated disease (stage IV). Sunitinib malate is commonly used in the patients with clear cell renal cell carcinoma (ccRCC) rated as 'low' or 'intermediate' risk according to the Motzer scale. Treatment with sunitinib malate is associated with myelotoxicity. To assess its clinical significance we conducted a pilot study in a group of 10 patients. We noticed a gradual decrease in the mean haemoglobin level during subsequent treatment cycles. Alternations in the platelet count were of no clinical significance. Episodes of the neutropenia were noticed in the study group. In some patients neutrophil count decreased to the level that put them at risk of the infectious complications. PMID:24455830

Kucharz, Jakub; Micha?owska-Kaczmarczyk, Anna; Streb, Joanna; Kuzniewski, Marek; Herman, Roman M; Krzemieniecki, Krzysztof

2013-01-01

98

Fluids as Dynamic Templates for Cytoskeletal Proteins in Plant Cells  

E-print Network

The Dynamic Template model of biological cell membranes and the cytoplasm as spatially organised fluid layers is extended to plant cells, and is shown to offer a feasible shear driven mechanism for the co-alignment of internal and external fibres observed during growth and tropic responses

J. T. Lofthouse

2008-07-12

99

Three counting methods agree on cell and neuron number in chimpanzee primary visual cortex  

PubMed Central

Determining the cellular composition of specific brain regions is crucial to our understanding of the function of neurobiological systems. It is therefore useful to identify the extent to which different methods agree when estimating the same properties of brain circuitry. In this study, we estimated the number of neuronal and non-neuronal cells in the primary visual cortex (area 17 or V1) of both hemispheres from a single chimpanzee. Specifically, we processed samples distributed across V1 of the right hemisphere after cortex was flattened into a sheet using two variations of the isotropic fractionator cell and neuron counting method. We processed the left hemisphere as serial brain slices for stereological investigation. The goal of this study was to evaluate the agreement between these methods in the most direct manner possible by comparing estimates of cell density across one brain region of interest in a single individual. In our hands, these methods produced similar estimates of the total cellular population (approximately 1 billion) as well as the number of neurons (approximately 675 million) in chimpanzee V1, providing evidence that both techniques estimate the same parameters of interest. In addition, our results indicate the strengths of each distinct tissue preparation procedure, highlighting the importance of attention to anatomical detail. In summary, we found that the isotropic fractionator and the stereological optical fractionator produced concordant estimates of the cellular composition of V1, and that this result supports the conclusion that chimpanzees conform to the primate pattern of exceptionally high packing density in V1. Ultimately, our data suggest that investigators can optimize their experimental approach by using any of these counting methods to obtain reliable cell and neuron counts. PMID:24904305

Miller, Daniel J.; Balaram, Pooja; Young, Nicole A.; Kaas, Jon H.

2014-01-01

100

Depression severity is associated with increased risk behaviors and decreased CD4 cell counts.  

PubMed

Depression is a common comorbidity among HIV-infected individuals. We studied the relationship between depressive symptoms, risk behaviors (risky-sexual behavior, tobacco, alcohol, and illicit drug use) and HIV outcomes. This cross-sectional study conducted in 2009 at the Washington University HIV Clinic included screening for depression with patient health questionnaire, survey of sexual behavior, illicit drug, alcohol, and tobacco use within 30 days. Sociodemographics, plasma HIV RNA levels, CD4 cell counts, and sexually transmitted disease test results were obtained from medical records. Multivariate logistic and linear regression models were used to assess the association between depressive symptoms severity and risk behaviors, HIV outcomes and combination antiretroviral therapy (cART) adherence. A total of 624 persons completed the assessment of whom 432 (69%) were male and 426 (68%) African-American. The median CD4 cell count was 410 cells/mm(3) and 479 persons (77%) were on cART of whom 112 (23%) had HIV RNA level > 400 copies/mL. Overall, 96 (15%) had symptoms of major depressive disorder. Depressive symptom severity was associated with increased likelihood of high-risk drinking (odds ratio [OR], 2.4; 95% confidence interval [CI], 1.1-5.1), current tobacco use (OR, 1.8; 95% CI, 1.1-2.9), illicit drug use (OR, 1.7; 95% CI, 1.0-2.8), and risky-sexual behavior (OR, 1.5; 95% CI, 0.8-2.7). Suboptimal cART adherence (visual analog scale < 95%) was also associated with depressive symptoms severity (p < 0.05). After adjustment for age, sex, race, receipt of cART, and cART adherence, depressive symptoms severity was independently associated with lower CD4 cell count (p < 0.05) but not with higher HIV RNA level (p = 0.39). Depression adversely affects HIV-infected individuals, requiring greater effort at utilizing multidisciplinary interventions. PMID:24479743

Taniguchi, Toshibumi; Shacham, Enbal; Onen, Nur Fiona; Grubb, Jessica Rosenbaum; Overton, Edgar Turner

2014-01-01

101

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

PubMed Central

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)-1? 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-1? (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 placebo group. These results demonstrate that supplementation with oligonol for 1 week may enhance the immune function under heat and suggest a potential useful adjunct to chemotherapy in malignant diseases. PMID:24962480

Lee, Jeong Beom; Shin, Young Oh

2014-01-01

102

Evaluating Total Lymphocyte Count as a Surrogate Marker for CD4 Cell Count in the Management of HIV-Infected Patients in Resource-Limited Settings: A Study from China  

PubMed Central

Objective To evaluate the correlation of total lymphocyte count (TLC) and CD4 cell count and the suitability of TLC as a surrogate marker for CD4 cell count of HIV-infected patients in China. Methods Usefulness of TLC as a surrogate marker for a CD4 cell count <350 cells/mm3 for HIV-positive patients in China was evaluated by 977 pairs of TLC and CD4 cell count from 977 outpatients. The result was then validated by a literature review which was conducted on 9 relevant articles. Further investigation using the 977 pairs of TLC and CD4 cell count data was done to determine a TLC threshold for predicting a CD4 cell count <500 cells/mm3. Correlation and receiver operating characteristic (ROC) analysis were performed for both CD4 cell counts, and the sensitivity and specificity were computed. Results Good correlation was noted between TLC and CD4 count (r?=?0.60, 95% CI, 0.56–0.64). TLC obtained a relatively high diagnostic performance (area under ROC curve, 0.80) for predicting a CD4 cell count <350 cells/mm3, with a sensitivity of 0.65 (95% CI, 0.61–0.68) and a specificity of 0.80 (95% CI, 0.75–0.85) at the TLC threshold of 1570 cells/mm3. The literature review suggested that for a CD4 cell count <350 cells/mm3, the optimal TLC threshold was 1500 cells/mm3, which was similar to the figure presented in this observational study. As for predicting a CD4 cell count <500 cells/mm3, TLC obtained a high diagnostic performance (area under ROC curve, 0.82) as well with a sensitivity of 0.70 (95% CI, 0.67–0.73) and a specificity of 0.80 (95% CI, 0.73–0.87). Conclusions When considering the antiretroviral therapy for HIV-infected Chinese individuals, total lymphocyte count can be considered as an inexpensive and easily available surrogate marker for predicting two clinically important thresholds of CD4 count of 350 cells/mm3 and 500 cells/mm3. PMID:23874985

Zou, Ran; Yang, Qiuying; Zhang, Hongwei; Zhang, Tong; Chen, Hui; Wu, Hao

2013-01-01

103

Disseminated histoplasmosis in an HIV patient with CD4 count of 1 cell/µL.  

PubMed

We report a case of a young woman with advanced HIV/AIDS who presented with a short duration of fever and shortness of breath, with no recent travel history or previous hospitalisation, accompanied by non-specific abdominal symptoms and suspicious upper gastrointestinal bleed. Her CD4 count was 1?cell/?L raising the suspicious for various opportunistic aetiologies. The initial suspicion was for pneumocystis pneumonia (PCP) and the patient was treated empirically with antimicrobials. Peripheral smear findings, urinary antigen tests and bronchoalveolar lavage (BAL) were suggestive of disseminated histoplasmosis. PCP was ruled out in BAL. Transabdominal imaging was concerning for periaortic lymphadenopathy raising the suspicion of occult malignancy. Endoscopic evaluation of her digestive tract was unrevealing. Endoscopic ultrasound fine-needle aspiration (EUS-FNA) confirmed histoplasmosis. She received a liposomal amphotericin B for 10?days followed by itraconazole with significant improvement. Her CD4 count was found to be the lowest reported count with a single opportunistic pathogen. PMID:25139916

Tella, Sri Harsha; Abuzaid, Ahmed

2014-01-01

104

Alcohol dependence and CD4 cell count: is there a relationship?  

PubMed

Alcohol and other drugs use seem to be common among people infected with HIV on antiretroviral treatment (ART). Their effects on HIV progression is still in debate. This study aimed to assess the association between alcohol and drug use and an HIV disease progression biomarker (CD4 cell count) among patients on ART. A cross-sectional study was carried out at an HIV treatment center affiliated with Medical School of the University of São Paulo, Brazil. Four hundred and thirty-eight HIV-positive patients on ART were interviewed by trained psychiatrists and psychologists using the following instruments: Structured Clinical Interview for DSM-IV Axis I Disorders (SCID-I), Alcohol Use Disorders Identification Test (AUDIT), 17-item Hamilton Rating Scale for Depression, and the Simplified Medication Adherence Questionnaire (SMAQ). In the previous month, 219 (50%) and 41 (9.3%) patients reported use of alcohol and illicit drugs, respectively. Fifty patients (12.6%) were classified as having harmful alcohol use by AUDIT. According to SCID-I, 80 patients (18.3%) were alcohol abusers, 24 (5.5%) alcohol dependents, and 21 (4.2%) had a current depressive disorder. Almost 73% (n = 319-72.8%) of the patients were adherent to ART. Alcohol dependents were nine times (p < 0.01) more likely to have CD4 cell count ?200/mm(3), and this association was independent of ART adherence. In conclusion, alcohol dependence seems to be associated with low CD4 cell count in HIV-positive patients. Based on these data, HIV health care workers should always assess alcohol consumption in the treatment setting, and patients should be advised that alcohol dependence may be linked to low CD4. PMID:25179410

Malbergier, Andre; Amaral, Ricardo Abrantes do; Cardoso, Luciana Donola

2015-01-01

105

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

NASA Technical Reports Server (NTRS)

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 or 2 days), but also in the very late phase (up to 4 weeks) after exposure.

Hu, Shaowen

2014-01-01

106

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

PubMed Central

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

2013-01-01

107

Telomere Length in Elderly Caucasians Weakly Correlates with Blood Cell Counts  

PubMed Central

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

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

2013-01-01

108

Mechanotransduction of fluid stresses governs 3D cell migration  

PubMed Central

Solid tumors are characterized by high interstitial fluid pressure, which drives fluid efflux from the tumor core. Tumor-associated interstitial flow (IF) at a rate of ?3 µm/s has been shown to induce cell migration in the upstream direction (rheotaxis). However, the molecular biophysical mechanism that underlies upstream cell polarization and rheotaxis remains unclear. We developed a microfluidic platform to investigate the effects of IF fluid stresses imparted on cells embedded within a collagen type I hydrogel, and we demonstrate that IF stresses result in a transcellular gradient in ?1-integrin activation with vinculin, focal adhesion kinase (FAK), FAKPY397, F actin, and paxillin-dependent protrusion formation localizing to the upstream side of the cell, where matrix adhesions are under maximum tension. This previously unknown mechanism is the result of a force balance between fluid drag on the cell and matrix adhesion tension and is therefore a fundamental, but previously unknown, stimulus for directing cell movement within porous extracellular matrix. PMID:24550267

Polacheck, William J.; German, Alexandra E.; Mammoto, Akiko; Ingber, Donald E.; Kamm, Roger D.

2014-01-01

109

Amniotic Fluid and Amniotic Membrane Stem Cells: Marker Discovery  

PubMed Central

Amniotic fluid (AF) and amniotic membrane (AM) have been recently characterized as promising sources of stem or progenitor cells. Both not only contain subpopulations with stem cell characteristics resembling to adult stem cells, such as mesenchymal stem cells, but also exhibit some embryonic stem cell properties like (i) expression of pluripotency markers, (ii) high expansion in vitro, or (iii) multilineage differentiation capacity. Recent efforts have been focused on the isolation and the detailed characterization of these stem cell types. However, variations in their phenotype, their heterogeneity described by different groups, and the absence of a single marker expressed only in these cells may prevent the isolation of a pure homogeneous stem cell population from these sources and their potential use of these cells in therapeutic applications. In this paper, we aim to summarize the recent progress in marker discovery for stem cells derived from fetal sources such as AF and AM, using novel methodologies based on transcriptomics, proteomics, or secretome analyses. PMID:22701492

Roubelakis, Maria G.; Trohatou, Ourania; Anagnou, Nicholas P.

2012-01-01

110

Probing stemness and neural commitment in human amniotic fluid cells.  

PubMed

Recently, human amniotic fluid (AF) cells have attracted a great deal of attention as an alternative cell source for transplantation and tissue engineering. AF contains a variety of cell types derived from fetal tissues, of which a small percentage is believed to represent stem cell sub-population(s). In contrast to human embryonic stem (ES) cells, AF cells are not subject to extensive legal or ethical considerations; nor are they limited by lineage commitment characteristic of adult stem cells. However, to become therapeutically valuable, better protocols for the isolation of AF stem cell sub-populations need to be developed. This study was designed to examine the molecular components involved in self-renewal, neural commitment and differentiation of AF cells obtained at different gestational ages. Our results showed that, although morphologically heterogeneous, AF cells derived from early gestational periods ubiquitously expressed KERATIN 8 (K8), suggesting that the majority of these cells may have an epithelial origin. In addition, AF cells expressed various components of NOTCH signaling (ligands, receptors and target genes), a pathway involved in stem cell maintenance, determination and differentiation. A sub-population of K8 positive cells (<10%) co-expressed NESTIN, a marker detected in the neuroepithelium, neural stem cells and neural progenitors. Throughout the gestational periods, a much smaller AF cell sub-population (<1%) expressed pluripotency markers, OCT4a, NANOG and SOX2, from which SOX2 positive AF cells could be isolated through single cell cloning. The SOX2 expressing AF clones showed the capacity to give rise to a neuron-like phenotype in culture, expressing neuronal markers such as MAP2, NFL and NSE. Taken together, our findings demonstrated the presence of fetal cells with stem cell characteristics in the amniotic fluid, highlighting the need for further research on their biology and clinical applications. PMID:20221716

Jezierski, Anna; Gruslin, Andree; Tremblay, Roger; Ly, Dao; Smith, Cathie; Turksen, Kursad; Sikorska, Marianna; Bani-Yaghoub, Mahmud

2010-06-01

111

Fluid dynamics and noise in bacterial cell-cell and cell-surface scattering  

E-print Network

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

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

2011-07-12

112

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

PubMed

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

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

2014-08-01

113

Alternative experiments using the geophysical fluid flow cell  

NASA Technical Reports Server (NTRS)

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.

Hart, J. E.

1984-01-01

114

Behaviour of cell cultures from human amniotic fluid  

Microsoft Academic Search

The growth pattern of cell cultures originating from 11 amniotic fluid specimens have been observed. From each specimen 2 to 12 primary cultures were set up. In most cases growth started simultaneously in the primary cultures originating from one sample. The primary cultures lasted from 7 to 30 days. A variation was found both between cultures from different pregnancies as

L Hasholt

1976-01-01

115

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

E-print Network

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

Kuhl, Tonya L.

116

Evaluation of an easy and affordable flow cytometer for volumetric haematopoietic stem cell counting  

PubMed Central

Background Accurate estimation of haematopoietic stem cell (HSC) counts by flow cytometry may be difficult in laboratories in which sophisticated equipment and staff with specific expertise are not available. Affordable flow cytometers that can perform basic functions may help to overcome these difficulties. In this study we compared HSC and leucocyte counts determined by volumetric and bead-based protocols performed with the small, low-cost Accuri® C6, with those obtained with two gold-standard instruments, the four-colour FACSCalibur® and the eight-colour FACSCantoII®, our reference flow cytometers. Materials and methods With the three cytometers we tested, in parallel, 111 consecutive samples from cord blood, peripheral blood from patients with myelofibrosis and myeloproliferative syndromes, fresh and thawed HSC collected by apheresis and bone marrow products. The findings were compared with one-way ANOVA, Bland-Altman analysis and linear regression. Results The results of HSC and leucocyte enumeration by the three devices were strongly correlated (r2>0.99; p<0.0001). ANOVA performed on different subgroups of samples did not reveal significant differences between HSC count determined by the C6 bead-based and reference flow cytometers in any of the subgroups. Regarding the C6 volumetric protocol, a statistically significant difference was observed only in the cord blood subgroup. Time for instrument set-up, calibration and analysis was slightly longer with Accuri® C6 (40 min) than with FACSCantoII® (30 min). Discussion Accuri® C6 is a reliable instrument for HSC enumeration in fresh samples, using both volumetric and bead-based approaches, although the volumetric protocol on cord blood samples needs to be improved. The Accuri® C6 is easy to use, does not require profound knowledge of flow cytometry and could be employed in an urgent setting. Its performance may be improved by more efficient calibration and shorter analysis time. PMID:24887218

Mariani, Mariagabriella; Colombo, Federico; Assennato, Sonny M.; Frugoni, Cecilia; Cattaneo, Alessandra; Trombetta, Elena; Rebulla, Paolo; Porretti, Laura

2014-01-01

117

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

PubMed

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 SCC. The decrease in antibiotic use by drying off quarters without DCT was not compensated by an increase in antibiotic use for treating CM. Total antibiotic use related to mastitis was reduced by 85% in these quarters. PMID:24746132

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

2014-06-01

118

Relation of an elevated white blood cell count after percutaneous coronary intervention to long-term mortality  

Microsoft Academic Search

Increased inflammatory markers are associated with a poor prognosis after percutaneous coronary intervention. Leukocytes play a key role in inflammation, and an increase in white blood cell (WBC) counts is a nonspecific marker of inflammation. In patients undergoing percutaneous coronary intervention, baseline WBC counts independently predict long-term mortality. In a pooled cohort of patients from the Evaluation of c7E3 for

Vivek Rajagopal; Hitinder S Gurm; Deepak L Bhatt; A. Michael Lincoff; James E Tcheng; Dean J Kereiakes; Neal S Kleiman; Gang Jia; Eric J Topol

2004-01-01

119

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

PubMed Central

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

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

2014-01-01

120

Microscopic differential cell counting to identify inflammatory reactions in dairy cow quarter milk samples.  

PubMed

The diagnosis of intramammary infections is mostly based on somatic cell count (SCC) and bacteriological analysis. As an alternative, differential cell counting (DCC) could be a useful method, because it identifies changes in the relative cell populations before the increase in total cell number occurs. The aim of the study was to identify cytological parameters that could be used in the field to classify mammary quarters as healthy or diseased, comparing cyto-bacteriological results with DCC. Overall, 48 cows were randomly selected from 3 herds in Lombardy region of Italy. Herd A was characterized by the absence of contagious microorganisms; in herds B and C, the prevalence of Staphylococcus aureus was 20 and 50%, respectively. Foremilk samples were aseptically collected from 188 quarters and submitted to bacteriological analysis, SCC, and DCC. For statistical analysis, the samples were clustered into 4 health groups, and DCC results were compared in each group. Ninety-six samples were classified as normal secretions (N), 30 as mastitis (M), 15 as latent mastitis (LM), and 47 as unspecific mastitis (UM) based on SCC and bacteriological results. Single percentages of lymphocytes, polymorphonuclear neutrophilic leukocytes (PMNL), or macrophages were first evaluated to established variables capable of identifying healthy and inflamed quarters. Then, combinations of cell populations were tested to increase the discrimination power of DCC: phagocytes, logarithmic PMNL:lymphocyte ratio, and logarithmic phagocyte:lymphocyte ratio. The mean percentage of lymphocytes was significantly higher in group N than in groups LM, UM, and M. The mean percentage of PMNL was significantly lower in group N than in groups UM and M, but not LM. Mean percentages of macrophages were not significantly influenced by the 4 groups. The mean value of phagocytes was significantly lower in group N than in the other groups. Both the logarithmic PMNL:lymphocyte and the logarithmic phagocyte:lymphocyte ratios were significantly lower in group N than in groups LM, UM, and M. Fisher (F-)values were calculated, and the highest F-value was that of log PMNL:lymphocytes ratio (48.23). The explanation for this could be that log PMNL:Lym is the only variable that involved both cell populations statistically influenced by health groups but excluded macrophages. Microscopic DCC has potential as a tool to identify cows affected by any inflammatory process of the mammary gland, with the best results being achieved using log PMNL:lymphocyte as variable. PMID:22818454

Pilla, R; Schwarz, D; König, S; Piccinini, R

2012-08-01

121

Higher somatic cells counted by the electronic counter method do not influence renneting properties of goat milk  

Microsoft Academic Search

Laws of different countries regarding SCC of goat milk are not in agreement with each other and sometimes they fix a threshold for the enhancement of dairy products. The aim of this study was to assess if renneting properties of goat milk are influenced by higher somatic cell count (SCC) measured by an electronic cell counter. Milk samples, taken throughout

M. Pazzola; F. Balia; V. Carcangiu; M. L. Dettori; G. Piras; G. M. Vacca

122

Who is winning the cell-phone wars? Answer: it depends how you count. Units, market share, revenue, profit.  

E-print Network

Who is winning the cell-phone wars? Answer: it depends how you count. Units, market share, revenue quarter, not that far off a million a day. iPhone sales increased by 4 million but its market share launched. According to another analyst, Cannaccord, Apple now has four percent of the cell-phone market

South Bohemia, University of

123

Cerebrospinal fluid supports viability and proliferation of cortical cells in vitro, mirroring in vivo development  

Microsoft Academic Search

BACKGROUND: The central nervous system develops around a fluid filled compartment. Recently, attention has turned to the potential role of the fluid (cerebrospinal fluid, CSF) in the developmental process. In particular, the cerebral cortex develops from the germinal epithelium adjacent to the CSF with regulation of cell proliferation and differentiation provided by cells adjacent to the fluid-filled subarachnoid space. METHODS:

Jaleel A Miyan; Mahjiub Zendah; Farhad Mashayekhi; P Jane Owen-Lynch

2006-01-01

124

Laboratory adverse events and discontinuation of therapy according to CD4+ cell count at the start of antiretroviral therapy  

PubMed Central

Objective: Few data describe antiretroviral treatment (ART)-related adverse events when treatment is initiated at CD4+ cell counts more than 350?cells/?l. We compared rates of laboratory-defined adverse events (LDAEs) according to CD4+ cell count at ART initiation. Design: Analysis of on-going cohort study. Methods: ART-naive persons initiating ART from 2000 to 2010 were included. Chi-square, analysis of variance (ANOVA) and Kruskal–Wallis tests compared characteristics among those starting ART with a CD4+ cell count of 350 or less, 351–499 and at least 500?cells/?l. Time-updated Poisson regression compared rates of LDAE in the three CD4+ cell strata. Cox proportional hazard models compared risk of ART discontinuation. Results: Nine thousand, four hundred and six individuals were included: median age 37 years, 61% white, 80% men, median viral load 4.8?log copies/ml. Four hundred and forty-seven (4.9%) and 1099 (11.7%) started ART with a CD4+ cell count at least 500 and 351–499?cells/?l, respectively. One thousand, two hundred and eighty-three (13.6%) patients experienced at least one LDAE. The rate of LDAE did not differ between those starting ART with a CD4+ cell count 351–499 and less than 350?cells/?l [relative rate 0.90, 95% confidence interval (CI) 0.74–1.09)], but an increased risk of ART discontinuation was observed (hazard ratio 1.58, 95% CI 1.10–2.27). Those starting ART at CD4+ cell count at least 500?cells/?l had an increased rate of LDAE (relative rate 1.44, 95% CI 1.13–1.82) but were not more likely to discontinue ART (hazard ratio 1.15, 95% CI 0.64–2.09). Conclusion: This study demonstrates the need to consider ART-related toxicities when initiating therapy at CD4+ cell counts at least 500?cells/?l. Whilst evidence from randomized controlled trials is awaited, the timing of ART initiation in terms of benefits and risks of ART remains an important question. PMID:24583670

Jose, Sophie; Quinn, Killian; Hill, Teresa; Leen, Clifford; Walsh, John; Hay, Phillip; Fisher, Martin; Post, Frank; Nelson, Mark; Gompels, Mark; Johnson, Margaret; Chadwick, David; Gilson, Richard; Sabin, Caroline; Fidler, Sarah

2014-01-01

125

Crazy Counting  

NSDL National Science Digital Library

Let's count shapes and animals! Let's go to the farm to do some Apple Counting. Then let's have some Counting Fun with shapes and animals. Afterward let's grab a fishing pole and do some Fishy Counting! ...

Terch, Ms.

2010-01-27

126

Human Amniotic Fluid Stem Cell Preconditioning Improves Their Regenerative Potential  

PubMed Central

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

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

127

CHANGES IN NUMBERS AND HEPARIN CONTENT OF PERITONEAL FLUID MAST CELLS OF GROWING RATS MEASURED BY FLOW CYTOFLUOROMETRY  

Microsoft Academic Search

A cytofluorometric method, based on berberine staining of mast cell heparmn, was used for flow cytofluorometric counting and heparin quantitation of mast cells in crude peritoneal suspensions of growing rats. The automatic flow cytofluorometric counting of mast cells correlated well with hemocytometer cell counts. The mean mast cell heparin content obtained by flow cytofluorometry showed good agreement with such obtained

GOSTA BERLIN; LENNART ENERBACK

1978-01-01

128

Amniotic Fluid-Derived Stem Cells for Cardiovascular Tissue Engineering Applications  

E-print Network

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

Petsche Connell, Jennifer

129

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

PubMed

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. PMID:22981577

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

2012-11-01

130

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

PubMed Central

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

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

2014-01-01

131

In vitro and in vivo cardiomyogenic differentiation of amniotic fluid stem cells.  

PubMed

Cell therapy has developed as a complementary treatment for myocardial regeneration. While both autologous and allogeneic uses have been advocated, the ideal candidate has not been identified yet. Amniotic fluid-derived stem (AFS) cells are potentially a promising resource for cell therapy and tissue engineering of myocardial injuries. However, no information is available regarding their use in an allogeneic context. c-kit-sorted, GFP-positive rat AFS (GFP-rAFS) cells and neonatal rat cardiomyocytes (rCMs) were characterized by cytocentrifugation and flow cytometry for the expression of mesenchymal, embryonic and cell lineage-specific antigens. The activation of the myocardial gene program in GFP-rAFS cells was induced by co-culture with rCMs. The stem cell differentiation was evaluated using immunofluorescence, RT-PCR and single cell electrophysiology. The in vivo potential of Endorem-labeled GFP-rAFS cells for myocardial repair was studied by transplantation in the heart of animals with ischemia/reperfusion injury (I/R), monitored by magnetic resonance imaging (MRI). Three weeks after injection a small number of GFP-rAFS cells acquired an endothelial or smooth muscle phenotype and to a lesser extent CMs. Despite the low GFP-rAFS cells count in the heart, there was still an improvement of ejection fraction as measured by MRI. rAFS cells have the in vitro propensity to acquire a cardiomyogenic phenotype and to preserve cardiac function, even if their potential may be limited by poor survival in an allogeneic setting. PMID:21120638

Bollini, Sveva; Pozzobon, Michela; Nobles, Muriel; Riegler, Johannes; Dong, Xuebin; Piccoli, Martina; Chiavegato, Angela; Price, Anthony N; Ghionzoli, Marco; Cheung, King K; Cabrelle, Anna; O'Mahoney, Paul R; Cozzi, Emanuele; Sartore, Saverio; Tinker, Andrew; Lythgoe, Mark F; De Coppi, Paolo

2011-06-01

132

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

NASA Astrophysics Data System (ADS)

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.

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

2014-02-01

133

An adaptive cut-cell method for environmental fluid mechanics  

Microsoft Academic Search

SUMMARY In this work we present a numerical method for solving the incompressible Navier-Stokes equations in an environmental fluid mechanics context. The method is designed for the study of environmental flows that are multiscale, incompressible, variable-density, and within arbitrarily complex and possibly anisotropic domains. The method is new because in this context we couple the embedded-boundary (or cut-cell) method for

Michael F. Barad; Phillip Colella; S. Geoffrey Schladow

2009-01-01

134

Computational fluid dynamics modeling of proton exchange membrane fuel cells  

Microsoft Academic Search

A transient, multi-dimensional model has been developed to simulate proton exchange membrane (PEM) fuel cells. The model accounts simultaneously for electrochemical kinetics, current distribution, hydrodynamics and multi-component transport. A single set of conservation equations valid for flow channels, gas-diffusion electrodes, catalyst layers and the membrane region are developed and numerically solved using a finite-volume-based computational fluid dynamics (CFD) technique. The

SUKKEE UM; C.-Y. Wang; KEN S. CHEN

2000-01-01

135

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

PubMed Central

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

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

2012-01-01

136

Fluid and Cell Transport Through a Microfabricated Flow Chamber.  

NASA Astrophysics Data System (ADS)

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

Brody, James Patrick

137

In Vitro Cardiomyogenic Potential of Human Amniotic Fluid Stem Cells  

PubMed Central

Stem cell therapy for damaged cardiac tissue is currently limited by a number of factors, including the inability to obtain sufficient cell numbers, the potential tumorigenicity of certain types of stem cells, and the possible link between stem cell therapy and the development of malignant arrhythmias. In this study, we investigated whether human amniotic fluid-derived stem (hAFS) cells could be a potential source of cells for cardiac cell therapy by testing the in vitro differentiation capabilities. Undifferentiated hAFS cells express several cardiac genes, including the transcription factor mef2, the gap junction connexin43, and H- and N-cadherin. A 24-hour incubation with 5-aza-2?–deoxycytidine (5-AZA-dC) induced hAFS cell differentiation along the cardiac lineage. Evidence for this differentiation included morphological changes, up-regulation of cardiac-specific genes (cardiac troponin I and cardiac troponin T) and redistribution of connexin43, as well as down-regulation of the stem cell marker SRY-box 2 (sox2). When co-cultured with neonatal rat cardiomyocytes (NRCs), hAFS cells formed both mechanical and electrical connections with the NRCs. Dye transfer experiments showed that calcein dye could be transferred from NRCs to hAFS cells through cellular connections. The gap junction connexin 43 likely involved in the communication between the two cell types, because 12-O-Tetradecanoylphorbol 13-acetate (TPA) could partially block cellular crosstalk. We conclude that hAFS cells can be differentiated into a cardiomyocyte-like phenotype and can establish functional communication with NRCs. Thus, hAFS cells may potentially be used for cardiac cell therapy. PMID:20687122

Guan, Xuan; Delo, Dawn M.; Atala, Anthony; Soker, Shay

2010-01-01

138

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

SciTech Connect

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

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

2011-12-16

139

Estimated retinal ganglion cell counts in glaucomatous eyes with localized retinal nerve fiber layer defects  

PubMed Central

Purpose To estimate retinal ganglion cell (RGC) losses associated with visible glaucomatous localized retinal nerve fiber layer (RNFL) defects. Design Observational cross-sectional study. Methods A multicenter study of 198 normal eyes (138 subjects) and 66 glaucomatous eyes (55 subjects) recruited from the Diagnostic Innovations in Glaucoma Study and African Descent and Glaucoma Evaluation Study. All eyes had standard automated perimetry (SAP), spectral domain optical coherence tomography (SD-OCT) and fundus stereophotographs within 6 months. Glaucomatous eyes were included if localized RNFL defects were detected by masked grading of stereophotographs. The number of RGCs in each sector of a structure-function map was estimated using a previously published model combining RGC estimates from SAP and SD-OCT. The estimated percentage loss of RGCs (combined structure function index) was calculated. Results In glaucomatous eyes there were 136 sectors with visible RNFL defects and 524 sectors without visible RNFL defects. The commonest sectors with visible RNFL defects were inferior and inferotemporal sectors, followed by superior and superotemporal sectors. Eyes with visible RNFL defects had a mean estimated RGC count of 657,172 cells versus 968,883 cells in healthy eyes (P<0.001). The average combined structure function index in sectors with a visible RNFL defect (59±21%) was significantly higher than in sectors without a visible RNFL defect in glaucomatous eyes (15±29%, P<0.001) and higher than in healthy eyes (1±13%, P<0.001). Conclusions Although visible localized RNFL defects are often considered an early sign of glaucoma this study indicates that they are likely to be associated with large neuronal losses. PMID:23746612

Tatham, Andrew J.; Weinreb, Robert N.; Zangwill, Linda M.; Liebmann, Jeffrey M.; Girkin, Christopher A.; Medeiros, Felipe A.

2013-01-01

140

Fred Hutchinson Cancer Research Center study examines predicting ovarian cancer by counting tumor-attacking immune cells  

Cancer.gov

Scientists at Fred Hutchinson Cancer Research Center have developed a new method for counting a special class of cancer-fighting cells – called tumor-infiltrating T lymphocytes, or TILs – reliably, quickly and cheaply in patients with early stage and advanced ovarian cancer.

141

Association Between Somatic Cell Count in Early Lactation and Culling of Dairy Heifers Using Cox Frailty Models  

Microsoft Academic Search

The association between somatic cell count (SCC) of dairy heifers in early lactation (SCCel; measured be- tween 5 and 14 d in milk (DIM)) and the culling hazard during the first lactation was studied using Cox frailty models. Udder health problems were the culling reason for 10% of the culled heifers in this study. For each unit increase in the

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

2005-01-01

142

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

E-print Network

in a dialysis process to filter HIV-1 from the blood5,6 . In these tests, affinity hemodialysis captured gp120Modeling HIV-1 Dynamics and the Effects of Decreasing Activated Infected T-cell Count by Filtration years mathematical models have been developed using differential equations for the progression of HIV-1

Stryker, Gabrielle A.

143

Neuronal characteristics of amniotic fluid derived cells after adenoviral transformation.  

PubMed

Efficient transformation of primary human amniocytes by E1 gene functions of human adenovirus serotype 5 (Ad5) yield in stable cell lines, which exhibit morphological features of epithelial like cells. A thorough investigation using immunocytochemistry confirmed the expression of epithelial cell markers. The analysis also revealed the expression of neuronal and glial marker proteins, such as nestin, vimentin, A2B5 and GFAP. Using RT-PCR, transcripts of the neurotrophic factors nerve growth factor (NGF), brain derived neurotrophic factor (BDNF), glial cell line derived neurotrophic factor (GDNF), and neurotrophin 3 (NT-3) could be detected. Neurotrophic factors could also be detected in the cell culture supernatants of transformed amniocytes. In line with previous experimental data on a human Ad5 E1-transformed embryonal kidney cell line (HEK-293), the results suggest a co-expression of epithelial and neuronal marker proteins in E1-transformed human amniotic fluid derived cells and thus a preferential transformation into neuronal-like cells. PMID:18852058

Arnhold, S; Post, C; Glüer, S; Hoopmann, M; Wenisch, S; Volpers, C; Addicks, K

2008-12-01

144

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

PubMed

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. PMID:22531932

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

2012-07-01

145

Microparticle and cell counting with digital microfluidic compact disc using standard CD drive.  

PubMed

Lab-on-chip medical diagnostics in a global health setting would greatly benefit from highly portable, cost effective and readily available devices. Digital compact disc (CD) and the corresponding detection device-CD drives-for personal computers are extremely affordable and distributable worldwide, therefore they can be immediately used in global health applications if empowered with molecular and cellular biosensing functions. Here we present a novel digital microfluidic CD device derived from conventional music or data CD and demonstrate its preliminary application of counting polystyrene microparticles and living cells in minute-volume fluidic samples. No other detection instruments except for a standard CD drive in a personal computer is used for reading and decoding the quantitative liquid sample information from the digital microfluidic CD. The results presented herein are the first step towards creating a truly portable, low-cost and ubiquitously accessible device-health diagnostic compact disc (HDCD)-for biosensing and health diagnostics, especially in remote or impoverished settings with limited medical infrastructure and healthcare workers. PMID:21350788

Imaad, Syed M; Lord, Nathan; Kulsharova, Gulsim; Liu, Gang Logan

2011-04-21

146

Thermodynamic and fluid properties of cells, tissues and membranes  

NASA Astrophysics Data System (ADS)

This dissertation studies cellular rearrangements in tissues and attempts to establish the role of physical properties of cells, tissues and membranes in several biological phenomena. Using experiments and statistical mechanical modeling, we study cell sorting, tissue engulfment, single cell motion and membrane fluctuations. When cells of two different types are mixed together, they sort out, with the less cohesive tissue surrounding the more cohesive one. This sorting out resembles the phase separation of a mixture of immiscible liquids. We have measured the rate of sorting in tissues and compared it with a cellular automaton based model of cell aggregates. We have also established that cell sorting agrees well with the theory for phase separating fluids. Engulfment is the spreading of one type of tissue over the surface of another tissue placed adjacent to it. Differences in adhesion cause an imbalance of surface tension forces which drives tissue spreading. We have quantitatively studied engulfment between different tissue types and compared the experimental rate with results from computer simulations and a liquid model. Our results suggest that simple physical principles can model tissue motion. Studying the motion of single cells in aggregates is important to understanding the overall pattern formation in tissues. We characterized cell motion in different types of adhesive aggregates to elucidate the role of adhesion in cell motion. We also observed that the cells exhibited a novel type of statistics including correlations and collective motion. Membrane deformations of cells played a negligible role in large scale cell motion. Our results indicate the importance of correlated motion for cells to move long distances in tissues. At the single cell level, tension of the cell membrane and intracellular membrane can play an important role in cell shape changes, regulation of cell motility and membrane dynamics. We used optical tweezers to measure the membrane tension of tubulo-vesicular networks obtained from Golgi and Endoplasmic Reticulum (ER) membranes within cells. As expected on the basis of some previous experiments, the ER has a higher membrane tension than the Golgi.

Upadhyaya, Arpita

2000-10-01

147

Multipotency of equine mesenchymal stem cells derived from synovial fluid.  

PubMed

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

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

2014-10-01

148

Graphite Nodule and Eutectic Cell Count in Cast Iron: Theoretical Model Based on Weibull Statistics and Experimental Verification  

NASA Astrophysics Data System (ADS)

In this work, a model is proposed for heterogeneous nucleation on substrates whose size distribution can be described by the Weibull statistics. It is found that the nuclei density, N nuc can be given in terms of the maximum undercooling, ?T m , by N nuc = N s exp ( b/?T m ), where N s is the density of nucleation sites in the melt and b is the nucleation coefficient (b > 0). When nucleation occurs on all possible substrates, the graphite nodule density, N V,n , or eutectic cell density, N V , after solidification equals N s . In this work, measurements of N V,n and N V values were carried out on experimental nodular and flake graphite iron castings processed under various inoculation conditions. The volumetric nodule N V,n or graphite eutectic cell N V count was estimated from the area nodule count, N A,n , or eutectic cell count, N A , on polished cast iron surface sections by stereological means. In addition, maximum undercoolings, ?T m , were measured using thermal analysis. The experimental outcome indicates that the N V,n or N V count can be properly described by the proposed expression N V,n = N V = N s exp ( b/?T m ). Moreover, the N s and b values were experimentally determined. In particular, the proposed model suggests that the size distribution of nucleation sites is exponential in nature.

Fra?, E.; Wiencek, K.; Górny, M.; López, H. F.

2007-02-01

149

Coupling EELS/EFTEM Imaging with Environmental Fluid Cell Microscopy  

SciTech Connect

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

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

2012-01-01

150

Estimating malaria parasite density among pregnant women at central Sudan using actual and assumed white blood cell count  

PubMed Central

Background Microscopic examination using Giemsa-stained thick blood films remains the reference standard for detection of malaria parasites and it is the only method that is widely and practically available for quantifying malaria parasite density. There are few published data (there was no study during pregnancy) investigating the parasite density (ratio of counted parasites within a given number of microscopic fields against counted white blood cells (WBCs) using actual number of WBCs. Methods Parasitaemia was estimated using assumed WBCs (8,000), which was compared to parasitaemia calculated based on each woman’s WBCs in 98 pregnant women with uncomplicated Plasmodium falciparum malaria at Medani Maternity Hospital, Central Sudan. Results The geometric mean (SD) of the parasite count was 12,014.6 (9,766.5) and 7,870.8 (19,168.8) ring trophozoites /?l, P <0.001 using the actual and assumed (8,000) WBC count, respectively. The median (range) of the ratio between the two parasitaemias (using assumed/actual WBCs) was 1.5 (0.6-5), i e, parasitaemia calculated assuming WBCs equal to median (range) 1.5 (0.6-5) times higher than parasitaemia calculated using actual WBCs. There were 52 out of 98 patients (53%) with ratio between 0.5 and 1.5. For 21 patients (21%) this ratio was higher than 2, and for five patients (5%) it was higher than 3. Conclusion The estimated parasite density using actual WBC counts was significantly lower than the parasite density estimated using assumed WBC counts. Therefore, it is recommended to use the patient`s actual WBC count in the estimation of the parasite density. PMID:24386962

2014-01-01

151

Fluids  

NSDL National Science Digital Library

This Topic in Depth explores the Web's offerings on the physics of fluids. By an educational Web site called School for Champions, the first site is the Fluids lesson plan (1). Here, students or anyone interested can read about the basics of fluids and then take a short interactive quiz on the topic. The second site is maintained by Steve Lower of the Department of Chemistry at Simon Fraser University called Liquids and their Vapors (2). This Adobe Acrobat (.pdf) file contains an eighteen-page document that covers topics such as properties of liquids and changes of state. The next site contains an interactive multimedia activity presented by explorescience.com called Floating Log (3). The site allows users to explore how a fluid can affect buoyancy by letting them change the mass of the log and the fluid's density. The next site from Purdue University's Chemical Education Web site is called Liquids (4). This page describes the structure of liquids, what kinds of materials form liquids, vapor pressure, and more. The fifth site, offered by Professor M.S. Cramer at the College of Engineering at Virginia Tech, is entitled Gallery of Fluid Dynamics (5). It contains movies, animations, photographs, and descriptions of various fluid mechanics topics such as condensation, shock waves, and supersonic cars. Next comes the Innovative Technology Solutions Corporation's Fundamental Fluid Mechanics Movies Web site (6). Over thirty short films show how fluids move in various conditions including gravity waves, fire, material transport, and hydraulics. From the University of Waterloo's Department of Mechanical Engineering-Microelectronics Heat Transfer Laboratory comes the next site, called the Fluid Properties Calculator (7). This online tool allows users to select a fluid and enter a temperature to calculate various parameters such as density, viscosity, specific heat, and thermal diffusivity. The last site is the online journal Physics of Fluids (8), which is published monthly by the American Institute of Physics with the cooperation of The American Physical Society Division of Fluid Dynamics. The journal is "devoted to the publication of original theoretical, computational, and experimental contributions to the dynamics of gases, liquids, and complex or multiphase fluids" and provides free full-text articles for online viewing.

Brieske, Joel A.

2002-01-01

152

A cell-based sensor of fluid shear stress for microfluidics  

E-print Network

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

Varma, Sarvesh

2013-01-01

153

Stem Cells Derived from Human Amniotic Fluid Contribute to Acute Kidney Injury Recovery  

PubMed Central

Stem cells isolated from human amniotic fluid are gaining attention with regard to their therapeutic potential. In this work, we investigated whether these cells contribute to tubular regeneration after experimental acute kidney injury. Cells expressing stem cell markers with multidifferentiative potential were isolated from human amniotic fluid. The regenerative potential of human amniotic fluid stem cells was compared with that of bone marrow-derived human mesenchymal stem cells. We found that the intravenous injection of 3.5 × 105 human amniotic fluid stem cells into nonimmune-competent mice with glycerol-induced acute kidney injury was followed by rapid normalization of renal function compared with injection of mesenchymal stem cells. Both stem cell types showed enhanced tubular cell proliferation and reduced apoptosis. Mesenchymal stem cells were more efficient in inducing proliferation than amniotic fluid-derived stem cells, which, in contrast, were more antiapoptotic. Both cell types were found to accumulate within the peritubular capillaries and the interstitium, but amniotic fluid stem cells were more persistent than mesenchymal stem cells. In vitro experiments demonstrated that the two cell types produced different cytokines and growth factors, suggesting that a combination of different mediators is involved in their biological actions. These results suggest that the amniotic fluid-derived stem cells may improve renal regeneration in acute kidney injury, but they are not more effective than mesenchymal stem cells. PMID:20724594

Hauser, Peter V.; Fazio, Roberta De; Bruno, Stefania; Sdei, Simona; Grange, Cristina; Bussolati, Benedetta; Benedetto, Chiara; Camussi, Giovanni

2010-01-01

154

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

Microsoft Academic Search

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

K. Hara

1977-01-01

155

Abnormal glucose tolerance, white blood cell count, and telomere length in newly diagnosed, antidepressant-naïve patients with depression.  

PubMed

Chronic mood disorders have been associated with a shortened telomere, a marker of increased mortality rate and aging, 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 125.0mg/dL [67.9] vs 84.6 [25.6] (p<.001); for lymphocyte count 2.1×10(9)/L [0.6] vs 2.5×10(9)/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 aging disease. PMID:23207109

Garcia-Rizo, Clemente; Fernandez-Egea, Emilio; Miller, Brian J; Oliveira, Cristina; Justicia, Azucena; Griffith, Jeffrey K; Heaphy, Christopher M; Bernardo, Miguel; Kirkpatrick, Brian

2013-02-01

156

Abnormal Glucose Tolerance, White Blood Cell Count, and Telomere Length in Newly Diagnosed, Antidepressant-Na?ve Patients with Depression  

PubMed Central

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

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

157

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

NASA Astrophysics Data System (ADS)

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

Bel, J.; Marinoni, C.

2014-03-01

158

The relationship between antibiotic residue violations and somatic cell counts in Wisconsin dairy herds.  

PubMed

The objective of this retrospective observational study was to characterize somatic cell counts (SCC) on Wisconsin dairy farms and to determine the relationship between SCC and the risk of antibiotic residue violation. Monthly official state regulatory data were used when both the bulk tank SCC value and antibiotic test results were available for the same date. Data were collected from Wisconsin dairy farms from January 1995 through November 1998 and consisted of results of tests performed on 805,772 grade A and 176,763 grade B milk samples. Herd-year SCC averages were used to classify herds (< or =250,000; 251,000 to 400,000, 401,000 to 550,000, 551,000 to 700,000, >700,000), and the relative risk of antibiotic residue by SCC class was determined. Arithmetic mean SCC values were 334,634 and 480,029 for grade A and grade B milk, respectively. SCC values were significantly higher for samples with positive antibiotic residue tests for grade A milk during all 4 yr tested. The SCC values were significantly higher for samples with positive antibiotic residue tests for grade B milk for 3 of 4 yr. The rate of antibiotic residue violation per 1000 herd-years increased with SCC class for both grade A and grade B milk. The relative risks of antibiotic residue violation by SCC class were 1.0, 1.43, 2.38, 2.78, and 7.10 for grade A milk and 1.0, 1.11, 2.67, 4.33, and 5.43 for grade B milk. Programs to reduce the level of subclinical mastitis on dairy farms may have an additional benefit of reducing the risk of antibiotic residue violations. PMID:11132850

Ruegg, P L; Tabone, T J

2000-12-01

159

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

PubMed Central

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

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

2014-01-01

160

Amniotic Fluid Stem Cells from EGFP Transgenic Mice Attenuate Hyperoxia-Induced Acute Lung Injury  

PubMed Central

High concentrations of oxygen aggravate the severity of lung injury in patients requiring mechanical ventilation. Although mesenchymal stem cells have been shown to effectively attenuate various injured tissues, there is limited information regarding a role for amniotic fluid stem cells (AFSCs) in treating acute lung injury. We hypothesized that intravenous delivery of AFSCs would attenuate lung injury in an experimental model of hyperoxia-induced lung injury. AFSCs were isolated from EGFP transgenic mice. The in vitro differentiation, surface markers, and migration of the AFSCs were assessed by specific staining, flow cytometry, and a co-culture system, respectively. The in vivo therapeutic potential of AFSCs was evaluated in a model of acute hyperoxia-induced lung injury in mice. The administration of AFSCs significantly reduced the hyperoxia-induced pulmonary inflammation, as reflected by significant reductions in lung wet/dry ratio, neutrophil counts, and the level of apoptosis, as well as reducing the levels of inflammatory cytokine (IL-1?, IL-6, and TNF-?) and early-stage fibrosis in lung tissues. Moreover, EGFP-expressing AFSCs were detected and engrafted into a peripheral lung epithelial cell lineage by fluorescence microscopy and DAPI stain. Intravenous administration of AFSCs may offer a new therapeutic strategy for acute lung injury (ALI), for which efficient treatments are currently unavailable. PMID:24040409

Lai, Cheng-Wei; Yen, Chih-Ching; Lee, Kun-Hsiung; Wu, Shinn-Chih; Chen, Chuan-Mu

2013-01-01

161

Differentiation of stem\\/progenitor cells into vascular cells in response to fluid mechanical forces  

Microsoft Academic Search

Vascular functions are regulated not only by chemical mediators, such as hormones, cytokines, and neurotransmitters, but by\\u000a mechanical hemodynamic forces generated by blood flow and blood pressure. The mechanical force-mediated regulation is based\\u000a on the ability of vascular cells, including endothelial cells and smooth muscle cells, to recognize fluid mechanical forces,\\u000a i.e., the shear stress produced by flowing blood and

Kimiko Yamamoto; Joji Ando

2010-01-01

162

Human amniotic fluid stem cell differentiation along smooth muscle lineage.  

PubMed

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

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

2013-12-01

163

Nucleated red blood cells count as first prognostic marker for adverse neonatal outcome in severe preeclamptic pregnancies.  

PubMed

The purpose of this study was to determine acceptability of the nucleated red blood cells counts (NRBC) as early prognostic parameter for adverse outcome in preterm neonates born from pregnancies complicated with severe preeclampsia. We analysed 77 premature newborns who were born from pregnancies with severe preeclampsia during eight years (2004-2011) in our tertiary center. Women with other pregnancy complications were excluded from the study, as well as newborns with malformations and chromosomal anomalies. Newborns were compared according to the count of nucleated red blood cells (NRBC) on the first day of life. Cut off of NRBC was determined at 40 per 100 white blood cells. We analyzed and compared birth weight, gestational age, Apgar scores in 1st and 5th minute, hypoglycemia in first day of life, need for respiratory support, neonatal infection and brain ultrasound findings at the day of discharge between the groups of newborns. We found significantly lower birth weight, gestational age and Apgar scores in case group (NRB C > 40) and significantly higher rate of infections, need for respiratory support, abnormal brain ultrasound findings, morbidity rate and adverse neonatal outcome compared to control newborns group. Increased count of nucleated red blood cells (NRBC) in preterm newborns born from pregnancies with severe preeclampsia seems to be the first significant marker for detecting adverse neonatal outcome. PMID:23213944

Gasparovi?, Vesna Elvedi; Ahmetasevi?, Snjezana Gveri?; Coli?, Ana

2012-09-01

164

Patterns and predictors of CD4 T-cell counts among children born to HIV-infected women in Tanzania.  

PubMed

We assessed age-specific CD4 T-cell counts and their determinants among Tanzanian children born to HIV-infected mothers to address a major research gap. A total of 474 HIV-uninfected and 69 HIV-infected children were followed until age of 12 months. Maternal predictors were measured during pregnancy and child predictors at birth and throughout the follow up. Child CD4 T-cell counts were evaluated at the age of 3 months and subsequent 3-month intervals; they decreased linearly among HIV-infected (beta = -8 cells per week; 95% CI -12 to -4; P = 0.0003) and increased linearly among HIV-uninfected children (beta = 4 cells/week; 95% CI 2-7; P = 0.0008). Decreased child counts were predicted by low child anthropometry, maternal HIV stage > or =2, and maternal mid-upper arm circumference <27 cm among HIV-infected children; and by weight-for-height <-2 z-score, maternal HIV stage > or =2, maternal erythrocyte sedimentation rate <81 mm/h and maternal haemoglobin <8.5 g/dl among HIV-uninfected children. The maternal and child predictors described may serve as intervention targets among HIV-exposed children. PMID:19158163

Kupka, Roland; Msamanga, Gernard I; Aboud, Said; Manji, Karim P; Duggan, Christopher; Fawzi, Wafaie W

2009-10-01

165

Interleukin 10 Responses Are Associated With Sustained CD4 T-Cell Counts in Treated HIV Infection  

PubMed Central

Background.Inflammation persists in treated human immunodeficiency virus (HIV) infection and may contribute to an increased risk for non–AIDS-related pathologies. We investigated the correlation of cytokine responses with changes in CD4 T-cell levels and coinfection with hepatitis C virus (HCV) during highly active antiretroviral treatment (HAART). Methods.A total of 383 participants in the Women's Interagency HIV Study (212 with HIV monoinfection, 56 with HCV monoinfection, and 115 with HIV/HCV coinfection) were studied. HIV-infected women had <1000 HIV RNA copies/mL, 99.7% had >200 CD4 T cells/?L; 98% were receiving HAART at baseline. Changes in CD4 T-cell count between baseline and 2–4 years later were calculated. Peripheral blood mononuclear cells (PBMCs) obtained at baseline were used to measure interleukin 1? (IL-1?), interleukin 6 (IL-6), interleukin 10 (IL-10), interleukin 12 (IL-12), and tumor necrosis factor ? (TNF-?) responses to Toll-like receptor (TLR) 3 and TLR4 stimulation. Results.Undetectable HIV RNA (<80 copies/mL) at baseline and secretion of IL-10 by PBMCs were positively associated with gains in CD4 T-cell counts at follow-up. Inflammatory cytokines (IL-1?, IL-6, IL-12, and TNF-?) were also produced in TLR-stimulated cultures, but only IL-10 was significantly associated with sustained increases in CD4 T-cell levels. This association was significant only in women with HIV monoinfection, indicating that HCV coinfection is an important factor limiting gains in CD4 T-cell counts, possibly by contributing to unbalanced persistent inflammation. Conclusions.Secreted IL-10 from PBMCs may balance the inflammatory environment of HIV, resulting in CD4 T-cell stability. PMID:22693231

Villacres, Maria C.; Kono, Naoko; Mack, Wendy J.; Nowicki, Marek J.; Anastos, Kathryn; Augenbraun, Michael; Liu, Chenglong; Landay, Alan; Greenblatt, Ruth M.; Gange, Stephen J.; Levine, Alexandra M.

2012-01-01

166

Cardiac repair with injectable cell sheet fragments of human amniotic fluid stem cells in an immune-suppressed rat model  

Microsoft Academic Search

Direct intramyocardial injection of the desired cell types in a dissociated form is a common route of cell transplantation for repair of damaged myocardium. However, following injection of dissociated cells, a massive loss of transplanted cells has been reported. In this study, human amniotic fluid stem cells (hAFSCs) were used as the cell source for the fabrication of cell sheet

Yi-Chun Yeh; Wen-Yu Lee; Chu-Leng Yu; Shiaw-Min Hwang; Min-Fan Chung; Li-Wen Hsu; Yen Chang; Wei-Wen Lin; Ming-Song Tsai; Hao-Ji Wei; Hsing-Wen Sung

2010-01-01

167

Evaluation of the veterinary application of a point-of-care device measuring white blood cell counts.  

PubMed

A point-of-care device (POCD) for measuring total white blood cell count was evaluated for feline, canine, equine and bovine blood samples collected into EDTA. Mean biases were -9.2% (range, -12% to -6.3%) for feline samples, 20.2% (range, 15.3-25.1%) for canine samples, -7.1% (range, -8.3% to -5.9%) for equine samples, and 0.7% (range, -1.1% to 2.5%) for bovine samples. The results were influenced by the presence of nucleated red blood cells. The POCD provided precise, reliable data for feline, equine and bovine samples but the values obtained for the canine counts were overestimations. PMID:22503717

Riond, Barbara; Hofmann-Lehmann, Regina; Lutz, Hans

2012-10-01

168

Human Herpesvirus Replication and Abnormal CD8+ T Cell Activation and Low CD4+ T Cell Counts in Antiretroviral-Suppressed HIV-Infected Patients  

Microsoft Academic Search

BackgroundMost HIV-infected patients receiving virologically suppressive antiretroviral therapy continue to have abnormal, generalized T cell activation. We explored whether the degree of ongoing cytomegalovirus (CMV), Epstein-Barr virus (EBV) and Kaposi's sarcoma herpesvirus (KSHV) replication was associated with higher virus-specific T cell activation and the failure to achieve normal absolute CD4+ T cell counts in the face of long-term suppressive antiretroviral

Mark A. Jacobson; Dirk P. Ditmer; Elizabeth Sinclair; Jeffrey N. Martin; Steven G. Deeks; Peter Hunt; Edward S. Mocarski; Caroline Shiboski; Derya Unutmaz

2009-01-01

169

Correction of anaemia following renal transplantation: serial changes in serum immunoreactive erythropoietin, absolute reticulocyte count and red-cell creatine levels.  

PubMed

Improvement of erythropoiesis following successful human renal transplantation in eight patients was monitored by sequential measurements of haemoglobin, red-cell creatine, absolute reticulocyte count and estimates of serum immunoreactive erythropoietin (siEp). SiEp increased in five patients after transplant, in three cases almost immediately after a return to normal of plasma creatinine. The increase in siEp was followed by a rise in the absolute reticulocyte count and red-cell creatine and finally by an increase in the haemoglobin level. As the haemoglobin approached normal levels a decline in the absolute reticulocyte count preceded a fall in siEp levels. Red-cell creatine also fell, though more gradually than the reticulocyte count. Acute graft rejection (in two patients) was associated with a fall in siEp. Chronic rejection (in one patient) was associated with persistent increases in siEp, reticulocyte count and red-cell creatine; this patient subsequently developed erythrocytosis. PMID:3904812

Rejman, A S; Grimes, A J; Cotes, P M; Mansell, M A; Joekes, A M

1985-11-01

170

Blood cell counting in neonates: a comparison between a low volume micromethod and the standard laboratory method  

PubMed Central

Background Iatrogenic anaemia caused by repeated blood sampling to monitor laboratory parameters can contribute, particularly in neonates, to the need for transfusion. “Point of care” laboratory equipment uses smaller amounts of blood for analytic determinations and could, therefore, help to prevent secondary anaemia. In this study we compared the results of haematological parameters measured using a standard laboratory method and using a “point of care” micromethod, with the aim of validating the use of this latter method in clinical practice in neonatology. Materials and methods One hundred and fifty venous or capillary blood samples were taken from full-term or premature neonates 2–4 hours or 48 hours after birth. Each sample was processed by a standard haematology analyser and another micromethod instrument. Bland-Altman plots were constructed for each parameter and intra-class coefficients of correlation were calculated in order to evaluate the concordance between the two analysers. Results The concordance between the data obtained with the two analysers, expressed as the intra-class correlation, was 0.98 for white blood cell count, 0.97 for haemoglobin concentration, 0.96 for haematocrit, 0.95 for mean red cell volume and 0.98 for platelet count. The micromethod produced overestimated mean values for the leucocyte count (+1.27; p<0.001), haematocrit (+1.80; p<0.001) and platelet count (+13.55; p<0.001). Conclusions Overall, the concordance between the values obtained with the two analysers was high for each of the parameters taken into consideration. In the case of haemoglobin and leucocytes, give the high intra-class correlation and lack of systematic overestimation of one method over another, the micromethod guarantees a correct evaluation; however, despite the high intra-class correlations for platelet counts, the systemic error seems to suggest that the micromethod cannot guarantee an appropriate evaluation of this parameter. PMID:21839016

Papa, Fabrizio; Rongioletti, Mauro; Ventura, Marco Della; Di Turi, Francesco; Cortesi, Maurizio; Pasqualetti, Patrizio; Majolini, Maria Bernardetta; Collegiani, Valeria; Cicchese, Marika; Notarmuzi, Maria Letizia; Agostino, Rocco; Liumbruno, Giancarlo Maria

2011-01-01

171

Fluid mechanics, cell distribution, and environment in CellCube bioreactors.  

PubMed

Cultivation of MRC-5 cells and attenuated hepatitis A virus (HAV) for the production of VAQTA, an inactivated HAV vaccine (1), is performed in the CellCube reactor, a laminar flow fixed-bed bioreactor with an unusual diamond-shaped, diverging-converging flow geometry. These disposable bioreactors have found some popularity for the production of cells and gene therapy vectors at intermediate scales of operation (2, 3). Early testing of the CellCube revealed that the fluid mechanical environment played a significant role in nonuniform cell distribution patterns generated during the cell growth phase. Specifically, the reactor geometry and manufacturing artifacts, in combination with certain inoculum practices and circulation flow rates, can create cell growth behavior that is not simply explained. Via experimentation and computational fluid dynamics simulations we can account for practically all of the observed cell growth behavior, which appears to be due to a complex mixture of flow distribution, particle deposition under gravity, fluid shear, and possibly nutritional microenvironment. PMID:12572999

Auni?s, John G; Bader, Brett; Caola, Anthony; Griffiths, Janet; Katz, Maayan; Licari, Peter; Ram, Kripa; Ranucci, Colette S; Zhou, Weichang

2003-01-01

172

Risk factors for bulk milk somatic cell counts and total bacterial counts in smallholder dairy farms in the 10th region of Chile.  

PubMed

We investigated the principal management factors that influenced bulk milk somatic cell count (BMSCC) and total bacterial count (TBC) of smallholder dairy farms in the 10th region of Chile. One hundred and fifty smallholder milk producers were selected randomly from 42 milk collection centres (MCCs). In April and May of 2002, all farms were visited and a detailed interview questionnaire on dairy-cow management related to milk quality was conducted. In addition, the BMSCC and TBC results from the previous 2 months' fortnightly tests were obtained from the MCCs. The mean BMSCC and TBC were used as the dependent variables in the analyses and were normalised by a natural-logarithm transformation (LN). All independent management variables were categorised into binary outcomes and present (=1) was compared with absent (=0). Biserial correlations were calculated between the LNBMSCC or LNTBC and the management factors of the smallholder farms. Management factors with correlations with P0.05) factors. A random MCC effect was included in the models to investigate the importance of clustering of herds within MCC. In the null model for mean LNTBC, the random effect of MCCs was highly significant. It was explained by: milk collected once a day or less compared with collection twice a day, not cleaning the bucket after milking mastitic cows versus cleaning the bucket and cooling milk in a vat of water versus not cooling milk or using ice or a bulk tank to cool milk. Other factors that increased the LNTBC were a waiting yard with a soil or gravel floor versus concrete, use of plastic buckets for milking instead of metal, not feeding California mastitis test (CMT)-positive milk to calves and cows of dual-purpose breed. The final model explained 35% of the variance. The model predicted that a herd that complied with all the management practices had a mean predicted TBC of 105 colony forming units (cfu)/ml, whereas a herd that did not comply with any of these management factors had a predicted TBC of 59 x 10(9)cfu/ml. The model of mean LNBMSCC explained 18% of the variance; the random effect of MCC was not significant. Management factors that decreased the mean LNBMSCC were: using the CMT for 1 year versus using the test for more than 1 year or not at all, absence of a concrete waiting yard, not filtering the milk or using filters other than a plastic sieve to filter the milk, milking cows with mastitis last, and sometimes or always examining the udder before milking. A herd that complied with all of these management factors had a BMSCC of approximately 46,166 cells/ml, whereas a herd that did not comply with any of the management practices above had a mean BMSCC of 2 x 10(6)cells/ml. PMID:15698905

van Schaik, G; Green, L E; Guzmán, D; Esparza, H; Tadich, N

2005-01-01

173

Impact of Ambient Air Pollution on the Differential White Blood Cell Count in Patients with Chronic Pulmonary Disease  

PubMed Central

Epidemiologic studies report associations between particulate air pollution and increased mortality from pulmonary diseases.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 hours in association with all gaseous pollutants but showed no effect 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 hours before blood withdrawal and a second one with a time lag of 48-71 hours. 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. PMID:20064088

Bruske, Irene; Hampel, Regina; Socher, Martin M.; Ruckerl, Regina; Schneider, Alexandra; Heinrich, Joachim; Oberdorster, Gunter; Wichmann, H.-Erich; Peters, Annette

2013-01-01

174

Counting Books  

NSDL National Science Digital Library

The web site provides instructions for making counting books. Suggestions for using the completed books for counting one at a time, skip-counting, fractions and introducing addition and subtraction are given. Children should be able to write the numbers from 1 to 10 before beginning this activity.

2010-01-01

175

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

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

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

176

White Blood Cell Count in Women: Relation to Inflammatory Biomarkers, Haematological Profiles, Visceral Adiposity, and Other Cardiovascular Risk Factors  

PubMed Central

The role of white blood cell (WBC) count in pathogenesis of diabetes, cardiovascular disease, and obesity-related disorders has been reported earlier. Recent studies revealed that higher WBC contributes to atherosclerotic progression and impaired fasting glucose. However, it is unknown whether variations in WBC and haematologic profiles can occur in healthy obese individuals. The aim of this study is to further evaluate the influence of obesity on WBC count, inflammatory biomarkers, and metabolic risk factors in healthy women to establish a relationship among variables analyzed. The sample of the present study consisted of 84 healthy women with mean age of 35.56±6.83 years. They were categorized into two groups based on their body mass index (BMI): obese group with BMI >30 kg/m2 and non-obese group with BMI <30 kg/m2. We evaluated the relationship between WBC and platelet count (PLT) with serum interleukin 6 (IL-6), C-reactive protein (CRP), angiotensin ? (Ang ?), body fat percentage (BF %), waist-circumference (WC), and lipid profile. WBC, PLT, CRP, and IL-6 in obese subjects were significantly higher than in non-obese subjects (p< 0.05). The mean WBC count in obese subjects was 6.4±0.3 (×109/L) compared to 4.4±0.3 (×109/L) in non-obese subjects (p=0.035). WBC correlated with BF% (r=0.31, p=0.004), CRP (r=0.25, P=0.03), WC (r=0.22, p=0.04), angiotensin ? (r=0.24, p=0.03), triglyceride (r=0.24, p=0.03), and atherogenic index of plasma (AIP) levels (r=0.3, p=0.028) but not with IL-6. Platelet count was also associated with WC and waist-to-hip ratio (p<0.05). Haemoglobin and haematocrit were in consistent relationship with LDL-cholesterol (p<0.05). In conclusion, obesity was associated with higher WBC count and inflammatory parameters. There was also a positive relationship between WBC count and several inflammatory and metabolic risk factors in healthy women. PMID:23617205

Farhangi, Mahdieh Abbasalizad; Keshavarz, Seyyed-Ali; Eshraghian, Mohammadreza; Ostadrahimi, Alireza

2013-01-01

177

IVIg immune reconstitution treatment alleviates the state of persistent immune activation and suppressed CD4 T cell counts in CVID.  

PubMed

Common variable immunodeficiency (CVID) is characterized by defective B cell function, impaired antibody production, and increased susceptibility to bacterial infections. Here, we addressed the hypothesis that poor antibody-mediated immune control of infections may result in substantial perturbations in the T cell compartment. Newly diagnosed CVID patients were sampled before, and 6-12 months after, initiation of intravenous immunoglobulin (IVIg) therapy. Treatment-naïve CVID patients displayed suppressed CD4 T cell counts and myeloid dendritic cell (mDC) levels, as well as high levels of immune activation in CD8 T cells, CD4 T cells, and invariant natural killer T (iNKT) cells. Expression of co-stimulatory receptors CD80 and CD83 was elevated in mDCs and correlated with T cell activation. Levels of both FoxP3+ T regulatory (Treg) cells and iNKT cells were low, whereas soluble CD14 (sCD14), indicative of monocyte activation, was elevated. Importantly, immune reconstitution treatment with IVIg partially restored the CD4 T cell and mDC compartments. Treatment furthermore reduced the levels of CD8 T cell activation and mDC activation, whereas levels of Treg cells and iNKT cells remained low. Thus, primary deficiency in humoral immunity with impaired control of microbial infections is associated with significant pathological changes in cell-mediated immunity. Furthermore, therapeutic enhancement of humoral immunity with IVIg infusions alleviates several of these defects, indicating a relationship between poor antibody-mediated immune control of infections and the occurrence of abnormalities in the T cell and mDC compartments. These findings help our understanding of the immunopathogenesis of primary immunodeficiency, as well as acquired immunodeficiency caused by HIV-1 infection. PMID:24130688

Paquin-Proulx, Dominic; Santos, Bianca A N; Carvalho, Karina I; Toledo-Barros, Myrthes; Barreto de Oliveira, Ana Karolina; Kokron, Cristina M; Kalil, Jorge; Moll, Markus; Kallas, Esper G; Sandberg, Johan K

2013-01-01

178

Skip Counting With Counting Collections  

NSDL National Science Digital Library

In this 12-minute video Stephanie Latimer's kindergarten students develop strategies for counting collections of objects. They group objects, skip count, and record their results. The resource includes reflection questions for teacher discussion.

2012-01-01

179

Amniotic fluid cells are more efficiently reprogrammed to pluripotency than adult cells.  

PubMed

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

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; Polgar, Katalin

2010-04-01

180

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

PubMed Central

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

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

181

Self-Calibration of Cluster Dark Energy Studies: Counts in Cells  

E-print Network

Cluster number counts can constrain the properties of dark energy if and only if the evolution in the relationship between observable quantities and the cluster mass can be calibrated. Next generation surveys with ~10000 clusters will have sufficient statistics to enable some degree of self-calibration. The excess variance of counts due to the clustering of clusters provides such an opportunity and can be measured from the survey without additional observational cost. It can minimize the degradation in dark energy constraints due to an unknown power law evolution in the mass-observable relation improving constraints on the dark energy equation of state by a factor of 2 or more to sigma(w)=0.06 for a deep 4000 deg2 survey.

Marcos Lima; Wayne Hu

2004-01-26

182

Label-Free Differential Leukocyte Counts Using a Microfabricated, Single-Cell Impedance Spectrometer  

Microsoft Academic Search

Impedance spectroscopy is a non-invasive technique which allows analysis of the electrical properties of biological cells and other materials. Typically, the electrical properties of a cell suspension are measured using relatively large volumes (>100 mul). This approach suffers from the significant drawback that the measurement is averaged over several million cells, and as such, discrimination of cell sub-population and cell

David Holmes; Tao Sun; Hywel Morgan; Judith Holloway; Julie Cakebread; Donna Davies

2007-01-01

183

Cell separation and transportation between two miscible fluid streams using ultrasound  

PubMed Central

This paper presents a two-stream microfluidic system for transporting cells or micro-sized particles from one fluid stream to another by acoustophoresis. The two fluid streams, one being the original suspension and the other being the destination fluid, flow parallel to each other in a microchannel. Using a half-wave acoustic standing wave across the channel width, cells or particles with positive acoustic contrast factors are moved to the destination fluid where the pressure nodal line lies. By controlling the relative flow rate of the two fluid streams, the pressure nodal line can be maintained at a specific offset from the fluid interface within the destination fluid. Using this transportation method, particles or cells of different sizes and mechanical properties can be separated. The cells experiencing a larger acoustic radiation force are separated and transported from the original suspension to the destination fluid stream. The other particles or cells experiencing a smaller acoustic radiation force continue flowing in the original solution. Experiments were conducted to demonstrate the effective separation of polystyrene microbeads of different sizes (3??m and 10??m) and waterborne parasites (Giardia lamblia and Cryptosporidium parvum). Diffusion occurs between the two miscible fluids, but it was found to have little effects on the transport and separation process, even when the two fluids have different density and speed of sound. PMID:22662070

Liu, Yang; Hartono, Deny; Lim, Kian-Meng

2012-01-01

184

Highly Sensitive Automated Method for DNA Damage Assessment: Gamma-H2AX Foci Counting and Cell Cycle Sorting  

PubMed Central

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

Hernandez, Laia; Terradas, Mariona; Martin, Marta; Tusell, Laura; Genesca, Anna

2013-01-01

185

Inner ear stem cells derived feeder layer promote directional differentiation of amniotic fluid stem cells into functional neurons.  

PubMed

Intact spiral ganglion neurons are required for cochlear implantation or conventional hearing amplification as an intervention for sensorineural hearing loss. Treatment strategies to replace the loss of spiral ganglion neurons are needed. Recent reports have suggested that amniotic fluid-derived stem cells are capable of differentiating into neuron-like cells in response to cytokines and are not tumorigenic. Amniotic fluid stem cells represent a potential resource for cellular therapy of neural deafness due to spiral ganglion pathology. However, the directional differentiation of amniotic fluid stem cells is undetermined in the absence of cytokines and the consequence of inner ear supporting cells from the mouse cochlea organ of Corti on the differentiation of amniotic fluid stem cells remains to be defined. In an effort to circumvent these limitations, we investigated the effect of inner ear stem cells derived feeder layer on amniotic fluid stem cells differentiation in vitro. An inner ear stem cells derived feeder layer direct contact system was established to induce differentiation of amniotic fluid stem cells. Our results showed that inner ear stem cells derived feeder layer successfully promoted directional differentiation of amniotic fluid stem cells into neurons with characteristics of functionality. Furthermore, we showed that Wnt signaling may play an essential role in triggering neurogenesis. These findings indicate the potential use of inner ear stem cells derived feeder layer as a nerve-regenerative scaffold. A reliable and effective amniotic fluid stem cell differentiation support structure provided by inner ear stem cells derived feeder layer should contribute to efforts to translate cell-based strategies to the clinic. PMID:25124154

Zong, Ling; Chen, Kaitian; Zhou, Wei; Jiang, Di; Sun, Liang; Zhang, Xuemei; Jiang, Hongyan

2014-10-01

186

Pitted red cell counts in Nigerian children with sickle cell anemia: correlation with age and splenic size.  

PubMed Central

Using direct interference phase-contrast microscopy (Normansky Optics), pit counts were performed on 32 HbSS patients, aged 3 to 17 years. The influence of age and splenic size on counts were also investigated. Nine HbSS and 15 HbAA age and sex-matched, healthy individuals served as controls. The mean +/- SD counts in the three groups were 11.1 +/- 9.1%, 1.7 +/- 1.4% and 1.8 +/- 1.7%, respectively. The older SS patients tended to have higher values, but the linear correlation with age was not impressive (r = 0.28). Seventeen (53.1%) patients had counts greater than 10%, while 8 (25%) had less than 3.5%. Five patients with gross splenomegaly had a mean count of 4.3 +/- 1.9%, significantly lower than the figure of 12.3 +/- 7.9% for the patients without splenomegaly (P less than .001), demonstrating retained reticulo-endothelial function in such patients. PMID:1920505

Adekile, A. D.; Reindorf, C. A.; Adeodu, O. A.; Johnson, W.; Dairo, B. A.

1991-01-01

187

Satellited 4q identified in amniotic fluid cells  

SciTech Connect

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.

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

1995-01-16

188

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

NASA Astrophysics Data System (ADS)

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.

Yao, Wei; Ding, Guang-Hong

2011-08-01

189

Fluid phase endocytosis of [ 125 I]iodixanol in rat liver parenchymal, endothelial and Kupffer cells  

Microsoft Academic Search

Endocytosis of [125I]iodixanol was studied in vivo and in vitro in rat liver cells to determine fluid phase endocytic activity in different liver cells (hepatocytes, Kupffer cells and endothelial cells). The Kupffer cells were more active in the uptake of [125I]iodixanol than parenchymal cells or endothelial cells. Inhibition of endocytic uptake via clathrin-coated pits (by potassium depletion and hypertonic medium)

Rune Kjeken; Seyed Ali Mousavi; Andreas Brech; Tor Gjøen; Trond Berg

2001-01-01

190

Fluid Inclusion Gas Analysis  

SciTech Connect

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.

Lorie Dilley

2013-01-01

191

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

PubMed Central

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.

Motamedi, Ramin; Wood-Adams, Paula M.

2008-01-01

192

Counting Antigen-Specific CD8 T Cells: A Reevaluation of Bystander Activation during Viral Infection  

Microsoft Academic Search

Viral infections induce extensive T cell proliferation in vivo, but the specificity of the majority of the responding T cells has not been defined. To address this issue we used tetramers of MHC class I molecules containing viral peptides to directly visualize antigen-specific CD8 T cells during acute LCMV infection of mice. Based on tetramer binding and two sensitive assays

Kaja Murali-Krishna; John D. Altman; M. Suresh; David J. D. Sourdive; Allan J. Zajac; Joseph D. Miller; Jill Slansky; Rafi Ahmed

1998-01-01

193

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

Microsoft Academic Search

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

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

2007-01-01

194

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

E-print Network

The FASEB Journal · Research Communication Fluid shear stress primes mouse embryonic stem cells the increasing use of perfusion culture in stem cell research, it is unclear how changes in the stem cell stress is a ubiquitous environmen- tal cue experienced by stem cells when they are being differentiated

Voldman, Joel

195

Cryopreservation of amniotic fluid-derived stem cells using natural cryoprotectants and low concentrations of dimethylsulfoxide  

Microsoft Academic Search

Amniotic fluid-derived stem cells (AFSCs) are a potential cell source for therapeutic applications. They can be easily mass produced, cryopreserved and shipped to clinics for immediate use. However, one major obstacle to the manufacturing of clinical grade stem cells is the need for current good manufacturing practices for cryopreservation, storage, and distribution of these cells. Most current cryopreservation methods used

Ji Min Seo; Mi Yeung Sohn; Jang Soo Suh; Anthony Atala; James J. Yoo; Yun-Hee Shon

2011-01-01

196

Counting cell number in situ by quantification of dimethyl sulphide in culture headspace.  

PubMed

A novel, non-invasive technique is reported for determining the numbers of cells in a culture by quantifying dimethyl sulphide (DMS) in the culture headspace as produced by the cellular enzymatic reduction of dissolved dimethyl sulphoxide (DMSO). Measured DMS concentrations, as performed using selected ion flow tube mass spectrometry (SIFT-MS), in the headspace of 2D and 3D cultures of four cell lines, viz. HEK293 (kidney), MG63 (bone), hepG2 (liver) and CALU-1 (lung), linearly correlate with starting cell number. Clear differences in the rates of production of DMS by the four cell types in both the 2D and 3D situations are seen. This novel analytical technique for cell enumeration offers a significant contribution to quality assessment across cell-based research and industry, including analysis of large scale culture systems, and for routine cell biology research. PMID:25083513

Chippendale, Thomas W E; Span?l, Patrik; Smith, David; El Haj, Alicia J

2014-08-26

197

Isolation and Characterization of Porcine Amniotic Fluid-Derived Multipotent Stem Cells  

Microsoft Academic Search

The aim of this study was to isolate and characterize porcine amniotic fluid-derived multipotent stem cells (pAF-MSC). The porcine amniotic fluid (AF) from the amniotic cavity of pregnant gilts in the early stages of gestation (at E35) was collected and centrifuged for 5–10 min at 400 g to pellet cells. The primary culture of AF showed the multiple cell types,

Jiahuan Chen; Zhijuan Lu; De Cheng; Sha Peng; Huayan Wang

2011-01-01

198

Circulating tumor cell count during zoledronic acid treatment in men with metastatic prostate cancer: a pilot study  

PubMed Central

Purpose Recent clinical trials have demonstrated that zoledronic acid (ZOL) significantly prolongs survival in prostate cancer patients undergoing androgen deprivation therapy. This pilot study investigated the influence of ZOL on circulating tumor cell (CTC) counts in prostate cancer patients in association with prostate-specific antigen (PSA) used as a serum biomarker. Methods Patients with metastatic castration-resistant prostate cancer (CRPC) who were CTC-positive (n=4) were enrolled in treatment with ZOL between April 2012 and December 2013. CTCs were detected using the Cell Search System. The study evaluated CTC fluctuations at 1, 2, and 3 months versus baseline, as well as patient outcomes and adverse events. Results Two patients showed evidence of temporally decreased CTCs after ZOL treatment. Instead of decreasing the number of CTCs, the PSA level did not go down during the ZOL treatment. One patient could not undergo ZOL treatment due to rapid disease progression. Conclusions Although CTC count arguably provides useful information about patients undergoing ZOL treatment, the positive influence of ZOL may be limited to temporary effects for CRPC. PMID:25325027

Ide, Hisamitsu; Lu, Yan; Tanaka, Toshiaki; Wakumoto, Yoshiaki; Kitamura, Kosuke; Muto, Satoru; Yamaguchi, Raizo; Masumori, Naoya; Horie, Shigeo

2014-01-01

199

Single cell rheometry with a microfluidic constriction: quantitative control of friction and fluid leaks between cell and  

E-print Network

1 Single cell rheometry with a microfluidic constriction: quantitative control of friction and fluid leaks between cell and channel walls Pascal Preiraa , Marie-Pierre Valignata , José Bicob, France. We report how cell rheology measurements can be performed by monitoring the deformation of a cell

Paris-Sud XI, Université de

200

The Fluid-Kinetic Particle-in-Cell method for plasma simulations  

NASA Astrophysics Data System (ADS)

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

Markidis, Stefano; Henri, Pierre; Lapenta, Giovanni; Rönnmark, Kjell; Hamrin, Maria; Meliani, Zakaria; Laure, Erwin

2014-08-01

201

Simvastatin induces osteogenic differentiation in human amniotic fluid mesenchymal stem cells (AFMSC).  

PubMed

Amniotic fluid is a complex mixture composed of water, salts and different cells types derived from embryo exfoliation. Some of these cells present similar characteristics to mesenchymal stem cells as adherent properties, typical surface antigens and differentiation capacity. These cells are called amniotic fluid-derived mesenchymal stem cells (AFMSCs) and are easily obtained by amniocentesis, propagated in culture and differentiated in several cell types with specific inductions. In this study, we observe the ability of simvastatin, a 3-HMG-CoA reductase inhibitor, to induce AFSMCs osteogenic differentiation. When AFSMCs were incubated with medium containing simvastatin, it was observed morphological changes, calcium deposits formation confirmed by Alizarin Red stain. Differentiated cells also expressed typical osteogenic genes, as osteopontin and osteocalcin. In conclusion, simvastatin could be used as an optional osteogenic induction agent for amniotic fluid-derived mesenchymal stem cells. PMID:23094676

de Lara Janz, Felipe; Favero, Giovani Marino; Bohatch, Milton Sérgio; Aguiar Debes, Adrianade; Bydlowski, Sergio Paulo

2014-04-01

202

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

E-print Network

.................................................................................. 22 (a) Geodesic distance between two points in a sample region R (b) Geodesic influence zones ........................................................................... 31 23 (a) Topography at immersion level hmin (b) X at hmin (c) Topography... and compact in shape and neurons / glial cells that are larger, often hollow or exhibit a central dark spot inside a hollow ellipse, resembling an ‘eye’. There are three main types of cells in type B: granule cells that are tiny and compact, interneurons...

D'Souza, Aswin Cletus

2008-10-10

203

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

E-print Network

.................................................................................. 22 (a) Geodesic distance between two points in a sample region R (b) Geodesic influence zones ........................................................................... 31 23 (a) Topography at immersion level hmin (b) X at hmin (c) Topography... and compact in shape and neurons / glial cells that are larger, often hollow or exhibit a central dark spot inside a hollow ellipse, resembling an ?eye?. There are three main types of cells in type B: granule cells that are tiny and compact, interneurons...

D'Souza, Aswin Cletus

2009-05-15

204

CD4+ cell count recovery in na?ve patients initiating cART, who achieved and maintained plasma HIV-RNA suppression  

PubMed Central

Introduction A key objective of combined antiretroviral therapy (cART) is to reach and maintain high CD4 cell counts to provide long-term protection against AIDS-defining opportunistic infections and malignancies, as well as other comorbidities. However, a high proportion of patients present late for care. Our objective was to assess CD4 cell count recovery up to seven years in naïve patients initiating cART with at least three drugs in usual clinical care. Methods From the French Hospital Database on HIV, we selected naïve individuals initiating cART from 2000 with at least two years of follow-up. Participants were further required to have achieved viral load suppression by six months after initiating cART and were censored in case of virological failure. We calculated the proportion of patients (Kaplan-Meier estimates) who achieved CD4 recovery to >500/mm3 according to baseline CD4 cell count. Results A total of 15,025 patients were analyzed with a median follow-up on ART of 65.5 months (IQR: 42.3–96.0). At cART initiation, the median age was 38.6 years (IQR: 32.2–46.0), 9734 (64.8%) were men, median CD4 cell count was 239 (IQR: 130–336) and 2668 (17.8%) had a prior AIDS event. Results are presented in the Table 1. Conclusions This study shows that CD4 cell counts continue to increase seven years after cART initiation, whatever the baseline CD4 cell count. Failing to achieve CD4 recovery with continuous viral load suppression is rare for naïve patients initiating cART in routine clinical practice, but takes substantially longer in patients who initiate antiretroviral therapy at low CD4 cell counts. PMID:25393990

Costagliola, Dominique; Lacombe, Jean-Marc; Ghosn, Jade; Delaugerre, Constance; Pialoux, Gilles; Cuzin, Lise; Launay, Odile; Menard, Amelie; de Truchis, Pierre; Mary-Krause, Murielle; Weiss, Laurence; Delfraissy, Jean-Francois

2014-01-01

205

Counting Coins  

NSDL National Science Digital Library

We are learning about money and how to count coins. We need to learn about coins so we can pay for things we need to buy. These activities will help you practice counting money. Remember to record your learning as you work! Coin Paper We have been learning about coins. Listen to the coin song to remember the names of U.S. coins. U.S. Coin Song Before we can count coins, we need to know the names of the different coins and how much each coin is worth. Click the link below to review ...

Thorsen

2012-11-24

206

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

NASA Astrophysics Data System (ADS)

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.

Mitchell, Michael J.; King, Michael R.

2013-01-01

207

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

PubMed

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

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

2014-09-01

208

Microthermometric analysis of synthetic fluid inclusions in the hydrothermal diamond-anvil cell  

Microsoft Academic Search

The hydrothermal diamond-anvil cell (HDAC) was employed as a pressurized fluid inclusion heating stage to determine temperatures of phase transitions in synthetic fluid inclusions in quartz. Using this technique, the common problem of decrepitation or stretch- ing of inclusions having high internal pressures was eliminated. Homogenization temperatures of pure H2O synthetic inclusions determined in the HDAC are inversely related to

CHRISTIAN SCHMIDT; I-MING CHOU; ROBERT J. BODNAR; WILLIAM A. BASSETT

209

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

PubMed Central

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

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

2011-01-01

210

Counting carbohydrates  

MedlinePLUS

Carbohydrates are found in fruit, cereal, bread, pasta, and rice. They are quickly turned into a sugar ... sugar better if they can count how many carbohydrates they eat. Your dietitian will teach you a ...

211

Clock Counting  

NSDL National Science Digital Library

Students will practice telling time. Review clock counting with the interactive clock. Now match the clocks. Move over the hour clock to see if you chose correctly. Click the arrows to match the dragon clock to the written time. ...

Mcduffee, Ms.

2008-11-12

212

Flow manipulation and cell immobilization for biochemical applications using thermally responsive fluids  

PubMed Central

A flow redirection and single cell immobilization method in a microfluidic chip is presented. Microheaters generated localized heating and induced poly(N-isopropylacrylamide) phase transition, creating a hydrogel that blocked a channel or immobilized a single cell. The heaters were activated in sets to redirect flow and exchange the fluid in which an immobilized cell was immersed. A yeast cell was immobilized in hydrogel and a 4?,6-diamidino-2-phenylindole (DAPI) fluorescent stain was introduced using flow redirection. DAPI diffused through the hydrogel and fluorescently labelled the yeast DNA, demonstrating in situ single cell biochemistry by means of immobilization and fluid exchange. PMID:24285990

Haraksingh Thilsted, Anil; Bazargan, Vahid; Piggott, Nina; Measday, Vivien; Stoeber, Boris

2012-01-01

213

Flow manipulation and cell immobilization for biochemical applications using thermally responsive fluids.  

PubMed

A flow redirection and single cell immobilization method in a microfluidic chip is presented. Microheaters generated localized heating and induced poly(N-isopropylacrylamide) phase transition, creating a hydrogel that blocked a channel or immobilized a single cell. The heaters were activated in sets to redirect flow and exchange the fluid in which an immobilized cell was immersed. A yeast cell was immobilized in hydrogel and a 4',6-diamidino-2-phenylindole (DAPI) fluorescent stain was introduced using flow redirection. DAPI diffused through the hydrogel and fluorescently labelled the yeast DNA, demonstrating in situ single cell biochemistry by means of immobilization and fluid exchange. PMID:24285990

Haraksingh Thilsted, Anil; Bazargan, Vahid; Piggott, Nina; Measday, Vivien; Stoeber, Boris

2012-01-01

214

Antral G- and D-cell counts in chronic renal failure.  

PubMed

Antral somatostatin- and gastrin-producing cells (D and G cells) were studied in a group of patients with chronic renal failure (CRF) in comparison with a control group. Gastric acid secretion and serum gastrin, phosphate, and parathormone (PTH) levels were also evaluated in every patient. The group with CRF showed a mild increase both in G- and in D-cell denisty. In this group serum phosphate and PTH levels were higher than normal, showing hyperparathyroidism in every patient. A direct correlation was found between G-cell density and parathyroid function in patients with CRF. Hyperparathyroidism, therefore, seems to play a role in the mechanism of increased serum gastrin levels in CRF. PMID:375375

Crivelli, O; Pera, A; Lombardo, L; Vernero, S; Varetto, H; Fruttero, B; Giorcelli, G; Babando, G; Verme, G

1979-01-01

215

Single-molecule transcript counting of stem-cell markers in the mouse intestine  

E-print Network

Determining the molecular identities of adult stem cells requires technologies for sensitive transcript detection in tissues. In mouse intestinal crypts, lineage-tracing studies indicated that different genes uniquely mark ...

Itzkovitz, Shaul Shalev

216

Choral Counting  

NSDL National Science Digital Library

As a whole group, have students chant the counting sequence starting with one to thirty, using the pointer to follow the number sequence. Over time, increase the range to one to fifty and then one to one hundred. Eventually have a student take over the job of pointing out the numbers in the sequence. Highlight the multiples of ten using a marker or a colored screen and have students chant the counting sequence by 10s. This should be done daily.

Mathematics, Illustrative

2012-07-31

217

Home-based versus clinic-based care for patients starting antiretroviral therapy with low CD4+ cell counts: findings from a cluster-randomized trial  

PubMed Central

Objectives: African health services have shortages of clinical staff. We showed previously, in a cluster-randomized trial, that a home-based strategy using trained lay-workers is as effective as a clinic-based strategy. It is not known whether home-based care is suitable for patients with advanced HIV disease. Methods: The trial was conducted in Jinja, Uganda. One thousand, four hundred and fifty-three adults initiating ART between February 2005 and January 2009 were randomized to receive either home-based care or routine clinic-based care, and followed up for about 3 years. Trained lay workers, supervised by clinical staff based in a clinic, delivered the home-based care. In this sub-analysis, we compared survival between the two strategies for those who presented with CD4+ cell count less than 50 cells/?l and those who presented with higher CD4+ cell counts. We used Kaplan–Meier methods and Poisson regression. Results: Four hundred and forty four of 1453 (31%) participants had baseline CD4+ cell count less than 50?cells/?l. Overall, 110 (25%) deaths occurred among participants with baseline CD4+ cell count less than 50?cells/?l and 87 (9%) in those with higher CD4+ cell count. Among participants with CD4+ cell count less than 50?cells/?l, mortality rates were similar for the home and facility-based arms; adjusted mortality rate ratio 0.80 [95% confidence interval (CI) 0.53–1.18] compared with 1.22 (95% CI 0.78–1.89) for those who presented with higher CD4+ cell count. Conclusion: HIV home-based care, with lay workers playing a major role in the delivery of care including providing monthly adherence support, leads to similar survival rates as clinic-based care even among patients who present with very low CD4+ cell count. This emphasises the critical role of adherence to antiretroviral therapy. PMID:24468997

Woodd, Susannah L.; Grosskurth, Heiner; Levin, Jonathan; Amuron, Barbara; Namara, Geoffrey; Birunghi, Josephine; Coutinho, Alex; Jaffar, Shabbar

2014-01-01

218

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

PubMed Central

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

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

2014-01-01

219

Flow cytometry total cell counts: a field study assessing microbiological water quality and growth in unchlorinated drinking water distribution systems.  

PubMed

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 (R (2) = 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

Liu, G; Van der Mark, E J; Verberk, J Q J C; Van Dijk, J C

2013-01-01

220

BANA-Positive Plaque Samples Are Associated with Oral Hygiene Practices and Not CD4+ T Cell Counts in HIV-Positive Patients  

PubMed Central

Background. The “red complex” microorganisms, namely, Porphyromonas gingivalis, Treponema denticola, and Tannerella forsythia are considered as potential pathogens causing HIV-associated periodontal diseases. Moreover, it has been recognized that an association exists between CD4+ T cell counts and periodontal disease progression. Objective. To establish whether CD4+ T cell counts or oral hygiene plays a greater role in producing BANA-positive results in HIV-associated periodontal disease. Materials and Methods. One hundred and twenty HIV-positive patients participated in the study, and their CD4+ T cell counts were obtained from their medical records. The six Ramfjord teeth were used for evaluating periodontal clinical indices and subgingival plaque sampling. BANA test was used for the detection and prevalence of the “red complex” bacteria in plaque samples. Results. A majority of 69.17% HIV-positive patients were BANA-positive. No significant associations were found between BANA and CD4+ T cell counts. A highly significant association was found between BANA with probing depth and clinical attachment level (P ? 0.0001) and between BANA and the use of interdental aids (P = 0.0168). Conclusion. HIV-associated periodontal diseases are strongly related to oral hygiene practices rather than the effect of CD4+ T cell counts, and the use of interdental aids was marked as a significant predictor of BANA-negative plaque samples. PMID:23258979

John, Cathy Nisha; Xavier Graham Stephen, Lawrence; Wilma Joyce Africa, Charlene

2012-01-01

221

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

E-print Network

and regeneration. We find that the expression of all markers overlaps at crypt-base cells. This co of the markers following irradiation, indicating their potential role in the regeneration process. Our approachT E C H N I C A L R E P O R T Single-molecule transcript counting of stem-cell markers in the mouse

van Oudenaarden, Alexander

222

Angiotensin Peptide Regulation of Fluid-Phase Endocytosis in Brain Microvessel Endothelial Cell Monolayers  

Microsoft Academic Search

Summary: An in vitro model comprised of primary cultures of brain microvessel endothelial cells was used to investigate angiotensin II (Ang II) effects on blood-brain barrier fluid-phase endocytosis. The effects of Ang II, saralasin, sarathrin, bradykinin (BK), and phorbol myristate acetate (PMA) on brain microvessel endothelial cell fluid-phase endocytosis were determined using the fluorescent marker, Lucifer yellow. Nanomolar concentrations of

François L. Guillot; Kenneth L. Audus

1990-01-01

223

Deferoxamine compensates for decreases in B cell counts and reduces mortality in enterovirus 71-infected mice.  

PubMed

Enterovirus 71 is one of the major causative agents of hand, foot and mouth disease in children under six years of age. No vaccine or antiviral therapy is currently available. In this work, we found that the number of B cells was reduced in enterovirus 71-infected mice. Deferoxamine, a marine microbial natural product, compensated for the decreased levels of B cells caused by enterovirus 71 infection. The neutralizing antibody titer was also improved after deferoxamine treatment. Furthermore, deferoxamine relieved symptoms and reduced mortality and muscle damage caused by enterovirus 71 infection. This work suggested that deferoxamine has the potential for further development as a B cell-immunomodulator against enterovirus 71. PMID:25003792

Yang, Yajun; Ma, Jing; Xiu, Jinghui; Bai, Lin; Guan, Feifei; Zhang, Li; Liu, Jiangning; Zhang, Lianfeng

2014-07-01

224

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

PubMed

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. PMID:22480746

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

2012-10-01

225

Amniotic membrane and amniotic fluid-derived cells: potential tools for regenerative medicine?  

PubMed

Human amniotic membranes and amniotic fluid have attracted increasing attention in recent years as a possible reserve of stem cells that may be useful for clinical application in regenerative medicine. Many studies have been conducted to date in terms of the differentiation potential of these cells, with several reports demonstrating that cells from both the amniotic fluid and membrane display high plasticity. In addition, cells from the amniotic membrane have also been shown to display immunomodulatory characteristics both in vivo and in vitro, which could make them useful in an allotransplantation setting. Here, we provide an overview comparing the latest findings regarding the stem characteristics of cells from both the amniotic membrane and amniotic fluid, as well as on the potential utility of these cells for future clinical application in regenerative medicine. PMID:19317646

Parolini, Ornella; Soncini, Maddalena; Evangelista, Marco; Schmidt, Dörthe

2009-03-01

226

Effect of cooling rate on eutectic cell count, grain size, microstructure, and ultimate tensile strength of hypoeutectic cast iron  

NASA Astrophysics Data System (ADS)

This article describes a series of microstructural and strength studies performed on hypoeutectic cast iron, which was sand cast using a variety of end chills (metallic, nonmetallic, water-cooled, and subzero, respectively). The effects of cooling rate on the eutectic cell count (ECC), grain size, and the ultimate tensile strength (UTS) were evaluated. Attempts were also made to explain these effects and to correlate the UTS with ECC. It was found that subzero chilled and water-cool, chilled cast iron exhibit severe undercooling compared to normal sand cast iron. It was concluded from this investigation that nucleation conditions are completely altered but growth conditions prevail as usual. Therefore, undercooling during solidification is considered to be responsible for variation in ECC, grain size, microstructure, and tensile strength.

Hemanth, J.; Rao, K. V. S.

1999-08-01

227

Induction of E-cadherin+ human amniotic fluid cell differentiation into oocyte-like cells via culture in medium supplemented with follicular fluid.  

PubMed

Pluripotent human amniotic fluid cells (HuAFCs) can differentiate into various types of somatic cell in vitro. However, their differentiation into oocyte-like cells has never been described to the best of our knowledge. In the present study, differentiation of E-cadherin+ and E-cadherin- HuAFC sub-populations into oocyte-like cells was induced via culture in medium containing bovine follicular fluid and ?-mercaptoethanol. The E-cadherin+ HuAFCs expressed DAZL highly. Post-induction, cells with an oocyte-like phenotype were found among the E-cadherin+ HuAFCs, expressing markers specific to germ cells and oocytes (VASA, ZP3 and GDF9) and meiosis (DMC1 and SCP3). When specific small interfering RNA (siRNA) was used to suppress E-cadherin in the E-cadherin+ HuAFCs, the levels of DAZL expression were reduced. Post-induction, the morphology of the siRNA?E?cadherin HuAFCs was poorer and the expression levels of germ cell-specific markers were lower compared with those of the siRNA-mock HuAFCs. Therefore, E-cadherin+ HuAFCs could be more easily induced to differentiate into oocyte-like cells by bovine follicular fluid and ?-mercaptoethanol. In addition, the E-cadherin+ HuAFCs exhibited potential characteristics of DAZL protein expression, and thus it was conjectured that bovine follicular fluid acts on DAZL protein and promotes E-cadherin+ HuAFC differentiation into oocyte-like cells. PMID:24788191

Liu, Te; Huang, Yongyi; Bu, Yanzhen; Zhao, Yanhui; Zou, Gang; Liu, Zhixue

2014-07-01

228

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

PubMed

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. PMID:24852600

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

2014-05-01

229

Elevated White Blood Cell Count Is Associated with Higher Risk of Glucose Metabolism Disorders in Middle-Aged and Elderly Chinese People  

PubMed Central

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. PMID:24852600

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

2014-01-01

230

AIDS . Author manuscript HIV-specific regulatory T cells are associated with higher CD4 cell counts  

E-print Network

to chronic patients (n 19). They+ low high = exhibited a phenotype of memory T cells and expressed ; methods ; Cell Proliferation ; Female ; Flow Cytometry ; HIV Infections ; diagnosis ; immunology ; immunology ; Male ; Phenotype ; Prospective Studies ; RNA, Viral ; immunology ; metabolism ; Suppressor

Paris-Sud XI, Université de

231

Apparatus for filling the cavities of cells and laminated substrates with a fluid  

DOEpatents

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.

Lopez Tonazzi, Juan C. (Tucson, AZ); Kucharczyk, Jr., Joseph E. (Tucson, AZ); Agrawal, Anoop (Tucson, AZ)

2001-01-01

232

Method for filling the cavities of cells with a chromogenic fluid  

DOEpatents

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.

Tonazzi, Juan C. Lopez (Tucson, AZ); Kucharczyk, Jr., Joseph E. (Tucson, AZ); Agrawal, Anoop (Tucson, AZ)

1999-01-01

233

Method for filling the cavities of cells with a chromogenic fluid  

DOEpatents

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.

Tonazzi, J.C.L.; Kucharczyk, J.E. Jr.; Agrawal, A.

1999-01-05

234

Sterile Fluid Collections in Acute Pancreatitis: Catheter Drainage Versus Simple Aspiration  

Microsoft Academic Search

Purpose. To compare the clinical outcome of needle aspiration versus percutaneous catheter drainage of sterile fluid collections in patients with acute pancreatitis. Methods. We reviewed the clinical and imaging data of patients with acute pancreatic fluid collections from 1998 to 2003. Referral for fluid sampling was based on elevated white blood cell count and fevers. Those patients with culture-negative drainages

Eric M. Walser; William H. Nealon; Santiago Marroquin; Syed Raza; J. Alberto Hernandez; James Vasek

2006-01-01

235

Reduction in Early Mortality on Antiretroviral Therapy for Adults in Rural South Africa Since Change in CD4+ Cell Count Eligibility Criteria  

PubMed Central

Objective: To explore the impact of expanded eligibility criteria for antiretroviral therapy (ART) on median CD4+ cell count at ART initiation and early mortality on ART. Methods: Analyses included all adults (?16 years) initiated on first-line ART between August 2004 and July 2012. CD4+ cell count threshold 350 cells per microliter for all adults was implemented in August 2011. Early mortality was defined as any death within 91 days of ART initiation. Trends in baseline CD4+ cell count and early mortality were examined by year (August to July) of ART initiation. Competing risks analysis was used to examine early mortality. Results: A total of 19,080 adults (67.6% female) initiated ART. Median CD4+ cell count at ART initiation was 110–120 cells per microliter over the first 6 years, increasing marginally to 145 cells per microliter in 2010–2011 and more significantly to 199 cells per microliter in 2011–2012. Overall, there were 875 deaths within 91 days of ART initiation; early mortality rate was 19.4 per 100 person-years [95% confidence interval (CI) 18.2 to 20.7]. After adjustment for sex, age, baseline CD4+ cell count, and concurrent tuberculosis (TB), there was a 46% decrease in early mortality for those who initiated ART in 2011–2012 compared with the reference period 2008–2009 (subhazard ratio, 0.54; 95% CI: 0.41 to 0.71). Conclusions: Since the expansion of eligibility criteria, there is evidence of earlier access to ART and a significant reduction in early mortality rate in this primary health care programme. These findings provide strong support for national ART policies and highlight the importance of earlier ART initiation for achieving reductions in HIV-related mortality. PMID:23756374

Mutevedzi, Portia C.; Iwuji, Collins C.; Newell, Marie-Louise

2014-01-01

236

Mesenchymal cells from human amniotic fluid survive and migrate after transplantation into adult rat brain.  

PubMed

Amniotic fluid has been recently suggested as an alternative source of mesenchymal stem cells. However, the fate of amniotic fluid-derived mesenchymal stem cells (AF-MSCs) after in vivo transplantation has yet to be determined. In the present study we explored whether human AF-MSCs could survive and migrate following transplantation into the striatum of normal and ischemic rat. We found that the grafted cells could survive and migrate towards multiple brain regions in the normal animals, while they moved towards the injured region in the ischemic rat. Double-immunostaining analyses showed that the implanted human AF-MSCs express markers for immature neurons (Doublecortin) at 10 days, and for astrocytes (GFAP) at 10, 30 and 90 after transplantation. This study provides the first evidence that human amniotic fluid contains cells having the potential to survive and integrate into adult rat brain tissue and, therefore, to function as effective stem cells for therapeutic strategies. PMID:17379545

Cipriani, Sabrina; Bonini, Daniela; Marchina, Eleonora; Balgkouranidou, Ioanna; Caimi, Luigi; Grassi Zucconi, Gigliola; Barlati, Sergio

2007-08-01

237

Human transgene-free amniotic-fluid-derived induced pluripotent stem cells for autologous cell therapy.  

PubMed

The establishment of a reliable prenatal source of autologous, transgene-free progenitor cells has enormous potential in the development of regenerative-medicine-based therapies for infants born with devastating birth defects. Here, we show that a largely CD117-negative population of human amniotic fluid mesenchymal stromal cells (AF-MSCs) obtained from fetuses with or without prenatally diagnosed anomalies are readily abundant and have limited baseline differentiation potential when compared with bone-marrow-derived MSCs and other somatic cell types. Nonetheless, the AF-MSCs could be easily reprogrammed into induced pluripotent stem cells (iPSCs) using nonintegrating Sendai viral vectors encoding for OCT4, SOX2, KLF4, and cMYC. The iPSCs were virtually indistinguishable from human embryonic stem cells in multiple assays and could be used to generate a relatively homogeneous population of neural progenitors, expressing PAX6, SOX2, SOX3, Musashi-1, and PSA-NCAM, for potential use in neurologic diseases. Further, these neural progenitors showed engraftment potential in vivo and were capable of differentiating into mature neurons and astrocytes in vitro. This study demonstrates the usefulness of AF-MSCs as an excellent source for the generation of human transgene-free iPSCs ideally suited for autologous perinatal regenerative medicine applications. PMID:25014361

Jiang, Guihua; Di Bernardo, Julie; Maiden, Michael M; Villa-Diaz, Luis G; Mabrouk, Omar S; Krebsbach, Paul H; O'Shea, K Sue; Kunisaki, Shaun M

2014-11-01

238

CD4 cell count recovery among HIV-infected patients with very advanced immunodeficiency commencing antiretroviral treatment in sub-Saharan Africa  

Microsoft Academic Search

BACKGROUND: Patients accessing antiretroviral treatment (ART) programmes in sub-Saharan Africa frequently have very advanced immunodeficiency. Previous data suggest that such patients may have diminished capacity for CD4 cell count recovery. METHODS: Rates of CD4 cell increase were determined over 48 weeks among ART-naïve individuals (n = 596) commencing ART in a South African community-based ART programme. RESULTS: The CD4 cell

Stephen D Lawn; Landon Myer; Linda-Gail Bekker; Robin Wood

2006-01-01

239

Optical Detection and Virotherapy of Live Metastatic Tumor Cells in Body Fluids with Vaccinia Strains  

PubMed Central

Metastatic tumor cells in body fluids are important targets for treatment, and critical surrogate markers for evaluating cancer prognosis and therapeutic response. Here we report, for the first time, that live metastatic tumor cells in blood samples from mice bearing human tumor xenografts and in blood and cerebrospinal fluid samples from patients with cancer were successfully detected using a tumor cell-specific recombinant vaccinia virus (VACV). In contrast to the FDA-approved CellSearch system, VACV detects circulating tumor cells (CTCs) in a cancer biomarker-independent manner, thus, free of any bias related to the use of antibodies, and can be potentially a universal system for detection of live CTCs of any tumor type, not limited to CTCs of epithelial origin. Furthermore, we demonstrate for the first time that VACV was effective in preventing and reducing circulating tumor cells in mice bearing human tumor xenografts. Importantly, a single intra-peritoneal delivery of VACV resulted in a dramatic decline in the number of tumor cells in the ascitic fluid from a patient with gastric cancer. Taken together, these results suggest VACV to be a useful tool for quantitative detection of live tumor cells in liquid biopsies as well as a potentially effective treatment for reducing or eliminating live tumor cells in body fluids of patients with metastatic disease. PMID:24019862

Minev, Boris R.; Zimmermann, Martina; Aguilar, Richard J.; Zhang, Qian; Sturm, Julia B.; Fend, Falko; Yu, Yong A.; Cappello, Joseph; Lauer, Ulrich M.; Szalay, Aladar A.

2013-01-01

240

Counting Circles  

NSDL National Science Digital Library

Have students stand and form a circle facing in toward each other. Select a counting sequence to be practiced with no more than 8-10 numbers in the sequence. Have the students start counting around the circle one by one until the last number in the sequence is reached. When the last number is reached all students clap and that student is out and sits down on the floor in the middle of the circle. Start the counting sequence over again until another student reaches the number at the end of the sequence; everyone claps and that student sits in the center with the first student. Continue repeating the sequence until only one child is left standing and the rest are seated in the center of the circle. For example: for the counting sequence 1-10: the first student says "one," the next student says "two" and so on until the 10th students gets to "ten" at this point everyone claps and the tenth child sits in the center of the circle. The eleventh student starts over with "one" and so on.

Mathematics, Illustrative

2012-07-31

241

Counting Coins  

NSDL National Science Digital Library

In this iOS app students practice counting U.S. coins by matching the value, making the total, telling how much, and creating their own values. Students drag coins onto a digital mat or enter values with a keypad to complete the tasks, and then receive feedback.

K12, Inc.

2011-03-23

242

Genetic variants associated with the white blood cell count in 13,923 subjects in the eMERGE Network  

PubMed Central

White blood cell count (WBC) is unique among identified inflammatory predictors of chronic disease in that it is routinely measured in asymptomatic patients in the course of routine patient care. We led a genome-wide association analysis to identify variants associated with WBC levels in 13,923 subjects in the electronic Medical Records and Genomics (eMERGE) Network. We identified two regions of interest that were each unique to subjects of genetically determined ancestry to the African continent (AA) or to the European continent (EA). WBC varies among different ancestry groups. Despite being ancestry specific, these regions were identifiable in the combined analysis. In AA subjects, the region surrounding the Duffy antigen/chemokine receptor gene (DARC) on 1q21 exhibited significant association (p value = 6.71e–55). These results validate the previously reported association between WBC and of the regulatory variant rs2814778 in the promoter region, which causes the Duffy negative phenotype (Fy?/?). A second missense variant (rs12075) is responsible for the two principal antigens, Fya and Fyb of the Duffy blood group system. The two variants, consisting of four alleles, act in concert to produce five antigens and subsequent phenotypes. We were able to identify the marginal and novel interaction effects of these two variants on WBC. In the EA subjects, we identified significantly associated SNPs tagging three separate genes in the 17q21 region: (1) GSDMA, (2) MED24, and (3) PSMD3. Variants in this region have been reported to be associated with WBC, neutrophil count, and inflammatory diseases including asthma and Crohn’s disease. PMID:22037903

McDavid, Andrew; Weston, Noah; Nelson, Sarah C.; Zheng, Xiuwen; Hart, Eugene; de Andrade, Mariza; Kullo, Iftikhar J.; McCarty, Catherine A.; Doheny, Kimberly F.; Pugh, Elizabeth; Kho, Abel; Hayes, M. Geoffrey; Pretel, Stephanie; Saip, Alexander; Ritchie, Marylyn D.; Crawford, Dana C.; Crane, Paul K.; Newton, Katherine; Li, Rongling; Mirel, Daniel B.; Crenshaw, Andrew; Larson, Eric B.; Carlson, Chris S.; Jarvik, Gail P.

2013-01-01

243

Stability of blood cell counts, hematologic parameters and reticulocytes indexes on the Advia A120 hematologic analyzer.  

PubMed

Delayed sample analysis is not a rare circumstance in clinical and laboratory practice, especially when blood samples are shipped to distant centralized laboratories, when the analysis can not be readily performed, or when retesting is appropriate. In this study we sought to evaluate the stability of conventional and new hematologic parameters in blood specimens stored for as long as 24 hours at 4 degrees C. Of the 21 hematologic parameters tested with the use of the Advia 120 hematologic analyzer (Bayer Diagnostics), means for paired samples of specimens differed significantly over the 24-hour storage period for hematocrit, main corpuscular volume, percentage of macrocytes, platelet count, main platelet volume, reticulocyte count and percentage, and reticulocyte hemoglobin content (all P < .01). We noted no significant changes in the other parameters tested or in the white blood cell differential. The overall distribution of the immature reticulocytes fractions remained substantially unchanged, though the high staining-intensity fraction showed a considerable shift from the baseline measure. Bland-Altman plots and limits-of-agreement analysis showed mean biases between -4.8% and 37.2% and relative coefficients of variations ranging from 0.4% to 32.7%. The 95% agreement interval in the set of differences was satisfactory and almost within the current analytic-quality specifications for desirable bias. The results of this investigation suggest that, within certain limitations for parameters derived or calculated from cellular volumes, blood specimens stored for as long as 24 hours at 4 degrees C may be suitable for hematologic testing. PMID:16310516

Lippi, Giuseppe; Salvagno, Gian Luca; Solero, Gian Pietro; Franchini, Massimo; Guidi, Gian Cesare

2005-12-01

244

Emergent long-range couplings in arrays of fluid cells.  

PubMed

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 confirmed by Monte Carlo 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. PMID:25170731

Abraham, D B; Macio?ek, A; Vasilyev, O

2014-08-15

245

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

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

Hews, Diana K.; Hara, Erina; Anderson, Maurice C.

2012-01-01

246

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

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/?l 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 differentially influences periodontal disease both before and after HAART in HIV-infected adults. PMID:24146949

Vernon, Lance T.; Demko, Catherine A.; Babineau, Denise C.; Wang, Xuelei; Toossi, Zahra; Weinberg, Aaron; Rodriguez, Benigno

2013-01-01

247

Resistance to Fluid Shear Stress Is a Conserved Biophysical Property of Malignant Cells  

PubMed Central

During metastasis, cancer cells enter the circulation in order to gain access to distant tissues, but how this fluid microenvironment influences cancer cell biology is poorly understood. A longstanding view is that circulating cancer cells derived from solid tissues may be susceptible to damage from hemodynamic shear forces, contributing to metastatic inefficiency. Here we report that compared to non-transformed epithelial cells, transformed cells are remarkably resistant to fluid shear stress (FSS) in a microfluidic protocol, exhibiting a biphasic decrease in viability when subjected to a series of millisecond pulses of high FSS. We show that magnitude of FSS resistance is influenced by several oncogenes, is an adaptive and transient response triggered by plasma membrane damage and requires extracellular calcium and actin cytoskeletal dynamics. This novel property of malignant cancer cells may facilitate hematogenous metastasis and indicates, contrary to expectations, that cancer cells are quite resistant to destruction by hemodynamic shear forces. PMID:23226552

Henry, Michael D.

2012-01-01

248

Handling solid-fluid interfaces for viscous flows: explicit jump approximation vs. ghost cell approaches  

E-print Network

Handling solid-fluid interfaces for viscous flows: explicit jump approximation vs. ghost cell University, Jhongli, Taiwan Abstract The ghost cell approaches (GCA) for handling stationary solid boundaries- trapolation for enforcing no-slip boundary conditions should be avoided, even for regular solid boundaries

249

Regulation of bronchoalveolar lavage fluids cell function by the immunomodulatory agents from Cordyceps sinensis  

Microsoft Academic Search

Cordyceps sinensis (C. sinensis) is one of the well known fungi used in traditional Chinese medicine for treatment asthma and bronchial and lung inflammation. In this study, effects of C. sinensis methanolic extracts on bronchoalveolar lavage fluids (BALF) cells proliferation, inflammatory cytokines production, and genes expression were evaluated. The proliferative response of BALF cells to lipopolysaccharide (LPS) was determined by

Yuh-Chi Kuo; Wei-Jern Tsai; Jir-Yenn Wang; Shi-Chung Chang; Ching-Yuang Lin; Ming-Shi Shiao

2001-01-01

250

Photoacoustic and photothermal detection of circulating tumor cells, bacteria and nanoparticles in cerebrospinal fluid in vivo and ex vivo  

PubMed Central

Circulating cells, proteins, microparticles, and DNA in cerebrospinal fluid (CSF) are excellent biomarkers of many diseases, including cancer and infections. However, the sensitivity of existing methods is limited in their ability to detect rare CSF biomarkers at the treatable, early-stage of diseases. Here, we introduce novel CSF tests based on in vivo multicolor photoacoustic flow cytometry (PAFC) and ex vivo photothermal scanning cytometry. In the CSF of tumor-bearing mice, we molecularly detected in vivo circulating tumor cells (CTCs) before the development of breast cancer brain metastasis with 20-times higher sensitivity than with current assays. For the first time, we demonstrated assessing three pathways (i.e., blood, lymphatic, and CSF) of CTC dissemination, tracking nanoparticles in CSF in vivo and their imaging ex vivo. In label-free CSF samples, we counted leukocytes, erythrocytes, melanoma cells, and bacteria and imaged intracellular cytochromes, hemoglobin, melanin, and carotenoids, respectively. Taking into account the safety of PAFC, its translation for use in humans is expected to improve disease diagnosis beyond conventional detection limits. PMID:23681943

Nedosekin, Dmitry A.; Juratli, Mazen A.; Sarimollaoglu, Mustafa; Moore, Christopher L.; Rusch, Nancy J.; Smeltzer, Mark S.; Zharov, Vladimir P.; Galanzha, Ekaterina I.

2014-01-01

251

Enumeration of CD4+ T-Cells Using a Portable Microchip Count Platform in Tanzanian HIV-Infected Patients  

Microsoft Academic Search

BackgroundCD4+ T-lymphocyte count (CD4 count) is a standard method used to monitor HIV-infected patients during anti-retroviral therapy (ART). The World Health Organization (WHO) has pointed out or recommended that a handheld, point-of-care, reliable, and affordable CD4 count platform is urgently needed in resource-scarce settings.MethodsHIV-infected patient blood samples were tested at the point-of-care using a portable and label-free microchip CD4 count

SangJun Moon; Umut Atakan Gurkan; Jeffrey Blander; Wafaie W. Fawzi; Said Aboud; Ferdinand Mugusi; Daniel R. Kuritzkes; Utkan Demirci

2011-01-01

252

Amniotic fluid derived stem cells give rise to neuron-like cells without a further differentiation potential into retina-like cells  

PubMed Central

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. PMID:23862099

Hartmann, K; Raabe, O; Wenisch, S; Arnhold, S

2013-01-01

253

Automated Static Culture System Cell Module Mixing Protocol and Computational Fluid Dynamics Analysis  

NASA Technical Reports Server (NTRS)

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.

Kleis, Stanley J.; Truong, Tuan; Goodwin, Thomas J,

2004-01-01

254

Somatic Cells Count and Its Genetic Association with Milk Yield in Dairy Cattle Raised under Thai Tropical Environmental Conditions  

PubMed Central

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. PMID:25049683

Jattawa, D.; Koonawootrittriron, S.; Elzo, M. A.; Suwanasopee, T.

2012-01-01

255

Gallium-67 activity in bronchoalveolar lavage fluid in sarcoidosis  

SciTech Connect

Roentgenograms and gallium-67 scans and gallium-67 counts of BAL fluid samples, together with differential cell counts, have proved to be useful in assessing activity and lung involvement in sarcoidosis. In active pulmonary sarcoidosis gallium-67 scans are usually positive. Quantitation of gallium-67 uptake in lung scans, however, may be difficult. Because gallium-67 uptake and cell counts in BAL fluid may be correlated, we set out to investigate gallium-67 activity in BAL fluid recovered from patient of different groups. Sixteen patients with recently diagnosed and untreated sarcoidosis, nine patients with healthy lungs, and five patients with CFA were studied. Gallium-67 uptake of the lung, gallium-67 activity in the lavage fluid, SACE and LACE levels, and alpha 1-AT activity were measured. Significantly more gallium-67 activity was found in BAL fluid from sarcoidosis patients than in that from CFA patients (alpha = .001) or patients with healthy lungs (alpha = .001). Gallium-67 activity in BAL fluid could be well correlated with the number of lymphocytes in BAL fluid, but poorly with the number of macrophages. Subjects with increased levels of SACE or serum alpha 1-AT showed higher lavage gallium-67 activity than did normals, but no correlation could be established. High gallium-67 activity in lavage fluid may be correlated with acute sarcoidosis or physiological deterioration; low activity denotes change for the better. The results show that gallium-67 counts in BAL fluid reflects the intensity of gallium-67 uptake and thus of activity of pulmonary sarcoidosis.

Trauth, H.A.; Heimes, K.; Schubotz, R.; von Wichert, P.

1986-01-01

256

Genome-Wide Association Study of White Blood Cell Count in 16,388 African Americans: the Continental Origins and Genetic Epidemiology Network (COGENT)  

Microsoft Academic Search

Total white blood cell (WBC) and neutrophil counts are lower among individuals of African descent due to the common African-derived “null” variant of the Duffy Antigen Receptor for Chemokines (DARC) gene. Additional common genetic polymorphisms were recently associated with total WBC and WBC sub-type levels in European and Japanese populations. No additional loci that account for WBC variability have been

Alexander P. Reiner; Guillaume Lettre; Michael A. Nalls; Santhi K. Ganesh; Rasika Mathias; Melissa A. Austin; Eric Dean; Sampath Arepalli; Angela Britton; Zhao Chen; David Couper; J. David Curb; Charles B. Eaton; Myriam Fornage; Struan F. A. Grant; Tamara B. Harris; Dena Hernandez; Naoyuki Kamatini; Brendan J. Keating; Michiaki Kubo; Andrea LaCroix; Leslie A. Lange; Simin Liu; Kurt Lohman; Yan Meng; Emile R. Mohler; Solomon Musani; Yusuke Nakamura; Christopher J. ODonnell; Yukinori Okada; Cameron D. Palmer; George J. Papanicolaou; Kushang V. Patel; Andrew B. Singleton; Atsushi Takahashi; Hua Tang; Herman A. Taylor; Kent Taylor; Cynthia Thomson; Lisa R. Yanek; Lingyao Yang; Elad Ziv; Alan B. Zonderman; Aaron R. Folsom; Michele K. Evans; Yongmei Liu; Diane M. Becker; Beverly M. Snively; James G. Wilson

2011-01-01

257

Severe malnutrition with and without HIV1 infection in hospitalised children in Kampala, Uganda: differences in clinical features, haematological findings and CD4+ cell counts  

Microsoft Academic Search

BACKGROUND: The aim of this study was to describe the clinical features, haematological findings and CD4+ and CD8+ cell counts of severely malnourished children in relation to human immunodeficiency virus (HIV) infection. METHODS: The study was conducted in the paediatric wards of Mulago hospital, which is Uganda's national referral and teaching hospital. We studied 315 severely malnourished children (presence of

Hanifa Bachou; Thorkild Tylleskär; Robert Downing; James K Tumwine

2006-01-01

258

Equivalence of microbial biomass measures based on membrane lipid and cell wall components, adenosine triphosphate, and direct counts in subsurface aquifer sediments  

Microsoft Academic Search

An uncontaminated subsurface aquifer sediment contains a sparse microbial community consisting primarily of coccobacillary bacteria of relatively uniform size which can be counted directly with appropriate staining. The morphological simplicity and the relatively decreased cell numbers, when compared with surface soils and sediments, make the subsurface an ideal natural community with which to compare the utility of chemical measures of

David L. Balkwill; Franklin R. Leach; John T. Wilson; James F. McNabb; David C. White

1988-01-01

259

Symptomatic Illness and Low CD4 Cell Count at HIV Seroconversion as Markers of Severe Primary HIV Infection  

PubMed Central

Background The risk/benefit of initiating ART in primary HIV infection (PHI) is unclear. The benefits are more likely to outweigh the risks in patients with severe PHI. An accepted definition of severe PHI is, however, lacking. Methods CASCADE patients with HIV test interval <6 months were classified as severe and non-severe PHI based on whether the following traits were recorded in the first 6 months following seroconversion: severe specific pre-defined symptoms, central nervous system-implicated illness, and ?1, ?2 CD4<350 (and <500) cells/mm3. For each definition, we used Kaplan-Meier curves and Cox survival models to compare time to AIDS/death, censoring at the earlier of last clinic visit or 1/1/1997, when combination antiretroviral therapy (cART) became available. Results Among 1108 included patients mostly males (85%) infected through sex between men (71%), 366 were diagnosed with AIDS/died. The risk of AIDS/death was significantly higher for individuals with severe symptoms, those with ?1 CD4<350 cells/mm3 or ?2 CD4 <500 cells/mm3 in the first 6 months [aHR (95% confidence interval) 2.1 (1.4,3.2), 2.0 (1.5,2.7), and 2.3, (1.5–3.5) respectively]. Median [interquantile range] survival for patients with ?2, ?1 and no CD4<350 cells/mm3 within 6 months of seroconversion was 3.9 [2.7,6.5], 5.4 [4.5,8.4] and 8.1 [4.3,10.3] years, respectively. The diagnosis of CNS-implicated symptoms was rare and did not appear to be prognostic. Conclusion One CD4 count <350 or two <500 cells/mm3 within 6 months of seroconversion and/or severe illness in PHI may be useful early indicators of individuals at high risk of disease progression. PMID:24244330

Lodi, Sara; Fisher, Martin; Phillips, Andrew; De Luca, Andrea; Ghosn, Jade; Malyuta, Ruslan; Zangerle, Robert; Moreno, Santiago; Vanhems, Philippe; Boufassa, Faroudy; Guiguet, Marguerite; Porter, Kholoud

2013-01-01

260

Effect on quarter milk somatic cell count and antimicrobial susceptibility of Staphylococcus rostri causing intramammary infection in dairy water buffaloes.  

PubMed

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 Staph. rostri, as well as other CNS in water buffaloes. PMID:23548306

Locatelli, C; Piepers, S; De Vliegher, S; Barberio, A; Supré, K; Scaccabarozzi, L; Pisoni, G; Bronzo, V; Haesebrouck, F; Moroni, P

2013-06-01

261

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  

PubMed Central

Circulating tumor cell (CTC) counts are an established prognostic marker in metastatic prostate, breast, and colorectal cancer, and recent data suggests 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 is little 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 (range 0–515.6) and a mean of 44.7 (±95.2) HD-CTCs/mL. 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. Larger studies are warranted to investigate the prognostic value of CTC profiling in early stage lung cancer. This finding has implications for the design of larger studies examining screening, therapy, and surveillance in lung cancer patients. PMID:22307026

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

2012-01-01

262

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)

Circulating tumor cell (CTC) counts are an established prognostic marker in metastatic prostate, breast and colorectal cancer, and recent data suggest a similar role in late stage non-small cell lung cancer (NSCLC). However, due to sensitivity constraints in current enrichment-based CTC detection technologies, there are few published data about CTC prevalence rates and morphologic heterogeneity in early-stage NSCLC, or the correlation of CTCs with disease progression and their usability for clinical staging. We investigated CTC counts, morphology and aggregation in early stage, locally advanced and metastatic NSCLC patients by using a fluid-phase biopsy approach that identifies CTCs without relying on surface-receptor-based enrichment and presents them in sufficiently high definition (HD) to satisfy diagnostic pathology image quality requirements. HD-CTCs were analyzed in blood samples from 78 chemotherapy-naïve NSCLC patients. 73% of the total population had a positive HD-CTC count (>0 CTC in 1 mL of blood) with a median of 4.4 HD-CTCs mL-1 (range 0-515.6) and a mean of 44.7 (±95.2) HD-CTCs mL-1. No significant difference in the medians of HD-CTC counts was detected between stage IV (n = 31, range 0-178.2), stage III (n = 34, range 0-515.6) and stages I/II (n = 13, range 0-442.3). Furthermore, HD-CTCs exhibited a uniformity in terms of molecular and physical characteristics such as fluorescent cytokeratin intensity, nuclear size, frequency of apoptosis and aggregate formation across the spectrum of staging. Our results demonstrate that despite stringent morphologic inclusion criteria for the definition of HD-CTCs, the HD-CTC assay shows high sensitivity in the detection and characterization of both early- and late-stage lung cancer CTCs. Extensive studies are warranted to investigate the prognostic value of CTC profiling in early-stage lung cancer. This finding has implications for the design of extensive studies examining screening, therapy and surveillance in lung cancer patients.

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

263

Carryover of aflatoxin from feed to milk in dairy cows with low or high somatic cell counts.  

PubMed

Aflatoxin M1 (AFM1) residues in milk are regulated in many parts of the world and can cost dairy farmers significantly due to lost milk sales. Additionally, due to the carcinogenicity of this compound contaminated milk can be a major public health concern. Thirty-four lactating dairy cows were utilised to investigate the relationship between somatic cell counts (SCC), milk yield and conversion of dietary aflatoxin B1 (AFB1) into milk AFM1 (carryover (CO)). The AFM1 in milk increased as soon as the first milking after animal ingestion with a pattern of increment up to the observed plateau (between 7th and 12th days of AFB1 ingestion). There was a significant (P < 0.01) effect of the milk yield whereas no effect could be attributed to the SCC levels or to the milk yield × SCC interaction. Similarly, the main effect of milk yield was also observed (P < 0.01) on the total amount of AFM1 excreted during the ingestion period. Although the plasma concentration of gamma-glutamyl transferase was significantly affected by aflatoxin administration, levels of this liver enzyme were within the normal range for lactating dairy cows. The current data suggest that milk yield is the major factor affecting the total excretion of AFM1 and that SCC as an indicator of mammary gland permeability was not related to an increase in AFM1 CO. PMID:22444890

Masoero, F; Gallo, A; Moschini, M; Piva, G; Diaz, D

2007-10-01

264

Estimation of Genetic and Phenotypic Parameters for Production Traits and Somatic Cell Count for Jersey Dairy Cattle in Zimbabwe  

PubMed Central

Genetic and phenotypic parameters for production traits and somatic cell count (SCC) for Jersey dairy cattle in Zimbabwe were estimated. A total of 10986 lactation records were obtained from Zimbabwe Livestock Identification Trust, with cows calving in the period from 1996 to 2008. An ASReml program fitting an animal model was used for the analyses. Heritability estimates for milk yield, fat yield, protein yield, fat percentage, protein percentage, and Log10SCC were 0.30, 0.32, 0.33, 0.42, 0.44, and 0.08, respectively. The corresponding repeatability estimates were 0.39, 0.38, 0.39, 0.49, 0.51, and 0.16, respectively. The genetic and phenotypic correlations between different production traits ranged from ?0.86 to 0.95 and from ?0.88 to 0.98, respectively. The genetic and phenotypic correlations between production traits and Log10SCC were weak almost nonsignificantly differentl from zero. The results imply that milk traits for Jersey dairy cattle in Zimbabwe are more heritable. Therefore, these traits may be preferred by breeders as selection criteria for development of effective genetic improvement programme. PMID:23956868

Missanjo, Edward; Imbayarwo-Chikosi, Venancio; Halimani, Tinyiko

2013-01-01

265

A comparison of the evolution of the density field in perturbation theory and numerical simulations - II Counts in cells analysis  

E-print Network

We present a detailed comparison of the predictions of perturbation theory for the averaged J-point correlation functions, $\\xibar_J$, with the results of numerical simulations of gravitational clustering. We have carried out a systematic analysis of this method using ensembles of simulations with different numbers of particles, different box sizes and using different particle arrangements and clustering amplitudes in the initial conditions. We estimate $\\xibar_J$, for $J=2-10$, from moments of counts-in-cells. We find significant non-linear effects in the variance, $J=2$, even at scales as large as $R \\sim 30 \\Mpc$. Perturbation theory gives remarkable agreement at large scales, where $\\xibar_2 \\simlt 1$, with the measured hierarchical amplitudes $S_J=\\xibar_J/\\xibar_2^{J-1}$. We have followed the evolution of $\\xibar_J$ in time and find that for a change in the effective redshift of at least $\\Delta z \\simeq 2$ the amplitudes $S_J$ remain unchanged, despite the fact that the $\\xibar_J$ have evolved by large factors, $\\simeq 10^{J-1}$. We illustrate how these results can be applied to interpret the clustering in galaxy surveys and conclude that the observed hierarchical pattern in the APM is compatible with gravitational evolution in unbiased, initially Gaussian, models.

C. M. Baugh; E. Gaztanaga; G. Efstathiou

1994-08-17

266

Lingual antimicrobial peptide and lactoferrin concentrations and lactoperoxidase activity in bovine colostrum are associated with subsequent somatic cell count.  

PubMed

The present study was undertaken to examine whether potential levels of innate immune factors (lingual antimicrobial peptide (LAP), lactoferrin (LF) and lactoperoxidase (LPO)) in colostrum are associated with subsequent milk somatic cell count (SCC) in dairy cows. Quarter milk samples were collected daily for 1 week postpartum to measure LAP and LF concentrations and LPO activity. SCC in milk was determined weekly for 2 months postpartum and its correlations to concentrations of LAP and LF and LPO activity were examined. Only small variations of all immune factors were found among four udders in each individual cow, whereas there were great differences in these factors among cows. Negative correlation was detected only between LPO activity and mean and maximum SCC, whereas its relationship was not significant. LAP and LF concentrations were significantly correlated positively to mean, maximum and minimum SCC. These results suggest that the great difference in innate immune factors among animals and high LAP and LF concentrations in colostrum may be associated with subsequent high incidence of SCC increase. PMID:24001397

Isobe, Naoki; Shibata, Ayumi; Kubota, Hirokazu; Yoshimura, Yukinori

2013-11-01

267

Prognostic impact of blast cell counts in dysplastic bone marrow disorders (MDS and CMML I) with concomitant fibrosis.  

PubMed

In a retrospective study, 43 patients with dysplastic neoplasms of the bone marrow (myelodysplastic syndromes and myelodysplastic/myeloproliferative-overlap neoplasms) associated with marked (grades 2-3) fibrosis were examined. Histopathologic and morphologic findings as well as cytogenetic and molecular results were correlated with clinical endpoints. Multilineage dysplasia (34 of 43 patients, 79 %) and hypercellular bone marrow (64 %) were found in most patients. In ten of 35 patients, poor risk karyotypes according to the International Prognostic Scoring System (IPSS) were recorded. The JAK2 V617F mutation was detected in four of 30 patients (13 %), and the KIT D816V mutation was found in two of 30 patients (6 %). Patients were mainly treated with palliative drugs and best supportive care. After an observation time of 1-41 (median 21) months, ten of 43 patients (23 %) had developed a secondary acute leukemia. The median survival of all 43 patients was 21.4 months (range 1.8-88.2 months). Of all prognostic parameters examined, the blast cell count at diagnosis was found to be a most reliable and most predictive marker concerning survival and leukemia progression. This confirms previous studies in dysplastic bone marrow neoplasms without fibrosis. PMID:24241126

Machherndl-Spandl, Sigrid; Sega, W; Bösmüller, H; Germing, U; Gruber, Ch; Nachtkamp, K; Reinecke, P; Sperr, W R; Wimazal, F; Müllauer, L; Sotlar, K; Horny, H P; Tüchler, H; Valent, P; Krieger, O

2014-01-01

268

Computational fluid dynamic simulation of aggregation of deformable cells in a shear flow.  

PubMed

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. PMID:16502649

Bagchi, Prosenjit; Johnson, Paul C; Popel, Aleksander S

2005-12-01

269

Diagnosis of pulmonary histiocytosis X by immunodetection of Langerhans cells in bronchoalveolar lavage fluid.  

PubMed Central

Based on the finding that Langerhans cells and histiocytosis X cells react with the monoclonal antibody OKT6, raised against a subset of thymocytes, we used this antibody to study the cells collected by bronchoalveolar lavage (BAL) from 131 patients, including 18 with pulmonary histiocytosis X, 43 with pulmonary sarcoidosis, 67 with miscellaneous pulmonary disorders, and 3 controls. Immunofluorescence studies demonstrated the presence of OKT6-reactive cells in all patients with pulmonary histiocytosis X (mean +/- SEM, 5.29% +/- 1.14% of all cells in BAL fluid). Immunoelectron microscopic studies revealed that the cells labeled in these patients (n = 13) contained Langerhans granules. The number of fluorescent cells in the other 113 patients was significantly smaller (mean +/- SEM, 0.20% +/- 0.04% of all cells; P less than 0.001). In the 3 control patients, in the 43 patients with sarcoidosis, and in 61 of the 67 patients with miscellaneous disorders unrelated to histiocytosis X, no cells or less than 1% of the total were labeled; however, in the 6 remaining patients in this miscellaneous group, 1.3 to 2.8% of all cells in BAL were labeled. In 3 of these 6 patients, immunoelectronmicroscopic examination showed that the cells labeled by OKT6 had the general characteristics of Langerhans cells but lacked Langerhans granules. OKT3, OKT4, and OKT8 monoclonal antibodies did not stain histiocytosis X cells in BAL fluid. Images Figure 2 Figure 3 Figure 4 PMID:6372496

Chollet, S.; Soler, P.; Dournovo, P.; Richard, M. S.; Ferrans, V. J.; Basset, F.

1984-01-01

270

Microbiological screening test validation for detection of tylosin excretion in milk of cows with low and high somatic cell counts.  

PubMed

Antibiotic residues in milk above tolerance levels interfere with dairy product processing and pose potential health risks to consumers. Residue avoidance programmes include, among other components, the observance of withdrawal times indicated in label instructions. Persistence of antibiotics in milk following treatment is influenced by drug, dosage, route of administration, body weight and mammary gland health status. Compositional changes that take place during intramammary infection (IMI) can affect antibiotic excretion in milk, thus modifying milk withdrawal time. The objectives of this study were to validate sensitivity and specificity of a qualitative microbiological method (Charm AIM-96) to detect tylosin in bovine composite milk and to determine the influence of subclinical IMI in tylosin excretion following intramuscular administration. For test validation, two groups of approximately 120 cows were used; one received a single intramuscular injection of tylosin tartrate at a dose of 20 mg/kg, while the other group remained as untreated control. Test sensitivity and specificity were 100% and 94.1% respectively. To determine the influence of subclinical IMI in tylosin excretion, two groups of seven cows, one with somatic cell counts (SCC) < or =250 000 cells/ml and the other with SCC > or =900 000, were administered a single intramuscular injection of tylosin tartrate at a dose of 20 mg/kg. Milk samples were obtained every 12 h for 10 days following treatment. Milk tylosin excretion averaged between 5 and 9 days for cows with low and high SCC respectively (P < 0.0001). Compositional changes in cows with high SCC most likely affect the pharmacokinetic characteristics of tylosin, extending the presence of the antibiotic in milk, thus influencing milk withdrawal times. PMID:17359452

Litterio, N J; Calvinho, L F; Flores, M M; Tarabla, H D; Boggio, J C

2007-02-01

271

Efficacy of standard vs. extended intramammary cefquinome treatment of clinical mastitis in cows with persistent high somatic cell counts.  

PubMed

Extended duration of clinical mastitis (CM) treatment has been advocated, although results showing its higher efficacy compared with standard treatment are difficult to compare and seem conflicting. In a non-blinded, positively controlled clinical trial with systematic allocation, the efficacy of a standard, 1·5-d cefquinome treatment (ST), and an extended, 5-d intramammary cefquinome treatment (ET) were evaluated. The latter is frequently performed in cows with persistent high somatic cell count (SCC), expecting a better cure. Therefore, cows with CM immediately preceded by at least two consecutive monthly elevated SCC >200 000 cells/ml, were studied. The primary efficacy criteria were bacteriological cure (BC) and clinical cure (CC), while SCC cure was considered a secondary criterion of cure. Least square means of overall BC were not different after ET (79%, n=206) compared with ST (72%, n=203). ET, as compared with ST, improved BC of CM when caused by streptococci, specifically Streptococcus uberis. At day 1·5, only 13% of quarters showed CC, increasing significantly towards 60% at day 5, and 99% at day 14 and at day 21. No significant difference in CC was present between treatment groups. Overall SCC cure was low (22%) and not significantly different between treatment groups, but significantly higher for cases due to enterobacteriacae compared with staphylococci. In conclusion, ET with cefquinome of CM in cows with a persistent high SCC seems to be only indicated when caused by streptococci, mainly Str. uberis but shows no advantage when no information on bacteriological causes of mastitis is available. In our data, absence of CC directly after ST was not related to eventual BC. PMID:25230074

Swinkels, Jantijn M; Krömker, Volker; Lam, Theo Jgm

2014-11-01

272

Estimating test characteristics of somatic cell count to detect Staphylococcus aureus-infected dairy goats using latent class analysis.  

PubMed

The aim of this study was to estimate test properties of composite somatic cell count (cSCC) to detect subclinically Staphylococcus aureus-infected dairy goats. Staphylococcus aureus is the most prevalent major pathogen in goats, and responsible for the majority of clinical mastitis cases. Therefore, a diagnostic tool that detects subclinical Staph. aureus infections may be useful in decreasing the number of clinical cases. We collected samples from 384 animals in 4 herds for bacteriological culture and cSCC on 3 occasions in lactation: once in early lactation, once around peak lactation, and once in late lactation. Latent class models were used to estimate test properties of cSCC and bacteriological culture in the absence of a gold standard reference test under the assumption that both tests detect Staph. aureus intramammary infection. Estimates for test properties of cSCC in early lactation at a cut-off value of 1,500 × 10(3) cells/mL were 0.90 for sensitivity and 0.95 for specificity, making cSCC a useful screening tool for detection of Staph. aureus. An effect of lactation stage was observed, causing an increased sensitivity and decreased specificity in late lactation. The sensitivity of bacteriological culture was estimated to be very low in the latent class models and the models suggested that the true prevalence of Staph. aureus in dairy goat herds is much higher than what is commonly reported based on bacteriological culture. This implies that intramammary infection by Staph. aureus may be an underestimated problem in dairy goat herds, and that cSCC can be used to diagnose infected animals. PMID:21605760

Koop, G; van Werven, T; Toft, N; Nielen, M

2011-06-01

273

Quantitative Analysis of Pleural Fluid Cell-free DNA as a Tool for the Classification of Pleural Effusions  

Microsoft Academic Search

Background: Recently, much interest has been focused on the quantification of DNA in miscellaneous body fluids. In this study, the application is extended to classifying pleural effusions by measuring cell-free DNA in pleural fluid. Methods: We recruited 50 consecutive patients with pleural effusions with informed consent. Pleural fluids were centrifuged at 13 000g, with supernatants aliquoted for extraction and analysis

Michael H. M. Chan; Kai Ming Chow; Anthony T. C. Chan; Chi Bon Leung; Lisa Y. S. Chan; Katherine C. K. Chow; Ching Wan Lam; Y. M. Dennis Lo

274

Biodiversity Counts  

NSDL National Science Digital Library

This extensive collection of activities from the American Museum of Natural History offers middle school students "an exciting and creative context for involving students in the scientific process while introducing them to the rich diversity and beauty of their local ecosystem." Lesson plans, Web-based interactive activities, useful Web links, profiles of AMNH scientists and staff, and other features help students inventory and analyze the plants and arthropods found in their own neighborhoods. All activities address national science standards, and have been "field tested" in schools around the nation. Biodiversity Counts even has students develop their own exhibitions for their findings -- a great way to build science communication skills.

1998-01-01

275

In-vitro culture system for mesenchymal progenitor cells derived from waste human ovarian follicular fluid.  

PubMed

To characterize different cell populations in the human ovary, morphological and functional characteristics of cell populations collected during routine IVF procedures were studied. Cells obtained from follicular fluid grew in vitro under minimal medium conditions, without growth factor, including leukaemia-inhibiting factor. Morphological analysis revealed a heterogeneous cell population, with cells displaying a fibroblast-like, epithelial-like and also neuron-like features. Morpho-functional characteristics of fibroblast-like cells were similar to mesenchymal stem cells, and, in particular, were positive for mesenchymal stemness markers, including CD90, CD44, CD105, CD73, but negative for epithelial proteins, such as cytokeratins, CD34 and CD45 antigens. Cell proliferation activity at different times and colony-forming unit capability were evaluated, and multipotency of a subset of granulosa cells was established by in-vitro differentiation studies (e.g. osteogenic, chondrogenic and adipogenic differentiation). This study suggests that cells provided by mesenchymal plasticity can be easily isolated by waste follicular fluid, avoiding scraping of human ovaries, and cultivated in minimal conditions. Successful growth of such progenitor cells on three-dimensional cryogel scaffold provides the basis for future developments in tissue engineering. This culture system may be regarded as an experimental model in which biological behaviour is not influenced by specific growth factors. PMID:25131558

Riva, Federica; Omes, Claudia; Bassani, Roberto; Nappi, Rossella E; Mazzini, Giuliano; Icaro Cornaglia, Antonia; Casasco, Andrea

2014-10-01

276

Mastitis Is Associated with Increased Free Fatty Acids, Somatic Cell Count, and Interleukin-8 Concentrations in Human Milk  

PubMed Central

Abstract Background Research in bovine lactation has demonstrated that milk produced by a mammary gland displaying inflammation-based symptoms of mastitis has increased levels of free fatty acids (FFAs) compared with milk produced by a contralateral asymptomatic gland. However, the effects of mastitis on lipid classes in milk have not been investigated in humans. Methods The study described here compared milk collected from the symptomatic breast of women with mastitis (n=14) with that collected from the contralateral asymptomatic breast to determine if mastitis caused alterations in the quantity of total lipids, FFAs, and phospholipids (PLs), as well as the fatty acid profiles of these lipid classes. To assess their efficacy as biomarkers of mastitis, samples were also analyzed for selected markers of local inflammation: sodium, somatic cell count (SCC), and interleukin-8 (IL-8). Results FFAs were higher in milk from the mastitic breast compared with that from the healthy breast (1.31 vs. 1.07±0.10?g/100?g of lipid, p<0.05). Similarly, SCC and IL-8 were elevated roughly 10-fold in milk from mastitic breasts, compared with milk from healthy breasts, and sodium tended to be higher in milk from mastitic breasts (p<0.10). However, there were no differences in total lipid, PLs, or fatty acid profiles within each lipid class. Conclusions In summary, mastitis is associated with increased lipolysis in the human breast but not alterations in milk fat synthesis, as evidenced by a lack of alteration in total milk lipids. Additionally, these results indicate that SCC and IL-8 may be better indicators of mammary inflammation than sodium content. PMID:22283504

Hunt, Katherine M.; Williams, Janet E.; Shafii, Bahman; Hunt, Martha K.; Behre, Rebecca; Ting, Robert; McGuire, Michelle K.

2013-01-01

277

CD8+ T-cell counts: an early predictor of risk and mortality in critically ill immunocompromised patients with invasive pulmonary aspergillosis  

PubMed Central

Introduction Critically ill immunocompromised (CIIC) patients with pulmonary infection are a population at high risk for invasive pulmonary aspergillosis (IPA). The host defenses are important factors to consider in determining the risk and outcome of infection. Quantification of changes in the status of host immunity could be valuable for clinical diagnosis and outcome prediction. Methods We evaluated the quantitative changes in key humoral and cellular parameters in CIIC patients with pulmonary infection and their potential influence on the risk and prognosis of IPA. We monitored the evolution of these parameters in 150 CIIC patients with pulmonary infection on days 1, 3 and 10 (D1, D3 and D10) following ICU admission. The primary outcome was 28-day mortality. Follow-up included 60- and 90-day mortality. Results Among the 150 CIIC patients included in this study, 62 (41.3%) had microbiological evidence of IPA. Compared with patients without IPA, CD3+, CD8+, CD28+CD4+ and CD28+CD8+ CD28+CD8+ T-cell counts (D1, D3 and D10) and B-cell counts (D1 and D3) were significantly reduced in patients with IPA (P?cell counts were independent predictors of IPA in CIIC patients. Receiver operating characteristic analysis of immune parameters predicting 28-day mortality revealed area under the curve values of 0.82 (95% CI 0.71 to 0.92), 0.94 (95% CI 0.87 to 0.99), and 0.94 (95% CI 0.85 to 0.99) for CD8+ T-cell counts (D1, D3 and D10, respectively) and 0.84 (95% CI 0.75 to 0.94), 0.92 (95% CI 0.85 to 0.99) and 0.90 (95% CI 0.79 to 0.99) for CD28+CD8+ T-cell counts (D1, D3 and D10, respectively). Kaplan-Meier survival analysis provided evidence that CD8+ and CD28+CD8+ T-cell counts (<149.5 cells/mm3 and <75 cells/mm3, respectively) were associated with early mortality in CIIC patients with IPA (logrank test; P?cell counts were significantly lower in CIIC patients with IPA than in non-IPA patients. Lower CD8+ and CD28+CD8+ T-cell counts in CIIC patients with pulmonary infection were associated with higher risk and early mortality in IPA and may be valuable for clinical diagnosis and outcome prediction. PMID:23883548

2013-01-01

278

Total and Differential White Blood Cell Counts Predict Eight-Year Incident Stoke in Elderly Japanese-American Men: The Honolulu Heart Program  

PubMed Central

Background: Previous studies have found that a higher white blood cell (WBC) count is associated with incident stroke. There have been few studies examining differential WBC counts in elderly or Asian populations. We studied the association between total and differential WBC counts and incident stroke in an older Asian population. Methods: The Honolulu Heart Program is a prospective population-based study of cardiovascular diseases in Japanese-American men that started in 1965. At exam 4 (1991–93), 3,741 men ages 71–93 years participated, and total and differential WBC counts were measured in 3,569 men using a Coulter counter machine. Data on incident stroke (all strokes [ALL-CVA], thromboembolic [TE-CVA] and hemorrhagic [HEM-CVA]) were available through December 1999 (8 years follow-up) from a comprehensive hospital surveillance system. After excluding 227 subjects with prevalent stroke, 3,342 subjects were divided into quartiles of total WBC, neutrophil (segmented and band), granulocyte (neutrophil, eosinophil and basophil), lymphocyte, and monocyte counts for separate analyses. Results: Age-adjusted incident ALL-CVA rates increased significantly with total WBC quartiles (7.68, 9.04, 9.26, 14.1, per 1,000 person years follow-up, respectively, P = .0014). Relative risks for ALL-CVA for each quartile of total and differential WBC counts were obtained using Cox proportional hazards, using the lowest quartile as the reference group. After full adjustment including age, cardiovascular risk factors, fibrinogen, prevalent CHD, cancer or COPD, and aspirin/NSAID use, the relative risks in the highest quartiles of total WBC, neutrophil, and granulocyte counts were 1.63 (95%CI = 1.05–2.54, P = .03), 2.19 (95%CI = 1.41–3.39, P < .001) and 1.91 (95%CI = 1.25–2.92, P = .003), respectively. These significant associations were also seen for TE-CVA, but not for HEM-CVA. No significant associations were found between lymphocyte or monocyte counts and incident stroke or subtypes. Conclusions: In elderly Japanese-American men, higher total WBC, neutrophil, and granulocyte counts were independent predictors of overall stroke, as well as thromboembolic stroke. Further studies are needed to establish cut-points and treatment options.

Ross, G Webster; Chen, Randi; Bell, Christina; Willcox, Bradley; Abbott, Robert; Launer, Lenore; Kaya, Brock; Masaki, Kamal

2014-01-01

279

Computational and Experimental Models of Cancer Cell Response to Fluid Shear Stress  

PubMed Central

It has become evident that mechanical forces play a key role in cancer metastasis, a complex series of steps that is responsible for the majority of cancer-related deaths. One such force is fluid shear stress, exerted on circulating tumor cells by blood flow in the vascular microenvironment, and also on tumor cells exposed to slow interstitial flows in the tumor microenvironment. Computational and experimental models have the potential to elucidate metastatic behavior of cells exposed to such forces. Here, we review the fluid-generated forces that tumor cells are exposed to in the vascular and tumor microenvironments, and discuss recent computational and experimental models that have revealed mechanotransduction phenomena that may play a role in the metastatic process. PMID:23467856

Mitchell, Michael J.; King, Michael R.

2013-01-01

280

Therapeutic outcomes of transplantation of amniotic fluid-derived stem cells in experimental ischemic stroke.  

PubMed

Accumulating preclinical evidence suggests the use of amnion as a source of stem cells for investigations of basic science concepts related to developmental cell biology, but also for stem cells' therapeutic applications in treating human disorders. We previously reported isolation of viable rat amniotic fluid-derived stem (AFS) cells. Subsequently, we recently reported the therapeutic benefits of intravenous transplantation of AFS cells in a rodent model of ischemic stroke. Parallel lines of investigations have provided safety and efficacy of stem cell therapy for treating stroke and other neurological disorders. This review article highlights the need for investigations of mechanisms underlying AFS cells' therapeutic benefits and discusses lab-to-clinic translational gating items in an effort to optimize the clinical application of the cell transplantation for stroke. PMID:25165432

Tajiri, Naoki; Acosta, Sandra; Portillo-Gonzales, Gabriel S; Aguirre, Daniela; Reyes, Stephanny; Lozano, Diego; Pabon, Mibel; Dela Peña, Ike; Ji, Xunming; Yasuhara, Takao; Date, Isao; Solomita, Marianna A; Antonucci, Ivana; Stuppia, Liborio; Kaneko, Yuji; Borlongan, Cesar V

2014-01-01

281

Therapeutic outcomes of transplantation of amniotic fluid-derived stem cells in experimental ischemic stroke  

PubMed Central

Accumulating preclinical evidence suggests the use of amnion as a source of stem cells for investigations of basic science concepts related to developmental cell biology, but also for stem cells’ therapeutic applications in treating human disorders. We previously reported isolation of viable rat amniotic fluid-derived stem (AFS) cells. Subsequently, we recently reported the therapeutic benefits of intravenous transplantation of AFS cells in a rodent model of ischemic stroke. Parallel lines of investigations have provided safety and efficacy of stem cell therapy for treating stroke and other neurological disorders. This review article highlights the need for investigations of mechanisms underlying AFS cells’ therapeutic benefits and discusses lab-to-clinic translational gating items in an effort to optimize the clinical application of the cell transplantation for stroke. PMID:25165432

Tajiri, Naoki; Acosta, Sandra; Portillo-Gonzales, Gabriel S.; Aguirre, Daniela; Reyes, Stephanny; Lozano, Diego; Pabon, Mibel; Dela Pena, Ike; Ji, Xunming; Yasuhara, Takao; Date, Isao; Solomita, Marianna A.; Antonucci, Ivana; Stuppia, Liborio; Kaneko, Yuji; Borlongan, Cesar V.

2014-01-01

282

Influence of Fluid Velocity and Cell Concentration on the Transport of  

E-print Network

Influence of Fluid Velocity and Cell Concentration on the Transport of Motile and Nonmotile Bacteria in Porous Media T E R R I A . C A M E S A N O A N D B R U C E E . L O G A N * Department of Civil and, therefore, should result in increased colloid retention in porous media. However, for motile

283

Automatic counting and positioning of 5-bromo-2-deoxyuridine (BrdU) positive cells in cortical layers of rat brain slices.  

PubMed

5-Bromo-2-deoxyuridine (BrdU) staining is often used to evaluate cortical layer formation during mammalian brain development. This method allows the quantification of newly generated cells and therefore the study of the effects of xenobiotics or genetic factors on proliferation, cell death and migration behavior in a quantitative manner. However, these endpoints are generally assessed by time-consuming manual evaluation. In the present work, we introduce a novel procedure to identify and quantify BrdU(+) cells within cortical layers, using the commercially available vHCS-Scan V.6.3.1 software to identify BrdU(+) cell coordinates and the novel program 'BrdeLuxe' to define cortical layers and quantitatively assign BrdU(+) cells to them. This procedure is compared to BrdU(+) cell counting with the freeware 'ImageJ' in respect to the manual evaluation, all by two different researchers. BrdeLuxe shows high accuracy and precision for the determination of total number of BrdU(+) cells compared to the manual counting, while ImageJ does not reach such results. Accuracy and precision are also higher for employing the BrdeLuxe program to evaluate the percentage of BrdU(+) cells per brain layer compared to ImageJ. In terms of running time, BrdeLuxe is the fastest method of the three making it more suitable for multiple brain slices analyses. PMID:24572144

Schmuck, Martin; Temme, Thomas; Heinz, Sabrina; Baksmeier, Christine; Mosig, Axel; Colomina, M Teresa; Barenys, Marta; Fritsche, Ellen

2014-07-01

284

Applications of Amniotic Membrane and Fluid in Stem Cell Biology and Regenerative Medicine  

PubMed Central

The amniotic membrane (AM) and amniotic fluid (AF) have a long history of use in surgical and prenatal diagnostic applications, respectively. In addition, the discovery of cell populations in AM and AF which are widely accessible, nontumorigenic and capable of differentiating into a variety of cell types has stimulated a flurry of research aimed at characterizing the cells and evaluating their potential utility in regenerative medicine. While a major focus of research has been the use of amniotic membrane and fluid in tissue engineering and cell replacement, AM- and AF-derived cells may also have capabilities in protecting and stimulating the repair of injured tissues via paracrine actions, and acting as vectors for biodelivery of exogenous factors to treat injury and diseases. Much progress has been made since the discovery of AM and AF cells with stem cell characteristics nearly a decade ago, but there remain a number of problematic issues stemming from the inherent heterogeneity of these cells as well as inconsistencies in isolation and culturing methods which must be addressed to advance the field towards the development of cell-based therapies. Here, we provide an overview of the recent progress and future perspectives in the use of AM- and AF-derived cells for therapeutic applications. PMID:23093978

Rennie, Kerry; Gruslin, Andree; Hengstschlager, Markus; Pei, Duanqing; Cai, Jinglei; Nikaido, Toshio; Bani-Yaghoub, Mahmud

2012-01-01

285

Quantification and significance of fluid shear stress field in biaxial cell stretching device  

Microsoft Academic Search

A widely used commercially available system for the investigation of mechanosensitivity applies a biaxial strain field to\\u000a cells cultured on a compliant silicone substrate membrane stretched over a central post. As well as intended substrate strain,\\u000a this device also provides a fluid flow environment for the cultured cells. In order to interpret the relevance of experiments\\u000a using this device to

Mark S. Thompson; Stuart R. Abercrombie; Claus-Eric Ott; Friederike H. Bieler; Georg N. Duda; Yiannis Ventikos

2011-01-01

286

Pore Network Modeling of Capillary Pressure -- Saturation Relations Through Retention Cells and Equilibrium Fluid Distributions in Long Columns  

Microsoft Academic Search

Standard methods for experimentally determining capillary pressure -- saturation relations of a porous medium use retention cells: a small cell filled with the porous medium is connected to two pressure reservoirs for the two given fluids. Then, the saturation is recorded as the pressure difference is changed. Conversely, one can infer capillary pressure -- saturation relations by analyzing equilibrium fluid

R. Glantz; M. Hilpert

2009-01-01

287

Demonstration of mast cell chemotactic activity in bronchoalveolar lavage fluid collected from asthmatic patients before and during pollen season  

Microsoft Academic Search

Background: Mast cells are versatile effector cells of primary importance in asthma and airway inflammation. During inflammation mast cells accumulate in the bronchial epithelium. The mechanism for this increase in mast cell number has not been defined. Objectives: The aim of this study was to examine the presence of mast cell chemotactic activity in bronchoalveolar lavage (BAL) fluid taken before

Niclas Olsson; Sabina Rak; Gunnar Nilsson

2000-01-01

288

Fluid flow through a high cell density fluidized-bed during centrifugal bioreactor culture.  

PubMed

An increasing demand for products such as tissues, proteins, and antibodies from mammalian cell suspension cultures is driving interest in increasing production through high-cell density bioreactors. The centrifugal bioreactor (CCBR) retains cells by balancing settling forces with surface drag forces due to medium throughput and is capable of maintaining cell densities above 10(8) cells/mL. This article builds on a previous study where the fluid mechanics of an empty CCBR were investigated showing fluid flow is nonuniform and dominated by Coriolis forces, raising concerns about nutrient and cell distribution. In this article, we demonstrate that the previously reported Coriolis forces are still present in the CCBR, but masked by the presence of cells. Experimental dye injection observations during culture of 15 microm hybridoma cells show a continual uniform darkening of the cell bed, indicating the region of the reactor containing cells is well mixed. Simulation results also indicate the cell bed is well mixed during culture of mammalian cells ranging in size from 10 to 20 microm. However, simulations also allow for a slight concentration gradient to be identified and attributed to Coriolis forces. Experimental results show cell density increases from 0.16 to 0.26 when centrifugal force is doubled by increasing RPM from 650 to 920 at a constant inlet velocity of 6.5 cm/s; an effect also observed in the simulation. Results presented in this article indicate cells maintained in the CCBR behave as a high-density fluidized bed of cells providing a homogeneous environment to ensure optimal growth conditions. PMID:20205172

Detzel, Christopher J; Van Wie, Bernard J; Ivory, Cornelius F

2010-01-01

289

Very late activation antigens on rheumatoid synovial fluid T lymphocytes. Association with stages of T cell activation.  

PubMed Central

Lymphocytes from the synovial fluid of eight out of eight rheumatoid arthritis (RA) patients had elevated very late activation antigen-1 (VLA-1) expression (10-36% positive cells), whereas peripheral blood lymphocytes (PBL) from RA patients and healthy controls had low VLA-1 expression (0-6% positive cells). During 1-2 wk of in vitro culture, VLA-1 increased on synovial fluid cells but remained low on PBL. In comparison, the interleukin 2 receptor (IL-2 R) was less prominent than VLA-1 on fresh synovial fluid cells, did not increase on cultured synovial fluid T cells, but did increase greatly on cultured PBL. The mitogen PHA reversed or prevented the appearance of VLA-1+, IL-2 R- synovial fluid cells during in vitro culture, thus giving IL-2 R+, VLA-1- cells. These results emphasize that VLA-1+ SF cells are different from resting cells or IL-2 R+ activated PBL T cells, and VLA-1 on synovial fluid T cells may be incompatible with mitogen stimulation. In addition, the VLA-2 heterodimer (165,000/130,000 relative molecular mass [Mr]) was regulated opposite to the VLA-1 heterodimer (130,000/210,000 Mr) on synovial lymphocytes, and thus the VLA-1/VLA-2 ratio is another indicator of the stage of T cell activation. Images PMID:3018043

Hemler, M E; Glass, D; Coblyn, J S; Jacobson, J G

1986-01-01

290

Renal ?-intercalated cells maintain body fluid and electrolyte balance.  

PubMed

Inactivation of the B1 proton pump subunit (ATP6V1B1) in intercalated cells (ICs) leads to type I distal renal tubular acidosis (dRTA), a disease associated with salt- and potassium-losing nephropathy. Here we show that mice deficient in ATP6V1B1 (Atp6v1b1-/- mice) displayed renal loss of NaCl, K+, and water, causing hypovolemia, hypokalemia, and polyuria. We demonstrated that NaCl loss originated from the cortical collecting duct, where activity of both the epithelial sodium channel (ENaC) and the pendrin/Na(+)-driven chloride/bicarbonate exchanger (pendrin/NDCBE) transport system was impaired. ENaC was appropriately increased in the medullary collecting duct, suggesting a localized inhibition in the cortex. We detected high urinary prostaglandin E2 (PGE2) and ATP levels in Atp6v1b1-/- mice. Inhibition of PGE2 synthesis in vivo restored ENaC protein levels specifically in the cortex. It also normalized protein levels of the large conductance calcium-activated potassium channel and the water channel aquaporin 2, and improved polyuria and hypokalemia in mutant mice. Furthermore, pharmacological inactivation of the proton pump in ?-ICs induced release of PGE2 through activation of calcium-coupled purinergic receptors. In the present study, we identified ATP-triggered PGE2 paracrine signaling originating from ?-ICs as a mechanism in the development of the hydroelectrolytic imbalance associated with dRTA. Our data indicate that in addition to principal cells, ICs are also critical in maintaining sodium balance and, hence, normal vascular volume and blood pressure. PMID:24051376

Gueutin, Victor; Vallet, Marion; Jayat, Maximilien; Peti-Peterdi, Janos; Cornière, Nicolas; Leviel, Françoise; Sohet, Fabien; Wagner, Carsten A; Eladari, Dominique; Chambrey, Régine

2013-10-01

291

Fluid Flow Mechanotransduction in Vascular Smooth Muscle Cells and Fibroblasts  

PubMed Central

Understanding how vascular wall endothelial cells (ECs), smooth muscle cells (SMCs), and fibroblasts (FBs) sense and transduce the stimuli of hemodynamic forces (shear stress, cyclic strain, and hydrostatic pressure) into intracellular biochemical signals is critical to prevent vascular disease development and progression. ECs lining the vessel lumen directly sense alterations in blood flow shear stress and then communicate with medial SMCs and adventitial FBs to regulate vessel function and disease. Shear stress mechanotransduction in ECs has been extensively studied and reviewed. In the case of endothelial damage, blood flow shear stress may directly act on the superficial layer of SMCs and transmural interstitial flow may be elevated on medial SMCs and adventitial FBs. Therefore, it is also important to investigate direct shear effects on vascular SMCs as well as FBs. The work published in the last two decades has shown that shear stress and interstitial flow have significant influences on vascular SMCs and FBs. This review summarizes work that considered direct shear effects on SMCs and FBs and provides the first comprehensive overview of the underlying mechanisms that modulate SMC secretion, alignment, contraction, proliferation, apoptosis, differentiation, and migration in response to 2-dimensional (2D) laminar, pulsatile, and oscillating flow shear stresses and 3D interstitial flow. A mechanistic model of flow sensing by SMCs is also provided to elucidate possible mechanotransduction pathways through surface glycocalyx, integrins, membrane receptors, ion channels, and primary cilia. Understanding flow-mediated mechanotransduction in SMCs and FBs and the interplay with ECs should be helpful in exploring strategies to prevent flow-initiated atherosclerosis and neointima formation and has implications in vascular tissue engineering. PMID:21479754

Shi, Zhong-Dong; Tarbell, John M.

2011-01-01

292

A white blood cell count in the normal concentration range is independently related to cardiorespiratory fitness in apparently healthy Korean men  

Microsoft Academic Search

Despite the documented health benefits of physical activity, the mechanism whereby physical activity prevents cardiovascular disease is incompletely understood. In the present study, we investigated the relationship between white blood cell (WBC) count and cardiorespiratory fitness (V?o2max) after adjusting for several well-known cardiovascular risk factors. Subjects who visited our health promotion center for a medical checkup and treadmill test (n

Dong-Jun Kim; Jung-Hyun Noh; Byung-Wan Lee; Yoon-Ho Choi; Jae-Hoon Jung; Yong-Ki Min; Myung-Shik Lee; Moon-Kyu Lee; Kwang-Won Kim

2005-01-01

293

White Blood Cell Count and Incidence of Coronary Heart Disease and Ischemic Stroke and Mortality from Cardiovascular Disease in African American and White Men and Women  

Microsoft Academic Search

) had 1.9 times the risk of incident coronary heart disease (95% confidence interval (CI): 1.19, 3.09), 1.9 times the risk of incident ischemic stroke (95% CI: 1.03, 3.34), and 2.3 times the risk of cardiovascular disease mortality (95% CI: 1.38, 3.72) as their counterparts in the lowest quartile of WBC count (<4,800 cells\\/mm 3 ). These associations were similar

Chong Do Lee; Aaron R. Folsom; F. Javier Nieto; Lloyd E. Chambless; Eyal Shahar; Douglas A. Wolfe

294

Reticulocyte count  

MedlinePLUS

... red blood cells being destroyed earlier than normal ( hemolytic anemia ) Bleeding Blood disorder in a fetus or newborn known as erythroblastosis fetalis Kidney disease, with increased production of a hormone called erythropoietin ...

295

Joint estimation of genetic parameters for test-day somatic cell count and mastitis in the United Kingdom.  

PubMed

Genetic parameters were estimated in a joint analysis of log(e)-transformed somatic cell count (TSCC) with either mastitis as a binary trait (MAS) or the number of mastitis cases (NMAS) in Holstein-Friesian cows for the first 3 lactations using a random regression model. In addition, a multi-trait analysis of MAS and NMAS was also implemented. There were 67,175, 30,617, and 16,366 cows with records for TSCC, MAS, and NMAS in lactations 1, 2, and 3, respectively. The frequency of MAS was 14, 20, and 25% in lactations 1, 2, and 3 respectively. The model for TSCC included herd-test-day, age at calving and month of calving, fixed lactation curves nested with calving year groups, and random regressions with Legendre polynomials of order 2 for animal and permanent environmental effects. The model for MAS and NMAS included fixed herd-year-season, age at calving and month of calving, and random animal and permanent environmental effects. All analyses were carried out using Gibbs sampling. Estimates of mean daily heritability averaged over a 305-d lactation were 0.11, 0.14, and 0.15 for TSCC for lactations 1, 2, and 3, respectively. Corresponding heritability estimates for MAS were 0.05, 0.07, and 0.09. The heritabilities for NMAS were similar at 0.06, 0.07, and 0.12, respectively, for lactations 1, 2, and 3. The genetic correlations between lactations 1 and 2, 1 and 3, and 2 and 3 were 0.75, 0.64, and 0.92 for computed 305-d lactation TSCC; 0.55, 0.48, and 0.89 for MAS; and 0.62, 0.42, and 0.85 for NMAS, respectively. The genetic correlations between MAS and TSCC were positive and generally moderate to high. The genetic correlations between computed 305-d lactation TSCC and MAS were 0.53, 0.61, and 0.68 in lactations 1, 2, and 3, respectively. Similar corresponding genetic correlations were obtained between computed 305-d lactation TSCC and NMAS in the respective parities. Mastitis as a binary trait and NMAS in the same lactation were very highly correlated and were genetically the same trait. It is intended that the new parameters will be used in setting up a national evaluation system for the joint analysis of TSCC and MAS. PMID:22818477

Mrode, R; Pritchard, T; Coffey, M; Wall, E

2012-08-01

296

Experiment 7: The Geophysical Fluid Flow Cell Experiment on USML-2  

NASA Technical Reports Server (NTRS)

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 non-axisymmetric 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.

Hart, John E.; Ohlsen, Daniel R.; Kittleman, Scott; Leslie, Fred W.; Miller, Timothy L.

1998-01-01

297

The interaction between a solid body and viscous fluid by marker-and-cell method  

NASA Technical Reports Server (NTRS)

A computational method for solving nonlinear problems relating to impact and penetration of a rigid body into a fluid type medium is presented. The numerical techniques, based on the Marker-and-Cell method, gives the pressure and velocity of the flow field. An important feature in this method is that the force and displacement of the rigid body interacting with the fluid during the impact and sinking phases are evaluated from the boundary stresses imposed by the fluid on the rigid body. A sample problem of low velocity penetration of a rigid block into still water is solved by this method. The computed time histories of the acceleration, pressure, and displacement of the block show food agreement with experimental measurements. A sample problem of high velocity impact of a rigid block into soft clay is also presented.

Cheng, R. Y. K.

1976-01-01

298

Cause-specific mortality among HIV-infected individuals, by CD4+ cell count at HAART initiation, compared with HIV-uninfected individuals  

PubMed Central

Objectives To compare the proportion, timing and hazards of non-AIDS death and AIDS death among men and women who initiated HAART at different CD4+ cell counts to mortality risks of HIV-uninfected persons with similar risk factors. Design Prospective cohort studies. Methods We used parametric mixture models to compare proportions of AIDS and non-AIDS mortality and ages at death, and multivariable Cox models to compare cause-specific hazards of mortality, across levels of CD4+ cell count at HAART initiation (?200 cells/?l: ‘late’, 201–350 cells/?l: ‘intermediate’, >350 cells/?l: ‘early’) and with HIV-uninfected individuals from the Multicenter AIDS Cohort Study and the Women’s Interagency HIV Study. We used multiple imputation methods to address lead-time bias in sensitivity analysis. Results Earlier initiators were more likely to die of non-AIDS causes (early: 78%, intermediate: 74%, late: 49%), and at older ages (median years 72, 69, 66), relative to later initiators. Estimated median ages at non-AIDS death for each CD4+ cell count category were lower than that estimated for the HIV-uninfected group (75 years). In multivariable analysis, non-AIDS death hazard ratios relative to early initiators were 2.15 for late initiators (P < 0.01) and 1.66 for intermediate initiators (P = 0.01); AIDS death hazard ratios were 3.26 for late initiators (P <0.01) and 1.20 for intermediate initiators (P = 0.28). Strikingly, the adjusted hazards for non-AIDS death among HIV-uninfected individuals and early initiators were nearly identical (hazard ratio 1.01). Inferences were unchanged after adjustment for lead-time bias. Conclusion Results suggest the possibility of reducing the risk of non-AIDS mortality among HIV-infected individuals to approximate that faced by comparable HIV-uninfected individuals. PMID:24105030

Wada, Nikolas; Jacobson, Lisa P.; Cohen, Mardge; French, Audrey; Phair, John; Munoz, Alvaro

2014-01-01

299

Human amniotic fluid promotes retinal pigmented epithelial cells' trans-differentiation into rod photoreceptors and retinal ganglion cells.  

PubMed

To evaluate the effect of human amniotic fluid (HAF) on retinal pigmented epithelial cells growth and trans-differentiation into retinal neurons, retinal pigmented epithelium (RPE) cells were isolated from neonatal human cadaver eye globes and cultured in Dulbecco's modified Eagle's medium-F12 supplemented with 10% fetal bovine serum (FBS). Confluent monolayer cultures were trypsinized and passaged using FBS-containing or HAF-containing media. Amniotic fluid samples were received from pregnant women in the first trimester of gestation. Cell proliferation and death enzyme-linked immunosorbent assays were performed to assess the effect of HAF on RPE cell growth. Trans-differentiation into rod photoreceptors and retinal ganglion cells was also studied using immunocytochemistry and real-time polymerase chain reaction techniques. Primary cultures of RPE cells were successfully established under FBS-containing or HAF-containing media leading to rapid cell growth and proliferation. When RPE cells were moved to in vitro culture system, they began to lose their differentiation markers such as pigmentation and RPE65 marker and trans-differentiated neural-like cells followed by spheroid colonies pertaining to stem/progenitor cells were morphologically detected. Immunocytochemistry (ICC) analysis of HAF-treated cultures showed a considerable expression of Rhodopsin gene (30% Rhodopsin-positive cells) indicating trans-differentiation of RPE cells to rod photoreceptors. Real-time polymerase chain reaction revealed an HAF-dose-dependant expression of Thy-1 gene (RGC marker) and significant promoting effect of HAF on RGCs generation. The data presented here suggest that HAF possesses invaluable stimulatory effect on RPE cells growth and trans-differentiation into retinal neurons. It can be regarded as a newly introduced enriched supplement in serum-free kinds of media used in neuro-retinal regeneration studies. PMID:21142973

Ghaderi, Shima; Soheili, Zahra-Soheila; Ahmadieh, Hamid; Davari, Maliheh; Jahromi, Fatemeh Sanie; Samie, Shahram; Rezaie-Kanavi, Mozhgan; Pakravesh, Jalil; Deezagi, Abdolkhalegh

2011-09-01

300

[Cell count and Schalm test results of milk from dairy sheep with healthy udders during the course of a complete lactation].  

PubMed

The somatic cell count (SCC) was determined in 28 milk sheep with normal udder health using the California Mastitis Test (CMT) and a fluoro-opto-electronic cell counter (Partec Counter). The examinations were made at two week intervals throughout the lactation period. 92% of the premilking samples had a negative CMT reaction and in 0.5% of the samples the reaction was distinctly positive. In 95% of the premilking samples the SCC showed less than 200,000/ml and in 95% of the individual total milkings it was less than 350,000/ml. Elevated SCC were common towards the end of lactation when milk yield was low. PMID:2047833

Regi, G; Honegger, R; Büchi, S; Segessemann, V; Rüsch, P

1991-01-01

301

An altered repertoire of T cell receptor V gene expression by rheumatoid synovial fluid T lymphocytes.  

PubMed Central

The pattern of T cell receptor V gene expression by lymphocytes from rheumatoid synovial fluid and paired peripheral blood samples was compared using a polymerase chain reaction (PCR)-based assay. Eight rheumatoid arthritis (RA) patients who had varying durations of disease (from 2 to 20 years) were studied. In all patients there was evidence of a different pattern of V gene expression between the two compartments. Significantly increased expression of at least one V alpha or V beta gene family by synovial fluid T cells was observed in all the patients studied. Three different V alpha (V alpha 10, 15 and 18) and three V beta (V beta 4, 5 and 13) families were commonly elevated. Sequencing of synovial V beta transcripts demonstrated that the basis of increased expression of selected V gene families in the synovial fluid was due to the presence of dominant clonotypes within those families, which constituted up to 53% of the sequences isolated from one particular synovial V gene family. There were considerable differences in the NDJ sequences found in synovial and peripheral blood T cell receptor (TCR) transcripts of the same V beta gene family. These data suggest that the TCR repertoire in the two compartments differs, and that antigen-driven expansion of particular synovial T cell populations is a component of rheumatoid synovitis, and is present in all stages of the disease. PMID:1458680

Lunardi, C; Marguerie, C; So, A K

1992-01-01

302

Cerebrospinal fluid B cells from Multiple Sclerosis patients are subject to normal germinal center selection  

PubMed Central

Previous findings from our laboratory demonstrated that some clonally expanded cerebrospinal fluid (CSF) B cells from MS patients exhibit diminished mutation targeting patterns in comparison to typical B cells selected in the context of germinal centers (GCs). In order to determine whether the overall CSF B cell repertoires adhered to mutation patterns typical of GC-selected B cells, we analyzed the immunoglobulin repertoires from CSF B cells of 8 MS patients for mutation characteristics typical of GC-derived B cells. Mutation targeting was preserved. Thus, clonal expansion of some CSF B cells may occur independently of GC, but the CSF B cell pool is governed by typical GC selection. Interestingly, the heavy chain CDR3’s of CSF B cells from MS patients had a net acidic charge, similar to GC-derived B cells, but a tendency towards longer CDR3’s, consistent with autoreactive B cells. How these findings may support current hypotheses regarding the origin of CSF B cells is discussed. PMID:17169437

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

2007-01-01

303

High plasma fibrinogen concentration and platelet count unfavorably impact survival in non-small cell lung cancer patients with brain metastases  

PubMed Central

High expression of fibrinogen and platelets are often observed in non–small cell lung cancer (NSCLC) patients with local regional or distant metastasis. However, the role of these factors remains unclear. The aims of this study were to evaluate the prognostic significance of plasma fibrinogen concentration and platelet count, as well as to determine the overall survival of NSCLC patients with brain metastases. A total of 275 NSCLC patients with brain metastasis were enrolled into this study. Univariate analysis showed that high plasma fibrinogen concentration was associated with age?65 years (P = 0.011), smoking status (P = 0.009), intracranial symptoms (P = 0.022), clinical T category (P = 0.010), clinical N category (P = 0.003), increased partial thromboplastin time (P < 0.001), and platelet count (P < 0.001). Patients with low plasma fibrinogen concentration demonstrated longer overall survival compared with those with high plasma fibrinogen concentration (median, 17.3 months versus 11.1 months; P?0.001). A similar result was observed for platelet counts (median, 16.3 months versus 11.4 months; P = 0.004). Multivariate analysis showed that both plasma fibrinogen concentration and platelet count were independent prognostic factors for NSCLC with brain metastases (R2 = 1.698, P < 0.001 and R2 = 1.699, P < 0.001, respectively). Our results suggest that high plasma fibrinogen concentration and platelet count indicate poor prognosis for NSCLC patients with brain metastases. Thus, these two biomarkers might be independent prognostic predictors for this subgroup of NSCLC patients. PMID:23958057

Zhu, Jian-Fei; Cai, Ling; Zhang, Xue-Wen; Wen, Yin-Sheng; Su, Xiao-Dong; Rong, Tie-Hua; Zhang, Lan-Jun

2014-01-01

304

High plasma fibrinogen concentration and platelet count unfavorably impact survival in non-small cell lung cancer patients with brain metastases.  

PubMed

High expression of fibrinogen and platelets are often observed in non-small cell lung cancer (NSCLC) patients with local regional or distant metastasis. However, the role of these factors remains unclear. The aims of this study were to evaluate the prognostic significance of plasma fibrinogen concentration and platelet count, as well as to determine the overall survival of NSCLC patients with brain metastases. A total of 275 NSCLC patients with brain metastasis were enrolled into this study. Univariate analysis showed that high plasma fibrinogen concentration was associated with age?65 years (P = 0.011), smoking status (P = 0.009), intracranial symptoms (P = 0.022), clinical T category (P = 0.010), clinical N category (P = 0.003), increased partial thromboplastin time (P < 0.001), and platelet count (P < 0.001). Patients with low plasma fibrinogen concentration demonstrated longer overall survival compared with those with high plasma fibrinogen concentration (median, 17.3 months versus 11.1 months; P?0.001). A similar result was observed for platelet counts (median, 16.3 months versus 11.4 months; P = 0.004). Multivariate analysis showed that both plasma fibrinogen concentration and platelet count were independent prognostic factors for NSCLC with brain metastases (R2 = 1.698, P < 0.001 and R2 = 1.699, P < 0.001, respectively). Our results suggest that high plasma fibrinogen concentration and platelet count indicate poor prognosis for NSCLC patients with brain metastases. Thus, these two biomarkers might be independent prognostic predictors for this subgroup of NSCLC patients. PMID:23958057

Zhu, Jian-Fei; Cai, Ling; Zhang, Xue-Wen; Wen, Yin-Sheng; Su, Xiao-Dong; Rong, Tie-Hua; Zhang, Lan-Jun

2014-02-01

305

Contracting bubbles in Hele-Shaw cells with a power-law fluid  

NASA Astrophysics Data System (ADS)

The problem of bubble contraction in a Hele-Shaw cell is studied for the case in which the surrounding fluid is of power-law type. A small perturbation of the radially symmetric problem is first considered, focussing on the behaviour just before the bubble vanishes, it being found that for shear-thinning fluids the radially symmetric solution is stable, while for shear-thickening fluids the aspect ratio of the bubble boundary increases. The borderline (Newtonian) case considered previously is neutrally stable, the bubble boundary becoming elliptic in shape with the eccentricity of the ellipse depending on the initial data. Further light is shed on the bubble contraction problem by considering a long thin Hele-Shaw cell: for early times the leading-order behaviour is one-dimensional in this limit; however, as the bubble contracts its evolution is ultimately determined by the solution of a Wiener-Hopf problem, the transition between the long thin limit and the extinction limit in which the bubble vanishes being described by what is in effect a similarity solution of the second kind. This same solution describes the generic (slit-like) extinction behaviour for shear-thickening fluids, the interface profiles that generalize the ellipses that characterize the Newtonian case being constructed by the Wiener-Hopf calculation.

McCue, Scott W.; King, John R.

2011-02-01

306

Fluid Shear Stress Alters the Hemostatic Properties of Endothelial Outgrowth Cells  

PubMed Central

Surface endothelialization is an attractive means to improve the performance of small diameter vascular grafts. While endothelial outgrowth cells (EOCs) are considered a promising source of autologous endothelium, the ability of EOCs to modulate coagulation-related blood activities is not well understood. The goal of this study was to assess the role of arterial flow conditions on the thrombogenic phenotype of EOCs. EOCs derived from baboon peripheral blood, as well as mature arterial endothelial cells from baboons, were seeded onto adsorbed collagen, then exposed to physiologic levels of fluid shear stress. For important hemostatic pathways, cellular responses to shear stress were characterized at the gene and protein level and confirmed with a functional assay for activated protein C (APC) activity. For EOCs, fluid shear stress upregulated gene and protein expression of anticoagulant and platelet inhibitory factors, including thrombomodulin, tissue factor pathway inhibitor, and nitric oxide synthase 3 (eNOS). Fluid shear stress significantly altered the functional activity of EOCs by increasing APC levels. This study demonstrates that fluid shear stress is an important determinant of EOC hemostatic properties. Accordingly, manipulation of EOC phenotype by mechanical forces may be important for the development of thrombo-resistant surfaces on engineered vascular implants. PMID:21787250

Ensley, Ann E.; Nerem, Robert M.; Anderson, Deirdre E.J.; Hanson, Stephen R.

2012-01-01

307

Mass spectrometric based analysis, characterization and applications of circulating cell free DNA isolated from human body fluids  

Microsoft Academic Search

In the past decade, cell free DNA, or circulating cell free DNA, or cell free circulating DNA, isolated from body fluids such as plasma\\/serum\\/urine has emerged as an important tool for clinical diagnostics. The molecular biology of circulating cell free DNA is poorly understood but there is currently an increased effort to understand the origin, mechanism of its circulation, and

Vaneet K. Sharma; Paul Vouros; James Glick

2011-01-01

308

Inverse Relationship of Serum Hepcidin Levels with CD4 Cell Counts in HIV-Infected Patients Selected from an Indonesian Prospective Cohort Study  

PubMed Central

Background Distortion of iron homeostasis may contribute to the pathogenesis of human immunodeficiency virus (HIV) infection and tuberculosis (TB). We studied the association of the central iron-regulatory hormone hepcidin with the severity of HIV and the association between hepcidin and other markers of iron homeostasis with development of TB. Methods Three groups of patients were selected from a prospective cohort of HIV-infected subjects in Bandung, Indonesia. The first group consisted of HIV-infected patients who started TB treatment more than 30 days after cohort enrollment (cases). The second group consisted of HIV-infected patients who were matched for age, gender and CD4 cell count to the cases group (matched controls). The third group consisted of HIV-infected patients with CD4 cell counts above 200 cells/mm3 (unmatched controls). Iron parameters including hepcidin were compared using samples collected at cohort enrollment, and compared with recently published reference values for serum hepcidin. Results A total of 127 HIV-infected patients were included, 42 cases together with 42 matched controls and 43 unmatched controls. Patients with advanced HIV infection had elevated serum hepcidin and ferritin levels. Hepcidin levels correlated inversely with CD4 cells and hemoglobin. Cases had significantly higher hepcidin and ferritin concentrations at cohort enrollment compared to matched controls, but these differences were fully accounted for by the cases who started TB treatment between day 31 and 60 after enrollment. Hepcidin levels were not different in those with or without hepatitis C infection. Conclusion Iron metabolism is distorted in advanced HIV infection with CD4 cell counts correlating inversely with serum hepcidin levels. High serum hepcidin levels and hyperferritinemia were found in patients starting TB treatment shortly after cohort enrollment, suggesting that these parameters have a predictive value for development of manifest TB in HIV-infected patients. PMID:24244576

Wisaksana, Rudi; de Mast, Quirijn; Alisjahbana, Bachti; Jusuf, Hadi; Sudjana, Primal; Indrati, Agnes R.; Sumantri, Rachmat; Swinkels, Dorine; van Crevel, Reinout; van der Ven, Andre

2013-01-01

309

Increased Lymphocyte Infiltration in Rheumatoid Arthritis Is Correlated with an Increase in LTi-like Cells in Synovial Fluid.  

PubMed

In this study, we compared the immune cell populations in rheumatoid arthritis (RA) synovial fluid, which shows lymphoid tissue-like structure, with those in tonsils, which are normal secondary lymphoid tissues. Firstly, we found that CD4(-)CD11b(+) macrophages were the major population in RA synovial fluid and that B cells were the major population in tonsils. In addition, synovial fluid from patients with osteoarthritis, which is a degenerative joint disease, contained CD4(+)CD11b(+) monocytes as the major immune cell population. Secondly, we categorized three groups based on the proportion of macrophages found in RA synovial fluid: (1) the macrophage-high group, which contained more than 80% macrophages; (2) the macrophage-intermediate group, which contained between 40% and 80% macrophages; and (3) the macrophage-low group, which contained less than 40% macrophages. In the macrophage-low group, more lymphoid tissue inducer (LTi)-like cells were detected, and the expression of OX40L and TRANCE in these cells was higher than that in the other groups. In addition, in this group, the suppressive function of regulatory T cells was downregulated. Finally, CXCL13 expression was higher in RA synovial fluid than in tonsils, but CCL21 expression was comparable in synovial fluid from all groups and in tonsils. These data demonstrate that increased lymphocyte infiltration in RA synovial fluid is correlated with an increase in LTi-like cells and the elevation of the chemokine expression. PMID:24385942

Koo, Jihye; Kim, Soochan; Jung, Woong Jae; Lee, Ye Eun; Song, Gwan Gyu; Kim, Kyung-Su; Kim, Mi-Yeon

2013-12-01

310

Direct demonstration of tubular fluid flow sensing by macula densa cells  

PubMed Central

Macula densa (MD) cells in the cortical thick ascending limb (cTAL) detect variations in tubular fluid composition and transmit signals to the afferent arteriole (AA) that control glomerular filtration rate [tubuloglomerular feedback (TGF)]. Increases in tubular salt at the MD that normally parallel elevations in tubular fluid flow rate are well accepted as the trigger of TGF. The present study aimed to test whether MD cells can detect variations in tubular fluid flow rate per se. Calcium imaging of the in vitro microperfused isolated JGA-glomerulus complex dissected from mice was performed using fluo-4 and fluorescence microscopy. Increasing cTAL flow from 2 to 20 nl/min (80 mM [NaCl]) rapidly produced significant elevations in cytosolic Ca2+ concentration ([Ca2+]i) in AA smooth muscle cells [evidenced by changes in fluo-4 intensity (F); F/F0 = 1.45 ± 0.11] and AA vasoconstriction. Complete removal of the cTAL around the MD plaque and application of laminar flow through a perfusion pipette directly to the MD apical surface essentially produced the same results even when low (10 mM) or zero NaCl solutions were used. Acetylated ?-tubulin immunohistochemistry identified the presence of primary cilia in mouse MD cells. Under no flow conditions, bending MD cilia directly with a micropipette rapidly caused significant [Ca2+]i elevations in AA smooth muscle cells (fluo-4 F/F0: 1.60 ± 0.12) and vasoconstriction. P2 receptor blockade with suramin significantly reduced the flow-induced TGF, whereas scavenging superoxide with tempol did not. In conclusion, MD cells are equipped with a tubular flow-sensing mechanism that may contribute to MD cell function and TGF. PMID:20719981

Sipos, Arnold; Vargas, Sarah

2010-01-01

311

TGF-? mediates proinflammatory seminal fluid signaling in human cervical epithelial cells.  

PubMed

The cervix is central to the female genital tract immune response to pathogens and foreign male Ags introduced at coitus. Seminal fluid profoundly influences cervical immune function, inducing proinflammatory cytokine synthesis and leukocyte recruitment. In this study, human Ect1 cervical epithelial cells and primary cervical cells were used to investigate agents in human seminal plasma that induce a proinflammatory response. TGF-?1, TGF-?2, and TGF-?3 are abundant in seminal plasma, and Affymetrix microarray revealed that TGF-?3 elicits changes in Ect1 cell expression of several proinflammatory cytokine and chemokine genes, replicating principal aspects of the Ect1 response to seminal plasma. The differentially expressed genes included several induced in the physiological response of the cervix to seminal fluid in vivo. Notably, all three TGF-? isoforms showed comparable ability to induce Ect1 cell expression of mRNA and protein for GM-CSF and IL-6, and TGF-? induced a similar IL-6 and GM-CSF response in primary cervical epithelial cells. TGF-? neutralizing Abs, receptor antagonists, and signaling inhibitors ablated seminal plasma induction of GM-CSF and IL-6, but did not alter IL-8, CCL2 (MCP-1), CCL20 (MIP-3?), or IL-1? production. Several other cytokines present in seminal plasma did not elicit Ect1 cell responses. These data identify all three TGF-? isoforms as key agents in seminal plasma that signal induction of proinflammatory cytokine synthesis in cervical cells. Our findings suggest that TGF-? in the male partner's seminal fluid may influence cervical immune function after coitus in women, and potentially be a determinant of fertility, as well as defense from infection. PMID:22706080

Sharkey, David J; Macpherson, Anne M; Tremellen, Kelton P; Mottershead, David G; Gilchrist, Robert B; Robertson, Sarah A

2012-07-15

312

Effect of Allium cepa L. on Lipopolysaccharide-Stimulated Osteoclast Precursor Cell Viability, Count, and Morphology Using 4?,6-Diamidino-2-phenylindole-Staining  

PubMed Central

Allium cepa L. is known to possess numerous pharmacological properties. Our aim was to examine the in vitro effects of Allium cepa L. extract (AcE) on Porphyromonas gingivalis LPS and Escherichia coli LPS-stimulated osteoclast precursor cells to determine cell viability to other future cell-based assays. Osteoclast precursor cells (RAW 264.7) were stimulated by Pg LPS (1??g/mL) and E. coli LPS (1??g/mL) in the presence or absence of different concentrations of AcE (10–1000??g/mL) for 5 days at 37°C/5% CO2. Resazurin reduction and total protein content assays were used to detect cell viability. AcE did not affect cell viability. Resazurin reduction assay showed that AcE, at up to 1000??g/mL, did not significantly affect cell viability and cellular protein levels. Additionally a caspase 3/7 luminescence assay was used to disclose apoptosis and there was no difference in apoptotic activity between tested groups and control group. Fluorescence images stained by DAPI showed no alteration on the morphology and cell counts of LPS-stimulated osteoclast precursor cells with the use of AcE in all tested concentrations when compared to control. These findings suggest that Allium cepa L. extract could be used for in vitro studies on Porphyromonas gingivalis LPS and Escherichia coli LPS-stimulated osteoclast precursor cells. PMID:25221602

Oliveira, Tatiane; Figueiredo, Camila A.; Stavroullakis, Alexander; Da Silva Velozo, Eudes; Nogueira-Filho, Getulio

2014-01-01

313

Glutathione and GSH-dependent enzymes in bronchoalveolar lavage fluid cells in response to ozone  

SciTech Connect

The purpose of this study was to determine if in vivo ozone exposure results in elevations in the levels of glutathione and glutathione-dependent enzymes in cells derived from bronchoalveolar lavage fluid (BALF). Our hypothesis was that, as part of a defense mechanism against oxygen toxicity, such cells would have increased levels of glutathione (GSH) in response to an oxidant stress. Female F344/N rats were exposed to 0.8 ppm ozone, 6 hr/day, for 1, 3, or 7 days, after which cells were collected by lung lavage. The GSH and GSH-peroxidase activity per milligram of protein in the cellular fraction, both necessary for reducing cellular peroxides, were elevated after 3 days of ozone exposure. After 7 days of exposure, cellular GSH had returned to control values, but the activity of glutathione reductase, the enzyme that reduces oxidized glutathione to GSH, was increased. Extracellular GSH concentration and glutathione reductase activity in BALF were also increased after 7 days of exposure. The total glutathione equivalents (GSH and GSSG, both cellular and extracellular) in BALF increased throughout the 7-day exposure, with GSH increasing first in the cells, and then in the extracellular fluid. This study demonstrated that the glutathione anti-oxidant system of BALF cells is stimulated by exposure to ozone. This response may serve to protect cells from the toxic effects of oxidant stress.

Boehme, D.S.; Hotchkiss, J.A.; Henderson, R.F. (Inhalation Toxicology Research Institute, Lovelace Biomedical and Environmental Research Institute, Albuquerque, NM (United States))

1992-02-01

314

Amniotic Fluid-Derived Stem Cells for Cardiovascular Tissue Engineering Applications  

PubMed Central

Recent research has demonstrated that a population of stem cells can be isolated from amniotic fluid removed by amniocentesis that are broadly multipotent and nontumorogenic. These amniotic fluid-derived stem cells (AFSC) could potentially provide an autologous cell source for treatment of congenital defects identified during gestation, particularly cardiovascular defects. In this review, the various methods of isolating, sorting, and culturing AFSC are compared, along with techniques for inducing differentiation into cardiac myocytes and endothelial cells. Although research has not demonstrated complete and high-yield cardiac differentiation, AFSC have been shown to effectively differentiate into endothelial cells and can effectively support cardiac tissue. Additionally, several tissue engineering and regenerative therapeutic approaches for the use of these cells in heart patches, injection after myocardial infarction, heart valves, vascularized scaffolds, and blood vessels are summarized. These applications show great promise in the treatment of congenital cardiovascular defects, and further studies of isolation, culture, and differentiation of AFSC will help to develop their use for tissue engineering, regenerative medicine, and cardiovascular therapies. PMID:23350771

Petsche Connell, Jennifer; Camci-Unal, Gulden; Khademhosseini, Ali

2013-01-01

315

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

PubMed Central

We report how cell rheology measurements can be performed by monitoring the deformation of a cell in a microfluidic constriction, provided that friction and fluid leaks effects between the cell and the walls of the microchannels are correctly taken into account. Indeed, the mismatch between the rounded shapes of cells and the angular cross-section of standard microfluidic channels hampers efficient obstruction of the channel by an incoming cell. Moreover, friction forces between a cell and channels walls have never been characterized. Both effects impede a quantitative determination of forces experienced by cells in a constriction. Our study is based on a new microfluidic device composed of two successive constrictions, combined with optical interference microscopy measurements to characterize the contact zone between the cell and the walls of the channel. A cell squeezed in a first constriction obstructs most of the channel cross-section, which strongly limits leaks around cells. The rheological properties of the cell are subsequently probed during its entry in a second narrower constriction. The pressure force is determined from the pressure drop across the device, the cell velocity, and the width of the gutters formed between the cell and the corners of the channel. The additional friction force, which has never been analyzed for moving and constrained cells before, is found to involve both hydrodynamic lubrication and surface forces. This friction results in the existence of a threshold for moving the cells and leads to a non-linear behavior at low velocity. The friction force can nevertheless be assessed in the linear regime. Finally, an apparent viscosity of single cells can be estimated from a numerical prediction of the viscous dissipation induced by a small step in the channel. A preliminary application of our method yields an apparent loss modulus on the order of 100 Pa s for leukocytes THP-1 cells, in agreement with the literature data. PMID:24404016

Preira, Pascal; Valignat, Marie-Pierre; Bico, José; Théodoly, Olivier

2013-01-01

316

Actin-dependent fluid-phase endocytosis in inner cortex cells of maize root apices  

Microsoft Academic Search

The fluorescent dye Lucifer Yellow (LY) is a well- known and widely-used marker for fluid-phase endo- cytosis. In this paper, both light and electron micro- scopy revealed that LY was internalized into transition zone cells of the inner cortex of intact maize root apices. The internalized LY was localized within tubulo-vesicular compartments invaginating from the plasma membrane at actomyosin-enriched pit-fields

F. Baluska; J. Samaj; A. Hlavacka; J. Kendrick-Jones; D. Volkmann

2004-01-01

317

A standardized protocol for flow cytometric analysis of cells isolated from cerebrospinal fluid.  

PubMed

Flow cytometry (FC) is an useful tool for the analysis of subpopulations in complex cell suspensions. When applying this method to the cerebrospinal fluid (CSF), some characteristic properties of this cell type must be taken into consideration: there are only few cells which decay rapidly in their native medium and during centrifugation. One aim of the immunostaining procedure preceding flow cytometric analysis must be to minimize cell loss in order to get an undistorted picture of 'true' CSF cell populations. Consequently, morphological flow cytometric plots of high resolution are an indispensable precondition for reliable determination of subpopulations defined by monoclonal antibody (Mab) binding. We describe a standardized protocol for the flow cytometric examination of CSF cells which minimizes undesired cell loss. By the use of a 'quality control' the extent of cell loss could be monitored. Examples of morphological flow cytometric plots are given. The subsequent determination of Mab binding subpopulations is critical when fluorescence intensities of antigen positive and negative cells are non-disjunct. A statistical test was developed for these cases often seen when cell surface determinants are expressed at low levels only. PMID:7510788

Dux, R; Kindler-Röhrborn, A; Annas, M; Faustmann, P; Lennartz, K; Zimmermann, C W

1994-01-01

318

Effect of small parameters on the structure of the front formed by unstable viscous-fluid displacement from a Hele-Shaw Cell  

Microsoft Academic Search

The process of displacement of a viscous fluid from a Hele-Shaw cell consisting of two plates separated by a small gap is\\u000a investigated. The front formed when the fluid is displaced from the cell by another, lower-viscosity fluid is unstable. The\\u000a lower-viscosity fluid breaks through the layer of displaced fluid and forms channels called viscous fingers. As a result,\\u000a a

A. V. Zvyagin; O. E. Ivashnev; O. A. Logvinov

2007-01-01

319

Human Amniotic Fluid Mesenchymal Stem Cells in Combination with Hyperbaric Oxygen Augment Peripheral Nerve Regeneration  

Microsoft Academic Search

Purpose Attenuation of pro-inflammatory cytokines and associated inflammatory cell deposits rescues human amniotic fluid mesenchymal\\u000a stem cells (AFS) from apoptosis. Hyperbaric oxygen (HBO) suppressed stimulus-induced pro-inflammatory cytokine production\\u000a in blood-derived monocyte-macrophages. Herein, we evaluate the beneficial effect of hyperbaric oxygen on transplanted AFS\\u000a in a sciatic nerve injury model. Methods Peripheral nerve injury was produced in Sprague-Dawley rats by crushing

Hung-Chuan Pan; Chun-Shih Chin; Dar-Yu Yang; Shu-Peng Ho; Chung-Jung Chen; Shiaw-Min Hwang; Ming-Hong Chang; Fu-Chou Cheng

2009-01-01

320

Short Communication: Evaluation of the Overall Accuracy of the DeLaval Cell Counter for Somatic Cell Count in Ovine Milk: Effect of Soak Time in Diluted and Undiluted Milk Samples  

Microsoft Academic Search

This study evaluated the performance of the DeLaval cell counter (DCC) when analyzing ovine milk with dif- ferent soak times (defined as the permanence time of samples within the DCC cassette before starting the DCC counting procedure) in diluted and undiluted milk samples in 2 dairy sheep breeds. A total of 101 compos- ite ovine milk samples (50 from Assaf

C. Gonzalo; B. Linage; J. A. Carriedo; L. F. De La Fuente

2008-01-01

321

H Netten, I.T. Young, M. Prins, L.J. van Vliet, H. Tanke, J. Vrolijk, W. Sloos, Automation of dot counting in cell nuclei, in: Proc. 12th  

E-print Network

counting in cell nuclei, in: Proc. 12th IAPR International Conference on Pattern Recognition, Jerusalem of the normal two chromosomes. This particular aberration is associated with Down's syndrome. The labeling

van Vliet, Lucas J.

322

H Netten, I.T. Young, M. Prins, L.J. van Vliet, H. Tanke, J. Vrolijk, W. Sloos, Automation of dot counting in cell nuclei, in: Proc. 12 th IAPR International Conference on Pattern Recognition, Jerusalem, Israel, 1994, 8487.  

E-print Network

counting in cell nuclei, in: Proc. 12 th IAPR International Conference on Pattern Recognition, Jerusalem of the normal two chromosomes. This particular aberration is associated with Down's syndrome. The labeling

van Vliet, Lucas J.

323

Proteomic analysis of Staphylococcus aureus biofilm cells grown under physiologically relevant fluid shear stress conditions  

PubMed Central

Background The biofilm forming bacterium Staphylococcus aureus is responsible for maladies ranging from severe skin infection to major diseases such as bacteremia, endocarditis and osteomyelitis. A flow displacement system was used to grow S. aureus biofilms in four physiologically relevant fluid shear rates (50, 100, 500 and 1000 s-1) to identify proteins that are associated with biofilm. Results Global protein expressions from the membrane and cytosolic fractions of S. aureus biofilm cells grown under the above shear rate conditions are reported. Sixteen proteins in the membrane-enriched fraction and eight proteins in the cytosolic fraction showed significantly altered expression (p?fluid shear. These 24 proteins were identified using nano-LC-ESI-MS/MS. They were found to be associated with various metabolic functions such as glycolysis / TCA pathways, protein synthesis and stress tolerance. Increased fluid shear stress did not influence the expression of two important surface binding proteins: fibronectin-binding and collagen-binding proteins. Conclusions The reported data suggest that while the general metabolic function of the sessile bacteria is minimal under high fluid shear stress conditions, they seem to retain the binding capacity to initiate new infections. PMID:24855455

2014-01-01

324

Microbiological quality and somatic cell count in bulk milk of dromedary camels (Camelus dromedarius): descriptive statistics, correlations, and factors of variation.  

PubMed

The objectives of the present study were to monitor the microbiological quality and somatic cell count (SCC) of bulk tank milk at the world's first large-scale camel dairy farm for a 2-yr period, to compare the results of 2 methods for the enumeration of SCC, to evaluate correlation among milk quality indicators, and to determine the effect of specific factors (year, season, stage of lactation, and level of production) on milk quality indicators. The study was conducted from January 2008 to January 2010. Total viable count (TVC), coliform count (CC), California Mastitis Test (CMT) score, and SCC were determined from daily bulk milk samples. Somatic cell count was measured by using a direct microscopic method and with an automatic cell counter. In addition, production parameters [total daily milk production (TDM, kg), number of milking camels (NMC), average milk per camel (AMC, kg)] and stage of lactation (average postpartum days, PPD) were recorded for each test day. A strong correlation (r=0.33) was found between the 2 methods for SCC enumeration; however, values derived using the microscopic method were higher. The geometric means of SCC and TVC were 394×10(3) cells/mL and 5,157 cfu/mL during the observation period, respectively. Somatic cell count was >500×10(3) cells/mL on 14.6% (106/725) and TVC was >10×10(3) cfu/mL on 4.0% (30/742) of the test days. Both milk quality indicators had a distinct seasonal pattern. For log SCC, the mean was lowest in summer and highest in autumn. The seasonal pattern of log TVC was slightly different, with the lowest values being recorded during the spring. The monthly mean TVC pattern showed a clear difference between years. Coliform count was <10 cfu/mL in most of the samples (709/742, 95.6%). A positive correlation was found between log SCC and log TVC (r=0.32), between log SCC and CMT score (r=0.26), and between log TVC and CC in yr 1 (r=0.30). All production parameters and stage of lactation showed strong seasonal variation. Log SCC was negatively correlated with TDM (r=-0.35), AMC (r=-0.37), and NMC (r=-0.15) and positively correlated with PPD (r=0.40). Log TVC had a negative correlation with AMC (r=-0.40) but a positive correlation with NMC (r=0.32), TDM (r=0.16), and PPD (r=0.45). The linear mixed model with stepwise variable selection showed that the main sources of log SCC variation were PPD, TDM, PPD × season, and season. For log TVC, the same factors and year contributed to the variation. PMID:23849636

Nagy, P; Faye, B; Marko, O; Thomas, S; Wernery, U; Juhasz, J

2013-09-01

325

Human amniotic fluid stem cells suppress PBMC proliferation through IDO and IL-10-dependent pathways.  

PubMed

Human amniotic fluid stem cells (hAFSCs) can be readily isolated from human amniotic fluid and display multi-differentiation potential and immunomodulatory properties. The mechanism of hAFSCs immunoregulation has not been defined. Here, we explore the immunomodulatory effects of hAFSCs derived from human amniotic fluid and evaluate the role of IL-10 and the tryptophan-catabolizing enzyme indoleamine 2,3-dioxygenase (IDO) in mediating the immunosuppressive actions of hAFSCs. Flow cytometry showed that hAFSCs were positive for the mesenchymal stem cell markers CD29, CD44, CD105, HLA-ABC, and more than 84% of the hAFSCs were positive for SSEA-4, which is a typical marker of embryonic stem cell (ESCs), and negative for HLA-DR. The RT-PCR and immunostaining results revealed that the multipotent stem cells expressed OCT-4, Nanog, CD44, SOX2 and SSEA-1. In vitro differentiation assays demonstrated that hAFSCs underwent osteogenic differentiation. We examined the immunomodulatory function of hAFSCs using a co-culture system with phorbol 12-myristate 13-acetate (PMA) stimulated peripheral blood mononuclear cells (PBMCs). PBMC proliferation was suppressed by the hAFSCs in a dose-dependent manner. The inhibitory effect was caused by increased IL-10 and IDO induction after co-culture. Neutralizing the IL-10 activity or blocking the function of IDO partially abolished the immunosuppressive action of the hAFSCs. In conclusion, these results suggest that the hAFSCs possess immunomodulatory properties, and IL-10 and IDO are involved in immunosuppression by hAFSCs. PMID:24102581

Luo, Chengfeng; Jia, Wenwen; Wang, Kai; Chi, Fengli; Gu, Yanqiong; Yan, Xiaoling; Zou, Gang; Duan, Tao; Zhou, Qian

2014-01-01

326

New apparatus for direct counting of. beta. particles from two-dimensional gels and an application to changes in protein synthesis due to cell density  

SciTech Connect

A new method is described for scanning two-dimensional gels by the direct counting of ..beta.. particles instead of autoradiography. The methodology is described; results are compared with autoradiographic results; and data are presented demonstrating changed patterns of protein synthesis accompanying changes in cell density. The method is rapid and permits identification of differences in protein abundance of approximately 10% for a substantial fraction of the more prominent proteins. A modulation effect of more than 5 standard deviations, accompanying contact inhibition of cell growth, is shown to occur for an appreciable number of these proteins. The method promises to be applicable to a variety of biochemical and genetic experiments designed to delineate changes in protein synthesis accompanying changes in genome, molecular environment, history, and state of differentiation of the cell populations studied. 13 refs., 8 figs., 4 tabs.

Anderson, H.L.; Puck, T.T.; Shera, E.B.

1987-07-01

327

Natural evolution of CD4+ cell count in patients with CD4 >350 or >500? cells/mm3 at the time of diagnosis according to HIV-1 coreceptor tropism.  

PubMed

The HIV-1 coreceptor usage may play a critical role in AIDS pathogenesis and the X4-using viruses are considered to be more pathogenic than the R5-tropic viruses. These observations may influence the therapeutic decisions by asking for an earlier antiretroviral (ARV) treatment for the patients infected by the X4-tropic viruses compared with those infected by the R5-tropic viruses. The natural evolution of CD4+ cell count for 109 non-treated patients infected by the R5- or X4-tropic HIV-1 viruses with CD4+ >350 and >500 cells/mm(3) at time of diagnosis was compared until the initiation of an ARV regimen. The coreceptor usage was determined from the V3 env region sequence by Geno2Pheno (false positive rate 10%). A mixed linear regression model to analyse the CD4+ data with tropism as fixed effect in the model was used. Overall, 93 (85.3%) and 16 (14.7%) were infected by R5- and X4-tropic viruses, respectively. The median age, baseline CD4+ cell count, and viral load were 34 years (IQR: 30-42), 523 cells/mm(3) (IQR: 420-604), and 4.5 log(10) ?copies/ml (IQR: 3.9-5.0), respectively. There was no statistical difference in time to progression between the patients harboring R5- or X4-tropic viruses. The same results were observed for the sub-group of patients with CD4+ cell count >500 cells/mm(3). The virus tropism has no impact on the CD4+ cell count evolution in these HIV-1 patients diagnosed with CD4+ >350 or >500 cells/mm(3) suggesting that the tropism determination at time of diagnosis does not seem to be a useful tool to predict the clinical progression. PMID:23080487

Soulié, Cathia; Charpentier, Charlotte; Flandre, Philippe; Nino, Clément; Carcelain, Guislaine; Simon, Anne; Katlama, Christine; Landman, Roland; Brun-Vézinet, Françoise; Descamps, Diane; Calvez, Vincent; Marcelin, Anne-Geneviève

2012-12-01

328

Fluid flow releases fibroblast growth factor-2 from human aortic smooth muscle cells  

NASA Technical Reports Server (NTRS)

This study tested the hypothesis that fluid shear stress regulates the release of fibroblast growth factor (FGF)-2 from human aortic smooth muscle cells. FGF-2 is a potent mitogen that is involved in the response to vascular injury and is expressed in a wide variety of cell types. FGF-2 is found in the cytoplasm of cells and outside cells, where it associates with extracellular proteoglycans. To test the hypothesis that shear stress regulates FGF-2 release, cells were exposed to flow, and FGF-2 amounts were measured from the conditioned medium, pericellular fraction (extracted by heparin treatment), and cell lysate. Results from the present study show that after 15 minutes of shear stress at 25 dyne/cm(2) in a parallel-plate flow system, a small but significant fraction (17%) of the total FGF-2 was released from human aortic smooth muscle cells. FGF-2 levels in the circulating medium increased 10-fold over medium from static controls (P<0.01). A 50% increase in FGF-2 content versus control (P<0.01) was found in the pericellular fraction (extracted by heparin treatment). Furthermore, a significant decrease in FGF-2 was detected in the cell lysate, indicating that FGF-2 was released from inside the cell. Cell permeability studies with fluorescent dextran were performed to examine whether transient membrane disruption caused FGF-2 release. Flow cytometry detected a 50% increase in mean fluorescence of cells exposed to 25 dyne/cm(2) versus control cells. This indicates that the observed FGF-2 release from human aortic smooth muscle cells is likely due to transient membrane disruption on initiation of flow.

Rhoads, D. N.; Eskin, S. G.; McIntire, L. V.

2000-01-01

329

Mechanical interaction between cells and fluid for bone tissue engineering scaffold: Modulation of the interfacial shear stress  

E-print Network

/cell mechanical coupling was studied in a specific biological situation involving cells seeded in a bone scaffold place. The analytical model proposed in this study, while being a simplification of a fluid/cell mechanical interaction, brings complementary insights to numerical studies by analyzing the effect

Guerraoui, Rachid

330

Measurement of Inflammatory Mediators of Mast Cells and Eosinophils in Native Nasal Lavage Fluid in Nasal Polyposis  

Microsoft Academic Search

Background: Nasal polyposis (NP) often coexists with asthma, rhinitis and sinusitis. Polyp histology typically shows chronic, eosinophilic inflammation. The inflammatory cell infiltrate generally includes eosinophils, lymphocytes, plasma cells and mast cells. Objective: To gain insight into the natural history of NP, we analysed mediator levels and leukocyte values in nasal fluids and eosinophil cationic protein (ECP), total IgE levels and

Gabriele Di Lorenzo; Agata Drago; Maria Esposito Pellitteri; Giuseppina Candore; Alfredo Colombo; Francesco Gervasi; Maria Luisa Pacor; Francesco Purello D’Ambrosio; Calogero Caruso

2001-01-01

331

Fuel cell assembly unit for promoting fluid service and electrical conductivity  

DOEpatents

Fluid service and/or electrical conductivity for a fuel cell assembly is promoted. Open-faced flow channel(s) are formed in a flow field plate face, and extend in the flow field plate face between entry and exit fluid manifolds. A resilient gas diffusion layer is located between the flow field plate face and a membrane electrode assembly, fluidly serviced with the open-faced flow channel(s). The resilient gas diffusion layer is restrained against entering the open-faced flow channel(s) under a compressive force applied to the fuel cell assembly. In particular, a first side of a support member abuts the flow field plate face, and a second side of the support member abuts the resilient gas diffusion layer. The support member is formed with a plurality of openings extending between the first and second sides of the support member. In addition, a clamping pressure is maintained for an interface between the resilient gas diffusion layer and a portion of the membrane electrode assembly. Preferably, the support member is spikeless and/or substantially flat. Further, the support member is formed with an electrical path for conducting current between the resilient gas diffusion layer and position(s) on the flow field plate face.

Jones, Daniel O. (Glenville, NY)

1999-01-01

332

Long term persistent accumulation of CD8+ T cells in synovial fluid of rheumatoid arthritis  

PubMed Central

OBJECTIVE—To characterise the type and kinetics of T cell clones in synovial lesions of patients with rheumatoid arthritis (RA).?METHODS—Mononuclear cells from serial samples of synovial fluid (SF) and peripheral blood from nine RA patients were separated phenotypically using antibody coated magnetic beads. After mRNA preparation, reverse transcription-polymerase chain reaction (RT-PCR) was performed to amplify V-D(N)-J (that is, the third complementarity determining, CDR3) regions of their T cell receptor beta chain genes. This was followed by single strand conformation polymorphism (SSCP) analysis to detect the clonotypes of accumulating T cells. Amino acid sequences of the dominant clones were also determined.?RESULTS—Although peripheral T cells were heterogeneous, accumulation of oligoclonal T cells was detected in SF. The predominant accumulating clone was the CD8 subset, which was persistently present in serial samples obtained over almost one year of follow up. A proportion of these cells expressed CD25 or CD45RO, or both, suggesting they are `memory' clones.?CONCLUSION—The persistent presence of CD8+ T cell clones in RA joints indicates that they may be involved in the perpetuation of the chronic inflammatory process in RA joints.?? PMID:9389223

Masuko-Hongo, K.; Sekine, T.; Ueda, S.; Kobata, T.; Yamamoto, K.; Nishioka, K.; Kato, T.

1997-01-01

333

Effect of amniotic fluid on the in vitro culture of human corneal endothelial cells.  

PubMed

The present study was designed to evaluate the effects of human amniotic fluid (HAF) on the growth of human corneal endothelial cells (HCECs) and to establish an in vitro method for expanding HCECs. HCECs were cultured in DMEM-F12 supplemented with 20% fetal bovine serum (FBS). Confluent monolayer cultures were trypsinized and passaged using either FBS- or HAF-containing media. Cell proliferation and cell death ELISA assays were performed to determine the effect of HAF on cell growth and viability. The identity of the cells cultured in 20% HAF was determined using immunocytochemistry (ICC) and real-time reverse transcription polymerase chain reaction (RT-PCR) techniques to evaluate the expression of factors that are characteristic of HCECs, including Ki-67, Vimentin, Na+/K+-ATPase and ZO-1. HCEC primary cultures were successfully established using 20% HAF-containing medium, and these cultures demonstrated rapid cell proliferation according to the cell proliferation and death ELISA assay results. The ICC and real time RT-PCR results indicated that there was a higher expression of Na+/K+-ATPase and ZO-1 in the 20% HAF cell cultures compared with the control (20% FBS) (P < 0.05). The 20% HAF-containing medium exhibited a greater stimulatory effect on HCEC growth and could represent a potential enriched supplement for HCEC regeneration studies. PMID:24726921

Feizi, Sepehr; Soheili, Zahra-Soheila; Bagheri, Abouzar; Balagholi, Sahar; Mohammadian, Azam; Rezaei-Kanavi, Mozhgan; Ahmadieh, Hamid; Samiei, Shahram; Negahban, Kambiz

2014-05-01

334

Drug Transporters on Arachnoid Barrier Cells Contribute to the Blood-Cerebrospinal Fluid Barrier  

PubMed Central

The subarachnoid space, where cerebrospinal fluid (CSF) flows over the brain and spinal cord, is lined on one side by arachnoid barrier (AB) cells that form part of the blood-CSF barrier. However, despite the fact that drugs are administered into the CSF and CSF drug concentrations are used as a surrogate for brain drug concentration following systemic drug administration, the tight-junctioned AB cells have never been examined for whether they express drug transporters that would influence CSF and central nervous system drug disposition. Hence, we characterized drug transporter expression and function in AB cells. Immunohistochemical analysis showed P-glycoprotein (P-gp) and breast cancer resistance protein (BCRP) in mouse AB cells but not other meningeal tissue. The Gene Expression Nervous System Atlas (GENSAT) database and the Allen Mouse Brain Atlas confirmed these observations. Microarray analysis of mouse and human arachnoidal tissue revealed expression of many drug transporters and some drug-metabolizing enzymes. Immortalized mouse AB cells express functional P-gp on the apical (dura-facing) membrane and BCRP on both apical and basal (CSF-facing) membranes. Thus, like blood-brain barrier cells and choroid plexus cells, AB cells highly express drug transport proteins and likely contribute to the blood-CSF drug permeation barrier. PMID:23298861

Yasuda, Kazuto; Cline, Cynthia; Vogel, Peter; Onciu, Mihaela; Fatima, Soghra; Sorrentino, Brian P.; Thirumaran, Ranjit K.; Ekins, Sean; Urade, Yoshihiro; Fujimori, Ko

2013-01-01

335

Characteristics of silicone fluid as a pressure transmitting medium in diamond anvil cells  

NASA Astrophysics Data System (ADS)

The properties of a silicone fluid with initial viscosity of 1 cst as a pressure transmitting medium for diamond anvil cells have been determined by ruby R1 line broadening and R1-R2 separation measurements to 64 GPa at ambient temperature. By these criteria, the silicone fluid is as good a pressure medium as a 4:1 methanol:ethanol mixture at low pressures to about 20 GPa, and is better than the mixture at higher pressures. Although argon media are better than the silicone at pressures to 30 GPa, this silicone behaves as well as argon at higher pressures. Furthermore, the silicone is easier to load than argon and is almost chemically inert.

Shen, Yongrong; Kumar, Ravhi S.; Pravica, Michael; Nicol, Malcolm F.

2004-11-01

336

Amniotic Fluid-Derived Stem Cells Demonstrated Cardiogenic Potential in Indirect Co-culture with Human Cardiac Cells.  

PubMed

Amniotic fluid-derived stem cells (AFSC) have been shown to be broadly multipotent and non-tumorogenic. Previous studies of direct mixing of AFSC and neonatal rat ventricle myocytes indicated evidence of AFSC cardiogenesis. In this study, we examined human AFSC cardiogenic potential in indirect co-culture with human cardiac cells in conditions that eliminated the possibility of cell fusion. Human AFSC in contact with human cardiac cells showed expression of cardiac troponin T (cTnT) in immunohistochemistry, and no evidence of cell fusion was found through fluorescent in situ hybridization. When indirectly co-cultured with cardiac cells, human AFSC in contact with cardiac cells across a thin porous membrane showed a statistically significant increase in cTnT expression compared to non-contact conditions but lacked upregulation of calcium modulating proteins and did not have functional or morphological characteristics of mature cardiomyocytes. This suggests that contact is a necessary but not sufficient condition for AFSC cardiac differentiation in co-culture with cardiac cells. PMID:25266932

Gao, Yang; Connell, Jennifer Petsche; Wadhwa, Lalita; Ruano, Rodrigo; Jacot, Jeffrey G

2014-12-01

337

Adipogenic differentiation and EGFP gene transfection of amniotic fluid-derived stem cells from goat fetus at terminal gestational age.  

PubMed

The aims of this study were to determine whether stem cells could be isolated from amniotic fluid of goat fetus at terminal gestational age and to determine if these stem cells could differentiate into adipogenic cells and be transfected with a reporter gene, EGFP (enhanced green fluorescent protein). The stem cells were isolated from amniotic fluid of goat fetus at terminal gestational age, induced to differentiate into adipogenic cells in vitro and transfected with the EGFP gene using lipofection. Markers associated with undifferentiated AFS (amniotic fluid-derived stem) cells were tested by RT (reverse transcription)-PCR. The results demonstrated that AFS cells could be isolated from amniotic fluid of goat fetus at terminal gestational age and could differentiate into adipogenic cells. The EGFP gene was transfected into AFS cells successfully. EGFP gene transfection efficiency of the three groups of transgenic AFS cells were 26.0, 29.9 and 30.5%, respectively. Both transgenic and wild-type AFS cells could express Hes1 (hairy and enhancer of split 1), Oct4 (octamer-binding protein 4) and Nanog. PMID:21244367

He, Xiao-Ying; Zheng, Yue-Mao; Qiu, Shuang; Qi, Ying-Pei; Zhang, Yong

2011-08-01

338

Defect in recruiting effector memory CD8+ T-cells in malignant pleural effusions compared to normal pleural fluid  

PubMed Central

Background Malignant pleural effusions (MPE) are a common and fatal complication in cancers including lung or breast cancers, or malignant pleural mesothelioma (MPM). MPE animal models and immunotherapy trials in MPM patients previously suggested defects of the cellular immunity in MPE. However only few observational studies of the immune response were done in MPM patients, using questionable control groups (transudate…). Methods We compared T cell populations evaluated by flow cytometry from blood and pleural effusion of untreated patients with MPM (n?=?58), pleural metastasis of adenocarcinoma (n?=?30) or with benign pleural lesions associated with asbestos exposure (n?=?23). Blood and pleural fluid were also obtained from healthy subjects, providing normal values for T cell populations. Results Blood CD4+ or CD8+ T cells percentages were similar in all groups of patients or healthy subjects. Whereas pleural fluid from healthy controls contained mainly CD8+ T cells, benign or malignant pleural effusions included mainly CD4+ T cells. Effector memory T cells were the main T cell subpopulation in pleural fluid from healthy subjects. In contrast, there was a striking and selective recruitment of central memory CD4+ T cells in MPE, but not of effector cells CD8+ T cells or NK cells in the pleural fluid as one would expect in order to obtain an efficient immune response. Conclusions Comparing for the first time MPE to pleural fluid from healthy subjects, we found a local defect in recruiting effector CD8+ T cells, which may be involved in the escape of tumor cells from immune response. Further studies are needed to characterize which subtypes of effector CD8+ T cells are involved, opening prospects for cell therapy in MPE and MPM. PMID:23816056

2013-01-01

339

Human amniotic fluid stem cell injection therapy for urethral sphincter regeneration in an animal model  

PubMed Central

Background Stem cell injection therapies have been proposed to overcome the limited efficacy and adverse reactions of bulking agents. However, most have significant limitations, including painful procurement, requirement for anesthesia, donor site infection and a frequently low cell yield. Recently, human amniotic fluid stem cells (hAFSCs) have been proposed as an ideal cell therapy source. In this study, we investigated whether periurethral injection of hAFSCs can restore urethral sphincter competency in a mouse model. Methods Amniotic fluids were collected and harvested cells were analyzed for stem cell characteristics and in vitro myogenic differentiation potency. Mice underwent bilateral pudendal nerve transection to generate a stress urinary incontinence (SUI) model and received either periurethral injection of hAFSCs, periurethral injection of Plasma-Lyte (control group), or underwent a sham (normal control group). For in vivo cell tracking, cells were labeled with silica-coated magnetic nanoparticles containing rhodamine B isothiocyanate (MNPs@SiO2 (RITC)) and were injected into the urethral sphincter region (n = 9). Signals were detected by optical imaging. Leak point pressure and closing pressure were recorded serially after injection. Tumorigenicity of hAFSCs was evaluated by implanting hAFSCs into the subcapsular space of the kidney, followed two weeks later by retrieval and histologic analysis. Results Flow activated cell sorting showed that hAFSCs expressed mesenchymal stem cell (MSC) markers, but no hematopoietic stem cell markers. Induction of myogenic differentiation in the hAFSCs resulted in expression of PAX7 and MYOD at Day 3, and DYSTROPHIN at Day 7. The nanoparticle-labeled hAFSCs could be tracked in vivo with optical imaging for up to 10 days after injection. Four weeks after injection, the mean LPP and CP were significantly increased in the hAFSC-injected group compared with the control group. Nerve regeneration and neuromuscular junction formation of injected hAFSCs in vivo was confirmed with expression of neuronal markers and acetylcholine receptor. Injection of hAFSCs caused no in vivo host CD8 lymphocyte aggregation or tumor formation. Conclusions hAFSCs displayed MSC characteristics and could differentiate into cells of myogenic lineage. Periurethral injection of hAFSCs into an SUI animal model restored the urethral sphincter to apparently normal histology and function, in absence of immunogenicity and tumorigenicity. PMID:22906045

2012-01-01

340

Effects of Daflon 500 mg* on haemoconcentration and alterations of white blood cell count elicited by the upright position in anaesthetized dogs.  

PubMed

In the passive upright position, arterial and venous pressures in the human feet increase capillary pressure which leads to the filtration of fluid from the circulating plasma into the tissues of the feet. Loss of fluid concentrates both red cells and plasma so that the haematrocrit and plasma protein concentration of venous blood leaving the feet greatly exceed their mean values in the circulation. To study this phenomenon in animals, we used Beagle dogs in upright position. In blood of saphenous vein, red cells, haematocrit and plasma protein concentration have been studied. As in human (Moyses et al. Haemoconcentration and accumulation of white cells in the feet during venous stasis. Int J Microcirc Clin Exp 1987;5:311-20) red cells, haematocrit and plasma protein concentration increase in upright position. The increases in red cells, haematocrit and plasma protein concentration were higher and levels were greater after 2 hours when compared to the corresponding values after 1 hour. Daflon 500 mg, a micronized purified flavonoidic fraction, (200 mg/kg-1 per os) administered 20 minutes before upright position, significantly reduced these increases. This model might be a suitable model to test drugs interfering with venous stasis. PMID:8919260

Delbarre, B; Delbarre, G; Pillion, G; Calinon, F

1995-09-01

341

Cell-cell Interaction Underlies Formation of Fluid in the Male Reproductive Tract of the Rat  

PubMed Central

The epithelia lining the epididymides of many species consists of several cell types. We have provided evidence that the basal cells are essential to the integrated functions of the epithelium. Basal cells, but not principal cells, and other cells in the epididymis express TRPC3 and COX-1. We have isolated basal cells from intact rat epididymis using antibody-coated Dynabeads and subjected them to whole-cell patch-clamp measurement of nonselective cation channel activity, a feature of TRPC3 protein, and Fluo-3 fluorescence measurement of intracellular Ca2+ concentration. The results show that a nonselective cation current blockable by La3+ (0.1 mM), Gd3+ (0.1 mM), or SKF96365 (20 ?M) could be activated by lysylbradykinin (200 nM). In cells loaded with Fluo-3, addition of lysylbradykinin (100 nM) caused a sustained increase of intracellular Ca2+. This effect was blocked by Gd3+ (0.1 mM) or SKF96365 (20 ?M) and was not observed in Fluo-3–loaded principal cells. Stimulation of basal cell/principal cell cocultures with lysylbradykinin (200 nM) evoked in principal cells a current with CFTR-Cl? channel characteristics. Isolated principal cells in the absence of basal cells did not respond to lysylbradykinin but responded to PGE2 (100 nM) with activation of a CFTR-like current. Basal cells, but not principal cells, released prostaglandin E2 when stimulated with lysylbradykinin (100 nM). The release was blocked by SKF96365 (20 ?M) and BAPTA-AM (0.05 or 0.1 mM). Confluent cell monolayers harvested from a mixture of disaggregated principal cells and basal cells responded to lysylbradykinin (100 nM) and PGE2 (500 nM) with an increase in electrogenic anion secretion. The former response was dependent on prostaglandin synthesis as piroxicam blocked the response. However, cell cultures obtained from principal cells alone responded to PGE2 but not to bradykinin. These results support the notion that basal cells regulate principal cells through a Ca2+ and COX signaling pathway. PMID:15851503

Cheung, King-ho; Leung, George P.H.; Leung, Matthew C.T.; Shum, Winnie W.C.; Zhou, Wen-liang; Wong, Patrick Y.D.

2005-01-01

342

Death rates in HIV-positive antiretroviral-naive patients with CD4 count greater than 350 cells per microL in Europe and North America: a pooled cohort observational study  

PubMed Central

Background It is unclear whether antiretroviral (ART) naive HIV-positive individuals with high CD4 counts have a raised mortality risk compared with the general population, but this is relevant for considering earlier initiation of antiretroviral therapy. Methods Pooling data from 23 European and North American cohorts, we calculated country-, age-, sex-, and year-standardised mortality ratios (SMRs), stratifying by risk group. Included patients had at least one pre-ART CD4 count above 350 cells/mm3. The association between CD4 count and death rate was evaluated using Poisson regression methods. Findings Of 40,830 patients contributing 80,682 person-years of follow up with CD4 count above 350 cells/mm3, 419 (1.0%) died. The SMRs (95% confidence interval) were 1.30 (1.06-1.58) in homosexual men, and 2.94 (2.28-3.73) and 9.37 (8.13-10.75) in the heterosexual and IDU risk groups respectively. CD4 count above 500 cells/mm3 was associated with a lower death rate than 350-499 cells/mm3: adjusted rate ratios (95% confidence intervals) for 500-699 cells/mm3 and above 700 cells/mm3 were 0.77 (0.61-0.95) and 0.66 (0.52-0.85) respectively. Interpretation In HIV-infected ART-naive patients with high CD4 counts, death rates were raised compared with the general population. In homosexual men this was modest, suggesting that a proportion of the increased risk in other groups is due to confounding by other factors. Even in this high CD4 count range, lower CD4 count was associated with raised mortality. PMID:20638118

2011-01-01

343

Obesity Impairs Lymphatic Fluid Transport and Dendritic Cell Migration to Lymph Nodes  

PubMed Central

Introduction Obesity is a major cause of morbidity and mortality resulting in pathologic changes in virtually every organ system. Although the cardiovascular system has been a focus of intense study, the effects of obesity on the lymphatic system remain essentially unknown. The purpose of this study was to identify the pathologic consequences of diet induced obesity (DIO) on the lymphatic system. Methods Adult male wild-type or RAG C57B6-6J mice were fed a high fat (60%) or normal chow diet for 8–10 weeks followed by analysis of lymphatic transport capacity. In addition, we assessed migration of dendritic cells (DCs) to local lymph nodes, lymph node architecture, and lymph node cellular make up. Results High fat diet resulted in obesity in both wild-type and RAG mice and significantly impaired lymphatic fluid transport and lymph node uptake; interestingly, obese wild-type but not obese RAG mice had significantly impaired migration of DCs to the peripheral lymph nodes. Obesity also resulted in significant changes in the macro and microscopic anatomy of lymph nodes as reflected by a marked decrease in size of inguinal lymph nodes (3.4-fold), decreased number of lymph node lymphatics (1.6-fold), loss of follicular pattern of B cells, and dysregulation of CCL21 expression gradients. Finally, obesity resulted in a significant decrease in the number of lymph node T cells and increased number of B cells and macrophages. Conclusions Obesity has significant negative effects on lymphatic transport, DC cell migration, and lymph node architecture. Loss of T and B cell inflammatory reactions does not protect from impaired lymphatic fluid transport but preserves DC migration capacity. Future studies are needed to determine how the interplay between diet, obesity, and the lymphatic system modulate systemic complications of obesity. PMID:23950984

Weitman, Evan S.; Aschen, Seth Z.; Farias-Eisner, Gina; Albano, Nicholas; Cuzzone, Daniel A.; Ghanta, Swapna; Zampell, Jamie C.; Thorek, Daniel; Mehrara, Babak J.

2013-01-01

344

SOX9 as a Predictor for Neurogenesis Potentiality of Amniotic Fluid Stem Cells.  

PubMed

Preclinical studies of amniotic fluid-derived cell therapy have been successful in the research of neurodegenerative diseases, peripheral nerve injury, spinal cord injury, and brain ischemia. Transplantation of human amniotic fluid stem cells (AFSCs) into rat brain ventricles has shown improvement in symptoms of Parkinson's disease and also highlighted the minimal immune rejection risk of AFSCs, even between species. Although AFSCs appeared to be a promising resource for cell-based regenerative therapy, AFSCs contain a heterogeneous pool of distinct cell types, rendering each preparation of AFSCs unique. Identification of predictive markers for neuron-prone AFSCs is necessary before such stem cell-based therapeutics can become a reality. In an attempt to identify markers of AFSCs to predict their ability for neurogenesis, we performed a two-phase study. In the discovery phase of 23 AFSCs, we tested ZNF521/Zfp521, OCT6, SOX1, SOX2, SOX3, and SOX9 as predictive markers of AFSCs for neural differentiation. In the validation phase, the efficacy of these predictive markers was tested in independent sets of 18 AFSCs and 14 dental pulp stem cells (DPSCs). We found that high expression of SOX9 in AFSCs is associated with good neurogenetic ability, and these positive correlations were confirmed in independent sets of AFSCs and DPSCs. Furthermore, knockdown of SOX9 in AFSCs inhibited their neuronal differentiation. In conclusion, the discovery of SOX9 as a predictive marker for neuron-prone AFSCs could expedite the selection of useful clones for regenerative medicine, in particular, in neurological diseases and injuries. PMID:25154783

Wei, Pei-Cih; Chao, Angel; Peng, Hsiu-Huei; Chao, An-Shine; Chang, Yao-Lung; Chang, Shuenn-Dyh; Wang, Hsin-Shih; Chang, Yu-Jen; Tsai, Ming-Song; Sieber, Martin; Chen, Hua-Chien; Chen, Shu-Jen; Lee, Yun-Shien; Hwang, Shiaw-Min; Wang, Tzu-Hao

2014-10-01

345

Cardiac repair with injectable cell sheet fragments of human amniotic fluid stem cells in an immune-suppressed rat model.  

PubMed

Direct intramyocardial injection of the desired cell types in a dissociated form is a common route of cell transplantation for repair of damaged myocardium. However, following injection of dissociated cells, a massive loss of transplanted cells has been reported. In this study, human amniotic fluid stem cells (hAFSCs) were used as the cell source for the fabrication of cell sheet fragments, using a thermo-responsive methylcellulose hydrogel system. The fabricated hAFSC sheet fragments preserved the endogenous extracellular matrices (ECM) and retained their cell phenotype. Test samples were xenogenically transplanted into the peri-ischemic area of an immune-suppressed rat model at 1 week after myocardial infarction (MI) induction. There were four treatment groups (n>=10): sham; saline; dissociated hAFSCs; and hAFSC sheet fragments. The results obtained in the echocardiography revealed that the group treated with hAFSC sheet fragments had a superior heart function to those treated with saline or dissociated hAFSCs. Due to their inherent ECM, hAFSC sheet fragments had a better ability of cell retention and proliferation than dissociated hAFSCs upon transplantation to the host myocardium. Additionally, transplantation of hAFSC sheet fragments stimulated a significant increase in vascular density, consequently contributing towards improved wall thickness and a reduction in the infarct size, when compared with dissociated hAFSCs. Our histological findings and qPCR analyses suggest that the transplanted hAFSCs can be differentiated into cardiomyocyte-like cells and cells of endothelial lineages and modulate expression of multiple angiogenic cytokines and cardiac protective factor with the potential to promote neo-vascularization, which evidently contributed to the improvement of ventricular function. PMID:20621766

Yeh, Yi-Chun; Lee, Wen-Yu; Yu, Chu-Leng; Hwang, Shiaw-Min; Chung, Min-Fan; Hsu, Li-Wen; Chang, Yen; Lin, Wei-Wen; Tsai, Ming-Song; Wei, Hao-Ji; Sung, Hsing-Wen

2010-09-01

346

Flow Interactions with Cells and Tissues: Cardiovascular Flows and Fluid–Structure Interactions  

PubMed Central

Interactions between flow and biological cells and tissues are intrinsic to the circulatory, respiratory, digestive and genitourinary systems. In the circulatory system, an understanding of the complex interaction between the arterial wall (a living multi-component organ with anisotropic, nonlinear material properties) and blood (a shear-thinning fluid with 45% by volume consisting of red blood cells, platelets, and white blood cells) is vital to our understanding of the physiology of the human circulation and the etiology and development of arterial diseases, and to the design and development of prosthetic implants and tissue-engineered substitutes. Similarly, an understanding of the complex dynamics of flow past native human heart valves and the effect of that flow on the valvular tissue is necessary to elucidate the etiology of valvular diseases and in the design and development of valve replacements. In this paper we address the influence of biomechanical factors on the arterial circulation. The first part presents our current understanding of the impact of blood flow on the arterial wall at the cellular level and the relationship between flow-induced stresses and the etiology of atherosclerosis. The second part describes recent advances in the application of fluid–structure interaction analysis to arterial flows and the dynamics of heart valves. PMID:20336826

Friedman, Morton H.; Krams, Rob; Chandran, Krishnan B.

2010-01-01

347

Cutaneous presentation of chronic lymphocytic leukemia as unique extramedullar involvement in a patient with normal peripheral blood lymphocyte count (monoclonal B-cell lymphocytosis).  

PubMed

Skin infiltration by chronic lymphocytic leukemia (CLL) is very rare and almost all reported cases occur in advanced stage. We report a patient with no relevant past medical history who presented with cutaneous erythematous plaques. A punch biopsy showed typical CLL morphologic and immunophenotypic features. Subsequent studies revealed a normal lymphocyte count in peripheral blood, and there was no evidence of lymphadenopathy or organomegaly. Flow cytometry demonstrated a clonal B-cell population both in the bone marrow and peripheral blood (1.60 × 10(9)/l) with a CLL phenotype, but it did not fulfill required criteria for CLL diagnosis. Without cutaneous involvement, this case should be classified as monoclonal B-cell lymphocytosis. PMID:23639136

Tapia, Gustavo; Mate, José-Luis; Fuente, María-José; Navarro, José-Tomás; Fernández-Figueras, Maria-Teresa; Juncà, Jordi; Ferrándiz, Carlos; Ariza, Aurelio

2013-08-01

348

Human ovarian tumor ascites fluids rapidly and reversibly inhibit T cell receptor-induced NF-?B and NFAT signaling in tumor-associated T cells  

PubMed Central

Human memory T cells present in ovarian tumor ascites fluids fail to respond normally to stimulation via the T cell receptor (TCR). This immunosuppression is manifested by decreases in NF-?B and NFAT activation, IFN-? production, and cell proliferation in response to TCR stimulation with immobilized antibodies to CD3 and CD28. The anergy of the tumor-associated T cells (TATs) is mediated by soluble factors present in ovarian tumor ascites fluids. The non-responsiveness of the T cells is quickly reversed when the cells are assayed in the absence of the ascites fluid, and is rapidly reestablished when a cell-free ascites fluid is added back to the T cells. Based upon the observed normal phosphorylation patterns of the TCR proximal signaling molecules, the inhibition of NF-?B, and NFAT activation in response to TCR stimulation, as well as the ability of the diacylglycerol analog PMA and the ionophore ionomycin to bypass the ascites fluid-induced TCR signaling arrest, the site of the arrest in the activation cascade appears to be at or just upstream of PLC-?. An identical TCR signaling arrest pattern was observed when T cells derived from normal donor peripheral blood were incubated with either malignant or nonmalignant (cirrhotic) ascites fluids. The immunosuppressive activity of ascites fluids reported here suggests that soluble factors acting directly or indirectly upon T cells present within tumors contribute to the anergy that has previously been observed in T cells derived from malignant and nonmalignant inflammatory microenvironments. The soluble immunosuppressive factors represent potential therapeutic targets for ovarian cancer. PMID:23882159

Simpson-Abelson, Michelle R.; Loyall, Jenni L.; Lehman, Heather K.; Barnas, Jennifer L.; Minderman, Hans; O'Loughlin, Kieran L.; Wallace, Paul K.; George, Thaddeus C.; Peng, Peng; Kelleher, Raymond J.; Odunsi, Kunle; Bankert, Richard B.

2013-01-01

349

Regulation of bronchoalveolar lavage fluids cell function by the immunomodulatory agents from Cordyceps sinensis.  

PubMed

Cordyceps sinensis (C. sinensis) is one of the well known fungi used in traditional Chinese medicine for treatment asthma and bronchial and lung inflammation. In this study, effects of C. sinensis methanolic extracts on bronchoalveolar lavage fluids (BALF) cells proliferation, inflammatory cytokines production, and genes expression were evaluated. The proliferative response of BALF cells to lipopolysaccharide (LPS) was determined by the tritiated thymidine uptake method. The cell-free supernatants were harvested then tested for interlukin-1beta (IL-1beta), interlukin-6 (IL-6), tumor necrosis factor-alpha (TNF-alpha), interleukin-8 (IL-8), interleukin-10 (IL-10), interleukin-12 (IL-12), and interferon-gamma (IFN-gamma) by the enzyme immunoassay. The results indicated that the CS-19-22 fraction dose dependently suppressed BALF cells proliferation activated by LPS. The CS-19-22 fraction also reduced IL-1beta, IL-6, IL-8, IL-10 and TNF-alpha production in LPS activated BALF cell cultures. Furthermore, the IL-12 and IFN-gamma production in activated BALF cells were enhanced by CS-19-22 treatment. The CS-19-22 fraction did not affect IL-1beta, IL-6, TNF-alpha, and IL-8 mRNAs expression in BALF cells detected by reverse transcription-polymerase chain reaction (RT-PCR). By contrast, the CS-19-22 fraction increased IL-12 and IFN-gamma mRNAs expression and decreased IL-10 mRNA expression in the BALF cells activated with LPS. These results indicated the CS-19-22 fraction suppressed IL-1beta, IL-6, TNF-alpha, and IL-8 cytokines production in BALF cells through other than inhibition of mRNAs expression pathway. These results also demonstrate that the therapeutic activity of C. sinensis in Chinese medicine may be related to modulation of TH1 and TH2 cells functions in bronchial airway. PMID:11212870

Kuo, Y C; Tsai, W J; Wang, J Y; Chang, S C; Lin, C Y; Shiao, M S

2001-01-19

350

Absorbing Backside Anti-reflecting Layers for high contrast imaging in fluid cells  

E-print Network

The single Anti-Reflecting (AR) layer is a classical problem in optics. When all materials are pure dielectrics, the solution is the so-called lambda/4 layer. Here we examine the case of absorbing layers between non absorbing media. We find a solution for any layer absorption coefficient provided that the light goes from the higher towards the lower index medium, which characterizes backside layers. We describe these AR absorbing (ARA) layers through generalized index and thickness conditions. They are most often ultrathin, and have important applications for high contrast imaging in fluid cells.

Ausserré, Dominique; Amra, Claude; Zerrad, Myriam

2014-01-01

351

Membrane with internal passages to permit fluid flow and an electrochemical cell containing the same  

NASA Technical Reports Server (NTRS)

The invention provides an improved proton exchange membrane for use in electrochemical cells having internal passages parallel to the membrane surface, an apparatus and process for making the membrane, membrane and electrode assemblies fabricated using the membrane, and the application of the membrane and electrode assemblies to a variety of devices, both electrochemical and otherwise. The passages in the membrane extend from one edge of the membrane to another and allow fluid flow through the membrane and give access directly to the membrane for purposes of hydration.

Cisar, Alan J. (Inventor); Gonzalez-Martin, Anuncia (Inventor); Hitchens, G. Duncan (Inventor); Murphy, Oliver J. (Inventor)

1997-01-01

352

Multiple sclerosis: Brain-infiltrating CD8+ T cells persist as clonal expansions in the cerebrospinal fluid and blood  

PubMed Central

We surveyed the T cell receptor repertoire in three separate compartments (brain, cerebrospinal fluid, and blood) of two multiple sclerosis patients who initially had diagnostic brain biopsies to clarify their unusual clinical presentation but were subsequently confirmed to have typical multiple sclerosis. One of the brain biopsy specimens had been previously investigated by microdissection and single-cell PCR to determine the clonal composition of brain-infiltrating T cells at the single-cell level. Using complementarity-determining region 3 spectratyping, we identified several identical, expanded CD8+ (but not CD4+) T cell clones in all three compartments. Some of the expanded CD8+ T cells also occurred in sorted CD38+ blood cells, suggesting that they were activated. Strikingly, some of the brain-infiltrating CD8+ T cell clones persisted for >5 years in the cerebrospinal fluid and/or blood and may thus contribute to the progression of the disease. PMID:14983026

Skulina, Christian; Schmidt, Stephan; Dornmair, Klaus; Babbe, Holger; Roers, Axel; Rajewsky, Klaus; Wekerle, Hartmut; Hohlfeld, Reinhard; Goebels, Norbert

2004-01-01

353

Multipotent differentiation of the EGFP gene transgenic stem cells derived from amniotic fluid of goat at terminal gestational age.  

PubMed

We have isolated stem cells from amniotic fluid of goat at terminal gestational age and transferred the EGFP (enhanced green fluorescent protein) gene into the stem cells previously. The aim of this study was to determine whether the transgenic stem cells have the capability of multipotent differentiation. The transgenic stem cells were induced to differentiate into neurogenic, adipogenic, osteogenic and endothelial cells in vitro. Markers associated with AFS (amniotic fluid-derived stem) cells and the differentiated cells were tested by RT-PCR (reverse transcription-PCR). The results demonstrated that the transgenic AFS cells were capable of self-renewal, a defining property of stem cells. AFS cells were positive for the undifferentiated cell markers, Oct4, Nanog, Sox2 and Hes1, while following differentiation cells expressed markers for neurogenic cells such as astrocyte [GFAP (glial fibrillary acidic protein)] and NSE (neuron-specific enolase), adipogenic cells [LPL+ (lipoprotein lipase+)], osteogenic cells (osteocalcin+ and osteonectin+) and endothelium [CD34+ and eNOS+ (endothelial nitric oxide synthase)]. The results demonstrated that the EGFP gene transgenic AFS cells have the capability of multipotent differentiation, which means that the transgenic AFS cells may be useful in cell-transplantation studies in future. PMID:21605083

Zheng, Yue-Mao; Zheng, Yan-Ling; He, Xiao-Ying; He, Xiao-Ning; Zhao, Xue; Sai, Wu-Jia-Fu

2011-12-01

354

Computational fluid dynamics analysis in microbial fuel cells with different anode configurations.  

PubMed

A key criterion in microbial fuel cell (MFC) design is that the bio-electrochemical reaction between bacteria and the bulk solution should occur evenly on the electrode surface in order to improve electricity generation. However, experimental optimization of MFC design over a wide range of conditions is limited. Computational fluid dynamics (CFD) technology makes it possible to evaluate physicochemical phenomena such as fluid flows, mass transfer and chemical reaction, which can assist in system optimization. Twelve MFCs (M1-M12) with different internal structures were subjected to CFD analysis. The dead (DS) and working spaces (WS) of the anode compartment were calculated. The flow patterns of the anodic fluid varied according to the internal structures. The WS where the bio-electrochemical reaction can actually occur varied over the range of 0.14-0.57 m(2). Based on the above results, the power densities were estimated under the assumption that a monolayer biofilm was formed on the electrode. M11, with 18 rectangular-type internal structures, showed the largest WS of 0.57 m(2) and a theoretical maximum power density of 0.54 W/m(2). Although the optimization of the MFC configuration with only CFD analysis remains limited, the present study results are expected to provide fundamental data for MFC optimization. PMID:24718335

Kim, Jiyeon; Kim, Hongsuck; Kim, Byunggoon; Yu, Jaecheul

2014-01-01

355

Pleural fluid mononuclear cells (PFMCs) from tuberculous pleurisy can migrate in vitro in response to CXCL10.  

PubMed

We recently reported that pleural fluid mononuclear cells (PFMCs) from tuberculous pleurisy stimulated with Bacillus Calmette-Guérin (BCG) or TB antigens produced high levels of cytokines. However, it was still unclear what mechanism of the PFMCs used to migrate into the pleural fluids in TB infection. In the present study, we found that CD3(+)CD4(+) and CD3(+)CD8(+) T cells from PFMCs expressed significantly high levels of CXCR3 compared to PBMCs. In addition, the levels of CXCL10 (the ligand for CXCR3) in pleural fluids were significantly higher than those in normal serum and cancerous fluids. After stimulation with BCG, PFMCs produced high levels of CXCL10. Importantly, the synthesis of CXCL10 was mainly dependent on the BCG-induced production of IFNs, because the neutralization of endogenous IFN-? or IFN-? with mAbs significantly reduced the production of CXCL10 from BCG-stimulated PFMCs. In addition, the tubercular pleural fluid (TBPF) or exogenous CXCL10 induced the migration of PFMCs, indicating that IFN-? or IFN-? modulated the immune response through the expression of CXCL10 to aid the recruitment and selective homing of activated/effector cells to the site of Mycobacterium tuberculosis (M.tb) infection. Taken together, the levels of CXCL10 in pleural fluids were high and BCG-stimulated PFMCs expressed high levels of CXCL10, and CXCL10 induced the migration of PFMCs into the pleural fluids in TB infection. PMID:24406079

Wu, Changyou; Ma, Jiangjun; Xu, Yanquan; Zhang, Xianlan; Lao, Suihua; Yang, Binyan

2014-03-01

356

Receptor-independent fluid-phase pinocytosis mechanisms for induction of foam cell formation with native LDL particles  

PubMed Central

Purpose of review Because early findings indicated that native low density lipoprotein (LDL) did not substantially increase macrophage cholesterol content during in vitro incubations, investigators presumed that LDL must be modified in some way to trigger its uptake by the macrophage. The purpose of this review is to discuss recent findings showing that native unmodified LDL can induce massive macrophage cholesterol accumulation mimicking macrophage foam cell formation that occurs within atherosclerotic plaques. Recent findings Macrophages that show high rates of fluid-phase pinocytosis also show similar high rates of uptake of native unmodified LDL through non-receptor mediated uptake within both macropinosomes and micropinosomes. Non-saturable fluid-phase uptake of LDL by macrophages converts the macrophages into foam cells. Different macrophage phenotypes demonstrate either constitutive fluid-phase pinocytosis or inducible fluid-phase pinocytosis. Fluid-phase pinocytosis has been demonstrated by macrophages within mouse atherosclerotic plaques indicating that this pathway contributes to plaque macrophage cholesterol accumulation. Summary Contrary to what has been believed previously, macrophages can take up large amounts of native unmodified LDL by receptor-independent, fluid-phase pinocytosis converting these macrophages into foam cells. Thus, targeting macrophage fluid-phase pinocytosis should be considered when investigating strategies to limit macrophage cholesterol accumulation in atherosclerotic plaques. PMID:21881499

Kruth, Howard S.

2014-01-01

357

Spreading of acute myeloid leukemia cells by trafficking along the peripheral outflow pathway of cerebrospinal fluid.  

PubMed

Acute myeloid leukemia (AML) can affect not only bone marrow (BM) and peripheral blood (PB), but also the compartment of cerebrospinal fluid (CSF). Besides standard chemotherapy, specific and non-specific immunotherapy has been employed synergistically to treat AML patients. Here we report on a patient who received standard chemotherapy, unspecific immunotherapy with interleukin-2, as well as later specific CD8+ T-cell stimulation by RHAMM-R3 peptide vaccination. The patient maintained a complete remission in BM and PB, while he developed recurrent relapses in the CSF. Moreover, the patient developed a chloroma in the vicinity of neuronal sheaths during hematological CR, but high leukemia cell numbers within the CSF spaces over a long time period. This rare observation demonstrates several aspects. There is a previously unknown site of leukemia cell distribution, namely the peripheral cerebrospinal outflow pathway (PCOP). This demonstrates the ineffective therapy of this previously unknown mechanism of leukemia cell spread. The hypothesis that the PCOP is a site of physiological CSF-T-cell trafficking, and the lumen of the PCOP should be considered as an extension of the subarachnoidal spaces without closed anatomical borders is supported by this observation. The CSF spaces, but possibly specifically the PCOP, may represent a previously unknown survival niche of tumor cells within intrathecal spaces. PMID:21737662

Schmitt, Michael; Neubauer, Andreas; Greiner, Jochen; Xu, Xun; Barth, Thomas F E; Bechter, Karl

2011-06-01

358

Modeling neurogenesis impairment in Down syndrome with induced pluripotent stem cells from Trisomy 21 amniotic fluid cells.  

PubMed

Down syndrome (DS), or Trisomy 21 (T21) syndrome, one of the most common chromosomal abnormalities, is caused by an extra duplication of chromosome 21. In studies of neuron development, experimental models based on human cells are considered to be the most desired and accurate for basic research. The generation of diseased induced pluripotetn stem (iPS) cell is a critical step in understanding the developmental stages of complex neuronal diseases. Here, we generated human DS iPS cell lines from second trimester amniotic fluid (AF) cells with T21 by co-expressing Yamanaka factors through lentiviral delivery and subsequently differentiated them into neuronal progenitor cells (NPCs) for further analyses. T21 AF-iPS cells were characterized for the expression of pluripotent markers and for their ability to differentiate into all three germ layers by forming embryoid bodies in vitro and teratomas in vivo. The T21 AF-iPS cells maintained their unique pattern of chromosomal karyotypes: three pairs of chromosome 21. The level of amyloid precursor protein was significantly increased in NPCs derived from T21 AF-iPS cells compared with NPCs from normal AF-iPS cells. The expression levels of miR-155 and miR-802 in T21 AF-iPS-NPCs were highly elevated in the presence of low expression of MeCP2. We observed that T21 iPS-NPCs generated fewer neurons compared with controls. T21 iPS-NPCs exhibit developmental defects during neurogenesis. Our findings suggest that T21 AF-iPS cells serve as a good source to further elucidate the impairment neurogenesis of DS and the onset of Alzheimer's disease. PMID:23041301

Lu, Huai-En; Yang, Yao-Chen; Chen, Sheng-Mei; Su, Hong-Lin; Huang, Pai-Cheng; Tsai, Ming-Song; Wang, Tzu-Hao; Tseng, Ching-Ping; Hwang, Shiaw-Min

2013-02-15

359

Avian leucocyte counting using the hemocytometer  

USGS Publications Warehouse

Automated methods for counting leucocytes in avian blood are not available because of the presence of nucleated erythrocytes and thrombocytes. Therefore, total white blood cell counts are performed by hand using a hemocytometer. The Natt and Herrick and the Unopette methods are the most common stain and diluent preparations for this procedure. Replicate hemocytometer counts using these two methods were performed on blood from four birds of different species. Cells present in each square of the hemocytometer were counted. Counting cells in the corner, side, or center hemocytometer squares produced statistically equivalent results; counting four squares per chamber provided a result similar to that obtained by counting nine squares; and the Unopette method was more precise for hemocytometer counting than was the Natt and Herrick method. The Unopette method is easier to learn and perform but is an indirect process, utilizing the differential count from a stained smear. The Natt and Herrick method is a direct total count, but cell identification is more difficult.

Dein, F.J.; Wilson, A.; Fischer, D.; Langenberg, P.

1994-01-01

360

Dynamical analysis of erythrocytes under the assumption of cross-spectral coherence between blood cell counts and the Dst  

E-print Network

Dynamical analysis of erythrocytes under the assumption of cross-spectral coherence between blood type of blood cell are expected to reflect the intrinsic dynamics of the hematologic system and its for the transport of oxygen, coagulation and the immune response respectively. In the healthy subject, blood cell

Dasso, Sergio

361

Tagging Endogenous Loci for Live-Cell Fluorescence Imaging and Molecule Counting Using ZFNs, TALENs, and Cas9.  

PubMed

The programmable ZFN, TALEN, and Cas9 nucleases allow genome editing of any cell line or organism. In this chapter, we describe methods to create gene fusions at endogenous loci in mammalian cells to express fluorescent fusions of proteins of interest at endogenous levels. The donor DNA, which includes the sequence encoding a fluorescent protein, is provided to the cell to repair a double-strand break induced by a nuclease. The engineered donor sequence is integrated by homology-directed repair into the genome in frame with the coding region of the gene of interest, resulting in expression of a fusion protein at physiological levels. We further describe techniques to study protein dynamics and numbers using the genome-edited cell lines. In contrast to cell lines stably overexpressing fusion proteins from modified cDNAs, genes encoding fluorescent proteins are targeted to the endogenous genetic locus, avoiding perturbation of alternative splicing and expression levels. PMID:25398339

Dambournet, D; Hong, S H; Grassart, A; Drubin, D G

2014-01-01

362

Roles of cell confluency and fluid shear in 3-dimensional intracellular forces in endothelial cells  

PubMed Central

We use a novel 3D inter-/intracellular force microscopy technique based on 3D traction force microscopy to measure the cell–cell junctional and intracellular tensions in subconfluent and confluent vascular endothelial cell (EC) monolayers under static and shear flow conditions. We found that z-direction cell–cell junctional tensions are higher in confluent EC monolayers than those in subconfluent ECs, which cannot be revealed in the previous 2D methods. Under static conditions, subconfluent cells are under spatially non-uniform tensions, whereas cells in confluent monolayers are under uniform tensions. The shear modulations of EC cytoskeletal remodeling, extracellular matrix (ECM) adhesions, and cell–cell junctions lead to significant changes in intracellular tensions. When a confluent monolayer is subjected to flow shear stresses with a high forward component comparable to that seen in the straight part of the arterial system, the intracellular and junction tensions preferentially increase along the flow direction over time, which may be related to the relocation of adherens junction proteins. The increases in intracellular tensions are shown to be a result of chemo-mechanical responses of the ECs under flow shear rather than a direct result of mechanical loading. In contrast, the intracellular tensions do not show a preferential orientation under oscillatory flow with a very low mean shear. These differences in the directionality and magnitude of intracellular tensions may modulate translation and transcription of ECs under different flow patterns, thus affecting their susceptibility for atherogenesis. PMID:22665785

Hur, Sung Sik; del Alamo, Juan C.; Park, Joon Seok; Li, Yi-Shuan; Nguyen, Hong A.; Teng, Dayu; Wang, Kuei-Chun; Flores, Leona; Alonso-Latorre, Baldomero; Lasheras, Juan C.; Chien, Shu

2012-01-01

363

The use of human amniotic fluid mesenchymal stem cells as the feeder layer to establish human embryonic stem cell lines.  

PubMed

Human embryonic stem cells (hESCs) are pluripotent cells that have the potential to differentiate into the three germ layers and possibly all tissues of the human body. To fulfil the clinical potentials for cell-based therapy, banks of hESC lines that express different combinations of the major histocompatibility genes should be established, preferably without exposing such cells to animal cells and proteins. In this study, we tested human amniotic fluid mesenchymal stem cells (AFMSCs) as feeder cells to support the growth of hESCs. Our results indicated that mitomycin-treated AFMSCs were able to support the newly established hESC lines CGLK-1 and CGLK-2. The hESC colonies cultured on AFMSCs expressed alkaline phosphatase (ALK-P), SSEA-4, TRA-1-60, TRA-1-81, Oct-4, Nanog and Sox-2, which are markers for undifferentiated hESCs. Chromosomal analyses of both hESC lines, CGLK-1 and CGLK-2, which were cultured on AFMSC feeders for 22 and 14 passages, respectively, were confirmed to be normal karyotypes (46, XX). The ability of AFMSCs as feeder cells to maintain the undifferentiated growth and pluripotency of hESCs was confirmed by in vivo formation of teratomas derived on AFMSC hESCs in severe combined immune-compromised mice. The use of AFMSCs for feeder cells to culture hESCs has several advantages, in that AFMSCs are not tumourigenic and can be expanded extensively with a short doubling time. Copyright © 2013 John Wiley & Sons, Ltd. PMID:23460275

Soong, Yung-Kwei; Huang, Shang-Yu; Yeh, Chiu-Hsiang; Wang, Tzu-Hao; Chang, Kuo-Hsuan; Cheng, Po-Jen; Shaw, S W Steven

2013-03-01

364

Micro Flow Cytometry Chip Device Integrated with Tunable Microlens for Circulating Tumor Cells Detection and Counting Applications  

NASA Astrophysics Data System (ADS)

This paper reports a micromachine-based micro flow cytometry chip device integrated with a tunable liquid-filled microlens structure for optical detection, in which is capable of fluorescence detection for circulating tumor cells (CTCs) detection applications. By utilizing two sets of micro-piezoelectric pump device, cell samples can be aligned and focused to flow through the sensing area, and enhanced the accuracy and sensitivity of samples detection. The integrated chip device can be used to detect tumor cells labelled with specific fluorescent dyes successfully, and the optical signals can be enhanced accordingly by utilizing the tunable liquid-filled microlens within specific liquid pressure.

Hsiung, Suz-Kai; Lin, Shiu-Ru; Lin, Che-Hsin

2010-06-01

365

Rapid Separation and Concentration of Food-Borne Pathogens in Food Samples Prior to Quantification by Viable-Cell Counting and Real-Time PCR?  

PubMed Central

Buoyant density gradient centrifugation has been used to separate bacteria from complex food matrices, as well as to remove compounds that inhibit rapid detection methods, such as PCR, and to prevent false-positive results due to DNA originating from dead cells. Applying a principle of buoyant density gradient centrifugation, we developed a method for rapid separation and concentration following filtration and low- and high-speed centrifugation, as well as flotation and sedimentation buoyant density centrifugation, for 12 food-borne pathogens (Salmonella enterica, Escherichia coli, Yersinia enterocolitica, Campylobacter jejuni, Vibrio cholerae O139, Vibrio parahaemolyticus O3K6, Vibrio vulnificus, Providencia alcalifaciens, Aeromonas hydrophila, Bacillus cereus, Staphylococcus aureus, and Clostridium perfringens) in 13 different food homogenates. This method can be used prior to real-time quantitative PCR (RTi-qPCR) and viable-cell counting. Using this combined method, the target organisms in the food samples theoretically could be concentrated 250-fold and detected at cell concentrations as low as 101 to 103 CFU/g using the RTi-qPCR assay, and amounts as small as 100 to 101 CFU/g could be isolated using plate counting. The combined separation and concentration methods and RTi-qPCR confirmed within 3 h the presence of 101 to 102 CFU/g of Salmonella and C. jejuni directly in naturally contaminated chicken and the presence of S. aureus directly in remaining food items in a poisoning outbreak. These results illustrated the feasibility of using these assays for rapid inspection of bacterial food contamination during a real-world outbreak. PMID:17056684

Fukushima, Hiroshi; Katsube, Kazunori; Hata, Yukiko; Kishi, Ryoko; Fujiwara, Satomi

2007-01-01

366

Levels of CD105+ cells increase and cell proliferation decreases during S-phase arrest of amniotic fluid cells in long-term culture  

PubMed Central

The present study aimed to improve the characterization of amniotic fluid cells (AFCs) in order to optimize their use in chromosomal prenatal diagnosis and as seed or stem cells for tissue engineering. The AFCs used in the current study were obtained from three females in their second trimester of pregnancy. The cells were cultured independently and characterized by cell morphology, cell markers, cell cycle distribution and chromosome Giemsa banding in an early- and late-passage. The AFCs remained homogeneous in culture and expressed mesenchymal markers, but not endothelial markers along the culture process. In addition, compared with the early-passage cells, the late-passage cells exhibit an increase in CD105 expression, a decrease in cell division and a delay in the cell cycle, and a number of cells underwent cell cycle arrest. However, the cells retained a normal karyotype. Therefore, the current study characterized AFCs in a clinical culture and confirmed that AFCs are mesenchymal precursors. The results obtained may be useful for the application of AFCs in prenatal diagnosis.

WANG, DING; CHEN, RUI; ZHONG, XUAN; FAN, YONG; LAI, WEIQIANG; SUN, XIAOFANG

2014-01-01

367

Three-Dimensional Computational Fluid Dynamics Modeling of Solid Oxide Electrolysis Cells and Stacks  

SciTech Connect

A three-dimensional computational fluid dynamics (CFD) electrochemical model has been created for detailed analysis of a high-temperature electrolysis stack (solid oxide fuel cells operated as electrolyzers). Inlet and outlet plenum flow distributions are discussed. Maldistribution of plena flow show deviations in per-cell operating conditions due to non-uniformity of species concentrations. Models have also been created to simulate experimental conditions and for code validation. Comparisons between model predictions and experimental results are discussed. Mass, momentum, energy, and species conservation and transport are provided via the core features of the commercial CFD code FLUENT. A solid-oxide fuel cell (SOFC) model adds the electrochemical reactions and loss mechanisms and computation of the electric field throughout the cell. The FLUENT SOFC user-defined subroutine was modified for this work to allow for operation in the electrolysis mode. Model results provide detailed profiles of temperature, Nernst potential, operating potential, activation over-potential, anode-side gas composition, cathode-side gas composition, current density and hydrogen production over a range of stack operating conditions. Variations in flow distribution, and species concentration are discussed. End effects of flow and per-cell voltage are also considered. Predicted mean outlet hydrogen and steam concentrations vary linearly with current density, as expected. Contour plots of local electrolyte temperature, current density, and Nernst potential indicate the effects of heat transfer, reaction cooling/heating, and change in local gas composition.

Grant Hawkes; James O'Brien; Carl Stoots; Stephen Herring

2008-07-01

368

Fluid Shear Stress-Induced JNK Activity Leads to Actin Remodeling for Cell Alignment  

PubMed Central

Fluid shear stress (FSS) exerted on endothelial cell surfaces induces actin cytoskeleton remodeling through mechanotransduction. This study was designed to determine whether FSS activates Jun N-terminal kinase (JNK), to examine the spatial and temporal distribution of active JNK relative to the actin cytoskeleton in endothelial cells exposed to different FSS conditions, and to evaluate the effects of active JNK on actin realignment. Exposure to 15 and 20 dyn/cm2 FSS induced higher activity levels of JNK than the lower 2 and 4 dyn/cm2 flow conditions. At the higher FSS treatments, JNK activity increased with increasing exposure time, peaking 30 minutes after flow onset with an 8-fold activity increase compared to cells in static culture. FSS-induced phospho-JNK co-localized with actin filaments at cell peripheries, as well as with stress fibers. Pharmacologically blocking JNK activity altered FSS-induced actin structure and distribution as a response to FSS. Our results indicate that FSS-induced actin remodeling occurs in three phases, and that JNK plays a role in at least one, suggesting that this kinase activity is involved in mechanotransduction from the apical surface to the actin cytoskeleton in endothelial cells. PMID:20626006

Mengistu, Meron; Brotzman, Hannah; Ghadiali, Samir; Lowe-Krentz, Linda

2012-01-01

369

Dendritic cells in synovial fluid of chronic inflammatory arthritis lack CD80 surface expression.  

PubMed Central

Dendritic cells (DC) act as potent primary antigen-presenting cells in many immune responses and therefore may have a role in the initiation and perpetuation of the synovial inflammation in chronic inflammatory arthritis. To examine their function, it is important to isolate fresh DC from arthritic joints without aberrant activation. We have developed a technique using minimal cell manipulation to isolate DC from the synovial fluid of chronic arthritic patients. Using this method, DC were shown to be potent allostimulatory cells, with 63-90% of cells lacking lineage-specific markers (lin-), but positive for MHC class II molecules. Two morphologically distinct populations of these cells were identified in 10 out of 13 DC preparations. Both populations expressed CD40, intercellular adhesion molecule-1 (ICAM-1), ICAM-2, ICAM-3 and leucocyte function associated antigen-3 (LFA-3), but the predominant population, which was larger and more typical of cultured blood DC, had a higher density of these antigens compared with the minor population, which were smaller and morphologically similar to lymphocytes. Two new MoAbs which label activated human blood DC, HB15 (CD83) and CMRF-44, were tested. CD83 labelled very weakly or not at all, whereas CMRF-44 was positive on the larger cells only. Likewise, the costimulator molecule, B7/BB1 (CD80), was not detected on the surface of either synovial lin- cell population, reverse transcriptase polymerase chain reaction (RT-PCR) showed little or no CD80 mRNA, and no binding of the CTLA-4Ig fusion protein was found. These results suggest that synovial DC are not, despite the inflammatory environment, in a fully activated state. Images Fig. 2 Fig. 6 PMID:7535211

Summers, K L; Daniel, P B; O'Donnell, J L; Hart, D N

1995-01-01

370

Prevention of Infection in Patients With Hematologic Cancer and Persistent Fever Caused by a Low White Blood Cell Count  

ClinicalTrials.gov

Bone Marrow Suppression; Fever, Sweats, and Hot Flashes; Infection; Leukemia; Lymphoma; Multiple Myeloma and Plasma Cell Neoplasm; Myelodysplastic Syndromes; Unspecified Adult Solid Tumor, Protocol Specific; Unspecified Childhood Solid Tumor, Protocol Specific

2012-09-20

371

Native and rearranged ALK copy number and rearranged cell count in NSCLC: Implications for ALK inhibitor therapy  

PubMed Central

Background Anaplastic Lymphoma Kinase positive (ALK+) non-small cell lung cancer (NSCLC) responds to ALK inhibitors. Clinically, ? 15% cells showing rearrangements by break-apart FISH classify tumors as positive. Increases in native and rearranged ALK copy number also occur. Methods 1426 NSCLC clinical specimens (174 ALK+ and 1252 ALK negative), and 24 ALK negative NSCLC cell lines were investigated. ALK copy number and genomic status were assessed by FISH. Results Clinical specimens with 0–9%, 10–15%, 16–30%, 31–50% and >50% of ALK+ cells were found in 79.3%, 8.5%, 1.4%, 2.7% and 8.1% of cases, respectively. Increased native ALK copy number (?3 copies/cell in ?40% cells) was detected in 19% of ALK+ and 62% of ALK negative tumors. In ALK negative tumors, abundant focal amplification of native ALK was rare (0.8%). Other atypical patterns occurred in ~6% of tumors. Mean native ALK copy number ranged from 2.1–6.9 in cell lines and was not correlated with crizotinib sensitivity (IC50s 0.34–2.8 uM) (r=0.279, p=0.1764). Neither native, nor rearranged ALK copy number, nor percentage cells positive correlated with extra-central nervous system progression free survivalin ALK+ patients on crizotinib. Conclusions 8.5% of cases are below the established positivity threshold by ?5%. Further investigation of ALK by other diagnostic techniques in such cases may be warranted. Native ALK copy number increases alone are not associated with sensitivity to ALK inhibition in vitro. However, rare complex patterns of increased native ALK in patients should be studied further as atypical rearrangements contained within these may otherwise be missed. PMID:24022839

Camidge, D. Ross; Skokan, Margaret; Kiatsimkul, Porntip; Helfrich, Barbara; Lu, Xian; Baron, Anna E.; Schulte, Nathan; Maxson, DeLee; Aisner, Dara L.; Franklin, Wilbur A.; Doebele, Robert C.; Varella-Garcia, Marileila

2013-01-01

372

Effects of Corynebacterium parvum and BCG therapy on immune parameters in patients with disseminated melanoma a sequential study over 28 days. I. Changes in blood counts, serum immunoglobulins and lymphoid cell populations.  

PubMed Central

The effects of a single immunization of melanoma patients with BCG or C. parvum on the blood counts, serum immunoglobulin levels and lymphoid subpopulations were followed by multiple assays over 28 days. C. parvum produced a decrease in the white cell count, lymphocyte count and lymphoid T and sIg+ cell numbers, which recovered within 1 week; BCG did not produce such a marked depression. Both agents were associated with increases in T cell numbers and lymphocyte PHA blastogenesis after the first week; these declined to pre-immunization values by 3-4 weeks. The sIg-bearing cell subpopulation also increased after BCG. Different methods of expression the results were compared and the difficulties of immunological monitoring are discussed. PMID:428146

Thatcher, N; Swindell, R; Crowther, D

1979-01-01

373

Human amniotic fluid stem cells have a potential to recover ovarian function in mice with chemotherapy-induced sterility  

PubMed Central

Background Human amniotic fluid cells (hAFCs) may differentiate into multiple cell lineages and thus have a great potential to become a donor cell source for regenerative medicine. The ability of hAFCs to differentiate into germ cell and oocyte-like cells has been previously documented. Herein we report the potential use of hAFCs to help restore follicles in clinical condition involving premature ovarian failure. Results Human amniotic fluid was obtained via amniocentesis, yielding a subpopulation of cloned hAFCs that was able to form embryoid bodies (EBs) and differentiate into three embryonic germ layers. Moreover, culture of EBs in medium containing human follicular fluid (HFF) or a germ cell maturation factor cocktail (FAC), expressed germ cells markers such as BLIMP1, STELLA, DAZL, VASA, STRA8, SCP3, SCP1, and GDF9. Furthermore, one cell line was grown from clone cells transfected with lentivirus-GFP and displaying morphological characteristics of mesenchymal cells, had the ability to restore ovarian morphology following cell injection into the ovaries of mice sterilized by intraperitoneal injection of cyclophosphamide and busulphan. Restored ovaries displayed many follicle-enclosed oocytes at all stages of development, but no oocytes or follicles were observed in sterilized mice whose ovaries had been injected with medium only (control). Notably, identification of GFP-labeled cells and immunostaining with anti–human antigen-specific antibodies demonstrated that grafted hAFCs survived and differentiated into granulosa cells which directed oocyte maturation. Furthermore, labeling of ovarian tissue for anti-Müllerian hormone expression, a functional marker of folliculogenesis, was strong in hAFCs-transplanted ovaries but inexistent in negative controls. Conclusion These findings highlight the possibility of using human amniotic fluid-derived stem cells in regenerative medicine, in particular in the area of reproductive health. PMID:24006896

2013-01-01

374

Evaluation of clustering of new intramammary infections in the bovine udder, including the impact of previous infections, herd prevalence, and somatic cell count on their development.  

PubMed

Evidence in the literature exists to support the theory that mastitis and intramammary infection (IMI) tend to cluster within herds, within cows, and within quarters, facts which may have overarching ramifications on mastitis management in modern dairy herds. Most previous studies, however, have been carried out on prevalent IMI instead of new IMI (NIMI), although reducing incidence of NIMI is a major step toward controlling mastitis. The Canadian Bovine Mastitis Research Network (Saint-Hyacinthe, QC, Canada) has a large mastitis database derived from a 2-yr data collection on a national cohort of dairy farms, and data from this initiative were used to investigate the effect of clustering on the acquisition of NIMI. Longitudinal milk samplings of clinically normal udders taken over several 6-wk periods as well as samples from cows pre-dry-off and postcalving were used (n=73,772 quarter milk samples). Multilevel logistic models were used to evaluate the effect of location of IMI in quarters of the bovine udder previous to occurrence of an NIMI with Staphylococcus aureus, coagulase-negative staphylococci, and Corynebacterium spp. Several factors were investigated, including the number and location of quarters infected with the pathogen of interest before occurrence of an NIMI, the number of quarters infected with any other pathogen before occurrence of an NIMI (a measure of susceptibility), somatic cell count of the quarter before occurrence of an NIMI, somatic cell count of the other 3 quarters before occurrence of an NIMI, prevalence of the specific pathogen in the herd, and the average somatic cell count of the herd. The amount of variation occurring at different levels (herd, cow, and quarter) for the various pathogens was also calculated. The presence of an IMI in the ipsilateral quarter was associated with an elevated risk of an NIMI occurring for all pathogens investigated. Risk of an NIMI increased considerably as herd prevalence of the pathogen rose. Substantial clustering was found at all levels, with roughly equal amounts of variation found in all 3 levels for coagulase-negative staphylococci, most variation at the cow-level for Corynebacterium spp., and most variation found at the quarter-level for Staph. aureus. Simulation was used to calculate exact values of intraclass correlation coefficients to estimate clustering within cows and within quarters--these exact values were, for the most part, lower than estimates calculated using the latent variable approach, but also increased as pathogen prevalence and number of infections in a cow at the previous sampling increased. These results of these analyses can be used to inform approaches to preventing NIMI in modern dairy operations. PMID:23164233

Reyher, K K; Dohoo, I R; Muckle, C A

2013-01-01

375

Low thymic output in the 22q11.2 deletion syndrome measured by CCR9+CD45RA+ T cell counts and T cell receptor rearrangement excision circles  

PubMed Central

Thymic hypoplasia is a frequent feature of the 22q11.2 deletion syndrome, but we know little about patients' age-related thymic output and long-term consequences for their immune system. We measured the expression of T cell receptor rearrangement excision circles (TREC) and used flow cytometry for direct subtyping of recent thymic emigrant (RTE)-related T cells in 43 patients (aged 1–54 years; median 9 years) from all over Norway and in age-matched healthy controls. Thymic volumes were estimated by ultrasound in patients. TREC levels correlated well with RTE-related T cells defined by co-expression of CD3, CD45RA and CCR9 (r = 0·84) as well as with the CD4+ and CD8+ T cell subtypes. RTE-related T cell counts also paralleled age-related TREC reductions. CD45RA+ T cells correlated well with absolute counts of CD4+ (r = 0·87) and CD8+ (r = 0·75) RTE-related T cells. Apart from CD45RA? T cells, all T cell subsets were lower in patients than in controls. Thymic volumes correlated better with RTE-related cells (r = 0·46) than with TREC levels (r = 0·38). RTE-related T cells and TREC levels also correlated well (r = 0·88) in patients without an identifiable thymus. Production of RTEs is impaired in patients with a 22q11.2 deletion, and CCR9 appears to be a good marker for RTE-related T cells. PMID:20491792

Lima, K; Abrahamsen, T G; Foelling, I; Natvig, S; Ryder, L P; Olaussen, R W

2010-01-01

376

Kinetics of T-cell-based assays on cerebrospinal fluid and peripheral blood mononuclear cells in patients with tuberculous meningitis  

PubMed Central

Background/Aims The goal of this study was to monitor tuberculosis (TB)-specific T-cell responses in cerebrospinal fluid-mononuclear cells (CSF-MCs) and peripheral blood mononuclear cells (PBMCs) in patients with tuberculous meningitis (TBM) over the course of anti-TB therapy. Methods Adult patients (? 16 years) with TBM admitted to Asan Medical Center, Seoul, South Korea, were prospectively enrolled between April 2008 and April 2011. Serial blood or CSF samples were collected over the course of the anti-TB therapy, and analyzed using an enzyme-linked immunosorbent spot (ELISPOT) assay. Results Serial ELISPOT assays were performed on PBMCs from 17 patients (seven definite, four probable, and six possible TBM) and CSF-MC from nine patients (all definite TBM). The median number of interferon-gamma (IFN-?)-producing T-cells steadily increased during the first 6 months after commencement of anti-TB therapy in PBMCs. Serial CSF-MC ELISPOT assays revealed significant variability in immune responses during the first 6 weeks of anti-TB therapy, though early increases in CSF-MC ELISPOT results were associated with treatment failure or paradoxical response. Conclusions Serial analysis of PBMCs by ELISPOT during the course of treatment was ineffective for predicting clinical response. However, increases in TB-specific IFN-?-producing T-cells in CSF-MC during the early phase of anti-TB therapy may be predictive of clinical failure. PMID:25378978

Park, Ki-Ho; Lee, Mi Suk; Lee, Sang-Oh; Choi, Sang-Ho; Kim, Yang Soo; Woo, Jun Hee; Kang, Joong Koo; Lee, Sang-Ahm

2014-01-01

377

Monocyte count at diagnosis is a prognostic parameter in diffuse large B-cell lymphoma: results from a large multicenter study involving 1191 patients in the pre- and post-rituximab era  

PubMed Central

In this study we assessed the prognostic significance of absolute monocyte count and selected the best cut-off value at diagnosis in a large cohort of patients with diffuse large B-cell lymphoma. Data were retrieved for therapy-naïve patients with diffuse large B-cell lymphoma followed in Israel and Italy during 1993–2010. A final cohort of 1017 patients was analyzed with a median follow up of 48 months and a 5-year overall survival rate of 68%. The best absolute monocyte count cut-off level was 630/mm3 and the 5-year overall survival for patients with counts below this cut-off was 71%, wherea