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Sample records for erythrocyte count

  1. Relationship between electrophoretic mobility of erythrocytes and blood erythrocyte count in rats.

    PubMed

    Matyushichev, V B; Shamratova, V G

    2005-03-01

    A significant curvilinear relationship was found between erythrocyte count in rat blood and electrokinetic characteristics of these cells. Electrophoretic mobility of erythrocytes remained unchanged, slightly increased, or decreased with increasing cell count in the vascular bed depending on animal state. Excessive increase in the number of erythrocytes was accompanied by accumulation of cells with low electrophoretic mobility in the electric field.

  2. A rapid method for counting nucleated erythrocytes on stained blood smears by digital image analysis

    USGS Publications Warehouse

    Gering, E.; Atkinson, C.T.

    2004-01-01

    Measures of parasitemia by intraerythrocytic hematozoan parasites are normally expressed as the number of infected erythrocytes per n erythrocytes and are notoriously tedious and time consuming to measure. We describe a protocol for generating rapid counts of nucleated erythrocytes from digital micrographs of thin blood smears that can be used to estimate intensity of hematozoan infections in nonmammalian vertebrate hosts. This method takes advantage of the bold contrast and relatively uniform size and morphology of erythrocyte nuclei on Giemsa-stained blood smears and uses ImageJ, a java-based image analysis program developed at the U.S. National Institutes of Health and available on the internet, to recognize and count these nuclei. This technique makes feasible rapid and accurate counts of total erythrocytes in large numbers of microscope fields, which can be used in the calculation of peripheral parasitemias in low-intensity infections.

  3. Relationship between erythrocyte count and volume in humans and rats.

    PubMed

    Matyushichev, V B; Shamratova, V G; Savrasova, I V

    2000-09-01

    The mean corpuscular volume and concentration of blood erythrocytes in intact male rats are inversely related in the entire fluctuations range. In healthy men and women the correlation between these parameters is described by a parabola with alternating zones of positive and negative relationships. These covariations are unstable; in disease they change and sometimes are transformed into monotonous reciprocal correlations.

  4. Relationship between decrease of erythrocyte count and progression of periodontal disease in a rural Japanese population.

    PubMed

    Yamamoto, Tatsuo; Tsuneishi, Midori; Furuta, Michiko; Ekuni, Daisuke; Morita, Manabu; Hirata, Yukio

    2011-01-01

    Limited research has been reported on the relationship between periodontal status and erythrocyte parameters. In the present study, longitudinal data from health checkups are used to clarify the relationship between periodontal disease progression and changes in parameters of erythrocytes. In this prospective study, a total of 120 subjects (35 men and 85 women; age range: 30 to 63 years) participated in a comprehensive health screening and dental checkup in 2006 and 2007. Medical examinations, including anthropometric and manometric measurements, blood-chemistry tests, and oral examinations, were performed. Subjects with periodontal disease progression (i.e., the progression group) were defined based on the presence of ≥1 tooth demonstrating a longitudinal loss of periodontal attachment ≥3 mm or tooth loss during the study period. The Wilcoxon signed-rank, χ(2), Fisher exact, and Mann-Whitney U tests and stepwise logistic regression analysis were used for statistical analyses. The progression group comprised 30 subjects. A significant difference between the progression and non-progression groups was observed in changes of erythrocyte counts but not those of the body mass index, systolic and diastolic blood pressure, creatinine, fasting blood glucose, hemoglobin, hematocrit, triglycerides, high- and low-density lipoprotein cholesterol, total cholesterol, aspartate and alanine aminotransferase, and γ-glutamyl transpeptidase. Stepwise logistic regression analysis showed that the progression of periodontal disease was associated with a change (year 2007 minus year 2006) of erythrocyte counts (adjusted odds ratio = 0.970; P = 0.009) after adjusting for age at baseline. Within the limitations of this study, the progression of periodontal disease is associated with a decrease in erythrocyte counts in a rural Japanese population.

  5. Influence of counting methodology on erythrocyte ratios in the mouse micronucleus test.

    PubMed

    LeBaron, Matthew J; Schisler, Melissa R; Torous, Dorothea K; Dertinger, Stephen D; Gollapudi, B Bhaskar

    2013-04-01

    The mammalian erythrocyte micronucleus test is widely used to investigate the potential interaction of a test substance with chromosomes or mitotic apparatus of replicating erythroblasts. In addition to the primary endpoint, micronucleated erythrocyte frequency, the proportion of immature erythrocytes is measured to assess the influence of treatment on erythropoiesis. The guideline recommendation for an acceptable limit of the immature erythrocyte fraction of not < 20% of the controls was based on traditional scoring methods that consider RNA content. Flow-based sample analysis (e.g., MicroFlow®) characterizes a subpopulation of RNA-containing reticulocytes (RETs) based on CD71 (transferrin receptor) expression. As CD71+ cells represent a younger cohort of RETs, we hypothesized that this subpopulation may be more responsive than the RNA+ fraction for acute exposures. This study evaluated RET population in the peripheral blood of two strains of mice treated by oral gavage with three clastogens (cyclophosphamide, N-ethyl-N-nitrosourea, and methyl methanesulfonate). Although CD71+ frequencies correlated with RNA-based counts, the relative treatment-related reductions were substantially greater. Accordingly, when using the flow cytometry-based CD71+ values for scoring RETs in an acute treatment design, it is suggested that a target value ≥ 5% CD71+ reticulocytes (i.e., 95% depression in reticulocytes proportion) be considered as acceptable for a valid assay.

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

    SciTech Connect

    Konstom, M.A.; Tu'meh, S.; Wynne, J.; Beck, J.R.; Kozlowski, J.; Holman, B.L.

    1982-09-01

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

  7. [Automated measurement of reticulocyte count by flow cytometry. II: Analysis of the blood containing abnormal erythrocytes or giant platelets].

    PubMed

    Oyamatsu, T; Shimizu, N; Takeuchi, K; Yamamoto, M; Kawai, Y; Watanabe, K; Iri, H

    1989-07-01

    We have examined the influence of erythrocytes containing inclusion bodies, nucleated red cells or giant platelets on the measurement of reticulocyte count by automated machine, R-1000. Correlation of the reticulocyte count between automated and conventional method was extremely good in the blood containing red cells with Jolly bodies, Pappenheimer bodies or basophilic stippling . However, correlation was poor when the sample contained the nucleated red cells. Reticulocyte count was decreased in the blood with significant amounts of nucleated red cells. Since nucleated red cells themselves are not counted as reticulocytes in the machine, this was considered to be due to increased young reticulocytes which frequently appeared with nucleated red cells. Both cold agglutinated red cells and giant platelets apparently influenced the reticulocyte count by the R-1000. These results suggest that red cells with Jolly bodies, Pappenheimer bodies or basophilic stippling do not influence the automatic counting of reticulocytes. Although nucleated red cells, cold agglutinated red cells and giant platelets affected the reticulocyte count, the machine shows abnormal flags in most of above cases (except highly agglutinated red cells), so that one can recount reticulocytes by conventional method. We conclude the machine can safely count the reticulocytes even in the blood containing abnormal red cells or platelets.

  8. Effect of exercise on erythrocyte count and blood activity concentration after /sup 99m/Tc in vivo red blood cell labeling

    SciTech Connect

    Konstam, M.A.; Tu'meh, S.; Wynne, J.; Beck, J.R.; Kozlowski, J.; Holman, B.L.

    1982-09-01

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

  9. [Plasma and erythrocyte membrane phospholipids in children with Plasmodium falciparum malaria: relation to blood parasite counts and lactate levels].

    PubMed

    Abessolo, Félix Ovono; Nguélé, Jean Calvin; Legault, Estelle; Ngou-Milama, Edouard

    2009-01-01

    Plasmodium falciparum infection modifies the distribution of phospholipids on both sides of the erythrocyte-plasma membrane. We sought to understand the action of the parasite on both plasma and membrane phospholipids and their relation to parasite counts in the blood. We conducted this study from 1 February through 30 June 2007 in the Malaria Clinical Research Unit of the Libreville General Hospital Centre, measuring phospholipids in plasma and erythrocyte membranes of children affected by P. falciparum malaria and assessing their relation to blood parasites, lactates and hemoglobin. The study included 60 children. The mean blood parasite count was 1.07 x 105 per ml. Mean plasma phospholipids were 2.6+/-0.9 mmol/l and lactates 4.7+/-3.7 mmol/l. At the membrane level, we found 2.8+/-1.2 mmol/l of phospholipids per 106 red blood cells. Phospholipids in these two compartments were positively correlated with one another (r=0.392; p=0.023). Plasma phospholipids were also correlated positively with hemoglobin (r=0.36; p=0.032) and negatively with the number of parasites (r=0.442; p=0.002). The latter, in turn, was positively correlated with lactates (r=0.527; p=0.022) and negatively with membrane phospholipids (r=0.542; p=0.006). The mean hemoglobin level (8.9 g/dl) suggests permanent blood loss in these children, although they may be considered new malaria patients. The relations revealed here between membrane phospholipids, blood parasite counts, and lactate levels are due to the increased energy needs of erythrocytes, but also to the need to synthesize substances intended for cellular defense. P. falciparum thus leads to a decrease in plasma phospholipids, which could be estimated and correlated with blood parasites in a study with a wider range of parasite counts.

  10. The utility of erythrocyte sedimentation rate values and white blood cell counts after spinal deformity surgery in the early (≤3 months) post-operative period.

    PubMed

    Kuhn, Margaret G; Lenke, Lawrence G; Bridwell, Keith H; O'Donnell, June C; Luhmann, Scott J

    2012-03-01

    The erythrocyte sedimentation rate (ESR) and white blood cell (WBC) count are frequently obtained in the work-up of post-operative fever. However, their diagnostic utility depends upon comparison with normative peri-operative trends which have not yet been described. The purpose of this study is to define a range of erythrocyte sedimentation rates and white blood cell counts following spinal instrumentation and fusion in non-infected patients. Seventy-five patients underwent spinal instrumentation and fusion. The erythrocyte sedimentation rate and white blood cell count were recorded pre-operatively, at 3 and 7 days post-operatively, and at 1 and 3 months post-operatively. Both erythrocyte sedimentation rate and white blood cell count trends demonstrated an early peak, followed by a gradual return to normal. Peak erythrocyte sedimentation rates occurred within the first week post-operatively in 98% of patients. Peak white blood cell counts occurred with the first week in 85% of patients. In the absence of infection, the erythrocyte sedimentation rate was abnormally elevated in 78% of patients at 1 month and in 53% of patients at 3 months post-operatively. The white blood cell count was abnormally elevated in only 6% of patients at 1 month post-operatively. Longer surgical time was associated with elevated white cell count at 1 week post-operatively. The fusion of more vertebral levels had a negative relationship with elevated erythrocyte sedimentation rate at 1 week post-operatively. The anterior surgical approach was associated with significantly lower erythrocyte sedimentation rate at 1 month post-operatively and with lower white cell count at 1 week post-operatively. In non-infected spinal fusion surgeries, erythrocyte sedimentation rates are in the abnormal range in 78% of patients at 1 month and in 53% of patients at 3 months post-operatively, suggesting that the erythrocyte sedimentation rate is of limited diagnostic value in the early post

  11. Estimating leukocyte, platelet, and erythrocyte counts in rats by blood smear examination.

    PubMed

    Durbin, Cathy; Guo, Kevin; Hoffman, Wherly; Eric Schultze, A; White, Sandy

    2009-06-01

    The CBC is an essential test for assessing the health of rats used in drug development studies. Because of limited blood volume, estimates of cell counts from a blood smear would be valuable when other analytical methods of enumerating cells are not possible or available. The purpose of this study was to develop a statistical model to accurately estimate WBC, platelet (PLT), and RBC counts in blood smears from rats. Blood smears and quantitative cell counts were obtained from vehicle-treated male and female Fischer 344 rats (n=65) involved in a variety of studies. The numbers of WBCs, PLTs, and RBCs were estimated in 10 fields in the monolayer of smears using x 20 (WBC) or x 100 (PLT, RBC) objectives. Using a statistical model and the quantitative cell counts obtained on an ADVIA 120 hematology analyzer, formulas were developed to predict the quantitative counts from the estimates. Data were log-transformed before analysis. A formula was derived using the slope and intercept of the regression line between cell estimates and ADVIA counts to predict WBC, PLT, and RBC counts based only on estimates. A second formula was developed for situations in which limited quantitative analyses may be available, and resulted in even more accurately predicted counts from smear estimates. The formulas developed in this study can be a valuable tool in estimating cell counts from a blood smear when cell counting instruments are not available or when an instrument cell count needs to be verified. These formulas may be useful in the assessment of rat blood in discovery and lead optimization studies.

  12. Dynamics of erythrocyte count, hemoglobin, and catalase activity in rat blood in hypokinesia, muscular activity and restoration

    NASA Technical Reports Server (NTRS)

    Taneyeva, G. V.; Potapovich, G. M.; Voloshko, N. A.; Uteshev, A. B.

    1980-01-01

    Tests were conducted to prove that muscular exertion (in this instance swimming) of different duration and intensity, as well as hypodynamia, result in an increase of hemoglobin and number of red blood cells in peripheral blood rats. Catalase activity increased with an increase in the duration of swimming, but only up to 6 hr; with 7-9 hr of swimming as well as in hypodynamia, catalase activity decreased. It was also observed that under hypodynamia as well as in 3, 5 and 6 hr exertion (swimming) the color index of blood decreased. Pressure chamber treatment (for 8 min each day for one week), alternating a 2 min negative pressure up to 35 mm Hg with 1 min positive pressure, increased the erythrocyte count and hemoglobin content.

  13. Flow Cytometry Analysis Using Sysmex UF-1000i Classifies Uropathogens Based on Bacterial, Leukocyte, and Erythrocyte Counts in Urine Specimens among Patients with Urinary Tract Infections

    PubMed Central

    Rydén, Patrik

    2014-01-01

    Urinary tract infections (UTIs) are the second most common bacterial infection. Urine culture is the gold standard for diagnosis, but new techniques, such as flow cytometry analysis (FCA), have been introduced. The aim of the present study was to evaluate FCA characteristics regarding bacteriuria, leukocyturia, and erythrocyturia in relation to cultured uropathogens in specimens from patients with a suspected UTI. We also wanted to evaluate whether the FCA characteristics can identify uropathogens prior to culture. From a prospective study, 1,587 consecutive urine specimens underwent FCA prior to culture during January and February 2012. Outpatients and inpatients (79.6% and 19.4%, respectively) were included, of whom women represented 67.5%. In total, 620 specimens yielded growth, of which Escherichia coli represented 65%, Enterococcus spp. 8%, Klebsiella spp. 7%, and Staphylococcus spp. 5%. For the uropathogens, the outcome of FCA was compared against the results for specimens with E. coli and those with a negative culture. E. coli had high bacterial (median, 17,914/μl), leukocyte (median, 348/μl), and erythrocyte (median, 23/μl) counts. With the exception of Klebsiella spp., the majority of the uropathogens had considerable or significantly lower bacterial counts than that of E. coli. High leukocyte counts were found in specimens with Staphylococcus aureus, Proteus mirabilis, Pseudomonas aeruginosa, and group C streptococci. Elevated erythrocyte counts were found for P. vulgaris, P. aeruginosa, and group C streptococci, as well as for Staphylococcus saprophyticus. In essence, FCA adds new information about the bacterial, leukocyte, and erythrocyte counts in urine specimens for different uropathogens. Based on FCA characteristics, uropathogens can be classified and identified prior to culture. E. coli and Klebsiella spp. have similar FCA characteristics. PMID:25472486

  14. Flow cytometry analysis using sysmex UF-1000i classifies uropathogens based on bacterial, leukocyte, and erythrocyte counts in urine specimens among patients with urinary tract infections.

    PubMed

    Monsen, Tor; Rydén, Patrik

    2015-02-01

    Urinary tract infections (UTIs) are the second most common bacterial infection. Urine culture is the gold standard for diagnosis, but new techniques, such as flow cytometry analysis (FCA), have been introduced. The aim of the present study was to evaluate FCA characteristics regarding bacteriuria, leukocyturia, and erythrocyturia in relation to cultured uropathogens in specimens from patients with a suspected UTI. We also wanted to evaluate whether the FCA characteristics can identify uropathogens prior to culture. From a prospective study, 1,587 consecutive urine specimens underwent FCA prior to culture during January and February 2012. Outpatients and inpatients (79.6% and 19.4%, respectively) were included, of whom women represented 67.5%. In total, 620 specimens yielded growth, of which Escherichia coli represented 65%, Enterococcus spp. 8%, Klebsiella spp. 7%, and Staphylococcus spp. 5%. For the uropathogens, the outcome of FCA was compared against the results for specimens with E. coli and those with a negative culture. E. coli had high bacterial (median, 17,914/μl), leukocyte (median, 348/μl), and erythrocyte (median, 23/μl) counts. With the exception of Klebsiella spp., the majority of the uropathogens had considerable or significantly lower bacterial counts than that of E. coli. High leukocyte counts were found in specimens with Staphylococcus aureus, Proteus mirabilis, Pseudomonas aeruginosa, and group C streptococci. Elevated erythrocyte counts were found for P. vulgaris, P. aeruginosa, and group C streptococci, as well as for Staphylococcus saprophyticus. In essence, FCA adds new information about the bacterial, leukocyte, and erythrocyte counts in urine specimens for different uropathogens. Based on FCA characteristics, uropathogens can be classified and identified prior to culture. E. coli and Klebsiella spp. have similar FCA characteristics.

  15. [Relationship between white blood cell count, neutrophils ratio and erythrocyte sedimentation rate and short clinical outcomes among patients with acute ischemic stroke at hospital admission].

    PubMed

    Ye, Jia-kai; Zhang, Jin-tao; Kong, Yan; Xu, Tan; Zou, Ting-ting; Zhang, Yong-hong; Zhang, Shao-yan

    2012-09-01

    To investigate the relationship between white blood cell count, neutrophils ratio and erythrocyte sedimentation rate and short outcomes among patients with acute ischemic stroke at admission to the hospital. A total of 2675 acute ischemic stroke patients were included in this study. Data on demographic characteristics, life style, history of disease, white blood cell count (WBC), neutrophils ratio (NEUR), erythrocyte sedimentation rate (ESR) and clinical outcomes were collected for all the participants. Poor clinical outcome was defined as neurologic deficiency (NIHSS ≥ 5) at discharge or death during hospitalization. White blood cell count, neutrophils ratio and erythrocyte sedimentation rate were higher in patients with poor outcome than in those without clinical outcome. According to the quartile range, WBC, NEUR and ESR were divided into four levels at admission. After adjustment for multivariate, compared with WBC ≤ 5.6× 10(9)/L, the odds ratio (95% confidence intervals) of poor outcome with ≥ 8.7×10(9)/L was 1.883 (1.306 - 2.716). When compared with NEUR ≤ 0.56, the odds ratio (95% confidence intervals) of poor outcome with 0.57 - 0.64 and with ≥ 0.74 were 1.572 (1.002 - 2.466) and 2.577 (1.698 - 3.910), respectively. When compared with ESR ≤ 4 mm/h, the odds ratio (95% confidence intervals) of poor outcome with ≥ 17 mm/h was 2.426 (1.233 - 4.776). Elevated WBC count and NEUR at admission were significantly and positively associated with poor clinical outcomes among patients with acute ischemic stroke (trend test P < 0.05). Elevated ESR was not significantly or positively associated with poor clinical outcomes among patients with acute ischemic stroke (trend test P > 0.05). There appeared associations between WBC, NEUR, ESR and poor outcome among patients with acute ischemic stroke at admission to the hospital. Both elevated WBC count and NEUR showed significantly positive association with poor clinical outcomes among patients with acute

  16. Erythrocytes and the regulation of human skeletal muscle blood flow and oxygen delivery: role of erythrocyte count and oxygenation state of haemoglobin

    PubMed Central

    González-Alonso, José; Mortensen, Stefan P; Dawson, Ellen A; Secher, Niels H; Damsgaard, Rasmus

    2006-01-01

    Blood flow to dynamically contracting myocytes is regulated to match O2 delivery to metabolic demand. The red blood cell (RBC) itself functions as an O2 sensor, contributing to the control of O2 delivery by releasing the vasodilators ATP and S-nitrosohaemoglobin with the offloading of O2 from the haemoglobin molecule. Whether RBC number is sensed remains unknown. To investigate the role of RBC number, in isolation and in combination with alterations in blood oxygenation, on muscle and systemic perfusion, we measured local and central haemodynamics during one-legged knee-extensor exercise (∼50% peak power) in 10 healthy males under conditions of normocythaemia (control), anaemia, anaemia + plasma volume expansion (PVX), anaemia + PVX + hypoxia, polycythaemia, polycythaemia + hyperoxia and polycythaemia + hypoxia, which changed either RBC count alone or both RBC count and oxyhaemoglobin. Leg blood flow (LBF), cardiac output (Q) and vascular conductance did not change with either anaemia or polycythaemia alone. However, LBF increased with anaemia + PVX (28 ± 4%) and anaemia + PVX + hypoxia (46 ± 6%) and decreased with polycythaemia + hyperoxia (18 ± 5%). LBF and Q with anaemia + PVX + hypoxia (8.0 ± 0.5 and 15.8 ± 0.7 l min−1, respectively) equalled those during maximal knee-extensor exercise. Collectively, LBF and vascular conductance were intimately related to leg arterial–venous (a–v) O2 difference (r2 = 0.89–0.93; P < 0.001), suggesting a pivotal role of blood O2 gradients in muscle microcirculatory control. The systemic circulation accommodated to the changes in muscle perfusion. Our results indicate that, when coping with severe haematological challenges, local regulation of skeletal muscle blood flow and O2 delivery primarily senses alterations in the oxygenation state of haemoglobin and, to a lesser extent, alterations in the number of RBCs and haemoglobin molecules. PMID:16439430

  17. Erythrocytes and the regulation of human skeletal muscle blood flow and oxygen delivery: role of erythrocyte count and oxygenation state of haemoglobin.

    PubMed

    González-Alonso, José; Mortensen, Stefan P; Dawson, Ellen A; Secher, Niels H; Damsgaard, Rasmus

    2006-04-01

    Blood flow to dynamically contracting myocytes is regulated to match O(2) delivery to metabolic demand. The red blood cell (RBC) itself functions as an O(2) sensor, contributing to the control of O(2) delivery by releasing the vasodilators ATP and S-nitrosohaemoglobin with the offloading of O(2) from the haemoglobin molecule. Whether RBC number is sensed remains unknown. To investigate the role of RBC number, in isolation and in combination with alterations in blood oxygenation, on muscle and systemic perfusion, we measured local and central haemodynamics during one-legged knee-extensor exercise ( approximately 50% peak power) in 10 healthy males under conditions of normocythaemia (control), anaemia, anaemia + plasma volume expansion (PVX), anaemia + PVX + hypoxia, polycythaemia, polycythaemia + hyperoxia and polycythaemia + hypoxia, which changed either RBC count alone or both RBC count and oxyhaemoglobin. Leg blood flow (LBF), cardiac output (Q) and vascular conductance did not change with either anaemia or polycythaemia alone. However, LBF increased with anaemia + PVX (28 +/- 4%) and anaemia + PVX + hypoxia (46 +/- 6%) and decreased with polycythaemia + hyperoxia (18 +/- 5%). LBF and Q with anaemia + PVX + hypoxia (8.0 +/- 0.5 and 15.8 +/- 0.7 l min(-1), respectively) equalled those during maximal knee-extensor exercise. Collectively, LBF and vascular conductance were intimately related to leg arterial-venous (a-v) O(2) difference (r(2)= 0.89-0.93; P < 0.001), suggesting a pivotal role of blood O(2) gradients in muscle microcirculatory control. The systemic circulation accommodated to the changes in muscle perfusion. Our results indicate that, when coping with severe haematological challenges, local regulation of skeletal muscle blood flow and O(2) delivery primarily senses alterations in the oxygenation state of haemoglobin and, to a lesser extent, alterations in the number of RBCs and haemoglobin molecules.

  18. Development and evaluation of a novel in-clinic automated hematology analyzer, ProCyte Dx, for canine erythrocyte indices, leukogram, platelet counts and reticulocyte counts.

    PubMed

    Fujino, Yasuhito; Nakamura, Yoichi; Matsumoto, Hideaki; Fukushima, Kenjiro; Takahashi, Masashi; Ohno, Koichi; Tsujimoto, Hajime

    2013-11-01

    A novel hematology analyzer for small animal medicine, ProCyte Dx, was developed from combination of the fluorescence laser flow cytometry and laminar flow impedance technologies, and its accuracy was evaluated by comparing with the conventional impedance-based hematology analyzer, pocH-100iV Diff, or microscopic manual cell counting methods with staining blood smears in the canine blood. Blood samples of 59 dogs were hematologically analyzed and compared by Pearson's correlation coefficients. Analyses between the two analyzers showed excellent correlation in RBC (r=0.998), HGB (r=0.999), HCT (r=0.998), MCV (r=0.994), MCH (r=0.974), MCHC (r=0.906), WBC (r=0.998) and PLT (r=0.993). Analyses between ProCyte Dx and microscopic manual counting results showed excellent correlation in neutrophils (r=0.920), lymphocytes (r=0.913) and reticulocyte percentages (r=0.924), good correlation in eosinophils (r=0.815) and reticulocyte numbers (r=0.850) and fair correlation in monocytes (r=0.770). The present study indicates that ProCyte Dx is acceptably accurate and can be a powerful tool for canine clinical medicine.

  19. Development and Evaluation of a Novel In-Clinic Automated Hematology Analyzer, ProCyte Dx, for Canine Erythrocyte Indices, Leukogram, Platelet Counts and Reticulocyte Counts

    PubMed Central

    FUJINO, Yasuhito; NAKAMURA, Yoichi; MATSUMOTO, Hideaki; FUKUSHIMA, Kenjiro; TAKAHASHI, Masashi; OHNO, Koichi; TSUJIMOTO, Hajime

    2013-01-01

    ABSTRACT A novel hematology analyzer for small animal medicine, ProCyte Dx, was developed from combination of the fluorescence laser flow cytometry and laminar flow impedance technologies, and its accuracy was evaluated by comparing with the conventional impedance-based hematology analyzer, pocH-100iV Diff, or microscopic manual cell counting methods with staining blood smears in the canine blood. Blood samples of 59 dogs were hematologically analyzed and compared by Pearson’s correlation coefficients. Analyses between the two analyzers showed excellent correlation in RBC (r=0.998), HGB (r=0.999), HCT (r=0.998), MCV (r=0.994), MCH (r=0.974), MCHC (r=0.906), WBC (r=0.998) and PLT (r=0.993). Analyses between ProCyte Dx and microscopic manual counting results showed excellent correlation in neutrophils (r=0.920), lymphocytes (r=0.913) and reticulocyte percentages (r=0.924), good correlation in eosinophils (r=0.815) and reticulocyte numbers (r=0.850) and fair correlation in monocytes (r=0.770). The present study indicates that ProCyte Dx is acceptably accurate and can be a powerful tool for canine clinical medicine. PMID:23811854

  20. Menstruation does not cause anemia: endometrial thickness correlates positively with erythrocyte count and hemoglobin concentration in premenopausal women.

    PubMed

    Clancy, Kathryn B H; Nenko, Ilona; Jasienska, Grazyna

    2006-01-01

    Menstruation has often been cited as a risk factor for iron-deficiency anemia. This study tested whether normal, premenopausal women's luteal endometrial thickness (ET) was associated with their red blood cell count (RBC) and hemoglobin concentrations (Hg), and therefore whether a high ET put women at risk for anemia. Endometrial thickness can be considered a reasonable proxy for menstrual blood loss in normal women. Twenty-six healthy women from the Mogielica Human Ecology Study Site in Poland, aged 20-40 years (29 +/- 5.3 years, mean +/- SD), were selected. Subjects' ET was measured by transvaginal ultrasound in the luteal phase of the menstrual cycle, and their red blood cell count and hemoglobin concentrations were measured by fasting morning blood samples. Controlling for day of ET measurement, RBC and Hg were positively correlated with ET (r(2) = 0.24, P = 0.05; r(2) = 0.25, P = 0.04, respectively). We propose that, contrary to popular understanding, a thicker endometrium suggests greater iron reserves, rather than greater risk for anemia, in healthy women.

  1. Interaction between leukocytes and erythrocytes in the human retina: effects of pentoxifylline on hyperoxia-induced vasoconstriction during increased neutrophil counts.

    PubMed

    Told, Reinhard; Fuchsjäger-Mayrl, Gabriele; Wolzt, Michael; Popa-Cherecheanu, Alina; Schmetterer, Leopold; Garhofer, Gerhard

    2014-03-01

    Pentoxifylline, a nonselective phosphodiesterase inhibitor, shows vasodilator effects in certain vascular beds and reduces blood viscosity. We have previously shown that under states of vasoconstriction an interaction between circulating erythrocytes and leukocytes may play a role in the control of blood flow. The reason for this observation is not entirely clear but may be related to a mechanical interaction between red and white blood cells. In the present study we hypothesized that pentoxifylline may alter this interaction during oxygen-induced vasoconstriction. 24 healthy male subjects participated in this double masked, randomized, placebo-controlled 2 way cross over trial. In order to increase white blood cell count (WBC) count, 300 μg of G-CSF was administered intravenously. Vasoconstriction of retinal vessels was induced by oxygen inhalation. 400mg of pentoxifylline or placebo was infused at two different study days. White blood cell flux was assessed with the blue-field entoptic technique. Vessel calibers were measured with a dynamic vessel analyzer (DVA) and red blood cell velocity (RBCV) was determined with laser Doppler velocimetry (LDV). Retinal blood flow was calculated based on retinal vessel diameters and RBCV. Administration of G-CSF induced a significant increase in WBC, both in the placebo and the pentoxifylline group (p<0.01 for both groups). Retinal vessel diameter, RBCV, calculated retinal blood flow and white blood cell flow were not altered by administration of pentoxifylline. Hyperoxia induced a pronounced decrease in retinal blood flow parameters. No difference was observed between groups during oxygen breathing in vessel diameters (p=0.54), RBCV (p=0.34), calculated retinal blood flow (p=0.3) and white blood cell flow (p=0.26). Our data indicate that short time administration of pentoxifylline does not alter the oxygen-induced effect on ocular blood flow parameters during leukocytosis. Whether long-term treatment could improve retinal

  2. ESR (Erythrocyte Sedimentation Rate) Test

    MedlinePlus

    ... AACC products and services. Advertising & Sponsorship: Policy | Opportunities Erythrocyte Sedimentation Rate (ESR) Share this page: Was this ... with conditions that inhibit the normal sedimentation of red blood cells, such as a high red blood cell count ( ...

  3. Erythrocyte rheology.

    PubMed

    Stuart, J

    1985-09-01

    Erythrocyte deformability was formerly measured by its contribution to whole blood viscosity. It is now more commonly measured by filtration of erythrocytes through, or aspiration into, pores of 3-5 microns diameter and by the measurement of shear induced erythrocyte elongation using laser diffractometry. Recent improvements in the technology for erythrocyte filtration have included the removal of acute phase reactants from test erythrocyte suspensions, ultrasonic cleaning and reuse of filter membranes, awareness of the importance of mean cell volume as a determinant of flow through 3 microns diameter pores, and the ability to detect subpopulations of less deformable erythrocytes. Measurements of erythrocyte elongation by laser diffractometry, using the Ektacytometer, are also influenced by cell size and need to be corrected for mean cell volume. These advances have greatly improved the sensitivity and specificity of rheological methods for measuring the deformability of erythrocytes and for investigating the mode of action of rheologically active drugs.

  4. Can Ratio of Neutrophil-to-Lymphocyte Count and Erythrocyte Sedimentation Rate in Diabetic Foot Infecti on Predict Osteomyelitis and/or Amputation?

    PubMed Central

    Yapıcı, Oktay; Berk, Hande; Öztoprak, Nefise; Seyman, Derya; Tahmaz, Alper; Merdin, Alparslan

    2017-01-01

    The aim of this study was to search for any relations between the neutrophil-to-lymphocyte ratio (NLR) and the development of osteomyelitis and the need for amputation in patients with diabetic foot infection (DFI). All data included DFI patients who were hospitalized in our Infectious Diseases Clinic between 2012 and 2015 and who were classified according to International Classification Disease Code System. 75 patients were analyzed in the study. The DFI patients were stratified into 3 groups of whom had amputation procedure, whom had only debridement/drainage procedure and whom had any surgery procedure. Sidac post hoc analysis was used to perform the effects of NLR, C-reactive protein, erythrocyte sedimentation rate and glycosylated hemoglobin on the surgery procedure status. The DFI patients were also stratified into two another separate group for another analysis to search for the effect of NLR values on the development of osteomyelitis. The mean value of NLR in the amputated patients’ group (15.7±10.3 was significantly higher than those with debridement procedure (9.9±5.6) and those without any surgery (6.0±2.8) (P=0.001). NLR values were also found significantly higher in patients with osteomyelitis in the second analysis (P=0.004). In this study, the NLR was found to have a predictive value on the development of osteomyelitis and on the progression to amputation in patients with DFI. PMID:28286632

  5. Conjugated Bilirubin Triggers Anemia by Inducing Erythrocyte Death

    PubMed Central

    Lang, Elisabeth; Gatidis, Sergios; Freise, Noemi F; Bock, Hans; Kubitz, Ralf; Lauermann, Christian; Orth, Hans Martin; Klindt, Caroline; Schuier, Maximilian; Keitel, Verena; Reich, Maria; Liu, Guilai; Schmidt, Sebastian; Xu, Haifeng C; Qadri, Syed M; Herebian, Diran; Pandyra, Aleksandra A; Mayatepek, Ertan; Gulbins, Erich; Lang, Florian; Häussinger, Dieter; Lang, Karl S; Föller, Michael; Lang, Philipp A

    2015-01-01

    Hepatic failure is commonly associated with anemia, which may result from gastrointestinal bleeding, vitamin deficiency, or liver-damaging diseases, such as infection and alcohol intoxication. At least in theory, anemia during hepatic failure may result from accelerated clearance of circulating erythrocytes. Here we show that bile duct ligation (BDL) in mice leads to severe anemia despite increased reticulocyte numbers. Bilirubin stimulated suicidal death of human erythrocytes. Mechanistically, bilirubin triggered rapid Ca2+ influx, sphingomyelinase activation, formation of ceramide, and subsequent translocation of phosphatidylserine to the erythrocyte surface. Consistent with our in vitro and in vivo findings, incubation of erythrocytes in serum from patients with liver disease induced suicidal death of erythrocytes in relation to their plasma bilirubin concentration. Consistently, patients with hyperbilirubinemia had significantly lower erythrocyte and significantly higher reticulocyte counts compared to patients with low bilirubin levels. Conclusion: Bilirubin triggers suicidal erythrocyte death, thus contributing to anemia during liver disease. (Hepatology 2015;61:275–284) PMID:25065608

  6. Effects of nickel chloride on the erythrocytes and erythrocyte immune adherence function in broilers.

    PubMed

    Li, Jian; Wu, Bangyuan; Cui, Hengmin; Peng, Xi; Fang, Jing; Zuo, Zhicai; Deng, Junliang; Wang, Xun; Tang, Kun; Yin, Shuang

    2014-11-01

    This study was conducted to investigate the immune adherence function of erythrocytes and erythrocyte induced by dietary nickel chloride (NiCl2) in broilers fed on a control diet and three experimental diets supplemented with 300, 600, and 900 mg/kg NiCl2 for 42 days. Blood samples were collected from five broilers in each group at 14, 28, and 42 days of age. Changes of erythrocyte parameters showed that total erythrocyte count (TEC), hemoglobin (Hb) contents, and packed cell volume (PCV) were significantly lower (p < 0.05 or p < 0.01) and erythrocyte osmotic fragility (EOF) was higher (p < 0.05 or p < 0.01) in the 600 and 900 mg/kg groups at 28 and 42 days of age than those in the control group, and the sodium-potassium adenosine triphosphatase (Na(+)/K(+)-ATPase) and calcium adenosine triphosphatase (Ca(2+)-ATPase) activities were significantly decreased (p < 0.05 or p < 0.01) in the NiCl2-treated groups. The results of erythrocyte immune adherence function indicated that erythrocyte C3b receptor rosette rate (E-C3bRR) was significantly decreased (p < 0.05 or p < 0.01) in the 600 and 900 mg/kg groups and in the 300 mg/kg group at 42 days of age, whereas the erythrocyte immune complex rosette rate (E-ICRR) was markedly increased (p < 0.05 or p < 0.01) in the 300, 600, and 900 mg/kg groups at 28 and 42 days of age. It was concluded that dietary NiCl2 in excess of 300 mg/kg caused anemia and impaired the erythrocytic integrity, erythrocytic ability to transport oxygen, and erythrocyte immune adherence function in broilers. Impairment of the erythrocytes and erythrocyte immune adherence function was one of main effect mechanisms of NiCl2 on the blood function.

  7. Antithrombin Activity of Erythrocyte Microvesicles.

    PubMed

    Levin, G Ya; Sukhareva, E G

    2017-04-01

    Coagulation and optical (based on chromogenic substrate) methods were employed to examine antithrombin activity of erythrocytes and erythrocyte-derived microvesicles isolated days 7, 14, 21, and 28 on erythrocyte storage. The erythrocyte-derived microvesicles decelerated fibrin clot formation from fibrinogen in the presence of exogenous thrombin both with and without heparin. Microvesicles reduced optical density of chromogenic substrate. These data suggest that erythrocyte-derived microvesicles display a prominent antithrombin activity, which significantly increases during erythrocyte storage.

  8. Response of the rat erythrocyte to ozone exposure

    NASA Technical Reports Server (NTRS)

    Larkin, E. C.; Kimzey, S. L.; Siler, K.

    1978-01-01

    Sprague-Dawley rats were exposed to high (6-8 ppm) and moderate (1.5 ppm) amounts of ozone (O3) for various time periods. Response of the rat erythrocyte to ozone was monitored with red blood cell potassium (rubidium) influx studies, with storage stress combined with ultrastructural studies and with levels of erythrocyte glutathione peroxidase and superoxide dismutase. Erythrocytes of rats exposed to O3 showed no significant changes either in their potassium influx or in their glutathione peroxidase and superoxide dismutase activities compared to controls. Erythrocyte differential counts on O3-exposed animals showed significant changes initially as well as following storage stress compared to controls. Rats exposed to 8 ppm O3 for 4 h showed a marked increase in echinocytes. These consistent transformations from discocytes to echinocytes following O3 exposure suggest latent erythrocyte damage has occurred.

  9. [Anemia and thrombocytopenia in malaria. II. Mean erythrocyte and thrombocyte count in 45 patients with malaria observed in the Clinic of the Institute of Maritime and Tropical Medicine in Gdynia].

    PubMed

    Górski, J; Nahorski, W; Goljan, J; Felczak-Korzybska, I; Myjak, P

    1994-01-01

    In the years 1987-92, 45 patients with malaria were hospitalized in our clinic. All the patients were before staying in tropical countries. 30 patients were infected with PI. falciparum while the rest - with other species of this parasite. From among numerous different symptoms of malaria, in the majority of the patients profound anaemia and thrombocytopenia occurred. The simultaneously observed drop in the erythrocyte and blood platelet levels confirms that the destruction pathways of them in patients with malaria may be similar, as are the reasons for ineffective erythro- and thrombocytopoiesis. Anaemia and thrombocytopenia gradually subsided in the course of the therapy, as the parasitemia decreased and the state of the patients improved.

  10. Effects of high dietary fluorine on erythrocytes and erythrocyte immune adherence function in broiler chickens.

    PubMed

    Deng, Yubing; Cui, Hengmin; Peng, Xi; Fang, Jing; Zuo, Zhicai; Deng, Junliang; Luo, Qin

    2013-11-01

    Fluoride can exert toxic effects on soft tissues, giving rise to a broad array of symptoms and pathological changes. The aim of this study was to investigate on erythrocytes and erythrocyte immune adherence function in broiler chickens fed with high fluorine (F) diets by measuring the total erythrocyte count (TEC), the contents of hemoglobin (Hb), packed cell volumn (PCV), erythrocyte osmotic fragility (EOF), erythrocyte C3b receptor rosette rate (E-C3bRR), and erythrocyte immune complex rosette rate (E-ICRR). A total of 280 1-day-old healthy avian broiler chickens were randomly allotted into four equal groups of 70 birds each and fed with a corn-soybean basal diet containing 22.6 mg F/kg (control group) or same basal diets supplemented with 400, 800, and 1,200 mg F/kg (high F groups I, II, and III) in the form of sodium fluoride for 42 days. Blood samples were collected for the abovementioned parameters analysis at 14, 28, and 42 days of age during the experiment. The experimental results indicated that TEC, Hb, and PCV were significantly lower (p < 0.05 or p < 0.01), and EOF was higher (p < 0.05 or p < 0.01) in the high F groups II and III than that in the control group from 14 to 42 days of age. The E-C3bRR was significantly decreased (p < 0.01) in the three high F groups, whereas the E-ICRR was markedly increased (p < 0.01) in the high F groups II and III from 14 to 42 days of age. It was concluded that dietary F in the range of 800 to 1, 200 mg/kg could significantly cause anemia and impair the integrity of erythrocyte membrane, the transport capacity of oxygen and carbon dioxide, and erythrocyte immune adherence function in broiler chickens.

  11. Storage of Erythrocytes Induces Suicidal Erythrocyte Death.

    PubMed

    Lang, Elisabeth; Pozdeev, Vitaly I; Xu, Haifeng C; Shinde, Prashant V; Behnke, Kristina; Hamdam, Junnat M; Lehnert, Erik; Scharf, Rüdiger E; Lang, Florian; Häussinger, Dieter; Lang, Karl S; Lang, Philipp A

    2016-01-01

    Similar to apoptosis of nucleated cells, red blood cells (RBC) can undergo suicidal cell death - called eryptosis. It is characterized by cell shrinkage and phosphatidylserine translocation. Eryptosis is triggered by an increase of intracellular calcium concentration due to activation of nonselective cation channels. The cation channels and consequently eryptosis are inhibited by erythropoietin. Eryptotic RBC are engulfed by macrophages and thus rapidly cleared from circulating blood. In this study, we explored whether storage of RBC influences the rate of eryptosis. Flow cytometry was employed to quantify phosphatidylserine exposing erythrocytes from annexin V binding and cytosolic Ca2+ activity from Fluo-3 fluorescence. Clearance of stored murine RBC was tested by injection of carboxyfluorescein succinimidyl ester (CFSE)-labelled erythrocytes. Storage for 42 days significantly increased the percentage of phosphatidylserine exposing and haemolytic erythrocytes, an effect blunted by removal of extracellular calcium. Phosphatidylserine exposure could be inhibited by addition of erythropoietin. Upon transfusion, the clearance of murine CFSE-labelled RBC from circulating blood was significantly higher following storage for 10 days when compared to 2 days of storage. Storage of RBC triggers eryptosis by Ca2+ and erythropoietin sensitive mechanisms. © 2016 The Author(s) Published by S. Karger AG, Basel.

  12. WBC count

    MedlinePlus

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

  13. Conjugated bilirubin triggers anemia by inducing erythrocyte death.

    PubMed

    Lang, Elisabeth; Gatidis, Sergios; Freise, Noemi F; Bock, Hans; Kubitz, Ralf; Lauermann, Christian; Orth, Hans Martin; Klindt, Caroline; Schuier, Maximilian; Keitel, Verena; Reich, Maria; Liu, Guilai; Schmidt, Sebastian; Xu, Haifeng C; Qadri, Syed M; Herebian, Diran; Pandyra, Aleksandra A; Mayatepek, Ertan; Gulbins, Erich; Lang, Florian; Häussinger, Dieter; Lang, Karl S; Föller, Michael; Lang, Philipp A

    2015-01-01

    Hepatic failure is commonly associated with anemia, which may result from gastrointestinal bleeding, vitamin deficiency, or liver-damaging diseases, such as infection and alcohol intoxication. At least in theory, anemia during hepatic failure may result from accelerated clearance of circulating erythrocytes. Here we show that bile duct ligation (BDL) in mice leads to severe anemia despite increased reticulocyte numbers. Bilirubin stimulated suicidal death of human erythrocytes. Mechanistically, bilirubin triggered rapid Ca(2+) influx, sphingomyelinase activation, formation of ceramide, and subsequent translocation of phosphatidylserine to the erythrocyte surface. Consistent with our in vitro and in vivo findings, incubation of erythrocytes in serum from patients with liver disease induced suicidal death of erythrocytes in relation to their plasma bilirubin concentration. Consistently, patients with hyperbilirubinemia had significantly lower erythrocyte and significantly higher reticulocyte counts compared to patients with low bilirubin levels. Bilirubin triggers suicidal erythrocyte death, thus contributing to anemia during liver disease. © 2014 The Authors. Hepatology published by Wiley Periodicals, Inc., on behalf of the American Association for the Study of Liver Diseases.

  14. Erythrocyte mechanics and blood flow

    SciTech Connect

    Cokelet, G.R.; Meiselman, H.J.; Brooks, D.E.

    1980-01-01

    This monograph includes the proceedings of a conference on erythrocyte mechanics and blood flow. The topics discussed include: the bilayer and shell model of the erythrocyte membrane; protein-protein interactions in red cell membranes; mechano-chemical study of red cell membrane structure in situ; viscoelastic solid behavior of red cell membrane; measures of blood rheology and erythrocyte mechanics; mechanisms of erythrocyte aggregation; dynamics of red blood cell deformation and aggregation, and in vivo flow; physical and mathematical models of blood flow - theoretical analysis; physical and mathematical models of blood flow - experimental studies; behavior or abnormal erythrocytes in capillaries; reduced erythrocyte deformability and vascular pathology; and microvascular transit of normal, immature, and altered red blood cells in spleen versus skeletal muscle. Summary remarks on in vitro erythrocyte characteristics and in vivo erythrocyte behavior are also indcluded. (RJC)

  15. The size of erythrocyte ghosts.

    PubMed

    Tatsumi, N

    1981-02-20

    The volume of resealed erythrocyte ghosts formed during hypotonic hemolysis of normal human erythrocytes was measured by means of a continuous mean corpuscular volume analyzer. The final volume of resealed ghosts was 140.6 +/- 15.2 fl. Strong correlations exist between the volume of ghosts and the initial mean corpuscular volume and mean corpuscular hemoglobin of the erythrocyte, and between the enlargement ratio and the mean corpuscular volume or mean corpuscular hemoglobin of the erythrocyte.

  16. Segmentation of leukocytes and erythrocytes in blood smear images.

    PubMed

    Bergen, Tobias; Steckhan, Dirk; Wittenberg, Thomas; Zerfass, Thorsten

    2008-01-01

    Differential blood count is a standard method in hematological laboratory diagnosis. In the course of developing a computer-assisted microscopy system for the generation of differential blood counts, the detection and segmentation of white and red blood cells forms an essential step and its exactness is a fundamental prerequisite for the effectiveness of the subsequent classification step. We propose a method for the exact segmentation of leukocytes and erythrocytes in a simultaneous and cooperative way. We combine pixel-wise classification with template matching to locate erythrocytes and use a level-set approach in order to get the exact cell contours of leukocyte nucleus and plasma regions as well as erythrocyte regions. An evaluation comparing the performance of the algorithm to the manual segmentation performed by several persons yielded good results.

  17. Oxidative Hemolysis of Erythrocytes

    ERIC Educational Resources Information Center

    Wlodek, Lidia; Kusior, Dorota

    2006-01-01

    This exercise for students will allow them to simultaneously observe lipid peroxidation and consequent hemolysis of rat erythrocytes and the effect of sodium azide, a catalase inhibitor, on these processes. It will also demonstrate a protective action of antioxidants, the therapeutically used N-acetylcysteine and albumins present in plasma.

  18. Oxidative Hemolysis of Erythrocytes

    ERIC Educational Resources Information Center

    Wlodek, Lidia; Kusior, Dorota

    2006-01-01

    This exercise for students will allow them to simultaneously observe lipid peroxidation and consequent hemolysis of rat erythrocytes and the effect of sodium azide, a catalase inhibitor, on these processes. It will also demonstrate a protective action of antioxidants, the therapeutically used N-acetylcysteine and albumins present in plasma.

  19. Effect of dietary zinc deficiency on the endogenous phosphorylation and dephosphorylation of rat erythrocyte membrane

    SciTech Connect

    Paterson, P.G.; Allen, O.B.; Bettger, W.J.

    1987-12-01

    The effect of dietary zinc deficiency on patterns of phosphorylation and dephosphorylation of rat erythrocyte membrane proteins and erythrocyte filterability was examined. Weanling male Wistar rats were fed an egg white-based diet containing less than 1.1 mg zinc/kg diet ad libitum for 3 wk. Control rats were either pair-fed or ad libitum-fed the basal diet supplemented with 100 mg zinc/kg diet. Net phosphorylation and dephosphorylation of erythrocyte membrane proteins were carried out by an in vitro assay utilizing (gamma-/sup 32/P)ATP. The membrane proteins were subsequently separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and the /sup 32/P content of gel slices was counted by Cerenkov counting. Erythrocyte filterability was measured as the filtration time of suspensions of erythrocytes, both untreated and preincubated with diamide, under constant pressure. Erythrocyte ghosts from zinc-deficient rats demonstrated greater dephosphorylation of protein bands R1 plus R2 and R7 than pair-fed rats and greater net phosphorylation of band R2.2 than pair-fed or ad libitum-fed control rats (P less than 0.05). Erythrocytes from ad libitum-fed control rats showed significantly longer filtration times than those from zinc-deficient or pair-fed control rats. In conclusion, dietary zinc deficiency alters in vitro patterns of erythrocyte membrane protein phosphorylation and dephosphorylation, whereas the depression in food intake associated with the zinc deficiency increases erythrocyte filterability. 71 references.

  20. The effects of superoxide dismutase knockout on the oxidative stress parameters and survival of mouse erythrocytes.

    PubMed

    Grzelak, Agnieszka; Kruszewski, Marcin; Macierzyńska, Ewa; Piotrowski, Łukasz; Pułaski, Łukasz; Rychlik, Błazej; Bartosz, Grzegorz

    2009-01-01

    The erythrocytes of 12-month old Sod1 (-/-) mice showed an increased level of reactive oxygen species (ROS), as estimated by the degree of dihydroethidine and dihydrorhodamine oxidation, and the increased level of Heinz bodies. No indices of severe oxidative stress were found in the red blood cells and blood plasma of Sod1 (-/-) mice as judged from the lack of significant changes in the levels of erythrocyte and plasma glutathione, plasma protein thiol and carbonyl groups and thiobarbituric-acid reactive substances in the blood plasma. However, a decreased erythrocyte lifespan, increased reticulocyte count and splenomegaly were noted, indicating the importance of superoxide dismutase for maintaining erythrocyte viability. The levels of erythrocyte ROS and Heinz bodies and the reticulocyte count were indistinguishable in Sod1 (+/+) and Sod1 (+/-) mice, suggesting that a superoxide dismutase activity decrease to half of its normal value may be sufficient to secure the protective effects of the enzyme.

  1. Hepatic confinement of newly produced erythrocytes caused by low-temperature exposure in Xenopus laevis.

    PubMed

    Maekawa, Shun; Iemura, Hitomi; Kuramochi, Yuko; Nogawa-Kosaka, Nami; Nishikawa, Hironori; Okui, Takehito; Aizawa, Youichi; Kato, Takashi

    2012-09-01

    Diminished erythrocyte count and erythropoiesis have been reported during hypothermia in some ectothermic animals. In this study, the African clawed frog, Xenopus laevis, was used to investigate the cause of hypothermia-induced anemia. We developed a new model of hypothermia at 5°C and monitored blood cell count and erythropoiesis on several days. Erythrocyte count declined by 30% on the first day following cold exposure (5°C) and mRNA expression of hemeoxygenase-1 was enhanced 10-fold; accumulation of iron as a result of heme degradation was observed in the liver. One day after low-temperature exposure, erythropoietin mRNA expression was elevated in the liver and lung compared with that at normal temperature (22°C) by qRT-PCR analysis. Examination of liver sections (i.e. the erythropoietic organ) showed an increase in o-dianisidine-positive erythrocytes in the hepatic sinusoid 5 days after the onset of low-temperature exposure compared with normal liver. Peripheral erythrocyte count remained low, indicating that newly produced erythrocytes did not migrate from the liver to the circulation during hypothermia. In conclusion, this study reveals hypothermic anemia as being associated with hepatic erythrocyte destruction; prolonged anemia during low-temperature exposure is concomitant with newly produced erythrocytes being confined to the liver and may lead to new insights into vertebrate hematopoiesis.

  2. Flow behavior of erythrocytes in microvessels and glass capillaries: effects of erythrocyte deformation and erythrocyte aggregation.

    PubMed

    Suzuki, Y; Tateishi, N; Soutani, M; Maeda, N

    1996-01-01

    Flow behavior of erythrocytes in microvessels and glass capillaries with an inner diameter of 10-50 microns was compared in relation to erythrocyte deformation and erythrocyte aggregation. This study was focused on the formation of a marginal cell-free layer, and the thickness was determined using an image processor. Human erythrocytes were perfused through a part of microvascular networks isolated from rabbit mesentery and through glass capillaries. Erythrocyte deformability was modified by treating erythrocytes with diamide, diazene-dicarboxylic acid bis[N,N-dimethylamide], and erythrocyte aggregation was accelerated by adding dextran (with a molecular weight of 70,400) to the perfusion medium. The thickness of the cell-free layer increased with an increase of the inner diameter of flow channel, with lowering the hematocrit, and with increasing the flow velocity of erythrocytes, in both microvessels and glass capillaries. Furthermore, the thickness of cell-free layer decreased with decreasing erythrocyte deformability, while it increased with accelerating erythrocyte aggregation. However, the alteration of the cell-free layer in response to the changes of these hemorheological conditions was more sensitive in microvessels than in glass capillaries. The present study concludes that flow behavior of erythrocytes in microvessels is qualitatively similar to, but quantitatively different from those in glass capillaries, as far as evaluated by the change of the thickness of the marginal cell-free layer.

  3. 3D visualization and quantitative analysis of human erythrocyte phagocytosis.

    PubMed

    Stachurska, Anna; Król, Teodora; Trybus, Wojciech; Szary, Karol; Fabijańska-Mitek, Jadwiga

    2016-11-01

    Since the erythrophagocytosis of opsonized erythrocytes is investigated mainly by calculating the phagocytic index using subjective light microscopy evaluation, we present methods for the quantitative and qualitative analysis of human cell erythrophagocytosis. Erythrocytes from two storage periods were used. Using Imaris software, we were able to create a three-dimensional model of erythrophagocytosis. The use of microscopy instead of cytometry revealed a significantly higher number of monocytes and erythrocytes that appeared active in phagocytosis. Spatial reconstruction allowed for detailed analysis of the process by precisely locating erythrocytes in phagocytes. Additionally, a technique of sequential image registration using Nis Elements software allowed for observation of the course of phagocytosis over a range of time intervals. This in vitro research may be helpful for understanding the cellular interactions between monocytes and erythrocytes. The cytometric method-being relatively rapid, sensitive, and specific-can serve as an alternative technique to microscopy in the quantitative analysis of erythrophagocytosis. This allows us to avoid counting the erythrocytes nonspecifically attached to monocytes and gives objective results. © 2016 International Federation for Cell Biology.

  4. O(2) release from erythrocytes flowing in a narrow O(2)-permeable tube: effects of erythrocyte aggregation.

    PubMed

    Tateishi, N; Suzuki, Y; Cicha, I; Maeda, N

    2001-07-01

    The effects of erythrocyte aggregation on O(2) release were examined using O(2)-permeable fluorinated ethylenepropylene copolymer tubes (inner diameter, 25 microm; outer diameter, 100 microm). Measurements were performed using an apparatus built on an inverted microscope that contained a scanning-grating spectrophotometer with a photon count detector connected to two photomultipliers and an image processor through a video camera. The rate of O(2) release from the cells flowing in the narrow tube was determined based on the visible absorption spectrum and the flow velocity of the cells as well as the tube size. When the tube was exposed to nitrogen-saturated deoxygenated saline containing 10 mM sodium dithionite, the flowing erythrocytes were deoxygenated in proportion to the traveling distance, and the deoxygenation at a given distance increased with decreasing flow velocity and cell concentration (hematocrit). Adding Dextran T-70 to the cell suspension increased erythrocyte aggregation in the tube, which resulted in suppressed cell deoxygenation and increased marginal cell-free-layer thickness. The deoxygenation was inversely proportional to the cell-free-layer thickness. The relation was not essentially altered even when the medium viscosity was adjusted with Dextran T-40 to remain constant. The rate of O(2) release from erythrocytes in the tube was discussed in relation to the O(2) diffusion process. We conclude that the diffusion of O(2) from erythrocytes flowing in narrow tubes is inhibited primarily by erythrocyte aggregation itself and partly by thickening of the cell-free layer.

  5. Involvement of erythrocyte aggregation and erythrocyte resistance to flow in acute coronary syndromes.

    PubMed

    Pfafferott, C; Moessmer, G; Ehrly, A M; Bauersachs, R M

    1999-01-01

    The objective of the study was to identify the relative importance of erythrocyte flow resistance and aggregation in acute and chronic coronary syndromes. 117 subjects in five groups were studied: (1) 34 patients shortly after acute myocardial infarction (AMI) before reperfusion therapy; (2) 27 patients with unstable and (3) 21 with stable angina pectoris (AP); (4) 14 age-matched control patients and (5) 21 healthy volunteers. Single erythrocyte transit times were measured using the Cell Transit Analyser. Shear dependent elongation and aggregation was measured by a modified computerized Myrenne aggregometer. Leukocyte count was increased in coronary artery disease (CAD), especially in acute syndromes (mean +/- SD for groups 1-5): 12.2 +/- 4.5; 10.0 +/- 5.4; 8.0 +/- 2.0; 8.0 +/- 3.7; 7.0 +/- 2.0 (pl(-1))). Platelets, hematocrit, fibrinogen, alpha2-macroglobulin did not differ between the groups. Plasma viscosity (mPas) was elevated in AMI and stable AP: 1.34 +/- 0.10; 1.30 +/- 0.09; 1.32 +/- 0.08; 1.27 +/- 0.07; 1.27 +/- 0.05. Erythrocyte filtrability was not different as was the shear dependent deformation. Aggregation parameters such as gammaTmin were elevated in CAD: 180 +/- 70; 159 +/- 60; 166 +/- 59; 115 +/- 43; 113 +/- 51 (s(-1)). Erythrocyte deformability, measured with two independent methods, does not appear to contribute to the pathophysiology of acute coronary syndromes. Erythrocyte aggregation and plasma viscosity were again found increased both in unstable and stable coronary disease. It is unlikely that increased red cell aggregation contributes to emergence of AMI.

  6. Cytotoxic effect of poly-dispersed single walled carbon nanotubes on erythrocytes in vitro and in vivo.

    PubMed

    Sachar, Sumedha; Saxena, Rajiv K

    2011-01-01

    Single wall Carbon Nanotubes (SWCNTs) are hydrophobic and do not disperse in aqueous solvents. Acid functionalization of SWCNTs results in attachment of carboxy and sulfonate groups to carbon atoms and the resulting acid functionalized product (AF-SWCNTs) is negatively charged and disperses easily in water and buffers. In the present study, effect of AF-SWCNTs on blood erythrocytes was examined. Incubation of mouse erythrocytes with AF-SWCNTs and not with control SWCNTs, resulted in a dose and time dependent lysis of erythrocyte. Using fluorescence tagged AF-SWCNTs, binding of AF-SWCNTs with erythrocytes could be demonstrated. Confocal microscopy results indicated that AF-SWCNTs could enter the erythrocytes. Treatment with AF-SWCNTs resulted in exposure of hydrophobic patches on erythrocyte membrane that is indicative of membrane damage. A time and dose dependent increase in externalization of phosphatidylserine on erythrocyte membrane bilayer was also found. Administration of AF-SWCNTs through intravenous route resulted in a transient anemia as seen by a sharp decline in blood erythrocyte count accompanied with a significant drop in blood haemoglobin level. Administration of AF-SWCNTs through intratracheal administration also showed significant decline in RBC count while administration through other routes (gavage and intra-peritoneal) was not effective. By using a recently developed technique of a two step in vivo biotinylation of erythrocytes that enables simultaneous enumeration of young (age <10 days) and old (age>40 days) erythrocytes in mouse blood, it was found that the in vivo toxic effect of AF-SWCNTs was more pronounced on older subpopulation of erythrocytes. Subpopulation of old erythrocytes fell after treatment with AF-SWCNTs but recovered by third day after the intravenous administration of AF-SWCNTs. Taken together our results indicate that treatment with AF-SWCNTs results in acute membrane damage and eventual lysis of erythrocytes. Intravenous

  7. Multiplicity Counting

    SciTech Connect

    Geist, William H.

    2015-12-01

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

  8. Reticulocyte count

    MedlinePlus

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

  9. Dielectric inspection of erythrocyte morphology

    NASA Astrophysics Data System (ADS)

    Hayashi, Yoshihito; Oshige, Ikuya; Katsumoto, Yoichi; Omori, Shinji; Yasuda, Akio; Asami, Koji

    2008-05-01

    We performed a systematic study of the sensitivity of dielectric spectroscopy to erythrocyte morphology. Namely, rabbit erythrocytes of four different shapes were prepared by precisely controlling the pH of the suspending medium, and their complex permittivities over the frequency range from 0.1 to 110 MHz were measured and analyzed. Their quantitative analysis shows that the characteristic frequency and the broadening parameter of the dielectric relaxation of interfacial polarization are highly specific to the erythrocyte shape, while they are insensitive to the cell volume fraction. Therefore, these two dielectric parameters can be used to differentiate erythrocytes of different shapes, if dielectric spectroscopy is applied to flow-cytometric inspection of single blood cells. In addition, we revealed the applicability and limitations of the analytical theory of interfacial polarization to explain the experimental permittivities of non-spherical erythrocytes.

  10. Influence of iodine-containing radiographic contrast media on the phenotype of erythrocytes from different laboratory animal species.

    PubMed

    Hiebl, B; Hopperdietzel, C; Hünigen, H; Dietze, K; Klein, S; Schreier, B; Jung, F

    2013-01-01

    It is well known that clinically relevant concentrations of iodine-containing radiographic contrast media (CM) induce morphological changes in human erythrocytes. However, there are only few reports about CM effects on erythrocytes of animals (e.g. mice, rats, rabbits, and pigs). Thus, two conventional iodine-containing CM (iodixanol, Visipaque™ 320; iomeprol, Iomeprol™ 350) were tested for their effects on the morphology of erythrocytes from these. After venous blood sampling and blood centrifugation, the autologous plasma was supplemented with 40 vol% CM. Then, a defined number of erythrocytes was incubated in this CM-supplemented plasma for 5 min at body temperature (37°C). Subsequently, 10 μL of the cell suspension were transferred to a purified glass slide and the number of discocytes, echinocytes, and acanthocytes was counted within a total number of 100 erythrocytes (40 fold primary magnification, transmitted light mode). Shape changes of the erythrocytes from all animal species strongly depended on the type of CM and compared to the effects which have already been described for human erythrocytes. Incubation in both CM resulted in morphological changes of the erythrocytes. Incubation in a iodixanol/plasma mixture induced the lowest echinocyte or acanthocyte formation. Porcine erythrocytes showed a much more distinct shape change than those of other animal species and humans. These results suggest erythrocytes from mice, rats, and rabbits are a suitable model system for a model system for human erythrocytes when CM effects on the cellular shape of erythrocytes have to be tested. The distinct deformation of the pig erythrocytes could be due to differences in the pig erythrocyte membrane or the physical and chemical constitution of pig erythrocytes.

  11. Tower counts

    USGS Publications Warehouse

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

    2007-01-01

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

  12. Disorders of erythrocyte volume homeostasis.

    PubMed

    Glogowska, E; Gallagher, P G

    2015-05-01

    Inherited disorders of erythrocyte volume homeostasis are a heterogeneous group of rare disorders with phenotypes ranging from dehydrated to overhydrated erythrocytes. Clinical, laboratory, physiologic, and genetic heterogeneities characterize this group of disorders. A series of recent reports have provided novel insights into our understanding of the genetic bases underlying some of these disorders of red cell volume regulation. This report reviews this progress in understanding determinants that influence erythrocyte hydration and how they have yielded a better understanding of the pathways that influence cellular water and solute homeostasis. © 2015 John Wiley & Sons Ltd.

  13. Disorders of Erythrocyte Volume Homeostasis

    PubMed Central

    Glogowska, Edyta; Gallagher, Patrick G.

    2015-01-01

    Inherited disorders of erythrocyte volume homeostasis are a heterogeneous group of rare disorders with phenotypes ranging from dehydrated to overhydrated erythrocytes. Clinical, laboratory, physiologic, and genetic heterogeneity characterize this group of disorders. A series of recent reports have provided novel insights into our understanding of the genetic bases underlying some of these disorders of red cell volume regulation. This report reviews this progress in understanding determinants that influence erythrocyte hydration and how they have yielded a better understanding of the pathways that influence cellular water and solute homeostasis. PMID:25976965

  14. White Blood Cell Count

    MedlinePlus

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

  15. Accelerated apoptotic death and in vivo turnover of erythrocytes in mice lacking functional mitogen- and stress-activated kinase MSK1/2

    PubMed Central

    Lang, Elisabeth; Bissinger, Rosi; Fajol, Abul; Salker, Madhuri S.; Singh, Yogesh; Zelenak, Christine; Ghashghaeinia, Mehrdad; Gu, Shuchen; Jilani, Kashif; Lupescu, Adrian; Reyskens, Kathleen M. S. E.; Ackermann, Teresa F.; Föller, Michael; Schleicher, Erwin; Sheffield, William P.; Arthur, J. Simon C.; Lang, Florian; Qadri, Syed M.

    2015-01-01

    The mitogen- and stress-activated kinase MSK1/2 plays a decisive role in apoptosis. In analogy to apoptosis of nucleated cells, suicidal erythrocyte death called eryptosis is characterized by cell shrinkage and cell membrane scrambling leading to phosphatidylserine (PS) externalization. Here, we explored whether MSK1/2 participates in the regulation of eryptosis. To this end, erythrocytes were isolated from mice lacking functional MSK1/2 (msk−/−) and corresponding wild-type mice (msk+/+). Blood count, hematocrit, hemoglobin concentration and mean erythrocyte volume were similar in both msk−/− and msk+/+ mice, but reticulocyte count was significantly increased in msk−/− mice. Cell membrane PS exposure was similar in untreated msk−/− and msk+/+ erythrocytes, but was enhanced by pathophysiological cell stressors ex vivo such as hyperosmotic shock or energy depletion to significantly higher levels in msk−/− erythrocytes than in msk+/+ erythrocytes. Cell shrinkage following hyperosmotic shock and energy depletion, as well as hemolysis following decrease of extracellular osmolarity was more pronounced in msk−/− erythrocytes. The in vivo clearance of autologously-infused CFSE-labeled erythrocytes from circulating blood was faster in msk−/− mice. The spleens from msk−/− mice contained a significantly greater number of PS-exposing erythrocytes than spleens from msk+/+ mice. The present observations point to accelerated eryptosis and subsequent clearance of erythrocytes leading to enhanced erythrocyte turnover in MSK1/2-deficient mice. PMID:26611568

  16. Accelerated apoptotic death and in vivo turnover of erythrocytes in mice lacking functional mitogen- and stress-activated kinase MSK1/2.

    PubMed

    Lang, Elisabeth; Bissinger, Rosi; Fajol, Abul; Salker, Madhuri S; Singh, Yogesh; Zelenak, Christine; Ghashghaeinia, Mehrdad; Gu, Shuchen; Jilani, Kashif; Lupescu, Adrian; Reyskens, Kathleen M S E; Ackermann, Teresa F; Föller, Michael; Schleicher, Erwin; Sheffield, William P; Arthur, J Simon C; Lang, Florian; Qadri, Syed M

    2015-11-27

    The mitogen- and stress-activated kinase MSK1/2 plays a decisive role in apoptosis. In analogy to apoptosis of nucleated cells, suicidal erythrocyte death called eryptosis is characterized by cell shrinkage and cell membrane scrambling leading to phosphatidylserine (PS) externalization. Here, we explored whether MSK1/2 participates in the regulation of eryptosis. To this end, erythrocytes were isolated from mice lacking functional MSK1/2 (msk(-/-)) and corresponding wild-type mice (msk(+/+)). Blood count, hematocrit, hemoglobin concentration and mean erythrocyte volume were similar in both msk(-/-) and msk(+/+) mice, but reticulocyte count was significantly increased in msk(-/-) mice. Cell membrane PS exposure was similar in untreated msk(-/-) and msk(+/+) erythrocytes, but was enhanced by pathophysiological cell stressors ex vivo such as hyperosmotic shock or energy depletion to significantly higher levels in msk(-/-) erythrocytes than in msk(+/+) erythrocytes. Cell shrinkage following hyperosmotic shock and energy depletion, as well as hemolysis following decrease of extracellular osmolarity was more pronounced in msk(-/-) erythrocytes. The in vivo clearance of autologously-infused CFSE-labeled erythrocytes from circulating blood was faster in msk(-/-) mice. The spleens from msk(-/-) mice contained a significantly greater number of PS-exposing erythrocytes than spleens from msk(+/+) mice. The present observations point to accelerated eryptosis and subsequent clearance of erythrocytes leading to enhanced erythrocyte turnover in MSK1/2-deficient mice.

  17. Counting Penguins.

    ERIC Educational Resources Information Center

    Perry, Mike; Kader, Gary

    1998-01-01

    Presents an activity on the simplification of penguin counting by employing the basic ideas and principles of sampling to teach students to understand and recognize its role in statistical claims. Emphasizes estimation, data analysis and interpretation, and central limit theorem. Includes a list of items for classroom discussion. (ASK)

  18. Counting Populations

    ERIC Educational Resources Information Center

    Damonte, Kathleen

    2004-01-01

    Scientists use sampling to get an estimate of things they cannot easily count. A population is made up of all the organisms of one species living together in one place at the same time. All of the people living together in one town are considered a population. All of the grasshoppers living in a field are a population. Scientists keep track of the…

  19. Counting Penguins.

    ERIC Educational Resources Information Center

    Perry, Mike; Kader, Gary

    1998-01-01

    Presents an activity on the simplification of penguin counting by employing the basic ideas and principles of sampling to teach students to understand and recognize its role in statistical claims. Emphasizes estimation, data analysis and interpretation, and central limit theorem. Includes a list of items for classroom discussion. (ASK)

  20. Counting Populations

    ERIC Educational Resources Information Center

    Damonte, Kathleen

    2004-01-01

    Scientists use sampling to get an estimate of things they cannot easily count. A population is made up of all the organisms of one species living together in one place at the same time. All of the people living together in one town are considered a population. All of the grasshoppers living in a field are a population. Scientists keep track of the…

  1. Migraine and erythrocyte biology: a review.

    PubMed

    Lippi, G; Cervellin, G; Mattiuzzi, C

    2014-12-01

    Migraine is a common disabling headache disorder that is conventionally classified according to the presence or absence of aura. The pathogenesis of this disorder entails a complex interplay of neurovascular factors, that trigger reduction of cerebral blood flow followed by reactive vasodilatation. Despite major emphasis has been placed on the investigation of putative biomarkers that could predict response to specific treatments and prophylaxis, less focus has been directed at the association between migraine and erythrocytosis. Erythrocytosis is typically accompanied by hyperviscosity, that is now considered a crucial determinant in the pathogenesis of migraine. The results of some epidemiological investigations are in substantial agreement to confirm the existence of a significant relationship between increased haemoglobin levels and migraine, whereas some case reports have also reported an effective improvement of symptoms after reduction of erythrocyte count by therapeutic venesection. Interesting evidence has recently emerged from the assessment of red blood cell distribution width (RDW), a simple and inexpensive measure of anysocytosis that has been also associated with a variety of ischaemic and thrombotic disorders other than migraine. The aim of this review was to provide an overview of the current clinical and epidemiological evidence linking migraine and erythrocyte biology.

  2. Abnormal erythrocyte metabolism in hepatic disease.

    PubMed

    Smith, J R; Kay, N E; Gottlieb, A J; Oski, F A

    1975-12-01

    Erythrocyte (RBC) metabolic studies were done on 114 patients with severe hepatic disease. Heinz body formation after incubation of RBCs with acetyl phenylhydrazine was found to be significantly higher in patients than in controls. RBC-reduced glutathione levels were lower than those of controls both before and after incubation with acetyl phenylhydrazine, and patients with the highest Heinz body counts had the lowest reduced glutathione levels. RBC methylene blue-stimulated hexose monophosphate (HMP) shunt metabolism and glucose recycling through the shunt were significantly lower in patients with active hepatic disease than in controls. There was no difference in resting HMP shunt activity or in resting recycling of glucose. Despite impairment of shunt metabolism, total glucose consumption was greater in patients than in controls. The patients with the lowest stimulated HMP shunt metabolism and glucose recycling had the highest Heinz body counts, lowest reduced glutathione, and highest total glucose consumption. A continuum of abnormal shunt metabolism was seen, from a mild reduction of stimulated HMP shunt activity to a severe combined decrease in both the HMP shunt and glucose recycling. When measured, glutathione reductase, glutathione peroxidase, glucose-6-phosphate dehydrogenase, and transketolase were normal or increased. Sequential studies were done on 11 patients who had abnormal metabolic studies. Coincident with improvement of HMP shunt metabolism, the Heinz body counts became lower, reduced glutathione higher, hematocrit higher, and liver function improved. Impaired HMP shunt metabolism appears to be a common, acquired RBC abnormality in patients with severe, active liver disease.

  3. Increased Microerythrocyte Count in Homozygous α+-Thalassaemia Contributes to Protection against Severe Malarial Anaemia

    PubMed Central

    Fowkes, Freya J. I; Allen, Stephen J; Allen, Angela; Alpers, Michael P; Weatherall, David J; Day, Karen P

    2008-01-01

    Background The heritable haemoglobinopathy α+-thalassaemia is caused by the reduced synthesis of α-globin chains that form part of normal adult haemoglobin (Hb). Individuals homozygous for α+-thalassaemia have microcytosis and an increased erythrocyte count. α+-Thalassaemia homozygosity confers considerable protection against severe malaria, including severe malarial anaemia (SMA) (Hb concentration < 50 g/l), but does not influence parasite count. We tested the hypothesis that the erythrocyte indices associated with α+-thalassaemia homozygosity provide a haematological benefit during acute malaria. Methods and Findings Data from children living on the north coast of Papua New Guinea who had participated in a case-control study of the protection afforded by α+-thalassaemia against severe malaria were reanalysed to assess the genotype-specific reduction in erythrocyte count and Hb levels associated with acute malarial disease. We observed a reduction in median erythrocyte count of ∼1.5 × 1012/l in all children with acute falciparum malaria relative to values in community children (p < 0.001). We developed a simple mathematical model of the linear relationship between Hb concentration and erythrocyte count. This model predicted that children homozygous for α+-thalassaemia lose less Hb than children of normal genotype for a reduction in erythrocyte count of >1.1 × 1012/l as a result of the reduced mean cell Hb in homozygous α+-thalassaemia. In addition, children homozygous for α+-thalassaemia require a 10% greater reduction in erythrocyte count than children of normal genotype (p = 0.02) for Hb concentration to fall to 50 g/l, the cutoff for SMA. We estimated that the haematological profile in children homozygous for α+-thalassaemia reduces the risk of SMA during acute malaria compared to children of normal genotype (relative risk 0.52; 95% confidence interval [CI] 0.24–1.12, p = 0.09). Conclusions The increased erythrocyte count and microcytosis in

  4. Increased microerythrocyte count in homozygous alpha(+)-thalassaemia contributes to protection against severe malarial anaemia.

    PubMed

    Fowkes, Freya J I; Allen, Stephen J; Allen, Angela; Alpers, Michael P; Weatherall, David J; Day, Karen P

    2008-03-18

    The heritable haemoglobinopathy alpha(+)-thalassaemia is caused by the reduced synthesis of alpha-globin chains that form part of normal adult haemoglobin (Hb). Individuals homozygous for alpha(+)-thalassaemia have microcytosis and an increased erythrocyte count. Alpha(+)-thalassaemia homozygosity confers considerable protection against severe malaria, including severe malarial anaemia (SMA) (Hb concentration < 50 g/l), but does not influence parasite count. We tested the hypothesis that the erythrocyte indices associated with alpha(+)-thalassaemia homozygosity provide a haematological benefit during acute malaria. Data from children living on the north coast of Papua New Guinea who had participated in a case-control study of the protection afforded by alpha(+)-thalassaemia against severe malaria were reanalysed to assess the genotype-specific reduction in erythrocyte count and Hb levels associated with acute malarial disease. We observed a reduction in median erythrocyte count of approximately 1.5 x 10(12)/l in all children with acute falciparum malaria relative to values in community children (p < 0.001). We developed a simple mathematical model of the linear relationship between Hb concentration and erythrocyte count. This model predicted that children homozygous for alpha(+)-thalassaemia lose less Hb than children of normal genotype for a reduction in erythrocyte count of >1.1 x 10(12)/l as a result of the reduced mean cell Hb in homozygous alpha(+)-thalassaemia. In addition, children homozygous for alpha(+)-thalassaemia require a 10% greater reduction in erythrocyte count than children of normal genotype (p = 0.02) for Hb concentration to fall to 50 g/l, the cutoff for SMA. We estimated that the haematological profile in children homozygous for alpha(+)-thalassaemia reduces the risk of SMA during acute malaria compared to children of normal genotype (relative risk 0.52; 95% confidence interval [CI] 0.24-1.12, p = 0.09). The increased erythrocyte count and

  5. Avian leucocyte counting using the hemocytometer

    USGS Publications Warehouse

    Dein, F.J.; Wilson, A.; Fischer, D.; Langenberg, P.

    1994-01-01

    Automated methods for counting leucocytes in avian blood are not available because of the presence of nucleated erythrocytes and thrombocytes. Therefore, total white blood cell counts are performed by hand using a hemocytometer. The Natt and Herrick and the Unopette methods are the most common stain and diluent preparations for this procedure. Replicate hemocytometer counts using these two methods were performed on blood from four birds of different species. Cells present in each square of the hemocytometer were counted. Counting cells in the corner, side, or center hemocytometer squares produced statistically equivalent results; counting four squares per chamber provided a result similar to that obtained by counting nine squares; and the Unopette method was more precise for hemocytometer counting than was the Natt and Herrick method. The Unopette method is easier to learn and perform but is an indirect process, utilizing the differential count from a stained smear. The Natt and Herrick method is a direct total count, but cell identification is more difficult.

  6. [Influence of pleural fluid red blood cell count on the misidentification of transudates].

    PubMed

    Porcel, José Manuel; Esquerda, Aureli; Martínez, Montserrat; Rodríguez-Panadero, Francisco; Bielsa, Silvia

    2008-12-06

    Light's criteria misclassify a quarter of transudates as exudates. We assessed the influence of red blood cell counts on pleural lactate dehydrogenase (LDH) levels and, thereby, on the specificity of Light's criteria. We retrospectively reviewed 1,312 consecutive patients with pleural effusion, of whom 1,014 were exudates and 298 transudates according to clinical criteria. The relationship between pleural erythrocytes and LDH using simple linear regression analysis, as well as the operating characteristics of Light's criteria, were assessed. Finally, a formula to correct pleural LDH levels, according to the erythrocyte count, was generated. There was a linear relationship between the pleural erythrocyte count and LDH levels (r = 0.44; p < 0.001). Light's criteria yielded 81% specificity in patients with pleural erythrocyte counts < or = 10.000 3 10(6)/l, as compared to 61% in a group with a higher erythrocyte counts (p < 0.01). The application of the LDH formula enabled the correct reclassification of 24 of 64 (37%) false exudates. A high pleural erythrocyte count, through its influence on the LDH levels, may lead to a transudate being misclassified as an exudate after applying Light's criteria.

  7. AMPKalpha1 deletion shortens erythrocyte life span in mice: role of oxidative stress.

    PubMed

    Wang, Shaobin; Dale, George L; Song, Ping; Viollet, Benoit; Zou, Ming-Hui

    2010-06-25

    AMP-activated protein kinase (AMPK) is an energy sensor essential for maintaining cellular energy homeostasis. Here, we report that AMPKalpha1 is the predominant isoform of AMPK in murine erythrocytes and mice globally deficient in AMPKalpha1 (AMPKalpha1(-/-)), but not in those lacking AMPKalpha2, and the mice had markedly enlarged spleens with dramatically increased proportions of Ter119-positive erythroid cells. Blood tests revealed significantly decreased erythrocyte and hemoglobin levels with increased reticulocyte counts and elevated plasma erythropoietin concentrations in AMPKalpha1(-/-) mice. The life span of erythrocytes from AMPKalpha1(-/-) mice was less than that in wild-type littermates, and the levels of reactive oxygen species and oxidized proteins were significantly increased in AMPKalpha1(-/-) erythrocytes. In keeping with the elevated oxidative stress, treatment of AMPKalpha1(-/-) mice with the antioxidant, tempol, resulted in decreased reticulocyte counts and improved erythrocyte survival. Furthermore, the expression of Foxo3 and reactive oxygen species scavenging enzymes was significantly decreased in erythroblasts from AMPKalpha1(-/-) mice. Collectively, these results establish an essential role for AMPKalpha1 in regulating oxidative stress and life span in erythrocytes.

  8. Erythrocyte nitric oxide availability and oxidative stress following exercise.

    PubMed

    Medeiros-Lima, Daniel Jose Matos; Mendes-Ribeiro, Antonio Claudio; Brunini, Tatiana Marlowe Cunha; Martins, Marcela Anjos; Mury, Wanda Vianna; Freire, Raul Almeira; Monteiro, Walace David; Farinatti, Paulo Tarso Veras; Matsuura, Cristiane

    2017-01-01

    Growing evidence has shown that acute exercise impairs erythrocyte membrane structure and function as a consequence of increased physical and chemical stress. Erythrocyte-synthesized nitric oxide (NO) is known to modulate membrane fluidity, and its bioavailability depends on the balance between its production and scavenging by reactive oxygen species. Here, we investigated whether a maximal exercise test could affect erythrocyte NO bioavailability and oxidative stress. Twelve men (26±4 years old, V̇O2peak 44.1±4.3 mL·kg-1·min-1) performed a treadmill maximal cardiopulmonary exercise test. Blood was collected at rest and immediately after exercise for erythrocytes isolation. Maximal exercise caused an increase in erythrocytes count, haemoglobin and haematocrit levels. There was no change in L-arginine influx into erythrocytes after exercise. Yet, nitric oxide synthase activity, and thus, NO production, was increased after maximal test, as well cyclic GMP levels. In relation to biomarkers of oxidative stress, maximal test resulted in increased levels of lipid peroxidation, and diminished superoxide dismutase activity. Neither glutathione peroxidase nor catalase activity was affected by maximal test. Our findings demonstrate that the increased erythrocyte membrane rigidity caused by an acute bout of exercise may be caused, in part, by an increased lipid oxidative damage caused by ROS produced exogenously.

  9. Physiological and hematological changes in Chum Salmon artificially infected with Erythrocytic Necrosis virus

    USGS Publications Warehouse

    Haney, D.C.; Hursh, D.A.; Mix, M.C.; Winton, J.R.

    1992-01-01

    Chum salmon Oncorhynchus keta were injected with erythrocytic necrosis virus (ENV) to study the physiological and hematological consequences of ENV infection. Infected and control fish were held in pathogen-free seawater and sampled for 5 weeks. Physiological tests included measures of plasma cortisol, glucose, protein, and osmolality; blood lactic acid; and liver glycogen. In general, ENV-infected fish had lower plasma glucose and blood lactic acid, and higher liver glycogen concentrations than did control fish. Hematological tests included red and white blood cell (RBC and WBC) counts, hematocrit, measurement of blood hemoglobin concentration, and a determination of erythrocyte fragility. Infected fish had lower RBC counts, hematocrits, and hemoglobin concentrations; higher WBC counts; and less fragile erythrocytes than did control fish. The hematology data indicated that erythrocytes of infected fish had higher mean corpuscular volume, depressed mean corpuscular hemoglobin concentration, and slightly lower mean corpuscular hemoglobin. Erythrocytic inclusions were observed in the cytoplasm of RBCs from infected fish. The infection progressed steadily through week 4, after which the fish appeared to begin recovering. In a second study, fish were infected with ENV for 3 weeks, and recovery from a stress challenge test was measured. Plasma glucose concentrations and osmclality were higher in infected fish, whereas plasma cortisol and blood lactate were only slightly elevated. These studies indicate that chum salmon withstood the effects of ENV infection without in-eversible physiological consequences. However, when subjected to a stress challenge test, infected fish recovered more slowly than control fish and had increased osmoregulatory difficulties.

  10. Abnormalities of the erythrocyte membrane.

    PubMed

    Gallagher, Patrick G

    2013-12-01

    Primary abnormalities of the erythrocyte membrane are characterized by clinical, laboratory, and genetic heterogeneity. Among this group, hereditary spherocytosis patients are more likely to experience symptomatic anemia. Treatment of hereditary spherocytosis with splenectomy is curative in most patients. Growing recognition of the long-term risks of splenectomy has led to re-evaluation of the role of splenectomy. Management guidelines acknowledge these considerations and recommend discussion between health care providers, patient, and family. The hereditary elliptocytosis syndromes are the most common primary disorders of erythrocyte membrane proteins. However, most elliptocytosis patients are asymptomatic and do not require therapy.

  11. Fetal-maternal erythrocyte distribution blood test

    MedlinePlus

    Kleihauer-Betke stain; Flow cytometry - fetal-maternal erythrocyte distribution; Rh incompatibility - erythrocyte distribution ... slightly among different laboratories. Some labs use different measurements or test different samples. Talk to your doctor ...

  12. [Study of erythrocyte dehydration using spin labels].

    PubMed

    Moiseev, V A; Mezhidov, S Kh; Nardid, O A

    1989-01-01

    Possibility of studying erythrocyte dehydration by ESR-spin probe is substantiated. Dehydration of erythrocytes in relation to osmolarity of sodium chloride solutions is investigated. The results are shown to agree with the data obtained by radioisotope method.

  13. Blunted apoptosis of erythrocytes in mice deficient in the heterotrimeric G-protein subunit Gαi2

    PubMed Central

    Bissinger, Rosi; Lang, Elisabeth; Ghashghaeinia, Mehrdad; Singh, Yogesh; Zelenak, Christine; Fehrenbacher, Birgit; Honisch, Sabina; Chen, Hong; Fakhri, Hajar; Umbach, Anja T.; Liu, Guilai; Rexhepaj, Rexhep; Liu, Guoxing; Schaller, Martin; Mack, Andreas F.; Lupescu, Adrian; Birnbaumer, Lutz; Lang, Florian; Qadri, Syed M.

    2016-01-01

    Putative functions of the heterotrimeric G-protein subunit Gαi2-dependent signaling include ion channel regulation, cell differentiation, proliferation and apoptosis. Erythrocytes may, similar to apoptosis of nucleated cells, undergo eryptosis, characterized by cell shrinkage and cell membrane scrambling with phosphatidylserine (PS) exposure. Eryptosis may be triggered by increased cytosolic Ca2+ activity and ceramide. In the present study, we show that Gαi2 is expressed in both murine and human erythrocytes and further examined the survival of erythrocytes drawn from Gαi2-deficient mice (Gαi2−/−) and corresponding wild-type mice (Gαi2+/+). Our data show that plasma erythropoietin levels, erythrocyte maturation markers, erythrocyte counts, hematocrit and hemoglobin concentration were similar in Gαi2−/− and Gαi2+/+ mice but the mean corpuscular volume was significantly larger in Gαi2−/− mice. Spontaneous PS exposure of circulating Gαi2−/− erythrocytes was significantly lower than that of circulating Gαi2+/+ erythrocytes. PS exposure was significantly lower in Gαi2−/− than in Gαi2+/+ erythrocytes following ex vivo exposure to hyperosmotic shock, bacterial sphingomyelinase or C6 ceramide. Erythrocyte Gαi2 deficiency further attenuated hyperosmotic shock-induced increase of cytosolic Ca2+ activity and cell shrinkage. Moreover, Gαi2−/− erythrocytes were more resistant to osmosensitive hemolysis as compared to Gαi2+/+ erythrocytes. In conclusion, Gαi2 deficiency in erythrocytes confers partial protection against suicidal cell death. PMID:27499046

  14. Indirect vs direct measurement of magnesium and zinc in erythrocytes.

    PubMed

    Deuster, P A; Trostmann, U H; Bernier, L L; Dolev, E

    1987-04-01

    We evaluated three methods (two indirect and one direct) for determining the magnesium (Mg) and zinc (Zn) content of erythrocytes, to compare methodologic differences and to establish a method suitable for use in field studies. For the indirect methods, erythrocytes in whole blood were lysed by adding either de-ionized water (I) or nitric acid, 2 mol/L (II). For the direct method (III), erythrocytes were isolated by density centrifugation, washed, then digested in concentrated HNO3. Mg and Zn concentrations were measured by atomic absorption spectrophotometry in plasma and whole blood for the indirect methods, and in the pellet for the direct method. Hematocrit and hemoglobin were measured, and erythrocytes were sized and counted on all samples. Within-run CVs for the three methods ranged from 2.2% with method III for Mg to 5.4% with method I for Zn. CVs for reproducibility of the analytical procedures ranged from 2.6% with method II for Zn to 4.2% with method I for the two cations. Analytical recoveries of added Mg and Zn ranged from 93.8 to 104.7%. When values for the three methods were compared, those by method I were significantly (p less than 0.05) lower than those by methods II and III. Values obtained by method II were 100.1% for Mg and 102.4% for Zn of those by the direct method. Thus, the indirect method with 2 mol/L HNO3 lysing solution provides a reproducible, reliable, accurate, and simple technique for measuring Mg and Zn in erythrocytes.

  15. [Seasonal changes in mechanical resistance of erythrocytes of the long-tailed ground squirrel (Citellus undulatus)].

    PubMed

    Gulevsky, A K; Shchenyavsky, I I

    2014-01-01

    Seasonal changes of relative blood reticulocyte counts, free plasma hemoglobin and mechanical erythrocyte resistance were studied in the long-tailed ground squirrel (Citellus undulatus), under different functional conditions (winter hibernation, forced arousal in winter, and summer activity). A significant increase in reticulocyte counts in the ground squirrel blood was observed in April and October, indicating enhancement of erythropoeisis. This conclusion is also confirmed by a significant increase in free plasma hemoglobin at these periods. It has been also shown that erythrocytes of hibernating and forcibly awaken winter ground squirrels have a significantly lower mechanical resistance in comparison with those of active summer animals. The obtained data indicate that during the periods preceding the onset of winter hibernation and transition to summer activity there occurs a seasonal replacement of the erythrocyte pool by a pool more adapted to performance of functions in the novel functional state of the animal-hibernator.

  16. Agglutination of Mouse Erythrocytes by Eperythrozoon coccoides

    PubMed Central

    Iralu, Vichazelhu; Ganong, Kevin D.

    1983-01-01

    Erythrocytes from blood of mice infected with Eperythrozoon coccoides for 3 or 4 days agglutinated spontaneously. Washed E. coccoides particles agglutinated washed erythrocytes of uninfected mice. E. coccoides-mediated agglutination of normal mouse erythrocytes would be an excellent system for studies of bacterial adhesion. Images PMID:6832825

  17. A Demonstration of Erythrocyte Membrane Asymmetry.

    ERIC Educational Resources Information Center

    Pederson, Philip; And Others

    1985-01-01

    A three-period experiment was developed to help students visualize asymmetric distribution of proteins within membranes. It includes: (1) isolating erythrocyte membranes; (2) differential labeling of intact erythrocytes and isolated erythrocyte membranes with an impermeable fluorescent dye; and (3) separating proteins by polyacrylamide gel…

  18. Studies on metabolically depleted erythrocytes.

    PubMed

    Reinhart, S A; Schulzki, T; Bonetti, P O; Reinhart, W H

    2014-01-01

    Erythrocytes kept outside the blood circulation undergo progressive changes in metabolism, shape and function, which was the topic of this study. For that purpose, blood anticoagulated with either heparin, citrate or EDTA was incubated at temperatures of 5°C, 22°C or 37°C for 0 h, 24 h and 48 h, respectively. A temperature- and time-dependent decrease of glucose and ATP and increase of lactate and LDH were observed. An erythrocyte swelling and echinocytic shape transformation, which was also time- and temperature-dependent, was seen. Density-separated young and old erythrocytes behaved similarly. The degree of echinocytic shape transformation correlated with the increase in blood viscosity at high shear rate. Echinocytosis was partially reversible when erythrocytes were suspended in buffer containing 0.2% albumin. This phenomenon is specific for albumin, since molecules with a similar molecular weight (dextran 70, heat shock protein, protein C) had no effect. These finding may have an impact on blood banking and transfusion medicine.

  19. Altered erythropoiesis and decreased number of erythrocytes in children with neuroblastoma

    PubMed Central

    Morandi, Fabio; Barco, Sebastiano; Stigliani, Sara; Croce, Michela; Persico, Luca; Lagazio, Corrado; Scuderi, Francesca; Belli, Maria Luisa; Montera, Mariapina; Cangemi, Giuliana; Pozzi, Sarah; Rigo, Valentina; Scaruffi, Paola; Amoroso, Loredana; Erminio, Giovanni; Pistoia, Vito; Ferrini, Silvano; Corrias, Maria Valeria

    2017-01-01

    Neuroblastoma (NB) is a pediatric tumor presenting at diagnosis either as localized or metastatic disease, which mainly involves the bone marrow (BM). The physical occupancy of BM space by metastatic NB cells has been held responsible for impairment of BM function. Here, we investigated whether localized or metastatic NB may alter hematopoietic lineages’ maturation and release of mature cells in the periphery, through gene expression profiling, analysis of BM smears, cell blood count and flow cytometry analysis. Gene ontology and disease-associated analysis of the genes significantly under-expressed in BM resident cells from children with localized and metastatic NB, as compared to healthy children, indicated anemia, blood group antigens, and heme and porphyrin biosynthesis as major functional annotation clusters. Accordingly, in children with NB there was a selective impairment of erythrocyte maturation at the ortho-chromic stage that resulted in reduced erythrocyte count in the periphery, regardless of the presence of metastatic cells in the BM. By considering all NB patients, low erythrocyte count at diagnosis associated with worse survival. Moreover, in the subset of metastatic patients, low erythrocyte count, hemoglobin and hematocrit and high red cell distribution width at follow-up also associated with worse outcome. These observations provide an alternative model to the tenet that infiltrating cells inhibit BM functions due to physical occupancy of space and may open a new area of research in NB to understand the mechanism(s) responsible for such selective impairment. PMID:28881804

  20. Relationship between serum cholesterol and indices of erythrocytes and platelets in the US population[S

    PubMed Central

    Fessler, Michael B.; Rose, Kathryn; Zhang, Yanmei; Jaramillo, Renee; Zeldin, Darryl C.

    2013-01-01

    Whereas dyslipidemia has been associated with leukocytosis, the relationship between serum cholesterol and other hematopoietic lineages is poorly defined. Erythrocytes and platelets, anucleate cells relegated to nonspecific diffusional exchange of cholesterol with serum, have been proposed to have a distinct relationship to cholesterol from leukocytes. We examined the relationship between serum cholesterol and circulating erythrocyte/platelet indices in 4,469 adult participants of the National Health and Nutrition Examination Survey (NHANES) 2005–2006. In linear regression analyses, serum non-high density lipoprotein-cholesterol (non-HDL-C) was positively associated with mean erythrocyte number, hematocrit, hemoglobin concentration, platelet count, and platelet crit independently of age, gender, race/ethnicity, smoking, body mass index, serum folate, and C-reactive protein. The magnitude of the relationship was most marked for platelets, with lowest versus highest non-HDL-C quartile subjects having geometric mean platelet counts of 258,000/μl versus 281,000/μl, respectively (adjusted model, P < 0.001 for trend). These associations persisted in a sensitivity analysis excluding several conditions that affect erythrocyte/platelet and/or serum cholesterol levels, and were also noted in an independent analysis of 5,318 participants from NHANES 2007–2008. As non-HDL-C, erythrocytes, and platelets all impact cardiovascular disease risk, there is a need for advancing understanding of the underlying interactions that govern levels of these three blood components. PMID:23999863

  1. Relationship between serum cholesterol and indices of erythrocytes and platelets in the US population.

    PubMed

    Fessler, Michael B; Rose, Kathryn; Zhang, Yanmei; Jaramillo, Renee; Zeldin, Darryl C

    2013-11-01

    Whereas dyslipidemia has been associated with leukocytosis, the relationship between serum cholesterol and other hematopoietic lineages is poorly defined. Erythrocytes and platelets, anucleate cells relegated to nonspecific diffusional exchange of cholesterol with serum, have been proposed to have a distinct relationship to cholesterol from leukocytes. We examined the relationship between serum cholesterol and circulating erythrocyte/platelet indices in 4,469 adult participants of the National Health and Nutrition Examination Survey (NHANES) 2005-2006. In linear regression analyses, serum non-high density lipoprotein-cholesterol (non-HDL-C) was positively associated with mean erythrocyte number, hematocrit, hemoglobin concentration, platelet count, and platelet crit independently of age, gender, race/ethnicity, smoking, body mass index, serum folate, and C-reactive protein. The magnitude of the relationship was most marked for platelets, with lowest versus highest non-HDL-C quartile subjects having geometric mean platelet counts of 258,000/μl versus 281,000/μl, respectively (adjusted model, P < 0.001 for trend). These associations persisted in a sensitivity analysis excluding several conditions that affect erythrocyte/platelet and/or serum cholesterol levels, and were also noted in an independent analysis of 5,318 participants from NHANES 2007-2008. As non-HDL-C, erythrocytes, and platelets all impact cardiovascular disease risk, there is a need for advancing understanding of the underlying interactions that govern levels of these three blood components.

  2. Exploring fish bioassay of textile dye wastewaters and their selected constituents in terms of mortality and erythrocyte disorders.

    PubMed

    Sharma, Shweta; Sharma, Subhasini; Singh, P K; Swami, R C; Sharma, K P

    2009-07-01

    Acute (4 day) and short-term (7 day) toxicity studies (at 1/5th and 1/10th of LC(50)) of textile dye wastewaters and their selected ingredients (azo dye methyl red and heavy metals Cd, Cu, Ni and Zn) were made on a freshwater fish Gambusia affinis under laboratory conditions. LC(50) value was found to be the lowest in four cases, and the EC(50) value for reduction in erythrocyte counts in the remaining four tests. Thus, the reduction in erythrocyte counts to the 50% level was similar in sensitivity to fish mortality. The short-term toxicity studies revealed significant disorders in erythrocyte morphology (poikilocytosis) and its counts to be the better indices for toxicity monitoring in the absence of fish mortality.

  3. Diet of Racing Sled Dogs Affects Erythrocyte Depression by Stress

    PubMed Central

    Adkins, T. O.; Kronfeld, D. S.

    1982-01-01

    Fourteen racing huskies were matched into pairs then assigned to two diets, a commercial stress diet and an experimental diet. Proportions of protein: fat:carbohydrate on an available energy basis were 23:57:20 in a commercial stress diet and 28:69:3 in an experimental diet. The team participated in the 1979 Iditarod Trail race and was overtaken by an episode of diarrhea. Clinical signs were suggestive of parvovirus infection; high serum titers of parvo antibodies were found after the race. Blood examination showed normal levels of metabolites, electrolytes and enzymes after the race. Erythrocyte counts were depressed significantly during the race, by 15% in dogs fed an experimental diet and by 27% in those fed a commercial stress diet. Erythrocyte parameters have also become depressed during the racing season in middle distance sled dogs fed 28% protein (energy basis) but not 32 or 39%. Depressed red blood cell production has been demonstrated previously in dogs subjected to stress induced experimentally in several ways, and its restoration has been affected by dietary protein. Erythrocyte parameters may be useful indicies of the degree of stress in a dog as well as the adequacy of its protein intake during stress. PMID:17422178

  4. Tropomyosin modulates erythrocyte membrane stability

    PubMed Central

    An, Xiuli; Salomao, Marcela; Guo, Xinhua; Gratzer, Walter; Mohandas, Narla

    2007-01-01

    The ternary complex of spectrin, actin, and 4.1R (human erythrocyte protein 4.1) defines the nodes of the erythrocyte membrane skeletal network and is inseparable from membrane stability under mechanical stress. These junctions also contain tropomyosin (TM) and the other actin-binding proteins, adducin, protein 4.9, tropomodulin, and a small proportion of capZ, the functions of which are poorly defined. Here, we have examined the consequences of selective elimination of TM from the membrane. We have shown that the mechanical stability of the membranes of resealed ghosts devoid of TM is grossly, but reversibly, impaired. That the decreased membrane stability of TM-depleted membranes is the result of destabilization of the ternary complex of the network junctions is demonstrated by the strongly facilitated entry into the junctions in situ of a β-spectrin peptide, containing the actin- and 4.1R-binding sites, after extraction of the TM. The stabilizing effect of TM is highly specific, in that it is only the endogenous isotype, and not the slightly longer muscle TM that can bind to the depleted membranes and restore their mechanical stability. These findings have enabled us identify a function for TM in elevating the mechanical stability of erythrocyte membranes by stabilizing the spectrin-actin-4.1R junctional complex. PMID:17008534

  5. Triton shells of intact erythrocytes.

    PubMed

    Sheetz, M P; Sawyer, D

    1978-01-01

    About 40% of human erythrocyte membrane protein is resistant to solubilization in 0.5% Triton X-114. These components comprise a structure called a Triton shell roughly similar in size and shape to the original erythrocyte and thus constitute a cytoskeleton. With increasing concentrations of Triton the lipid content of the Triton shell decreases dramatically, whereas the majority of the protein components remain constant. Exceptions to this rule include proteins contained in band 3, the presumed anion channel, and in band 4 which decrease with increasing Triton concentration. The Triton-insoluble complex includes spectrin (bands 1 and 2), actin (band 5), and bands 3' and 7. Component 3' has an apparent molecular weight of 88,000 daltons as does 3; but unlike 3, it is insensitive to protease treatment of the intact cell, has a low extinction coefficient at 280 nm, and is solubilized from the shells in alkaline water solutions. Component 7 also has a low extinction coefficient at 280 nm. Spectrin alone is solubilized from the Triton shells in isotonic media. The solubilized spectrin contains no bound Triton and coelectrophoreses with spectrin eluted in hypotonic solutions from ghosts. Electron micrographs of fixed Triton shells stained with uranyl acetate show the presence of numerous filaments which appear beaded and are 80--120 A in diameter. The filaments cannot be composed mainly af actin, but enough spectrin is present to form the filaments. Triton shells may provide an excellent source of material useful in the investigation of the erythrocyte cytoskeleton.

  6. Effect of honokiol on erythrocytes.

    PubMed

    Zbidah, Mohanad; Lupescu, Adrian; Herrmann, Tabea; Yang, Wenting; Foller, Michael; Jilani, Kashif; Lang, Florian

    2013-09-01

    Honokiol ((3,5-di-(2-propenyl)-1,1-biphenyl-2,2-diol), a component of Magnolia officinalis, stimulates apoptosis and is thus considered for the treatment of malignancy. In analogy to apoptosis of nucleated cells, erythrocytes may enter eryptosis, a suicidal death characterized by cell shrinkage and by breakdown of cell membrane phosphatidylserine asymmetry with phosphatidylserine-exposure at the erythrocyte surface. Eryptosis may be triggered following increase of cytosolic Ca(2+)-activity ([Ca(2+)]i). The present study explored, whether honokiol elicits eryptosis. Cell volume has been estimated from forward scatter, phosphatidylserine-exposure from annexin V binding, hemolysis from hemoglobin release, [Ca(2+)]i from Fluo3-fluorescence, and ceramide from fluorescent antibodies. As a result, a 48 h exposure to honokiol was followed by a slight but significant increase of [Ca(2+)]i (15 μM), significant decrease of forward scatter (5 μM), significant increase of annexin-V-binding (5 μM) and significant increase of ceramide formation (15 μM). Honokiol further induced slight, but significant hemolysis. Honokiol (15 μM) induced annexin-V-binding was significantly blunted but not abrogated in the nominal absence of extracellular Ca(2+). In conclusion, honokiol triggers suicidal erythrocyte death or eryptosis, an effect at least in part due to stimulation of Ca(2+) entry and ceramide formation. Copyright © 2013 Elsevier Ltd. All rights reserved.

  7. Evaluation of Intravascular Hemolysis With Erythrocyte Creatine in Patients With Aortic Stenosis.

    PubMed

    Sugiura, Tetsuro; Okumiya, Toshika; Kubo, Toru; Takeuchi, Hiroaki; Matsumura, Yoshihisa

    2016-07-27

    Chronic intravascular hemolysis has been identified in patients with cardiac valve prostheses, but only a few case reports have evaluated intravascular hemolysis in patients with native valvular heart disease. To detect intravascular hemolysis in patients with aortic stenosis, erythrocyte creatine was evaluated with hemodynamic indices obtained by echocardiography.Erythrocyte creatine, a marker of erythrocyte age, was assayed in 30 patients with aortic stenosis and 10 aged matched healthy volunteers. Peak flow velocity of the aortic valve was determined by continuous-wave Doppler echocardiography. Twenty of 30 patients with aortic stenosis had high erythrocyte creatine levels (> 1.8 µmol/g Hb) and erythrocyte creatine was significantly higher as compared with control subjects (1.98 ± 0.49 versus 1.52 ± 0.19 µmol/g Hb, P = 0.007). Peak transvalvular pressure gradient ranged from 46 to 142 mmHg and peak flow velocity ranged from 3.40 to 5.95 m/second. Patients with aortic stenosis had a significantly lower erythrocyte count (387 ± 40 versus 436 ± 42 × 10(4) µL, P = 0.002) and hemoglobin (119 ± 11 versus 135 ± 11 g/L, P < 0.001) as compared with control subjects. Erythrocyte creatine had a fair correlation with peak flow velocity (r = 0.55, P = 0.002).In conclusion, intravascular hemolysis due to destruction of erythrocytes was detected in patients with moderate to severe aortic stenosis and the severity of intravascular hemolysis was related to valvular flow velocity of the aortic valve.

  8. [Erythrocytic parameters Sysmex in a case of severe haemolysis].

    PubMed

    Ferrero-Vacher, Corinne; Senlis, Jean-Éric; Loustaunau, Denis; Aquaronne, Danièle; Jeandel, Pierre-Yves; Sudaka, Isabelle

    2015-01-01

    We are reporting a case of severe haemolytic anemia with cold agglutinins which combines several spurious determinations. It shows the usefulness of the new erythrocytic parameters of the XE 5000 Sysmex, specially: red blood cells with optical count (RBC-O), GR-He (intra-erythocytic hemoglobin) and R-MFV (most frequent volume). Optical red blood cells act as a substitute for red cells count instead of impedance red cells and R-MFV as a substitute for MCV (mean cell volume). The hematocrit (HCT) is corrected thanks to the following formula: HCT=(RBC-O X R- MFV)/1000. Free plasmatic hemoglobin is included in the measure of hemoglobin by the analyzer but is not available for tissue oxygenation. So, hemoglobin (HGB) has to be corrected by the means of GR- He thanks to the following formula: HGB=(GR He x RBC-O)/10.

  9. [AGGREGATION OF METABOLICALLY DEPLETED HUMAN ERYTHROCYTES].

    PubMed

    Sheremet'ev, Yu A; Popovicheva, A N; Rogozin, M M; Levin, G Ya

    2016-01-01

    An aggregation of erythrocytes in autologous plasma after blood storage for 14 days at 4 °C was studied using photometry and light microscopy. The decrease of ATP content, the formation of echinocytes and spheroechinocytes, the decrease of rouleaux form of erythrocyte aggregation were observed during the storage. On the other hand the aggregates of echinocytes were formed in the stored blood. The addition of plasma from the fresh blood didn't restore the normal discocytic shape and aggregation of erythrocytes in the stored blood. The possible mechanisms of erythrocytes and echinocytes aggregation are discussed.

  10. Nuclear magnetic resonance investigation of erythrocyte membranes in chronic myeloproliferative disorders.

    PubMed

    Morariu, V V; Petrov, L

    1986-07-01

    The temperature dependence of the apparent water diffusional exchange through erythrocyte membranes in cases of policitemia vera, chronic granulocytic leukemia and primary myelofibrosis was measured by using a nuclear magnetic resonance method in the presence of Mn2+. The thermal transition shifted to lower temperatures in all cases, regardless of the stage of the disease, suggesting a structural alteration of the membrane. The shift of transition indirectly suggests a lower penetration of the erythrocytes by Mn2+. The water exchange time at 37 degrees C also increased, mainly in the blast crisis; it seems to have a prognostic value of some clinical interest. No simple correlation of the water exchange and the following clinical investigations was observed: the white count, the percentage of promyelocites and myeloblasts, the sedimentation rate of blood, the osmotic fragility of erythrocytes, the total concentration of proteins, albumin and immunoglobulins, respectively, in plasma.

  11. A side-by-side evaluation of four platelet-counting instruments.

    PubMed

    Dalton, W T; Bollinger, P; Drewinko, B

    1980-08-01

    The performances of four instruments for counting platelets were evaluated in a side-by-side study: the Haema-Count MK-4/HC, an electronic impedance instrument that counts platelets in platelet-rich plasma; the Ultra-Flo 100, and the Coulter Counter Model S-Plus, electronic impedance instruments that count platelets in the presence of intact erythrocytes; and the AutoCounter, an optical instrument that counts platelets in the presence of lysed erythrocytes. The Ultra-Flo 100 and the S-Plus showed the best within-run precision, and all four instruments were considerably more precise than manual platelet counting, especially at low levels of platelet count. The four instruments were all linear in the ranges tested (5 to 650 x 10(9)/or greater), and sample carry-over was less than 0.7% for each. A noteworthy finding was that the erythrocyte concentration of the blood samples affected the displayed platelet count of the S-Plus and, to a lesser extent, that of the AutoCounter, in a predictable way, whereas it did not greatly affect the displayed count of the Ultra-Flo 100. In addition to differences in quality of performances, the four instruments differed considerably in speed and ease of operation and in cost.

  12. Effect of thioridazine on erythrocytes.

    PubMed

    Lang, Elisabeth; Modicano, Paola; Arnold, Markus; Bissinger, Rosi; Faggio, Caterina; Abed, Majed; Lang, Florian

    2013-10-23

    Thioridazine, a neuroleptic phenothiazine with antimicrobial efficacy is known to trigger anemia. At least in theory, the anemia could result from stimulation of suicidal erythrocyte death or eryptosis, which is characterized by cell shrinkage and by phospholipid scrambling of the cell membrane with phosphatidylserine exposure at the erythrocyte surface. Triggers of eryptosis include increase of cytosolic Ca²⁺-concentration ([Ca²⁺](i)) and activation of p38 kinase. The present study explored, whether thioridazine elicits eryptosis. [Ca²⁺](i) has been estimated from Fluo3-fluorescence, cell volume from forward scatter, phosphatidylserine exposure from annexin-V-binding, and hemolysis from hemoglobin release. A 48 hours exposure to thioridazine was followed by a significant increase of [Ca²⁺](i) (30 µM), decrease of forward scatter (30 µM), and increase of annexin-V-binding (≥12 µM). Nominal absence of extracellular Ca²⁺ and p38 kinase inhibitor SB203580 (2 µM) significantly blunted but did not abolish annexin-V-binding following thioridazine exposure. Thioridazine stimulates eryptosis, an effect in part due to entry of extracellular Ca²⁺ and activation of p38 kinase.

  13. Enzymatic Production of Universal Donor Erythrocytes.

    DTIC Science & Technology

    1981-10-01

    strong activities of extracellular glycosidases that convert blood type A or B erythrocytes to universal donor blood type O erythrocytes; 2) to purify the... blood type B-degrading enzyme produced by a fecal strain of Ruminococcus AB; 3) to determine whether human type B red cells could be safety converted

  14. Induction of Suicidal Erythrocyte Death by Cantharidin.

    PubMed

    Alzoubi, Kousi; Egler, Jasmin; Briglia, Marilena; Fazio, Antonella; Faggio, Caterina; Lang, Florian

    2015-07-28

    The natural phosphoprotein phosphatase inhibitor cantharidin, primarily used for topical treatment of warts, has later been shown to trigger tumor cell apoptosis and is thus considered for the treatment of malignancy. Similar to apoptosis of tumor cells, erythrocytes may undergo eryptosis, a suicidal cell death characterized by cell shrinkage and translocation of cell membrane phosphatidylserine to the erythrocyte surface. Signaling of eryptosis includes increase of cytosolic Ca2+-activity ([Ca2+]i), ceramide, oxidative stress and dysregulation of several kinases. Phosphatidylserine abundance at the erythrocyte surface was quantified utilizing annexin-V-binding, cell volume from forward scatter, [Ca2+]i from Fluo3-fluorescence, ceramide from antibody binding, and reactive oxidant species (ROS) from 2',7'-dichlorodihydrofluorescein diacetate (DCFDA) fluorescence. A 48 h treatment of human erythrocytes with cantharidin significantly increased the percentage of annexin-V-binding cells (≥10 mg/mL), significantly decreased forward scatter (≥25 mg/mL), significantly increased [Ca2+]i (≥25 mg/mL), but did not significantly modify ceramide abundance or ROS. The up-regulation of annexin-V-binding following cantharidin treatment was not significantly blunted by removal of extracellular Ca2+ but was abolished by kinase inhibitor staurosporine (1 mM) and slightly decreased by p38 inhibitor skepinone (2 mM). Exposure of erythrocytes to cantharidin triggers suicidal erythrocyte death with erythrocyte shrinkage and erythrocyte membrane scrambling, an effect sensitive to kinase inhibitors staurosporine and skepinone.

  15. Induction of suicidal erythrocyte death by nelfinavir.

    PubMed

    Bissinger, Rosi; Waibel, Sabrina; Lang, Florian

    2015-05-08

    The HIV protease inhibitor, nelfinavir, primarily used for the treatment of HIV infections, has later been shown to be effective in various infectious diseases including malaria. Nelfinavir may trigger mitochondria-independent cell death. Erythrocytes may undergo eryptosis, a mitochondria-independent suicidal cell death characterized by cell shrinkage and phosphatidylserine translocation to the erythrocyte surface. Triggers of eryptosis include oxidative stress and increase of cytosolic Ca2+-activity ([Ca2+]i). During malaria, accelerated death of infected erythrocytes may decrease parasitemia and thus favorably influence the clinical course of the disease. In the present study, phosphatidylserine abundance at the cell surface was estimated from annexin V binding, cell volume from forward scatter, reactive oxidant species (ROS) from 2',7'-dichlorodihydrofluorescein diacetate (DCFDA) fluorescence, and [Ca2+]i from Fluo3-fluorescence. A 48 h treatment of human erythrocytes with nelfinavir significantly increased the percentage of annexin-V-binding cells (≥5µg/mL), significantly decreased forward scatter (≥2.5µg/mL), significantly increased ROS abundance (10 µg/mL), and significantly increased [Ca2+]i (≥5 µg/mL). The up-regulation of annexin-V-binding following nelfinavir treatment was significantly blunted, but not abolished by either addition of the antioxidant N-acetylcysteine (1 mM) or removal of extracellular Ca2+. In conclusion, exposure of erythrocytes to nelfinavir induces oxidative stress and Ca2+ entry, thus leading to suicidal erythrocyte death characterized by erythrocyte shrinkage and erythrocyte membrane scrambling.

  16. All about Carbohydrate Counting

    MedlinePlus

    Toolkit No. 14 All About Carbohydrate Counting What is carbohydrate counting? Carbohydrate counting is a way to plan your meals. It can help ... Diabetes Association, Inc. 2/14 Toolkit No. 14: All About Carbohydrate Counting continued The chart at the ...

  17. Assessment of radio-frequency electromagnetic radiation by the micronucleus test in bovine peripheral erythrocytes.

    PubMed

    Balode, Z

    1996-02-02

    Previous bioindicative studies in the Skrunda Radio Location Station area have focused on the somatic influence of electromagnetic radiation on plants, but it is also important to study genetic effects. We have chosen cows as test animals for cytogenetical evaluation because they live in the same general exposure area as humans, are confined to specific locations and are chronically exposed to radiation. Blood samples were obtained from female Latvian Brown cows from a farm close to and in front of the Skrunda Radar and from cows in a control area. A simplified alternative to the Schiff method of DNA staining for identification of micronuclei in peripheral erythrocytes was applied. Microscopically, micronuclei in peripheral blood erythrocytes were round in shape and exhibited a strong red colour. They are easily detectable as the only coloured bodies in the uncoloured erythrocytes. From each individual animal 2000 erythrocytes were examined at a magnification of x 1000 for the presence of micronuclei. The counting of micronuclei in peripheral erythrocytes gave low average incidences, 0.6 per 1000 in the exposed group and 0.1 per 1000 in the control, but statistically significant (P < 0.01) differences were found in the frequency distribution between the control and exposed groups.

  18. Changes in haematology, plasma biochemistry and erythrocyte osmotic fragility of the Nigerian laughing dove (Streptopelia senegalensis) in captivity.

    PubMed

    Azeez, O I; Oyagbemi, A A; Olawuwo, O S; Oyewale, J O

    2013-06-30

    The haematology, plasma biochemistry and erythrocyte osmotic fragility of the Nigerian laughing dove (Streptopelia senegalensis) were studied after 4 and 8 weeks in captivity. At 8 weeks, there was a normocytic hypochromic anaemia characterized by reduced values for packed cell volume (PCV), red blood cell count (RBC), haemoglobin (Hb) concentration, mean corpuscular haemoglobin (MCH) and mean corpuscular haemoglobin concentration (MCHC), but the mean corpuscular volume (MCV) was unaltered compared with the corresponding values at 4 weeks. The platelet count, total white blood cell count, heterophil, lymphocyte and monocyte counts were also lower at 8 weeks than those of the birds sampled at 4 weeks in captivity. There was also a stress induced increased heterophil/lymphocyte ratio and the erythrocytes were more fragile in hypotonic solution in birds sampled at 8 weeks. Plasma aspartate transaminase (AST), alanine aminotransferase (ALT) and alkaline phosphate (ALP) increased at 8 weeks, though non-significantly, which might have been due to muscle wasting consequent upon decreased muscular activities associated with prolonged captivity. The results suggest that maintaining wild birds in captivity for a prolonged period could be stressful as shown by the heterophil/lymphocytes ratio and reduced erythrocyte osmotic resistance, and could lead to decreases in erythrocyte parameters and muscle wasting.

  19. Accuracy of a New Platelet Count System (PLT-F) Depends on the Staining Property of Its Reagents

    PubMed Central

    Wada, Atsushi; Takagi, Yuri; Kono, Mari; Morikawa, Takashi

    2015-01-01

    Background Platelet count is essential for the diagnosis and management of hemostasis abnormalities. Although existing platelet count methods installed in common hematology analyzers can correctly count platelets in normal blood samples, they tend to miscount platelets in some abnormal samples. The newly developed PLT-F channel in the XN-Series hematology analyzer (Sysmex) has been reported to be a reliable platelet count system, even in abnormal samples. However, how the PLT-F platelet counting system achieves such accuracy has not been described in scientific articles. Methods Isolated platelets, erythrocytes, and fragmented erythrocytes were examined using an automated hematology analyzer. The samples were labeled by combining PLT-F reagents and anti-CD62p, CD63, Grp75, Calreticulin, CD41, or CD61 antibody, and analyzed using confocal laser scanning microscopy or flow cytometry. Results The PLT-F system correctly discriminated platelets in erythrocytes. Its reagents strongly stained some intraplatelet organelles labeled with anti-Grp75, but only faintly stained the plasma membrane of both platelets and erythrocytes. Microscopic observation and flow cytometric examination revealed that all of these strongly stained cells were also labeled with platelet-specific anti-CD41 and anti-CD61 antibodies. Conclusions This study revealed that the staining property of the PLT-F reagents, by which platelets and fragmented erythrocytes are clearly distinguished, contributes to the platelet-counting accuracy of the PLT-F system. PMID:26496387

  20. Molecular basis for erythrocyte shape

    NASA Astrophysics Data System (ADS)

    Elgsaeter, A.; Mikkelsen, A.

    1991-05-01

    The isolated plasma membrane of the human erythrocytes displays the same shape and shape transformations as the intact cells. It is therefore generally believed that the plasma membrane plays a dominant role in determining erythrocyte shape. The plasma membrane consists of a fluid lipid bilayer to the surface of which is attached a protein skeleton. The two halves of the lipid bilayer and the protein network (gel) are tighly coupled, but at the same time elastically deformable and can slide relative to one another in the plane of the cell membrane. The equilibrium shape of such a structure is determined by the combined mechano-chemical properties of the individual layers and equals the cell shape that for the given cell volume corresponds to the lowest total elastic free energy. The elastic free energy of the lipid bilayer is mainly associated with bending and change in surface area for each of the two lipid monolayer. For the protein membrane skeleton the elastic free energy mainly equals the sum of the local contributions due to shear deformation and surface change. When the mechano-chemical properties of each of the layers are known, calculation of the equilibrium shape is in principle just an exercise in standard continuum mechanics. The elastic properties of pure lipid monolayers have long been qualitatively fairly well known. The changes in lipid bilayer elastic properties resulting from the presence of integral membrane proteins have just recently become better understood. The detailed molecular basis for the elastic properties of the protein membrane skeleton remains unresolved despite many attempts to elucidate the problem. It is widely agreed that the elastic properties are largely accounted for by the highly elongated spectrin molecules, but whether the membrane skelton elasticity is mainly of entropic or entalphic origin is still unsettled.

  1. [Lysophosphatidic acid and human erythrocyte aggregation].

    PubMed

    Sheremet'ev, Iu A; Popovicheva, A N; Levin, G Ia

    2014-01-01

    The effects of lysophosphatidic acid on the morphology and aggregation of human erythrocytes has been studied. Morphology of erythrocytes and their aggregates were studied by light microscopy. It has been shown that lysophosphatidic acid changes the shape of red blood cells: diskocyte become echinocytes. Aggregation of red blood cells (rouleaux) was significantly reduced in autoplasma. At the same time there is a strong aggregation of echinocytes. This was accompanied by the formation of microvesicles. Adding normal plasma to echinocytes restores shape and aggregation of red blood cells consisting of "rouleaux". A possible mechanism of action of lysophosphatidic acid on erythrocytes is discussed.

  2. Gravity sedimentation of leukocytes is partially independent from erythrocyte sedimentation.

    PubMed

    Bogar, L L; Tarsoly, P P

    2006-01-01

    Leukocyte function tests are suitable for monitoring the severity of chronic inflammatory and acute infectious diseases. The tests usually require time consuming leukocyte separation techniques while the original character of leukocytes can substantially alter. In contrast, we noted that gravity sedimentation properties of leukocytes is simple to measure and it also reflects non-specific inflammatory reactions of leukocytes. Our novel test is named leukocyte antisedimentation rate (LAR) which is measured by leukocyte counting in the upper (U) and lower (L) half of the sedimentation blood column after one-hour gravity sedimentation of the whole blood. The formula LAR=100.(U-L)/(U+L) is used to calculate the percentage of leukocytes crosses the middle line of sedimentation blood column upward during one-hour sedimentation (normal range<15%, inter-assay coefficient of variation<5%). In this study we found that in vitro pre-treatment of septic patients' blood samples with protamine, lidocaine and prednisolone decreased leukocyte antisedimentation rate in a concentration dependent manner without effecting erythrocyte sedimentation rate. Leukocyte adherence was measured by the retention rate of leukocytes in a nylon fibre column. There was a significant positive correlation between leukocyte antisedimentation rate and leukocyte adherence (p<0.01), hematocrit (p<0.05), erythrocyte sedimentation rate (p<0.05) when blood samples of 35 healthy individuals were analysed. We concluded that leukocyte antisedimentation rate in septic patients is significantly elevated comparing to healthy controls and as a bedside test it can reflect leukocyte involvement in infections. In vitro protamine, lidocaine and prednisolone pre-treatment of septic patients' blood samples indicates that leukocyte antisedimentation process is partially independent from the ongoing erythrocyte sedimentation.

  3. The Big Pumpkin Count.

    ERIC Educational Resources Information Center

    Coplestone-Loomis, Lenny

    1981-01-01

    Pumpkin seeds are counted after students convert pumpkins to jack-o-lanterns. Among the activities involved, pupils learn to count by 10s, make estimates, and to construct a visual representation of 1,000. (MP)

  4. The Big Pumpkin Count.

    ERIC Educational Resources Information Center

    Coplestone-Loomis, Lenny

    1981-01-01

    Pumpkin seeds are counted after students convert pumpkins to jack-o-lanterns. Among the activities involved, pupils learn to count by 10s, make estimates, and to construct a visual representation of 1,000. (MP)

  5. The erythrocyte ghost is a perfect osmometer.

    PubMed

    Kwant, W O; Seeman, P

    1970-02-01

    The osmotic swelling of intact erythrocytes in hypotonic solutions was measured using microhematocrit tubes, Van Allen tubes, and a calibrated Coulter counter. In agreement with earlier workers the intact cells did not behave as perfect osmometers, the cells swelling less than predicted by the Boyle-van't Hoff law. Erythrocyte ghosts were prepared from fresh intact erythrocytes by one-step hemolysis in 0.25% NaCl at an extremely dilute concentration of cells and the membranes were sealed at 37 degrees . The ghosts were mixed with NaCl solutions of different osmolarities and the MCV (mean cell volume) of the shrunken cells immediately monitored by a calibrated Coulter counter. It was found that the MCV values of the shrunken ghosts were accurately predicted by the Boyle-van't Hoff law. These results indicate that these erythrocyte ghosts behaved as perfect osmometers.

  6. Characterization of lipid domains in erythrocyte membranes.

    PubMed

    Rodgers, W; Glaser, M

    1991-02-15

    Fluorescence digital imaging microscopy was used to study the lateral distribution of the lipid components in erythrocyte membranes. Intact erythrocytes labeled with phospholipids containing a fluorophore attached to one fatty acid chain showed an uneven distribution of the phospholipids in the membrane thereby demonstrating the presence of membrane domains. The enrichment of the lipotropic compound chlor-promazine in domains in intact erythrocytes also suggested that the domains are lipid-enriched regions. Similar membrane domains were present in erythrocyte ghosts. The phospholipid enrichment was increased in the domains by inducing membrane protein aggregation. Double-labeling experiments were done to determine the relative distributions of different phospholipids in the membrane. Vesicles made from extracted lipids did not show the presence of domains consistent with the conclusion that membrane proteins were responsible for creating the domains. Overall, it was found that large domains exist in the red blood cell membrane with unequal enrichment of the different phospholipid species.

  7. Induction of Suicidal Erythrocyte Death by Nelfinavir

    PubMed Central

    Bissinger, Rosi; Waibel, Sabrina; Lang, Florian

    2015-01-01

    The HIV protease inhibitor, nelfinavir, primarily used for the treatment of HIV infections, has later been shown to be effective in various infectious diseases including malaria. Nelfinavir may trigger mitochondria-independent cell death. Erythrocytes may undergo eryptosis, a mitochondria-independent suicidal cell death characterized by cell shrinkage and phosphatidylserine translocation to the erythrocyte surface. Triggers of eryptosis include oxidative stress and increase of cytosolic Ca2+-activity ([Ca2+]i). During malaria, accelerated death of infected erythrocytes may decrease parasitemia and thus favorably influence the clinical course of the disease. In the present study, phosphatidylserine abundance at the cell surface was estimated from annexin V binding, cell volume from forward scatter, reactive oxidant species (ROS) from 2',7'-dichlorodihydrofluorescein diacetate (DCFDA) fluorescence, and [Ca2+]i from Fluo3-fluorescence. A 48 h treatment of human erythrocytes with nelfinavir significantly increased the percentage of annexin-V-binding cells (≥5µg/mL), significantly decreased forward scatter (≥2.5µg/mL), significantly increased ROS abundance (10 µg/mL), and significantly increased [Ca2+]i (≥5 µg/mL). The up-regulation of annexin-V-binding following nelfinavir treatment was significantly blunted, but not abolished by either addition of the antioxidant N-acetylcysteine (1 mM) or removal of extracellular Ca2+. In conclusion, exposure of erythrocytes to nelfinavir induces oxidative stress and Ca2+ entry, thus leading to suicidal erythrocyte death characterized by erythrocyte shrinkage and erythrocyte membrane scrambling. PMID:26008229

  8. Alkali ion transport of primycin modified erythrocytes.

    PubMed

    Blaskó, K; Györgyi, S

    1981-01-01

    The effects of the antibiotic primycin on alkali cation transport of human erythrocytes were investigated. Primycin selectively increases the permeability of erythrocytes to alkali-cations according to the sequence: Cs+ greater than Rb+ approximately K+ greater than Na+. The time course of the cation effluxes depends on the antibiotic concentration and can be altered by negatively charged SDS. Some evidence is given for the mechanism of primycin-membrane interaction.

  9. Laser interference microscopy in erythrocyte study

    NASA Astrophysics Data System (ADS)

    Yusipovich, A. I.; Parshina, E. Yu.; Brysgalova, N. Yu.; Brazhe, A. R.; Brazhe, N. A.; Lomakin, A. G.; Levin, G. G.; Maksimov, G. V.

    2009-05-01

    With the laser interference microscopy (LIM) technique, one can measure phase height of cells—a variable proportional to the cell thickness and the difference in the refractive indices of the cell and the surrounding medium. This makes functional optical cell imaging possible, and estimation of shape, thickness, and area of erythrocytes feasible. In this paper, we studied changes in erythrocyte shape and volume with osmolarity and pH. Obtained from the LIM technique, erythrocyte phase heights and area values, as well as the hematocrit-measured erythrocyte volume, were used to estimate changes in the refractive index with osmolarity and pH. A comparison between the estimated refractive index with the refractive index, calculated in the assumption that it can only depend on the hemoglobin concentration in the cell, indicates that these two estimates are identical in the range of osmolarity (250-1000 mOsm) and pH (4.5-10.0) values. Thus, refractive index changes result exclusively from the changes in hemoglobin concentration with the changes in erythrocyte volume. Under these conditions, it is possible to estimate the amount of hemoglobin in an erythrocyte from its phase height and area, obtained from LIM.

  10. The Effect of Sepsis on the Erythrocyte.

    PubMed

    Bateman, Ryon M; Sharpe, Michael D; Singer, Mervyn; Ellis, Christopher G

    2017-09-08

    Sepsis induces a wide range of effects on the red blood cell (RBC). Some of the effects including altered metabolism and decreased 2,3-bisphosphoglycerate are preventable with appropriate treatment, whereas others, including decreased erythrocyte deformability and redistribution of membrane phospholipids, appear to be permanent, and factors in RBC clearance. Here, we review the effects of sepsis on the erythrocyte, including changes in RBC volume, metabolism and hemoglobin's affinity for oxygen, morphology, RBC deformability (an early indicator of sepsis), antioxidant status, intracellular Ca(2+) homeostasis, membrane proteins, membrane phospholipid redistribution, clearance and RBC O₂-dependent adenosine triphosphate efflux (an RBC hypoxia signaling mechanism involved in microvascular autoregulation). We also consider the causes of these effects by host mediated oxidant stress and bacterial virulence factors. Additionally, we consider the altered erythrocyte microenvironment due to sepsis induced microvascular dysregulation and speculate on the possible effects of RBC autoxidation. In future, a better understanding of the mechanisms involved in sepsis induced erythrocyte pathophysiology and clearance may guide improved sepsis treatments. Evidence that small molecule antioxidants protect the erythrocyte from loss of deformability, and more importantly improve septic patient outcome suggest further research in this area is warranted. While not generally considered a critical factor in sepsis, erythrocytes (and especially a smaller subpopulation) appear to be highly susceptible to sepsis induced injury, provide an early warning signal of sepsis and are a factor in the microvascular dysfunction that has been associated with organ dysfunction.

  11. Mechanisms of Human Erythrocytic Bioactivation of Nitrite*

    PubMed Central

    Liu, Chen; Wajih, Nadeem; Liu, Xiaohua; Basu, Swati; Janes, John; Marvel, Madison; Keggi, Christian; Helms, Christine C.; Lee, Amber N.; Belanger, Andrea M.; Diz, Debra I.; Laurienti, Paul J.; Caudell, David L.; Wang, Jun; Gladwin, Mark T.; Kim-Shapiro, Daniel B.

    2015-01-01

    Nitrite signaling likely occurs through its reduction to nitric oxide (NO). Several reports support a role of erythrocytes and hemoglobin in nitrite reduction, but this remains controversial, and alternative reductive pathways have been proposed. In this work we determined whether the primary human erythrocytic nitrite reductase is hemoglobin as opposed to other erythrocytic proteins that have been suggested to be the major source of nitrite reduction. We employed several different assays to determine NO production from nitrite in erythrocytes including electron paramagnetic resonance detection of nitrosyl hemoglobin, chemiluminescent detection of NO, and inhibition of platelet activation and aggregation. Our studies show that NO is formed by red blood cells and inhibits platelet activation. Nitric oxide formation and signaling can be recapitulated with isolated deoxyhemoglobin. Importantly, there is limited NO production from erythrocytic xanthine oxidoreductase and nitric-oxide synthase. Under certain conditions we find dorzolamide (an inhibitor of carbonic anhydrase) results in diminished nitrite bioactivation, but the role of carbonic anhydrase is abrogated when physiological concentrations of CO2 are present. Importantly, carbon monoxide, which inhibits hemoglobin function as a nitrite reductase, abolishes nitrite bioactivation. Overall our data suggest that deoxyhemoglobin is the primary erythrocytic nitrite reductase operating under physiological conditions and accounts for nitrite-mediated NO signaling in blood. PMID:25471374

  12. [Kinetics of Cu crossing human erythrocyte membrane].

    PubMed

    Dun, Zhu Ci Ren

    2014-12-01

    This study was aimed to investigate various factors influencing the proceduction of Cu(II) crossing human erythrocyte membrane, including concentration of Cu²⁺, pH value of the medium, temperature and time of incubation, and to derive kinetic equation of Cu(II) crossing human erythrocyte membrane. Suspension red blood cells were incubated by Cu²⁺, then content of Cu²⁺ crossed human erythrocyte membrane was determined by atomic absorption spectrometry under various conditions after digestion. The results showed that content of Cu²⁺ crossed human erythrocyte membrane increased with the increase of extracellular Cu²⁺ and enhancement of incubation temperature, and the content of Cu²⁺ crossed human erythrocyte membrane showed a increasing tendency when pH reached to 6.2-7.4, and to maximum at pH 7.4, then gradually decreased at range of pH 7.4-9.2. It is concluded that the Cu²⁺ crossing human erythrocyte has been confirmed to be the first order kinetics characteristics within 120 min, and the linear equation is 10³ × Y = 0.0497t +6.5992.

  13. Therapeutic potential of manipulating suicidal erythrocyte death.

    PubMed

    Lang, Florian; Jilani, Kashif; Lang, Elisabeth

    2015-01-01

    Eryptosis, the suicidal erythrocyte death, is characterized by erythrocyte shrinkage and phosphatidylserine translocation to the erythrocyte surface. Eryptosis is triggered by cell stress such as energy depletion and oxidative stress, by Ca(2+)-entry, ceramide, caspases, calpain and/or altered activity of several kinases. Phosphatidylserine-exposing erythrocytes adhere to the vascular wall and may thus impede microcirculation. Eryptotic cells are further engulfed by phagocytes and thus rapidly cleared from circulation. Stimulation of eryptosis contributes to anemia of several clinical conditions such as metabolic syndrome, diabetes, malignancy, hepatic failure, heart failure, uremia, hemolytic uremic syndrome, sepsis, fever, dehydration, mycoplasma infection, malaria, iron deficiency, sickle cell anemia, thalassemia, glucose-6-phosphate dehydrogenase deficiency and Wilson's disease. On the other hand, eryptosis with subsequent clearance of infected erythrocytes in malaria may counteract parasitemia. In theory, anemia due to excessive eryptosis could be alleviated by treatment with small molecules inhibiting eryptosis. In malaria, stimulators of eryptosis may accelerate death of infected erythrocytes and thus favorably influence the clinical course of the disease. Many small molecules inhibit or stimulate eryptosis. Several stimulators favorably influence murine malaria. Further preclinical and subsequent clinical studies are required to elucidate the therapeutic potential of stimulators or inhibitors of eryptosis.

  14. Bile Acid-Induced Suicidal Erythrocyte Death.

    PubMed

    Lang, Elisabeth; Pozdeev, Vitaly I; Gatidis, Sergios; Qadri, Syed M; Häussinger, Dieter; Kubitz, Ralf; Herebian, Diran; Mayatepek, Ertan; Lang, Florian; Lang, Karl S; Lang, Philipp A

    2016-01-01

    In nucleated cells, bile acids may activate cation channels subsequently leading to entry of Ca2+. In erythrocytes, increase of cytosolic Ca2+ activity triggers eryptosis, the suicidal death of erythrocytes characterized by phosphatidylserine exposure at the cell surface and cell shrinkage. Eryptosis is triggered by bile duct ligation, an effect partially attributed to conjugated bilirubin. The present study explored, whether bile acids may stimulate eryptosis. Phosphatidylserine exposing erythrocytes have been identified utilizing annexin V binding, cell volume estimated from forward scatter, cytosolic Ca2+ activity determined using Fluo-3 fluorescence, and ceramide abundance at the erythrocyte surface utilizing specific antibodies. The exposure of human erythrocytes to glycochenodesoxycholic (GCDC) and taurochenodesoxycholic (TCDC) acid was followed by a significant decrease of forward scatter and significant increase of Fluo-3 fluorescence, ceramide abundance as well as annexin V binding. The effect on annexin V binding was significantly blunted, but not abolished by removal of extracellular Ca2+. Bile acids stimulate suicidal cell death, an effect paralleled by and in part due to Ca2+ entry and ceramide. The bile acid induced eryptosis may in turn lead to accelerated clearance of circulating erythrocytes and, thus, may contribute to anemia in cholestatic patients. © 2016 The Author(s) Published by S. Karger AG, Basel.

  15. Adenosine deaminase activity in serum, erythrocytes and lymphocytes of rats infected with Leptospira icterohaemorrhagiae.

    PubMed

    Tonin, Alexandre A; Pimentel, Victor C; da Silva, Aleksandro S; de Azevedo, Maria Isabel; Souza, Viviane C G; Wolkmer, Patrícia; Rezer, João F P; Badke, Manoel R T; Leal, Daniela B R; Schetinger, Maria Rosa C; Monteiro, Silvia G; Lopes, Sonia T A

    2012-04-01

    Leptospirosis is a systemic disease of humans and domestic animals, mainly dogs, cattle and swine. The course of human leptospirosis varies from mild to severe fatal forms and the most severe form of human leptospirosis is principally caused by Leptospira interrogans serovar icterohaemorrhagiae (L. icterohaemorrhagiae). The enzyme adenosine deaminase (ADA) plays an important role in the production and differentiation of blood cells. The aim of this study was to evaluate the activity of ADA in serum, erythrocytes and lymphocytes of rats infected with L. icterohaemorrhagiae, as compared with non-infected rats. Twenty-four adult rats, divided into two uniform groups (A and B) were used for the enzymatic assays. The animals in Group B were inoculated intraperitoneally with 2×10(8) leptospires/rat, and the rodents in Group A (control) were not-inoculated. Blood collection was performed on days 5 and 15 post-infection (PI) and the blood used to assess the ADA activity. The infection by L.icterohaemorrhagiae altered erythrocyte count, hemoglobin concentration and hematocrit, causing a decrease in all these parameters on day 15 PI. Lymphocytes decreased significantly on day 15 PI, and ADA activity in serum was inhibited in infected rats on days 5 and 15 PI and its activity in erythrocytes were increased on day 5 PI. On day 5 PI, we found an increase in ADA activity in erythrocytes of infected rats. No correlation was observed between hematocrit and erythrocyte ADA activity on days 5 and 15 PI. The ADA activity was inhibited in rats infected on day 15 PI. A positive correlation (r(2)=60) was also observed between the number of lymphocytes and ADA activity in lymphocytes on day 15 PI (P<0.05). In conclusion, our results showed that the ADA activity is altered in serum, lymphocytes and erythrocytes in experimental infection by L.icterohaemorrhagiae in rats, concomitantly with hematological parameters.

  16. Platelet and not erythrocyte microparticles are procoagulant in transfused thalassaemia major patients.

    PubMed

    Agouti, Imane; Cointe, Sylvie; Robert, Stéphane; Judicone, Coralie; Loundou, Anderson; Driss, Fathi; Brisson, Alain; Steschenko, Dominique; Rose, Christian; Pondarré, Corinne; Bernit, Emmanuelle; Badens, Catherine; Dignat-George, Françoise; Lacroix, Romaric; Thuret, Isabelle

    2015-11-01

    The level of circulating platelet-, erythrocyte-, leucocyte- and endothelial-derived microparticles detected by high-sensitivity flow cytometry was investigated in 37 β-thalassaemia major patients receiving a regular transfusion regimen. The phospholipid procoagulant potential of the circulating microparticles and the microparticle-dependent tissue factor activity were evaluated. A high level of circulating erythrocyte- and platelet-microparticles was found. In contrast, the number of endothelial microparticles was within the normal range. Platelet microparticles were significantly higher in splenectomized than in non-splenectomized patients, independent of platelet count (P < 0·001). Multivariate analysis indicated that phospholipid-dependent procoagulant activity was influenced by both splenectomy (P = 0·001) and platelet microparticle level (P < 0·001). Erythrocyte microparticles were not related to splenectomy, appear to be devoid of proper procoagulant activity and no relationship between their production and haemolysis, dyserythropoiesis or oxidative stress markers could be established. Intra-microparticle labelling with anti-HbF antibodies showed that they originate only partially (median of 28%) from thalassaemic erythropoiesis. In conclusion, when β-thalassaemia major patients are intensively transfused, the procoagulant activity associated with thalassaemic erythrocyte microparticles is probably diluted by transfusions. In contrast, platelet microparticles, being both more elevated and more procoagulant, especially after splenectomy, may contribute to the residual thrombotic risk reported in splenectomized multi-transfused β-thalassaemia major patients. © 2015 John Wiley & Sons Ltd.

  17. Characterization of carrier erythrocytes for biosensing applications.

    PubMed

    Bustamante López, Sandra C; Meissner, Kenith E

    2017-09-01

    Erythrocyte abundance, mobility, and carrying capacity make them attractive as a platform for blood analyte sensing as well as for drug delivery. Sensor-loaded erythrocytes, dubbed erythrosensors, could be reinfused into the bloodstream, excited noninvasively through the skin, and used to provide measurement of analyte levels in the bloodstream. Several techniques to load erythrocytes, thus creating carrier erythrocytes, exist. However, their cellular characteristics remain largely unstudied. Changes in cellular characteristics lead to removal from the bloodstream. We hypothesize that erythrosensors need to maintain native erythrocytes’ (NEs) characteristics to serve as a long-term sensing platform. Here, we investigate two loading techniques and the properties of the resulting erythrosensors. For loading, hypotonic dilution requires a hypotonic solution while electroporation relies on electrical pulses to perforate the erythrocyte membrane. We analyze the resulting erythrosensor signal, size, morphology, and hemoglobin content. Although the resulting erythrosensors exhibit morphological changes, their size was comparable with NEs. The hypotonic dilution technique was found to load erythrosensors much more efficiently than electroporation, and the sensors were loaded throughout the volume of the erythrosensors. Finally, both techniques resulted in significant loss of hemoglobin. This study points to the need for continued development of loading techniques that better preserve NE characteristics.

  18. Drug-induced erythrocyte membrane internalization.

    PubMed

    Ben-Bassat, I; Bensch, K G; Schrier, S L

    1972-07-01

    In vitro erythrocyte membrane internalization, resulting in the formation of membrane-lined vacuoles, can be quantified by a radioisotopic method. A complex of (37)Co-labeled vitamin B(12) and its plasma protein binders is first adsorbed to the cell surface, and after vacuoles are formed, the noninternalized label is removed by washing and trypsin treatment. The residual radioactivity represents trapped label and can be used to measure the extent of membrane internalization. Using this method, it was found that in addition to primaquine, a group of membrane-active drugs, specifically hydrocortisone, vinblastine, and chlorpromazine can induce membrane internalization in erythrocytes. This is a metabolic process dependent on drug concentration, temperature, and pH. Vacuole formation by all agents tested can be blocked by prior depletion of endogenous substrates or by poisoning the erythrocytes with sodium fluoride and sulfhydryl blocking agents. This phenomenon resembles in some respects the previously reported membrane internalization of energized erythrocyte ghosts. It is suggested that membrane internalization is dependent on an ATP-energized state and is influenced by the balance between the concentrations of magnesium and calcium in the membrane. This study provides a basis for proposing a unifying concept of the action of some membrane-active drugs, and for considering the role of erythrocyte membrane internalization in pathophysiologic events.

  19. Diffusion of glycophorin A in human erythrocytes.

    PubMed

    Giger, Katie; Habib, Ibrahim; Ritchie, Ken; Low, Philip S

    2016-11-01

    Several lines of evidence suggest that glycophorin A (GPA) interacts with band 3 in human erythrocyte membranes including: i) the existence of an epitope shared between band 3 and GPA in the Wright b blood group antigen, ii) the fact that antibodies to GPA inhibit the diffusion of band 3, iii) the observation that expression of GPA facilitates trafficking of band 3 from the endoplasmic reticulum to the plasma membrane, and iv) the observation that GPA is diminished in band 3 null erythrocytes. Surprisingly, there is also evidence that GPA does not interact with band 3, including data showing that: i) band 3 diffusion increases upon erythrocyte deoxygenation whereas GPA diffusion does not, ii) band 3 diffusion is greatly restricted in erythrocytes containing the Southeast Asian Ovalocytosis mutation whereas GPA diffusion is not, and iii) most anti-GPA or anti-band 3 antibodies do not co-immunoprecipitate both proteins. To try to resolve these apparently conflicting observations, we have selectively labeled band 3 and GPA with fluorescent quantum dots in intact erythrocytes and followed their diffusion by single particle tracking. We report here that band 3 and GPA display somewhat similar macroscopic and microscopic diffusion coefficients in unmodified cells, however perturbations of band 3 diffusion do not cause perturbations of GPA diffusion. Taken together the collective data to date suggest that while weak interactions between GPA and band 3 undoubtedly exist, GPA and band 3 must have separate interactions in the membrane that control their lateral mobility.

  20. Complexation of arsenic species in rabbit erythrocytes.

    PubMed

    Delnomdedieu, M; Basti, M M; Styblo, M; Otvos, J D; Thomas, D J

    1994-01-01

    The binding of arsenite, As(III), and arsenate, As(V), by molecules in the intracellular compartment of rabbit erythrocytes has been studied by 1H- and 31P-NMR spectroscopy, uptake of 73As, and ultrafiltration experiments. For intact erythrocytes to which 0.1-0.4 mM arsenite was added, direct evidence was obtained for entry of 76% within 1/2 h and subsequent binding of As(III) by intracellular glutathione and induced changes in the hemoglobin structure (NMR), likely due to binding of As(III). These results were compared with the effect of addition of As(V) on intact erythrocytes and revealed that a smaller amount of As(V) (approximately 25%) enters the cells; the main fraction of As(V) enters the phosphate pathway, depletes ATP, and increases Pi. In contrast, As(III) did not affect the ATP level. Both 1H- and 31P-NMR data indicated striking differences between As(III) and As(V) behavior when incubated with rabbit erythrocytes. These differences were confirmed by 73As uptake and binding experiments. meso-2,3-Dimercaptosuccinic acid (DMSA), a dithiol ligand, released glutathione from its arsenite complexes in erythrocytes.

  1. [Erythrocytes - the new application in medicine].

    PubMed

    Szumiło, Michał

    2013-01-01

    The new forms of drugs with better proprieties from traditional ones were sought for a long time. Erythrocytes applied as carriers of therapeutic substances are among promising. They are characterized by slower release of active substances, less toxicity, as well as better biocompatibility and biodegradation in the organism. It is especially important in administration of drugs with numerous side effects in therapy of chronic diseases e.g. malignancies. Investigations conducted from over twenty years showed, that erythrocytes are universal carriers in which different therapeutic substances were successfully closed, e.g. cytostatics, antibiotics, hormones and vitamins, as well as enzymes and vaccines. Some of the erythrocyte drug delivery systems are now studied at the clinical level, e.g. dexamthasone 21-phosphate in treatment of inflammatory bowel disease and chronic obstructive pulmonary disease. This substance encapsulated in human erythrocytes was also officially registered by European Medicines Agency, as the orphan drug in treatment of cystic fibrosis: Reports on application of carrier erythrocytes in patients with rare genetic diseases have also appeared.

  2. D-amino acids in aging erythrocytes.

    PubMed

    Ingrosso, D; Perna, A F

    1998-01-01

    Mature human erythrocytes are highly differentiated cells which have lost the ability to biosynthesize proteins de novo. During cell aging in circulation, erythrocyte proteins undergo spontaneous postbiosynthetic modifications, regarded as "protein fatigue" damage, which include formation of isomerized and/or racemized aspartyl residues. These damaged proteins cannot be replaced by new molecules; nevertheless, data support the notion that they can be repaired to a significant extent, through an enzymatic transmethylation reaction. This repair reaction has therefore been used as a means to monitor the increase of altered aspartyl residues in erythrocyte membrane proteins during cell aging. The relationship between protein repair and aspartyl racemization in red blood cell stress and disease is discussed.

  3. Effect of lead on erythrocyte membranes.

    PubMed Central

    Fukumoto, K; Karai, I; Horiguchi, S

    1983-01-01

    The effect of blood lead on erythrocyte membrane proteins was studied in 28 workers from a scrap lead refining factory and in 18 controls working in railway construction. Sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) of the polypeptides in the erythrocyte membrane showed that bands 3 and 4.1 had significantly decreased while bands 2.3, 6, and 7 had significantly increased in the lead workers compared with the controls. For the lead workers, the correlation coefficients between blood lead and bands 2.3 and 3 were r = 0.545 (p less than 0.01) and r = -0.51 (p less than 0.01) respectively. These results suggest that the decrease in erythrocyte membrane permeability results from a decrease in the membrane transfer protein responsible for band 3. Images PMID:6830722

  4. Mapping of hemoglobin in erythrocytes and erythrocyte ghosts using two photon excitation fluorescence microscopy

    NASA Astrophysics Data System (ADS)

    Bukara, Katarina; Jovanić, Svetlana; Drvenica, Ivana T.; Stančić, Ana; Ilić, Vesna; Rabasović, Mihailo D.; Pantelić, Dejan; Jelenković, Branislav; Bugarski, Branko; Krmpot, Aleksandar J.

    2017-02-01

    The present study describes utilization of two photon excitation fluorescence (2PE) microscopy for visualization of the hemoglobin in human and porcine erythrocytes and their empty membranes (i.e., ghosts). High-quality, label- and fixation-free visualization of hemoglobin was achieved at excitation wavelength 730 nm by detecting visible autofluorescence. Localization in the suspension and spatial distribution (i.e., mapping) of residual hemoglobin in erythrocyte ghosts has been resolved by 2PE. Prior to the 2PE mapping, the presence of residual hemoglobin in the bulk suspension of erythrocyte ghosts was confirmed by cyanmethemoglobin assay. 2PE analysis revealed that the distribution of hemoglobin in intact erythrocytes follows the cells' shape. Two types of erythrocytes, human and porcine, characterized with discocyte and echinocyte morphology, respectively, showed significant differences in hemoglobin distribution. The 2PE images have revealed that despite an extensive washing out procedure after gradual hypotonic hemolysis, a certain amount of hemoglobin localized on the intracellular side always remains bound to the membrane and cannot be eliminated. The obtained results open the possibility to use 2PE microscopy to examine hemoglobin distribution in erythrocytes and estimate the purity level of erythrocyte ghosts in biotechnological processes.

  5. Erythrocyte membrane sulfatide plays a crucial role in the adhesion of sickle erythrocytes to endothelium.

    PubMed

    Zhou, Z; Thiagarajan, P; Udden, M; Lòpez, J A; Guchhait, P

    2011-06-01

    Enhanced adhesion of sickle erythrocytes to the vascular endothelium and subendothelial matrix is fundamental to the development of vascular occlusion in sickle cell disease. Erythrocyte membrane sulfatide is implicated in the pathogenesis of vasoocclusive crises in sickle cell disease (SCD) patients. Because previous evidence linking sulfatide to cell adhesion has largely been circumstantial due to a lack of reagents that specifically target sulfatide, we used two sulfatide-specific strategies to address the role of erythrocyte membrane sulfatide in sickle cell adhesion to the vascular endothelium: a single-chain fragment variable chain (scFv) antibody against sulfatide as well as cerebroside sulfotransferase-deficient mice incapable of synthesising sulfatide. The sickle erythrocytes from mice and humans adhered at a greater extent and at higher shear stresses to activated endothelium than normal erythrocytes, and approximately 60% of the adhesion was prevented by the anti-sulfatide scFv. Similarly, the extent of adhesion of sulfatide-deficient erythrocytes was lower than normal erythrocytes. These findings suggest an important role for membrane sulfatide in sickle cell disease pathophysiology.

  6. Anarthria impairs subvocal counting.

    PubMed

    Cubelli, R; Nichelli, P; Pentore, R

    1993-12-01

    We studied subvocal counting in two pure anarthric patients. Analysis showed that they performed definitively worse than normal subjects free to articulate subvocally and their scores were in the lower bounds of the performances of subjects suppressing articulation. These results suggest that subvocal counting is impaired after anarthria.

  7. Complexities of Counting.

    ERIC Educational Resources Information Center

    Stake, Bernadine Evans

    This document focuses on one child's skip counting methods. The pupil, a second grade student at Steuben School, in Kankakee, Illinois, was interviewed as she made several attempts at counting twenty-five poker chips on a circular piece of paper. The interview was part of a larger study of "Children's Conceptions of Number and Numeral,"…

  8. Counting Sheep in Basque

    ERIC Educational Resources Information Center

    Araujo, Frank P.

    1975-01-01

    Demonstrates the interplay of a cognitive system, the Basque numerative system, and a behavioral one, counting sheep. The significant features of the Basque numerative system are analyzed; then it is shown how use of these features facilitates the counting of sheep on open ranges by Basque sheep farmers in California. (Author/RM)

  9. Averaging of TNTC counts.

    PubMed Central

    Haas, C N; Heller, B

    1988-01-01

    When plate count methods are used for microbial enumeration, if too-numerous-to-count results occur, they are commonly discarded. In this paper, a method for consideration of such results in computation of an average microbial density is developed, and its use is illustrated by example. PMID:3178211

  10. Counting Sheep in Basque

    ERIC Educational Resources Information Center

    Araujo, Frank P.

    1975-01-01

    Demonstrates the interplay of a cognitive system, the Basque numerative system, and a behavioral one, counting sheep. The significant features of the Basque numerative system are analyzed; then it is shown how use of these features facilitates the counting of sheep on open ranges by Basque sheep farmers in California. (Author/RM)

  11. Metabolism of acetylcholine in human erythrocytes

    SciTech Connect

    Chapman, E.S.

    1990-01-01

    In order to examine the possible role of erythrocyte acetylcholinesterase in the maintenance of membrane phospholipid content and membrane fluidity, experiments were performed to monitor the activity of the enzyme and follow the fate of one of its hydrolytic products, choline. Intact human erythrocytes were incubated with acetylcholine (choline methyl-{sup 14}C). The incubation resulted in the hydrolysis of acetylcholine to acetate and choline; the reaction was catalyzed by membrane acetylcholinesterase. The studies demonstrate the further metabolism of choline. Experiments were carried out to determine rate of hydrolysis of acetylcholine, uptake of choline, identification of intracellular metabolites of choline, and identification of radiolabeled membrane components. Erythrocytes at a 25% hematocrit were incubated in an isoosmotic bicarbonate buffer pH 7.4, containing glucose, adenosine, streptomycin and penicillin with 0.3 {mu}Ci of acetylcholine (choline methyl-{sup 14}C), for 24 hours. Aliquots of the erythrocyte suspension were taken throughout for analysis. Erythrocytes were washed free of excess substrate, lysed, and the hemolysate was extracted for choline and its metabolites. Blank samples containing incubation buffer and radiolabeled acetylcholine only, and erythrocyte hemolysate extracts were analyzed for choline content, the difference between blank samples and hemolysate extracts was the amount of choline originating from acetylcholine and attributable to acetylcholinesterase activity. The conversion of choline to {sup 14}C-betaine is noted after several minutes of incubation; at 30 minutes, more than 80% of {sup 14}C-choline is taken up and after several hours, detectable levels of radiolabeled S-adenosylmethionine were present in the hemolysate extract.

  12. Evaluation of erythrocyte and reticulocyte parameters as indicative of iron deficiency in patients with anemia of chronic disease

    PubMed Central

    Torino, Ana Beatriz Barbosa; Gilberti, Maria de Fátima Pererira; da Costa, Edvilson; de Lima, Gisélia Aparecida Freire; Grotto, Helena Zerlotti Wolf

    2015-01-01

    Objective The aim of this study was to evaluate the effectiveness of mature red cell and reticulocyte parameters to identify three conditions: iron deficiency anemia, anemia of chronic disease, and anemia of chronic disease associated with absolute iron deficiency. Methods Peripheral blood cells from 117 adult patients with anemia were classified according to iron status, inflammation, and hemoglobinopathies as: iron deficiency anemia (n = 42), anemia of chronic disease (n = 28), anemia of chronic disease associated with iron deficiency anemia (n = 22), and heterozygous β-thalassemia (n = 25). The percentage of microcytic erythrocytes, hypochromic erythrocytes, and the levels of hemoglobin in both reticulocytes and mature red cells were determined. Receiver operating characteristic analysis was used to evaluate the accuracy of the parameters in differentiating anemia. Results There was no difference between the groups of iron deficiency and anemia of chronic disease associated with absolute iron deficiency for any of the parameters. The percentage of hypochromic erythrocytes was the best parameter to identify absolute iron deficiency in patients with anemia of chronic disease (area under curve = 0.785; 95% confidence interval: 0.661–0.909 with sensitivity of 72.7%, and specificity of 70.4%; cut-off value 1.8%). The formula microcytic erythrocyte count minus hypochromic erythrocyte count was very accurate to differentiate iron deficiency anemia from heterozygous β-thalassemia (area under curve = 0.977; 95% confidence interval: 0.950–1.005 with a sensitivity of 96.2%, and specificity of 92.7%; cut-off value 13.8). Conclusion The erythrocyte and reticulocyte indices are moderately good to identify absolute iron deficiency in patients with anemia of chronic disease. PMID:25818816

  13. Transformation of human erythrocyte shape by endotoxic lipopolysaccharide.

    PubMed

    Warren, J R; Harris, A S; Wallas, C H

    1983-01-01

    Human erythrocytes were observed to undergo a discocyte to echinocyte to spheroechinocyte shape transformation during brief incubation with endotoxic lipopolysaccharide. It was concluded that lipopolysaccharide-membrane interactions alter the curvature of erythrocyte membranes.

  14. Transformation of Human Erythrocyte Shape by Endotoxic Lipopolysaccharide

    PubMed Central

    Warren, John R.; Harris, Alan S.; Wallas, Charles H.

    1983-01-01

    Human erythrocytes were observed to undergo a discocyte to echinocyte to spheroechinocyte shape transformation during brief incubation with endotoxic lipopolysaccharide. It was concluded that lipopolysaccharide-membrane interactions alter the curvature of erythrocyte membranes. Images PMID:6822423

  15. Distribution of diuretics and hypoglycemic sulfonylureas in rabbit erythrocytes.

    PubMed

    Yoshitomi, H; Kiko, S; Ikeda, K; Goto, S

    1983-03-01

    The distribution of three sulfonylureas and six diuretics in rabbit erythrocytes was studied in vitro at 37 degrees C. The drugs were taken up by the erythrocyte compartment, and distribution equilibrium was reached within 60 min of incubation. A distribution percentage in erythrocyte compartment was maintained at roughly constant value over the whole concentration range of drugs. Therefore, a linear relationship was established between total concentrations of drug in whole blood or erythrocyte suspension and in the erythrocyte compartment. Bovine serum albumin combined with the erythrocyte suspension appeared to reduce drug distribution in the erythrocyte compartment. Whole blood obtained from renal failure rabbits showed greater distribution of drug in the erythrocyte compartment compared with the whole blood of a normal rabbit. This might be due to a change in plasma protein binding ability related to the progress of renal failure.

  16. Erythrocytic and leukocytic responses to cadmium poisoning in freshwater fish, Puntius conchonius Ham

    SciTech Connect

    Gill, T.S.; Pant, J.C.

    1985-04-01

    Chronically sublethal concentrations of cadmium caused conspicuous hematological anomalies in the cyprinid fish, Puntius conchonius. Exposure to 0.63 and 0.84 mg/liter cadmium chloride (1/20 and 1/15 of 96-hr LC/sub 50/) induced morphological aberrations in mature erythrocytes including cytoplasmic vacuolation, hypochromia, deterioration of cellular membrane, basophilic stippling of cytoplasm, clumping of chromatin material and extrusion of nuclei, and schistocytosis. Anomalous basophils and monocytes were also encountered though less frequently. Decreased erythrocyte counts, hemoglobin and hematocrit values were also associated with chronic cadmium poisoning. The mean corpuscular hemoglobin and mean corpuscular volume increased (30 days) but mean corpuscular hemoglobin concentration showed no obvious change. A significant thrombocytopenia (90 days), elevated small lymphocyte and basophil populations, and a mild neutropenia were manifested in the cadmium-exposed fish. Large lymphocytes were not significantly affected.

  17. [Normobaric intermittent hypoxia and functional state of the erythrocyte pool].

    PubMed

    Dlusskaia, I G; Stepanov, V K; Radchenko, S N; Dvornikov, M V

    2004-01-01

    Functional state of the pool of erythrocytes was evaluated in ten essentially healthy male subjects before, during and in 2 months after a series of 15 exposures to normobaric intermittent hypoxia (NIH). The erythrocyte pool dynamics, hemoglobin content, low and highly resistive fractions of erythrocytes were analyzed using a modified acidic histogram technique. It was demonstrated that the erythrocyte pool was either in the state of destruction (concurrent to the NIH exposure) or ensuing persistent improvement of the functional characteristics under study.

  18. Increased caspase-3 immunoreactivity of erythrocytes in STZ diabetic rats.

    PubMed

    Fırat, Uğur; Kaya, Savaş; Cim, Abdullah; Büyükbayram, Hüseyin; Gökalp, Osman; Dal, Mehmet Sinan; Tamer, Mehmet Numan

    2012-01-01

    Eryptosis is a term to define apoptosis of erythrocytes. Oxidative stress and hyperglycemia, both of which exist in the diabetic intravascular environment, can trigger eryptosis of erythrocytes. In this experimental study, it is presented that the majority of erythrocytes shows caspase-3 immunoreactivity in streptozocin- (STZ)-induced diabetic rats. Besides that, caspase-3 positive erythrocytes are aggregated and attached to vascular endothelium. In conclusion, these results may start a debate that eryptosis could have a role in the diabetic complications.

  19. Erythrocyte sedimentation rate and C-reactive protein.

    PubMed

    Harrison, Michael

    2015-06-01

    C-reactive protein is a better indicator of inflammation than the erythrocyte sedimentation rate. It is more sensitive and responds more quickly to changes in the clinical situation. False negative and false positive results are more common when measuring the erythrocyte sedimentation rate. Renal disease, female sex and older age increase the erythrocyte sedimentation rate. The erythrocyte sedimentation rate has value in detecting low-grade bone infection, and in monitoring some patients with systemic lupus erythematosus.

  20. Elimination of Young Erythrocytes from Blood Circulation and Altered Erythropoietic Patterns during Paraquat Induced Anemic Phase in Mice

    PubMed Central

    Bhardwaj, Nitin; Saxena, Rajiv K.

    2014-01-01

    Paraquat a widely used herbicide causes a variety of toxic effects on humans and animals. The present study is focused on the interaction of paraquat with the mouse erythroid system. Administration of paraquat (10 mg/kg body weight i.p. on alternate days in C57Bl/6 mice) induced a significant fall in blood erythrocyte count on 7, 14, and 21 day time points but the erythrocyte count reverted back to normal by 28th day indicating the emergence of refractoriness to paraquat. A marked surge in the blood reticulocyte count was observed in paraquat treated mice that also subsided by 28th day. Young erythrocytes in circulation were randomly eliminated from blood circulation in paraquat treated mice and a significant elevation in the level of reactive oxygen species (ROS) was also observed maximally the erythrocytes of this age group. Cells representing various stages of erythroid differentiation in bone marrow and spleen were identified and enumerated flow cytometrically based on their expression of Ter119 and transferrin (CD71) receptor. Proliferative activity of erythroid cells, their relative proportion as well as their absolute numbers fell significantly in bone marrow of paraquat treated mice but all these parameters were significantly elevated in spleens of paraquat treated mice. These changes were essentially restricted to the cells belonging to the two earliest stages of erythroid differentiation. Taken together our results indicate that paraquat treatment causes a transient anemia in mice resulting from random elimination of young circulating erythrocytes as well as depressed erythropoietic activity in bone marrow. Spleen erythropoietic activity however was elevated in paraquat treated mice. PMID:24945144

  1. Elimination of young erythrocytes from blood circulation and altered erythropoietic patterns during paraquat induced anemic phase in mice.

    PubMed

    Bhardwaj, Nitin; Saxena, Rajiv K

    2014-01-01

    Paraquat a widely used herbicide causes a variety of toxic effects on humans and animals. The present study is focused on the interaction of paraquat with the mouse erythroid system. Administration of paraquat (10 mg/kg body weight i.p. on alternate days in C57Bl/6 mice) induced a significant fall in blood erythrocyte count on 7, 14, and 21 day time points but the erythrocyte count reverted back to normal by 28th day indicating the emergence of refractoriness to paraquat. A marked surge in the blood reticulocyte count was observed in paraquat treated mice that also subsided by 28th day. Young erythrocytes in circulation were randomly eliminated from blood circulation in paraquat treated mice and a significant elevation in the level of reactive oxygen species (ROS) was also observed maximally the erythrocytes of this age group. Cells representing various stages of erythroid differentiation in bone marrow and spleen were identified and enumerated flow cytometrically based on their expression of Ter119 and transferrin (CD71) receptor. Proliferative activity of erythroid cells, their relative proportion as well as their absolute numbers fell significantly in bone marrow of paraquat treated mice but all these parameters were significantly elevated in spleens of paraquat treated mice. These changes were essentially restricted to the cells belonging to the two earliest stages of erythroid differentiation. Taken together our results indicate that paraquat treatment causes a transient anemia in mice resulting from random elimination of young circulating erythrocytes as well as depressed erythropoietic activity in bone marrow. Spleen erythropoietic activity however was elevated in paraquat treated mice.

  2. An automated method for determining the cytoadhesion of Plasmodium falciparum-infected erythrocytes to immobilized cells.

    PubMed

    Hempel, Casper; Boisen, Ida M; Efunshile, Akinwale; Kurtzhals, Jørgen A L; Staalsø, Trine

    2015-03-14

    Plasmodium falciparum exports antigens to the surface of infected erythrocytes causing cytoadhesion to the host vasculature. This is central in malaria pathogenesis but in vitro studies of cytoadhesion rely mainly on manual counting methods. The current study aimed at developing an automated high-throughput method for this purpose utilizing the pseudoperoxidase activity of intra-erythrocytic haemoglobin. Chinese hamster ovary (CHO) cells were grown to confluence in chamber slides and microtiter plates. Cytoadhesion of co-cultured P. falciparum, selected for binding to CHO cells, was quantified by microscopy of Giemsa-stained chamber slides. In the automated assay, binding was quantified spectrophotometrically in microtiter plates after cell lysis using tetramethylbenzidine as peroxidase-catalysed substrate. The relevance of the method for binding studies was assessed using: i) binding of P. falciparum-infected erythrocytes to CHO cells over-expressing chondroitin sulfate A and ii) CHO cells transfected with CD36. Binding of infected erythrocytes including field isolates to primary endothelial cells was also performed. Data was analysed using linear regression and Bland-Altman plots. The manual and automated quantification showed strong, positive correlation (r(2) = 0.959, p <0.001) and with similar detection limit and precision. The automated assay showed the expected dose-dependent reduction in binding to CHO cells when blocking with soluble chondroitin sulfate A or anti-CD36 antibody. Quantification of binding to endothelial cells showed clear distinction between selected vs. non-selected parasite lines. Importantly, the assay was sufficiently sensitive to detect adhesion of field isolates to endothelial cells. The assay is simple and in a reproducible manner quantifies erythrocyte adhesion to several types of immobilized cells.

  3. Studies of the pathogenesis of anemia of inflammation: erythrocyte survival

    SciTech Connect

    Weiss, D.J.; Krehbiel, J.D.

    1983-10-01

    Erythrocyte survival was investigated in healthy cats and in cats with sterile abscesses. Erythrocyte survival time in cats with sterile abscesses was found to be significantly reduced. The erythrocyte destruction appeared to be the major factor in the early stages of anemia of inflammation.

  4. AUTOMATIC COUNTING APPARATUS

    DOEpatents

    Howell, W.D.

    1957-08-20

    An apparatus for automatically recording the results of counting operations on trains of electrical pulses is described. The disadvantages of prior devices utilizing the two common methods of obtaining the count rate are overcome by this apparatus; in the case of time controlled operation, the disclosed system automatically records amy information stored by the scaler but not transferred to the printer at the end of the predetermined time controlled operations and, in the case of count controlled operation, provision is made to prevent a weak sample from occupying the apparatus for an excessively long period of time.

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

    PubMed

    Hassan, Umer; Bashir, Rashid

    2014-10-01

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

  6. Erythrocyte survival in sheep exposed to ozone

    SciTech Connect

    Moore, G.S.; Calabrese, E.J.; Labato, F.J.

    1981-07-01

    Erythrocyte survival studies in the Dorset ewe using chromium 51 were performed. The purpose of the study was to determine if ozone exposure produces decreased cell survival which may be the result of premature erythrocyte aging. This strain of sheep has an erythrocyte glucose-6-phosphate dehydrogenase (G6PD) activity that is very low, being comparable to human A-variants with G6PD deficiency. Ozone exposure may produce hemolytic effects in G6PD deficients more readily than in erythrocytes with normal activity. A decrease in hematocrit was observed in the ozone exposed groups. With respect to red cell destruction, ozone does not appear to act immediately, but rather there appears to be a delayed effect. At 0.25 ppM ozone, the group reached the 50% remaining level an average of 1 day sooner than the control group. There was no significant difference between control and exposed groups at the 0.50 ppM and 0.70 ppM levels. Also, the results demonstrate a net decrease in hematocrit which is greater for 0.25 ppM ozone than any other exposure level. (RJC)

  7. Brucella melitensis invades murine erythrocytes during infection.

    PubMed

    Vitry, Marie-Alice; Hanot Mambres, Delphine; Deghelt, Michaël; Hack, Katrin; Machelart, Arnaud; Lhomme, Frédéric; Vanderwinden, Jean-Marie; Vermeersch, Marjorie; De Trez, Carl; Pérez-Morga, David; Letesson, Jean-Jacques; Muraille, Eric

    2014-09-01

    Brucella spp. are facultative intracellular Gram-negative coccobacilli responsible for brucellosis, a worldwide zoonosis. We observed that Brucella melitensis is able to persist for several weeks in the blood of intraperitoneally infected mice and that transferred blood at any time point tested is able to induce infection in naive recipient mice. Bacterial persistence in the blood is dramatically impaired by specific antibodies induced following Brucella vaccination. In contrast to Bartonella, the type IV secretion system and flagellar expression are not critically required for the persistence of Brucella in blood. ImageStream analysis of blood cells showed that following a brief extracellular phase, Brucella is associated mainly with the erythrocytes. Examination by confocal microscopy and transmission electron microscopy formally demonstrated that B. melitensis is able to invade erythrocytes in vivo. The bacteria do not seem to multiply in erythrocytes and are found free in the cytoplasm. Our results open up new areas for investigation and should serve in the development of novel strategies for the treatment or prophylaxis of brucellosis. Invasion of erythrocytes could potentially protect the bacterial cells from the host's immune response and hamper antibiotic treatment and suggests possible Brucella transmission by bloodsucking insects in nature.

  8. Electrophoretic mobilities of erythrocytes in various buffers

    NASA Technical Reports Server (NTRS)

    Plank, L. D.; Kunze, M. E.; Todd, P. W.

    1985-01-01

    The calibration of space flight equipment depends on a source of standard test particles, this test particle of choice is the fixed erythrocyte. Erythrocytes from different species have different electrophoretic mobilities. Electrophoretic mobility depends upon zeta potential, which, in turn depends upon ionic strength. Zeta potential decreases with increasing ionic strength, so cells have high electrophoretic mobility in space electrophoresis buffers than in typical physiological buffers. The electrophoretic mobilities of fixed human, rat, and rabbit erythrocytes in 0.145 M salt and buffers of varying ionic strength, temperature, and composition, to assess the effects of some of the unique combinations used in space buffers were characterized. Several effects were assessed: glycerol or DMSO (dimethylsulfoxide) were considered for use as cryoprotectants. The effect of these substances on erythrocyte electrophoretic mobility was examined. The choice of buffer depended upon cell mobility. Primary experiments with kidney cells established the choice of buffer and cryoprotectant. A nonstandard temperature of EPM in the suitable buffer was determined. A loss of ionic strength control occurs in the course of preparing columns for flight, the effects of small increases in ionic strength over the expected low values need to be evaluated.

  9. Stimulation of suicidal erythrocyte death by sulforaphane.

    PubMed

    Alzoubi, Kousi; Calabrò, Salvatrice; Faggio, Caterina; Lang, Florian

    2015-03-01

    Sulforaphane, an isothiocyanate from cruciferous vegetable, counteracts malignancy. The effect is at least in part due to the stimulation of suicidal death or apoptosis of tumour cells. Mechanisms invoked in sulforaphane-induced apoptosis include mitochondrial depolarization and altered gene expression. Despite the lack of mitochondria and nuclei, erythrocytes may, similar to apoptosis of nucleated cells, enter eryptosis, a suicidal cell death characterized by cell shrinkage and phosphatidylserine translocation to the erythrocyte surface. Stimulators of eryptosis include increase of cytosolic Ca(2+)-activity ([Ca(2+)]i). This study explored whether sulforaphane stimulates eryptosis. Cell volume was estimated from forward scatter, phosphatidylserine exposure at the cell surface from annexin V binding and [Ca(2+)]i from Fluo-3 fluorescence. A 48-hr treatment of human erythrocytes with sulforaphane (50-100 μM) significantly decreased forward scatter, significantly increased the percentage of annexin V binding cells and significantly increased [Ca(2+)]i. The effect of sulforaphane (100 μM) on annexin V binding was significantly blunted but not abrogated by the removal of extracellular Ca(2+). Sulforaphane (100 μM) significantly increased ceramide formation. In conclusion, sulforaphane stimulates suicidal erythrocyte death or eryptosis, an effect at least partially, but not exclusively, due to the stimulation of Ca(2+) entry and ceramide formation. © 2014 Nordic Association for the Publication of BCPT (former Nordic Pharmacological Society).

  10. OXALATE FORMATION FROM GLYOXAL IN ERYTHROCYTES

    PubMed Central

    Knight, John; Wood, Kyle D.; Lange, Jessica N.; Assimos, Dean G.; Holmes, Ross P.

    2015-01-01

    OBJECTIVES To determine whether glyoxal can be converted to oxalate in human erythrocytes. Glyoxal synthesis is elevated in diabetes, cardiovascular disease and other diseases with significant oxidative stress. Erythrocytes are a good model system for such studies as they lack intracellular organelles and have a simplified metabolism. METHODS Erythrocytes were isolated from healthy volunteers and incubated with varying concentrations of glyoxal for different amounts of time. Metabolic inhibitors were used to help characterize metabolic steps. The conversion of glyoxal to glycolate and oxalate in the incubation medium was determined by chromatographic techniques. RESULTS The bulk of the glyoxal was converted to glycolate but ~1% was converted to oxalate. Inclusion of the pro-oxidant, menadione, in the medium increased oxalate synthesis, and the inclusion of disulfiram, an inhibitor of aldehyde dehydrogenase activity, decreased oxalate synthesis. CONCLUSIONS The glyoxalase system, which utilizes glutathione as a cofactor, converts the majority of the glyoxal taken up by erythrocytes to glycolate but a small portion is converted to oxalate. A reduction in intracellular glutathione increases oxalate synthesis and a decrease in aldehyde dehydrogenase activity lowers oxalate synthesis and suggests that glyoxylate is an intermediate. Thus, oxidative stress in tissues could potentially increase oxalate synthesis. PMID:26546809

  11. Inhibition of suicidal erythrocyte death by xanthohumol.

    PubMed

    Qadri, Syed M; Mahmud, Hasan; Föller, Michael; Lang, Florian

    2009-08-26

    Xanthohumol is a proapoptotic hop-derived beer component with anticancer and antimicrobial activities. Similar to nucleated cells, erythrocytes may undergo suicidal cell death or eryptosis, which is triggered by oxidative stress (tert-butylhydroperoxide, TBOOH) or energy depletion (removal of glucose). The triggers increase cytosolic Ca(2+) concentration, leading to activation of Ca(2+)-sensitive K(+) channels with subsequent cell shrinkage and to cell membrane scrambling with subsequent phosphatidylserine exposure at the erythrocyte surface. Eryptotic cells are cleared from the circulating blood, leading to anemia, and may adhere to the vascular wall, thus impeding microcirculation. The present experiments explored whether xanthohumol influences eryptosis using flow cytometry. Exposure of human erythrocytes to 0.3 mM TBOOH or incubation in glucose-free solution significantly increased Fluo3 fluorescence (Ca(2+) concentration) as well as annexin V-binding (cell membrane scrambling) and decreased forward scatter (cell volume), effects significantly blunted by xanthohumol. In conclusion, xanthohumol is a potent inhibitor of suicidal erythrocyte death in vitro.

  12. Electrophoretic mobilities of erythrocytes in various buffers

    NASA Technical Reports Server (NTRS)

    Plank, L. D.; Kunze, M. E.; Todd, P. W.

    1985-01-01

    The calibration of space flight equipment depends on a source of standard test particles, this test particle of choice is the fixed erythrocyte. Erythrocytes from different species have different electrophoretic mobilities. Electrophoretic mobility depends upon zeta potential, which, in turn depends upon ionic strength. Zeta potential decreases with increasing ionic strength, so cells have high electrophoretic mobility in space electrophoresis buffers than in typical physiological buffers. The electrophoretic mobilities of fixed human, rat, and rabbit erythrocytes in 0.145 M salt and buffers of varying ionic strength, temperature, and composition, to assess the effects of some of the unique combinations used in space buffers were characterized. Several effects were assessed: glycerol or DMSO (dimethylsulfoxide) were considered for use as cryoprotectants. The effect of these substances on erythrocyte electrophoretic mobility was examined. The choice of buffer depended upon cell mobility. Primary experiments with kidney cells established the choice of buffer and cryoprotectant. A nonstandard temperature of EPM in the suitable buffer was determined. A loss of ionic strength control occurs in the course of preparing columns for flight, the effects of small increases in ionic strength over the expected low values need to be evaluated.

  13. Brucella melitensis Invades Murine Erythrocytes during Infection

    PubMed Central

    Vitry, Marie-Alice; Hanot Mambres, Delphine; Deghelt, Michaël; Hack, Katrin; Machelart, Arnaud; Lhomme, Frédéric; Vanderwinden, Jean-Marie; Vermeersch, Marjorie; De Trez, Carl; Pérez-Morga, David; Letesson, Jean-Jacques

    2014-01-01

    Brucella spp. are facultative intracellular Gram-negative coccobacilli responsible for brucellosis, a worldwide zoonosis. We observed that Brucella melitensis is able to persist for several weeks in the blood of intraperitoneally infected mice and that transferred blood at any time point tested is able to induce infection in naive recipient mice. Bacterial persistence in the blood is dramatically impaired by specific antibodies induced following Brucella vaccination. In contrast to Bartonella, the type IV secretion system and flagellar expression are not critically required for the persistence of Brucella in blood. ImageStream analysis of blood cells showed that following a brief extracellular phase, Brucella is associated mainly with the erythrocytes. Examination by confocal microscopy and transmission electron microscopy formally demonstrated that B. melitensis is able to invade erythrocytes in vivo. The bacteria do not seem to multiply in erythrocytes and are found free in the cytoplasm. Our results open up new areas for investigation and should serve in the development of novel strategies for the treatment or prophylaxis of brucellosis. Invasion of erythrocytes could potentially protect the bacterial cells from the host's immune response and hamper antibiotic treatment and suggests possible Brucella transmission by bloodsucking insects in nature. PMID:25001604

  14. Detergent induced lysis of erythrocytes in kwashiorkor.

    PubMed

    Rao, A; Onuora, C U; Cherian, A

    1987-09-15

    The effect of the non-ionic detergent Nonidet P40 on lysis of erythrocytes in children suffering from kwashiorkor was studied. The concentration of the detergent causing 50% haemolysis was significantly reduced in these patients. Detergent haemolysis was more sensitive than osmotic fragility (which was reduced). The abnormality was only slight in marasmic children.

  15. Glycophorin B is the erythrocyte receptor of Plasmodium falciparum erythrocyte-binding ligand, EBL-1

    PubMed Central

    Mayer, D. C. Ghislaine; Cofie, Joann; Jiang, Lubin; Hartl, Daniel L.; Tracy, Erin; Kabat, Juraj; Mendoza, Laurence H.; Miller, Louis H.

    2009-01-01

    In the war against Plasmodium, humans have evolved to eliminate or modify proteins on the erythrocyte surface that serve as receptors for parasite invasion, such as the Duffy blood group, a receptor for Plasmodium vivax, and the Gerbich-negative modification of glycophorin C for Plasmodium falciparum. In turn, the parasite counters with expansion and diversification of ligand families. The high degree of polymorphism in glycophorin B found in malaria-endemic regions suggests that it also may be a receptor for Plasmodium, but, to date, none has been identified. We provide evidence from erythrocyte-binding that glycophorin B is a receptor for the P. falciparum protein EBL-1, a member of the Duffy-binding-like erythrocyte-binding protein (DBL-EBP) receptor family. The erythrocyte-binding domain, region 2 of EBL-1, expressed on CHO-K1 cells, bound glycophorin B+ but not glycophorin B-null erythrocytes. In addition, glycophorin B+ but not glycophorin B-null erythrocytes adsorbed native EBL-1 from the P. falciparum culture supernatants. Interestingly, the Efe pygmies of the Ituri forest in the Democratic Republic of the Congo have the highest gene frequency of glycophorin B-null in the world, raising the possibility that the DBL-EBP family may have expanded in response to the high frequency of glycophorin B-null in the population. PMID:19279206

  16. Glycophorin B is the erythrocyte receptor of Plasmodium falciparum erythrocyte-binding ligand, EBL-1.

    PubMed

    Mayer, D C Ghislaine; Cofie, Joann; Jiang, Lubin; Hartl, Daniel L; Tracy, Erin; Kabat, Juraj; Mendoza, Laurence H; Miller, Louis H

    2009-03-31

    In the war against Plasmodium, humans have evolved to eliminate or modify proteins on the erythrocyte surface that serve as receptors for parasite invasion, such as the Duffy blood group, a receptor for Plasmodium vivax, and the Gerbich-negative modification of glycophorin C for Plasmodium falciparum. In turn, the parasite counters with expansion and diversification of ligand families. The high degree of polymorphism in glycophorin B found in malaria-endemic regions suggests that it also may be a receptor for Plasmodium, but, to date, none has been identified. We provide evidence from erythrocyte-binding that glycophorin B is a receptor for the P. falciparum protein EBL-1, a member of the Duffy-binding-like erythrocyte-binding protein (DBL-EBP) receptor family. The erythrocyte-binding domain, region 2 of EBL-1, expressed on CHO-K1 cells, bound glycophorin B(+) but not glycophorin B-null erythrocytes. In addition, glycophorin B(+) but not glycophorin B-null erythrocytes adsorbed native EBL-1 from the P. falciparum culture supernatants. Interestingly, the Efe pygmies of the Ituri forest in the Democratic Republic of the Congo have the highest gene frequency of glycophorin B-null in the world, raising the possibility that the DBL-EBP family may have expanded in response to the high frequency of glycophorin B-null in the population.

  17. Inventory count strategies.

    PubMed

    Springer, W H

    1996-02-01

    An important principle of accounting is that asset inventory needs to be correctly valued to ensure that the financial statements of the institution are accurate. Errors is recording the value of ending inventory in one fiscal year result in errors to published financial statements for that year as well as the subsequent fiscal year. Therefore, it is important that accurate physical counts be periodically taken. It is equally important that any system being used to generate inventory valuation, reordering or management reports be based on consistently accurate on-hand balances. At the foundation of conducting an accurate physical count of an inventory is a comprehensive understanding of the process coupled with a written plan. This article presents a guideline of the physical count processes involved in a traditional double-count approach.

  18. Counting Knights and Knaves

    ERIC Educational Resources Information Center

    Levin,Oscar; Roberts, Gerri M.

    2013-01-01

    To understand better some of the classic knights and knaves puzzles, we count them. Doing so reveals a surprising connection between puzzles and solutions, and highlights some beautiful combinatorial identities.

  19. Blood Count Tests

    MedlinePlus

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

  20. Understanding Blood Counts

    MedlinePlus

    ... Heart, Kidneys, Liver and Lung Function Infections Iron Overload Low Blood Counts Pain Disease- and Treatment-Related ... cancer research around the world and provides free information and support services. Privacy Policy Security Copyright Link ...

  1. Interactions of quantum dots with donor blood erythrocytes in vitro.

    PubMed

    Pleskova, S N; Pudovkina, E E; Mikheeva, E R; Gorshkova, E N

    2014-01-01

    The effects of quantum dots CdSe/ZnS-mercaptopropionic acid, (CdSe/CdZnS)ZnS-polyT, and CdSeCdSZnS/polyT/SiO2-NH2 on human erythrocytes were studied. The nanomaterials reduced signifi cantly the erythrocyte sedimentation rate and modified the erythrocyte membrane resistance to induced (acid and hypo-osmotic) hemolysis. Evaluation of the erythrocyte morphology by atomic force microscopy in the control and after exposure to quantum dots showed significant differences in erythrocyte size and changes in their morphology as a result of exposure to the nanomaterials.

  2. Neutron counting with cameras

    SciTech Connect

    Van Esch, Patrick; Crisanti, Marta; Mutti, Paolo

    2015-07-01

    A research project is presented in which we aim at counting individual neutrons with CCD-like cameras. We explore theoretically a technique that allows us to use imaging detectors as counting detectors at lower counting rates, and transits smoothly to continuous imaging at higher counting rates. As such, the hope is to combine the good background rejection properties of standard neutron counting detectors with the absence of dead time of integrating neutron imaging cameras as well as their very good spatial resolution. Compared to Xray detection, the essence of thermal neutron detection is the nuclear conversion reaction. The released energies involved are of the order of a few MeV, while X-ray detection releases energies of the order of the photon energy, which is in the 10 KeV range. Thanks to advances in camera technology which have resulted in increased quantum efficiency, lower noise, as well as increased frame rate up to 100 fps for CMOS-type cameras, this more than 100-fold higher available detection energy implies that the individual neutron detection light signal can be significantly above the noise level, as such allowing for discrimination and individual counting, which is hard to achieve with X-rays. The time scale of CMOS-type cameras doesn't allow one to consider time-of-flight measurements, but kinetic experiments in the 10 ms range are possible. The theory is next confronted to the first experimental results. (authors)

  3. Uric acid increases erythrocyte aggregation: Implications for cardiovascular disease.

    PubMed

    Sloop, Gregory D; Bialczak, Jessica K; Weidman, Joseph J; St Cyr, J A

    2016-10-05

    Uric acid may be a risk factor for atherosclerotic cardiovascular disease, although the data conflict and the mechanism by which it may cause cardiovascular disease is uncertain. This study was performed to test the hypothesis that uric acid, an anion at physiologic pH, can cause erythrocyte aggregation, which itself is associated with cardiovascular disease. Normal erythrocytes and erythrocytes with a positive direct antiglobulin test for surface IgG were incubated for 15 minutes in 14.8 mg/dL uric acid. Erythrocytes without added uric acid were used as controls. Erythrocytes were then examined microscopically for aggregation. Aggregates of up to 30 erythrocytes were noted when normal erythrocytes were incubated in uric acid. Larger aggregates were noted when erythrocytes with surface IgG were incubated in uric acid. Aggregation was negligible in controls. These data show that uric acid causes erythrocyte aggregation. The most likely mechanism is decreased erythrocyte zeta potential. Erythrocyte aggregates will increase blood viscosity at low shear rates and increase the risk of atherothrombosis. In this manner, hyperuricemia and decreased zeta potential may be risk factors for atherosclerotic cardiovascular disease.

  4. Lead transport and binding by human erythrocytes in vitro.

    PubMed

    Simons, T J

    1993-05-01

    Transport and binding of Pb2+ by human erythrocytes were examined for cell Pb contents in the 1-10 microM range, using the 203Pb isotope. Pb2+ crosses the erythrocyte membrane by the anion exchanger, and can also leave erythrocytes by a vanadate-sensitive pathway, identified with the Ca2+ pump. However, Pb2+ exit is very much less than expected from earlier experiments with resealed erythrocyte ghosts [Simons TJB (1988) J Physiol (Lond) 405:105-113] and the distribution of Pb2+ across the erythrocyte membrane is close to equilibrium. The high ratio of erythrocyte to plasma Pb seen in vivo appears to be due to the presence of a labile Pb(2+)-binding component present in erythrocyte cytoplasm.

  5. The Impact of Biophysical Properties of Erythrocytes on their Aggregation.

    PubMed

    Elblbesy, Mohamed A; Moustafa, Maisa E

    2017-06-01

    Erythrocytes aggregation takes places under low shear conditions or at stasis. All suggested mechanisms of erythrocytes aggregation indicated the importance role of fibrinogen and other blood proteins in enhanced erythrocyte aggregation. Recently a special attention is given to the cellular factors that may effect on erythrocytes aggregation. The present study inferred the effect of the cellular properties of erythrocytes on their aggregation. In the present study, aggregation index was calculated by a simple microscopic method. Correlations between erythrocytes aggregation index and mean cell volume, osmotic fragility, electrophoretic mobility, and magnetophoretic mobility were studied. The findings of this study indicated that the aggregation index is significatly correlated to mean cell volume, magnetophoretic mobility, osmotic fragility and electrophoretic mobility. Thus, It is concluded that cellular factors should be taken into consideration when studying the mechanism of erythrocytes aggregation.

  6. Study of erythrocyte membrane fluctuation using light scattering analysis

    NASA Astrophysics Data System (ADS)

    Lee, Hoyoon; Lee, Sangyun; Park, YongKeun; Shin, Sehyun

    2016-03-01

    It is commonly known that alteration of erythrocyte deformability lead to serious microcirculatory diseases such as retinopathy, nephropathy, etc. Various methods and technologies have been developed to diagnose such membrane properties of erythrocytes. In this study, we developed an innovative method to measure hemorheological characteristics of the erythrocyte membrane using a light scattering analysis with simplified optic setting and multi-cell analysis as well. Light scattering intensity through multiple erythrocytes and its power density spectrum were obtained. The results of light scattering analyses were compared in healthy control and artificially hardened sample which was treated with glutaraldehyde. These results were further compared with conventional assays to measure deformable property in hemorheology. We found that light scattering information would reflect the disturbance of membrane fluctuation in artificially damaged erythrocytes. Therefore, measuring fluctuation of erythrocyte membrane using light scattering signal could facilitate simple and precise diagnose of pathological state on erythrocyte as well as related complications.

  7. Effects of acute and chronic exercise on the osmotic stability of erythrocyte membrane of competitive swimmers

    PubMed Central

    2017-01-01

    This study aimed to evaluate the influence of acute and chronic exercise on erythrocyte membrane stability and various blood indices in a population consisting of five national-level male swimmers, over 18 weeks of training. The evaluations were made at the beginning and end of the 1st, 7th, 13th and 18th weeks, when volume and training intensity have changed. The effects manifested at the beginning of those weeks were considered due to chronic adaptations, while the effects observed at the end of the weeks were considered due to acute manifestations of the exercise load of that week. Acute changes resulting from the exercise comprised increases in creatine kinase activity (CK) and leukocyte count (Leu), and decrease in hematocrit (Ht) and mean corpuscular volume (MCV), at the end of the first week; increase in the activities of CK and lactate dehydrogenase (LDH), in the uric acid (UA) concentration and Leu count, at the end of the seventh week; increases in CK and LDH activities and in the mean corpuscular hemoglobin concentration (MCHC), at the end of the 13th week; and decrease in the value of the osmotic stability index 1/H50 and increases in the CK activity and platelets (Plt) count, at the end of the 18th week. Chronic changes due to training comprised increase in the values of 1/H50, CK, LDH, high-density lipoprotein cholesterol (HDL-C), low-density lipoprotein cholesterol (LDL-C), serum iron (Fe), MCV and Plt. Although acute training has resulted in decrease in the osmotic stability of erythrocytes, possibly associated with exacerbation of the oxidative processes during intense exercise, chronic training over 18 weeks resulted in increased osmotic stability of erythrocytes, possibly by modulation in the membrane cholesterol content by low and high density lipoproteins. PMID:28151958

  8. Effects of acute and chronic exercise on the osmotic stability of erythrocyte membrane of competitive swimmers.

    PubMed

    Paraiso, Lara Ferreira; Gonçalves-E-Oliveira, Ana Flávia Mayrink; Cunha, Lucas Moreira; de Almeida Neto, Omar Pereira; Pacheco, Adriana Garcia; Araújo, Karinne Beatriz Gonçalves; Garrote-Filho, Mário da Silva; Bernardino Neto, Morun; Penha-Silva, Nilson

    2017-01-01

    This study aimed to evaluate the influence of acute and chronic exercise on erythrocyte membrane stability and various blood indices in a population consisting of five national-level male swimmers, over 18 weeks of training. The evaluations were made at the beginning and end of the 1st, 7th, 13th and 18th weeks, when volume and training intensity have changed. The effects manifested at the beginning of those weeks were considered due to chronic adaptations, while the effects observed at the end of the weeks were considered due to acute manifestations of the exercise load of that week. Acute changes resulting from the exercise comprised increases in creatine kinase activity (CK) and leukocyte count (Leu), and decrease in hematocrit (Ht) and mean corpuscular volume (MCV), at the end of the first week; increase in the activities of CK and lactate dehydrogenase (LDH), in the uric acid (UA) concentration and Leu count, at the end of the seventh week; increases in CK and LDH activities and in the mean corpuscular hemoglobin concentration (MCHC), at the end of the 13th week; and decrease in the value of the osmotic stability index 1/H50 and increases in the CK activity and platelets (Plt) count, at the end of the 18th week. Chronic changes due to training comprised increase in the values of 1/H50, CK, LDH, high-density lipoprotein cholesterol (HDL-C), low-density lipoprotein cholesterol (LDL-C), serum iron (Fe), MCV and Plt. Although acute training has resulted in decrease in the osmotic stability of erythrocytes, possibly associated with exacerbation of the oxidative processes during intense exercise, chronic training over 18 weeks resulted in increased osmotic stability of erythrocytes, possibly by modulation in the membrane cholesterol content by low and high density lipoproteins.

  9. Mice deficient in the putative phospholipid flippase ATP11C exhibit altered erythrocyte shape, anemia, and reduced erythrocyte life span.

    PubMed

    Yabas, Mehmet; Coupland, Lucy A; Cromer, Deborah; Winterberg, Markus; Teoh, Narci C; D'Rozario, James; Kirk, Kiaran; Bröer, Stefan; Parish, Christopher R; Enders, Anselm

    2014-07-11

    Transmembrane lipid transporters are believed to establish and maintain phospholipid asymmetry in biological membranes; however, little is known about the in vivo function of the specific transporters involved. Here, we report that developing erythrocytes from mice lacking the putative phosphatidylserine flippase ATP11C showed a lower rate of PS translocation in vitro compared with erythrocytes from wild-type littermates. Furthermore, the mutant mice had an elevated percentage of phosphatidylserine-exposing mature erythrocytes in the periphery. Although erythrocyte development in ATP11C-deficient mice was normal, the mature erythrocytes had an abnormal shape (stomatocytosis), and the life span of mature erythrocytes was shortened relative to that in control littermates, resulting in anemia in the mutant mice. Thus, our findings uncover an essential role for ATP11C in erythrocyte morphology and survival and provide a new candidate for the rare inherited blood disorder stomatocytosis with uncompensated anemia. © 2014 by The American Society for Biochemistry and Molecular Biology, Inc.

  10. Fast counting electronics for neutron coincidence counting

    DOEpatents

    Swansen, James E.

    1987-01-01

    An amplifier-discriminator is tailored to output a very short pulse upon an above-threshold input from a detector which may be a .sup.3 He detector. The short pulse output is stretched and energizes a light emitting diode (LED) to provide a visual output of operation and pulse detection. The short pulse is further fed to a digital section for processing and possible ORing with other like generated pulses. Finally, the output (or ORed output ) is fed to a derandomizing buffer which converts the rapidly and randomly occurring pulses into synchronized and periodically spaced-apart pulses for the accurate counting thereof. Provision is also made for the internal and external disabling of each individual channel of amplifier-discriminators in an ORed plurality of same.

  11. Fast counting electronics for neutron coincidence counting

    DOEpatents

    Swansen, J.E.

    1985-03-05

    An amplifier-discriminator is tailored to output a very short pulse upon an above-threshold input from a detector which may be a /sup 3/He detector. The short pulse output is stretched and energizes a light emitting diode (LED) to provide a visual output of operation and pulse detection. The short pulse is further fed to a digital section for processing and possible ORing with other like generated pulses. Finally, the output (or ORed output) is fed to a derandomizing buffer which converts the rapidly and randomly occurring pulses into synchronized and periodically spaced-apart pulses for the accurate counting thereof. Provision is also made for the internal and external disabling of each individual channel of amplifier-discriminators in an ORed plurality of same.

  12. Whose interests count?

    PubMed

    Brudney, Daniel; Lantos, John D

    2014-10-01

    Whose interests should count and how should various interests be balanced at the pediatric patient's bedside? The interests of the child patient clearly count. Recently, however, many authors have argued that the family's interests also count. But how should we think about the interests of others? What does it mean to talk about "the family" in this context? Does it really just mean the interests of each individual family member? Or is the family itself a moral entity that has interests of its own independent of the interests of each of its members? Are such interests important only as they affect the patient's interest or also for their own sake? In this special supplement to Pediatrics, a group of pediatricians, philosophers, and lawyers grapple with these questions. They examine these issues from different angles and reach different conclusions. Jointly, they demonstrate the ethical importance and, above all, the ethical complexity of the family's role at the bedside.

  13. Interpretation of galaxy counts

    SciTech Connect

    Tinsely, B.M.

    1980-10-01

    New models are presented for the interpretation of recent counts of galaxies to 24th magnitude, and predictions are shown to 28th magnitude for future comparison with data from the Space Telescope. The results supersede earlier, more schematic models by the author. Tyson and Jarvis found in their counts a ''local'' density enhancement at 17th magnitude, on comparison with the earlier models; the excess is no longer significant when a more realistic mixture of galaxy colors is used. Bruzual and Kron's conclusion that Kron's counts show evidence for evolution at faint magnitudes is confirmed, and it is predicted that some 23d magnitude galaxies have redshifts greater than unity. These may include spheroidal systems, elliptical galaxies, and the bulges of early-type spirals and S0's, seen during their primeval rapid star formation.

  14. Erythrocyte choline concentrations and cluster headache.

    PubMed Central

    de Belleroche, J; Cook, G E; Das, I; Joseph, R; Tresidder, I; Rouse, S; Petty, R; Clifford Rose, F C

    1984-01-01

    Erythrocyte choline concentrations were measured in patients with cluster headache and age related control subjects. Concentrations were significantly reduced in the patients with headache both during a cluster period and between clusters, being 58% and 55% of the control value, respectively. After two weeks' treatment with lithium, choline concentrations in the patients with cluster headache increased to 78 times the control value (mean 369.2 mumol/l (3840 micrograms/100 ml) compared with 4.7 mumol/l (49 micrograms/100 ml]. The presence of depressed erythrocyte choline concentrations during and between cluster attacks indicates that this may be a predisposing condition which results in a cluster attack only when associated with a trigger factor. PMID:6419890

  15. Quantitation of CD55 and CD59 expression on reticulocytes and mature erythrocytes in paroxysmal nocturnal hemoglobinuria, aplastic anemia, and healthy control subjects.

    PubMed

    Kim, Yeongsic; Lim, Jihyang; Kim, Myungshin; Kim, Yonggoo; Lee, Jong-Wook; Han, Kyungja

    2010-01-01

    Since PNH occasionally results in bone marrow failure, it is difficult to differentiate PNH from AA with small numbers of CD(55) (-)CD(59) (-) erythrocytes. We quantified CD55 and CD59 molecules expressed on normal reticulocytes and mature erythrocytes of paroxysmal nocturnal hemoglobinuria (PNH), aplastic anemia (AA), and normal individuals in order to determine their usefulness for differentiation between PNH and AA. A total of 56 patients (AA 39 patients, PNH 17 patients) and 10 healthy volunteers were enrolled. Two-color flow cytometric analyses were conducted using thiazole orange, anti-CD55 and CD59 monoclonal antibodies to identify CD(55) (-)CD(59) (-) reticulocytes and mature erythrocytes. Mean fluorescence level of CD(55) (+) mature erythrocytes was lowest in the PNH patients (13.2 x 10(3) MESF), and was significantly lower than in normal controls (16.7 x 10(3) MESF, p <0.05), and highest in the AA subjects (22.8 x 10(3) MESF). Mean fluorescence level of CD(59) (+) mature erythrocytes was lowest in the PNH subjects (24.5 x 10(3) MESF), significantly less than in normal controls (39.0 x 10(3) MESF, p <0.05), and highest in the AA subjects (49.2 x 10(3) MESF). Total reticulocyte counts correlated strongly with the CD(55) (-)CD(59) (-) reticulocytes. Comparative quantitative analysis of CD55 and CD59 molecules expressed on normal erythrocytes may be useful in differentiating PNH from AA patients with small numbers of PNH-phenotype erythrocytes, and the total reticulocyte count may prove useful as a marker for PNH clone size.

  16. Green Hemoprotein of Erythrocytes: Methemoglobin Superoxide Transferase

    DTIC Science & Technology

    1988-01-01

    oxyhemoglobin containing GHP moglobin spectrum in Figure lB. However, appeared to convert to methemoglobin much bubbling air through the sample with GHP slower...hemo- + H20 + H + globin under most adverse conditions and preserves its dominant function as methemo- (3) GHP + R. + 02 - GHP(.O’) + R+ globin...in part by the United States 2953;1978. 7. Chee, P. P.; Lardy, H . A. Isolation from erythrocytesAir Force Office of Scientific Research. o re eorti

  17. Recovery of autologous erythrocytes in transfused patients.

    PubMed

    Wallas, C H; Tanley, P C; Gorrell, L P

    1980-01-01

    A microcapillary method utilizing phthalate esters or an ultracentrifuge method are both capable of separating autologous from homologous erythrocytes in polytransfused patients. The microcapillary technique which is readily adaptable to blood bank laboratories provides a previously unavailable method for defining blood group antigen typings in transfused patients. Such typings are of vital importance in the laboratory evaluation of transfused patients with multiple or weak blood group antibodies.

  18. Stimulation of Suicidal Erythrocyte Death by Tafenoquine.

    PubMed

    Al Mamun Bhuyan, Abdulla; Bissinger, Rosi; Stockinger, Katja; Lang, Florian

    2016-01-01

    The 8-aminoquinoline tafenoquine has been shown to be effective against Plasmodia, Leishmania and Trypanosoma. The substance is at least in part effective by triggering apoptosis of the parasites. Similar to apoptosis, erythrocytes may enter suicidal death or eryptosis, which is characterized by cell shrinkage and cell membrane scrambling with phosphatidylserine translocation to the erythrocyte surface. Signaling involved in the regulation of eryptosis include increase of cytosolic Ca2+ activity ([Ca2+]i), oxidative stress, ceramide, zVAD sensitive caspases, SB203580 sensitive p38 kinase, staurosporine sensitive protein kinase C as well as D4476 sensitive casein kinase. The present study explored, whether tafenoquine induces eryptosis and aimed to possibly identify cellular mechanisms involved. Flow cytometry was employed to estimate phosphatidylserine exposure at the cell surface from annexin-V-binding, cell volume from forward scatter, [Ca2+]i from Fluo3-fluorescence, ROS formation from 2',7'-dichlorodihydrofluorescein diacetate (DCFDA) dependent fluorescence, and ceramide abundance utilizing specific antibodies. A 48 hours exposure of human erythrocytes to tafenoquine (500 ng/ml) significantly increased the percentage of annexin-V-binding cells, significantly decreased forward scatter, significantly increased Fluo3-fluorescence, and significantly increased DCFDA fluorescence. Tafenoquine did not significantly modify ceramide abundance. The effect of tafenoquine on annexin-V-binding was significantly blunted but not abolished by removal of extracellular Ca2+. The effect of tafenoquine on annexin-V-binding was not significantly blunted by zVAD (10 µM), SB203580 (2 µM) or staurosporine (1 µM). The effect of tafenoquine on annexin-V-binding was significantly blunted but not abolished by D4476 (10 µM). Tafenoquine triggers cell shrinkage and phospholipid scrambling of the erythrocyte cell membrane, an effect at least in part due to stimulation of Ca2+ entry

  19. Determinants of Erythrocyte Hydration In Current Opinion in Hematology

    PubMed Central

    Rinehart, Jesse; Gulcicek, Erol E.; Joiner, Clinton H.; Lifton, Richard P.; Gallagher, Patrick G.

    2012-01-01

    Purpose of Review Maintenance of cellular water and solute homeostasis is critical for survival of the erythrocyte. Inherited or acquired disorders that perturb this homeostasis jeopardize the erythrocyte, leading to its premature destruction. This report reviews recent progress in our understanding the determinants of erythrocyte hydration and its related disorders. Recent Findings The molecular and genetic bases of primary disorders of erythrocyte hydration are poorly understood. Recent studies have implicated roles for the anion transporter, SLC4A1, and the Rh-associated glycoprotein, RhAG. The most common secondary disorder associated with perturbed hydration of the erythrocyte is sickle cell disease, where dehydration contributes to disease pathology and clinical complications. Advances in understanding the mechanisms regulating erythrocyte solute and water content, particularly associated with KCl cotransport and Gardos channel activation, have revealed novel signaling mechanisms controlling erythrocyte hydration. These signaling pathways may provide innovative strategies to prevent erythrocyte dehydration in sickle cell disease. Summary Clinical, translational and biologic studies all contribute to our knowledge of erythrocyte hydration. Understanding the mechanisms controlling erythrocyte water and solute homeostasis will serve as a paradigm for other cells and may reveal new therapeutic targets for disease prevention and treatment. PMID:20182354

  20. New insights on hereditary erythrocyte membrane defects

    PubMed Central

    Andolfo, Immacolata; Russo, Roberta; Gambale, Antonella; Iolascon, Achille

    2016-01-01

    After the first proposed model of the red blood cell membrane skeleton 36 years ago, several additional proteins have been discovered during the intervening years, and their relationship with the pathogenesis of the related disorders have been somewhat defined. The knowledge of erythrocyte membrane structure is important because it represents the model for spectrin-based membrane skeletons in all cells and because defects in its structure underlie multiple hemolytic anemias. This review summarizes the main features of erythrocyte membrane disorders, dividing them into structural and altered permeability defects, focusing particularly on the most recent advances. New proteins involved in alterations of the red blood cell membrane permeability were recently described. The mechanoreceptor PIEZO1 is the largest ion channel identified to date, the fundamental regulator of erythrocyte volume homeostasis. Missense, gain-of-function mutations in the PIEZO1 gene have been identified in several families as causative of dehydrated hereditary stomatocytosis or xerocytosis. Similarly, the KCNN4 gene, codifying the so called Gardos channel, has been recently identified as a second causative gene of hereditary xerocytosis. Finally, ABCB6 missense mutations were identified in different pedigrees of familial pseudohyperkalemia. New genomic technologies have improved the quality and reduced the time of diagnosis of these diseases. Moreover, they are essential for the identification of the new causative genes. However, many questions remain to solve, and are currently objects of intensive studies. PMID:27756835

  1. Erythrocyte phosphatidylserine exposure in β-thalassemia.

    PubMed

    Ibrahim, Hamdy A; Fouda, Manal I; Yahya, Raida S; Abousamra, Nashwa K; Abd Elazim, Rania A

    2014-06-01

    [ABS]Phospholipid asymmetry is well maintained in erythrocyte (RBC) membranes with phosphatidylserine (PS) exclusively present in the inner leaflet. Eryptosis, the suicidal death of RBCs, is characterized by cell shrinkage, membrane blebbing, and cell membrane phospholipids scrambling with PS exposure at the cell surface. Erythrocytes exposing PS are recognized, bound, engulfed, and degraded by macrophages. Eryptosis thus fosters clearance of affected RBCs from circulating blood, which may aggravate anemia in pathological conditions. Thalassemia patients are more sensitive to the eryptotic depletion and osmotic shock which may affect RBC membrane phospholipid asymmetry. We aimed in this work to determine the RBC PS exposure in splenectomized and nonsplenectomized β-thalassemia major (β-TM) patients and correlate it with the clinical presentation and laboratory data. RBCs were stained for annexin V to detect phosphatidylserine (PS) exposure in 46 β-TM patients (27 splenectomized and 19 nonsplenectomized) compared to 17 healthy subjects as a control group. We observed a significant increase in RBC PS exposure in β-TM patients compared to control group (P = .0001). Erythrocyte PS exposure was significantly higher in splenectomized β-TM patients compared with nonsplenectomized β-TM patients (P = .001). No correlation was found between RBC PS exposure and clinical or hematological data of β-TM patients, but there was a positive correlation between RBC PS exposure and ferritin level in β-TM patients have higher levels of RBC PS exposure, and splenectomy was shown to aggravate RBC PS exposure without aggravation of anemia.

  2. Stabilization of Erythrocyte Membranes by Polyamines

    NASA Astrophysics Data System (ADS)

    Ballas, Samir K.; Mohandas, Narla; Marton, Laurence J.; Shohet, Stephen B.

    1983-04-01

    Using a laser diffraction technique, we have studied the effects of putrescine, spermidine, and spermine, the three physiologic polyamines, on the deformability and mechanical stability of human erythrocyte membranes. Ghosts resealed with polyamines were subjected to high fluid shear stress in an ektacytometer. All polyamines increased the membrane shear modulus (decreased deformability) in a concentration- and time-dependent manner. The order of effectiveness was spermine > spermidine > putrescine. At 10 μ M, spermine appreciably decreased membrane deformability. For the measurement of membrane mechanical stability, resealed ghosts were subjected to constant high shear stress in the ektacytometer and deformability was continuously recorded as the deformable ghosts fragmented into rigid spherical vesicles. Polyamines, especially spermine, caused a noticeable increase in the t1/2 for fragmentation. These effects could not be ascribed to proteolysis or Ca2+-induced transglutamination. That the effects of polyamines were specific and not simply due to their positive charge was demonstrated by the finding that Ca2+ and Mg2+ destabilized the erythrocyte membrane as evidenced by decreasing the t1/2 for fragmentation. Extracellular polyamines were not effective except under conditions that caused significant accumulation inside the cell. The data indicate that intracellular physiologic polyamines, especially spermine, decrease erythrocyte membrane deformability and stabilize the membrane skeleton, making it more resistant to fragmentation.

  3. New insights on hereditary erythrocyte membrane defects.

    PubMed

    Andolfo, Immacolata; Russo, Roberta; Gambale, Antonella; Iolascon, Achille

    2016-11-01

    After the first proposed model of the red blood cell membrane skeleton 36 years ago, several additional proteins have been discovered during the intervening years, and their relationship with the pathogenesis of the related disorders have been somewhat defined. The knowledge of erythrocyte membrane structure is important because it represents the model for spectrin-based membrane skeletons in all cells and because defects in its structure underlie multiple hemolytic anemias. This review summarizes the main features of erythrocyte membrane disorders, dividing them into structural and altered permeability defects, focusing particularly on the most recent advances. New proteins involved in alterations of the red blood cell membrane permeability were recently described. The mechanoreceptor PIEZO1 is the largest ion channel identified to date, the fundamental regulator of erythrocyte volume homeostasis. Missense, gain-of-function mutations in the PIEZO1 gene have been identified in several families as causative of dehydrated hereditary stomatocytosis or xerocytosis. Similarly, the KCNN4 gene, codifying the so called Gardos channel, has been recently identified as a second causative gene of hereditary xerocytosis. Finally, ABCB6 missense mutations were identified in different pedigrees of familial pseudohyperkalemia. New genomic technologies have improved the quality and reduced the time of diagnosis of these diseases. Moreover, they are essential for the identification of the new causative genes. However, many questions remain to solve, and are currently objects of intensive studies. Copyright© Ferrata Storti Foundation.

  4. Erythrocyte autoantibodies, autoimmune haemolysis, and carcinoma.

    PubMed Central

    Sokol, R J; Booker, D J; Stamps, R

    1994-01-01

    AIMS--To examine a large series of patients in whom both red cell autoantibodies and carcinoma are present; and to determine whether this rare occurrence is a true association or a chance event. METHODS--The laboratory records of 160 patients (76 men, 84 women; mean age 68 years) with erythrocyte autoantibodies and confirmed carcinoma were examined for site of tumour origin and clinical and immunohematological findings. To test whether the concomitant occurrence of autoantibodies and carcinoma was fortuitous, data on total population and carcinoma incidence were included in a chi 2 analysis. RESULTS--The association was significant (chi 2 = 97.5, p < 0.0005); erythrocyte autoantibodies and carcinoma were found together 12-13 times more often than expected from their relative frequencies. Autoantibodies occurred with a variety of carcinomas, particularly those of breast, lung, colon, rectum, and prostate; this largely reflected tumour incidence. Adenocarcinoma, squamous, anaplastic, and transitional cell types were all represented. Warm, cold, and mixed autoantibodies were not associated with particular tumour sites or histology. Eighty six patients had haemolysis of varying severity, 37 had metastatic disease, and 28 died within a few months of presentation. CONCLUSIONS--The presence of erythrocyte autoantibodies and carcinoma in the same patient is a true association and probably reflects a fundamental disturbance in immune homeostasis. It tends to occur with a large tumour mass and metastatic disease, and generally indicates a poor prognosis. PMID:8027372

  5. Accounting for What Counts

    ERIC Educational Resources Information Center

    Milner, Joseph O.; Ferran, Joan E.; Martin, Katharine Y.

    2003-01-01

    No Child Left Behind legislation makes it clear that outside evaluators determine what gets taught in the classroom. It is important to ensure they measure what truly counts in school. This fact is poignantly and sadly true for the under funded, poorly resourced, "low performing" schools that may be hammered by administration accountants…

  6. Counting “exotics”

    Treesearch

    Qinfeng Guo

    2011-01-01

    An introduced or exotic species is commonly defined as an organism accidentally or intentionally introduced to a new location by human activity (Williamson 1996; Richardson et al. 2000; Guo and Ricklefs 2010). However, the counting of exotics is often inconsistent. For example, in the US, previously published plant richness data for each state are only those either...

  7. What Counts as Evidence?

    ERIC Educational Resources Information Center

    Dougherty Stahl, Katherine A.

    2014-01-01

    Each disciplinary community has its own criteria for determining what counts as evidence of knowledge in their academic field. The criteria influence the ways that a community's knowledge is created, communicated, and evaluated. Situating reading, writing, and language instruction within the content areas enables teachers to explicitly…

  8. What Counts as Evidence?

    ERIC Educational Resources Information Center

    Dougherty Stahl, Katherine A.

    2014-01-01

    Each disciplinary community has its own criteria for determining what counts as evidence of knowledge in their academic field. The criteria influence the ways that a community's knowledge is created, communicated, and evaluated. Situating reading, writing, and language instruction within the content areas enables teachers to explicitly…

  9. LOW ENERGY COUNTING CHAMBERS

    DOEpatents

    Hayes, P.M.

    1960-02-16

    A beta particle counter adapted to use an end window made of polyethylene terephthalate was designed. The extreme thinness of the film results in a correspondingly high transmission of incident low-energy beta particles by the window. As a consequence, the counting efficiency of the present counter is over 40% greater than counters using conventional mica end windows.

  10. WY Kids Count.

    ERIC Educational Resources Information Center

    Wyoming Kids Count, Cheyenne.

    This WY Kids Count brochure uses the metaphor of children's building blocks to present information on the current well-being of Wyoming children and to advocate enhancing the lives of young children. Each block (i.e., each develop the brochure) presents concerns in a separate area: (1) poverty, highlighting the number of children living in…

  11. Spectral Markers of Erythrocytes on Solid Substrate

    NASA Astrophysics Data System (ADS)

    Paiziev, Adkhamjon A.; Krakhmalev, V. A.

    Proposed in previous paper [1,2] the new nondestructive method of optical microscopy allows to examine the structures of living cells (human erythrocytes) in their natural colors without its staining by using a specially designed substrate for deposition of biological sample and observing a native blood smears in reflected light. Color interference contrast image is achieved due to special condition of experiment is connected with chose of angle of incidental light, wave length of light of reflected ray, chemical composition of sample, thickness of sample, refractive index of sample, refractive index of substrate, chemical composition of substrate [1,2]. We can identify chemical compounds of erythrocytes after calibration color scale by alternative methods. For comparison we used Synchrotron Radiation based Fourier Transformed Infrared (SR-FTIR) microspectroscopy. By focusing of infrared beam of FTIR microscope on cell surface we can screen and distinguish difference erythrocytes by its color. For example on Fig. 49.1 we can see two neighbored erythrocytes where one of them have red color (point 1) and other-green (point 5). To identify their spectral markers we measured IR absorption spectra of cells at different points (1,2,3,4 and 5). Intermediated area (points 3 and 4) correspond to substrate spectra (silicon substrate) and their spectra are same. The peaks at 2,850 and 2,920 cm-1 correspond mainly to the CH2 stretching modes of the methylene chains in membrane lipids. At 1,650 cm-1 the amide I band is observed, which results, principally, from the n(CO) stretching vibrations of the protein amide bonds; the amide II band, near 1,550 cm-1, is a combination of the d(N-H) bending and n(C-N) stretching vibrations of the amide bonds. The peaks at 2,850 and 2,920 cm-1 correspond mainly to the CH2 stretching modes of the methylene chains in membrane lipids [3. The intensities of the absorption bands at 2,920 and 2,850 cm-1 in green erythrocyte (point 5) were also

  12. Common toad Rhinella arenarum (Hensel, 1867) and its importance in assessing environmental health: test of micronuclei and nuclear abnormalities in erythrocytes.

    PubMed

    Pollo, Favio E; Bionda, Clarisa L; Salinas, Zulma A; Salas, Nancy E; Martino, Adolfo L

    2015-09-01

    Anthropogenic activities may generate significant changes in the integrity of aquatic ecosystems, so long-term monitoring of populations that inhabit them is crucial. Counting micronucleated erythrocytes (MN) and erythrocytic nuclear abnormalities (ENA) in peripheral blood is a widely used method for detecting chromosomal damage due to chemical agents in the water. We analyzed MN and ENA frequency in blood obtained from the common toad Rhinella arenarum populations in sites with different degrees of environmental degradation. The results of this study indicate that there is an association between the frequency of micronuclei and nuclear abnormalities and the degree of environmental alteration recorded for the sites studied.

  13. Carbohydrate counting of food.

    PubMed

    Hegar, Karin; Heiber, Stefanie; Brändle, Michael; Christ, Emanuel; Keller, Ulrich

    2011-07-07

    Carbohydrate counting is a principal strategy in nutritional management of type 1 diabetes. The Nutri-Learn buffet (NLB) is a new computer-based tool for patient instruction in carbohydrate counting. It is based on food dummies made of plastic equipped with a microchip containing relevant food content data. The tool enables the dietician to assess the patient's food counting abilities and the patient to learn in a hands-on interactive manner to estimate food contents such as carbohydrate content. Multicentre randomised controlled trial in 134 patients with type 1 diabetes comparing the use of the Nutri-Learn buffet in determining and improving ability to estimate the carbohydrate content of food with the use of conventional counselling tools (i.e. pictures and tables). The NLB group showed significantly better carbohydrate estimation values than the control group. In particular, there was a significant improvement in estimation of starches, fruits and sweets. The NLB was preferred by patients and dieticians in that rating of carbohydrate was closer to reality than the use of conventional tools, and since the tool has a play element, is interactive and adjustable, and can be used with only minimal knowledge of a specific language. Adjustment of preprandial insulin doses to the amounts of dietary carbohydrates ingested during the subsequent meal resulted in improved metabolic control in previous studies. The present study demonstrated that the new tool (Nutri-Learn buffet) improved teaching and learning of carbohydrate counting. In addition, it allowed an objective assessment of the carbohydrate counting skills of patients by the dietician. The findings therefore suggest that the tool is helpful in nutritional counselling of patients with diabetes mellitus.

  14. Further studies on osmotic resistance of nucleated erythrocytes: observations with pigeon, peafowl, lizard and toad erythrocytes during changes in temperature and pH.

    PubMed

    Oyewale, J O

    1994-02-01

    The osmotic resistance of pigeon, peafowl, lizard and toad erythrocytes at different temperatures and pH was studied. Erythrocytes from female pigeons showed greater osmotic resistance than those from males, but no sex difference appeared with erythrocytes from peafowls. Pigeon erythrocytes were more resistant and the red blood cell, packed cell volume and haemoglobin values were higher than those in peafowls. Although no significant differences appeared in their haematological values, erythrocytes from the lizard were more resistant than erythrocytes from the toad. At higher temperature, the osmotic resistance of pigeon, lizard and toad erythrocytes increased, while that of peafowl erythrocytes decreased. The resistance of toad erythrocytes decreased in acidic and alkaline solutions, but that of peafowl erythrocytes increased in both solutions. However, with pigeon and lizard erythrocytes, the resistance was unaltered in alkaline solution and decreased in acidic solution.

  15. Metabolomic analysis of normal and sickle cell erythrocytes.

    PubMed

    Darghouth, D; Koehl, B; Junot, C; Roméo, P-H

    2010-09-01

    Metabolic signatures of specialized circulating hematopoietic cells in physiological or human hematological diseases start to be described. We use a simple and highly reproductive extraction method of erythrocytes metabolites coupled with a liquid chromatography-mass spectrometry based metabolites profiling method to determine metabolomes of normal and sickle cell erythrocytes. Sickle cell erythrocytes and normal erythrocytes metabolomes display major differences in glycolysis, in glutathione, in ascorbate metabolisms and in metabolites associated to membranes turnover. In addition, the amounts of metabolites derived from urea cycle and NO metabolism that partly take place within erythrocyte were different between normal and sickle cell erythrocytes. These results show that metabolic profiling of red blood cell diseases can now be determined and might indicate new biomarkers that can be used for the follow-up of sickle cell patients.

  16. The counting of native blood cells by digital microscopy

    NASA Astrophysics Data System (ADS)

    Torbin, S. O.; Doubrovski, V. A.; Zabenkov, I. V.; Tsareva, O. E.

    2017-03-01

    An algorithm for photographic images processing of blood samples in its native state was developed to determine the concentration of erythrocytes, leukocytes and platelets without individual separate preparation of cells' samples. Special "photo templates" were suggested to use in order to identify red blood cells. The effect of "highlighting" of leukocytes, which was found by authors, was used to increase the accuracy of this type of cells counting. Finally to raise the resolution of platelets from leukocytes the areas of their photo images were used, but not their sizes. It is shown that the accuracy of cells counting for native blood samples may be comparable with the accuracy of similar studies for smears. At the same time the proposed native blood analysis simplifies greatly the procedure of sample preparation in comparison to smear, permits to move from the detection of blood cells ratio to the determination of their concentrations in the sample.

  17. [Electronic platelet counting with particular reference to thrombocytopenias (author's transl)].

    PubMed

    Kuse, R; Burmeister, H; Hausmann, K

    1977-09-29

    Platelet counts in platelet-rich plasma without hematocrit dependent correction were performed by following rapid and simple steps: 1. pre-dilution of 20 microliter of whole blood by an isotonic solution 1:25; 2. stabilized low-speed centrifugation with 55 g for 5 minutes; 3. final dilution 1 : 5000; 4. enumeration by use of a TOA platelet counter PL-100 which has been technically improved in comparison to similar machines. Erroneously high results were obtained after a too short or too low centrifugation. As reason for this artifical small pulses due to disturbances of the flow patterns around the aperture (so-called vortex-effect) can be assumed having been caused by large-volumed erythrocytes and leukocytes in the suspension. The routinely used procedure was reliable for all platelet ranges, especially in thrombocytopenias between 100 X 10(9)/l and 25 X 10(9)l. In lower ranges comparisons with visual counts are essential.

  18. Uric acid protects erythrocytes from ozone-induced changes.

    PubMed

    Meadows, J; Smith, R C

    1987-08-01

    Uric acid effectively reduced hemolysis and methemoglobin formation in bovine and swine erythrocytes bubbled with ozone in vitro. In bovine erythrocytes, formation of thiobarbituric acid-reactive material was inhibited by uric acid, but there was little immediate protection for the swine cells. Antioxidant protection was due to preferential degradation of the uric acid by ozone. These results provide evidence to support the hypothesis that in plasma, uric acid can provide antioxidant protection for erythrocytes.

  19. Effect of thiol drugs on tert-butyl hydroperoxide induced luminol chemiluminescence in human erythrocytes, erythrocyte lysate, and erythrocyte membranes.

    PubMed

    Sajewicz, Waldemar

    2010-07-30

    The paper investigates the effect of thiol drugs (RSH) under oxidative stress condition using luminol-enhanced chemiluminescence technique. The examinations included N-acetylcysteine (NAC), N-acetylpenicillamine (NAP), penicillamine (PEN), mesna (MES), and tiopronin (TPR). The model systems contained isolated human erythrocytes (RBC), erythrocyte lysates (LYS) or erythrocyte membranes (MEM) exposed to tert-butyl hydroperoxide (t-BuOOH). Under the influence of RSH, a bimodal character of some experimental chemiluminescence curves was observed and the kinetic solution was considered as the sum of two logistic-exponential processes. These chemiluminescence changes probably reflected two connected processes--scavenging by RSH of the t-BuOOH-induced free radicals and simultaneous generation of thiol-derived secondary free radicals. Individual differences in thiols interaction showed a multivariate set of the kinetic curve descriptors. The Principal Component Analysis (PCA) well distinguished subsets of RSH influence in systems with RBC or LYS. Generally, the action of NAC was exclusively pro-oxidant in both systems, with RBC and LYS. The behaviour of MES or NAP in these systems was also pro-oxidant but many times less prominent than NAC. Under the influence of TPR a dramatic switch in the anti-oxidant effect was observed in system with RBC to very pro-oxidant effect in LYS. The influence of PEN was analogical to TPR but very weak. This experimental model together with kinetic solution of the unique bimodal chemiluminescence curves, and PCA, supply new insights to the dual (anti- and pro-oxidant) effects of thiol drugs under oxidative stress condition. (c) 2010 Elsevier Ireland Ltd. All rights reserved.

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

    PubMed

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

    2017-02-01

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

  1. Accumulation of Paprika Carotenoids in Human Plasma and Erythrocytes.

    PubMed

    Nishino, Azusa; Ichihara, Takashi; Takaha, Takeshi; Kuriki, Takashi; Nihei, Hideko; Kawamoto, Kazuhisa; Yasui, Hiroyuki; Maoka, Takashi

    2015-01-01

    The accumulation (incorporation) of paprika carotenoid in human plasma and erythrocytes was investigated. A paprika carotenoid supplement (14 mg/day) was ingested for 4 weeks by 5 young healthy volunteers (3 men and 2 women). After 2 weeks of carotenoid ingestion, the carotenoid levels in plasma and erythrocytes increased by 1.2-fold and 2.2-fold, respectively. Characteristic carotenoids found in paprika (capsanthin, cucurbitaxanthin A, and cryptocapsin) were detected in both plasma and erythrocytes. An oxidative metabolite of capsanthin (capsanthone) was also found in both plasma and erythrocytes.

  2. [Functional state feature of erythrocytes in healthy term newborn infants].

    PubMed

    Evsiukova, I I; Iakushenko, N S; Andreeva, A A; Shevel'kova, A A; Kolesova, T A; Katiukhin, L N; Dobrylko, I A; Mandukshev, I V

    2014-01-01

    Hematological parameters and functional status of erythrocytes were studied by the osmotic and ammonium loads in healthy newborns and in adults. Mean erythrocyte volume of newborns more than in adults. Significant difference index of osmotic fragility of neonates were observed in the transition from swelling to hemolysis. Kinetic of erythrocyte's hemolysis in the ammonium load was studied by low-angle light scattering (LaSca-analyzer). The percentage of erythrocyte hemolysis is lower and the velocity of hemolysis is 2.5 times slower in newborns than in adults.

  3. Preparation of Stable Sensitized Erythrocytes for Detection of Chlamydial Antibodies

    PubMed Central

    Lewis, Vester J.; Engelman, Helen M.; Thacker, W. Lanier

    1975-01-01

    Sheep erythrocytes were treated with glutaraldehyde before sensitization for the indirect hemagglutination test to assay chlamydial antibodies. This treatment markedly increased stability during storage. PMID:809464

  4. Antioxidant effect of lutein towards phospholipid hydroperoxidation in human erythrocytes.

    PubMed

    Nakagawa, Kiyotaka; Kiko, Takehiro; Hatade, Keijiro; Sookwong, Phumon; Arai, Hiroyuki; Miyazawa, Teruo

    2009-11-01

    Peroxidised phospholipid-mediated cytotoxity is involved in the pathophysiology of many diseases; for example, phospholipid hydroperoxides (PLOOH) are abnormally increased in erythrocytes of dementia patients. Dietary carotenoids (especially xanthophylls, polar carotenoids such as lutein) have gained attention as potent inhibitors against erythrocyte phospholipid hydroperoxidation, thereby making them plausible candidates for preventing diseases (i.e. dementia). To evaluate these points, we investigated whether orally administered lutein is distributed to human erythrocytes, and inhibits erythrocyte PLOOH formation. Six healthy subjects took one capsule of food-grade lutein (9.67 mg lutein per capsule) once per d for 4 weeks. Before and during the supplementation period, carotenoids and PLOOH in erythrocytes and plasma were determined by our developed HPLC technique. The administered lutein was incorporated into human erythrocytes, and erythrocyte PLOOH level decreased after the ingestion for 2 and 4 weeks. The antioxidative effect of lutein was confirmed on erythrocyte membranes, but not in plasma. These results suggest that lutein has the potential to act as an important antioxidant molecule in erythrocytes, and it thereby may contribute to the prevention of dementia. Therefore future biological and clinical studies will be required to evaluate the efficacy as well as safety of lutein in models of dementia with a realistic prospect of its use in human therapy.

  5. Effect of glutaraldehyde treatment on enzyme-loaded erythrocytes.

    PubMed

    Deloach, J; Peters, S; Pinkard, O; Glew, R; Ihler, G

    1977-02-28

    In principle, enzyme-loaded erythrocytes can be used as a vehicle for enzyme replacement therapy in lysosomal storage diseases. Glutaraldehyde treatment renders these erythrocytes more resistant to lysis without inactivating the enzymes that have been entrapped inside them. Glutaraldehyde treatment does not prevent ingestion of enzyme-loaded erythrocytes by macrophages in vitro so that these cells can be used to deliver enzymes to lysosomes. In vivo, the glutaraldehyde-treated cells are quickly removed from the circulation by the spleen or liver. The degree of glutaraldehyde treatment allows the erythrocytes to be targeted either to the spleen (low glutaraldehyde concentrations) or to the liver (higher glutaraldehyde concentrations).

  6. Erythrocyte Osmotic Fragility and Excitability Score in Rabbit fed Hibiscus Sabdariffa in Graded Level.

    PubMed

    Adenkola, A Y; Oluremi, O I A

    2014-12-29

    This study was conducted for 10 weeks with the aim of investigating the erythrocyte membrane integrity as measured by erythrocyte osmotic fragility and excitability scores of rabbits fed graded level of Hibiscus sabdariffa calyx (HSC). Twenty weaners' rabbit of both sexes were used for the study and were placed on four experimental diets which contain the following percentages of HSC 0 %, 25 %, 50 %, 75 %, as feed additive and were added at 0 g, 62.5 g, 125 g, 187.5 g designated as T1, T2, T3 and T4 experimental diets. Excitability scores were measured weekly as described by Voisnet et al. (1997). At the end of the experiment, the rabbits were slaughtered by severing the jugular vein. A Blood sample (2 ml) was collected from each rabbit into sampled bottles, containing the Na EDTA as anticoagulant for hematological analysis. Packed cell volume (PCV) Haemoglobin concentration (Hb), Total red blood cell (RBC) count, Total leukocyte count as well as differential leukocyte was determined using standard method. The percentage haemolysis recorded at 0.3 % to 0.8 % was significantly (P < 0.05) higher in rabbits in T1 compared to the remaining 3 diets. The result of excitability score shows that rabbit on diet 1 and 2 had a lower value which was significantly (P < 0.05) lower than rabbits on diets 3 and 4 with a value of 65.5 ± 5.0 and 70.00 ± 5.50 % respectively. In conclusion this study demonstrated for the first time that chronic administration of HSC improves haematological parameters, brain mood and function as well as maintaining erythrocyte membrane integrity.

  7. Identification of the erythrocyte binding domains of Plasmodium vivax and Plasmodium knowlesi proteins involved in erythrocyte invasion

    PubMed Central

    1994-01-01

    Plasmodium vivax and the related monkey malaria, P. knowlesi, require interaction with the Duffy blood group antigen, a receptor for a family of chemokines that includes interleukin 8, to invade human erythrocytes. One P. vivax and three P. knowlesi proteins that serve as erythrocyte binding ligands in such interactions share sequence homology. Expression of different regions of the P. vivax protein in COS7 cells identified a cysteine-rich domain that bound Duffy blood group-positive but not Duffy blood group-negative human erythrocytes. The homologous domain of the P. knowlesi proteins also bound erythrocytes, but had different specificities. The P. vivax and P. knowlesi binding domains lie in one of two regions of homology with the P. falciparum sialic acid binding protein, another erythrocyte binding ligand, indicating conservation of the domain for erythrocyte binding in evolutionarily distant malaria species. The binding domains of these malaria ligands represent potential vaccine candidates and targets for receptor-blockade therapy. PMID:8046329

  8. Effect of propolis on erythrocyte rheology in experimental mercury intoxication in rats.

    PubMed

    Ercis, K; Aydoğan, S; Atayoğlu, A T; Silici, S

    2015-08-01

    In the present study, changes in erythrocyte rheology in association with mercury toxicity and the role of propolis were analyzed in rats. Forty male Wistar Albino rats that were 4-5 months old were used in the study. The control group was administered normal saline intraperitoneal (ip) injections; the mercury chloride group was administered HgCl2 (4 mg/kg, ip); the propolis group was administered propolis (200 mg/kg, by gavage); and the HgCl2+ propolis group was administered HgCl2 (4 mg/kg, ip) + propolis (200 mg/kg, by gavage) for 3 days. The following parameters were analyzed: hematological parameters, plasma potassium (K) levels, methemoglobin, 2,3-DPG, erythrocyte deformability, and hemolysis as a percentage. The results revealed that leukocyte count significantly increased, and a significant decline occurred in the platelet count (p < 0.01). Serum K(+), MetHb, 2, 3-DPG, and hemolysis percentage significantly increased in the rats exposed to mercury (p < 0.01). However, the values of rats administered only with propolis were close to the values of the control group and the changes were avoided by the administration of propolis as protection in the rats exposed to mercury chloride.

  9. The Watt Count System

    NASA Technical Reports Server (NTRS)

    1981-01-01

    The Watt Count System combines aerospace and computer technology, an engineering systems design approach and the environmental control expertise of a group of engineers who worked on Apollo. First step of the system is a computerized energy consumption analysis based on plans for a new home. Company calculates heating and cooling loads and customizes the most energy efficient system for the particular structure analyzed. A quality control engineer assures that insulation and ducting are installed to specifications. Analysis and design techniques are so accurate that company is able to give homeowner a two-year guarantee that the home's energy consumption will not exceed the kilowatt hour level determined by the computer in step one. In an annual energy performance comparison, a Watt Count home used 45 percent less energy than a conventionally equipped home.

  10. Investigation of High-Speed Erythrocyte Flow and Erythrocyte-Wall Impact in a Lab-on-a-Chip.

    PubMed

    Li, Ping; Zheng, Lu; Zhang, Di; Xie, Yonghui; Feng, Yi; Xie, Gongnan

    2016-05-26

    To better understand erythrocyte high-speed motion, collision characteristics, and collision-induced hemolysis probability in rotary blood pumps, a visual experimental investigation of high-speed erythrocyte flow and erythrocyte-wall collision in a lab-on-a-chip was performed. The erythrocyte suspension was driven by a microsyringe pump connected to the microchip, and the erythrocyte flow and erythrocyte-wall impact process were observed and imaged by an optical microscope and a high-speed camera. Two types of microchips with different impact surfaces (flat and curved) were employed. The motion and deformation features before and after collision were studied in detail. The results show that erythrocytes not only move along the flow direction in the flow plane but also rotate and roll in three-dimensional space. Erythrocytes keep discoid shape during the movement in the straight channel, but their deformations during collision are mainly classified into two types: erythrocyte structure is still stable and the erythrocyte performance can be ensured to a certain extent in the TypeA deformation, while the TypeB deformation makes the membrane more likely to fracture on the stretched side, increasing the probability of hemolysis. Furthermore, the movements and deformations of the erythrocytes after collision are analyzed and classified into two types: bouncing and slipping. Moreover, a simulation method for the flow in microchip was performed and validated through a comparison of the streamlines and experimental erythrocytes tracks, which can be further employed to predict the high-speed blood flow, associated with collision process in mechanical blood pump.

  11. Diminished spectrin extraction from ATP-depleted human erythrocytes. Evidence relating spectrin to changes in erythrocyte shape and deformability.

    PubMed

    Lux, S E; John, K M; Ukena, T E

    1978-03-01

    We measured spectrin "extractability" in erythrocytes which were metabolically depleted by incubation at 37 degrees C in plasma or glucose-free buffers. Membranes were extracted with 1 mM EDTA (pH 8, 40 h, 4 degrees C) and analyzed by polyacrylamide gel electrophoresis in sodium dodecyl sulfate. This procedure solubilized 85--90% of the spectrin, actin, and residual hemoglobin from ghosts of fresh erythrocytes. In incubated erythrocytes, inextractable spectrin rapidly accumulated when ATP concentrations fell below 0--15% of normal. In severely depleted cells, 60--90% of the total ghost spectrin became inextractable. Inextractability was not abolished by physically disrupting the ghost before extraction, but was reversed when erythrocyte ATP was replenished with adenosine. The accumulation of inextractable spectrin correlated temporally with the increase in apparent membrane deformability and the increases in erythrocyte vicosity, calcium content, sodium gain, and potassium loss characteristic of ATP-depleted erythrocytes. No change in integral membrane protein topography (assessed by the distribution of intramembranous particles and concanavalin A surface-binding sites) was detected in depleted cells. Analogous changes were observed in erythrocytes exposed to extremes of pH and temperature. When the pH in the erythrocyte interior fell below 5.5, a pH where spectrin was aggregated and isoelectrically precipitated, erythrocyte and ghost viscosity increased coincident with a marked decrease in spectrin extractability. Similarly above 49 degrees C, a temperature where spectrin was denatured and precipitated, erythrocyte viscosity rose as inextractable spectrin accumulated. These observations provide direct evidence of a change in the physical state of spectrin associated with a change in erythrocyte shape and deformability. They support the concept that erythrocyte shape and deformability are largely determined by the shape and deformability of the spectrin

  12. Diminished spectrin extraction from ATP-depleted human erythrocytes. Evidence relating spectrin to changes in erythrocyte shape and deformability.

    PubMed Central

    Lux, S E; John, K M; Ukena, T E

    1978-01-01

    We measured spectrin "extractability" in erythrocytes which were metabolically depleted by incubation at 37 degrees C in plasma or glucose-free buffers. Membranes were extracted with 1 mM EDTA (pH 8, 40 h, 4 degrees C) and analyzed by polyacrylamide gel electrophoresis in sodium dodecyl sulfate. This procedure solubilized 85--90% of the spectrin, actin, and residual hemoglobin from ghosts of fresh erythrocytes. In incubated erythrocytes, inextractable spectrin rapidly accumulated when ATP concentrations fell below 0--15% of normal. In severely depleted cells, 60--90% of the total ghost spectrin became inextractable. Inextractability was not abolished by physically disrupting the ghost before extraction, but was reversed when erythrocyte ATP was replenished with adenosine. The accumulation of inextractable spectrin correlated temporally with the increase in apparent membrane deformability and the increases in erythrocyte vicosity, calcium content, sodium gain, and potassium loss characteristic of ATP-depleted erythrocytes. No change in integral membrane protein topography (assessed by the distribution of intramembranous particles and concanavalin A surface-binding sites) was detected in depleted cells. Analogous changes were observed in erythrocytes exposed to extremes of pH and temperature. When the pH in the erythrocyte interior fell below 5.5, a pH where spectrin was aggregated and isoelectrically precipitated, erythrocyte and ghost viscosity increased coincident with a marked decrease in spectrin extractability. Similarly above 49 degrees C, a temperature where spectrin was denatured and precipitated, erythrocyte viscosity rose as inextractable spectrin accumulated. These observations provide direct evidence of a change in the physical state of spectrin associated with a change in erythrocyte shape and deformability. They support the concept that erythrocyte shape and deformability are largely determined by the shape and deformability of the spectrin

  13. Preferential Elimination of Older Erythrocytes in Circulation and Depressed Bone Marrow Erythropoietic Activity Contribute to Cadmium Induced Anemia in Mice

    PubMed Central

    Chatterjee, Sreoshi; Saxena, Rajiv K.

    2015-01-01

    Feeding cadmium chloride (50 or 1000 ppm CdCl2 in drinking water, ad libitum) to C57BL/6 mice resulted in a significant and sustained fall in blood erythrocyte count and hemoglobin levels that started 4 and 3 weeks after the start of 50 and 1000 ppm cadmium doses respectively. A transient yet significant reticulocytosis occurred during the first 4 weeks of cadmium treatment. Using the recently developed double in vivo biotinylation (DIB) technique, turnover of erythrocyte cohorts of different age groups was simultaneously monitored in control and cadmium treated mice. A significant accumulation of younger erythrocytes and a concomitant decline in the relative proportions of older erythrocytes in circulation was observed in both 50 and 1000 ppm cadmium groups indicating that older erythrocytes were preferentially eliminated in cadmium induced anemia. A significant increase in the erythropoietin levels in plasma was seen in mice exposed to 1000 ppm cadmium. Levels of inflammatory cytokines (IL1A, IL6, TNFα, IFNγ) were however not significantly altered in cadmium treated mice. A significant increase in cellular levels of reactive oxygen species (ROS) was observed in older erythrocytes in circulation but not in younger erythrocytes. Erythropoietic activity in the bone marrows and spleens of cadmium treated mice was examined by monitoring the relative proportion of cells belonging to the erythroid line of differentiation in these organs. Erythroid cells in bone marrow declined markedly (about 30%) in mice in the 1000 ppm cadmium group but the decline was not significant in the 50 ppm cadmium group. Cells representing various stages of erythroid differentiation in bone marrow and spleen were enumerated flow cytometrically by double staining with anti-Ter119 and anti-transferrin receptor (CD71) monoclonal antibodies. Decline of erythroid cells was essentially confined to pro-erythroblast and erythroblast-A, along with a concurrent increase in the splenic erythroid

  14. Preferential Elimination of Older Erythrocytes in Circulation and Depressed Bone Marrow Erythropoietic Activity Contribute to Cadmium Induced Anemia in Mice.

    PubMed

    Chatterjee, Sreoshi; Saxena, Rajiv K

    2015-01-01

    Feeding cadmium chloride (50 or 1000 ppm CdCl2 in drinking water, ad libitum) to C57BL/6 mice resulted in a significant and sustained fall in blood erythrocyte count and hemoglobin levels that started 4 and 3 weeks after the start of 50 and 1000 ppm cadmium doses respectively. A transient yet significant reticulocytosis occurred during the first 4 weeks of cadmium treatment. Using the recently developed double in vivo biotinylation (DIB) technique, turnover of erythrocyte cohorts of different age groups was simultaneously monitored in control and cadmium treated mice. A significant accumulation of younger erythrocytes and a concomitant decline in the relative proportions of older erythrocytes in circulation was observed in both 50 and 1000 ppm cadmium groups indicating that older erythrocytes were preferentially eliminated in cadmium induced anemia. A significant increase in the erythropoietin levels in plasma was seen in mice exposed to 1000 ppm cadmium. Levels of inflammatory cytokines (IL1A, IL6, TNFα, IFNγ) were however not significantly altered in cadmium treated mice. A significant increase in cellular levels of reactive oxygen species (ROS) was observed in older erythrocytes in circulation but not in younger erythrocytes. Erythropoietic activity in the bone marrows and spleens of cadmium treated mice was examined by monitoring the relative proportion of cells belonging to the erythroid line of differentiation in these organs. Erythroid cells in bone marrow declined markedly (about 30%) in mice in the 1000 ppm cadmium group but the decline was not significant in the 50 ppm cadmium group. Cells representing various stages of erythroid differentiation in bone marrow and spleen were enumerated flow cytometrically by double staining with anti-Ter119 and anti-transferrin receptor (CD71) monoclonal antibodies. Decline of erythroid cells was essentially confined to pro-erythroblast and erythroblast-A, along with a concurrent increase in the splenic erythroid

  15. Erythrocyte membrane tropomyosin. Purification and properties.

    PubMed

    Fowler, V M; Bennett, V

    1984-05-10

    Two polypeptides of Mr approximately 29,000 and 27,000 have been identified in human erythrocyte membranes that cross-react specifically with affinity purified antibodies to chicken gizzard tropomyosin. The cross-reacting polypeptides are quantitatively retained on the membrane after cell lysis if millimolar concentrations of magnesium are included in the lysis and wash buffers, indicating that they are membrane-bound proteins under physiological conditions. Milligram quantities of these immunoreactive polypeptides have been purified to greater than 95% purity from a low salt extract of membranes by DEAE-chromatography, precipitation at pH 4.4, and heating to 85 degrees C to denature contaminants. Physical similarities of the erythrocyte protein to other tropomyosins include (a) amino acid composition (b) anomalous migration of the Mr approximately 29,000 and 27,000 polypeptides on sodium dodecyl sulfate-gels in the presence of 6 M urea to apparent Mr approximately 43,000 and 38,000, respectively (c) arrangement of chains as dimers of Mr approximately 60,000 based on cross-linking studies and calculation of molecular weight from hydrodynamic values (Rs = 5.9 nm, sedimentation coefficient = 2.5 S; partial specific volume = 0.72 cm3/g) and (d) highly asymmetric shape, based on a frictional ratio of 2.07. Binding of erythrocyte tropomyosin to muscle F-actin saturates at one tropomyosin molecule (Mr approximately 60,000) to 6-7 actin monomers and is highly cooperative with a Hill coefficient of about 2.8, similar to muscle tropomyosins. Binding also exhibits a high degree of cooperativity as a function of the magnesium concentration with a transition between no binding and complete binding between 1 and 2 mM MgCl2. Increasing the magnesium concentration from 2 to 10 mM increases the apparent affinity of tropomyosin for actin from approximately 2.6 X 10(6) M-1 to approximately 2.7 X 10(7) M-1 without effect on the Hill coefficient. The tropomyosin polypeptides comprise

  16. Plasmodium falciparum AMA-1 erythrocyte binding peptides implicate AMA-1 as erythrocyte binding protein.

    PubMed

    Urquiza, M; Suarez, J E; Cardenas, C; Lopez, R; Puentes, A; Chavez, F; Calvo, J C; Patarroyo, M E

    2000-10-15

    The role of AMA-1 during merozoite invasion has not yet been determined. However, reported experimental evidence suggests that this protein can be used, in particular as erythrocyte-binding protein, since, Fab fragments against this protein are able to block merozoite invasion. Using a previously described methodology, eight peptides with high binding activity to human erythrocyte, scattered along the different domains and having around 130 nM affinity constants, were identified in the Plasmodium falciparum AMA-1 protein. Their binding activity was sialic acid independent. Some of these peptides showed homology with the erythrocyte binding domains of one of the apical organelle protein family, MAEBL, identified in rodent malarial parasites. One of these peptides shares amino acid sequence with a previously reported B-cell epitope which induces antibodies to block parasite growth. The critical residues were identified for erythrocyte binding conserved peptides 4313 (DAEVAGTQYRLPSGKCPVFG), 4321 (VVDNWEKVCPRKNLQNAKFG), 4325 (MIKSAFLPTGAFKADRYKSH) and 4337 (WGEEKRASHTTPVLMEKPYY). All conserved peptides were able to block merozoite invasion of new RBC and development, suggesting that these peptides are involved in P. falciparum invasion.

  17. Erythrocytic vacuolar rafts induced by malaria parasites.

    PubMed

    Haldar, K; Samuel, B U; Mohandas, N; Harrison, T; Hiller, N L

    2001-03-01

    Studies in the past year displaced long-standing dogmas and provided many new molecular insights into how proteins and solutes move between the erythrocyte plasma membrane and the malarial vacuole. Highlights include a demonstration that (1) detergent-resistant membrane (DRM) rafts exist in the red cell membrane and their resident proteins are detected as rafts in the plasmodial vacuole, (2) a voltage-gated channel in the infected red cell membrane mediates uptake of extracellular nutrient solutes, and (3) intraerythrocytic membranes transport a parasite-encoded adherence antigen to the red cell surface.

  18. Alterations of erythrocyte membrane organization in alcoholics.

    PubMed

    Beaugé, F; Stibler, H; Borg, S

    1987-01-01

    Studies of fluorescence polarization of DPH have shown that erythrocyte membrane "fluidity" and fluidization by ethanol are significantly reduced in alcoholics. By using probes of the polar part of the membrane, ANS and TMA-DPH, in addition to DPH, it was shown in the present study that disturbances also exist in the polar region of the membrane which probably are related to changes in surface glycoconjugates. Furthermore, the acute fluidizing effect of ethanol was correlated with the capacity of the membrane to bind ethanol, which in turn appeared to be linked to the glycans. Chronic ethanol abuse thus causes complex disturbances of membrane organization at different levels of the membrane.

  19. The influence of various hematology analyzers on component platelet counts.

    PubMed

    Moroff, Gary; Sowemimo-Coker, Samuel O; Finch, Stephen; Murphy, Scott; Brandwein, Harvey; Whitbread, John; Wenz, Barry

    2005-04-01

    Hematology analyzers designed to count platelets in samples of whole blood are used to enumerate the total number of platelets in components prepared for transfusion. This report addresses the issue of variability in platelet counts obtained with different models of hematology analyzers. The influence of a common calibration procedure, involving one level of porcine platelets, on the extent of variability was also evaluated. Identical sets of samples of simulated and apheresis-derived human platelets were counted by multiple laboratories in 3 separate studies. In the first 2 exercises, 7 samples of both porcine platelets and modified goat erythrocytes with targeted platelets counts from 0.2 to 4.0 x 10(12)/L were counted without prior dilution. In both exercises, the samples were counted multiple times after routine calibration using instructions provided by the manufacturers of the various hematology analyzers used. In the second exercise, the samples were recounted after the hematology analyzers were recalibrated with a common calibrant consisting of porcine platelets at a targeted concentration of 0.5 x 10(12)/L. In the first and second exercises, 20 and 18 hematology analyzers were used, respectively. In the third exercise, 6 samples prepared from a single unit of apheresis platelets with targeted counts from 0.2 to 1.64 x 10(12)/L were shipped by an overnight courier and counted in triplicate on the day of arrival. Eleven hematology analyzers were used. The influence of recalibration was evaluated statistically by using the 95% prediction interval for the mean of a future set of observations. The platelet counts measured with a specific type of hematology analyzer provided the data to calculate the 95% prediction interval. With routine calibration, a wide variability in platelet counts was observed with all levels of both simulated and apheresis-derived human platelets. For example, with porcine platelets at a targeted level of 0.4 x 10 (12)/L, the platelet

  20. Erythrocyte membrane skeleton inhibits nanoparticle endocytosis

    NASA Astrophysics Data System (ADS)

    Gao, Xinli; Yue, Tongtao; Tian, Falin; Liu, Zhiping; Zhang, Xianren

    2017-06-01

    Red blood cells (RBCs), also called erythrocytes, have been experimentally proposed in recent decades as the biological drug delivery systems through entrapping certain drugs by endocytosis. However, the internalization pathway of endocytosis seems to conflict with the robust mechanical properties of RBCs that is induced by the spectrin-actin network of erythrocyte membrane skeleton. In this work, we employed a minimum realistic model and the dissipative particle dynamics method to investigate the influence of the spectrin-actin membrane skeleton on the internalization of nanoparticles (NPs). Our simulations show that the existence of skeleton meshwork indeed induces an inhibiting effect that effectively prevents NPs from internalization. The inhibiting effect is found to depend on the membrane-NP attraction, skeleton tension and relative size of the NP to the membrane skeleton mesh. However, our simulations also demonstrate that there are two possibilities for successful internalization of NPs in the presence of the membrane skeleton. The first case is for NPs that has a much smaller size than the dimension of skeleton meshes, and the other is that the skeleton tension is rather weak so that the formed vesicle can still move inward for NP internalization.

  1. Binding characteristics of swine erythrocyte insulin receptors

    SciTech Connect

    Dieberg, G.; Bryan, G.S.; Sartin, J.L.; Williams, J.C.; Prince, T.J.; Kemppainen, R.J.

    1985-09-01

    Crossbred gilts had 8.8 +/- 1.1% maximum binding of ( SVI)insulin to insulin receptors on erythrocytes. The number of insulin-binding sites per cell was 137 +/- 19, with a binding affinity ranging from 7.4 X 10(7)M-1 to 11.2 X 10(7)M-1 and mean of 8.8 X 10(7)M-1. Pregnant sows had a significant increase in maximum binding due to an increase in number of receptor sites per cell. Lactating sows fed a high-fiber diet and a low-fiber diet did not develop a significant difference in maximum binding of insulin. Sows fed the low-fiber diet had a significantly higher number of binding sites and a significantly lower binding affinity than did sows fed a high-fiber diet. Receptor-binding affinity was lower in the low-fiber diet group than in cycling gilts, whereas data from sows fed the high-fiber diet did not differ from data for cycling gilts. Data from this study indicated that insulin receptors of swine erythrocytes have binding characteristics similar to those in other species. Pregnancy and diet will alter insulin receptor binding in swine.

  2. Mycoplasma gallisepticum invades chicken erythrocytes during infection.

    PubMed

    Vogl, Gunther; Plaickner, Astrid; Szathmary, Susan; Stipkovits, László; Rosengarten, Renate; Szostak, Michael P

    2008-01-01

    Recently, it was demonstrated using in vitro assays that the avian pathogen Mycoplasma gallisepticum is able to invade nonphagocytic cells. It was also shown that this mycoplasma can survive and multiply intracellularly for at least 48 h and that this cell invasion capacity contributes to the systemic spread of M. gallisepticum from the respiratory tract to the inner organs. Using the gentamicin invasion assay and a differential immunofluorescence technique combined with confocal laser scanning microscopy, we were able to demonstrate in in vitro experiments that M. gallisepticum is also capable of invading sheep and chicken erythrocytes. The frequencies of invasion of three well-defined M. gallisepticum strains were examined over a period of 24 h, and a significant increase in invasiveness occurred after 8 h of infection. In addition, blood samples derived from chickens experimentally infected via the aerosol route with the virulent strain M. gallisepticum R(low) were analyzed. Surprisingly, M. gallisepticum R(low) was detected in the bloodstream of infected chickens by nested PCR, as well as by differential immunofluorescence and interference contrast microscopy that showed that mycoplasmas were not only on the surface but also inside chicken erythrocytes. This finding provides novel insight into the pathomechanism of M. gallisepticum and may have implications for the development of preventive strategies.

  3. Efflux and influx of erythrocyte water.

    PubMed

    OLMSTEAD, E G

    1960-11-01

    Rabbit erythrocytes were washed in buffered NaCl solutions isotonic with rabbit serum (Delta(t) -0.558 degrees C.) and suspended in buffered NaCl solutions of tonicity equidistant from intracellular tonicity (Delta(t) = -0.558 degrees C. +/- 0.112 degrees C.) of varying pH and incubated at varying temperatures. After incubation, the freezing point depression (Delta(t)) was measured on the supernatant. Change in the Delta(t) measured change in the water content of the extracellular solutions-water being withdrawn by erythrocytes (W(I)) from the hypotonic solutions and added (W(E)) to the hypertonic solutions. W(E) was always less than W(I) and was inversely proportional to the pH in the range 6.5-8.0. W(E) was significantly increased by lowering the temperature of the cell suspension to 4 degrees C. W(I) was increased by raising or lowering the pH or raising the temperature of the cell suspension. W(E) x W(I) not equal k. W(E) and W(I) were affected differently by changes in pH and temperature. It was concluded that W(E) and W(E) were probably under different physicochemical control.

  4. Erythrocyte and platelet proteomics in hematological disorders.

    PubMed

    Chakrabarti, Abhijit; Halder, Suchismita; Karmakar, Shilpita

    2016-04-01

    Erythrocytes undergo ineffective erythropoesis, hemolysis, and premature eryptosis in sickle cell disease and thalassemia. Abnormal hemoglobin variants associated with hemoglobinopathy lead to vesiculation, membrane instability, and loss of membrane asymmetry with exposal of phosphatidylserine. This potentiates thrombin generation resulting in activation of the coagulation cascade responsible for subclinical phenotypes. Platelet activation also results in the release of microparticles, which express and transfer functional receptors from platelet membrane, playing key roles in vascular reactivity and activation of intracellular signaling pathways. Over the last decade, proteomics had proven to be an important field of research in studies of blood and blood diseases. Blood cells and its fluidic components have been proven to be easy systems for studying differential expressions of proteins in hematological diseases encompassing hemoglobinopathies, different types of anemias, myeloproliferative disorders, and coagulopathies. Proteomic studies of erythrocytes and platelets reported from several groups have highlighted various factors that intersect the signaling networks in these anucleate systems. In this review, we have elaborated on the current scenario of anucleate blood cell proteomes in normal and diseased individuals and the cross-talk between the two major constituent cell types of circulating blood.

  5. Comparative erythrocyte metabolism in marsupials and monotremes.

    PubMed

    Parkinson, A L; Whittington, A T; Spencer, P B; Grigg, G; Hinds, L; Gallagher, C; Kuchel, P; Agar, N S

    1995-03-01

    Concentrations of ATP and DPG, activities of 10 enzymes and the glycolytic rates were measured in the erythrocytes of 11 species of marsupials and two species of monotremes. Mean DPG concentrations were greater in the erythrocytes of marsupials than those of eutherian mammals. The opposite is true of ATP. Significant findings from the results of enzyme activities were: high activity of hexokinase (7.39 +/- 0.82 EU/g Hb) in the short-beaked echidna, pyruvate kinase (37.49 +/- 1.0 EU/g) Hb in bridled nailtail wallaby and glucose-6-phosphate dehydrogenase (G6PD; 41.66 +/- 1.24 EU/g Hb) in black-striped wallaby. About 6- to 7-fold difference in the activity of G6PD levels between the two species of wombats was confirmed. Glucose phosphate isomerase activity was also shown to be twice as high in the red cells of the common wombat compared with those of the southern hairy nosed wombat. There were wide variations in the glycolytic rate among the species examined.

  6. Phlorhizin protects against erythrocyte cell membrane scrambling.

    PubMed

    Gatidis, Sergios; Meier, Anja; Jilani, Kashif; Lang, Elisabeth; Zelenak, Christine; Qadri, Syed M; Lang, Florian

    2011-08-10

    Phlorhizin interferes with glucose transport. Glucose depletion triggers suicidal erythrocyte death or eryptosis, characterized by cell shrinkage and cell membrane scrambling. Eryptosis is further triggered by oxidative stress. The present study explored whether phlorhizin influences eryptosis following glucose depletion or oxidative stress. Cell membrane scrambling was estimated from annexin binding, cell volume from forward scatter (FSC), and cytosolic Ca(2+) concentration from Fluo-3 fluorescence. Phlorhizin (10-100 μM) added alone did not modify scrambling, FSC, or Fluo-3 fluorescence. Glucose depletion (48 h) significantly increased Fluo-3 fluorescence, decreased FSC, and increased annexin binding, effects in part significantly blunted by phlorhizin (annexin binding ≥ 10 μM, FSC ≥ 50 μM). Oxidative stress (30 min 0.3 mM tert-butylhydroperoxide) again significantly increased Fluo-3 fluorescence and triggered annexin binding, effects again in part significantly blunted by phlorhizin (Fluo-3 fluorescence ≥ 50 μM, annexin-binding ≥ 10 μM). Phlorhizin did not blunt the cell shrinkage induced by oxidative stress. The present observations disclose a novel effect of phlorhizin, that is, an influence on suicidal erythrocyte death following energy depletion and oxidative stress.

  7. Inhibition of suicidal erythrocyte death by probucol.

    PubMed

    Shaik, Nazneen; Lupescu, Adrian; Lang, Florian

    2013-02-01

    Probucol, an antioxidant and anti-inflammatory agent counteracting atherosclerosis and restenosis, is partially effective by influencing suicidal cell death or apoptosis. In analogy to apoptosis of nucleated cells, suicidal death of erythrocytes or eryptosis is characterized by cell shrinkage and cell membrane scrambling with phosphatidylserine exposure at the erythrocyte surface. Eryptosis is stimulated by increase in cytosolic Ca(2+) activity, for example, after energy depletion or oxidative stress. The present study explored whether probucol influences eryptosis. Phosphatidylserine exposure was estimated from annexin-V-binding, cell volume from forward scatter (FSC), and cytosolic Ca(2+) concentration from fluo-3 fluorescence in flow cytometry. As a result, energy depletion (48-hour glucose removal) increased annexin-V-binding, decreased FSC, and increased fluo-3 fluorescence. Probucol (≤30 μM) did not significantly modify annexin-V-binding, FSC, or fluo-3 fluorescence in the presence of glucose but (at ≥5 μM) blunted the effect of glucose depletion on annexin-V-binding. Probucol (≥20 μM) only slightly blunted the effects of glucose depletion on FSC and fluo-3 fluorescence. Ca(2+) ionophore ionomycin (1 μM) and oxidative stress (30-minute exposure to 0.3 mM of tert-butylhydroperoxide) increased annexin-V-binding, effects again blunted by 30 μM of probucol. In conclusion, probucol blunts cell membrane scrambling after energy depletion and oxidative stress, effects primarily because of interference with the scrambling effects of increased cytosolic Ca(2+) concentration.

  8. Dielectric properties and ion mobility in erythrocytes.

    PubMed

    Pauly, H; Schwan, H P

    1966-09-01

    The impedance of erythrocytes of man, cattle, sheep, dog, cat, rabbit, and chicken was measured in the range from 0.5 to 250 Mc. The dielectric constant of the red cell interior is 50 at 250 Mc, varies but little with species, and can readily be accounted for by the cells' hemoglobin content. The electrical conductivity of the red cell interior was determined between 70 and 100 Mc. The values differ from species to species within the rather limited range from 4.4 to 5.3 mmho/cm. Removal of the cell membranes does not affect the conductivity. Hence, the cell interior behaves, from an electrical point of view, like a highly concentrated hemoglobin solution. A theoretical value for the electrical conductivity of erythrocyte interiors, which is calculated on the basis of the salt content of the cell, ion mobility, and the volume concentration of the hemoglobin, is roughly twice as large as the measured value. This discrepancy is typical not only of the red blood cell. Pertinent measurements show that it is probably caused by hydrodynamic and possibly by electrostatic effects also, which lower the mobility of the ions. From the lower electrical mobility it appears that a lowered diffusion constant of the electrolytes and nonelectrolytes within the cell is indicated.

  9. 21 CFR 864.6700 - Erythrocyte sedimentation rate test.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 21 Food and Drugs 8 2012-04-01 2012-04-01 false Erythrocyte sedimentation rate test. 864.6700 Section 864.6700 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES HEMATOLOGY AND PATHOLOGY DEVICES Manual Hematology Devices § 864.6700 Erythrocyte...

  10. 21 CFR 864.6700 - Erythrocyte sedimentation rate test.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 21 Food and Drugs 8 2013-04-01 2013-04-01 false Erythrocyte sedimentation rate test. 864.6700 Section 864.6700 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES HEMATOLOGY AND PATHOLOGY DEVICES Manual Hematology Devices § 864.6700 Erythrocyte...

  11. 21 CFR 864.6700 - Erythrocyte sedimentation rate test.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 21 Food and Drugs 8 2011-04-01 2011-04-01 false Erythrocyte sedimentation rate test. 864.6700 Section 864.6700 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES HEMATOLOGY AND PATHOLOGY DEVICES Manual Hematology Devices § 864.6700 Erythrocyte...

  12. 21 CFR 864.6700 - Erythrocyte sedimentation rate test.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 21 Food and Drugs 8 2014-04-01 2014-04-01 false Erythrocyte sedimentation rate test. 864.6700 Section 864.6700 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES HEMATOLOGY AND PATHOLOGY DEVICES Manual Hematology Devices § 864.6700 Erythrocyte...

  13. Effect of laser irradiation of donor blood on erythrocyte shape.

    PubMed

    Baibekov, I M; Ibragimov, A F; Baibekov, A I

    2012-04-01

    Changes in erythrocyte shape in donor blood during storage and after irradiation with He-Ne laser and infrared laser were studied by scanning electron microscopy, thick drop express-method, and morphometry. It was found that laser irradiation delayed the appearance of erythrocytes of pathological shapes (echinocytes, stomatocytes, etc.) in the blood; He-Ne laser produced a more pronounced effect.

  14. High Red Blood Cell Count

    MedlinePlus

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

  15. Changes in the complete blood count and blood rheology in patients after myocardial infarction participating in the rehabilitation programme.

    PubMed

    Pabisiak, A; Bromboszcz, J; Kmiec, S; Dendura, M; Dabrowski, Z; Smolenski, O

    2015-01-01

    The aim of cardiovascular disease treatment is to reduce the risk of thrombogenesis and improve tissue perfusion, depending inter alia on the rheological properties of the blood. The reduction in blood viscosity and erythrocyte aggregation, as well as increase of erythrocyte deformability was observed under the influence of physical training. To compare the blood count and rheological properties of blood samples before and after outpatient cardiac rehabilitation programme. 35 men (average age: 57.2 ± 5.42), who after suffering myocardial infarction treated with percutaneous coronary intervention (PCI), took part in 24 physical training sessions of moderate intensity (40-60% of heart rate reserve). The standard ergometer submaximal (up to 85% of predicted HRmax) exercise test and echocardiography was performed before and after training. Blood count, fibrinogen concentration as well as aggregation and elongation properties of erythrocytes were analyzed too. Patients significantly increased exercise capacity (p <  0.00001) and ejection fraction (p <  0.00001) after completion of the training cycle. There was noted a reduction in aggregation index AI (p <  0.01), an increase in the number of erythrocytes RBC (p <  0.05), while reducing their volume MCV (p <  0.05) and a decrease in hemoglobin content MCH (p <  0.05). The number of leukocytes WBC (p <  0.01) was decreased too. Beneficial changes in blood rheology and blood count have been found in patients after myocardial infarction who took part in the physical training sessions of moderate intensity.

  16. A role of phosphatidylserine externalization in clearance of erythrocytes exposed to stress but not in eliminating aging populations of erythrocyte in mice.

    PubMed

    Khandelwal, Sanjay; Saxena, Rajiv K

    2008-08-01

    Age dependent changes in phosphatidylserine (PS) externalization were studied in mouse erythrocytes of different age groups (range 1-55 days) by using a newly developed double in vivo biotinylation (DIB) technique. Around 3-4% of the erythrocytes freshly released in the circulation were PS(+) but this proportion fell rapidly to 1% or less and did not increase at later time points. Blocking erythrocyte clearance from the circulation by in vivo depletion of macrophages (by treatment with clodronate loaded liposomes) for up to 7 days did not result in accumulation of PS(+) erythrocytes in the circulation indicating that the low percentage of PS(+) cells within old erythrocytes (age >40 days) was not related to the clearance of PS(+) erythrocytes by macrophages. In vitro treatment with stress inducing agents like deoxyglucose or Ca(++)/calcium ionophore resulted in a marked induction of PS externalization in mouse erythrocytes and this effect was most prominent in the youngest erythrocyte population (age <10 days). Kinetics of clearance of different age groups of stress exposed erythrocytes after intravenous infusion into recipient mice indicated that the young erythrocytes were cleared at fastest rate from the circulation as compared to erythrocytes of older age groups. Within young erythrocytes exposed to stress, PS(+) erythrocytes were preferentially cleared. Taken together our results suggest that PS externalization is unlikely to have a role in the removal of old erythrocytes from blood circulation but may have a role in the clearance of stressed and damaged young erythrocytes in blood circulation.

  17. Effect of phytic acid on suicidal erythrocyte death.

    PubMed

    Eberhard, Matthias; Föller, Michael; Lang, Florian

    2010-02-10

    Phytic acid, an anticarcinogenic food component, stimulates apoptosis of tumor cells. Similar to apoptosis, human erythrocytes may undergo suicidal death or eryptosis, characterized by cell membrane scrambling and cell shrinkage. Triggers of eryptosis include energy depletion. Phytate intake could cause anemia, an effect attributed to iron complexation. The present experiments explored whether phytic acid influences eryptosis. Supernatant hemoglobin concentration was determined to reveal hemolysis, annexin V-binding in FACS analysis was utilized to identify erythrocytes with scrambled cell membrane, forward scatter in FACS analysis was taken as a measure of cell volume, and a luciferin-luciferase assay was employed to determine erythrocyte ATP content. As a result, phytic acid (>or=1 mM) did not lead to significant hemolysis, but significantly increased the percentage of annexin V-binding erythrocytes, significantly decreased forward scatter, and significantly decreased cellular ATP content. In conclusion, phytic acid stimulates suicidal human erythrocyte death, an effect paralleling its proapoptotic effect on nucleated cells.

  18. Encapsulation of thiosulfate: cyanide sulfurtransferase by mouse erythrocytes

    SciTech Connect

    Leung, P.; Ray, L.E.; Sander, C.; Way, J.L.; Sylvester, D.M.; Way, J.L.

    1986-03-30

    Murine carrier erythrocytes, prepared by hypotonic dialysis, were employed in the encapsulation of several compounds including (14C)sucrose, (3H)inulin, and bovine thiosulfate:cyanide sulfurtransferase (rhodanese), a mitochondrial enzyme which converts cyanide to thiocyanate. Approximately 30% of the added (14C)sucrose, (3H)inulin, and rhodanese was encapsulated by predialyzed erythrocytes, and a decrease in the mean corpuscular volume and mean corpuscular hemoglobin was observed. In the encapsulation of rhodanese a recovery of 95% of the erythrocytes was achieved and an 85% equilibrium was established. The addition of potassium cyanide (50 mM) to intact, rhodanese-loaded erythrocytes containing sodium thiosulfate resulted in its metabolism to thiocyanate. These results establish the potential use of erythrocytes as biodegradable drug carrier in drug antagonism.

  19. Atomic force microscopic observation of surface-supported human erythrocytes

    NASA Astrophysics Data System (ADS)

    Ho, Mon-Shu; Kuo, Feng-Jia; Lee, Yu-Siang; Cheng, Chao-Min

    2007-07-01

    The nanomechanical characteristics of the membrane cytoskeleton of human erythrocytes were studied using atomic force microscopy (AFM). The self-assembly, fine structure, cell diameter, thickness, and reticulate cytoskeleton of erythrocytes on the mica surface were investigated. The adhesive forces that correspond to the membrane elasticity of various parts of the erythrocyte membrane surface were measured directly by AFM to be 0.64±0.14nN for cell indentation, 4.2±0.7nN for cell hump, and 11.5nN for side waist, respectively. The deformation of erythrocytes was discussed. Standing waves on the membrane that were set up by increased AFM amplitude were observed. The propagating velocity on the erythrocyte membrane was estimated to be ˜2.02×10-2m/s. Liquid physiological conditions were considered throughout.

  20. Light-induced protoporphyrin release from erythrocytes in erythropoietic protoporphyria

    SciTech Connect

    Sandberg, S.; Brun, A.

    1982-09-01

    The photohemolysis of normal erythrocytes incubated with protoporphyrin is reduced in the presence of albumin. When globin is added to normal erythrocytes loaded with protoporphyrin, protoporphyrin is bound to globin. During irradiation protoporphyrin moves from globin to the erythrocyte membrane and photohemolysis is initiated. Erythrocytes in patients with erythropoietic protoporphyria contain large amounts of protoporphyrin bound to hemoglobin. Upon irradiation of these cells in the absence of albumin, 40% of protoporphyrin and 80% of hemoglobin is released after 240 kJ/m2. The released protoporphyrin is hemoglobin bound. In contrast, when albumin is present only 8% of hemoglobin is released whereas protoporphyrin is released to 76%. The released protoporphyrin is albumin bound. A hypothesis for the release of erythrocyte protoporphyrin in erythropoietic protoporphyria without simultaneous hemolysis is proposed. Upon irradiation protoporphyrin photodamages its binding sites on hemoglobin, moves through the plasma membrane, and is bound to albumin in plasma.

  1. Antioxidant effect of astaxanthin on phospholipid peroxidation in human erythrocytes.

    PubMed

    Nakagawa, Kiyotaka; Kiko, Takehiro; Miyazawa, Taiki; Carpentero Burdeos, Gregor; Kimura, Fumiko; Satoh, Akira; Miyazawa, Teruo

    2011-06-01

    Phospholipid hydroperoxides (PLOOH) accumulate abnormally in the erythrocytes of dementia patients, and dietary xanthophylls (polar carotenoids such as astaxanthin) are hypothesised to prevent the accumulation. In the present study, we conducted a randomised, double-blind, placebo-controlled human trial to assess the efficacy of 12-week astaxanthin supplementation (6 or 12 mg/d) on both astaxanthin and PLOOH levels in the erythrocytes of thirty middle-aged and senior subjects. After 12 weeks of treatment, erythrocyte astaxanthin concentrations were higher in both the 6 and 12 mg astaxanthin groups than in the placebo group. In contrast, erythrocyte PLOOH concentrations were lower in the astaxanthin groups than in the placebo group. In the plasma, somewhat lower PLOOH levels were found after astaxanthin treatment. These results suggest that astaxanthin supplementation results in improved erythrocyte antioxidant status and decreased PLOOH levels, which may contribute to the prevention of dementia.

  2. Ferrokinetic and erythrocyte survival studies in healthy and anemic cats

    SciTech Connect

    Madewell, B.R.; Holmes, P.H.; Onions, D.E.

    1983-03-01

    Erythrocyte survival and ferrokinetic studies were adapted to the cat. For 5 clinically healthy 4- to 9-month-old cats, mean /sup 51/Cr-labeled erythrocyte survival was 144 hours, and mean plasma /sup 59/Fe-labeled transferrin disappearance halftime was 51 minutes. Erythrocyte use of radioiron was rapid and efficient, with 50% to 80% of labeled iron incorporated into the erythron by 100 hours after injection into the cat. Six cats with feline leukemia virus infection were studied. For 2 cats with erythroid aplasia associated with C subgroup of feline leukemia virus, erythrocyte survival times were similar to those determined for the healthy cats, but plasma radioiron disappearance half time and erythrocyte use of radioiron were markedly diminished.

  3. Neural networks counting chimes.

    PubMed Central

    Amit, D J

    1988-01-01

    It is shown that the ideas that led to neural networks capable of recalling associatively and asynchronously temporal sequences of patterns can be extended to produce a neural network that automatically counts the cardinal number in a sequence of identical external stimuli. The network is explicitly constructed, analyzed, and simulated. Such a network may account for the cognitive effect of the automatic counting of chimes to tell the hour. A more general implication is that different electrophysiological responses to identical stimuli, at certain stages of cortical processing, do not necessarily imply synaptic modification, a la Hebb. Such differences may arise from the fact that consecutive identical inputs find the network in different stages of an active temporal sequence of cognitive states. These types of networks are then situated within a program for the study of cognition, which assigns the detection of meaning as the primary role of attractor neural networks rather than computation, in contrast to the parallel distributed processing attitude to the connectionist project. This interpretation is free of homunculus, as well as from the criticism raised against the cognitive model of symbol manipulation. Computation is then identified as the syntax of temporal sequences of quasi-attractors. PMID:3353371

  4. Counting RG flows

    DOE PAGES

    Gukov, Sergei

    2016-01-05

    Here, interpreting renormalization group flows as solitons interpolating between different fixed points, we ask various questions that are normally asked in soliton physics but not in renormalization theory. Can one count RG flows? Are there different "topological sectors" for RG flows? What is the moduli space of an RG flow, and how does it compare to familiar moduli spaces of (supersymmetric) dowain walls? Analyzing these questions in a wide variety of contexts -- from counting RG walls to AdS/CFT correspondence -- will not only provide favorable answers, but will also lead us to a unified general framework that is powerfulmore » enough to account for peculiar RG flows and predict new physical phenomena. Namely, using Bott's version of Morse theory we relate the topology of conformal manifolds to certain properties of RG flows that can be used as precise diagnostics and "topological obstructions" for the strong form of the C-theorem in any dimension. Moreover, this framework suggests a precise mechanism for how the violation of the strong C-theorem happens and predicts "phase transitions" along the RG flow when the topological obstruction is non-trivial. Along the way, we also find new conformal manifolds in well-known 4d CFT's and point out connections with the superconformal index and classifying spaces of global symmetry groups.« less

  5. Counting RG flows

    SciTech Connect

    Gukov, Sergei

    2016-01-05

    Here, interpreting renormalization group flows as solitons interpolating between different fixed points, we ask various questions that are normally asked in soliton physics but not in renormalization theory. Can one count RG flows? Are there different "topological sectors" for RG flows? What is the moduli space of an RG flow, and how does it compare to familiar moduli spaces of (supersymmetric) dowain walls? Analyzing these questions in a wide variety of contexts -- from counting RG walls to AdS/CFT correspondence -- will not only provide favorable answers, but will also lead us to a unified general framework that is powerful enough to account for peculiar RG flows and predict new physical phenomena. Namely, using Bott's version of Morse theory we relate the topology of conformal manifolds to certain properties of RG flows that can be used as precise diagnostics and "topological obstructions" for the strong form of the C-theorem in any dimension. Moreover, this framework suggests a precise mechanism for how the violation of the strong C-theorem happens and predicts "phase transitions" along the RG flow when the topological obstruction is non-trivial. Along the way, we also find new conformal manifolds in well-known 4d CFT's and point out connections with the superconformal index and classifying spaces of global symmetry groups.

  6. Sickle erythrocytes inhibit human endothelial cell DNA synthesis

    SciTech Connect

    Weinstein, R.; Zhou, M.A.; Bartlett-Pandite, A.; Wenc, K. )

    1990-11-15

    Patients with sickle cell anemia experience severe vascular occlusive phenomena including acute pain crisis and cerebral infarction. Obstruction occurs at both the microvascular and the arterial level, and the clinical presentation of vascular events is heterogeneous, suggesting a complex etiology. Interaction between sickle erythrocytes and the endothelium may contribute to vascular occlusion due to alteration of endothelial function. To investigate this hypothesis, human vascular endothelial cells were overlaid with sickle or normal erythrocytes and stimulated to synthesize DNA. The erythrocytes were sedimented onto replicate monolayers by centrifugation for 10 minutes at 17 g to insure contact with the endothelial cells. Incorporation of 3H-thymidine into endothelial cell DNA was markedly inhibited during contact with sickle erythrocytes. This inhibitory effect was enhanced more than twofold when autologous sickle plasma was present during endothelial cell labeling. Normal erythrocytes, with or without autologous plasma, had a modest effect on endothelial cell DNA synthesis. When sickle erythrocytes in autologous sickle plasma were applied to endothelial monolayers for 1 minute, 10 minutes, or 1 hour and then removed, subsequent DNA synthesis by the endothelial cells was inhibited by 30% to 40%. Although adherence of sickle erythrocytes to the endothelial monolayers was observed under these experimental conditions, the effect of sickle erythrocytes on endothelial DNA synthesis occurred in the absence of significant adherence. Hence, human endothelial cell DNA synthesis is partially inhibited by contact with sickle erythrocytes. The inhibitory effect of sickle erythrocytes occurs during a brief (1 minute) contact with the endothelial monolayers, and persists for at least 6 hours of 3H-thymidine labeling.

  7. Human Erythrocytes Selectively Bind and Enrich Infectious HIV-1 Virions

    PubMed Central

    Beck, Zoltan; Brown, Bruce K.; Wieczorek, Lindsay; Peachman, Kristina K.; Matyas, Gary R.; Polonis, Victoria R.; Rao, Mangala; Alving, Carl R.

    2009-01-01

    Although CD4(+) cells represent the major target for HIV infection in blood, claims of complement-independent binding of HIV-1 to erythrocytes and the possible role of Duffy blood group antigen, have generated controversy. To examine the question of binding to erythrocytes, HIV-1 was incubated in vitro with erythrocytes from 30 healthy leukapheresis donors, and binding was determined by p24 analysis and adsorption of HIV-1 with reduction of infectivity for CD4(+) target cells. All of the cells, regardless of blood group type, bound HIV-1 p24. A typical preparation of erythrocytes bound <2.4% of the added p24, but erythrocytes selectively removed essentially all of the viral infectivity as determined by decreased infection of CD4(+) target cells; however, cell-associated HIV-1 was approximately 100-fold more efficient, via trans infection, than unadsorbed virus for infection of CD4(+) cells. All of the bound HIV-1 p24 was released by treatment of the cells with EDTA, and binding was optimized by adding Ca2+ and Mg2+ during the washing of erythrocytes containing bound HIV-1. Although the small number of contaminating leukocytes in the erythrocyte preparation also bound HIV-1 p24, there was no significant binding to CD4, and it thus appears that the binding occurred on leukocytes at non-CD4 sites. Furthermore, binding occurred to erythrocyte ghosts from which contaminating leukocytes had been previously removed. The results demonstrate that erythrocytes incubated in vitro with HIV-1 differentially adsorb all of the infectious HIV-1 virions (as opposed to non-infectious or degraded virions) in the absence of complement and independent of blood group, and binding is dependent on divalent cations. By analogy with HIV-1 bound to DC-SIGN on dendritic cells, erythrocyte-bound HIV-1 might comprise an important surface reservoir for trans infection of permissive cells. PMID:20011536

  8. Erythrocyte sequestration and anemia in severe falciparum malaria. Analysis of acute changes in venous hematocrit using a simple mathematical model.

    PubMed Central

    Davis, T M; Krishna, S; Looareesuwan, S; Supanaranond, W; Pukrittayakamee, S; Attatamsoonthorn, K; White, N J

    1990-01-01

    Microvascular erythrocyte sequestration, the characteristic pathological feature of falciparum malaria, was evaluated using a mathematical model in 46 patients with severe infections. From admission radioisotopic circulating red cell volumes and simultaneous venous hematocrits, the model-derived sequestrum hematocrit (mean [95% confidence limits]: 0.70 [0.43-0.97], n = 29) was twice that of peripheral blood (0.33 [0.30-0.36]). Serial reticulocyte and radiolabeled erythrocyte counts indicated that small numbers of cells enter the circulation during initial therapy. The mean fall in hematocrit over 84 h in 26 nontransfused patients conformed to a three-term equation. A first-order decline (t1/2 2.0 h [0.6-3.4]) suggested an average 7.5% plasma volume expansion through rehydration. A zero-order 6.3% (3.1-9.5) fall (t1/2 25.7 h [21.2-30.2]) occurred contemporaneously with a fall in mean parasitemia from 4.5% (3.6-5.4); from these data the model-derived average sequestered erythrocyte volume (4.8% of the admission hematocrit) was similar to the peripheral parasite burden. A second, first-order fall (t1/2 1,047 h [278-1,816]) indicated loss of uninfected erythrocytes with mean lifespan 62 d. Predicted total plasma volume expansion during initial therapy (21.2%) was similar to radioisotopic estimates in 11 patients (17.3% [2.0-33.1]). Application of the model to individual patient data showed wide variations in relative proportions of circulating and sequestered parasitized cells. The model provides evidence of the nature and fate of all parasitized erythrocytes in malaria. PMID:2203822

  9. Oxidative stress in sepsis. Possible production of free radicals through an erythrocyte-mediated positive feedback mechanism.

    PubMed

    Oliveira, Yanaihara Pinchemel Amorim de; Pontes-de-Carvalho, Lain Carlos; Couto, Ricardo David; Noronha-Dutra, Alberto Augusto

    Sepsis is an illness with a high morbidity for which no effective treatment exists. Its treatment has a high cost because it usually requires an intensive care unit and expensive antibiotics. The present study focus in the production of reactive oxygen species in the early stages of sepsis. This study aimed at investigating the production of reactive oxygen specie during the inflammatory response in patients with sepsis. Reactive oxygen specie production and insoluble myeloperoxidase obtained from fresh whole blood were measured by photon counting chemiluminescence in the blood of 18 septic patients and 12 healthy individuals. Modified red blood cells were evaluated by staining of blood smears. The production of reactive oxygen species by macrophages and polymorphonuclear leukocytes put into contact with modified red blood cells were also assessed by photon counting chemiluminescence. The appearance of oxidatively modified erythrocytes, which is an evidence of oxidative stress, was supported by the detection of reactive oxygen species and insoluble myeloperoxidase in the whole blood of all septic patients. Peroxynitrite was the main reactive oxygen species found in the whole blood. Oxidatively modified erythrocytes activated phagocytic cells in vitro, leading to the considerable production of free radicals. It was found that sepsis led to a high oxidative stress and to extensive modification of erythrocytes. It is proposed that a positive feedback mechanism, involving the activation of circulating leukocytes by these modified erythrocytes would maintain the pro-oxidative state even after the disappearance of bacteria. Copyright © 2016 Sociedade Brasileira de Infectologia. Published by Elsevier Editora Ltda. All rights reserved.

  10. Detection of micronucleus and abnormal nucleus in erythrocytes from the gill and kidney of Labeo bata cultivated in sewage-fed fish farms.

    PubMed

    Talapatra, S N; Banerjee, S K

    2007-02-01

    Determination of genotoxic effect in fish, micronucleus test as well as the study of the abnormal shape of nuclei is a suitable measure, in which the presence or absence of genotoxins can be detected in water. In the present study, micronuclei and abnormal nuclei frequencies were scored in the gill and kidney erythrocytes of the fish Labeo bata grown in the sewage-fed fish farms of East Calcutta wetlands. Three experimental sites were chosen, namely, Bantala, Chowbaga and Chingrihata (basically these sites have sewage-fed fishponds), which were compared with fishponds of no sewage influence as the control area. Highly significant differences (P<0.001) were noticed for micronucleus frequencies in the gill and kidney erythrocytes of experimental fishes, where kidney erythrocytes showed an increased value than gill erythrocytes without any statistical differences. The frequencies of nuclear abnormalities such as necrotic cells, apoptotic cells, notch nucleated cells and binucleated cells were also counted separately for gill and kidney erythrocytes, in which significantly (P<0.001, P<0.01, P<0.05) increased values were obtained in comparison to control populations. These genotoxicity results confirmed that the sewage-fed ponds contain genotoxic metals such as Cr, Zn, Cu, Pb, Mn, Fe through wastewater and sludge because of the direct use of sewage water without pretreatment which may lead to health risks among humans through chronic consumption of fish from these experimental fish ponds. Other vertebrates grown in sewage-fed ponds may also suffer a certain amount of genotoxic substances.

  11. Counting coalescent histories.

    PubMed

    Rosenberg, Noah A

    2007-04-01

    Given a species tree and a gene tree, a valid coalescent history is a list of the branches of the species tree on which coalescences in the gene tree take place. I develop a recursion for the number of valid coalescent histories that exist for an arbitrary gene tree/species tree pair, when one gene lineage is studied per species. The result is obtained by defining a concept of m-extended coalescent histories, enumerating and counting these histories, and taking the special case of m = 1. As a sum over valid coalescent histories appears in a formula for the probability that a random gene tree evolving along the branches of a fixed species tree has a specified labeled topology, the enumeration of valid coalescent histories can considerably reduce the effort required for evaluating this formula.

  12. Erythrocyte 22Na+ influx in hypertension

    SciTech Connect

    Shalev, O.; Eaton, J.W.; Ben-Ishay, D.

    1984-01-01

    We assessed 22Na+ uptake by erythrocytes (RBC) from 38 individuals with essential hypertension and 37 healthy controls. All subjects were male, white, non-obese and with normal renal function, obviating sex, race, hormonal, ponderal and renal factors known to influence RBC Na+ handling. The mean +/- sem 22Na+ uptake of the patients was 284 +/- 16 mumole/liter RBC/hour while that of normal controls was 249 +/- 11 mumole/liter RBC/hour; although the difference reached borderline significance, individual values showed considerable overlap. Consequently, in our population, RBC 22Na+ uptake is not a reliable marker for essential hypertension. We believe that previous studies should be reassessed with regard to patients' characteristics and future studies employ rigorous criteria in selection of subjects.

  13. Homeostasis of Extracellular ATP in Human Erythrocytes*

    PubMed Central

    Montalbetti, Nicolas; Leal Denis, Maria F.; Pignataro, Omar P.; Kobatake, Eiry; Lazarowski, Eduardo R.; Schwarzbaum, Pablo J.

    2011-01-01

    We explored the intra- and extracellular processes governing the kinetics of extracellular ATP (ATPe) in human erythrocytes stimulated with agents that increase cAMP. Using the luciferin-luciferase reaction in off-line luminometry we found both direct adenylyl cyclase activation by forskolin and indirect activation through β-adrenergic stimulation with isoproterenol-enhanced [ATP]e in a concentration-dependent manner. A mixture (3V) containing a combination of these agents and the phosphodiesterase inhibitor papaverine activated ATP release, leading to a 3-fold increase in [ATP]e, and caused increases in cAMP concentration (3-fold for forskolin + papaverine, and 10-fold for 3V). The pannexin 1 inhibitor carbenoxolone and a pannexin 1 blocking peptide (10Panx1) decreased [ATP]e by 75–84%. The residual efflux of ATP resulted from unavoidable mechanical perturbations stimulating a novel, carbenoxolone-insensitive pathway. In real-time luminometry experiments using soluble luciferase, addition of 3V led to an acute increase in [ATP]e to a constant value of ∼1 pmol × (106 cells)−1. A similar treatment using a surface attached luciferase (proA-luc) triggered a rapid accumulation of surface ATP levels to a peak concentration of 2.4 pmol × (106 cells)−1, followed by a slower exponential decay (t½ = 3.7 min) to a constant value of 1.3 pmol × (106 cells)−1. Both for soluble luciferase and proA-luc, ATP efflux was fully blocked by carbenoxolone, pointing to a 3V-induced mechanism of ATP release mediated by pannexin 1. Ecto-ATPase activity was extremely low (∼28 fmol × (106 cells min)−1), but nevertheless physiologically relevant considering the high density of erythrocytes in human blood. PMID:21921036

  14. Focusing and alignment of erythrocytes in a viscoelastic medium

    NASA Astrophysics Data System (ADS)

    Go, Taesik; Byeon, Hyeokjun; Lee, Sang Joon

    2017-01-01

    Viscoelastic fluid flow-induced cross-streamline migration has recently received considerable attention because this process provides simple focusing and alignment over a wide range of flow rates. The lateral migration of particles depends on the channel geometry and physicochemical properties of particles. In this study, digital in-line holographic microscopy (DIHM) is employed to investigate the lateral migration of human erythrocytes induced by viscoelastic fluid flow in a rectangular microchannel. DIHM provides 3D spatial distributions of particles and information on particle orientation in the microchannel. The elastic forces generated in the pressure-driven flows of a viscoelastic fluid push suspended particles away from the walls and enforce erythrocytes to have a fixed orientation. Blood cell deformability influences the lateral focusing and fixed orientation in the microchannel. Different from rigid spheres and hardened erythrocytes, deformable normal erythrocytes disperse from the channel center plane, as the flow rate increases. Furthermore, normal erythrocytes have a higher angle of inclination than hardened erythrocytes in the region near the side-walls of the channel. These results may guide the label-free diagnosis of hematological diseases caused by abnormal erythrocyte deformability.

  15. Focusing and alignment of erythrocytes in a viscoelastic medium

    PubMed Central

    Go, Taesik; Byeon, Hyeokjun; Lee, Sang Joon

    2017-01-01

    Viscoelastic fluid flow-induced cross-streamline migration has recently received considerable attention because this process provides simple focusing and alignment over a wide range of flow rates. The lateral migration of particles depends on the channel geometry and physicochemical properties of particles. In this study, digital in-line holographic microscopy (DIHM) is employed to investigate the lateral migration of human erythrocytes induced by viscoelastic fluid flow in a rectangular microchannel. DIHM provides 3D spatial distributions of particles and information on particle orientation in the microchannel. The elastic forces generated in the pressure-driven flows of a viscoelastic fluid push suspended particles away from the walls and enforce erythrocytes to have a fixed orientation. Blood cell deformability influences the lateral focusing and fixed orientation in the microchannel. Different from rigid spheres and hardened erythrocytes, deformable normal erythrocytes disperse from the channel center plane, as the flow rate increases. Furthermore, normal erythrocytes have a higher angle of inclination than hardened erythrocytes in the region near the side-walls of the channel. These results may guide the label-free diagnosis of hematological diseases caused by abnormal erythrocyte deformability. PMID:28117428

  16. The erythrocyte sodium and potassium in patients treated with digoxin.

    PubMed Central

    Morgan, D B; Cumberbatch, M; Cohn, S; Scott, D; Gunasuntharam, T; Davidson, C; Chapman, C

    1980-01-01

    1 Four healthy persons and ten patients with heart failure were studied for 5 to 20 days after they started taking digoxin. The sodium content of their erythrocytes increased and there was an equimolar decrease in potassium content. 2 The increase in erythrocyte sodium for a given increase in plasma digoxin during this acute digitalization was less on average and varied more in the patients than in the healthy persons, that is the patients' erythrocytes were less responsive to digoxin. 3 The average erythrocyte sodium was greater in 183 patients who had been taking digoxin for at least 2 months than in 100 healthy persons not taking digoxin but there was no significant correlation between the plasma digoxin concentrations and erythrocyte sodium concentration in the patients. Indeed, there was no apparent change in the erythrocyte sodium in many of the patients taking digoxin. 4 If the erythrocyte sodium concentration is a reliable guide to the tissue effects of digoxin then the results suggest that there is a wide variation in the response to digoxin between patients both during acute digitalization and during chronic treatment with digoxin. PMID:7426274

  17. Simulation of the osmosis-based drug encapsulation in erythrocytes.

    PubMed

    Ge, Duobiao; Zou, Lili; Li, Chengpan; Liu, Sen; Li, Shibo; Sun, Sijie; Ding, Weiping

    2017-09-20

    Drug-loaded erythrocytes have been proposed for the treatment of disease. A common way to load drugs into erythrocytes is to apply osmotic shock. Currently, osmosis-based drug encapsulation is studied mainly experimentally, whereas a related theoretical model is still incomplete. In this study, a set of equations is developed to simulate the osmosis-based drug-encapsulation process. First, the modeling is validated with hemolysis rates and the drug-loaded quantities to be found in the literature. Then, the variation of the erythrocyte volume, formation of the pore on the erythrocyte membrane, and quantities of drug loaded into and hemoglobin released from erythrocytes are studied. Finally, an optimized operating condition for encapsulating drugs is proposed. The results show that the volume of erythrocytes exposed to hypotonic NaCl solution increases first and then abruptly decreases because of the pore formation; afterwards, it again increases and then decreases slowly. In the presence of the pore, the drug is loaded by diffusion, whereas the leak-induced convection goes against the loading. For an allowed 45% hemolysis rate, with a 10% hematocrit, the optimized NaCl concentration is 0.44%, the optimized time for sealing the loaded erythrocytes with hypertonic NaCl solution is at 6.5 s, and the quantity of albumin (drug) loaded is 4.5 mg/ml cells.

  18. Abnormal erythrocyte membrane protein pattern in severe megaloblastic anemia.

    PubMed Central

    Ballas, S K

    1978-01-01

    The erythrocyte membrane protein pattern of patients with megaloblastic anemia was determined by polyacrylamide gel electrophoresis in sodium dodecyl sulfate. In severe megaloblastic anemia, secondary either to folic acid or vitamin B12 deficiency, the erythrocyte membrane protein pattern was grossly abnormal, lacking bands 1, 2 (spectrin), and 3 and having several diffuse, faster migrating bands. After adequate vitamin replacement therapy, the erythrocyte membrane protein pattern returned to normal. In mild megaloblastic anemia, secondary either to folic acid of vitamin B12 deficiency, and in severe iron deficiency anemia, the erythrocyte membrane protein pattern was normal. Erythrocyte membrane protein pattern of normal membranes did not change after mixing with abnormal membranes before polyacrylamide gel electrophoresis in sodium dodecyl sulfate. Protease activity extracted from membranes of megalocytes was not different from normal. These findings indicate that the erythrocyte membrane protein pattern is abnormal in severe megaloblastic anemia and that this abnormality is not secondary to increased activity of the endogenous erythrocyte membrane proteinase. Images PMID:659579

  19. Erythrocyte hemodynamics in stenotic microvessels: A numerical investigation

    NASA Astrophysics Data System (ADS)

    Wang, T.; Xing, Z. W.

    2013-10-01

    This paper presents a two-dimensional numerical investigation of deformation and motion of erythrocytes in stenotic microvessels using the immersed boundary-fictitious domain method. The erythrocytes were modeled as biconcave-shaped closed membranes filled with cytoplasm. We studied the biophysical characteristics of human erythrocytes traversing constricted microchannels with the narrowest cross-sectional diameter as small as 3 μm. The effects of essential parameters, namely, stenosis severity, shape of the erythrocytes, and erythrocyte membrane stiffness, were simulated and analyzed in this study. Moreover, simulations were performed to discuss conditions associated with the shape transitions of the cells along with the relative effects of radial position and initial orientation of erythrocytes, membrane stiffness, and plasma environments. The simulation results were compared with existing experiment findings whenever possible, and the physical insights obtained were discussed. The proposed model successfully simulated rheological behaviors of erythrocytes in microscale flow and thus is applicable to a large class of problems involving fluid flow with complex geometry and fluid-cell interactions. Our study would be helpful for further understanding of pathology of malaria and some other blood disorders.

  20. Erythrocyte hemodynamics in stenotic microvessels: A numerical investigation

    NASA Astrophysics Data System (ADS)

    Wang, Tong; Xing, Zhongwen

    2014-03-01

    This paper presents a two-dimensional numerical investigation of deformation and motion of erythrocytes in stenotic microvessels using the immersed boundary-fictitious domain method. The erythrocytes were modeled as biconcave-shaped closed membranes filled with cytoplasm. We studied the biophysical characteristics of human erythrocytes traversing constricted microchannels with the narrowest cross-sectional diameter as small as 3 μm. The effects of essential parameters, namely, stenosis severity, shape of the erythrocytes, and erythrocyte membrane stiffness, were simulated and analyzed in this study. Moreover, simulations were performed to discuss conditions associated with the shape transitions of the cells along with the relative effects of radial position and initial orientation of erythrocytes, membrane stiffness, and plasma environments. The simulation results were compared with existing experiment findings whenever possible, and the physical insights obtained were discussed. The proposed model successfully simulated rheological behaviors of erythrocytes in microscale flow and thus is applicable to a large class of problems involving fluid flow with complex geometry and fluid-cell interactions. Our study would be helpful for further understanding of pathology of malaria and some other blood disorders.

  1. Erythrocyte hemodynamics in stenotic microvessels: a numerical investigation.

    PubMed

    Wang, T; Xing, Z W

    2013-10-01

    This paper presents a two-dimensional numerical investigation of deformation and motion of erythrocytes in stenotic microvessels using the immersed boundary-fictitious domain method. The erythrocytes were modeled as biconcave-shaped closed membranes filled with cytoplasm. We studied the biophysical characteristics of human erythrocytes traversing constricted microchannels with the narrowest cross-sectional diameter as small as 3 μm. The effects of essential parameters, namely, stenosis severity, shape of the erythrocytes, and erythrocyte membrane stiffness, were simulated and analyzed in this study. Moreover, simulations were performed to discuss conditions associated with the shape transitions of the cells along with the relative effects of radial position and initial orientation of erythrocytes, membrane stiffness, and plasma environments. The simulation results were compared with existing experiment findings whenever possible, and the physical insights obtained were discussed. The proposed model successfully simulated rheological behaviors of erythrocytes in microscale flow and thus is applicable to a large class of problems involving fluid flow with complex geometry and fluid-cell interactions. Our study would be helpful for further understanding of pathology of malaria and some other blood disorders.

  2. In Vitro Protective Effect of Phikud Navakot Extraction on Erythrocyte

    PubMed Central

    2016-01-01

    Phikud Navakot (PN), Thai herbal remedy in National List of Essential Medicines, has been claimed to reduce many cardiovascular symptoms especially dizziness and fainting. Apart from blood supply, erythrocyte morphology, in both shape and size, is one of the main consideration factors in cardiovascular diseases and may be affected by vascular oxidative stress. However, little is known about antioxidative property of PN on erythrocyte to preserve red blood cell integrity. In this study, 1,000 μM hydrogen peroxide-induced oxidative stress was conducted on sheep erythrocyte. Three doses of PN (1, 0.5, and 0.25 mg/mL) and 10 μM of ascorbic acid were compared. The released hemoglobin absorbance was measured to demonstrate hemolysis. Electron microscopic and immunohistochemical studies were also performed to characterize dysmorphic erythrocyte and osmotic ability in relation to aquaporin- (AQP-) 1 expression, respectively. The results revealed that all doses of PN and ascorbic acid decreased the severity of dysmorphic erythrocyte, particularly echinocyte, acanthocyte, knizocyte, codocyte, clumping, and other malformations. However, the most effective was 0.5 mg/mL PN dosage. In addition, hydrostatic pressure may be increased in dysmorphic erythrocyte in association with AQP-1 upregulation. Our results demonstrated that PN composes antioxidative effect to maintain the integrity and osmotic ability on sheep erythrocyte. PMID:28003847

  3. Interaction between plant polyphenols and the erythrocyte membrane.

    PubMed

    Cyboran, Sylwia; Oszmiański, Jan; Kleszczyńska, Halina

    2012-03-01

    The purpose of these studies was to determine the effect of polyphenols contained in extracts from apple, strawberry and blackcurrant on the properties of the erythrocyte membrane, treated as a model of the biological membrane. To this end, the effect of the substances used on hemolysis, osmotic resistance and shape of erythrocytes, and on packing order in the hydrophilic region of the erythrocyte membrane was studied. The investigation was performed with spectrophotometric and fluorimetric methods, and using the optical microscope. The hemolytic studies have shown that the extracts do not induce hemolysis at the concentrations used. The results obtained from the spectrophotometric measurements of osmotic resistance of erythrocytes showed that the polyphenols contained in the extracts cause an increase in the resistance, rendering them less prone to hemolysis in hypotonic solutions of sodium chloride. The fluorimetric studies indicate that the used substances cause a decrease of packing order in the hydrophilic area of membrane lipids. The observations of erythrocyte shapes in a biological optical microscope have shown that, as a result of the substances' action, the erythrocytes become mostly echinocytes, which means that the polyphenols of the extracts localize in the outer lipid monolayer of the erythrocyte membrane. The results obtained indicate that, in the concentration range used, the plant extracts are incorporated into the hydrophilic area of the membrane, modifying its properties.

  4. Erythrocyte survival studies in a rat myelogenous leukemia

    SciTech Connect

    Derelanko, M.J.; Meagher, R.C.; Lobue, J.; Khouri, J.A.; Gordon, A.S.

    1982-11-01

    To determine the extent intrinsic erythrocyte defects and/or extrinsic factors were involved in anemia of rats bearing Shay chloroleukemia (SCL), survival of /sup 3/H-DFP labeled erythrocytes was studied in leukemic and nonleukemic hosts. Red blood cells labeled before induction of leukemia, were rapidly lost from the peripheral circulation of SCL rats in terminal stages of disease. However, labeled erythrocytes from terminal SCL animals displayed normal lifespans when transfused into nonleukemic controls. Thus the anemia of this leukemia probably resulted from extrinsic factors associated with the leukemic process. Hemorrhage appeared to be primarily responsible for the anemia of this disease.

  5. Mature Erythrocytes of Iguana iguana (Squamata, Iguanidae) Possess Functional Mitochondria.

    PubMed

    Di Giacomo, Giuseppina; Campello, Silvia; Corrado, Mauro; Di Giambattista, Livia; Cirotti, Claudia; Filomeni, Giuseppe; Gentile, Gabriele

    2015-01-01

    Electron microscopy analyses of Iguana iguana blood preparations revealed the presence of mitochondria within erythrocytes with well-structured cristae. Fluorescence microscopy analyses upon incubation with phalloidin-FITC, Hoechst 33342 and mitochondrial transmembrane potential (Δψm)-sensitive probe MitoTracker Red indicated that mitochondria i) widely occur in erythrocytes, ii) are polarized, and iii) seem to be preferentially confined at a "perinuclear" region, as confirmed by electron microscopy. The analysis of NADH-dependent oxygen consumption showed that red blood cells retain the capability to consume oxygen, thereby providing compelling evidence that mitochondria of Iguana erythrocytes are functional and capable to perform oxidative phosphorylation.

  6. Mature Erythrocytes of Iguana iguana (Squamata, Iguanidae) Possess Functional Mitochondria

    PubMed Central

    Di Giacomo, Giuseppina; Campello, Silvia; Corrado, Mauro; Di Giambattista, Livia; Cirotti, Claudia; Filomeni, Giuseppe; Gentile, Gabriele

    2015-01-01

    Electron microscopy analyses of Iguana iguana blood preparations revealed the presence of mitochondria within erythrocytes with well-structured cristae. Fluorescence microscopy analyses upon incubation with phalloidin-FITC, Hoechst 33342 and mitochondrial transmembrane potential (Δψm)-sensitive probe MitoTracker Red indicated that mitochondria i) widely occur in erythrocytes, ii) are polarized, and iii) seem to be preferentially confined at a "perinuclear" region, as confirmed by electron microscopy. The analysis of NADH-dependent oxygen consumption showed that red blood cells retain the capability to consume oxygen, thereby providing compelling evidence that mitochondria of Iguana erythrocytes are functional and capable to perform oxidative phosphorylation. PMID:26367118

  7. Attachment of killed Mycoplasma gallisepticum cells and membranes to erythrocytes

    SciTech Connect

    Banai, M.; Kahane, I.; Feldner, J.; Razin, S.

    1981-11-01

    To correlate viability with attachment capacity, Mycoplasma gallisepticum cells harvested at different growth phases and treated by various agents were tested for their capacity to attach to human erythrocytes. The results show that viability per se is not essential for M. gallisepticum attachment to erythrocytes, as cells killed by ultraviolet irradiation and membranes isolated by lysing M. gallisepticum cells by various means retained attachment capacity. However, treatment of the mycoplasmas by protein-denaturing agents, such as heart, glutaraldehyde, or prolonged exposure to low pH, drastically affected or even abolished attachment, supporting the protein nature of the mycoplasma membrane components responsible for specific binding to the sialoglycoprotein receptors on the erythrocytes.

  8. [Erythrocyte alloimmunization in pregnant women, clinical importance and laboratory diagnostics].

    PubMed

    Holusková, I; Lubušký, M; Studničková, M; Procházka, M

    2013-01-01

    The aim of this review is to give comprehensive summary of erythrocyte alloimunization of pregnant women, laboratory dignostics and clinical importance. Review. University Hospital Olomouc, Transfusion Department, Department of Obstetrics and Gynecology. Based on literature analysis using database search engines PubMed, Google Scholar, Ovid in field of erythrocyte antibodies, laboratory diagnostics and clinical importance up-to-date knowledge. Erythrocyte alloimunization anti-D antibodies decreases in connection with the introduction of immunoprofylaxis. Immunization of non RhD antibodies with impossibility using of immunoprofylaxis remains still clinical problem.

  9. Forssman-like activity of different teleost and anuran erythrocytes

    PubMed Central

    Chuba, J. V.; Kuhns, W. J.; Nigrelli, R. F.

    1971-01-01

    The comparative agglutination of sheep, teleost and anuran erythrocytes with selected human sera was studied before and after absorption with boiled guinea-pig kidney antigen. Among frogs of the Rana genus, only bullfrog (R. catesbiana) erythrocytes displayed Forssman-like activity. The previously unstudied erythrocytes of several species of teleosts also displayed Forssman-like activity. The concept is advanced that haptenic heteroraccharides with varying degrees of structural similarity and `blood group' activity are ubiquitously distributed among the cell-membrane receptors of phylogenetically diverse species of animals. PMID:5553070

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

    PubMed

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

    2014-01-01

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

  11. Human erythrocyte antigens. Regulation of expression of a novel erythrocyte surface antigen by the inhibitor Lutheran In(Lu) gene.

    PubMed Central

    Telen, M J; Eisenbarth, G S; Haynes, B F

    1983-01-01

    Our study describes a novel human erythrocyte protein antigen, the expression of which is regulated by the rare Lutheran inhibitor In(Lu) gene. We have produced a monoclonal antibody (A3D8) that bound strongly to erythrocytes from subjects with Lutheran phenotypes Lu(a+b+), Lu(a+b-), and Lu(a-b+) but bound negligibly to erythrocytes from subjects with the dominant form of Lu(a-b-) phenotype, reflecting inheritance of the In(Lu) gene. Importantly, erythrocytes from an individual with the recessive form of Lu(a-b-) phenotype (i.e., absence of the In(Lu) gene and absence of genes encoding for Lutheran antigens) showed reactivity with A3D8 antibody comparable to that seen with Lu(a+) or Lu(b+) erythrocytes. A3D8 antigen activity was also found on all leukocytes and in serum and plasma; this activity also appeared to be regulated by the In(Lu) gene in serum, plasma, and on a subset of leukocytes. Thus, we have identified a human erythrocyte protein whose expression is modified by the In(Lu) gene. This knowledge that such an antigen exists on erythrocytes and in normal plasma should allow further studies into the molecular genetics of the In(Lu) gene and into the functional and structural significance of the A3D8 antigen. PMID:6863545

  12. Effect of donor age on the susceptibility of erythrocytes and erythrocyte membranes to cumene hydroperoxide-induced oxidative stress.

    PubMed

    Onaran, I; Yalçin, A S; Sultuybek, G

    1997-11-01

    A comparative study on erythrocytes and erythrocyte membranes of healthy elderly and young adults was carried out to understand how the antioxidant defense capacity is effected by aging. The levels of endogenous malondialdehyde and Ca(2+)-ATPase activity were taken as indices of oxidative damage. In addition, chemiluminescence measurements were performed on intact erythrocytes. The susceptibility of these parameters to in vitro cumene hydroperoxide, under low oxidant level that does not induce hemolysis, was also taken as an age-related indicator of the endogenous peroxidative potential of the erythrocytes. Our data showed that the content of malondialdehyde and Ca(2+)-ATPase activity did not change with age. Furthermore, the susceptibility of intact erythrocytes to oxidative stress did not change in the elderly group. However, under the same conditions erythrocyte membranes were more susceptible to oxidative damage in the elderly than young adults. Our results also showed that antioxidant defenses were overwhelmed in intact erythrocytes of the elderly at high concentrations of cumene hydroperoxide.

  13. Hemorheological changes and hematometric erythrocyte characteristics in rats after sodium nitrite intoxication

    NASA Astrophysics Data System (ADS)

    Ivanov, Ivan; Gluhcheva, Yordanka; Petrova, Emilia; Antonova, Nadia

    2014-05-01

    Sodium nitrite (NaNO2) is a precursor to a variety of organic compounds (pharmaceuticals, dyes and pesticides), but it is best known as a food additive. The aim of the study is to investigate the influence of acute (i.p.) treatment of Wistar rats with NaNO2 (at the dose of 50 mg/kg b.w.) on the blood rheological properties and erythrocyte hematometric indices (Hb, HCT, RBC, MCV, RDW, MCH, MCHC). The significant differences were not found in the whole blood viscosity (WBV) values of the control and NaNO2-treated groups. The changes in the erythrocyte hematometric indices were statistically significant for RDW, MCHC and MCH at the 1st hour, five- and ten days after NaNO2 administration. Interestingly, at the day 5th of the NaNO2 treatment we obtained statistically significant lower values for the RBC count, Hb, HCT, RDW, as well as elevated indices MCV (no statistically significant), MCH, MCHC. The results obtained indicate that hemorheological and hematometric parameters examined should be monitored in cases of acute exposure to nitrites — for the purposes of clinical toxicology. The quantitative values of hematometric indices reported in our experimental model could be suitable for predicting NaNO2 intoxication and methemoglobinemia in animals and humans.

  14. Flow cytometric scoring of micronucleated erythrocytes: an efficient platform for assessing in vivo cytogenetic damage.

    PubMed

    Dertinger, Stephen D; Torous, Dorothea K; Hayashi, Makoto; MacGregor, James T

    2011-01-01

    The relative simplicity of the micronucleated erythrocyte endpoint has made it amenable to automated scoring approaches. Flow cytometry is one such scoring platform that has been employed successfully. This review describes the evolution and properties of flow cytometry-based scoring of micronucleated erythrocytes. The methodology has become widely applied to rodent blood specimens and the high throughput nature of the technology provides a number of advantages over manual microscopic scoring. For instance, the ability to efficiently survey many dose levels and many more cells per specimen relative to microscopy benefits studies that are designed to identify no observable effect levels or lowest observable effect levels. Furthermore, flow cytometry makes it practical to study species with low spontaneous reticulocyte (RET) counts and micronucleus (MN) frequencies, thereby facilitating integration of blood-based micronucleated reticulocyte (MN-RET) frequency measurements into experiments conducted across species of toxicological interest. This capability enhances genotoxicity assessments that have historically been made in dedicated MN tests performed in one species. Importantly, the feasibility of using MN-RET frequencies in blood from humans as an index of genetic damage in bone marrow opens a critical area of application that had not been practical previously. We conclude with recommendations for additional work that is needed to more fully realise the potential of flow cytometric in vivo MN scoring.

  15. "Just" Counting: Young Children's Oral Counting and Enumeration

    ERIC Educational Resources Information Center

    Threlfall, John; Bruce, Bob

    2005-01-01

    This article focuses on the specific skills and abilities of young children in oral counting and enumeration. Responses to an oral counting task and an enumeration task by a sample (n=93) of 3- and 4-year old children attending a range of pre-five establishments in an urban district of northern England are described. The findings, whilst providing…

  16. Kids Count [and] Families Count in Delaware: Fact Book, 1998.

    ERIC Educational Resources Information Center

    Nelson, Carl, Ed.; Wilson, Nancy, Ed.

    This Kids Count report is combined with Families Count, and provides information on statewide trends affecting children and families in Delaware. The first statistical profile is based on 10 main indicators of child well-being: (1) births to teens; (2) low birth weight babies; (3) infant mortality; (4) child deaths; (5) teen deaths; (6) juvenile…

  17. Compton suppression gamma-counting: The effect of count rate

    USGS Publications Warehouse

    Millard, H.T.

    1984-01-01

    Past research has shown that anti-coincidence shielded Ge(Li) spectrometers enhanced the signal-to-background ratios for gamma-photopeaks, which are situated on high Compton backgrounds. Ordinarily, an anti- or non-coincidence spectrum (A) and a coincidence spectrum (C) are collected simultaneously with these systems. To be useful in neutron activation analysis (NAA), the fractions of the photopeak counts routed to the two spectra must be constant from sample to sample to variations must be corrected quantitatively. Most Compton suppression counting has been done at low count rate, but in NAA applications, count rates may be much higher. To operate over the wider dynamic range, the effect of count rate on the ratio of the photopeak counts in the two spectra (A/C) was studied. It was found that as the count rate increases, A/C decreases for gammas not coincident with other gammas from the same decay. For gammas coincident with other gammas, A/C increases to a maximum and then decreases. These results suggest that calibration curves are required to correct photopeak areas so quantitative data can be obtained at higher count rates. ?? 1984.

  18. Subcutaneous administration of carrier erythrocytes: slow release of entrapped agent

    SciTech Connect

    DeLoach, J.R.; Corrier, D.E.

    1988-08-01

    Carrier erythrocytes administered subcutaneously in mice release encapsulated molecules at the injection site and through cells that escape the injection site. One day postinjection, the efflux of encapsulated (/sup 14/C)sucrose, (/sup 3/H)inulin, and /sup 51/Cr-hemoglobin from the injection site was 45, 55, and 65%, respectively. Intact carrier erythrocytes escaped the injection site and entered the blood circulation carrying with them the encapsulated molecules. Most of the encapsulated (/sup 3/H)inulin that reached whole blood circulated within erythrocytes. Small but measurable numbers of encapsulated molecules were trapped within lymph nodes. Subcutaneous injection of carrier erythrocytes may allow for limited extravascular tissue targeting of drugs.

  19. Immune Evasion Strategies of Pre-Erythrocytic Malaria Parasites

    PubMed Central

    Zheng, Hong; Tan, Zhangping

    2014-01-01

    Malaria is a mosquito-borne infectious disease of humans. It begins with a bite from an infected female Anopheles mosquito and leads to the development of the pre-erythrocytic and blood stages. Blood-stage infection is the exclusive cause of clinical symptoms of malaria. In contrast, the pre-erythrocytic stage is clinically asymptomatic and could be an excellent target for preventive therapies. Although the robust host immune responses limit the development of the liver stage, malaria parasites have also evolved strategies to suppress host defenses at the pre-erythrocytic stage. This paper reviews the immune evasion strategies of malaria parasites at the pre-erythrocytic stage, which could provide us with potential targets to design prophylactic strategies against malaria. PMID:24891764

  20. Cryo scanning electron microscopy of Plasmodium falciparum-infected erythrocytes.

    PubMed

    Hempel, Casper

    2017-07-01

    Plasmodium falciparum invades erythrocytes as an essential part of their life cycle. While living inside erythrocytes, the parasite remodels the cell's intracellular organization as well as its outer surface. Late trophozoite-stage parasites and schizonts introduce numerous small protrusions on the erythrocyte surface, called knobs. Current methods for studying these knobs include atomic force microscopy and electron microscopy. Standard electron microscopy methods rely on chemical fixation and dehydration modifying cell size. Here, a novel method is presented using rapid freezing and scanning electron microscopy under cryogenic conditions allowing for high resolution and magnification of erythrocytes. This novel technique can be used for precise estimates of knob density and for studies on cytoadhesion. © 2017 APMIS. Published by John Wiley & Sons Ltd.

  1. Preclinical trials of human erythrocyte superoxide dismutase injection.

    PubMed

    Zhou, C; Fang, Y; Jiang, D; Yang, S; Lu, X; Sui, J; Li, P; Ren, J

    2000-07-01

    To assess the quality of human erythrocyte superoxide dismutase (SOD) injection reaching the official standard for its clinical uses. Human erythrocyte SOD injection prepared by McCord-Fridovich's method without column chromatography but with some modifications was used in preclinical trials, to observe the general pharmacology and pharmacodynamics of the product. The quality of human erythrocyte SOD injection conformed to the official standard of a biological product, which was found to be non-toxic and did not have any effects on the central and autonomic nervous systems as well as cardiovascular and respiratory systems. The efficacy of anti-inflammation and promotion of immuno-regulation especially on carrageenan and adjuvant-induced polyarthritis were shown in animals. Human erythrocyte SOD injection is appropriate for prophylactic and therapeutic uses in clinical trials.

  2. Erythrocyte and leukocyte: two partners in bacteria killing.

    PubMed

    Minasyan, Hayk A

    2014-01-01

    Leukocytes can't perform phagocytosis in blood stream. Blood velocity prevents phagocytosis because there is no time for leukocyte to recognize and catch bacteria. Bloodstream clearance from pathogens is performed by erythrocytes. During motion in bloodstream erythrocytes become charged by triboelectric effect. This charge attracts bacteria and fixes them on the surface of erythrocyte, then bacteria are engulfed and killed by hemoglobin oxygen. In bloodstream, leukocyte thin-wrinkled elastic membrane can't be charged by triboelectric effect and so leukocyte can't catch bacteria by means of electrostatic attraction force. Leukocytes engulf and kill bacteria out of blood circulatory system: in tissues, lymph nodes, slow velocity lymph, etc. Erythrocyte and leukocyte are bactericidal partners: the first kills bacteria in bloodstream, the second kills them locally, out of blood circulation.

  3. Purification and characterization of acylphosphatase erythrocyte isoenzyme from turkey muscle.

    PubMed

    Stefani, M; Degl'Innocenti, D; Berti, A; Cappugi, G; Manao, G; Camici, G; Ramponi, G

    1990-10-01

    An acylphosphatase has been purified from turkey muscle in a rapid and high-yield way. The enzyme has been characterized for structural, kinetic, and immunological parameters, as well as with regard to its stability to thermal, urea, and phenylglyoxal inactivation. The enzyme is quite different from the turkey muscular isoenzyme, and shows structural and kinetic properties that are very similar to those previously reported for the erythrocyte isoenzyme from human erythrocytes and from chicken muscle. From the data reported it appears that this enzyme corresponds to the acylphosphatase erythrocyte isoenzyme. Unlike the erythrocyte isoenzymes studied so far, this enzyme is able to cross-react with antibodies that are raised against the muscular isoenzyme.

  4. Hemoglobin C trait accentuates erythrocyte dehydration in hereditary xerocytosis.

    PubMed

    Yang, Elizabeth; Voelkel, Erin B; Lezon-Geyda, Kimberly; Schulz, Vincent P; Gallagher, Patrick G

    2017-08-01

    A 17-year-old male presented with acute hemolysis with stomatocytosis, elevated mean corpuscular hemoglobin concentration (MCHC), and osmotic gradient ektacytometry consistent with marked erythrocyte dehydration. Erythrocytes from both parents also demonstrated evidence of dehydration with elevated MCHC and abnormal ektacytometry, but neither to the degree of the patient. Genetic studies revealed the patient had hereditary xerocytosis (HX) due to a novel PIEZO1 mutation inherited from his mother and hemoglobin C (HbC) trait inherited from his father. HbC trait accentuated the erythrocyte dehydration of HX. Coinheritance of interrelated disorders and/or modifier alleles should be considered whenever severe erythrocyte dehydration is observed. © 2017 Wiley Periodicals, Inc.

  5. Exposure to ozone and erythrocyte osmotic resistance in the rat

    SciTech Connect

    Ikemi, Y.; Ohmori, K.; Ito, T.; Osaka, F.; Matuura, Y. )

    1992-10-01

    In order to learn the biological effect of photochemical oxidants on living bodies, we exposed newborn and adult rats, of both sexes, to ozone at a concentration of 0.25 ppm, which can be encountered in an urban environment, and then measured the osmotic resistance of their erythrocytes. The results of experiments using newborn rats indicated a positive increase in the osmotic resistance of erythrocytes in whole blood following ozone exposure for 4 weeks. An increase in the osmotic resistance of erythrocytes in the top part obtained by centrifugation was observed following ozone exposure for 12 weeks. This tendency was especially evident among male rats. On the other hand, no increase in the osmotic resistance of erythrocytes was recognized in the adult animals which had been exposed to the same concentration of ozone for 18 months.

  6. Permeability of human erythrocyte membrane vesicles to alkali cations.

    PubMed

    Sze, H; Solomon, A K

    1979-02-02

    The permeability of inside-out and right-side-out vesicles from erythrocyte membranes to inorganic cations was determined quantitatively. Using 86Rb as a K analog, we have measured the rate constant of 86Rb efflux from vesicles under equilibrium exchange conditions, using a dialysis procedure. The permeability coefficients of the vesicles to Rb are only about an order of magnitude greater than that of whole erythrocytes. Furthermore, we have measured many of the specialized transport systems known to exist in erythrocytes and have shown that glucose, sulfate, ATP-dependent Ca and ATP-dependent Na transport activities are retained by the vesicle membranes. These results suggest that inside-out and right-side-out vesicles can be used effectively to study transport properties of erythrocyte membranes.

  7. Falciparum Malaria-Infected Erythrocytes Specifically Bind to Cultured Human Endothelial Cells

    NASA Astrophysics Data System (ADS)

    Udeinya, Iroka J.; Schmidt, John A.; Aikawa, Masamichi; Miller, Louis H.; Green, Ira

    1981-07-01

    Erythrocytes infected with the late stages of the human malarial parasite Plasmodium falciparum became attached to a subpopulation of cultured human endothelial cells by knoblike protrusions on the surface of the infected erythrocytes. Infected erythrocytes did not bind to cultured fibroblasts; uninfected erythrocytes did not bind to either endothelial cells or fibroblasts. The results suggest a specific receptor-ligand interaction between endothelial cells and a component, or components, in the knobs of the infected erythrocytes.

  8. Morphological characteristics of blood cells in monitor lizards: is erythrocyte size linked to actual body size?

    PubMed

    Frýdlová, Petra; Hnízdo, Jan; Chylíková, Lenka; Simková, Olga; Cikánová, Veronika; Velenský, Petr; Frynta, Daniel

    2013-04-01

    Blood cell morphology and count are not uniform across species. Recently, between-species comparisons revealed that the size of red blood cells is associated with body size in some lizard taxa, and this finding was interpreted in the context of the metabolic theory. In the present study, we examined the numbers and the size of blood cells in 2 species of monitor lizards, the mangrove-dwelling monitor (Varanus indicus) and the savannah monitor (V. exanthematicus), and we compared these traits in individuals of different body size. The results revealed that during the course of ontogeny, the size of red blood cells increases with body mass. Because the mass-specific metabolic rate decreases with body size and the cell volume-to-surface ratio decreases with the cell size, changes in the erythrocyte size might be the result of oxygen transport adjustment. © 2012 Wiley Publishing Asia Pty Ltd, ISZS and IOZ/CAS.

  9. Proteome analysis of the triton-insoluble erythrocyte membrane skeleton.

    PubMed

    Basu, Avik; Harper, Sandra; Pesciotta, Esther N; Speicher, Kaye D; Chakrabarti, Abhijit; Speicher, David W

    2015-10-14

    Erythrocyte shape and membrane integrity is imparted by the membrane skeleton, which can be isolated as a Triton X-100 insoluble structure that retains the biconcave shape of intact erythrocytes, indicating isolation of essentially intact membrane skeletons. These erythrocyte "Triton Skeletons" have been studied morphologically and biochemically, but unbiased proteome analysis of this substructure of the membrane has not been reported. In this study, different extraction buffers and in-depth proteome analyses were used to more fully define the protein composition of this functionally critical macromolecular complex. As expected, the major, well-characterized membrane skeleton proteins and their associated membrane anchors were recovered in good yield. But surprisingly, a substantial number of additional proteins that are not considered in erythrocyte membrane skeleton models were recovered in high yields, including myosin-9, lipid raft proteins (stomatin, flotillin1 and 2), multiple chaperone proteins (HSPs, protein disulfide isomerase and calnexin), and several other proteins. These results show that the membrane skeleton is substantially more complex than previous biochemical studies indicated, and it apparently has localized regions with unique protein compositions and functions. This comprehensive catalog of the membrane skeleton should lead to new insights into erythrocyte membrane biology and pathogenic mutations that perturb membrane stability. Biological significance Current models of erythrocyte membranes describe fairly simple homogenous structures that are incomplete. Proteome analysis of the erythrocyte membrane skeleton shows that it is quite complex and includes a substantial number of proteins whose roles and locations in the membrane are not well defined. Further elucidation of interactions involving these proteins and definition of microdomains in the membrane that contain these proteins should yield novel insights into how the membrane skeleton

  10. Low toxicity method of inhibiting sickling of sickle erythrocytes

    DOEpatents

    Packer, Lester; Bymun, Edwin N.

    1977-01-01

    A low toxicity method of inhibiting sickling of sickle erythrocytes which comprises intermixing the erythrocytes with an effective anti-sickling amount of a water-soluble imidoester of the formula RC(=NH)OR' wherein R is an alkyl group of 1 - 8 carbon atoms, particularly 1 - 4 carbon atoms, and R' is an alkyl group of 1 - 4 carbon atoms, specifically methyl or ethyl acetimidate.

  11. Kinetics of extracellular ATP in mastoparan 7-activated human erythrocytes

    PubMed Central

    Denis, María Florencia Leal; Incicco, J. Jeremías; Espelt, María Victoria; Verstraeten, Sandra V.; Pignataro, Omar P.; Lazarowski, Eduardo R.; Schwarzbaum, Pablo J.

    2014-01-01

    SUMMARY Background The peptide mastoparan 7 (MST7) stimulated ATP release in human erythrocytes. We explored intra- and extracellular processes governing the time-dependent accumulation of extracellular ATP (i.e., ATPe kinetics). Methods Human erythrocytes were treated with MST7 in the presence or absence of two blockers of pannexin 1. ATPe concentration was monitored by luciferin-luciferase based real-time luminometry. Results Exposure of human erythrocytes to MST7 led to an acute increase in [ATPe], followed by a slower increase phase. ATPe kinetics reflected a strong activation of ATP efflux and a low rate of ATPe hydrolysis by ectoATPase activity. Enhancement of [ATPe] by MST7 required adhesion of erythrocytes to poly-D-lysin-coated coverslips, and correlated with a 31% increase of cAMP and 10% cell swelling. However, when MST7 was dissolved in a hyperosmotic medium to block cell swelling, ATPe accumulation was inhibited by 49%. Erythrocytes pre-exposure to 10 μM of either carbenoxolone or probenecid, two blockers of pannexin 1, exhibited a partial reduction of ATP efflux. Erythrocytes from pannexin 1 knockout mice exhibited similar ATPe kinetics as those of wild type mice erythrocytes exposed to pannexin 1 blockers. Conclusions MST7 induced release of ATP required either cell adhesion or strong activation of cAMP synthesis. Part of this release required cell swelling. Kinetic analysis and a data driven model suggested that ATP efflux is mediated by two ATP conduits displaying different kinetics, with one conduit being fully blocked by pannexin 1 blockers. General Significance Kinetic analysis of extracellular ATP accumulation from human erythrocytes and potential effects on microcirculation. PMID:23742824

  12. [Comparative estimation of morphofunction characteristics alive and fixed erythrocytes].

    PubMed

    Skorkina, M Iu; Fedorova, M Z; Cherniavskikh, S D; Zabiniakov, N A; Sladkova, E A

    2011-01-01

    The method of power spectroscopy carries out the quantitative analysis of elastic properties of alive cells. It has been established that the highest indicators of elasticity nuclear (amphibious) and denuclearized (mammals) alive erythrocytes are registered in epi- and perinuclear space. When fixing with methanol and drying of cells the greatest values of the elastic modulus are displaced to the periphery of the cells. The revealed inversion of elasticity properties of erythrocytes must be considered when assessing the morphofunction characteristics of the fixed cells.

  13. Amodiaquine failure associated with erythrocytic glutathione in Plasmodium falciparum malaria

    PubMed Central

    Zuluaga, Lina; Pabón, Adriana; López, Carlos; Ochoa, Aleida; Blair, Silvia

    2007-01-01

    Objective To establish the relationship between production of glutathione and the therapeutic response to amodiaquine (AQ) monotherapy in Plasmodium falciparum non-complicated malaria patients. Methodology Therapeutic response to AQ was evaluated in 32 patients with falciparum malaria in two townships of Antioquia, Colombia, and followed-up for 28 days. For every patient, total glutathione and enzymatic activity (glutathione reductase, GR, and γ-glutamylcysteine synthetase, γ-GCS) were determined in parasitized erythrocytes, non-infected erythrocytes and free parasites, on the starting day (day zero, before ingestion of AQ) and on the day of failure (in case of occurrence). Results There was found an AQ failure of 31.25%. Independent of the therapeutic response, on the starting day and on the day of failure, lower total glutathione concentration and higher GR activities in parasitized erythrocytes were found, compared with non-infected erythrocytes (p < 0.003). In addition, only on the day of failure, γ-GCS activity of parasitized erythrocytes was higher, compared with that of healthy erythrocytes (p = 0.01). Parasitized and non-parasitized erythrocytes in therapeutic failure patients (TF) had higher total glutathione on the starting day compared with those of adequate clinical response (ACR) (p < 0.02). Parasitized erythrocytes of TF patients showed lower total glutathione on the failure day, compared with starting day (p = 0.017). No differences was seen in the GR and γ-GCS activities by compartment, neither between the two therapeutic response groups nor between the two treatment days. Conclusion This study is a first approach to explaining P. falciparum therapeutic failure in humans through differences in glutathione metabolism in TF and ACR patients. These results suggest a role for glutathione in the therapeutic failure to antimalarials. PMID:17451604

  14. Streptococcus pneumoniae Invades Erythrocytes and Utilizes Them to Evade Human Innate Immunity

    PubMed Central

    Yamaguchi, Masaya; Terao, Yutaka; Mori-Yamaguchi, Yuka; Domon, Hisanori; Sakaue, Yuuki; Yagi, Tetsuya; Nishino, Kunihiko; Yamaguchi, Akihito; Nizet, Victor; Kawabata, Shigetada

    2013-01-01

    Streptococcus pneumoniae, a Gram-positive bacterium, is a major cause of invasive infection-related diseases such as pneumonia and sepsis. In blood, erythrocytes are considered to be an important factor for bacterial growth, as they contain abundant nutrients. However, the relationship between S. pneumoniae and erythrocytes remains unclear. We analyzed interactions between S. pneumoniae and erythrocytes, and found that iron ion present in human erythrocytes supported the growth of Staphylococcus aureus, another major Gram-positive sepsis pathogen, while it partially inhibited pneumococcal growth by generating free radicals. S. pneumoniae cells incubated with human erythrocytes or blood were subjected to scanning electron and confocal fluorescence microscopic analyses, which showed that the bacterial cells adhered to and invaded human erythrocytes. In addition, S. pneumoniae cells were found associated with human erythrocytes in cultures of blood from patients with an invasive pneumococcal infection. Erythrocyte invasion assays indicated that LPXTG motif-containing pneumococcal proteins, erythrocyte lipid rafts, and erythrocyte actin remodeling are all involved in the invasion mechanism. In a neutrophil killing assay, the viability of S. pneumoniae co-incubated with erythrocytes was higher than that without erythrocytes. Also, H2O2 killing of S. pneumoniae was nearly completely ineffective in the presence of erythrocytes. These results indicate that even when S. pneumoniae organisms are partially killed by iron ion-induced free radicals, they can still invade erythrocytes. Furthermore, in the presence of erythrocytes, S. pneumoniae can more effectively evade antibiotics, neutrophil phagocytosis, and H2O2 killing. PMID:24194877

  15. The Mechanism of 5-Methyltetrahydrofolate Transport by Human Erythrocytes

    PubMed Central

    Branda, Richard F.; Anthony, Bruce K.; Jacob, Harry S.

    1978-01-01

    The mechanism involved in 5-methyltetrahydrofolate uptake by human cells is poorly understood. To more clearly elucidate this physiologically important process, transport of the vitamin was studied in human erythrocytes. 5-methyltetrahydrofolate uptake was found to increase with reticulocytosis, but measurable incorporation occurred in erythrocyte suspensions depleted of reticulocytes, leukocytes, and platelets, indicating uptake by mature erythrocytes. Incubation of erythrocytes with increasing concentrations of [14C]5-methyltetrahydrofolate resulted in increasing uptake but decreasing percentage incorporation, consistent with saturation of a carrier system. Both influx and efflux phases of uptake were temperature dependent, with almost no transport at 4°C. Uptake of [14C]5-methytetrahydrofolate was effectively inhibited by unlabeled 5-methyltetrahydrofolate, 5-formyltetrahydrofolate, and methotrexate, but not by pteroylglutamic acid. Prior incubation with 5-formyltetrahydrofolate increased uptake of [14C]5-methyltetrahydrofolate, and extracellular 5-formyltetrahydrofolate enhanced efflux of [14C]5-methyltetrahydrofolate. Nearly total depletion of ATP increased uptake of [14C]5-methyltetrahydrofolate, but efflux was unchanged. Column chromatography of membrane-free hemolysate after incubation with [14C]5-methyltetrahydrofolate showed 95% of radioactivity corresponded to marker radioisotope, and no other peak was noted. Thus peripheral erythrocytes incorporate 5-methyltetrahydrofolate by a saturable, temperature-dependent, substrate-specific process which is influenced by counter-transport. This mechanism is qualitatively similar to the carrier-mediated transport of folate compounds previously described in other cell types. Therefore, human erythrocytes should be useful for detailed characterization of this membrane carrier system. PMID:659590

  16. Additional erythrocytic and reticulocytic parameters helpful for diagnosis of hereditary spherocytosis: results of a multicentre study.

    PubMed

    Mullier, François; Lainey, Elodie; Fenneteau, Odile; Da Costa, Lydie; Schillinger, Françoise; Bailly, Nicolas; Cornet, Yvan; Chatelain, Christian; Dogne, Jean-Michel; Chatelain, Bernard

    2011-07-01

    Hereditary spherocytosis (HS) is characterised by weakened vertical linkages between the membrane skeleton and the red blood cell's lipid bilayer, leading to the release of microparticles. All the reference tests suffer from specific limitations. The aim of this study was to develop easy to use diagnostic tool for screening of hereditary spherocytosis based on routinely acquired haematological parameters like percentage of microcytes, percentage of hypochromic cells, reticulocyte counts, and percentage of immature reticulocytes. The levels of haemoglobin, mean cell volume, mean corpuscular haemoglobin concentration, reticulocytes (Ret), immature reticulocytes fraction (IRF), hypochromic erythrocytes (Hypo-He) and microcytic erythrocytes (MicroR) were determined on EDTA samples on Sysmex instruments from a cohort of 45 confirmed SH. The HS group was then compared with haemolytical disorders, microcytic anaemia, healthy individuals and routine samples (n = 1,488). HS is characterised by a high Ret count without an equally elevated IRF. All 45 HS have Ret >80,000/μl and Ret(10(9)/L)/IRF (%) greater than 7.7 (rule 1). Trait and mild HS had a Ret/IRF ratio greater than 19. Moderate and severe HS had increased MicroR and MicroR/Hypo-He (rule 2). Combination of both rules gave predictive positive value and negative predictive value of respectively 75% and 100% (n=1,488), which is much greater than single parameters or existing rules. This simple and fast diagnostic method could be used as an excellent screening tool for HS. It is also valid for mild HS, neonates and ABO incompatibilities and overcomes the lack of sensitivity of electrophoresis in ankyrin deficiencies.

  17. Young Children Counting at Home

    ERIC Educational Resources Information Center

    Griffiths, Rose

    2007-01-01

    Learning to count is something that most children start to do by the time they are about two, and parents know from first-hand experience that family members play a big part in helping with this complex process. In this article, the author describes a project involving families sharing effective counting activities. The project called…

  18. Low White Blood Cell Count

    MedlinePlus

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

  19. Specific erythrocyte binding capacity and biological activity of Plasmodium falciparum erythrocyte binding ligand 1 (EBL-1)-derived peptides

    PubMed Central

    Curtidor, Hernando; Rodríguez, Luis E.; Ocampo, Marisol; López, Ramses; García, Javier E.; Valbuena, John; Vera, Ricardo; Puentes, Álvaro; Vanegas, Magnolia; Patarroyo, Manuel E.

    2005-01-01

    Erythrocyte binding ligand 1 (EBL-1) is a member of the ebl multigene family involved in Plasmodium falciparum invasion of erythrocytes. We found that five EBL-1 high-activity binding peptides (HABPs) bound specifically to erythrocytes: 29895 (41HKKKSGELNNNKSGILRSTY60), 29903 (201LYECGK-KIKEMKWICTDNQF220), 29923 (601CNAILGSYADIGDIVRGLDV620), 29924(621WRDINTNKLSEK-FQKIFMGGY640), and 30018 (2481LEDIINLSKKKKKSINDTSFY2500). We also show that binding was saturable, not sialic acid-dependent, and that all peptides specifically bound to a 36-kDa protein on the erythrocyte membrane. The five HABPs inhibited in vitro merozoite invasion depending on the peptide concentration used, suggesting their possible role in the invasion process. PMID:15659376

  20. Specific erythrocyte binding capacity and biological activity of Plasmodium falciparum erythrocyte binding ligand 1 (EBL-1)-derived peptides.

    PubMed

    Curtidor, Hernando; Rodríguez, Luis E; Ocampo, Marisol; López, Ramses; García, Javier E; Valbuena, John; Vera, Ricardo; Puentes, Alvaro; Vanegas, Magnolia; Patarroyo, Manuel E

    2005-02-01

    Erythrocyte binding ligand 1 (EBL-1) is a member of the ebl multigene family involved in Plasmodium falciparum invasion of erythrocytes. We found that five EBL-1 high-activity binding peptides (HABPs) bound specifically to erythrocytes: 29895 ((41)HKKKSGELNNNKSGILRSTY(60)), 29903 ((201)LYECGK-KIKEMKWICTDNQF(220)), 29923 ((601)CNAILGSYADIGDIVRGLDV(620)), 29924((621)WRDINTNKLSEK-FQKIFMGGY(640)), and 30018 ((2481)LEDIINLSKKKKKSINDTSFY(2500)). We also show that binding was saturable, not sialic acid-dependent, and that all peptides specifically bound to a 36-kDa protein on the erythrocyte membrane. The five HABPs inhibited in vitro merozoite invasion depending on the peptide concentration used, suggesting their possible role in the invasion process.

  1. A 9-yr evaluation of carrier erythrocyte encapsulated adenosine deaminase (ADA) therapy in a patient with adult-type ADA deficiency.

    PubMed

    Bax, Bridget E; Bain, Murray D; Fairbanks, Lynette D; Webster, A David B; Ind, Philip W; Hershfield, Michael S; Chalmers, Ronald A

    2007-10-01

    Adenosine deaminase (ADA) deficiency is an inherited disorder which leads to elevated cellular levels of deoxyadenosine triphosphate (dATP) and systemic accumulation of its precursor, 2-deoxyadenosine. These metabolites impair lymphocyte function, and inactivate S-adenosylhomocysteine hydrolase (SAHH) respectively, leading to severe immunodeficiency. Enzyme replacement therapy with polyethylene glycol-conjugated ADA is available, but its efficacy is reduced by anti-ADA neutralising antibody formation. We report here carrier erythrocyte encapsulated native ADA therapy in an adult-type ADA deficient patient. Encapsulated enzyme is protected from antigenic responses and therapeutic activities are sustained. ADA-loaded autologous carrier erythrocytes were prepared using a hypo-osmotic dialysis procedure. Over a 9-yr period 225 treatment cycles were administered at 2-3 weekly intervals. Therapeutic efficacy was determined by monitoring immunological and metabolic parameters. After 9 yr of therapy, erythrocyte dATP concentration ranged between 24 and 44 micromol/L (diagnosis, 234) and SAHH activity between 1.69 and 2.29 nmol/h/mg haemoglobin (diagnosis, 0.34). Erythrocyte ADA activities were above the reference range of 40-100 nmol/h/mg haemoglobin (0 at diagnosis). Initial increases in absolute lymphocyte counts were not sustained; however, despite subnormal circulating CD20(+) cell numbers, serum immunoglobulin levels were normal. The patient tolerated the treatment well. The frequency of respiratory problems was reduced and the decline in the forced expiratory volume in 1 s and vital capacity reduced compared with the 4 yr preceding carrier erythrocyte therapy. Carrier erythrocyte-ADA therapy in an adult patient with ADA deficiency was shown to be metabolically and clinically effective.

  2. Microwave dielectric measurements of erythrocyte suspensions.

    PubMed Central

    Bao, J Z; Davis, C C; Swicord, M L

    1994-01-01

    Complex dielectric constants of human erythrocyte suspensions over a frequency range from 45 MHz to 26.5 GHz and a temperature range from 5 to 40 degrees C have been determined with the open-ended coaxial probe technique using an automated vector network analyzer (HP 8510). The spectra show two separate major dispersions (beta and gamma) and a much smaller dispersion between them. The two major dispersions are analyzed with a dispersion equation containing two Cole-Cole functions by means of a complex nonlinear least squares technique. The parameters of the equation at different temperatures have been determined. The low frequency behavior of the spectra suggests that the dielectric constant of the cell membrane increases when the temperature is above 35 degrees C. The real part of the dielectric constant at approximately 3.4 GHz remains almost constant when the temperature changes. The dispersion shifts with temperature in the manner of a thermally activated process, and the thermal activation enthalpies for the beta- and gamma-dispersions are 9.87 +/- 0.42 kcal/mol and 4.80 +/- 0.06 kcal/mol, respectively. PMID:8075351

  3. Microwave dielectric measurements of erythrocyte suspensions.

    PubMed

    Bao, J Z; Davis, C C; Swicord, M L

    1994-06-01

    Complex dielectric constants of human erythrocyte suspensions over a frequency range from 45 MHz to 26.5 GHz and a temperature range from 5 to 40 degrees C have been determined with the open-ended coaxial probe technique using an automated vector network analyzer (HP 8510). The spectra show two separate major dispersions (beta and gamma) and a much smaller dispersion between them. The two major dispersions are analyzed with a dispersion equation containing two Cole-Cole functions by means of a complex nonlinear least squares technique. The parameters of the equation at different temperatures have been determined. The low frequency behavior of the spectra suggests that the dielectric constant of the cell membrane increases when the temperature is above 35 degrees C. The real part of the dielectric constant at approximately 3.4 GHz remains almost constant when the temperature changes. The dispersion shifts with temperature in the manner of a thermally activated process, and the thermal activation enthalpies for the beta- and gamma-dispersions are 9.87 +/- 0.42 kcal/mol and 4.80 +/- 0.06 kcal/mol, respectively.

  4. Diabetic Erythrocytes Test by Correlation Coefficient

    PubMed Central

    Korol, A.M; Foresto, P; Darrigo, M; Rosso, O.A

    2008-01-01

    Even when a healthy individual is studied, his/her erythrocytes in capillaries continually change their shape in a synchronized erratic fashion. In this work, the problem of characterizing the cell behavior is studied from the perspective of bounded correlated random walk, based on the assumption that diffractometric data involves both deterministic and stochastic components. The photometric readings are obtained by ektacytometry over several millions of shear elongated cells, using a home-made device called Erythrodeformeter. We have only a scalar signal and no governing equations; therefore the complete behavior has to be reconstructed in an artificial phase space. To analyze dynamics we used the technique of time delay coordinates suggested by Takens, May algorithm, and Fourier transform. The results suggest that on random-walk approach the samples from healthy controls exhibit significant differences from those from diabetic patients and these could allow us to claim that we have linked mathematical nonlinear tools with clinical aspects of diabetic erythrocytes’ rheological properties. PMID:19415139

  5. Free erythrocyte porphyrins in cord blood.

    PubMed

    Gottuso, M A; Oski, B F; Oski, F A

    1978-05-01

    Red cell free erythrocyte porphyrin determinations were performed on cord blood specimens from 236 term infants and on capillary blood specimens from 63 preterm infants weighing less than 1,500 gm, during the first week of life. These results were contrasted with those obtained from 398 normal infants and children ages 1 to 6 years. The mean FEP value for the infants was significantly higher than that observed in the normal control subjects. In 10.5% of the term infants and 15.9% of the preterm infants, values in excess of 120 microgram/dl RBCs, the highest value recorded in the normal subjects, were observed. Elevations in FEP values were not related to either blood lead concentration or hematocrit levels in the infants. Infants with elevated FEP values were found to have lower serum iron and transferrin saturation values than did infants with low FEP values. These findings suggest that elevations in cord blood FEP values may indicate a state of relative iron deficiency present at birth.

  6. Phosphorylation of erythrocyte membrane liberates calcium

    SciTech Connect

    Chauhan, V.P.S.; Brockerhoff, H.

    1986-05-01

    Phosphorylation of permeabilized erythrocyte ghost membranes with ATP results in an increase free calcium level as measured with the help of Ca/sup 2 +/ electrode and /sup 45/Ca. This effect could not be observed in the presence of p/sup -/ chloromercuric benzoate, an inhibitor of kinases. The rise in the free calcium due to phosphorylation of the membrane was accompanied by a decrease in the level of phosphatidylinositol (PI) and an increase in phosphatidylinositolmonophosphate (PIP) and phosphatidylinositolbisphosphate (PIP/sub 2/). These results support the proposal that an inositol shuttle, PI in equilibrium PIP in equilibrium PIP/sub 2/, operates to maintain the intracellular calcium concentration. The cation is believed to be sequestered in a cage formed by the head groups of two acidic phospholipid molecules, e.g., phosphatidylserine and phosphatidylinositol, with the participation of both PO and fatty acid ester CO groups. When the inositol group of such a cage is phosphorylated, inter-headgroup hydrogen bonding between the lipids is broken. As a result the cage opens and calcium is released.

  7. Green hemoprotein of erythrocytes: methemoglobin superoxide transferase

    SciTech Connect

    Kiel, J.L.; McQueen, C.; Erwin, D.N.

    1988-01-01

    Influences of base (pH 10), heat (50 degrees C), microwave radiation (2450 MHz, 103 +/- 4 W/kg), and hydrogen peroxide (5.6 mM) generated by glucose oxidase on oxidation of human oxyhemoglobin to methemoglobin were examined. Conversion of oxyhemoglobin to methemoglobin was followed by the difference in absorbancy of 540 or 542 nm and 576 nm wavelength light versus time. Fresh basic hemolysates auto-oxidized on heating with a zero order rate constant, implying that hemoglobin or another protein saturated with oxyhemoglobin catalyzed the oxidation. Simultaneous microwave irradiation inhibited thermally induced auto-oxidation on the average by 28.6%. However, there was great variability among samples and a decrease in auto-oxidation with aging of individual samples. The auto-oxidation rate was independent of initial oxyhemoglobin concentration. Oxidation of partially purified oxyhemoglobin by hydrogen peroxide was not influenced by microwave irradiation. Adding green hemoprotein isolated from human erythrocytes to the oxyhemoglobin/glucose oxidase reaction mixture yielded absorption spectra (500-600 nm) that were a combination of oxyhemoglobin, deoxyhemoglobin, and methemoglobin spectra. Green hemoprotein was labile in hemolysates but stable in a partially purified ferric form. These results imply that thermally unstable reduced green hemoprotein can reverse oxidation of oxyhemoglobin by hydrogen peroxide and could mediate the thermally induced and microwave inhibited auto-oxidation of oxyhemoglobin.

  8. Evaluation of cell counting and leukocyte differentiation in cerebrospinal fluid controls using hematology analyzers by the German Society for Clinical Chemistry and Laboratory Medicine.

    PubMed

    Kleine, Tilmann O; Nebe, C Thomas; Löwer, Christa; Lehmitz, Reinhard; Geilenkeuser, Wolf-Jochen; Kruse, Rolf; Dorn-Beineke, Alexandra

    2010-06-01

    Manual cell counting in cerebrospinal fluid (CSF) is technique-dependent, time-consuming, and thus costly and prone to inter-operator variability and low precision. Flow cytometry (FCM) with fast hematology analyzers (HAs) appears to improve accuracy and precision of CSF cell analysis; rapid CSF cell analysis is especially needed in emergency laboratories. Ten external trials of the German Society for Clinical Chemistry and Laboratory Medicine evaluated FCM with Coulter (LH750, 755), Abbott CD3200, CD3500, CD3700, CD4000, Sapphire, ADVIA120 CSF assay, and Sysmex XE-2100 single platform analyzers. CSF controls were produced using native blood leukocytes and erythrocytes, resembling CSF and thus rendering the trials feasible and allowing comparison with native manual counting in a Fuchs-Rosenthal chamber and FACScan-CD45-CD14 dual platform analysis, which was used as the reference method. Statistical evaluation was performed using Passing/Bablok regression analysis. Our evaluation revealed significant differences with respect to target values in leukocyte and erythrocyte counts, as well as leukocyte differentiation. These differences were attributed to inaccuracies produced by the HAs, due to blank correction in connection with impedance analysis, leukocyte loss, especially through monocyte injury due to the erythrocyte lysing agent, incomplete erythrocyte lysis, ADVIA cell sphering, cell differentiation using algorithms and peroxidase activity. Erythrocyte counting in the CSF controls was inaccurate with the Coulter and ADVIA analyzers. Evaluation of HAs by means of the CSF controls revealed inaccuracies in cell counting and leukocyte differentiation. Analyzer techniques, used for CSF cell assays, therefore need to be improved.

  9. Increased calcium deposits and decreased Ca2+ -ATPase in erythrocytes of ascitic broiler chickens.

    PubMed

    Li, Kai; Zhao, Lihong; Geng, Guangrui; Ma, Liqin; Dong, Shishan; Xu, Tong; Wang, Jianlin; Wang, Huiyu; Tian, Yong; Qiao, Jian

    2011-06-01

    The decrease of erythrocyte deformability may be one of the predisposing factors for pulmonary hypertension and ascites in broiler chickens. In mammals, the cytoplasmic calcium is a major regulator of erythrocyte deformability. In this study, the erythrocyte deformability was measured, and the precise locations of Ca2+ and Ca2+ -ATPase in the erythrocytes were investigated in chickens with ascites syndrome induced by low ambient temperature. The results showed that ascitic broilers had higher filtration index of erythrocyte compared with control groups, indicating a decrease in erythrocyte deformability in ascitic broilers. The more calcium deposits were observed in the erythrocytes of ascitic broilers compared with those of the age-matched control birds. The Ca2+ -ATPase reactive grains were significantly decreased on the erythrocyte membranes of ascitic broilers. Our data suggest that accumulation of intracellular calcium and inhibition of Ca2+ -ATPase might be important factors for the reduced deformability of the erythrocytes of ascitic broilers.

  10. Uncertainty in measurements by counting

    NASA Astrophysics Data System (ADS)

    Bich, Walter; Pennecchi, Francesca

    2012-02-01

    Counting is at the base of many high-level measurements, such as, for example, frequency measurements. In some instances the measurand itself is a number of events, such as spontaneous decays in activity measurements, or objects, such as colonies of bacteria in microbiology. Countings also play a fundamental role in everyday life. In any case, a counting is a measurement. A measurement result, according to its present definition, as given in the 'International Vocabulary of Metrology—Basic and general concepts and associated terms (VIM)', must include a specification concerning the estimated uncertainty. As concerns measurements by counting, this specification is not easy to encompass in the well-known framework of the 'Guide to the Expression of Uncertainty in Measurement', known as GUM, in which there is no guidance on the topic. Furthermore, the issue of uncertainty in countings has received little or no attention in the literature, so that it is commonly accepted that this category of measurements constitutes an exception in which the concept of uncertainty is not applicable, or, alternatively, that results of measurements by counting have essentially no uncertainty. In this paper we propose a general model for measurements by counting which allows an uncertainty evaluation compliant with the general framework of the GUM.

  11. The origins of counting algorithms.

    PubMed

    Cantlon, Jessica F; Piantadosi, Steven T; Ferrigno, Stephen; Hughes, Kelly D; Barnard, Allison M

    2015-06-01

    Humans' ability to count by verbally labeling discrete quantities is unique in animal cognition. The evolutionary origins of counting algorithms are not understood. We report that nonhuman primates exhibit a cognitive ability that is algorithmically and logically similar to human counting. Monkeys were given the task of choosing between two food caches. First, they saw one cache baited with some number of food items, one item at a time. Then, a second cache was baited with food items, one at a time. At the point when the second set was approximately equal to the first set, the monkeys spontaneously moved to choose the second set even before that cache was completely baited. Using a novel Bayesian analysis, we show that the monkeys used an approximate counting algorithm for comparing quantities in sequence that is incremental, iterative, and condition controlled. This proto-counting algorithm is structurally similar to formal counting in humans and thus may have been an important evolutionary precursor to human counting. © The Author(s) 2015.

  12. Erythrocyte G Protein as a Novel Target for Malarial Chemotherapy

    PubMed Central

    Murphy, Sean C; Harrison, Travis; Hamm, Heidi E; Lomasney, Jon W; Mohandas, Narla; Haldar, Kasturi

    2006-01-01

    Background Malaria remains a serious health problem because resistance develops to all currently used drugs when their parasite targets mutate. Novel antimalarial drug targets are urgently needed to reduce global morbidity and mortality. Our prior results suggested that inhibiting erythrocyte Gs signaling blocked invasion by the human malaria parasite Plasmodium falciparum. Methods and Findings We investigated the erythrocyte guanine nucleotide regulatory protein Gs as a novel antimalarial target. Erythrocyte “ghosts” loaded with a Gs peptide designed to block Gs interaction with its receptors, were blocked in β-adrenergic agonist-induced signaling. This finding directly demonstrates that erythrocyte Gs is functional and that propranolol, an antagonist of G protein–coupled β-adrenergic receptors, dampens Gs activity in erythrocytes. We subsequently used the ghost system to directly link inhibition of host Gs to parasite entry. In addition, we discovered that ghosts loaded with the peptide were inhibited in intracellular parasite maturation. Propranolol also inhibited blood-stage parasite growth, as did other β2-antagonists. β-blocker growth inhibition appeared to be due to delay in the terminal schizont stage. When used in combination with existing antimalarials in cell culture, propranolol reduced the 50% and 90% inhibitory concentrations for existing drugs against P. falciparum by 5- to 10-fold and was also effective in reducing drug dose in animal models of infection. Conclusions Together these data establish that, in addition to invasion, erythrocyte G protein signaling is needed for intracellular parasite proliferation and thus may present a novel antimalarial target. The results provide proof of the concept that erythrocyte Gs antagonism offers a novel strategy to fight infection and that it has potential to be used to develop combination therapies with existing antimalarials. PMID:17194200

  13. An iron stable isotope comparison between human erythrocytes and plasma.

    PubMed

    von Blanckenburg, Friedhelm; Oelze, Marcus; Schmid, Dietmar G; van Zuilen, Kirsten; Gschwind, Hans-Peter; Slade, Alan J; Stitah, Sylvie; Kaufmann, Daniel; Swart, Piet

    2014-11-01

    We present precise iron stable isotope ratios measured by multicollector-ICP mass spectrometry (MC-ICP-MS) of human red blood cells (erythrocytes) and blood plasma from 12 healthy male adults taken during a clinical study. The accurate determination of stable isotope ratios in plasma first required substantial method development work, as minor iron amounts in plasma had to be separated from a large organic matrix prior to mass-spectrometric analysis to avoid spectroscopic interferences and shifts in the mass spectrometer's mass-bias. The (56)Fe/(54)Fe ratio in erythrocytes, expressed as permil difference from the "IRMM-014" iron reference standard (δ(56/54)Fe), ranges from -3.1‰ to -2.2‰, a range typical for male Caucasian adults. The individual subject erythrocyte iron isotope composition can be regarded as uniform over the 21 days investigated, as variations (±0.059 to ±0.15‰) are mostly within the analytical precision of reference materials. In plasma, δ(56/54)Fe values measured in two different laboratories range from -3.0‰ to -2.0‰, and are on average 0.24‰ higher than those in erythrocytes. However, this difference is barely resolvable within one standard deviation of the differences (0.22‰). Taking into account the possible contamination due to hemolysis (iron concentrations are only 0.4 to 2 ppm in plasma compared to approx. 480 ppm in erythrocytes), we model the pure plasma δ(56/54)Fe to be on average 0.4‰ higher than that in erythrocytes. Hence, the plasma iron isotope signature lies between that of the liver and that of erythrocytes. This difference can be explained by redox processes involved during cycling of iron between transferrin and ferritin.

  14. Adenosine signaling in normal and sickle erythrocytes and beyond

    PubMed Central

    Zhang, Yujin; Xia, Yang

    2012-01-01

    Sickle cell disease (SCD) is a debilitating hemolytic genetic disorder with high morbidity and mortality affecting millions of individuals worldwide. Although SCD was discovered more than a century ago, no effective mechanism-based prevention and treatment are available due to poorly understood molecular basis of sickling, the fundamental pathogenic process of the disease. SCD patients constantly face hypoxia. One of the best-known signaling molecules to be induced under hypoxic conditions is adenosine. Recent studies demonstrate that hypoxia-mediated elevated adenosine signaling plays an important role in normal erythrocyte physiology. In contrast, elevated adenosine signaling contributes to sickling and multiple life threatening complications including tissue damage, pulmonary dysfunction and priapism. Here, we summarize recent research on the role of adenosine signaling in normal and sickle erythrocytes, progression of the disease and therapeutic implications. In normal erythrocytes, both genetic and pharmacological studies demonstrate that adenosine can enhance 2,3-bisphosphoglycerate (2,3-BPG) production via A2B receptor (ADORA2B) activation, suggesting that elevated adenosine has an unrecognized role in normal erythrocytes to promote O2 release and prevent acute ischemic tissue injury. However, in sickle erythrocytes, the beneficial role of excessive adenosine-mediated 2,3-BPG induction becomes detrimental by promoting deoxygenation, polymerization of sickle hemoglobin and subsequent sickling. Additionally, adenosine signaling via the A2A receptor (ADORA2A) on invariant natural killer T (iNKT) cells inhibits iNKT cell activation and attenuates pulmonary dysfunction in SCD mice. Finally, elevated adenosine coupled with ADORA2BR activation is responsible for priapism, a dangerous complication seen in SCD. Overall, the research reviewed here reveals a differential role of elevated adenosine in normal erythrocytes, sickle erythrocytes, iNK cells and progression

  15. Cation channels, cell volume and the death of an erythrocyte.

    PubMed

    Lang, Florian; Lang, Karl S; Wieder, Thomas; Myssina, Svetlana; Birka, Christina; Lang, Philipp A; Kaiser, Stephanie; Kempe, Daniela; Duranton, Christophe; Huber, Stephan M

    2003-11-01

    Similar to a variety of nucleated cells, human erythrocytes activate a non-selective cation channel upon osmotic cell shrinkage. Further stimuli of channel activation include oxidative stress, energy depletion and extracellular removal of Cl-. The channel is permeable to Ca2+ and opening of the channel increases cytosolic [Ca2+]. Intriguing evidence points to a role of this channel in the elimination of erythrocytes by apoptosis. Ca2+ entering through the cation channel stimulates a scramblase, leading to breakdown of cell membrane phosphatidylserine asymmetry, and stimulates Ca(2+)-sensitive K+ channels, thus leading to KCl loss and (further) cell shrinkage. The breakdown of phosphatidylserine asymmetry is evidenced by annexin binding, a typical feature of apoptotic cells. The effects of osmotic shock, oxidative stress and energy depletion on annexin binding are mimicked by the Ca2+ ionophore ionomycin (1 microM) and blunted in the nominal absence of extracellular Ca2+. Nevertheless, the residual annexin binding points to additional mechanisms involved in the triggering of the scramblase. The exposure of phosphatidylserine at the extracellular face of the cell membrane stimulates phagocytes to engulf the apoptotic erythrocytes. Thus, sustained activation of the cation channels eventually leads to clearance of affected erythrocytes from peripheral blood. Susceptibility to annexin binding is enhanced in several genetic disorders affecting erythrocyte function, such as thalassaemia, sickle-cell disease and glucose-6-phosphate dehydrogenase deficiency. The enhanced vulnerability presumably contributes to the shortened life span of the affected erythrocytes. Beyond their role in the limitation of erythrocyte survival, cation channels may contribute to the triggering of apoptosis in nucleated cells exposed to osmotic shock and/or oxidative stress.

  16. Mice can count and optimize count-based decisions.

    PubMed

    Çavdaroğlu, Bilgehan; Balcı, Fuat

    2016-06-01

    Previous studies showed that rats and pigeons can count their responses, and the resultant count-based judgments exhibit the scalar property (also known as Weber's Law), a psychophysical property that also characterizes interval-timing behavior. Animals were found to take a nearly normative account of these well-established endogenous uncertainty characteristics in their time-based decision-making. On the other hand, no study has yet tested the implications of scalar property of numerosity representations for reward-rate maximization in count-based decision-making. The current study tested mice on a task that required them to press one lever for a minimum number of times before pressing the second lever to collect the armed reward (fixed consecutive number schedule, FCN). Fewer than necessary number of responses reset the response count without reinforcement, whereas emitting responses at least for the minimum number of times reset the response counter with reinforcement. Each mouse was tested with three different FCN schedules (FCN10, FCN20, FCN40). The number of responses emitted on the first lever before pressing the second lever constituted the main unit of analysis. Our findings for the first time showed that mice count their responses with scalar property. We then defined the reward-rate maximizing numerical decision strategies in this task based on the subject-based estimates of the endogenous counting uncertainty. Our results showed that mice learn to maximize the reward-rate by incorporating the uncertainty in their numerosity judgments into their count-based decisions. Our findings extend the scope of optimal temporal risk-assessment to the domain of count-based decision-making.

  17. White blood cell counting system

    NASA Technical Reports Server (NTRS)

    1972-01-01

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

  18. Genetic control of glycolysis in human erythrocytes

    SciTech Connect

    Gilroy, T.E.; Brewer, G.J.; Sing, C.F.

    1980-03-01

    We have studied heritability of the concentration of each glycolytic intermediate and adenine nucleotide in the cytosol of human erythrocytes obtained from a random sample of apparently healthy young individuals. Preliminary to analysis of heritability, each trait was statistically described and the effects attributable to variation in measured concomitants were removed by regression. Heritability was estimated using the family-set method. This method removes covariances between the index case, sibling and first cousin, due to those environmental determinants of the phenotypic values that are shared with a matched, unrelated control member of the family set. It also removes covariances due to environments that are shared by siblings and first cousins. Heritability was estimated by employing the fact that the variance of differences between first cousins minus the variance of differences between full siblings estimates three-fourths of the additive genetic variance. The heritability estimates for G6P, F6P, ATP and some other metabolite concentrations are high and significantly greater than zero. The heritabilities of G6P and F6P are likely attributable to genetic variation in the in vivo activity of HK and/or PFK, because the concentrations of these metabolites are tightly controlled by the two regulatory enzymes. Statistically significant heritability estimates for HK and PFK mass action ratios stronglysuggest genes are responsible for a portion of the quantitative variation in these enzyme activities. Since HK and PFK regulate glycolysis and the production of ATP, genetic variation in their activities might be causally related to the heritability of ATP concentration.

  19. Prevalence and Diagnosis of Hemotrophic Mycoplasma Infection in Research Sheep and Its Effects on Hematology Variables and Erythrocyte Membrane Fragility

    PubMed Central

    Hampel, Joseph A; Spath, Samantha N; Bergin, Ingrid L; Lim, Ailam; Bolin, Steven R; Dyson, Melissa C

    2014-01-01

    Hemotrophic mycoplasma (hemoplasma) infection in research sheep can confound experimental results and contribute to morbidity and mortality. Prevalence and clinicopathologic studies historically relied on blood-smear diagnosis, but systematic studies using current molecular techniques are warranted. Here we sought to report the prevalence of subclinical infection in our study population, compare diagnostic sensitivity and specificity between blood smears and a PCR assay, and determine the effects of infection on CBC variables and erythrocyte membrane fragility. We collected whole-blood samples from 111 convenience-sampled research sheep. All samples were tested for hemoplasmas by using a PCR assay, blood smears were evaluated for visual presence of hemoplasmas, and CBC and osmotic fragility assays were performed. Subclinical prevalence, according to PCR diagnosis, was 14.1% (14 of 99) in our study population. Relative to the PCR assay, blood-smear diagnosis was 8.3% sensitive and 100% specific for hemoplasma detection. Subclinical infection was associated with changes in MCV, MCHC, RBC distribution width, and absolute monocyte count. Acute infection was associated with changes in RBC mass, Hgb concentration, MCV, MCH, MCHC, and absolute lymphocyte and monocyte counts. Acute infection was associated with increased mean erythrocyte fragility compared with that in uninfected control and treated sheep. We demonstrated that hemoplasma infection is common in our study population, blood-smear evaluation is insensitive at detecting infection, and infection is associated with changes in CBC variables and increased erythrocyte membrane fragility. These findings raise concerns regarding the suitability of hemoplasma-infected sheep for biomedical research. PMID:25527029

  20. [Comparison of photodynamic effect with respect to human and rabbit erythrocytes].

    PubMed

    Galebskaia, L V; Solovtsova, I L; Solov'eva, M A; Zammoeva, D B; Kuz'menkov, A N

    2011-01-01

    Parameters of photoinduced lysis are studied for human and rabbit erythrocytes (photosensibilizer--Radachlorin, the light source--Shuttle HeNe lazer, lambda = 633 nm). The higher sensitivity to irradiation is revealed for rabbit erythrocytes. Treatment of erythrocytes with trypsin showed the surface proteins in human cells to produce a protective effect. Trypsynization of rabbit erythrocytes produced the opposite action--the rate of photohemolysis increased. Results of the study indicate the differences in sensitivity to the photoinduced lysis of erythrocytes of different species and participation of erythrocytes proteins in the effect of photohemolysis.

  1. Counting Triangles to Sum Squares

    ERIC Educational Resources Information Center

    DeMaio, Joe

    2012-01-01

    Counting complete subgraphs of three vertices in complete graphs, yields combinatorial arguments for identities for sums of squares of integers, odd integers, even integers and sums of the triangular numbers.

  2. Oxidative Hemolysis of Erythrocytes Induced by Various Vitamins

    PubMed Central

    Ibrahim, I. H.; Sallam, S. M.; Omar, H.; Rizk, M.

    2006-01-01

    Hemolytic effect of some water-soluble vitamins (niacin B5, pyridoxine B6, thiamine B1 and ascorbic and acid C) on erythrocytes was studied spectrophotometrically at relatively high concentration. The oxidation mechanism of hemoglobin was the same for the used vitamins. Vitamin C was the strongest hemolytic agent in comparison with the other vitamins, while vitamin B1 is the weakest one. The results were confirmed by studying the variation in conductivity of erythrocytes with temperature in the range 20-40°C for the used vitamins at a concentration of 2 mM and after two hours from adding each vitamin to the erythrocytes suspension. The conductivity measurements show that the conductivity for the used vitamins is lower than that for control (without adding vitamin) due to hemoglobin oxidation, also may be due to the electrical reorganization of the erythrocyte membrane after the interaction of the used vitamin with it. The obtained results insure the oxidizing effect of the used vitamins on hemoglobin and consequently their hemolytic effect on erythrocytes. PMID:23674994

  3. Disorders of erythrocyte structure and function in hypertensive patients

    PubMed Central

    Pytel, Edyta; Duchnowicz, Piotr; Jackowska, Paulina; Wojdan, Katarzyna; Koter-Michalak, Maria; Broncel, Marlena

    2012-01-01

    Summary Background The prevalence of hypertension is growing at an alarming rate. Increasing attention is being focussed on the oxidative stress accompanying this disease. In this study we examined the impact of this disease on some parameters of erythrocytes and human blood plasma. Material/Methods We examined the impact of hypertension on some parameters of erythrocytes and human plasma. The study involved 13 patients with hypertension and 19 healthy subjects. We determined lipid peroxidation, SH groups concentration, antioxidants enzymes activity, ATPase activity, total antioxidant capacity, total cholesterol level and erythrocyte membrane fluidity. Results We found an increased level of lipid peroxidation and the concentration of SH groups in membrane proteins in patients with hypertension, and a decrease in the activity of catalase and superoxide dysmutase. No changes were observed in glutathione peroxidase and ATPase activity, level of total antioxidant capacity, total cholesterol level and fluidity of erythrocyte membranes. Conclusions These results suggest the existence of an impaired oxidative balance in hypertensive human erythrocytes. PMID:22847194

  4. Origins and function of 3-ribosylurate in bovid erythrocytes.

    PubMed

    Davids, V; Blackhurst, D M; Katz, A A; Harley, E H

    2012-06-01

    3-Ribosylurate is a dominant feature on high performance liquid chromatography (HPLC) profiles of acid extracts of erythrocytes from cows and buffalo, but is HPLC-undetectable in acid extracts of erythrocytes from all other species examined to date. Various aspects of this unique low molecular weight substance remain unexplored since it was first identified. In this study, the mutation(s) responsible for the appearance of ribosylurate in these cells is shown to be specific to members of both tribes of the Bovinae subfamily (Bovidae family), being detectable in the erythrocytes of both the cow and the buffalo (Bovini tribe) as well as in the kudu (Strepsicerotini tribe), but not in representative species from the other subfamilies of the Bovidae family. More specifically, expression of the mutation(s) seems to be restricted to the erythrocyte lineage of these species, ribosylurate being undetectable in cow white blood cells and primary cultures of fibroblasts. Novel evidence is presented that ribosylurate has antioxidant activity. Accumulation of high levels specifically within the haemoglobin-rich milieu of circulating erythrocytes may serve to protect perfused tissues by removing pathophysiological levels of hydrogen peroxide from plasma. Maintenance of ribosylurate levels may be important in conditions associated with oxidative stress in Bovinae. Copyright © 2011 Elsevier Ltd. All rights reserved.

  5. Very Deep Convolutional Neural Networks for Morphologic Classification of Erythrocytes.

    PubMed

    Durant, Thomas J S; Olson, Eben M; Schulz, Wade L; Torres, Richard

    2017-09-06

    Morphologic profiling of the erythrocyte population is a widely used and clinically valuable diagnostic modality, but one that relies on a slow manual process associated with significant labor cost and limited reproducibility. Automated profiling of erythrocytes from digital images by capable machine learning approaches would augment the throughput and value of morphologic analysis. To this end, we sought to evaluate the performance of leading implementation strategies for convolutional neural networks (CNNs) when applied to classification of erythrocytes based on morphology. Erythrocytes were manually classified into 1 of 10 classes using a custom-developed Web application. Using recent literature to guide architectural considerations for neural network design, we implemented a "very deep" CNN, consisting of >150 layers, with dense shortcut connections. The final database comprised 3737 labeled cells. Ensemble model predictions on unseen data demonstrated a harmonic mean of recall and precision metrics of 92.70% and 89.39%, respectively. Of the 748 cells in the test set, 23 misclassification errors were made, with a correct classification frequency of 90.60%, represented as a harmonic mean across the 10 morphologic classes. These findings indicate that erythrocyte morphology profiles could be measured with a high degree of accuracy with "very deep" CNNs. Further, these data support future efforts to expand classes and optimize practical performance in a clinical environment as a prelude to full implementation as a clinical tool. © 2017 American Association for Clinical Chemistry.

  6. Effects of gemfibrozil on the oxygen transport properties of erythrocytes.

    PubMed Central

    Scatena, R; Nocca, G; Messana, I; De Sole, P; Baroni, S; Zuppi, C; Castagnola, M; Giardina, B

    1995-01-01

    1. In the present study we have investigated the effects of the relatively low plasma concentrations of gemfibrozil (GFZ) found in clinical practice on the oxygen dissociation curve (ODC) of erythrocytes. 2. ODCs were measured at 30 degrees C and 37 degrees C and at pH 7.4: a) both on HbA solution and erythrocytes incubated in vitro with gemfibrozil and clofibric acid; b) on erythrocytes from healthy volunteers treated with a single oral dose of gemfibrozil. 3. These experiments showed a significant drug-induced shift of the ODC towards lower O2 affinity values without any significant modification of metabolic parameters of erythrocytes such as intracellular pH and intraerythrocytic levels of ATP and DPG. 4. In our experimental conditions gemfibrozil appears to lower both in vitro and in vivo, the partial pressure of oxygen required to give 50% of the haemes saturated with oxygen (P50) of erythrocytes from the control value of 24 +/- 0.5 mm Hg to 29 +/- 0.5 mm Hg (mean +/- s.d.; P < 0.02 by ANOVA). 5. These data clearly indicate that therapeutic doses of gemfibrozil may influence the oxygen transport properties of red cells. This effect could have relevant pharmacological and toxicological implications. PMID:7756095

  7. Atomic force microscopy of asymmetric membranes from turtle erythrocytes.

    PubMed

    Tian, Yongmei; Cai, Mingjun; Xu, Haijiao; Ding, Bohua; Hao, Xian; Jiang, Junguang; Sun, Yingchun; Wang, Hongda

    2014-08-01

    The cell membrane provides critical cellular functions that rely on its elaborate structure and organization. The structure of turtle membranes is an important part of an ongoing study of erythrocyte membranes. Using a combination of atomic force microscopy and single-molecule force spectroscopy, we characterized the turtle erythrocyte membrane structure with molecular resolution in a quasi-native state. High-resolution images both leaflets of turtle erythrocyte membranes revealed a smooth outer membrane leaflet and a protein covered inner membrane leaflet. This asymmetry was verified by single-molecule force spectroscopy, which detects numerous exposed amino groups of membrane proteins in the inner membrane leaflet but much fewer in the outer leaflet. The asymmetric membrane structure of turtle erythrocytes is consistent with the semi-mosaic model of human, chicken and fish erythrocyte membrane structure, making the semi-mosaic model more widely applicable. From the perspective of biological evolution, this result may support the universality of the semi-mosaic model.

  8. Integral membrane protein interaction with Triton cytoskeletons of erythrocytes.

    PubMed

    Sheetz, M P

    1979-10-19

    The organization of erythrocyte membrane lipids and proteins has been studied following the release of cytoplasmic components with the non-ionic detergent Triton X-100. After detergent extraction, a detergent-resistant complex called the erythrocyte cytoskeleton is separated from detergent, solubilized lipid and protein by sucrose buoyant density sedimentation. In cytoskeletons prepared under isotonic conditions all of the major erythrocyte membrane proteins are retained except for the integral protein, glycophorin, which is quantitatively solubilized and another integral glycoprotein, band 3, which is only 60% removed. When cytoskeletons are prepared in hypertonic KCl solutions, band 3 is fully solubilized along with bands 2.1 and 4.2 and several minor components. The resulting cytoskeletons have the same morphology as those prepared in isotonic buffer but they are composed of only three major peripheral proteins, spectrin, actin and band 4.1. We have designated this peripheral protein complex the 'shell' of the erythrocyte membrane, and have shown that the attachment of band 3 to the shell satisfies the criteria for a specific interaction. Although Triton did affect erythrocyte shape, cytoskeleton lipid content and the activity of membrane proteases, there was no indication that Triton altered the attachment of band 3 to the shell. We suggest that band 3 attaches to the shell as part of a ternary complex of bands 2.1, 3 and 4.2.

  9. Retention of radiolead by human erythrocytes in vitro

    SciTech Connect

    Barton, J.C.

    1989-06-15

    An in vitro method was developed to assess human erythrocyte lead uptake and release directly, rapidly, and reproducibly; the technique requires small aliquots of blood and uses silicone fluid to separate erythrocytes from their suspending media. Uptake occurred rapidly and was directly related to temperature. Increasing quantities of available elemental lead were associated with increasing absolute quantities but decreasing percentages of uptake. Low values of pH diminished the uptake and enhanced the release of radiolead by erythrocytes, and could be correlated with diminished lead-hemoglobin binding para-Chloromecuribenzoate increased and dithiothreitol inhibited radiolead uptake but neither compound affected lead release, suggesting that sulfhydryl groups are important for lead binding to the erythrocyte. Cyanamide and N-ethylmaleimide did not significantly affect the net uptake or release of radiolead. Calcium disodium EDTA, penicillamine, and dimercaprol significantly reduced lead uptake, although only incubation with dimercaprol resulted in a net removal of lead from erythrocytes. Iron and ceruloplasmin significantly decreased radiolead uptake, but inorganic metal cations other than iron, hyperosmolarity, human serum albumin, cholesterol, and transferrin had no significant effect on uptake or release.

  10. Dielectrophoretic characterization of erythrocytes: positive ABO blood types.

    PubMed

    Srivastava, Soumya K; Daggolu, Prashant R; Burgess, Shane C; Minerick, Adrienne R

    2008-12-01

    Dielectrophoretic manipulation of erythrocytes/red blood cells is investigated as a tool to identify blood type for medical diagnostic applications. Positive blood types of the ABO typing system (A+, B+, AB+ and O+) were tested and cell responses quantified. The dielectrophoretic response of each blood type was observed in a platinum electrode microdevice, delivering a field of 0.025V(pp)/microm at 1 MHz. Responses were recorded via video microscopy for 120 s and erythrocyte positions were tabulated at 20-30 s intervals. Both vertical and horizontal motions of erythrocytes were quantified via image object recognition, object tracking in MATLAB, binning into appropriate electric field contoured regions (wedges) and statistical analysis. Cells of O+ type showed relatively attenuated response to the dielectrophoretic field and were distinguished with greater than 95% confidence from all the other three blood types. AB+ cell responses differed from A+ and B+ blood types likely because AB+ erythrocytes express both the A and B glycoforms on their membrane. This research suggests that dielectrophoresis of untreated erythrocytes beyond simple dilution depends on blood type and could be used in portable blood typing devices.

  11. Calcium imaging of individual erythrocytes: problems and approaches.

    PubMed

    Kaestner, Lars; Tabellion, Wiebke; Weiss, Erwin; Bernhardt, Ingolf; Lipp, Peter

    2006-01-01

    Although in erythrocytes calcium is thought to be important in homeostasis, measurements of this ion concentration are generally seen as rather problematic because of the auto-fluorescence or absorption properties of the intracellular milieu. Here, we describe experiments to assess the usability of popular calcium indicators such as Fura-2, Indo-1 and Fluo-4. In our experiments, Fluo-4 turned out to be the preferable indicator because (i) its excitation and emission properties were least influenced by haemoglobin and (ii) it was the only dye for which excitation light did not lead to significant auto-fluorescence of the erythrocytes. From these results, we conclude that the use of indicators such as Fura-2 together with red blood cells has to be revisited critically. We thus utilized Fluo-4 in erythrocytes to demonstrate a robust but heterogeneous calcium increase in these cells upon stimulation by prostaglandin E(2) and lysophosphatidic acid. For the latter stimulus, we recorded emission spectra of individual erythrocytes to confirm largely unaltered Fluo-4 emission. Our results emphasize that in erythrocytes measurements of intracellular calcium are reliably possible with Fluo-4 and that other indicators, especially those requiring UV-excitation, appear less favourable.

  12. Thallium and rubidium permeability of human and rat erythrocyte membrane.

    PubMed

    Skulskii, I A; Manninen, V; Glasunov, V V

    1990-02-01

    Transport of Tl+ and Rb+ in human and rat erythrocytes was investigated in the presence of ouabain. The chloride-dependent cotransport of Tl+, Rb+ and Na+ was precluded by replacement of Cl- by NO3-. The inward and outward rate constants for the residual fluxes of the cations were determined by measuring the transport of 204Tl and 86Rb in double label experiments. The rate of passive transport of Tl+ exceeded that of Rb+ by one-two orders of magnitude in human as well as rat erythrocytes. The membrane barrier which contributes to the maintenance of ion gradients was shown not to be a barrier for Tl+ which easily penetrates the membrane by an unknown mechanism. In rat erythrocytes the barrier for Rb+ was 10-15 times weaker than that in human red blood cells, while the corresponding ratio of rat/human Tl+ permeabilities was about 1.8-2.0. It follows that Tl+ permeability is only slightly affected by factors modifying the permeability to alkali cations. The increase of temperature from 20 degrees to 37 degrees C resulted in a three-fourfold stimulation of the passive transport of Tl+ both in human and rat erythrocytes. The movement of Tl+ and Rb+ through the erythrocyte membrane differed substantially from their diffusion along the excitable membrane channels characterized both by poor Tl+/K+ selectivity and weak temperature dependence.

  13. Chloramines and hypochlorous acid oxidize erythrocyte peroxiredoxin 2.

    PubMed

    Stacey, Melissa M; Peskin, Alexander V; Vissers, Margreet C; Winterbourn, Christine C

    2009-11-15

    Peroxiredoxin 2 (Prx2) is an abundant thiol protein that is readily oxidized in erythrocytes exposed to hydrogen peroxide. We investigated its reactivity in human erythrocytes with hypochlorous acid (HOCl) and chloramines, relevant oxidants in inflammation. Prx2 was oxidized to a disulfide-linked dimer by HOCl, glycine chloramine (GlyCl), and monochloramine (NH(2)Cl) in a dose-dependent manner. In the absence of added glucose, Prx2 and GSH showed similar sensitivities. Second-order rate constants for the reactions of Prx2 with NH(2)Cl and GlyCl were 1.5 x 10(4) and 8 M(-1) s(-1), respectively. The NH(2)Cl value is approximately 10 times higher than that for GSH, whereas Prx2 is approximately 30 times less sensitive than GSH to GlyCl. Thus, the relative sensitivity of Prx2 to GlyCl is greater in the erythrocyte. Oxidation of erythrocyte Prx2 and GSH was less in the presence of glucose, probably because of recycling. High doses of NH(2)Cl resulted in incomplete regeneration of reduced Prx2, suggesting impairment of the recycling mechanism. Our results show that, although HOCl and chloramines are less selective than H(2)O(2), they nevertheless oxidize Prx2. Exposure to these inflammatory oxidants will result in Prx2 oxidation and could compromise the erythrocyte's ability to resist damaging oxidative insult.

  14. Mice Deficient in the Putative Phospholipid Flippase ATP11C Exhibit Altered Erythrocyte Shape, Anemia, and Reduced Erythrocyte Life Span*♦

    PubMed Central

    Yabas, Mehmet; Coupland, Lucy A.; Cromer, Deborah; Winterberg, Markus; Teoh, Narci C.; D'Rozario, James; Kirk, Kiaran; Bröer, Stefan; Parish, Christopher R.; Enders, Anselm

    2014-01-01

    Transmembrane lipid transporters are believed to establish and maintain phospholipid asymmetry in biological membranes; however, little is known about the in vivo function of the specific transporters involved. Here, we report that developing erythrocytes from mice lacking the putative phosphatidylserine flippase ATP11C showed a lower rate of PS translocation in vitro compared with erythrocytes from wild-type littermates. Furthermore, the mutant mice had an elevated percentage of phosphatidylserine-exposing mature erythrocytes in the periphery. Although erythrocyte development in ATP11C-deficient mice was normal, the mature erythrocytes had an abnormal shape (stomatocytosis), and the life span of mature erythrocytes was shortened relative to that in control littermates, resulting in anemia in the mutant mice. Thus, our findings uncover an essential role for ATP11C in erythrocyte morphology and survival and provide a new candidate for the rare inherited blood disorder stomatocytosis with uncompensated anemia. PMID:24898253

  15. Elongation index of erythrocytes, study of activity of chosen erythrocyte enzymes, and the levels of glutathione, malonyldialdehyde in polycythemia vera (PV).

    PubMed

    Dąbrowski, Z; Dybowicz, A J; Marchewka, A; Teległów, A; Skotnicki, A; Zduńczyk, A; Aleksander, P; Filar-Mierzwa, K

    2011-01-01

    The principal aim of the study was to investigate rheological properties of erythrocytes obtained from patients admitted to the clinic, and diagnosed with polycythemia vera. The polycythemia vera diagnosis was based on the WHO criteria for polycythemia vera. Using a laser rheometer SSD Rheometer-Rheodyn, the elongation index of erythrocytes was determined, indicating an increased rigidity of the erythrocytes in this disease compared with the erythrocytes in healthy people. In order to explain (albeit partially) the reason for reduced elasticity, the erythrocytes of patients with polycythemia were studied for the activity of enzymes - glucose-6-phosphate dehydrogenase and acetylcholinesterase membrane enzyme, as well as the levels of glutathione and malonyldialdehyde. The elevated activities of these enzymes, the glutathione level, and elevated ‰ of reticulocytes, indicated an increased pool of juvenile erythrocyte forms; furthermore, the elevated value of malonyldialdehyde may suggest a lipid peroxidative damage in certain pool of the erythrocyte membrane in blood circulation.

  16. Counting on Using a Number Game

    ERIC Educational Resources Information Center

    Betts, Paul

    2015-01-01

    Counting all and counting on are distinct counting strategies that can be used to compute such quantities as the total number of objects in two sets (Wright, Martland, and Stafford 2010). Given five objects and three more objects, for example, children who use counting all to determine quantity will count both collections; that is, they count…

  17. Storage-Induced Changes in Erythrocyte Membrane Proteins Promote Recognition by Autoantibodies

    PubMed Central

    Dinkla, Sip; Novotný, Věra M. J.; Joosten, Irma; Bosman, Giel J. C. G. M.

    2012-01-01

    Physiological erythrocyte removal is associated with a selective increase in expression of neoantigens on erythrocytes and their vesicles, and subsequent autologous antibody binding and phagocytosis. Chronic erythrocyte transfusion often leads to immunization and the formation of alloantibodies and autoantibodies. We investigated whether erythrocyte storage leads to the increased expression of non-physiological antigens. Immunoprecipitations were performed with erythrocytes and vesicles from blood bank erythrocyte concentrates of increasing storage periods, using patient plasma containing erythrocyte autoantibodies. Immunoprecipitate composition was identified using proteomics. Patient plasma antibody binding increased with erythrocyte storage time, while the opposite was observed for healthy volunteer plasma, showing that pathology-associated antigenicity changes during erythrocyte storage. Several membrane proteins were identified as candidate antigens. The protein complexes that were precipitated by the patient antibodies in erythrocytes were different from the ones in the vesicles formed during erythrocyte storage, indicating that the storage-associated vesicles have a different immunization potential. Soluble immune mediators including complement factors were present in the patient plasma immunoprecipitates, but not in the allogeneic control immunoprecipitates. The results support the theory that disturbed erythrocyte aging during storage of erythrocyte concentrates contributes to transfusion-induced alloantibody and autoantibody formation. PMID:22879923

  18. Platelet-independent adhesion of calcium-loaded erythrocytes to von Willebrand factor

    PubMed Central

    Bierings, Ruben; Meems, Henriet; Mul, Frederik P. J.; Geerts, Dirk; Vlaar, Alexander P. J.; Voorberg, Jan; Hordijk, Peter L.

    2017-01-01

    Adhesion of erythrocytes to endothelial cells lining the vascular wall can cause vaso-occlusive events that impair blood flow which in turn may result in ischemia and tissue damage. Adhesion of erythrocytes to vascular endothelial cells has been described in multiple hemolytic disorders, especially in sickle cell disease, but the adhesion of normal erythrocytes to endothelial cells has hardly been described. It was shown that calcium-loaded erythrocytes can adhere to endothelial cells. Because sickle erythrocyte adhesion to ECs can be enhanced by ultra-large von Willebrand factor multimers, we investigated whether calcium loading of erythrocytes could promote binding to endothelial cells via ultra-large von Willebrand factor multimers. We used (immunofluorescent) live-cell imaging of washed erythrocytes perfused over primary endothelial cells at venular flow rate. Using this approach, we show that calcium-loaded erythrocytes strongly adhere to histamine-stimulated primary human endothelial cells. This adhesion is mediated by ultra-large von Willebrand factor multimers. Von Willebrand factor knockdown or ADAMTS13 cleavage abolished the binding of erythrocytes to activated endothelial cells under flow. Platelet depletion did not interfere with erythrocyte binding to von Willebrand factor. Our results reveal platelet-independent adhesion of calcium-loaded erythrocytes to endothelium-derived von Willebrand factor. Erythrocyte adhesion to von Willebrand factor may be particularly relevant for venous thrombosis, which is characterized by the formation of erythrocyte-rich thrombi. PMID:28249049

  19. Altered Membrane Structure and Surface Potential in Homozygous Hemoglobin C Erythrocytes

    PubMed Central

    Tokumasu, Fuyuki; Nardone, Glenn A.; Ostera, Graciela R.; Fairhurst, Rick M.; Beaudry, Steven D.; Hayakawa, Eri; Dvorak, James A.

    2009-01-01

    Background Hemoglobin C differs from normal hemoglobin A by a glutamate-to-lysine substitution at position 6 of beta globin and is oxidatively unstable. Compared to homozygous AA erythrocytes, homozygous CC erythrocytes contain higher levels of membrane-associated hemichromes and more extensively clustered band 3 proteins. These findings suggest that CC erythrocytes have a different membrane matrix than AA erythrocytes. Methodology and Findings We found that AA and CC erythrocytes differ in their membrane lipid composition, and that a subset of CC erythrocytes expresses increased levels of externalized phosphatidylserine. Detergent membrane analyses for raft marker proteins indicated that CC erythrocyte membranes are more resistant to detergent solubilization. These data suggest that membrane raft organization is modified in CC erythrocytes. In addition, the average zeta potential (a measure of surface electrochemical potential) of CC erythrocytes was ≈2 mV lower than that of AA erythrocytes, indicating that substantial rearrangements occur in the membrane matrix of CC erythrocytes. We were able to recapitulate this low zeta potential phenotype in AA erythrocytes by treating them with NaNO2 to oxidize hemoglobin A molecules and increase levels of membrane-associated hemichromes. Conclusion Our data support the possibility that increased hemichrome deposition and altered lipid composition induce molecular rearrangements in CC erythrocyte membranes, resulting in a unique membrane structure. PMID:19503809

  20. Membrane glycophorins in Sta blood group erythrocytes.

    PubMed

    Blumenfeld, O O; Adamany, A M; Kikuchi, M; Sabo, B; McCreary, J

    1986-04-25

    Structural and immunochemical studies of glycophorins isolated from erythrocytes of an individual homozygous for the M Sta blood group phenotype are described. Reactivities with specific monoclonal antibodies indicated that two major M and N glycophorins were present. The M and N Sta glycophorins were resolved by Lens culinaris lectin affinity chromatography. The N species was not held on the lectin but the M species, like control alpha glycophorins, was retained and could be eluted with alpha-methylmannoside. The two proteins were present in almost equimolar amounts. Studies of the CNBr fragments provided evidence that the structure of M Sta glycophorin is the same as that of the usual M alpha glycophorin but that the N Sta glycophorin is a variant. The amino-terminal octapeptides of the M and N species were similar in amino acid and carbohydrate composition to those isolated, respectively, from M and N alpha glycophorins. The studies focused on CNBr glycopeptide B that, in control alpha glycophorins, extends from amino acid residues 9 to 81. The fragment from the M species exhibited properties identical to those of the corresponding fragment of control alpha glycophorins in terms of size, chromatographic behavior, amino acid and carbohydrate contents and compositions, the presence of O-glycosidically linked saccharides and a single Asn-linked carbohydrate unit. The structures of the O-linked units were inferred experimentally to be NeuAc(alpha 2,3)Gal-(beta 1,3)GalNAc and NeuAc(alpha 2,3)Gal(beta 1,3) [NeuAc(alpha 2,6)]GalNAc, present in a ratio similar to that found in controls; and the Asn-linked unit also appeared to be as in the control. The tryptic glycopeptide pattern of the M Sta glycophorin CNBr fragment B was identical to the pattern of the corresponding control fragment, and the composition of the tryptic peptides suggested sequence identity with the control fragment. In contrast, the N Sta glycophorin yielded two CNBr glycopeptides B; both contained

  1. Determination of somatic mutations in human erythrocytes by cytometry

    SciTech Connect

    Jensen, R.H.; Langlois, R.G.; Bigbee, W.L.

    1985-06-21

    Flow cytometric assays of human erythrocytes labeled with monoclonal antibodies specific for glycophorin A were used to enumerate variant cells that appear in peripheral blood as a result of somatic gene-loss mutations in erythrocyte precursor cells. The assay was performed on erythrocytes from 10 oncology patients who had received at least one treatment from radiation or mutagenic chemotherapy at least 3 weeks before being assayed. The patients were suffering from many different malignancies (e.g., breast, renal, bone, colon and lung), and were treated with several different mutagenic therapeutics (e.g., cisplatinum, adriamycin, daunomycin, or cyclophosphamide). The frequency of these variant cells is an indication of the amount of mutagenic damage accumulated in the individual's erythropoietic cell population. Comparing these results to HPRT clonogenic assays, we find similar baseline frequencies of somatic mutation as well as similar correlation with mutagenic exposures. 9 refs., 3 figs., 1 tab.

  2. The interactions of fibrinogen and dextrans with erythrocytes

    PubMed Central

    Rampling, M.; Sirs, John A.

    1972-01-01

    1. The rate of packing of erythrocytes in whole blood, under a centrifugal field of 200 g, has been studied using an automatic recording centrifuge. 2. Reduction of the supernatant fibrinogen concentration, by repeatedly washing the cells, lowers the rate of packing and reduces the cell flexibility. 3. Resuspending the cells in their own plasma or in isotonic solutions containing fibrinogen restores their flexibility. 4. Rouleaux formation has been shown to have no effect on the rate of packing by comparison of blood diluted with plasma, isotonic NaCl or Ringer—Locke solutions. While the degree of rouleaux formation varied with the diluent used, the rate of packing and packed cell haematocrit were the same, for the same dilution. 5. Both formalin and dextran altered the degree of rouleaux formation and reduced erythrocyte flexibility. Dextran was found to act indirectly on the erythrocyte flexibility by reducing the plasma fibrinogen concentration. PMID:5046146

  3. Optical Assay of Erythrocyte Function in Banked Blood

    NASA Astrophysics Data System (ADS)

    Bhaduri, Basanta; Kandel, Mikhail; Brugnara, Carlo; Tangella, Krishna; Popescu, Gabriel

    2014-09-01

    Stored red blood cells undergo numerous biochemical, structural, and functional changes, commonly referred to as storage lesion. How much these changes impede the ability of erythrocytes to perform their function and, as result, impact clinical outcomes in transfusion patients is unknown. In this study we investigate the effect of the storage on the erythrocyte membrane deformability and morphology. Using optical interferometry we imaged red blood cell (RBC) topography with nanometer sensitivity. Our time-lapse imaging quantifies membrane fluctuations at the nanometer scale, which in turn report on cell stiffness. This property directly impacts the cell's ability to transport oxygen in microvasculature. Interestingly, we found that cells which apparently maintain their normal shape (discocyte) throughout the storage period, stiffen progressively with storage time. By contrast, static parameters, such as mean cell hemoglobin content and morphology do not change during the same period. We propose that our method can be used as an effective assay for monitoring erythrocyte functionality during storage time.

  4. Formulation and Drug Loading Features of Nano-Erythrocytes

    NASA Astrophysics Data System (ADS)

    Dong, Xiaoting; Niu, Yawei; Ding, Yi; Wang, Yuemin; Zhao, Jialan; Leng, Wei; Qin, Linghao

    2017-03-01

    Nano erythrocyte ghosts have recently been used as drug carriers of water-soluble APIs due to inherit biological characteristics of good compatibility, low toxicity, and small side-effect. In this study, we developed a novel drug delivery system based on nano erythrocyte ghosts (STS-Nano-RBCs) to transport Sodium Tanshinone IIA sulfonate (STS) for intravenous use in rat. STS-Nano-RBCs were prepared by hypotonic lysis and by extrusion methods, and its biological properties were investigated compared with STS injection. The results revealed that STS-Nano-RBCs have narrow particle size distribution, good drug loading efficiency, and good stability within 21 days. Compared with STS injection, STS-Nano-RBCs extended the drug release time in vitro and in vivo with better repairing effect on oxidative stress-impaired endothelial cells. These results suggest that the nano erythrocyte ghosts system could be used to deliver STS.

  5. Aggregation of intramembrane particles in erythrocyte membranes treated with diamide.

    PubMed

    Kurantsin-Mills, J; Lessin, L S

    1981-02-20

    Treatment of erythrocytes with diamide (diazene dicarboxylic acid bis-(N,N-dimethylamide)) results in oxidation of sulfhydryl groups of the membrane, and cross-linking of membrane proteins into high molecular weight complexes. Concomitant freeze-etching studies show aggregation of intramembrane particles on the protoplasmic fracture face of erythrocyte ghost membranes treated with the oxidant. Furthermore, after a 3 h incubation of erythrocytes with 10 mM diamide at 37 degrees C, cellular energy levels declined to about 70% of control values. The data suggest that disulfide cross-linking of the major membrane proteins releases the apparent physical occlusion of the band 3 proteins within the interstices of the cytoskeletal shell. This results in the translational mobility of band 3 proteins which is reflected ultra-structurally in the freeze-etch images.

  6. Constitutive equations of erythrocyte membrane incorporating evolving preferred configuration.

    PubMed Central

    Tözeren, A; Skalak, R; Fedorciw, B; Sung, K L; Chien, S

    1984-01-01

    The erythrocyte membrane is modeled as a two-dimensional viscoelastic continuum that evolves under the application of stress. The present analysis of the erythrocyte membrane is motivated by the recent development of knowledge about its molecular structure. The constitutive equations proposed in the present analysis explain in a consistent manner the data on both the deformation and recovery phases of the micropipette experiment. The rheological equations of the present study are applied in a later section to the analysis of a plane membrane deformation that is quantitatively similar to the tank-treading motion of the erythrocytes in a shear field. The computations yield useful information on how the membrane viscosity becomes a more dominant feature in tank-treading motion. The material constants appearing in the proposed constitutive equations may be useful indications of the biochemical state of the membrane in health and disease. PMID:6713066

  7. Specific binding of beta-endorphin to normal human erythrocytes

    SciTech Connect

    Chenet, B.; Hollis, V. Jr.; Kang, Y.; Simpkins, C.

    1986-03-05

    Beta-endorphin (BE) exhibits peripheral functions which may not be mediated by interactions with receptors in the brain. Recent studies have demonstrated binding of BE to both opioid and non-opioid receptors on lymphocytes and monocytes. Abood has reported specific binding of /sup 3/H-dihydromorphine in erythrocytes. Using 5 x 10/sup -11/M /sup 125/I-beta-endorphin and 10/sup -5/M unlabeled BE, they have detected 50% specific binding to human erythrocytes. This finding is supported by results from immunoelectron microscopy using rabbit anti-BE antibody and biotinylated secondary antibody with avidin-biotin complexes horseradish peroxidase. Binding is clearly observed and is confined to only one side of the cells. Conclusions: (1) BE binding to human erythrocytes was demonstrated by radioreceptor assay and immunoelectron microscopy, and (2) BE binding sites exist on only one side of the cells.

  8. Influence of glucose solution on the erythrocyte scattering properties

    NASA Astrophysics Data System (ADS)

    Naumenko, Elena K.

    2007-02-01

    The scattering characteristics of erythrocytes (the coefficients of extinction, scattering, absorption and indicatrixes) were calculated with using the theory Mie for spherical homogeneous spherical particles and the theory for two-layered spherical concentric particles. Transmission spectrums were measured with the spectrophotometer Cary500 in the wavelength range 460-860 n m. Specimens of liquid for imbedding of erythrocytes were preparing by mixing blood plasma a nd 50-% glucose solution with the different concentrations. The volume concentrations (hematocrit) of red blood cells (RBC) were maintained to have the same values in all specimens by adding equal volume of whole blood to immersion liquid of equal volumes. It has been shown that, contrary to theretical prediction, transmission is decreasing for all wavelengths with the addition of glucose solution in interval glucose volume concentrations 0.05 - 0.35-0.4. The subsequent increase of the glucose concentration leads to increasing of spectral transmission as a result of erythrocyte hemolysis.

  9. Effect of osmotic pressure to bioimpedance indexes of erythrocyte suspensions

    NASA Astrophysics Data System (ADS)

    Melnikov, A. A.; Nikolaev, D. V.; Malahov, M. V.; Smirnov, A. V.

    2012-12-01

    In the paper we studied effects of osmotic modification of red blood cells on bioimpedance parameters of erythrocyte suspension. The Cole parameters: the extracellular (Re) and intracellular (Ri) fluid resistance, the Alpha parameter, the characteristic frequency (Fchar) and the cell membranes capacitance (Cm) of concentrated erythrocyte suspensions were measured by bioimpedance analyser in the frequency range 5 - 500 kHz. Erythrocytes were incubated in hypo-, hyper- and isoosmotic solutions to achieve changes in cell volume. It was found that Re and Alpha increased in the suspensions with low osmolarity and decreased in the hypertonic suspensions. Ri, Fchar and Cm were higher in the hyperosmotic and were lower in the hypoosmotic suspensions. Correlations of all BIS parameters with MCV were obtained, but multiple regression analysis showed that only Alpha parameter was independently related to MCV (β=0.77, p=0.01). Thus Alpha parameter may be related the mean corpuscular volume of cells.

  10. Updated role of nitric oxide in disorders of erythrocyte function.

    PubMed

    Kahn, Marc J; Maley, Jason H; Lasker, George F; Kadowitz, Philip J

    2013-03-01

    Nitric oxide is a potent vasodilator that plays a critical role in disorders of erythrocyte function. Sickle cell disease, paroxysmal nocturnal hemoglobinuria and banked blood preservation are three conditions where nitric oxide is intimately related to dysfunctional erythrocytes. These conditions are accompanied by hemolysis, thrombosis and vasoocclusion. Our understanding of the interaction between nitric oxide, hemoglobin, and the vasculature is constantly evolving, and by defining this role we can better direct trials aimed at improving the treatments of disorders of erythrocyte function. Here we briefly discuss nitric oxide's interaction with hemoglobin through the hypothesis regarding Snitrosohemoglobin, deoxyhemoglobin, and myoglobin as nitrite reductases. We then review the current understanding of the role of nitric oxide in sickle cell disease, paroxysmal nocturnal hemoglobinuria, and banked blood, and discuss therapeutics in development to target nitric oxide in the treatment of some of these disorders.

  11. New insights into erythrocyte membrane organization and microelasticity.

    PubMed

    Discher, D E

    2000-03-01

    The erythrocyte membrane's ability to withstand the stresses of circulation has its origins in various levels of structural organization. Central to this membrane's structure-function relationships is a quasi-two-dimensional meshwork of spectrin-actin-protein 4.1 that imparts a resilence to the overlying plasma membrane. New insights into the nonlinear microelasticity of this substructure are being provided by experiments that range from elegant atomic force microscopy tests of single spectrin chains to patterned photobleaching of the micropipette-deformed network. Breakthroughs in atomic level structure determinations are further complemented by emerging biophysical studies of transgenically engineered mice lacking specific erythrocyte membrane proteins. Recent theoretical efforts (computational approaches most notably) also have begun to correlate molecular scale aspects of structure with mechanical measures. All of this recent activity in the biophysics of erythrocyte structure-function is certain to challenge and refine some of the most basic tenets in cell membrane structure-function.

  12. Frequency of enzyme deficiency variants in erythrocytes of newborn infants

    SciTech Connect

    Mohrenweiser, H.W.

    1981-08-01

    The frequency of enzyme deficiency variants, defined as alleles whose products are either absent or almost devoid of normal activity in erythrocytes, was determined for nine erythrocyte enzymes in some 675 newborn infants and in approximately 200 adults. Examples of this type of genetic abnormality, which in the homozygous condition are often associated with significant health consequences, were detected for seven of the nine enzymes studied. Fifteen inherited enzyme deficiency variants in 1809 determinations from adults were identified. Seven of the deficiency variants involved triosephosphate isomerase, a frequency of 0.01 in the newborn population. The average frequency of 2.4/1000 is 2 to 3 times the frequency observed for rare electrophoretic variants of erythrocyte enzymes in this same population.

  13. Identification of novel membrane structures in Plasmodium falciparum infected erythrocytes.

    PubMed

    Clavijo, C A; Mora, C A; Winograd, E

    1998-01-01

    Little is known about the molecular mechanisms underlying the release of merozoites from malaria infected erythrocytes. In this study membranous structures present in the culture medium at the time of merozoite release have been characterized. Biochemical and ultrastructural evidence indicate that membranous structures consist of the infected erythrocyte membrane, the parasitophorous vacuolar membrane and a residual body containing electron dense material. These are subcellular compartments expected in a structure that arises as a consequence of merozoite release from the infected cell. Ultrastructural studies show that a novel structure extends from the former parasite compartment to the surface membrane. Since these membrane modifications are detected only after merozoites have been released from the infected erythrocyte, it is proposed that they might play a role in the release of merozoites from the host cell.

  14. Simulation of dielectric spectra of erythrocytes with various shapes

    NASA Astrophysics Data System (ADS)

    Asami, Koji

    2009-07-01

    Dielectric spectra of erythrocyte suspensions were numerically simulated over a frequency range from 1 kHz to 100 MHz to study the effects of erythrocyte shape on the dielectric spectra. First, a biconcave-discoid model for normal erythrocytes or discocytes was compared with an equivalent oblate spheroid model. The two models showed similar dielectric spectra to each other, suggesting that the oblate spheroid model can be approximately used for discocytes. Second, dielectric spectra were simulated for discocytes deformed by osmotic cell swelling. The deformation resulted in the increase in relaxation intensity and the sharpening of spectrum shape. Finally, dielectric spectra were simulated for echinocytes, stomatocytes and sickle cells that are induced by chemical agents and diseases. The dielectric spectra of echinocytes and stomatocytes were similar to each other, being distinguishable from that of discocytes and quite different from that of sickle cells.

  15. Comparison of three optical methods to study erythrocyte aggregation.

    PubMed

    Zhao, H; Wang, X; Stoltz, J F

    1999-01-01

    The aim of this work was to evaluate three optical methods designed to determine erythrocyte aggregation: Erythroaggregometer (EA; Regulest, France), Laser-assisted Optical Rotational Cell Analyzer (LORCA; Mechatronics, Netherlands) and Fully Automatic Erythrocyte Aggregometer (FAEA; Myrenne, GmbH, Germany). Blood samples were taken from fifty donors (26 males and 24 females). The aggregation of normal red blood cell (RBC) and RBCs suspended in three normo- and hyperaggregating suspending media was studied. The results revealed some significant correlations between parameters measured by these instruments, in particular, between the indexes of aggregation of EA and LORCA. Further, RBC aggregation of multiple myeloma patients was also studied and a hyper erythrocyte aggregation state was found by EA and LORCA.

  16. Standardization of 241Am by digital coincidence counting, liquid scintillation counting and defined solid angle counting.

    PubMed

    Balpardo, C; Capoulat, M E; Rodrigues, D; Arenillas, P

    2010-01-01

    The nuclide (241)Am decays by alpha emission to (237)Np. Most of the decays (84.6%) populate the excited level of (237)Np with energy of 59.54 keV. Digital coincidence counting was applied to standardize a solution of (241)Am by alpha-gamma coincidence counting with efficiency extrapolation. Electronic discrimination was implemented with a pressurized proportional counter and the results were compared with two other independent techniques: Liquid scintillation counting using the logical sum of double coincidences in a TDCR array and defined solid angle counting taking into account activity inhomogeneity in the active deposit. The results show consistency between the three methods within a limit of a 0.3%. An ampoule of this solution will be sent to the International Reference System (SIR) during 2009. Uncertainties were analysed and compared in detail for the three applied methods.

  17. Stimulating Effect of Sclareol on Suicidal Death of Human Erythrocytes.

    PubMed

    Signoretto, Elena; Laufer, Stefan A; Lang, Florian

    2016-01-01

    The diterpene alcohol Sclareol has been proposed for the treatment of malignancy. In analogy to apoptosis of nucleated cells, erythrocytes may enter eryptosis, a suicidal cell death characterized by cell shrinkage and cell membrane scrambling with phosphatidylserine translocation to the erythrocyte surface. Cellular mechanisms involved in the triggering of eryptosis include increase of cytosolic Ca2+ activity ([Ca2+]i), oxidative stress, ceramide, p38 kinase and casein kinase 1α. The present study explored, whether Sclareol induces eryptosis and, if so, shed light on the mechanisms involved. Phosphatidylserine abundance at the erythrocyte surface was estimated from annexin-V-binding, cell volume from forward scatter, [Ca2+]i from Fluo3-fluorescence, abundance of reactive oxygen species (ROS) from 2',7'-dichlorodihydrofluorescein diacetate (DCFDA)-dependent fluorescence, and ceramide abundance at the erythrocyte surface utilizing specific antibodies. Hemolysis was estimated from haemoglobin concentration in the supernatant. A 48 hours exposure of human erythrocytes to Sclareol (≥ 50 µM) significantly increased the percentage of annexin-V-binding cells without significantly modifying the average forward scatter, DCF-fluorescence or ceramide abundance. Sclareol (≥ 50 µM) further triggered hemolysis. Sclareol (100 µM) significantly increased Fluo3-fluorescence, but the effect of Sclareol on annexin-V-binding was not significantly blunted by removal of extracellular Ca2+. Instead, the effect of Sclareol on annexin-V-binding was significantly blunted in the presence of p38 kinase inhibitor skepinone (2 µM) and in the presence of casein kinase 1α inhibitor D4476 (10 µM). Sclareol triggers phospholipid scrambling of the erythrocyte cell membrane, an effect in part due to activation of p38 kinase and casein kinase 1α. © 2016 The Author(s) Published by S. Karger AG, Basel.

  18. Triggering of Eryptosis, the Suicidal Erythrocyte Death, by Perifosine.

    PubMed

    Egler, Jasmin; Lang, Florian

    2017-01-01

    The alkylphospholipid perifosine is used for the treatment of malignancy. The substance is effective by triggering suicidal tumor cell death or apoptosis. Side effects of perifosine include anemia. At least in theory, perifosine-induced anemia could result from stimulation of suicidal erythrocyte death or eryptosis. Hallmarks of eryptosis are cell shrinkage and cell membrane scrambling with phosphatidylserine translocation to the erythrocyte surface. Cellular mechanisms participating in the orchestration of eryptosis include increase of cytosolic Ca2+ activity ([Ca2+]i), oxidative stress, increase of ceramide abundance, as well as activation of staurosporine sensitive protein kinase C and/or of SB203580 sensitive p38 kinase. The present study explored, whether perifosine induces eryptosis and, if so, whether its effect involves and/or requires Ca2+ entry, oxidative stress, ceramide and kinase activation. Flow cytometry was employed to quantify phosphatidylserine exposure at the cell surface from annexin-V-binding, cell volume from forward scatter, [Ca2+]i from Fluo3-fluorescence, reactive oxygen species (ROS) abundance from DCFDA dependent fluorescence, and ceramide abundance utilizing specific antibodies. Hemolysis was estimated from hemoglobin concentration in the supernatant. A 24 hours exposure of human erythrocytes to perifosine (2.5 µg/ml) significantly increased the percentage of annexin-V-binding cells, significantly decreased average forward scatter, significantly increased the percentage of shrunken erythrocytes, and significantly decreased the percentage of swollen erythrocytes. Perifosine significantly increased the percentage of hemolytic erythrocytes. Perifosine significantly increased Fluo3-fluorescence, but decreased DCFDA fluorescence and ceramide abundance. The effect of perifosine on annexin-V-binding was significantly blunted by removal of extracellular Ca2+ and by addition of staurosporine (1 µM), but not by addition of SB203580 (2 µ

  19. Accelerated senescence of human erythrocytes cultured with Plasmodium falciparum.

    PubMed

    Omodeo-Salè, Fausta; Motti, Anna; Basilico, Nicoletta; Parapini, Silvia; Olliaro, Piero; Taramelli, Donatella

    2003-07-15

    Red blood cells infected withPlasmodium falciparum(IRBCs) undergo changes primarily in their membrane composition that contribute to malaria pathogenesis. However, all manifestations (eg, anemia) cannot be accounted for by IRBCs alone. Uninfected erythrocytes (URBCs) may play a role, but they have been under-researched. We wanted to document changes in the erythrocyte membrane that could contribute to URBC reduced life span and malaria-associated anemia. Human erythrocytes were cultured withP falciparumand washed at the trophozoite stage. IRBCs and URBCs were separated on Percoll density gradient, thus obtaining erythrocyte fractions of different densities/ages. IRBC- and URBC-purified membranes were analyzed and compared with control normal erythrocytes (NRBCs) of the same age, from the same donor, kept in the same conditions.P falciparumaccelerated aging of both IRBCs and URBCs, causing a significant shift in the cell population toward the denser (old) fraction. Protein, phospholipid, and cholesterol content were reduced in IRBCs and young URBCs. Young and medium uninfected fractions had higher levels of lipid peroxidation and phospholipid saturation (because of the loss of polyunsaturated fatty acids, PUFAs) and lower phosphatidylserine. In IRBCs, thiobarbituric reactive substances (TBARSs) were higher, and PUFAs and phosphatidylserine lower than in NRBCs and URBCs. In comparison, trophozoite membranes had lower phospholipid (particularly sphingomyelin and phosphatidylserine) and cholesterol content and a higher degree of saturation. Parasite-induced peroxidative damage might account for these modifications. In summary, we demonstrated that membrane damage leading to accelerated senescence of both infected and uninfected erythrocytes will likely contribute to malaria anemia.

  20. Erythrocyte Stiffness during Morphological Remodeling Induced by Carbon Ion Radiation

    PubMed Central

    Zhang, Baoping; Liu, Bin; Zhang, Hong; Wang, Jizeng

    2014-01-01

    The adverse effect induced by carbon ion radiation (CIR) is still an unavoidable hazard to the treatment object. Thus, evaluation of its adverse effects on the body is a critical problem with respect to radiation therapy. We aimed to investigate the change between the configuration and mechanical properties of erythrocytes induced by radiation and found differences in both the configuration and the mechanical properties with involving in morphological remodeling process. Syrian hamsters were subjected to whole-body irradiation with carbon ion beams (1, 2, 4, and 6 Gy) or X-rays (2, 4, 6, and 12 Gy) for 3, 14 and 28 days. Erythrocytes in peripheral blood and bone marrow were collected for cytomorphological analysis. The mechanical properties of the erythrocytes were determined using atomic force microscopy, and the expression of the cytoskeletal protein spectrin-α1 was analyzed via western blotting. The results showed that dynamic changes were evident in erythrocytes exposed to different doses of carbon ion beams compared with X-rays and the control (0 Gy). The magnitude of impairment of the cell number and cellular morphology manifested the subtle variation according to the irradiation dose. In particular, the differences in the size, shape and mechanical properties of the erythrocytes were well exhibited. Furthermore, immunoblot data showed that the expression of the cytoskeletal protein spectrin-α1 was changed after irradiation, and there was a common pattern among its substantive characteristics in the irradiated group. Based on these findings, the present study concluded that CIR could induce a change in mechanical properties during morphological remodeling of erythrocytes. According to the unique characteristics of the biomechanical categories, we deduce that changes in cytomorphology and mechanical properties can be measured to evaluate the adverse effects generated by tumor radiotherapy. Additionally, for the first time, the current study provides a new

  1. Purification and sensitivity of Clostridium chauvoei hemolysin to various erythrocytes.

    PubMed

    Mudenda Hang'ombe, Bernard; Kohda, Tomoko; Mukamoto, Masafumi; Kozaki, Shunji

    2006-07-01

    Using ammonium sulphate fractionation, the Clostridium chauvoei hemolysin was purified by cation exchange chromatography and sephacryl S-100 gel filtration. The molecular mass of the hemolysin, determined by SDS-PAGE was found to be approximately 27kDa. The activity of the hemolysin was determined in erythrocytes of various animals, with sensitivities observed in the order of cow, sheep, chicken, rabbit, rat, mouse, dog and horse. Temperature affected the sensitivity of erythrocytes to C. chauvoei hemolysin. These results may reflect distinct characteristics of the hemolytic activity of C. chauvoei hemolysin and that the hemolysin may be pore-forming.

  2. Electrophysiological studies of malaria parasite-infected erythrocytes: Current status

    PubMed Central

    Staines, Henry M.; Alkhalil, Abdulnaser; Allen, Richard J.; De Jonge, Hugo R.; Derbyshire, Elvira; Egée, Stéphane; Ginsburg, Hagai; Hill, David A.; Huber, Stephan M.; Kirk, Kiaran; Lang, Florian; Lisk, Godfrey; Oteng, Eugene; Pillai, Ajay D.; Rayavara, Kempaiah; Rouhani, Sherin; Saliba, Kevin J.; Shen, Crystal; Solomon, Tsione; Thomas, Serge L. Y.; Verloo, Patrick; Desai, Sanjay A.

    2009-01-01

    The altered permeability characteristics of erythrocytes infected with malaria parasites have been a source of interest for over 30 years. Recent electrophysiological studies have provided strong evidence that these changes reflect transmembrane transport through ion channels in the host erythrocyte plasma membrane. However, conflicting results and differing interpretations of the data have led to confusion in this field. In an effort to unravel these issues, the groups involved recently came together for a week of discussion and experimentation. In this article, the various models for altered transport are reviewed, together with the areas of consensus in the field and those that require a better understanding. PMID:17292372

  3. Laser diffractometry of erythrocyte deformation under hypo-osmotic hemolysis

    NASA Astrophysics Data System (ADS)

    Bessmeltsev, Stanislav S.; Lendiaev, Alexander V.; Skvortsova, Yulia A.; Tarlykov, Vladimir A.

    2000-11-01

    The experimental research and theoretical modeling of average radius and refractive index change of spherocytes aggregation of ill and well people under the swelling in solutions of various osmolarity and by taking into account the rigidity of the erythrocyte membrane have been carried out. The spasmodic character of decrease of spherocyte radius has been discovered for the first time. It took place under an NaCl concentration on the order 0.50 - 0.35%. The correlation of the erythrocyte membrane rigidity with a 'jump' value and its position has been demonstrated. The experimental researches of the refractive index have confirmed the spasmodic model of change to the spherocyte radius.

  4. Erythrocyte-derived optical nano-vesicles as theranostic agents

    NASA Astrophysics Data System (ADS)

    Mac, Jenny T.; Nunez, Vicente; Bahmani, Baharak; Guerrero, Yadir; Tang, Jack; Vullev, Valentine I.; Anvari, Bahman

    2015-07-01

    We have engineered nano-vesicles, derived from erythrocytes, which can be doped with various near infrared (NIR) organic chromophores, including the FDA-approved indocyanine green (ICG). We refer to these vesicles as NIR erythrocyte-mimicking transducers (NETS) since in response to NIR photo-excitation they can generate heat or emit fluorescent light. Using biochemical methods based on reduction amination, we have functionalized the surface of NET with antibodies to target specific biomolecules. We present results that demonstrate the effectiveness of NETs in targeted imaging of cancer cells that over-express the human epidermal growth factor receptor-2 (HER2).

  5. Erythrocyte membrane fatty acids in multiple myeloma patients.

    PubMed

    Jurczyszyn, Artur; Czepiel, Jacek; Gdula-Argasińska, Joanna; Czapkiewicz, Anna; Biesiada, Grażyna; Dróżdż, Mirosław; Perucki, William; Castillo, Jorge J

    2014-10-01

    Mounting data show that fatty acids (FA) and fatty acid synthase (FAS) function could be potential targets for multiple myeloma (MM) therapy. Our study aimed at comparing the FA composition of erythrocyte membranes of MM patients and healthy controls. MM patients had higher saturated FA and n-6 polyunsaturated FA (PUFA) and lower monounsaturated, n-3 PUFA and trans-FA indices than controls. The n-3/n-6 PUFA ratio was lower in MM patients and there was distinct clustering of variants of individual FA in MM patients. The FA content of erythrocyte membrane could serve as a diagnostic and/or predictive biomarker in MM.

  6. Hanford whole body counting manual

    SciTech Connect

    Palmer, H.E.; Rieksts, G.A.; Lynch, T.P.

    1990-06-01

    This document describes the Hanford Whole Body Counting Program as it is administered by Pacific Northwest Laboratory (PNL) in support of the US Department of Energy--Richland Operations Office (DOE-RL) and its Hanford contractors. Program services include providing in vivo measurements of internally deposited radioactivity in Hanford employees (or visitors). Specific chapters of this manual deal with the following subjects: program operational charter, authority, administration, and practices, including interpreting applicable DOE Orders, regulations, and guidance into criteria for in vivo measurement frequency, etc., for the plant-wide whole body counting services; state-of-the-art facilities and equipment used to provide the best in vivo measurement results possible for the approximately 11,000 measurements made annually; procedures for performing the various in vivo measurements at the Whole Body Counter (WBC) and related facilities including whole body counts; operation and maintenance of counting equipment, quality assurance provisions of the program, WBC data processing functions, statistical aspects of in vivo measurements, and whole body counting records and associated guidance documents. 16 refs., 48 figs., 22 tabs.

  7. Calpain-1 knockout reveals broad effects on erythrocyte deformability and physiology

    PubMed Central

    Wieschhaus, Adam; Khan, Anwar; Zaidi, Asma; Rogalin, Henry; Hanada, Toshihiko; Liu, Fei; De Franceschi, Lucia; Brugnara, Carlo; Rivera, Alicia; Chishti, Athar H.

    2014-01-01

    Pharmacological inhibitors of cysteine proteases have provided useful insights into the regulation of calpain activity in erythrocytes. However, the precise biological function of calpain activity in erythrocytes remains poorly understood. Erythrocytes express calpain-1, an isoform regulated by calpastatin, the endogenous inhibitor of calpains. In the present study, we investigated the function of calpain-1 in mature erythrocytes using our calpain-1-null [KO (knockout)] mouse model. The calpain-1 gene deletion results in improved erythrocyte deformability without any measurable effect on erythrocyte lifespan in vivo. The calcium-induced sphero-echinocyte shape transition is compromised in the KO erythrocytes. Erythrocyte membrane proteins ankyrin, band 3, protein 4.1R, adducin and dematin are degraded in the calcium-loaded normal erythrocytes but not in the KO erythrocytes. In contrast, the integrity of spectrin and its state of phosphorylation are not affected in the calcium-loaded erythrocytes of either genotype. To assess the functional consequences of attenuated cytoskeletal remodelling in the KO erythrocytes, the activity of major membrane transporters was measured. The activity of the K+–Cl− co-transporter and the Gardos channel was significantly reduced in the KO erythrocytes. Similarly, the basal activity of the calcium pump was reduced in the absence of calmodulin in the KO erythrocyte membrane. Interestingly, the calmodulin-stimulated calcium pump activity was significantly elevated in the KO erythrocytes, implying a wider range of pump regulation by calcium and calmodulin. Taken together, and with the atomic force microscopy of the skeletal network, the results of the present study provide the first evidence for the physiological function of calpain-1 in erythrocytes with therapeutic implications for calcium imbalance pathologies such as sickle cell disease. PMID:22870887

  8. Python erythrocytes are resistant to α-hemolysin from Escherichia coli.

    PubMed

    Larsen, Casper K; Skals, Marianne; Wang, Tobias; Cheema, Muhammad U; Leipziger, Jens; Praetorius, Helle A

    2011-12-01

    α-Hemolysin (HlyA) from Escherichia coli lyses mammalian erythrocytes by creating nonselective cation pores in the membrane. Pore insertion triggers ATP release and subsequent P2X receptor and pannexin channel activation. Blockage of either P2X receptors or pannexin channels reduces HlyA-induced hemolysis. We found that erythrocytes from Python regius and Python molurus are remarkably resistant to HlyA-induced hemolysis compared to human and Trachemys scripta erythrocytes. HlyA concentrations that induced maximal hemolysis of human erythrocytes did not affect python erythrocytes, but increasing the HlyA concentration 40-fold did induce hemolysis. Python erythrocytes were more resistant to osmotic stress than human erythrocytes, but osmotic stress tolerance per se did not confer HlyA resistance. Erythrocytes from T. scripta, which showed higher osmotic resistance than python erythrocytes, were as susceptible to HlyA as human erythrocytes. Therefore, we tested whether python erythrocytes lack the purinergic signalling known to amplify HlyA-induced hemolysis in human erythrocytes. P. regius erythrocytes increased intracellular Ca²⁺ concentration and reduced cell volume when exposed to 3 mM ATP, indicating the presence of a P2X₇-like receptor. In addition, scavenging extracellular ATP or blocking P2 receptors or pannexin channels reduced the HlyA-induced hemolysis. We tested whether the low HlyA sensitivity resulted from low affinity of HlyA to the python erythrocyte membrane. We found comparable incorporation of HlyA into human and python erythrocyte membranes. Taken together, the remarkable HlyA resistance of python erythrocytes was not explained by increased osmotic resistance, lack of purinergic hemolysis amplification, or differences in HlyA affinity.

  9. Calpain-1 knockout reveals broad effects on erythrocyte deformability and physiology.

    PubMed

    Wieschhaus, Adam; Khan, Anwar; Zaidi, Asma; Rogalin, Henry; Hanada, Toshihiko; Liu, Fei; De Franceschi, Lucia; Brugnara, Carlo; Rivera, Alicia; Chishti, Athar H

    2012-11-15

    Pharmacological inhibitors of cysteine proteases have provided useful insights into the regulation of calpain activity in erythrocytes. However, the precise biological function of calpain activity in erythrocytes remains poorly understood. Erythrocytes express calpain-1, an isoform regulated by calpastatin, the endogenous inhibitor of calpains. In the present study, we investigated the function of calpain-1 in mature erythrocytes using our calpain-1-null [KO (knockout)] mouse model. The calpain-1 gene deletion results in improved erythrocyte deformability without any measurable effect on erythrocyte lifespan in vivo. The calcium-induced sphero-echinocyte shape transition is compromised in the KO erythrocytes. Erythrocyte membrane proteins ankyrin, band 3, protein 4.1R, adducin and dematin are degraded in the calcium-loaded normal erythrocytes but not in the KO erythrocytes. In contrast, the integrity of spectrin and its state of phosphorylation are not affected in the calcium-loaded erythrocytes of either genotype. To assess the functional consequences of attenuated cytoskeletal remodelling in the KO erythrocytes, the activity of major membrane transporters was measured. The activity of the K+-Cl- co-transporter and the Gardos channel was significantly reduced in the KO erythrocytes. Similarly, the basal activity of the calcium pump was reduced in the absence of calmodulin in the KO erythrocyte membrane. Interestingly, the calmodulin-stimulated calcium pump activity was significantly elevated in the KO erythrocytes, implying a wider range of pump regulation by calcium and calmodulin. Taken together, and with the atomic force microscopy of the skeletal network, the results of the present study provide the first evidence for the physiological function of calpain-1 in erythrocytes with therapeutic implications for calcium imbalance pathologies such as sickle cell disease.

  10. Dynamic adhesion of eryptotic erythrocytes to immobilized platelets via platelet phosphatidylserine receptors.

    PubMed

    Walker, Britta; Towhid, Syeda T; Schmid, Evi; Hoffmann, Sascha M; Abed, Majed; Münzer, Patrick; Vogel, Sebastian; Neis, Felix; Brucker, Sara; Gawaz, Meinrad; Borst, Oliver; Lang, Florian

    2014-02-01

    Glucose depletion of erythrocytes triggers suicidal erythrocyte death or eryptosis, which leads to cell membrane scrambling with phosphatidylserine exposure at the cell surface. Eryptotic erythrocytes adhere to endothelial cells by a mechanism involving phosphatidylserine at the erythrocyte surface and CXCL16 as well as CD36 at the endothelial cell membrane. Nothing has hitherto been known about an interaction between eryptotic erythrocytes and platelets, the decisive cells in primary hemostasis and major players in thrombotic vascular occlusion. The present study thus explored whether and how glucose-depleted erythrocytes adhere to platelets. To this end, adhesion of phosphatidylserine-exposing erythrocytes to platelets under flow conditions was examined in a flow chamber model at arterial shear rates. Platelets were immobilized on collagen and further stimulated with adenosine diphosphate (ADP, 10 μM) or thrombin (0.1 U/ml). As a result, a 48-h glucose depletion triggered phosphatidylserine translocation to the erythrocyte surface and augmented the adhesion of erythrocytes to immobilized platelets, an effect significantly increased upon platelet stimulation. Adherence of erythrocytes to platelets was blunted by coating of erythrocytic phosphatidylserine with annexin V or by neutralization of platelet phosphatidylserine receptors CXCL16 and CD36 with respective antibodies. In conclusion, glucose-depleted erythrocytes adhere to platelets. The adhesive properties of platelets are augmented by platelet activation. Erythrocyte adhesion to immobilized platelets requires phosphatidylserine at the erythrocyte surface and CXCL16 as well as CD36 expression on platelets. Thus platelet-mediated erythrocyte adhesion may foster thromboocclusive complications in diseases with stimulated phosphatidylserine exposure of erythrocytes.

  11. Kinetics of viral load and erythrocytic inclusion body formation in Pacific herring artificially infected with erythrocytic necrosis virus.

    PubMed

    Glenn, Jolene A; Emmenegger, Eveline J; Grady, Courtney A; Roon, Sean R; Gregg, Jacob L; Conway, Carla M; Winton, James R; Hershberger, Paul K

    2012-09-01

    Viral erythrocytic necrosis (VEN) is a condition that affects marine and anadromous fish species, including herrings and salmonids, in the Atlantic and Pacific oceans. Infection is frequently associated with severe anemia and causes episodic mortality among wild and hatchery fish when accompanied by additional stressors; VEN can be presumptively diagnosed by (1) light microscopic identification of a single characteristic-a round, magenta-colored, 0.8-μm-diameter inclusion body (IB) within the cytoplasm of erythrocytes and their precursors on Giemsa-stained blood films; or (2) observation (via transmission electron microscopy [TEM]) of the causative iridovirus, erythrocytic necrosis virus (ENV), within erythrocytes or their precursors. To better understand the kinetics of VEN, specific-pathogen-free Pacific herring Clupea pallasii were infected with ENV by intraperitoneal injection. At 1, 4, 7, 10, 14, 21, and 28 d postexposure, samples of blood, spleen, and kidney were collected and assessed (1) via light microscopy for the number of intracytoplasmic IBs in blood smears and (2) via TEM for the number of virions within erythrocytes. The mean prevalence of intracytoplasmic IBs in the blood cells increased from 0% at 0-4 d postexposure to 94% at 28 d postexposure. Viral load within circulating red blood cells peaked at 7 d postexposure, fell slightly, and then reached a plateau. However, blood cells observed within the kidney and spleen tissues demonstrated high levels of ENV between 14 and 28 d postexposure. The results indicate that the viral load within erythrocytes does not correlate well with IB prevalence and that the virus can persist in infected fish for more than 28 d.

  12. Janus kinase 3 is expressed in erythrocytes, phosphorylated upon energy depletion and involved in the regulation of suicidal erythrocyte death.

    PubMed

    Bhavsar, Shefalee K; Gu, Shuchen; Bobbala, Diwakar; Lang, Florian

    2011-01-01

    Janus kinase 3, a tyrosine kinase expressed in haematopoetic tissues, plays a decisive role in T-lymphocyte survival. JAK3 deficiency leads to (Severe) Combined Immunodeficiency (SCID) resulting from enhanced lymphocyte apoptosis. JAK3 is activated by phosphorylation. Nothing is known about expression of JAK3 in erythrocytes, which may undergo apoptosis-like cell death (eryptosis) characterized by cell membrane scrambling with phosphatidylserine exposure and cell shrinkage. Triggers of eryptosis include energy depletion. The present study utilized immunohistochemistry and confocal microscopy to test for JAK3 expression and phosphorylation, and FACS analysis to determine phosphatidylserine exposure (annexin binding) and cell volume (forward scatter). As a result, JAK3 was expressed in erythrocytes and phosphorylated following 24h and 48h glucose depletion. Forward scatter was slightly but significantly smaller in erythrocytes from JAK3-deficient mice (jak3(-/-)) than in erythrocytes from wild type mice (jak3(+/+)). Annexin V binding was similarly low in both genotypes. The JAK3 inhibitors WHI-P131/JANEX-1 (4-(4'-Hydroxyphenyl)amino-6,7-dimethoxyquinazoline, 156μM) and WHI-P154 (4-[(3'-Bromo-4'-hydroxyphenyl)amino]-6,7-dimethoxyquinazoline, 11.2μM) did not significantly modify annexin V binding or forward scatter. Glucose depletion increased annexin V binding, an effect significantly blunted in jak3(-/-) erythrocytes and in the presence of the JAK3 inhibitors. The observations disclose a completely novel role of Janus kinase 3, i.e. the triggering of cell membrane scrambling in energy depleted erythrocytes.

  13. Preservation of bilayer structure in human erythrocytes and erythrocyte ghosts after phospholipase treatment. A 31P-NMR study.

    PubMed

    van Meer, G; de Kruijff, B; op den Kamp, J A; van Deenen, L L

    1980-02-15

    1. Fresh human erythrocytes were treated with lytic and non-lytic combinations of phospholipases A2, C and sphingomyelinase. The 31P-NMR spectra of ghosts derived from such erythrocytes show that, in all cases, the residual phospholipids and lysophospholipids remain organized in a bilayer configuration. 2. A bilayer configuration of the (lyso)phospholipids was also observed after treatment of erythrocyte ghosts with various phospholipases even in the case that 98% of the phospholipid was converted into lysophospholipid (72%) and ceramides (26%). 3. A slightly decreased order of the phosphate group of phospholipid molecules, seen as reduced effective chemical shift anisotropy in the 31P-NMR spectra, was found following the formation of diacyglycerols and ceramides in the membrane of intact erythrocytes. Treatment of ghosts always resulted in an extensive decrease in the order of the phosphate groups. 4. The results allow the following conclusions to made: a. Hydrolysis of phospholipids in intact red cells and ghosts does not result in the formation of non-bilayer configuration of residual phospholipids and lysophospholipids. b. Haemolysis, which is obtained by subsequent treatment of intact cells with sphingomyelinase and phospholipase A2, or with phospholipase C, cannot be ascribed to the formation of non-bilayer configuration of phosphate-containing lipids. c. Preservation of bilayer structure, even after hydrolysis of all phospholipid, shows that other membrane constitutents, e.g. cholesterol and/or membrane proteins play an important role in stabilizing the structure of the erythrocyte membrane. d. A major prerequisite for the application of phospholipases in lipid localization studies, the preservation of a bilayer configuration during phospholipid hydrolysis, is met for the erythrocyte membrane.

  14. Kinetics of viral load and erythrocytic inclusion body formation in pacific herring artificially infected with erythrocytic necrosis virus

    USGS Publications Warehouse

    Glenn, Jolene A.; Emmenegger, Eveline J.; Grady, Courtney A.; Roon, Sean R.; Gregg, Jacob L.; Conway, Carla M.; Winton, James R.; Hershberger, Paul K.

    2012-01-01

    Viral erythrocytic necrosis (VEN) is a condition that affects marine and anadromous fish species, including herrings and salmonids, in the Atlantic and Pacific oceans. Infection is frequently associated with severe anemia and causes episodic mortality among wild and hatchery fish when accompanied by additional stressors; VEN can be presumptively diagnosed by (1) light microscopic identification of a single characteristic—a round, magenta-colored, 0.8-μm-diameter inclusion body (IB) within the cytoplasm of erythrocytes and their precursors on Giemsa-stained blood films; or (2) observation (via transmission electron microscopy [TEM]) of the causative iridovirus, erythrocytic necrosis virus (ENV), within erythrocytes or their precursors. To better understand the kinetics of VEN, specific-pathogen-free Pacific herring Clupea pallasii were infected with ENV by intraperitoneal injection. At 1, 4, 7, 10, 14, 21, and 28 d postexposure, samples of blood, spleen, and kidney were collected and assessed (1) via light microscopy for the number of intracytoplasmic IBs in blood smears and (2) via TEM for the number of virions within erythrocytes. The mean prevalence of intracytoplasmic IBs in the blood cells increased from 0% at 0–4 d postexposure to 94% at 28 d postexposure. Viral load within circulating red blood cells peaked at 7 d postexposure, fell slightly, and then reached a plateau. However, blood cells observed within the kidney and spleen tissues demonstrated high levels of ENV between 14 and 28 d postexposure. The results indicate that the viral load within erythrocytes does not correlate well with IB prevalence and that the virus can persist in infected fish for more than 28 d.

  15. LINEAR COUNT-RATE METER

    DOEpatents

    Henry, J.J.

    1961-09-01

    A linear count-rate meter is designed to provide a highly linear output while receiving counting rates from one cycle per second to 100,000 cycles per second. Input pulses enter a linear discriminator and then are fed to a trigger circuit which produces positive pulses of uniform width and amplitude. The trigger circuit is connected to a one-shot multivibrator. The multivibrator output pulses have a selected width. Feedback means are provided for preventing transistor saturation in the multivibrator which improves the rise and decay times of the output pulses. The multivibrator is connected to a diode-switched, constant current metering circuit. A selected constant current is switched to an averaging circuit for each pulse received, and for a time determined by the received pulse width. The average output meter current is proportional to the product of the counting rate, the constant current, and the multivibrator output pulse width.

  16. Photon Counting - One More Time

    NASA Astrophysics Data System (ADS)

    Stanton, Richard H.

    2012-05-01

    Photon counting has been around for more than 60 years, and has been available to amateurs for most of that time. In most cases single photons are detected using photomultiplier tubes, "old technology" that became available after the Second World War. But over the last couple of decades the perfection of CCD devices has given amateurs the ability to perform accurate photometry with modest telescopes. Is there any reason to still count photons? This paper discusses some of the strengths of current photon counting technology, particularly relating to the search for fast optical transients. Technology advances in counters and photomultiplier modules are briefly mentioned. Illustrative data are presented including FFT analysis of bright star photometry and a technique for finding optical pulses in a large file of noisy data. This latter technique is shown to enable the discovery of a possible optical flare on the polar variable AM Her.

  17. Gender differences in the expression of erythrocyte aggregation in relation to B beta-fibrinogen gene polymorphisms in apparently healthy individuals.

    PubMed

    Ben Assayag, Einor; Bova, Irena; Berliner, Shlomo; Peretz, Hava; Usher, Sali; Shapira, Itzhak; Bornstein, Natan M

    2006-03-01

    An increased erythrocyte aggregation (EA) is associated with capillary slow flow, tissue hypoxemia and endothelial dysfunction. Fibrinogen is a major determinant in the formation of aggregated red blood cells. It has been suggested that the B beta-fibrinogen -455 G/A polymorphism is associated with erythrocyte hyperaggregability in men with coronary artery disease. The purpose of this study was to investigate the influence of the beta-fibrinogen -455 G/A polymorphism on erythrocyte aggregation in apparently healthy individuals. Plasma fibrinogen, red blood cell count, serum lipids, erythrocyte sedimentation rate, and the genotype of the B beta-fibrinogen -455 G/A polymorphism were examined in a cohort of 545 apparently healthy individuals and those with atherothrombotic risk factors. A whole blood erythrocyte aggregation test was performed by using a simple slide test and image analysis. In men, EA levels and plasma fibrinogen levels were significantly higher in subjects carrying the -455 A allele compared to subjects with the -455 GG genotype. This association did not exist in women carrying the fibrinogen -455 A allele. The -455 GA/AA men presented significantly higher correlation between the plasma fibrinogen concentrations and EA. This observation raises the prospect of possible change in the functional properties of the -455 GA/AA fibrinogen, enhancing its ability to induce EH. This study suggests that the B beta-fibrinogen -455 A allele is related to EH in men only. Putative mechanism could be hyperfibrinogenemia and a functional change in the fibrinogen molecule that alters its ability to interact with red blood cells and supports the aggregability of these cells.

  18. Kentucky Kids Count 2001 County Data Book: Families Count.

    ERIC Educational Resources Information Center

    Salley, Valerie

    This Kids Count county data book is the eleventh in a series to measure the well-being of Kentucky's children and focuses on the vital role that families play in ensuring their children's success. Included at the beginning of this document is an executive summary of the databook providing an overview of the statewide data for six child and family…

  19. City & Rural KIDS COUNT Data Book. KIDS COUNT Special Report

    ERIC Educational Resources Information Center

    Annie E. Casey Foundation, 2004

    2004-01-01

    This publication provides the objective data needed to track and monitor the well-being of children in different types of American communities. It is part of the ongoing work of the Casey Foundation -- advanced primarily through our KIDS COUNT initiative -- designed to give policymakers data that can help them better understand how conditions of…

  20. Kentucky Kids Count 2001 County Data Book: Families Count.

    ERIC Educational Resources Information Center

    Salley, Valerie

    This Kids Count county data book is the eleventh in a series to measure the well-being of Kentucky's children and focuses on the vital role that families play in ensuring their children's success. Included at the beginning of this document is an executive summary of the databook providing an overview of the statewide data for six child and family…

  1. Effects of microwave resonance therapy on erythrocyte and plasma proteins and lipids in alcoholics.

    PubMed

    Patisheva, E V; Prokopyeva, V D; Bokhan, N A

    2009-07-01

    The content of lipid peroxides and protein carbonyls in erythrocytes and plasma were elevated in patients with alcoholism during abstinence. A course of microwave resonance therapy reduced the level of lipid peroxide in erythrocytes, but not in the plasma, and significantly decreased the content of protein carbonyls in the plasma and erythrocytes.

  2. ABO Blood Groups Influence Macrophage-mediated Phagocytosis of Plasmodium falciparum-infected Erythrocytes

    PubMed Central

    Branch, Donald R.; Hult, Annika K.; Olsson, Martin L.; Liles, W. Conrad; Cserti-Gazdewich, Christine M.; Kain, Kevin C.

    2012-01-01

    Erythrocyte polymorphisms associated with a survival advantage to Plasmodium falciparum infection have undergone positive selection. There is a predominance of blood group O in malaria-endemic regions, and several lines of evidence suggest that ABO blood groups may influence the outcome of P. falciparum infection. Based on the hypothesis that enhanced innate clearance of infected polymorphic erythrocytes is associated with protection from severe malaria, we investigated whether P. falciparum-infected O erythrocytes are more efficiently cleared by macrophages than infected A and B erythrocytes. We show that human macrophages in vitro and mouse monocytes in vivo phagocytose P. falciparum-infected O erythrocytes more avidly than infected A and B erythrocytes and that uptake is associated with increased hemichrome deposition and high molecular weight band 3 aggregates in infected O erythrocytes. Using infected A1, A2, and O erythrocytes, we demonstrate an inverse association of phagocytic capacity with the amount of A antigen on the surface of infected erythrocytes. Finally, we report that enzymatic conversion of B erythrocytes to type as O before infection significantly enhances their uptake by macrophages to observed level comparable to that with infected O wild-type erythrocytes. These data provide the first evidence that ABO blood group antigens influence macrophage clearance of P. falciparum-infected erythrocytes and suggest an additional mechanism by which blood group O may confer resistance to severe malaria. PMID:23071435

  3. Changes in osmotic fragility of nucleated erythrocytes resulting from blood storage.

    PubMed

    Oyewale, J O

    1994-08-01

    The storage of blood for 24 h at 10 degrees C caused significant changes in osmotic fragility of nucleated erythrocytes of pigeons, peafowls, domestic fowls, lizards and toads. Significant decreases in fragility were seen with pigeon and peafowl erythrocytes. However, the osmotic fragility of domestic fowl, lizard and toad erythrocytes increased significantly.

  4. [Physical essence of erythrocytic sedimentation rate in the gravitation field of the earth].

    PubMed

    Cherniĭ, A N

    2009-01-01

    The erythrocytic sedimentation rate method has been long known in medicine and extensively used in laboratory practice in tuberculosis facilities. However, many authors note that the erythrocytic sedimentation rate phenomenon has not clearly understood. By applying the total theory of relativity and quantum mechanics, the author discloses the physical essence of erythrocytic sedimentation in the gravitation field of the Earth.

  5. Experiment study and FEM simulation on erythrocytes under linear stretching of optical micromanipulation

    NASA Astrophysics Data System (ADS)

    Liu, Ying; Song, Huadong; Zhu, Panpan; Lu, Hao; Tang, Qi

    2017-08-01

    The elasticity of erythrocytes is an important criterion to evaluate the quality of blood. This paper presents a novel research on erythrocytes' elasticity with the application of optical tweezers and the finite element method (FEM) during blood storage. In this work, the erythrocytes with different in vitro times were linearly stretched by trapping force using optical tweezers and the time dependent elasticity of erythrocytes was investigated. The experimental results indicate that the membrane shear moduli of erythrocytes increased with the increasing in vitro time, namely the elasticity was decreasing. Simultaneously, an erythrocyte shell model with two parameters (membrane thickness h and membrane shear modulus H) was built to simulate the linear stretching states of erythrocytes by the FEM, and the simulations conform to the results obtained in the experiment. The evolution process was found that the erythrocytes membrane thicknesses were decreasing. The analysis assumes that the partial proteins and lipid bilayer of erythrocyte membrane were decomposed during the in vitro preservation of blood, which results in thin thickness, weak bending resistance, and losing elasticity of erythrocyte membrane. This study implies that the FEM can be employed to investigate the inward mechanical property changes of erythrocyte in different environments, which also can be a guideline for studying the erythrocyte mechanical state suffered from different diseases.

  6. 21 CFR 864.7360 - Erythrocytic glucose-6-phosphate dehydrogenase assay.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 21 Food and Drugs 8 2013-04-01 2013-04-01 false Erythrocytic glucose-6-phosphate dehydrogenase... § 864.7360 Erythrocytic glucose-6-phosphate dehydrogenase assay. (a) Identification. An erythrocytic glucose-6-phosphate dehydrogenase assay is a device used to measure the activity of the enzyme glucose-6...

  7. 21 CFR 864.7360 - Erythrocytic glucose-6-phosphate dehydrogenase assay.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 21 Food and Drugs 8 2012-04-01 2012-04-01 false Erythrocytic glucose-6-phosphate dehydrogenase... § 864.7360 Erythrocytic glucose-6-phosphate dehydrogenase assay. (a) Identification. An erythrocytic glucose-6-phosphate dehydrogenase assay is a device used to measure the activity of the enzyme glucose-6...

  8. 21 CFR 864.7360 - Erythrocytic glucose-6-phosphate dehydrogenase assay.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Erythrocytic glucose-6-phosphate dehydrogenase... § 864.7360 Erythrocytic glucose-6-phosphate dehydrogenase assay. (a) Identification. An erythrocytic glucose-6-phosphate dehydrogenase assay is a device used to measure the activity of the enzyme glucose-6...

  9. 21 CFR 864.7360 - Erythrocytic glucose-6-phosphate dehydrogenase assay.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 21 Food and Drugs 8 2011-04-01 2011-04-01 false Erythrocytic glucose-6-phosphate dehydrogenase... § 864.7360 Erythrocytic glucose-6-phosphate dehydrogenase assay. (a) Identification. An erythrocytic glucose-6-phosphate dehydrogenase assay is a device used to measure the activity of the enzyme glucose-6...

  10. 21 CFR 864.7360 - Erythrocytic glucose-6-phosphate dehydrogenase assay.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 21 Food and Drugs 8 2014-04-01 2014-04-01 false Erythrocytic glucose-6-phosphate dehydrogenase... § 864.7360 Erythrocytic glucose-6-phosphate dehydrogenase assay. (a) Identification. An erythrocytic glucose-6-phosphate dehydrogenase assay is a device used to measure the activity of the enzyme glucose-6...

  11. Counting Multiplicity over Infinite Alphabets

    NASA Astrophysics Data System (ADS)

    Manuel, Amaldev; Ramanujam, R.

    In the theory of automata over infinite alphabets, a central difficulty is that of finding a suitable compromise between expressiveness and algorithmic complexity. We propose an automaton model where we count the multiplicity of data values on an input word. This is particularly useful when such languages represent behaviour of systems with unboundedly many processes, where system states carry such counts as summaries. A typical recognizable language is: “every process does at most k actions labelled a”. We show that emptiness is elementarily decidable, by reduction to the covering problem on Petri nets.

  12. Associations of erythrocyte fatty acid patterns with insulin resistance

    USDA-ARS?s Scientific Manuscript database

    Background: Synergistic and/or additive effects on cardiometabolic risk may be missed by examining individual fatty acids (FA). A pattern analysis may be a more useful approach. As well, it remains unclear whether erythrocyte fatty acid composition relates to insulin resistance among Hispanic/Latino...

  13. 21 CFR 864.6700 - Erythrocyte sedimentation rate test.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Erythrocyte sedimentation rate test. 864.6700 Section 864.6700 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES HEMATOLOGY AND PATHOLOGY DEVICES Manual Hematology Devices § 864.6700...

  14. Phosphorylation of intact erythrocytes in human muscular dystrophy

    SciTech Connect

    Johnson, R.M.; Nigro, M.

    1986-04-01

    The uptake of exogenous /sup 32/Pi into the membrane proteins of intact erythrocytes was measured in 8 patients with Duchenne muscular dystrophy. No abnormalities were noted after autoradiographic analysis. This contrasts with earlier results obtained when isolated membranes were phosphorylated with gamma-(/sup 32/P)ATP, and suggests a possible reinterpretation of those experiments.

  15. MODULATION OF HYPOXIC PULMONARY VASOCONSTRICTION BY ERYTHROCYTIC NITRIC OXIDE

    EPA Science Inventory

    Abstract
    American Heart Association 2001

    Modulation of Hypoxic Pulmonary Vasoconstriction by Erythrocytic NO
    McMahon TJ1, Gow AJ1, Huang YCT4, Stamler JS1,2,3
    Departments of Medicine1 and Biochemistry2, and Howard Hughes Medical Institute3,
    Duke University Med...

  16. 2,3-Diphosphoglycerate and ATP dissociate erythrocyte membrane skeletons.

    PubMed

    Sheetz, M P; Casaly, J

    1980-10-25

    Since ATP and 2,3-diphosphoglycerate cause an increase in the lateral mobility of integral membrane proteins in the erythrocyte (Schindler, M., Koppel, D., and Sheetz, M. P. (1980) Proc. Natl. Acad. Sci. U. S. A. 77, 1457-1461), we have studied their effects on the membrane skeletal complex or shell (composed of spectrin, actin, and bands 4.1 (78,000 daltons) and 4.9 (50,000 daltons)) and its interaction with the erythrocyte membrane. Both phosphate compounds dissociated the delipidated shell complex, with half-maximal dissociation at 2.5 mM 2,3-diphosphoglycerate and 8 mM ATP, whereas equivalent concentrations of EDTA did not. Concomitant with complex dissociation, spectrin was solubilized but band 4.1 and actin remained in a complexed or polymeric form. When proteins which were involved in linking spectrin to the membrane were present on the shell, higher concentrations of the phosphate compounds still dissociated the complex but less spectrin was solubilized. Treatment of erythrocyte membranes with the same phosphate compounds caused membrane vesiculation but no proteins were solubilized. We suggest that ATP and 2,3-diphosphoglycerate, at concentrations which are normally present in erythrocytes, can weaken associations in the shell but will not dissociate the complex from membrane attachment sites.

  17. MODULATION OF HYPOXIC PULMONARY VASOCONSTRICTION BY ERYTHROCYTIC NITRIC OXIDE

    EPA Science Inventory

    Abstract
    American Heart Association 2001

    Modulation of Hypoxic Pulmonary Vasoconstriction by Erythrocytic NO
    McMahon TJ1, Gow AJ1, Huang YCT4, Stamler JS1,2,3
    Departments of Medicine1 and Biochemistry2, and Howard Hughes Medical Institute3,
    Duke University Med...

  18. Erythrocyte aldose reductase activity and sorbitol levels in diabetic retinopathy

    PubMed Central

    Satyanarayana, A.; Balakrishna, N.; Ayyagari, Radha; Padma, M.; Viswanath, K.; Petrash, J. Mark

    2008-01-01

    Purpose Activation of polyol pathway due to increased aldose reductase (ALR2) activity has been implicated in the development of diabetic complications including diabetic retinopathy (DR), a leading cause of blindness. However, the relationship between hyperglycemia-induced activation of polyol pathway in retina and DR is still uncertain. We investigated the relationship between ALR2 levels and human DR by measuring ALR2 activity and its product, sorbitol, in erythrocytes. Methods We enrolled 362 type 2 diabetic subjects (T2D) with and without DR and 66 normal subjects in this clinical case-control study. Clinical evaluation of DR in T2D patients was done by fundus examination. ALR2 activity and sorbitol levels along with glucose and glycosylated hemoglobin (HbA1C) levels in erythrocytes were determined. Results T2D patients with DR showed significantly higher specific activity of ALR2 as compared to T2D patients without DR. Elevated levels of sorbitol in T2D patients with DR, as compared to T2D patients without DR, corroborated the increased ALR2 activity in erythrocytes of DR patients. However, the increased ALR2 activity was not significantly associated with diabetes duration, age, and HbA1C in both the DR group and total T2D subjects. Conclusions Levels of ALR2 activity as well as sorbitol in erythrocytes may have value as a quantitative trait to be included among other markers to establish a risk profile for development of DR. PMID:18385795

  19. The Heme Connection: Linking Erythrocytes and Macrophage Biology

    PubMed Central

    Alam, Md Zahidul; Devalaraja, Samir; Haldar, Malay

    2017-01-01

    Erythroid function and development is intimately linked to macrophages. The primary function of erythrocytes is oxygen delivery, which is mediated by iron-containing hemoglobin. The major source of this iron is a recycling pathway where macrophages scavenge old and damaged erythrocytes to release iron contained within the heme moiety. Macrophages also promote erythropoiesis by providing a supportive niche in the bone marrow as an integral component of “erythorblastic islands.” Importantly, inflammation leads to alterations in iron handling by macrophages with significant impact on iron homeostasis and erythropoiesis. The importance of macrophages in erythropoiesis and iron homeostasis is well established and has been extensively reviewed. However, this developmental relationship is not one way, and erythrocytes can also regulate macrophage development and function. Erythrocyte-derived heme can induce the development of iron-recycling macrophages from monocytes, engage pattern recognition receptors to activate macrophages, and act as ligand for specific nuclear receptors to modulate macrophage function. Here, we discuss the role of heme as a signaling molecule impacting macrophage homeostasis. We will review these actions of heme within the framework of our current understanding of the role of micro-environmental factors in macrophage development and function. PMID:28167947

  20. Sulphurous mineral water oral therapy: effects on erythrocyte metabolism.

    PubMed

    Albertini, Maria Cristina; Teodori, Laura; Accorsi, Augusto; Soukri, Abdelaziz; Campanella, Luigi; Baldoni, Francesco; Dachà, Marina

    2008-10-01

    The ingestion of water containing hydrogen sulphide (H(2)S) is common in spring sulphurous mineral water (SMW) therapy. We hypothesized that observed detrimental effects are related to the alteration of erythrocytes metabolism caused by H(2)S. To verify our hypothesis, we treated 20 healthy volunteers with SMW and evidenced an increase of methemoglobin concentration, an inhibition of both erythrocyte glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and glucose-6-phosphate dehydrogenase (G6PDH) activities. To investigate the mechanism of H(2)S effect on GAPDH activity, an in vitro study was performed by incubating both erythrocytes from 12 healthy volunteers and purified GAPDH with buffered [(35)S]-H(2)S labelled sulphurous water. The interaction between H(2)S and NAD(+)(H), was also investigated. The results indicate that a direct reaction between GAPDH and H(2)S does not occur and the observed decrease of GAPDH activity is to ascribe to the reaction between H(2)S and NAD(+)(H). This may lead to GAPDH inhibition by two ways, namely (i) cellular NAD(+)(H) reduced availability and (ii) catalytic site blockage. In conclusion, our results show that among the detrimental effects of SMW administration are erythrocyte GAPDH and G6PDH activity inhibition and increased methemoglobin concentration. A mechanism to explain the occurrence of these biochemical events is also proposed.

  1. Genotoxic evaluation of pirfenidone using erythrocyte rodent micronucleus assay.

    PubMed

    Alcántar-Díaz, Blanca E; Gómez-Meda, Belinda C; Zúñiga-González, Guillermo M; Zamora-Perez, Ana L; González-Cuevas, Jaime; Alvarez-Rodríguez, Bertha A; Sánchez-Parada, María Guadalupe; García-Bañuelos, Jesús J; Armendáriz-Borunda, Juan

    2012-08-01

    Pirfenidone is a non-steroidal antifibrotic compound that has been proposed in clinical protocols and experimental studies as a pharmacological treatment for fibroproliferative diseases. The objective of this study was to determine the genotoxicity or cytotoxicity of three doses of pirfenidone using the micronuclei test in peripheral blood erythrocytes of rodent models. Pirfenidone was administered orally to Balb-C mice for 3 days, and also was administered topically to hairless Sprague Dawley rats during the final stage of gestation. Mice were sampled every 24 h over the course of 6 days; pregnant rats were sampled every 24 h during the last 6 days of gestation, and pups were sampled at birth. Blood smears were analyzed and the frequencies of micronucleated erythrocytes (MNEs), micronucleated polychromatic erythrocytes (MNPCEs), and the proportion of polychromatic erythrocytes (PCEs), were recorded in samples from mice, pregnant rats and rat neonates. Increases in MN frequencies (p<0.03) were noted only in the positive control groups. No genotoxic effects or decreased PCE values were observed neither in newborn rats transplacentally exposed to pirfenidone, or in two adult rodent models when pirfenidone was administered orally or topically.

  2. Plasma thiamin pyrophosphate and erythrocyte transketolase in chronic alcoholism.

    PubMed

    McLaren, D S; Docherty, M A; Boyd, D H

    1981-06-01

    Thiamin status in patients with an alcohol problem was studied before and after intramuscular thiamin hydrochloride. Results for erythrocyte transketolase activity and plasma thiamin pyrophosphate are compared. Plasma thiamin pyrophosphate values for healthy human subjects are reported for the first time. Advantages of plasma thiamin pyrophosphate in the assessment of thiamin status of patients are discussed.

  3. Changes in erythrocytic deformability and plasma viscosity in neonatal ictericia.

    PubMed

    Bonillo-Perales, A; Muñoz-Hoyos, A; Martínez-Morales, A; Molina-Carballo, A; Uberos-Fernández, J; Puertas-Prieto, A

    1999-01-01

    We studied 45 full-term newborns divided into 3 groups. Group 1: 17 newborns with bilirubin <10 mg/dL; Group 2: 18 newborns with hemolytic ictericia (bilirubin 11-20 mg/dL) and Group 3: 10 newborns with moderate hemolytic ictericia needing exchange transfusion. The following were studied: erythrocytic deformability, plasma viscosity, plasmatic osmolarity, seric bilirubin, bilirubin/albumin ratio, free fatty acids and corpuscular volume of the erythrocytes. In full-term newborns, the following are risk factors for increased erythrocytic rigidity: neonatal hemolytic illness (p = 0.004, odds ratio: 7.02), increases in total bilirubin (p = 0.02, odds ratio: 4.3) and increases in the bilirubin/albumin ratio (p = 0.025, odds ratio: 4.25). Furthermore, the most important risk factor for high plasma viscosity is also neonatal hemolytic illness (p = 0.01, odds ratio: 2.30). The role of total bilirubin is also important (p = 0.09, odds ratio: 2.10), while that of the bilirubin/albumin ratio (p = 0.012, NS) is less so. The greater the hemolysis, the greater the erythrocytic rigidity and plasma viscosity (p < 0.01). In full-term newborns with moderate ictericia, hemolytic illness and increases in the bilirubin/albumin ratio are accompanied by rheological alterations that could affect cerebral microcirculation and cause a neurological deficit not exclusively related to the levels of bilirubin in plasma.

  4. Evaluation of methylglyoxal toxicity in human erythrocytes, leukocytes and platelets.

    PubMed

    Prestes, Alessandro de Souza; Dos Santos, Matheus Mülling; Ecker, Assis; Zanini, Daniela; Schetinger, Maria Rosa Chitolina; Rosemberg, Denis Broock; da Rocha, João Batista Teixeira; Barbosa, Nilda Vargas

    2017-05-01

    Methylglyoxal (MG) is a reactive dicarbonyl metabolite originated mainly from glucose degradation pathway that plays an important role in the pathogenesis of diabetes mellitus (DM). Reactions of MG with biological macromolecules (proteins, DNA and lipids) can induce cytotoxicity and apoptosis. Here, human erythrocytes, leukocytes and platelets were acutely exposed to MG at concentration ranging from 0.025 to 10 mM. Afterwards, hemolysis and osmotic fragility in erythrocytes, DNA damage and cell viability in leukocytes, and the activity of purinergic ecto-nucleotidases in platelets were evaluated. The levels of glycated products from leukocytes and free amino groups from erythrocytes and platelets were also measured. MG caused fragility of membrane, hemolysis and depletion of amino groups in erythrocytes. DNA damage, loss of cell viability and increased levels of glycated products were observed in leukocytes. In platelets, MG inhibited the activity of enzymes NTPDase, 5'-nucleotidase and adenosine deaminase (ADA) without affecting the levels of free amino groups. Our findings provide insights for understanding the mechanisms involved in MG acute toxicity towards distinct blood cells.

  5. Microscopic evaluation of vesicles shed by erythrocytes at elevated temperatures.

    PubMed

    Moore, Timothy; Sorokulova, Iryna; Pustovyy, Oleg; Globa, Ludmila; Pascoe, David; Rudisill, Mary; Vodyanoy, Vitaly

    2013-11-01

    The images of human erythrocytes and vesicles were analyzed by a light microscopy system with spatial resolution of better than 90 nm. The samples were observed in an aqueous environment and required no freezing, dehydration, staining, shadowing, marking, or any other manipulation. Temperature elevation resulted in significant concentration increase of structurally transformed erythrocytes (echinocytes) and vesicles in the blood. The process of vesicle separation from spiculated erythrocytes was video recorded in real time. At a temperature of 37°C, mean vesicle concentrations and diameters were found to be 1.50 ± 0.35 × 10(6) vesicles per microliter and 0.365 ± 0.065 μm, respectively. The vesicle concentration increased approximately threefold as the temperature increased from 37 to 40°C. It was estimated that 80% of all vesicles found in the blood are smaller than 0.4 μm. Accurate account of vesicle numbers and dimensions suggest that 86% of the lost erythrocyte material is lost not by vesiculation but by another, as yet, unknown mechanism. Copyright © 2013 Wiley Periodicals, Inc.

  6. Effect of 8-alkylberberine homologues on erythrocyte membrane.

    PubMed

    Yong, Yang; Ye, Xiao-Li; Zhang, Bao-Shun; Li, Xue-Gang

    2011-05-01

    8-alkylberberine homologues (Ber-C8-n, where n indicates carbon atom number of gaseous normal alkyl at 8 position, n = 0, 2, 4, 6, 8, 10, 12, or 16) were synthesized and their effects on the hemolysis of rabbit erythrocyte, the fluidity of membrane and the fluorescence of membrane protein were investigated by fluorescence analysis technique. Ber-C8-n with mediate length alkyl (4 < n < 10) exhibited obvious hemolysis effect on rabbit erythrocyte when their concentration exceed 1.25 x10(-4) mol/L, and Ber-C8-8 displayed the highest hemolysis effect among all tested homologues. All of Ber-C8-n influenced the fluidity of erythrocyte membrane to different extents, which exhibited an obvious dose-effect relationship. The effect of Ber-C8-n on fluidity increased as the length of alkyl chain was elongated and decreased gradually when the alkyl carbon atoms exceeded 8. The fluorescence of erythrocyte membrane protein was quenched by Ber-C8-n, which showed a similar changing tendency on membrane fluidity. Experiments in vitro suggested that disturbing effects of Ber-C8-n on the conformation and function of membrane protein leaded to the changes of membrane fluidity and stability, and then the membrane was broken down.

  7. Stabilizing factors of phospholipid asymmetry in the erythrocyte membrane.

    PubMed

    Dressler, V; Haest, C W; Plasa, G; Deuticke, B; Erusalimsky, J D

    1984-08-22

    Transbilayer reorientation (flip) of exogenous lysophospholipids and changes of the transbilayer distribution of endogenous phospholipids were studied in human erythrocytes and membrane vesicles. (1) Exogenous lysophosphatidylserine irreversibly accumulates in the inner membrane layer of resealed ghosts of human erythrocytes. (2) This accumulation even occurs after complete loss of asymmetric distribution of endogenous phosphatidylethanolamine and partial loss of phosphatidylserine asymmetry in diamide-treated cells. (3) Formation of inside-out and right-side-out vesicles from erythrocyte membranes results in a loss of endogenous phospholipid asymmetry as well as of the ability to establish asymmetry of exogenous lysophosphatidylserine. Rates of transbilayer reorientation of lysophospholipids for the vesicles, however, are comparable to those for intact cells. (4) Loss of endogenous asymmetry of phosphatidylserine is also observed in vesicles isolated from erythrocytes after heat denaturation of spectrin. The asymmetry in the residual cells is maintained. (5) In contrast to the loss of asymmetry of phosphatidylethanolamine and of phosphatidylserine, the asymmetry of sphingomyelin is completely maintained in the vesicles. (6) The stability of phospholipid asymmetry in the native cell is discussed in terms of a limitation of access of phospholipids to hypothetical reorientation sites. Such a limitation may either be the result of interaction of phospholipids with the membrane skeleton as in case of phosphatidylserine and phosphatidylethanolamine, or the result of lipid-lipid interactions as in case of sphingomyelin.

  8. Erythrocyte antioxidant protection of rose hips (Rosa spp.).

    PubMed

    Widén, C; Ekholm, A; Coleman, M D; Renvert, S; Rumpunen, K

    2012-01-01

    Rose hips are popular in health promoting products as the fruits contain high content of bioactive compounds. The aim of this study was to investigate whether health benefits are attributable to ascorbic acid, phenols, or other rose-hip-derived compounds. Freeze-dried powder of rose hips was preextracted with metaphosphoric acid and the sample was then sequentially eluted on a C(18) column. The degree of amelioration of oxidative damage was determined in an erythrocyte in vitro bioassay by comparing the effects of a reducing agent on erythrocytes alone or on erythrocytes pretreated with berry extracts. The maximum protection against oxidative stress, 59.4 ± 4.0% (mean ± standard deviation), was achieved when incubating the cells with the first eluted meta-phosphoric extract. Removal of ascorbic acid from this extract increased the protection against oxidative stress to 67.9 ± 1.9%. The protection from the 20% and 100% methanol extracts was 20.8 ± 8.2% and 5.0 ± 3.2%, respectively. Antioxidant uptake was confirmed by measurement of catechin by HPLC-ESI-MS in the 20% methanol extract. The fact that all sequentially eluted extracts studied contributed to protective effects on the erythrocytes indicates that rose hips contain a promising level of clinically relevant antioxidant protection.

  9. 40 CFR 799.9539 - TSCA mammalian erythrocyte micronucleus test.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... as sampled in bone marrow and/or peripheral blood cells of animals, usually rodents. (2) The purpose... immature erythrocytes. (N) Number of cells analyzed per animal. (O) Criteria for considering studies as... micronuclei is facilitated in these cells because they lack a main nucleus. An increase in the frequency of...

  10. 40 CFR 799.9539 - TSCA mammalian erythrocyte micronucleus test.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... as sampled in bone marrow and/or peripheral blood cells of animals, usually rodents. (2) The purpose... immature erythrocytes. (N) Number of cells analyzed per animal. (O) Criteria for considering studies as... micronuclei is facilitated in these cells because they lack a main nucleus. An increase in the frequency of...

  11. 40 CFR 799.9539 - TSCA mammalian erythrocyte micronucleus test.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... as sampled in bone marrow and/or peripheral blood cells of animals, usually rodents. (2) The purpose... immature erythrocytes. (N) Number of cells analyzed per animal. (O) Criteria for considering studies as... micronuclei is facilitated in these cells because they lack a main nucleus. An increase in the frequency of...

  12. 40 CFR 799.9539 - TSCA mammalian erythrocyte micronucleus test.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... sampled in bone marrow and/or peripheral blood cells of animals, usually rodents. (2) The purpose of the... time, unless acceptable inter-animal variability and frequencies of cells with micronuclei are... for scoring micronucleated immature erythrocytes. (N) Number of cells analyzed per animal. (O...

  13. Erythrocyte catalase and carbonic anhydrase activities in lung cancer.

    PubMed

    Cobanoglu, Ufuk; Demir, Halit; Duran, Memet; Şehitogullari, Abidin; Mergan, Duygu; Demir, Canan

    2010-01-01

    To study the relationship between the pathogenesis of lung cancer and antioxidant status and acidic media by measuring the activities of erythrocyte catalase (CAT) and carbonic anhydrase (CA). A total of 26 patients with lung cancer and 15 healthy individuals were included in the study. The CAT and CA activities of erythrocytes were defined. The catalase (CAT) activity of erythrocytes was measured using Aebi's method. Carbonic anhydrase (CA) activity was analyzed by CO2 hydration. It was found that erythrocyte CA and CAT activities were significantly lower in patients with lung cancer compared to controls (p<0.05). Of the 26 patients with lung cancer, seven (26.9%) had metastasis, and the CA and CAT levels in patients with metastasis were significantly decreased (p=0.0001). Development of oxidative stress due to lung cancer may be related to the balance between prooxidant and antioxidant reactions. Catalase may have a preventive effect for malignant lung cancers and the gene of the antioxidant enzymes may be one of the anti-oncogenes, and inactivation of one of these genes in the process of carcinogenesis may lead to tumor development. This may be an explanation for the very low levels of antioxidant CAT in patients with lung cancer compared to healthy individuals. Carbonic anhydrase (CA) in tumor cells may be an indicator of the acid-base balance in lung cancer. Decreased levels of CA in patients with lung cancer may provide a convenient media for tumor development, growth and metastasis by creating an acidic media.

  14. Ceramide in the regulation of eryptosis, the suicidal erythrocyte death.

    PubMed

    Lang, Elisabeth; Bissinger, Rosi; Gulbins, Erich; Lang, Florian

    2015-05-01

    Similar to apoptosis of nucleated cells, erythrocytes may undergo eryptosis, a suicidal death characterized by cell shrinkage and phospholipid scrambling of the cell membrane leading to phosphatidylserine exposure at the cell surface. As eryptotic erythrocytes are rapidly cleared from circulating blood, excessive eryptosis may lead to anemia. Moreover, eryptotic erythrocytes may adhere to the vascular wall and thus impede microcirculation. Stimulators of eryptosis include osmotic shock, oxidative stress and energy depletion. Mechanisms involved in the stimulation eryptosis include ceramide formation which may result from phospholipase A2 dependent formation of platelet activating factor (PAF) with PAF dependent stimulation of sphingomyelinases. Enhanced erythrocytic ceramide formation is observed in fever, sepsis, HUS, uremia, hepatic failure, and Wilson's disease. Enhanced eryptosis is further observed in iron deficiency, phosphate depletion, dehydration, malignancy, malaria, sickle-cell anemia, beta-thalassemia and glucose-6-phosphate dehydrogenase-deficiency. Moreover, eryptosis is triggered by osmotic shock and a wide variety of xenobiotics, which are again partially effective by enhancing ceramide abundance. Ceramide formation is inhibited by high concentrations of urea. As shown in Wilson's disease, pharmacological interference with ceramide formation may be a therapeutic option in the treatment of eryptosis inducing clinical disorders.

  15. Oscillatory tank-treading motion of erythrocytes in shear flows

    PubMed Central

    Dodson, W. R.; Dimitrakopoulos, P.

    2013-01-01

    In this paper, we investigate the oscillatory dynamics of the tank-treading motion of healthy human erythrocytes in shear flows with capillary number Ca = O(1) and small to moderate viscosity ratios 0.01 ≤ λ ≤ 1.5. These conditions correspond to a wide range of surrounding medium viscosities (4 to 600 mPa s) and shear flow rates (2 to 560 s−1), and match those used in ektacytometry systems. For a given viscosity ratio, as the flow rate increases, the steady-state erythrocyte length L (in the shear plane) increases logarithmically while its depth W (normal to the shear plane) decreases logarithmically. In addition, the flow rate increase dampens the oscillatory erythrocyte inclination but not its lengths oscillations (which show relative variations of about 5–8%). For a given flow rate, as the viscosity ratio increases, the erythrocyte length L contracts while its depth W increases (i.e. the cell becomes less deformed) with small decrease in the lengths variations. The average orientation angle of the erythrocyte shows a significant decrease with the viscosity ratio as does the angle oscillation while the oscillation period increases. These trends continue in higher viscosity ratios resulting eventually in the transition from a (weakly oscillatory) tank-treading motion to a tumbling motion. Our computations show that the erythrocyte width S, which exists in the shear plane, is practically invariant in time, capillary number and viscosity ratio, and corresponds to a real cell thickness of about 2.5 μm. Comparison of our computational results with the predictions of (low degree-of-freedom) theoretical models and experimental findings, suggests that the energy dissipation due to the shape-memory effects is more significant than the energy dissipation due to the membrane viscosity. Our work shows that the oscillatory tank-treading motion can account for more than 50% of the variations found in ektacytometry systems; thus, researchers who wish to study inherent

  16. Oscillatory tank-treading motion of erythrocytes in shear flows

    NASA Astrophysics Data System (ADS)

    Dodson, W. R., III; Dimitrakopoulos, P.

    2011-07-01

    In this paper, we investigate the oscillatory dynamics of the tank-treading motion of healthy human erythrocytes in shear flows with capillary number Ca=O(1) and small to moderate viscosity ratios 0.01⩽⩽1.5. These conditions correspond to a wide range of surrounding medium viscosities (4-600 m Pa s) and shear flow rates (2-560s-1), and match those used in ektacytometry systems. For a given viscosity ratio, as the flow rate increases, the steady-state erythrocyte length L (in the shear plane) increases logarithmically while its depth W (normal to the shear plane) decreases logarithmically. In addition, the flow rate increase dampens the oscillatory erythrocyte inclination but not its length oscillations (which show relative variations of about 5-8%). For a given flow rate, as the viscosity ratio increases, the erythrocyte length L contracts while its depth W increases (i.e., the cell becomes less deformed) with a small decrease in the length variations. The average orientation angle of the erythrocyte shows a significant decrease with the viscosity ratio as does the angle oscillation while the oscillation period increases. These trends continue in higher viscosity ratios resulting eventually in the transition from a (weakly oscillatory) tank-treading motion to a tumbling motion. Our computations show that the erythrocyte width S, which exists in the shear plane, is practically invariant in time, capillary number, and viscosity ratio, and corresponds to a real cell thickness of about 2.5μm. Comparison of our computational results with the predictions of (low degree-of-freedom) theoretical models and experimental findings, suggests that the energy dissipation due to the shape-memory effects is more significant than the energy dissipation due to the membrane viscosity. Our work shows that the oscillatory tank-treading motion can account for more than 50% of the variations found in ektacytometry systems; thus, researchers who wish to study inherent differences

  17. Erythrocyte binding preference of human pandemic influenza virus a and its effect on antibody response detection.

    PubMed

    Makkoch, Jarika; Prachayangprecha, Slinporn; Payungporn, Sunchai; Chieochansin, Thaweesak; Songserm, Thaweesak; Amonsin, Alongkorn; Poovorawan, Yong

    2012-07-01

    Validation of hemagglutination inhibition (HI) assays is important for evaluating antibody responses to influenza virus, and selection of erythrocytes for use in these assays is important. This study aimed to determine the correlation between receptor binding specificity and effectiveness of the HI assay for detecting antibody response to pandemic influenza H1N1 (pH1N1) virus. Hemagglutination (HA) tests were performed using erythrocytes from 6 species. Subsequently, 8 hemagglutinating units of pH1N1 from each species were titrated by real-time reverse transcription-PCR. To investigate the effect of erythrocyte binding preference on HI antibody titers, comparisons of HI with microneutralization (MN) assays were performed. Goose erythrocytes showed most specific binding with pH1N1, while HA titers using human erythrocytes were comparable to those using turkey erythrocytes. The erythrocyte binding efficiency was shown to have an impact on antibody detection. Comparing MN titers, HI titers using turkey erythrocytes yielded the most accurate results, while those using goose erythrocytes produced the highest geometric mean titer. Human blood group O erythrocytes lacking a specific antibody yielded results most comparable to those obtained using turkey erythrocytes. Further, pre-existing antibody to pH1N1 and different erythrocyte species can distort HI assay results. HI assay, using turkey and human erythrocytes, yielded the most comparable and applicable results for pH1N1 than those by MN assay, and using goose erythrocytes may lead to overestimated titers. Selection of appropriate erythrocyte species for HI assay allows construction of a more reliable database, which is essential for further investigations and control of virus epidemics.

  18. Maryland Kids Count Factbook, 2001.

    ERIC Educational Resources Information Center

    Advocates for Children and Youth, Baltimore, MD.

    This 7th annual Kids Count Factbook provides information on trends in the well-being of children in Maryland and its 24 jurisdictions. The statistical portrait is based on 18 indicators of well-being: (1) low birth-weight infants; (2) infant mortality; (3) early prenatal care; (4) binge drinking; (5) child deaths; (6) child injury rate; (7) grade…

  19. Counting a Culture of Mealworms

    ERIC Educational Resources Information Center

    Ashbrook, Peggy

    2007-01-01

    Math is not the only topic that will be discussed when young children are asked to care for and count "mealworms," a type of insect larvae (just as caterpillars are the babies of butterflies, these larvae are babies of beetles). The following activity can take place over two months as the beetles undergo metamorphosis from larvae to adults. As the…

  20. Kids Count Data Sheet, 2000.

    ERIC Educational Resources Information Center

    Annie E. Casey Foundation, Baltimore, MD.

    Data from the 50 United States are listed for 1997 from Kids Count in an effort to track state-by-state the status of children in the United States and to secure better futures for all children. Data include percent low birth weight babies; infant mortality rate; child death rate; rate of teen deaths by accident, homicide, and suicide; teen birth…

  1. Oklahoma Kids Count Factbook, 2002.

    ERIC Educational Resources Information Center

    Ingraham, Sandy

    This Kids Count Factbook details county and statewide trends in the well-being of children in Oklahoma. The statistical portrait is based on seven indicators or benchmarks of child well-being: (1) low birthweight infants; (2) infant mortality; (3) births to young teens; (4) child abuse and neglect; (5) child and teen death; (6) high school…

  2. Kids Count in Colorado! 2003.

    ERIC Educational Resources Information Center

    Boeke, Kaye

    This Kids Count report examines state, county, and regional trends in the well-being of Colorado's children. The first part of the report is presented in four chapters. Chapter 1 includes findings regarding the increasing diversity of the child population, linguistic isolation, the impact of parental unemployment, child poverty, and the affordable…

  3. Oklahoma Kids Count Factbook, 2001.

    ERIC Educational Resources Information Center

    Ingraham, Sandy

    This Kids Count Factbook details county and statewide trends in the well-being of Oklahoma's children. The statistical portrait is based on seven indicators or benchmarks of child well-being: (1) low birth weight infants; (2) infant mortality; (3) births to young teens; (4) child abuse and neglect; (5) child and teen death; (6) high school…

  4. Wiskids Count Data Book, 2003.

    ERIC Educational Resources Information Center

    Cranley, M. Martha; Bianchi, J. P.; Eleson, Charity; Hall, Linda; Jacobson, Bob; Jackson, Kristin; Peacock, Jon

    This WisKids Count data book provides a statistical portrait of the well-being of Wisconsin's children. In addition to demographic data indicating changing communities, the indicators and data are organized into five overarching goals: (1) Healthy Families and Children Thrive, including births to single women, infant deaths, and health care…

  5. Are we counting mitoses correctly?

    PubMed

    Yigit, Nuri; Gunal, Armagan; Kucukodaci, Zafer; Karslioglu, Yildirim; Onguru, Onder; Ozcan, Ayhan

    2013-12-01

    The number of mitotic figures in a predefined area is essential in pathologic evaluation for most tumors. This information sometimes provides clues in differentiating neoplastic lesions from nonneoplastic ones and sometimes in defining and grading of the tumors as well as prognosticating expected lifetime of the patient. As a generally accepted concept, scanning a certain number of consecutive nonoverlapping areas that are rich in viable tumor cells is required. Invasion fronts or the periphery of the tumors is preferred for counting mitosis. The target area to be counted for mitotic activity for various tumors is standardized as the number of mitosis in an established number of high-power fields. However, suggested mitotic counts, which constitute the basis of these studies, were obtained via the old microscopes, which usually had narrower visual fields than the state-of-the-art microscopes. Because the visual fields of the present microscopes provide larger areas compared with the older ones, corrections in mitosis counting are needed to make them compatible with the criteria, which had been put forward in the original reference studies.

  6. Shakespeare Live! and Character Counts.

    ERIC Educational Resources Information Center

    Brookshire, Cathy A.

    This paper discusses a live production of Shakespeare's "Macbeth" (in full costume but with no sets) for all public middle school and high school students in Harrisonburg and Rockingham, Virginia. The paper states that the "Character Counts" issues that are covered in the play are: decision making, responsibility and…

  7. Maryland KIDS COUNT Factbook, 1999.

    ERIC Educational Resources Information Center

    Advocates for Children and Youth, Baltimore, MD.

    This Kids Count factbook is the fifth to examine statewide and county trends in the well-being of Maryland's children. The statistical portrait is based on indicators in the domains of economic well-being, good health, safety, and preparing for adulthood. The 16 indicators are: (1) child poverty; (2) child support; (3) births to teens; (4) low…

  8. Wyoming Kids Count Factbook, 1997.

    ERIC Educational Resources Information Center

    Wyoming Kids Count, Cheyenne.

    This Kids Count factbook details statewide trends in the well-being of Wyoming's children. The 1997 report has been expanded to include detailed information on the status of children by categories of welfare, health, and education. The first part of the factbook documents trends by county for 15 indicators: (1) poverty and population; (2)…

  9. Counting a Culture of Mealworms

    ERIC Educational Resources Information Center

    Ashbrook, Peggy

    2007-01-01

    Math is not the only topic that will be discussed when young children are asked to care for and count "mealworms," a type of insect larvae (just as caterpillars are the babies of butterflies, these larvae are babies of beetles). The following activity can take place over two months as the beetles undergo metamorphosis from larvae to adults. As the…

  10. The counting recursive digital filter.

    NASA Technical Reports Server (NTRS)

    Zohar, S.

    1973-01-01

    Analysis of the bit-level operations involved in the convolutions realizing recursive digital filters leads to hardware designs of such filters based on the operation of counting. Various designs realizing both the canonic and 'direct' forms are presented with particular emphasis on low-cost low-speed high-flexibility machines.

  11. Wiskids Count Data Book, 2003.

    ERIC Educational Resources Information Center

    Cranley, M. Martha; Bianchi, J. P.; Eleson, Charity; Hall, Linda; Jacobson, Bob; Jackson, Kristin; Peacock, Jon

    This WisKids Count data book provides a statistical portrait of the well-being of Wisconsin's children. In addition to demographic data indicating changing communities, the indicators and data are organized into five overarching goals: (1) Healthy Families and Children Thrive, including births to single women, infant deaths, and health care…

  12. KIDS COUNT Data Brief, 2009

    ERIC Educational Resources Information Center

    Annie E. Casey Foundation, 2009

    2009-01-01

    This 2009 KIDS COUNT Data Brief features highlights of the enhanced, mobile-friendly Data Center; data on the 10 key indicators of child well-being for all U.S. states, the District of Columbia, and many cities, counties, and school districts; and a summary of this year's essay, which calls for improvements to the nation's ability to design and…

  13. Meal Counting and Claiming Manual.

    ERIC Educational Resources Information Center

    Food and Nutrition Service (USDA), Washington, DC.

    This manual contains information about the selection and implementation of a meal counting and claiming system for the National School Lunch Program (NSLP) and the School Breakfast Program (BSP). Federal reimbursement is provided for each meal that meets program requirements and is served to an eligible student. Part 1 explains the six elements of…

  14. Verbal Counting in Bilingual Contexts

    ERIC Educational Resources Information Center

    Donevska-Todorova, Ana

    2015-01-01

    Informal experiences in mathematics often include playful competitions among young children in counting numbers in as many as possible different languages. Can these enjoyable experiences result with excellence in the formal processes of education? This article discusses connections between mathematical achievements and natural languages within…

  15. KIDS COUNT New Hampshire, 2000.

    ERIC Educational Resources Information Center

    Shemitz, Elllen, Ed.

    This Kids Count report presents statewide trends in the well-being of New Hampshire's children. The statistical report is based on 22 indicators of child well-being in 5 interrelated areas: (1) children and families (including child population, births, children living with single parent, and children experiencing parental divorce); (2) economic…

  16. South Carolina Kids Count, 2000.

    ERIC Educational Resources Information Center

    Holmes, A. Baron

    This Kids Count report examines statewide trends in the well-being of South Carolina's children. The statistical portrait is based on 41 indicators in the areas of demographics, family, economic status, health, readiness and early school performance, scholastic achievement, and adolescent risk behaviors. The indicators are: (1) population; (2)…

  17. South Carolina Kids Count, 2001.

    ERIC Educational Resources Information Center

    Holmes, A. Baron

    This Kids Count report examines statewide trends in the well-being of South Carolina's children. The statistical portrait is based on 42 indicators in the areas of demographics, family, economic status, health, readiness and early school performance, scholastic achievement, and adolescent risk behaviors. The indicators are: (1) population; (2)…

  18. Stimulation of erythrocyte phosphatidylserine exposure by mercury ions

    SciTech Connect

    Eisele, Kerstin; Lang, Philipp A.; Kempe, Daniela S.; Klarl, Barbara A.; Niemoeller, Olivier; Wieder, Thomas; Huber, Stephan M.; Duranton, Christophe; Lang, Florian . E-mail: florian.lang@uni-tuebingen.de

    2006-01-15

    The sequelae of mercury intoxication include induction of apoptosis. In nucleated cells, Hg{sup 2+}-induced apoptosis involves mitochondrial damage. The present study has been performed to elucidate effects of Hg{sup 2+} in erythrocytes which lack mitochondria but are able to undergo apoptosis-like alterations of the cell membrane. Previous studies have documented that activation of a Ca{sup 2+}-sensitive erythrocyte scramblase leads to exposure of phosphatidylserine at the erythrocyte surface, a typical feature of apoptotic cells. The erythrocyte scramblase is activated by osmotic shock, oxidative stress and/or energy depletion which increase cytosolic Ca{sup 2+} activity and/or activate a sphingomyelinase leading to formation of ceramide. Ceramide sensitizes the scramblase to Ca{sup 2+}. The present experiments explored the effect of Hg{sup 2+} ions on erythrocytes. Phosphatidylserine exposure after mercury treatment was estimated from annexin binding as determined in FACS analysis. Exposure to Hg{sup 2+} (1 {mu}M) indeed significantly increased annexin binding from 2.3 {+-} 0.5% (control condition) to 23 {+-} 6% (n = 6). This effect was paralleled by activation of a clotrimazole-sensitive K{sup +}-selective conductance as measured by patch-clamp recordings and by transient cell shrinkage. Further experiments revealed also an increase of ceramide formation by {approx}66% (n = 7) after challenge with mercury (1 {mu}M). In conclusion, mercury ions activate a clotrimazole-sensitive K{sup +}-selective conductance leading to transient cell shrinkage. Moreover, Hg{sup 2+} increases ceramide formation. The observed mechanisms could similarly participate in the triggering of apoptosis in nucleated cells by Hg{sup 2+}.

  19. Erythrocyte Protein 4.1 Binds and Regulates Myosin

    NASA Astrophysics Data System (ADS)

    Pasternack, Gary R.; Racusen, Richard H.

    1989-12-01

    Myosin was recently identified in erythrocytes and was shown to partition both with membrane and cytosolic fractions, suggesting that it may be loosely bound to membranes [Fowler, V. M., Davis, J. Q. & Bennett, V. (1985) J. Cell Biol. 100, 47-55, and Wong, A. J., Kiehart, D. P. & Pollard, T. D. (1985) J. Biol. Chem. 260, 46-49]; however, the molecular basis for this binding was unclear. The present studies employed immobilized monomeric myosin to examine the interaction of myosin with erythrocyte protein 4.1. In human erythrocytes, protein 4.1 binds to integral membrane proteins and mediates spectrin-actin assembly. Protein 4.1 binds to rabbit skeletal muscle myosin with a Kd = 140 nM and a stoichiometry consistent with 1:1 binding. Heavy meromyosin competes for protein 4.1 binding with Ki = 36-54 nM; however, the S1 fragment (the myosin head) competes less efficiently. Affinity chromatography of partial chymotryptic digests of protein 4.1 on immobilized myosin identified a 10-kDa domain of protein 4.1 as the myosin-binding site. In functional studies, protein 4.1 partially inhibited the actin-activated Mg2+-ATPase activity of rabbit skeletal muscle myosin with Ki = 51 nM. Liver cytosolic and erythrocyte myosins preactivated with myosin light-chain kinase were similarly inhibited by protein 4.1. These studies show that protein 4.1 binds, modulates, and thus may regulate myosin. This interaction might serve to generate the contractile forces involved in Mg2+-ATP-dependent shape changes in erythrocytes and may additionally serve as a model for myosin organization and regulation in non-muscle cells.

  20. Red wine activates plasma membrane redox system in human erythrocytes.

    PubMed

    Tedesco, Idolo; Moccia, Stefania; Volpe, Silvestro; Alfieri, Giovanna; Strollo, Daniela; Bilotto, Stefania; Spagnuolo, Carmela; Di Renzo, Massimo; Aquino, Rita P; Russo, Gian Luigi

    2016-01-01

    In the present study, we report that polyphenols present in red wine obtained by a controlled microvinification process are able to protect human erythrocytes from oxidative stress and to activate Plasma Membrane Redox System (PMRS). Human plasma obtained from healthy subjects was incubated in the presence of whole red wine at a concentration corresponding to 9.13-73 μg/ml gallic acid equivalents to verify the capacity to protect against hypochlorous acid (HOCl)-induced plasma oxidation and to minimize chloramine formation. Red wine reduced hemolysis and chloramine formation induced by HOCl of 40 and 35%, respectively. PMRS present on human erythrocytes transfers electrons from intracellular molecules to extracellular electron acceptors. We demonstrated that whole red wine activated PMRS activity in human erythrocytes isolated from donors in a dose-dependent manner with a maximum at about 70-100 μg/ml gallic acid equivalents. We also showed that red wine increased glutathione (GSH) levels and erythrocytic antioxidant capacity, measured by 2,2-diphenyl-1-picrylhydrazyl (DPPH) quenching assay. Furthermore, we reported that GSH played a crucial role in regulating PMRS activity in erythrocytes. In fact, the effect of iodoacetamide, an alkylating agent that induces depletion of intracellular GSH, was completely counteracted by red wine. Bioactive compounds present in red wine, such as gallic acid, resveratrol, catechin, and quercetin were unable to activate PMRS when tested at the concentrations normally present in aged red wines. On the contrary, the increase of PMRS activity was associated with the anthocyanin fraction, suggesting the capacity of this class of compounds to positively modulate PMRS enzymatic activity.

  1. [Ratio of erythrocyte and plasma in massive blood transfusion].

    PubMed

    Wen, Xian-Hui; Liu, Feng-Xia; Zhang, Jun-Hua; Gui, Rong

    2014-06-01

    This study was purposed to explore the suitable ratio between fresh frozen plasma and erythrocyte by retrospective analysis of coagulation in patients with massive blood transfusion. The clinical data of 151 cases with massive blood transfusion from January 2011 to January 2013 were analyzed retrospectively. According to coagulation, patients were divided into coagulation normal group (138 cases) and coagulation dysfunction group (13 cases). Based on the ratio of 1:1 of fresh frozen plasma and erythrocyte, the patients were divided into high plasma group(2:1), medium plasma group (1:1) and low plasma (<1:1) subgroups. Coagulation was detected before and after 24 h of massive blood transfusion. The results showed that prothrombin time (PT), activated partial thromboplastin time (APTT) and thrombin time (TT) were prolonged, fibrinogen (FIB) level decreased significantly (all P < 0.05) in the low plasma subgroup of coagulation normal group after massive blood transfusion 24 h; the high plasma and the medium plasma group of coagulation normal group had no significant changes in coagulation (P > 0.05); prothrombin time, activated partial thromboplastin time, thrombin time and fibrinogen level in the medium plasma and low plasma subgroup of coagulation dysfunction group after massive transfusion was still in abnormal levels (P > 0.05), coagulation function in high plasma subgroup was improved significantly (P < 0.05). It is concluded that the ratio of plasma to erythrocyte should be adjusted according to the patient's coagulation function during massive blood transfusion, the ratio between fresh frozen plasma and erythrocyte is recommended to be 2:1 in patients of coagulation dysfunction in order to improve the patient's coagulation function and to reduce the incidence of adverse event, the ratio of fresh frozen plasma to erythrocyte is recommended to be 1:1 in patients with normal coagulation so as to reduce the dilutional coagulopathy and hypervolemia of blood.

  2. Erythrocyte membrane composition in patients with primary hypercholesterolemia.

    PubMed

    Vayá, A; Martínez Triguero, M; Réganon, E; Vila, V; Martínez Sales, V; Solá, E; Hernández Mijares, A; Ricart, A

    2008-01-01

    There are conflicting results regarding the erythrocyte membrane cholesterol and phospholipid content in patients with primary hypercholesterolemia (PHC), due to methodological problems in obtaining haemoglobin-free ghosts. At the same time, the different units used and the fact that the cholesterol and phospholipids are not expressed in relation with integral protein membrane content, produces contradictory results. We have analysed in 33 patients with PHC (12 male, 31 female) aged 43+/-12 years and in 33 healthy normolipaemic volunteers (9 male, 24 female) aged 43+/-13 years plasma lipids, along with, erythrocyte membrane cholesterol, phospholipids and integral proteins. PHC patients showed increased erythrocyte membrane cholesterol: 0.36+/-0.15 mg/mg when compared with controls: 0.29+/-0.75 mg/mg; p=0.018. Phospholipid membrane content, although higher in the cases, did not reach statistical significance (PHC patients: 0.38+/-0.15 mg/mg vs. 0.33+/-0.72 mg/mg; p=0.098). The cholesterol/phospholipids ratio (Chol/Ph) was 0.99+/-0.22 in PHC patients versus 0.92+/-0.28 in controls; p=0.127. Our results suggest that there is a slight increase in erythrocyte membrane cholesterol in patients with PHC. Given the increasing importance of erythrocyte membrane cholesterol in the stability of the atheroma plaque due its possible contribution to the clinical signs of ischaemic heart disease, it seems relevant to determine this parameter in risk populations. Therefore, a simple and reproducible method needs to be standardised which would enable comparisons between laboratories and facilitate further studies aimed to it as a marker of acute coronary syndromes.

  3. Teaching Emotionally Disturbed Students to Count Feelings.

    ERIC Educational Resources Information Center

    Bartels, Cynthia S.; Calkin, Abigail B.

    The paper describes a program to teach high school students with emotional and behavior problems to count their feelings, thereby improving their self concept. To aid in instruction, a hierarchy was developed which involved four phases: counting tasks completed and tasks not completed, counting independent actions in class, counting perceptions of…

  4. 7 CFR 993.105 - Size count.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 7 Agriculture 8 2010-01-01 2010-01-01 false Size count. 993.105 Section 993.105 Agriculture Regulations of the Department of Agriculture (Continued) AGRICULTURAL MARKETING SERVICE (Marketing Agreements... Administrative Rules and Regulations Definitions § 993.105 Size count. Size count means the count or number of...

  5. 7 CFR 993.502 - Size count.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 7 Agriculture 8 2010-01-01 2010-01-01 false Size count. 993.502 Section 993.502 Agriculture Regulations of the Department of Agriculture (Continued) AGRICULTURAL MARKETING SERVICE (Marketing Agreements... Pack Specification as to Size Definitions § 993.502 Size count. Size count means the count or number of...

  6. Statistical Aspects of Point Count Sampling

    Treesearch

    Richard J. Barker; John R. Sauer

    1995-01-01

    The dominant feature of point counts is that they do not census birds, but instead provide incomplete counts of individuals present within a survey plot. Considering a simple model for point count sampling, we demonstrate that use of these incomplete counts can bias estimators and testing procedures, leading to inappropriate conclusions. A large portion of the...

  7. Evidence for differences in erythrocyte surface receptors for the malarial parasites, Plasmodium falciparum and Plasmodium knowlesi

    PubMed Central

    1977-01-01

    Human erythrocytes lacking various blood group determinants were susceptible to invasion by Plasmodium falciparum including Duffy- negative erythrocytes that are refractory to invasion by Plasmodium knowlesi. Erythrocytes treated with trypsin or neuraminidase had reduced susceptibility of P. falciparum and normal susceptibility to P. knowlesi. Chymotrypsin treatment (0.1 mg/ml) blocked invasion only by P. knowlesi. The differential effect of enzymatic cleavage of determinats from the erythrocyte surface on invasion by these parasites suggests that P. falciparum and P. knowlesi interact with different determinants on the erythrocyte surface. PMID:327014

  8. Comparison of erythrocyte osmotic fragility among ectotherms and endotherms at three temperatures.

    PubMed

    Aldrich, K; Saunders, D K.

    2001-06-01

    1. Comparison of erythrocyte osmotic fragility (EOF) between various ectotherms and endotherms was investigated at 5, 25, and 38 degrees C. 2. We hypothesized that ectotherms might possess erythrocytes whose osmotic fragility would be less affected by temperature than those of endotherms. 3. Ectotherm erythrocytes were much more osmotically resistant than those of endotherms. 4. The EOF of ectotherms and endotherms showed similar responses to temperature. 5. It does not appear that the osmotic fragility of erythrocytes from ectotherms in this study are adapted to be less affected by temperature than those of endotherms. The highly osmotic resistant erythrocytes of ectotherms may alleviate the need for further adaptation for osmotic resistance.

  9. Predictive Model Assessment for Count Data

    DTIC Science & Technology

    2007-09-05

    critique count regression models for patent data, and assess the predictive performance of Bayesian age-period-cohort models for larynx cancer counts...the predictive performance of Bayesian age-period-cohort models for larynx cancer counts in Germany. We consider a recent suggestion by Baker and...Figure 5. Boxplots for various scores for patent data count regressions. 11 Table 1 Four predictive models for larynx cancer counts in Germany, 1998–2002

  10. Acute dark chocolate ingestion is beneficial for hemodynamics via enhancement of erythrocyte deformability in healthy humans.

    PubMed

    Radosinska, Jana; Horvathova, Martina; Frimmel, Karel; Muchova, Jana; Vidosovicova, Maria; Vazan, Rastislav; Bernatova, Iveta

    2017-03-01

    Erythrocyte deformability is an important property of erythrocytes that considerably affects blood flow and hemodynamics. The high content of polyphenols present in dark chocolate has been reported to play a protective role in functionality of erythrocytes. We hypothesized that chocolate might influence erythrocytes not only after repeated chronic intake, but also immediately after its ingestion. Thus, we determined the acute effect of dark chocolate and milk (with lower content of biologically active substances) chocolate intake on erythrocyte deformability. We also focused on selected factors that may affect erythrocyte deformability, specifically nitric oxide production in erythrocytes and total antioxidant capacity of plasma. We determined posttreatment changes in the mentioned parameters 2hours after consumption of chocolate compared with their levels before consumption of chocolate. In contrast to milk chocolate intake, the dark chocolate led to a significantly higher increase in erythrocyte deformability. Nitric oxide production in erythrocytes was not changed after dark chocolate intake, but significantly decreased after milk chocolate. The plasma total antioxidant capacity remained unaffected after ingestion of both chocolates. We conclude that our hypothesis was confirmed. Single ingestion of dark chocolate improved erythrocyte deformability despite unchanged nitric oxide production and antioxidant capacity of plasma. Increased deformability of erythrocytes may considerably improve rheological properties of blood and thus hemodynamics in humans, resulting in better tissue oxygenation. Copyright © 2017 Elsevier Inc. All rights reserved.

  11. Attempts to validate a possible predictive animal model for human erythrocyte G-6-PD deficiency

    SciTech Connect

    Horton, H.M.; Calabrese, E.J.

    1986-01-01

    The use of Dorset sheep erythrocytes as a model for human G-6-PD deficient erythrocytes was investigated. Seven pharmaceuticals were examined for oxidant stressor effects using a liver microsomal enzyme system to generate metabolites of the drugs. The pharmaceuticals examined were salicyclic acid, dapsone, naphthalene, B-naphtol, p-aminobenzoic acid, sulfanilamide and sulfapyridine. The test compounds were incubated with Dorset sheep erythrocytes and oxidant stressor effects were measured through reduced glutathione (GSH) levels and methemaglobin formation. The response of the Dorset sheep erythrocytes to the seven agents was compared to previous studies revealing the response of human G-6-PD deficient erythrocytes to these agents. The results indicated that metabolites of the pharmaceuticals, B-naphthol, dapsone, and sulfanilamide, are oxidant stressor agents towards sheep G-6-PD deficient erythrocytes. These results agreed with studies on the response of human G-6-PD deficient erythrocytes. The metabolized naphthalene and sulfapyridine did not cause oxidant stress in the sheep erythrocytes, despite the fact that these two agents caused oxidizing effects in human G-6-PD deficient erythrocytes in previous studies. None of the non-metabolized parent compounds caused oxidant stress in the sheep erythrocytes, which agreed with the responses of human G-6-PD deficient erythrocytes.

  12. Response of the iron-deficient erythrocyte in the rat to hyperoxia

    NASA Technical Reports Server (NTRS)

    Larkin, E. C.; Kimzey, S. L.; Siler, K.

    1978-01-01

    Normal and iron-deficient rats were exposed to 90% O2 at 760 Torr for 24 or 48 h. Erythrocyte response to hyperoxia was monitored by potassium (rubidium) influx studies, by storage stress, and by ultrastructural studies. Normal rat erythrocytes exhibited morphological changes and decrease of ouabain-sensitive potassium influx compared to unexposed controls. Both components of erythrocyte potassium influx were affected by iron deficiency. Erythrocytes from unexposed iron-deficient rats showed a 50% increase in ouabain-sensitive potassium influx compared to controls. Iron-deficient rats exposed to hyperoxia for 24 or 48 h, had erythrocytes with morphological changes. Erythrocytes of iron-deficient rats exposed for 24 h showned no influx change; those exposed for 48 h showed a decrease of ouabain-sensitive influx compared to erythrocytes of controls.

  13. Response of the iron-deficient erythrocyte in the rat to hyperoxia

    NASA Technical Reports Server (NTRS)

    Larkin, E. C.; Kimzey, S. L.; Siler, K.

    1978-01-01

    Normal and iron-deficient rats were exposed to 90% O2 at 760 Torr for 24 or 48 h. Erythrocyte response to hyperoxia was monitored by potassium (rubidium) influx studies, by storage stress, and by ultrastructural studies. Normal rat erythrocytes exhibited morphological changes and decrease of ouabain-sensitive potassium influx compared to unexposed controls. Both components of erythrocyte potassium influx were affected by iron deficiency. Erythrocytes from unexposed iron-deficient rats showed a 50% increase in ouabain-sensitive potassium influx compared to controls. Iron-deficient rats exposed to hyperoxia for 24 or 48 h, had erythrocytes with morphological changes. Erythrocytes of iron-deficient rats exposed for 24 h showned no influx change; those exposed for 48 h showed a decrease of ouabain-sensitive influx compared to erythrocytes of controls.

  14. Genetic Evidence for Erythrocyte Receptor Glycophorin B Expression Levels Defining a Dominant Plasmodium falciparum Invasion Pathway into Human Erythrocytes

    PubMed Central

    Dankwa, Selasi; Chaand, Mudit; Kanjee, Usheer; Jiang, Rays H. Y.; Nobre, Luis V.; Goldberg, Jonathan M.; Bei, Amy K.; Moechtar, Mischka A.; Grüring, Christof; Ahouidi, Ambroise D.; Ndiaye, Daouda; Dieye, Tandakha N.; Mboup, Souleymane; Weekes, Michael P.

    2017-01-01

    ABSTRACT Plasmodium falciparum, the parasite that causes the deadliest form of malaria, has evolved multiple proteins known as invasion ligands that bind to specific erythrocyte receptors to facilitate invasion of human erythrocytes. The EBA-175/glycophorin A (GPA) and Rh5/basigin ligand-receptor interactions, referred to as invasion pathways, have been the subject of intense study. In this study, we focused on the less-characterized sialic acid-containing receptors glycophorin B (GPB) and glycophorin C (GPC). Through bioinformatic analysis, we identified extensive variation in glycophorin B (GYPB) transcript levels in individuals from Benin, suggesting selection from malaria pressure. To elucidate the importance of the GPB and GPC receptors relative to the well-described EBA-175/GPA invasion pathway, we used an ex vivo erythrocyte culture system to decrease expression of GPA, GPB, or GPC via lentiviral short hairpin RNA transduction of erythroid progenitor cells, with global surface proteomic profiling. We assessed the efficiency of parasite invasion into knockdown cells using a panel of wild-type P. falciparum laboratory strains and invasion ligand knockout lines, as well as P. falciparum Senegalese clinical isolates and a short-term-culture-adapted strain. For this, we optimized an invasion assay suitable for use with small numbers of erythrocytes. We found that all laboratory strains and the majority of field strains tested were dependent on GPB expression level for invasion. The collective data suggest that the GPA and GPB receptors are of greater importance than the GPC receptor, supporting a hierarchy of erythrocyte receptor usage in P. falciparum. PMID:28760933

  15. Fatty acids of erythrocyte membrane in acute pancreatitis patients

    PubMed Central

    Kuliaviene, Irma; Gulbinas, Antanas; Cremers, Johannes; Pundzius, Juozas; Kupcinskas, Limas; Dambrauskas, Zilvinas; Jansen, Eugene

    2013-01-01

    AIM: To evaluate changes in the fatty acid composition of erythrocyte membrane phospholipids during severe and mild acute pancreatitis (AP) of alcoholic and nonalcoholic etiology. METHODS: All consecutive patients with a diagnosis of AP and onset of the disease within the last 72 h admitted to the Hospital of Lithuanian University of Health Sciences between June and December 2007 were included. According to the Acute Physiology and Chronic Health Evaluation (APACHE II) scale, the patients were subdivided into the mild (APACHE II score < 7, n = 22) and severe (APACHE II score ≥ 7, n = 17) AP groups. Healthy individuals (n = 26) were enrolled as controls. Blood samples were collected from patients on admission to the hospital. Fatty acids (FAs) were extracted from erythrocyte phospholipids and expressed as percentages of the total FAs present in the chromatogram. The concentrations of superoxide dismutase and glutathione peroxidase were measured in erythrocytes. RESULTS: We found an increase in the percentages of saturated and monounsaturated FAs, a decrease in the percentages of total polyunsaturated FAs (PUFAs) and n-3 PUFAs in erythrocyte membrane phospholipids of AP patients compared with healthy controls. Palmitic (C16:0), palmitoleic (C16:1n7cis), arachidonic (C20:4n6), docosahexaenoic (DHA, C22:6n3), and docosapentaenoic (DPA, C22:5n3) acids were the major contributing factors. A decrease in the peroxidation and unsaturation indexes in AP patients as well as the severe and mild AP groups as compared with controls was observed. The concentrations of antioxidant enzymes in the mild AP group were lower than in the control group. In severe AP of nonalcoholic etiology, the percentages of arachidic (C20:0) and arachidonic (C20:4n6) acids were decreased as compared with the control group. The patients with mild AP of nonalcoholic etiology had the increased percentages of total saturated FAs and gama linoleic acid (C18:3n6) and the decreased percentages of elaidic

  16. Fatty acids of erythrocyte membrane in acute pancreatitis patients.

    PubMed

    Kuliaviene, Irma; Gulbinas, Antanas; Cremers, Johannes; Pundzius, Juozas; Kupcinskas, Limas; Dambrauskas, Zilvinas; Jansen, Eugene

    2013-09-14

    To evaluate changes in the fatty acid composition of erythrocyte membrane phospholipids during severe and mild acute pancreatitis (AP) of alcoholic and nonalcoholic etiology. All consecutive patients with a diagnosis of AP and onset of the disease within the last 72 h admitted to the Hospital of Lithuanian University of Health Sciences between June and December 2007 were included. According to the Acute Physiology and Chronic Health Evaluation (APACHE II) scale, the patients were subdivided into the mild (APACHE II score < 7, n = 22) and severe (APACHE II score ≥ 7, n = 17) AP groups. Healthy individuals (n = 26) were enrolled as controls. Blood samples were collected from patients on admission to the hospital. Fatty acids (FAs) were extracted from erythrocyte phospholipids and expressed as percentages of the total FAs present in the chromatogram. The concentrations of superoxide dismutase and glutathione peroxidase were measured in erythrocytes. We found an increase in the percentages of saturated and monounsaturated FAs, a decrease in the percentages of total polyunsaturated FAs (PUFAs) and n-3 PUFAs in erythrocyte membrane phospholipids of AP patients compared with healthy controls. Palmitic (C16:0), palmitoleic (C16:1n7cis), arachidonic (C20:4n6), docosahexaenoic (DHA, C22:6n3), and docosapentaenoic (DPA, C22:5n3) acids were the major contributing factors. A decrease in the peroxidation and unsaturation indexes in AP patients as well as the severe and mild AP groups as compared with controls was observed. The concentrations of antioxidant enzymes in the mild AP group were lower than in the control group. In severe AP of nonalcoholic etiology, the percentages of arachidic (C20:0) and arachidonic (C20:4n6) acids were decreased as compared with the control group. The patients with mild AP of nonalcoholic etiology had the increased percentages of total saturated FAs and gama linoleic acid (C18:3n6) and the decreased percentages of elaidic (C18:1n9t

  17. Tank-treading of swollen erythrocytes in shear flows

    NASA Astrophysics Data System (ADS)

    Dodson, W. R., III; Dimitrakopoulos, P.

    2012-02-01

    In this paper, we investigate computationally the oscillatory tank-treading motion of healthy swollen human erythrocytes (owing to lower than physiological plasma osmolarity) in shear flows with capillary number Ca=O(1) and small to moderate viscosity ratios 0.01≤λ≤2.75. Swollen cells show similar shear flow dynamics with normal cells but with significantly higher inclination and tank-treading speed owing to the higher cell thickness. For a given viscosity ratio, as the flow rate increases, the steady-state erythrocyte length L (in the shear plane) increases logarithmically while its depth W (normal to the shear plane) decreases logarithmically; increase of the viscosity ratio results in lower cell deformation. The erythrocyte width S, which exists in the shear plane, is practically invariant in time, flow rate, and viscosity ratio and corresponds to a real cell thickness of about 2.5μm at physiological osmolarity (300mO) and 3.4μm at an osmolarity of 217 mO. The erythrocyte inclination decreases as the flow rate increases or as the surrounding fluid viscosity decreases, owing to the increased inner rotational flow which tends to align the cell toward the flow direction. The ektacytometry deformation of swollen cells increases logarithmically with the shear stress but with a slower slope than that for normal cells owing mainly to the higher orientation of the more swollen cells. As the cell swelling increases, the tank-treading period decreases owing to the higher thickness of the actual cell which overcomes the opposite action of the reduced shape-memory effects (i.e., the more spherical-like erythrocyte's reference shape of shearing resistance). The local area incompressibility tensions from the lipid bilayer increase with the cell swelling and cause a higher cytoskeleton prestress; this increased prestress results in smaller, but still measurable, local area changes on the spectrin skeleton of the more swollen erythrocytes. Our work provides insight on

  18. Tank-treading of swollen erythrocytes in shear flows.

    PubMed

    Dodson, W R; Dimitrakopoulos, P

    2012-02-01

    In this paper, we investigate computationally the oscillatory tank-treading motion of healthy swollen human erythrocytes (owing to lower than physiological plasma osmolarity) in shear flows with capillary number Ca=O(1) and small to moderate viscosity ratios 0.01≤λ≤2.75. Swollen cells show similar shear flow dynamics with normal cells but with significantly higher inclination and tank-treading speed owing to the higher cell thickness. For a given viscosity ratio, as the flow rate increases, the steady-state erythrocyte length L (in the shear plane) increases logarithmically while its depth W (normal to the shear plane) decreases logarithmically; increase of the viscosity ratio results in lower cell deformation. The erythrocyte width S, which exists in the shear plane, is practically invariant in time, flow rate, and viscosity ratio and corresponds to a real cell thickness of about 2.5 μm at physiological osmolarity (300 mO) and 3.4 μm at an osmolarity of 217 mO. The erythrocyte inclination decreases as the flow rate increases or as the surrounding fluid viscosity decreases, owing to the increased inner rotational flow which tends to align the cell toward the flow direction. The ektacytometry deformation of swollen cells increases logarithmically with the shear stress but with a slower slope than that for normal cells owing mainly to the higher orientation of the more swollen cells. As the cell swelling increases, the tank-treading period decreases owing to the higher thickness of the actual cell which overcomes the opposite action of the reduced shape-memory effects (i.e., the more spherical-like erythrocyte's reference shape of shearing resistance). The local area incompressibility tensions from the lipid bilayer increase with the cell swelling and cause a higher cytoskeleton prestress; this increased prestress results in smaller, but still measurable, local area changes on the spectrin skeleton of the more swollen erythrocytes. Our work provides insight on

  19. Structural alterations of the erythrocyte membrane proteins in diabetic retinopathy.

    PubMed

    Petropoulos, Ioannis K; Margetis, Panagiotis I; Antonelou, Marianna H; Koliopoulos, John X; Gartaganis, Sotirios P; Margaritis, Lukas H; Papassideri, Issidora S

    2007-08-01

    Several rheological disorders of the erythrocytes, such as increased aggregation and decreased deformability, have been observed in diabetes mellitus and have been implicated in the development of diabetic microangiopathy. Structural alterations of the erythrocyte membrane proteins caused by the diabetic process may be at the origin of those observations. In the present study, we searched for erythrocyte membrane protein alterations in diabetic retinopathy. We examined peripheral blood samples from 40 type-2 diabetic patients with diabetic retinopathy of variable severity (19 males and 21 females, mean age 66.8 years, Group A) and we compared them with samples from 19 type-2 diabetic patients without diabetic retinopathy (13 males and six females, mean age 66.5 years, Group B) and 16 healthy volunteers (eight males and eight females, mean age 65.6 years, Group C). Erythrocyte membrane ghosts from all samples were subjected to SDS-PAGE, and the electrophoretic pattern of transmembrane and cytoskeletal proteins was analysed for each sample. The protein quantification of each electrophoretic band was accomplished through scanning densitometry. No significant deviations from normal electrophoresis were observed in Groups B and C, apart from an increase in band 8 in two samples from Group B (11%). In contrast, in 14 samples from Group A (35%) we detected increases in protein band 8 and/or membrane-bound haemoglobin along with a decrease in spectrin. Moreover, increased mobility of band 3, an aberrant high molecular weight (MW) (> 255 kDa) band and a low MW (42 kDa) band were evident in ten samples from Group A (25%). Glycophorins were altered in 46% of Group-A patients versus 38% of Group-B patients. Females and patients with long duration of diabetes presented more electrophoretic abnormalities. Structural alterations of the erythrocyte membrane proteins are shown for the first time in association with diabetic retinopathy. Their detection may serve as a blood marker

  20. Constraint counting for frictional jamming

    NASA Astrophysics Data System (ADS)

    Quint, D. A.; Henkes, S.; Schwarz, J. M.

    2012-02-01

    While the frictionless jamming transition has been intensely studied in recent years, more realistic frictional packings are less well understood. In frictionless sphere packings, the transition is predicted by a simple mean-field constraint counting argument, the isostaticity argument. For frictional packings, a modified constraint counting argument, which includes slipping contacts at the Coulomb threshold, has had limited success in accounting for the transition. We propose that the frictional jamming transition is not mean field and is triggered by the nucleation of unstable regions, which are themselves dynamical objects due to the Coulomb criterion. We create frictional packings using MD simulations and test for the presence and shape of rigid clusters with the pebble game to identify the partition of the packing into stable and unstable regions. To understand the dynamics of these unstable regions we follow perturbations at contacts crucial to the stability of the ``frictional house of cards.''

  1. [Platelet count in the cat].

    PubMed

    Moritz, A; Hoffmann, C

    1997-11-01

    The technique of collecting blood samples is primarily responsible for the appearance of platelet-agglomeration in cats. Blood obtained by the conventional way ("one syringe technology", drips of blood) caused in 52% of the cases an activation of the large and therefore active thrombocytes however. Rejection of the first 2-5 ml blood for the platelet count ("two syringe technology") reduced the rate of platelet-agglomeration significantly. No big differences in platelet-agglomeration were found with regard to the place used for collecting blood (V. cephalica antebrachii/V. jugularis). Platelet-agglutination was observed with Li-Heparin, K-EDTA, Na-Citrat or ACD anticoagulated blood samples. Citrat (Na-Citrat, ACD) seemed to have a stabilizing effect on feline thrombocytes as has been described for human thrombocytes. The platelet count in cats should be performed within 30 minutes.

  2. Bayesian Kernel Mixtures for Counts.

    PubMed

    Canale, Antonio; Dunson, David B

    2011-12-01

    Although Bayesian nonparametric mixture models for continuous data are well developed, there is a limited literature on related approaches for count data. A common strategy is to use a mixture of Poissons, which unfortunately is quite restrictive in not accounting for distributions having variance less than the mean. Other approaches include mixing multinomials, which requires finite support, and using a Dirichlet process prior with a Poisson base measure, which does not allow smooth deviations from the Poisson. As a broad class of alternative models, we propose to use nonparametric mixtures of rounded continuous kernels. An efficient Gibbs sampler is developed for posterior computation, and a simulation study is performed to assess performance. Focusing on the rounded Gaussian case, we generalize the modeling framework to account for multivariate count data, joint modeling with continuous and categorical variables, and other complications. The methods are illustrated through applications to a developmental toxicity study and marketing data. This article has supplementary material online.

  3. Counting Heron Triangles with Constraints

    DTIC Science & Technology

    2013-01-25

    A3 INTEGERS 13 (2013) COUNTING HERON TRIANGLES WITH CONSTRAINTS Pantelimon Stănică Applied Mathematics, Naval Postgraduate School, Monterey...12, Revised: 10/12/12, Accepted: 1/13/13, Published: 1/25/13 Abstract Heron triangles have the property that all three of their sides as well as their...area are positive integers. In this paper, we give some estimates for the number of Heron triangles with two of their sides fixed. We provide a

  4. Characterization of pig lymphocyte receptors for allogeneic and non-allogeneic erythrocytes. I. Apparent common identity of both receptors.

    PubMed Central

    Salmon, H

    1982-01-01

    In the pig thymus, the proportion of allogeneic (or autologous) erythrocyte rosette forming cells (P-RFC) is always lower than that of sheep erythrocyte (non-allogeneic) rosette forming cells (S-RFC) even under saturated RBC/lymphocyte ratios and optimal dextran concentration. This difference accounted for lymphocytes rosetting with sheep erythrocytes and not with pig erythrocytes (P-S+ cells), as opposed to those lymphocytes which are able to bind both types of erythrocytes (P+S+ cells). Since formation of both sheep and pig erythrocyte rosettes is inhibited similarly by anti-T receptor serum, is inhibited reciprocally by sheep and pig erythrocyte membrane fragment and is similarly trypsin sensitive, it was concluded that the same receptor was responsible for both sheep and pig rosette formation. Furthermore it was found that P+S+ cells had a higher avidity for sheep erythrocytes (and lower for pig erythrocytes) than the other subset which did not bind pig erythrocytes. PMID:6180852

  5. Therapeutic efficacies of Coriandrum sativum aqueous extract against metronidazole-induced genotoxicity in Channa punctatus peripheral erythrocytes.

    PubMed

    Talapatra, Soumendra Nath; Dasgupta, Subham; Guha, Gunjan; Auddy, Moumita; Mukhopadhyay, Aniruddha

    2010-12-01

    Metronidazole (MTZ), a nitroimidazole drug, is primarily used as an anti-protozoan or an anti-bacterial agent in humans, although its genotoxic and carcinogenic effects have been widely reported, particularly in aquatic organisms. MTZ may induce DNA damages through single-strand breaks, modification of bases, DNA-DNA and DNA-protein cross-links, ultimately leading to apoptosis or necrosis. Here, we have assessed the genotoxicity of MTZ in the peripheral erythrocytes of Channa punctatus, using micronucleation (MN) and binucleation (BN) as genotoxicity markers. The therapeutic potential of aqueous extract of Coriandrum sativum against MTZ-induced genotoxicity has also been examined. The results show significant (P<0.05) increase in both MN and BN formation due to MTZ treatment. Such aberrations were higher in smaller fish samples for a particular dosage of MTZ, as established by correlation analysis between fish body weight and MN/BN count at P<0.05. However, such degenerative damages were found to be alleviated by a great extent due to treatment with C. sativum leaf extract. Hence, we establish that MTZ can produce considerable degrees of micronucleus and binucleus formation in peripheral erythrocytes of C. punctatus, and such deleterious effect of MTZ treatment can be mitigated by aqueous extract of C. sativum leaves.

  6. Manual and automated reticulocyte counts.

    PubMed

    Simionatto, Mackelly; de Paula, Josiane Padilha; Chaves, Michele Ana Flores; Bortoloso, Márcia; Cicchetti, Domenic; Leonart, Maria Suely Soares; do Nascimento, Aguinaldo José

    2010-12-01

    Manual reticulocyte counts were examined under light microscopy, using the property whereby supravital stain precipitates residual ribosomal RNA versus the automated flow methods, with the suggestion that in the latter there is greater precision and an ability to determine both mature and immature reticulocyte fractions. Three hundred and forty-one venous blood samples of patients were analyzed of whom 224 newborn and the rest adults; 51 males and 66 females, with ages between 0 and 89 years, as part of the laboratory routine for hematological examinations at the Clinical Laboratory of the Hospital Universitário do Oeste do Paraná. This work aimed to compare manual and automated methodologies for reticulocyte countings and evaluate random and systematic errors. The results obtained showed that the difference between the two methods was very small, with an estimated 0·4% systematic error and 3·9% random error. Thus, it has been confirmed that both methods, when well conducted, can reflect precisely the reticulocyte counts for adequate clinical use.

  7. Approximate Counting of Graphical Realizations.

    PubMed

    Erdős, Péter L; Kiss, Sándor Z; Miklós, István; Soukup, Lajos

    2015-01-01

    In 1999 Kannan, Tetali and Vempala proposed a MCMC method to uniformly sample all possible realizations of a given graphical degree sequence and conjectured its rapidly mixing nature. Recently their conjecture was proved affirmative for regular graphs (by Cooper, Dyer and Greenhill, 2007), for regular directed graphs (by Greenhill, 2011) and for half-regular bipartite graphs (by Miklós, Erdős and Soukup, 2013). Several heuristics on counting the number of possible realizations exist (via sampling processes), and while they work well in practice, so far no approximation guarantees exist for such an approach. This paper is the first to develop a method for counting realizations with provable approximation guarantee. In fact, we solve a slightly more general problem; besides the graphical degree sequence a small set of forbidden edges is also given. We show that for the general problem (which contains the Greenhill problem and the Miklós, Erdős and Soukup problem as special cases) the derived MCMC process is rapidly mixing. Further, we show that this new problem is self-reducible therefore it provides a fully polynomial randomized approximation scheme (a.k.a. FPRAS) for counting of all realizations.

  8. Approximate Counting of Graphical Realizations

    PubMed Central

    2015-01-01

    In 1999 Kannan, Tetali and Vempala proposed a MCMC method to uniformly sample all possible realizations of a given graphical degree sequence and conjectured its rapidly mixing nature. Recently their conjecture was proved affirmative for regular graphs (by Cooper, Dyer and Greenhill, 2007), for regular directed graphs (by Greenhill, 2011) and for half-regular bipartite graphs (by Miklós, Erdős and Soukup, 2013). Several heuristics on counting the number of possible realizations exist (via sampling processes), and while they work well in practice, so far no approximation guarantees exist for such an approach. This paper is the first to develop a method for counting realizations with provable approximation guarantee. In fact, we solve a slightly more general problem; besides the graphical degree sequence a small set of forbidden edges is also given. We show that for the general problem (which contains the Greenhill problem and the Miklós, Erdős and Soukup problem as special cases) the derived MCMC process is rapidly mixing. Further, we show that this new problem is self-reducible therefore it provides a fully polynomial randomized approximation scheme (a.k.a. FPRAS) for counting of all realizations. PMID:26161994

  9. The MIC photon counting detector

    NASA Astrophysics Data System (ADS)

    Fordham, J. L. A.; Bone, D. A.; Oldfield, M. K.; Bellis, J. G.; Norton, T. J.

    1992-12-01

    The MIC (Microchannel plate Intensified CCD (Charge Coupled Device)) detector is an advanced performance Micro Channel Plate (MCP) intensified CCD photon counting detector developed for high resolution, high dynamic range, astronomical applications. The heart of the detector is an MCP intensifier developed specifically for photon counting applications. The maximum detector format is 3072 by 2304 pixels. The measured resolution of the detector system is 18 micrometers FWHM at 490 nm. The detector is linear to approximately 1,000,000 events/detector area/sec on a flat field and linear to count rates up to 200 events/object/s on star images. Two versions of the system have been developed. The first for ground based astronomical applications based around a 40 mm diameter intensifier, was proven in trials at a number of large optical telescopes. The second, specifically for the ESA X-Ray Multi Mirror Mission (XMM), where the MIC has been accepted as the blue detector for the incorporated Optical Monitor (OM). For the XMM-OM, the system is based around a 25 mm diameter intensifier. At present, under development, is a 75 mm diameter version of the detector which will have a maximum format of 6144 by 4608 pixels. Details of the MIC detector and its performance are presented.

  10. Differential leukocyte counting and immunophenotyping in cryopreserved ex vivo whole blood.

    PubMed

    Nemes, Elisa; Kagina, Benjamin M N; Smit, Erica; Africa, Hadn; Steyn, Marcia; Hanekom, Willem A; Scriba, Thomas J

    2015-02-01

    Absolute cell counts are typically measured in fresh samples, but this is impractical in large field studies. We compared quantification of leukocyte proportions and absolute counts using reference real-time methods (stain and lyse/no-wash (LNW) or hematology analyser) with a novel assay that allows long-term cryopreservation of fixed leukocytes for later counting (DLC-ICE: differential leukocyte count and immunophenotype in cryopreserved ex vivo whole blood). For the LNW method, whole blood (WB) was stained with fluorescent antibodies, then erythrocytes were lysed, and leukocytes fixed prior to flow cytometry. Alternatively, our novel DLC-ICE method entailed erythrocyte lysis and leukocyte fixation, cryopreservation and later staining of permeabilized cells prior to flow cytometry. Outcomes were proportions and absolute counts of granulocytes, lymphocytes, monocytes, T cells, B cells, and activated T cells within the leukocyte population. We also compared leukocyte subset counts in fresh WB from 51 healthy infants measured by hematology analyser at a rural clinical site or by DLC-ICE method after 2 years of cryopreservation. We observed excellent agreement and strong correlations between absolute counts or cell proportions measured by the LNW and DLC-ICE methods on fresh WB from 10 healthy adults. Compared to LNW, DLC-ICE yielded similar or brighter staining even after cryopreservation. Duration of cryopreservation, assessed monthly for 1 year, had little effect on cell enumeration: median coefficients of variation were below 15% for all outcomes. Under field site conditions, we observed strong correlations between infant leukocyte numbers measured in fresh samples by hematology analyser and those measured by DLC-ICE up to 2 years of cryopreservation. Our novel DLC-ICE method allows accurate flow cytometric quantification of cell subsets from fixed WB even after long-term cryopreservation. This method is ideal for batched, retrospective analysis of samples from

  11. LIN28A expression reduces sickling of cultured human erythrocytes.

    PubMed

    de Vasconcellos, Jaira F; Fasano, Ross M; Lee, Y Terry; Kaushal, Megha; Byrnes, Colleen; Meier, Emily R; Anderson, Molly; Rabel, Antoinette; Braylan, Raul; Stroncek, David F; Miller, Jeffery L

    2014-01-01

    Induction of fetal hemoglobin (HbF) has therapeutic importance for patients with sickle cell disease (SCD) and the beta-thalassemias. It was recently reported that increased expression of LIN28 proteins or decreased expression of its target let-7 miRNAs enhances HbF levels in cultured primary human erythroblasts from adult healthy donors. Here LIN28A effects were studied further using erythrocytes cultured from peripheral blood progenitor cells of pediatric subjects with SCD. Transgenic expression of LIN28A was accomplished by lentiviral transduction in CD34(+) sickle cells cultivated ex vivo in serum-free medium. LIN28A over-expression (LIN28A-OE) increased HbF, reduced beta (sickle)-globin, and strongly suppressed all members of the let-7 family of miRNAs. LIN28A-OE did not affect erythroblast differentiation or prevent enucleation, but it significantly reduced or ameliorated the sickling morphologies of the enucleated erythrocytes.

  12. Recent advancements in erythrocytes, platelets, and albumin as delivery systems

    PubMed Central

    Xu, Peipei; Wang, Ruju; Wang, Xiaohui; Ouyang, Jian

    2016-01-01

    In the past few years, nanomaterial-based drug delivery systems have been applied to enhance the efficacy of therapeutics and to alleviate negative effects through the controlled delivery of targeting and releasing agents. However, few drug carriers can achieve high targeting efficacy, even when targeting modalities and surface markers are introduced. Immunological problems have also limited their wide applications. Biological drug delivery systems, such as erythrocytes, platelets, and albumin, have been extensively investigated because of their unique properties. In this review, erythrocytes, platelets, and albumin are described as efficient drug delivery systems. Their properties, applications, advantages, and limitations in disease treatment are explained. This review confirms that these systems can be used to facilitate a specific, biocompatible, and smart drug delivery. PMID:27274282

  13. [Recent advance on blood group antigen modification of porcine erythrocytes].

    PubMed

    Wang, Jie-Xi; Zhang, Yang-Pei

    2002-06-01

    Advances in the field of xenotransplantation raise the intriguing possibility of using porcine red blood cells (pRBCs) as an alternative source for blood transfusion. Serologically, pRBCs share a number of characteristics with human red blood cells (RBCs), so pRBCs are considered the most likely donor for xenotransfusion. However, xenoantigens on porcine erythrocytes play major roles in antibody-mediated RBC destruction. Although the alphaGal epitope (Galalpha1, 3Galbeta1, 4GalNAc-R) is the major xenoantigen on porcine erythrocytes and is responsible for the binding of the majority of human natural antibodies, other non-alphaGal xenoantigens have been identified. The importance of these non-alphaGal xenoantigens in binding human natural antibodies and subsequently triggering immunological responses cannot be underestimated.

  14. Erythrocyte enzymes catalyze 1-nitropyrene and 3-nitrofluoranthene nitroreduction.

    PubMed

    Belisario, M A; Pecce, R; Garofalo, A; Sannolo, N; Malorni, A

    1996-04-15

    Nitroarenes are environmental contaminants produced during incomplete combustion processes. Nitroreduction, the most important pathway of nitroarene toxification, occurs mainly in the liver and intestine. In the present study, we show that human red cells may also possess the metabolic competence to reduce 1-nitropyrene (NP) and 3-nitrofluoranthene (NF), the nitroarenes chosen as model compounds, to their corresponding amino derivatives, 1-aminopyrene (AP) and 3-aminofluoranthene (AF). The requirement of the cofactor couple NADH/FMN suggests that erythrocyte nitroreductase activity occurs via one electron transfer. The presence of oxygen strongly inhibited the haemolysate-catalyzed nitroarene reduction, whether measured as amine formation or nitroarene disappearance. Intermediate reactive species, that bind covalently to haemoglobin and/or other erythrocyte proteins, are formed during nitroreduction catalyzed by human haemolysate. In fact, the reduced metabolites AP and AF were released after mild acid hydrolysis of red cell proteins exposed to NP and NF, thus suggesting that sulphinamide adducts have been formed.

  15. Why do malaria parasites increase host erythrocyte permeability?

    PubMed Central

    Desai, Sanjay A.

    2014-01-01

    Malaria parasites increase erythrocyte permeability to diverse solutes including anions, some cations, and organic solutes, as characterized with several independent methods. Over the last decade, patch-clamp studies have determined that the permeability results from one or more ion channels on the infected erythrocyte host membrane. However, the biological role(s) served by these channels, if any, remain controversial. Recent studies implicate the plasmodial surface anion channel (PSAC) and a role in parasite nutrient acquisition. A debated alternative role in remodeling host ion composition for the benefit of the parasite appears to be nonessential. Because both channel activity and the associated clag3 genes are strictly conserved in malaria parasites, channel-mediated permeability is an attractive target for development of new therapies. PMID:24507014

  16. Concentration of erythrocyte-based magnetic carriers in the bloodstream

    SciTech Connect

    Danilov, Y.N.; Il'ina, M.B.; Makharudov, S.Y.; Orekhov, A.N.; Rodchenko, S.A.; Samokhin, G.P.

    1986-04-01

    The writers postulated that magnetic erythrocytes (ME), injected into the bloodstream, may be concentrated in an assigned region of the vascular bed with the aid of the field of a permanent magnet. To test this hypothesis, erythrocytes ''loaded'' with colloidal magnetite were used, and concentrated in experiments in vitro and in vivo. For the experiments in vivo ME were labeled with sodium pertechnetate (Na /SUP 99n/ TcO4) was estimated in ME with a Rackgamma 1270 counter by determining radioactivity in the ME suspension and in the supernatant. For the experiment in vivo, a midline laparotomy was performed on a dog weighing 14 kg under intravenous hexobarbital anesthesia. The distribution of ME was recorded by means of a gamma-camera.

  17. Piezo1 plays a role in erythrocyte volume homeostasis

    PubMed Central

    Faucherre, Adèle; Kissa, Karima; Nargeot, Joël; Mangoni, Matteo E.; Jopling, Chris

    2014-01-01

    Mechanosensitivity is an inherent property of virtually all cell types, allowing them to sense and respond to physical environmental stimuli. Stretch-activated ion channels represent a class of mechanosensitive proteins which allow cells to respond rapidly to changes in membrane tension; however their identity has remained elusive. The piezo genes have recently been identified as a family of stretch-activated mechanosensitive ion channels. We set out to determine the role of piezo1 during zebrafish development. Here we report that morpholino-mediated knockdown of piezo1 impairs erythrocyte survival without affecting hematopoiesis or differentiation. Our results demonstrate that piezo1 is involved in erythrocyte volume homeostasis, disruption of which results in swelling/lysis of red blood cells and consequent anemia. PMID:23872304

  18. Variations of the Platelet Count in Disease

    PubMed Central

    Marchasin, Sidney; Wallerstein, Ralph O.; Aggeler, Paul M.

    1964-01-01

    Platelet counts were obtained in 675 patients with different hematological and other medical disorders. An indirect venous blood dry slide method which gave a normal range of 200 to 400 × 103 per cu mm was used. Platelet counts varied considerably in disease: In 20 patients, exclusive of myeloproliferative disorders, platelet counts in excess of 1,000 × 103 per cu mm were observed; in 20 patients, exclusive of leukemia and megaloblastic anemia, platelet counts were below 100 × 103 per cu mm. In general, platelet counts varied with the leukocyte count, but not with the degree of anemia. PMID:14180504

  19. α- and β-Monosaccharide transport in human erythrocytes

    PubMed Central

    Leitch, Jeffry M.; Carruthers, Anthony

    2009-01-01

    Equilibrative sugar uptake in human erythrocytes is characterized by a rapid phase, which equilibrates 66% of the cell water, and by a slow phase, which equilibrates 33% of the cell water. This behavior has been attributed to the preferential transport of β-sugars by erythrocytes (Leitch JM, Carruthers A. Am J Physiol Cell Physiol 292: C974–C986, 2007). The present study tests this hypothesis. The anomer theory requires that the relative compartment sizes of rapid and slow transport phases are determined by the proportions of β- and α-sugar in aqueous solution. This is observed with d-glucose and 3-O-methylglucose but not with 2-deoxy-d-glucose and d-mannose. The anomer hypothesis predicts that the slow transport phase, which represents α-sugar transport, is eliminated when anomerization is accelerated to generate the more rapidly transported β-sugar. Exogenous, intracellular mutarotase accelerates anomerization but has no effect on transport. The anomer hypothesis requires that transport inhibitors inhibit rapid and slow transport phases equally. This is observed with the endofacial site inhibitor cytochalasin B but not with the exofacial site inhibitors maltose or phloretin, which inhibit only the rapid phase. Direct measurement of α- and β-sugar uptake demonstrates that erythrocytes transport α- and β-sugars with equal avidity. These findings refute the hypothesis that erythrocytes preferentially transport β-sugars. We demonstrate that biphasic 3-O-methylglucose equilibrium exchange kinetics refute the simple carrier hypothesis for protein-mediated sugar transport but are compatible with a fixed-site transport mechanism regulated by intracellular ATP and cell shape. PMID:18987250

  20. Triggering of Suicidal Erythrocyte Death by the Antibiotic Ionophore Nigericin.

    PubMed

    Bissinger, Rosi; Malik, Abaid; Bouguerra, Ghada; Zhou, Yuetao; Singh, Yogesh; Abbès, Salem; Lang, Florian

    2016-05-01

    The K(+),H(+) ionophore and antibiotic nigericin has been shown to trigger apoptosis and is thus considered for the treatment of malignancy. Cellular mechanisms involved include induction of oxidative stress, which is known to activate erythrocytic Ca(2+)-permeable unselective cation channels leading to Ca(2+) entry, increase in cytosolic Ca(2+) activity ([Ca(2+)]i) and subsequent stimulation of eryptosis, the suicidal erythrocyte death characterized by cell shrinkage and cell membrane scrambling with phosphatidylserine translocation to the erythrocyte surface. This study explored whether and how nigericin induces eryptosis. Phosphatidylserine exposure at the cell surface was estimated from annexin V binding, cell volume from forward scatter, [Ca(2+)]i from Fluo3 fluorescence, pHi from BCECF fluorescence, ceramide abundance utilizing antibodies and reactive oxygen species (ROS) formation from DCFDA-dependent fluorescence. A 48-hr exposure of human erythrocytes to nigericin significantly increased the percentage of annexin-V-binding cells (0.1-10 nM), significantly decreased forward scatter (0.1-1 nM), significantly decreased cytosolic pH (0.1-1 nM) and significantly increased Fluo3 fluorescence (0.1-10 nM). Nigericin (1 nM) slightly, but significantly, increased ROS, but did not significantly modify ceramide abundance. The effect of nigericin on annexin V binding was significantly blunted, but not abolished by removal of extracellular Ca(2+). The nigericin-induced increase in [Ca(2+)]i and annexin V binding was again significantly blunted but not abolished by the Na(+)/H(+) exchanger inhibitor cariporide (10 μM). Nigericin triggers eryptosis, an effect paralleled by ROS formation, in part dependent on stimulation of Ca(2+) entry, and involving the cariporide-sensitive Na(+)/H(+) exchanger. © 2015 Nordic Association for the Publication of BCPT (former Nordic Pharmacological Society).

  1. Transport of lithium across the lamprey (Lampetra fluviatilis) erythrocyte membrane.

    PubMed

    Gusev, Gennadii P; Agalakova, Natalia I; Ivanova, Tatiana I

    2008-12-01

    Lithium, capable of replacing Na+ in various membrane transport processes, was used to investigate Na+ transport pathways across the lamprey erythrocytes membrane. The values of Li+ influxes have ranged from 8 to 24 mmol/l cells/h. Intracellular accumulation of Li+ was associated with loss of cellular Na+, the value of which was less than the value of Li+ influx. Both Li+ influx and Na+ efflux were partially inhibited by amiloride. The amiloride-sensitive Li+ influx was considerably stimulated by hyperosmotic cell shrinkage. The treatment of lamprey erythrocytes with blockers of protein phosphatases (fluoride and cantharidin) also resulted in a considerable increase in Li+ accumulation within the cells. No significant difference was observed between the values of Li+ and Na+ (22Na) influxes measured in red cells incubated simultaneously in isotonic LiCl and NaCl media (9.2 +/- 2.1 and 7.8 +/- 1.3 mmol/l cells/h, respectively). In hypo- and hypertonic media, however, the rate of Na+ influx in lamprey erythrocytes was approximately twice higher as compared to the rate of Li+ influx, what was determined by the difference in the amiloride-sensitive components. In acidified lamprey erythrocytes (intracellular pH 6.0) Li+ and Na+ influxes were considerably increased due to activation of amiloride-sensitive Na+/H+ (Li+/H+) exchange mechanism, although the activity of Na+/H+ exchange was much greater than that of Li+/H+ exchange. The data obtained confirm the hypothesis on the presence of two amiloride-sensitive systems of Na+ transport in the lamprey red blood cells.

  2. Repression and reactivation of lithium efflux from erythrocytes.

    PubMed

    Goodnick, P J; Meltzer, H L; Dunner, D L; Fieve, R R

    1979-10-01

    Efflux of lithium from human erythrocytes was studied in patients before, during, and after discontinuation of administration of lithium carbonate. Onset of lithium-induced repression of efflux took approximately 10 days and was significantly shorter in patients who had had lithium therapy previously. Reactivation took a longer period of time--approximately 2 week--and was found to be related to duration of lithium therapy. Theoretical pathways of lithium flow through membranes are discussed.

  3. Influence of calcium blockers on the SPR of erythrocytes

    NASA Astrophysics Data System (ADS)

    Shynkarenko, Olena V.; Tril, Orest; Wojnarowska, Renata; Prohorenko, Sergiy; Shergii, E. M.

    2016-12-01

    One of the promising areas of research is the impact of calcium channel blockers (CB) of biological fluids. This paper shows that the CB impact on a biological fluid can be efficiently combine with the surface plasmon resonance (SPR). It is shown that the addition of CB at the SPR measurements affect the stability of membranes and acts differently on the kinetics of erythrocytes ligament in the different groups of people.

  4. Stability of erythrocyte suspensions layered on stationary and flowing liquids

    NASA Technical Reports Server (NTRS)

    Omenyi, S. N.; Rhodes, P. H.; Snyder, R. S.

    1982-01-01

    The apparent stability of erythrocyte suspensions layered on stationary and flowing Ficoll solutions was studied considering the effects of particle concentration, type and size, and the different flow rates of the particle suspensions and chamber liquid. The data from the flowing system were empirically fitted and, when extrapolated to zero chamber liquid flow rate, gave values comparable to the data from the stationary system, thus confirming the validity of the data and our approach to obtain that data.

  5. Diabetic foot disease is associated with reduced erythrocyte deformability.

    PubMed

    Cahn, Avivit; Livshits, Leonid; Srulevich, Ariel; Raz, Itamar; Yedgar, Shaul; Barshtein, Gregory

    2016-08-01

    The pathogenesis of diabetic foot disease is multifactorial and encompasses microvascular and macrovascular pathologies. Abnormal blood rheology may also play a part in its development. Using a cell flow analyser (CFA), we examined the association between erythrocyte deformability and diabetic foot disease. Erythrocytes from diabetic patients with no known microvascular complications (n = 11) and patients suffering from a diabetic foot ulcer (n = 11) were isolated and their average elongation ratio (ER) as well as the ER distribution curve were measured. Average ER was decreased in the diabetic foot patients compared with the patients with diabetes and no complications (1·64 ± 0·07 versus 1·71 ± 0·1; P = 0·036). A significant rise in the percentage of minimally deformable red blood cells RBCs in diabetic foot patients compared with the patients with no complications was observed (37·89% ± 8·12% versus 30·61% ± 10·17%; P = 0·039) accompanied by a significant decrease in the percentage of highly deformable RBCs (12·47% ± 4·43% versus 17·49% ± 8·17% P = 0·046). Reduced erythrocyte deformability may slow capillary flow in the microvasculature and prolong wound healing in diabetic foot patients. Conversely, it may be the low-grade inflammatory state imposed by diabetic foot disease that reduces erythrocyte deformability. Further study of the rheological changes associated with diabetic foot disease may enhance our understanding of its pathogenesis and aid in the study of novel therapeutic approaches.

  6. Erythrocyte selenium and breast cancer risk: brief reports

    SciTech Connect

    Meyer, F.; Verreault, R.

    1987-05-01

    Animal experiments and ecologic studies suggest that low dietary selenium intake is associated with an increased risk of some types of cancer. However, studies assessing the relation of indicators of selenium intake in subjects to site-specific cancer incidence are few. This paper reports the results of a case-control study of erythrocyte selenium in relation to breast carcinoma in premenopausal women. 1 figure, 1 table.

  7. SWELLING OF ERYTHROCYTES IN SOLUTIONS OF AMMONIUM SALTS

    PubMed Central

    Schiödt, E.

    1933-01-01

    Two rather simple equations have been derived, which make it possible to express in a single number the result of a series of determinations of the volume of erythrocytes swelling in solutions of ammonium salts. In all experiments made with several combinations of different concentrations of permeating and non-permeating salts, the curves calculated from the equations have covered the points found by experiment. PMID:19872754

  8. Cryopreserved erythrocytes in clinical laboratory hemagglutination and hemagglutination inhibition tests.

    PubMed Central

    Maes, R K; Hayes, M M; Newman, J P

    1985-01-01

    Various concentrations of pig, guinea pig, chicken, and turkey erythrocytes (RBCs) were cryopreserved and compared with fresh RBCs in hemagglutination and hemagglutination inhibition tests. Hemagglutination and hemagglutination inhibition titers obtained with cryopreserved or fresh RBCs from the same donor differed by not more than one twofold dilution. RBCs cryopreserved at a concentration of less than or equal to 12.5% gave the highest percent recovery upon thawing. PMID:4077971

  9. Erythrocyte Antioxidant Defenses Against Cigarette Smoking in Ischemic Heart Disease

    PubMed Central

    Basalingappa, Doddamani R; Uppala, Satyanarayana; Mitta, Geeta

    2015-01-01

    Background Cigarette smoke promotes atherogenesis by producing oxygen-derived free radicals. Aim The present study was conducted to determine the effect of cigarette smoking on lipid peroxidation and erythrocyte antioxidant status in ischemic heart disease (IHD). Materials and Methods A total of 327 male subjects were enrolled for this study, divided into two groups consisting of 200 patients, who were consecutively admitted for IHD in the intensive cardiac care unit (ICCU) of a Government Hospital and 127 age matched male healthy subjects. Both the groups were subsequently categorised into smokers and non smokers sub groups depending upon the smoking history {>/= 20 pack years of smoking; (20 cigarettes per day for one year constitutes one pack year)}. All 327 subjects were investigated for lipid profile, malondialdehyde (MDA) levels and the antioxidant enzymes catalase (CAT), superoxide dismutase (SOD) and glutathione peroxidase (GPX). Statistical Analysis The differences in the parameters between the groups were tested for significance by one way ANOVA using the SPSS software version 19. A p-value of < 0.001 was considered to be significant statistically. Multiple comparisons were made between all the four groups by Post Hoc Tukey test. Results There was highly significant difference (p<0.001) observed in GPX activity, in comparison to CAT and SOD (p=0.032, p=0.009) between smokers vs non smokers in control group as well as patient group. The plasma MDA levels were found to be increased significantly (p<0.001) in IHD patients, who smoked as compared to those who did not. Conclusion Chronic smoking enhances erythrocyte lipid peroxidation in IHD patients with concomitant failure of both plasma and erythrocyte antioxidant defense mechanisms. Along with conventional lipid markers and plasma MDA levels, the erythrocyte GPX activity was observed to be a better marker of oxidative stress, in chronic smokers, who are at risk of developing IHD. PMID:26266112

  10. Acoustic measurement of compressibility and thermal expansion coefficient of erythrocytes

    NASA Astrophysics Data System (ADS)

    Toubal, Malika; Asmani, Mohamed; Radziszewski, Edouard; Nongaillard, Bertrand

    1999-05-01

    Mechanical properties of human erythrocytes, namely adiabatic compressibility and thermal expansion coefficient, have been determined using a classical ultrasound velocity and attenuation burst transmission technique. The theoretical model concerns the corpuscular part of the elastic wave propagating in a suspension of viscous particles of small size compared with the wavelength. The thermal wave contribution was taken into account. Normal and stiffened red blood cells were suspended in saline of different NaCl concentration.

  11. Assessment of chronic administration of Aloe vera gel on haematology, plasma biochemistry, lipid profiles and erythrocyte osmotic resistance in Wistar rats.

    PubMed

    Iji, O T; Oyagbemi, A A; Azeez, O I

    2010-11-28

    The study was designed to investigate the effects of chronic administration of Aloe vera gel extract on markers of hepatic damage, lipid profiles and erythrocyte osmotic fragility using the Wistar rats. Forty male Wistar rats divided into four groups of ten rats per group were used in the study. Group I which served as the control received 0.9% physiological saline while those in groups II, III and IV received Aloe vera gel (100, 250 and 500mg/kg), respectively, for four weeks. There was significant increase in the haemoglobin concentration while the PCV, RBC count, MCH and MCHC though showed some marginal increases but the increases were not significant in all the treated rats. No significant change was also observed in the erythrocyte osmotic fragility. However, there were significant reductions in plasma ALT, AST and ALP levels in animals that received the gel compared with the control while the plasma albumin and total protein values were higher than those of the control. All the animals that received the gel also showed significant reduction in plasma total cholesterol, triglycerides and LDL-cholesterol ratio compared with the control. In a similar manner, those animals that were administered with 500mg/kg gel had significantly higher HDL-cholesterol ratio than those of the control. This study showed that, chronic administration of Aloe vera gel extract had no significant effects on the haematological parameters of the rats and did not affect erythrocyte osmotic resistance. It however showed some cholesterol lowering action.

  12. Simultaneous measurements of the accumulation of isotope-labelled protein and erythrocytes in skin reactions of allergic inflammation in the guinea-pig

    PubMed Central

    Morley, J.; Williams, T. J.; Slater, A. J.; Cubitt, D.; Dumonde, D. C.

    1972-01-01

    This paper describes a method for simultaneous measurement of the accumulation of plasma protein and erythrocytes in skin reactions of hypersensitivity to bovine γ-globulin (BGG) and tuberculin PPD in the guinea-pig. The procedure consists in giving 125I-labelled plasma albumin and 51Cr-labelled autologous erythrocytes together by intravenous injection into guinea-pigs bearing skin lesions of allergic inflammation at different times and for different periods during the development of the skin reactions. Isotope accumulation in excised skin reactions is measured by scintillation counting at different stages during the evolution of hypersensitivity responses. Skin reactions of combined anaphylactic and Arthus hypersensitivity to BGG were characterized by pronounced increased vascular permeability principally in the first hour. In established 24-hour hypersensitivity reactions to both antigens (BGG and PPD) there was continuing accumulation of both plasma albumin and erythrocytes. During the development of the tuberculin reaction, an intermediate phase of isotope accumulation occurred between 6 and 9 hours after skin testing; serum transfer studies showed that this intermediate peak was not attributable to circulating antibody alone. These isotope tracer techniques were also applied to study vascular permeability in systemically transferred reactions of delayed hypersensitivity and in the vascular response to the intradermal injection of an inflammatory factor generated by antigen-activation of sensitized lymphocytes. It was concluded that isotope tracing provided objective and sensitive methods for analysing microvascular responses in allergic inflammation. PMID:4262654

  13. Effect of bacterial peptidoglycan on erythrocyte death and adhesion to endothelial cells.

    PubMed

    Abed, Majed; Towhid, Syeda T; Pakladok, Tatsiana; Alesutan, Ioana; Götz, Friedrich; Gulbins, Erich; Lang, Florian

    2013-05-01

    Peptidoglycans, bacterial wall components, have previously been shown to trigger eryptosis, the suicidal erythrocyte death, characterized by cell shrinkage and cell membrane scrambling with phosphatidylserine exposure at the cell surface. Phosphatidylserine exposing erythrocytes adhere to the vascular wall at least partially by interaction of erythrocytic phosphatidylserine with endothelial CXC chemokine ligand 16 (CXCL16). The present study explored whether peptidoglycan exposure fosters the adhesion of erythrocytes to human umbilical vein endothelial cells (HUVEC). To this end, HUVEC were treated for 48 h with peptidoglycan (10 μg/ml) and CXCL16 abundance determined by confocal microscopy and FACS analysis. Moreover, human erythrocytes were exposed for 48 h to peptidoglycan (10 μg/ml) and phosphatidylserine exposure estimated from binding of fluorescent annexin-V, cell volume from forward scatter in FACS analysis and erythrocyte adhesion to human umbilical vein endothelial cells (HUVEC) from trapping of labeled erythrocytes in a flow chamber. As a result, bacterial peptidoglycan exposure was followed by increased CXCL16 expression in HUVEC as well as erythrocyte shrinkage, phosphatidylserine exposure and adhesion to HUVEC under flow conditions at arterial shear rates. The adhesion was significantly attenuated but not abrogated in the presence of either, erythrocyte phosphatidylserine-coating annexin-V (5 μl/ml) or CXCL16 neutralizing antibody directed against endothelial CXCL16 (4 μg/ml). In conclusion, exposure to peptidoglycan increases endothelial CXCL16 expression and leads to eryptosis followed by phosphatidylserine- and CXCL16-mediated adhesion of eryptotic erythrocytes to vascular endothelial cells.

  14. Human erythrocytes inhibit complement-mediated solubilization of immune complexes by human serum

    SciTech Connect

    Dorval, B.L.

    1987-01-01

    The aim of this study was to develop an autologus human system to evaluate the effects of human erythrocytes on solubilization of immune complex precipitates (IC) by human serum. Incubation of IC with fresh human serum or guinea pig serum resulted in solubilization of IC. When packed erythrocytes were added to human serum or guinea pig serum binding of IC to the erythrocyte occurred and IC solubilization was inhibited significantly (p <.025). Sheep erythrocytes did not bind IC or inhibit IC solubilization. To evaluate the role of human erythrocyte complement receptor (CR1) on these findings, human erythrocytes were treated with trypsin or anti-CR1 antibodies. Both treatments abrogated IC binding to human erythrocytes but did not affect the ability of the human erythrocyte to inhibit IC solubilization. Radioimmunoassay was used to measure C3, C4 and C5 activation in human serum after incubation with IC, human erythrocytes, human erythrocytes plus IC, whole blood or in whole blood plus IC.

  15. Biochemically altered human erythrocytes as a carrier for targeted delivery of primaquine: an in vitro study.

    PubMed

    Alanazi, Fars K; Harisa, Gamal El-Din I; Maqboul, Ahmad; Abdel-Hamid, Magdi; Neau, Steven H; Alsarra, Ibrahim A

    2011-04-01

    The aim of this study was to investigate human erythrocytes as a carrier for targeted drug delivery of primaquine (PQ). The process of PQ loading in human erythrocytes, as well as the effect of PQ loading on the oxidative status of erythrocytes, was also studied. At PQ concentrations of 2, 4, 6, and 8 mg/mL and an incubation time of 2 h, the ratios of the concentrations of PQ entrapped in erythrocytes to that in the incubation medium were 0.515, 0.688, 0.697 and 0.788, respectively. The maximal decline of erythrocyte reduced glutathione content was observed at 8 mg/mL of PQ compared with native erythrocytes p < 0.001. In contrast, malondialdehyde and protein carbonyl were significantly increased in cells loaded with PQ (p < 0.001). Furthermore, osmotic fragility of PQ carrier erythrocytes was increased in comparison with unloaded cells. Electron microscopy revealed spherocyte formation with PQ carrier erythrocytes. PQ-loaded cells showed sustained drug release over a 48 h period. Erythrocytes were loaded with PQ successfully, but there were some biochemical as well as physiological changes that resulted from the effect of PQ on the oxidative status of drug-loaded erythrocytes. These changes may result in favorable targeting of PQ-loaded cells to reticulo-endothelial organs. The relative impact of these changes remains to be explored in ongoing animal studies.

  16. Erythrocyte membrane stability to hydrogen peroxide is decreased in Alzheimer disease.

    PubMed

    Gilca, Marilena; Lixandru, Daniela; Gaman, Laura; Vîrgolici, Bogdana; Atanasiu, Valeriu; Stoian, Irina

    2014-01-01

    The brain and erythrocytes have similar susceptibility toward free radicals. Therefore, erythrocyte abnormalities might indicate the progression of the oxidative damage in Alzheimer disease (AD). The aim of this study was to investigate erythrocyte membrane stability and plasma antioxidant status in AD. Fasting blood samples (from 17 patients with AD and 14 healthy controls) were obtained and erythrocyte membrane stability against hydrogen peroxide and 2,2'-azobis-(2-amidinopropane) dihydrochloride (AAPH), serum Trolox equivalent antioxidant capacity (TEAC), residual antioxidant activity or gap (GAP), erythrocyte catalase activity (CAT), erythrocyte superoxide dismutase (SOD) activity, erythrocyte nonproteic thiols, and total plasma thiols were determined. A significant decrease in erythrocyte membrane stability to hydrogen peroxide was found in AD patients when compared with controls (P<0.05). On the contrary, CAT activity (P<0.0001) and total plasma thiols (P<0.05) were increased in patients with AD compared with controls. Our results indicate that the most satisfactory measurement of the oxidative stress level in the blood of patients with AD is the erythrocyte membrane stability to hydrogen peroxide. Reduced erythrocyte membrane stability may be further evaluated as a potential peripheral marker for oxidative damage in AD.

  17. Bilayer/cytoskeleton interactions in lipid-symmetric erythrocytes assessed by a photoactivable phospholipid analogue

    SciTech Connect

    Pradhan, D.; Schlegel, R.A. ); Williamson, P. )

    1991-08-06

    Two mechanisms have been proposed for maintenance of transbilayer phospholipid asymmetry in the erythrocyte plasma membrane, one involving specific interactions between the aminophospholipids of the inner leaflet of the bilayer and the cytoskeleton, particularly spectrin, and the other involving the aminophospholipid translocase. If the former mechanism is correct, then erythrocytes which have lost their asymmetric distribution of phospholipids should display altered bilayer/cytoskeleton interactions. To test this possibility, normal erythrocytes, erythrocytes from patients with chronic myelogenous leukemia or sickle disease, and lipid-symmetric and -asymmetric erythrocyte ghosts were labeled with the radioactive photoactivable analogue of phosphatidylethanolamine, 2-(2-azido-4-nitrobenzoyl)-1-acyl-sn-glycero-3-phospho({sup 14}C) ethanolamine (({sup 14}C)AzPE), previously shown to label cytoskeletal proteins from the bilayer. The labeling pattern of cytoskeletal proteins in pathologic erythrocytes and lipid-asymmetric erythrocyte ghosts was indistinguishable from normal erythrocytes, indicating that the probe detects no differences in bilayer/cytoskeleton interactions in these cells. In contrast, in lipid-symmetric erythrocyte ghosts, labeling of bands 4.1 and 4.2 and actin, and to a lesser extent ankyrin, by ({sup 14}C)AzPE was considerably reduced. Significantly, however, labeling of spectrin was unaltered in the lipid-symmetric cells. These results do not support a model in which spectrin is involved in the maintenance of an asymmetric distribution of phospholipids in erythrocytes.

  18. [Calcium-transporting system of erythrocytes in psoriasis].

    PubMed

    Mavrov, I I; Goncharenko, M S; Petruniaka, V V; Ereshchenko, E A; Kondakova, A K; Stepaniuk, L V

    1990-01-01

    The activity of Ca-ATPase and permeability of erythrocyte membrane for calcium in patients with psoriasis were studied with the aim to reveal disturbances in the calcium membrane transport under psoriasis. In the presence of endogenic activators the mean values of the maximal Ca-ATPase activity of erythrocyte membranes in patients with psoriasis and in healthy people have no essential differences and make up 264 +/- 12 and 244 +/- 10 mumol P/1 cells per 1 min, respectively. The rate of 45Ca accumulation in erythrocytes under inhibition of Ca-ATPase in patients suffering from psoriasis is by 64% higher than in healthy people. The data obtained along with the previously revealed changes in the calcium metabolism in patients with psoriasis make it possible to suppose the presence of the system disturbance of the calcium membrane transport, in particular an increase in the plasma membrane permeability for cells of different types. Such a disturbance may distort a regulatory (messenger) function of calcium ions in the processes of proliferation, differentiation, functional activity and death of different cell types.

  19. Erythrocyte sodium/potassium ATPase activity in severe preeclampsia.

    PubMed

    Adair, C D; Haupert, G T; Koh, H P; Wang, Y; Veille, J-C; Buckalew, V

    2009-04-01

    Elevated blood levels of endogenous digitalis-like factors (EDLF) may decrease erythrocyte sodium pump activity in preeclampsia. As the highest EDLF levels might be expected in severe preeclampsia, we investigated sodium pump activity in that group of patients. Erythrocyte sodium pump activity was determined by (86)Rubidium uptake (in nM per hour per 10(6) cells) in women with severe preeclampsia and those with normal pregnancies, matched for gestational age, and in healthy nonpregnant women (n=12 in each group). Differences between groups were analyzed by a two-sided Student t-test. Sodium pump activity was significantly increased in normotensive pregnancies as compared with normotensive non-pregnant women (81.4+/-8.4 vs 61.1+/-7.4, mean+/-s.d., p<0.05), and was decreased 43% in severe preeclamptic pregnancies as compared with normotensive pregnancies (46.4+/-14.1 vs 81.4+/-8.4, p<0.05). Severe preeclampsia is associated with significantly lower erythrocyte sodium pump activity than normotensive pregnancy. These data suggest that plasma levels of a biologically active EDLF are elevated in patients with severe preeclampsia.

  20. Drug-loaded erythrocytes: on the road toward marketing approval

    PubMed Central

    Bourgeaux, Vanessa; Lanao, José M; Bax, Bridget E; Godfrin, Yann

    2016-01-01

    Erythrocyte drug encapsulation is one of the most promising therapeutic alternative approaches for the administration of toxic or rapidly cleared drugs. Drug-loaded erythrocytes can operate through one of the three main mechanisms of action: extension of circulation half-life (bioreactor), slow drug release, or specific organ targeting. Although the clinical development of erythrocyte carriers is confronted with regulatory and development process challenges, industrial development is expanding. The manufacture of this type of product can be either centralized or bedside based, and different procedures are employed for the encapsulation of therapeutic agents. The major challenges for successful industrialization include production scalability, process validation, and quality control of the released therapeutic agents. Advantages and drawbacks of the different manufacturing processes as well as success key points of clinical development are discussed. Several entrapment technologies based on osmotic methods have been industrialized. Companies have already achieved many of the critical clinical stages, thus providing the opportunity in the future to cover a wide range of diseases for which effective therapies are not currently available. PMID:26929599