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Sample records for agglutinated rabbit erythrocytes

  1. SUPPRESSION OF BLOOD GROUP AGGLUTINABILITY OF HUMAN ERYTHROCYTES BY CERTAIN BACTERIAL POLYSACCHARIDES

    PubMed Central

    Ceppellini, Ruggero; Landy, Maurice

    1963-01-01

    Erythrocytes coated with bacterial capsular polysaccharides, notably the Vi antigen, were no longer agglutinated by antibodies directed against the various antigens native to the red cell surface. These effects could not be attributed to prevention of antibody uptake even though in some systems the uptake of antibody was diminished. In fact, agglutination by Rh-incomplete antibody was brought back to the original titer only after the sensitized Vi-coated cells had been subjected to ten alternating exposures to globulin and antiglobulin. Hemagglutination by Newcastle, mumps, and influenza viruses was also suppressed. Erythrocytes coated with Vi polysaccharide assumed the distinctive physicochemical attributes of this acidic polymer which results in a stabilization of the erythrocyte suspension as manifested by increased electrophoretic mobility and a striking decrease in the rate of sedimentation. Among the possible models for explaining the nature of the Vi effect on immune agglutination, the data favor interference with lattice formation. PMID:14019651

  2. 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.

  3. Aeromonas hydrophila typing scheme based on patterns of agglutination with erythrocytes and yeast cells.

    PubMed Central

    Adams, D; Atkinson, H M; Woods, W H

    1983-01-01

    An agglutination typing scheme has been developed for strains of Aeromonas hydrophila. Primary agglutination typing is based on testing agar-grown A. hydrophila cells with human, horse, rat, and guinea pig erythrocytes and Saccharomyces cerevisiae cells. Further subdivision of primary groups is based firstly on whether yeast cell agglutination is inhibited by a D-mannose polymer, yeast mannan, and secondly on patterns of inhibition of hemagglutination by yeast mannan and the monomeric sugars L-fucose, D-galactose, and D-mannose. A total of 320 isolates were tested, and these were divisible into 39 distinct types on the basis of this scheme. Application of this typing scheme in the future to isolates of A. hydrophila known to be associated with human infection may enable correlations to be made between particular agglutination types and human pathogenicity. PMID:6841579

  4. 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.

  5. Agglutination of human O erythrocytes by influenza A(H1N1) viruses freshly isolated from patients.

    PubMed

    Murakami, T; Haruki, K; Seto, Y; Kimura, T; Minoshiro, S; Shibe, K

    1991-04-01

    The hemagglutinin titers of 10 influenza A (H1N1) viruses were examined using the erythrocytes of several species. Human O erythrocytes showed the highest agglutination titer to the viruses, whereas chicken erythrocytes showed a low titer. These findings were noted for at least 10 passages by serial dilutions of the viruses in Madin-Darby canine kidney (MDCK) cells. All influenza A(H1N1) viruses, plaque-cloned directly from throat-washing specimens of patients, also agglutinated human O but not chicken erythrocytes. The results of a hemadsorption test indicated that chicken erythrocytes possess less affinity to MDCK cells infected with the A/Osaka City/2/88(H1N1) stain than to those infected with the A/Yamagata/120/86(H1N1) strain which is used as an inactivated influenza vaccine in Japan. However, there were no significant differences between the A/Osaka City/2/88 and the A/Yamagata/120/86 strains in the hemagglutination inhibition test. Since human O erythrocytes have high agglutination activity to influenza A(H1N1) and also to A(H3N2) and B viruses in MDCK cells, these erythrocytes may be useful for the serological diagnosis of influenza.

  6. Agglutination of human erythrocytes by the interaction of Zn(2+)ion with histidine-651 on the extracellular domain of band 3.

    PubMed

    Kiyotake, Kento; Ochiai, Hideharu; Yamaguchi, Takeo

    2016-05-01

    Clustering of band 3, chloride/bicarbonate exchanger, has been reported in Zn(2+)-treated human erythrocytes. However, the agglutination of human erythrocytes is also induced by the interaction of Zn(2+)ion with histidine on band 3. Identification of histidine that interacts with Zn(2+)ion remains to be determined. The Zn(2+)-induced agglutination of human erythrocytes was unaffected by chymotrypsin cleavage of the small loop region containing His-547 in the extracellular domain of band 3. On the other hand, papain digestion of the large loop region containing His-651 in band 3 inhibited such Zn(2+)-induced agglutination. Moreover, Zn(2+)-induced erythrocyte agglutination was inhibited by the peptide (ARGWVIHPLG) containing His-651, but not by the peptide such as ARGWVIRPLG, which His-651 was substituted by arginine. Among 10 kinds of animal erythrocytes tested, interestingly, no agglutination by Zn(2+)ions was observed in cow cells only that the forth amino acid in the upstream from His-669 on the large loop of cow band 3 is aspartate (Asp-665) instead of glycine. As expected, the agglutination of human erythrocytes by Zn(2+) ions was inhibited in the presence of aspartate. These data indicate that the interaction of Zn(2+) ion with His-651 residue of band 3 plays an important role in the Zn(2+)-induced agglutination of human erythrocytes.

  7. Study of polycation effects on erythrocyte agglutination mediated by anti-glycophorins using microscopic image digital analysis

    NASA Astrophysics Data System (ADS)

    Riquelme, B.; Dumas, D.; Relancio, F.; Fontana, A.; Alessi, A.; Foresto, P.; Grandfils, C.; Stoltz, J.; Valverde, J.

    2006-04-01

    The aim of this work was to study synthetic polycation effects on erythrocyte agglutination mediated by anti-glycophorin using image digital analysis. Polycations are oligomers or polymers of natural or synthetic origin, which bear a great number of positive charges at pH 7.4. Several of these polycations are nowadays used in clinic for human and veterinary purposes. New applications of polycations to the development of new drug delivery systems are investigated, in order to promote the drug absorption through the gastro-intestinal and blood brain barriers. However, up to now, there are no clear relationships between macromolecular features of polycations (molecular weight, mean charge density, charge repartition, etc.) and their interactions with blood elements (which bear superficial negative charges). The interaction on the red blood cell membrane with synthetic polycations having well-controlled macromolecular features and functionalized with pendent polyethylene glycol segments was investigated. The alterations over stationary and dynamic viscoelastic properties of erythrocyte membranes were analyzed through laser diffractometry. Image digital analysis was used to study erythrocyte agglutination mediated by anti-glycophorin. Results show different reactivities of the polycations on the erythrocyte membrane. These findings could provide more information about the mechanisms of polycation interaction on erythrocyte membranes. We consider that this work could provide useful tools to understand and improve the haemocompatibility of polycations and enlarge their potential in clinic.

  8. Application of a spectrally filtered probing light beam and RGB decomposition of microphotographs for flow registration of ultrasonically enhanced agglutination of erythrocytes

    NASA Astrophysics Data System (ADS)

    Doubrovski, V. A.; Ganilova, Yu. A.; Zabenkov, I. V.

    2013-08-01

    We propose a development of the flow microscopy method to increase the resolving power upon registration of erythrocyte agglutination. We experimentally show that the action of a ultrasonic standing wave on an agglutinating mixture blood-serum leads to the formation of so large erythrocytic immune complexes that it seems possible to propose a new two-wave optical method of registration of the process of erythrocyte agglutination using the RGB decomposition of microphotographs of the flow of the mixture under study. This approach increases the reliability of registration of erythrocyte agglutination and, consequently, increases the reliability of blood typing. Our results can be used in the development of instruments for automatic human blood typing.

  9. Spectral dependence of resolving power of optical method of detection of ultrasonically enhanced agglutination of human blood erythrocytes

    NASA Astrophysics Data System (ADS)

    Doubrovski, V. A.; Dvoretski, K. N.; Dolmashkin, A. A.

    2010-08-01

    The spectral dependence of the resolving power of an acoustooptic method of monitoring agglutination of human blood erythrocytes is studied theoretically and experimentally. It is shown that, in principle, the resolving power of this method can be increased by several dozen times. The results of the work can be used to create instruments for determining the human blood type in the AB0 system and in the Rhesus system.

  10. [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.

  11. Diagnosis of myocardial infarction based on lectin-induced erythrocyte agglutination: a feasibility study

    NASA Astrophysics Data System (ADS)

    Bocsi, József; Nieschke, Kathleen; Mittag, Anja; Reichert, Thomas; Laffers, Wiebke; Marecka, Monika; Pierzchalski, Arkadiusz; Piltz, Joachim; Esche, Hans-Jürgen; Wolf, Günther; Dähnert, Ingo; Baumgartner, Adolf; Tarnok, Attila

    2014-03-01

    Myocardial infarction (MI) is an acute life-threatening disease with a high incidence worldwide. Aim of this study was to test lectin-carbohydrate binding-induced red blood cell (RBC) agglutination as an innovative tool for fast, precise and cost effective diagnosis of MI. Five lectins (Ricinus communis agglutinin (RCA), Phaseolus vulgaris erythroagglutinin (PHA), Datura stramonium agglutinin (DSA), Artocarpus agglutinin (ArA), Triticum agglutinin (TA)) were tested for ability to differentiate between agglutination characteristics in patients with MI (n = 101) or angina pectoris without MI (AP) (n = 34) and healthy volunteers (HV) as control (n =68) . RBC agglutination was analyzed by light absorbance of a stirred RBC suspension in the green to red light spectrum in an agglutimeter (amtec, Leipzig, Germany) for 15 min after lectin addition. Mean cell count in aggregates was estimated from light absorbance by a mathematical model. Each lectin induced RBC agglutination. RCA led to the strongest RBC agglutination (~500 RBCs/aggregate), while the others induced substantially slower agglutination and lead to smaller aggregate sizes (5-150 RBCs/aggregate). For all analyzed lectins the lectin-induced RBC agglutination of MI or AP patients was generally higher than for HV. However, only PHA induced agglutination that clearly distinguished MI from HV. Variance analysis showed that aggregate size after 15 min. agglutination induced by PHA was significantly higher in the MI group (143 RBCs/ aggregate) than in the HV (29 RBC-s/aggregate, p = 0.000). We hypothesize that pathological changes during MI induce modification of the carbohydrate composition on the RBC membrane and thus modify RBC agglutination. Occurrence of carbohydrate-lectin binding sites on RBC membranes provides evidence about MI. Due to significant difference in the rate of agglutination between MI > HV the differentiation between these groups is possible based on PHA-induced RBC-agglutination. This novel assay

  12. An acousto-optical method for registration of erythrocytes' agglutination reaction—sera color influence on the resolving power

    NASA Astrophysics Data System (ADS)

    Doubrovski, V. A.; Medvedeva, M. F.; Torbin, S. O.

    2016-01-01

    The absorption spectra of agglutinating sera were used to determine blood groups. It was shown experimentally that the sera color significantly affects the resolving power of the acousto-optical method of blood typing. In order to increase the resolving power of the method and produce an invariance of the method for sera color, we suggested introducing a probing light beam individually for different sera. The proposed technique not only improves the resolving power of the method, but also reduces the risk of false interpretation of the experimental results and, hence, error in determining the blood group of the sample. The latter is especially important for the typing of blood samples with weak agglutination of erythrocytes. This study can be used in the development of an instrument for instrumental human blood group typing based on the acousto-optical method.

  13. Viability of Stored Rabbit Erythrocytes Carrying Number C3c.

    DTIC Science & Technology

    1984-05-31

    A-A165 440 VIABILITY OF STORED RABBIT ERYTHROCYTES CARRYING NUBR V no C3C(U) MASSACHUSETTS UNIV MEDICAL SCHOOL MORCESTER UNCLRSSIFI 0 SZYMANSKI 31...Carrying No C3c 0 Annual and Final Report Irma 0. Szymanski, M.D. Ln CD May 31, 1984 I Supported by U.S. ARMY MEDICAL RESEARCH AND DEVELOPMENT COMMAND...Fort Detrick, Frederick, Maryland 21701-5012 Contract No. DAMD17-82-C-2150 .TICD ET University of Massachusetts Medical Center .EC, Worcester

  14. [The evaluation of different agglutination and adhesion tests with erythrocytic reagents in determining antibodies to the O and H antigens of Salmonella typhimurium in comparison with immunoenzyme analysis (IEA)].

    PubMed

    Karal'nik, B V; Denisova, T G

    1996-01-01

    The comparison of the effectiveness of EIA with that of a number of agglutination and adhesion tests with erythrocyte diagnostica in the determination of antibodies to different S.typhimurium antigens demonstrated higher sensitivity of EIA. The relative specificity of the determination of O- and H-antibodies in EIA and in agglutination and adhesion tests depended on the isotype of antibodies to be determined and the specificity of sensitins used in the production of immunoreagents.

  15. Erythrocyte migration and gap formation in rabbit blood clots in vitro.

    PubMed

    Ueki, T; Yazama, F; Horiuchi, T; Yamada, M

    2008-04-01

    Thrombolytic agents must be carried by the blood circulation to thrombi to exert their functions. Structural gaps exist between blood vessels and thrombi or in the area surrounding thrombi. Therefore, information about fundamental gap formation at thrombotic areas is critically important for thrombolytic therapy. We previously reported that t-PA accelerates the activities of bovine erythrocytes and hemoglobin (Hb) towards bovine plasminogen activation. Here, we examined gap generation by observing morphological changes during thrombolytic processes in rabbit blood clots deformation of erythrocytes from blood clots and Hb transfer from erythrocytes to serum in vitro. Rabbit venous blood samples (1 ml) were stored under sterile conditions in glass tubes at 37 degrees C for 2, 24, 48 h, 1, and 2 weeks. We examined clot diameter, erythrocyte diameter and number as well as Hb volume in the serum, as well as histological changes in the clots. The diameter of blood clots did not change until 2 weeks after sampling. Erythrocyte diameter decreased within 48 h and at 2 weeks after sampling at the clot surface (p < 0.001) and interior (p < 0.001). The number of erythrocytes in the serum started to increase starting from 24 h after sampling (p < 0.01). Serum Hb volume also gradually increased from 24 h until 2 weeks after sampling (p < 0.01). The erythrocyte envelope became disrupted and cytoplasm started to flow through pores into the serum at 24 h. The results indicated that blood clots are reduced due to clot retraction, erythrocyte dissociation and cytoplasm leakage without a distinct fibrinolytic reaction. These results indicated that gaps start to form between 2 and 24 h after blood clotting.

  16. 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.

  17. Evidence for coupling of Clostridium perfringens alpha-toxin-induced hemolysis to stimulated phosphatidic acid formation in rabbit erythrocytes.

    PubMed Central

    Sakurai, J; Ochi, S; Tanaka, H

    1993-01-01

    When rabbit erythrocytes were exposed to low concentrations of Clostridium perfringens alpha-toxin, hot-cold hemolysis was observed. The toxin induced production of phosphatidic acid (PA) in a dose-dependent manner when incubated with erythrocytes at 37 degrees C. When erythrocyte membranes were incubated with the toxin and [gamma-32P]ATP in the presence or absence of ethanol, [32P]PA formation was maximal within 30 s, then sharply decreased, and began again after 5 min of incubation. Ethanol had no effect on the early appearance (at approximately 5 min) of PA formation induced by the toxin but significantly inhibited formation of PA over 10 min of incubation. Treatment of erythrocyte membranes with alpha-toxin resulted in the biphasic formation of 1,2-diacylglycerol and PA as well as an increase of inositol-1,4,5-trisphosphate (IP3) and decrease of phosphatidylinositol-4,5-bisphosphate (PIP2) within 30 s. Neomycin inhibited the toxin-induced increase in turbidity of egg yolk suspensions but did not inhibit the toxin-induced hemolysis of intact erythrocytes. On the other hand, neomycin inhibited the toxin-induced hemolysis of saponin-treated erythrocytes. In addition, neomycin inhibited PA formation induced by the toxin in erythrocyte membranes. IP3 was released by incubation of PIP2 with erythrocyte membranes but not by incubation of PIP2 with the toxin. The toxin stimulated the membrane-induced release of IP3 from PIP2. These data suggest that the toxin-induced hemolysis is dependent on the action of phospholipase C in erythrocyte membranes. PMID:8395469

  18. In vitro antioxidant activities of resveratrol, cinnamaldehyde and their synergistic effect against cyadox-induced cytotoxicity in rabbit erythrocytes.

    PubMed

    Farag, Mayada Ragab; Alagawany, Mahmoud; Tufarelli, Vincenzo

    2017-04-01

    This study was conducted to explore the potential benefits of using cinnamaldehyde (CIN), resveratrol (RES) separately or in combination on cyadox (CYA)-induced alterations in isolated rabbit erythrocytes. Erythrocytes suspensions were partitioned into 7 groups (5 replicates/group), 1st kept as control treated with phosphate buffered saline (PBS) with dimethyl sulphoxide (DMSO); 2nd group was subjected to CYA (40 μg/ml), 3rd group was incubated with CIN (40 μM), 4th group was subjected to RES (40 μM), 5th group was co-exposed to CYA (40 μg/ml) and CIN (40 μM), 6th group was co exposed to CYA (40 μg/ml) and RES (40 μM), and 7th group was exposed to CYA in combination with both CIN and RES at the same indicated concentrations. The reaction mixtures of different groups were incubated at 37 °C for 3 h with gentle shaking every 15 minutes. Our results revealed that exposure to CYA caused a significant decrease (linear and quadratic) in superoxide dismutase (SOD) and catalase (CAT) activities and the contents of reduced glutathione (GSH) and glutathione transferase (GST). Incubation of erythrocytes with CYA increased GSSG content, GSSG/GSH ratio, malonaldehyde (MDA) and protein carbonyl (PrC) concentrations while it decreased the total protein (TP). CYA also lead to hemolysis and energy depletion of erythrocytes beside activation of caspase cascades, suggesting the pro-oxidant effect CYA that could be implicated in eryptosis. CIN and RES were able to inverse these hazardous effects of CYA. However, CIN was more effective than RES, their combination showed a positive synergistic effect in protecting the cells against oxidative injury caused by CYA.

  19. Volume regulatory potassium transport in rabbit and human sickle erythrocytes in vitro

    SciTech Connect

    Al-Rohil, N.S.

    1988-01-01

    One approach to the therapy of sickle cell anemia is to decrease the hemoglobin concentration by inducing a slight swelling of the cell to retard the rate of hemoglobin polymerization. We found that a prolonged incubation of rabbit or human SS red cell in hypotonic medium caused an inactivation of the inactivation of swelling-stimulated potassium transport. The inactivation may have important practical consequences for the therapy of sickle cell anemia. Large cytoskeleton-free vesicles were prepared in order to study the possible role of the spectrin-actin membrane skeleton in the swelling-stimulated and N-ethylmaleimide (NEM)-stimulated transport. NEM pretreatment stimulated {sup 86}Rb efflux in vesicles by a factor of 2.4 + 0.55 (mean {plus minus} S.D.). The NEM effect on {sup 86}Rb efflux was specific in that the {sup 22}Na efflux into a Na medium was not stimulated but actually inhibited. The {sup 86}Rb efflux from the vesicles was not stimulated by hypotonic media. This finding is consistent with a role of the membrane skeleton in the detection and/or transduction of the signal by which cell swelling activates the transport.

  20. Human blood group typing based on digital photographs of RBC agglutination process

    NASA Astrophysics Data System (ADS)

    Doubrovski, V. A.; Dolmashkin, A. A.

    2010-08-01

    A method for the monitoring of the human erythrocyte agglutination reaction in vitro, which is the basis for determining the human blood type (group), is proposed. The method is based on a statistical analysis of digital photographs of the agglutination process. It is experimentally shown that this analysis of photographs makes it possible to determine the probability that the agglutination reaction of erythrocytes of the studied specimen of blood with corresponding isohemagglutinating sera does occur. To increase the rate of the agglutination reaction of erythrocytes and to improve the sensitivity of the method of monitoring, the bioobject under study is subjected to the action of ultrasonic waves, as was previously proposed by the authors, and the result of the erythrocyte agglutination reaction is read optically. It is shown that, in principle, the method of statistical processing of digital photographs can be used to develop devices for automatic human blood typing in the AB0 and Rh systems.

  1. Agglutinating Activity and Structural Characterization of Scalarin, the Major Egg Protein of the Snail Pomacea scalaris (d’Orbigny, 1832)

    PubMed Central

    Ituarte, Santiago; Dreon, Marcos Sebastián; Ceolin, Marcelo; Heras, Horacio

    2012-01-01

    Apple snail perivitellins are emerging as ecologically important reproductive proteins. To elucidate if the protective functions of the egg proteins of Pomacea canaliculata (Caenogastropoda, Ampullariidae), involved in embryo defenses, are present in other Pomacea species we studied scalarin (PsSC), the major perivitellin of Pomacea scalaris. Using small angle X-ray scattering, fluorescence and absorption spectroscopy and biochemical methods, we analyzed PsSC structural stability, agglutinating activity, sugar specificity and protease resistance. PsSC aggluttinated rabbit, and, to a lesser extent, human B and A erythrocytes independently of divalent metals Ca2+ and Mg2+ were strongly inhibited by galactosamine and glucosamine. The protein was structurally stable between pH 2.0 to 10.0, though agglutination occurred only between pH 4.0 to 8.0 (maximum activity at pH 7.0). The agglutinating activity was conserved up to 60°C and completely lost above 80°C, in agreement with the structural thermal stability of the protein (up to 60°C). PsSC was able to withstand in vitro gastrointestinal digestion, and showed no trypsin inhibition activity. The presence of lectin activity has been reported in eggs of other Pomacea snails, but here we link for the first time, this activity to an apple snail multifunctional perivitellin. This novel role for a snail egg storage protein is different from closely related P.canaliculata defensive proteins. PMID:23185551

  2. Mannanoligosaccharide agglutination by Salmonella enterica strains isolated from carrier pigs

    PubMed Central

    Borowsky, Luciane; Corção, Gertrudes; Cardoso, Marisa

    2009-01-01

    Type-1 fimbriae are associated with most Salmonella enterica serovars and are an essential factor for host colonization. Mannanoligosaccharides (MOS), a prebiotic that is agglutinated by type-1 fimbriae, are proposed for the control of enterobacteria colonization and may be an alternative to Salmonella control in pigs. The aim of this study was to evaluate the capability of porcine Salmonella strains to adhere to MOS in vitro. A total of 108 strains of Salmonella sp. isolated from carrier pigs were evaluated for the amplification of fimA and fimH genes, agglutination of MOS and hemagglutination. In all tested strains, amplicons of expected size were detected for both fimA and fimH gene. In the hemagglutination assays, 31 (28.7%) strains presented mannose–sensitive agglutination of erythrocytes, indicating that the strains were expressing type-1 fimbriae. Considering only strains expressing the type-1 fimbriae, 23 (74.2%) presented a strong agglutination of MOS, 3 (9.6%) a weak reaction and 5 (16.2%) none. The results indicate that Salmonella enterica strains expressing type-1 fimbriae can agglutinate effectively in vitro to MOS. PMID:24031388

  3. 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.

  4. Detection of staphylococcal exfoliative toxin by slide latex agglutination.

    PubMed Central

    Murono, K; Fujita, K; Yoshioka, H

    1988-01-01

    A simple and rapid method in which slide latex agglutination was used was developed to detect the exfoliative toxin (ET) elaborated by clinical isolates. ET types A and B (ET-A and ET-B) were purified by plate gel isoelectrofocusing, and anti-ET sera were obtained by immunizing rabbits. A specific immunoglobulin G antitoxin was then prepared from the immunized rabbit sera by fast protein liquid chromatography, and latex particles were coated with the antitoxin. Of 74 staphylococcal strains isolated from patients with staphylococcal scalded skin syndrome, 61 strains were found to produce ET by the newborn mouse bioassay. All 61 strains were shown to be positive for ET-A and ET-B production by the slide latex agglutination method. The lowest concentration of ETs detected by the latex agglutination method was 0.5 microgram/ml, which was much lower than that detected by the double immunodiffusion method, with a sensitivity of 50 micrograms/ml. It is crucial to prove ET production by clinical isolates for the diagnosis and surveillance of staphylococcal scalded skin syndrome. The latex agglutination method is a sensitive, simple, and rapid test which can be used as an alternative to the newborn mouse bioassay. Images PMID:3343322

  5. Erythrocyte rheology.

    PubMed Central

    Stuart, J

    1985-01-01

    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. Images PMID:3900147

  6. 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.

  7. Trypsinized Human Group O Erythrocytes as an Alternative Hemagglutinating Agent for Japanese Encephalitis Virus

    PubMed Central

    Shortridge, K. F.; Hu, L. Y.

    1974-01-01

    Trypsinized human group O erythrocytes were found to be a suitable alternative to gander cells in hemagglutination (HA) and hemagglutination inhibition (HAI) tests for Japanese encephalitis (JE) virus. In the HAI test, no cross-reactions against JE virus were observed with immune sera containing antibody to taxonomically related or unrelated viruses, with mouse brain antigen, or with nonantibody serum inhibitors; specific antibody rise could be detected in an immunized rabbit. Gander and trypsinized human group O cells gave comparable titers in the HAI test, but the latter were preferable since (i) they required less challenging HA antigen, being more sensitive to agglutination by JE virus, and (ii) all human and some animal sera investigated were devoid of natural agglutinins for these cells, thereby eliminating or reducing the need for prior adsorption with packed cells. PMID:4856948

  8. A rhamnose-binding lectin from sea bass (Dicentrarchus labrax) plasma agglutinates and opsonizes pathogenic bacteria.

    PubMed

    Cammarata, Matteo; Parisi, Maria Giovanna; Benenati, Gigliola; Vasta, Gerardo R; Parrinello, Nicolò

    2014-06-01

    The discovery of rhamnose-binding lectins (RBLs) in teleost fish eggs led to the identification of a novel lectin family characterized by a unique sequence motif and a structural fold, and initially proposed to modulate fertilization. Further studies of the RBL tissue localization and gene organization were also suggestive of role(s) in innate immunity. Here we describe the purification, and biochemical and functional characterization of a novel RBL (DlRBL) from sea bass (Dicentrarchus labrax) serum. The purified DlRBL had electrophoretic mobilities corresponding to 24 kDa and 100 kDa under reducing and non-reducing conditions, respectively, suggesting that in plasma the DlRBL is present as a physiological homotetramer. DlRBL subunit transcripts revealed an open reading frame encoding 212 amino acid residues that included two tandemly-arrayed carbohydrate-recognition domains, and an 18-residue signal sequence at the N-terminus. The deduced size of 24.1 kDa for the mature protein was in good agreement with the subunit size of the isolated lectin. Binding activity of DlRBL for rabbit erythrocytes could be inhibited in the presence of rhamnose or galactose, did not require calcium, and was optimal at around 20°C and within the pH 6.5-8.0 range. DlRBL agglutinated Gram positive and Gram negative bacteria, and exposure of formalin-killed Escherichia coli to DlRBL enhanced their phagocytosis by D. labrax peritoneal macrophages relative to the unexposed controls. Taken together, the results suggest that plasma DlRBL may play a role in immune recognition of microbial pathogens and facilitate their clearance by phagocytosis.

  9. Light-scattering analysis of ultrasonic wave's influence on the RBC agglutination in vitro

    NASA Astrophysics Data System (ADS)

    Doubrovski, Valeri A.; Dvoretski, Costanten N.

    1999-04-01

    Elastic light scattering is one of the most often used optical methods to analyze the cells agglutination reaction - the base of a great number of medical diagnostic test and biomedical investigations. The increase of the resolution of methods and apparatus towards the induced cells aggregation - the foundation of the reaction of agglutination, is quite an actual problem. The solution of this problem increases the reliability of the diagnostic test and gives an opportunity to achieve the diagnostic information in the cases when the traditional approaches do not lead to the diagnostic results. The attempt to increase the resolution of the immune reaction analyzer by means of ultrasonic waves action on the reagent mixture in vitro is taken in this paper. The RBC agglutination reaction which is usually used for the blood group type examination is chosen as an example of an object of the investigation. Different laser optical trains of the devices based on the turbidimetric and nephelometric methods and their combination are analyzed here. The influence of the ultrasonic wave time interval action and of the features of the sample preparation procedure on the resolution towards the agglutination process was investigated in this work. It is shown that the ultrasonic wave action on the reagent mixture leads to a large gain in the resolution of the device towards the RBC agglutination process. The experiments showed that the resolution of the device was enough to register the agglutination process even for the erythrocytes with weak agglutination ability when the reaction was invisible without ultrasonic action. It occurred that the diagnostic test time was more than by an order shortened due to the ultrasonic wave action. The optimal ultrasonic time interval action, the sample preparation technology and experimental technique were defined. The principle of the ultrasonic wave action on the cells agglutination process suggested here can be spread out on the immune

  10. Helicobacter pylori TlyA agglutinates liposomes and induces fusion and permeabilization of the liposome membranes.

    PubMed

    Lata, Kusum; Chattopadhyay, Kausik

    2014-06-10

    Helicobacter pylori TlyA is a pore-forming hemolysin with potent cytotoxic activity. To explore the potential membrane-damaging activity of H. pylori TlyA, we have studied its interaction with the synthetic liposome vesicles. In our study, H. pylori TlyA shows a prominent ability to associate with the liposome vesicles without displaying an obligatory requirement for any protein receptor on the liposome membranes. Interaction of TlyA triggers agglutination of the liposome vesicles. Such agglutinating activity of TlyA could also be observed with erythrocytes before the induction of its pore-forming hemolytic activity. In addition to its agglutinating activity against liposomes, TlyA also induces fusion and disruption of the liposome membranes. Altogether, our study highlights novel membrane-damaging properties of H. pylori TlyA that have not been documented previously with any other TlyA family protein.

  11. The interaction of antigen and antibody in agglutination

    PubMed Central

    Elek, S. D.; Smith, B. V. Kingsley; Highman, Wilma

    1964-01-01

    Flagellar fragments are thin cylinders that are particularly suitable for the study of agglutination by electron microscopy. Their shape leads to a characteristic pattern of agglutination and thus the early stages can be studied. Measurement of interflagellar distances under conditions of negative staining, suggest that the minimum length of the rabbit antibody molecules is about 180 Å. The molecules carry the specific sites at the ends of the long axis, and become attached radially to the surface of the flagella, resembling the bristles of a bottle-brush. To explain this orientation it is postulated that the antibody is inserted into the surface of the flagellum in such a manner that surrounding molecules give it a fixed direction. Geometrically this hypothesis corresponds to an insertion into pits. Pepsin-treated (5S) rabbit antibody behaves in a like manner, but the molecule appears to be shorter. No information could be obtained about the thickness and actual shape of antibody molecules by the techniques employed. ImagesFIG. 1FIG. 2FIG. 3FIG. 4FIG. 5FIG. 6FIG. 9FIG. 11FIG. 13FIG. 14FIG. 15 PMID:14210767

  12. 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

  13. Agglutination of Helicobacter pylori coccoids by lectins

    PubMed Central

    Khin, Mar Mar; Hua, Jie Song; Ng, Han Cong; Wadström, Torkel; Ho, Bow

    2000-01-01

    AIM: To study the agglutination pattern of Helicobacter pylori coccoid and spiral forms. METHODS: Assays of agglutination and agglutination inhibition were applied using fifteen commercial lectins. RESULTS: Strong agglutination was observed with mannose-specific Concanavalin A (Con A), fucose-specific Tetragonolobus purpureas (Lotus A) and N-acetyl glucosamine-specific Triticum vulgaris (WGA) lectins. Mannose and fucose specific lectins were reactive with all strains of H. pylori coccoids as compared to the spirals. Specific carbohydrates, glycoproteins and mucin were shown to inhibit H. pylori lectin-agglutination reactions. Pre-treatment of the bacterial cells with formalin and sulphuric acid did not alter the agglutination patterns with lectins. However, sodium periodate treatment of bacterial cells were shown to inhibit agglutination reaction with Con A, Lotus A and WGA lectins. On the contrary, enzymatic treatment of coccoids and spirals did not show marked inhibition of H. pylori lectin agglutination. Interes tingly, heating of H. pylori cells at 60 °C for 1 h was shown to augment the agglutination with all of the lectins tested. CONCLUSION: The considerable differences in lectin agglutination patterns seen among the two differentiated forms of H. pylori might be attributable to the structural changes during the events of morphological transformation, resulting in exposing or masking some of the sugar residues on the cell surface. Possibility of various sugar residues on the cell wall of the coccoids may allow them to bind to different carbohydrate receptors on gastric mucus and epithelial cells. The coccoids with adherence characteristics like the spirals could aid in the pathogenic process of Helicobacter infection. This may probably lead to different clinical outcome of H. pylori associated gastroduodenal disease. PMID:11819557

  14. Development of a slide agglutination assay for detection of blastomycosis.

    PubMed

    Hatch, Wayne O; Scalarone, Gene M

    2013-11-01

    Blastomycosis, caused by the thermally dimorphic fungus Blastomyces dermatitides, which is endemic to eastern regions of the USA, is commonly misdiagnosed as a viral or bacterial infection and therefore treated improperly. Over the years, many immunodiagnostic assays to aid in the diagnosis of blastomycosis have been developed; however, a reliable assay for use in local clinics still remains elusive. Procedures for a slide agglutination assay for detection of antibody in serum from rabbits immunized with B. dermatitidis were evaluated with antigenic preparations from B. dermatitidis adsorbed to polystyrene microparticles. Yeast-phase lysates from five isolates of B. dermatitides: namely ER-593 (Eagle River, WI, USA), ER-598 (Eagle River, WI, USA), 48938 (India), B5896 (Mt. Iron, MN, USA), and T-58 (TN, USA) were evaluated for their sensitivity and specificity. Sensitivities of the lysates ranged from 29% to 83% whereas specificities ranged from 13% to 100%. Lysate ER-593 provided the most promising results with a sensitivity of 82% and specificity of 100%. This study provides suggests that a simple rapid slide agglutination assay for detecting blastomycosis may be used for screening patients with suspected B. dermatitidis infection.

  15. R and G color component competition of RGB image decomposition as a criterion to register RBC agglutinates for blood group typing.

    PubMed

    Doubrovski, Valeri A; Ganilova, Yuliya A; Zabenkov, Igor V

    2014-03-01

    A new approach of the criterion assignment for registration of erythrocyte agglutinates to instrumentally determine blood group type is suggested. The criterion is based on comparison of R and G components of RGB decomposition of microscopy digital image taken for the blood-serum mixture sample. For the chosen experimental conditions, the minimal size (area) of RBC agglutinate to be registered by the criterion suggested is estimated theoretically. The proposed method was tested experimentally on the example of monitoring agglutinates in flow. The encouraging experimental results were obtained for improvement of the resolving power of the method; the optimal experimental conditions were revealed for maximum resolution. Though the suggested method was realized for dynamic (flow) blood group determination, it could also be applied for diagnostics in a stationary environment. This approach increases the reliability of RBC agglutinates registration and, hence, blood group typing. The results may be used to develop the apparatus for automated determination of human blood group.

  16. Rapid detection of methicillin-resistant Staphylococcus aureus strains not identified by slide agglutination tests.

    PubMed Central

    Kuusela, P; Hildén, P; Savolainen, K; Vuento, M; Lyytikäinen, O; Vuopio-Varkila, J

    1994-01-01

    Seventy-nine methicillin-resistant Staphylococcus aureus (MRSA) strains, isolated during 1980 to 1990, were classified as MRSA Aggl- (14 strains) and MRSA Aggl+ (65 strains) strains on the basis of test results in slide agglutination assays designed to detect fibrinogen-binding protein (clumping factor) and protein A on the staphylococcal surface. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis revealed that lysostaphin digests of MRSA Aggl- strains contained a high-molecular-weight protein which was not detected in digests of MRSA Aggl+ strains. Immunization of rabbits with an MRSA Aggl- strain produced an antiserum which agglutinated all MRSA Aggl- strains and also 64 of 65 MRSA Aggl+ strains. Only 1 of 68 coagulase-negative staphylococci showed agglutination in this assay. The anti-MRSA Aggl- antiserum reacted mainly with a 230-kDa staphylococcal surface protein but also with a 175-kDa protein, probably formed by proteolysis of the former and a few slightly smaller proteins. These could not be immunologically detected in lysostaphin digests of MRSA Aggl+ strains. Purified antibodies reacting with the 230-kDa protein agglutinated all MRSA Aggl- strains, indicating that the protein is located on the surfaces of staphylococci. The results suggest a tentative role for the 230-kDa protein or its fragments as a novel target to develop more efficient rapid identification methods for S. aureus, including MRSA. Images PMID:8126170

  17. Simulation of Mechanical Behavior of Agglutinates

    NASA Technical Reports Server (NTRS)

    Nakagawa, Masami; Moon, Tae-Hyun

    2005-01-01

    Due to lack of "real" lunar soil or even lunar simulant, it is difficult to characterize the interaction between lunar soil (or simulant) with different surfaces that are involved in excavation and processing machinery. One unique feature possessed by lunar soil is the agglutinates produced by repeated high-speed micrometeoroid impacts and subsequent pulverization[l and 2]. The large particles are impacted by micrometeoroids [Fig.l] and pulverized to produce finer particles. This process continues until there are no more "large" particles left on the surface of the moon. Due to high impact speed, the impact melting process fuses fines to make agglutinates such as shown in Fig. 2. We will present a series of simulation results and movies will be shown to indicate brittle behavior of each individual agglutinate and also similar compressibility charts shown by Carrier et al. [3]. Fig. 3 shows our preliminary result of the simulated oedometer tests.

  18. Stable suspension for Vi-agglutination tests

    PubMed Central

    Ando, Koji; Shimojo, Hiroto

    1953-01-01

    Two methods of preparing a stable suspension for Vi-agglutination tests are discussed. Both maintain Vi-agglutinability and O-inagglutinability after storage at 37°C for 6 months, and the second also maintains the Vi-capsule-staining property. The first method involves the addition of 0.5% CaCl2 to a heavy saline Vi-suspension, while in the second a similar suspension is treated with an 0.2% solution of chrome alum. PMID:20603972

  19. Are the Major Agglutinative Languages Genetically Related?

    ERIC Educational Resources Information Center

    Hakola, H. P. A.

    1989-01-01

    Examination of accidental CVC and CV correspondences among languages representing 5 large families of agglutinative languages found that comparison pairs had much more similarity between basic 100-word vocabularies than would have been possible by mere chance, supporting the hypothesis that those 5 language families were mutually related.…

  20. Process to create simulated lunar agglutinate particles

    NASA Technical Reports Server (NTRS)

    Gustafson, Robert J. (Inventor); Gustafson, Marty A. (Inventor); White, Brant C. (Inventor)

    2011-01-01

    A method of creating simulated agglutinate particles by applying a heat source sufficient to partially melt a raw material is provided. The raw material is preferably any lunar soil simulant, crushed mineral, mixture of crushed minerals, or similar material, and the heat source creates localized heating of the raw material.

  1. A technique for detection of agglutinating activity of antilymphocytic serum using formalinized and trypan-blue-stained lymphocytes

    PubMed Central

    Janković, B. D.; Isaković, Katarina; Petrović, Spomenka

    1970-01-01

    This paper describes technical details of a micro-reaction for the in vitro detection of agglutinating potency of antilymphocytic serum produced in rabbits with chicken, rat and dog thymus cells. The titration of antilymphocytic serum includes the use of microtitre plastic plates and lymphocytes treated with formalin and stained with trypan blue stain. Formalinized and stained lymphocytes represent a stable antigen of long durability, and the application of those cells in leucoagglutination increases the accuracy and sensitivity of the reaction. PMID:5477930

  2. [Identification of mosquitoes' human food source by using the co-agglutination technique].

    PubMed

    Castex, M; Fachado, A; Fonte, L

    1997-01-01

    The utilization of a coagglutination technique for the identification of a human source for feeding mosquitoes is described. The dilution of ingested blood samples in filter paper was performed in 2 mL of a sodium chloride solution at 0.85%. It was used a suspension of sensibilized Staphylococcus aureus with rabbit's serum, human plasmatic anti-proteins, and human anti-IgG rabbit's serum discriminated well between human and non human blood. No agglutination was observed with the negative control. This technique proved to be sensitive to identify 100% of the human blood samples taken to the paper 24 hours after the mosquitoes completed their feeding at a temperature of 26 to 28 degrees C. Among mosquitoes fed and collected in the fields the test had a satisfactory result. Therefore, it may be used in routine work in the fields. The results showed the sensitivity and specificity of this method for identifying human blood ingested by mosquitoes.

  3. The chemistry of some individual lunar soil agglutinates

    NASA Technical Reports Server (NTRS)

    Gibbons, R. V.; Hoerz, F.; Schaal, R. B.

    1976-01-01

    The inquiry is centered on the composition of agglutinate glasses examined via microprobe techniques. The glass chemistry of the agglutinates is brought into relation with compositions of constituent detritus and bulk compositions of the parent soils, with recent reported results taken into cognizance. Electron microprobe analysis data were examined for possible chemical fractionation resulting from meteoritic impacts and formation of agglutinates in the lunar regolith; individual agglutinates from lunar soils 78222, 71061, and 60009 were probed. Differences between impact glasses and corresponding bulk soils were scrutinized. Agglutinate glass analyses tend to cluster near the bulk soil compositions. A slight enrichment in mafic elements in grand averages of the agglutinate clusters relative to the bulk soils was found. Evidence of total impact melts and minor partial shock melts is examined.

  4. Mass-based readout for agglutination assays

    NASA Astrophysics Data System (ADS)

    Chunara, Rumi; Godin, Michel; Knudsen, Scott M.; Manalis, Scott R.

    2007-11-01

    We present a mass-based readout for agglutination assays. The suspended microchannel resonator (SMR) is used to classify monomers and dimers that are formed during early stage aggregation, and to relate the total count to the analyte concentration. Using a model system of streptavidin functionalized microspheres and biotinylated antibody as the analyte, we obtain a dose-response curve over a concentration range of 0.63-630nM and show that the results are comparable to what has been previously achieved by image analysis and conventional flow cytometry.

  5. Penicillamine prevents ram sperm agglutination in media that support capacitation.

    PubMed

    Leahy, T; Rickard, J P; Aitken, R J; de Graaf, S P

    2016-02-01

    Ram spermatozoa are difficult to capacitate in vitro. Here we describe a further complication, the unreported phenomenon of head-to-head agglutination of ram spermatozoa following dilution in the capacitation medium Tyrodes plus albumin, lactate and pyruvate (TALP). Sperm agglutination is immediate, specific and persistent and is not associated with a loss of motility. Agglutination impedes in vitro sperm handling and analysis. So the objectives of this study were to investigate the cause of sperm agglutination and potential agents which may reduce agglutination. The percentage of non-agglutinated, motile spermatozoa increased when bicarbonate was omitted from complete TALP suggesting that bicarbonate ions stimulate the agglutination process. d-penicillamine (PEN), a nucleophilic thiol, was highly effective at reducing agglutination. The inclusion of 250 μM PEN in TALP reduced the incidence of motile, agglutinated spermatozoa from 76.7 ± 2.7% to 2.8 ± 1.4%. It was then assessed if PEN (1 mM) could be included in existing ram sperm capacitation protocols (TALP +1 mM dibutyryl cAMP, caffeine and theophylline) to produce spermatozoa that were simultaneously capacitated and non-agglutinated. This protocol resulted in a sperm population which displayed high levels of tyrosine phosphorylated proteins and lipid disordered membranes (merocyanine-540) while remaining motile, viable, acrosome-intact and non-agglutinated. In summary, PEN (1 mM) can be included in ram sperm capacitation protocols to reduce sperm agglutination and allow for the in vitro assessment of ram sperm capacitation.

  6. Direct agglutination test for serologic diagnosis of Neospora caninum infection.

    PubMed

    Romand, S; Thulliez, P; Dubey, J P

    1998-01-01

    A direct agglutination test was evaluated for the detection and quantitation of IgG antibodies to Neospora caninum in both experimental and natural infections in various animal species. As compared with results obtained by the indirect fluorescent antibody test, the direct agglutination test appeared reliable for the serologic diagnosis of neosporosis in a variety of animal species. The direct agglutination test should provide easily available and inexpensive tools for serologic testing for antibodies to N. caninum in many host species.

  7. Nitrogen isotopic signatures in agglutinates from breccia 79035

    NASA Technical Reports Server (NTRS)

    Kerridge, John F.; Kim, Yoosook; Kim, Jin S.; Marti, Kurt

    1993-01-01

    Agglutinates in the size range 125-175 microns from regolith breccia 79035 are substantially depleted in N compared with bulk 79035. Isotopically, agglutinate N closely resembles that found previously in ilmenite separates. The minimum (delta)N-15 value found during stepwise pyrolysis of agglutinates is significantly heavier than that observed for bulk 79035. The major host phase for trapped N in 79035, and the host phase of the lightest isotopic component(s), remain unidentified.

  8. Rabbit medicine.

    PubMed

    Jordan, Dinah G

    2007-01-01

    When filling prescriptions for a rabbit, it is important to know whether the rabbit is a pet or is being raised as a source of food for human consumption. Several drugs widely used for pet rabbits are prohibited from exralabel use in animals raised for food production. The list of banned drugs should always be perused prior to filling a prescription for a rabbit being raised for food production. Since no veterinary-approved products exist for rabbits and most medications must be compounded, pharmacists are likely to encounter prescriptions for rabbits in their practice. A basic understanding of rabbit anatomy, physiolgy and common diseases will assist pharmacists in distinguishing between safe and dangerous drugs for administration to rabbits.

  9. 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.

  10. Disposable rabbit

    DOEpatents

    Lewis, Leroy C.; Trammell, David R.

    1986-01-01

    A disposable rabbit for transferring radioactive samples in a pneumatic transfer system comprises aerated plastic shaped in such a manner as to hold a radioactive sample and aerated such that dissolution of the rabbit in a solvent followed by evaporation of the solid yields solid waste material having a volume significantly smaller than the original volume of the rabbit.

  11. Disposal rabbit

    DOEpatents

    Lewis, L.C.; Trammell, D.R.

    1983-10-12

    A disposable rabbit for transferring radioactive samples in a pneumatic transfer system comprises aerated plastic shaped in such a manner as to hold a radioactive sample and aerated such that dissolution of the rabbit in a solvent followed by evaporation of the solid yields solid waste material having a volume significantly smaller than the original volume of the rabbit.

  12. Blood stained cerebrospinal fluid responsible for false positive reactions of latex particle agglutination tests.

    PubMed Central

    Camargos, P A; Almeida, M S; Filho, G L; Batista, K W; Carvalho, A G; Pereira, C L

    1994-01-01

    The accuracy of the latex particle agglutination test (LPAT) was assessed in blood stained cerebrospinal fluid (CSF) specimens from 166 paediatric patients, aged from three months to 13 years. A commercial LPAT kit was used to detect Haemophilus influenzae type b, Streptococcus pneumoniae, and Neisseria meningitidis A, B, and C soluble antigens. Culture of CSF specimens was used as the standard and all laboratory procedures were performed blind. The mean CSF erythrocyte count was 66,406 cells/mm3 in the cases and 11,560 cells/mm3 in the controls. The sensitivity and the specificity of LPAT were 83.8 and 94.0%, respectively, suggesting that LPAT is a useful diagnostic tool even in blood stained CSF specimens. PMID:7876387

  13. Systems, devices, and methods for agglutination assays using sedimentation

    DOEpatents

    Schaff, Ulrich Y.; Sommer, Gregory J.; Singh, Anup K.

    2016-01-26

    Embodiments of the present invention include methods for conducting agglutination assays using sedimentation. Aggregates may be exposed to sedimentation forces and travel through a density medium to a detection area. Microfluidic devices, such as microfluidic disks, are described for conducting the agglutination assays, as are systems for conducting the assays.

  14. Latex agglutination assays for detection and of non-O157 Shiga toxin-producing E. coli serogroups O26, O45, O103, O111, O121 and O145

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Latex agglutination assays were developed for the top six non-O157 Shiga toxin-producing Escherichia coli (STEC) serogroups utilizing polyclonal antibodies. Rabbit antisera were affinity purified through Protein A/G columns and the isolated immunoglobulins (IgG) were covalently immobilized onto pol...

  15. 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.

  16. Binding Specificity of Piliated Strains of Escherichia coli and Salmonella typhimurium to Epithelial Cells, Saccharomyces cerevisiae Cells, and Erythrocytes

    PubMed Central

    Korhonen, Timo K.; Leffler, Hakon; Edén, Catharina Svanborg

    1981-01-01

    The binding to mammalian cells of piliated enteric bacteria and the inhibition of the binding by antibodies to purified pili were studied. The target cells were epithelial cells from human bucca and human and rat urinary tracts, erythrocytes from various species, and Saccharomyces cerevisiae cells. The strains were selected to represent the two main agglutination patterns of enteric bacteria: mannose-resistant agglutination of human and other erythrocytes and mannose-sensitive agglutination of guinea pig and other erythrocytes. Escherichia coli 3669 caused only mannose-resistant agglutination, E. coli 6013 caused only mannose-sensitive agglutination, and E. coli 3048 caused both types of agglutination simultaneously. Salmonella typhimurium SH6749 exhibited only mannose-sensitive hemagglutination and was included to allow comparison of its pili with those of E. coli strains. The range of epithelial cells to which the bacteria adhered was related to their agglutination patterns. All four strains attached to human buccal cells. Only E. coli strains 3669 and 3048, which caused mannose-resistant agglutination, adhered to human urinary tract epithelial cells, and only those strains that caused mannose-sensitive agglutination adhered to rat urinary tract epithelial cells. The binding of S. typhimurium SH6749, but not of the other strains with mannose-sensitive agglutination, was significantly inhibited by d-mannose. Globotetraosylceramide, a glycolipid present in the human urinary tract epithelium, inhibited attachment to human uroepithelial cells of the two strains with mannose-resistant hemagglutination. As tested by the enzyme-linked immunosorbent assay, cross-reactions between type 1 pili of the E. coli strains were strong, but those between S. typhimurium and E. coli mannose-sensitive pili were weak. The two pili that induced mannose-resistant hemagglutination on E. coli did not cross-react. Significant inhibition of adhesion of all four strains was obtained with the

  17. Moraxella bovis hemagglutinins: effect of carbohydrates, heating and erythrocytes.

    PubMed Central

    Gil-Turnes, C; Ribeiro, G A

    1985-01-01

    Several properties of the adhesins of eight isolates of Moraxella bovis recovered from cattle suffering from infectious keratoconjunctivitis, were studied. Adhesions were detected through autoagglutination in saline and hemagglutination. Autoagglutinating strains agglutinated red blood cells of the chicken, rabbit, sheep and swine, but not those of the guinea pig. The adhesins were not inhibited by D-mannose or D-galactose and resisted heating at 100 degrees C for 15 minutes. Magnesium chloride at a final concentration of 10% inhibited autoagglutination and hemagglutination. The value of the hemagglutination test for monitoring synthesis of fimbriae by M. bovis, is discussed. PMID:3986674

  18. Prevention of nonspecific reactions on reversed passive latex agglutination assay (RPLA) for detecting low amounts of staphylococcal enterotoxins.

    PubMed

    Pereira, M L; Heneine, L G; Santos, E J; Carmo, L S; Pereira, J L; Bergdoll, M S

    1997-01-01

    The SET-RPLA, from Denka Seiken Co. Ltd., Tokio, a commercial reversed passive latex agglutination test kit, has been recommended to establish the enterotoxicity capacity of some staphylococcal strains, implicated in food poisoning outbreaks that produce low levels of enterotoxins (SE). Despite the RPLA specificity, the occurrence of nonspecific reactions when testing low-SE-producing is common. In order to control these nonspecific reactions the addition of purified normal rabbit IgG purified was applied on approximately 350 staphylococcal isolates from human milk and anatomic sites of healthy dental student carriers. The results indicated that addition of 5% (v/v) of purified normal rabbit IgG (0.74 mg/mL) to the culture supernatant fluid is a simple and reliable tool for the controlling of nonspecific reactions in the RPLA assay.

  19. [COMPARATIVE STUDY OF MECHANICAL STRESS EFFECT ON HUMAN AND ANIMAL ERYTHROCYTES].

    PubMed

    Shpakova, N M; Orlova, N V; Nipot, E E; Aleksandrova, D I

    2015-01-01

    Sensitivity of human and animal (bovine, rat, rabbit, equine) erythrocytes to the effect of mechanical stress has been studied. Mechanical stress effect was demonstrated to result in a time-dependent (5-60 min) release of potassium cations out of mammalian erythrocytes and a partial hemolytic cell damage. Herewith the release levels of potassium ions and hemolysis did not coincide for erythrocytes of all the mammals except rabbit ones. The most sensitive to mechanical stress (60 min) by the parameters of hemolytic damage and potassium ion release were rat (32%) and bovine (66%) erythrocytes respectively, the lowest sensitive by both parameters were rabbit ones (about 20%). Implemented correlation analysis has demonstrated a statistically significant negative relation between the values of mechanical hemolysis of mammalian erythrocytes and surface-volumetric ratio of cells (rs = -0.900, P = 0.037). A feasible relationship between the content of phosphatidylethanolamine in mammalian erythrocyte membranes and the level of potassium cation loss under mechanical stress effect is under discussion.

  20. 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.

  1. Seroprevalence of Toxoplasma gondii infection in domestic rabbits in Durango State, Mexico.

    PubMed

    Alvarado-Esquivel, Cosme; Alvarado-Esquivel, Domingo; Villena, I; Dubey, J P

    2013-09-01

    There is a lack of information concerning the seroprevalence of Toxoplasma gondii infection in rabbits in northern Mexico. Through a cross sectional study, antibodies to T. gondii were determined in 429 domestic rabbits in Durango State, Mexico using the modified agglutination test. Rabbits were raised in 29 properties in 6 municipalities. Overall, antibodies to T. gondii were found in 70 (16.3%) of 429 rabbits, with titers of 1:25 in 42, 1:50 in 19, 1:100 in 5, 1:200 in 3, and 1:800 in 1. Seropositive rabbits were found in 21 (72.4%) of 29 properties, including 16 of 21 homes, 4 of 5 farms, and 1 of 3 pet shops. This is the first study of T. gondii infection in rabbits in Durango, Mexico. Results indicate that infected rabbits are a potential source of T. gondii infection in humans in Durango State.

  2. Glycolipid receptors for uropathogenic Escherichia coli on human erythrocytes and uroepithelial cells.

    PubMed Central

    Leffler, H; Svanborg-Edén, C

    1981-01-01

    A specific family of glycolipids, the globoseries, was shown to act as receptors on human uroepithelial cells and erythrocytes for the majority of uropathogenic Escherichia coli strains attaching to or hemagglutinating those cells. This was demonstrated in three different ways: (i) correlation between the natural presence of glycolipid in the target cell (erythrocytes of different species) and binding of bacteria; (ii) inhibition of attachment to human uroepithelial cells by preincubation of bacteria and glycolipid; and (iii) induction of binding to unreactive cells by coating of these cells with glycolipid. Strains reacting with the receptor agglutinated guinea pig erythrocytes in a mannose-resistant way after, but not before, coating of the cells with globotetraosylceramide. Unrelated glycolipids were not recognized. The reaction was made independent of simultaneous occurrence of mannose-sensitive adhesions on the strains by addition of D-mannose. The receptor-coated cells were used as a tool to screen for prevalence of receptor recognition in a collection of 453 E. coli strains isolated from patients with urinary tract infection or from the stools of healthy children. Of 150 strains attaching to human uroepithelial cells and agglutinating human erythrocytes, 121 bound to globotetraosylceramide (81%). Globoside recognition was especially frequent among pyelonephritis strains (74/81). The glycolipid composition of the urogenital epithelium and kidney tissue and the ability of uropathogenic E. coli to bind to these glycolipids may be a determinant in host-parasite interaction leading to urinary tract infection. PMID:7037645

  3. 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.

  4. Tritrichomonas foetus: a scanning electron microscopy study of erythrocyte adhesion associated with hemolytic activity.

    PubMed

    De Carli, Geraldo Attilio; Tasca, Tiana; Pires Borges, Fernanda

    2004-01-01

    The in vitro hemolytic activity of Tritrichomonas foetus was investigated. The parasite was tested against human erythrocytes of groups A, B, AB, and O, and against erythrocytes of nine adult animals of different species (the rabbit, rat, chicken, cat, dog, swine, horse, bovine, and sheep). The results showed that T. foetus strains (ATCC KV1, K, PAL, 5022, RJ, 90) did not present any hemolytic activity against any human erythrocyte group nor against rabbit, rat, chicken, cat, dog and swine erythrocytes. T. foetus strains, however, lysed horse, bovine, and sheep erythrocytes. No hemolysin released by the parasites could be identified. Hemolysis did not occur with trichomonad culture supernatants, with sonicated extracts of T. foetus, nor with killed organisms. Scanning electron microscopy (SEM) showed that human erythrocytes did not adhere to the trophozoites, in contrast horse erythrocytes adhered to the surface of the parasites and were phagocytosed for up to 90 min. The parasites are able to exert their cytopathic effects through: (a) physical contact established between the two cell surfaces, (b) toxins released from parasites into the interaction media, or (c) the association of both mechanisms. Further studies are necessary to clarify the importance of the hemolytic activity in the biology of T. foetus.

  5. Interferon and antibody titrations using haemagglutinating Togaviridae and trypsinized human erythrocytes.

    PubMed

    Sedmak, J J; Dixon, M; Schoenherr, C; Sabran, J L; Grossberg, S E

    1983-02-01

    Several Togaviridae of the alphavirus and flavivirus genera agglutinate trypsinized human group O erythrocytes (THOE) (Shortridge and Hu, 1976). Haemagglutinin titers of Semliki Forest virus (SFV) and Japanese encephalitis virus (JEV) measured with THOE were equivalent to, if not higher than, those obtained with Embden gander erythrocytes, even with unextracted haemagglutinin. Results obtained with THOE in JEV haemagglutination-inhibition tests on sera taken from a previously infected individual over a 20-yr period were similar to those measured during the initial JEV infection. The inhibition of SFV haemagglutinin production as measured with THOE was a very sensitive bioassay for chicken interferon: interferon titers were 6- to 10-fold higher than those obtained with the vesicular stomatitis virus plaque-reduction method. The generally greater availability of human erythrocytes (including those stabilized with glutaraldehyde), the simplicity of the trypsin treatment, and the possibility of using unextracted haemagglutinin recommend this technique for use with haemagglutinating Togaviridae.

  6. 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.

  7. Improving agglutination tests by working in microfluidic channels.

    PubMed

    Degré, G; Brunet, E; Dodge, A; Tabeling, P

    2005-06-01

    Latex agglutination tests are used for the diagnosis of diseases in man and animals. They are generally simple, cheap, and do not require sophisticated equipment, nor highly specialized skills. In this Technical Note, we put latex agglutination tests in a microfluidic format. The experiment is performed in PDMS (polydimethylsiloxane) microchannels, using streptavidin-coated superparamagnetic beads and a magnetic field. The target molecule is biotinylated protein A. By taking full advantage of the microfluidic conditions (scaling down of the detection volume and controlled action of the shear flow), we achieved an analytical sensitivity of 10 fmol l(-1)(several hundreds of fg ml(-1)) and a fast response (a few minutes) ; the test is also quantitative. Performances of agglutination tests can thus be improved by orders of magnitude by adapting them to a microfluidic format; this comes in addition to the usual advantages offered by this technology (integration, high throughput etc.).

  8. Quantification of IgG on erythrocytes of patients and normals by a radio-ligand-binding assay.

    PubMed

    Giles, C M; Davies, K A; Loizou, S; Moulds, J J; Walport, M J

    1991-12-01

    A monoclonal IgG anti-human IgG, 1B12, was used in a radio-ligand-binding assay to quantify IgG on erythrocytes of patients and normals. The assay detected a range of 10-700 IgG molecules. Good correlation was achieved between the number of molecules and the strength of agglutination in antiglobulin tests performed in capillary tubes. The assay was capable of detecting subagglutinating immune bound IgG on erythrocytes from patients with systemic lupus erythematosus (SLE).

  9. Erythrocyte binding preference of 16 subtypes of low pathogenic avian influenza and 2009 pandemic influenza A (H1N1) viruses.

    PubMed

    Wiriyarat, Witthawat; Lerdsamran, Hatairat; Pooruk, Phisanu; Webster, Robert G; Louisirirotchanakul, Suda; Ratanakorn, Parntep; Chaichoune, Kridsada; Nateerom, Kannika; Puthavathana, Pilaipan

    2010-12-15

    All 16 subtypes of avian influenza viruses of low pathogenicity (LPAIV) as well as their hemagglutinin (H) antigens, and four 2009 pandemic influenza A (H1N1) virus isolates were assayed for hemagglutinating activity against 5 erythrocyte species: goose, guinea pig, human group O, chicken and horse. Of all viruses and antigens assayed, the highest hemagglutination (HA) titers were obtained with goose and guinea pig erythrocytes. Hemagglutinating activity of replicating LPAIV and LPAIV antigens decreased, in order, with chicken and human group O; meanwhile, horse erythrocytes yielded lowest or no HA titer. Moreover, the 2009 pandemic viruses did not agglutinate both horse and chicken erythrocytes. Our study concluded that goose and guinea pig erythrocytes are the best in HA assay for all subtypes of influenza viruses.

  10. Evolution of Shock Melt Compositions in Lunar Agglutinates

    NASA Technical Reports Server (NTRS)

    Vance, A. M.; Christoffersen, R.; Keller, L. P.

    2015-01-01

    Lunar agglutinates are aggregates of regolith grains fused together in a glassy matrix of shock melt produced during smaller-scale (mostly micrometeorite) impacts. Agglutinate formation is a key space weathering process under which the optically-active component of nanophase metallic Fe (npFe(sup 0)) is added to the lunar regolith. Here we have used energy-dispersive X-ray (EDX) compositional spectrum imaging in the SEM to quantify the chemical homogeneity of agglutinitic glass, correlate its homogeneity to its parent soil maturity, and identify the principle chemical components contributing to the shock melt compositional variations.

  11. 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.

  12. 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.

  13. Failure to Influence the Rejection Time of Homologous Skin Grafts in the Rabbit by Prior Injection of Donor Blood*

    PubMed Central

    Piomelli, S.; Brooke, M. S.

    1961-01-01

    Intravenous injection of rabbits with homologous whole blood or erythrocytes did not influence the rejection time of subsequent grafts from the blood donors. In some animals warm agglutinins developed, and donor erythrocytes labelled with chromium had an immune disappearance slope. PMID:14486825

  14. Rapid detection of human group C rotaviruses by reverse passive hemagglutination and latex agglutination tests using monoclonal antibodies.

    PubMed Central

    Kuzuya, M; Fujii, R; Hamano, M; Nagabayashi, T; Tsunemitsu, H; Yamada, M; Nii, S; Mori, T

    1993-01-01

    Reverse passive hemagglutination (RPHA) tests and a latex agglutination test with monoclonal antibodies (MAbs) were developed for the rapid detection of noncultivatable human group C rotaviruses. For RPHA tests, two MAbs, MAb 5A12 recognizing the outer capsid and MAb 13A3 recognizing the inner capsid, were separately used for the coating of sheep erythrocytes (SRBCs). Forty-six fecal samples were examined to confirm the practicality of the tests. As a result, there was concordance between the RPHA test with SRBCs coated with MAb 5A12 and polyacrylamide gel electrophoresis of viral RNA (RNA-PAGE) in 44 (95.6%) of 46 samples, while the diagnoses by the RPHA test with SRBCs coated with MAb 13A3 were in complete agreement with those by RNA-PAGE. Furthermore, a latex agglutination test with MAb 13A3 was also developed, and this test was fast enough and sensitive enough to successfully detect the viruses from most fecal samples within 2 min. The present procedures would be useful for the diagnosis of human group C rotavirus infections in clinical laboratories which are not well equipped. Images PMID:8388891

  15. [The pretransfusion bedside agglutination test is not a "Gold Standard"].

    PubMed

    Levy, G

    2008-11-01

    ABO-incompatible transfusions remain frequent in Europe despite the technological progresses in relation with the potential number of human errors during the control procedures of the transfusion chain. The agglutination bedside-test is only one step of this chain and the amelioration of the security will be achieved by its replacement by an electronical check.

  16. Synthesis for Lunar Simulants: Glass, Agglutinate, Plagioclase, Breccia

    NASA Technical Reports Server (NTRS)

    Weinstein, Michael; Wilson, Stephen A.; Rickman, Douglas L.; Stoeser, Douglas

    2012-01-01

    The video describes a process for making glass for lunar regolith simulants that was developed from a patented glass-producing technology. Glass composition can be matched to simulant design and specification. Production of glass, pseudo agglutinates, plagioclase, and breccias is demonstrated. The system is capable of producing hundreds of kilograms of high quality glass and simulants per day.

  17. Comparative Study of Erythrocyte Glycolytic Enzymes in Man and in Two Species of Lagomorphae

    PubMed Central

    Puget, A.; Gouarderes, Christine; Vergnes, H.

    1974-01-01

    The authors made a comparative study of red cell enzyme glycolysis in man and in two species of Lagomorphae, the pika and the rabbit. The activities of the 12 enzymes of Embden-Meyerhoff pathway and of the two dehydrogenases of pentose shunt (glucose-6-phosphate dehydrogenase and 6-phosphogluconate dehydrogenase) were determined. Phosphoglycerate kinase and pyruvate kinase showed quite similar activities in pika erythrocytes and in erythrocytes from human umbilical cord. The levels of these enzymes differed significantly in the pika and in the rabbit. No differences were noted between pyruvate kinases from the rabbit and the neonatal man. The other activities gave values either identical to those found in the adult man or intermediate between the adult and the neonatal man. In the rabbit the levels of glycolitic enzymes were generally lower than in the pika except for lactate dehydrogenase and glycose-6-phosphate dehydrogenase. Some characteristics of erythrocyte metabolism specific to the pika may account for the differences observed in this species. The influence of red cell age cannot explain the variations observed for no significant reticulocytosis was observed in the circulating blood. The percentages found in the pika and the rabbit were essentially identical at determination. PMID:4277413

  18. Immunoreactions involving platelets. III. Quantitative aspects of platelet agglutination, inhibition of clot retraction, and other reactions caused by the antibody of quinidine purpura.

    PubMed

    SHULMAN, N R

    1958-05-01

    Quantitative aspects of platelet agglutination and inhibition of clot retraction by the antibody of quinidine purpura were described. The reactions appeared to depend on formation of types of antibody-quinidine-platelet complexes which could fix complement but complement was not necessary for these reactions. Complement fixation was at least 10 times more sensitive than platelet agglutination or inhibition of clot retraction for measurement and detection of antibody activity. Although it has been considered that antibodies of drug purpura act as platelet lysins in the presence of complement and that direct lysis of platelets accounts for development of thrombocytopenia in drug purpura, the present study suggests that attachment of antibody produces a change in platelets which is manifested in vitro only by increased susceptibility to non-specific factors which can alter the stability of platelets in the absence of antibody. The attachment of antibody to platelets in vivo may only indirectly affect platelet survival. In contrast to human platelets, dog, rabbit, and guinea pig platelets, and normal or trypsin-treated human red cells did not agglutinate, fix complement, or adsorb antibody; and intact human endothelial cells did not fix complement or adsorb antibody. Rhesus monkey platelets were not agglutinated by the antibody but did adsorb antibody and fix complement although their activity in these reactions differed quantitatively from that of human platelets. Cinchonine could be substituted for quinidine in agglutination and inhibition of clot retraction reactions but quinine and cinchonidine could not. Attempts to cause passive anaphylaxis in guinea pigs with the antibody of quinidine purpura were not successful.

  19. Agglutinates as recorders of regolith evolution - Application to the Apollo 17 drill core

    SciTech Connect

    Laul, J.C.; Smith, M.R.

    1984-11-15

    Chemical data are reported for agglutinates from 26 depth intervals of the Apollo 17 deep drill core, and the compositions of the agglutinates are compared with those of the soils in which they occur. The agglutinate sequence suggests a scenario in which several closely-spaced depositional events were involved in the formation of the drill core, rather than a continuous accumulation process.

  20. Anemia and mechanism of erythrocyte destruction in ducks with acute Leucocytozoon infections

    USGS Publications Warehouse

    Kocan, R.M.

    1968-01-01

    In the anemia which accompanies infection by Leucocytozoon simondi in Pekin ducks there was a far greater loss of erythrocytes than could be accounted for as a result of direct physical rupture by the parasite. Erythrocyte loss began at the same time the 1st parasites appeared in the blood and was severest just prior to maximum parasitemia. Blood replacement and parasite loss occurred simultaneously. Examination of the spleen and bone marrow revealed that erythrophagocytosis was not the cause of anemia as reported for infections of Plasmodium, Babesia and Anaplasma. An anti-erythrocyte (A-E) factor was found in the serum of acutely infected ducks which agglutinated and hemolyzed normal untreated duck erythrocytes as well as infected cells. This A-E factor appeared when the 1st red cell loss was detected and reached its maximum titer just prior to the greatest red cell loss. Titers of the A-E factor were determined using normal uninfected erythrocytes at temperatures between 4 and 42 C. Cells agglutinated below 25 C and hemolyzed at 37 and 42 C. These results indicated that the A-E factor could be responsible for loss of cells other than those which were infected and could thus produce an excess loss of red cells. Attempts to implicate the A-E factor as an autoantibody were all negative. The A-E factor was present in the gamma fraction of acute serum but no anamnestic response could be detected when recovered ducks were reinfected. Anemia was never as severe in reinfections as in primary infections. The A-E factor also never reached as high a titer and was removed from the circulation very rapidly in reinfected ducks. It is concluded that red cell loss in ducks with acute Leucocytozoon disease results from intravascular hemolysis rather than erythrophagocytosis. The A-E factor responsible for hemolysis is more likely a parasite product rather than autoantibody.

  1. Agglutinating mouse IgG3 compares favourably with IgMs in typing of the blood group B antigen: Functionality and stability studies

    PubMed Central

    Klaus, Tomasz; Bzowska, Monika; Kulesza, Małgorzata; Kabat, Agnieszka Martyna; Jemioła-Rzemińska, Małgorzata; Czaplicki, Dominik; Makuch, Krzysztof; Jucha, Jarosław; Karabasz, Alicja; Bereta, Joanna

    2016-01-01

    Mouse immunoglobulins M (IgMs) that recognize human blood group antigens induce haemagglutination and are used worldwide for diagnostic blood typing. Contrary to the current belief that IgGs are too small to simultaneously bind antigens on two different erythrocytes, we obtained agglutinating mouse IgG3 that recognized antigen B of the human ABO blood group system. Mouse IgG3 is an intriguing isotype that has the ability to form Fc-dependent oligomers. However, F(ab′)2 fragments of the IgG3 were sufficient to agglutinate type B red blood cells; therefore, IgG3-triggered agglutination did not require oligomerization. Molecular modelling indicated that mouse IgG3 has a larger range of Fab arms than other mouse IgG subclasses and that the unique properties of mouse IgG3 are likely due to the structure of its hinge region. With a focus on applications in diagnostics, we compared the stability of IgG3 and two IgMs in formulated blood typing reagents using an accelerated storage approach and differential scanning calorimetry. IgG3 was much more stable than IgMs. Interestingly, the rapid decrease in IgM activity was caused by aggregation of the molecules and a previously unknown posttranslational proteolytic processing of the μ heavy chain. Our data point to mouse IgG3 as a potent diagnostic tool. PMID:27484487

  2. Agglutinating mouse IgG3 compares favourably with IgMs in typing of the blood group B antigen: Functionality and stability studies.

    PubMed

    Klaus, Tomasz; Bzowska, Monika; Kulesza, Małgorzata; Kabat, Agnieszka Martyna; Jemioła-Rzemińska, Małgorzata; Czaplicki, Dominik; Makuch, Krzysztof; Jucha, Jarosław; Karabasz, Alicja; Bereta, Joanna

    2016-08-03

    Mouse immunoglobulins M (IgMs) that recognize human blood group antigens induce haemagglutination and are used worldwide for diagnostic blood typing. Contrary to the current belief that IgGs are too small to simultaneously bind antigens on two different erythrocytes, we obtained agglutinating mouse IgG3 that recognized antigen B of the human ABO blood group system. Mouse IgG3 is an intriguing isotype that has the ability to form Fc-dependent oligomers. However, F(ab')2 fragments of the IgG3 were sufficient to agglutinate type B red blood cells; therefore, IgG3-triggered agglutination did not require oligomerization. Molecular modelling indicated that mouse IgG3 has a larger range of Fab arms than other mouse IgG subclasses and that the unique properties of mouse IgG3 are likely due to the structure of its hinge region. With a focus on applications in diagnostics, we compared the stability of IgG3 and two IgMs in formulated blood typing reagents using an accelerated storage approach and differential scanning calorimetry. IgG3 was much more stable than IgMs. Interestingly, the rapid decrease in IgM activity was caused by aggregation of the molecules and a previously unknown posttranslational proteolytic processing of the μ heavy chain. Our data point to mouse IgG3 as a potent diagnostic tool.

  3. INACTIVATION OF SEXUAL AGGLUTINATION IN HANSENULA WINGEI AND SACCHAROMYCES KLUYVERI BY DISULFIDE-CLEAVING AGENTS.

    PubMed

    TAYLOR, N W

    1964-10-01

    Taylor, Neil W. (Northern Regional Research Laboratory, Peoria, Ill.). Inactivation of sexual agglutination in Hansenula wingei and Saccharomyces kluyveri by disulfide-cleaving agents. J. Bacteriol. 88:929-936. 1964.-Mating types of both Hansenula wingei and Saccharomyces kluyveri can be activated to produce uniformly strong sexual agglutination by treatments with various solvents, such as 8 m LiBr. The strongly agglutinative mating-type preparations were irreversibly inactivated for sexual agglutination by various chemical treatments. Type 5 of H. wingei was inactivated by disulfide-cleaving reagents, but type 21 of H. wingei was not. Type 3 of S. kluyveri was more sensitive than type 26 of S. kluyveri to inactivation by disulfide-cleaving reagents. Comparison of sensitivities to these and other treatments, plus a moderately strong cross-agglutination between type 3 and type 21, indicated that the sexually agglutinative elements on type 3 are similar to type 5, and those of type 21 are similar to those of type 26. Inactivation-rate experiments showed a loss of agglutinative ability according to a sigmoid decrement with time for both types 5 and 21. The apparent extent of inactivation depended markedly on agglutination test conditions. Results of these experiments were interpreted to indicate tentatively, first, that the agglutinative elements of both types of a species are proteins and, second, that several agglutinating linkages are formed between any two cells in sexual agglutination.

  4. Rabbit hematology.

    PubMed

    Marshall, Kemba L

    2008-09-01

    Using laboratory animal medicine as an established resource, companion animal veterinarians have access to many physiologic and basic science studies that we can now merge with our clinical impressions. By working with reference laboratories, companion animal veterinarians are poised to accelerate our knowledge of the normal rabbit rapidly. The aim of this article is to discuss normal hematopoiesis and infectious and metabolic diseases that specifically target the hemolymphatic system. Additionally, photographic representation of cell types is provided.

  5. 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.

  6. Measurement of RBC agglutination with microscopic cell image analysis in a microchannel chip.

    PubMed

    Cho, Chi Hyun; Kim, Ju Yeon; Nyeck, Agnes E; Lim, Chae Seung; Hur, Dae Sung; Chung, Chanil; Chang, Jun Keun; An, Seong Soo A; Shin, Sehyun

    2014-01-01

    Since Landsteiner's discovery of ABO blood groups, RBC agglutination has been one of the most important immunohematologic techniques for ABO and RhD blood groupings. The conventional RBC agglutination grading system for RhD blood typings relies on macroscopic reading, followed by the assignment of a grade ranging from (-) to (4+) to the degree of red blood cells clumping. However, with the new scoring method introduced in this report, microscopically captured cell images of agglutinated RBCs, placed in a microchannel chip, are used for analysis. Indeed, the cell images' pixel number first allows the differentiation of agglutinated and non-agglutinated red blood cells. Finally, the ratio of agglutinated RBCs per total RBC counts (CRAT) from 90 captured images is then calculated. During the trial, it was observed that the agglutinated group's CRAT was significantly higher (3.77-0.003) than that of the normal control (0). Based on these facts, it was established that the microchannel method was more suitable for the discrimination between agglutinated RBCs and non-agglutinated RhD negative, and thus more reliable for the grading of RBCs agglutination than the conventional method.

  7. RABBIT POX

    PubMed Central

    Rosahn, Paul D.; Hu, Ch'uan-K'uei

    1935-01-01

    Observations on an epidemic of rabbit pox occurring in an isolated animal room during the winter of 1933–34 are reported. The clinical manifestations, consisting of a generalized papular eruption involving the skin and mucous membranes, together with blepharitis, ophthalmia, nasal discharge and lymphadenopathy were essentially similar to those noted in a pox epidemic of the previous year. This was true in general also of the pathological findings except that vacuolization, local necrosis and vesicle formation were seen in the epidermis, while in the previous year the microscopic pathology in the skin was confined to the corium. Evidence was presented indicating that the infection can be transmitted through the medium of a personal carrier, and that transmission in this manner can occur during the incubation period or before a definite diagnosis is possible. The findings also demonstrated that the etiological agents responsible for the disease reported here and that of the previous year were immunologically related, and that the immunity in recovered animals effectively persisted during the entire period for which data are available, 9 to 12 months. It appeared also that young animals suckling an immune doe were more refractory to the development of the lesions of rabbit pox than were the young of susceptible does. PMID:19870418

  8. Serogrouping of Bacteroides fragilis subsp. fragilis by the agglutination test.

    PubMed Central

    Lambe, D W; Moroz, D A

    1976-01-01

    The agglutination technique was used to establish a serological classification scheme for 98 strains of Bacteroides fragilis subsp. fragilis isolated from clinical specimens and normal human feces. Absorbed antisera were prepared to seven strains of B. fragilis subsp. fragilis. These seven absorbed antisera were species as well as subspecies specific and provided the basis of the serological classification scheme. This scheme was composed of 21 serogroups; seven of these serogroups contained only one group component. There was a total of 45 serological patterns. This serological scheme may be used for the serological classification of strains of B. fragilis subsp. fragilis and to study the epidemiology of this organism. PMID:950378

  9. Latex agglutination test (LAT) for the diagnosis of typhoid fever.

    PubMed

    Sahni, Gopal Shankar

    2013-06-01

    The efficacy of latex agglutination test in the rapid diagnosis of typhoid fever was studied and the result compared with that of blood culture. This study included 80 children suffering from typhoid fever, among which 40 were confirmed by blood culture isolation and 40 had possible typhoid fever based on high Widal's titre (a four-fold rise in the titre of antibody to typhi "O" and "H" antigen was considered as a positive Widal's test result). Eighty children, 40 with febrile illness confirmed to be other than typhoid and 40 normal healthy children were used as negative controls. The various groups were: (i) Study group ie, group I had 40 children confirmed by culture isolation of Salmonella typhi(confirmed typhoid cases). (ii) Control groups ie, (a) group II with 40 febrile controls selected from paediatrics ward where cause other than S typhi has been established, (b) group III with 40 afebrile healthy controls that were siblings of the children admitted in paediatric ward for any reason with no history of fever and TAB vaccination in the last one year, and (c) group IV with 40 children with high Widal's titre in paired sera sample. Widal's test with paired sera with a one week interval between collections were done in all 40 patients. Latex aggtutination test which could detect 900 ng/ml of antigen as observed in checker board titration, was positive in all 40 children from group I who had positive blood culture and in 30 children from group IV who had culture negative and had high Widal's titre positive. Latex agglutination test was positive in 4 children in group II and none in group III. Using blood culture positive cases as true positive and children in groups II and III as true negative, the test had a sensitivity of 100% and specificity of 96%. Latex agglutination test was found to be significantly sensitive (100%) and specific (96%) and could detect 75% more cases in group IV (possible typhoid cases). Thus latex agglutination test can be used for rapid

  10. Formation of agglutinate-like particles in an experimental regolith

    NASA Technical Reports Server (NTRS)

    See, Thomas H.; Horz, Friedrich

    1988-01-01

    Agglutinate-like particles composed predominantly of glass were produced from a fragmental gabbro target that was repetitively impacted by Ni-alloy projectiles. The experimental glasses are much more heterogeneous in composition than their lunar counterparts, and they are dominated by incomplete mixing of melted component minerals and by plagioclase-rich compositions. Most of the particles are found to be highly enriched in feldspar and to be sustantially fractionated relative to the initial bulk target. It is suggested that fractionation trends within lunar agglutinitic glasses may be partly due to phase-specific melting.

  11. The effect of subarachnoid erythrocyte lysate on brain injury: a preliminary study

    PubMed Central

    Zhang, Zi-Huan; Han, Yan-Ling; Wang, Chun-Xi; Zhou, Chen-Hui; Wu, Ling-Yun; Zhang, Hua-Sheng; Chen, Qiang; Fan, Jie-Mei; Zhou, Meng-Liang; Hang, Chun-Hua

    2016-01-01

    Abundant erythrocytes remain and lyse partially in the subarachnoid space after severe subarachnoid haemorrhage (SAH). But the effect of subarachnoid erythrocyte lysate on brain injury is still not completely clear. In this study, autologous erythrocytes (the non-lysate group) and their lysate (the lysate group) were injected separately into the cistern magna of rabbits to induce a model of experimental SAH, although the control group received isotonic sodium chloride solution instead of erythrocyte solution. Results showed that vasospasm of the basilar artery was observed at 72 h after experimental SAH, but there was no significant difference between the non-lysate group and the lysate group. Brain injury was more severe in the lysate group than in the non-lysate group. Meanwhile, the levels of peroxiredoxin 2 (Prx2), IL-6 and TNF-α in brain cortex and in CSF were significantly higher in the lysate group than those in the non-lysate group. These results demonstrated that brain injury was more likely to be caused by erythrocyte lysate than by intact erythrocytes in subarachnoid space, and inflammation response positively correlated with Prx2 expression might be involved in mechanism of brain injury after SAH. PMID:27279653

  12. Comparative study of erythrocyte glycolytic enzymes in man and in two species of Lagomorphae. (Ochotona rufescens rufescens and Oryctolagus cuniculus).

    PubMed

    Puget, A; Vergnes, H; Gouarderes, C

    1974-07-01

    The authors made a comparative study of red cell enzyme glycolysis in man and in two species of Lagomorphae, the pika and the rabbit. The activities of the 12 enzymes of Embden-Meyerhoff pathway and of the two dehydrogenases of pentose shunt (glucose-6-phosphate dehydrogenase and 6-phosphogluconate dehydrogenase) were determined. Phosphoglycerate kinase and pyruvate kinase showed quite similar activities in pika erythrocytes and in erythrocytes from human umbilical cord. The levels of these enzymes differed significantly in the pika and in the rabbit. No differences were noted between pyruvate kinases from the rabbit and the neonatal man. The other activities gave values either identical to those found in the adult man or intermediate between the adult and the neonatal man. In the rabbit the levels of glycolitic enzymes were generally lower than in the pika except for lactate dehydrogenase and glycose-6-phosphate dehydrogenase. Some characteristics of erythrocyte metabolism specific to the pika may account for the differences observed in this species. The influence of red cell age cannot explain the variations observed for no significant reticulocytosis was observed in the circulating blood. The percentages found in the pika and the rabbit were essentially identical at determination.

  13. Quantitative determination of fibrinogen of patients with coronary heart diseases through piezoelectric agglutination sensor.

    PubMed

    Chen, Qinghai; Hua, Xing; Fu, Weiling; Liu, Dongbo; Chen, Ming; Cai, Guoru

    2010-01-01

    Fibrinogen can transform fibrin through an agglutination reaction, finally forming fibrin polymer with grid structure. The density and viscosity of the reaction system changes drastically during the course of agglutination. In this research, we apply an independently-developed piezoelectric agglutination sensor to detect the fibrinogen agglutination reaction in patients with coronary heart diseases. The terminal judgment method of determining plasma agglutination reaction through piezoelectric agglutination sensor was established. In addition, the standard curve between plasma agglutination time and fibrinogen concentration was established to determinate fibrinogen content quantitatively. The results indicate the close correlation between the STAGO paramagnetic particle method and the method of piezoelectric agglutination sensor for the detection of Fibrinogen. The correlation coefficient was 0.91 (γ = 0.91). The determination can be completed within 10 minutes. The fibrinogen concentration in the coronary heart disease group was significantly higher than that of the healthy control group (P < 0.05). The results reveal that high fibrinogen concentration is closely correlated to the incurrence, development and prognosis of coronary heart diseases. Compared with other traditional methods, the method of piezoelectric agglutination sensor has some merits such as operation convenience, small size, low cost, quick detecting, good precision and the common reacting agents with paramagnetic particle method.

  14. Ultrasensitive Antibody Detection by Agglutination-PCR (ADAP)

    PubMed Central

    2016-01-01

    Antibodies are widely used biomarkers for the diagnosis of many diseases. Assays based on solid-phase immobilization of antigens comprise the majority of clinical platforms for antibody detection, but can be undermined by antigen denaturation and epitope masking. These technological hurdles are especially troublesome in detecting antibodies that bind nonlinear or conformational epitopes, such as anti-insulin antibodies in type 1 diabetes patients and anti-thyroglobulin antibodies associated with thyroid cancers. Radioimmunoassay remains the gold standard for these challenging antibody biomarkers, but the limited multiplexability and reliance on hazardous radioactive reagents have prevented their use outside specialized testing facilities. Here we present an ultrasensitive solution-phase method for detecting antibodies, termed antibody detection by agglutination-PCR (ADAP). Antibodies bind to and agglutinate synthetic antigen–DNA conjugates, enabling ligation of the DNA strands and subsequent quantification by qPCR. ADAP detects zepto- to attomoles of antibodies in 2 μL of sample with a dynamic range spanning 5–6 orders of magnitude. Using ADAP, we detected anti-thyroglobulin autoantibodies from human patient plasma with a 1000-fold increased sensitivity over an FDA-approved radioimmunoassay. Finally, we demonstrate the multiplexability of ADAP by simultaneously detecting multiple antibodies in one experiment. ADAP’s combination of simplicity, sensitivity, broad dynamic range, multiplexability, and use of standard PCR protocols creates new opportunities for the discovery and detection of antibody biomarkers. PMID:27064772

  15. Hemagglutinating activity of serovar reference strains of Ornithobacterium rhinotracheale.

    PubMed

    Vega, Vicente; Zepeda, Andrea; Ramírez, Saúl; Morales, Vladimir; Fernández, Pomposo; de Oca, Roberto Montes; Guerra-Infante, Fernando M; de Jesús de Haro-Cruz, María; Blackall, Patrick J; Soriano, Edgardo V

    2008-05-01

    In the present study, the hemagglutinating activity of 9 reference strains (serovars A-I) of Ornithobacterium rhinotracheale was investigated by using fresh erythrocytes from 15 different species: chicken (broiler, rooster, hen), turkey, pigeon, quail, duck, Harris hawk (Parabuteo unicinctus), house finch (Carpodacus mexicanus), cow, sheep, horse, dog, rabbit, pig, human (groups A, B, AB, and O), and rainbow trout (Oncorhynchus mykiss). All 9 strains agglutinated rabbit erythrocytes. None of the strains was able to agglutinate hen, cow, horse, or rainbow trout erythrocytes. The number of positive reactions among the remaining species varied. Results indicate that the use of rabbit erythrocytes is better suited for testing the hemagglutinating activity of O. rhinotracheale.

  16. Viral infections of rabbits.

    PubMed

    Kerr, Peter J; Donnelly, Thomas M

    2013-05-01

    Viral diseases of rabbits have been used historically to study oncogenesis (e.g. rabbit fibroma virus, cottontail rabbit papillomavirus) and biologically to control feral rabbit populations (e.g. myxoma virus). However, clinicians seeing pet rabbits in North America infrequently encounter viral diseases although myxomatosis may be seen occasionally. The situation is different in Europe and Australia, where myxomatosis and rabbit hemorrhagic disease are endemic. Advances in epidemiology and virology have led to detection of other lapine viruses that are now recognized as agents of emerging infectious diseases. Rabbit caliciviruses, related to rabbit hemorrhagic disease, are generally avirulent, but lethal variants are being identified in Europe and North America. Enteric viruses including lapine rotavirus, rabbit enteric coronavirus and rabbit astrovirus are being acknowledged as contributors to the multifactorial enteritis complex of juvenile rabbits. Three avirulent leporid herpesviruses are found in domestic rabbits. A fourth highly pathogenic virus designated leporid herpesvirus 4 has been described in Canada and Alaska. This review considers viruses affecting rabbits by their clinical significance. Viruses of major and minor clinical significance are described, and viruses of laboratory significance are mentioned.

  17. A novel flow cytometry-based assay for the quantification of antibody-dependent pneumococcal agglutination

    PubMed Central

    van der Gaast—de Jongh, Christa E.; Diavatopoulos, Dimitri A.; de Jonge, Marien I.

    2017-01-01

    The respiratory pathogen Streptococcus pneumoniae is a major cause of diseases such as otitis media, pneumonia, sepsis and meningitis. The first step towards infection is colonization of the nasopharynx. Recently, it was shown that agglutinating antibodies play an important role in the prevention of mucosal colonization with S. pneumoniae. Here, we present a novel method to quantify antibody-dependent pneumococcal agglutination in a high-throughput manner using flow cytometry. We found that the concentration of agglutinating antibodies against pneumococcal capsule are directly correlated with changes in the size and complexity of bacterial aggregates, as measured by flow cytometry and confirmed by light microscopy. Using the increase in size, we determined the agglutination index. The cutoff value was set by measuring a series of non-agglutinating antibodies. With this method, we show that not only anti-polysaccharide capsule antibodies are able to induce agglutination but that also anti-PspA protein antibodies have agglutinating capabilities. In conclusion, we have described and validated a novel method to quantify pneumococcal agglutination, which can be used to screen sera from murine or human vaccination studies, in a high-throughput manner. PMID:28288168

  18. 9 CFR 147.1 - The standard tube agglutination test. 1

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... 9 Animals and Animal Products 1 2014-01-01 2014-01-01 false The standard tube agglutination test... Blood Testing Procedures § 147.1 The standard tube agglutination test. 1 1 The procedure described is a... containers provided by the laboratory. The containers should be stout-walled test tubes, preferably 3/8 by...

  19. 9 CFR 147.1 - The standard tube agglutination test. 1

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 9 Animals and Animal Products 1 2010-01-01 2010-01-01 false The standard tube agglutination test... Blood Testing Procedures § 147.1 The standard tube agglutination test. 1 1 The procedure described is a... containers provided by the laboratory. The containers should be stout-walled test tubes, preferably 3/8 by...

  20. 9 CFR 147.1 - The standard tube agglutination test. 1

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 9 Animals and Animal Products 1 2011-01-01 2011-01-01 false The standard tube agglutination test... Blood Testing Procedures § 147.1 The standard tube agglutination test. 1 1 The procedure described is a... containers provided by the laboratory. The containers should be stout-walled test tubes, preferably 3/8 by...

  1. [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.

  2. 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 ...

  3. 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.

  4. An early Cambrian agglutinated tubular lophophorate with brachiopod characters

    PubMed Central

    Zhang, Z.-F.; Li, G.-X.; Holmer, L. E.; Brock, G. A.; Balthasar, U.; Skovsted, C. B.; Fu, D.-J.; Zhang, X.-L.; Wang, H.-Z.; Butler, A.; Zhang, Z.-L.; Cao, C.-Q.; Han, J.; Liu, J.-N.; Shu, D.-G.

    2014-01-01

    The morphological disparity of lophotrochozoan phyla makes it difficult to predict the morphology of the last common ancestor. Only fossils of stem groups can help discover the morphological transitions that occurred along the roots of these phyla. Here, we describe a tubular fossil Yuganotheca elegans gen. et sp. nov. from the Cambrian (Stage 3) Chengjiang Lagerstätte (Yunnan, China) that exhibits an unusual combination of phoronid, brachiopod and tommotiid (Cambrian problematica) characters, notably a pair of agglutinated valves, enclosing a horseshoe-shaped lophophore, supported by a lower bipartite tubular attachment structure with a long pedicle with coelomic space. The terminal bulb of the pedicle provided anchorage in soft sediment. The discovery has important implications for the early evolution of lophotrochozoans, suggesting rooting of brachiopods into the sessile lophotrochozoans and the origination of their bivalved bauplan preceding the biomineralization of shell valves in crown brachiopods. PMID:24828016

  5. A rapid latex agglutination test for detection of leptospiral antibodies.

    PubMed

    Ramadass, P; Samuel, B; Nachimuthu, K

    1999-10-01

    A rapid semi-quantitative latex agglutination test (LAT) has been standardized for the detection of leptospiral antibodies in serum samples of man and animals. The efficacy of the LAT was compared with the plate enzyme linked immunosorbent assay (ELISA). A total of 276 human serum samples were analyzed by both LAT and ELISA and percentage positives were 84.8 and 85.9%, respectively. Similarly, of 65 animal samples tested, 63.1 and 69.2% positivity were observed in LAT and ELISA, respectively. Even though the ELISA test was slightly more sensitive than LAT, the rapidity, simplicity and economics of the LAT were found to fulfill the requirements of a screening test for leptospiral antibodies.

  6. Latex agglutination: diagnose the early cryptococcus neoformans test of capsular polysaccharide antigen.

    PubMed

    Wang, Huanrong; Yuan, Xueqian; Zhang, Lifeng

    2015-01-01

    This paper aims to discuss the early diagnosis value of latex agglutination test in Cryptococcal meningitis. The cerebrospinal fluid (CSF) of 112 patients with definite Cryptococcal meningitis and 26 patients with tubercular meningitis and virus meningitis were collected, latex agglutination test is adopted to detect Cryptococcal capsular polysaccharide antigen. Then it was compared with fungal culture and direct microscopy method for evaluating the sensitivity and specificity of the diagnosis. The sensitivity of three methods including latex agglutination test, fungal culture and direct microscopy was 91.1%,69.6% and 73.2% respectively. The specificity of latex agglutination test was 96.0%, 100% and 100% respectively. That latex agglutination test to detect Cryptococcal capsular polysaccharide antigen could be taken as the early diagnostic method of Cryptococcus neoformans meningitis.

  7. Sperm-agglutinating antibodies and testicular morphology in fifty-nine men with azoospermia or cryptozoospermia.

    PubMed

    Friberg, J; Kjessler, B

    1975-04-01

    The relationship between the state of the germinal epithelium and the type and titer of circulating sperm-agglutinating antibodies has been investigated in a series of 59 azoospermic or occasionally cryptozoospermic men. The patients were grouped according to the condition of the germinal epithelium as observed from testicular biopsy specimens, as well as to type and titer of circulating sperm-agglutinating antibodies investigated by a previously described microagglutination technique. Evidence is presented to suggest that the presence of mature spermatozoa in the testicular structures may be a prerequisite for the spontaneous production of circulating sperm-agglutinating antibodies, at least of the head-to-tail (H-T) agglutinating type. Furthermore, these circulating H-T sperm-agglutinating antibodies, once they are formed, do not seem to interfere adversely with the germinal epithelium of the carrier.

  8. Macroscopic Agglutination Test for Rapid Diagnosis of Human Leptospirosis

    PubMed Central

    Brandão, Angela P.; Camargo, Eide D.; da Silva, Emilson D.; Silva, Marcos V.; Abrão, Rui V.

    1998-01-01

    A commercially available slide agglutination test (SAT) for the diagnosis of human leptospirosis was evaluated by comparing it to an immunoglobulin M (IgM) enzyme-linked immunosorbent assay (ELISA) and to the microscopic agglutination test (MAT). For all 108 patients, leptospirosis was diagnosed on the basis of a fourfold or greater increase in titer by MAT (seroconversion), and all but 1 of 245 controls were MAT negative (titers, <1:100). Both SAT and the IgM ELISA failed to detect one case of infection (sensitivity, 99%). Only 3 of 145 blood donors and none of the 100 patients with other illnesses were SAT positive (specificity, 99%). The overall results were similar for the three tests; however, SAT and ELISA were statistically more sensitive as initial screening tests. For 22% of the patients, the diagnosis of leptospirosis was made earlier by SAT than by MAT. SAT detected 27 (44%) of 62 MAT-negative patients with the first serum sample. ELISA and SAT had very similar results. Follow-up of patients for 1 year after the onset of symptoms showed a decreasing rate of positivity by SAT from the third month on. The rate of positivity by ELISA decreased more slowly, to about 67% by the end of the study. By MAT all patients were persistently reactive. SAT and ELISA seem to be convenient methods for the rapid and early screening for leptospirosis and could replace the less sensitive MAT. ELISA gives less subjective results than SAT and provides information on IgM kinetics, but it can be performed only by the more sophisticated laboratories. SAT is inexpensive, can be performed more quickly and more easily than ELISA, and could be used by the less well equipped laboratories. PMID:9774553

  9. 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…

  10. [Insulin and glucocorticoid binding by blood cell receptors after hydrocortisone administration in rabbits].

    PubMed

    Tikhonova, N E; Tatarinova, G Sh

    1989-07-01

    Repeated i.v. administration of hydrocortisone (10 mg/kg) revealed an increase in the resistance against insulin although endogenous corticosterone was decreased in 33 male rabbits. The insulin- and dexamethasone-binding receptors of erythrocytes and mononuclear leucocytes. changed after 3-7 hydrocortisone injections, the binding increasing for insulin and diminishing for dexamethasone.

  11. 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.

  12. Superoxide dismutase activity in radioresistant tissues of irradiated rabbits.

    PubMed

    Stoklasová, A; Kovárová, H; Ledvina, M

    1992-01-01

    The activities of Cu, Zn-containing superoxide dismutase were studied in radioresistant tissues (liver, brain, erythrocytes) of whole-body irradiated rabbits with 6.0 Gy and 24.0 Gy with local shielding. No significant changes were observed after irradiation with 6.0 Gy. Both the changes in Cu, Zn-SOD activity and the protein concentrations were more pronounced after exposure to 24.0 Gy with local shielding of the head and abdominal region. The dose on the shielded regions was about 6.0 Gy. Local shielding of rabbits irradiated with a lethal dose 24.0 Gy influenced positively the survival of animals. However, the decrease in SOD activity on 60th day after irradiation seems to be unfavourable for further survival of rabbits, if we accept that SOD content in tissue is maintained at a rather constant level.

  13. Erythrocytes from GGTA1/CMAH knockout pigs: implications for xenotransfusion and testing in non-human primates

    PubMed Central

    Wang, Zheng-Yu; Burlak, Christopher; Estrada, Jose L.; Li, Ping; Tector, Matthew F.; Tector, A. Joseph

    2015-01-01

    Background Pig erythrocytes are potentially useful to solve the worldwide shortage of human blood for transfusion. Domestic pig erythrocytes, however, express antigens that are bound by human preformed antibodies. Advances in genetic engineering have made it possible to rapidly knock out the genes of multiple xenoantigens, namely galactose α1,3 galactose (aGal) and N-glycolylneuraminic acid (Neu5Gc). We have recently targeted the GGTA1 and CMAH genes with zinc finger endonucleases resulting in double knockout pigs that no longer express aGal or Neu5Gc and attract significantly fewer human antibodies. In this study, we characterized erythrocytes from domestic and genetically modified pigs, baboons, chimpanzees, and humans for binding of human and baboon natural antibody, and complement mediated lysis. Methods Distribution of anti Neu5Gc IgG and IgM in pooled human AB serum was analyzed by ELISA. Erythrocytes from domestic pigs (Dom), aGal knockout pigs (GGTA1 KO), aGal and Neu5Gc double knockout pigs (GGTA1/CMAH KO), baboons, chimpanzees, and humans were analyzed by flow cytometry for aGal and Neu5Gc expression. In vitro comparative analysis of erythrocytes was conducted with pooled human AB serum and baboon serum. Total antibody binding was accessed by hemagglutination; complement-dependent lysis was measured by hemolytic assay; IgG or IgM binding to erythrocytes was characterized by flow cytometry. Results The pooled human AB serum contained 0.38 μg/ml anti Neu5Gc IgG and 0.085 μg/ml anti Neu5Gc IgM. Both Gal and Neu5Gc were not detectable on GGTA1/CMAH KO erythrocytes. Hemagglutinaion of GGTA1/CMAH KO erythrocytes with human serum was 3.5-fold lower compared to GGTA1 KO erythrocytes, but 1.6-fold greater when agglutinated with baboon serum. Hemolysis of GGTA1/CMAH KO erythrocytes by human serum (25%) was reduced 9-fold compared to GGTA1 KO erythrocytes, but increased 1.64-fold by baboon serum. Human IgG binding was reduced 27-fold on GGTA1/CMAH KO erythrocytes

  14. Variant rabbit hemorrhagic disease virus in young rabbits, Spain.

    PubMed

    Dalton, Kevin P; Nicieza, Inés; Balseiro, Ana; Muguerza, María A; Rosell, Joan M; Casais, Rosa; Álvarez, Ángel L; Parra, Francisco

    2012-12-01

    Outbreaks of rabbit hemorrhagic disease have occurred recently in young rabbits on farms on the Iberian Peninsula where rabbits were previously vaccinated. Investigation identified a rabbit hemorrhagic disease virus variant genetically related to apathogenic rabbit caliciviruses. Improved antivirus strategies are needed to slow the spread of this pathogen.

  15. Plasmodium vivax Invasion of Human Erythrocytes Inhibited by Antibodies Directed against the Duffy Binding Protein

    PubMed Central

    Grimberg, Brian T; Udomsangpetch, Rachanee; Xainli, Jia; McHenry, Amy; Panichakul, Tasanee; Sattabongkot, Jetsumon; Cui, Liwang; Bockarie, Moses; Chitnis, Chetan; Adams, John; Zimmerman, Peter A; King, Christopher L

    2007-01-01

    Background Plasmodium vivax invasion requires interaction between the human Duffy antigen on the surface of erythrocytes and the P. vivax Duffy binding protein (PvDBP) expressed by the parasite. Given that Duffy-negative individuals are resistant and that Duffy-negative heterozygotes show reduced susceptibility to blood-stage infection, we hypothesized that antibodies directed against region two of P. vivax Duffy binding protein (PvDBPII) would inhibit P. vivax invasion of human erythrocytes. Methods and Findings Using a recombinant region two of the P. vivax Duffy binding protein (rPvDBPII), polyclonal antibodies were generated from immunized rabbits and affinity purified from the pooled sera of 14 P. vivax–exposed Papua New Guineans. It was determined by ELISA and by flow cytometry, respectively, that both rabbit and human antibodies inhibited binding of rPvDBPII to the Duffy antigen N-terminal region and to Duffy-positive human erythrocytes. Additionally, using immunofluorescent microscopy, the antibodies were shown to attach to native PvDBP on the apical end of the P. vivax merozoite. In vitro invasion assays, using blood isolates from individuals in the Mae Sot district of Thailand, showed that addition of rabbit anti-PvDBPII Ab or serum (antibodies against, or serum containing antibodies against, region two of the Plasmodium vivax Duffy binding protein) (1:100) reduced the number of parasite invasions by up to 64%, while pooled PvDBPII antisera from P. vivax–exposed people reduced P. vivax invasion by up to 54%. Conclusions These results show, for what we believe to be the first time, that both rabbit and human antibodies directed against PvDBPII reduce invasion efficiency of wild P. vivax isolated from infected patients, and suggest that a PvDBP-based vaccine may reduce human blood-stage P. vivax infection. PMID:18092885

  16. Momordica charantia seed lectin: toxicity, bacterial agglutination and antitumor properties.

    PubMed

    Kabir, Syed Rashel; Nabi, Md Mahamodun; Nurujjaman, Md; Abu Reza, Md; Alam, A H M Khurshid; Uz Zaman, Rokon; Khalid-Bin-Ferdaus, Khandaker Md; Amin, Ruhul; Khan, Md Masudul Hasan; Hossain, Md Anowar; Uddin, Md Salim; Mahmud, Zahid Hayat

    2015-03-01

    In last three decades, several studies were carried out on the D-galactose-specific lectin of Momordica charantia seeds (MCL). In the present study, in vitro growth inhibition (8-23 %) at different concentrations (6-24 μg/ml) of MCL was observed against Ehrlich ascites carcinoma (EAC) cells by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. MCL also showed 28, 45, and 75 % growth inhibitions against EAC cells when administered 1.2, 2.0, and 2.8 mg/kg/day (i.p.), respectively for five consequent days in vivo in mice. After lectin treatment, the level of red blood cell and hemoglobin was increased significantly with the decrease of white blood cell and maintained the normal level when compared with EAC-bearing control and normal mice without EAC cells. Although MCL caused cell cycle arrest at G0/G1 phase of EAC cells, any irregular shape or apoptotic morphological alterations in the lectin-treated EAC cells was not observed by an optical and fluorescence microscope. Lectin showed toxicity against brine shrimp nauplii with an LC50 value of 49.7 μg/ml. Four out of seven pathogenic bacteria were agglutinated by MCL in the absence of inhibitory sugar D-lactose/D-galactose. In conclusion, MCL showed strong cytotoxic effect and therefore can be used as a potent anticancer chemotherapeutic agent.

  17. Prolonged oral cyanide effects on feed intake, growth rate and blood parameters in rabbits.

    PubMed

    Avais, Muhammad; Khan, Muhammad Sarwar; Khan, Muhammad Arif; Ashraf, Kamran; Khan, Jawaria Ali; Hameed, Sajid

    2014-07-01

    Twelve adult rabbits bred locally were divided into two equal groups of 6; experimental and control groups. Rabbits in the experimental group were orally dosed with KCN at 3mg/kg body weight for 40 consecutive days. Members in control group were given placebo (distilled water) for the same period. Animals in both groups were offered feed at 90gm/kg/day while ample drinking water was available ad lib. Feed consumption and body weight of rabbits in both the groups were recorded. Blood samples were also drawn to determine various hematological parameters. Statistical analysis revealed a non-significant difference of total and daily feed intakes in rabbits of experimental and control groups. Whereas the feed efficiency of rabbits in the experimental group were significantly reduced (P<0.05) compared to controls. Likewise a significant decrease in body weight gain of rabbits in experimental group (P<0.05) was observed. A non-significant difference (P>0.05) was observed in leukocyte count, differential leukocyte count and platelets of rabbits in both the groups. Erythrocyte count, hemoglobin concentration, packed cell volume and mean corpuscular hemoglobin were significantly decreased in treated rabbits. It was concluded that chronic cyanide intake had a deleterious effect on feed efficiency, growth rate and blood components of rabbits.

  18. 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.

  19. The Cenozoic Diversity of Agglutinated Foraminifera - Evidence for a late Oligocene to early Miocene diversification event

    NASA Astrophysics Data System (ADS)

    Kaminski, Michael; Setoyama, Eiichi; Kender, Sev; Cetean, Claudia

    2014-05-01

    The agglutinated foraminifera are among the most abundant micro-organisms in the deep marine environment and have a diversity record extending back to the late Precambrian. We present an updated diversity curve for agglutinated foraminiferal genera based on the stratigraphic ranges of all the agglutinated genera recognized as valid in the classification of Kaminski (2014). The data set for this analysis is based on the stratigraphic ranges of agglutinated genera published in Foraminiferal Genera and their Classification, which has been subsequently updated based on published studies and our new observations. The mean standing diversity of agglutinated foraminiferal genera was compiled by counting the number of boundary crossers rather than the number of genera in each stage. In this study, we report the stratigraphic and geographical occurrence of a benthic foraminiferal diversification event that has previously received little attention. In the latest Oligocene to earliest Miocene a number of trochospiral agglutinated genera with alveolar or canaliculate walls first appeared in the fossil record. Our studies of late Oligocene of the Congo fan, offshore Angola (Kender et al., 2008; Cetean and Kaminski, 2011) have revealed a diverse assemblage that includes new taxa of deep-water agglutinated foraminifera. In a biostratigraphic study of the Miocene foraminiferal assemblages Kender et al. (2008) noted steadily increasing diversity and proportions of infaunal agglutinated foraminiferal morphotypes over the lower Miocene interval. The proportion of infaunal agglutinated foraminifera assigned to the order Textularida increased dramatically in the lower mid-Miocene, suggesting expansion of the oxygen minimum zone into deeper waters. In addition to the trochospiral alveolar genera, several species of Reticulophragmium and Cyclammina display rapid diversification into numerous separate lineages that are at present not reflected in our generic diversity record owing to

  20. [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.

  1. Modulation of ligand-mediated human red cell agglutinability by prostaglandins

    SciTech Connect

    McLawhon, R.W.; Marikovsky, Y.; Weinstein, R.S.

    1986-03-01

    Ethanol induces the transformation of human red cells from bioconcave discs to echinocytes in vitro. In addition, they have observed that ethanol can enhance the agglutination of red cells by the plant lectin wheat germ agglutinin or poly-L-lysine. Incubation of washed human red cells with 5 and 10% ethanol (v/v) in phosphate buffered saline, pH 7.3 at 25/sup 0/C produced a 30% increase in ligand-mediated agglutinability within 12 min. Simultaneous addition of ethanol and one of the following prostaglandin derivatives, PGE/sub 1/, pge/sub 2/, pgf/sub 2/-alpha, or PGl/sub 2/ (10/sup -9/ to 5 x 10/sup -7/ M) prevented the shape-associated increases in red cell agglutinability. Thromboxane-B/sub 2/ had no effect on agglutinability. Prostaglandins did not prevent ethanol-induced red cell shape transformations per se under identical experimental conditions. As intragastric administration of 100% ethanol results in the formation of spiculated red cell thrombi in postcapillary venules of rat gastric mucosa, they postulate that the cytoprotective role of prostanoids in preventing mucosal ulceration may be due in part to their capacity to inhibit intravascular ligand mediated red cell agglutination, hemostasis, and their sequelae, epithelial necrosis. Moreover, the data suggest that ethanol-induced red cell shape transformations and ligand-mediated agglutination represent two distinct and independent biological phenomena.

  2. Agglutination by anti-capsular polysaccharide antibody is associated with protection against experimental human pneumococcal carriage

    PubMed Central

    Reiné, J; Zangari, T; Owugha, JT; Pennington, SH; Gritzfeld, JF; Wright, AD; Collins, AM; van Selm, S; de Jonge, MI; Gordon, SB; Weiser, JN; Ferreira, DM

    2016-01-01

    The ability of pneumococcal conjugate vaccine (PCV) to decrease transmission by blocking the acquisition of colonization has been attributed to herd immunity. We describe the role of mucosal IgG to capsular polysaccharide (CPS) in mediating protection from carriage, translating our findings from a murine model to humans. We used a flow-cytometric assay to quantify antibody-mediated agglutination demonstrating that hyperimmune sera generated against an unencapsulated mutant was poorly agglutinating. Passive immunization with this antiserum was ineffective to block acquisition of colonization compared to agglutinating antisera raised against the encapsulated parent strain. In the human challenge model samples were collected from PCV and control vaccinated adults. In PCV-vaccinated subjects IgG levels to CPS were increased in serum and nasal wash (NW). IgG to the inoculated strain CPS dropped in NW samples after inoculation suggesting its sequestration by colonizing pneumococci. In post-vaccination NW samples pneumococci were heavily agglutinated compared to pre-vaccination samples in subjects protected against carriage. Our results indicate that pneumococcal agglutination mediated by CPS specific antibodies is a key mechanism of protection against acquisition of carriage. Capsule may be the only vaccine target that can elicit strong agglutinating antibody responses, leading to protection against carriage acquisition and generation of herd immunity. PMID:27579859

  3. A particle agglutination assay for rapid identification of heparin binding to coagulase-negative staphylococci.

    PubMed

    Pascu, C; Hirmo, S; Ljungh, A; Wadström, T

    1996-10-01

    The heparin-binding properties of six different species of coagulase-negative staphylococci were examined by a particle agglutination assay. Heparin (mol. wt 4000-6000), mildly treated with sodium periodate, was covalently coupled to amino-modified latex beads (0.72 micron diameter). The particle agglutination assay was validated by comparing results with the adhesion (percentage binding of adherent cells) of coagulase-negative staphylococcal strains to heparinised microtitration plates. Of 38 different coagulase-negative staphylococcal strains tested, 30 showed agglutination reactivity with heparin-coated latex beads. Strains of different coagulase-negative staphylococcal species agglutinated heparin-coated latex beads to various extents (e.g., cells of Staphylococcus haemolyticus strains reacted more strongly than cells of S. epidermidis strains). The agglutination reaction was significantly inhibited by fucoidan, suramin, lambda-carrageenan and other sulphated compounds, but not by non-sulphated carbohydrate polymers such as hyaluronic acid. Agglutination of staphylococcal cells with heparin-coated latex beads was completely blocked by a cell-surface extract. These results suggest that structures responsible for heparin binding are exposed on the cell surface.

  4. INFECTIOUS PAPILLOMATOSIS OF RABBITS

    PubMed Central

    Shope, Richard E.; Hurst, E. Weston

    1933-01-01

    A papilloma has been observed in wild cottontail rabbits and has been found to be transmissible to both wild and domestic rabbits. The clinical and pathological pictures of the condition have been described. It has been found that the causative agent is readily filtrable through Berkefeld but not regularly through Seitz filters, that it stores well in glycerol, that it is still active after heating to 67°C. for 30 minutes, but not after heating to 70°C., and that it exhibits a marked tropism for cutaneous epithelium. The activities and properties of the papilloma-producing agent warrant its classification as a filtrable virus. Rabbits carrying experimentally produced papillomata are partially or completely immune to reinfection and, furthermore, their sera partially or completely neutralize the causative virus. The disease is transmissible in series through wild rabbits and virus of wild rabbit origin is readily transmissible to domestic rabbits, producing in this species papillomata identical in appearance with those found in wild rabbits. However, the condition is not transmissible in series through domestic rabbits. The possible significance of this observation has been discussed. The virus of infectious papillomatosis is not related immunologically to either the virus of infectious fibroma or to that of infectious myxoma of rabbits. PMID:19870219

  5. [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.

  6. 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.

  7. Inadequate dietary magnesium intake increases atherosclerotic plaque development in rabbits

    PubMed Central

    King, Jennifer L.; Miller, Rita J.; Blue, James P.; O'Brien, William D.; Erdman, John W.

    2012-01-01

    Epidemiological studies have shown dietary magnesium (Mg) intake and serum Mg levels to be inversely correlated with the development of atherosclerosis. We hypothesized that low levels of Mg would promote atherosclerotic plaque development in rabbits. New Zealand white rabbits (4 months old, n = 22) were fed an atherogenic diet containing 0.12% (−Mg), 0.27% (control), or 0.43% (+Mg) Mg for 8 weeks. Blood samples were obtained at baseline, 2, 4, 6, and 8 weeks and were assayed for total cholesterol, high-density lipoprotein (HDL), non-HDL, triglycerides (TG), C-reactive protein, serum Mg, and erythrocyte Mg. Aortas from −Mg had significantly more plaque, with an intima thickness 42% greater than control and 36% greater than +Mg. Serum cholesterol levels rose over time, and at 8 weeks, −Mg had the highest and +Mg the lowest total and non-HDL cholesterol and TG levels, although these results did not reach significance. Over time, serum Mg levels increased, and erythrocyte Mg levels decreased. C-reactive protein significantly increased in all groups at 4 and 6 weeks but returned to baseline levels by 8 weeks. This study supports the hypothesis that inadequate intake of Mg results in an increase in atherosclerotic plaque development in rabbits. PMID:19555816

  8. Stability of Erythrocyte Ghosts: A γ -Ray Perturbed Angular Correlation Study

    NASA Astrophysics Data System (ADS)

    Kruse, Carol A.; Tin, George W.; Baldeschwieler, John D.

    1983-03-01

    The structural integrity of erythrocyte ghosts made by the preswell and slow-dialysis techniques has been studied in vitro by use of γ -ray perturbed angular correlation (PAC) techniques and also by standard in vitro leakage methods employing sequestered labeled markers. Complexes of 111In3+ and nitrilotriacetate were encapsulated in ghosts made from human, rabbit, rat, and mouse erythrocytes, and their leakage was monitored by both methods. In addition, 125I-labeled bovine serum albumin was encapsulated, and ghost integrity was monitored by conventional leakage measurements. With the PAC technique the percentage of material released from human ghosts was determined quantitatively, and the results were equivalent to those obtained by the conventional method. In addition, at various times after intravenous injection, tissue distribution of the ghosts in the mouse was studied. The percent injected dose per gram of tissue of the labeled surface proteins of erythrocyte ghosts in circulation approximated that of the entrapped labeled albumin. This suggests that the ghost membrane and contents are strongly associated in vivo. Large 125I-labeled bovine serum albumin molecules and small 111In3+-nitrilotriacetate complexes were delivered in high quantities to the lung initially, and to the liver and spleen. Because erythrocyte ghosts have the ability to entrap a wide range of substances and deliver them to specific organs, ghosts may be preferable to other drug carriers or drug therapy for treatment of certain disorders.

  9. Autoantibody Production in Rabbits

    PubMed Central

    Asherson, G. L.; Rose, M. Elaine

    1963-01-01

    The finding that the serum of apparently healthy rabbits fixed complement with rabbit liver and kidney has been confirmed. Experimental infection of rabbits with Eimeria stiedae, the cause of hepatic coccidiosis, led to a rise in the titre of serum complement-fixing factors. The rise was statistically significant 14, 21 and 28 days after infection. The factors were regarded as antibodies because they behaved as macroglobulins on diethylaminoethyl—cellulose chromatography and sucrose gradient centrifugation, and as autoantibodies because they fixed complement with the kidney of the rabbits in which they occurred. The antibody reacted with widely distributed antigen(s) with high activity in brain and low activity in skeletal muscle. The possibility that coccidial infection may be responsible for the natural autoantibody of rabbits is discussed. PMID:13965167

  10. Studies on the In Vivo Effects of Antibody INTERACTION OF IgM ANTIBODY AND COMPLEMENT IN THE IMMUNE CLEARANCE AND DESTRUCTION OF ERYTHROCYTES IN MAN

    PubMed Central

    Atkinson, John P.; Frank, Michael M.

    1974-01-01

    Purified human IgM isoagglutinins were utilized to sensitize 51Cr-labeled erythrocytes so as to produce a known number of complement-fixing sites. These cells were then reinfused into the erythrocyte donor. A minimum of 20 C1-fixing sites/erythrocyte were required for decreased survival. As the amount of antibody coating the erythrocytes was increased, a larger percentage was sequestered. With 80 C1-fixing sites, more than 75% of the injected erythrocytes were removed from the circulation within 10 min. In each case, the clearance pattern consisted of rapid hepatic sequestration followed by a gradual return of a portion of the erythrocytes into the circulation where they survived normally. Clearance was shown to be dependent upon activation of the classical complement pathway, since sensitized cells survived normally in hereditary angioedema patients with low levels of C4 and no detectable C2. Exposure of sensitized cells to fresh serum for 15 min led to the deposition of 550-800 C3 molecules/C1-fixing site. Such cells were immune adherence positive, were agglutinated by anti-C3b, formed rosettes with human alveolar macrophages, and were sequestered in vivo, presumably because of the interaction of cell-bound C3b with the C3b receptor on hepatic macrophages. After exposure to heated serum as a source of the C3b inactivator, the cells were immune adherence negative, were agglutinated only by anti-C3d, did not form rosettes with macrophages, and survived normally in vivo despite, being Coombs positive. Cleavage of cell-bound C3b to C3d may explain the release phase of the IgM clearance pattern. Whereas erythrocytes coated with IgM antibody and complement were previously thought to be sequestered in the liver because of extensive membrane damage, these experiments suggest that clearance is determined by the interaction of erythrocyte-bound complement fragments with specific receptors on hepatic macrophages. Images PMID:4847248

  11. 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.

  12. 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

  13. Antibodies against a Plasmodium falciparum antigen PfMSPDBL1 inhibit merozoite invasion into human erythrocytes.

    PubMed

    Sakamoto, Hirokazu; Takeo, Satoru; Maier, Alexander G; Sattabongkot, Jetsumon; Cowman, Alan F; Tsuboi, Takafumi

    2012-03-02

    One approach to develop a malaria blood-stage vaccine is to target proteins that play critical roles in the erythrocyte invasion of merozoites. The merozoite surface proteins (MSPs) and the erythrocyte-binding antigens (EBAs) are considered promising vaccine candidates, for they are known to play important roles in erythrocyte invasion and are exposed to host immune system. Here we focused on a Plasmodium falciparum antigen, PfMSPDBL1 (encoded by PF10_0348 gene) that is a member of the MSP3 family and has both Duffy binding-like (DBL) domain and secreted polymorphic antigen associated with merozoites (SPAM) domain. Therefore, we aimed to characterize PfMSPDBL1 as a vaccine candidate. Recombinant full-length protein (rFL) of PfMSPDBL1 was synthesized by a wheat germ cell-free system, and rabbit antiserum was raised against rFL. We show that rabbit anti-PfMSPDBL1 antibodies inhibited erythrocyte invasion of wild type parasites in vitro in a dose dependent manner, and the specificity of inhibitory activity was confirmed using PfMSPDBL1 knockout parasites. Pre-incubation of the anti-PfMSPDBL1 antibodies with the recombinant SPAM domain had no effect on the inhibitory activity suggesting that antibodies to this region were not involved. In addition, antibodies to rFL were elicited by P. falciparum infection in malaria endemic area, suggesting the PfMSLDBL1 is immunogenic to humans. Our results suggest that PfMSPDBL1 is a novel blood-stage malaria vaccine candidate.

  14. Comparison of latex agglutination, wet preparation, and culture for the detection of Trichomonas vaginalis

    PubMed Central

    Adu-Sarkodie, Y; Opoku, B; Danso, K; Weiss, H; Mabey, D

    2004-01-01

    Objectives: To compare the performance of three diagnostic methods for Trichomonas vaginalis infection—latex agglutination, saline wet mount, and culture. Methods: Vaginal swabs from 3807 women attending antenatal clinics were tested for the presence of T vaginalis by latex agglutination. All positives and the following two negatives were tested by wet preparation and culture. Results: The prevalence of infection by latex agglutination was 5.4%. Using an expanded gold standard based on the wet mount and culture results, the sensitivity of the latex agglutination test was 98.8% (95% CI 95.9 to 99.9) and specificity was 92.1 (89.2 to 94.5). The kappa index for test agreement was 0.93 for latex and culture and 0.88 for latex and wet preparation. Conclusion: The latex agglutination test is a highly sensitive test for detecting T vaginalis infection. It is a simple rapid test and has the potential for use in screening and diagnostic settings. PMID:15170003

  15. Hepatitis E Virus in Farmed Rabbits, Wild Rabbits and Petting Farm Rabbits in the Netherlands.

    PubMed

    Burt, Sara A; Veltman, Jorg; Hakze-van der Honing, Renate; Schmitt, Heike; van der Poel, Wim H M

    2016-09-01

    Rabbits have been suggested as a zoonotic source of Hepatitis E virus. Phylogenetic analysis of HEV isolates from farmed, wild and pet rabbits in the Netherlands (23, 0, and 60 % respectively) showed them to be grouped amongst published rabbit HEV sequences and distinct from most human isolates. Dutch rabbits are unlikely to be a zoonotic source.

  16. SERUM SICKNESS IN RABBITS

    PubMed Central

    Fleisher, Mover S.; Jones, Lloyd

    1931-01-01

    1. The injection of a single large dose of normal horse serum into rabbits results in the appearance 3 to 8 days later of erythematous and edematous reactions on the ears in 68.9 per cent of the animals. 2. The injections may be given by any of several routes and reactions appear when the site of injection is definitely distant from the ears. 3. Injections of various antisera into rabbits cause the appearance of similar reactions. 4. These reactions can be considered as manifestations of serum sickness in rabbits. PMID:19869943

  17. Rabbit orthopedic surgery.

    PubMed

    Rich, Gregory A

    2002-01-01

    Orthopedic surgery in rabbits poses several unique parameters for the veterinary surgeon. It is imperative for the veterinarian to be knowledgeable about the anatomic features of the surgical repair site and to become familiar with a rabbit's pain and discomfort often associated with orthopedic injuries. Handling the perioperative and postoperative pain and potential GI disturbances are crucial for a successful outcome of the surgical case. This article is designed to help the veterinary surgeon prepare for the orthopedic surgical procedure and the peripheral physiologic needs of the rabbit from presentation through recovery.

  18. Host antigens on avian oncoviruses: evidence for virus envelope antigens related to specific chicken erythrocyte membrane antigens.

    PubMed

    Aupoix, M; Vigier, P; Blanchet, J P

    1980-01-01

    Avian sarcoma viruses (ASV) of subgroups A to D, produced by chick embryo fibroblasts (CEF), are inactivated to a high degree by rabbit antisera to the membrane antigens of adult chicken and chick embryo erythrocytes, notably by antisera to an antigen of embryo erythrocytes, which is lost by adult erythrocytes and to another antigen specific to the latter erythrocytes. Contrary to virus inactivation by anti-CEF serum reported earlier, virus inactivation by the antisera to these two age-specific antigens does not require complement and is not paralleled by virolysis but by aggregation of virions. The two antigens related, or identical, to the age-specific erythrocyte membrane antigens thus shown to be present on the virus envelope do not pre-exist, or pre-exist only in a low amount, on the CEF membrane, since the virus-inactivating capacity of their antisera is not removed by absorption with CEF. Their appearance on the virus does not depend on cell transformation but only on infection, since both antigens are found on a ts ASV mutant produced at restrictive temperature by untransformed CEF and the virus-inactivating capacity of their antisera is removed by absorption with CEF infected with Rous-associated virus (RAV-1). These findings suggest that infection of CEF by avian oncoviruses may elicit the appearance, or enhance the expression at the cell surface of antigens characteristic of another cell type which may contribute to the formation of specific virus budding sites.

  19. Allometric dependence of the life span of mammal erythrocytes on thermal stability and sphingomyelin content of plasma membranes.

    PubMed

    Ivanov, Ivan Tanev

    2007-08-01

    Thermal stability of erythrocyte membrane is a measure for its ability to maintain permeability barrier at deleterious conditions. Hence, it could impact the resistance of erythrocytes against detrimental factors in circulation. In this study the thermostability of erythrocyte membranes was expressed by the temperature, T(go), at which the transmembrane gradient of ion concentration rapidly dissipated during transient heating. T(go) is the inducing temperature of the membrane transition that activated passive ion permeability at hyperthermia causing thermal hemolysis. A good allometric correlation of T(go) to the resistance against thermal hemolysis and the life span of erythrocytes were found for 13 mammals; sheep, cow, goat, dog, horse, man, rabbit, pig, cat, hamster, guinea pig, rat, and mouse. For the same group, the values of T(go) were strictly related to the sphingomyelin content of erythrocyte membranes. The residual ion permeability, P, was temperature activated from 38 to 57 degrees C with activation energy of 250+/-15 kJ/mol that strongly differed from that below 37 degrees C. The projected value of P at 37 degrees C was about half that of residual physiological permeability for Na+ and K+ that build ground for possible explanation of the life span vs membrane thermostability allometric correlation.

  20. Evaluation of the usefulness of six commercial agglutination assays for serologic diagnosis of toxoplasmosis.

    PubMed

    Villard, Odile; Cimon, Bernard; Franck, Jacqueline; Fricker-Hidalgo, Hélène; Godineau, Nadine; Houze, Sandrine; Paris, Luc; Pelloux, Hervé; Villena, Isabelle; Candolfi, Ermanno

    2012-07-01

    Six agglutination tests for detecting Toxoplasma gondii-specific antibodies (immunoglobulin G or M) in serum were performed and compared. In total, 599 sera were examined using direct and indirect agglutination assays. Sensitivity varied from 93.7% to 100% and specificity from 97.1% to 99.2%. In a selected population with interfering diseases, the percentage of false positives ranged from 4.3% to 10.9%. Although an overall agreement of 100% was found for chronic toxoplasmosis, sensitivity for the detection of confirmed acute toxoplasmosis ranged from 86.4% to 97.3%. Regarding the large variability in terms of the performance of the 6 assays, tests based on the hemagglutination principle were found to be better than the other agglutination tests for all the panels evaluated, meaning that they could be used as qualitative or semiquantitative low-cost screening assays.

  1. Evaluation of the one-point microcapsule agglutination test for diagnosis of leptospirosis.

    PubMed Central

    Arimitsu, Y.; Kmety, E.; Ananyina, Y.; Baranton, G.; Ferguson, I. R.; Smythe, L.; Terpstra, W. J.

    1994-01-01

    We have developed a one-point microcapsule agglutination test (MCAT) for the serodiagnosis of leptospirosis. The MCAT kit was evaluated for use in humans by six WHO Collaborating Centres for Reference and Research on Leptospirosis. The laboratories classified their serum samples on the basis of the microscopic agglutination test (MAT) and the following screening tests: enzyme-linked immunosorbent assay (ELISA), macroscopic (slide) agglutination test, or the complement fixation test. The MCAT may in some instances give a positive result earlier in the course of the disease than MAT or the ELISA IgM; on the other hand, it did not detect antibodies against some serovars, for example, those of the Sejroe or Australis serogroup in Slovakia. In contrast, however, the MCAT detected antibodies to serovar hardjo (the same serogroup as Sejroe) in patients from the United Kingdom and the Russian Federation. PMID:8062397

  2. 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.

  3. Enzymatic Modification of Plasma Low Density Lipoproteins in Rabbits: A Potential Treatment for Hypercholesterolemia

    NASA Astrophysics Data System (ADS)

    Labeque, Regine; Mullon, Claudy J. P.; Ferreira, Joao Paulo M.; Lees, Robert S.; Langer, Robert

    1993-04-01

    Phospholipase A_2 (EC 3.1.1.4) hydrolyzes certain phospholipids of low density lipoprotein (LDL). Plasma clearance of phospholipase A_2-modified human LDL is up to 17 times faster than that of native human LDL in hypercholesterolemic rabbits. Modification of blood lipoproteins of hypercholesterolemic rabbits was performed by using an extracorporeal circuit containing immobilized phospholipase A_2. After 90-min treatments, nearly 30% decreases in plasma cholesterol concentrations were observed. Erythrocyte, leukocyte, and platelet counts showed no net change after treatment. This technique does not require any fluid replacement or sorbent regeneration and offers a potential approach for lowering serum cholesterol and LDL levels.

  4. Agglutinates as recorders of fossil soil compositions. [of Apollo 17 lunar probes

    NASA Technical Reports Server (NTRS)

    Taylor, G. J.; Wentworth, S.; Warner, R. D.; Keil, K.

    1978-01-01

    The composition of agglutinates in polished sections of the Apollo 17 drill core was studied in an attempt to deduce the nature of the Taurus-Littrow valley regolith prior to the formation of the Camelot and Central Cluster craters. The agglutinate compositions in the soils differed from the host soil compositions except for samples from the North Massif. Local materials from the valley floor and the massifs appear to form the pre-Central Cluster regolith. It is also shown that chemical mixing models for bulk soil compositions can be misleading unless the petrologic characteristics of each soil are taken into account.

  5. Human African trypanosomiasis: a latex agglutination field test for quantifying IgM in cerebrospinal fluid.

    PubMed Central

    Lejon, V.; Büscher, P.; Sema, N. H.; Magnus, E.; Van Meirvenne, N.

    1998-01-01

    LATEX/IgM, a rapid agglutination test for the semi-quantitative detection of IgM in cerebrospinal fluid of patients with African trypanosomiasis, is described in this article. The lyophilized reagent has been designed for field use and remains stable at 45 degrees C for one year. The test has been evaluated on cerebrospinal fluid samples from trypanosome-infected and non-infected patients, by comparison with commercial latex agglutination, radial immunodiffusion, and nephelometry. All test systems yielded similar results. PMID:10191550

  6. [Relationship between the sensitivity of the delayed agglutination test and synthetic detergents].

    PubMed

    Iovchev, E; Vodas, K

    1977-01-01

    Residual amounts of detergents (Losk, Bio-73, Alka-lux, Bourgas, Bourgaslux, and Vero) in a concentration of 10-5 to 10-7 in physiologic saline can inhibit the agglutination titers by 3 to 5 degrees. This could mislead in the assessment of the reaction with regard to its diagnostic value. It is admitted that the inhibition produced is due to changes in the antibodies--drop in the total protein and light variations in all protein fractions as well as in the probable surface deterioration of the antigen, leading to its defective agglutinability. It is suggested to rinse more than five times all glassware that has been cleaned with detergents.

  7. [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.

  8. Prevalence of naturally occurring antibodies against dog erythrocyte antigen 7 in a population of dog erythrocyte antigen 7-negative dogs from Spain and Italy.

    PubMed

    Spada, Eva; Proverbio, Daniela; Viñals Flórez, Luis Miguel; Del Rosario Perlado Chamizo, Maria; Serra Y Gómez de la Serna, Blanca; Perego, Roberta; Baggiani, Luciana

    2016-08-01

    OBJECTIVE To determine the prevalence of naturally occurring anti-dog erythrocyte antigen (DEA) 7 antibodies in DEA 7-negative dogs from Spain and Italy. ANIMALS 252 DEA 7-negative dogs from a population of 312 dogs that were previously tested for DEA 1, DEA 4, and DEA 7. PROCEDURES A plasma sample was obtained from each dog and evaluated for anti-DEA 7 antibodies by the use of gel column agglutination. Each plasma sample underwent major crossmatching with RBCs from DEA 7-positive dogs. Samples that resulted in agglutination were then crossmatched with RBCs from DEA 1-negative, DEA 4-positive, and DEA 7-negative dogs to confirm the presence of anti-DEA 7 antibodies. Results were then used to calculate the risk for a delayed transfusion reaction in a DEA 7-negative dog with anti-DEA 7 antibodies after a transfusion with blood that was not crossmatched or typed for DEA 7. RESULTS 96 of 252 (38.1%) plasma samples contained anti-DEA 7 antibodies. A DEA 7-negative dog with anti-DEA 7 antibodies had a 5.9% chance of developing a delayed hemolytic reaction after transfusion with blood not crossmatched or typed for DEA 7. CONCLUSIONS AND CLINICAL RELEVANCE Results indicated that canine blood used for transfusion should be crossmatched with the blood or plasma of the intended recipient prior to transfusion to minimize the likelihood that the recipient will develop a hemolytic reaction associated with anti-DEA 7 antibodies. Ideal canine blood donors should be negative for both DEA 1 and DEA 7.

  9. The changes in serum and whole blood rheological properties of rabbits during the progression of atherosclerosis.

    PubMed

    K Abdelhalim, Mohamed Anwar; Al-Ayed, Mohammed Suliman; Abdelmottaleb Moussa, Sherif A; Al-Mohy, Yanallah Hussain

    2016-05-01

    This study aimed to evaluate the role of zinc (Zn)-supplemented with high cholesterol diet (HCD) on the serum and whole blood rheological properties of rabbits fed a HCD. Twenty-four New Zealand white rabbits were divided into three groups. The HCD group was fed a diet with 1.0% cholesterol and 1.0% olive oil. The HCD + Zn group was fed a diet with 1.0% cholesterol, 1.0% olive oil, and Zn. Blood viscosity, shear stress, and torque (%) were measured at shear rates ranging from 225 to 1875 s-1 for serum and 75-900 s-1 for whole blood. Serum viscosity and shear stress in HCD rabbits were significantly higher at all shear rates compared to controls; while whole blood viscosity and shear stress in HCD rabbits were significantly lower at all shear rates compared to controls. Viscosity and shear stress in both serum and whole blood from rabbits in the HCD + Zn group returned to normal values at all shear rates. The Zn supplemented to HCD rabbits, delays the progression of atherosclerosis. Changes in blood serum viscosity could reflect changes in non-clotting proteins, glucose, nutrients and trace elements; while changes in whole blood viscosity could result from changes in hematocrit, hemoglobin, and erythrocyte count. One of the factors responsible for increasing the serum viscosity values of HCD rabbits might be attributed to increase in Fe and decrease in Zn levels in the blood serum.

  10. Real time observation and automated measurement of red blood cells agglutination inside a passive microfluidic biochip containing embedded reagents.

    PubMed

    Huet, Maxime; Cubizolles, Myriam; Buhot, Arnaud

    2016-09-20

    The process of agglutination is commonly used for the detection of biomarkers like proteins or viruses. The multiple bindings between micrometer sized particles, either latex beads or red blood cells (RBCs), create aggregates that are easily detectable and give qualitative information about the presence of the biomarkers. In most cases, the detection is made by simple naked-eye observation of agglutinates without any access to the kinetics of agglutination. In this study, we address the development of a real-time time observation of RBCs agglutination. Using ABO blood typing as a proof-of-concept, we developed i) an integrated biological protocol suitable for further use as point-of-care (POC) analysis and ii) two dedicated image processing algorithms for the real-time and quantitative measurement of agglutination. Anti-A or anti-B typing reagents were dried inside the microchannel of a passive microfluidic chip designed to enhance capillary flow. A blood drop deposit at the tip of the biochip established a simple biological protocol. In situ agglutination of autologous RBCs was achieved by means of embedded reagents and real time agglutination process was monitored by video recording. Using a training set of 24 experiments, two real-time indicators based on correlation and variance of gray levels were optimized and then further confirmed on a validation set. 100% correct discrimination between positive and negative agglutinations was performed within less than 2min by measuring real-time evolution of both correlation and variance indicators.

  11. 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.

  12. 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.

  13. Extracellular calcium is involved in egg yolk-induced head-to-head agglutination of bull sperm.

    PubMed

    Yang, D H; McMillan, A G; Standley, N T; Shannon, P; Xu, Z Z

    2012-10-15

    Head-to-head agglutination of bull sperm occurs when semen is highly diluted in an egg yolk-citrate diluent without streptomycin. The objectives were to investigate causes of sperm agglutination and the underlying mechanism. Aliquots of bull semen were diluted in a base diluent (BD) supplemented with various test components and the percentage of agglutinated sperm (% AggSp) was quantified at 1, 5, 24, 48, and 72 h of incubation. When sperm were incubated at 22 °C, no agglutination was observed in BD for up to 72 h, whereas the % AggSp was 5.0, 41.7, 72.2, 91.1, and 92.8% in BD + 5% egg yolk (BD + EY) at 1, 5, 24, 48 and 72 h, respectively. However, no sperm agglutination was observed in BD + EY if incubation temperature was 37 °C. Addition of 5 or 10 mM ethylenebis (oxyethyleneni-trilo) tetra-acetic acid to BD + EY reduced the % AggSp from 95% to <5% at 72 h (P < 0.001), but addition of 5 mM CaCl(2) to BD failed to induce sperm agglutination in the absence of egg yolk, implicating calcium and other factors in egg yolk. Addition of the citrate-soluble fraction (CSF) of egg yolk to BD induced sperm agglutination similar to whole egg yolk, whereas water- and saline-soluble fractions of egg yolk were ineffective. The sperm-agglutinating efficacy of CSF (the % AggSp = 95% at 72 h) was reduced by dialysis (20%; P < 0.05), partially restored by addition of 5 mM CaCl2 (70%; P < 0.05), but the calcium effect was neutralized by addition of 5 mM ethylenebis (oxyethyleneni-trilo) tetra-acetic acid (1.7%; P < 0.05), again implicating calcium. Addition of 30 μM of a protein kinase A inhibitor (H-89) to an agglutinating diluent failed to inhibit sperm agglutination, whereas addition of 2 mM of a cAMP analogue, dbcAMP, to a nonagglutinating diluent failed to induce sperm agglutination. Agglutination status had no effect on sperm plasma membrane/acrosome status and mitochondrial membrane potential. In conclusion, calcium and other component(s) in the CSF of egg yolk induced head

  14. 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

  15. Single agglutinates: A comparative study of compositions of agglutinitic glass, whole-grain, bulk soil, and FMR

    NASA Technical Reports Server (NTRS)

    Basu, A.; Robinson, R.; Mckay, D. S.; Blanchard, D. P.; Morris, R. V.; Wentworth, Susan J.

    1994-01-01

    Previous workers on single agglutinates have variously interpreted the composition of agglutinitic glass to represent impact melts of (1) bulk soil, (2) mixed components in finer sizes, and (3) microtargets. Separately, Papike has argued in favor of fusion of the finest fraction of bulk soils. Thirty-four single agglutinates were hand-picked from the mature Apollo 16 soil 61181 (I(sub s)/FeO = 82) and the FMR and chemical composition (INAA for Fe, Sc, Sm, Co, Ni, and Cr) of each agglutinate particle were measured. Thirteen of these single agglutinates were selected for electron beam microanalysis and imaging. Less than 1 micron spots were analyzed (for Na, Mg, Al, Si, P, S, K, Ca, Ti, Cr, Mn, Fe, Ni, and Ba) on pure glassy areas (approximately ten in each particle) selected on the basis of optical and BSE images (avoiding all clasts and inclusions) with an electron microprobe to obtain average glass compositions of each single agglutinate.

  16. 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.

  17. [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.

  18. Characterization of two different agglutinators in the latex fixation test, occurring in normal human sera

    PubMed Central

    Klein, F.; Valkenburg, H. A.; Van Zwet, Theda L.; Lafeber, Geertruida J. M.

    1966-01-01

    Using a sensitive modification of the latex fixation test it is possible to detect a small agglutinating effect in about 60 per cent of normal human sera, after these have been heated for 30 minutes at 56°. This was shown to be caused by an IgM globulin with the properties of a rheumatoid factor. The factor is able to react with human IgG globulin and may represent an antibody to the IgG part of circulating antigen—antibody complexes. The heat treatment probably inactivates an inhibitor of the latex fixation reaction. In addition all normal human sera give an agglutination reaction with IgG coated latex at incubation temperatures of 37° or lower. It was shown that these reactions are caused by a thermolabile, non-reducible component with a sedimentation constant of about 10. This component is probably identical with the complement component C'1q. The agglutinating activity was found in the α2—β1 region after electrophoresis of untreated serum, but in the slow γ region after treatment of the serum with EDTA. This kind of agglutination may cause false positive reactions in latex tests which are carried out at 37° or less. ImagesFIG. 1FIG. 3 PMID:4160336

  19. Cross-reactivity in Cryptococcus antigen latex agglutination test in two commercial kits.

    PubMed

    Tone, Kazuya; Umeda, Yoshiko; Makimura, Koichi

    2016-05-01

    This article presents an examination of the cross-reactivity of pathogenic fungi with Cryptococcus neoformans in two commercial Cryptococcus antigen latex agglutination tests performed across 39 fungal strains. Some fungi were newly indicated as Cryptococcus cross-reactive, and the two kits showed differences in cross-reactive fungi.

  20. The Classroom-Friendly ABO Blood Types Kit: Blood Agglutination Simulation

    ERIC Educational Resources Information Center

    Arnold, Savittree Rochanasmita; Kruatong, Tussatrin; Dahsah, Chanyah; Suwanjinda, Duongdearn

    2012-01-01

    The classroom-friendly ABO blood type kit was developed by combining advantages of modelling and a simulation laboratory to teach the topics of ABO blood types and blood transfusion. Teachers can easily simulate the agglutination reaction on a blood type testing plate in the classroom, and show the students how this reaction occurs by using the…

  1. Reading Development in Agglutinative Languages: Evidence from Beginning, Intermediate, and Adult Basque Readers

    ERIC Educational Resources Information Center

    Acha, Joana; Laka, Itziar; Perea, Manuel

    2010-01-01

    Do typological properties of language, such as agglutination (i.e., the morphological process of adding affixes to the lexeme of a word), have an impact on the development of visual word recognition? To answer this question, we carried out an experiment in which beginning, intermediate, and adult Basque readers (n = 32 each, average age = 7, 11,…

  2. 9 CFR 147.1 - The standard tube agglutination test. 1

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ..., high agglutinability, but are not sensitive to negative and nonspecific sera. The stock cultures may be... hours at 37 °C. The antigenic composition and purity of the stock cultures should be checked... cultures prepared from the stock cultures of the selected strains. The antigen-growing tubes or...

  3. 9 CFR 147.1 - The standard tube agglutination test. 1

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ..., high agglutinability, but are not sensitive to negative and nonspecific sera. The stock cultures may be... hours at 37 °C. The antigenic composition and purity of the stock cultures should be checked... cultures prepared from the stock cultures of the selected strains. The antigen-growing tubes or...

  4. Monoclonal antibodies for the detection of desialylation of erythrocyte membranes during haemolytic disease and haemolytic uraemic syndrome caused by the in vivo action of microbial neuraminidase.

    PubMed

    Seitz, R C; Poschmann, A; Hellwege, H H

    1997-09-01

    Especially in childhood, the in vivo action of microbial neuraminidase may cause haemolytic anaemia or life-threatening haemolytic uraemic syndrome. The exposure of the Thomsen-Friedenreich (T) crypto-antigen and T-antigen polyagglutinability of erythrocytes has been described as the first sign of toxic cleavage of N-acetylneuraminic acid (Neu5Ac) from sialoglycoproteins of cell membranes. This phenomenon may, however, be too unspecific to initiate treatment for toxin elimination. The present study investigated the diagnostic effectiveness of a panel of three monoclonal antibodies (mcabs) for the estimation of the clinical significance of neuraminidase action in vivo. Depending on the amount of Neu5Ac released, the mcabs I-C4, II-Q9 and III-Y12 recognized different epitopes on erythrocyte asialoglycophorin. In 1345 patients, the mcab II-09 detected cleavage of Neu5Ac in 32 children who had T-antigen polyagglutinability and mild to moderate haemolytic anaemia. However, only 10 patients, whose erythrocytes were agglutinated by the mcabs III-Y12 or I-C4, developed severe haemolysis, thrombocytopenia, and finally the life-threatening haemolytic uraemic syndrome (p<0.0002). In conclusion, these mcabs provided an early marker of the in vivo action of neuraminidase. Two different degrees of erythrocyte desialylation, as defined by these mcabs, are suggested to reflect the severity of toxin-associated disease.

  5. Erythrocytes and microbubble contrast agents, improve the therapeutic efficiency of high intensity focused ultrasound

    NASA Astrophysics Data System (ADS)

    Takegami, Kenji; Kaneko, Yukio; Watanabe, Toshiaki; Maruyama, Toshiyuki; Matsumoto, Yoichiro; Nagawa, Hirokazu

    2005-03-01

    Erythrocytes, an well as Levovist microbubble contrast agent, enhance the heating effect of high intensity focused ultrasound (HIFU) and increase the coagulation volume produced by HIFU irradiation. In vitro experiments used human plasma with various concentrations of human erythrocytes in combination with or without Levovist. In vivo experiments used eight Japan white rabbits with three levels of anaemia. Using a 2.17 MHz transducer, HIFU was applied for 60 seconds, and the temperature rise and the volume of coagulation necrosis was evaluated. There was a significant correlation between the HIFU-induced temperature rise and hematocrit, with a correlation coefficient of 0.998 (p=0.0001). Although the temperature rise was smaller at low hematocrit, it was significantly increased by adding Levovist to the suspension (p<0.01). The mean volume of coagulation necrosis was significantly higher in the rabbits with higher hematocrit (p<0.01), and that in the moderate anaemia group was significantly increased by using Levovist (p<0.01).

  6. The Production of Nominal and Verbal Inflection in an Agglutinative Language: Evidence from Hungarian

    PubMed Central

    Peckham, Don; Szanka, Szilvia; Gazso, Dorottya; Lovassy, Noemi; Ullman, Michael T.

    2015-01-01

    The contrast between regular and irregular inflectional morphology has been useful in investigating the functional and neural architecture of language. However, most studies have examined the regular/irregular distinction in non-agglutinative Indo-European languages (primarily English) with relatively simple morphology. Additionally, the majority of research has focused on verbal rather than nominal inflectional morphology. The present study attempts to address these gaps by introducing both plural and past tense production tasks in Hungarian, an agglutinative non-Indo-European language with complex morphology. Here we report results on these tasks from healthy Hungarian native-speaking adults, in whom we examine regular and irregular nominal and verbal inflection in a within-subjects design. Regular and irregular nouns and verbs were stem on frequency, word length, and phonological structure, and both accuracy and response times were acquired. The results revealed that the regular/irregular contrast yields similar patterns in Hungarian, for both nominal and verbal inflection, as in previous studies of non-agglutinative Indo-European languages: the production of irregular inflected forms was both less accurate and slower than of regular forms, both for plural and past-tense inflection. The results replicate and extend previous findings to an agglutinative language with complex morphology. Together with previous studies, the evidence suggests that the regular/irregular distinction yields a basic behavioral pattern that holds across language families and linguistic typologies. Finally, the study sets the stage for further research examining the neurocognitive substrates of regular and irregular morphology in an agglutinative non-Indo-European language. PMID:25769039

  7. Interactions with lectins and agglutination profiles of clinical, food, and environmental isolates of Listeria.

    PubMed Central

    Facinelli, B; Giovanetti, E; Casolari, C; Varaldo, P E

    1994-01-01

    On the basis of preliminary trials with 14 collection strains of Listeria, five lectins (Canavalia ensiformis, concanavalin A; Griffonia simplicifolia lectin I; Helix pomatia agglutinin; Ricinus communis agglutinin; and Triticum vulgaris wheat germ agglutinin) were selected to set up a microtiter agglutination assay. The lectin agglutination profiles of 174 clinical, food, and environmental strains of Listeria monocytogenes, Listeria innocua, and Listeria seeligeri were investigated. Data on the standard determination of the antigenic structure were available for clinical strains; nonclinical isolates were assigned to serogroup 1 or 4 with commercial antisera. The listeria-lectin interaction was related to serological type rather than species; in particular, the strains assigned to serogroup 1 or belonging to serovars 1/2a, 1/2b, 1/2c, 3a, 3b, and 7 were never agglutinated by G. simplicifolia lectin I. The five-lectin set proved to be capable of detecting differences between serologically identical isolates of L. monocytogenes. Of the 150 isolates of this species, 144 were distributed over 15 different lectin agglutination profiles and 6 autoagglutinated, the overall typeability being 96%. However, the profiles encountered among L. monocytogenes isolates were not randomly distributed. With strains assigned to serogroup 1 or belonging to serovars 1/2a, 1/2b, 1/2c, and 3b, the clinical isolates fell into only two of the eight patterns recorded overall; with strains of serogroup 4 and serovar 4b, food and environmental isolates were distributed over eight of the nine patterns found in total, while clinical isolates were distributed over five patterns. In a comparative study of 15 epidemiologically relevant isolates of L. monocytogenes from five distinct outbreaks, strains with identical phage types and/or DNA fingerprints displayed identical lectin profiles. The heterogeneity of agglutination profiles may form the basis of a new approach to L. monocytogenes typing

  8. The production of nominal and verbal inflection in an agglutinative language: evidence from Hungarian.

    PubMed

    Nemeth, Dezso; Janacsek, Karolina; Turi, Zsolt; Lukacs, Agnes; Peckham, Don; Szanka, Szilvia; Gazso, Dorottya; Lovassy, Noemi; Ullman, Michael T

    2015-01-01

    The contrast between regular and irregular inflectional morphology has been useful in investigating the functional and neural architecture of language. However, most studies have examined the regular/irregular distinction in non-agglutinative Indo-European languages (primarily English) with relatively simple morphology. Additionally, the majority of research has focused on verbal rather than nominal inflectional morphology. The present study attempts to address these gaps by introducing both plural and past tense production tasks in Hungarian, an agglutinative non-Indo-European language with complex morphology. Here we report results on these tasks from healthy Hungarian native-speaking adults, in whom we examine regular and irregular nominal and verbal inflection in a within-subjects design. Regular and irregular nouns and verbs were stem on frequency, word length, and phonological structure, and both accuracy and response times were acquired. The results revealed that the regular/irregular contrast yields similar patterns in Hungarian, for both nominal and verbal inflection, as in previous studies of non-agglutinative Indo-European languages: the production of irregular inflected forms was both less accurate and slower than of regular forms, both for plural and past-tense inflection. The results replicate and extend previous findings to an agglutinative language with complex morphology. Together with previous studies, the evidence suggests that the regular/irregular distinction yields a basic behavioral pattern that holds across language families and linguistic typologies. Finally, the study sets the stage for further research examining the neurocognitive substrates of regular and irregular morphology in an agglutinative non-Indo-European language.

  9. Commercial latex agglutination test for rapid diagnosis of group B streptococcal infection in infants.

    PubMed Central

    Webb, B J; Baker, C J

    1980-01-01

    Although latex agglutination assays for detection of a variety of bacterial antigens in body fluids from patients with systemic infection have been shown to be useful as rapid diagnostic techniques, lack of commercial availability has restricted their application. The Streptex latex test kit for the detection of group B streptococcal (GBS) antigen in admission body fluid specimens was evaluated for sensitivity and specificity in 54 infants with meningitis and in 10 infants with normal cerebrospinal fluid (CSF) parameters. GBS antigen was detected in 22 of 28 (78.6%) CSF specimens by latex agglutination and in 23 of 28 (82.1%) by countercurrent immunoelectrophoresis. Antigen was present in 21 of 28 (latex agglutination) and 19 of 26 (countercurrent immunoelectrophoresis) CSF specimens after the initiation of antimicrobial therapy. Heat-labile factors accounted for nonspecific agglutination reactions with latex suspensions other than group B in 3 of 28 CSF samples from patients with GBS meningitis. These nonspecific reactions were readily eliminated by heating specimens for 10 min at 100 degrees C. Fifteen patients with GBS meningitis had admission serum and urine samples collected in addition to CSF. Antigen was detected by latex agglutination and countercurrent immunoelelectrophoresis in 14 of 15 (93.3%) and 13 of 15 (86.7%) concentrated urine specimens, respectively, and in 12 of 15 (80%) CSF specimens and 4 of 15 (27%) sera by each method. These findings indicate that the Streptex latex test is a rapid, sensitive, and readily available method for detection of GBS antigen in admission body fluid specimens from infants with meningitis. PMID:7012177

  10. Induction of tolerance by haptenated liposomes carrying mouse erythrocyte membrane glycoprotein.

    PubMed Central

    Hitsumoto, Y; Hineno, T; Utsumi, S

    1984-01-01

    Membrane sialoglycoprotein (GP) of mouse erythrocytes were incorporated into artificial membranes of liposomes which also contained DNP-aminocaproylphosphatidylethanolamine hapten. The liposomes were tested for their potential to modulate the anti-hapten antibody response in mice as a plausible model of haptenated isologous erythrocytes which is known to be a potent tolerogen. Repeated intravenous inoculation with DNP-liposomes carrying mouse GP resulted in a significant suppression of splenic plaque-forming cells and serum antibody produced in the recipient mice against the subsequent immunization with DNP-KLH in saline or DNP-HGG in Freund's complete adjuvant, in a strictly hapten-specific fashion. The suppression was long-lasting, and the secondary response to DNP-KLH given over a month later was also affected. DNP-liposomes without GP, or those with rabbit GP, exhibited little or no suppressive effect. The observed capacity of mouse GP-bearing DNP-liposomes was reminiscent of that of haptenated mouse erythrocytes in that only the IgG response, but not the IgM response, was suppressed. Images Figure 1 PMID:6209210

  11. THE OSMOTICALLY FUNCTIONAL WATER CONTENT OF THE HUMAN ERYTHROCYTE.

    PubMed

    LEFEVRE, P G

    1964-01-01

    Experiments were directed toward estimation of the magnitude of error incurred by the presumption of idealized osmometric behavior in the author's recent studies of monosaccharide transport through the human erythrocyte membrane. Thick suspensions of washed cells in isotonic buffered balanced salt medium were mixed in fixed proportions with varying dilutions of a concentrate of either (a) the mixed chlorides of the medium, or (b) glucose in the isotonic medium, and the resultant freezing point and hematocrit values determined. The form of the responses in the tonicity and the cell volume, as functions of the variable dilution of sugar or salts, conformed consistently with relations derived from the classical van't Hoff-Boyle-Mariotte pressure-volume relation. However, the effective cell water contents appeared substantially less than the weight lost in conventional drying, and varied somewhat according to the index used: expressed as grams of H(2)O per milliliter of cells at isotonic volume, the cell water implied by the hematocrit behavior was 0.614 +/- 0.015 (SD); by the salt tonicity response, 0.565 +/- 0.027; by the immediate glucose tonicity response, 0.562 +/- 0.044; and by the equilibrium glucose tonicities, 0.589 +/- 0.043. Olmstead's reports of gross deviation from the van't Hoff relation, in the rabbit red cell's responses to tonicity displacement, are attributed primarily to a systematic aberration in his method of data analysis, the observations themselves agreeing substantially with the present findings.

  12. Complete genome sequence of two rabbit hemorrhagic disease virus variant b isolates detected on the Iberian Peninsula.

    PubMed

    Dalton, K P; Abrantes, J; Lopes, A M; Nicieza, I; Álvarez, Á L; Esteves, P J; Parra, F

    2015-03-01

    We report the complete genome sequences of two isolates (RHDV-N11 and CBVal16) of variant rabbit hemorrhagic disease virus (RHDVb). Isolate N11 was detected in young domestic animals during a rabbit hemorrhagic disease (RHD) outbreak that occurred in 2011 on a rabbit farm in Navarra, Spain, while CBVal16 was isolated from a wild rabbit found dead in Valpaços, Northern Portugal, a year later. The viral sequences reported show 84.8-85.1 % and 78.3-78.5 % identity to RHDVAst/89 and RCV-A1 MIC-07, representative members of the pathogenic genogroup 1 RHDV and apathogenic rabbit calicivirus, respectively. In comparison with other RHDV isolates belonging to the previously known genogroups 1-6, RHDVb shows marked phenotypic differences, as it causes disease preferentially in young rabbits under 40 days of age and shows modified red blood cell agglutination profiles as well as antigenic differences that allow this variant to escape protection by the currently available vaccines.

  13. [Evaluation of the usefulness of the agglutination test with Mangifera indica extract for the identification of pathogenic Yersinia enterocolitica strains].

    PubMed

    Kałuzewski, S; Gierczyński, R; Szych, J; Jagielski, M

    1997-01-01

    The study was performed on 137 Y. enterocolitica strains belonging to various serological groups, including 75 03 group strains isolated form human clinical material. The agglutination test on slides was carried out on this strains using Mangifera indica extract of own production. Agglutinating preparation obtained from the seeds of M. indica agglutinated Y. enterocolitica organisms possessing the pVY plasmid and CRMOX+ phenotype in dilutions to 1.56 micrograms/ml. In identification tests conducted parallelly agglutination solution was used in concentrations of 100 and 10 micrograms/ml. All clones of Y. enterocolitica from O3 group from cultures at 37 degrees C and with CRMOX+ phenotype possessing the pVY plasmid were agglutinated by the extract. Agglutination failed to develop in the cultures of these clones incubated at 25 degrees C. Yersinia clones not containing the pVY plasmid with CRMOX- phenotype were resistant to agglutination. The virulence plasmid was found in 44 out of 75 strains of Y. enterocolitica O3 and was identified by restriction analysis after plasmid DNA digestion with Eco RI enzyme. The obtained results agreed with those of Wauters et al. in 1995 and confirmed the opinion of these authors on the usefulness of the test with M. indica agglutinin for the identification of virulent Y. enterocolitica strains.

  14. Comparison of gel column, card, and cartridge techniques for dog erythrocyte antigen 1.1 blood typing

    PubMed Central

    Seth, Mayank; Jackson, Karen V.; Winzelberg, Sarah; Giger, Urs

    2012-01-01

    Objective To compare accuracy and ease of use of a card agglutination assay, an immunochromatographic cartridge method, and a gel-based method for canine blood typing. Sample Blood samples from 52 healthy blood donor dogs, 10 dogs with immune-mediated hemolytic anemia (IMHA), and 29 dogs with other diseases. Procedures Blood samples were tested in accordance with manufacturer guidelines. Samples with low PCVs were created by the addition of autologous plasma to separately assess the effects of anemia on test results. Results Compared with a composite reference standard of agreement between 2 methods, the gel-based method was found to be 100% accurate. The card agglutination assay was 89% to 91% accurate, depending on test interpretation, and the immunochromatographic cartridge method was 93% accurate but 100% specific. Errors were observed more frequently in samples from diseased dogs, particularly those with IMHA. In the presence of persistent autoagglutination, dog erythrocyte antigen (DEA) 1.1 typing was not possible, except with the immunochromatographic cartridge method. Conclusions and Clinical Relevance The card agglutination assay and immunochromatographic cartridge method, performed by trained personnel, were suitable for in-clinic emergency DEA 1.1 blood typing. There may be errors, particularly for samples from dogs with IMHA, and the immunochromatographic cartridge method may have an advantage of allowing typing of samples with persistent autoagglutination. The laboratory gel-based method would be preferred for routine DEA 1.1 typing of donors and patients if it is available and time permits. Current DEA 1.1 typing techniques appear to be appropriately standardized and easy to use. PMID:22280380

  15. Clinical evaluation of the QuickVet/RapidVet canine dog erythrocyte antigen 1.1 blood-typing test.

    PubMed

    Kohn, Barbara; Classe, Gabriele; Weingart, Christiane

    2012-05-01

    In transfusion medicine, blood typing is an integral part of pretransfusion testing. The objective of the current study was the clinical evaluation of an automated canine cartridge dog erythrocyte antigen (DEA) 1.1 blood-typing method (QuickVet/RapidVet) and comparison of the results with a gel column-based method (ID-Gel Test Canine DEA 1.1). Ethylenediamine tetra-acetic acid-anticoagulated blood samples from 11 healthy and 85 sick dogs were available for typing. Before blood typing, all samples were tested for agglutination and hemolysis. All samples were tested once or multiple times with both methods according to the manufacturer's guidelines. With the gel method, 53 dogs tested DEA 1.1 positive and 42 dogs DEA 1.1 negative; blood typing was not possible due to erythrocyte autoagglutination in 1 dog. With the cartridge test, 53 samples tested DEA 1.1 positive, 34 samples tested DEA 1.1 negative, and 6 results were inconclusive (3 samples were not included due to autoagglutination or severe hemolysis). Without taking the inconclusive samples into account, the agreement between both methods was 96.5%. The sensitivity and specificity for samples that were definitively typed by both methods were 100% and 91.9%, respectively. The cartridge test was suitable for in-clinic canine DEA 1.1 blood typing, although some discrepancies compared to the gel method existed. The cartridge test is software-directed, is easy to use, and does not require user interpretation, but preanalytical guidelines (sample evaluation for agglutination and hemolysis) have to be followed. For inconclusive results, an alternate blood-typing method should be performed.

  16. Drug-induced erythrocyte membrane internalization

    PubMed Central

    Ben-Bassat, Isaac; Bensch, Klaus G.; Schrier, Stanley L.

    1972-01-01

    In vitro erythrocyte membrane internalization, resulting in the formation of membrane-lined vacuoles, can be quantified by a radioisotopic method. A complex of 37Co-labeled vitamin B12 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. Images PMID:4555785

  17. 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.

  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. 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.

  20. Prevalence of dog erythrocyte antigens 1, 4, and 7 in galgos (Spanish Greyhounds).

    PubMed

    Spada, Eva; Proverbio, Daniela; Viñals Flórez, Luis Miguel; Del Rosario Perlado Chamizo, Maria; Perego, Roberta; Bagnagatti De Giorgi, Giada; Baggiani, Luciana

    2015-07-01

    Galgos (Spanish Greyhounds), in common with other sighthounds, have higher hematocrits, hemoglobin concentrations, and red blood cell counts than other breeds. In addition to these hematological characteristics, the physical characteristics of these dogs (medium to large dogs with an easily accessible jugular vein and a good temperament) make galgos ideal blood donors. However, to date, there are only published reports concerning dog erythrocyte antigen (DEA) 1 in this breed. Information on DEAs 4 and 7 would be useful when recruiting blood donors to donation programs, as DEA 1 and 7-negative and DEA 4-positive dogs can be considered universal donors. Ethylenediamine tetra-acetic acid-anticoagulated jugular blood samples were collected from 205 galgos. Dogs were aged between 1 and 10 years, 102 were female (49.8%) and 103 male (50.2%), and all were living in South Madrid, Spain. All 205 blood samples were tested for DEA 1 by card agglutination, and 150 of these samples were tested for DEA 4 and DEA 7 by gel column agglutination using polyclonal anti-DEA antibodies. Of the 205 galgos blood samples typed, 112 out of 205 (54.6%) were positive for DEA 1. Of the 150 blood samples tested, all (150/150, 100%) were positive for DEA 4, and 12 out of 150 (8%) samples tested positive for DEA 7. Of these samples, 70 out of 150 (46.7%) were positive only for DEA 4. There was no relationship between blood types and sex. In addition to the hematological characteristics previously reported and the physical characteristics of these dogs, the relative prevalence of blood types DEA 1, 4, and 7 make galgos good candidates for blood donation in blood donor programs.

  1. [Diagnosis of some yeasts in Metschnikowia genus with the aid of Salmonella cholerae-suis O agglutinating serum (author's transl)].

    PubMed

    Aksoycan, N

    1980-04-01

    In this paper a common antigenic factor among Salmonella choleraesuis 0 antigen and standard Metschnikowia bicuspidata var. bicuspidata and M. pulcherrima strains is shown. This common factor was not present in M. bicuspidata var. australis, M. bicuspidata var. california, M. krissii and M. reukaufii strains. M. bicuspidata var. chathamia and M. zobellii showed agglutination in the previous experiments. According to these results, the use of S. choleraesuis 0:6,7 agglutinating serum for slide and tube agglutination tests can be a diagnostic aid for typing above mentioned Metschnikowia strains along with the other tests.

  2. Elemental X-ray mapping of agglutinated foraminifer tests: a non- destructive technique for determining compositional characteristics.

    USGS Publications Warehouse

    Commeau, R.F.; Reynolds, Leslie A.; Poag, C.W.

    1985-01-01

    The composition of agglutinated foraminiferal tests vary remarkably in response to local substrate characteristics, physiochemical properties of the water column and species- dependant selectivity of test components. We have employed a technique that combines a scanning electron microscope with an energy dispersive X-ray spectrometer system to identify major and minor elemental constituents of agglutinated foraminiferal walls. As a sample is bombarded with a beam of high energy electrons, X-rays are generated that are characteristic of the elements present. As a result, X- ray density maps can be produced for each of several elements present in the tests of agglutinated foraminifers. -Authors

  3. A B-lymphoma cell line that forms rosettes with neuraminidase-treated sheep erythrocytes through monoclonal surface immunoglobulin.

    PubMed

    Tsutsumi, Y; Suzuki, S; Mikata, A; Suzuki, H; Kageyama, K; Watanabe, S; Minato, K; Shimoyama, M

    1982-06-01

    Undifferentiated lymphoma from a 39-year-old female became serially xenotransplantable to preirradiated nude mice. The tumor cells (KT) possessed a monoclonal surface immunoglobulin (SIg mu, kappa) and formed rosettes with neuraminidase-treated sheep erythrocytes (SEn). Precise characterizations of the SEn rosette, however, revealed the following facts: (1) Neuraminidase-untreated or 2-aminoethylisothiuronium bromide (AET) treated sheep erythrocytes were not bound to the KT cells. (2) SEn rosettes on the KT cells did not show a temperature dependency. (3) Neuraminidase-treated erythrocytes from man, horse, mouse, and rabbit were not bound to the KT cells. (4) Preincubation of the KT cells with antipolyvalent immunoglobulin or anti-kappa-chain serum abolished the SEn rosette formation. (5) Trypsinization decreased both SEn rosettes and SIg on the KT cells. (6) SEn rosettes on the KT cells were too loose to be separated from nonrosetting cells by a Percoll gradient centrifugation method. Summarizing these results, the monoclonal SIg on the KT cells recognized sheep erythrocyte antigen(s) that were exposed only after the neuraminidase treatment. Therefore, this was considered to be a case with peculiar B-lymphoma cells that bound SEn through their SIg.

  4. Use of microchip-based hydrodynamic focusing to measure the deformation-induced release of ATP from erythrocytes.

    PubMed

    Moehlenbrock, Michael J; Price, Alexander K; Martin, R Scott

    2006-08-01

    In order to understand the role that erythrocytes play in conditions such as pulmonary hypertension, in vitro mimics of the microcirculation are needed. This paper describes the use of microchip-based hydrodynamic focusing to develop a mimic that allows both mechanical deformation of erythrocytes and quantification of the adenosine triphosphate (ATP) that is subsequently released in response to this deformation. In this mimic, two sheathing streams of a luciferin/luciferase mixture are used to focus and deform a central fluid flow of an erythrocyte sample. The focusing width is changed by simply manipulating the sheath flow rate. This allows a variety of cross-sectional areas to be studied using single point chemiluminescent detection. It was shown that increasing the sheath flow rate does result in elevated levels of ATP release. For example, one sample of rabbit erythrocytes released 0.80 (+/- 0.13) microM ATP when focused to a cross-section of 3480 microm(2), while focusing the same sample to a smaller cross-section (1160 microm(2)) led to a release of 6.43 (+/- 0.40) microM ATP. In addition, two different inhibitors, diamide and glibenclamide, were used to ensure a lack of cell lysis. This approach can be used to examine a wide range of deformation forces in a high throughput fashion and will be of interest to researchers studying the mechanisms leading to vasodilation in the microvasculature.

  5. The Cutaneous Rabbit Revisited

    ERIC Educational Resources Information Center

    Flach, Rudiger; Haggard, Patrick

    2006-01-01

    In the cutaneous rabbit effect (CRE), a tactile event (so-called attractee tap) is mislocalized toward an adjacent attractor tap. The effect depends on the time interval between the taps. The authors delivered sequences of taps to the forearm and asked participants to report the location of one of the taps. The authors replicated the original CRE…

  6. Autoantibody Production in Rabbits

    PubMed Central

    Asherson, G. L.; Dumonde, D. C.

    1963-01-01

    The sera of rabbits injected with rat liver, kidney, heart, muscle, spleen and brain in Freund's complete adjuvant fixed complement with rabbit tissue. This complement-fixing activity was attributed to autoantibodies which were able to fix complement in vitro with the tissue of the rabbit in which they occurred. Absorption, gel diffusion and antibody and antigen titrations indicated that some of the anti-liver, anti-kidney, anti-heart, anti-muscle and anti-brain sera contained organ-specific autoantibody. The sera also contained autoantibody reacting with widely distributed antigen(s), which was relatively labile at 65°. The anti-kidney and anti-brain sera reacted with distinct antigens which were extracted from rabbit kidney and brain with a mixture of chloroform and methanol. The natural autoantibody of Kidd and Friedewald was usually labile at 65° and behaved like a macroglobulin on sucrose gradient centrifugation. Sera taken 1 week after immunization with rat tissue contained heat-labile macroglobulin antibody. However, sera taken 1 month after immunization also contained small molecular weight antibody which was stable at 65°. PMID:13965166

  7. 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

  8. 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.

  9. 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.

  10. 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.

  11. The relationship of the lunar regolith less than 10-microns fraction and agglutinates. II - Chemical composition of agglutinate glass as a test of the 'fusion of the finest fraction' /F3/ model

    NASA Technical Reports Server (NTRS)

    Walker, R. J.; Papike, J. J.

    1982-01-01

    Agglutinate glasses from nine Apollo soils have been studied using an automated electron microprobe technique in order to test the fusion of the finest fraction model proposed by Papike (1981). The nine average agglutinate glass compositions are compared with the calculated fused-soil-free compositions, the bulk compositions and the 90-20 micron fraction compositions of the soils in which they are found. It is found that the agglutinate glass data are consistent with the composition of most of the fractions finer than 10 microns, allowing for the volatile loss of K2O and Na2O; some inconsistencies that do arise may result from the degree of soil maturity and the amount of material finer than 10 microns. It is concluded that the fusion of the finest fraction model is a good first approximation of mechanisms affecting the formation of agglutinate glass.

  12. 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.

  13. Autoantibody production in rabbits

    PubMed Central

    Asherson, G. L.; Holborow, E. J.

    1966-01-01

    Rabbits received two injections of dead bacteria in Freund's complete adjuvant. One month later the sera of the rabbits were examined for autoantibodies against gut by indirect immunofluorescence using the rabbit's own stomach, ileum and colon taken at post mortem. Autoantibodies against colon were found in three out of seven rabbits injected with one particular strain of Escherichia coli O64 and in a few animals injected with other E. coli, Salmonella arizona, Proteus mirabilis, Klebsiella pneumoniae and Streptococcus faecalis. The antigen, with which the autoantibodies reacted, behaved like mucus and was detected in the colon and sometimes in the ileum and the stomach. Three patterns of staining were observed: (a) staining of the superficial mucosa of the colon with sparing of the deep glands; (b) staining of scattered groups of glands in the deepest part of the colon with sparing of the superficial glands (this pattern of staining was associated with staining of the superficial mucosa of the body of the stomach); and (c) staining of both the superficial and deep glands of the colon. None of the sera tested reacted with the bronchial or salivary glands. Polysaccharide preparations of the colon, but not the stomach, inhibited the reaction of the autoantibodies with colon in the sera tested. The amount of antigen needed to inhibit the basal staining was much greater than that needed to inhibit the superficial staining. It was concluded that rabbits may produce autoantibodies to colon and in some cases to ileum and stomach following the injection of certain dead bacteria in Freund's complete adjuvant. ImagesFIGS. 1-2FIG. 3FIG. 4FIG. 5FIG. 6 PMID:4956607

  14. MEMS reagent and sample handling procedure: Feasibility of viral antibody detection by passive immune agglutination

    NASA Technical Reports Server (NTRS)

    Bailey, G. D.; Tenoso, H. J.

    1975-01-01

    An attempt was made to develop a test requiring no preadsorption steps for the assessment of antibodies to rubella and mumps viruses using the passive immune agglutination (PIA) method. Both rubella and mumps antigens and antibodies were prepared. Direct PIA tests, using rubella antigen-coated beads, and indirect PIA tests, using rubella antibody-coated beads, were investigated. Attempts, using either method, were unsuccessful. Serum interference along with nonspecific agglutination of beads by the rubella antigen resulted in no specific response under the test conditions investigated. A new, highly sensitive approach, the enzyme immunoassay (EIA) test system, is recommended to overcome the nonspecificity. This system is a logical outgrowth of some of the solid phase work done on MEMS and represents the next generation tests system that can be directly applied to early disease detection and monitoring.

  15. Alpha mating type-specific expression of mutations leading to constitutive agglutinability in Saccharomyces cerevisiae.

    PubMed Central

    Doi, S; Yoshimura, M

    1985-01-01

    Two mutants of Saccharomyces cerevisiae have been isolated and characterized. The mutants were constitutively agglutinable at 36 degrees C, the temperature at which wild-type cells agglutinate only after induction by mating pheromone. The mutant cells had other properties specific for the normal alpha cell type, i.e., conjugation with a cells, response to a mating pheromone, and production of alpha mating pheromone. The two mutations, cag1 and cag2, were recessive and expressed only in alpha cells. cag1 is linked very closely to the MAT locus, but cag2 is unlinked to the MAT locus. These cag mutations complemented ste3-1. These results indicate that CAG genes are novel alpha-specific genes involved in the regulation of sex agglutinin synthesis. PMID:3881403

  16. Lead isotopic studies of lunar soils - Their bearing on the time scale of agglutinate formation

    NASA Technical Reports Server (NTRS)

    Church, S. E.; Tilton, G. R.; Chen, J. H.

    1976-01-01

    Fines (smaller than 75 microns) and bulk soil were studied to analyze loss of volatile lead; losses of the order of 10% to 30% radiogenic lead during the production of agglutinates are assessed. Lead isotope data from fine-agglutinate pairs are analyzed for information on the time scale of micrometeorite bombardment, from the chords generated by the data in concordia diagrams. Resulting mean lead loss ages were compared to spallogenic gas exposure ages for all samples. Labile parentless radiogenic Pb residing preferentially on or in the fines is viewed as possibly responsible for aberrant lead loss ages. Bulk soils plot above the concordia curve (in a field of excess radiogenic Pb) for all samples with anomalous ages.

  17. [Diagnostic value of agglutination reaction with buffered Brucella antigen stained with rose bengal].

    PubMed

    Chenchev, I; Khristoforov, L; Peshkov, I; Kostov, G; Mineva, I

    1978-01-01

    A specific buffered antigen has been obtained, employing a method developed by the authors, stained with Bengal rose and intended for performing a fast agglutination reaction to confirm brucellosis. The antigen produces a clear and demonstrative agglutination reaction with positive sera. Practically, the test is readily carried out, and can be made a routine both in every serologic laboratory and for investigations under field conditions. The reaction produced with this antigen can determine dependably the epizootic status on a farm or in the herd. The diagnostic value of the antigen is essential, especially with swine. Besides, it shows a wide a diagnostic scope for brucellosis with all species of animals (97--98 per cent).

  18. Enhancement of Leptospira hardjo agglutination titers in sheep and goat serum by heat inactivation.

    PubMed

    Malkin, K

    1984-04-01

    Heat inactivation of sheep serum samples resulted in the detection of an additional 9% reactors to Leptospira hardjo that were negative on the initial test of fresh samples. Treatment with EDTA gave results generally similar to heat inactivation suggesting that complement was responsible for the inhibition of agglutination. Tests on heat inactivated serum from experimentally infected sheep and goats revealed enhanced titers or reactions which were not detected in fresh serum.

  19. 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.

  20. 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.

  1. [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.

  2. Simplified spectraphotometric method for the detection of red blood cell agglutination.

    PubMed

    Ramasubramanian, Melur; Anthony, Steven; Lambert, Jeremy

    2008-08-01

    Human error is the most significant factor attributed to incompatible blood transfusions. A spectrophotometric approach to blood typing has been developed by examining the spectral slopes of dilute red blood cell (RBC) suspensions in saline, in the presence and absence of various antibodies, offering a technique for the quantitative determination of agglutination intensity [Transfusion39, 1051, 1999TRANAT0041-113210.1046/j.1537-2995.1999.39101051.x]. We offer direct theoretical prediction of the observed change in slope in the 660-1000 nm range through the use of the T-matrix approach and Lorenz-Mie theory for light scattering by dilute RBC suspensions. Following a numerical simulation using the T-matrix code, we present a simplified sensing method for detecting agglutination. The sensor design has been prototyped, fully characterized, and evaluated through a complete set of tests with over 60 RBC samples and compared with the full spectrophotometric method. The LED and photodiode pairs are found to successfully reproduce the spectroscopic determination of red blood cell agglutination.

  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. [Effect of extremely low frequency electromagnetic radiation and ultra-violet radiation on aggregation of thymocytes and erythrocytes].

    PubMed

    Roshchupkin, D I; Kramarenko, G G; Anosov, A K

    1996-01-01

    Electromagnetic radiation of superhigh frequencies (46.12 and 46.19 GHz, 0.3-1 mV/cm2) at an incident dose of about 12 kJ/m2 enhances the ability of isolated rabbit thymocytes for aggregation interaction with homologous erythrocytes. In the case of 46.19 GHz frequency, the stimulatory effect disappears as radiation dose in increased. A radiation of 46.12 GHz stimulates thymocytes also at high radiation doses. Superhigh-frequency radiation enhances the sensitivity of thymocytes to the damaging effect of UV radiation.

  5. D-penicillamine prevents ram sperm agglutination by reducing the disulphide bonds of a copper-binding sperm protein.

    PubMed

    Leahy, T; Rickard, J P; Aitken, R J; de Graaf, S P

    2016-05-01

    Head-to-head agglutination of ram spermatozoa is induced by dilution in the Tyrode's capacitation medium with albumin, lactate and pyruvate (TALP) and ameliorated by the addition of the thiol d-penicillamine (PEN). To better understand the association and disassociation of ram spermatozoa, we investigated the mechanism of action of PEN in perturbing sperm agglutination. PEN acts as a chelator of heavy metals, an antioxidant and a reducing agent. Chelation is not the main mechanism of action, as the broad-spectrum chelator ethylenediaminetetraacetic acid and the copper-specific chelator bathocuproinedisulfonic acid were inferior anti-agglutination agents compared with PEN. Oxidative stress is also an unlikely mechanism of sperm association, as PEN was significantly more effective in ameliorating agglutination than the antioxidants superoxide dismutase, ascorbic acid, α-tocopherol and catalase. Only the reducing agents cysteine and DL-dithiothreitol displayed similar levels of non-agglutinated spermatozoa at 0 h compared with PEN but were less effective after 3 h of incubation (37 °C). The addition of 10 µM Cu(2+) to 250 µM PEN + TALP caused a rapid reversion of the motile sperm population from a non-agglutinated state to an agglutinated state. Other heavy metals (cobalt, iron, manganese and zinc) did not provoke such a strong response. Together, these results indicate that PEN prevents sperm association by the reduction of disulphide bonds on a sperm membrane protein that binds copper. ADAM proteins are possible candidates, as targeted inhibition of the metalloproteinase domain significantly increased the percentage of motile, non-agglutinated spermatozoa (52.0% ± 7.8) compared with TALP alone (10.6% ± 6.1).

  6. From rabbit antibody repertoires to rabbit monoclonal antibodies

    PubMed Central

    Weber, Justus; Peng, Haiyong; Rader, Christoph

    2017-01-01

    In this review, we explain why and how rabbit monoclonal antibodies have become outstanding reagents for laboratory research and increasingly for diagnostic and therapeutic applications. Starting with the unique ontogeny of rabbit B cells that affords highly distinctive antibody repertoires rich in in vivo pruned binders of high diversity, affinity and specificity, we describe the generation of rabbit monoclonal antibodies by hybridoma technology, phage display and alternative methods, along with an account of successful humanization strategies. PMID:28336958

  7. Comparison of genomic and antimicrobial resistance features of latex agglutination test-positive and latex agglutination test-negative Staphylococcus aureus isolates causing bovine mastitis.

    PubMed

    Moser, A; Stephan, R; Corti, S; Johler, S

    2013-01-01

    The dairy industry suffers massive economic losses due to staphylococcal mastitis in cattle. The Staphaureux latex agglutination test (Oxoid, Basel, Switzerland) was reported to lead to negative results in 54% of bovine Staphylococcus aureus strains, and latex-negative strains are thought to be less virulent than Staphaurex latex-positive strains. However, comparative information on virulence and resistance profiles of these 2 groups of Staph. aureus is scarce. Our objective was to associate the latex agglutination phenotype of Staph. aureus strains isolated from bovine mastitis milk with data on clonal complexes, virulence genes, and antibiotic resistance to (1) determine the virulence profiles of the Staphaureux test positive and Staphaurex test negative groups, and (2) provide data needed to improve treatment of bovine mastitis and to identify potential vaccine targets. Seventy-eight Staph. aureus strains isolated from 78 cows on 57 Swiss farms were characterized. Latex agglutination was tested by Staphaureux kit, and resistance profiles were generated by disk diffusion. A DNA microarray was used to assign clonal complexes (CC) and to determine virulence and resistance gene profiles. By the Staphaureux test, 49% of the isolates were latex-positive and 51% were latex-negative. All latex-negative strains were assigned to CC151, whereas latex-positive strains were assigned to various clonal complexes, including CC97 (n=16), CC8 (n=10), CC479 (n=5), CC20 (n=4), CC7 (n=1), CC9 (n=1), and CC45 (n=1). Although the latex-negative isolates were susceptible to all antimicrobial agents tested, 24% of latex-positive isolates were classified as intermediate with regard to cefalexin-kanamycin and 13% were resistant to both ampicillin and penicillin. Microarray profiles of latex-negative isolates were highly similar, but differed largely from those of latex-positive isolates. Although the latex-negative group lacked several enterotoxin genes and sak, it exhibited significantly

  8. 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

  9. 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.

  10. 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)

  11. 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.

  12. 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.

  13. 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.

  14. Rabbits killing birds revisited.

    PubMed

    Zhang, Jimin; Fan, Meng; Kuang, Yang

    2006-09-01

    We formulate and study a three-species population model consisting of an endemic prey (bird), an alien prey (rabbit) and an alien predator (cat). Our model overcomes several model construction problems in existing models. Moreover, our model generates richer, more reasonable and realistic dynamics. We explore the possible control strategies to save or restore the bird by controlling or eliminating the rabbit or the cat when the bird is endangered. We confirm the existence of the hyperpredation phenomenon, which is a big potential threat to most endemic prey. Specifically, we show that, in an endemic prey-alien prey-alien predator system, eradication of introduced predators such as the cat alone is not always the best solution to protect endemic insular prey since predator control may fail to protect the indigenous prey when the control of the introduced prey is not carried out simultaneously.

  15. Rabbit renotropic system

    SciTech Connect

    Areas, J.; Yun, G.C.; Rahmat, J.; Gersten, D.; Goel, R.; Preuss, H.G.

    1988-04-01

    Elevated levels of a specific renal growth factor, renotropin, have been associated with spontaneous hypertension. To examine this association more closely, we have undertaken the development of a better assay system to characterize and purify renotropin. Sera from rabbits prior to operation (control) and at a specified time after unilateral nephrectomy (uni) were examined for renotropic activity. Comparing the effects of uni to control sera in the same rabbit, significant stimulation of 3H-thymidine incorporation into the DNA of primary rabbit kidney cultures incubated in D-valine medium to eliminate fibroblast growth was noted: at 3 days postoperatively 73% (n = 13), at 7 days 103% (n = 39), at 10 days 130% (n = 31), at 21 days 101% (n = 24), at 42 days 89% (n = 13). All values were at least P less than 0.01. The stimulatory properties were dose-dependent but reached a plateau at high serum concentrations. Comparing CPM/mg protein in uni/control in different concentrations of sera 7 days postoperatively, uni versus control were 67/44 at 5% v/v, 139/72 at 10% v/v, 261/161 at 20% v/v, and 243/136 at 40% v/v. The renotropic effect of uni sera remained after dialysis in incubation medium and after sera were heated in boiling water for 5 minutes. Renal extracts obtained from growing kidneys 7 days postnephrectomy augmented renotropic activity. Atrial natriuretic factor, ouabain, PGF2 alpha, PGE1, and cAMP did not possess renotropic activity. We conclude that the primary rabbit kidney culture assay for renotropin is highly sensitive and will be an important tool to comprehend the role of renotropin in the pathogenesis of hypertension.

  16. 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.

  17. 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.

  18. 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

  19. 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.

  20. 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.

  1. Erythrocyte antigen and reticulocyte engraftment after allogeneic hematopoietic stem cell transplantation.

    PubMed

    Yildirim, I; Ozer, Y; Yüksel, M K; Arat, M; Arslan, O

    2004-08-01

    The aim of this study was to study the usefulness of erythrocyte antigen (EA) measurement to study engraftment after allogeneic HSCT. In all, 31 consecutive patients receiving HLA-identical bone marrow (BM) (n=13) or peripheral blood stem cells (n=18) were investigated. Apart from the ABO group, 15 EAs representing six minor blood groups were followed by the simple tube agglutination technique. A total of 20 (64.5%) patients received ABO-identical, eight (25.8%) received ABO minor and three (9.7%) received ABO major mismatched grafts. In all, 29 patients were followed for a median of 12 (6-16) months; 65% of the patients expressed donor type EA 1 month and almost all did so 6 months after transplant. Reticulocyte engraftment was significantly shorter than EA engraftment (median 18 vs 35 days) (P=0.001). Patients who received PB stem cells showed significantly faster EA and reticulocyte engraftment than patients who received BM stem cells (P=0.038 and 0.025). ABO compatibility did not have an impact on reticulocyte and EA engraftment (P=0.4 and 0.55). The earliest donor type EA detected was from the Rh and Kidd system. These data suggest that EA and reticulocyte assays are useful in monitoring engraftment.

  2. The Osmotically Functional Water Content of the Human Erythrocyte

    PubMed Central

    LeFevre, Paul G.

    1964-01-01

    Experiments were directed toward estimation of the magnitude of error incurred by the presumption of idealized osmometric behavior in the author's recent studies of monosaccharide transport through the human erythrocyte membrane. Thick suspensions of washed cells in isotonic buffered balanced salt medium were mixed in fixed proportions with varying dilutions of a concentrate of either (a) the mixed chlorides of the medium, or (b) glucose in the isotonic medium, and the resultant freezing point and hematocrit values determined. The form of the responses in the tonicity and the cell volume, as functions of the variable dilution of sugar or salts, conformed consistently with relations derived from the classical van't Hoff-Boyle-Mariotte pressure-volume relation. However, the effective cell water contents appeared substantially less than the weight lost in conventional drying, and varied somewhat according to the index used: expressed as grams of H2O per milliliter of cells at isotonic volume, the cell water implied by the hematocrit behavior was 0.614 ± 0.015 (SD); by the salt tonicity response, 0.565 ± 0.027; by the immediate glucose tonicity response, 0.562 ± 0.044; and by the equilibrium glucose tonicities, 0.589 ± 0.043. Olmstead's reports of gross deviation from the van't Hoff relation, in the rabbit red cell's responses to tonicity displacement, are attributed primarily to a systematic aberration in his method of data analysis, the observations themselves agreeing substantially with the present findings. PMID:14100971

  3. Herpes simplex virus type 1-induced hemagglutination: glycoprotein C mediates virus binding to erythrocyte surface heparan sulfate.

    PubMed Central

    Trybala, E; Svennerholm, B; Bergström, T; Olofsson, S; Jeansson, S; Goodman, J L

    1993-01-01

    We recently reported that herpes simplex virus type 1 (HSV-1) can cause agglutination of murine erythrocytes (E. Trybala, Z. Larski, and J. Wisniewski, Arch. Virol. 113:89-94, 1990). We now demonstrate that the mechanism of this hemagglutination is glycoprotein C-mediated binding of virus to heparan sulfate moieties at the surface of erythrocytes. Hemagglutination was found to be a common property of all gC-expressing laboratory strains and clinical isolates of HSV-1 tested. Mutants of HSV-1 deficient in glycoprotein C caused no specific hemagglutination, whereas their derivatives transfected with a functional gC-1 gene, thus reconstituting gC expression, regained full hemagglutinating activity. Hemagglutination activity was inhibited by antibodies against gC-1 but not by antibodies with specificity for glycoproteins gB, gD, or gE or by murine antiserum raised against the MP strain of HSV-1, which is gC deficient. Finally, purified gC-1 protein, like whole HSV-1 virions, showed high hemagglutinating activity which was inhibited by heparan sulfate and/or heparin and was completely prevented by pretreatment of erythrocytes with heparitinase, providing evidence that gC-1 mediates hemagglutination by binding to heparan sulfate at the cell surface. Thus, HSV-1-induced hemagglutination is gC-1 dependent and resembles the recently proposed mechanism by which HSV-1 attaches to surface heparans on susceptible cells, providing a simple model for initial events in the virus-cell interaction. Images PMID:8382294

  4. Effects of corticosteroids and splenectomy on the immune clearance and destruction of erythrocytes.

    PubMed

    Atkinson, J P; Schreiber, A D; Frank, M M

    1973-06-01

    Corticosteroids and splenectomy constitute two important therapeutic modalities in the treatment of autoimmune hemolytic anemia. Each of these may affect both the rate of synthesis of autoantibody and the clearance of antibody sensitized cells. The latter possibility has been examined in an experimental model which allows evaluation of the role of antibody and complement in the immune clearance of erythrocytes in molecular terms. The in vivo clearance of (51)Cr-labeled guinea pig erythrocytes sensitized with purified rabbit IgG or IgM antibody to produce a known number of complement-fixing sites per cell was studied. Corticosteroid therapy increased the survival of both IgG and IgM sensitized erythrocytes by decreasing sequestration in the reticuloendothelial system (RES). 5 days of therapy prior to injection of antibody coated cells were required for a maximal effect. It appeared that the RES of cortisone-treated animals had a lowered sensitivity to erythrocytes coated with antibody and complement and the cells were removed as though they were coated with fewer complement sites/cell. The general pattern and kinetics of clearance and the localization of sequestered cells were not modified by corticosteroids. As the number of IgG C1-fixing sites was increased, the difference betwen cortisone treated and control animals was less marked. Splenectomy led to an increased survival of IgG-coated cells and a sixfold increase in IgG C1-fixing sites was necessary in order to obtain similar rates of clearance in splenectomized and control animals. The liver was responsible for this much less efficient clearance of cells in splenectomized animals: the clearance pattern was typical of that noted for IgG. No effect at all was noted on the clearance of IgM sensitized cells in splenectomized animals. These experiments clearly demonstrate that both corticosteroid therapy and splenectomy act to decrease the in vivo clearance of IgG-sensitized cells: only corticosteroids alter the

  5. Effects of Corticosteroids and Splenectomy on the Immune Clearance and Destruction of Erythrocytes

    PubMed Central

    Atkinson, John P.; Schreiber, Alan D.; Frank, Michael M.

    1973-01-01

    Corticosteroids and splenectomy constitute two important therapeutic modalities in the treatment of autoimmune hemolytic anemia. Each of these may affect both the rate of synthesis of autoantibody and the clearance of antibody sensitized cells. The latter possibility has been examined in an experimental model which allows evaluation of the role of antibody and complement in the immune clearance of erythrocytes in molecular terms. The in vivo clearance of 51Cr-labeled guinea pig erythrocytes sensitized with purified rabbit IgG or IgM antibody to produce a known number of complement-fixing sites per cell was studied. Corticosteroid therapy increased the survival of both IgG and IgM sensitized erythrocytes by decreasing sequestration in the reticuloendothelial system (RES). 5 days of therapy prior to injection of antibody coated cells were required for a maximal effect. It appeared that the RES of cortisone-treated animals had a lowered sensitivity to erythrocytes coated with antibody and complement and the cells were removed as though they were coated with fewer complement sites/cell. The general pattern and kinetics of clearance and the localization of sequestered cells were not modified by corticosteroids. As the number of IgG C1-fixing sites was increased, the difference betwen cortisone treated and control animals was less marked. Splenectomy led to an increased survival of IgG-coated cells and a sixfold increase in IgG C1-fixing sites was necessary in order to obtain similar rates of clearance in splenectomized and control animals. The liver was responsible for this much less efficient clearance of cells in splenectomized animals: the clearance pattern was typical of that noted for IgG. No effect at all was noted on the clearance of IgM sensitized cells in splenectomized animals. These experiments clearly demonstrate that both corticosteroid therapy and splenectomy act to decrease the in vivo clearance of IgG-sensitized cells: only corticosteroids alter the

  6. Nanoscale Mineralogy and Composition of Experimental Regolith Agglutinates Produced under Asteroidal Impact Conditions

    NASA Technical Reports Server (NTRS)

    Christoffersen, Roy; Cintala, M. J.; Keller, L. P.; See, T. H.; Horz, F.

    2013-01-01

    On the Moon, the energetics of smaller impactors and the physical/chemical characteristics of the granular regolith target combine to form a key product of lunar space weathering: chemically reduced shock melts containing optically-active nanophase Fe metal grains (npFe0) [1]. In addition to forming the optically dark glassy matrix phase in lunar agglutinitic soil particles [1], these shock melts are becoming increasingly recognized for their contribution to optically active patina coatings on a wide range of exposed rock and grain surfaces in the lunar regolith [2]. In applying the lessons of lunar space weathering to asteroids, the potential similarities and differences in regolith-hosted shock melts on the Moon compared to those on asteroids has become a topic of increasing interest [3,4]. In a series of impact experiments performed at velocities applicable to the asteroid belt [5], Horz et al. [6] and See and Horz [7] have previously shown that repeated impacts into a gabbroic regolith analog target can produce melt-welded grain aggregates morphologically very similar to lunar agglutinates [6,7]. Although these agglutinate-like particles were extensively analyzed by electron microprobe and scanning electron microscopy (SEM) as part of the original study [7], a microstructural and compositional comparison of these aggregates to lunar soil agglutinates at sub-micron scales has yet to be made. To close this gap, we characterized a representative set of these aggregates using a JEOL 7600 field-emission scanning electron microscope (FE-SEM), and JEOL 2500SE field-emission scanning transmission electron microscope (FE-STEM) both optimized for energy dispersive X-ray spectroscopy (EDX) compositional spectrum imaging at respective analytical spatial resolutions of 0.5 to 1 micron, and 2 to 4 nm.

  7. Antibodies to spermatozoa. I. A new macroscopic agglutination technique for their detection, using immotile sperm

    PubMed Central

    Shulman, S.; Hekman, Annemarie

    1971-01-01

    An agglutination procedure, especially useful for guinea-pig sperm, has been developed. The preferred conditions are as follows: epididymal spermatozoa, in an immotile condition, unwashed or diluted and centrifuged once, at a concentration of 10×106 per ml diluted in phosphate-buffered saline, are incubated with antiserum samples in capillary tubes at 4°C for 1 or 2 hr of observation. This procedure has readily allowed us to detect the presence of antibodies against epididymal spermatozoa. The test has given results that are quite parallel to those obtained by the conventional Kibrick method. ImagesFig. 1Fig. 2 PMID:5570676

  8. [Comparative measurement of antithrombin III by latex agglutination and radial immunodiffusion in patients with peritonitis].

    PubMed

    Miagkova, M A; Aleshkin, A V; Abramenko, T V; Savitskaia, Iu A; Aleshkin, V A

    1997-01-01

    A highly sensitive and rapid method, based on latex agglutination, has been developed for measuring antithrombin III (AT III) in the blood serum of patients and donors. The sensitivity of analysis is 0.6 microgram/ml, time 2 to 3 min. The method is simple, requires no sophisticated equipment, and may be used under field conditions. The results are assessed visually. Immunochemical reagents have been synthesized for the method: latex conjugates and specific antibodies to AT III. The method was tried in patients with peritonitis. An additional criterion for diagnosing the respiratory distress syndrome of adults in this patient population has been developed.

  9. Diabetic erythrocytes test by correlation coefficient.

    PubMed

    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.

  10. 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

  11. 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.

  12. 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.

  13. Erythrocyte receptor (CD2)-bearing T lymphocytes are affected by diet in experimental pulmonary tuberculosis.

    PubMed Central

    Bartow, R A; McMurray, D N

    1990-01-01

    Outbred (Hartley) and inbred (strain 2 and 13) guinea pigs were vaccinated with Mycobacterium bovis BCG and placed on isocaloric, purified diets containing either 10 or 30% ovalbumin or on commercial guinea pig chow. Six weeks later, the animals were challenged by the respiratory route with virulent M. tuberculosis H37Rv. At intervals postchallenge, groups were tuberculin tested and sacrificed. Thymus-dependent (T) lymphocytes were enumerated either by rosette formation with rabbit erythrocytes or by the indirect immunofluorescence assay (IFA) with a guinea pig pan-T-cell monoclonal antibody, 8BE6. Protein-deficient guinea pigs of all three strains had significantly fewer erythrocyte rosette-forming (CD2+) T cells in the peripheral blood, and malnourished strain 2 and Hartley guinea pigs exhibited reduced levels of CD2+ T cells in the thymus. In contrast, animals of all three strains fed the low-protein diet harbored more CD2+ T cells in the bronchotracheal lymph nodes than did their control-fed counterparts. A larger proportion of lymphocytes from the blood and lymph nodes of all three strains were IFA positive than formed erythrocyte rosettes regardless of diet treatment. Diet had no effect on IFA-positive lymphocytes in those organs. Protein deficiency is associated with significant alterations in the number and/or distribution of T lymphocytes expressing functional CD2-receptors in BCG-vaccinated animals exposed to virulent mycobacteria by the pulmonary route. These alterations may contribute to the reduction in BCG vaccine efficacy observed in this model. PMID:1971259

  14. 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

  15. 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.

  16. A Possible Role for Agglutinated Foraminifers in the Growth of Deep-Water Coral Bioherms

    NASA Astrophysics Data System (ADS)

    Messing, C. G.; Reed, J. K.; Brooke, S. D.

    2008-05-01

    Exploration of deep-water bioherms dominated by the scleractinian corals Lophelia pertusa and Enallopsammia profunda along the east coast of Florida in ~400-800 m depth reveals an often dense and rich assemblage of small (~1-30 mm) epifauna on dead coral branches, which is often dominated by agglutinated astrorhizacean foraminifers accompanied by thecate and athecate hydroids, sponges, stylasterids, anemones and barnacles. The dominant agglutinated foraminifer is an arborescent form up to 15 mm tall, consisting of a basal tube that gives rise to branchlets of successively decreasing diameter and thickly coated with fine-grained material including coccoliths and diatom frustules. The large numbers of foraminifers generate an enormous adhesive, sediment-trapping surface area and may represent an important accelerated route for sediment deposition and bioherm growth relative to baffling of suspended sediment particles by the coral branches themselves. These foraminifers also occur on still living coral, suggesting that they may either contribute to coral death or invade stressed colonies. They may thus be responsible for or contribute to the small percent of living corals observed in many of these habitats. Other epifauna appear to colonize after the coral has died.

  17. Ethanol induces human red cell shape transformations and enhanced ligand-mediated agglutinability

    SciTech Connect

    Weinstein, R.S.; McLawhon, R.W.; Marikovsky, Y.

    1986-03-01

    Ethanol concentrations are markedly elevated in rat stomach wall when ulcerogenic doses of 100 % ethanol (2 ml for 5 to 10 minutes) are instilled in rat gastric lumen. The authors observed that red cells in gastric mucosal postcapillary venules become spiculated and interadherent under these conditions. The authors have now studied this phenomenon in vitro using washing human red cells. Concentrations of high grade ethanol ranging from 2 to 10% (v/v) in physiological buffered saline (pH 7.3) without Ca/sup + +/ or Mg/sup + +/ at 25/sup 0/C rapidly transformed human red cells into spiculated forms. 2% ethanol transformed human red cells into disco-echinocytes in 15 min. whereas 10% ethanol transformed red blood cells into echinocytes within 3 min. Washing out of ethanol at 1 hour reverted the echinocytes into discocytes. However, following 3 hours of incubation in 10% ethanol washing out of ethanol produced stomatocytes. The ethanol-induced echinocytic shape transformations were accompanied by a dose-related increase in red cell agglutinability with poly-L-lysine or the plant lectin wheat germ agglutinin. The enhanced agglutinability was reversed by restoring the red cell shape changes and alterations in surface properties may play a role in the pathogenesis of ethanol-induced gastric ulcers.

  18. Comparison of PCR, Wright agglutination test and blood culture for diagnosis of brucellosis in suspected patients.

    PubMed

    Hekmatimoghaddam, Seyedhosssein; Sadeh, Maryam; Khalili, Mohammad Bagher; Mollaabedin, Mansour; Sazmand, Alireza

    2013-11-15

    Brucellosis has long been prevalent in Iran, with considerable medical and economic importance. Timely diagnosis is needed for early management and effective prevention of its consequences in human beings and animals. Current diagnostic methods impose peculiar challenges in terms of analytical method performance. This study compares diagnostic sensitivity, specificity, predictive Value of Positive (PVP) and Predictive Value of Negative (PVN) for Polymerase Chain Reaction (PCR), Wright agglutination test and blood culture used for patients suspected of brucellosis. In 120 patients clinically suspected of brucellosis and referred by physicians to the Yazd central Medical Laboratory, some relevant demographic, occupational, nutritional and clinical data were collected. Also, venous blood samples were drawn for diagnosis of brucellosis using PCR, Wright agglutination test and blood culture techniques. The most frequent symptom of patients was arthralgia (82 cases, 68.3%). PCR was positive in 25 cases (20.8%), wright test in 21 patients (17.5%) and blood culture in 6 cases (5%). In 20 out of 21 wright-positive cases, PCR was positive and all of the culture-positive patients had positive PCR. Sensitivity, specificity, PVP and PVN of blood culture compared to PCR (as the gold standard test) were 24, 100, 100 and 86%, respectively, but the above parameters when PCR is compared with blood culture (as gold standard) were 100, 83, 24 and 95%, respectively. PCR has better analytical performances than blood culture for diagnosis of brucellosis and is suitable for confirmation of Wright-positive cases.

  19. Amyloidogenic amyloid-β-peptide variants induce microbial agglutination and exert antimicrobial activity

    PubMed Central

    Spitzer, Philipp; Condic, Mateja; Herrmann, Martin; Oberstein, Timo Jan; Scharin-Mehlmann, Marina; Gilbert, Daniel F.; Friedrich, Oliver; Grömer, Teja; Kornhuber, Johannes; Lang, Roland; Maler, Juan Manuel

    2016-01-01

    Amyloid-β (Aβ) peptides are the main components of the plaques found in the brains of patients with Alzheimer’s disease. However, Aβ peptides are also detectable in secretory compartments and peripheral blood contains a complex mixture of more than 40 different modified and/or N- and C-terminally truncated Aβ peptides. Recently, anti-infective properties of Aβ peptides have been reported. Here, we investigated the interaction of Aβ peptides of different lengths with various bacterial strains and the yeast Candida albicans. The amyloidogenic peptides Aβ1-42, Aβ2-42, and Aβ3p-42 but not the non-amyloidogenic peptides Aβ1-40 and Aβ2-40 bound to microbial surfaces. As observed by immunocytochemistry, scanning electron microscopy and Gram staining, treatment of several bacterial strains and Candida albicans with Aβ peptide variants ending at position 42 (Aβx-42) caused the formation of large agglutinates. These aggregates were not detected after incubation with Aβx-40. Furthermore, Aβx-42 exerted an antimicrobial activity on all tested pathogens, killing up to 80% of microorganisms within 6 h. Aβ1-40 only had a moderate antimicrobial activity against C. albicans. Agglutination of Aβ1-42 was accelerated in the presence of microorganisms. These data demonstrate that the amyloidogenic Aβx-42 variants have antimicrobial activity and may therefore act as antimicrobial peptides in the immune system. PMID:27624303

  20. 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.

  1. High Bacterial Agglutination Activity in a Single-CRD C-Type Lectin from Spodoptera exigua (Lepidoptera: Noctuidae)

    PubMed Central

    Gasmi, Leila; Ferré, Juan; Herrero, Salvador

    2017-01-01

    Lectins are carbohydrate-interacting proteins that play a pivotal role in multiple physiological and developmental aspects of all organisms. They can specifically interact with different bacterial and viral pathogens through carbohydrate-recognition domains (CRD). In addition, lectins are also of biotechnological interest because of their potential use as biosensors for capturing and identifying bacterial species. In this work, three C-type lectins from the Lepidoptera Spodoptera exigua were produced as recombinant proteins and their bacterial agglutination properties were characterized. The lowest protein concentration producing bacterial agglutination against a panel of different Gram+ and Gram− as well as their carbohydrate binding specificities was determined for the three lectins. One of these lectins, BLL2, was able to agglutinate cells from a broad range of bacterial species at an extremely low concentration, becoming a very interesting protein to be used as a biosensor or for other biotechnological applications involving bacterial capture. PMID:28257054

  2. High Bacterial Agglutination Activity in a Single-CRD C-Type Lectin from Spodoptera exigua (Lepidoptera: Noctuidae).

    PubMed

    Gasmi, Leila; Ferré, Juan; Herrero, Salvador

    2017-03-01

    Lectins are carbohydrate-interacting proteins that play a pivotal role in multiple physiological and developmental aspects of all organisms. They can specifically interact with different bacterial and viral pathogens through carbohydrate-recognition domains (CRD). In addition, lectins are also of biotechnological interest because of their potential use as biosensors for capturing and identifying bacterial species. In this work, three C-type lectins from the Lepidoptera Spodoptera exigua were produced as recombinant proteins and their bacterial agglutination properties were characterized. The lowest protein concentration producing bacterial agglutination against a panel of different Gram+ and Gram- as well as their carbohydrate binding specificities was determined for the three lectins. One of these lectins, BLL2, was able to agglutinate cells from a broad range of bacterial species at an extremely low concentration, becoming a very interesting protein to be used as a biosensor or for other biotechnological applications involving bacterial capture.

  3. In vitro effects of 50 Hz magnetic fields on oxidatively damaged rabbit red blood cells

    SciTech Connect

    Fiorani, M.; Biagiarelli, B.; Vetrano, F.; Guidi, G.; Dacha, M.; Stocchi, V.

    1997-05-01

    The aim of this study was to investigate the effects of 50 Hz magnetic fields on rabbit red blood cells (RBCs) that were exposed simultaneously to the action of an oxygen radical-generating system, Fe(II)/ascorbate. Previous data obtained in the authors` laboratory showed that the exposure of rabbit erythrocytes or reticulocytes to Fe(II)/ascorbate induces hexokinase inactivation, whereas the other glycolytic enzymes do not show any decay. The authors also observed depletion of reduced glutathione (GSH) content with a concomitant intracellular and extracellular increase in oxidized glutathione (GSSG) and a decrease in energy charge. In this work, they investigated whether 50 Hz magnetic fields could influence the intracellular impairments that occur when erythrocytes or reticulocytes are exposed to this oxidant system, namely, inactivation of hexokinase activity, GSH depletion, a change in energy charge, and hemoglobin oxidation. The results obtained indicate that a 0.5 mT magnetic field had no effect on intact RBCs, whereas it increased the damage in an oxidatively stressed erythrocyte system. In fact, exposure of intact erythrocytes incubated with Fe(II)/ascorbate to a 0.5 mT magnetic field induced a significant further decay in hexokinase activity as well as a twofold increase in methemoglobin production compared with RBCs that were exposed to the oxidant system alone. Although further studies will be needed to determine the physiological implications of these data, the results reported in this study demonstrate that the effects of the magnetic fields investigated are able to potentiate the cellular damage induced in vitro by oxidizing agents.

  4. 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.

  5. 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.

  6. [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.

  7. 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.

  8. 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.

  9. Identification of serum component involved in generation of neo-lectin with agglutinating and phenoloxidase activities in human serum.

    PubMed

    Manikandan, Beulaja; Ramar, Manikandan; Munusamy, Arumugam

    2014-01-01

    Human serum albumin (HSA) was identified as the component involved in generation of neo-lectin molecules with both lectin and phenoloxidase activities. Pronase treated HSA was able to agglutinate hen RBC and oxidize hydroquinone. Sodium dodecyl sulphate (SDS) treated HSA agglutinated both hen and sheep RBC as well as oxidized dopamine. The hemagglutinating activities of pronase/SDS treated HSA observed against hen RBC were dosimetric. The oxidation of pronase/SDS treated HSA with hydroquinone/dopamine, respectively, was inhibitable by inhibitors of phenoloxidase, namely, phenylthiourea and tropolone. Very low concentrations of HSA could generate these humoral neo-lectin molecules.

  10. Preliminary observations on the use of latex agglutination test for the detection of mastitis due to Streptococcus agalactiae in cows.

    PubMed Central

    Daniel, R C; Barnum, D A

    1986-01-01

    A commercial latex agglutination test for the detection of Group B streptococcal antigens was used to detect infection due to Streptococcus agalactiae in whey of bovine milk samples. Fifteen out of 17 known infections were detected, but it was necessary to incubate the wheys at 37 degrees C for 18 hours in nine of the samples. It was found that the latex agglutination test could detect Group streptococcal carbohydrate antigens in whey samples from artificially infected quarters from one to four days after failure to detect the organism on culture or after antibiotic therapy of the affected quarter. PMID:3527389

  11. Oculoscopy in Rabbits and Rodents.

    PubMed

    Jekl, Vladimir; Hauptman, Karel; Knotek, Zdenek

    2015-09-01

    Ophthalmic diseases are common in rabbits and rodents. Fast and definitive diagnosis is imperative for successful treatment of ocular diseases. Ophthalmic examination in rabbits and rodents can be challenging. Oculoscopy offers great magnification for the examination of the ocular structures in such animals, including the evaluation of cornea, anterior eye chamber, limbus, iris, lens, and retina. To date, oculoscopy has been described only sporadically and/or under experimental conditions. This article describes the oculoscopy technique, normal and abnormal ocular findings, and the most common eye disorders diagnosed with the aid of endoscopy in rabbits and rodents.

  12. 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.

  13. 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

  14. 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

  15. Urethral healing in rabbits.

    PubMed

    Scherz, H C; Kaplan, G W; Boychuk, D I; Landa, H M; Haghighi, P

    1992-08-01

    We studied urethral healing in New Zealand white rabbits by histological examination after insult (urethral catheter) or injury (urethrotomy) specifically for acute and chronic inflammation, fibrosis, fistulas, squamous metaplasia, foreign body giant cells and urethral dilatation. Urethral catheterization resulted in increased inflammation and fibrosis compared to noncatheterized animals. Skin closure techniques and materials resulted in an inflammatory response that may extend to and involve the urethra. Minor differences in suture size were not an important variable but the persistence of suture material may have a role in the degree of inflammation and the formation of foreign body giant cells. Transepithelial closure techniques drag epithelial cells into subcutaneous tissues and may predispose to fistula formation.

  16. 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

  17. 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.

  18. 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.

  19. 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.

  20. The cottontail rabbits of Virginia

    USGS Publications Warehouse

    Llewellyn, L.M.; Handley, C.O.

    1945-01-01

    Five races of cottontail rabbits belonging to three species occur in Virginia. One of them, the Mearns cottontail (Sylvilagus floridanus mearnsi), is reported here for the first time. It occurs in six southwestern counties of the state, while the eastern cottontail (S. f. mallurus) occurs in the remainder of the state with the exception of Smith and Fishermans islands off the eastern coast of Cape Charles, where it is replaced by Hitchens cottontail (S. f. hitchensi). The New England cottontail (S. transitionalis) is found on the higher mountain peaks, above 3000 feet, and the swamp rabbit (S. palustris) occurs in the Dismal Swamp region of southeastern Virginia.....The height of the breeding season for the eastern cottontail in Virginia is March and April, but breeding continues through the entire year except in December and January. The average litter size based on embryo counts was 4.7. The sex ratio of 234 specimens from all parts of the state, taken mostly in the December to February period, was 53 males to 47 females. That of a group of 145 rabbits live-trapped at Blacksburg during February and Marchwas 58 males to 42 females. The figures show that males are more active than females during the winter months, and therefore are more easily taken then....In transplanting cottontails from one section of the state to another, it is recommended that only cottontails of the same race as those originally present in the region being restocked be released there....Tularemia is not a common disease among rabbits in Virginia, but the rabbit ticks are often carriers of the disease and may transmit it to rabbits. Rabbit ticks are also found to be carriers of Rocky Mountain fever and American Q. fever. After the ticks drop off the rabbits to hibernate in the ground, which is likely to occur during mid-winter in Virginia, there is relatively little danger of humans contracting tularemia by contact with rabbits. Present laws in Virginia which prohibit rabbit hunting until the

  1. 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.

  2. An experimental investigation of agglutinate melting mechanisms - Shocked mixtures of sodium and potassium feldspars

    NASA Technical Reports Server (NTRS)

    Simon, S. B.; Papike, J. J.; Horz, F.; See, T. H.

    1985-01-01

    The results of an experiment designed to test the validity of the model for agglutinate formation involving fusion of the finest fraction or F3 are reported. Impact glasses were formed from various mixes of orthoclase and albite powders, which were used as analogs for soils with chemically constrasting coarse and fine fractions. The results showed that the single most important factor displacing the composition of a small-scale impact melt from the bulk composition of the source regolith is the fractionated composition of the finest soil fraction. Volatile loss and the amount of melting, which in turn are determined by the degree of shock, are also important. As predicted by the model, the lower pressure melts are the most fractionated, and higher pressure is accompanied by increased melting causing glass compositions to approach the bulk. In general, the systematics predicted by the model are observed; the model appears to be valid.

  3. Latex agglutination test for detecting feline panleukopenia virus, canine parvovirus, and parvoviruses of fur animals.

    PubMed Central

    Veijalainen, P M; Neuvonen, E; Niskanen, A; Juokslahti, T

    1986-01-01

    A latex agglutination (LA) test for the detection of parvoviruses of fur animals, cats, and dogs was developed, and its sensitivity and specificity were compared with those of hemagglutination (HA) and the enzyme-linked immunosorbent assay (ELISA). Tissue culture isolation was used to confirm the specificity results. Fecal samples from various sources were tested, including specimens from raccoon dogs and mink which were experimentally infected with parvoviruses by oral exposure. LA compared favorably with the other tests. The ELISA was the most sensitive. When it was considered as a reference test, the corresponding sensitivities for HA and LA were 96 and 91%, respectively. The specificities were 93% for the ELISA, 95% for the HA test, and 92% for the LA test. LA seems to be a suitable technique for screening animals in the field and in laboratories in which sophisticated techniques are not available. PMID:3007568

  4. Case report: false negative serum cryptococcal latex agglutination test in a patient with disseminated cryptococcal disease.

    PubMed

    Navabi, Nazlee; Montebatsi, Milton; Scott, Michelle; Gluckman, Stephen J; Reid, Michael J A

    2015-01-01

    A case of false-negative serum latex agglutination cryptococcal antigen (CRAG) test in a 45-year-old HIV-positive male with Cryptococcus-positive culture is described. The patient was presented to a hospital in Botswana, with breathlessness and a diffuse papular rash. His CD4 count was 25 cells/μL. Despite the suspicion for disseminated cryptococcal disease, an initial serum CRAG latex test was negative. Results of subsequent Indian ink staining, culture of cerebrospinal fluid and skin scrapings, and serum lateral flow immunoassay (LFA) were all positive for Cryptococcus neoformans. There are several possible explanations for the false-negative CRAG latex test. Given the positive LFA result, we speculate that disease may have been caused by Cryptococcus gattii, which is estimated to be responsible for between 15% and 30% of all cryptococcal diseases in Botswana. Reduced sensitivity of CRAG latex assays for detecting C gattii may lead to underdiagnosis of cryptococcal infection.

  5. Evaluation of a commercial latex agglutination test for rapid detection of Salmonella in fecal samples.

    PubMed

    Bänffer, J R; van Zwol-Saarloos, J A; Broere, L J

    1993-08-01

    A latex agglutination test for the detection of salmonella in feces was evaluated in comparison to direct culture and enriched culture using both artificially inoculated samples and clinical samples. In the samples inoculated artificially with different concentrations of salmonella (10(1) to 10(5) per gram) the enriched culture performed better only at the 10(2) level in 0.4 g samples, whereas the latex test performed as well as the enriched culture at all levels in 4 g samples. In the tests using clinical samples, there was no significant difference between results of the latex test performed in 2283 samples and the enriched culture performed in 2072 samples. The sensitivity, specificity and negative and positive predictive values of the latex test were 88.2%, 98%, 97.5% and 63% respectively. The test provided results rapidly but yielded a number of false positive results.

  6. Prevalence of agglutinating antibodies to Toxoplasma gondii and Sarcocystis neurona in beavers (Castor canadensis) from Massachusetts.

    PubMed

    Jordan, Carly N; Kaur, Taranjit; Koenen, Kiana; DeStefano, Stephen; Zajac, Anne M; Lindsay, David S

    2005-10-01

    The present study examined the seroprevalence of Toxoplasma gondii and Sarcocystis neurona in a population of beavers (Castor canadensis) from Massachusetts. Sixty-two blood samples were collected during the field seasons over 3 consecutive years from different animals. Blood was collected onto filter paper and shipped to the Department of Biomedical Sciences, Virginia Tech, Blacksburg, Virginia, for parasite testing. The samples were tested at dilutions of 1:25, 1:50, and 1:100 against each parasite antigen by modified agglutination tests to determine whether antibodies to either parasite were present in the blood. Six of 62 samples (10%) were positive for T. gondii, with 2 samples having titers of 1:25 and 4 having titers of 1:50. Four of 62 samples (6%) were positive for S. neurona, with 2 samples having titers of 1:25 and 2 having titers of 1:50.

  7. Prevalence of agglutinating antibodies to Toxoplasma gondii and Sarcocystis neurona in beavers (Castor canadensis) from Massachusetts

    USGS Publications Warehouse

    Jordan, C.N.; Kaur, T.; Koenen, K.; DeStefano, S.; Zajac, A.M.; Lindsay, D.S.

    2005-01-01

    The present study examined the seroprevalence of Toxoplasma gondii and Sarcocystls neurona in a population of beavers (Castor canadensis) from Massachusetts. Sixty-two blood samples were collected during the field seasons over 3 consecutive years from different animals. Blood was collected onto filter paper and shipped to the Department of Biomedical Sciences, Virginia Tech, Blacksburg, Virginia, for parasite testing. The samples were tested at dilutions of 1:25, 1:50, and 1:100 against each parasite antigen by modified agglutination tests to determine whether antibodies to either parasite were present in the blood. Six of 62 samples (10%) were positive for T. gondii, with 2 samples having titers of 1:25 and 4 having titers of 1:50. Four of 62 samples (6%) were positive for S. neurona, with 2 samples having titers of 1:25 and 2 having titers of 1:50. ?? American Society of Pathologists 2005.

  8. Quick serological detection of a cancer biomarker with an agglutinated supramolecular glycoprobe.

    PubMed

    He, Xiao-Peng; Hu, Xi-Le; Jin, Hong-Ying; Gan, Jiemin; Zhu, Huili; Li, Jia; Long, Yi-Tao; Tian, He

    2015-09-01

    While serology represents the forefront technique for cancer diagnosis, current clinical methods for the detection of serum biomarkers have flaws in terms of the need of complicated manipulations, long analytical time, and high cost. Here, we develop a supramolecular glycoprobe for the quick serological detection of a cancer biomarker. The probe formed by agglutination between self-assembled glyco-gold nanoparticles and a lectin shows subtle optical variations upon the competitive recognition of a glycoprotein biomarker secreted by cancer cells, tumor-bearing mice, as well as clinical cancer patients, with no response to a series of controls including the serum of hepatitis patients. This research provides an insight into the development of effective tools for serological diagnosis of cancer.

  9. Studying red blood cell agglutination by measuring electrical and mechanical properties with a double optical tweezers

    NASA Astrophysics Data System (ADS)

    Fontes, Adriana; Fernandes, Heloise P.; de Thomaz, André A.; Barbosa, Luiz C.; Barjas-Castro, Maria L.; Cesar, Carlos L.

    2007-07-01

    The red blood cell (RBC) viscoelastic membrane contains proteins and glycolproteins embedded in, or attached, to a fluid lipid bilayer and are negatively charged, which creates a repulsive electric (zeta) potential between the cells and prevents their aggregation in the blood stream. The basis of the immunohematologic tests is the interaction between antigens and antibodies that causes hemagglutination. The identification of antibodies and antigens is of fundamental importance for the transfusional routine. This agglutination is induced by decreasing the zeta-potential through the introduction of artificial potential substances. This report proposes the use of the optical tweezers to measure the membrane viscosity, the cell adhesion, the zeta-potential and the size of the double layer of charges (CLC) formed around the cell in an electrolytic solution. The adhesion was quantified by slowly displacing two RBCs apart until the disagglutination. The CLC was measured using the force on the bead attached to a single RBC in response to an applied voltage. The zeta-potential was obtained by measuring the terminal velocity after releasing the RBC from the optical trap at the last applied voltage. For the membrane viscosity experiment, we trapped a bead attached to RBCs and measured the force to slide one RBC over the other as a function of the relative velocity. After we tested the methodology, we performed measurements using antibody and potential substances. We observed that this experiment can provide information about cell agglutination that helps to improve the tests usually performed in blood banks. We also believe that this methodology can be applied for measurements of zeta-potentials in other kind of samples.

  10. Red blood cell membrane viscoelasticity, agglutination and zeta potential measurements with double optical tweezers

    NASA Astrophysics Data System (ADS)

    Fontes, Adriana; Fernandes, Heloise P.; Barjas-Castro, Maria L.; de Thomaz, André A.; de Ysasa Pozzo, Liliana; Barbosa, Luiz C.; Cesar, Carlos L.

    2006-02-01

    The red blood cell (RBC) viscoelastic membrane contains proteins and glycolproteins embedded in, or attached, to a fluid lipid bilayer and are negatively charged, which creates a repulsive electric (zeta) potential between the cells and prevents their aggregation in the blood stream. There are techniques, however, to decrease the zeta potential to allow cell agglutination which are the basis of most of the tests of antigen-antibody interactions in blood banks. This report shows the use of a double optical tweezers to measure RBC membrane viscosity, agglutination and zeta potential. In our technique one of the optical tweezers trap a silica bead that binds strongly to a RBC at the end of a RBCs rouleaux and, at the same time, acts as a pico-Newton force transducer, after calibration through its displacement from the equilibrium position. The other optical tweezers trap the RBC at the other end. To measure the membrane viscosity the optical force is measured as a function of the velocity between the RBCs. To measure the adhesion the tweezers are slowly displaced apart until the RBCs disagglutination happens. The RBC zeta potential is measured in two complimentary ways, by the force on the silica bead attached to a single RBC in response to an applied electric field, and the conventional way, by the measurement of terminal velocity of the RBC after released from the optical trap. These two measurements provide information about the RBC charges and, also, electrolytic solution properties. We believe this can improve the methods of diagnosis in blood banks.

  11. 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.

  12. 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.

  13. 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.

  14. 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.

  15. 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.

  16. Agglutination of Histoplasma capsulatum by IgG Monoclonal Antibodies against Hsp60 Impacts Macrophage Effector Functions▿

    PubMed Central

    Guimarães, Allan Jefferson; Frases, Susana; Pontes, Bruno; de Cerqueira, Mariana Duarte; Rodrigues, Marcio L.; Viana, Nathan Bessa; Nimrichter, Leonardo; Nosanchuk, Joshua Daniel

    2011-01-01

    Histoplasma capsulatum can efficiently survive within macrophages, facilitating H. capsulatum translocation from the lung into the lymphatics and bloodstream. We have recently generated monoclonal antibodies (MAbs) to an H. capsulatum surface-expressed heat shock protein of 60 kDa (Hsp60) that modify disease in a murine histoplasmosis model. Interestingly, the MAbs induced different degrees of yeast cell agglutination in vitro. In the present study, we characterized the agglutination effects of the antibodies to Hsp60 on H. capsulatum yeast cells by light microscopy, flow cytometry, dynamic light scattering, measuring zeta potential, and using optical tweezers. We found that immunoglobulin Gs (IgGs) to Hsp60 cause H. capsulatum aggregation dependent on the (i) concentration of MAbs, (ii) MAb binding constant, and (iii) IgG subclass. Furthermore, infection of macrophages using agglutinates of various sizes after incubation with different Hsp60-binding MAbs induced association to macrophages through distinct cellular receptors and differentially affected macrophage antifungal functions. Hence, the capacity of IgG MAbs to agglutinate H. capsulatum significantly impacted pathogenic mechanisms of H. capsulatum during macrophage infection, and the effect was dependent on the antibody subclass and antigen epitope. PMID:21134968

  17. Detection of Salmonella enterica serovar Enteritidis (SE) Antibodies in Serum Using A Polystyrene Bead/SE Flagella Agglutination Assay

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Serologic screening of flocks can be an important method to detect Salmonella enteritidis (SE) infections but can be labor intensive or lack specificity. Our goal was to develop a rapid agglutination assay using SE flagella adsorbed to polystyrene beads as a simple, relatively specific test to dete...

  18. Rapid latex particle agglutination test for Escherichia coli strains of porcine origin producing heat-labile enterotoxin.

    PubMed Central

    Finkelstein, R A; Yang, Z S; Moseley, S L; Moon, H W

    1983-01-01

    A latex particle agglutination test previously shown to be suitable for the rapid identification of Escherichia coli strains of human origin producing heat-labile enterotoxin (R. A. Finkelstein and Z. Yang, J. Clin. Microbiol. 18:23-28) is equally applicable to strains of porcine origin. PMID:6361056

  19. 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.

  20. 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.

  1. 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.

  2. 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.

  3. 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.

  4. 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.

  5. 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.

  6. Usefulness of PCR and Antigen Latex Agglutination Test with Samples Obtained by Transthoracic Needle Aspiration for Diagnosis of Pneumococcal Pneumonia

    PubMed Central

    García, Amparo; Rosón, Beatriz; Pérez, José Luis; Verdaguer, Ricard; Dorca, Jordi; Carratalà, Jordi; Casanova, Aurora; Manresa, Frederic; Gudiol, Francesc

    1999-01-01

    In a large number of cases, the etiology of community-acquired pneumonia (CAP) is not established. Some cases are probably caused by Streptococcus pneumoniae. Transthoracic needle aspiration (TNA) culture has a limited sensitivity which might be improved by antigen detection or gene amplification techniques. We evaluated the capacity of a PCR assay and a latex agglutination test to detect S. pneumoniae in samples obtained by TNA from 95 patients with moderate-to-severe CAP. Latex agglutination and PCR had sensitivities of 52.2 and 91.3%, specificities of 88.7 and 83.3%, positive predictive values of 62.3 and 65.6%, and negative predictive values of 83.3 and 96.5%, respectively, when culture techniques were used as the “gold standard.” When we considered expanded criteria for the diagnosis of pneumococcal pneumonia as a standard for our calculations, latex agglutination and PCR had sensitivities of 53.6 and 89.7%, specificities of 93.0 and 90.0%, positive predictive values of 78.9 and 81.3%, and negative predictive values of 80.3 and 94.7%, respectively. The additional diagnosis provided by the PCR assay compared to latex agglutination was 12.2% (95% confidence interval of the difference from 0.4 to 20.1%). PCR was more sensitive than TNA culture, particularly in patients who had received prior antibiotic therapy (83.3 versus 33.3%). Although PCR is a very sensitive and specific technique, it has not proved to be cost-effective in clinical practice. Conversely, latex agglutination is a fast and simple method whose results might have significant implications for initial antibiotic therapy. PMID:9986837

  7. Congenital dyserythropoietic anaemia (CDA) with severe gout, rare Kell phenotype and erythrocyte, granulocyte and platelet membrane reduplication: a new variant of CDA type II.

    PubMed

    Lowenthal, R M; Marsden, K A; Dewar, C L; Thompson, G R

    1980-02-01

    A 43-year-old man with lifelong anaemia showed features which indicate him to have a previously undescribed variant of congenital dyserythropoietic anaemia (CDA), type II. The main clinical features--of which the first two are unique or very unusual in CDA--have been severe tophaceous gout, massive splenomegaly, gall stones mecessitating cholecystectomy and haemosiderosis affecting the liver and probably the heart. At age 41 he sustained a spontaneous retinal detachment. In the peripheral blood there were large numbers of nucleated red blood cells and marked macrocytosis; otherwise the picture was typical of CDA type II. The bone marrow contained many bi- and multi-nucleated erythrocyte precursors. There were increased levels of a number of red cell enzymes and a slightly raised level of HbF. Uncharacteristically, the red cells failed to lyse with acidified normal serum. The cells were strongly agglutinated by anti-i and were of the rare Kpb-negative phenotype. Plasma lipid analysis showed very low levels of cholesterol and vitamin E. Lipid peroxidation was markedly increased. Ultrastructural studies showed reduplication of the erythrocyte, granulocyte, and platelet cell membranes.

  8. 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.

  9. 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.

  10. 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.

  11. 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.

  12. 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

  13. 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

  14. 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.

  15. HEREDITARY OSTEOPETROSIS OF THE RABBIT

    PubMed Central

    Pearce, Louise; Brown, Wade H.

    1948-01-01

    The manifestations and course of an hereditary disease of the rabbit are reported. The condition is present at birth and is invariably fatal, generally in the 4th and 5th weeks of age. Retardation and eventual cessation of growth with marked reduction in size are conspicuous characteristic symptoms. The condition, which first occurred in the backcross progeny of a pure bred Dutch male rabbit, is inherited. It is determined by the expression of a simple recessive unit factor, affected individuals being homozygous for the factor. Rabbits heterozygous for the factor are identified only by appropriate breeding tests. The condition is not sex-linked. The disease has a remarkable resemblance to osteopetrosis or marble bone disease of infants and children with respect to signs and general course and also, as may be stated in anticipation of later discussions (5, 6), to the characteristic abnormal condition of the skeleton. PMID:18103397

  16. European Rabbits as Reservoir for Coxiella burnetii

    PubMed Central

    González-Barrio, David; Maio, Elisa; Vieira-Pinto, Madalena

    2015-01-01

    We studied the role of European rabbits (Oryctolagus cuniculus) as a reservoir for Coxiella burnetii in the Iberian region. High individual and population seroprevalences observed in wild and farmed rabbits, evidence of systemic infections, and vaginal shedding support the reservoir role of the European rabbit for C. burnetii. PMID:25988670

  17. 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.

  18. 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.

  19. 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

  20. 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.

  1. 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.

  2. Evidence that forskolin binds to the glucose transporter of human erythrocytes

    SciTech Connect

    Lavis, V.R.; Lee, D.P.; Shenolikar, S.

    1987-10-25

    Binding of (4-/sup 3/H)cytochalasin B and (12-/sup 3/H)forskolin to human erythrocyte membranes was measured by a centrifugation method. Glucose-displaceable binding of cytochalasin B was saturable, with KD = 0.11 microM, and maximum binding approximately 550 pmol/mg of protein. Forskolin inhibited the glucose-displaceable binding of cytochalasin B in an apparently competitive manner, with K1 = 3 microM. Glucose-displaceable binding of (12-/sup 3/H)forskolin was also saturable, with KD = 2.6 microM and maximum binding approximately equal to 400 pmol/mg of protein. The following compounds inhibited binding of (12-/sup 3/H)forskolin and (4-/sup 3/H)cytochalasin B equivalently, with relative potencies parallel to their reported affinities for the glucose transport system: cytochalasins A and D, dihydrocytochalasin B, L-rhamnose, L-glucose, D-galactose, D-mannose, D-glucose, 2-deoxy-D-glucose, 3-O-methyl-D-glucose, phloretin, and phlorizin. A water-soluble derivative of forskolin, 7-hemisuccinyl-7-desacetylforskolin, displaced equivalent amounts of (4-/sup 3/H)cytochalasin B or (12-/sup 3/H)forskolin. Rabbit erythrocyte membranes, which are deficient in glucose transporter, did not bind either (4-/sup 3/H)cytochalasin B or (12-/sup 3/H)forskolin in a glucose-displaceable manner. These results indicate that forskolin, in concentrations routinely employed for stimulation of adenylate cyclase, binds to the glucose transporter. Endogenous ligands with similar specificities could be important modulators of cellular metabolism.

  3. A comparison of titers of anti-Brucella antibodies of naturally infected and healthy vaccinated cattle by standard tube agglutination test, microtiter plate agglutination test, indirect hemagglutination assay, and indirect enzyme-linked immunosorbent assay

    PubMed Central

    Mohan, Anju; Saxena, Hari Mohan; Malhotra, Puneet

    2016-01-01

    Aim: We determined the antibody response in cattle naturally infected with brucellosis and normal healthy adult cattle vaccinated during calf hood with strain 19. Materials and Methods: The antibody titers were measured by standard tube agglutination test (STAT), microtiter plate agglutination test (MAT), indirect hemagglutination assay (IHA), and indirect enzyme-linked immunosorbent assay (iELISA) as per standard protocols. Results: The mean STAT titers were 1.963±0.345 in infected cattle and 1.200±0.155 in healthy vaccinated cattle. The difference was extremely significant (p<0.0001). The mean MAT titers were 2.244±0.727 in infected cattle and 1.200±0.155 in healthy vaccinated cattle. The difference was very significant (p<0.005). The mean IHA titers in infected cattle were 2.284±0.574, and those in healthy vaccinated cattle were 1.200±0.155. The difference was extremely significant (p=0.0002). However, the difference in mean iELISA titers of infected cattle (1.3678±0.014) and healthy vaccinated cattle (1.367±0.014) was non-significant. The infected animals showed very high titers of agglutinating antibodies compared to the vaccinated animals. However, it cannot be ascertained whether these antibodies are due to vaccine or response to infection. Since the infected animals had been vaccinated earlier, the current infection may suggest that vaccination was unable to induce protective levels of antibody. The heightened antibody response after infection may also indicate a secondary immune response to the antigens common to the vaccine strain and wild Brucella organisms. Conclusion: The brucellosis infected animals showed very high titers of agglutinating antibodies compared to the vaccinated animals. PMID:27536032

  4. Clinico-pathological findings of Clostridium perfringens type D enterotoxaemia in goats and its hemolytic activity in different erythrocytes

    PubMed Central

    Ali Nasir, A.; Younus, M.; Rashid, A.; Abdul Khaliq, S.; Khan, E.; Shah, S. H.; Aslam, A.; Ghumman, M. A.; Joiya, M. H.

    2015-01-01

    The present investigation was conducted to study the effects of experimental Clostridium perfringens type D enterotoxaemia in teddy goats. Clinical signs started to appear after 30 min of experimental infection like anorexia, diarrhea, dehydration, frothing and dyspnea. Gross lesions consisted of severe congestion in tissues of varying intensity with enlarged mesenteric lymph nodes while histological examination revealed edema of lungs, kidney, and lymph nodes and to some extent in brain along with hemorrhages in lungs and intestines. Clostridium perfringens type D carrying alpha and epsilon toxin genes were amplified with amplicon size about 247 bp and 665 bp, respectively. Human erythrocytes showed the highest hemolysis, 68%, followed by mice, 57%, against culture supernatants. The percentage of hemolysis was significantly higher at 37°C as compared to 25°C except for rabbit and dog. PMID:27175159

  5. Evaluation of Pyloriset Dry, a new rapid agglutination test for Helicobacter pylori antibody detection.

    PubMed Central

    Lozniewski, A; De Korwin, J D; Conroy, M C; Plenat, F; Weber, M

    1996-01-01

    We evaluated the performance of a new latex agglutination test, Pyloriset Dry (Orion Diagnostica, Espoo, Finland), in the simultaneous detection of immunoglobulin G (IgG), IgA, and IgM antibodies to Helicobacter pylori and compared it with that of the Pyloristat test (BioWhittaker, Fontenay-sous-Bois, France), an enzyme-linked immunosorbent assay detecting IgG to H. pylori, for 96 untreated dyspeptic patients who had undergone gastroduodenal endoscopy. Infection was diagnosed in 56 cases by positive culture and/or positive Giemsa stain and rapid urease test (antral biopsies) and was associated with chronic gastritis in 52 patients. Forty noninfected patients did not have chronic gastritis. The sensitivity of Pyloriset Dry was 91.1%. The sensitivity of Pyloristat was 91.1 or 82.1%, depending on whether equivocal results were considered positive or negative, respectively. Both tests had a specificity of 87.5%. Their performances were not statistically different. Thus, Pyloriset Dry is an alternative to serological tests for adults, particularly when a small number of serum samples has to be tested. PMID:8784587

  6. Quantitative capillary reversed passive latex agglutination test for C-reactive protein (CRP) in the dog.

    PubMed

    Tagata, K; Yokoyama, S; Ginbo, T; Honda, M; Okimura, T; Odakura, M; Nomura, M; Yamamoto, S

    1996-01-01

    A capillary reversed passive latex agglutination test (capillary RPLA) was developed which allows quantification of serum C-reactive protein (CRP) within approximately 15 min. The logarithmic regression line (calibration curve) obtained after measuring each CRP concentration three times in twofold dilutions of a standard canine serum containing 222 micrograms/ml of CRP was y = 6.394 + 0.030x (r = 0.995). Capillary RPLA permitted quantification of CRP in the range 6.9-222 micrograms/ml. The coefficients of variation ranged from 10.28% to 12.40%. The recovery rates (percentage recovery) of CRP by capillary RPLA were within the range 87% to 106%. On measuring the CRP concentrations in sera from 78 dogs by capillary RPLA, single radial immunodiffusion (SRID) and enzyme-linked immunosorbent assay (ELISA), close correlations were demonstrated between SRID and capillary RPLA (y = 7.250 + 1.109x, r = 0.978), between SRID and ELISA (y = 3.042 + 1.059x, r = 0.967), and between capillary RPLA and ELISA (y = 1.778 + 0.929x, r = 0.962). Capillary RPLA may be considered useful as a routine biochemical technique for measurement of serum CRP concentration in the dog.

  7. Cytotoxic, cell agglutinating, and syncytium forming effect of purified lectins from Ricinus communis on cultured cells.

    PubMed

    Koga, M; Ohtsu, M; Funatsu, G

    1979-10-01

    The toxicity of lectins from castor bean (Ricinus communis L.), ricin-D, ricin-E, and castor bean hemagglutinin, was investigated on five cultured cell lines. The differential effect of their constituent polypeptide chains was also investigated using these cell lines. Ricin-D, ricin-E, and castor bean hemagglutinin (CBH) possessed cytoagglutinating activity and cytotoxic activity to all five cell lines. These lectins showed the strongest toxicity to L5178Y cells, which are leukemic cells. The toxic activity of ricin-D was stronger than that of CBH in all cell lines. The constituent polypebtide chains of ricin-D and CBH were separated by DEAE-cellulose chromatography and designated as isoleucine chain and alanine chain denoted by their N-terminal amino acids. Only alanine chain of ricin-D was toxic to cells grown in vitro, whereas isoleucine chain of ricin-D and alanine chain of CBH were not toxic to the cells. Moreover, it was found that both lectins caused syncytium formation in NIH3T3 cells infected with Moloney leukemia virus and this cell fusion activity was shown to be exclusively associated with the alanine chain. Cytotoxic, cell agglutinating, and syncytium forming effect of the lectins is due to binding of the alanine chain of ricin-D to galactose-like residues of the membrane constituents of these cells.

  8. Interaction of bacterial endotoxine (lipopolysaccharide) with latex particles: application to latex agglutination immunoassays.

    PubMed

    Peula-García, J M; Molina-Bolivar, J A; Velasco, J; Rojas, A; Galisteo-González, F

    2002-01-15

    The latex agglutination immunoassay technique uses polymer colloids as carriers for antibodies or antigens to enhance the immunological reaction. In this work, the interaction of a lipopolysaccharide (LPS) of Brucella Melitensis with two conventional latexes has been studied. Some experiments on the physical adsorption of the LPS onto these polystyrene beads have been performed and several complexes with different coverage degrees were obtained by modifying the incubation conditions. Regarding the application in the development of diagnostic test systems, it is advisable to study the latex-LPS complexes from an electrokinetic and colloidal stability point of view. The complexes were electrokinetically characterized by measuring the electrophoretic mobility under different redispersion conditions. The colloidal stability was determined by simple turbidity measurements. Experimental and theoretical data have been employed to study the molecular disposition of the LPS in the latex particle surface to compare with the outer membrane of bacterial cells. Latex complexes covered by different LPS amounts showed high colloidal stability and adequate immunoreactivity that remains for a long time period.

  9. A systematic review on the microscopic agglutination test seroepidemiology of bovine leptospirosis in Latin America.

    PubMed

    Pinto, Priscila da Silva; Libonati, Hugo; Penna, Bruno; Lilenbaum, Walter

    2016-02-01

    The diagnosis of leptospirosis commonly relies on serology, which has three issues that are referred: the sampling, the antigen panel, and the cutoff point. We propose a systematic review of the bovine leptospirosis in Latin America, in order to provide a better understanding of the evolution of the research and of the seroepidemiology of bovine leptospirosis in that region. Internet databases were consulted over the year of 2014. Inclusion criteria for analysis included serosurvey using microscopic agglutination test (MAT), a relevant number of animals, the presence in the antigen panel of at least one representant of serogroup Sejroe, and a cutoff point of ≥100. A total of 242 articles that referred to cattle, leptospir*, and one region of Latin America was found. Only 105 articles regarding to serosurveys using MAT were found in several countries, and 61 (58.1 %) met all the inclusion criteria. In conclusion, this systematic review demonstrated a high prevalence of the infection (75.0 % at herd level and 44.2 % at animal level), with predominance of strains of serogroup Sejroe (80.3 %). It was evident that there is the necessity of more studies in several countries, as well as the need for greater standardization in studies, especially with regard to the adopted cutoff point at serological tests.

  10. Reading development in agglutinative languages: evidence from beginning, intermediate, and adult Basque readers.

    PubMed

    Acha, Joana; Laka, Itziar; Perea, Manuel

    2010-04-01

    Do typological properties of language, such as agglutination (i.e., the morphological process of adding affixes to the lexeme of a word), have an impact on the development of visual word recognition? To answer this question, we carried out an experiment in which beginning, intermediate, and adult Basque readers (n=32 each, average age=7, 11, and 22 years, respectively) needed to read correctly versus incorrectly inflected words embedded in sentences. Half of the targets contained high-frequency stems, and the other half contained low-frequency stems. To each stem, four inflections of different lengths were attached (-a, -ari, -aren, and -arentzat, i.e., inflectional sequences). To test whether the process of word recognition was modulated by the knowledge of word structure in the language, half of the participants' native language was Basque and the other half's native language was Spanish. Children showed robust effects of frequency and length of inflection that diminished with age. In addition, the effect of length of inflection was modulated by the frequency of the stem and by the native language. Taken together, these results suggest that word recognition develops from a decoding strategy to a direct lexical access strategy and that this process is modulated by children's knowledge of the inflectional structure of words from the beginning of their reading experience.

  11. Type 2B von Willebrand disease associated with the release of platelet agglutinates from megakaryocytes in the bone marrow.

    PubMed

    Slayton, William B; Patel, Milin; Sola-Visner, Martha; Harris, Neil; Rivers, Angela; Montgomery, Robert R; Friedman, Kenneth D

    2008-09-01

    We report a child with thrombocytopenia since birth, circulating platelet agglutinates, and a tendency to bleed. A bone marrow aspirate revealed large platelet clumps within the bone marrow and megakaryocyte nuclei surrounded by halos of clumped platelets. Laboratory evaluation revealed type 2B von Willebrand disease. Gene sequencing revealed a G to C mutation at base 3923 of the VWF gene. This mutation was previously described in a family with circulating platelet clumps and abnormal megakaryopoiesis with release of clumped platelets in culture. This same mutation was previously described in a family with circulating platelet aggregates and abnormalities of platelet release from megakaryocytes in vitro. Presence of megakaryocytes with halos of clumped platelets in our patient suggests that platelet agglutinate occurs in the bone marrow in some type 2B von Willebrand disease patients.

  12. 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.

  13. 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.

  14. 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.

  15. [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.

  16. 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.

  17. 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

  18. Organization and Expression of Plasmodial Genes Required for Erythrocyte Invasion

    DTIC Science & Technology

    1989-12-22

    S 0ED GROUP Su-GROUP Malaria, Vaccine , Molecular biology, Recombinant DNA, .06 13 Merozoite, Erythrocyte,.4-. 3 19 AIS T CT (Caft"Wu an Mt,.," of...j. Ravetch, J.V., Young, J. and Poste, G. (1985) Molecular genetic strategies for the development of anti-malarial vaccines . Biotechnology 3, 729. k...Perkins, M. and Pavetch, J.V. (1985) Interaction of P. falciparum merozoite proteins with the erythrocyte surface. In Vaccines , Cold Spring Harbor

  19. 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

  20. Comparison of optomagnetic and AC susceptibility readouts in a magnetic nanoparticle agglutination assay for detection of C-reactive protein.

    PubMed

    Fock, Jeppe; Parmvi, Mattias; Strömberg, Mattias; Svedlindh, Peter; Donolato, Marco; Hansen, Mikkel Fougt

    2017-02-15

    There is an increasing need to develop biosensor methods that are highly sensitive and that can be combined with low-cost consumables. The use of magnetic nanoparticles (MNPs) is attractive because their detection is compatible with low-cost disposables and because application of a magnetic field can be used to accelerate assay kinetics. We present the first study and comparison of the performance of magnetic susceptibility measurements and a newly proposed optomagnetic method. For the comparison we use the C-reactive protein (CRP) induced agglutination of identical samples of 100nm MNPs conjugated with CRP antibodies. Both methods detect agglutination as a shift to lower frequencies in measurements of the dynamics in response to an applied oscillating magnetic field. The magnetic susceptibility method probes the magnetic response whereas the optomagnetic technique probes the modulation of laser light transmitted through the sample. The two techniques provided highly correlated results upon agglutination when they measure the decrease of the signal from the individual MNPs (turn-off detection strategy), whereas the techniques provided different results, strongly depending on the read-out frequency, when detecting the signal due to MNP agglomerates (turn-on detection strategy). These observations are considered to be caused by differences in the volume-dependence of the magnetic and optical signals from agglomerates. The highest signal from agglomerates was found in the optomagnetic signal at low frequencies.

  1. Evaluation of a C-reactive protein latex agglutination detection test with sera from patients with sexually transmitted diseases.

    PubMed Central

    Schalla, W O; Arko, R J; Thompson, S E

    1984-01-01

    A total of 149 sera, including 79 pre- and posttreatment sera from 33 patients with disseminated gonococcal infections, 18 from patients with uncomplicated gonococcal infections, 6 from patients with pelvic inflammatory disease, 4 from patients with genital Chlamydia trachomatis infections, and 42 from normal volunteers, were examined for C-reactive protein with a latex agglutination C-reactive protein detection kit (Difco Laboratories, Detroit, Mich.). Results were quantitated with LC-Partigen C-reactive protein radial immuno-diffusion plates (Calbiochem-Behring, La Jolla, Calif.). Positive latex agglutination results were observed in all of the pretreatment sera and some of the posttreatment sera of patients with disseminated gonococcal infections and in two sera from patients with pelvic inflammatory disease, which corresponded to quantitative C-reactive protein levels in the radial immunodiffusion plates. C-reactive protein levels were not detectable in the serum samples from normal volunteers or patients with uncomplicated gonococcal infections or genital chlamydial infections. Positive latex agglutination occurred as early as 20 s in sera with high C-reactive protein levels, and all positive results were observed within 90 s of the 3-min test limit. Positive latex test results were obtained with C-reactive protein levels as low as 1 mg/dl (10 micrograms/ml). PMID:6440907

  2. 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

  3. Screening of agglutinins in marine algae from Fujian coast of China

    NASA Astrophysics Data System (ADS)

    Zheng, Yi; Lu, Hai-Sheng

    2002-09-01

    Thirty-three species of marine algae belonging to Rhodophyta, Phaeophyta and Chlorophyta from the Fujian coast were examined for agglutinins with different animal and human erythrocytes. Protein extracts from 26 species were active against at least one type of the erythrocytes tested. There were 3 species ( Grateloupia imbricata, Ishige foliacea and Entermorpha prolifera) whose extracts could agglutimate all the erythrocytes used. The lowest protein concentration required to produce erythrocyte agglutination varied remarkably, from 3.1 μg/ml to 500 μg/ml. The strongest activity was found in the agglutination of rabbit erythrocytes by Gloiopeltis furcata extract. Inhibition assays performed with nine mono- and bisaccharides indicated that agglutinations of rabbit erythrocytes by extracts of 7 species were inhibited by one or more types of the sugars assayed. The agglutinating activity shown by extracts of most species was not affected when the test solution was heated to 90°C, but was lost at 95°C 100°C. A few extracts lost their activity at 60°C, 65°C and 75°C, respectively.

  4. Microbiological quality of rabbit meat.

    PubMed

    Rodríguez-Calleja, Jose M; Santos, Jesús A; Otero, Andrés; García-López, María-Luisa

    2004-05-01

    World rabbit meat production is estimated to be over 1 million tons, and Spain is the third largest producer. Although rabbit meat is marketed and consumed worldwide, information on microbiological quality is very scarce. Here, we report indicator organisms, spoilage flora, sensory quality, and some physicochemical traits of 24 h postmortem chilled rabbit carcasses and prepackaged rabbit meat stored chilled in air for 0 to 3 days at the retail level. The mean total bacterial count (4.01 +/- 0.48 log CFU/g) for carcasses dressed at a small abattoir by a manual process was significantly lower (P < 0.05) than that (4.96 +/- 0.90 log CFU/g) for carcasses dressed at a large abattoir in automated slaughter lines. Both groups of carcasses had mean pH values of 5.98. The dominant contaminants on carcasses from the small abattoir were Pseudomonas, lactic acid bacteria, and yeasts. These microorganisms and Brochothrix thermosphacta were dominant on carcasses from the large abattoir. On prepacked hind legs (pH 6.26 +/- 0.18) stored at -1 to +1 degree C (supermarket 1), mean aerobic mesophilic count was 5.87 +/- 1.03 log CFU/g, and the major microbial groups were Pseudomonas, yeasts, lactic acid bacteria, and B. thermosphacta. On prepacked whole carcasses (pH 6.37 +/- 0.18) displayed at -1 to +5 degrees C (supermarket 2), mean aerobic mesophilic count was 6.60 +/- 1.18 and the same microbial groups were dominant. Relative Escherichia coli incidence was supermarket 2 > large abattoir > supermarket 1 > small abattoir. Overall, low numbers of coliforms, Enterobacteriaceae, psychrotrophic clostridia, coagulase-positive staphylococci, and molds were found. Sensory scores, pH values, and L-lactic acid content differentiated fresh carcasses from retail samples. Data obtained suggest that the microflora of chilled rabbit meat are different from those found on the meat of other animals.

  5. 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

  6. Taxonomic Notes on the Abyssal Agglutinated Benthic Foraminifera of the HEBBLE (High Energy Benthic Boundary Layer Experiment) Area (Lower Nova Scotian Continental Rise).

    DTIC Science & Technology

    1983-09-01

    Gulf of Mexico . Marine Sciences International, Woods Hole, Ma. Resig, J.M., 1981 Biogeography of benthic foraminifera of the northern Nazca plate and...HD-A133 743 TAXONOMIC NOTES ON THE ABYSSAL AGGLUTINATED BENTHIC t/i FORAMINIFERA OF-THE H..(U) WOODS HOLE OCEANOGRAPHIC INSTITUTION MRN M A KAMINSKI...Notes On The Abyssal Agglutinated Benthic Foraminifera September 1983 Of The HEBBLE Area (Lower Nova Scotian Continental Rise) 7. A41*heeW L Pinforv Mg

  7. 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

  8. 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.

  9. 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.

  10. 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

  11. 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.

  12. Comparison of agglutinating and neutralizing antibodies to serovar hardjo in sows immunized with two commercial whole culture polivalent anti-leptospira bacterins

    PubMed Central

    Soto, Francisco Rafael Martins; Pinheiro, Sônia Regina; Morais, Zenaide Maria; Gonçales, Amane Paldês; de Azevedo, Sérgio Santos; Bernardi, Fernanda; Camargo, Sebastião Rodrigues; Vasconcellos, Silvio Arruda

    2008-01-01

    It was performed the comparison of the intensity and duration of agglutinating and neutralizing antibodies to serovar Hardjo in swines vaccinated with two commercial anti-leptospira bacterins. Sows no reactive to 24 Leptospira sp serovars in the microscopic agglutination test (MAT) were divided in three groups: Group A (n=08): received two vaccine A doses with 30 days interval, Group B (n=08) two vaccine B doses with 30 days interval and Group C (n=08): control no vaccinated against leptospirosis.Blood samples were collected each 30 days during six months following the first vaccination. The sera were tested by MAT and growth inhibition test (GIT) to serovar Hardjo in order to evaluate respectively agglutinating and neutralizing antibodies. It was found that neutralizing antibodies persisted for a longer time than the agglutinating ones and that the absence of agglutinating antibodies does not means in the absence of the neutralizing. The peaks of agglutinating antibodies was obtained at least 30 days earlier than that produced by neutralizing. The duration of both kinds of antibodies measured differed between the two bacterines tested. The period for inducing neutralizing antibodies against serovar Hardjo indicated that gilts must be immunized with two doses of whole culture anti-leptospira bacterines applied 30 days each other at least 90 days before the first mating. For the maintenance of hight levels of neutralizing antibodies the revaccinations must be performed every six months after the first vaccination. PMID:24031250

  13. A critical review of published methods for analysis of red cell antigen-antibody reactions by flow cytometry, and approaches for resolving problems with red cell agglutination.

    PubMed

    Arndt, Patricia A; Garratty, George

    2010-07-01

    Flow cytometry operators often apply familiar white blood cell (WBC) methods when studying red blood cell (RBC) antigens and antibodies. Some WBC methods are not appropriate for RBCs, as the analysis of RBCs requires special considerations, for example, avoidance of agglutination. One hundred seventy-six published articles from 88 groups studying RBC interactions were reviewed. Three fourths of groups used at least one unnecessary WBC procedure for RBCs, and about one fourth did not use any method to prevent/disperse RBC agglutination. Flow cytometric studies were performed to determine the effect of RBC agglutination on results and compare different methods of preventing and/or dispersing agglutination. The presence of RBC agglutinates have been shown to be affected by the type of pipette tip used for mixing RBC suspensions, the number of antigen sites/RBC, the type and concentration of primary antibody, and the type of secondary antibody. For quantitation methods, for example, fetal maternal hemorrhage, the presence of agglutinates have been shown to adversely affect results (fewer fetal D+ RBCs detected).

  14. 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.

  15. The latex agglutination test versus counterimmunoelectrophoresis for rapid diagnosis of bacterial meningitis

    PubMed Central

    Bortolussi, Robert; Wort, Arthur J.; Casey, Stephanie

    1982-01-01

    A modified latex agglutination (LA) test was compared with Gram-staining and counterimmunoelectrophoresis (CIE) for the rapid detection in the cerebrospinal fluid (CSF) of antigen to Haemophilus influenzae type b, Neisseria meningitidis groups A, B and C, Escherichia coli K1, Streptococcus pneumoniae and group B streptococci, seven frequent causes of bacterial meningitis in children. Of 50 CSF samples from patients with culture-proven bacterial meningitis 90% were correctly shown by the LA test to contain antigen of the responsible organism. Gram-staining revealed organisms in 80% of 45 of these samples. In 75% of the 40 samples that were of sufficient volume for CIE, positive results for the appropriate antigen were obtained. The concentration of antigen detected in the CSF by the LA test varied from undetectable to 800 000 ng/ml. Patients with a high concentration (more than 2000 ng/ml or a positive result at dilutions of CSF over 1/8) were significantly more likely to have a poor response to therapy (two died and two had persistent pleocytosis or bacteria in the CSF) than patients with a lower concentration (4/16 v. 0/18, P < 0.05). After appropriate therapy was begun the concentration of antigen fell dramatically, but measurable amounts of antigen persisted in the CSF for up to 6 days. The LA test detected bacterial antigen at concentrations 2 to 70 times below the lower limit detected by CIE. In seven additional patients who had received antibiotics before lumbar puncture was performed the LA test detected antigen from meningitis-causing bacteria even though cultures of the CSF were sterile. In another 145 patients who did not have meningitis the results of the LA test were negative. The LA test, done as described in this article, is easier to perform than CIE and should be a useful addition to the diagnostic tests carried out on the CSF of any patient suspected of having meningitis. PMID:6749272

  16. Validation studies of the latex agglutination test for the detection of Trichinella larvae in meat products.

    PubMed

    Gayda, Jennifer; Reckinger, Sabine; Thaben, Nora; Nöckler, Karsten; Mayer-Scholl, Anne

    2016-11-15

    Human trichinellosis is a foodborne disease caused by ingestion of meat infected with Trichinella muscle larvae. To control Trichinella spp. infection in the European Union, all slaughtered pigs from holdings that are not officially recognized as applying controlled housing conditions and other animals susceptible to Trichinella infection and intended for human consumption should be examined by one of the approved digestion methods described in Regulation (EU) No. 2015/1375. In the past, Trichinella outbreaks due to the consumption of cured wild boar or pork products have been described in several European countries, making the identification of the larvae from these products relevant for Trichinella control. Therefore, this study aimed to validate the newly approved latex agglutination test (Trichin-L) for routine testing of cured meat products. The test was validated based on the OIE Guidelines using pork products spiked with Trichinella larvae. The sensitivity of the method varied greatly depending on the investigated meat product and was usually lower than for the gold standard, the magnetic stirrer method. The detection rate reached 80% for three larvae and 60% for one larva in cured pork sausages. A detection rate of 100% for three larvae and 50% for one larva was found in bacon. For frozen samples (-20°C) the Trichin-L kit is similarly sensitive as for cured samples. Further, to determine the performance of the test under field conditions, pork products from regions with known high Trichinella prevalences confiscated by customs authorities at two German international airports were analyzed. Problems associated with the Trichin-L test were incomplete digestion due to fatty ingredients, spices and very dry meat products, resulting in data which could not be evaluated. Therefore, the test is currently not suitable for the detection of Trichinella larvae in cured meat products and needs further adaptation steps to increase both usability and sensitivity.

  17. 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.

  18. 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

  19. 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.

  20. 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.

  1. Analysis of free zone electrophoresis of fixed erythrocytes performed in microgravity

    NASA Technical Reports Server (NTRS)

    Snyder, Robert S.; Rhodes, Percy H.; Herren, Blair J.; Miller, Teresa Y.; Seaman, Geoffrey V. F.

    1985-01-01

    A free fluid zone electrophoresis experiment was performed in the microgravity environment of Space Shuttle flight STS-3 (March 1983). The experiment was designed to confirm observations made on the Apollo-Soyuz mission of 1975 and to test the effect of high red blood cell (RBC) concentration on free fluid electrophoresis. Photographic documentation of cell zone progression in one-hour separations of mixtures of formaldehyde-fixed human and rabbit erythrocytes, which were subjected to a field of approximately 13 V/cm in low ionic strength buffer, was analyzed. One of two columns contained 2 x 10 to the 8th RBC/ml; (low concentration), and the other contained 1 x 10 to the 9th RBC/ml (high concentration). The observed and calculated leading edge displacements of the RBC in the two columns were in agreement, indicating the absence of unexpected effects of the reduced gravity environment. Post-flight analyses of the contents of the columns was not possible, and additional microgravity experiments are needed to evaluate the role of particle-particle interactions in concentrated suspensions undergoing electrophoresis.

  2. Immunosuppression abrogates resistance of young rabbits to Rabbit Haemorrhagic Disease (RHD).

    PubMed

    Marques, Raquel M; Teixeira, Luzia; Aguas, Artur P; Ribeiro, Joana C; Costa-e-Silva, António; Ferreira, Paula G

    2014-02-04

    Rabbit Haemorrhagic Disease (RHD) is caused by a calicivirus (RHDV) that kills 90% of infected adult European rabbits within 3 days. Remarkably, young rabbits are resistant to RHD. We induced immunosuppression in young rabbits by treatment with methylprednisolone acetate (MPA) and challenged the animals with RHDV by intramuscular injection. All of these young rabbits died within 3 days of infection due to fulminant hepatitis, presenting a large number of RHDV-positive dead or apoptotic hepatocytes, and a significant seric increase in cytokines, features that are similar to those of naïve adult rabbits infected by RHDV. We conclude that MPA-induced immunosuppression abrogates the resistance of young rabbits to RHD, indicating that there are differences in the innate immune system between young and adult rabbits that contribute to their distinct resistance/susceptibility to RHDV infection.

  3. Efficacy of a norvancomycin-loaded, PDLLA-coated plate in preventing early infection of rabbit tibia fracture.

    PubMed

    Fei, Jun; Yu, Hong-jun; Pan, Chang-jiang; Zhao, Chang-hong; Zhou, Yong-gang; Wang, Yan

    2010-05-12

    Despite improvements in surgical techniques and implant designs in orthopedic surgery, implantation-associated infections are still a challenging problem for surgeons. The goal of this study was to evaluate the efficacy of a norvancomycin-loaded, PDLLA-coated stainless steel plate vs an uncoated stainless steel plate in a rabbit model (n=50). The norvancomycin was delivered from a biodegradable poly(D,L-lactide) (PDLLA) coating of a stainless steel plate. Intraoperatively, rabbit tibia fractures were contaminated with Staphylococcus aureus (10(5) colony forming units) after plate implantation. The implants were either uncoated or coated with PDLLA and norvancomycin. In vivo drug release profiles showed that the norvancomycin release rate was decreased by increasing the time. The norvancomycin concentration in the tissue around the plate was higher than the minimum inhibitory concentration on the 14th day after implantation surgery. The animals were followed up for 28 days. Radiographic examinations were performed, and C-reactive protein and erythrocyte sedimentation rate were determined. Infection was evaluated by histological, microbiological, and radiological analysis. Eight of 25 rabbits (32%) implanted with the norvancomycin-loaded, PDLLA-coated plates were infected. Twenty-three of 25 rabbits (92%) implanted with the uncoated plates were infected (P<.05). The norvancomycin-loaded, PDLLA-coated plate may be used to treat open fractures to reduce the incidence of early infection.

  4. 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.

  5. The Rabbit Corneal Pocket Assay.

    PubMed

    Morbidelli, Lucia; Ziche, Marina

    2016-01-01

    The rabbit corneal micropocket angiogenesis assay uses the avascular cornea as a substrate canvas to study angiogenesis in vivo. Through the use of standardized slow-release pellets, a predictable angiogenic response is generated over the course of 1-2 weeks and then quantified. Uniform slow-release pellets are prepared by mixing purified angiogenic growth factors such as basic fibroblast growth factor or vascular endothelial growth factor and a synthetic polymer to allow slow release. A micropocket is surgically created in the rabbit cornea under anesthesia and a pellet implanted. On the days later, the angiogenic response is measured and qualified using a slit lamp, as well as the concomitant vascular phenotype or inflammatory features. The results of the assay are used to assess the ability of potential therapeutic molecules to modulate angiogenesis in vivo, both when released locally or given by ocular formulations or through systemic treatment. In this chapter, the experimental details of the rabbit cornea assay and technical implementations to the original protocol are described.

  6. 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.

  7. 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.

  8. 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.

  9. 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.

  10. 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

  11. 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.

  12. 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.

  13. 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.

  14. 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

  15. 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

  16. The Life Cycle of Entzia, an Agglutinated Foraminifer from the Salt Marshes in Transylvania

    NASA Astrophysics Data System (ADS)

    Kaminski, Michael; Telespan, Andreea; Balc, Ramona; Filipescu, Sorin; Varga, Ildiko; Görög, Agnes

    2013-04-01

    The small salt marshes associated with Miocene salt domes in Transylvania are host to a variety of marine organisms, including communities of halophytic plants as well as an agglutinated foraminifer that is normally found in coastal salt marshes worldwide. Originally described as the species Entzia tetrastoma by Daday (1884), the foraminifer is more widely known by the name Jadammina macrescens (Brady, 1870). Because the genus name Entzia has priority over Jadammina, the valid name of this taxon is Entzia macrescens (Brady, 1870). In 2007, we discovered a living population of Entzia inhabiting a small salt marsh just outside the town of Turda in central Transylvania, only a kilometer from the famous Maria Theresa Salt Mine. This is the first discovery of a living population of Entzia in Transylvania since the species was originally described in 1884. To determine whether or not the specimens we found represent a breeding population, samples were collected from the marsh on a monthly basis over the span of a year. This species can be found among the roots of the halophytic plants, in the uppermost one or two centimeters of the mud. Sediment samples were preserved in Vodka with Rose Bengal to distinguish living and dead specimens, and examined quantitatively. To document the life cycle of the species the following metrics were carried out: test size, abundance, number of chambers, ratio between live and dead specimens, and the diameter of the proloculus. An increase in the mean diameter of specimens was found from October to December. However the mean diameter decreased again in January, which suggests that asexual reproduction had apparently taken place. Small specimens again appeared in March, when sexual reproduction is presumed to have taken place. The median proloculus diameter was smallest in April and May, but the monthly changes in mean proloculus size within the population over the span of a year are not significant. However, specimens with largest

  17. 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.

  18. 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.

  19. 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.

  20. 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.

  1. 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.

  2. Optical Assay of Erythrocyte Function in Banked Blood

    PubMed Central

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

    2014-01-01

    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. PMID:25189281

  3. 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.

  4. 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.

  5. 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.

  6. 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.

  7. 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.

  8. 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.

  9. 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

  10. 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.

  11. 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.

  12. Eimeria stiedae: experimental infection in rabbits and the effect of treatment with toltrazuril and ivermectin.

    PubMed

    Cam, Yücel; Atasever, Ayhan; Eraslan, Gökhan; Kibar, Murat; Atalay, Oznur; Beyaz, Latife; Inci, Abdullah; Liman, Bilal Cem

    2008-05-01

    The aim of this study was to investigate the clinical, haematological, biochemical, lipid peroxidation, ultrasonographic and pathologic findings in hepatic coccidiosis induced by Eimeria stiedae in rabbits, and also to compare the treatment effects of both toltrazuril and ivermectin separately and in combination. In this study, 56 rabbits were divided into eight groups. The first group was designated as healthy control group. Rabbits were infected with 40.000 sporulated oocysts of E. stiedae. Groups 2, 3, 4, 5, 6, 7 and 8 were allocated as the infected control group, infected+toltrazuril-treated group, infected+ivermectin-treated group, infected+toltrazuril+ivermectin-treated group, non-infected+toltrazuril-treated group, non-infected+ivermectin-treated group, non-infected+toltrazuril+ivermectin-treated group, respectively. Haematocrit, Haemoglobin and MCV values as well as percentage of lymphocyte decreased in Groups 2 and 4 whereas leucocyte counts and percentage of granulocyte leucocyte increased. Serum GGT, ALT and AST activities increased but albumin value decreased. Plasma MDA concentrations increased whereas erythrocyte CAT, GSH-Px, and SOD activities decreased. Mean oocyst numbers in per gram faeces (epg values) increased in both groups during the study. Ultrasonographic examination revealed that the liver was enlarged and had hyperechogenic parenchyma. Bile ducts were dilated and hyperechogenic and the gall bladder was dilated. The livers of these animals were enlarged and typical macroscopic and microscopic findings of coccidiosis were present. Treatment with toltrazuril and toltrazuril+ivermectin combination were highly effective in reducing faecal oocyst output in infected rabbits. Haematological, biochemical and lipid peroxidation parameters and, ultrasonographic findings of the liver were close to control values for Groups 3 and 5. Necropsy of these animals showed no visible lesions related to hepatic coccidiosis although a few oocysts were detected

  13. Development of a peptide based latex agglutination assay for serotype identification of foot and mouth disease virus.

    PubMed

    Kaur, Dilpreet; Kaur, Gurpreet; Chandra, Mudit; Saxena, Hari M; Dwivedi, Padam N

    2013-02-01

    Out of 200 serum samples collected from cattle (142) and buffaloes (58) of various ages and sexand subjected to latex agglutination test (LAT) using serotype specific peptides (O, A, Asia 1) and also with peptide for non-structural protein 2B (NSP-2B), 114 (70%) samples were positive against FMDV type 'O', 102 (51%) against serotype 'A' and 104 (52%) against serotype 'Asia 1'. With NSP-2B peptide a total of 71 (35.5%) samples were positive. The results suggest that LAT could be used for the diagnosis of foot and mouth disease virus as it is easy, cheap and effective test.

  14. A sugar-functionalized amphiphilic pillar[5]arene: synthesis, self-assembly in water, and application in bacterial cell agglutination.

    PubMed

    Yu, Guocan; Ma, Yingjie; Han, Chengyou; Yao, Yong; Tang, Guping; Mao, Zhengwei; Gao, Changyou; Huang, Feihe

    2013-07-17

    A novel sugar-functionalized amphiphilic pillar[5]arene containing galactose groups as the hydrophlic part and alkyl chains as the hydrophobic part was designed and synthesized. It self-assembles in water to produce nanotubes as confirmed by TEM, SEM, and fluorescence microscopy. These nanotubes, showing low toxicity to both cancer and normal cells, can be utilized as excellent cell glues to agglutinate E. coli. The existence of galactoses on these nanotubes provides multivalent ligands that have high affinity for carbohydrate receptors on E. coli.

  15. 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

  16. Detection of hemagglutinins in cultures of squirrel monkey intestinal trichomonads.

    PubMed Central

    Pindak, F F; Gardner, W A; Mora de Pindak, M; Abee, C R

    1987-01-01

    Intestinal trichomonads are very common inhabitants of captive squirrel monkeys. In evaluating the potential pathogenicity of these organisms, we encountered hitherto unknown hemagglutinins in their culture fluids. The cytopathic effect associated with a number of the isolates resembled that caused by vacuolating viruses. We have ruled out conventional viruses as the cause of the cytopathic effect and as the source of the hemagglutinin. The agglutinin has some of the basic characteristics of lectins. Parallel experiments demonstrated agglutination of erythrocytes from squirrel monkeys, humans, dogs, cats, guinea pigs, and horses, with the first two types being the most sensitive. Relatively less agglutination was seen with rat erythrocytes. Agglutination of sheep, rabbit, chicken, and bovine erythrocytes was virtually absent. Images PMID:3571468

  17. 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

  18. 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).

  19. 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.

  20. 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.

  1. 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.

  2. 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.

  3. 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.

  4. 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.

  5. 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.

  6. 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.

  7. 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.

  8. IMMUNIZATION OF RABBITS TO INFECTIOUS PAPILLOMATOSIS

    PubMed Central

    Shope, Richard E.

    1937-01-01

    Two intraperitoneal injections of either infectious or non-infectious rabbit papilloma suspensions actively immunize rabbits against papillomatosis. The capacity of the non-infectious suspensions to immunize is considered as evidence that they contain papilloma virus even though none can be demonstrated by the usual infection test. PMID:19870596

  9. Mapping superficial lymphatic territories in the rabbit.

    PubMed

    Soto-Miranda, Miguel A; Suami, Hiroo; Chang, David W

    2013-06-01

    Little is known about the anatomy of the lymphatic system in the rabbit with regard to relationships between the lymphatic vessel and lymph node. According to our previous studies in human cadavers and canines, the superficial lymphatic system could be divided into lymphatic territories. The aim of this study was to completely map the superficial lymphatic system in the rabbit. We used our microinjection technique and histological analysis for dissecting studies and recently developed indocyanine green (ICG) fluorescent lymphography for demonstrating dynamic lymph flow in living rabbits. Real-time ICG fluorescent lymphography was performed in two living New Zealand White rabbits, and direct dye microinjection of the lymphatic vessels was performed in eight dead rabbits. To assess the relationships between the vascular and lymphatic systems in rabbits, we performed radiocontrast injection into arteries in two dead rabbits prior to the lymphatic injection. The ICG fluorescent lymphography revealed eight lymphatic territories in the preauricular, submandibular, root of the lateral neck, axillary, lumbar, inguinal, root of the tail, and popliteal regions. We injected blue acrylic dye into every lymphatic vessel 0.1 mm in diameter or larger. We then dissected and chased the stained lymphatic vessels proximally until the vessels connected to the first tier lymph node. This procedure was repeated throughout the body until all the relationships between the lymphatic vessels and lymph nodes were defined. The lymphatic system of the rabbit could be defined as eight lymphatic territories, each with its own lymphatic vessels and lymph node.

  10. 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.

  11. 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...

  12. 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...

  13. 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...

  14. 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...

  15. 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.

  16. [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.

  17. A prototype of the direct agglutination test kit (DAT-Canis) for the serological diagnosis of canine visceral leishmaniasis.

    PubMed

    Oliveira, Edward; Saliba, Juliana Wilke; Oliveira, Diana; Dias, Edelberto Santos; Paz, Gustavo Fontes

    2016-05-15

    This report describes the stege I/II development of a new direct agglutination test (DAT) for the diagnosis of canine visceral leishmaniasis (CVL) using freeze-dried antigen produced Coomassie blue-stained Leishmania (Leishmania) infantum promastigotes. In stage I, 16 canine serum samples, collected from eight dogs carrying CVL and eight healthy dogs, were assessed with the DAT using 2-mercaptoethanol (2-ME), N-acetyl-cysteine (NAC), kaolin or NAC plus urea (NAC+U) to improve the assay conditions. Stage II assessed the diagnostic accuracy with 100 serum samples collected from dogs with symptomatic CVL and clinically healthy dogs, comparing the four different sample diluents. The CVL-DAT prototype kit showed equivalent performances when 2-ME, NAC or NAC+U were used: 97.1% sensitivity (CI: 83-99.8%), 97% specificity (CI: 88.5-99.5%) and a 97% diagnostic accuracy (CI: 90.8-99.2). With kaolin, a 94.1% sensitivity (CI: 79-99%), 97% specificity (CI: 88.5-99.5%) and 96% diagnostic accuracy were observed (CI: 89.5-98.7), with no statistically significant differences among the four reagents (p=1.0). The NAC plus urea in sample diluent decreased non-specific agglutination, promoted a better defined sharp-edged blue spot and was thus chosen as a component for the new DAT prototype to diagnose canine VL, designated DAT-Canis.

  18. Insights into the Antimicrobial Mechanism of Action of Human RNase6: Structural Determinants for Bacterial Cell Agglutination and Membrane Permeation

    PubMed Central

    Pulido, David; Arranz-Trullén, Javier; Prats-Ejarque, Guillem; Velázquez, Diego; Torrent, Marc; Moussaoui, Mohammed; Boix, Ester

    2016-01-01

    Human Ribonuclease 6 is a secreted protein belonging to the ribonuclease A (RNaseA) superfamily, a vertebrate specific family suggested to arise with an ancestral host defense role. Tissue distribution analysis revealed its expression in innate cell types, showing abundance in monocytes and neutrophils. Recent evidence of induction of the protein expression by bacterial infection suggested an antipathogen function in vivo. In our laboratory, the antimicrobial properties of the protein have been evaluated against Gram-negative and Gram-positive species and its mechanism of action was characterized using a membrane model. Interestingly, our results indicate that RNase6, as previously reported for RNase3, is able to specifically agglutinate Gram-negative bacteria as a main trait of its antimicrobial activity. Moreover, a side by side comparative analysis with the RN6(1–45) derived peptide highlights that the antimicrobial activity is mostly retained at the protein N-terminus. Further work by site directed mutagenesis and structural analysis has identified two residues involved in the protein antimicrobial action (Trp1 and Ile13) that are essential for the cell agglutination properties. This is the first structure-functional characterization of RNase6 antimicrobial properties, supporting its contribution to the infection focus clearance. PMID:27089320

  19. [Methicillin resistance detection in Staphylococcus aureus: comparison between conventional methods and MRSA-Screen latex agglutination technique].

    PubMed

    Soloaga, R; Corso, A; Gagetti, P; Faccone, D; Galas, M

    2004-01-01

    Methicillin-resistant Staphylococcus aureus (MRSA) is a significant pathogen that has emerged over the last four decades, causing both nosocomial and community-acquired infections. Rapid and accurate detection of methicillin resistance in S. aureus is important for the use of appropriate antimicrobial therapy and for the control of nosocomial spread of MRSA strains. We evaluated the efficiency of conventional methods for detection of methicillin resistance such as the disk diffusion, agar dilution, oxacillin agar screen test, and the latex agglutination test MRSA-Screen latex, in 100 isolates of S. aureus, 79 mecA positive and 21 mecA negative. The MRSA-Screen latex (Denka Seiken, Niigata, Japón), is a latex agglutination method that detects the presence of PLP-2a, product of mecA gene in S. aureus. The PCR of the mecA gene was used as the "gold standard" for the evaluation of the different methods tested. The percentages of sensitivity and specificity were as follows: disk difusión 97 and 100%, agar dilution 97 and 95%, oxacillin agar screen test 100 and 100%, and MRSA-Screen latex, 100 and 100 %. All methods presented high sensitivity and specificity, but MRSA-Screen latex had the advantage of giving a reliable result, equivalent to PCR, in only 15 minutes.

  20. Prevalence of agglutinating antibodies to Sarcocystis neurona in skunks (Mephitis Mephitis), raccoons (Procyon lotor), and opossums (Didelphis Virginiana) from Connecticut.

    PubMed

    Mitchell, Sheila M; Richardson, Dennis J; Cheadle, M Andy; Zajac, Anne M; Lindsay, David S

    2002-10-01

    Equine protozoal myeloencephalitis is the most important protozoan disease of horses in North America and is usually caused by Sarcocystis neurona. Natural cases of encephalitis caused by S. neurona have been reported in skunks (Mephitis mephitis) and raccoons (Procyon lotor). Opossums (Didelphis spp.) are the only known definitive host. Sera from 24 striped skunks, 12 raccoons, and 7 opossums (D. virginiana) from Connecticut were examined for agglutinating antibodies to S. neurona using the S. neurona agglutination test (SAT) employing formalin-fixed merozoites as antigen. The SAT was validated for skunk sera using pre- and postinfection serum samples from 2 experimentally infected skunks. Of the 24 (46%) skunks 11 were positive, and all 12 raccoons were positive for S. neurona antibodies. None of the 7 opossums was positive for antibodies to S. neurona. These results suggest that exposure to sporocysts of S. neurona by intermediate hosts is high in Connecticut. The absence of antibodies in opossums collected from the same areas is most likely because of the absence of systemic infection in the definitive host.

  1. [AGGLUTINATION OF MESOPHYLL PLASTIDS AND OBLITERATION OF PHLOEM SIEVE TUBES ARE THE TOTAL RESULT OF SEASONAL PAUSES IN PHOTOSYNTHATE EXPORT].

    PubMed

    Gamalei, Yu V

    2015-01-01

    Chloroplast agglutination and sieve tube obliteration are related to the different plant tissues: the agglutination--to the leaf mesophyll, and the obliteration--to the axis phloem. Being equally produced by photosynthate export dynamics, both phenomena are synchronous and can be used for diagnostics of seasonal flashes and pauses of photosynthetic activity with equal success. The nature of the mobility of chloroplast and their shuttle displacements from the nuclear envelope to the cell periphery connected with export dynamics have been established. It is assumed that nuclear envelope is the base structure of the endoplasmic reticulum (ER) inside which the chloroplasts are localized. Activation of photosynthesis and sugar accumulation inside the ER induces its expansion followed by centrifugal diffusion of chloroplasts. Come back effect--ER collapse, its return to the source--can be induced by the blockade of photosynthesis. Centripetal collapse is accompanied by plastid concentration around the nuclear envelope. Displacements of ER and the chloroplasts dislocating inside it are reversible. It depends on seasonal fluctuations of photosynthesis and export intensities. Changes in the volume of sieve tubes, which are due to the same reason, are irreversible. Each seasonal wave of photosynthesis and sugar export forms new series of sieve tubes, replacing obliterated ones.

  2. siRNA delivery targeting to the lung via agglutination-induced accumulation and clearance of cationic tetraamino fullerene

    NASA Astrophysics Data System (ADS)

    Minami, Kosuke; Okamoto, Koji; Doi, Kent; Harano, Koji; Noiri, Eisei; Nakamura, Eiichi

    2014-05-01

    The efficient treatment of lung diseases requires lung-selective delivery of agents to the lung. However, lung-selective delivery is difficult because the accumulation of micrometer-sized carriers in the lung often induces inflammation and embolization-related toxicity. Here we demonstrate a lung-selective delivery system of small interfering RNA (siRNA) by controlling the size of carrier vehicle in blood vessels. The carrier is made of tetra(piperazino)fullerene epoxide (TPFE), a water-soluble cationic tetraamino fullerene. TPFE and siRNA form sub-micrometer-sized complexes in buffered solution and these complexes agglutinate further with plasma proteins in the bloodstream to form micrometer-sized particles. The agglutinate rapidly clogs the lung capillaries, releases the siRNA into lung cells to silence expression of target genes, and is then cleared rapidly from the lung after siRNA delivery. We applied our delivery system to an animal model of sepsis, indicating the potential of TPFE-based siRNA delivery for clinical applications.

  3. Thermal cataract formation in rabbits

    SciTech Connect

    Kramar, P.; Harris, C.; Guy, A.W.

    1987-01-01

    Intraocularly circulating hot water was used to produce cataracts in nine eyes of seven rabbits by maintaining their retrolental temperatures between 43 degrees C and 45 degrees C. A rapid rate of heating (1.3 degrees C/min) plus a sharp temperature gradient across the eye may have been contributing factors in the consistent production of cataracts at these temperatures. Biomicroscopy and light microscopy showed lens changes similar to those associated with acute exposure to microwave radiation. These findings support the assumption that microwave cataractogenesis is due to the local production of elevated temperatures.

  4. Novel Calicivirus Identified in Rabbits, Michigan, USA

    PubMed Central

    Wise, Annabel G.; Bolin, Steven R.; Mullaney, Thomas P.; Kiupel, Matti; Maes, Roger K.

    2009-01-01

    We report a disease outbreak in a Michigan rabbitry of a rabbit calicivirus distinct from the foreign animal disease agent, rabbit hemorrhagic disease virus (RHDV). The novel virus has been designated Michigan rabbit calicivirus (MRCV). Caliciviruses of the Lagovirus genus other than RHDV have not been described in US rabbit populations. The case-fatality rate was 32.5% (65/200). Clinical signs included hemorrhage and sudden death, with hepatic necrosis. Analysis of viral RNA sequence from >95% of the viral genome showed an average similarity of 79% with RHDV. Similarity of the predicted MRCV capsid amino acid sequence ranged from 89.8% to 91.3%, much lower than the 98% amino acid similarity between RHDV strains. Experimentally infected rabbits lacked clinical disease, but MRCV was detected in tissues by PCR. We propose that MRCV primarily causes subclinical infection but may induce overt RHD-like disease under certain field conditions. PMID:19961675

  5. In vitro effects of 50 Hz magnetic fields on oxidatively damaged rabbit red blood cells.

    PubMed

    Fiorani, M; Biagiarelli, B; Vetrano, F; Guidi, G; Dachà, M; Stocchi, V

    1997-01-01

    The aim of this study was to investigate the effects of 50 Hz magnetic fields (0.2-0.5 mT) on rabbit red blood cells (RBCs) that were exposed simultaneously to the action of an oxygen radical-generating system, Fe(II)/ascorbate. Previous data obtained in our laboratory showed at the exposure of rabbit erythrocytes or reticulocytes to Fe(II)/ascorbate hexokinase inactivation, whereas the other glycolytic enzymes do not show any decay. We also observed depletion of reduced glutathione (GSH) content with a concomitant intracellular and extracellular increase in oxidized glutathione (GSSG) and a decrease in energy charge. In this work we investigated whether 50 Hz magnetic fields could influence the intracellular impairments that occur when erythrocytes or reticulocytes are exposed to this oxidant system, namely, inactivation of hexokinase activity, GSH depletion, a change in energy charge, and hemoglobin oxidation. The results obtained indicate the a 0.5 mT magnetic field had no effect on intact RBCs, whereas it increased the damage with Fe(II)/ascorbate to a 0.5 mT magnetic field induced a significant further decay in hexokinase activity (about 20%) as well as a twofold increase in methemoglobin production compared with RBCs that were exposed to the oxidant system alone. Although further studies will be needed to determine the physiological implications of these data, the results reported in this study demonstrate that the effects of the magnetic fields investigated are able to potentiate the cellular damage induced in vitro by oxidizing agents.

  6. 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...

  7. 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

  8. 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

  9. 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.

  10. 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.

  11. 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...

  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...

  13. 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...

  14. 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.

  15. 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.

  16. Signatures of deterministic chaos in dyslipidemic erythrocytes under shear stress.

    PubMed

    Korol, A M; Rasia, R

    2003-03-01

    The erythrocytes deformation under shear stress, the viscoelastic properties and the effects observed on healthy donors and on dyslipidemic patients are investigated from the point of view of nonlinear dynamics. Finally it is suggested that the different results obtained could be useful with regard to pathophysiological disturbances and their treatment.

  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. 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.

  19. 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.

  20. 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.

  1. 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.

  2. Associations of erythrocyte fatty acid patterns with insulin resistance

    Technology Transfer Automated Retrieval System (TEKTRAN)

    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...

  3. 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

  4. Behavioral fever in newborn rabbits

    NASA Technical Reports Server (NTRS)

    Satinoff, E.; Mcewen, G. N., Jr.; Williams, B. A.

    1976-01-01

    New Zealand white rabbit pups aged 12 to 72 hr were divided into three groups and given an intraperitoneal injection of Pseudomonas polysaccharide, a saline vehicle alone, and no treatment, respectively. The animals injected with pyrogen and maintained at an ambient temperature of 32 C for 2 hr did not develop fever. When placed in a thermally graded alleyway, the animals injected with pyrogen selected gradient positions that represented significantly higher temperatures than controls injected with saline. Further stay at selected positions for 5 min caused a considerable increase in the rectal temperature of the pyrogen-injected pups but not that of controls. The results support the hypothesis that newborn rabbits will develop a fever by behavioral means after a single injection of an exogenous pyrogen if the opportunity for thermoregulatory behavior is present. No fever develops if the pups must rely solely on internal thermoregulatory mechanisms. The behavioral system for producing a fever is mature at birth, but an adequate system of internal reflexes does not appear to develop for some days.

  5. [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.

  6. 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+}.

  7. 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.

  8. 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.

  9. Prozone effects in microscopic agglutination tests for leptospirosis in the sera of mice infected with the pathogenic Leptospira interrogans serovar Canicola

    PubMed Central

    Shimabukuro, Fabio Hiroto; da Costa, Veruska Maia; da Silva, Rodrigo Costa; Langoni, Hélio; da Silva, Aristeu Vieira; de Carvalho, Lídia Raquel; Domingues, Paulo Francisco

    2013-01-01

    Mice experimentally infected with a pathogenic strain of Leptospira interrogans serovar Canicola produced false negative results (prozone effect) in a microscopic agglutination test (MAT). This prozone effect occurred in several serum samples collected at different post-infection times, but it was more prominent in samples collected from seven-42 days post-infection and for 1:50 and 1:100 sample dilutions. This phenomenon was correlated with increased antibody titres in the early post-infection phase. While prozone effects are often observed in serological agglutination assays for the diagnosis of animal brucellosis and human syphilis, they are not widely reported in leptospirosis MATs. PMID:23903987

  10. 9 CFR 354.124 - Quarantine of diseased rabbits.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... 9 Animals and Animal Products 2 2012-01-01 2012-01-01 false Quarantine of diseased rabbits. 354... INSPECTION AND CERTIFICATION VOLUNTARY INSPECTION OF RABBITS AND EDIBLE PRODUCTS THEREOF Inspection Procedures; Ante-Mortem Inspections § 354.124 Quarantine of diseased rabbits. If live rabbits, which...

  11. 9 CFR 354.124 - Quarantine of diseased rabbits.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 9 Animals and Animal Products 2 2011-01-01 2011-01-01 false Quarantine of diseased rabbits. 354... INSPECTION AND CERTIFICATION VOLUNTARY INSPECTION OF RABBITS AND EDIBLE PRODUCTS THEREOF Inspection Procedures; Ante-Mortem Inspections § 354.124 Quarantine of diseased rabbits. If live rabbits, which...

  12. 9 CFR 354.124 - Quarantine of diseased rabbits.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... 9 Animals and Animal Products 2 2013-01-01 2013-01-01 false Quarantine of diseased rabbits. 354... INSPECTION AND CERTIFICATION VOLUNTARY INSPECTION OF RABBITS AND EDIBLE PRODUCTS THEREOF Inspection Procedures; Ante-Mortem Inspections § 354.124 Quarantine of diseased rabbits. If live rabbits, which...

  13. 9 CFR 354.124 - Quarantine of diseased rabbits.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... 9 Animals and Animal Products 2 2014-01-01 2014-01-01 false Quarantine of diseased rabbits. 354... INSPECTION AND CERTIFICATION VOLUNTARY INSPECTION OF RABBITS AND EDIBLE PRODUCTS THEREOF Inspection Procedures; Ante-Mortem Inspections § 354.124 Quarantine of diseased rabbits. If live rabbits, which...

  14. Factors affecting rabbit meat consumption among Spanish university students.

    PubMed

    González-Redondo, Pedro; Mena, Yolanda; Fernández-Cabanás, Victor M

    2010-01-01

    This study investigates the rabbit meat consumption patterns among students. Results of the 342 students surveyed at the University of Seville, Spain showed that more men than women consumed rabbit meat. More students consumed wild rabbit meat than farmed rabbit meat. The practice of hunting by students or their relatives, and the raising of rabbits for self-consumption, positively influenced the proportion of students that currently consumed rabbit meat. Most of the students that had eaten rabbit meat before but are currently non-eaters do so because of the meat's organoleptic characteristics or from lack of consumption habit.

  15. Encephalitozoon cuniculi in rabbits: Serological screening and histopathological findings.

    PubMed

    Maestrini, Giovanni; Ricci, Emanuele; Cantile, Carlo; Mannella, Riccardo; Mancianti, Francesca; Paci, Gisella; D'Ascenzi, Carlo; Perrucci, Stefania

    2017-02-01

    Serological prevalence of E. cuniculi infection was assessed in 183 rabbits from central Italy. In seropositive deceased rabbits, histopathological lesions were also evaluated. Sera from 118 rabbits from 6 intensive farms, 10 rabbits from 6 family farms, 16 rabbits from a zoo, 30 rabbits from 5 research laboratories and 9 pet rabbits from 9 different owners, were tested by an enzyme-linked immunosorbent assay. Data were statistically analysed. Tissue samples from brain and kidney of 10 deceased rabbits were formalin-fixed and subsequently analysed by histopathology and immunohistochemistry. Anti-E. cuniculi antibodies were found in 129/183 (70.5%) analysed sera. At statistical analysis, E. cuniculi seropositivity was significantly higher (p<0.05) in industrial and zoo rabbits. At histology, different degrees of pathological lesions were found in serological positive (9) deceased animals. In three rabbits deceased after showing neurological signs, the severity of the lesions was interpreted as a likely cause for their death.

  16. 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.

  17. 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.

  18. 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.

  19. Bobcat attack on a cottontail rabbit

    USGS Publications Warehouse

    Biggins, D.E.; Biggins, D.M.

    2006-01-01

    We observed an attack by a bobcat (Lynx rufus) on a cottontail rabbit (Sylvilagus) that involved stealthy approach by the cat for >1 h, followed by a 12.3-s chase covering 116.0 m for the cat and 128.4 m for the rabbit. During the chase, the route of the cat from starting point to kill site was more direct than the semi-circular route of the rabbit. Stride lengths for the cat and total distance covered by the chase were longer than those previously reported for bobcats.

  20. 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

  1. 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

  2. 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

  3. Metabolism of hydroanthracenones in rabbits

    PubMed Central

    Robertson, J. S.; Dunstan, P. J.

    1972-01-01

    1. The metabolism of 1-oxo-octahydro- and 2- and 9-oxoperhydro-anthracenes was investigated in rabbits. All compounds increased the urinary glucuronide content. 2. The 1-oxo and 2-oxo compounds were reduced to the corresponding alcohols whereas the 9-oxo compound was hydroxylated. 3. The reduction in vitro of these compounds and related ketones was investigated with three different enzyme systems (liver alcohol dehydrogenase, hydroxy steroid dehydrogenase, aromatic aldehyde–ketone reductase) in an attempt to explain the results in vivo. 4. Successful reduction of many ketones with aromatic aldehyde–ketone reductase suggests that the kidney may be of importance in the reduction in vivo of certain cyclic carbonyl compounds. PMID:5073736

  4. A new LDLa domain-containing C-type lectin with bacterial agglutinating and binding activity in amphioxus.

    PubMed

    Qu, Baozhen; Yang, Shuangshuang; Ma, Zengyu; Gao, Zhan; Zhang, Shicui

    2016-12-15

    Over 1200 C-type lectin gene models have been identified in amphioxus, but only a few of them have been functionally characterized. In this study, we identified a C-type lectin, BjCTL, with domain structure of LDLa-CTLD-EGF_Lam, the first such data in chordates. It was expressed mainly in the notochord and ovary in a tissue-dependent fashion. Recombinant BjCTL was characterized as a typical Ca(2+)-dependent carbohydrate-binding protein capable of agglutinating and binding to both Gram-negative and positive bacteria we tested. In addition, it specifically bound to insoluble lipopolysaccharide, lipoteichoic acid and peptidoglycan, which can be inhibited by galactose. We also showed that the interaction of BjCTL with the bacteria is primarily attributable to CTLD domain. Thus, BjCTL is a novel pattern recognition protein involved in lectin-mediated innate immunity.

  5. Gelatin particle indirect agglutination and enzyme-linked immunosorbent assay for diagnosis of strongyloidiasis using Strongyloides venezuelensis antigen.

    PubMed

    Huaman, Maria Cecilia; Sato, Yoshiya; Aguilar, Jose Luis; Terashima, Angelica; Guerra, Humberto; Gotuzzo, Eduardo; Kanbara, Hiroji

    2003-01-01

    Routine microscopical examination of stool specimens for diagnosis of strongyloidiasis is insensitive and serological methods using Strongyloides stercoralis antigen are at present not available for field studies. We evaluated 2 techniques, enzyme-linked immunosorbent assay (ELISA) and gelatin particle indirect agglutination (GPIA), using an antigen obtained from the rodent parasite, S. venezuelensis. Fifty-four Peruvian patients with different clinical forms of strongyloidiasis were studied: 12 asymptomatic, 31 symptomatic, and 11 hyperinfection cases. Our results demonstrate that both ELISA and GPIA using S. venezuelensis antigen are useful for diagnosis of strongyloidiasis, with sensitivities of 74.1% and 98.2%, respectively and a specificity of 100% for both techniques. We found that GPIA is a highly sensitive test for patients with suspected chronic infection and/or hyperinfection. In the hyperinfection cases, significantly lower concentrations of specific immunoglobulin antibodies and eosinophils (P < 0.001) were found compared with the asymptomatic and symptomatic cases.

  6. Rapid detection of infectious mononucleosis-associated heterophile antibodies by a novel immunochromatographic assay and a latex agglutination test.

    PubMed Central

    Farhat, S E; Finn, S; Chua, R; Smith, B; Simor, A E; George, P; Diena, B B; Diena, D; Skulnick, M

    1993-01-01

    A novel immunochromatographic assay, the CARDS O.S. MONO test (Pacific Biotech, San Diego, Calif.), and a latex agglutination test, the Infectious Mononucleosis Kit (Unipath Ltd., Hampshire, United Kingdom) were compared with the Paul-Bunnell-Davidsohn test. Of the 957 serum specimens studied, 78 were positive and 879 were negative by the Paul-Bunnell-Davidsohn test. After discrepancies were resolved by determining Epstein-Barr virus serology, the sensitivities of the CARDS O.S. MONO test and the Infectious Mononucleosis Kit were 91.0 and 96.2%, respectively, and both tests had a specificity and a positive predictive value of 100% and a negative predictive value and overall agreement of greater than 99%. The results show that both tests can accurately detect infectious mononucleosis-associated heterophile antibodies. PMID:8315001

  7. K/Ar dating of lunar soils. IV - Orange glass from 74220 and agglutinates from 14259 and 14163

    NASA Technical Reports Server (NTRS)

    Alexander, E. C., Jr.; Coscio, M. R., Jr.; Dragon, J. C.; Saito, K.

    1980-01-01

    Total fusion Ar-40 - A-39 analyses of orange glass from lunar soil 74220 combined with the sums of earlier stepwise heating data by other workers have yielded a precise K/Ar isochron with a slope corresponding to an age of 3.66 + or - 0.03 G.y. for the orange glass. The result is in marginal agreement with Huneke's (1978) age of 3.60 + or - 0.04 G.y. for 74220 glass. The Ar systematics in the agglutinates from 14259 and 14163 are dominated by volume correlated argon. Step-wise heating analyses yield data which define experimentally reproducible linear arrays in Ar-40/Ar-36 vs. K-40/Ar-36 diagrams. The slopes of these arrays correspond formally to very old ages, but it is not clear, however, that such ages have any physical significance.

  8. Effects of potassium channel toxins from Leiurus quinquestriatus hebraeus venom on responses to cromakalim in rabbit blood vessels.

    PubMed Central

    Strong, P. N.; Weir, S. W.; Beech, D. J.; Hiestand, P.; Kocher, H. P.

    1989-01-01

    1. The effects of fractionated Leiurus quinquestriatus hebraeus venom on cromakalim-induced 86Rb+ efflux in rabbit aortic smooth muscle were examined. 2. Crude venom (0.1-30 micrograms ml-1) produced a concentration-dependent decrease of 1 microM cromakalim-induced 86Rb+ response. The maximum blocking activity attainable was approximately 60%. 3. Fractionation of crude venom by gel permeation chromatography and subsequent chromatography on a cation ion-exchange column, produced two fractions (X and XI), active in the 86Rb+ blocking assay. 4. Fraction XII contained charybdotoxin (approximately 85% pure). After a final high performance liquid chromatography (h.p.l.c.) purification step, the purified toxin failed to inhibit the cromakalim-stimulated 86Rb+ efflux although it was a potent inhibitor of A23187-induced K+ flux in human erythrocytes and the large conductance calcium-activated potassium channel in rabbit portal vein smooth muscle. 5. Subsequent purification of fraction X by h.p.l.c. yielded a minor peak which contained 86Rb+ blocking activity. This subfraction was also capable of inhibiting apamin-sensitive, angiotensin II-stimulated K+ flux in guinea-pig hepatocytes. 6. It is concluded that the potassium channel opened by cromakalim in rabbit aortic smooth muscle is not blocked by charybdotoxin but by another distinct toxin in the venom of Leiurus quinquestriatus hebraeus. PMID:2531622

  9. Combined albumin and bicarbonate induces head-to-head sperm agglutination which physically prevents equine sperm-oviduct binding.

    PubMed

    Leemans, Bart; Gadella, Bart M; Stout, Tom A E; Sostaric, Edita; De Schauwer, Catharina; Nelis, Hilde; Hoogewijs, Maarten; Van Soom, Ann

    2016-04-01

    In many species, sperm binding to oviduct epithelium is believed to be an essential step in generating a highly fertile capacitated sperm population primed for fertilization. In several mammalian species, this interaction is based on carbohydrate-lectin recognition. D-galactose has previously been characterized as a key molecule that facilitates sperm-oviduct binding in the horse. We used oviduct explant and oviduct apical plasma membrane (APM) assays to investigate the effects of various carbohydrates; glycosaminoglycans; lectins; S-S reductants; and the capacitating factors albumin, Ca(2+) and HCO3(-) on sperm-oviduct binding in the horse. Carbohydrate-specific lectin staining indicated that N-acetylgalactosamine, N-acetylneuraminic acid (sialic acid) and D-mannose or D-glucose were the most abundant carbohydrates on equine oviduct epithelia, whereas D-galactose moieties were not detected. However, in a competitive binding assay, sperm-oviduct binding density was not influenced by any tested carbohydrates, glycosaminoglycans, lectins or D-penicillamine, nor did the glycosaminoglycans induce sperm tail-associated protein tyrosine phosphorylation. Furthermore, N-glycosidase F (PNGase) pretreatment of oviduct explants and APM did not alter sperm-oviduct binding density. By contrast, a combination of the sperm-capacitating factors albumin and HCO3(-) severely reduced (>10-fold) equine sperm-oviduct binding density by inducing rapid head-to-head agglutination, both of which events were independent of Ca(2+) and an elevated pH (7.9). Conversely, neither albumin and HCO3(-) nor any other capacitating factor could induce release of oviduct-bound sperm. In conclusion, a combination of albumin and HCO3(-) markedly induced sperm head-to-head agglutination which physically prevented stallion sperm to bind to oviduct epithelium.

  10. Evaluation of the MycoAKT latex agglutination test for rapid diagnosis of Mycobacterium avium complex infections.

    PubMed

    Olano, J P; Holmes, H; Woods, G L

    1998-01-01

    Rapid diagnosis of Mycobacterium avium complex (MAC) bacteremia is important for management of patients with the acquired immunodeficiency syndrome who have disseminated MAC. The purpose of this study was to determine the reliability of the MycoAKT latex agglutination test for direct detection of MAC in positive mycobacterial blood cultures. First, colonies of isolates of previously identified mycobacteria, including 35 MAC, were tested. Of the 55 isolates evaluated, 33 were identified as MAC by the latex test, including 31 of the known MAC and 2 M. chelonae (sensitivity, 88.6%; specificity, 90.0%). Second, broth from 20 ESP II and 20 BACTEC 12B bottles seeded with isolates of MAC were tested. Aliquots from 19 (95%) ESP II cultures and 16 (80%) 12B cultures were positive by the latex test. In phase 3, broth from 115 signal-positive ESP II blood cultures were tested by latex agglutination. Forty-three subcultures from these bottles grew mycobacteria (41 MAC and 2 Mycobacterium tuberculosis complex); the remainder grew no organisms. Broth from 40 of the blood cultures (39 that grew MAC and 1 from which no organisms were recovered) were latex positive; thus, the sensitivity, specificity, and positive and negative predictive values of the latex test for direct identification of MAC in ESP II blood cultures were 95.1, 98.6, 97.5, and 97.3%, respectively. The mean time to detection of MAC was 14.6 days (range, 6-34 days) with the direct latex test, compared with 18.3 days (range, 9-36 days) with subculture and probe (p < 0.05).

  11. Use of the protective antigen of Erysipelothrix rhusiopathiae in the enzyme-linked immunosorbent assay and latex agglutination.

    PubMed

    Sato, H; Yamazaki, Y; Tsuchiya, K; Aoyama, T; Akaba, N; Suzuki, T; Yokoyama, A; Saito, H; Maehara, N

    1998-09-01

    To establish a safe and convenient serodiagnostic method for swine erysipelas, a purified protective protein antigen of Erysipelothrix rhusiopathiae, which included a large amount of protective protein (64 kDa protein), was used for enzyme-linked immunosorbent assay (ELISA) and the latex agglutination (LA) test. In the ELISA, the antisera to four different serovars (1a, 2, 5 and 20) of E. rhusiopathiae exhibit a positive reaction, while antisera to other species of bacteria (Listeria monocytogenes, Staphylococcus aureus, Streptococcus suis, Rhodococcus equi and Corynebacterium pseudotuberculosis) exhibit a negative reaction. In the LA test, the antisera to three different serovars (1a, 2 and 5) of E. rhusiopathiae reacted with P64-sensitized latex beads, while the antiserum to serovar 20 (2553 strain) did not. Moreover, the antisera to other species of bacteria (Listeria monocytogenes, Staphylococcus aureus, Streptococcus suis, Rhodococcus equi and Corynebacterium pseudotuberculosis) did not in this test. Comparing the results of the growth agglutination (GA), ELISA and LA tests of 284 swine sera, there was a high degree of correlation among the results. The detection of anti-E. rhusiopathiae antibodies in the GA, ELISA and LA tests were compared using sera from pigs immunized with P64, alkaline extract (AE) and live-cell vaccine (LV). In all three tests, anti-E. rhusiopathiae antibodies could be detected 1 week after immunization. The serum antibody titre as determined by the LA test increased moderately, as did that by the GA test, while that determined by ELISA increased rapidly. These results suggested that ELISA could be used to monitor changes in anti-E. rhusiopathiae antibody titre and the LA test could be used in the screening test for swine erysipelas.

  12. Bayesian estimation of sensitivity and specificity of the modified agglutination test and bioassay for detection of Toxoplasma gondii in free-range chickens

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Toxoplasma gondii infects virtually all warm-blooded animals worldwide. Serological tests, including the modified agglutination test (MAT), are often used to determine exposure to the parasite. The MAT can be used for all hosts because it does not need species-specific reagents and has been shown to...

  13. Identification and characterization of anti-microbial peptides from rabbit vaginal fluid.

    PubMed

    Patgaonkar, Mandar; Aranha, Clara; Bhonde, Gauri; Reddy, K V R

    2011-02-15

    Antimicrobial peptides (AMPs) serve as a first line of host defense and represent an important, though poorly understood components of the innate immune system. The present study was an attempt to identify and characterize the major molecules having anti-bacterial activities from the vaginal fluid of rabbit, Oryctologus cuniculus. AMPs from the rabbit vaginal fluid (RVF) were identified in the acid extracts of pooled RVF samples after RP-HPLC purification. The protein, RVFAMP was effective against gram negative (Escherichia coli and Pseudomonas aeruginosa) and gram positive (Staphylococcus aureus and Streptococcus pyogenes) bacteria. The results of acid urea-PAGE-gel overlay assay (AU-PAGE-GOA) demonstrated clear zone of growth inhibition of E. coli corresponding to 6 and 15 kDa protein bands. LC-MS data of these proteins indicated that 15 kDa protein consists of lysozyme, lipopolysaccharide binding protein (LBP), hemoglobin-α and β subunits (Hb-α/β), whereas 9 kDa protein band consists of transthyretin and calcyclin while uteroglobulin and neutrophil antibacterial peptide-5 (NAMP-5) are present in the 6 kDa protein band. Of the eight proteins, Hb-α derived protein was further characterized, as it showed the highest Probability Based Mowse Score (PBMS) of 288. A 25mer peptide, RVFHbαp was active against several clinical pathogens as demonstrated by minimum inhibitory concentration (MIC) and radial diffusion assays (RDA). The interaction of RVFHbαP with bacterial liposome membrane was assessed by calcein dye leakage assay. RVFHbαP did not show cytotoxicity against human endocervical cells (End1/E6E7) or erythrocytes. RT-PCR and immunofluorescence results revealed the expression of RVFHbαP mRNA and protein in rabbit vaginal tissue. To the best our knowledge, this is the first report describing the detection of AMPs in RVFs. In conclusion, these studies indicated that vaginal epithelial cells (VEC) derived RVFHbαP may have therapeutic potential in the

  14. 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

  15. 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.

  16. [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.

  17. Pre-erythrocytic malaria vaccines: identifying the targets

    PubMed Central

    Duffy, Patrick E; Sahu, Tejram; Akue, Adovi; Milman, Neta; Anderson, Charles

    2013-01-01

    Pre-erythrocytic malaria vaccines target Plasmodium during its sporozoite and liver stages, and can prevent progression to blood-stage disease, which causes a million deaths each year. Whole organism sporozoite vaccines induce sterile immunity in animals and humans and guide subunit vaccine development. A recombinant protein-in-adjuvant pre-erythrocytic vaccine called RTS,S reduces clinical malaria without preventing infection in field studies and additional antigens may be required to achieve sterile immunity. Although few vaccine antigens have progressed to human testing, new insights into parasite biology, expression profiles and immunobiology have offered new targets for intervention. Future advances require human trials of additional antigens, as well as platforms to induce the durable antibody and cellular responses including CD8+ T cells that contribute to sterile protection. PMID:23176657

  18. Progress in Dodecafluoropentane Emulsion as a Neuroprotective Agent in a Rabbit Stroke Model

    PubMed Central

    Woods, S.D.; Skinner, R.D.; Ricca, A.M.; Brown, A.T.; Lowery, J.D.; Borrelli, M.J.; Lay, J.O.; Culp, W.C.

    2013-01-01

    Introduction Dodecafluoropentane emulsion (DDFPe) in 250 nm nanodroplets seems to swell modestly to accept and carry large amounts of oxygen in the body at >29°C. Small particle size allows oxygen delivery even into hypoxic tissue unreachable by erythrocytes. Using permanent cerebral embolic occlusion in rabbits, we assessed DDFPe dose response as a neuroprotectant at 7 and 24 hours post-embolization without lysis of arterial obstructions and investigated blood pharmacokinetics. Methods New Zealand White rabbits (N=56) received cerebral angiography and embolic spheres (diameter=700-900 μm) occluded middle and/or anterior cerebral arteries. Intravenous DDFPe dosing (2% w/v emulsion) began at 60 minutes and repeated every 90 minutes until sacrifice at 7 or 24 hours post-embolization. Seven hour groups: 1) Control (embolized without treatment, N=6), and DDFPe treatment: 2) 0.1ml/kg (N=7), 3) 0.3ml/kg (N=9), 4) 0.6ml/kg (N=8). Twenty-four hour groups: 5) Control (N=16), and DDFPe treatment: 6) 0.1ml/kg (N=10). Infarcts as percent of total brain volume were determined using vital stains on brain sections. Other alert normal rabbits (N=8) received IV doses followed by rapid arterial blood sampling and GC-MS analysis. Results Percent infarct volume means significantly decreased for all DDFPe treated groups compared with controls, p=<0.004 to <0.03. Blood DDFP (gas) half-life was 1.45±0.17 minutes with R=0.958. Mean blood clearance was 78.5±24.9 ml/min/kg (mean±SE). Conclusions Intravenous DDFPe decreases ischemic stroke infarct volumes. Blood half-life values are very short. The much longer therapeutic effect, >90 minutes, suggests multiple compartments. Lowest effective dose and maximum effective therapy duration are not yet defined. Rapid development is warranted. PMID:23813100

  19. 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.

  20. 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.

  1. 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.

  2. 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.

  3. 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.

  4. 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.

  5. 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

  6. 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

  7. 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.

  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 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.

  10. Blood antioxidant status and erythrocyte lipid peroxidation following distance running.

    PubMed

    Duthie, G G; Robertson, J D; Maughan, R J; Morrice, P C

    1990-10-01

    The relationship between prolonged exercise, oxidative stress, and the protective capacity of the antioxidant defense system has been determined. Venous blood samples were removed from seven trained athletes before and up to 120 h after completion of a half-marathon for measurements of blood antioxidants, antioxidant enzymes, and indices of lipid peroxidation. Plasma creatine kinase (CK) activity, an index of muscle damage, increased (P less than 0.05) to a maximum 24 h after the race but this was not accompanied by changes in conjugated dienes and thiobarbituric acid reactive substances (TBARS), which are indices of lipid peroxidation. An increase (P less than 0.05) in plasma cholesterol concentration (4%) immediately after the race was similar to the change in plasma volume (6%). However, transient increases (P less than 0.05) immediately postrace in the plasma concentrations of uric acid (24%), vitamin A (18%), and vitamin C (34%) were only partly accounted for by the fluid shifts. The immediate postrace increases in alpha- and gamma-tocopherol did not attain statistical significance. Erythrocyte antioxidant enzyme activities were unaffected by the exercise but the alpha- and gamma-tocopherol concentrations progressively increased (P less than 0.001 and P less than 0.05, respectively) up to 48 h postrace. Paradoxically, 24 h after the race erythrocyte susceptibility to in vitro peroxidation was markedly elevated (P less than 0.01). This enhanced susceptibility to peroxidation was maintained even at 120 h postrace and did not correspond to changes in the age of the red cell population. A decrease (P less than 0.001) in total erythrocyte glutathione immediately after the half-marathon was mainly due to a reduction in the reduced form (GSH). The results show that when trained athletes run a comparatively short distance sufficient to result in some degree of muscle damage but which is insufficient to cause elevations in plasma indices of lipid peroxidation

  11. Rabbit haemorrhagic disease: field epidemiology and the management of wild rabbit populations.

    PubMed

    Cooke, B D

    2002-08-01

    Rabbit haemorrhagic disease (RHD) has become established in wild rabbit populations throughout Western Europe, Australia and New Zealand. The abundance of wild rabbits has been significantly reduced, particularly in drier areas of southern Spain, inland Australia and South Island New Zealand. A detailed knowledge of the epidemiology of RHD is essential for the management of the disease in natural rabbit populations, either to rebuild or to control populations. When RHD first spread among naive wild rabbits, epidemiological studies provided unique information on the rate of spread, the possible role of insect vectors in transmission, and the correlation between the impact of disease on populations and climatic variables. Current research shows a consistent pattern of epidemiology between Europe and Australasia. Typically, the most severe epizootics of RHD occur among young sub-adult rabbits which have lost age-related resilience and maternal antibodies. However, the timing of these outbreaks reflects climatic variables that determine the breeding season of the rabbits and the periods when RHD virus (RDHV) is most likely to persist and spread. Further factors that may complicate epidemiology include the possibility that non-pathogenic RHDV-like viruses are present in natural rabbit populations. Additionally, the question of how the virus persists from year to year remains unresolved; persistence in carrier rabbits is a possibility. Understanding of the epidemiology of RHD is now sufficiently advanced to consider the possibility of manipulating rabbit populations to alter the epidemiological pattern of RHD and thereby maximise or minimise the mortality caused by the disease. Altering the epidemiology of RHD in this manner would assist the management of wild rabbit populations either for conservation or pest control purposes.

  12. Genomic Analysis of Companion Rabbit Staphylococcus aureus

    PubMed Central

    Holmes, Mark A.; Harrison, Ewan M.; Fisher, Elizabeth A.; Graham, Elizabeth M.; Parkhill, Julian; Foster, Geoffrey; Paterson, Gavin K.

    2016-01-01

    In addition to being an important human pathogen, Staphylococcus aureus is able to cause a variety of infections in numerous other host species. While the S. aureus strains causing infection in several of these hosts have been well characterised, this is not the case for companion rabbits (Oryctolagus cuniculus), where little data are available on S. aureus strains from this host. To address this deficiency we have performed antimicrobial susceptibility testing and genome sequencing on a collection of S. aureus isolates from companion rabbits. The findings show a diverse S. aureus population is able to cause infection in this host, and while antimicrobial resistance was uncommon, the isolates possess a range of known and putative virulence factors consistent with a diverse clinical presentation in companion rabbits including severe abscesses. We additionally show that companion rabbit isolates carry polymorphisms within dltB as described as underlying host-adaption of S. aureus to farmed rabbits. The availability of S. aureus genome sequences from companion rabbits provides an important aid to understanding the pathogenesis of disease in this host and in the clinical management and surveillance of these infections. PMID:26963381

  13. Erythrocyte deformability and aggregation in homozygous sickle cell disease.

    PubMed

    Vayá, Amparo; Collado, Susana; Dasí, Maria Angeles; Pérez, Maria Luz; Hernandez, Jose Luis; Barragán, Eva

    2014-01-01

    Rheological properties of homozygous sickle cell anaemia (SCA) show marked heterogeneity, which may be explained in part by the concomitance of alpha genotypes or beta haplotypes, along with hydroxurea (HU) treatment. To further clarify this issue, in 11 homozygous patients with SCA in the steady state and in 16 healthy controls, we analysed erythrocyte deformability (ED) in a Rheodyn SSD by means of the Elongation Index (EI) at 12, 30 and 60 Pa, and erythrocyte aggregation at stasis (EA0) and at 3 sec-1 (EA1) in a Myrenne aggregometer along with fibrinogen, biochemical and haematological parameters. When compared with controls, homozygous (SS) patients showed a lower EI at all the shear stresses tested (p < 0.01) and higher EA0 (p < 0.014), but not higher EA1 (p = 0.076). Fibrinogen did not show statistical differences (p = 0.642). In the Spearman's correlation IE60 correlated inversely with Hb S (p < 0.05) and directly with MCV, MCH and Hb F levels (p < 0.01). EA0 correlated inversely with MCV, MCH, Hb F (p < 0.01) and directly with Hb S (p < 0.05). HU treatment improved EI and EA0, but not EA1. This paradoxical behaviour of HU on erythrocyte aggregation merits further research to be clarified.

  14. Erythrocyte shape classification using integral-geometry-based methods.

    PubMed

    Gual-Arnau, X; Herold-García, S; Simó, A

    2015-07-01

    Erythrocyte shape deformations are related to different important illnesses. In this paper, we focus on one of the most important: the Sickle cell disease. This disease causes the hardening or polymerization of the hemoglobin that contains the erythrocytes. The study of this process using digital images of peripheral blood smears can offer useful results in the clinical diagnosis of these illnesses. In particular, it would be very valuable to find a rapid and reproducible automatic classification method to quantify the number of deformed cells and so gauge the severity of the illness. In this paper, we show the good results obtained in the automatic classification of erythrocytes in normal cells, sickle cells, and cells with other deformations, when we use a set of functions based on integral-geometry methods, an active contour-based segmentation method, and a k-NN classification algorithm. Blood specimens were obtained from patients with Sickle cell disease. Seventeen peripheral blood smears were obtained for the study, and 45 images of different fields were obtained. A specialist selected the cells to use, determining those cells which were normal, elongated, and with other deformations present in the images. A process of automatic classification, with cross-validation of errors with the proposed descriptors and with other two functions used in previous studies, was realized.

  15. Evidencing the Role of Erythrocytic Apoptosis in Malarial Anemia

    PubMed Central

    Totino, Paulo R. R.; Daniel-Ribeiro, Cláudio T.; Ferreira-da-Cruz, Maria de Fátima

    2016-01-01

    In the last decade it has become clear that, similarly to nucleated cells, enucleated red blood cells (RBCs) are susceptible to programmed apoptotic cell death. Erythrocytic apoptosis seems to play a role in physiological clearance of aged RBCs, but it may also be implicated in anemia of different etiological sources including drug therapy and infectious diseases. In malaria, severe anemia is a common complication leading to death of children and pregnant women living in malaria-endemic regions of Africa. The pathogenesis of malarial anemia is multifactorial and involves both ineffective production of RBCs by the bone marrow and premature elimination of non-parasitized RBCs, phenomena potentially associated with apoptosis. In the present overview, we discuss evidences associating erythrocytic apoptosis with the pathogenesis of severe malarial anemia, as well as with regulation of parasite clearance in malaria. Efforts to understand the role of erythrocytic apoptosis in malarial anemia can help to identify potential targets for therapeutic intervention based on apoptotic pathways and consequently, mitigate the harmful impact of malaria in global public health. PMID:28018860

  16. The free heme concentration in healthy human erythrocytes

    PubMed Central

    Aich, Anupam; Freundlich, Melissa; Vekilov, Peter G.

    2016-01-01

    Heme, the prosthetic group of hemoglobin, may be released from its host due to an intrinsic instability of hemoglobin and accumulate in the erythrocytes. Free heme is in the form of hematin (Fe3+ protoporphyrin IX OH) and follows several pathways of biochemical toxicity to tissues, cells, and organelles since it catalyzes the production of reactive oxygen species. To determine concentration of soluble free heme in human erythrocytes, we develop a new method. We lyse the red blood cells and isolate free heme from hemoglobin by dialysis. We use the heme to reconstitute horseradish peroxidase (HRP) from an excess of the apoenzyme and determine the HRP reaction rate from the evolution of the emitted luminescence. We find that in a population of five healthy adults the average free heme concentration in the erythrocytes is 21 ± 2 μM, ca. 100× higher than previously determined. Tests suggest that the lower previous value was due to the use of elevated concentrations of NaCl, which drive hematin precipitation and re-association with apoglobin. We show that the found hematin concentration is significantly higher than estimates based on equilibrium release and the known hematin dimerization. The factors that lead to enhanced heme release remain an open question. PMID:26460266

  17. Synthetic receptors for selectively detecting erythrocyte ABO subgroups.

    PubMed

    Seifner, Alexandra; Lieberzeit, Peter; Jungbauer, Christof; Dickert, Franz L

    2009-10-05

    Surface imprinting techniques with erythrocytes as templates yield polymer coatings with selective recognition sites towards red blood cells. The resulting cavities in the respective surface exhibit selectivity between blood subgroups as shown by Quartz Crystal Microbalance (QCM) measurements. Mass sensitive effects in the kilohertz range could be observed for concentrations down to 0.5 x 10(8) cells/mL. Frequency response as well as recovery of the sensor took place within a few minutes, indicating that no covalent binding is involved. Linear concentration dependence over a defined region provides ideal conditions for cross selectivity measurements. A1 imprinted sensor coatings resulted in an effect of 40 kHz when exposed to the template blood group, while A2 erythrocytes yielded just 11% of that value on the same layer. Furthermore, A2 imprinted coatings incorporated only one third the amount of A1 erythrocytes as compared to A2 ones. Therefore, imprinted materials depict the entire cell surface and utilize it for recognition, whereas natural antibodies bind on the defined antigen position and thus usually cannot distinguish between cells carrying different amounts of them.

  18. Transport of 3-bromopyruvate across the human erythrocyte membrane.

    PubMed

    Sadowska-Bartosz, Izabela; Soszyński, Mirosław; Ułaszewski, Stanisław; Ko, Young; Bartosz, Grzegorz

    2014-06-01

    3-Bromopyruvic acid (3-BP) is a promising anticancer compound because it is a strong inhibitor of glycolytic enzymes, especially glyceraldehyde 3-phosphate dehydrogenase. The Warburg effect means that malignant cells are much more dependent on glycolysis than normal cells. Potential complications of anticancer therapy with 3-BP are side effects due to its interaction with normal cells, especially erythrocytes. Transport into cells is critical for 3-BP to have intracellular effects. The aim of our study was the kinetic characterization of 3-BP transport into human erythrocytes. 3-BP uptake by erythrocytes was linear within the first 3 min and pH-dependent. The transport rate decreased with increasing pH in the range of 6.0-8.0. The Km and Vm values for 3-BP transport were 0.89 mM and 0.94 mmol/(l cells x min), respectively. The transport was inhibited competitively by pyruvate and significantly inhibited by DIDS, SITS, and 1-cyano-4-hydroxycinnamic acid. Flavonoids also inhibited 3-BP transport: the most potent inhibition was found for luteolin and quercetin.

  19. Metabolic homeostasis in the human erythrocyte: in silico analysis.

    PubMed

    de Atauri, Pedro; Ramírez, María José; Kuchel, Philip W; Carreras, José; Cascante, Marta

    2006-01-01

    A detailed computer model of human erythrocyte metabolism was shown to predict three steady states, two stable and one unstable. The most extreme steady state is characterized by almost zero concentrations of all the phosphorylated intermediates. The "normal" steady state is remarkably robust in the face of large changes in the activity of most of the enzymes of glycolysis and the pentose phosphate pathway: this steady state can be viewed as an attractor towards which the system returns following a metabolic perturbation. Focus is given to three responses of the system: (1) the 'energy charge' that pertains to the concentration of ATP relative to all purine nucleotides; (2) redox power expressed as the ratio of reduced-to-total glutathione and (3) the concentration of 2,3-bisphosphoglycerate, that directly affects the oxygen affinity of haemoglobin thus affecting the main physiological function of the cell. The collapse of the normal steady state in what can be viewed topologically as a catastrophe is posited as one key element of erythrocyte senescence and it is particularly important for erythrocyte destruction in patients with an inborn enzyme deficiency.

  20. 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

  1. The use of erythrocyte fragility to assess xenobiotic cytotoxicity.

    PubMed

    Pagano, Maria; Faggio, Caterina

    2015-08-01

    The erythrocytes of mammals represent a good model to evaluate the cytotoxicity of molecules, organic and inorganic, natural or synthetic, by cellular damage measure. Indeed, before any investigation on the mechanism of action of different molecules, it is important to perform a cytotoxicity assay. Among the different cytotoxicity assays that assess a possible toxicity in the red blood cells is the rate of haemolysis. This essay is based on the evaluation of the alterations of red cell membranes in the presence of an eventual xenobiotic. Red blood cells are the main cells in circulation, and they are responsible for transporting oxygen; in fact, any alterations of this process could be lethal. The plasma membrane of red blood cells is a multi-component structure such as to confer to these cells their characteristic biconcave shape, high flexibility, elasticity and deformability. However, there are clear signs of cellular suffering if there are any alterations to this structure. One method of toxicity assessment is based on measurement of the efflux of haemoglobin from suspended red blood cells. Haemolysis, and therefore the loss of haemoglobin, is the signal stability of the cell membrane of the erythrocytes. In recent years, the discovery of programmed cell death in mammalian red blood cells presented a diversification of the response to injury by these a-nucleated cells. This review shows that mammals' erythrocytes might serve well as a model cell to study on the cellular and molecular mechanisms of many treatments.

  2. Epigenetic Silencing of Plasmodium falciparum Genes Linked to Erythrocyte Invasion

    PubMed Central

    Cortés, Alfred; Carret, Celine; Kaneko, Osamu; Yim Lim, Brian Y. S.; Ivens, Alasdair; Holder, Anthony A

    2007-01-01

    The process of erythrocyte invasion by merozoites of Plasmodium falciparum involves multiple steps, including the formation of a moving junction between parasite and host cell, and it is characterised by the redundancy of many of the receptor–ligand interactions involved. Several parasite proteins that interact with erythrocyte receptors or participate in other steps of invasion are encoded by small subtelomerically located gene families of four to seven members. We report here that members of the eba, rhoph1/clag, acbp, and pfRh multigene families exist in either an active or a silenced state. In the case of two members of the rhoph1/clag family, clag3.1 and clag3.2, expression was mutually exclusive. Silencing was clonally transmitted and occurred in the absence of detectable DNA alterations, suggesting that it is epigenetic. This was demonstrated for eba-140. Our data demonstrate that variant or mutually exclusive expression and epigenetic silencing in Plasmodium are not unique to genes such as var, which encode proteins that are exported to the surface of the erythrocyte, but also occur for genes involved in host cell invasion. Clonal variant expression of invasion-related ligands increases the flexibility of the parasite to adapt to its human host. PMID:17676953

  3. Acid-sensitive outwardly rectifying anion channels in human erythrocytes.

    PubMed

    Kucherenko, Yuliya V; Mörsdorf, Daniel; Lang, Florian

    2009-07-01

    Acid-sensitive outwardly rectifying anion channels (ASOR) have been described in several mammalian cell types. The present whole-cell patch-clamp study elucidated whether those channels are expressed in erythrocytes. To this end whole-cell recordings were made in human erythrocytes from healthy donors treated with low pH and high osmotic pressure. When the pipette solution had a reduced Cl(-) concentration, treatment of the cells with Cl(-)-containing normal and hyperosmotic (addition of sucrose and polyethelene glycol 1000 [PEG-1000] to the Ringer) media with low pH significantly increased the conductance of the cells at positive voltages. Channel activity was highest in the PEG-1000 media (95 and 300 mM PEG-1000, pH 4.5 and 4.3, respectively) where the current-voltage curves demonstrated strong outward rectification and reversed at -40 mV. Substitution of the Cl(-)-containing medium with Cl(-)-free medium resulted in a decrease of the conductance at hyperpolarizing voltages, a shift in reversal potential (to 0 mV) and loss of outward rectification. The chloride currents were inhibited by chloride channels blockers DIDS and NPPB (IC(50) for both was approximately 1 mM) but not with niflumic acid and amiloride. The observations reveal expression of ASOR in erythrocytes.

  4. Biorheological action of Ascaris lumbricoides larvae on human erythrocytes.

    PubMed

    de León, Patricia Ponce; Del Balzo, Gonzalo; Riquelme, Bibiana

    2013-03-01

    Previous studies have shown that A. lumbricoides extracts capture sialic acid (SA) from human red blood cells (RBC). The aim of this work was to study hemorheological alterations in vitro caused by parasite larvae. The biorheological action of three larva concentrates of first and second larval stage on group O erythrocytes was analyzed by incubating the erythrocyte packed together with an equal volume of larvae (treated RBC) and PBS (control RBC). Distribution and parameters of aggregation (digital image analysis), aggregation kinetics (erythroaggregameter), and viscoelasticity (erythrodeformeter) were measured. The digital image analysis showed that all the larvae diminished the isolated cells percentage and increased the size of the formed aggregates. The aggregate formation velocity was lower in the treated than in the control. The deformability index (ID) values of treated RBC did not present variations with respect to those of the control, but a decrease in the erythrocyte elastic modulus (μ(m)) and membrane surface viscosity (η(m)) values was observed, indicating that the larvae not only induced a diminution in the membrane surface viscosity of RBC but also altered the dynamic viscoelasticity of the membrane. Experiments carried out in vitro support the conclusion that the contact between larvae and RBC produces hemorheological alterations.

  5. Microelectrode-based dielectric spectroscopy of glucose effect on erythrocytes.

    PubMed

    Colella, L; Beyer, C; Fröhlich, J; Talary, M; Renaud, P

    2012-06-01

    The dielectric response of biconcave erythrocytes exposed to D-glucose and L-glucose has been investigated using a double array of planar interdigitated microelectrodes on a glass microchip. Erythrocytes are analyzed under physiological conditions suspended in hypo-osmolar balanced solutions containing different glucose concentrations (0-20 mM). The glucose effect on the cellular dielectric properties is evaluated by analyzing the spectra using two different approaches, the equivalent circuit model and a modified model for ellipsoidal particles. The results show that at elevated glucose concentration (15 mM) the membrane capacitance increases by 36%, whereas the cytosol conductivity slightly decreases with a variation of about 15%. On the contrary, no variation has been registered with L-glucose, a biologically inactive enantiomer of D-glucose. The paper discusses the possible mechanism controlling the membrane dielectric response. As the external D-glucose increases, the number of activated glucose transporter in the erythrocyte membrane raises and the transition from sugar-free state to sugar-bounded state induces a change in the dipole moments and in the membrane capacitance.

  6. Regulation of actin filament length in erythrocytes and striated muscle.

    PubMed

    Fowler, V M

    1996-02-01

    Actin filaments polymerize in vitro to lengths which display an exponential distribution, yet in many highly differentiated cells they can be precisely maintained at uniform lengths in elaborate supramolecular structures. Recent results obtained using two classic model systems, the erythrocyte membrane cytoskeleton and the striated muscle sarcomere, reveal surprising similarities and instructive differences in the molecules and mechanisms responsible for determining and maintaining actin filament lengths in these two systems. Tropomodulin caps the slow-growing, pointed filament ends in muscle and in erythrocytes. CapZ caps the fast-growing, barbed filament ends in striated muscle, whereas a newly discovered barbed end capping protein, adducin, may cap the barbed filament ends in erythrocytes. The mechanisms responsible for specifying the characteristic filament lengths in these systems are more elusive and may include strict control of the relative amounts of actin filament capping proteins and side-binding proteins, molecular templates (e.g. tropomyosin and nebulin) and/or verniers (e.g. tropomyosin).

  7. Pyrithione and 8-hydroxyquinolines transport lead across erythrocyte membranes.

    PubMed

    Lind, Stuart E; Park, Jong Sung; Drexler, John W

    2009-09-01

    Acute and chronic lead poisoning remains a significant health problem. Although chelating agents can bind to plasma lead, they cannot cross cell membranes where the total body lead burden resides, and are thus inefficient at reducing the total body lead burden. Recently, calcium and sodium ionophores have been shown to transport lead across cell membranes providing a novel method for reducing total body lead stores. We recently found that clioquinol, an 8-hydroxyquinoline derivative, can act as a zinc ionophore. We postulated that zinc ionophores might also be able to transport lead across biological membranes. To study this, we loaded lead in vitro into human erythrocytes and then studied the ability of zinc ionophores to transport lead into the extracellular space, where it was trapped with a lead chelator. Using inductively coupled plasma mass spectrometry (ICP-MS), we found that several 8-hydroxyquinoline derivatives, as well as the zinc and sodium salts of pyrithione (N-hydroxypyridine-2-thione), reduced erythrocyte lead content. The water-soluble compound, sodium pyrithione, was able to reduce lead in citrated whole blood, without partitioning into the erythrocytes. These results indicate that two classes of zinc ionophores can transport lead across a biological membrane, and they confirm that these ionophores are not cation-specific. Lead ionophores may prove useful in mobilizing lead into the extracellular space, thereby improving the efficacy of chelation therapy, in vivo or ex vivo.

  8. 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

  9. Drug-loaded erythrocytes: on the road toward marketing approval.

    PubMed

    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.

  10. Micronucleated erythrocytes in newborn rats exposed to raltegravir placental transfer.

    PubMed

    Torres-Mendoza, Blanca Miriam; Coronado-Medina, Damharis Elizabeth; Gómez-Meda, Belinda Claudia; Vázquez-Valls, Eduardo; Zamora-Perez, Ana Lourdes; Lemus-Varela, María de Lourdes; Zúñiga-González, Guillermo Moisés

    2014-01-01

    The use of raltegravir in treating HIV/AIDS has been proposed due to its effectiveness in suppressing high loads of HIV RNA in pregnant women, thus preventing infection of the fetus. However, administration of raltegravir during pregnancy produces a compound which is transferred to high concentrations to the offspring. The objective of this study is to evaluate the transplacental genotoxic effect of raltegravir in newborn rats. We evaluated the number of micronucleated erythrocytes (MNE), micronucleated polychromatic erythrocytes (MNPCE), and polychromatic erythrocytes (PCE) in the peripheral blood samples of the offspring of Wistar rats treated 6 days before birth with oral administration of raltegravir. The animals were randomly assigned to five groups as follows: raltegravir at doses of 15, 30, or 60 mg/day, cyclophosphamide 10 mg/kg (positive control), or 0.5 ml of sterile water (negative control). In addition, the effect of these drugs on the weight and height of newborns was assessed. There were no differences in the number of MNE, MNPCE, and PCE, and a slight decrease in the weight and height was observed in the offspring of the rat mothers treated with raltegravir. Genotoxicity studies are required in pregnant women to determine the risk of using raltegravir to the fetuses.

  11. Interactions between peroxiredoxin 2, hemichrome and the erythrocyte membrane.

    PubMed

    Bayer, Simone B; Low, Felicia M; Hampton, Mark B; Winterbourn, Christine C

    2016-12-01

    Peroxiredoxin 2 (Prx2) is an abundant antioxidant protein in erythrocytes that protects against hemolytic anemia resulting from hemoglobin oxidation and Heinz body formation. A small fraction of Prx2 is bound to the cell membrane, but the mechanism and relevance of binding are not clear. We have investigated Prx2 interactions with the erythrocyte membrane and oxidized hemoglobin and whether these interactions are dependent on Prx2 redox state. Membrane binding of Prx2 in erythrocytes decreased when the cells were treated with H2O2, but studies with purified Prx2 and isolated ghosts showed that the interaction was independent of Prx2 redox state. Hemoglobin oxidation leads to the formation of hemichrome, a denatured form of the protein that binds to Band3 protein in the cell membrane as part of the senescence process and is a precursor of Heinz bodies. Hemichrome competed with Prx2 and decreased Prx2 binding to the membrane, potentially explaining the decreased binding in oxidant-exposed cells. The increased membrane binding of Prx2 seen with increasing intracellular calcium was less sensitive to H2O2 or hemichrome, suggesting an alternative mode of binding. Prx2 was also shown to exhibit chaperone-like activity by retarding the precipitation of pre-formed hemichrome. Our results suggest that Prx2, by restricting membrane binding of hemichrome, could impede Band3 clustering and exposure of senescence antigens. This mechanism, plus the observed chaperone activity for oxidized hemoglobin, may help protect against hemolytic anemia.

  12. Dielectric relaxations on erythrocyte membrane as revealed by spectrin denaturation.

    PubMed

    Ivanov, I T; Paarvanova, B

    2016-08-01

    We studied the effect of spectrin denaturation at 49.5°C (TA) on the dielectric relaxations and related changes in the complex impedance, Z*, complex capacitance, C*, and dielectric loss curve of suspensions containing human erythrocytes, erythrocyte ghost membranes (EMs) and Triton-X-100 residues of EMs. The loss curve prior to, minus the loss curve after TA, resulted in a bell-shaped peak at 1.5MHz. The changes in the real and imaginary components of Z* and C* at TA, i.e., ΔZre, ΔZim, ΔCre and ΔCim, calculated in the same way, strongly varied with frequency. Between 1.0 and 12MHz the -ΔZim vs ΔZre, and ΔCim vs ΔCre plots depicted semicircles with critical frequencies, fcr, at 2.5MHz expressing recently reported relaxation of spectrin dipoles. Between 0.02 and 1.0MHz the -ΔZim vs ΔZre plot exhibited another relaxation whose fcr mirrored that of beta relaxation. This relaxation was absent on Triton-X-shells, while on erythrocytes and EMs it was inhibited by selective dissociation of either attachment sites between spectrin and bilayer. Considering above findings and inaccessibility of cytosole to outside field at such frequencies, the latter relaxation was assumed originating from a piezoelectric effect on the highly deformable spectrin filaments.

  13. 7Li NMR study of normal human erythrocytes

    NASA Astrophysics Data System (ADS)

    Pettegrew, J. W.; Post, J. F. M.; Panchalingam, K.; Withers, G.; Woessner, D. E.

    The biological action of lithium is of great interest because of the therapeutic efficacy of the cation in manic-depressive illness. To investigate possible molecular interactions of lithium, 7Li NMR studies were conducted on normal human erythrocytes which had been incubated with lithium chloride. The uptake of lithium ions was followed by 7Li NMR, using a dysprosium, tripolyphosphate shift reagent. Lithium uptake followed single-exponential kinetics with a time constant of 14.7 h. The intracellular lithium relaxation times were T 1 ⋍ 5 s and T 2 ⋍ 0.15 s, which implies a lengthening of the lithium correlation time. It was found that lithium does not interact significantly with hemoglobin, the erythrocyte membrane, or artificial phospholipid membranes. Based on measurements of lithium T1 and T2 in concentrated agar gels, the large difference between T1 and T2 for intracellular lithium ions may be due to diffusion of the hydrated lithium ion through heterogeneous electrostatic field gradients created by the erythrocyte membrane-associated cytoskeletal network. Lithium binding to the membrane-associated cytoskeleton, however, cannot be ruled out. Because of the large differences between T1 and T2 of intracellular lithium ions, 1Li NMR may be a sensitive and promising noninvasive method to probe the intracellular environment.

  14. Micronucleated Erythrocytes in Newborn Rats Exposed to Raltegravir Placental Transfer

    PubMed Central

    Torres-Mendoza, Blanca Miriam; Coronado-Medina, Damharis Elizabeth; Vázquez-Valls, Eduardo; Zamora-Perez, Ana Lourdes; Lemus-Varela, María de Lourdes

    2014-01-01

    The use of raltegravir in treating HIV/AIDS has been proposed due to its effectiveness in suppressing high loads of HIV RNA in pregnant women, thus preventing infection of the fetus. However, administration of raltegravir during pregnancy produces a compound which is transferred to high concentrations to the offspring. The objective of this study is to evaluate the transplacental genotoxic effect of raltegravir in newborn rats. We evaluated the number of micronucleated erythrocytes (MNE), micronucleated polychromatic erythrocytes (MNPCE), and polychromatic erythrocytes (PCE) in the peripheral blood samples of the offspring of Wistar rats treated 6 days before birth with oral administration of raltegravir. The animals were randomly assigned to five groups as follows: raltegravir at doses of 15, 30, or 60 mg/day, cyclophosphamide 10 mg/kg (positive control), or 0.5 ml of sterile water (negative control). In addition, the effect of these drugs on the weight and height of newborns was assessed. There were no differences in the number of MNE, MNPCE, and PCE, and a slight decrease in the weight and height was observed in the offspring of the rat mothers treated with raltegravir. Genotoxicity studies are required in pregnant women to determine the risk of using raltegravir to the fetuses. PMID:24977162

  15. Disorders in the Morphology and Nanostructure of Erythrocyte Membranes after Long-term Storage of Erythrocyte Suspension: Atomic Force Microscopy Study.

    PubMed

    Moroz, V V; Chernysh, A M; Kozlova, E K; Sergunova, V A; Gudkova, O E; Khoroshilov, S E; Onufrievich, A D; Kostin, A I

    2015-07-01

    Disorders in the erythrocyte morphology and structure of their membranes during long-term storage of erythrocyte suspension (30 days at 4°C) were studied by atomic force microscopy. The morphology and nanostructure of erythrocyte membranes, biochemical parameters, ion exchange parameters, and hemoglobin spectra were recorded. The transformation of erythrocyte morphology and destruction of their membranes were observed throughout the storage period. Irreversible forms of spheroechinocytes and their fragments formed by the end of storage. The concentrations of potassium ions and lactate in solution of the blood preservatiive increased, while pH value decreased. Hemolysis detected by the erythrocyte "leakage" effect was observed starting from days 16-23 of storage.

  16. A membrane-impermeant, cleavable cross-linker. Dimers of human erythrocyte band 3 subunits cross-linked at the extracytoplasmic membrane face.

    PubMed

    Staros, J V; Morgan, D G; Appling, D R

    1981-06-10

    We have synthesized diisethionyl-3,3'-dithiobispropionimidate (DIDIT), a new membrane-impermeant, cleavable protein cross-linking reagent designed for probing protein organization at one face of a membrane. Rabbit muscle aldolase were reacted in solution with DIDIT and the products were electrophoresed in sodium dodecyl sulfate-polyacrylamide gels. When electrophoresed under nonreducing conditions, the gels contain bands corresponding to oligomers of aldolase, while pretreatment with dithiothreitol to cleave the cross-link prior to electrophoresis results in gels containing primarily the band corresponding to aldolase monomer. These experiments demonstrate that DIDIT is a cleavable protein cross-linker. Reaction of isolated human erythrocyte membranes with DIDIT leads to extensive cross-linking of spectrin, band 3, and band 6, and residual hemoglobin, consistent with results previously obtained with permeant cross-linkers. In contrast, when intact human erythrocytes are cross-linked with DIDIT, hemoglobin and the cytoplasmic face membrane proteins are not cross-linked, but band 3, which is accessible at the extracytoplasmic face of the membrane, is cross-linked to dimers.

  17. Erythrocyte caspase-3 activation and oxidative imbalance in erythrocytes and in plasma of type 2 diabetic patients.

    PubMed

    Maellaro, Emilia; Leoncini, Silvia; Moretti, Daniele; Del Bello, Barbara; Tanganelli, Italo; De Felice, Claudio; Ciccoli, Lucia

    2013-08-01

    An increased oxidative stress and a decreased life span of erythrocytes (RBCs) are reported in patients with diabetes. Aim of this study was to assess in RBCs from patients with type 2 diabetes whether downstream effector mechanisms of apoptosis, such as activation of caspase-3, is operative, and whether an iron-related oxidative imbalance, occurring inside RBCs and in plasma, could be involved in caspase-3 activation. In 26 patients with type 2 diabetes and in 12 healthy subjects, oxidative stress was evaluated by means of different markers; non-protein-bound iron, methemoglobin and glutathione were determined in RBCs, and non-protein-bound iron was also determined in plasma. Erythrocyte caspase-3 activation was evaluated by an immunosorbent enzyme assay. Arterial hypertension, demographic and standard biochemical data were also evaluated. The results show, for the first time, that type 2 diabetic RBCs put into motion caspase-3 activation, which is significantly higher than in control RBCs. Such an effector mechanism of "eryptosis" was positively correlated to blood glucose levels and to the increased plasma NPBI level. Caspase-3 activation was also positively correlated to occurrence of arterial hypertension. The results suggest that an extracellular oxidative milieu can be responsible for erythrocyte caspase-3 activation in patients with type 2 diabetes. In turn, caspase-3 activation can be envisaged as a novel mechanism which, by impairing the maintenance of erythrocyte shape and function, might contribute to the shortened life span of RBCs from patients with type 2 diabetes and to hemorheological disorders observed in these patients.

  18. Erythrocyte: A systems model of the control of aggregation and deformability.

    PubMed

    Bazanovas, Antonina N; Evstifeev, Aleksandr I; Khaiboullina, Svetlana F; Sadreev, Ildar I; Skorinkin, Andrey I; Kotov, Nikolay V

    2015-05-01

    Human erythrocytes are highly specialized enucleate cells that are involved in providing efficient gas transport. Erythrocytes have been extensively studied both experimentally and by mathematical modeling in recent years. However, understanding of how aggregation and deformability are regulated is limited. These properties of the erythrocyte are essential for the physiological functioning of the cell. In this work, we propose a novel mathematical model of the molecular system that controls the aggregation and deformability of the erythrocyte. This model is based on the experimental results of previously published studies. Our model suggests fundamentally new mechanisms that regulate aggregation and deformability in a latch-like manner. The results of this work could be used as a general explanation of how the erythrocytes regulate their aggregation and deformability, and are essential in understanding erythrocyte disorders and aging.

  19. Isolation of proteins related to the Rh polypeptides from nonhuman erythrocytes.

    PubMed Central

    Saboori, A M; Denker, B M; Agre, P

    1989-01-01

    It is thought that the Rh antigens may be important in maintaining normal erythrocyte membrane integrity. Despite their name, Rh antigens are serologically present only on human erythrocytes. Rh structural polymorphisms are known to reside within a family of nonglycosylated Mr 32,000 integral membrane proteins that can be purified by hydroxylapatite chromatography. Mr 32,000 integral membrane proteins were purified similarly from erythrocyte membrane vesicles prepared from rhesus monkeys, cows, cats, and rats, but could not be purified from human Rhmod erythrocytes, a rare syndrome lacking Rh antigens. The purified Mr 32,000 polypeptides were labeled with 125I, digested with chymotrypsin, and found to be 30-60% identical to human Rh polypeptides when compared by two-dimensional iodopeptide mapping. The physiologic function of the Rh polypeptides remains to be identified; however, the existence of related proteins in nonhuman erythrocytes supports the concept that the Rh polypeptides are erythrocyte membrane components of fundamental significance. Images PMID:2492035

  20. [Raman spectra of different kinds of thalassemia erythrocytes with the effect of pH].

    PubMed

    Wu, Zheng-Jie; Wang, Cheng; Lin, Zheng-Chun

    2013-04-01

    Thalassemia is a kind of blood diseases which has high morbidity and large influence. Previous methods for diagnosis are all very cumbersome and time consuming. By comparing Raman spectra of different kinds of thalassemia and normal erythrocytes at acid or alkaline pH, it was found that beta-thalassemia and alpha-thalassemia erythrocytes have dissimilar Raman spectra in the acidic environment, such as the Raman spectra of beta-thalassemia erythrocytes showed higher intensity at the characteristic bands assigned to oxyhemoglobin, and the characteristic bands assigned to deoxyhemoglobin were even completely replaced; beta-thalassemia erythrocytes membrane has a smaller chain interaction between the transverse order parameters than normal erythrocytes, and the S(lat) values are different for different stages of anemia, while the S(lat) values are similar between alpha-thalassemia and normal erythrocytes, indicating that based on the effect of pH it is possible to diagnose thalassemia more quickly by using Raman spectra.